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Objective: Measurement of plasma C2 glucose enrichment is cumbersome. Therefore, the plasma C5 glucose-to-(2)H(2)O rather than the plasma C5-to-C2 glucose ratio commonly has been used to measure gluconeogenesis and glycogenolysis during hyperinsulinemic-euglycemic clamps. The validity of this approach is unknown.Research Design and Methods: Ten nondiabetic and 10 diabetic subjects ingested (2)H(2)O the evening before study. The following morning, insulin was infused at a rate of 0.6 mU . kg(-1) . min(-1) and glucose was clamped at approximately 5.3 mmol/l for 5 h. Plasma C5 glucose, C2 glucose, and (2)H(2)O enrichments were measured hourly from 2 h onward.Results: Plasma C2 glucose and plasma (2)H(2)O enrichment were equal in both groups before the clamp, resulting in equivalent estimates of gluconeogenesis and glycogenolysis. In contrast, plasma C2 glucose and plasma C5 glucose enrichments fell throughout the clamp, whereas plasma (2)H(2)O enrichment remained unchanged. Since the C5 glucose concentration and, hence, the C5 glucose-to-(2)H(2)O ratio is influenced by both gluconeogenesis and glucose clearance, whereas the C5-to-C2 glucose ratio is only influenced by gluconeogenesis, the C5 glucose-to-(2)H(2)O ratio overestimated (P < 0.01) gluconeogenesis during the clamp. This resulted in biologically implausible negative (i.e., calculated rates of gluconeogenesis exceeding total endogenous glucose production) rates of glycogenolysis in both the nondiabetic and diabetic subjects.Conclusions: Plasma C5 glucose-to-(2)H(2)O ratio does not provide an accurate assessment of gluconeogenesis in nondiabetic or diabetic subjects during a traditional (i.e., 2-3 h) hyperinsulinemic-euglycemic clamp. The conclusions of studies that have used this approach need to be reevaluated. [ABSTRACT FROM AUTHOR]

Discusses the abstract of the study 'Evaluation of initial implants on performance and carcass quality in feedlot heifers,' presented at the American Society of Animal Science's Midwestern Branch conference on March 17-19, 2003, in Des Moines, Iowa.

Previously, we demonstrated in test and validation cohorts that type I IFN (T1IFN) activity can predict non-response to tumor necrosis factor inhibitors (TNFi) in rheumatoid arthritis (RA). In this study, we examine the biology of non-classical and classical monocytes from RA patients defined by their pre-biologic treatment T1IFN activity. We compared single cell gene expression in purified classical (CL, n = 342) and non-classical (NC, n = 359) monocytes. In our previous work, RA patients who had either high IFNβ/α activity (>1.3) or undetectable T1IFN were likely to have EULAR non-response to TNFi. In this study comparisons were made among patients grouped according to their pre-biologic treatment T1IFN activity as clinically relevant: "T1IFN undetectable (T1IFN ND) or IFNβ/α >1.3" ( n = 9) and "T1IFN detectable but IFNβ/α ≤ 1.3" ( n = 6). In addition, comparisons were made among patients grouped according to their T1IFN activity itself: "T1IFN ND," "T1IFN detected and IFNβ/α ≤ 1.3," and "IFNβ/α >1.3." Major differences in gene expression were apparent in principal component and unsupervised cluster analyses. CL monocytes from the T1IFN ND or IFNβ/α >1.3 group were unlikely to express JAK1 and IFI27 ( p < 0.0001 and p 0.0005, respectively). In NC monocytes from the same group, expression of IFNAR1, IRF1, TNFA, TLR4 ( p ≤ 0.0001 for each) and others was enriched. Interestingly, JAK1 expression was absent in CL and NC monocytes from nine patients. This pattern most strongly associated with the IFNβ/α>1.3 group. Differences in gene expression in monocytes among the groups suggest differential IFN pathway activation in RA patients who are either likely to respond or to have no response to TNFi. Additional transcripts enriched in NC cells of those in the T1IFN ND and IFNβ/α >1.3 groups included MYD88, CD86, IRF1, and IL8. This work could suggest key pathways active in biologically defined groups of patients, and potential therapeutic strategies for those patients unlikely to respond to TNFi., (Copyright © 2020 Wampler Muskardin, Fan, Jin, Jensen, Dorschner, Ghodke-Puranik, Dicke, Vsetecka, Wright, Mason, Persellin, Michet, Davis, Matteson and Niewold.)

Randomized complete block design experiments ( n = 6 experiments) evaluating steroidal implants (all from Merck Animal Health, Madison, NJ) were conducted in large-pen feedlot research facilities between 2015 and 2018 comparing an 80 mg trenbolone acetate (TBA) and 8 mg estradiol-17β (E 2 ) initial implant (Revalor-IH) and reimplanted with 200 mg TBA and 20 mg E 2 (Revalor-200; REPEATED) to a single 80 mg TBA and 8 mg E 2 uncoated; 120 mg TBA and 12 mg E 2 coated implant (Revalor-XH) at arrival (SINGLE) on growth and carcass responses in finishing heifers. Experiments occurred in Nebraska, Oklahoma, Washington, and Texas. Similar arrival processing was used across experiments where 17,675 heifers [initial body weight = 333 kg SEM (4.1)] were enrolled into 180 pens (90 pens per treatment with 65-240 heifers per pen) and fed for 145-222 d. Only REPEATED heifers were removed from their pen at reimplant. Diets contained monensin and tylosin, consisted of ingredients common to each region, and contained greater than 90% concentrate. Ractopamine hydrochloride was fed for a minimum of 28 d prior to harvest. Linear mixed models were used for all analyses; model-adjusted means for each implant group and the corresponding SEM were generated. Distributions of U.S. Department of Agriculture (USDA) quality grade (QG) and yield grade (YG) were analyzed as ordinal outcomes. No differences ( P ≥ 0.11) were detected for any performance parameters except dry matter intake (DMI), where SINGLE had greater ( P = 0.02) DMI (9.48 vs. 9.38 ± 0.127 kg) compared with REPEATED. Heifers implanted with REPEATED had greater ( P ≤ 0.02) hot carcass weight (HCW; 384 vs. 382 ± 2.8 kg), dressing percentage (64.54 vs. 64.22 ± 0.120%), and ribeye area (91.87 vs. 89.55 ± 0.839 cm 2 ) but less ( P ≤ 0.01) rib fat (1.78 vs. 1.83 ± 0.025 cm) and calculated YG (2.82 vs. 2.97 ± 0.040) and similar ( P = 0.74) marbling scores (503 vs. 505 ± 5.2) compared with SINGLE heifers. Distributions of USDA YG and QG were impacted ( P ≤ 0.03) by treatment such that REPEATED had fewer USDA Prime and YG 4 and 5 carcasses. Heifer growth performance did not differ between implant regimens, but HCW and muscling did, perhaps indicating that REPEATED may be suited for grid-based marketing, and SINGLE might be suited for heifers sold on a live basis depending upon market conditions and value-based grid premiums and discounts. However, these decisions are operational dependent and also may be influenced by factors including animal and employee safety, stress on animals, processing facilities, time of year, labor availability, and marketing strategies., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science.)

An extreme-ultraviolet (XUV) double-stage Raman spectrometer is permanently installed as an experimental end-station at the PG1 beamline of the soft X-ray/XUV free-electron laser in Hamburg, FLASH. The monochromator stages are designed according to the Czerny-Turner optical scheme, adapted for the XUV photon energy range, with optical elements installed at grazing-incidence angles. Such an optical scheme along with the usage of off-axis parabolic mirrors for light collimation and focusing allows for aberration-free spectral imaging on the optical axis. Combining the two monochromators in additive dispersion mode allows for reaching high resolution and superior stray light rejection, but puts high demands on the quality of the optical alignment. In order to align the instrument with the highest precision and to quantitatively characterize the instrument performance and thus the quality of the alignment, optical laser interferometry, Hartmann-Shack wavefront-sensing measurements as well as off-line soft X-ray measurements and extensive optical simulations were conducted. In this paper the concept of the alignment scheme and the procedure of the internal optical alignment are presented. Furthermore, results on the imaging quality and resolution of the first monochromator stage are shown.

The structural dynamics of charge-transfer states of nitrogen-ligated copper complexes has been extensively investigated in recent years following the development of pump-probe X-ray techniques. In this study we extend this approach towards copper complexes with sulfur coordination and investigate the influence of charge transfer states on the structure of a dicopper(i) complex with coordination by bridging disulfide ligands and additionally tetramethylguanidine units [Cu I 2 (NSSN) 2 ] 2+ . In order to directly observe and refine the photoinduced structural changes in the solvated complex we applied picosecond pump-probe X-ray absorption spectroscopy (XAS) and wide-angle X-ray scattering (WAXS). Additionally, the ultrafast evolution of the electronic excited states was monitored by femtosecond transient absorption spectroscopy in the UV-Vis probe range. DFT calculations were used to predict molecular geometries and electronic structures of the ground and metal-to-ligand charge transfer states with singlet and triplet spin multiplicities, i.e. S 0 , 1 MLCT and 3 MLCT, respectively. Combining these techniques we elucidate the electronic and structural dynamics of the solvated complex upon photoexcitation to the MLCT states. In particular, femtosecond optical transient spectroscopy reveals three distinct timescales of 650 fs, 10 ps and >100 ps, which were assigned as internal conversion to the ground state (S n → S 0 ), intersystem crossing 1 MLCT → 3 MLCT, and subsequent relaxation of the triplet to the ground state, respectively. Experimental data collected using both X-ray techniques are in agreement with the DFT-predicted structure for the triplet state, where coordination bond lengths change and one of the S-S bridges is cleaved, causing the movement of two halves of the molecule relative to each other. Extended X-ray absorption fine structure spectroscopy resolves changes in Cu-ligand bond lengths with precision on the order of 0.01 Å, whereas WAXS is sensitive to changes in the global shape related to relative movement of parts of the molecule. The results presented herein widen the knowledge on the electronic and structural dynamics of photoexcited copper-sulfur complexes and demonstrate the potential of combining the pump-probe X-ray absorption and scattering for studies on photoinduced structural dynamics in copper-based coordination complexes.

Objective: To determine mechanisms by which pioglitazone and metformin effect hepatic and extra-hepatic insulin action., Research Design and Methods: Thirty-one subjects with type 2 diabetes were randomly assigned to pioglitazone (45 mg) or metformin (2,000 mg) for 4 months., Results: Glucose was clamped before and after therapy at approximately 5 mmol/l, insulin raised to approximately 180 pmol/l, C-peptide suppressed with somatostatin, glucagon replaced at approximately 75 pg/ml, and glycerol maintained at approximately 200 mmol/l to ensure comparable and equal portal concentrations on all occasions. Insulin-induced stimulation of glucose disappearance did not differ before and after treatment with either pioglitazone (23 +/- 3 vs. 24 +/- 2 micromol x kg(-1) x min(-1)) or metformin (22 +/- 2 vs. 24 +/- 3 micromol x kg(-1) x min(-1)). In contrast, pioglitazone enhanced (P < 0.01) insulin-induced suppression of both glucose production (6.0 +/- 1.0 vs. 0.2 +/- 1.6 micromol x kg(-1) x min(-1)) and gluconeogenesis (n = 11; 4.5 +/- 0.9 vs. 0.8 +/- 1.2 micromol x kg(-1) x min(-1)). Metformin did not alter either suppression of glucose production (5.8 +/- 1.0 vs. 5.0 +/- 0.8 micromol x kg(-1) x min(-1)) or gluconeogenesis (n = 9; 3.7 +/- 0.8 vs. 2.6 +/- 0.7 micromol x kg(-1) x min(-1)). Insulin-induced suppression of free fatty acids was greater (P < 0.05) after treatment with pioglitazone (0.14 +/- 0.03 vs. 0.06 +/- 0.01 mmol/l) but unchanged with metformin (0.12 +/- 0.03 vs. 0.15 +/- 0.07 mmol/l)., Conclusions: Thus, relative to metformin, pioglitazone improves hepatic insulin action in people with type 2 diabetes, partly by enhancing insulin-induced suppression of gluconeogenesis. On the other hand, both drugs have comparable effects on insulin-induced stimulation of glucose uptake.

To determine whether the hepatic insulin resistance of obesity and type 2 diabetes is due to impaired insulin-induced suppression of glycogenolysis as well as gluconeogenesis, 10 lean nondiabetic, 10 obese nondiabetic, and 11 obese type 2 diabetic subjects were studied after an overnight fast and during a hyperinsulinemic-euglycemic clamp. Gluconeogenesis and glycogenolysis were measured using the deuterated water method. Before the clamp, when glucose and insulin concentrations differed among the three groups, gluconeogenesis was higher in the diabetic than in the obese nondiabetic subjects (P < 0.05) and glycogenolysis was higher in the diabetic than in the lean nondiabetic subjects (P < 0.05). During the clamp, when glucose and insulin concentrations were matched and glucagon concentrations were suppressed, both glycogenolysis and gluconeogenesis were higher (P < 0.01) in the diabetic versus the obese and lean nondiabetic subjects. Furthermore, glycogenolysis and gluconeogenesis were higher (P < 0.01) in the obese than in the lean nondiabetic subjects. Plasma free fatty acid concentrations correlated (P < 0.001) with glucose production and gluconeogenesis both before and during the clamp and with glycogenolysis during the clamp (P < 0.01). We concluded that defects in the regulation of glycogenolysis as well as gluconeogenesis cause hepatic insulin resistance in obese nondiabetic and type 2 diabetic humans.

To determine the mechanism(s) by which insulin inhibits endogenous glucose production (EGP) in nondiabetic humans, insulin was infused at rates of 0.25, 0.375, or 0.5 mU. kg(-1). min(-1) and glucose was clamped at approximately 5.5 mmol/l. EGP, gluconeogenesis, and uridine-diphosphoglucose (UDP)-glucose flux were measured using [3-(3)H]glucose, deuterated water, and the acetaminophen glucuronide methods, respectively. An increase in insulin from approximately 75 to approximately 100 to approximately 150 pmol/l ( approximately 12.5 to approximately 17 to approximately 25 microU/ml) resulted in progressive (ANOVA; P < 0.02) suppression of EGP (13.1 +/- 1.3 vs. 11.7 +/- 1.03 vs. 6.4 +/- 2.15 micromol x kg(-1) x min(-1)) that was entirely due to a progressive decrease (ANOVA; P < 0.05) in the contribution of glycogenolysis to EGP (4.7 +/- 1.7 vs. 3.4 +/- 1.2 vs. -2.1 +/- 1.3 micro mol x kg(-1) x min(-1)). In contrast, both the contribution of gluconeogenesis to EGP (8.4 +/- 1.0 vs. 8.3 +/- 1.1 vs. 8.5 +/- 1.3 micro mol x kg(-1) x min(-1)) and UDP-glucose flux (5.0 +/- 0.4 vs. 5.0 +/- 0.3 vs. 4.0 +/- 0.5 micro mol x kg(-1) x min(-1)) remained unchanged. The contribution of the direct (extracellular) pathway to UDP-glucose flux was minimal and constant during all insulin infusions. We conclude that higher insulin concentrations are required to suppress the contribution of gluconeogenesis of EGP than are required to suppress the contribution of glycogenolysis to EGP in healthy nondiabetic humans. Since suppression of glycogenolysis occurred without a decrease in UDP-glucose flux, this implies that insulin inhibits EGP, at least in part, by directing glucose-6-phosphate into glycogen rather than through the glucose-6-phosphatase pathway.

Early changes in cholesteryl ester metabolism of the aorta during the regression of naturally occurring atherosclerotic lesions in pigeon aorta by ileal bypass surgery were examined. Three months after surgery, there was a decrease (50%) in the content of cholesteryl esters in the aorta. Increases in the activity of cholesteryl ester hypdrolase in the lysosomal (P less than 0.05) and the supernatant (P less than 0.01) fractions of the aorta also occurred at this time. There were no differences in the activity of cholesteryl ester synthetase and in the plasma levels of cholesterol and triglycerides between the ileal bypass group and the controls. These results suggest that ileal bypass surgery decreases the level of cholesteryl esters in the aorta, probably because of enhanced cholesteryl ester hydrolysis.

The in vitro effects of prostaglandins E1 and F1alpha on the activity of cholesteryl ester synthetase and cholesteryl ester hydrolase activities of the pigeon aorta were examined. It was found that prostaglandin E1 markedly inhibited the cholesteryl ester hydrolase activity in the supernatant fraction and slightly inhibited the cholesteryl ester synthetase activity. Prostaglandin F1alpha, however, modestly stimulated the cholesteryl ester hydrolase activity both in the microsomal and in the supernatant fraction of the aorta. These observations strongly warrant further studies on the role of prostaglandins in atherogenesis.

The effect of short-term (6 months) administration of conjugated equine estrogen (Premarin) on content and composition of the aortic sterols in male shite Carneau pigeons while they were on a cholesterol-free grain diet was investigated. Estrogen treatment resulted in a 38% increase (P less than 0.05) in free sterol concentration, with a 28.8% concomitant decrease (P less than 0.05) in the percent of cholesteryl esters. The total sterol concentration remained unchanged. This finding suggests that estrogens might influence the synthetic or hydrolytic (or both) processes that control the concentration of cholesteryl esters in the aorta. Fatty acid composition of steryl esters did not change significantly. The cholesterol content of plasma showed a mild reduction (14%) whereas the triglycerides increased significantly (30%).

On cholesterol-free diets, the total fecal steroid excretion was significantly lower in atherosclerosis-susceptible Whit Carneau pigeons than in atherosclerosis-resistant Show Racer pigeons, with major differences in the neutral sterol fraction.

Cholesteryl ester hydrolase activity was measured in the microsomal and supernatant fractions of the aorta of atherosclerosis-susceptible White Carneau and atherosclerosis-resistant Show Racer pigeons while on their normal cholesterol-free diets. Enzyme activities from both fractions showed fatty acid specificities for the hydrolysis of different cholesteryl esters in the following decreasing order: Linoleate greater than oleate greater than palmitate. At 9 months of age (the period of lipid accumulation) the microsomal enzyme activity in the Show Racer breed was significantly higher (P less than 0.001) than in the White Carneau breed, while the supernatant enzyme was slightly higher (P less than 0.05) in the White Carneaux at this age. In older birds (3 years of age) these differences in enzyme activities disappeared.