14-d Treatment Research Articles

Controlled atmosphere treatment for control of grape mealybug, Pseudococcus maritimus (Ehrhorn) (Hemiptera: Pseudococcidae), on harvested table grapes

Controlled atmosphere (CA) treatments with ultralow oxygen (ULO) alone and in combinations with 50% carbon dioxide were studied to control grape mealybug, Pseudococcus maritimus (Ehrhorn) on harvested table grapes. Two ultralow oxygen levels, 30 and <0.01μLL−1, were tested in both ULO and ULO+50% CO2 treatments. The ULO treatments with the lower oxygen level were more effective than the ULO treatments at the higher oxygen level. The ULO+50% CO2 treatments were more effective than the ULO treatments. Grape mealybug eggs were significantly more tolerant of ULO and ULO+CO2 treatments than nymphs and adults. A 14day ULO treatment with 30μLL−1 O2 at 2°C did not achieve 100% mortalities of any life stage. In the presence of 50% CO2, the 14d treatment achieved complete mortality of all life stages of the grape mealybug. A 3d ULO treatment with <0.01μLL−1 O2 at 2°C resulted in 93.3% mortality of nymphs and adults. The 3d ULO treatment in combination with 50% CO2 treatments, however, achieved complete control of grape mealybug nymphs and adults and caused 70.5% relative egg mortality. Complete egg mortality was achieved in a 10d ULO+50% CO2 treatment with <0.01μLL−1 O2 at 2°C. Both the 14d CA treatment with 30μLL−1 O2 and 50% CO2 and the 10d CA treatment with <0.01μLL−1 O2 and 50% CO2 were tested on table grapes and grape quality was evaluated after two weeks of post-treatment storage. The CA treatments did not have a significant negative impact on grape quality and were safe for table grapes. The study indicated that CA treatments have potential to be developed for postharvest control of grape mealybug on harvested table grapes.

Phase Ib study of RG7112 with doxorubicin (D) in advanced soft tissue sarcoma (ASTS).

10514 Background: Preclinical data demonstrate p53-dependent MIC-1 activation by both D and the MDM2 inhibitor RG7112. This study evaluated the tolerability of combining D with RG7112 in ASTS patients (pts) and pharmacokinetic (PK) and pharmacodynamic (PD) parameters of the combination. Methods: A phase 1b 3+3 dose escalation study was designed. Pts with ASTS in whom D was considered appropriate were eligible. D was administered IV at either 50 mg/m2 or 60 mg/m2on day 1 with RG7112 administered orally 500 or 1000 mg QD for 3 or 5 days (d1-3 or d1-5) in 28-d treatment cycles. Safety, PK and PD, including serum MIC-1 (an indicator of p53 activation), were assessed. Results: As of January 15, 2013, enrollment was complete with 23 pts with ASTS accrued; safety data for cycle 1 was available for preliminary review in 20 pts. Growth factor support was mandated following the first dosing group (D 60 mg/m2and RG7112 500 mg x 5d) due to febrile neutropenia or grade 4 neutropenia. Of the 20 pts for which cycle 1 safety data is available, 13 pts reported neutropenia (12 grade 3/4); 5 reported grade 3/4 febrile neutropenia (3 following mandatory GCSF prophylaxis); 12 pts developed thrombocytopenia (9 grade 3/4). PK parameters of the combination therapy are similar to single agent values and did not demonstrate evidence of drug-drug interactions. Serum MIC-1 rapidly increased to levels greater than for either agent alone. Best response to date is stable disease in 8/16 pts who have had interim evaluations. Conclusions: Combination therapy with D and RG7112 resulted in a high rate of grade 3/4 neutropenia (60%) or thrombocytopenia (45%). PK analysis does not suggest this is due to changes in the metabolism of either drug. This combination demonstrates apparent potentiation of p53 activation demonstrated by increased MIC-1 levels greater than additive effects of the single agents. Biomarker analyses, safety, PK, and MIC-1 data will be presented. Clinical trial information: NCT01605526. [Table: see text]

Follicular populations and luteal function in dairy heifers treated with a controlled internal drug release insert for 14 days as a method to synchronize the estrous cycle before prostaglandin F2α treatment and artificial insemination

Progesterone-containing controlled internal drug release (CIDR) inserts are used to synchronize the estrous cycle before PGF2α is administered for timed AI (14dCIDR-PGF2α program). The program, initially designed for beef cattle, was recently shown to be efficacious in dairy heifers. We hypothesized that the 14-d CIDR treatment would synchronize the estrous cycle in dairy heifers and result in a uniformly sized corpus luteum (CL) and largest follicle (LF) at the time of PGF2α treatment. Holstein (n=110) or Holstein × Guernsey (n=4) dairy heifers were assigned to 2 treatments: (1) 14dCIDR-PGF2α [CIDR in for 14d, CIDR out for 16d, PGF2α and AI after observed estrus (n=57)] or (2) control [PGF2α and AI after observed estrus (n=57)]. Regardless of treatment, additional PGF2α injections were administered at 14-d intervals to heifers that were not seen in estrus. Ovarian ultrasonography and blood sampling were done on d 0 (CIDR administered), 14 (day CIDR removed), 19 (5d after CIDR removed), 30 (PGF2α administered), and 44 (second PGF2α dose administered to heifers that were not detected in estrus after the first PGF2α). Compared with control (untreated), more CIDR-treated heifers were categorized as having a small CL (≤9.9mm) and large LF (15.0–19.9mm) on d 14 (CIDR removal) and, as expected, a greater percentage of CIDR-treated heifers were in estrus during the 5d after the CIDR removal compared with control heifers (75.4 vs. 22.8%, respectively). On d 19, the CIDR-treated heifers had apparently ovulated based on disappearance of LF and appearance of small CL. On d 30 (PGF2α administration), 89% of 14dCIDR-PGF2α heifers had CL that were ≥20mm in diameter compared with 55% for control. Presence of larger CL on d 30 was associated with greater concentrations of plasma progesterone in 14dCIDR-PGF2α compared with control (10.5±0.5 vs. 5.0±0.6ng/mL, respectively). The percentages of heifers with LF in the smallest category (≤9.9mm) tended to be less (5.3 vs. 16.6%) and the percentage of heifers with LF in the medium-size category (10.0 to 14.9mm) tended to be greater (84.2 vs. 69.1%) for 14dCIDR-PGF2α versus control, respectively, on d 30. More heifers were detected in estrus within 5d after the first PGF2α (86.0 vs. 56.1%) and conception rate to AI using sexed semen tended to be greater (61.2% vs. 40.6%) for 14dCIDR-PGF2α compared with control (respectively). Treating dairy heifers with a CIDR for 14d was an effective method to synchronize an estrous cycle before PGF2α was administered.

Crude glycerol as glycogenic precursor in feed; effects on milk coagulation properties and metabolic profiles of dairy cows

As grain prices rise, the search for alternative glycogenic precursors in animal feed becomes increasingly important, and this study was conducted to determine if the replacement of starch with glycerol, as an alternative glycogenic precursor, affects the milk metabolic profile and milk coagulation ability, and therefore the quality of the milk. Eight primiparous mid-lactation Holstein cows were fed during a replicated 4 × 4 Latin square trial with four different isoenergetic rations: (1) control (T0) fed a total mixed ration (TMR) with barley meal; (2) group T1, decreased barley content, replaced isoenergetically with 1 kg crude glycerol; (3) group T2, the barley meal was replaced with 2 kg of crude glycerol; and (4) group T3 the barley meal was replaced with 3 kg of crude glycerol. Rumen, blood and milk samples were collected at the end of every 21-d treatment period. Rumen samples were analysed for proportion of total volatile fatty acid (VFA), blood samples for insulin and glucose, and milk for metabolites (e.g. citric-acid cycle compounds). The change in glycogenic precursors had a positive effect on rumen VFA proportions; the proportion of propionic acid increased (P < 0.001). Milk protein (P < 0.001) and curd firmness (P < 0.001) both increased. The increase in milk protein concentration may have been due to an increase in microbial protein. Regarding the milk metabolic profiles, different signals were positively associated with coagulation ability and change in the diet. Based on this study, changing the glycogenic precursor in animal diet in this way is possible, and may have no immediate deleterious consequences on milk quality or cow health. Indeed, there is evidence for benefits from this substitution.

In Vivo Blockade of the PD-1 Receptor Suppresses HIV-1 Viral Loads and Improves CD4+ T Cell Levels in Humanized Mice

The programmed death-1 (PD-1) pathway limits the function of virus-specific T cells during chronic infection. We previously showed that blockade of the PD-1 pathway increases HIV-1-associated T cell function in vitro. However, the effect of PD-1 blockade on HIV-1 disease progression in vivo has not been examined. As in humans, HIV-1-infected humanized BALB/c-Rag2(-/-)γc(-/-) (Rag-hu) mice express elevated levels of PD-1 on T cells during chronic infection. To examine the effect of PD-1 blockade on disease progression, Rag-hu mice with chronic HIV-1 infection were treated with a blocking mAb directed against programmed cell death-1 ligand-1, the ligand for PD-1. Programmed cell death-1 ligand-1-treated Rag-hu mice exhibited a progressive decrease in the HIV-1 plasma viral load, with a 7-fold decrease by day 7, a 20-fold decrease by day 14, a 178-fold decrease by day 21, and a 269-fold decrease by day 28 postinitiation of treatment. By day 7, the percentage of CD4(+) T cells was statistically higher in the treated compared with the untreated group, and this trend was sustained throughout the 28-d treatment period. Moreover, there was a strong inverse correlation between plasma viral load and the percentage of both CD4(+) (r = -0.66; p < 0.0001) and CD8(+) (r = -0.64; p < 0.0001) T cells in the treated mice but not the untreated mice. This study provides "proof of concept" that humanized mice can be used to examine the effects of immunotherapeutic interventions on HIV-1 infection. Furthermore, to our knowledge, these data demonstrate for the first time that blockade of the PD-1 pathway reduces HIV-1 viral loads.

Safety and Efficacy of Oral Azacitidine (CC-486) Administered in Extended Treatment Schedules to Patients with Lower-Risk Myelodysplastic Syndromes

AbstractAbstract ▪424▪This icon denotes a clinically relevant abstract Background:Azacitidine for injection has been shown to prolong overall survival in patients (pts) with higher-risk myelodysplastic syndromes (MDS) compared with conventional care regimens (CCR) (Lancet Oncol, 2009). An oral formulation of azacitidine (CC-486) is in development. Oral azacitidine may maximize convenience, eliminate injection-site reactions, and if administered in extended dosing schedules, may enhance and prolong the therapeutic effects of azacitidine. Oral azacitidine administered once-daily (QD) for 7 days (d) of repeated 28d cycles has been shown to be bioavailable, biologically and clinically active, and well-tolerated in pts with MDS and acute myeloid leukemia (Garcia-Manero, J Clin Oncol, 2011). Preliminary evidence suggests that extending oral azacitidine dosing to 14d or 21d of the 28d cycle may enhance pharmacodynamic and epigenetic activity (Laille, Leuk Res, 2011). Purpose:To evaluate hematologic response and safety associated with extended dosing regimens of oral azacitidine in pts with lower-risk MDS. Methods:This ongoing, multicenter, phase 1 study, enrolled pts with lower-risk (IPSS Low or INT-1) MDS who were RBC transfusion dependent (TD) and/or thrombocytopenic (average platelet count ≤50,000 within 56d prior to the first dose) at baseline. Pts were sequentially assigned to receive oral azacitidine 300mg QD for either 14d or 21d of repeated 28d cycles. Hematologic assessments were made every 2 weeks. Hematologic response was assessed using IWG 2006 criteria (Cheson, Blood, 2006). Adverse events (AEs) were graded using NCI-CTCAE version 3.0. Results:At data cut-off (May 18, 2012), 53 pts with lower-risk MDS had enrolled (300mg oral azacitidine QDx14d, n=26; QDx21d, n=27). Demographic and disease characteristics at baseline were similar in the 14d and 21d treatment cohorts (Table 1). Median (range) hematology counts at baseline were Hgb 8.7 g/L (6.0–13.0), ANC 1.6×109/L (0–30.3), and platelets 56.0×109/L (6.0–564.0). At study entry, 40% of pts had received no prior MDS treatment (except transfusions), 45% had received erythropoiesis-stimulating agents, and 15% had received WBC growth factors. The number of oral azacitidine treatment cycles received ranged from 1 to 12 (median numbers of oral azacitidine cycles were 6 in the QDx14d and 4 in the QDx21d cohorts). Four pts in the 21d cohort and 1 pt in the 14d cohort received reduced oral azacitidine doses (200mg QD). Overall, 10 pts discontinued the study, including 6 pts (3 pts in each cohort) who discontinued due to AEs that may have been treatment-related (gastrointestinal [n=2] or intracranial [n=1] hemorrhage, febrile neutropenia [n=1], pneumonia [n=1], thrombocytopenia [n=1]). Overall response rates (ORR), which included complete (CR) and partial remission (PR), any hematologic improvement (HI), and transfusion independence (TI), ranged from 38.5% in the QDx14d cohort to 29.6% in the QDx21d cohort, and RBC TI was achieved by 47% and 33%, respectively, of pts who were RBC TD at baseline (Table 2). For pts who received at least 4 cycles of oral azacitidine (14d, n=19; 21d, n=14), ORR was 47.4% in the 14d and 50.0% in the 21d cohorts, and RBC TI rates in RBC TD pts (n=16) were 67% in the 14d and 57% in the 21d cohorts. The most frequent (≥5%) grade 3/4 hematologic AEs in the QDx14d cohort were anemia (11.5%), thrombocytopenia (11.5%), and neutropenia (7.7%); and in the QDx21d cohort were neutropenia (14.8%), anemia (7.4%), and febrile neutropenia (7.4%). Most frequent grade 3/4 non-hematologic AEs were gastrointestinal, including vomiting (7.7%) in the QDx14d cohort, and diarrhea (11.1%) and vomiting (7.4%) in the QDx21d cohort. Conclusions:Oral azacitidine 300mg QD administered in extended dosing schedules of 14d or 21d of repeated 28d cycles was effective and well-tolerated in these pts with lower-risk MDS. Beside hematologic AEs, the most frequently observed AEs with oral azacitidine were gastrointestinal and were manageable. Efficacy and safety outcomes with 300mg QD oral azacitidine were generally comparable between the 14d and 21d extended dosing regimens. Based on these data, oral azacitidine administered once-daily in extended dosing schedules is active and well-tolerated and warrants further investigation in randomized, controlled trials. [Display omitted] [Display omitted] Disclosures:Garcia-Manero:Celgene: Research Funding, Speakers Bureau. Gore:Celgene Corporation: Consultancy, Research Funding. Scott:Celgene Corporation: Honoraria, Research Funding, Speakers Bureau. Hetzer:Celgene Corporation: Employment, Equity Ownership. Kumar:Celgene Corporation: Employment, Equity Ownership. Skikne:Celgene: Employment, Equity Ownership.

Nutrient demand interacts with grass maturity to affect milk fat concentration and digestion responses in dairy cows

Effects of grass maturity on dry matter intake (DMI), milk production, ruminal fermentation and pool sizes, digestion and passage kinetics, and chewing activity and the relationship of these effects with preliminary DMI (pDMI) were evaluated using 13 ruminally and duodenally cannulated Holstein cows in a crossover design with a 14-d preliminary period and two 18-d treatment periods. During the preliminary period, pDMI of individual cows ranged from 23.5 to 28.2kg/d (mean=26.1kg/d) and 3.5% fat-corrected milk (FCM) yield ranged from 30.8 to 57.2kg/d (mean=43.7kg/d). Experimental treatments were diets containing orchardgrass silage harvested either (1) early-cut, less mature (EC) or (2) late-cut, more mature (LC) as the sole forage. Early- and late-cut orchardgrass contained 44.9 and 54.4% neutral detergent fiber (NDF) and 20.1 and 15.3% crude protein, respectively. Forage:concentrate ratio was 58:42 and 46:54 for EC and LC, respectively; both diets contained approximately 25% forage NDF and 30% total NDF. Preliminary DMI, an index of nutrient demand, was determined during the last 4d of the preliminary period when cows were fed a common diet and used as a covariate. Main effects of grass maturity and their interaction with pDMI were tested by ANOVA. The EC diet decreased milk yield and increased milk fat concentration compared with the LC diet. Grass maturity and its interaction with pDMI did not affect FCM yield, DMI, rumen pH, or microbial efficiency. The EC diet increased rates of ruminal digestion of potentially digestible NDF and passage of indigestible NDF (iNDF) compared with the LC diet. The lower concentration and faster passage rate of iNDF for EC resulted in lower rumen pools of iNDF, total NDF, organic matter, and dry matter for EC than LC. Ruminal passage rates of potentially digestible NDF and starch were related to level of intake (quadratic and linear interactions, respectively) and subsequently affected ruminal digestibility of these nutrients. The EC diet decreased eating, ruminating, and total chewing time per unit of forage NDF intake compared with the LC diet. When grass silage was the only source of forage in the diet, cows supplemented with additional concentrate to account for decreasing protein and increasing fiber concentrations associated with more mature grass had similar feed intake and produced similar FCM yields as cows fed less mature grass.

Short communication: Presynchronization for timed artificial insemination in grazing dairy cows by using progesterone for 14 days with or without prostaglandin F2α at the time of progesterone withdrawal

Progesterone-containing devices can be inserted intravaginally for 14 d to presynchronize the estrous cycle for timed artificial insemination (TAI) in beef heifers (“14-day CIDR-PG” or “Show-Me-Synch” program). The progesterone treatment is effective for presynchronization because cattle develop a persistent dominant follicle during treatment that ovulates within 3 d after progesterone removal. The subsequent estrous cycle can be effectively used for a TAI program. Some cattle will retain a functional corpus luteum (CL) for the entire 14-d treatment period and will not be synchronized effectively because the interval to ovulation depends on the lifespan of their existing CL. The objective was to test the effect of a luteolytic dose of PGF2α at progesterone removal for improving synchrony of estrus after treatment and increasing conception rate to a subsequent TAI in dairy cows. Postpartum cows (n = 1,021) from 2 grazing dairy herds were assigned to 1 of 2 presynchronization programs that used a controlled internal drug releasing (CIDR) device containing progesterone: 14dCIDR (CIDR in, 14 d, CIDR out; n = 523) or 14dCIDR+PGF2α (CIDR in, 14 d, CIDR out, and PGF2α; n = 498). Cows were body condition scored (BCS; 1 to 5, thin to fat) and tail painted at CIDR removal. Paint score (PS) was recorded after CIDR removal [PS = 0 (all paint removed, indication of estrus), PS = 3 (paint partially removed), or PS = 5 (no paint removed; indication of no estrus)]. At 19 d after CIDR removal, all cows were treated with PGF2α, 56 h later treated with GnRH, and then 16 h later were TAI. Treating cows with PGF2α at CIDR removal increased the percentage with PS = 0 within 5 d (58.1% vs. 68.9%; 14dCIDR vs. 14dCIDR+PGF2α). We found no effect of treatment, however, on conception rate at TAI (41.1% vs. 43.6%; respectively). The TAI conception rate increased with increasing BCS and was greater for cows that had PS = 0 within 5 d after CIDR removal. In summary, treating cows with PGF2α at CIDR removal increased the percentage of cows with all tail paint removed but did not increase percentage of pregnant cows after TAI.

The effects of overstocking Holstein dairy cattle during the dry period on cortisol secretion and energy metabolism

The objective was to determine whether overstocking during the dry period could alter physiological parameters in dairy cattle associated with cortisol secretion and energy metabolism. Four groups of 10 late-gestation, nonlactating Holstein cows (6 multiparous cows and 4 heifers per group) were exposed to both a control [1 lying stall/cow and 0.67m of linear feed bunk (FB) space/cow] and an overstocked (1 stall/2 cows and 0.34m of FB space/cow) stocking density treatment in a replicated crossover design with 14-d treatment periods. On d 1, 3, 5, 7, 9, and 11 of each 14-d treatment period, blood and fecal samples were collected from each cow for the determination of plasma nonesterified fatty acids (NEFA), glucose, insulin, and fecal cortisol metabolite (11,17-dioxoandrostane; 11,17-DOA) concentrations. Glucose and ACTH challenges were conducted on d 13 and 14, respectively, of each treatment period. Dry matter intake per cow was greater during the overstocked period than during the control period (15.9 vs. 14.9±0.5kg/d). Plasma NEFA and glucose concentrations were greater (0.11 vs. 0.09±0.006mEq/L and 65.3 vs. 64.2±1.1mg/dL, respectively) and 11,17-DOA concentration tended to be greater (891 vs. 792±86ng/g of fecal dry matter) during the overstocked period than during the control period. Insulin concentration was the same during the overstocked (29.0±2.1μIU/mL) and control (31.2±2.1μIU/mL) periods. Overstocking was associated with slightly slower glucose clearance from circulation as evidenced by a greater area under the curve estimate for the glucose response curves (2,882 vs. 2,657±165mg/dL × 180min) but a more attenuated insulin response (insulin area under the curve = 5,258 vs. 6,692±1,104μIU/mL × 180min for the overstocked and control periods, respectively). Changes in tissue glucose uptake may be mediated by changes in pancreatic insulin secretion or peripheral tissue responses to insulin. The role of glucocorticoids in mediating these changes in energy metabolism is still unclear because stocking density treatment was not associated with changes in adrenal secretion of cortisol following ACTH stimulation.

Nutrient demand interacts with forage family to affect digestion responses in dairy cows

Effects of forage family on dry matter intake (DMI), milk production, ruminal pool sizes, digestion and passage kinetics, and chewing activity and the relationship of these effects with preliminary DMI (pDMI), an index of nutrient demand, were evaluated using 13 ruminally and duodenally cannulated Holstein cows in a crossover design with a 14-d preliminary period and two 18-d treatment periods. During the preliminary period, pDMI of individual cows ranged from 19.6 to 29.5kg/d (mean=25.9kg/d) and 3.5% fat-corrected milk yield ranged from 24.3 to 60.3kg/d (mean=42.1kg/d). Experimental treatments were diets containing either a) alfalfa silage (AL) or b) orchardgrass silage (OG) as the sole forage. Alfalfa and orchardgrass contained 42.3 and 58.2% neutral detergent fiber (NDF) and 22.5 and 11.4% crude protein, respectively. Forage:concentrate ratios were 60:40 and 43:57 for AL and OG, respectively; both diets contained approximately 25% forage NDF and 30% total NDF. Preliminary DMI was determined during the last 4 d of the preliminary period when cows were fed a common diet and used as a covariate. Main effects of forage family and their interaction with pDMI were tested by ANOVA. Forage family and its interaction with pDMI did not affect feed intake, milk yield, or milk composition. The AL diet increased indigestible NDF (iNDF) intake and decreased potentially digestible NDF (pdNDF) intake compared with OG. The AL diet increased ruminal pH, digestion rates of pdNDF and starch, and passage rates of pdNDF and iNDF compared with OG, which affected ruminal digestibility. Passage rate of iNDF was related to pDMI; AL increased iNDF passage rate and OG decreased it as pDMI increased. The AL diet decreased ruminal pool sizes of pdNDF, starch, organic matter, dry matter, and rumen digesta wet weight and volume compared with OG. The AL diet decreased ruminating time per unit of forage NDF consumed compared with OG, indicating that alfalfa provided less physically effective fiber than orchardgrass. The AL diet, but not OG, increased ammonia N, nonammonia nonmicrobial N, and nonammonia N fluxes as pDMI increased. Efficiency of microbial protein synthesis was positively related to pdNDF passage rate for OG, but not AL. The faster rates of digestion and passage for AL compared with OG decreased rumen pool size but did not increase feed intake for cows consuming AL. Digestion responses to forage family were affected by nutrient demand of cows.

Rates of particle size reduction and passage are faster for legume compared with cool-season grass, resulting in lower rumen fill and less effective fiber

Effects of forage family on rates of particle size reduction in, and passage from, the rumen and the relationship of these effects with preliminary dry matter intake (pDMI) were evaluated using 13 ruminally and duodenally cannulated Holstein cows in a crossover design with a 14-d preliminary period and two 18-d treatment periods. During the preliminary period, pDMI of individual cows ranged from 19.6 to 29.5kg/d (mean = 25.9kg/d). Experimental treatments were diets containing either a) alfalfa silage (AL) or b) orchardgrass silage (OG) as the sole forage. Silages were chopped to 10-mm theoretical length of cut and contained 42.3 and 58.2% neutral detergent fiber (NDF) for alfalfa and orchardgrass, respectively. Both diets contained approximately 25% forage NDF and 30% total NDF. Feed, orts, rumen, and duodenal samples were wet sieved to fractionate particles above (large) and below (small) 2.36mm. Indigestible NDF (iNDF) was used as a flow marker. Preliminary DMI, an index of nutrient demand, was determined during the last 4 d of the preliminary period when cows were fed a common diet and used as a covariate. Main effects of forage family and their interaction with pDMI were tested by ANOVA. Approximately 75% of the NDF consumed was large and 25% was small for both treatments, but cows fed AL consumed more iNDF and less potentially digestible NDF (pdNDF) than cows fed OG. The AL diet increased the reduction rate (large to small) compared with OG despite less rumination per unit of forage NDF for AL than OG, suggesting alfalfa NDF was more fragile than orchardgrass NDF. Over 55% of particles in the rumen were below 2.36mm for AL and OG, indicating that particle size was not a limiting constraint to passage. Passage rates (kp) of large iNDF and large pdNDF were similar for AL and OG, but AL increased kp of large pdNDF and OG decreased it as pDMI increased. The AL diet increased kp of small iNDF and small pdNDF compared with OG, resulting in lower rumen fill for AL than OG. The kp of small iNDF and small pdNDF were similar within forage family, suggesting buoyancy was not limiting passage. The OG diet increased rumen pool size of large NDF compared with AL, which likely retained small NDF, contributing to the slower kp of small iNDF and small pdNDF observed for OG. Particle size reduction was a prerequisite to ruminal passage but not a constraint. Selective retention of small particles was less for alfalfa than orchardgrass, resulting in lower rumen fill and less effective fiber.

Nutrient demand interacts with grass particle length to affect digestion responses and chewing activity in dairy cows

Effects of grass particle length on dry matter intake (DMI), milk production, ruminal fermentation and pool sizes, digestion and passage kinetics, and chewing activity and the relationship of these effects with preliminary DMI (pDMI) were evaluated using 15 ruminally and duodenally cannulated Holstein cows in a crossover design with a 14-d preliminary period and two 18-d treatment periods. During the preliminary period, pDMI of individual cows ranged from 22.6 to 29.8kg/d (mean=25.8kg/d) and 3.5% fat-corrected milk yield ranged from 29.2 to 56.9kg/d (mean=41.9kg/d). Experimental treatments were diets containing orchardgrass silage chopped to either (a) 19-mm (long) or (b) 10-mm (short) theoretical length of cut as the sole forage. Grass silages contained approximately 46% neutral detergent fiber (NDF); diets contained 50% forage, 23% forage NDF, and 28% total NDF. Preliminary DMI, an index of nutrient demand, was determined during the last 4 d of the preliminary period when cows were fed a common diet and used as a covariate. Main effects of grass particle length and their interaction with pDMI were tested by ANOVA. Grass particle length and its interaction with pDMI did not affect milk yield, milk composition, or rumen pH. Long particle length tended to decrease DMI compared with short particle length, which might have been limited by rumen fill or chewing time, or both. Passage rates of feed fractions did not differ between long and short particle lengths and were not related to level of intake. As pDMI increased, long particles decreased ruminal digestion rate of potentially digestible NDF at a faster rate than short particles. As a result, long particles decreased or tended to decrease rates of ruminal turnover for NDF, organic matter, and dry matter and increased their rumen pools compared with short particles for cows with high pDMI. Long particles increased eating time, which affected cows with high intake to the greatest extent, and total chewing time compared with short particles. As intake increased, ruminal digestion (kg/d) and digestibility (%) of starch decreased, rumen pool size of starch increased, and postruminal digestion and digestibility of starch increased quadratically. When grass silage was the only source of forage in the diet, increasing chop length from 10 to 19mm tended to decrease DMI but did not negatively affect productivity of cows, which were fed adequate fiber.

Efficacy of quinine against ichthyophthiriasis in common carp Cyprinus carpio

Ichthyophthiriasis, caused by Ichthyophthirius multifiliis, is an economically important worldwide parasitic disease that infects all freshwater fish. Since the banning of malachite green for use in food fish, there has been a great need for alternative therapeutants. The goal of this study was to investigate the efficacy of quinine against I. multifiliis. Parasite developmental stages from our laboratory-established life cycle in rainbow trout Oncorhynchus mykiss were exposed to quinine in vitro, and a dual fluorescent staining technique was used to allow a clear distinction between viable and damaged parasites. Furthermore, the effect of quinine was assessed in vivo by oral administration and intraperitoneal injections in common carp Cyprinus carpio. The results of the in vitro experiments proved quinine to be effective against the parasite. Quinine injected at a dosage of 60 mg kg(-1) body weight resulted in a significantly lower number of trophonts. In contrast, in-feed trials did not show a significant reduction of trophonts after treatment commencing 1 d after infection with concentrations of up to 20 g quinine kg(-1) feed for 3 d. After a 14-d treatment at concentrations of up to 10 g quinine kg(-1) feed prior to theront exposure, there was also no significant difference in parasite numbers between treated and control groups. The results of oral versus parenteral application of quinine indicate that the substance is not completely absorbed from the intestinal tract of common carp. However, medicated feed containing higher concentrations of quinine was less readily accepted by the fish, presumably due to the bitter taste.

Targeting PIM kinase activity significantly augments the efficacy of cytarabine

Targeting PIM kinase activity significantly augments the efficacy of cytarabine

Comparative Germination and Emergence Ecology of Two Populations of Common Lambsquarters (Chenopodium album) from Iran and Denmark

Common lambsquarters, a summer annual weed, has occupied comparatively different ecogeographical regions around the globe. To investigate the extent to which germination and emergence traits have differentiated in two natural populations of common lambsquarters from different environments (Denmark as mesic and Iran as xeric population), experiments were conducted in the laboratory. Germination of both populations was stimulated by light. The greatest germination percentage of xeric and mesic populations occurred at 25/15 and 20/10 C, respectively. The xeric population showed significantly greater germinability at the lowest (15/5 C) and the highest (35/25 C) fluctuating temperature regimes. Germination of the xeric population was only slightly affected, and the mesic population was severely retarded at a salinity level of 20 desi siemens (DS) m−1. Greater than 42% of the xeric seeds germinated at 30 dS m−1salinity, while the mesic germination almost ceased at this salinity level. Germination of seeds placed in distilled water after the 14-d salinity exposure treatments (recovery rates) was also greater for the xeric vs. mesic populations. Xeric populations showed more than 65% germination up to the osmotic potential of −0.4 MPa, while decreasing osmotic potential from 0 to −0.4 MPa caused an 80% reduction in mesic population germinability (9% germination). The greatest emergence of xeric (77%) and mesic (70%) populations occurred for seeds placed on the soil surface and no seedlings emerged from burial depth 3 cm. These study results suggest that common lambsquarters populations have differentiated for heat, salinity, and drought tolerance at germination stage. These contrasting germination patterns are appeared to be due to either adaptation via natural selection or maternal effects, or some combination of both. Though this study has certain limitations, it, through its findings and their propositions, adds to the existing knowledge about interpopulation differences in germination requirements of common lambsquarters, as a globally distributed species.

Polychlorinated biphenyls-containing electrical insulating oil contaminated soil treatment with calcium and magnesium peroxides

Calcium and magnesium peroxides were applied for the treatment of soil contaminated by polychlorinated biphenyls-containing electrical insulating oil (Aroclor 1016). The removal of PCB-containing electrical insulating oil was achieved with the addition of either calcium peroxide or magnesium peroxide alone and dependent on dosages of the chemical. A 21-d treatment of 60% watered soil with the moderate addition (chemical/oil weight ratio of 0.005/1) of either calcium peroxide or magnesium peroxide resulted in nearly complete (96 ± 2%) oil removal, unsubstantial increase in soil pH and almost no changes in oxygen consumption and dehydrogenase activity, making it suitable for the soil decontamination.

N-Methyl-2-Pyrrolidone Is a Biologically Active Thalidomide Subunit with In Vivoanti-Myeloma Properties

AbstractAbstract 4082N-methyl-2-pyrrolidone (NMP) is considered an inert drug delivery vehicle that is widely used as an industrial organic solvent. NMP is found in a range of pharmaceutical preparations and bio-prosthetic cements. In particular, NMP is used to solubilize kinase inhibitors for pre-clinical testing in experimental mice.The Vk*MYC transgenic mouse is a model of multiple myeloma (MM) that recapitulates the human disease. We have adapted Vk*MYC MM using a syngeneic transplant approach to shorten latency and facilitate testing of novel therapeutics. In this model, we noted delayed disease progression and improved survival in NMP treated mice compared to alternative vehicle controls. To confirm this observation, matched cohorts of mice bearing monoclonal kappa-secreting MM were treated with 20% NMP/80% polyethylene glycol (PEG) (n = 13), PEG alone (n = 10) or PBS (n = 7) at 100uL per 20g mouse daily p.o. Mice were monitored by serial serum protein electrophoresis and quantitative kappa immunoglobulin (IgK) estimation. At the commencement of therapy, cohorts were well matched for disease burden; mean M-spikes 12%, 14% and 14% of total serum protein for NMP, PBS and PEG respectively (p=0.58 NMP vs. PBS, p=0.68 NMP vs. PEG). After 28d treatment, NMP-treated mice had lower paraproteinemia compared to PBS and PEG controls (mean M-spike NMP 31%, PBS 42% and PEG 40%; p < 0.05 NMP vs. PBS or PEG), with concordant suppression of IgK (NMP 18g/L vs. PBS 39g/L, p<0.05). Survival in NMP-treated mice was also significantly longer than that in PBS-treated controls (median survival 50d and 40d respectively; p<0.05 log rank). Survival of PEG-only treated mice did not differ from PBS controls (median survival 39.5d; p=0.49).We compared the structure of NMP to known MM therapeutics, revealing that NMP is a subunit of thalidomide and its analogues. We next tested NMP for cytotoxicity and immune-modulatory drug (IMiD) activity in vitro. NMP (1uM – 10mM) was not directly cytotoxic to healthy donor peripheral blood mononuclear cells (PBMCs) or the U266 myeloma cell line. However, treatment of PBMC with 10uM NMP significantly enhanced NK cell cytotoxicity against the NK cell-sensitive target cell line K562 from 26% to 42% (p < 0.05). Further, NMP favored a Th-1 cytokine profile without directly altering the frequency of CD4, CD8, NK or regulatory T cell subsets in vitro. As compared to the IMiD lenalidomide, NMP inhibited TNF-a production from LPS stimulated monocytes. Given the IMiD-like effects of NMP, we assessed its ability to alter the expression of NK cell ligands on myeloma targets. NMP upregulated both NKG2D and DNAM-1 ligands (MIC-A, MIC-B, ULBP-1, ULBP-2 and CD155) on U266 cells, thereby enhancing the NK cell mediated lysis of the U266 cell line.Our results demonstrate the in vivo anti-myeloma activity and IMiD-like properties of NMP, indicating that it is a putative active thalidomide subunit. The detection of IMiD-like activity was facilitated by a sensitive, immune competent MM model. NMP should no longer be considered inert for the purposes of drug delivery. These findings mandate re-evaluation of drug efficacy data where NMP has been used as the drug delivery vehicle, particularly in immune-competent cancer models. Our findings with NMP may allow the development of new IMiDs. Finally, occupational or iatrogenic exposure to NMP should be further assessed as a potential risk for immunologically mediated adverse effects. Disclosures:No relevant conflicts of interest to declare.

Capecitabine for skin cancer prevention in solid organ transplant recipients

Skin cancers are the most common malignancies in solid organ transplant recipients (SOTR). A case-observational, retrospective study was performed to determine the efficacy of low-dose capecitabine in the secondary prevention of skin cancers in SOTRs treated at a single institution. SOTRs with recurrent squamous cell carcinoma (SCC) and/or basal cell carcinoma (BCC) were given low-dose capecitabine 1 g/m(2) daily, days 1-14 of a 21-d treatment cycle. Skin surveillance was performed by dermatologists every 1-3 months. Cumulative incidence rates of SCC, BCC, and actinic keratosis (AK) before and after treatment were scored and statistically compared for each patient using a non-parametric Wilcoxon signed rank test. Fifteen patients (13 men and two women) with a median age of 57 yr (range 40-73) were treated. Incidence rates as measured by mean number of events per month declined by 0.33 for SCC, 0.04 for BCC, and 2.45 for AK (p < 0.05). The most common grade 3 and 4 toxicities included fatigue (40.0%), hand-foot syndrome (20.0%), and diarrhea (20.0%). The discontinuation rate at one yr was approximately 33.3%. We conclude that oral capecitabine significantly decreases the incidence rates of recurrent SCC, BCC, and AK in SOTRs and is associated with manageable toxicity.

Effect of water addition to a total mixed ration on feed temperature, feed intake, sorting behavior, and milk production of dairy cows

The objective of this study was to determine the effects of water addition to a high-moisture total mixed ration (TMR) on feed temperature, feed intake, feed sorting behavior, and milk production of dairy cows. Twelve lactating Holstein cows (155.8±60.1 DIM), individually fed once daily at 1000h, were exposed to 3 diets in a Latin square design with 28-d treatment periods. Diets had the same ingredient composition [30.9% corn silage, 30.3% alfalfa haylage, 21.2% high-moisture corn, and 17.6% protein supplement; dry matter (DM) basis] and differed only in DM concentration, which was reduced by the addition of water. Treatment diets averaged 56.3, 50.8, and 44.1% DM. The study was conducted between May and August when environmental temperature was 18.2±3.6°C and ambient temperature in the barn was 24.4±3.3°C. Dry matter intake (DMI) was monitored for each animal for the last 14 d of each treatment period. For the final 7 d of each period, milk production was monitored, feed temperature and ambient temperature and humidity were recorded (daily at 1000, 1300, and 1600h), and fresh feed and orts were sampled for determination of sorting. For the final 4 d of each period, milk samples were taken for composition analysis. Samples taken for determining sorting were separated using a Penn State Particle Separator that had 3 screens (19, 8, and 1.18mm) and a bottom pan, resulting in 4 fractions (long, medium, short, and fine). Sorting was calculated as the actual intake of each particle size fraction expressed as a percentage of the predicted intake of that fraction. Greater amounts of water added to the TMR resulted in greater increases in feed temperature in the hours after feed delivery, greater sorting against long particles, and decreased DMI, reducing the overall intake of starch and neutral detergent fiber. Milk production and composition were not affected by the addition of water to the TMR. Efficiency of production of milk was, however, increased with greater amounts of water added to the TMR. The increases in feed temperature in the hours after feed delivery were enhanced by higher ambient temperatures; this may be indicative of feed spoilage and thus may have contributed to the reduced DMI observed. Overall, these results suggest that the addition of water to high-moisture TMR (less than 60% DM) containing primarily haylage and silage forage sources will not always discourage cows from sorting, but rather may increase this behavior and limit the nutrient consumption of cows, particularly when ambient temperature is high.

Effects of corticosterone treatment in laying Japanese quail

1. The effects of exogenous corticosterone on plasma corticosterone concentrations, body weight, food intake, the percentage of birds that laid an egg each day and egg weight were measured in laying Japanese quail. Data were collected from birds in the week before corticosterone treatment began, during a 21-d treatment period, and in the 22 d after treatment ended. 2. Groups of quail were treated with corticosterone at three different concentrations in their drinking water. Corticosterone intakes were calculated for each bird and birds were then assigned to 4 intake groups. The groups were 0 (control group), 0·31–0·60, 0·61–0·90, 0·91–1·50 or >1·51 mg corticosterone/bird/d. 3. Plasma corticosterone concentrations in the 4 intake groups increased from around 1 ng/ml on the day before corticosterone treatment began to maximum mean concentrations of 13–18 ng/ml on day 14 of treatment. Mean corticosterone concentrations did not change between day 14 of treatment and the day after treatment ended, and had decreased in only one of 4 intake groups one week later. 4. Mean body weight in the highest intake group remained significantly lower than in controls 22 d after corticosterone treatment ended. Whilst there was no clear effect of corticosterone on food intake during treatment, mean food intake in the three highest corticosterone intake groups was significantly lower than in controls in the week after treatment ended. The percentage of birds that laid an egg each day and egg weight were both decreased by corticosterone, and the percentage of birds that laid an egg each day remained significantly lower in the highest corticosterone intake group compared with controls in the third week after treatment ended. 5. It is suggested that elevated plasma corticosterone concentrations in quail after treatment ended were maintained by a hyperactive hypothalamo–pituitary–adrenal axis for at least one week. Although none of the negative effects of corticosterone were evident in the group of quail with the lowest corticosterone intakes, the findings of the present study show that corticosterone treatment can affect birds for up to several weeks after corticosterone intake ends.