Impact of the induction phase chemotherapy on cytokines and oxidative markers in peripheral and bone marrow plasma of children with acute lymphocytic leukemia.

B-cell acute lymphocytic leukemia (B-ALL) is the main neoplasia affecting children worldwide, in which cytotoxic chemotherapy remains the main treatment modality. In this study, we analyzed the profile of inflammatory markers concerning oxidative stress and cytokines in 17 B-ALL patients. Peripheral blood (PB) and bone marrow (BM) samples were collected and evaluated for the pro-oxidative status (nitric oxide products-NOx and hydroperoxides), antioxidants (sulfhydryl groups-SH and total radical-trapping antioxidant parameter-TRAP), and cytokines (TNF-α, IFN-γ), at diagnosis (D0) to and the end of the induction phase (D28). At D28, hydroperoxides were higher in PB, concomitant to TNF-α levels. INF-γ was increased in the BM at D28. Hydroperoxides were higher in patients presenting malignant cells in BM and/or PB after treatment, a condition named minimal residual disease (MRD) when compared to those without MRD at D28. These findings suggest that oxidative stress and cytokines vary across the B-ALL induction phase, and lipid peroxidation is a potential marker associated with MRD status.

Hepatic mRNA expression for genes related to somatotropic axis, glucose and lipid metabolisms, and inflammatory response of periparturient dairy cows treated with recombinant bovine somatotropin.

Objectives of this experiment were to evaluate the effects of recombinant bovine somatotropin (rbST) treatment of periparturient dairy cows on hepatic mRNA expression for genes related to the somatotropic axis, insulin, glucose, and lipid metabolism, inflammation, and oxidative stress. Holstein cows were enrolled in the experiment at 253 ± 3 d of gestation and assigned to 1 of 3 treatments: untreated control (n = 53), 87.5 mg of rbST (n = 56; rbST87.5), and 125 mg of rbST (n = 57; rbST125). Cows in the rbST87.5 and rbST125 treatments received weekly injections of rbST from -21 to 28 d relative to calving. A subsample of cows (control = 20, rbST87.5 = 20, rbST125 = 20) was randomly selected for collection of liver samples according to expected calving date, BCS, and previous lactation 305-d mature equivalent milk yield. Only cows that had liver sampled at -21 ± 3, -7 ± 3, and 7 ± 3 d relative to calving were used in the current experiment. Blood, sampled weekly from -28 to 21 d relative to calving, was used to determine the concentrations of growth hormone, insulin-like growth factor 1, insulin, cortisol, fatty acids, ß-hydroxybutyrate, glucose, haptoglobin, and tumor necrosis factor-α. Liver samples were used to determine hepatic mRNA expression of 50 genes. Treatment with rbST increased growth hormone concentrations during the postpartum period (control = 9.0 ± 0.7, rbST87.5 = 15.3 ± 1.0, rbST125 = 18.5 ± 1.3 ng/mL) and increased insulin-like growth factor 1 concentrations during the prepartum period (control = 107.4 ± 7.2, rbST87.5 = 126.9 ± 6.6, rbST125 = 139.4 ± 6.9 ng/mL). Control cows had greater postpartum concentrations of ß-hydroxybutyrate (control = 776.4 ± 64.0, rbST87.5 = 628.4 ± 59.7, rbST125 = 595.4 ± 60.9 µmol/L) than rbST cows. The rbST87.5 and rbST125 treatments upregulated the hepatic mRNA expression for somatotropic axis genes (GHR, GHR1A, IGF1, IGFBP3, and SOCS2) on d -7 relative to calving and upregulated the mRNA expression for SOCS2 on d 7. On d -7, rbST87.5 and rbST125 treatments increased mRNA expression for genes involved in hepatic lipid transport (ANGPTL4, APOA5, APOB100, and SCARB1) and downregulated mRNA expression for PPARD, which is involved in lipid storage. On d 7, rbST tended to upregulate the mRNA expression for genes involved in gluconeogenesis (PCK1) and fatty acid ß-oxidation (ACOX1), and downregulated the mRNA expression for genes involved in inflammation (TNFRSF1A, ICAM1, CXCL1, MYD88, HIF1A, IL1RN, NFKBIA, and SOCS3) and oxidative stress (XBP1). Administration of rbST during the periparturient period may improve liver function and health by increasing hepatic capacity for gluconeogenesis and lipid transport and by reducing inflammation and oxidative stress.

Effects of recombinant bovine somatotropin during the periparturient period on innate and adaptive immune responses, systemic inflammation, and metabolism of dairy cows.

The aim of this experiment was to determine effects of treating peripartum dairy cows with body condition score ≥3.75 with recombinant bovine somatotropin (rbST) on immune, inflammatory, and metabolic responses. Holstein cows (253±1d of gestation) were assigned randomly to 1 of 3 treatments: untreated control (n=53), rbST87.5 (n=56; 87.5mg of rbST), and rbST125 (n=57; 125mg of rbST). Cows in the rbST87.5 and rbST125 treatments received rbST weekly from -21 to 28d relative to calving. Growth hormone, insulin-like growth factor 1, haptoglobin, tumor necrosis factor α, nonesterified fatty acids, ß-hydroxybutyrate, glucose, and cortisol concentrations were determined weekly from -21 to 21d relative to calving. Blood sampled weekly from -14 to 21d relative to calving was used for hemogram and polymorphonuclear leukocyte (PMNL) expression of adhesion molecules, phagocytosis, and oxidative burst. Cows were vaccinated with ovalbumin at -21, -7, and 7d relative to calving, and blood was collected weekly from -21 to 21d relative to calving to determine IgG anti-ovalbumin concentrations. A subsample of cows had liver biopsied -21, -7, and 7d relative to calving to determine total lipids, triglycerides, and glycogen content. Growth hormone concentrations prepartum (control=11.0±1.2, rbST87.5=14.1±1.2, rbST125=15.1±1.3ng/mL) and postpartum (control=14.4±1.1, rbST87.5=17.8±1.2, rbST125=21.8±1.1ng/mL) were highest for rbST125 cows. Cows treated with rbST had higher insulin-like growth factor 1 concentrations than control cows (control=110.5±4.5, rbST87.5=126.2±4.5, rbST125=127.2±4.5ng/mL) only prepartum. Intensity of L-selectin expression was higher for rbST125 than for control and rbST87.5 cows [control=3,590±270, rbST87.5=3,279±271, rbST125=4,371±279 geometric mean fluorescence intensity (GMFI)] in the prepartum period. The PMNL intensities of phagocytosis (control=3,131±130, rbST87.5=3,391±133, rbST125=3,673±137 GMFI) and oxidative burst (control=9,588±746, rbST87.5=11,238±761, rbST125=12,724±781 GMFI) were higher for rbST125 cows than for control cows during the prepartum period. Concentrations of serum IgG anti-ovalbumin tended to be higher for rbST125 cows than for control cows (control=0.75±0.11, rbST87.5=0.94±0.10, rbST125=1.11±0.11 optical density) in the prepartum period. Haptoglobin concentration was significantly reduced 7d postpartum for rbST125 treatment compared with control and rbST87.5 treatments (control=2.74±0.28, rbST87.5=2.81±0.28, rbST125=1.87±0.28 optical density). Although treatment tended to affect postpartum ß-hydroxybutyrate (control=747.5±40.2, rbST87.5=753.2±40.1, rbST125=648.8±39.7 µmol/L), it did not affect liver contents of total lipids, triglycerides, or glycogen. Incidence of metritis among rbST125 cows was reduced compared with that in control cows (control=23.1, rbST87.5=18.0, rbST125=7.8%). Treatment of dairy cows with 125mg of rbST improved innate immune responses and IgG concentration, with limited effects on metabolism.

Prepartum stocking density: effects on metabolic, health, reproductive, and productive responses.

The objectives of the current experiment were to determine the effects of 2 prepartum stocking densities on milk yield, concentration of metabolites during the peripartum period, and health and reproductive parameters of dairy cows. Jersey cows enrolled in the experiment at 254±3 d of gestation were balanced for parity (nulliparous vs. parous) and previous lactation projected 305-d mature equivalent milk yield (parous) and assigned to 1 of 2 treatments: 80% headlock stocking density (80SD; 38 animals/48 headlocks) and 100% headlock stocking density (100SD; 48 animals/48 headlocks). The number of experimental units was 8 (4 replicates and 2 pens/treatment per replicate). In total, 154 nulliparous and 184 parous animals were enrolled in the 80SD treatment and 186 nulliparous and 232 parous animals were enrolled in the 100SD treatment. At the start of each replicate, treatments were switched within pen. Cows were milked thrice daily and monthly milk yield, fat and protein content, and somatic cell count data were recorded up to 155 d postpartum. Plasma nonesterified fatty acid concentration was measured weekly, from -18±3 to 17±3 d relative to calving, and plasma ß-hydroxybutyrate was measured weekly, from 1±2 to 17±3 d relative to calving. Cows were examined 1, 4±1, 7±1, 10±1, and 13±1 d relative to calving for diagnosis of uterine diseases. Blood was sampled for determination of progesterone concentration and resumption of ovarian cycles 35±3 and 45±3 d relative to calving. Average headlock (74.1±0.4 vs. 94.5±0.3%) and stall (80.8±0.4 vs. 103.1±0.4%) stocking density was lower for the 80SD treatment compared with the 100SD treatment. Treatment did not affect incidence of retained fetal membranes (80SD=5.1, 100SD=7.8%), metritis (80SD=21.2, 100SD=16.7%), acute metritis (80SD=9.9, 100SD=9.4%), and vaginal purulent discharge (80SD=5.8, 100SD=7.9%). Concentrations of nonesterified fatty acids (80SD=251.5±6.1, 100SD=245.9±5.6µmol/L) and ß-hydroxybutyrate (80SD=508.2±14.3, 100SD=490.9±13.6µmol/L) were not different between treatments. Treatment had no effect on percentage of cows removed from the herd on the first 60 d postpartum (80SD=6.1, 100SD=5.1%) and on rate of removal from the herd up to 305 d postpartum 80SD=referent, 100SD [adjusted hazard ratio (95% confidence interval)]=1.02 (0.75, 1.38). Percentages of cows pregnant to first (80SD=41.9, 100SD=48.4%) and second (80SD=49.3, 100SD=42.0%) postpartum AI were not different between treatments. Finally, treatment did not affect energy-corrected milk yield up to 155 d postpartum (80SD=33.8±0.5, 100SD=33.4±0.5kg/d). In herds with weekly or twice weekly movement of new cows to the prepartum pen and separate housing of nulliparous and parous animals, a target stocking density of 100% of headlocks on the day of movement is not expected to affect health, metabolic, reproductive, and productive parameters.

Effect of an Ovsynch56 protocol initiated at different intervals after insemination with or without a presynchronizing injection of gonadotropin-releasing hormone on fertility in lactating dairy cows.

The objectives of this study were to evaluate effects of 2 resynchronization protocols beginning at different intervals after artificial insemination (AI) on the pattern of return to estrus, ovarian responses, and pregnancy per AI (P/AI) to reinsemination. Lactating cows from 2 dairies, located in Texas (n=2,233) and Minnesota (n=3,077), were assigned to 1 of 4 timed AI (TAI) protocols 17 ± 3 d after AI. All cows were examined for pregnancy 31 ± 3 d after previous AI. Cows assigned to early Ovsynch56 (E-OV56) or OV56 received the Ovsynch56 protocol starting 24 or 31 d after AI, respectively. Cows assigned to early GnRH-GnRH-PGF(2α)-GnRH (E-GGPG) or GGPG received a presynchronizing GnRH injection 17 or 24 d after AI, respectively, 7 d before the start of the Ovsynch56 protocol. Cows observed in estrus after enrollment were inseminated on the same day. Ovaries were examined and blood was sampled for progesterone concentration on the day of first GnRH and PGF(2α) injection of the Ovsynch56 protocol. Pregnancy was diagnosed at 31 and 66 d after resynchronized AI. On the day of the first GnRH injection of the TAI, a higher percentage of cows on E-GGPG and GGPG protocols had a corpus luteum (E-GGPG=83.8, GGPG=91.2, E-OV56=80.4, and OV56=75.5%) and progesterone concentration >1 ng/mL (E-GGPG=62.5, GGPG=76.0, E-OV56=53.6, and OV56=60.8%) than cows assigned to other protocols. However, the percentage of cows ovulating to the first GnRH injection of TAI was not affected by treatment. Fewer E-GGPG and more OV56 cows were reinseminated in estrus (E-GGPG=23.7, GGPG=49.0, E-OV56=41.6, and OV56=57.6%). Treatment did not affect P/AI at 31 or 66 d for cows reinseminated in estrus. However, cows reinseminated in estrus had greater P/AI at 31 (40.0 vs. 27.5%) and 66 d (36.0 vs. 23.9%) than cows completing the TAI protocols. Among cows completing the TAI protocols, initiation of GGPG at 24 d after AI increased, whereas initiation of Ovsynch56 at 24 d after AI decreased P/AI at 31 d after reinsemination (E-GGPG=30.6, GGPG=28.3.0, E-OV56=22.3, and OV56=28.7%). Pregnancy per AI did not differ across treatment at 66 d after TAI (E-GGPG=26.6, GGPG=24.4, E-OV56=20.0, and OV56=24.1%). Overall, type of resynchronization protocol and protocol initiation time did not affect P/AI 66 d after reinsemination (E-GGPG=29.7, GGPG=30.5, E-OV56=26.1, and OV56=30.4%). In conclusion, GGPG resynchronization protocols and initiation of resynchronization protocol 24 d after AI reduced the number of cows reinseminated in estrus but neither the timing of initiation of resynchronization nor presynchronization with GnRH affected overall P/AI.

Evaluation of presynchronized resynchronization protocols for lactating dairy cows.

The objectives of this experiment were to determine the speed at which cows that had their estrous cycle presynchronized with a GnRH or PGF(2α) injection are reinseminated and become pregnant. Furthermore, this experiment aimed to determine whether treatment with a controlled internal drug-releasing (CIDR) insert during the timed artificial insemination (AI) protocol improves pregnancy per AI (P/AI) of cows that had their estrous cycle presynchronized with GnRH or PGF(2α). Lactating cows from 2 herds were assigned to 1 of 2 presynchronization treatments at 32 ± 4 d after AI: GGPG (n=452)--GnRH injection at enrollment (d 0), 7d before the start of the timed AI protocol, and P11GPG (n=466)--PGF(2α) injection on d 3, 11 d before the start of the timed AI protocol. Cows observed in estrus at any interval after enrollment were reinseminated on the same day. Cows not observed in estrus by d 7 were paired by presynchronization treatment and assigned to receive or not receive a CIDR insert during the timed AI protocol (CIDR = 240, no CIDR = 317). Timed AI protocols were the Ovsynch56 at site A and the Cosynch48 at site B. A subsample of cows from site A had their ovaries scanned by ultrasound at enrollment and on the day of the first GnRH and PGF(2α) injections of the timed AI protocol and had blood sampled at each injection of the timed AI protocol for determination of progesterone concentration. Cows were examined for pregnancy 32 ± 4 and 67 ± 4 d after reinsemination. Cows in the P11GPG treatment had a faster reinsemination rate [adjusted hazard ratio = 1.24 (95% CI = 1.07, 1.45)] and were less likely to be submitted to the timed AI protocol (40.3 vs. 89.8%) and to be reinseminated at a fixed time (38.6 vs. 83.9%). The interval from enrollment to reinsemination was shorter for cows in the P11GPG group (13.0 ± 0.4 vs. 15.0 ± 0.2d). Presynchronization treatment did not affect P/AI 32 ± 4 d (GGPG = 42.3%, P11GPG = 39.3%) and 67 ± 4 d (GGPG = 37.0%, P11GPG = 35.4%) after reinsemination. Pregnancy rate from d 0 to 7 (GGPG = 3.6%, P11GPG = 17.7%) and from d 8 to 14 (GGPG = 1.6%, P11GPG = 5.7%) were greater for cows in the P11GPG treatment. Treatment with the CIDR insert during the timed AI protocol did not affect P/AI 32 ± 4 d (CIDR = 41.7%, no CIDR = 41.4%) and 67 ± 4 d (CIDR = 36.5%, no CIDR = 35.3%) after reinsemination. A greater percentage of cows in the GGPG treatment had progesterone concentration ≥ 1 ng/mL on the day of the first GnRH injection of the timed AI protocol (83.8 vs. 51.5%), but a greater percentage of cows in the P11GPG treatment ovulated in response to the first GnRH injection of the timed AI protocol (66.1 vs. 46.8%). We conclude that the P/AI of cows that had their estrous cycle presynchronized with GnRH or PGF(2α) was not different, but in herds with adequate estrous detection efficiency and accuracy, presynchronization with PGF(2α) may reduce the interval to the establishment of pregnancy.