Retained energy in lactating beef cows; effects on maintenance energy requirement and voluntary feed intake.

The objectives of these experiments were to determine the relationship between maintenance requirements and energy partitioned to maternal tissue or milk production in limit-fed Angus cows and to determine the relationship between retained energy during the lactation period to dry-period voluntary forage intake (VDMI). Twenty-four mature fall-calving Angus cows were used in a 79-d study during late lactation to establish daily metabolizable energy required for maintenance (MEm). Cows were individually fed daily a mixed diet (2.62 Mcal MEl/kg, 18.2% crude protein) to meet energy and protein requirements of 505 kg beef cows producing 8.2 kg milk daily. If cow BW changed by ±9 kg from initial BW, daily feed intake was adjusted to slow BW loss or reduce BW gain. Milk yield and composition were determined on 3 occasions throughout the study. Maintenance was computed as metabolizable energy intake minus retained energy assigned to average daily maternal tissue energy change, average daily milk energy yield, and average daily energy required for pregnancy. After calves were weaned, cows were fed a low-quality grass hay diet (8.2% crude protein, 65% NDF) and VDMI was measured for 21 days. Lactation maintenance energy was 83% the default value recommended by NASEM (2016. Nutrient Requirements of Beef Cattle: Eighth Revised Edition.) for lactating Angus cows. Increasing lactation-period retained energy (decreasing BW loss and increasing milk energy yield) was associated with lower maintenance energy requirements (P < 0.01; R 2 = 0.92). Increased residual daily gain during lactation was associated with lower lactation maintenance energy requirements (P = 0.05; R 2 = 0.17). Post-weaning VDMI was not related to late-lactation milk energy production, although sensitive to lactation period BCS and BW loss. These results contradict previous reports, suggesting that maintenance requirements increase with increasing milk yield.

Effect of vitamin D source and dietary cation-anion difference in peripartum dairy cows on calcium homeostasis and milk production.

The objective of this experiment was to determine the effects of dietary vitamin D source on serum calcium (Ca), urinary Ca excretion, and milk production when fed in combination with a prepartum acidogenic negative dietary cation-anion difference (DCAD) diet. Nonlactating, pregnant multiparous cows (n = 15), balanced for breed (Holstein n = 9 and Jersey n = 6), and previous mature equivalent milk production, were assigned to one of three treatments (five cows/treatment), consisting of a control (PCH; positive DCAD, 8.9 mEq/100 g DM) and two negative DCAD diets (-15.4 mEq/100 g DM), one with vitamin D3 (cholecalciferol; NCH) and one with 25-hydroxyvitamin D3 (calcidiol; NCA; DSM nutritional products). The treatments were formulated to provide 1.95 mg/d of vitamin D and were fed 28 d prior to expected calving date. Delivery of vitamin D sources was accomplished by manufacture of a pellet and 2 kg of these pellets were individually fed simultaneously each day along with 2 kg of ground corn daily at 0800 hours. Negative DCAD treatments were formulated to provide 0.46 kg/d of Animate (Phibro Animal Health) and, if needed, additional Animate was top-dressed at each feeding to achieve a urine pH between 5.5 and 6.0 based on the previous day's urine pH. Close-up cows had ad libitum access to chopped bermudagrass (Cynodon dactylon L.) hay and hay intake was measured using SmartFeed Pro systems (C-Lock Inc.; Rapid City, SD). Prepartum urine and serum samples were collected weekly and serum was collected 36, 48, and 72 h post-calving. Prepartum dry matter intake (DMI) as a percent of body weight was not (P = 0.66) affected by treatments. Cows fed NCH and NCA had greater (P = 0.02) prepartum serum Ca than PCH and tended to have greater urinary Ca excretions (P = 0.10). Average postpartum serum Ca (mg/dL) was greater (P = 0.05) for cows fed NCH (8.8) compared with PCH (7.8), whereas NCA (8.4) was numerically intermediate and not (P > 0.05) different from either of the other treatments. Postpartum DMI was not affected by treatment (P = 0.39). Daily milk yield (MY) (kg/d) was greatest (P < 0.01) for NCA (37.5) compared with the other treatments and NCH (34.1) was intermediate and greater than PCH (29.9). These results suggest that an acidogenic prepartum diet in combination with vitamin D was effective in maintaining peripartum serum Ca and the 25-hydroxy form of vitamin D improved MY compared with NCH in early lactation.

Technical note: using an automated head chamber system to administer an external marker to estimate fecal output by grazing beef cattle.

The objective of this experiment was to determine if titanium dioxide (TiO2) dosed through an automated head chamber system (GreenFeed; C-Lock Inc., Rapid City, SD, USA) is an acceptable method to measure fecal output. The GreenFeed used on this experiment had a 2-hopper bait dispensing system, where hopper 1 contained alfalfa pellets marked with 1% titanium dioxide (TiO2) and hopper 2 contained unmarked alfalfa pellets. Eleven heifers (BW = 394 ± 18.7 kg) grazing a common pasture were stratified by BW and then randomized to either 1) dosed with TiO2-marked pellets by hand feeding (HFD; n = 6) or 2) dosed with TiO2-marked pellets by the GreenFeed (GFFD; n = 5) for 19 d. During the morning (0800), all heifers were offered a pelleted, high-CP supplement at 0.25% of BW in individual feeding stanchions. The HFD heifers also received 32 g of TiO2-marked pellets at morning feeding, whereas the GFFD heifers received 32 g of unmarked pellets. The GFFD heifers received a single aliquot (32 ± 1.6 g; mean ± SD) of marked pellets at their first visit to the GreenFeed each day with all subsequent 32-g aliquots providing unmarked pellets; HFD heifers received only unmarked pellets. Starting on d 15, fecal samples were collected via rectal grab at feeding and every 12 h for 5 d. A two-one sided t-test method was used to determine agreement and it was determined that the fecal output estimates by HFD and GFFD methods were similar (P = 0.04). There was a difference (P < 0.01; Bartlett's test for homogenous variances) in variability between the dosing methods for HFD and GFFD (SD = 0.1 and 0.7, respectively). This difference in fecal output variability may have been due to variability of dosing times-of-day for the GFFD heifers (0615 ± 6.2 h) relative to the constant dosing time-of-day for HFD and constant 0800 and 2000 sampling times-of-day for all animals. This research has highlighted the potential for dosing cattle with an external marker through a GreenFeed configured with two (or more) feed hoppers because estimated fecal output means were similar; however, consideration of the increased variability of the fecal output estimates is needed for future experimental designs.

Microwave-Assisted Cross-Polarization of Nuclear Spin Ensembles from Optically Pumped Nitrogen-Vacancy Centers in Diamond.

The ability to optically initialize the electronic spin of the nitrogen-vacancy (NV) center in diamond has long been considered a valuable resource to enhance the polarization of neighboring nuclei, but efficient polarization transfer to spin species outside the diamond crystal has proven challenging. Here we demonstrate variable-magnetic-field, microwave-enabled cross-polarization from the NV electronic spin to protons in a model viscous fluid in contact with the diamond surface. Further, slight changes in the cross-relaxation rate as a function of the wait time between successive repetitions of the transfer protocol suggest slower molecular dynamics near the diamond surface compared to that in bulk. This observation is consistent with present models of the microscopic structure of a fluid and can be exploited to estimate the diffusion coefficient near a solid-liquid interface, of importance in colloid science.

Coherent properties of single rare-earth spin qubits.

Rare-earth-doped crystals are excellent hardware for quantum storage of photons. Additional functionality of these materials is added by their waveguiding properties allowing for on-chip photonic networks. However, detection and coherent properties of rare-earth single-spin qubits have not been demonstrated so far. Here we present experimental results on high-fidelity optical initialization, efficient coherent manipulation and optical readout of a single-electron spin of Ce(3+) ion in a yttrium aluminium garnet crystal. Under dynamic decoupling, spin coherence lifetime reaches T2 = 2 ms and is almost limited by the measured spin-lattice relaxation time T1 = 4.5 ms. Strong hyperfine coupling to aluminium nuclear spins suggests that cerium electron spins can be exploited as an interface between photons and long-lived nuclear spin memory. Combined with high brightness of Ce(3+) emission and a possibility of creating photonic circuits out of the host material, this makes cerium spins an interesting option for integrated quantum photonics.

High-precision nanoscale temperature sensing using single defects in diamond.

Measuring local temperature with a spatial resolution on the order of a few nanometers has a wide range of applications in the semiconductor industry and in material and life sciences. For example, probing temperature on the nanoscale with high precision can potentially be used to detect small, local temperature changes like those caused by chemical reactions or biochemical processes. However, precise nanoscale temperature measurements have not been realized so far owing to the lack of adequate probes. Here we experimentally demonstrate a novel nanoscale temperature sensing technique based on optically detected electron spin resonance in single atomic defects in diamonds. These diamond sensor sizes range from a micrometer down to a few tens of nanometers. We achieve a temperature noise floor of 5 mK/Hz(1/2) for single defects in bulk sensors. Using doped nanodiamonds as sensors the temperature noise floor is 130 mK/Hz(1/2) and accuracies down to 1 mK for nanocrystal sizes and therefore length scales of a few tens of nanometers. This combination of precision and position resolution, combined with the outstanding sensor photostability, should allow the measurement of the heat produced by chemical interactions involving a few or single molecules even in heterogeneous environments like cells.

Southern Section Interdisciplinary Beef Cattle Symposium: Carryover effects of stocker cattle systems on feedlot performance and carcass characteristics.

Stocker cattle production practices may affect subsequent performance of cattle in economically meaningful ways. Literature was reviewed to summarize possible carryover effects of stocker cattle management on finishing performance and carcass traits. The most consistent effects of stocker phase practices on finishing-phase and carcass performance of cattle appear to be explained by changes in placement weight of feeder cattle. Increasing placement weight typically is confounded with age at placement, ADG during stocker period, and duration of the stocker period. It may also be confounded with other factors, such as season of grazing and/or finishing. Data from studies comparing calf-fed to yearling-fed cattle indicate that as age or BW at feedlot entry increased, finishing ADG, DMI, and HCW increased (P<0.01) whereas G:F and days on feed decreased (P<0.01). As age and BW at feedlot entry increased, marbling score decreased (P<0.01) whereas Warner-Bratzler shear force increased (P<0.01). Studies with yearling cattle entering the feedyard after a forage-based backgrounding phase indicate that as backgrounding ADG increased, finishing ADG, DMI, and days on feed decreased (P<0.01) yet HCW and ribeye area increased (P<0.01) with no effect (P=0.45) on marbling. Stocker cattle systems are complex, integrated systems in which producers typically seek to maximize their own economic return with little regard for subsequent performance. If cattle are owned across both stocker and finishing phases of production, then management decisions should focus on maximizing return over the entire ownership period and not just within one segment of ownership. Of the traits of feeder cattle that can be affected by stocker cattle systems, BW is the primary determinate of their value when they are placed into commercial feedyards.

Optical detection of a single rare-earth ion in a crystal.

Rare-earth-doped laser materials show strong prospects for quantum information storage and processing, as well as for biological imaging, due to their high-Q 4f↔4f optical transitions. However, the inability to optically detect single rare-earth dopants has prevented these materials from reaching their full potential. Here we detect a single photostable Pr(3+) ion in yttrium aluminium garnet nanocrystals with high contrast photon antibunching by using optical upconversion of the excited state population of the 4f↔4f optical transition into ultraviolet fluorescence. We also demonstrate on-demand creation of Pr(3+) ions in a bulk yttrium aluminium garnet crystal by patterned ion implantation. Finally, we show generation of local nanophotonic structures and cell death due to photochemical effects caused by upconverted ultraviolet fluorescence of praseodymium-doped yttrium aluminium garnet in the surrounding environment. Our study demonstrates versatile use of rare-earth atomic-size ultraviolet emitters for nanoengineering and biotechnological applications.

Differential acute phase immune responses by Angus and Romosinuano steers following an endotoxin challenge.

Our primary objective of this experiment was to evaluate potential genetic differences between two diverse Bos taurus breeds [Angus (AG) and Romosinuano (RO)] in response to an endotoxin challenge. Eighteen steers (n = 9 steers/breed; 299.4 ± 5.2 kg BW) were acclimated to environmentally controlled chambers maintained at thermoneutrality (19.7 °C) and then fitted with indwelling jugular catheters and rectal temperature (RT) recording devices 1 d before the endotoxin challenge. The next day, blood samples were collected at 30-min intervals from -2 to 8 h, and RT was measured continuously at 1-min intervals throughout the study. At time 0, all steers received an intravenous bolus injection of lipopolysaccharide (LPS; 2.5 µg/kg BW). Serum samples were stored at -80 °C until analyzed for cortisol, proinflammatory cytokines [tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1ß), IL-6, and interferon gamma (IFN-γ)], and acute phase proteins (serum amyloid A, acid soluble protein, ceruloplasmin, and α-acid glycoprotein). Rectal temperatures increased in both breeds within 1 h after LPS, with RO producing a greater increase in RT than AG steers (P < 0.001). Serum cortisol and TNF-α increased (P < 0.01) in both breeds within 1 h after the LPS challenge. For cortisol, an overall breed effect (P < 0.02) was detected, such that AG steers had a higher cortisol response than RO steers. A breed × time interaction (P < 0.01) was observed for TNF-α, such that the response was delayed and extended in the RO steers compared with the AG steers. At 2 and 2.5 h after LPS, TNF-α concentrations were greater (P < 0.03) in RO steers than in AG steers. For IL-1ß, a breed × time interaction (P < 0.04) was also observed. At 3 h after LPS, IL-1ß concentrations were greater (P < 0.01) in RO steers than in AG steers. Serum IL-6 and IFN-γ increased (P < 0.01) in a similar manner in both groups after the LPS challenge. These data show differences in the innate immune response between two diverse Bos taurus breeds which may provide insight about differences observed in productivity, heat tolerance, disease resistance, and longevity among cattle breeds.

Relationships between fed cattle traits and Igenity panel scores.

Although several previous studies have identified associations between cattle carcass characteristics and various SNP, comparatively little work has sought to validate the marker panels currently sold and marketed by commercial genotyping companies. Panels typically use a handful of SNP, but these range from as few as 2 to more than 100. Data from 764 commercially fed steers and heifers were used to assess the relationships of growth and carcass traits and Igenity panel scores for ADG, marbling (or percentage of USDA Choice), rib-eye area (REA), tenderness, fat thickness, and USDA Yield grade (YG). Results revealed statistically significant, but low, correlations between carcass measurements and corresponding Igenity panel scores. Genetic correlations were computed among the various Igenity panels and demonstrated either that several common markers existed across the panels or that markers across panels were in high linkage disequilibrium. Across all breeds, the genotypic correlations between the Igenity panel scores for ADG, REA, marbling, and YG with observed ADG, REA, USDA Quality grade, and YG at slaughter were 0.51, 0.38, 0.63, and 0.59 (P < 0.01), respectively. The partial effects of the Igenity marbling panel persisted in a multivariate regression model. Net return was significantly affected only by marbling panel score; a 1-unit increase in marbling panel score increased the net return by an estimated $7.53 per animal.

Technical note: Development of a self-contained, indwelling rectal temperature probe for cattle research.

A device was developed to monitor rectal temperature automatically in cattle for application in research settings. Compared with manual measurement of rectal temperature, this device decreases labor and time requirements and allows data collection without the influence of animal handling or restraint. The device consists of a custom-fabricated aluminum tail harness that supports an indwelling rectal temperature data logger. Materials cost approximately US $300 per unit, and units are completely reusable. Use of this device would increase the conditions under which accurate rectal temperature measurements can be obtained in experiments with cattle.

Effects of duration of zilpaterol hydrochloride feeding and days on the finishing diet on feedlot cattle performance and carcass traits.

British and British x Continental steers (n = 560; initial BW = 339.4 +/- 1.76 kg) were used in a serial slaughter study with a completely random design to evaluate effects of zilpaterol hydrochloride (ZH; 8.33 mg/kg of dietary DM basis) on performance and carcass characteristics. Treatments were arranged in a 4 x 4 factorial (112 pens; 7 pens/treatment; 5 steers/pen) and included duration of ZH feeding (0, 20, 30, or 40 d before slaughter plus a 3-d ZH withdrawal period) and days on feed (DOF) before slaughter (136, 157, 177, and 198 d). No duration of ZH feeding x slaughter group interactions were detected for the performance measurements (P > 0.10). Final BW did not differ (P = 0.15) between the 0-d group and the average of the 3 ZH groups, but ADG was greater for the average of the 3 ZH groups during the period in which ZH diets were fed (P < 0.01) and for the overall feeding period (P = 0.05). As duration of ZH feeding increased, DMI decreased (P = 0.01) and G:F increased linearly (P < 0.01). With the exception of KPH (P = 0.022), no duration of ZH feeding x slaughter group interactions (P > 0.10) were detected for carcass characteristics. Regardless of the duration of ZH feeding, cattle fed ZH had greater HCW (P < 0.01), greater dressing percent (P < 0.01), less 12th-rib fat (P < 0.01), larger LM area (P < 0.01), less KPH (P = 0.03), and lower yield grade (P < 0.01) than the 0-d cattle. The 0-d group had greater marbling scores (P < 0.01) than cattle fed ZH diets, with a tendency for a linear decrease in marbling score (P = 0.10) as duration of ZH feeding was extended. A greater percentage of carcasses in the 0-d group graded USDA Choice or greater (P < 0.01) than in the 3 ZH groups, whereas the percentage of Select carcasses was greater (P = 0.01) for the 3 ZH groups. From d 0 to end (P = 0.04) and during the last 43 d on feed (P < 0.01), ADG responded quadratically to DOF before slaughter. No differences were detected among slaughter groups for DMI for the entire trial period; however, a quadratic response (P = 0.02) was observed for the final 43 d before slaughter. A quadratic response was also detected for the final 43 d before slaughter (P < 0.01) and from d 0 to end (P = 0.02) for G:F. Final BW, HCW, dressing percent, and 12th-rib fat increased linearly (P < 0.01) as DOF before slaughter increased. Our results indicate that no substantial effects on performance and carcass measurements were observed when ZH was fed for 30 or 40 d as opposed to 20 d, and that effects of ZH generally did not interact with DOF before slaughter.

Effects of dietary energy source and level and injection of tilmicosin phosphate on immune function in lipopolysaccharide-challenged beef steers.

Twenty-four Angus x Hereford crossbred steers (247 kg BW; SE = 2.4 kg) were used in a completely random design to evaluate the effect of energy source and level with or without antibiotic administration on measures of immune function. Steers were fed 1 of 3 dietary treatments: a 70% concentrate diet ad libitum (70AL), a 30% concentrate diet ad libitum (30AL), and a 70% concentrate diet offered in an amount calculated to provide NE(g) intake equal to the 30AL treatment (70RES). Half the steers in each dietary treatment received a s.c. injection of tilmicosin phosphate (ANTI; 1 mL/30 kg of BW); the other half received an equal volume of saline s.c. (SAL). Steers were offered the treatment diets for 28 d before and were administered the ANTI or SAL injections 2 d before indwelling catheters were placed in the jugular vein and 2.0 microg/kg of BW of Escherichia coli lipopolysaccharide (LPS) was administered i.v. Blood serum was collected at 30-min intervals from -2 to 6 h and at 8, 12, 24, 48, and 72 h relative to the LPS challenge. Increased energy intake (70AL) increased (P < or = 0.04) DMI, ADG, and rectal temperature (RT) after the challenge compared with the 70RES treatment. The 30AL treatment increased the maximum concentrations and area under the response curve of the proinflammatory cytokines (PIC) interferon-gamma, tumor necrosis factor-alpha, and IL-6 (P < or = 0.05) compared with the average of the 70AL and 70RES treatments. Decreased energy intake (70RES vs. 70AL) increased IL-6 (P < or = 0.003) but did not significantly increase interferon-gamma and tumor necrosis factor-alpha (P > or = 0.14) after LPS administration. Tilmicosin administration decreased the time to attain maximal RT (P = 0.01) by 1 h without altering the peak RT (P = 0.85), and tilmicosin interacted with energy intake to increase prechallenge PIC in 70RES vs. 70AL (P < or = 0.05). Results indicate that increased PIC response, presumably resulting from a combination of decreased energy intake and from direct effects of roughage, may be a mode of action for the slight decrease in morbidity that often occurs when newly received, stressed calves are fed roughage-based receiving diets. Tilmicosin phosphate might have immunomodulatory capacity beyond its direct effects on pathogenic bacteria, and these effects could interact with dietary energy intake in cattle.

Pancreatic mixed acinar-endocrine carcinoma in a dog.

A 10.5-year-old crossbreed dog was presented with a history of hypoglycaemic episodes and elevated serum insulin concentration. A pancreatic mass was removed at surgery along with an enlarged draining lymph node. An unresectable hepatic nodule was also present. Immunohistochemistry confirmed the pancreatic and lymph node masses as functional mixed acinar-endocrine carcinoma, previously unreported in domestic species. Persistent hypoglycaemia and hyperinsulinaemia post-operatively was highly suggestive of the hepatic mass being a functional metastasis. The dog was managed on prednisolone and remained asymptomatic 9 months post-operatively. This tumour type has only been rarely reported in human patients and may highlight the need for more rigorous immunohistochemical staining of pancreatic masses in veterinary species to identify the prevalence of this tumour type.

ParSeq: searching motifs with structural and biochemical properties.

SUMMARY: Searches for variable motifs such as protein-binding sites or promoter regions are more complex than the search for casual motifs. For example, in amino acid sequences comparing motifs alone mostly proves to be insufficient to detect regions that represent proteins with a special function, because the function depends on biochemical properties of individual amino acids (such as polarity or hydrophobicity). Pure string matching programs are not able to find these motifs; hence, we developed ParSeq, a program that combines the search for motifs with certain structural properties, the verification of biochemical properties, an approximate search mechanism and a stepwise creation of the motif description by allowing to search on previously obtained results. AVAILABILITY: http://www-pr.informatik.uni-tuebingen.de/parseq

Improved procedure of colonoscopy under accompanying music therapy.

BACKGROUND: Administration of sedatives and analgetics during colonoscopy includes the risk for arterial hypotension and respiratory depression. The aim of this study was to assess whether music therapy increases patients tolerance and reduces the need of analgo-sedative premedication. METHODS: 146 consecutive patients were examined in a randomized, prospective study. Colonoscopy was performed under intravenous administration with titrated dosages of midazolam and pethidin. Oxygen was given in cases of blood desaturation below values of 90%. Patients younger than 18 and older than 80 years, patients with history of partial colectomy, gastrectomy or hysterectomy and patients with colonic tumorous or inflammatory stenosis were excluded. 60 patients underwent conventional procedure (Group A), whereas 59 patients received additional music therapy (Group B). Time required to reach the cecum (examination time) was measured and the rate of successful colonoscopies was determined. RESULTS: Most of the patients required sedation with midazolam in both groups (97 vs. 93%), whereas more group A patients required analgesia with pethidin than group B patients (43 vs. 31%, p<0.05). Under music therapy the rate of completed colonoscopies was higher (group A 93%, group B 98%) and examination time was significantly accelerated (group A 22.8 +/- 14.6 min, group B 16.8 +/- 11.8 min, p<0.03). CONCLUSIONS: Accompanying music therapy reduces requirement of analgesia during colonoscopy, favours completion of the procedure and shortens examination time. Music therapy seems to promote safer conditions for endoscopical practice and diminishs patients discomfort.

Regulation of phosphotransferase activity of hexokinase 2 from Saccharomyces cerevisiae by modification at serine-14.

Isoenzyme 2 of hexokinase functions in sugar sensing and glucose repression in Saccharomyces cerevisiae. The degree of in vivo phosphorylation of hexokinase 2 at serine-14 is inversely related to the extracellular glucose concentration [Vojtek, A. B., and Fraenkel, D. G. (1990) Eur. J. Biochem. 190, 371-375]; however, a physiological role of the modification causing the dissociation of the dimeric enzyme in vitro [as effected by a serine-glutamate exchange at position 14; Behlke et al. (1998) Biochemistry 37, 11989-11995] is unclear. This paper describes a comparative stopped-flow kinetic and sedimentation equilibrium analysis performed with native unphosphorylated hexokinase 2 and a permanently pseudophosphorylated glutamate-14 mutant enzyme to determine the functional consequences of phosphorylation-induced enzyme dissociation. The use of a dye-linked hexokinase assay monitoring proton generation allowed the investigation of the kinetics of glucose phosphorylation over a wide range of enzyme concentrations. The kinetic data indicated that monomeric hexokinase represents the high-affinity form of isoenzyme 2 for both glycolytic substrates. Inhibition of glucose phosphorylation by ATP [Moreno et al. (1986) Eur. J. Biochem. 161, 565-569] was only observed at a low enzyme concentration, whereas no inhibition was detected at the high concentration of hexokinase 2 presumed to occur in the cell. Pseudophosphorylation by glutamate substitution for serine-14 increased substrate affinity at high enzyme concentration and stimulated the autophosphorylation of isoenzyme 2. The possible role of hexokinase 2 in vivo phosphorylation at serine-14 in glucose signaling is discussed.

Performance of light vs heavy steers grazing Plains Old World bluestem at three stocking rates.

Live weight gains of light and heavy calves grazing Plains Old World bluestem at three stocking rates were evaluated during the summers of 1997 and 1998. Initial weights of mixed-breed light-weight steers (LHT) were 141 SD = 17 kg (n = 214) in 1997 and 160 SD = 23 kg (n = 193) in 1998. Initial weights of mixed-breed heavy steers (HWT) were 265 SD = 17 kg (n = 115) in 1997 and 248 SD = 13 kg (n = 126) in 1998. Initial stocking rates for both sizes of steers were as follows: light, 392 kg of live weight/ha; moderate, 504 kg of live weight/ha (increased to 616 kg live weight/ha in 1998); and heavy, 840 kg of live weight/ ha. Averaged gain and gain/hectare are reported as stocking rate by steer type within year. Heavy steers had greater ADG than LHT steers during both years. Forage intake, expressed as a percentage of BW, was greater (P = 0.05) for LHT (3.1%) than for HWT (2.8%) calves. Grazing time (min/d; 1998 only) was greater (P = 0.05) for LHT (665) than for HWT (624) steers. Forage CP and in vivo digestible organic matter (DOM) were slightly greater (P < 0.05) in pastures grazed by HWT vs LHT cattle. Gain/hectare was greater (P < 0.05) for LHT than for HWT calves at all three stocking rates during both years. A linear decline in ADG was observed (P < 0.07) as stocking rates increased for HWT steers in 1997 and LHT steers in 1998. However, ADG did not decline with increasing stocking rate for LHT calves during 1997 or HWT calves during 1998. Forage intake was not different among stocking rates in either 1997 or 1998. Grazing time was greatest (P < 0.05) for steers in the moderate and heavy stocking rates. Forage in vivo DOM decreased (P < 0.05) as stocking rate increased. Both LHT and HWT steers had lower (P < 0.05) ADG at all three stocking rates during 1998 compared with 1997. Despite lower ADG, LHT steers had greater gain/hectare than HWT steers during both 1997 and 1998.

Purification, molecular and kinetic characterization of phosphofructokinase-1 from the yeast Schizosaccharomyces pombe: evidence for an unusual subunit composition.

Phosphofructokinase-1 (Pfk-1) from Schizosaccharomyces pombe was purified by 54-fold enrichment to homogeneity elaborating the following steps: (a) Disruption of the cells with glass beads; (b) fractionated precipitation with polyethylene glycol 6000; (c) affinity chromatography on Cibacron-Blue F3G-A-Sephadex G 100; (d) ion exchange chromatography on Resource Q. The native enzyme exhibits a mass of 790+/-30 kDa, as detected by sedimentation equilibrium measurements. The apparent sedimentation coefficient was found to be s(20,c)=20.2+/-0.3 S. No significant dependence of the s-value on the protein concentration was observed in the range 0. 07-0.7 mg/ml. Polyacrylamide gel electrophoresis in presence of sodium dodecyl sulphate and MALDI-TOF spectra showed that the enzyme is composed of subunits of identical size of 100+/-5 kDa, forming an octameric structure. The N-terminus of the enzyme was found to be blocked. Sequences of tryptic and chymotryptic peptides of the subunit coincide with the proposed amino acid sequence as deduced from the gene from the EMBL library. The Pfk-1 coding sequence of S. pombe was transformed into a Pfk-1 double deletion mutants of Saccharomyces cerevisiae resulting in glucose-positive cells with enzyme activity in the crude cell extract. The kinetic analysis revealed less cooperativity to fructose 6-phosphate (n(H)=1.6) and less inhibition by ATP as compared to the enzyme from baker's yeast. Fructose 2,6-bisphosphate (in micromolar range) and AMP (in millimolar range) were found to overcome ATP inhibition and to increase the affinity to fructose 6-phosphate.