Single Sc Research Articles

Effects of amifostine on endometriosis, comparison with N-acetyl cysteine, and leuprolide as a new treatment alternative: a randomized controlled trial

To assess the effects of amifostine, N-acetyl cysteine (NAC), and leuprolide as a scavenger in a rat endometriosis model. This is a prospective randomized animal study. Setting The Animal Laboratory of Medical University. Animals 40 rats were used for transplantation of an autologous fragment of endometrial tissue onto the inner surface of the abdominal wall. After allowing 3 weeks for growth, laparotomies were performed to check the implants. Then animals were randomized into four groups: Group I amifostine (200 mg/day loading dose after 20 mg/kg/day, p.o.); Group II NAC (200 mg/day, p.o.); Group III leuprolide acetate 1 mg/kg single dose, sc; and Group IV (controls) no medication. Three weeks later, implants were evaluated morphologically. Serum and peritoneal TNF-alpha levels were evaluated. The transmission electron microscopic examination of the peritoneal samples and ovaries was also performed. Leuprolide acetate, amifostine and NAC caused significant decreases in the mean implant areas and significant decreases in serum and peritoneal TNF-alpha levels. On comparing all groups, these reductions were higher in Group II. According to the transmission electron microscopic findings, leuprolide seems to be protecting normal structure of peritoneum best when compared to the other groups. Amifostine, NAC and leuprolide caused regression of endometriosis in this experimental rat model by a yet unsettled mechanism.

Abstract LB-185: Inhibitory effects of black raspberry consumption on prostate carcinogenesis in rats.

Abstract Black raspberries are being developed to chemoprevent GI tract cancers in humans and they have been shown to prevent breast cancer development in a rat model. Therefore, it is important to determine whether they can also prevent other major human cancers such as prostate cancer. We induced prostate cancer in Wistar HsdCpb:WU rats by treatment with Flutamide (20 mg/kg BW by gavage daily for 20 days), a single SC injection with testosterone propionate (100 mg/kg) on day 21, and single IP injection of N-methyl-N-nitrosourea (MNU; 30 mg/kg) 52-56 hrs later. Two weeks later, rats received two SC Silastic tubing implants (ID 0.078 in; OD 0.125 in.) containing crystalline testosterone tightly packed over 3 cm length. One week later, rats were started on AIN-93M diet (control) or isoenergetic AIN-93M diets containing 5% or 10% lyophilized black raspberry powder (exchanged for starch). The powder was from a single batch of freshly harvested black raspberries (BerriProducts, Corvallis, OR), stored at -20oC until mixed into the diet, and provided fresh to the rats thrice weekly. The treatment did not affect body weight gain or survival (mean 49-52 wks). The study was terminated 58 weeks after MNU and at necropsy prostate complexes and all tumor-like lesions were collected for histology. Prostate cancer incidence was determined by enumerating grossly observed tumors and histopathologic evaluation using step sections. Overall incidence of large prostate tumors, histologically confirmed as adenocarcinomas, was reduced from 16/31 rats in the control group (52%) to 14/30 (46%) and 8/30 (27%) rats in the low and high dose black raspberry groups, resp., which trend was significant (p = 0.049; X2 test). The difference in incidence between control and high dose groups was borderline significant (p = 0.067, 2-sided; p = 0.41, 1-sided Fisher exact test). In subgroups of 4-5 rats, the prostate was collected after 3 weeks of feeding the diets and the concentrations of the black raspberry anthocyanin metabolite protocatechuic acid (PCA) were measured in the lateral prostate using LC-MS/MS. Mean (± SD) prostatic concentrations of PCA (ng/g wet tissue) were 0.5 ± 0.0 (detection limit), 3.64 ± 1.35 (p<0.05), and 4.61 ± 1.92 (p<0.01) in the control, low, and high dose groups, resp. (overall p = 0.009; Kruskall-Wallis test). Preliminary results of in vitro studies with black raspberry constituents indicated that ellagic acid, which is more stable than anthocyanins, reduced growth of human LAPC-4 prostate cancer cells by ≥ 25% at concentrations of 10 μg/ml and higher. These results indicate that black raspberry consumption (1) can inhibit prostate cancer induction in rats, (2) results in significant amounts of an anthocyanin metabolite in the rat prostate, and (3) ellagic acid, a major bioactive agents in black raspberries, can inhibit in vitro growth of human prostate cancer cells. (Supported by NIH Grant CA152879). Citation Format: Maarten C. Bosland, Michael J. Schlicht, Jillian Eskra, Zhongfa Liu. Inhibitory effects of black raspberry consumption on prostate carcinogenesis in rats. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-185. doi:10.1158/1538-7445.AM2013-LB-185

Abstract 679: SGI-110 and entinostat therapy reduces lung tumor burden and reprograms the epigenome .

Abstract The DNA methyltransferase inhibitors (DNMTi) Vidaza (5-azacytidine) and Dacogen (5-aza-2′-deoxycytidine, decitabine) are approved by the FDA for chemotherapy against myelodysplastic syndrome, a heterogeneous bone marrow disorder. Vidaza in combination with the HDAC inhibitor entinostat has shown some promise in treating lung cancer and this has been replicated in our orthotopic lung cancer model. However, the effectiveness of these DNMTi against solid tumors is likely impacted in part, by their stability and rapid inactivation by cytidine deaminase in the liver. These studies were initiated to test the efficacy of SGI-110, a prodrug for decitabine. SGI-110, a dinucleotide consisting of 5-aza-2′-deoxycytidine followed by a deoxyguanosine, has been shown to be effective in vitro as a DNA methylation inhibitor and less prone to deamination by cytidine deaminase. In this study, we used our in vivo lung cancer model to explore the potential therapeutic effect of SGI-110 alone and in combination with entinostat. In vivo plasma concentrations and pharmacokinetic properties of SGI-110 and its metabolite decitabine were evaluated in nude rats after a single SC dose of SGI-110. Upon SC administration, SGI-110 is rapidly converted to decitabine. Dose proportional pharmacokinetics showed no significant difference in accumulation of SGI-110 or decitabine, while the half life of decitabine was increased to 4hrs. SGI-110 alone or in combination with entinostat significantly reduced tumor burden by 56% of Calu-6, a K-ras/p53 mutant lung adenocarcinoma cell line engrafted orthotopically in nude rats. On a molecular level, global changes in gene profiles were associated with a reduction in tumor burden. Microarray analysis revealed that a 212 and 592 genes were differentially expressed in SGI-110 alone or with combination therapy. Highly significant gene expression changes were seen in key cancer regulatory pathways. In addition, SGI-110 treatment activated p21 and apoptotic genes BIK and BAK in these tumors. SGI-110 caused widespread epigenetic reprogramming that induced demethylation of an average of 291 genes; in particular 208 genes were polycomb target genes. The polycomb targets were enriched for transcription factors. These preclinical in vivo findings demonstrate that SGI-110 alone shows strong demethylating activity and when combined with entinostat more effectively retards tumor growth. (Supported by Astex Pharmaceutical and SU2C) Citation Format: Carmen S. Tellez, Marcie J. Grimes, Maria A. Picchi, Yushi Liu, Tom H. March, Aram Oganesian, Pietro Taverna, Steven A. Belinsky. SGI-110 and entinostat therapy reduces lung tumor burden and reprograms the epigenome . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 679. doi:10.1158/1538-7445.AM2013-679

Pharmacokinetics, Tissue Distribution and Excretion of Recombinant Human Parathyroid Hormone 1–84 in Animals

To study the plasma pharmacokinetics and accumulation of the recombinant human parathyroid hormone (rhPTH) (1-84) in rhesus monkeys, and the tissue distribution and excretion profiles of (125)I-rhPTH (1-84) in rats. The concentration of rhPTH (1-84) in plasma samples were determined by an enzyme immunoassay (EIA) method after subcutaneous and intravenous bolus injection. The tissue distribution and urinary, fecal and biliary excretion patterns of (125)I-rhPTH (1-84) were investigated by trichloroacetic acid (TCA) precipitation method. Following subcutaneous (sc) administration rhPTH (1-84) in rhesus monkeys, rhPTH (1-84) exhibited rapid absorption and elimination and had no accumulated tendency after successive sc administration. Following sc administration (125)I-rhPTH (1-84) in rats, the TCA-precipitated radioactivity was widely distributed and rapidly diminished in most tissues. Approximately 83.9 and 6.8 % of the total radioactivity was recovered in urine and feces by 72 h postdosing, respectively; whereas 4.1 % excreted into bile up to 24 h postdosing. The pharmacokinetics of rhPTH (1-84) complied with linear kinetics within the examined dose range following a single sc administration had no accumulated tendency following multiple sc administration in rhesus monkeys. The accumulation of (125)I-rhPTH (1-84) in tissues/organs examined, appeared to be low in rats. The major elimination route was by urinary excretion.

Evaluation of a single subcutaneous infusion of carboplatin as adjuvant chemotherapy for dogs with osteosarcoma: 17 cases (2006–2010)

To evaluate adverse effects and survival times in dogs with osteosarcoma that received a single SC infusion of carboplatin as adjunctive chemotherapeutic treatment following limb amputation or limb-sparing surgery. Retrospective case series. 17 client-owned dogs with spontaneously occurring osteosarcoma. Medical records of dogs that underwent limb amputation or limb-sparing surgery followed by a single continuous SC infusion of carboplatin (total dose, 300 mg/m(2) infused over 3, 5, or 7 days) were evaluated. Signalment, tumor location, type of surgery (amputation or limb-sparing), duration of carboplatin infusion, results of hematologic and serum biochemical analyses, and adverse effects were recorded. Kaplan-Meier survival analysis was performed. Median survival time for all dogs was 365 days. Nine dogs had adverse bone marrow-related (hematologic) effects, 1 had adverse gastrointestinal effects, and 7 had infections at the surgical site. No significant differences were detected in survival times of dogs grouped according to tumor location, type of surgery, duration of carboplatin infusion, or development of postoperative infection. Median survival time and adverse effects in dogs with osteosarcoma that received a single SC infusion of carboplatin over a 3-, 5-, or 7-day period as adjunctive treatment following limb amputation or limb-sparing surgery were comparable to those of previously reported chemotherapy protocols requiring IV drug administration over several weeks. Further investigation is needed to evaluate the efficacy of this protocol as adjunctive treatment for osteosarcoma and other tumors in dogs.

Formation of Metal Oxyfluorides from Specific Metal Reactions with Oxygen Difluoride: Infrared Spectroscopic and Theoretical Investigations of the OScF2Radical and OScF with Terminal Single and Triple ScO Bonds

The scandium oxydifluoride free radical, OScF(2), is produced by the spontaneous, specific reaction of laser ablated Sc atoms with OF(2) in solid argon and characterized by using matrix infrared spectroscopy and theoretical calculations. The OScF(2) molecule is predicted to have C(2v) symmetry and a (2)B(2) ground state with an unpaired electron located primarily on the terminal oxygen atom, which makes it a scandium difluoride molecule coordinated by a neutral oxygen atom radical in forming the Sc-O single bond. The closed shell singlet OScF molecule with an obtuse bent geometry has a much shorter Sc-O bond of 1.682 Å than that of the OScF(2) radical (1.938 Å) on the basis of B3LYP calculations. The Sc-O bond in OScF consists of two covalent bonds and a dative bond in which the oxygen 2p(π) lone pair donates electron density into an empty Sc 3d orbital thus forming a triple oxo bond. Density functional calculations suggest it is highly exothermic for fluorine transfer from OF(2) to scandium, which favors the formation of the OScF(2) radical species as well as the OScF molecule after fluorine loss.

Comparison of the efficacy of amoxicillin-clavulanic acid, cefovecin, and doxycycline in the treatment of upper respiratory tract disease in cats housed in an animal shelter

To compare efficacy of amoxicillin-clavulanic acid, cefovecin, and doxycycline in shelter-housed cats with clinical signs of upper respiratory tract disease (URTD). Randomized prospective clinical trial. Animals-48 cats with URTD. Conjunctival and nasal swab specimens were obtained for culture and susceptibility testing, and cats were randomly assigned to 3 treatment groups (16 cats/group) on day 1: amoxicillin-clavulanic acid (12.5 mg/kg [5.68 mg/lb], PO, q 12 h, for 14 days), cefovecin (8.0 mg/kg [3.64 mg/lb], SC, once), or doxycycline (10.0 mg/kg [4.55 mg/lb], PO, q 24 h, for 14 days). Oculonasal discharge, sneezing, coughing, dyspnea, demeanor, and food intake were scored twice daily for 14 days (scale, 0 [subjectively normal] to 3 [markedly abnormal]). The most common bacterial isolates were Mycoplasma spp (n = 22) and Bordetella bronchiseptica (9). Cats treated with amoxicillin-clavulanic acid or doxycycline had significantly increased body weight by day 14. Cats that received doxycycline had significantly lower overall oculonasal discharge scores than those treated with amoxicillin-clavulanic acid or cefovecin. Cats treated with amoxicillin-clavulanic acid or doxycycline had significantly lower overall sneezing scores than those that received cefovecin. Cats that received amoxicillin-clavulanic acid had significantly decreased demeanor and food intake scores on day 2, whereas this was detected later in other groups (demeanor score on days 5 and 7 and food intake score on days 10 and 11 in the cefovecin and doxycycline groups, respectively). Oral administration of amoxicillin-clavulanic acid or doxycycline appeared to be more effective than a single SC injection of cefovecin in treating cats with clinical signs of URTD.

Antagonistic activity of acolbifene, fulvestrant, tamoxifen, and raloxifene on cancer-associated genes in the mouse mammary gland.

10583 Background: The efficacy and exceptionally good tolerance of estrogen blockade in the treatment of breast cancer is well recognized. Acolbifene (ACOL) is a novel and unique SERM completely free of estrogen-like activity in both the mammary gland and uterus. To better understand the specificity of ACOL, we have investigated its effect on the expression of a set of genes modulated by estradiol (E2) in the mouse mammary gland. Methods: ACOL, tamoxifen (TAM), raloxifene (RALOX) and fulvestrant (FULV) were administered (0.01 mg/mouse; sc) to ovariectomized (OVX) mice or to OVX mice simultaneously treated with E2 (0.05 µg/mouse; single sc injection). Microarray screening followed by Q_RTPCR was used to identify a reproducible set of E2 responsive genes. Results: From 128 genes significantly modulated by E2, 108 genes were up-regulated and 20 were down-regulated. Forty-nine of these genes were associated with tumorigenesis while 22 are known to be associated with breast cancer. This set of 49 genes were used to determine the specificity of ACOL compared to another pure antiestrogen in the mammary gland and uterus, namely FULV, as well as to the mixed estrogen antagonists/agonists TAM and RALOX, in their ability to block the effect of E2. Efficacy of reversal of the effect of E2 was 94%, 63%, 45% and 90% for ACOL, FULV, TAM and RALOX, respectively. The overlap between all treatments was 30.6% (15/49). ACOL reversed the effect of E2 on 42 of the 49 (85.7%) cancer-related genes. Between the genes up-regulated by E2 and reversed by ACOL, seven are considered as prognostic markers in breast cancer, namely Fgfr3, Fos12, Junb, Jdp2, Gdf15, Greb1 and Tgm2. On the other hand, two genes down‑regulated by E2, namely Foxa1 and Fgfr2, were restored by ACOL. Conclusions: Taken together, these data offer new information for a better understanding of the previously demonstrated potent tumoricidal action of ACOL in human breast cancer xenografts. The data also suggest, under the tested conditions, superiority of ACOL over TAM and the other compounds to reverse the effect of E2 on specific gene expression, thus supporting the interest of this new 3rd generation SERM for the hormonal therapy and prevention of breast cancer.

Pharmacokinetic behavior of marbofloxacin after intravenous, subcutaneous and intramuscular administrations in llamas (Lama glama)

The purpose of this study was to determine the pharmacokinetic profile of marbofloxacin in llamas following single IV, IM and SC administration of 5mg/kg bw. To assess whether these routes could be an option for the administration of marbofloxacin in this specie and estimate efficacy predictors (PK/PD) from bibliographic MIC90 values. The principal pharmacokinetic parameters were Vss=0.72±0.22L/kg, Cl=0.09±0.03L/kgh (for IV administration), Cmax=7.43±1.28μg/mL and 6.94±2.19μg/mL for IM and SC administration, respectively; t1/2λ=9.16±1.08h, 8.47±0.31h and 6.26±0.87h and MRT=7.30±1.07h, 8.21±1.57h and 8.27±0.96h for IV, IM and SC administrations, respectively. The values obtained for the pharmacokinetic parameters were not significantly different among routes of administration except Cmax, Vz and t1/2λ. The values obtained from the PK/PD indices were Cmax/MIC90 (12.4–32.3) and AUC0–24/MIC90 (105.7–178.52h); taking into account these values, it was concluded that a IM or SC dose of 5mg/(kgday) of marbofloxacin would be adequate in treating infections caused by susceptible bacteria with MIC<0.44μg/mL.

Oral Octreotide Absorption in Human Subjects: Comparable Pharmacokinetics to Parenteral Octreotide and Effective Growth Hormone Suppression

Oral administration of a novel octreotide formulation enabled its absorption to the systemic circulation, exhibiting blood concentrations comparable to those observed with injected octreotide and maintaining its biological activity. The aim of the study was to determine oral octreotide absorption and effects on pituitary GH secretion compared to sc octreotide injection. Four single-dose studies were conducted in 75 healthy volunteers. Oral doses of 3, 10, or 20 mg octreotide and a single sc injection of 100 μg octreotide were administered. We measured the pharmacokinetic profile of orally administrated octreotide and the effect of octreotide on basal and stimulated GH secretion. Both oral and sc treatments were well tolerated. Oral octreotide absorption to the circulation was apparent within 1 h after dose administration. Escalating oral octreotide doses resulted in dose-dependent increased plasma octreotide concentrations, with an observed rate of plasma decay similar to parenteral administration. Both 20 mg oral octreotide and injection of 0.1 mg sc octreotide resulted in equivalent pharmacokinetic parameters [mean peak plasma concentration, 3.77 ± 0.25 vs. 3.97 ± 0.19 ng/ml; mean area under the curve, 16.2 ± 1.25 vs. 12.1 ± 0.45 h × ng/ml); and median time ≥ 0.5 ng/ml, 7.67 vs. 5.88 h, respectively). A single dose of 20 mg oral octreotide resulted in basal (P < 0.05) and GHRH-stimulated (P < 0.001) mean GH levels suppressed by 49 and 80%, respectively. The results support an oral octreotide alternative to parenteral octreotide treatment for patients with acromegaly.

Hsp90 inhibition prevents monocrotaline‐induced pulmonary hypertension in rats, as revealed by high resolution echocardiography

Four weeks after a single sc injection of monocrotaline (MCT; 60mg/kg), rats develop pathologic changes resembling those of human pulmonary arterial hypertension (PAH). Using high resolution echocardiography, we investigated non‐invasively, the ability of the hsp90 inhibitor 17‐AAG (10 mg, 3x weekly) to prevent the effects of MCT. PA flow velocity time integral (VTI), PA acceleration time (PAT), cardiac index (CI) and cardiac output (CO) values were reduced in rats treated with MCT alone, indicative of PAH. 4‐ or 2‐week (starting at week 3 after MCT) treatment with 17‐AAG prevented the decrease in VTI and PAT, but not in CO and CI. Right ventricle (RV) free wall (RVFW) thickness also increased with MCT, but not in the 4‐week 17‐AAG treatment group. Invasive assessment of right ventricular pressure (RVP) at the end of the 4‐week treatments confirmed increased RVP in the MCT, but not in the 17‐AAG treated animals. We conclude that treatment with hsp90 inhibitor for all or part of the 4‐week exposure to MCT effectively prevents the initiation/progression of PAH and that high resolution echocardiography is a useful tool for the non‐invasive, real‐time, repeated evaluation of cardiopulmonary changes in this model of PAH. (Supported by the AHA).

Comparison of Seed Corn Single Crosses SC 704 and SC 770 Response to Different Plant Densities and Nitrogen Levels

An experiment was conducted at the Research Station of Gharakheil of Mazandaran Province, Iran, to evaluate the response of seed corn single crosses SC 704 and SC 770 to different plant densities and nitrogen levels during summer 2010. The soil at the site of the experiment had a loamy texture consisting of 48% sand, 35% silt, and 17% clay, with pH of 7.81, and electrical conductivity of 0.55 mmohs/cm. The experiment was performed in the form of factorial-split using the randomized complete block design. The main factor investigated was the two corn cultivars of Single Cross 704 and Single Cross 770. The sub-factors included three levels of applied nitrogen: the control treatment B1 receiving no nitrogen, treatment B2 receiving 250 Kg nitrogen per hectare and treatment B3 receiving 350 Kg of nitrogen per hectare, along with three plant density levels were C1, C2, and C3, with 75000, 85000, and 95000 plants per hectare, respectively. Results showed that the effect of the cultivar on the trait of grain yield was significant at the probability level of 5 percent. Comparison of the means showed that the cultivar Single Cross 704 with a yield of 12138 Kg.h was superior to the cultivar Single Cross 770 (which had a yield of 10877 Kg.h). Comparison of the means of nitrogen levels indicated that the highest yield (12310 kg.h) was achieved with the application of 350 kg.h of nitrogen and the lowest (10500 kg.h) was obtained in the control treatment. Comparison of the means of the cultivars with respect to the number of seed rows suggested that the cultivar Single Cross 704 with 15.28 seed rows was superior to the cultivar Single Cross 770 with 14.96 seed rows. The rate of nitrogen application had no significant effects on the number of seed rows. The maximum and the minimum numbers of seed rows of 15.74 and 14.48 were observed at densities of 75000 and 95000 plants per hectare, respectively. Results of individual correlation coefficients of the traits studied suggest that seed yield has the strongest positive and significant correlation with the trait of the 1000 – seed weight (0.539*), with the corn ear length (0.52*), and with the number of seed rows per corn ear (0.453*). It was also found that the 1000-seed weight had the strongest positive and significant correlation with the diameter of corn ears (0.700*), the length of corn ears (0.386*), and the number of seed rows per corn ear (0.367*). Furthermore, the number of seeds per corn ear had the strongest positive and significant correlation with the features of the number of seeds per seed row (0.882*), the length of the corn ears (0.737*), and the number of seed rows per ear corn (0.457*).

Unexpected Formation of a Sc3C2@C80 Bisfulleroid Derivative

The reaction of tetrazine 1 with Sc(3)C(2)@C(80) exclusively affords the open-cage derivative 2 instead of the expected C(2)-inserted derivative 3 bearing a four-membered ring, as previously obtained for C(60). The structure of 2 has been firmly established by NMR spectroscopy and theoretical calculations. EPR spectroscopy shows that a single Sc atom of the Sc(3)C(2) cluster gets located within the bulge created by the bridging addend, which is a first step toward release of the internal metal atoms.

227 SUPEROVULATION AND EMBRYO COLLECTION IN WOOD BISON DURING THE ANOVULATORY SEASON

Brucellosis and tuberculosis are endemic in Wood Buffalo National Park, the largest reserve of wood bison (Bison bison athabascae) in Canada. Our goal is to produce and preserve disease-free embryos for the purpose of wood bison repopulation. This study was designed to determine if embryo collection is feasible in wood bison during the anovulatory season (May–July) and to test if progesterone priming is required for superovulation. A 2-by-2 design was used to determine the effectiveness of LH (Lutropin) or hCG (Chorulon) for induction of ovulation with or without intravaginal progesterone releasing device (PRID) in 32 wood bison cows. Follicular wave emergence was synchronized among bison by transvaginal ultrasound-guided follicle ablation. Synchronized bison were assigned to 4 groups: PRID+LH (n = 12), PRID+hCG (n = 4), no-PRID+LH (n = 12) and no-PRID+hCG (n = 4). A PRID was inserted on the day of follicular ablation in the respective groups. A single SC dose of 400 mg FSH (Folltropin) in a slow-release formulation was given the day after follicular ablation (i.e. on the expected day of a new follicular wave emergence, Day 0). The PRID was removed on Day 4 and either 25 mg LH or 2000 IU hCG was given IM on Day 5. Artificial insemination was done at 24, 36 and 48 h after LH or hCG treatment. Embryos were collected nonsurgically on Day 13 using commercial bovine equipment. Transrectal ultrasonography was done on Days 0, 5, 6, 7, 8 and 13 to record follicular and ovulatory responses. Count data (mean ± SEM) were analysed by two-way ANOVA and proportions by chi square. The number of ovulatory-sized follicles (≥10 mm) on Day 5 did not differ among groups (P = 0.33; Table 1). Ovulation rate (number of ovulations/number of follicles ≥10 mm) was greater in bison treated with hCG (P &lt; 0.05; Table 1). The number of corpora lutea (CL) on Day 13 was greater in bison treated with hCG without a PRID (P &lt; 0.05; Table 1). No differences in number of ova/embryos and transferable embryos were found among groups (P = 0.36 and P = 0.52, respectively; Table 1). In conclusion, progesterone priming (PRID) had no effect on ovarian superstimulation in wood bison in the anovulatory season. The ovulatory response was satisfactory only in bison treated with hCG. Embryo collection is feasible in wood bison, but the reasons for a low embryo collection rate in all groups remain unclear. Table 1.Response to superovulation and embryo collection in wood bison Funded by Advancing Canadian Agriculture and Agri-Food and Agri-Food Innovation.

Effects of a Single Dose of Exenatide on Appetite, Gut Hormones, and Glucose Homeostasis in Adults with Prader-Willi Syndrome

Prader-Willi syndrome (PWS) is associated with hyperphagia and obesity, without effective pharmacological treatment. Exenatide, recently developed for treatment of type 2 diabetes, induces appetite suppression and weight loss with common side effects. The objective of the study was to investigate the initial safety and effectiveness of exenatide in adult PWS subjects compared with obese controls (OBESE). Eight PWS and 11 OBESE patients underwent standardized meal studies after a single sc injection of 10 μg exenatide or placebo in a single-blinded, crossover design. Glucose, insulin, C-peptide, glucagon, peptide YY (PYY; total)/PYY (3-36), glucagon-like peptide-1, and ghrelin (total) were measured fasting and postprandially. Appetite and satiety were assessed by visual analog scales. Energy expenditure (EE) was measured by indirect calorimetry. Side effects were screened during and for 24 h after the meal. PWS and OBESE patients were matched for gender, age, body mass index, and central/total body fat. In both groups, exenatide increased satiety and lowered glucose and insulin levels but increased insulin secretion rate. Side effects were absent in PWS but common in OBESE patients. During the meal, PYY (total) and ghrelin were elevated in PWS patients. Exenatide decreased PYY (total) and glucagon-like peptide-1, whereas ghrelin remained unchanged. Energy expenditure was unchanged by exenatide. Our pilot study demonstrates that exenatide is well tolerated in PWS patients. It increases satiety independently of measured appetite hormones, exerting glucose lowering, and insulinotropic effects similarly in PWS and OBESE patients. Larger prospective studies should investigate whether chronic exenatide administration will reduce hyperphagia and overweight in PWS patients without side effects.

Comparison of the efficacy and adverse effects of sustained-release buprenorphine hydrochloride following subcutaneous administration and buprenorphine hydrochloride following oral transmucosal administration in cats undergoing ovariohysterectomy

To compare the efficacy and adverse effects of sustained-release (SR) buprenorphine following SC administration and buprenorphine following oral transmucosal (OTM) administration in cats undergoing ovariohysterectomy. Animals-21 young healthy female cats. As part of anesthetic premedication (0 hours), 10 cats received buprenorphine (0.02 mg/kg) via OTM administration with additional doses at 12, 24, 36, 48, and 60 hours and 11 cats received an equivalent total dose as a single SC injection of SR buprenorphine (0.12 mg/kg). The SR product contained buprenorphine hydrochloride in a proprietary SR matrix. All other anesthetic drugs and a single postoperative dose of meloxicam were administered similarly to all cats. Behavioral and physiologic variables were recorded, and signs of pain were assessed by use of 2 pain assessment scales and von Frey filament testing in each cat prior to premedication administration (baseline), during recovery from anesthesia (RFA), and at 12, 24, 36, 48, 60, and 72 hours. Heart rate increased and temperature (determined via microchip transponder thermometry) decreased from baseline values during RFA in both groups. Compared with baseline values, pain scores were increased during RFA and at the 12- and 24-hour time points in both groups; von Frey scores were higher during RFA. Behavioral and physiologic variables did not differ significantly between groups at any time point. In cats undergoing ovariohysterectomy, SC administration of a preoperative dose of SR buprenorphine appeared to have comparable efficacy and adverse effect profile as that of twice-daily OTM administration of buprenorphine before and after surgery.

Hydrogen storage on scandium-coated toroidal carbon nanostructure C 120 modeled with density functional theory

Ab initio density functional calculations were performed on a toroidal carbon C120 nanostructure doped from one to ten Sc atoms bonded to its outer surface. These calculations are based on DFT with the generalized gradient approximation PW91 (Perdew and Wang) as implemented in the Materials Studio v.4.3 code. The Dmol3 module was used to calculate, among others, total energies, charge density, HOMO-LUMO and Mulliken population analysis. On the basis of these results, it is possible to propose that a single Sc atom is able to adsorb up to 6 H2 molecules. Therefore the study was extended for a system with 10 Sc atoms, which can adsorb up to 60 H2 molecules. This leads to 6.01 wt %, which fulfils the current requirement (6 wt %, at 2010, specified by US Department of Energy (DOE)). Accordingly, the scandium-coated toroidal carbon C120 nanostructure is a good candidate for H2 storage with moderate adsorption energy.

319 SUPEROVULATORY RESPONSE IN JAPANESE BLACK CATTLE BY A SINGLE SUBCUTANEOUS ADMINISTRATION OF PURE FOLLICLE-STIMULATING HORMONE DISSOLVED IN SALINE

The objective of this study was to evaluate the superovulatory response to a single SC administration of pFSH (Antrin R-10) dissolved in saline in Japanese Black cattle. Forty Japanese Black cows were divided into 5 groups. All cows received a progesterone releasing intravaginal device (PRID) at a random stage of the oestrous cycle except the day of oestrus (Day 0). Superovulatory treatments were initiated on Day 4 with single SC administration of 20 AU pFSH dissolved in 10 mL of saline (Group A) or 50 mL of saline (Group B), or 30 AU pFSH dissolved in 10 mL of saline (Group C) or 50 mL of saline (Group D). Conventional superovulatory treatment (Group E) was also initiated on Day 4 with 20 AU pFSH in decreasing doses in IM administration twice a day for 3 days (5/5, 3/3, and 2/2 in the am/pm). Each administration was carried out in the neck region. For oestrus induction, all animals were treated with prostaglandin F2α (750 μg of cloprostenol) on Day 4, and then the PRID was removed on Day 6 in the morning. The GnRH (10 μg of buserelin) was administrated at 31 h after PRID removal to induce ovulations. The cows were artificially inseminated at 24 h after GnRH treatment, and embryos were recovered 7 days after the insemination. Blood samples were collected at 0, 2, 4, 6, 8, 10, 24, 31, 48, 55, 72, and 79 h after the first pFSH administration, and serum pFSH concentration was measured using time-resolved fluorescence immunoassay. Percentage data were arcsine transformed before analysis, and one-way ANOVA was used for statistical analysis (P &lt; 0.05). There were no significant (P &gt; 0.05) differences among groups in the number of corpus luteum, total number of ova/embryos, number of transferable embryos, and percentage of transferable embryos. The concentrations of pFSH in serum increased in all groups 2 h after each administration and reached a peak at 8 h in Groups A and C, at 10 h in Groups B, D, and E, and then in all groups it decreased gradually. The concentrations of pFSH in serum decreased to the base level (the level at 0 h) in Group A at 55 h and in Group C at 72 h after the first pFSH administration. At 79 h, serum pFSH was still detected in Groups B, D, and E. These results suggest that the single SC administration of 20 AU pFSH that is the same dose as the conventional treatment is able to induce a superovulatory response comparable to that of conventional treatment in Japanese Black cattle. Table 1.Superovulatory responses (mean ± SE) in different treatment groups

Increased aromatase expression in the hippocampus of spontaneously hypertensive rats: effects of estradiol administration

There is high incidence of hippocampal abnormalities in spontaneously hypertensive rats (SHR), including decreased neurogenesis in the dentate gyrus, astrogliosis, low expression of brain derived neurotrophic factor and decreased neuronal density in the hilar region, respect of normotensive Wistar Kyoto rats (WKY). Estradiol treatment given for 2 weeks normalized the faulty hippocampal parameters of SHR, without having effects on WKY rats. The present work studied the potential role of local estrogen biosynthesis in the hippocampus of SHR and WKY, by measuring the expression of aromatase, the key enzyme responsible for estrogen biosynthesis and involved in neuroprotection. We used 4 month old male SHR and WKY, half of which received a single sc pellet of 12 mg estradiol benzoate and the remaining half a cholesterol implant. Hippocampi were dissected and processed for aromatase mRNA expression using real time PCR. A second batch of animals was processed for aromatase and glial fibrillary acidic protein (GFAP) immunocytochemistry. Basal level of aromatase mRNA was higher in SHR respect of WKY. Following estradiol treatment, aromatase mRNA was further increased in the SHR group only. In the hilus of the dentate gyrus of cholesterol-implanted SHR, we found aromatase immunoreactive cell processes and fibers more strongly stained respect of WKY rats. Estradiol treatment of SHR further increased the length of immunoreactive processes and fibers in the hilar region and also increased aromatase immunoreactivity in the CA1 but not the CA3 pyramidal cell region. WKY rats were spared from the estradiol effect. Double-labelling experiments showed that aromatase+ processes and fibers of the hilus of SHR-treated rats did no colocalize with GFAP+ astrocyte cell bodies or processes. In conclusion, basal and estradiol-stimulated aromatase expression was enhanced in hypertensive rat hippocampus. A combination of exogenous estrogens and those locally synthesized may better alleviate hypertensive encephalopathy.

Growth and scintillation properties of Sc, Pr, Ce co-doped LuAG epitaxial garnet layers

Single crystalline Sc, Pr, Ce co-doped Lu3Al5O12 garnet layers were grown by liquid phase epitaxy. The Sc doping increases the scintillation response of Pr3+ activator ions because of overlap of the Sc-related emission around 275 nm with the 4f→5d absorption band of Pr3+ centers. The radioluminescence (RL) spectra give evidence for presumed energy transfer form Sc3+ to Pr3+ activator ions. Significant increase of the RL emission output was observed when co-doping with both Pr and Sc ions compared to Sc-free samples. The maximum integral RL was noticed for samples with relatively high Sc concentration, 1.2 - 3 at %. On the contrary, an increase of the photoelectron yield, Nphe, measured under alpha particle excitation, was observed only for rather low Sc content, ∼0.6 at %, at higher Sc concentrations Nphe decreased. Nevertheless, LuAG epitaxial layers with optimal Sc and Pr contents show comparable or even a bit higher Nphe yield and energy resolution than Cz-grown Pr:LuAG single crystals. Furthermore, the slow components in photoelectron response were significantly suppressed in epitaxial layers compared to single crystals.