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Purpose: Hypoxia-inducible factor 1α (HIF-1α) activity is one of the major players in hypoxia-mediated glioma progression and resistance to therapies, and therefore the focus of this study was the evaluation of HIF-1α modulation in relation to tumour response with the purpose of identifying imaging biomarkers able to document tumour response to treatment in a murine glioma model. Methods: U251-HRE-mCherry cells expressing Luciferase under the control of a hypoxia responsive element (HRE) and mCherry under the control of a constitutive promoter were used to assess HIF-1α activity and cell survival after treatment, both in vitro and in vivo, by optical, MRI and positron emission tomography imaging. Results: This cell model can be used to monitor HIF-1α activity after treatment with different drugs modulating transduction pathways involved in its regulation. After temozolomide (TMZ) treatment, HIF-1α activity is early reduced, preceding cell cytotoxicity. Optical imaging allowed monitoring of this process in vivo, and carbonic anhydrase IX (CAIX) expression was identified as a translatable non-invasive biomarker with potential clinical significance. A preliminary in vitro evaluation showed that reduction of HIF-1α activity after TMZ treatment was comparable to the effect of an Hsp90 inhibitor, opening the way for further elucidation of its mechanism of action. Conclusion: The results of this study suggest that the U251-HRE-mCherry cell model can be used for the monitoring of HIF-1α activity through luciferase and CAIX expression. These cells can become a useful tool for the assessment and improvement of new targeted tracers for potential theranostic procedures. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to characterize a cell-based model for the molecular study of hypoxia-inducible factor (HIF)-1α activity, in the context of hypoxia, by means of different imaging techniques. Procedures: Engineered U251-HRE glioma cells were used to analyze the molecular mechanisms underlying HIF-1α activity in vitro in relation to luciferase expression. The same cells were orthotopically implanted in mice to evaluate tumor progression and hypoxia induction by bioluminescence imaging, fluorescence imaging, positron emission tomography (PET), and magnetic resonance imaging (MRI). Results: In vitro analyses highlighted the relationship between HIF-1α and luciferase activity in hypoxic conditions and after pharmacological treatments in U251-HRE cells. Through in vivo studies, it was possible to assess hypoxia establishment in relation to tumor growth by optical imaging, PET and MRI. Conclusions: The findings of this study indicate that the U251-HRE orthotopic murine model can be used to reliably evaluate processes modulating HIF-1α activity, using both molecular and preclinical non-invasive imaging techniques. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate circadian variations of blood and milk lead toxicokinetics in dairy cows in summer. Twenty lactating Holstein animals were randomly assigned to four treatments corresponding to different hours after onset of light (HALO): 2, 8, 14, and 20. Cows received a single intravenous administration of 2.5 mg/kg lead as lead acetate. Blood and milk samples were taken and analyzed by atomic absorption spectrophotometry. For each toxicokinetic parameter, a one-way analysis of variance (ANOVA) was performed to outline the existence of daily variations. Significant blood differences as a function of HALO were found for the hybrid constant of distribution (α), hybrid constant of elimination (β), elimination half-life ( $${\text{t}}_{{{\text{1/2 $ \beta $ }}}} $$), area under the curve (AUC), volume of distribution at steady state (Vss) and clearance (ClB) ( p<0.05). Half-life of elimination presented two peaks at 2 and 14 HALO. Milk data showed significant differences for maximum concentration and AUC ( p<0.05). The ratio AUCmilk/AUCblood was utilized to estimate penetration of lead in milk. It differed significantly throughout the day ( p<0.05). Milk data for the significant parameters could be fitted to circadian rhythms. No circadian rhythms were detected in blood parameters or in the ratio AUCmilk/AUCblood. [ABSTRACT FROM AUTHOR]

Thirty-two multiparous Holando argentino cows in mid lactation were randomly assigned to two treatments: control or HFF (hydrogenated fish fat) at Rafaela, 31° 11’ South, during summer 1997/1998, to evaluate the effect of using HFF as a supplement under grazing conditions. Animals in both treatments grazed an alfalfa pasture, and were confined from 1000 hours to 1700 hours daily in a shaded pen where water was provided ad libitum. During each milking, animals in the control group received 3.73 kg dry matter per cow each day (DM cow–1 day–1) concentrate (15% crude protein; 8.69 MJ energy for location/kg DM). Cows in the HFF group received 3.25 kg DM cow–1 day–1 concentrate, plus 0.200 kg DM cow–1 day–1 HFF. Both diets presented similar energy, protein and neutral detergent fibre contents. The trial was performed during a strong ”El Niño” event, which resulted in a total rainfall of 396.3 mm (80% higher than normal). The mean temperature was 23.7 (SD 3.2)°C and the mean temperature humidity index was 72.9 (SD 4.96). Production data were analysed using a completely randomised design with analysis of covariance. Supplementation with HFF produced an increase in daily milk production (26.4 (SD 2.46) l/cow compared to 23.9 (SD 2.68) l/cow for the controls; P<0.05). Milk fat production was higher for HFF ( P<0.05): 941 (SD 96) g cow–1 day–1 as compared to controls, which yielded 846 (SD 95) g cow–1 day–1. Milk protein yields also differed significantly ( P<0.05), the respective values for HFF and controls being 795 (SD 72) g cow–1 day–1 and 715 (SD 83) g cow–1 day–1. It was concluded that hydrogenated fish fat could be a good ingredient to sustain high yields and elevated maintenance requirements in a grazing system during hot conditions. [ABSTRACT FROM AUTHOR]

Two tree shades (TS1 and TS2) and an artificial shade structure (AS) were evaluated using black globe temperatures (BGTs) to assess their effectiveness in reducing heat load. The artificial structure consisted of a black woven polypropylene cloth providing 80% shade, mounted on 2.5-m-high eucalyptus posts. The work was carried out at Rafaela Experimental Station, Argentina, during the summer (January and February) 1994. BGTs and floor temperatures were measured in concrete floor holding pens with and without artifical shade. The results showed no difference between TS1, TS2 and AS, their average BGTs being 30.2 (SD 0.58), 29.0 (SD 0.70) and 30.2 (SD 0.74)°C, respectively. BGTs under all three shades were significantly lower ( P<0.01) than the average BGT recorded outside: 35.5 (SD 1.12)°C. Average BGTs in holding pens were 32.4 (SD 1.38) and 39.9 (SD 1.91)°C for shaded and non-shaded areas ( P<0.01). The corresponding average floor temperatures were 27.8 (SD 0.68)°C and 47.7 (SD 2.13)°C ( P<0.01). To assess the effects of shade on animal well-being, afternoon rectal temperatures (RT) and respiratory rate (respirations per minute, RR) of lactating cows were recorded twice a week. Rectal temperatures were significantly higher for non-shaded cows ( P<0.01), mean RT being 40.1 (SD 0.65)°C vs 39.3 (SD 0.42)°C for the shaded animals. Corresponding RRs were 78.9 (SD 18.0) and 60.7 (SD 10.6) ( P<0.05). It was concluded that: (1) tree and artificial shades produced similar effects, (2) shading the holding pen with an 80% shading cloth was effective in reducing heat load and floor temperatures, and (3) access to shade in our pasture-based system improved animal well-being. [ABSTRACT FROM AUTHOR]