1X Group Research Articles

Effects of Chlorich®EnergyBoost on Enhancing Physical Performance and Anti-Fatigue Properties in Mice.

Chlorich®EnergyBoost, a water extract obtained from Chlorella sorokiniana, has been proposed to enhance physical performance and provide anti-fatigue effects. This study assessed the impact of Chlorich®EnergyBoost supplementation on physical performance and its anti-fatigue properties. Twenty-four mice were allocated into four groups: (1) the control group receiving only water,;(2) the 1X group (49.2 mg/kg/day); (3) the 2X group (98.4 g/kg/day); and (4) the 5X group (246 g/kg/day). All groups were orally administered the supplements for four consecutive weeks. The evaluation included grip strength, swimming endurance, an exhaustion test, and serum biochemistry analysis. Additionally, the study examined the bioactive peptides through matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) and conducted bacterial reverse mutation and acute oral toxicity tests for safety assessment. The findings indicated that Chlorich®EnergyBoost supplementation led to a significant reduction in serum lactate levels by 14.08% to 22.54% and blood urea nitrogen levels by 12.23% to 16.76%, an increase in the lactate clearance rate by 0.28 to 0.35, an enhancement of muscle glycogen storage by 1.10 to 1.44-fold, and hepatic glycogen storage by 1.41 to 1.47-fold. These results demonstrated dose-dependent effects. MALDI-TOF analysis revealed the expression of dihydrolipoamide dehydrogenase and superoxide dismutase. Both the bacterial reverse mutation and acute oral toxicity tests showed no adverse effects.

Histopathological changes and tissue residue concentrations of monosex Nile tilapia (Oreochromis niloticus, L) fries exposed to oxytetracycline.

The current study evaluated the biosafety of oxytetracycline (OTC) exposure for 30 days in monosex Oreochromis niloticus fries. The fries were exposed to OTC for 3 h/day for 30 days at 350 (0.5X), 700 (1X), 2100 (3X), 3500 (5X), and 7000 (10X) mg/L and compared with control (0X). The OTC exposure at 5X and 10X concentrations caused 100% mortality within 4 days and 5 min, respectively. The mortalities recorded in 0.5X, 1X, and 3X groups were 3.33 ± 1.15%, 14.67 ± 1.15%, and 47.33 ± 11.37% on day 30, respectively. The feed intake was decreased up to 23.33% in the 3X group during the exposure period. The OTC residue levels on 30-day exposure were 216.53 ± 14.71, 450.56 ± 44.31, and 1141.26 ± 63.64 μg/kg, which reduced to 40.40 ± 3.25, 76.68 ± 2.77, and 95.61 ± 5.13 μg/kg after 15 days of termination of exposure in the 0.5X, 1X, and 3X groups, respectively. The histopathological changes observed in the 1X group were epithelial detachment, desquamation of secondary lamellar epithelium, lamellar fusion, and inflamed cartilaginous core in the gills, alteration in the integrity of gut mucosa, degeneration of muscularis mucosae and necrosis in the intestine, the disintegration of the nephritic tubule, necrosis, and glomerulopathy in the kidney, and dilated vascular duct, necrotized hepatic tissue, diffused hepatic parenchyma, vacuolation, and fatty changes in the liver. The OTC exposure induced marked tissue changes histologically in a dose- and time-dependent manner, which undoubtedly reduced the growth of tilapia.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10499-022-00892-w.

Influence of dietary emamectin benzoate on the biological responses of monosex (all-male) Oreochromis niloticus (L.) fries

The application of antiparasitic drugs plays a crucial role in the removal of infectious parasites in aquaculture. Emamectin benzoate (EB) is predominantly used as a feed premix against ectoparasites on temperate fish. This study evaluated the influence of 14 days of EB-dosing at 0–10 times the recommended dose (1X: 50 μg/kg biomass/day) on the biological responses and accrual/depletion of EB-residues in a tropical fish monosex Oreochromis niloticus fries. A significant dose-dependent reduction in feed intake by 3.50% in 1X and 43.00% in 10X groups, and an increase in mortalities from 2.92% (1X) to 11.25% (10X) during the EB-dosing period was noted. A significant increase in glucose and alkaline phosphatase and reduction in calcium and chloride ions, superoxide dismutase (SOD) and acetylcholinesterase levels in the muscle and/or brain tissue was observed. On day 21 post-EB-dosing, the levels of muscle glucose and SOD reached normalcy in the 1X group, while the levels of other biomarkers failed to recuperate. The EB-residue levels peaked on day 14 EB-dosing (2.77 ng/g) in the 1X group and decreased later with detectable levels (0.03 ng/g) even on day 21 post-EB-dosing. The EB-residue levels were within the permissible limits of the Canadian Food Inspection Agency and the European Commission. The EB-dosing negatively influenced the health of O. niloticus by altering the physiological state in a dose- and time-dependent way. The results suggested that the use of EB might be plausibly risky in tropical aquaculture.

Dietary influences of oxytetracycline on the growth and serum biomarkers of Oreochromis niloticus (L.)

Tilapias are cultured globally and are rising in acceptance as the most important freshwater aquaculture species. Monitoring of serum biomarkers is a promising tool in aquaculture to screen the health status as they are virtuous indicators of extreme stress and organ dysfunction in fish. The present study examined the serum biomarkers of oxytetracycline (OTC)-dosed Nile tilapia Oreochromis niloticus at 0, 80 and 800 mg/kg biomass/day, i.e., 0X, 1X, and 10X the approved dose (X = 80 mg OTC/kg biomass/day) for 10 consecutive days. The fish biomass and levels of serum glucose, aspartate aminotransferase, alanine aminotransferase, creatinine and C-reactive protein (CRP) were determined at scheduled intervals. A significant dose-dependent reduction in fish biomass during the OTC-dosing (5.84%) and post-OTC dosing (8.16%) periods was observed. All the serum biomarkers of Nile tilapia increased significantly on day 10 OTC-dosing. Though their levels reduced significantly, normalcy was not achieved even after 42 days of cessation of OTC-dosing, except CRP. The CRP reached the normal level on day 25 post-OTC dosing in the 1X group. The results, thus, demonstrated that the oral OTC-dosing influences the physiological state of apparently healthy Nile tilapia in a dose-dependent manner. These changes were, however, reversible upon discontinuation of OTC-dosing. The set of data observed on growth reduction and elevated serum biomarker levels even after 42 days of cessation of OTC-dosing, thus, raises questions on the utility of oral OTC-dosing.

Effects of Antrodia camphorata and Panax ginseng supplementation on anti-fatigue properties in mice.

Antrodia camphorata and Panax ginseng are well-known medicinal plants in Taiwan folk and traditional Chinese medicine, which have been reported for multifunctional bioactivities. However, there is limited evidence that a fixed combination formula of these two plant extracts is effective for the exercise improvement or anti-fatigue. We aimed to evaluate the potential beneficial effects of the mix formulation of these two herbal medicines (AG formulation) on fatigue and ergogenic functions following physiological challenge. Male Institute of Cancer Research (ICR) mice from four groups (n=10 per group) were orally administered AG formulation for 4 weeks at 0.984, 2.952 and 5.904 g/kg/day, which were designated the Vehicle, AG-1X, AG-3X and AG-6X groups, respectively. The anti-fatigue activity and exercise performance were evaluated using exhaustive swimming time, forelimb grip strength, and levels of serum lactate, ammonia, glucose, blood urea nitrogen (BUN) and creatine kinase (CK) after a swimming exercise. The exhaustive swimming time of the 1X, 3X or 6X AG group was significantly longer than that of the Vehicle group, and the forelimb grip strength of the 1X, 3X or 6X AG group was also significantly higher than that of the Vehicle group. AG supplementation also produced decreases in serum lactate, ammonia, BUN and CK activity after the swimming test, as well as increases in glucose. Therefore, the AG complex could be a potential formulation with an anti-fatigue pharmacological effect.

Repeat dose NRPT (nicotinamide riboside and pterostilbene) increases NAD+ levels in humans safely and sustainably: a randomized, double-blind, placebo-controlled study

NRPT is a combination of nicotinamide riboside (NR), a nicotinamide adenine dinucleotide (NAD+) precursor vitamin found in milk, and pterostilbene (PT), a polyphenol found in blueberries. Here, we report this first-in-humans clinical trial designed to assess the safety and efficacy of a repeat dose of NRPT (commercially known as Basis). NRPT was evaluated in a randomized, double-blind, and placebo-controlled study in a population of 120 healthy adults between the ages of 60 and 80 years. The study consisted of three treatment arms: placebo, recommended dose of NRPT (NRPT 1X), and double dose of NRPT (NRPT 2X). All subjects took their blinded supplement daily for eight weeks. Analysis of NAD+ in whole blood demonstrated that NRPT significantly increases the concentration of NAD+ in a dose-dependent manner. NAD+ levels increased by approximately 40% in the NRPT 1X group and approximately 90% in the NRPT 2X group after 4 weeks as compared to placebo and baseline. Furthermore, this significant increase in NAD+ levels was sustained throughout the entire 8-week trial. NAD+ levels did not increase for the placebo group during the trial. No serious adverse events were reported in this study. This study shows that a repeat dose of NRPT is a safe and effective way to increase NAD+ levels sustainably.

11 COMPARISON OF CONTROLLED INTERNAL DRUG-RELEASE INSERT–BASED AND PROGESTERONE-FREE METHODS FOR OVULATION SYNCHRONIZATION AND TIMED ARTIFICIAL INSEMINATION OF GOATS

A CIDR synchronization program is an important tool for facilitating ovulation synchronization and timed AI (OvSynch-TAI). The objective of this study was to test the efficacy of reusing CIDR for OvSynch-TAI compared with that for a progesterone-free OvSynch-TAI protocol (NCS) or a breed by oestrus detection (ED) control. A total of 87 does were randomised into 1 of 5 treatments: (1) ED (control, n = 18), (2) NCS (n = 18), (3) CIDR6-New (n = 17), (4) CIDR6–1X (n = 17), and (5) CIDR6–2X (n = 17). Does in the ED group received two 15-mg doses of PGF2α at a 10-d interval and were bred 12 h after the onset of oestrus following the second PGF2α injection. The NCS group received 15 mg of PGF2α on Day 0, 50 μg of gonadotropin-releasing hormone on Day 8, 15 mg of PGF2α on Day 15, and 50 μg of gonadotropin-releasing hormone on Day 18, concurrently with TAI. The CIDR groups (new, 1X-used, or 2X-used) received a P4 device for a 6-d period, and 15 mg of PGF2α was administered at CIDR removal. Does were bred 48 h after CIDR removal and were given 50 μg of gonadotropin-releasing hormone at TAI. The CIDR in the CIDR6–1X group were previously in place for 10 days before use, whereas CIDR in the CIDR6–2X group were previously in place for 16 days before use. Reused CIDR were rinsed in a diluted Nolvasan solution, followed by a clean water rinse, allowed to air dry, and stored in a refrigerator until time of use. The experiment was conducted in 2 replicates. Within each replicate, all treatments were scheduled so that does were bred during the same 2-day period, and all does were inseminated with a single dose of frozen semen using a nonsurgical (transcervical) technique. Blood samples were taken daily in all treatment groups to monitor concentrations of serum progesterone until the time of breeding. Pregnancy was determined by ultrasonography at 54 and 85 days of gestation. Data were analysed using GLM procedures of SAS (SAS Institute Inc., Cary, NC, USA). Data for pregnancy rates were analysed with a model that included effects of treatment, replicate, and their interactions. Data for serum progesterone concentrations were analysed with a model that included the effects of treatment, replicate, day, and their interactions. Means were separated by Duncan’s test. Mean progesterone differed (P < 0.001) with treatment (5.3 ± 0.8bc, 3.5 ± 0.8c, 7.0 ± 0.8ab, 7.9 ± 0.8a, 6.2 ± 0.8ab ng mL–1 for ED, NCS, CIDR6-New, CIDR6–1X, and CIDR6–2X, respectively; least squares mean ± standard error of the mean). Pregnancy rates for the ED, NCS, CIDR6-New, CIDR6–1X, and CIDR6–2X treatment groups were 39 ± 11%bc, 22 ± 11%c, 64 ± 12%ab, 77 ± 12%a, and 57 ± 12%ab, respectively. In conclusion, reused CIDR were as effective as new CIDR for attaining satisfactory pregnancy rates. Timed AI using a once-used CIDR was more effective for establishing pregnancy than ED and NCS treatments. The lower pregnancy rates in the ED and NCS groups were associated with lower mean progesterone levels during the Ovsynch treatment period before breeding. This research was supported by the North Carolina Agricultural Experiment Station.

Evaluation of implant strategies in Angus-sired steers with high or low genetic potential for marbling and gain1

Sixty-nine Angus-sired steer calves (332.3 kg initial BW) were used to determine the effects of single or double implant strategies on steers of high or low genetic potential (GP) determined by the GeneMax (Zoetis, Florham Park, NJ) genetic profiling test. Steers were assigned to treatments in a 2 × 2 factorial design with factors of 1) composite GP score (high, mean GP score of 86.5 [HI]; low, mean GP score of 25.3[LO]) and 2) implant strategy (single, steers implanted on d 70 [1X], or double, steers implanted d 0 and 70 [2X]). All steers were given the same implant (Revalor-S; Merck Animal Health, Summit, NJ), with the 2X group implanted on d 0 and 70 and the 1X group implanted only on d 70. A diet containing 1.38 Mcal NEg/kg DM was fed ad libitum, once daily. Ultrasound was used to measure body composition characteristics on d 0 and 70. Steers were harvested after 140 d on feed. At both the d-0 and d-70 ultrasound, HI steers had greater ( < 0.001) percent intramuscular fat (IMF) than LO steers, but no differences ( ≥ 0.24) were observed in LM area (LMA), rib fat thickness (RF), or rump fat thickness (RMFT). Steers in the 2X group had larger ( = 0.02) LMA and less ( = 0.03) IMF on d 70 than 1X steers and no differences ( ≥ 0.50) in RF or RMFT were observed. From d 0 to 70, HI steers had ADG, DMI, and G:F ( ≥ 0.60) similar to LO steers; however, 2X steers had greater ( < 0.001) ADG and were more ( < 0.001) feed efficient compared with 1X steers during the same interval. Over the entire 140-d feeding period, there were no differences ( ≥ 0.6) in BW, ADG, DMI, or G:F between GP groups; however, 2X steers had greater ( = 0.03) ADG compared with 1X steers and still had similar ( ≥ 0.12) DMI and G:F. Upon slaughter, marbling score tended to be impacted by a GP × implant interaction (499.9 ± 18.5, 545.6 ± 18.5, 487.1 ± 18.5, and 469.8 ± 18.5 for HI and 2X, HI and 1X, LO and 2X, and LO and 1X, respectively; = 0.06). No differences ( ≥ 0.7) were observed between GP groups for HCW, LMA, RF, KPH, or yield grade (YG). Steers in the 1X group had less ( = 0.003) RF than 2X steers but similar ( ≥ 0.14) HCW, marbling, LMA, KPH, and YG. A greater proportion ( = 0.03) of HI steers had choice carcasses (100 ± 0.0%) compared with LO steers (87.8 ± 3.9%). Results of this study indicate that the GP test used in the current study predicted differences in IMF, carcass marbling, and percent carcasses graded as choice.

Tiger, Bengal and Domestic Cat Embryos Produced by Homospecific and Interspecific Zona-Free Nuclear Transfer.

The aim of this study was to evaluate three different cloning strategies in the domestic cat (Felis silvestris) and to use the most efficient to generate wild felid embryos by interspecific cloning (iSCNT) using Bengal (a hybrid formed by the cross of Felis silvestris and Prionailurus bengalensis) and tiger (Panthera tigris) donor cells. In experiment 1, zona-free (ZP-free) cloning resulted in higher fusion and expanded blastocyst rates with respect to zona included cloning techniques that involved fusion or injection of the donor cell. In experiment 2, ZP-free iSCNT and embryo aggregation (2X) were assessed. Division velocity and blastocyst rates were increased by embryo aggregation in the three species. Despite fewer tiger embryos than Bengal and cat embryos reached the blastocyst stage, Tiger 2X group increased the percentage of blastocysts with respect to Tiger 1X group (3.2% vs 12.1%, respectively). Moreover, blastocyst cell number was almost duplicated in aggregated embryos with respect to non-aggregated ones within Bengal and tiger groups (278.3 ± 61.9 vs 516.8 ± 103.6 for Bengal 1X and Bengal 2X groups, respectively; 41 vs 220 ± 60 for Tiger 1X and Tiger 2X groups, respectively). OCT4 analysis also revealed that tiger blastocysts had higher proportion of OCT4-positive cells with respect to Bengal blastocysts and cat intracytoplasmic sperm injection blastocysts. In conclusion, ZP-free cloning has improved the quality of cat embryos with respect to the other cloning techniques evaluated and was successfully applied in iSCNT complemented with embryo aggregation.

Abstract 1631: Bioluminescence imaging enhances analysis of drug response in a patient-derived xenograft model of pediatric acute lymphoblastic leukemia

Abstract Despite the success of current therapy, 15-20% of pediatric acute lymphoblastic leukemia (ALL) patients fail treatment. Since the number of new agents for testing outweighs the availability of patients for clinical trials, robust preclinical models are required to prioritize effective drugs. Patient-derived xenografts (PDXs) of ALL in immune-deficient mice are a clinically relevant testing model as they recapitulate original disease characteristics and allow monitoring of engraftment and drug response by flow cytometric enumeration of the% human vs mouse CD45+ cells in the mouse peripheral blood (PB). However, factors such as leukemic harbor and relapse sites are overlooked, and this model may be biased towards drugs that clear circulating lymphocytes. This study aimed to overcome these shortfalls by introducing a bioluminescence imaging (BLI) capability into a pediatric ALL PDX to enhance monitoring of leukemia burden. Firefly luciferase was incorporated via lentiviral transduction, and BLI and PB measures used to track engraftment and response to an induction-type drug regimen; vincristine (0.15 mg/kg weekly x 4), dexamethasone (5 mg/kg daily x 5 × 4) and L-asparaginase (1,000 KU/kg daily x 5 × 4) (VXL). Additional groups received 0.5, 0.25 and 0.125x this dose for 2 weeks. Engraftment by BLI and PB correlated strongly (R2 = 0.8, p = &amp;lt;0.0001) yet BLI detected engraftment 6 weeks before PB. VXL response was scored using PB measures and stringent response criteria used by the Pediatric Preclinical Testing Program, modeled after the clinical setting. The 0.125, 0.25, 0.5 and 1x groups scored Progressive Disease 2, Stable Disease, Complete Response (CR) and Maintained CR, respectively. BLI was able to quantify regression with greater range and sensitivity than PB, highlighting a large difference in regression between the 1x and 0.5x groups. A median 3.2-fold decrease in whole-body bioluminescence (photons/sec/steradian/cm2) was seen in the 0.5x group at treatment end relative to Day 0, compared to a &amp;gt;2,000-fold decrease in the 1x group. Autopsy of mice at Day 28 revealed a median of 0.8% blasts in the bone marrow (BM) in the 1x group compared to 35% in the 0.5x group. Since the clinical definition of a CR is &amp;lt;5% blasts in the BM at Day 28, the 0.5x group did not achieve a CR by this classification, despite scoring a CR by PB measurements. BLI supported this finding, revealing &amp;gt;2000-fold decrease in BM bioluminescence in the 1x group relative to Day 0, and a 1.3-fold increase in the 0.5x group. These results show that BLI enhances the sensitivity of drug efficacy testing in this PDX model, and allows quantification of efficacy in discreet leukemic harbor sites such as the BM. This work suggests BLI can increase the stringency of preclinical drug testing to accurately prioritize agents for clinical trials. Citation Format: Luke Jones, Jennifer Richmond, Kathryn Evans, Richard B. Lock. Bioluminescence imaging enhances analysis of drug response in a patient-derived xenograft model of pediatric acute lymphoblastic leukemia. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1631. doi:10.1158/1538-7445.AM2015-1631

72 EFFECTS OF A PRECONCEPTIONAL AND GESTATIONAL MULTI-VITAMIN-MINERAL-OMEGA3 SUPPLEMENTATION ON FETOPLACENTAL DEVELOPMENT IN A RABBIT MODEL

The developmental origins of health and disease concept stipulates that nutritional imbalance in the preconceptional and gestational periods induces risks for the development and long-term health of the individual. In industrialized countries, most women take nutritional supplements before and during pregnancy, regardless of their diet. Potentially harmful embryonic and fetal effects of an excess multi-micronutrient supplementation, however, are not known. The objective of this study was to assess, using a rabbit model, the effects of a multi-vitamin-mineral-omega 3 supplementation (Gestarelle® G) administered in slight excess during preconceptional and gestational periods. Twenty-seven New Zealand does were used. Supplementation contained vitamins (B1, B2, B5, B6, B8, B9, B12, C, E), minerals (magnesium, iron, copper, iodine), and n-3 polyunsaturated fatty acids (n-3 PUFA; namely DHA and EPA) in the form of oral capsules administered daily from 10 days before mating until the end of pregnancy. Group 1X posology allometrically matched that used in humans (n = 10), representing an excess of 6 to 50% of rabbit needs. Group 3X rabbits received 3 times the recommended posology (n = 9; 18 to 150% of rabbit needs), whereas the control group received capsules containing only excipient (n = 8). Embryo and fetoplacental development was monitored by transabdominal two- and three-dimensional ultrasound and Doppler on Days 5, 7, 14, 21, and 28 with 5 conceptuses per doe examined each time. Does were killed on Day 28 (term = 31 days), 24 h after the last capsule administration. Maternal and fetal blood, as well as placenta and fetal organs, were collected. Data were analysed by ANOVA and Kruskall-Wallis as appropriate. The n-3 PUFA plasma concentrations increased in the 3X does (P &lt; 0.01). Fetal plasma concentrations were increased only in males in the 1X group (P &lt; 0.001) and both in females and males in the 3X group (P &lt; 1e–15). No statistical differences were observed between treatments for any of the ultrasound or Doppler analyses. In total, 208 fetuses were collected with no difference in litter size, sex ratio, nor biometric measurements between groups. No structural anomaly was observed by histological analysis of fetal organs. In conclusion, multi-vitamin-mineral-omega 3 supplementation administered during the preconceptional and gestational periods in a rabbit model in the present study and within the measured parameters had no deleterious effect for dam and conceptuses at 1 and 3 times the recommended posology. Further work is on-going to study the effects on fetal brain development. Further studies are needed to evaluate putative post-natal effects. This study was funded by IPRAD Santé Laboratories, Paris, France.

EFFECTS OF MILKING FREQUENCY ON MILK PRODUCTION AND HISTOLOGICAL STRUCTURE OF UDDER IN ZARAIBI DAIRY GOATS

ABSTRACTThis work was carried out on 32 Egyptian Nubian (Zaraibi) goats to investigate both the effect of the milking frequency on milk yield and milk composition and the effect of stage of lactation on histological structure and histochemistry of the secretory mammary cells and its relationship with milk production in Zaraibi goats. Biopsies were taken surgically from the mammary gland from 3 does milked once daily (1x) and 3 does milked twice daily (2x) at the three stages of lactation, early, mid and late, for histological and histochemical studies. The histological structure showed clear differences due to milking frequency and lactation stage, being more developed at early and mid stages and twice daily milking compared to late stage of lactation and once daily milking. The number of the alveolus secretory cells per alveolus increased from the early to the mid stage of lactation by17.6% and then reduced at the late stage by 25% compared to mid stage, while no difference noticed between twice and once daily milking. Once daily milking (1x) reduced milk yield by 6%, and increased fat percentage to 4.0% compared to 3.67% in twice daily milking group (2x). Milk of once milking (1x) group contained higher percentage of total solids 11.38% than twice milking (10.93%), but milk protein and lactose did not differ between 1x and 2x milking. Lactation curve showed 32.2% increase in yield during mid stage than early stage, while late stage attained 61.3% reduction in milk yield compared to mid stage. Protein and lactose percentages did not change throughout different stages of lactation, while fat and total solids percentages showed the highest values at early stage of lactation (4.0 and 11.5%, respectively), and the lowest at mid and late stage of lactation. The total sectional areas (u/plate) of the alveoli were the smallest during late lactation (495399 u /plate) compared to that during early and mid stages of lactation (705206 and 759901u/plate, respectively). Numerous loci of alcaline phosphatase (AP) were apparent on the outer surface of the alveolar secretory cells at the early and mid stages of lactation—reflecting high activity of this enzyme at these two stages. This was accompanied by a high level of milk secretion reaching1778.2±38.9 and 2351.4±68.4 g/head/day, respectively.In contrast, at the late stage of lactation, the size of alveoli was reduced and few alveoli showed weak AP activity. This coincided with the reduction in milk yield (910g/head/day). It could be concluded that stages of lactation influence the cell number and activity of mammary epithelial cells.

Effects of Zinc‐l‐Carnosine and Vitamin E on Aspirin‐Induced Gastroduodenal Injury in Dogs

Nonsteroidal anti-inflammatory drugs frequently cause gastrointestinal (GI) injury. Zinc-L-carnosine has antioxidant, anti-inflammatory, mucosal protective, and healing properties in rodent models and in some human studies of GI injury. The combination of zinc-L-carnosine and vitamin E attenuates aspirin-induced gastroduodenal mucosal injury. Eighteen healthy random-source Foxhound dogs. In this randomized, double-blinded, placebo-controlled study dogs were treated with placebo (n = 6; 0X group), 30 mg/30 IU (n = 6; 1X group), or 60 mg/60 IU (n = 6; 2X group) zinc-L-carnosine/vitamin E orally every 12 hours for 35 days. Between Day 7 and 35, GI mucosal lesions were induced with aspirin (25 mg/kg p.o. q8h). Mucosal injury lesions (hemorrhage, erosion, and ulcer) were assessed by gastroduodenoscopy on Days 14, 21, and 35 with a 12-point scoring scale. At baseline (Day -1) gastroscopy scores were not significantly different between groups (mean ± SD: 0X, 4.4 ± 0.8; group 1X, 4.4 ± 0.6; group 2X, 4.2 ± 0.3; P= .55). Gastroscopy scores increased significantly in all groups between Day -1 and Days 14, 21, and 35 (P < .0001). On Day 35, gastroscopy scores were 29.2 ± 5.2 (0X), 27.3 ± 3.7 (1X), and 28.6 ± 3.3 (2X). Mean gastroscopy scores were not significantly different among treatment groups on any of the days (P = .61). Administration of the combination of zinc-L-carnosine and vitamin E at 1X or 2X dosing did not attenuate aspirin-induced gastroduodenal mucosal injury.

Effect of Docosahexaenoic Acid on Quality of Cryopreserved Boar Semen in Different Breeds

During the cryopreservation process, the level of polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), in the sperm plasma membrane decreases significantly because of lipid peroxidation, which may contribute to sperm loss quality (i.e. fertility) of frozen-thawed semen. The aim of this study was to investigate the effect of supplementation of DHA (fish oil) in freezing extender II on frozen-thawed semen quality. Semen from 20 boars of proven motility and morphology, were used in this study. Boar semen was split into four groups, in which the lactose-egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various levels of fish oil to reach DHA level of 1X (group I, control, no added fish oil), 6X (group II), 12X (group III) and 18X (group IV). Semen solutions were frozen by using a controlled rate freezer. After cryopreservation, frozen semen was thawed and evaluated for progressive motility, viability by using SYBR-14/Ethidiumhomodimer-1 (EthD-1) staining and acrosome integrity by using FITC-PNA/EthD-1 staining. There was a significantly higher (p < 0.001) percentage of progressive motility, viability and acrosome integrity in DHA (fish oil) supplemented groups than control group. Generally, there seemed to be a dose-dependent effect of DHA, with the highest percentage of progressive motility, viability and acrosome integrity in group-III. In conclusion, supplementation of the LEY extender with DHA by adding fish oil was effective for freezing boar semen as it resulted in higher post-thaw plasma membrane integrity and progressive motility.

Long-term metabolic effects of different doses of niacin-bound chromium on Sprague-Dawley rats

We simultaneously assessed benefits and risks of niacin-bound chromium (NBC) intake at varying doses over a prolonged period of time (>1.2 years) in male and female Sprague-Dawley (SD) rats. We performed the study in two phases. First, we followed 60 male and 60 female SD rats, each gender divided into six groups. Through day 150 (phase 1A), all SD rats received a high sucrose diet (30% w/w) with or without different concentrations of NBC. The male/female groups were: 1] control without NBC n = 10, 2] low NBC (2.8 ppm, n = 10), 3] medium NBC (8.7 ppm, n = 20), 4] high NBC (28.0 ppm, n = 20). Based on dosing, we refer to the three treatment groups as 1X, 3X, and 10X. During days 151-312 (phase 1B), NBC was removed from diets of one half of the 3X and 10X groups. These are referred to as 3X satellite and 10X satellite. In phase 2 (days 313-460), males from groups 1X, 3X, 10X, 3X satellite, and 10X satellite received the same 3X dose of NBC (8.7 ppm). The last two groups also ingested different doses of a formulation of natural products in addition to NBC. We examined blood pressure, the renin-angiotensin system (RAS), nitric oxide (NO), and insulin systems and inflammatory parameters. Results in male and female SD rats were comparable. NBC lowered systolic blood pressure (SBP) in a dose-dependent fashion; however, after 200 days, the SBP of the low dose group (1X) began to rise and returned to baseline control. After raising the dose of NBC to 3X, the SBP in the 1X group decreased significantly once more. When half the test rats (3X and 10X) were deprived of NBC, SBP rose gradually to control levels after 2 to 3 months. However, the SBP decreased significantly once more when each satellite group returned to the 3X dose. Special testing suggests that NBC at adequate dosing increases insulin sensitivity, lowers HbA1C, decreases activity of the RAS, at least in part, through ACE inhibition, enhances NO activity, and is without signs of toxicity. The addition of a formula composed of antioxidants and immune modulators to the chromium regimen caused even faster and more profound changes in SBP than with NBC alone. We conclude that NBC at adequate dosing is effective in male and female SD rats on certain metabolic parameters over a prolonged period, effects that disappear over months after NBC is removed. When dosing is returned, the effectiveness of NBC returns. Low doses of NBC may lose their effect over time. No signs of toxicity were observed.

Intraperitoneal administration of adenosine inhibits formation of abdominal adhesions.

Previously we demonstrated that peritoneal lavage with high concentrations of adenosine (1 mM) provides pharmacologic levels of adenosine in the intestines without elevating adenosine levels in the systemic circulation and without causing systemic hemodynamic effects (Alimentary Pharmacology & Therapeutics 2000; 14:1371-80). Because adenosine can be safely administered into the peritoneal cavity, and because it inhibits fibroblast proliferation and collagen production and inflammation and enhances angiogenesis, we tested the hypothesis that adenosine applied into the abdominal cavity safely and effectively reduces formation of abdominal adhesions. To test this hypothesis, in Sprague-Dawley rats, a window of right parietal peritoneum was removed and the cecum was brushed and placed next to the damaged peritoneum. After injury, rats received in the abdominal cavity either 20 ml of saline (n = 12, Saline Group) or 20 ml of 1 mM adenosine (n = 12, Adenosine 1X Group; and n = 12, Adenosine 4X Group). At 24, 48, and 72 hours after surgery, rats received by intraperitoneal injection either 10 ml of saline (Saline Group and Adenosine 1X Group) or 10 ml of 1 mM adenosine (Adenosine 4X Group). After 14 days, the degree of adhesion formation was scored (0 to 4) by a blinded observer. Animals tolerated the adenosine treatments without signs of discomfort or distress. The adhesion scores were 2.6 +/- 0.34, 1.7 +/- 0.40, and 0.74 +/- 0.29 in the Saline, Adenosine 1X, and Adenosine 4X groups, respectively (P = 0.0035, Kruskal-Wallis analysis of variance). Peritoneal administration of 1 mM adenosine safely and effectively reduces adhesion formation.

Effect of frequency of boar exposure and adjusted mating times on measures of reproductive performance in weaned sows.

In weaned sows, reduced reproductive performance can result from failure or delayed return to estrus or improper timing of insemination. This experiment evaluated the effect of increased frequency of boar exposure and adjusted mating times on reproductive performance. Sows of mixed parity were weaned approximately 18.7 d after parturition and allotted by genotype, parity, and lactation length to boar exposure frequency of once daily (1X, n = 66), twice daily every 12 h (2X, n = 61), or three times daily exposure at 8-h intervals (3X, n = 60). Sows were weaned into crates and boar exposure was initiated 3 d after weaning. Once estrus was detected, ultrasound was performed every 8 h to determine time of ovulation. All sows were artificially inseminated twice in the 1X group at 0 and 24 h, in the 2X group at 12 and 24 h, and in the 3X group at 16 and 32 h after onset of estrus. The weaning-to-estrus interval was not influenced by treatment and averaged 4.5 d. The percentage of sows expressing estrus in 8 d was higher (P < 0.05) for 1X (97.3%) compared with 2X (92.8%) but not the 3X group (94.0%). The percentage of sows ovulating after estrus was not influenced by treatment (P > 0.10) and averaged 96.5%. Estrus-to-ovulation interval was not affected by treatment (44.7 h) but was influenced by weaning-to-estrus interval (P < 0.0001). Length of estrus was influenced by treatment (P < 0.001), with estrus in the 1X (46.6 h) shorter than in the 2X (60.0 h) and 3X (67.0 h) treatments, and also by weaning-to-estrus interval (P < 0.001). The percentage of first inseminations occurring within 24 h before ovulation was increased (P < 0.001) in the 2X (62%) and 3X (66%) groups compared with the 1X group (28%) and was also influenced by parity (P < 0.001) and weaning-to-estrus interval (P < 0.05). The percentage of second services within 24 h before ovulation was not increased by any factor and averaged 78%. Farrowing rates were not increased (P > 0.10) for 2X (87.2%) and 3X (83.1%) treatments compared with 1X (75.0%). Total pigs born was also not affected by treatment, although 2X (11.2) and 3X (10.7) numbers were greater than 1X (10.0). It appears that once-daily estrus detection combined with delayed mating could achieve optimal reproductive performance.

Effects of quality grade, aging period, blade tenderization, and degree of doneness on tenderness of strip loin (2000)

We used 162 strip loins to determine the influence of different quality grades, aging periods, blade tenderization passes, and degree of doneness on thawing and cooking loss and Warner-Bratzler shear force (WBS, tenderness). Select (SEL), Choice (CHO), and Certified Angus Beef™ (CAB) strip loins were aged for 7, 14, or 21 days and not tenderized (0X) or blade tenderized one (1X) or two (2X) times. Steaks from each strip loin were assigned randomly to final endpoint cooking temperatures of 150, 160, and 170°F. For steaks aged 7 days, all quality grade and blade tenderization treatments had similar (P>.05) WBS. For steaks aged 14 days, CHO steaks had lower (P<.05) WBS than SEL steaks, CAB tended (P=.07) to have lower WBS than SEL, 2X steaks had lower (P<.05) WBS than 1X steaks, and 1X steaks had lower (P<.05) WBS than 0X steaks. For steaks aged 21 days, CAB steaks had lower (P<.05) WBS than CHO steaks, CHO steaks had lower (P<.05) WBS than SEL steaks, and 2X steaks had lower (P<.05) WBS than 1X steaks. Among the 0X and 2X groups, CAB and CHO steaks had lower (P<.05) WBS than SEL steaks. For the 1X group, only CAB steaks had lower (P<.05) WBS than SEL steaks. Blade tenderization improved tenderness of strip steaks but should be combined with high quality grades, increased aging, and lower endpoint cooking temperatures to achieve maximum tenderness.

Effects of Training at the Ventilatory Threshold on the Ventilatory Threshold and Performance in Trained Distance Runners

This study compared the effects of different frequencies of training specifically at the ventilatory threshold (VT) in already trained distance runners. The purposes of the investigation were to determine if the training elicited favorable performance enhancements and, if so, to provide information regarding a more beneficial frequency of VT training. Eight trained distance runners underwent maximal testing to determine VO2 peak and VT and completed a timed performance run on the treadmill. Four subjects then added three 20-minute runs per week at the velocity that elicited the VT to their training for 6 weeks (3X), while the other 4 subjects added 1 VT run per week (1X). When retested at 3 weeks, the 3X group exhibited a significant increase in VT, but after 3 additional weeks of training no further significant changes were detected. Conversely, the 1X group showed no alteration in VT after 3 weeks but showed a similar overall increase in VT as the 3X group at the conclusion of the study, despite the much lower frequency of VT training. Both groups also exhibited significant increases in V? O2 peak and an increase in performance of approximately 36%. It was confirmed that training at the VT is an effective method for improving middle distance running performance and that the beneficial changes observed can be achieved just as readily with VT sessions of moderate frequency.