Pregnancy rate in water buffalo following fixed-time artificial insemination using new or used intravaginal devices with two progesterone concentrations.

This study evaluated the pregnancy rate (PR) after timed artificial insemination (TAI) in water buffalo (Bubalus bubalis) during both non-breeding and breeding season, using either a new or reused intravaginal device (IVD) with two different progesterone concentrations. A total of 247 dairy buffalo cows were randomly assigned using a two-by-three factorial design and four replicates to the following groups: (1) new intravaginal device (IVD-New: DIB®, 1.0 g of P4, n = 51 or CIDR®, 1.38 g of P4, n = 55); (2) intravaginal device previously used once (9 days) (IVD-Used1x: DIB, n = 40 or CIDR, n = 51); or (3) intravaginal device previously used twice (18 days) (IVD-Used2x: DIB, n = 27 or CIDR, n = 23). On day 0, animals received the IVD plus 10.5 µg of buserelin acetate (GnRH) intramuscularly. On day 9, the devices were removed and 25 mg of PGF2α plus 500 IU of eCG was given intramuscularly. On day 11 (48 h after IVD withdrawal), animals received 10.5 µg of GnRH and were artificially inseminated 8-12 h later. Data were analyzed using Proc Logistic of SAS®. Animals that received IVD-New-DIB, had a significantly higher PR (62.7%; P = 0.0193) compared to animals that received IVD-New-CIDR (40%). Pregnancy rate was not negatively affected by reusing both types of IVD. Overall PR (new and reused devices) was higher (P = 0.0055) in the DIB group (62.7%) compared to the CIDR group (45%). In conclusion, PR was higher in buffaloes treated with devices containing 1.0 g of P4 (DIB®) compared to those receiving 1.38 g of P4 (CIDR®). Reusing the intravaginal devices did not affect negatively PR/TAI, suggesting that P4 concentrations within the TAI protocols in water buffaloes could be reduced, without impairing their fertility.

Incidence and care-related costs of severe hypoglycaemia requiring emergency treatment in Andalusia (Spain): the PAUEPAD project.

AIMS: Hypoglycaemia is a serious medical emergency. The need for emergency medical service care and the costs of hypoglycaemic emergencies are not completely known. METHODS: This was a retrospective observational study using Public Company for Health Emergencies (EPES) data for hypoglycaemia in 2012. The EPES provides emergency medical services to the entire population of Andalusia, Spain (8.5 million people). Data on event type, onsite treatments, emergency room visits or hospitalization were collected. Medical costs were estimated using the public rates for healthcare services. RESULTS: From a total of 1 137 738 emergency calls that requested medical assistance, 8683 had a primary diagnosis of hypoglycaemia (10.34 per 10 000 person-years). The incidence of severe hypoglycaemic episodes requiring emergency treatment in the estimated population with diabetes was 80 episodes per 10 000 person-years. A total of 7479 episodes (86%) required an emergency team to visit the patient's residence. The majority of cases (64%) were addressed in the residence, although 1784 (21%) cases were transferred to hospital. A total of 5564 events (65%) involved patients aged > 65 years. Overall mortality was 0.32% (28 cases). The total annual cost of attending a hypoglycaemic episode was €6 093 507, leading to an estimated mean direct cost per episode of €702 ± 565. Episodes that required hospital treatment accounted for 49% of the total costs. CONCLUSIONS: Hypoglycaemia is a common medical emergency that is associated with high emergency medical service utilization, resulting in a significant economic impact on the health system.

Changes in serum testosterone and anti-Müllerian hormone concentration in bulls undergoing scrotal insulation.

While changes in semen quality after heat stress are well characterized in the bull, changes in endocrine function have not been critically evaluated. It was hypothesized here that scrotal insulation results in alterations in Sertoli cell and Leydig cell function, as measured by changes in serum testosterone and anti-Müllerian hormone (AMH) concentration. Scrotal insulation bags were placed in 10 bulls for 8 d. Blood was collected on days -22 and -2, and weekly from days 5 to 96 (day 0 = first day of scrotal insulation) for measurement of serum concentration of AMH and testosterone using ELISA. The concentration of AMH decreased on day 5, followed by an increase on day 54 (P = 0.014). When AMH concentration was normalized to pre-insulation values, the percent increase in serum concentration of AMH was significant between days 26 and 54, with another peak at 75 d (P = 0.031). The serum concentration of testosterone (P = 0.0001) and the percentage of change in testosterone concentration (P < 0.0001) increased on day 5, followed by a decrease from days 33 to 96. Scrotal insulation was associated with Sertoli and Leydig cell dysfunction, as measured by serum testosterone and AMH concentration. The persistently low concentration of testosterone at the end of the study suggests a long term effect of scrotal insulation on Leydig cell function.

Clinical status and endoscopy of the upper respiratory tract of dairy calves infected with Bovine viral diarrhea virus 2 and Bovine herpes virus 1 after vaccination and trace minerals injection.

The objective was to compare clinical protection [evaluated through health scoring, endoscopy score of the upper respiratory tract (URT-ES), leukocyte count, viremia, and virus shedding in nasal secretions] following Bovine viral diarrhea virus 2 (BVDV2) and Bovine herpes virus 1 (BHV1) challenge among calves submitted to modified-live virus (MLV) booster vaccination (either intranasal or subcutaneous) concurrent with injectable trace minerals (ITM) or saline. Forty-eight dairy calves received an MLV intranasal (IN) vaccine containing BHV1, BRSV, and BPI3V and subcutaneous (SC) ITM (Se, Cu, Zn & Mn; ITM, n = 24) or saline (SAL, n = 24). Ten weeks later, calves received a second dose of ITM, or saline, according to previous groups and were randomly assigned to receive the same IN vaccine [ITM-IN (n = 12), SAL-IN (n = 12)] or a SC MLV vaccine containing BHV1, BRSV, BPI3V, BVDV1 & 2 [ITM-SC (n = 12), SAL-SC (n = 12)]. Additionally, 12 calves did not receive vaccine or treatment and served as a control group (UNVAC, n = 12). Forty-nine days after booster, calves were challenged with BVDV2; and seven days later with BHV1. Health scores indicated disease in UNVAC on days 6, 10 and 12 compared to the vaccinated groups. Unvaccinated calves had the highest URT-ES after BHV1 challenge. Calves that received SC booster had lower URT-ES after BHV1 challenge than UNVAC calves. Calves in ITM-IN had significantly lower URT-ES after BHV1 infection than SAL-IN and UNVAC calves. In conclusion, IN or SC MLV vaccination was similarly effective in protecting calves from BVDV2 + BHV1 challenges (reducing clinical and endoscopy scores, preventing leukopenia, and viremia), compared to unvaccinated calves. Endoscopic evaluation of the URT allowed visualization of the inflammation and damage at multiple depths in the URT caused by a serial BVDV2 + BHV1 challenge. Calves that received SC vaccination had significantly lower URT-ES after BHV1 challenge than the UNVAC calves. Administration of ITM concurrent with IN vaccination was associated with reduced URT inflammation after BVDV2 + BHV1 challenge.

Sperm-bound antisperm antibodies are associated with poor cryosurvival of stallion spermatozoa.

Different stallions exhibit a high level of variation in the ability of their sperm to survive cryopreservation. A large fraction of stallions show poor post-thaw sperm motility, and their semen is not suitable for commercial freezing. In this study, we hypothesized that the presence of sperm-bound antisperm antibodies (ASAs) was associated with poor cryosurvival of stallion sperm. Our objective was to assess the level of ASA binding to stallion sperm, and determine if it was associated with good or poor sperm cryosurvival. In Experiment 1, cooled shipped semen from 27 stallions was frozen using three commercial semen extenders. Sperm motility, membrane integrity, acrosome integrity and apoptosis were evaluated before and after freezing for each aliquot. In addition, the percentage of ASA-bound sperm was evaluated post-thaw. In Experiment 2, semen from 22 stallions was frozen immediately after collection a single formulation of semen extender. Sperm motility and ASA binding were evaluated post-thaw. The results of both experiments showed similar findings. The frequency of ASA-positive samples was higher among stallions with poor sperm cryosurvival (Exp. 1 and 2 = 6/11, 54.5%) than for good sperm cryosurvival (Exp. 1 = 0/16, 0%; Exp. 2 = 1/11, 9.1%). The percentage of IgG- and IgA-bound sperm was also higher in stallions with poor sperm cryosurvival in both experiments (P < 0.05). Post-thaw sperm motility, velocity and distance parameters were lower in ASA-positive than ASA-negative stallions (P < 0.005). No effect of the semen extender used was observed. In addition, stallions with ASAs had a higher percentage of apoptotic sperm than stallions without ASAs. The presence of sperm-bound ASAs was associated with poor cryosurvival for stallion spermatozoa. Thus, it may be beneficial to evaluate stallions for binding of ASAs prior to freezing to offer and indicator of the prognosis for cryosurvival.

Optimization of cryopreservation protocols for cooled-transported stallion semen.

Freezing cooled-transported semen allows veterinarians and breeders to collect and process the semen of stallions on farm, and then ship the semen to a semen freezing center. There, however, is a lack of standardization of shipping and freezing protocols. The objectives were to optimize and simplify protocols to freeze cooled-shipped semen. In Experiment 1, cooled-transported semen was centrifuged at room temperature or 5 °C before freezing. Sperm variables (motility, membrane integrity, acrosome integrity, membrane fluidity) were evaluated before and after freezing. Centrifugation temperature had no effect on post-thaw semen quality. In Experiment 2, cooled-transported semen was centrifuged at room temperature and cryopreserved in three semen freezing extenders. With use of the improved modified French formula, there was less post-thaw total and progressive motility compared with use of Botucrio or the improved lactose-EDTA formula (P<0.0001). Semen cryopreserved in the improved modified French formula also had a lesser percentage of sperm with intact membranes compared with lactose-EDTA, and a greater percentage of sperm with capacitation-like changes compared with Botucrio (P<0.0001). In Experiment 3, semen diluted in each extender was frozen conventionally or placed directly in a -80 °C ultra-freezer. Freezing in the ultra-freezer resulted in a lesser post-thaw sperm motility, but not membrane and acrosome integrity and capacitation-like changes. In conclusion, centrifugation and addition of freezing extender to cooled transported semen can be performed at room temperature or 5 °C. The Botucrio and lactose-EDTA formula are recommended for conventional cryopreservation of cooled-transported stallion semen as compared with the modified French formula.

Shortening the postpartum anoestrous interval in suckled crossbred dual purpose cows using progestagen intravaginal sponges plus eCG and PGF(2alpha).

One hundred and twenty-six suckled crossbred cows (Bos taurus x Bos indicus), with body condition score >or=3 (1-5 point scale), were employed in the present study to evaluate the effectiveness of intravaginal progestin-releasing sponges (IVS) for shortening anoestrous interval. Fifty-four cows were assigned to control group. Seventy-two cows were treated with IVS impregnated with 250 mg of medroxy-acetate-progesterone (MAP) as follows: day 0, IVS plus 5 mg of 17beta-E and 50 mg of MAP i.m.; day 6, 500 IU of equine chorionic gonadotropin and 25 mg prostaglandin F(2alpha) i.m.; day 8, IVS withdrawal and day 9, 1 mg 17beta-E i.m. Cows were also grouped according to postpartum days (dpp) at treatment: MAP <70 days (n = 25); control <70 days (n = 22); MAP >70 days (n = 47); control >70 days (n = 32). From IVS removal, cows were detected in oestrus and inseminated. Cows not detected in oestrus were timed artificial insemination 72 h after sponge removal. Treatment effect on oestrous rate (ER), conception rate (CR), pregnancy rate (PR) and treatment to conception intervals (TCI) and calving to conception intervals (CCI) were evaluated. The ER, CR and PR were analysed using PROC LOGISTIC, while TCI and CCI with PROC GLM of SAS. The groups MAP <70 days and MAP >70 days showed higher (p < 0.01) ER than control <70 days and control >70 days (84.0% and 76.6% vs 31.8% and 31.3% respectively). The PR was higher (p < 0.01) in MAP <70 days vs control <70 days (64.0% vs 22.7%) and also higher (p < 0.05) in MAP >70 days vs control <70 days (40.4% vs 18.8%). The TCI and CCI were shorter (p < 0.01) in MAP <70 days vs control <70 days (36.0 and 95.8 days; 95.3 and 158.6 days respectively). In conclusion, only cows treated with IVS before 70 dpp had a CCI shorter than 100 days, consequently this treatment shortened postpartum anoestrous interval in crossbred dual purpose cattle.

Comparison between the 5-day cosynch and 7-day estradiol-based protocols for synchronization of ovulation and timed artificial insemination in suckled BOS taurus BEEF cows.

The objective was to compare pregnancy per AI and follicular dynamic in suckled Bos taurus beef cows treated with either a 7-day progesterone + estradiol-based protocol or a 5-day progesterone CoSynch protocol for timed artificial insemination (TAI) during four breeding seasons. We hypothesized that estrous cycle status, days postpartum (DPP), fat depth and plasma progesterone concentration differentially modify the effect of treatments. Every year, 9 days before initiation of each breeding season, cows were randomly assigned to one of two groups. Cows in the 7-d P + E group (n = 428) received a progesterone intravaginal device (DIB) and estradiol benzoate on Day -9. On Day -2 the device was removed, and cows received cloprostenol and estradiol cypionate. Forty-eight hours later (Day 0) cows received TAI. Cows in the 5-d P + CoS group (n = 428) received a DIB, and GnRH on Day -8. On Day -3, the device was removed, and cows received cloprostenol. A second dose of cloprostenol was given on Day -2. Cows received GnRH and TAI 72 h after device removal (Day 0). On Day -9, estrous cycle status was determined. In a subset of cows (n = 79) the size of the dominant follicle was determined between Days -2 and 0. In another subset of cows (n = 340), DPP, fat depth (mm) and plasma progesterone concentration (ng/mL) were evaluated on Day -9. Pregnancy per AI was determined 30 d after TAI. Pregnancy per AI was greater for cows in the 5-d P + CoS group than for cows in the 7-d P + E group (50.9% vs. 41.3%, P = 0.01) and was also greater in cyclic than in anestrus cows (54.3% vs. 33.2%, P < 0.0001). There was also a significant effect of breeding season (P = 0.0002) and sire (P = 0.03), and an interaction between treatment group and breeding season (P = 0.03). The dominant follicle was larger (P < 0.0001) in cows in the 5-d P + CoS group than the 7-d P + E group (10.7 ± 0.29 mm vs. 9.0 ± 0.28 mm). Pregnancy per AI was greater in cows with ≥55 DPP (47.0% vs. 29.6%, P = 0.001), fat depth ≥0.50 mm (44.7% vs. 29.7%), and with plasma progesterone concentration ≥1 ng/mL (47.2% vs. 28.7%, P = 0.01). In cows with plasma progesterone ≥1 ng/mL on Day -9, pregnancy per AI was greater in the 5-d P + CoS group (60.5%) than in the 7-d P + E group (34.9%), but there was no difference between treatment groups in cows with plasma progesterone < 1 ng/mL (P = 0.07). In conclusion, the 5-d P + CoS protocol resulted in greater size of the dominant follicle and pregnancy per AI in suckled Bos taurus beef cows subjected to TAI.

Macrophage polarization as a novel weapon in conditioning tumor microenvironment for bladder cancer: can we turn demons into gods?

Macrophages are major components of the immune infiltration in cancer where they can affect tumor behavior. In the bladder, they play important roles during the resolution of infectious processes and they have been associated with a worse clinical prognosis in bladder cancer. The present review focused on the characteristics of these important immune cells, not only eliciting an innate immune surveillance, but also on their importance during the cancer immunoediting process. We further discuss the potential of targeting macrophages for anticancer therapy, the current strategies and the state of the art as well as the foreseen role on combined therapies on the near future. This review shows how a comprehensive understanding of macrophages within the tumor should translate to better clinical outcome and new therapeutic strategies focusing especially on bladder cancer.

Control of postpartum anestrous with an intra-vaginal progesterone device plus eCG or calf removal for 120 h in suckled crossbred cows managed in a pasture-based system.

To control postpartum anestrus and reduce calving to conception interval, 167 crossbred non-pregnant cows that were 90-130 days postpartum were allotted randomly to one of the following treatments: PH (n=59), intra-vaginal sponge with 250 mg of medroxyprogesterone acetate (MAP) for 7 days plus 50mg of MAP and 5mg 17-beta estradiol (17beta-E) in the first day of treatment (day -8), 500 UI eCG (day -3) and 1.5mg 17beta-E in 24h after sponge removal (day 0); CR (n=57), temporary calf removal for 120 h; CG (n=51), control group without treatment. Estrus rate differed among treatments (P<0.01) being greater in PH (78.2%), followed by CR (52.0%) and CG (22.9%). A greater proportion of cows in the PH (80.0%) and CR (54%) groups had ovulations when compared to CG (35.4%). Intervals to first estrus were 13.5+/-6.3 days, 26.1+/-6.4 days and 52.5+/-7.5 days for the PH, CR and CG groups, respectively. First insemination conception was similar in the three groups. Postpartum intervals to first breeding (PFS) and to conception (PCI) were longer in CG than PH and CR groups (P<0.05; P<0.01). The PH and CR groups had a similar PFS but PCI was different (P<0.02). Accumulated pregnancy rate at 30 and 60 but not at 90 days were different (30 days: P<0.09; P<0.01; P<0.09; 60 days: P<0.06; P<0.01; P<0.03) among treatments. After 90 days post-treatment, 9%, 18% and 33% of cows from the PH, CR and CG groups had not conceived. Similarly, 5.4%, 6.0% and 12.5% of cows from the PH, CR and CG groups, respectively, were culled from the herd because of lack of pregnancy after 180 days post treatment. In the group of cows evaluated by ultrasonography, only those cows having larger ovaries and dominant follicles had ovulations. It was concluded that the hormonal treatment was more efficient in inducing a fertile estrus and reducing calving to conception interval followed by the calf removal for 120 h. Each method can be considered as an important tool to reduce the postpartum anestrous period in dual purpose herds when AI is conduct in the tropics.

Aerobic uterine isolates and antimicrobial susceptibility in mares with post-partum metritis.

BACKGROUND: The potential for life-threatening complications of metritis requires prompt initiation of antimicrobial treatment, often before microbiological test results are reported. However, published studies to guide first-line antibiotic selection are scarce. OBJECTIVES: To report the most frequent bacterial species, antimicrobial susceptibility and prevalence of multidrug resistance (MDR) in cases of equine metritis. STUDY DESIGN: Retrospective analysis of uterine bacterial culture and antimicrobial susceptibility test (AST) results from mares with metritis. METHODS: Data from uterine culture and AST from 45 mares and 88 bacterial isolates were analysed. The frequency of single and mixed infections, bacterial species and susceptibility to antimicrobials was reported. RESULTS: Mixed growth (62.2% mares) was more frequent than pure growth. The most commonly isolated bacterial species was Escherichia coli (30.7%) and mares with mixed growth most commonly grew a combination of Gram-negative and Gram-positive bacteria (65.5%). Gentamicin with penicillin was an appropriate choice for 65.1% of the mares. Trimethoprim/sulfonamide was effective in only 48.8% of the mares. Effective antimicrobial therapy was provided by the combination of penicillin with amikacin (90.7%) or with enrofloxacin (81.4%). Some organisms were also sensitive to tetracyclines, cephalosporins and chloramphenicol. MDR was more frequent in Gram-negative (85.4%) than Gram-positive bacteria (23.5%). MAIN LIMITATIONS: Low number of bacteria, limited number of veterinary specific interpretive criteria for equine metritis. CONCLUSIONS: Mixed infections were common in equine metritis. Gram-negative bacteria and enterococci were often associated with resistance to the most frequently used antimicrobials. Potentiated sulfonamides are not an appropriate first choice for mares with metritis. While the combination of penicillin and gentamicin may be an appropriate first-line treatment for some mares, use of amikacin or enrofloxacin instead of gentamicin may be preferential.

Effects of injectable trace minerals administered concurrently with a modified live virus vaccine on long-term protection against bovine viral diarrhea virus acute infection in dairy calves.

The objective was to evaluate the effects of injectable trace minerals (ITM) concurrent with modified-live virus (MLV) vaccination on protection from bovine viral diarrhea virus (BVDV) infection in dairy calves. In a previous study (Palomares et al., 2016), thirty dairy calves received two doses of a MLV vaccine subcutaneously (SC), concurrently with ITM (n = 15) or saline (n = 15), SC. Five months later, 20 of these calves received ITM (G1, n = 10) or saline (G2, n = 10) according to their previous groups and were challenged intranasally with BVDV2. Five unvaccinated calves were also challenged with BVDV2 (G3). Blood samples were collected on days 0 (BVDV challenge), 3, 5, 6, 7, 8, 9, 11, 14, 18, 21, 32 and 61 for leukocyte count, virus isolation and BVDV serum neutralizing antibodies (SNA). Mild-moderate clinical signs were observed in G3 after BVDV challenge. Group 1 showed lower sum health score and nasal score on d5 and fecal score on d8 compared to G2. Rectal temperature and leukocyte counts were not different between G1 and G2. In contrast, G3 calves had significant leukopenia and lymphopenia from d3 to d7 (P < .05) and higher rectal temperatures on d6 to d8, compared to values on d0 (P < .05). All unvaccinated calves became viremic, while viremia was not detected in G1 or G2. Average daily gain was not different between vaccinated groups, however, only G1 calves had significantly greater (P = .04) ADG compared to non-vaccinated calves during the first 14 days post challenge. Vaccinated calves treated or not with ITM were protected from BVDV2 infection five months post-vaccination.

Serum, uterine, and vaginal mucosal IgG antibody responses against Tritrichomonas foetus after administration of a commercial killed whole T foetus vaccine in beef cows.

The objective of this study was to determine the level and duration of IgG antibodies induced against killed whole Tritrichomonas foetus and T foetus-purified surface antigen (TF1.17) in serum, vaginal, and uterine secretions after systemic immunization of beef cows with a vaccine containing killed whole T foetus. Twenty nonpregnant beef cows were randomly assigned to vaccine or control groups as follows: Vaccine (n = 10): cows received 2 mL of a commercial vaccine containing killed whole T foetus subcutaneously and a 2-mL booster 2 weeks later. Control (n = 10): cows received 2 mL of sterile saline on the same schedule. Vaginal secretions and blood samples were collected on Days 0, 8, 15, 22, 29, 36, 43, 50, 60, 75, 89, 110, 146, and 182 relative to day of primary vaccination. Uterine flush fluid was collected on Days 0, 15, 29, and 43 after the day of primary vaccination. Samples were assayed for IgG antibodies to the killed whole T foetus and surface antigen TF1.17 using enzyme-linked immunosorbent assay. Serum whole T foetus-specific IgG levels were significantly increased (between Days 15 and 182) following vaccination with T foetus or with saline. No differences between vaccinates and controls in uterine responses to whole-cell antigen were detected. Serum anti-TF1.17 IgG responses to vaccination were significantly higher than Day 0 throughout the immunization period (P < 0.001) and were higher than responses in control animals on each day post immunization through Day 146 (P < 0.001). A significant rise in TF1.17-specific IgG levels was observed in vaginal and uterine fluids from Day 15 post vaccination compared to the Day 0 levels. These levels remained significantly elevated in vaginal and uterine fluids through Days 75 (P < 0.05) and 43 (P < 0.001) after primary vaccination, respectively. Antibody levels in serum, vaginal, and uterine secretions against TF1.17 remained low in the control group throughout the study. In conclusion, vaccination of beef cows with a commercial vaccine containing T foetus induced significant increase in the levels of IgG to the T foetus TF1.17 surface antigen in serum, vaginal secretions, and uterine fluid, which remained elevated through Days 43, 75, and 182 in uterine fluids, vaginal secretions, and serum, respectively. Since purified TF1.17 antigen has been shown to protect against experimental T foetus infection in heifers, the vaccine-induced TF1.17-specific IgG response is likely to be important in the prevention of trichomoniasis in beef cattle.

Sperm-bound antisperm antibodies prevent capacitation of bovine spermatozoa.

It was hypothesized here that sperm-bound antisperm antibodies (ASAs) impair the ability of bovine spermatozoa to undergo capacitation, bind to the zona pellucida, and complete the acrosome reaction. The effect of ASA binding on these functions was evaluated in frozen/thawed spermatozoa from four bulls before and after induction of ASAs. Ejaculates were divided into ASA negative (<10% immunoglobulin [Ig]G- and IgA-bound spermatozoa) or ASA positive (≥20% IgG and/or IgA-bound spermatozoa). The percentage of capacitated (Merocyanine 540 positive) live spermatozoa in response to heparin was lower in ASA-positive than ASA-negative ejaculates (P < 0.0001). Treatment with heparin resulted in a higher percentage of capacitated spermatozoa compared with control treatments in ASA-negative but not ASA-positive ejaculates. The percentage of capacitated spermatozoa after heparin treatment was negatively correlated with IgA (P = 0.02, R2 = -0.48) but not IgG binding. Sperm binding to the zona pellucida was lower in IgA-positive (six spermatozoa/oocyte; 3-10 spermatozoa/oocyte) than IgA-negative ejaculates (seven spermatozoa/oocyte; 4-13 spermatozoa/oocyte) (P = 0.019). Zona binding was negatively correlated with the percentage of IgA-bound spermatozoa (P = 0.04; R2 = -0.24) but not IgG-bound spermatozoa. The percentage of acrosome-reacted spermatozoa was higher in calcium ionophore A23187-treated than control aliquots in both ASA-negative and ASA-positive ejaculates (P < 0.0001). However, the percentage of acrosome-reacted spermatozoa did not differ between ASA-positive and ASA-negative samples, and no correlation was identified with IgG or IgA binding. It was concluded that sperm-bound IgA affected the ability of bovine spermatozoa to undergo capacitation. ASAs inhibited the changes in plasma membrane fluidity associated with capacitation and binding of spermatozoa to the zona pellucida.

Effects of injectable trace minerals on humoral and cell-mediated immune responses to Bovine viral diarrhea virus, Bovine herpes virus 1 and Bovine respiratory syncytial virus following administration of a modified-live virus vaccine in dairy calves.

Our objective was to evaluate the effect of an injectable trace mineral (ITM) supplement containing zinc, manganese, selenium, and copper on the humoral and cell mediated immune (CMI) responses to vaccine antigens in dairy calves receiving a modified-live viral (MLV) vaccine containing BVDV, BHV1, PI3V and BRSV. A total of 30 dairy calves (3.5 months of age) were administered a priming dose of the MLV vaccine containing BHV1, BVDV1 & 2, BRSV, PI3V, and an attenuated-live Mannheimia-Pasteurella bacterin subcutaneously (SQ). Calves were randomly assigned to 1 of 2 groups: (1) administration of ITM SQ (ITM, n=15) or (2) injection of sterile saline SQ (Control; n=15). Three weeks later, calves received a booster of the same vaccine combination SQ, and a second administration of ITM, or sterile saline, according to the treatment group. Blood samples were collected on days 0, 7, 14, 21, 28, 42, 56, and 90 post-vaccination for determination of antibody titer, viral recall antigen-induced IFN-γ production, and viral antigen-induced proliferation by peripheral blood mononuclear cells (PBMC). Administration of ITM concurrently with MLV vaccination resulted in higher antibody titers to BVDV1 on day 28 after priming vaccination compared to the control group (P=0.03). Calves treated with ITM showed an earlier enhancement in PBMC proliferation to BVDV1 following vaccination compared to the control group. Proliferation of PBMC after BVDV stimulation tended to be higher on day 14 after priming vaccination in calves treated with ITM than in the control group (P=0.08). Calves that received ITM showed higher PBMC proliferation to BRSV stimulation on day 7 after priming vaccination compared to the control group (P=0.01). Moreover, calves in the ITM group also had an enhanced production IFN-γ by PBMC after stimulation with BRSV on day 21 after priming vaccination compared to day 0 (P<0.01). In conclusion, administration of ITM concurrently with MLV vaccination in dairy calves resulted in increased antibody titer to BVDV1, and greater PBMC proliferation to BVDV1 and BRSV recall stimulation compared to the control group, suggesting that ITM might represent a promising tool to enhance the humoral and CMI responses to MLV vaccines in cattle.

Mechanism of endogenous phosphorylation of microtubule proteins during GTP-induced microtubule assembly and implications for stability of the assembled structures.

Cycle-purified microtubule protein from mammalian brain incorporated [32P]Pi upon incubation with [gamma-32P]GTP under the conditions used to promote assembly. This phosphorylation also occurred in the same proteins when phosphorylated with [gamma-32P]ATP and was only slightly stimulated by cAMP. GTP was a much less effective substrate than ATP. The transfer of phosphoryl groups from [gamma-32P]GTP to endogenous proteins followed a linear time-course and was stimulated by low concentrations of ATP and, more efficiently, by ADP. These data are in agreement with the predictions derived from a mechanism of phosphorylation by which [gamma-32P]GTP does not act as a phosphoryl donor for the protein kinase activity but, instead, only as a repository of high group transfer potential phosphoryl groups used to make [gamma-32P]ATP, from contaminating ADP, by means of the nucleoside diphosphate kinase activity. Using 100 mM fluoride, which suppressed protein phosphorylation without inhibiting the nucleoside diphosphate kinase activity, formation of [gamma-32P]ATP was detected. Fluoride was also able to protect microtubules from a slow depolymerization which was found to occur during long-term incubation of microtubules. This indicates that the phosphorylation observed in the presence of GTP is sufficient to destabilize microtubules.

Measuring the noble metal and iodine composition of extracted noble metal phase from spent nuclear fuel using instrumental neutron activation analysis.

Masses of noble metal and iodine nuclides in the metallic noble metal phase extracted from spent fuel are measured using instrumental neutron activation analysis. Nuclide presence is predicted using fission yield analysis, and radionuclides are identified and the masses quantified using neutron activation analysis. The nuclide compositions of noble metal phase derived from two dissolution methods, UO2 fuel dissolved in nitric acid and UO2 fuel dissolved in ammonium-carbonate and hydrogen-peroxide solution, are compared.

Different blue-light requirement for the accumulation of transcripts from nuclear genes for thylakoid proteins in Nicotiana tabacum and Lycopersicon esculentum.

We have isolated recombinant lambda gt11 phages which carry cDNA clones for the major light-harvesting chlorophyll a/b-binding proteins of photosystem I (LHCPI) and II (LHCPII), subunit II of photosystem I, a chlorophyll a/b-binding protein of photosystem II (CP24), the Rieske iron-sulphur protein of the cytochrome b6/f complex, and the 33, 23 and 16 kDa proteins of the water-oxidizing complex of photosystem II from Nicotiana tabacum. The nucleotide sequences of cDNA clones encoding the precursors for LHCPI and the FeS protein are presented. If tobacco or tomato seedlings, or seedlings of a phytochrome-deficient aurea mutant of tomato which lacks more than 95% of the phytochrome of the isogenic wild type, are kept in blue light, the transcript level of each of these genes is higher than in seedlings grown in red light suggesting the involvement of a blue-UVA-light photoreceptor. In the case of LHCPI, a 1 min blue-light pulse applied to red-light-grown seedlings is sufficient to increase the transcript levels to those present in blue-light-grown seedlings, whereas almost no increase is observed for transcripts encoding the FeS and 33 kDa proteins. If dark-grown tomato seedlings receive a single far-red-light pulse, significant stimulation is detected for LHCPI transcripts, whereas transcripts encoding the FeS and 33 kDa proteins are not stimulated. It is concluded that the lower light requirement for the increase in the LHCPI transcript level is not specific for one of the light-dependent signal transduction chains.

The effects of intrauterine cocaine exposure: transient or teratogenic?

The use of cocaine and its smoked derivative, crack, is increasing in America. Among those affected are neonates of women who use cocaine during pregnancy. This paper examines the potential effects of maternal cocaine use upon the developing fetus emphasizing neuropsychological and behavioral effects including those that typically persist into early childhood and beyond. Empirical research should continue to explore the effects of maternal cocaine use focusing on whether or not the many detrimental effects of cocaine use during pregnancy are transient, directly teratogenic, or indirect and due to prolonged hypoxia.