Single-molecule vibrational spectroscopy and microscopy

Vibrational spectra for a single molecule adsorbed on a solid surface have been obtained with a scanning tunneling microscope (STM). Inelastic electron tunneling spectra for an isolated acetylene (C2H2) molecule adsorbed on the copper (100) surface showed an increase in the tunneling conductance at 358 millivolts, resulting from excitation of the C-H stretch mode. An isotopic shift to 266 millivolts was observed for deuterated acetylene (C2D2). Vibrational microscopy from spatial imaging of the inelastic tunneling channels yielded additional data to further distinguish and characterize the two isotopes. Single-molecule vibrational analysis should lead to better understanding and control of surface chemistry at the atomic level.     

Inducing and viewing the rotational motion of a single molecule

Tunneling electrons from the tip of a scanning tunneling microscope were used to induce and monitor the reversible rotation of single molecules of molecular oxygen among three equivalent orientations on the platinum(111) surface. Detailed studies of the rotation rates indicate a crossover from a single-electron process to a multielectron process below a threshold tunneling voltage. Values for the energy barrier to rotation and the vibrational relaxation rate of the molecule were obtained by comparing the experimental data with a theoretical model. The ability to induce the controlled motion of single molecules enhances our understanding of basic chemical processes on surfaces and may lead to useful single-molecule devices.     

The Influencing Factor of In Vitro Fertilization and Embryonic Transfer in the Domestic Fowl (Gallus domesticus)

This study was conducted to explore the influencing factors of ova in vitro fertilization (IVF) and transfer of the fertilized ova into the oviduct of recipient hens. The efficiency of fertilization was compared using three aspects: (i) the different time of ova collection and transfer, (ii) egg‐laying period of recipient hen; and (iii) semen volume. The following results are observed: 72%, 40% and 0% of ova were found in ovarian sac in 30～40 min, 50～60 min and more than 90 min post‐oviposition, respectively; 20%, 18%, 14% and 5.8% of ova were fertilized with 0.1, 0.2, 0.5 and 1.0 ml semen, respectively; and 33% and 100% of healthy chickens were hatched from fertile ova with 0.1 and 0.5 ml of semen, respectively. All oocytes obtained from ovary and mid‐oviduct were unfertilized. Embryos were transferred into recipient hens 30 min ± 10 min post‐oviposition, and 70% of shelled eggs were produced. There were no eggs produced in the other transfer times. This demonstrated that live chicken can be obtained by IVF of ova collected shortly after oviposition. It was important that the ovum was transferred into the oviduct infundibulum of recipient hens immediately or shortly after oviposition.     

Major earthquakes occur regularly on an isolated plate boundary fault

The scarcity of long geological records of major earthquakes, on different types of faults, makes testing hypotheses of regular versus random or clustered earthquake recurrence behavior difficult. We provide a fault-proximal major earthquake record spanning 8000 years on the strike-slip Alpine Fault in New Zealand. Cyclic stratigraphy at Hokuri Creek suggests that the fault ruptured to the surface 24 times, and event ages yield a 0.33 coefficient of variation in recurrence interval. We associate this near-regular earthquake recurrence with a geometrically simple strike-slip fault, with high slip rate, accommodating a high proportion of plate boundary motion that works in isolation from other faults. We propose that it is valid to apply time-dependent earthquake recurrence models for seismic hazard estimation to similar faults worldwide.     

Effects of 2.0% end-tidal isoflurane with and without atropine prior to dexmedetomidine administration on cardiovascular variables in dogs

The purpose of this study was to determine the cardiovascular effects of 2.0% end‐tidal isoflurane in dogs administered dexmedetomidine (DEX). Using a randomized crossover design and allowing at least 2 weeks between treatments 12 adult hound dogs of either sex weighing 22 ± 1.7 SD kg were anesthetized by face mask administration of either sevoflurane or isoflurane to facilitate instrumentation prior to administration of treatment drugs. Dogs were intubated and instrumented to enable measurement of heart rate (HR), systolic (SAP), mean (MAP) and diastolic (DAP) arterial pressures, mean pulmonary arterial pressure (PAP), pulmonary capillary wedge pressure (PCWP), central venous pressure (CVP), pulmonary arterial temperature (TEMP), and cardiac output (CO) via thermodilution using 5 mL of 5% dextrose, and recording the average of three replicate measurements. Cardiac index (CI) and systemic (SVR) and pulmonary vascular resistances were calculated. Following completion of instrumentation, dogs were allowed to recover for 40 minutes. After collection of baseline data, dogs were administered one of four treatments at T‐10 minutes prior to injection of DEX (500? g M^–2 IM): 1) saline (SAL); 2) atropine [ATR, 0.02 (n = 6) or 0.04 (n = 6) mg kg^–1 IM]; 3) ISO (2.0% end tidal concentration); or 4) ISO + ATR. Cardiovascular data were collected at T‐20 and T‐5 minutes prior to administration of DEX, and at 5, 10 , 20, 30, 40, and 60 min following DEX. Data were analyzed using anova for repeated measures with post‐hoc differences between means identified using Bonferroni's method (p < 0.05). Differences in ATR dose were not found to be significant and thus results for ATR dose groups were pooled. Administration of SAL (dexmedetomidine alone) was associated with decreases in HR and CO and increases in SAP, MAP, DAP, CVP, and SVR. Administration of ATR was associated with an increase in HR and CO compared with SAL. Administration of ISO was associated with an increase in HR and a decrease in SVR, MAP and CVP compared with SAL. Administration of ISO + ATR was associated with effects similar to that of ISO or ATR alone. We conclude that administration of ISO reduces the increase in SVR associated with administration of DEX and does not adversely affect CO.     

Feline B7.1 and B7.2 proteins produced from swinepox virus vectors are natively processed and biologically active: potential for use as nonchemical adjuvants

Costimulatory ligands, B7.1 and B7.2, have been incorporated into viral and DNA vectors as potential nonchemical adjuvants to enhance CTL and humoral immune responses against viral pathogens. In addition, soluble B7 proteins, minus their transmembrane and cytoplasmic domains, have been shown to block the down regulation of T-cell activation through blockade of B7/CTLA-4 interactions in mouse tumor models. Recently, we developed swinepox virus (SPV) vectors for delivery of feline leukemia antigens for vaccine use in cats [Winslow, B.J., Cochran, M.D., Holzenburg, A., Sun, J., Junker, D.E., Collisson, E.W., 2003. Replication and expression of a swinepox virus vector delivering feline leukemia virus Gag and Env to cell lines of swine and feline origin. Virus Res. 98, 1-15]. To explore the use of feline B7.1 and B7.2 ligands as nonchemical adjuvants, SPV vectors containing full-length feline B7.1 and B7.2 ligands were constructed and analyzed. Full-length feline B7.1 and B7.2 produced from SPV vectors were natively processed and costimulated Jurkat cells to produce IL-2, in vitro. In addition, we explored the feasibility of utilizing SPV as a novel expression vector to produce soluble forms of feline B7.1 (sB7.1) and B7.2 (sB7.2) in tissue culture. The transmembrane and cytoplasmic regions of the B7.1 and B7.2 genes were replaced with a poly-histidine tag and purified via a two-step chromatography procedure. Receptor binding and costimulation activity was measured. Although feline sB7.1-his and sB7.2-his proteins bound to the human homolog receptors, CTLA-4 and CD28, both soluble ligands possessed greater affinity for CTLA-4, compared to CD28. However, both retained the ability to partially block CD28-mediated costimulation in vitro. Results from these studies establish the use of SPV as a mammalian expression vector and suggest that full-length-vectored and purified soluble feline B7 ligands may be valuable, nonchemical immune-modulators.     

Effect of ruminal vs postruminal administration of degradable protein on utilization of low-quality forage by beef steers

An experiment was designed to determine the effects of ruminal and postruminal infusions of ruminally degradable protein (casein) on intake and digestion of low-quality hay by beef steers. Twelve ruminally fistulated Angus x Hereford steers (initial BW = 563 kg) were blocked by weight and assigned to one of three treatments: control (C; hay only) or hay plus ruminal (R) or postruminal (P) infusion of 400 g/d of sodium caseinate. The trial consisted of five periods: 1) 10-d adaptation to the hay diet; 2) 7-d measurement of hay intake (without infusions); 3) 10-d adaptation to protein infusion treatments (intake measurements continued); 4) 7-d measurement of hay intake and digestibility (infusions continued); and 5) 3-d ruminal sampling period (infusions continued). Steers were given ad libitum access to tallgrass-prairie hay (3.4% CP, 76.6% NDF) throughout the study. Casein was administered once daily before feeding, either directly into the rumen or via anchored infusion lines into the abomasum. Hay intake was increased by supplementation (P < 0.01). Ruminal infusion elicited a greater (P = 0.04) increase in hay intake than postruminal infusion. Intake tended (P = 0.11) to be lower in period 4 than in period 2 for control steers but was greater in period 4 than in period 2 (P < or = 0.03) for both R and P steers. The increase in intake between periods 2 and 4 was greater for R than for P steers (P = 0.03). Supplementation improved diet OM digestion (P = 0.04) but not NDF digestion (P = 0.18); however, greater relative error for NDF digestion may have limited the ability to elucidate significant treatment effects. There were no differences in either OM digestion (P = 0.42) or NDF digestion (P = 0.35) between R and P steers. Plasma urea N at 0 and 3 h after feeding on the last day of the experiment was lower (P = 0.05) for C than for R and P steers, but no difference (P = 0.48) was evident between R and P steers. Ruminal ammonia N levels also were increased by supplementation (P < 0.01), with a much larger increase for R than for P steers (P < 0.01). Total VFA concentrations were not affected (P = 0.21) by treatment, but R steers exhibited lower proportions of acetate and higher proportions of isobutyrate, valerate, and isovalerate than P steers (P < 0.01). In conclusion, ruminal and postruminal infusion of a degradable protein source improved forage utilization, although the response in forage OM intake and total digestible OM intake was greater for ruminal infusion than for postruminal infusion.     

Impacts of increasing amounts of supplemental soybean meal on intake and digestion by beef steers and performance by beef cows consuming low-quality tallgrass-prairie forage

Two experiments were conducted to evaluate the impacts of increasing levels of supplemental soybean meal (SBM) on intake, digestion, and performance of beef cattle consuming low-quality prairie forage. In Exp. 1, ruminally fistulated beef steers (n = 20; 369 kg) were assigned to one of five treatments: control (forage only) and .08, .16, .33, and .50% BW/d of supplemental SBM (DM basis). Prairie hay (5.3% CP; 49% DIP) was offered for ad libitum consumption. Forage OM intake (FOMI) and total OM intake (TOMI) were increased (cubic, P = .01) by level of supplemental SBM, but FOMI reached a plateau when the daily level of SBM supplementation reached .16% BW. The concomitant rises in TOMI and OM digestibility (quadratic, P = .02) resulted in an increase (cubic, P = .03) in total digestible OM intake (TDOMI). In Exp. 2, spring-calving Hereford x Angus cows (n = 120; BW = 518 kg; body condition [BC] = 5.3) grazing low-quality, tall-grass-prairie forage were assigned to one of three pastures and one of eight treatments. The supplemental SBM (DM basis) was fed at .08, .12, .16, .20, .24, .32, .40, and .48% BW/d from December 2, 1996, until February 10, 1997 (beginning of the calving season). Performance seemed to reach a plateau when cows received SBM at approximately .30% BW/d. Below this level, cows lost approximately .5 unit of BC for every .1% BW decrease in the amount of supplemental SBM fed. Providing supplemental SBM is an effective means of improving forage intake, digestion, and performance of beef cattle consuming low-quality forages.     

Effect of rumen-degradable intake protein supplementation on urea kinetics and microbial use of recycled urea in steers consuming low-quality forage

We evaluated the effect of increasing amounts of rumen-degradable intake protein (DIP) on urea kinetics in steers consuming prairie hay. Ruminally and duodenally fistulated steers (278 kg of BW) were used in a 4 x 4 Latin square and provided ad libitum access to low-quality prairie hay (4.9% CP). The DIP was provided as casein dosed ruminally once daily in amounts of 0, 59, 118, and 177 mg of N/kg of BW daily. Periods were 13 d long, with 7 d for adaptation and 6 d for collection. Steers were in metabolism crates for total collection of urine and feces. Jugular infusion of (15)N(15)N-urea, followed by determination of urinary enrichment of (15)N(15)N-urea and (14)N(15)N-urea was used to determine urea kinetics. Forage and N intake increased (linear, P < 0.001) with increasing DIP. Retention of N was negative (-2.7 g/d) for steers receiving no DIP and increased linearly (P < 0.001; 11.7, 23.0, and 35.2 g/d for 59, 118, and 177 mg of N/kg of BW daily) with DIP. Urea synthesis was 19.9, 24.8, 42.9, and 50.9 g of urea-N/d for 0, 59, 118, and 177 mg of N/kg of BW daily (linear, P = 0.004). Entry of urea into the gut was 98.9, 98.8, 98.6, and 95.9% of production for 0, 59, 118, and 177 mg of N/kg of BW daily, respectively (quadratic, P = 0.003). The amount of urea-N entering the gastrointestinal tract was greatest for 177 mg of N/kg of BW daily (48.6 g of urea-N/d) and decreased (linear, P = 0.005) to 42.4, 24.5, and 19.8 g of urea-N/d for 118, 59, and 0 mg of N/kg of BW daily. Microbial incorporation of recycled urea-N increased linearly (P = 0.02) from 12.3 g of N/d for 0 mg of N/kg of BW daily to 28.9 g of N/d for 177 mg of N/kg of BW daily. Provision of DIP produced the desired and previously observed increase in forage intake while also increasing N retention. The large percentage of urea synthesis that was recycled to the gut (95.9% even when steers received the greatest amount of DIP) points to the remarkable ability of cattle to conserve N when fed a low-protein diet.