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Purpose: A novel phantom-imaging platform, a set of software tools, for automated and high-precision imaging of the American College of Radiology (ACR) positron emission tomography (PET) phantom for PET/magnetic resonance (PET/MR) and PET/computed tomography (PET/CT) systems is proposed. Methods: The key feature of this platform is the vector graphics design that facilitates the automated measurement of the knife-edge response function and hence image resolution, using composite volume of interest templates in a 0.5 mm resolution grid applied to all inserts of the phantom. Furthermore, the proposed platform enables the generation of an accurate (Formula presented.) -map for PET/MR systems with a robust alignment based on two-stage image registration using specifically designed PET templates. The proposed platform is based on the open-source NiftyPET software package used to generate multiple list-mode data bootstrap realizations and image reconstructions to determine the precision of the two-stage registration and any image-derived statistics. For all the analyses, iterative image reconstruction was employed with and without modeled shift-invariant point spread function and with varying iterations of the ordered subsets expectation maximization (OSEM) algorithm. The impact of the activity outside the field of view (FOV) was assessed using two acquisitions of 30 min each, with and without the activity outside the FOV. Results: The utility of the platform has been demonstrated by providing a standard and an advanced phantom analysis including the estimation of spatial resolution using all cylindrical inserts. In the imaging planes close to the edge of the axial FOV, we observed deterioration in the quantitative accuracy, reduced resolution (FWHM increased by 1–2 mm), reduced contrast, and background uniformity due to the activity outside the FOV. Although it slows convergence, the PSF reconstruction had a positive impact on resolution and contrast recovery, but the degree of improvement depended on the regions. The uncertainty analysis based on bootstrap resampling of raw PET data indicated high precision of the two-stage registration. Conclusions: We demonstrated that phantom imaging using the proposed methodology with the metric of spatial resolution and multiple bootstrap realizations may be helpful in more accurate evaluation of PET systems as well as in facilitating fine tuning for optimal imaging parameters in PET/MR and PET/CT clinical research studies.

Positron emission tomography (PET) imaging of the 18kDa translocator protein (TSPO) has been used to investigate whether microglial activation, an indication of neuroinflammation, is evident in the brain of adults with schizophrenia. Interpretation of these studies is confounded by potential modulatory effects of antipsychotic medication on microglial activity. In the first such study in antipsychotic-free schizophrenia, we have used [11C](R)-PK11195 PET to compare TSPO availability in a predominantly antipsychotic-naïve group of moderate-to-severely symptomatic unmedicated patients (n=8), similarly symptomatic medicated patients with schizophrenia taking risperidone or paliperidone by regular intramuscular injection (n=8), and healthy comparison subjects (n=16). We found no evidence for increased TSPO availability in antipsychotic-free patients compared to healthy controls (mean difference 4%, p=0.981). However, TSPO availability was significantly elevated in medicated patients (mean increase 88%, p=0.032) across prefrontal (dorsolateral, ventrolateral, orbital), anterior cingulate and parietal cortical regions. In the patients with schizophrenia, TSPO availability was also strongly correlated with negative symptoms measured using the Positive and Negative Syndrome Scale across all the brain regions investigated (r=0.651-0.741). We conclude that the pathophysiology of schizophrenia is not associated with microglial activation in the 2-6 year period following diagnosis. The elevation in the medicated patients may be a direct effect of the antipsychotic, although this study cannot exclude treatment resistance and/or longer illness duration as potential explanations. It also remains to be determined whether it is present only in a subset of patients, represents a pro- or anti-inflammatory state, its association with primary negative symptoms, and whether there are significant differences between antipsychotics.

Atmospheric potential gradient measurements at 1 Hz (averaged to 1 minute samples) were taken on the roof of a 6th floor building, approximately 100 m from a busy toll road in Lak Si, northern Bangkok. The measurement period covered the transition from hot season into rainy season. The heavy monsoon rains and heavy traffic can both be significant disruptors of the fair weather atmospheric potential gradient. Heavy traffic increased potential gradient due to aerosol loading, while rain storms increased the magnitude, but also changed the sign of the field due to charge processes within clouds. Increasing relative humidity led to a decrease in potential gradient, implying a lack of hygroscopic growth. Cross correlations of aerosol number in different size fractions showed that aerosols with a D50 value of 155 nm had the strongest correlation. A strong diurnal signal is present in frequency domain analysis of potential gradient.

Positron emission tomography (PET) imaging of the 18 kDa translocator protein (TSPO) has been used to investigate whether microglial activation, an indication of neuroinflammation, is evident in the brain of adults with schizophrenia. Interpretation of these studies is confounded by potential modulatory effects of antipsychotic medication on microglial activity. In the first such study in antipsychotic-free schizophrenia, we have used [

Skeletal muscle and adipose tissues play important roles in maintaining whole-body Glu and N homeostasis by the uptake of Glu and release of Gln. To test the hypothesis that expression of high-affinity Glu transporters (GLAST1, EAAT4, EAAC1, GLT-1) and glutamine synthetase (GS) would increase in longissimus dorsi and adipose tissue of newborn Angus steers randomly assigned ( = 6) to develop through suckling (S; 32 d) and/or weanling (W; 184 d), backgrounding (B; 248 d), and finishing (F; 423 d) production stages. Carcass quality was determined at slaughter to verify shifts in adipose and lean deposition with development. Expression of mRNA (RT-PCR/Southern) and relative protein abundance (Western analysis) were determined in tissue homogenates isolated from longissimus dorsi, and kidney and subcutaneous adipose. The effect of production stage or tissue type on carcass and protein abundance was assessed by 1-way ANOVA using the GLM procedure of SAS, and Fisher's protected LSD procedure was used to separate data means. Neither GLAST1 nor EAAT4 mRNA or protein was detected. EAAC1, GLT-1, and GS mRNA were identified in all tissues, but GLT-1 and GS protein were not detected in kidney or subcutaneous adipose, and GS protein was not detected in longissimus dorsi. The EAAC1 content of subcutaneous ( = 0.06) and kidney ( = 0.02) adipose was 2 times greater in B and F than W steers, whereas GS was 5 times greater (0.07) in B than F steers (B = WF). For longissimus dorsi, EAAC1 (0.01) and GLT-1 (0.04) content decreased with development (SWB = F, S = WB = F, respectively). Within F steers, EAAC1 and GLT-1 mRNA was expressed by subcutaneous, kidney, omental, mesenchymal, and intramuscular adipose tissues, whereas GS mRNA was expressed by all except for intramuscular. Only EAAC1 protein was detected in any adipose tissue, with EAAC1 content being 104% and 112% greater (0.01) in intramuscular than in kidney or subcutaneous adipose, respectively, and not differing (0.45) from omental or mesenchymal adipose. These data demonstrate (1) longissimus dorsi and adipose tissues of steers developing through typical production stages have different capacities for Glu uptake and Gln synthesis, (2) the importance of EAAC1 and GS in adipose metabolism, and (3) the differential metabolic fate of Glu by adipose tissues as steers developed, as evidenced by the marked decrease of GS content in subcutaneous and kidney adipose of F steers.

Twenty-four Angus steers (365 kg ± 4) were assigned randomly to a 2 × 2 factorial arrangement of treatments of either 0 or 350 mg chlortetracycline d-1, with or without Synovex-S to test the effects of chlortetracycline (CTC) and estrogenic implant on release of growth hormone (GH) and thyroid hormones. Steers received ad libitum a concentrate-forage diet over a 112-d feeding study, and growth and carcass characteristics were determined. On days 30, 56, and 106, steers received an injection, via jugular catheter, of 1.0 µg kg-1 BW thyrotropin-releasing hormone (TRH) + 0.1 µg kg-1 BW GH-releasing hormone (GHRH) and blood was collected from -30 to 360 min post-injection. Overall, compared with non-implanted steers, Synovex-S increased (P ≤ 0.009) rate and efficiency of gain, decreased (P = 0.05) time to peak for GH concentrations, whereas baseline concentrations of thyroid-stimulating hormone (TSH) were increased (P = 0.03). Additionally, the TSH response curve (P ≤ 0.05) and magnitude of triiodothyronine (T3; P = 0.01) response were greater following TRH + GHRH challenge for implanted versus nonimplanted steers. There were no main effects of CTC; however, CTC attenuated (P ≤ 0.05) the effects of implant on T3 release, slaughter weight, and carcass composition. In conclusion, Synovex-S alone increased BW gain and enhanced the response of GH, TSH, and T3 to a TRH + GHRH challenge, whereas CTC alone did not affect rate and composition of gain or release of pituitary and thyroid hormones. However, CTC appeared to mitigate the effects of implant on T3 release and carcass composition. Key words: Growth hormone, thyroid, estrogen, chlortetracycline, bovine, growth

Grazing -infected forages results in a variety of reduced animal performance parameters, collectively known as "fescue toxicosis." The initial, limited evaluations of hepatic mechanisms affected by fescue toxicosis have used transcriptomic expression profiling of experimental phenotypes developed by short-term feeding of concentrated ergot alkaloids or fescue seeds to rodents and steers. To assess the effects of fescue toxicosis in growing cattle using a commercially relevant phenotype, we induced fescue toxicosis in beef steers by summer-long grazing (89 to 105 d) of a single high toxic endophyte-infected tall fescue pasture (HE; 0.746 μg/g ergot alkaloids; 5.7 ha; = 10; BW = 267 ± 14.5 kg) vs. a low toxic endophyte tall fescue-mixed pasture (LE; 0.023 μg/g ergot alkaloids; 5.7 ha; = 9; BW = 266 ± 10.9 kg). High toxic endophyte tall fescue-mixed pasture steers had decreased BW (313 vs. 338 kg) and an increased potential for hepatic gluconeogenesis from AA-derived carbons. To gain a greater perspective into fescue toxicosis-induced hepatic metabolism and identify candidate regulatory mechanisms, the goal of the current research was to examine liver samples for changes in gene (mRNA) expression profiles using a Bovine Affymetrix microarray and selected reverse-transcription PCR and immunoblot analyses. The expression (false discovery rate10%;0.01) of 147 genes was increased (7 to 268%) and that of 227 was decreased (4 to 87%) in livers of HE vs. LE steers. The top (1) functional gene category was cell-mediated immune response (33 genes; ≤ 0.012), (2) canonical cell signaling pathway was primary immunodeficiency signaling (8 genes; ≤ 0.0003), and (3) canonical metabolic pathways were oxidative phosphorylation (5 genes; ≤ 0.016) and purine metabolism (8 genes; ≤ 0.029). High toxic endophyte tall fescue-mixed pasture steers had increased ( ≤ 0.022) expression of genes critical for increased (1) Pro () and Ser () synthesis, (2) shunting of AA carbons into pyruvate () and ATP synthesis (, , , COX4, , and ), and (3) mitochondrial mass (COX4). Targeted reverse-transcribed PCR or immunoblot assays corroborated ( ≤ 0.035) these latter microarray findings for , , , , and COX4. Moreover, network analysis identified glucocorticoid receptor-mediated signaling as the most probable mechanism to coordinate the above findings. These results greatly extend our knowledge of the consequences of summer-long grazing of endophyte-infected tall fescue to the hepatic metabolism of growing steers.

Aged beef cows (≥ 8 yr of age) produce calves with lower birth and weaning weights. In mammals, aging is associated with reduced hepatic expression of glutamine synthetase (GS) and alanine transaminase (ALT), thus impaired hepatic Gln-Glu cycle function. To determine if the relative protein content of GS, ALT, aspartate transaminase (AST), glutamate transporters (EAAC1, GLT-1), and their regulating protein (GTRAP3-18) differed in biopsied liver tissue of (a) aged vs. young (3 to 4 yr old) nonlactating, nongestating Angus cows (Exp. 1 and 2) and (b) aged mixed-breed cows with and without COMPUDOSE (17β-estradiol) ear implants (Exp. 3), Western blot analyses were performed. In Exp. 1, 12 young (3.62 ± 0.01 yr) and 13 aged (10.08 ± 0.42 yr) cows grazed the same mixed forage for 42 d (August-October). In Exp. 2, 12 young (3.36 ± 0.01 yr) and 12 aged (10.38 ± 0.47 yr) cows were individually fed (1.03% of BW) a corn-silage-based diet to maintain BW for 20 d. For both Exp. 1 and 2, the effect of cow age was assessed by ANOVA using the MIXED procedure of SAS. Cow BW did not change ( ≥ 0.17). Hepatic ALT (78% and 61%) and GS (52% and 71%) protein content (Exp. 1 and 2, respectively) was decreased ( ≤ 0.01), whereas GTRAP3-18 (an inhibitor of EAAC1 activity) increased ( ≤ 0.01; 170% and 136%) and AST, GLT-1, and EAAC1 contents did not differ ( ≥ 0.17) in aged vs. young cows. In Exp. 2, free concentrations (nmol/g) of Glu, Ala, Gln, Arg, and Orn in liver homogenates were determined. Aged cows tended to have less ( = 0.10) free Gln (15.0%) than young cows, whereas other AA concentrations did not differ ( 0.26). In Exp. 3, 14 aged (10 yr) cows were randomly allotted ( = 7) to sham or COMPUDOSE (25.7 mg of 17β-estradiol) implant treatment (TRT), and had ad libitum access to alfalfa hay for 28 d. Blood and liver biopsies were collected 14 and 28 d after implant treatment. Treatment, time after implant (DAY), and TRT × DAY effects were assessed by ANOVA using the MIXED procedure of SAS. Cow BW was not affected ( ≥ 0.96). Implant increased ( ≤ 0.02) total plasma estradiol by 220% (5.07 vs. 1.58 pg/mL) and GS protein by 300%, whereas the relative content of other proteins was not altered ( ≥ 0.16). We conclude that hepatic expression of ALT and GS are reduced in aged vs. young cows, and administration of 17β-estradiol to aged cows increases plasma estradiol and hepatic GS, but not that of other proteins that support hepatic Glu metabolism.

Meeting Programme Prof David O'Hagan [University of St Andrews, UK]—Enzymatic C–18F Bond Synthesis: A New Strategy for PET Synthesis. Dr Michael Carroll [University of Newcastle, UK]—Studies Towards 6-[18F]Fluoro-m-tyramine using Iodonium Salts. Dr John Gardiner [University of Manchester, UK]—Methods Towards Isotopomer-Versatile Synthesis of 13C-Labelled Carbohydrates. Prof William Lockley [University of Surrey, UK]—Some New Catalytic Systems for Isotope-Exchange Labelling. Dr David Rustidge [Scynexis Europe, UK]—14C-Synthesis—Not Always as Easy as it Looks. Dr Richard Hartley [University of Glasgow, UK]—The Potential for using Titanium Alkylidene Chemistry in Solid-Phase Radiochemical Synthesis. Dr Ian Wilson [Turku Imanet, Finland]—Transition of PET Tracers from Clinical Research Tools to Commercial Diagnostics. Dr Simon Harwood [GlaxoSmithKline, UK]—The Synthesis of Stable Labelled Ketamine. Dr John Herbert [Sanofi-Aventis, UK]—Towards Robust Conditions for Iridium-mediated Exchange. Dr Conor Barry [University of Bristol, UK]—Total Synthesis of Clavosolide A.

Silicification and illitization due to diagenesis result in the modification of porosity and permeability in many North Sea reservoirs, including Greater Alwyn. A good estimation of the mineral composition is required as input/output data for geochemical modelling modes intended to reproduce these effects. Using sedimentological studies to determine laterally equivalent facies, three facies were chosen. ‘Initial states’ for modelling were thus defined as the present-day composition of less-evolved laterally-equivalent facies, ‘final states’ as the present-day composition in the most evolved part of the basin. Precise actual mineral compositions were calculated using a reconciliation program, DATREC. Average compositions calculated for each facies in each field provided data useful in constraining input/output values used in modelling, and gave insights into depositional variation. Arithmetic tests assuming isochemical transformations, constraining the chemical compositions of ‘initial states’ to fit actual mineralogical compositions of ‘final states’, were run using this software. They show that, from an arithmetical point of view, if the assumption of equivalent facies holds, isochemical reactions can occur in some, but not in all facies, thereby demonstrating the importance of checking equivalence of facies and diagenesis hypotheses by quantitative calculations.

The absorption of carnosine, methionine, and methionylglycine (using 35S-methionine and 35S-methionylglycine as representative markers) across ruminal and omasal epithelia collected from four (carnosine) and seven sheep (methionine and methionylglycine) were studied using parabiotic chambers that were repeatedly sampled over a 240-min incubation. The quantity of all substrates transferred was linearly (P < .01) dependent on initial substrate concentration and time. More (P < .01) carnosine, methionine, and methionylglycine was transferred across omasal than across ruminal epithelia. Carnosine was absorbed across both tissues without hydrolysis. Methionylglycine was transferred intact across both tissues. A greater (P < .01) quantity was hydrolyzed by omasal than by ruminal epithelia, after 240 min of incubation. Greater (P < .06) quantities of methionine and methionylglycine accumulated in ruminal tissue after 240 min. Total absorption of methionine and methionylglycine did not differ within tissues, but total absorption of both substrates was greater (P < .01) by omasal tissues. There was little evidence for saturable absorption. These results indicate that omasal epithelial tissue possesses a greater ability to absorb both free and peptide-bound amino acids than does ruminal epithelial tissues.

While isolating siroheme from enzymes or whole cells of Desulfovibrio species, it was discovered that the main product after metal removal and esterification was not the octamethyl ester derivative of sirohydrochlorin, but a monoamide, heptamethyl ester derivative. The structure of this derivative was established by mass spectrometry and NMR. Nuclear Overhauser enhancement measurements in combination with chemical shift analogy arguments indicate that the 2(1)-acetate has been stereospecifically amidated. Other cellular sources of siroheme were investigated, but only the octamethyl ester derivative was found, with no traces of the amide derivative. The results suggest that, in Desulfovibrio, the physiologically active prosthetic group may be an amidated form of siroheme.

This study aimed to derive accurate estimates of regional cerebral blood flow (rCBF) from noisy dynamic [¹⁵O]H₂O PET images acquired on the high-resolution research tomograph, while retaining as much as possible the high spatial resolution of this brain scanner (2-3 mm) in parametric maps of rCBF. The PET autoradiographic method and generalized linear least-squares (GLLS), with fixed or extended to include spatially variable estimates of the dispersion of the measured input function, were compared to nonlinear least-squares (NLLS) for rCBF estimation. Six healthy volunteers underwent two [¹⁵O]H₂O PET scans with continuous arterial blood sampling. rCBF estimates were obtained from three image reconstruction methods (one analytic and two iterative, of which one includes a resolution model) to which a range of post-reconstruction filters (3D Gaussian: 2, 4 and 6 mm FWHM) were applied. The optimal injected activity was estimated to be around 11 MBq kg⁻¹ (800 MBq) by extrapolation of patient-specific noise equivalent count rates. Whole-brain rCBF values were found to be relatively insensitive to the method of reconstruction and rCBF quantification. The grey and white matter rCBF for analytic reconstruction and NLLS were 0.44 ± 0.03 and 0.15 ± 0.03 mL min⁻¹ cm⁻³, respectively, in agreement with literature values. Similar values were obtained from the other methods. For generation of parametric images using GLLS or the autoradiographic method, a filter of ≥ 4 mm was required in order to suppress noise in the PET images which otherwise produced large biases in the rCBF estimates.

Petrographic, X-ray diffraction, and microprobe analyses have been used to assess the significance of illite K-Ar ages from sandstones of two North Sea wells. Three closely spaced samples in one well from the upper Statfjord Formation yield similar ages (69-79 Ma) although the illites formed from different precursor minerals. Pore-filling illite in the upper Brent and the Upper Skagerrak Formations from a second well formed by replacing groundmass clays and other detrital minerals. The average layer charge and K+ content increase slightly with depth (0.69-0.80 K+) due to minor reaction and crystal growth during burial diagenesis. These K-Ar ages increase from 15 to 33 Ma within a 500 m depth interval. The K-Ar age vs. depth relationship for these samples corresponds to the burial rate during the middle Tertiary. In examples of extensive illitization of pore-filling clays in sandstones with little subsequent evolution of the clay minerals, the K-Ar ages indicate the age of diagenetic events.In contrast, illitic minerals in shales from the Skagerrak Formation in the second well yield an age (108 Ma) that is much older than the clays in the sandstones, but is still younger than stratigraphic age. The K-Ar ages from illitic clay in shales reported in the literature can get younger, older, or remain essentially unchanged with increasing depth. These age vs. depth trends reflect the complex interplay of crystal growth and dissolution during diagenesis, as well as probable contamination by non-recrystallized detrital illites. The K-Ar ages of illitic clays, therefore, evolve in a different manner in shales than in sandstones.

After a brief history of ergot alkaloids and ergotism, this review focuses on the metabolism and mechanisms of action of the ergot alkaloids. The authors provide models of how these alkaloids afflict grazing livestock under complex animal-plant/endophyte-environmental interactions. Alkaloid chemistry is presented to orient the reader to the structure-function relationships that are known to exist. Where appropriate, the medical literature is used to aid interpretation of livestock research and to provide insight into potential modes of action and alkaloid metabolism where these are not known for livestock. In closing the paper, we discuss management of ergot alkaloid intoxication in livestock and future research needs for this field of study.

(2010). Potential risk of patient misclassification using a point-of-care testing kit for urine drugs of abuse. British Journal of Biomedical Science: Vol. 67, No. 4, pp. 218-220.

Considerable evidence has been accumulated regarding the absorption of dipeptides and tripeptides, yet, even with the growing body of knowledge, the nutritional and metabolic significance of peptide absorption is not fully understood, especially in ruminants. Muscle, mammary gland, liver, kidney, intestinal mucosa, and other tissues either have been shown to have, or are suspected to have, the ability to utilize peptides as a source of AA to meet cellular demands. Investigations suggest that ruminal microbes have the ability to produce substantial amounts of small peptides as a consequence of their hydrolysis of dietary proteins. The extent to which intact peptides may be absorbed into the blood is controversial. Some of the inconsistency in reported observations may be because of limitations of analytical procedures, species differences, or both. Peptide absorption appears to be an important physiological process in ruminants and may constitute the primary source of absorbed AA. The recent observation that the stomach region of the gastrointestinal tract may be an important site of peptide absorption is highly significant. Emerging evidence for the contribution that peptide absorption makes to AA provisioning of ruminants may change some of the currently held views about protein utilization in these unique animals.

In mammals, the absorption of monosaccharides from small intestinal lumen involves at least 3 sugar transporters (SugT): sodium-dependent glucose transporter 1 (SGLT1; gene SLC5A1) transports glucose and galactose, whereas glucose transporter (GLUT) 5 (GLUT5; gene SLC2A5) transports fructose, across the apical membrane of enterocytes. In contrast, GLUT2 (gene SLC2A2) transports all of these sugars across basolateral and apical membranes. To compare the distribution patterns and sensitivity with nutritional regulation of these 3 SugT mRNA in beef cattle small intestinal tissue, 18 ruminally and abomasally catheterized Angus steers (BW approximately 260 kg) were assigned to water (control), ruminal cornstarch (partially hydrolyzed by alpha-amylase; SH), or abomasal SH infusion treatments (n = 6) and fed an alfalfa-cube-based diet at 1.3 x NE(m) requirement. The SH infusions amounted to 20% of ME intake. After 14- or 16-d of infusion, steers were killed; duodenal, jejunal, and ileal epithelia harvested; and total RNA extracted. The relative amount of SugT mRNA in epithelia was determined using real-time reverse transcription-PCR quantification methods. Basal expression of GLUT2 and SGLT1 mRNA was greater (P0.09) by jejunal than by duodenal or ileal epithelia, whereas basal content of GLUT5 mRNA was greater (Por = 0.02) by jejunal and duodenal than by ileal epithelia. The content of GLUT5 mRNA in small intestinal epithelia was not affected (Por = 0.16) by either SH infusion treatment. In contrast, GLUT2 and SGLT1 mRNA content in the ileal epithelium was increased (Por = 0.05) by 6.5- and 1.3-fold, respectively, after abomasal SH infusion. Duodenal SGLT1 mRNA content also was increased (P = 0.07) by 64% after ruminal SH infusion. These results demonstrate that the ileum of beef cattle small intestine adapts to an increased luminal supply of glucose by increasing SGLT1 and GLUT2 mRNA content, whereas increased ruminal SH supply results in duodenal upregulation of SGLT1 mRNA content. These adaptive responses of GLUT2 and SGLT1 mRNA to abomasal or ruminal SH infusion suggest that beef cattle can adapt to increase their carbohydrate assimilation through small intestinal epithelia, assuming that altered SugT mRNA contents reflect the altered transport functional capacities.

To determine the effect of metabolic acidosis on expression of L-Gln, L-Glu, and L-Asp metabolizing enzymes and transporters, the relative content of mRNA, protein, or mRNA and protein, of 6 enzymes and 5 transporters was determined by real-time reverse transcription-PCR and immunoblot analyses in homogenates of kidney, skeletal muscle, and liver of growing lambs fed a common diet supplemented with canola meal (control; n = 5) or HCl-treated canola meal (acidosis; n = 5). Acidotic sheep had a 790% greater (P = 0.050) expression of renal Na(+)-coupled neutral AA transporter 3 mRNA and a decreased expression of renal glutamine synthetase mRNA (47% reduction, P = 0.037) and protein (57% reduction, P = 0.015) than control sheep. No change in renal cytosolic phosphoenolpyruvate carboxykinase (protein and mRNA), glutaminase (mRNA), or L-Glu dehydrogenase (protein) was found. In skeletal muscle, acidotic sheep had 101% more (P = 0.026) aspartate transaminase protein than did control sheep, whereas no change in the content of 3 Na(+)-coupled neutral AA transporters (mRNA) or 2 high-affinity L-Glu transporter proteins was found. In liver, no change in the content of any assessed enzyme or transporter was found. Collectively, these findings suggest that tissue-level responses of sheep to metabolic acidosis are different than for nonruminants. More specifically, these results indicate the potential capacity for metabolism of L-Asp and L-Glu by skeletal muscle, and L-Gln absorption by kidneys, but no change in hepatic expression of L-Gln metabolism, elaborates previous metabolic studies by revealing molecular-level responses to metabolic acidosis in sheep. The reader is cautioned that the metabolic acidosis model employed in this study differs from the increased plasma lactate-induced metabolic acidosis commonly observed in ruminants fed a highly fermentable grain diet.

Provision of AA has shown success in attenuating proteolytic activity in monogastrics suffering from metabolic acidosis. However, it is unknown whether AA supplementation can provide any beneficial effects to ruminants with nutritionally induced metabolic acidosis. The objective of the current study was to examine the effects of glutamine infusion on various protein degradation components across several tissues in sheep with induced metabolic acidosis. Sheep were assigned to a randomized complete block design with 2 x 2 factorial arrangement of treatments (n = 6 sheep/treatment) consisting of a control or acidosis diet, and receiving a saline or L-glutamine infusion. Sheep were fed diets for 10 d and slaughtered on d 11. Liver, kidney, and muscle samples were collected at slaughter and examined for relative messenger RNA (mRNA) expression of ubiquitin, C8, E2, cathepsin L, cathepsin B, caspase-3, and m-calpain, as well as protein expression of ubiquitin. Relative mRNA expression of C8 (P = 0.02), E2 (P = 0.06), and ubiquitin (P = 0.07) was less in kidney in acidotic vs. control sheep. Additionally, mRNA expression of m-calpain in kidney was greater (P = 0.01) as a result of glutamine infusion. There were no significant alterations (P0.10) in mRNA of any component as a result of acidosis in the liver or muscle. This study demonstrates the inability of metabolic acidosis to increase expression of the ubiquitin-mediated proteolytic pathway in skeletal muscle; however, downregulation of renal mRNA expression of these components is apparent during the induction of metabolic acidosis.

Core KM-3 at Searles Lake, California, comprises 693.4 m of lacustrine sediments deposited over the past 3.2 m.y. The lake water evolved from moderately saline, slightly alkaline, and dominated by Na, Ca, C1, SO4, and HCO3 + CO3 to a highly alkaline brine dominated by Na and CO3 ions. Sediments are chiefly muds and evaporites. Montmorillonite and illite are the principal detrital clay minerals supplied to Searles Lake at present and probably during the late Pliocene and Pleistocene. The drill core is divided into three diagenetic zones on the basis of clay-mineral reactions. The upper zone (0-291.1 m) contains authigenic Fe-illite, Mg-smectite, K-feldspar, and analcime, which average 60-70% of the

This study characterized the effects of nutritionally induced metabolic acidosis with or without Gln infusion on acid-base balance, plasma AA, and plasma NEFA in sheep. In a randomized complete block design with a 2 x 2 factorial arrangement of treatments, 24 fully fleeced sheep (Rideau-Arcott, 63.6 +/- 5.9 kg of BW) were fed a control supplement (CS; 300 g/d of canola meal) or an acidosis supplement (AS; 300 g/d of NutriChlor; HCl-treated canola meal), offered twice daily at 0700 and 1100 h. Sheep were infused at 1400 h daily with 0.3 g of L-glutamine per kg of BW or saline via jugular vein catheters for 7 d. The sheep were individually housed and limit-fed a basal diet of dehydrated alfalfa pellets (1.75 kg/d; 90% DM, 22% CP, and 1.2 Mcal of NE(g)/kg on a DM basis) offered twice daily at 1000 and 1300 h. Blood and urine was sampled daily between 1100 and 1130 h, and blood samples were analyzed for hematocrit, plasma pH, gases, strong ions, AA, and NEFA, whereas urine was analyzed for pH. The AS reduced (P0.01) DMI, urine and plasma pH, blood urea, partial pressure of CO(2), strong ion difference, and plasma HCO(3)(-), and increased (P0.01) plasma K(+), Ca(2+), and Cl(-). The AS with saline infusion increased (Por= 0.03) partial pressure of O(2) and plasma glucose, whereas AS with Gln infusion reduced (P0.01) partial pressure of O(2) and plasma glucose. The AS increased (P0.01) plasma lysine and reduced (P0.01) plasma taurine. Glutamine infusion increased (P = 0.04) plasma leucine with the CS treatment but had no effect (P = 0.89) with the AS treatment. Plasma 16:0, 18:2n-6, 18:3n-3, 20:4n-6, and total NEFA were increased and 18:0 was decreased (P0.001) in AS sheep compared with CS sheep. Infusion of Gln decreased (P0.05) 16:0, 18:2n-6, 18:3n-3, 20:4n-6, and total NEFA compared with saline infusion. Plasma cis-9, trans-11 CLA was elevated (P = 0.001) in AS sheep, whereas plasma cis-9, trans-11 CLA, regardless of the diets, was decreased (P0.02) with Gln infusion. These results show that despite some improvement in overall mobilization of NEFA, Gln infusion did not ameliorate the negative responses associated with metabolic acidosis.

Metabolic acidosis is a condition often induced by ruminal acidosis. Identification of the specific proteolytic pathways affected by metabolic acidosis and characterization of AA concentration changes induced by metabolic acidosis in ruminants has yet to be confirmed. The objective of this study was to examine the effect of nutritionally induced metabolic acidosis on lamb plasma AA and tissue variables, including mRNA and protein expression of components of the ubiquitin-mediated proteolytic pathway. Lambs (n = 10) were divided evenly into treatment groups receiving alfalfa pellets supplemented with 1) a control canola meal supplement, or 2) HCl-treated canola meal supplement for a 10-d treatment period. On d 11, lambs were slaughtered and liver, muscle, and kidney samples were collected to determine mRNA expression of components of the ubiquitin-mediated proteolytic pathway and ubiquitin protein expression. Plasma concentrations of serine (P = 0.06), glycine (P = 0.002), and glutamine (P = 0.04) were greater in acidotic lambs compared with control animals, indicating that protein catabolism may be occurring. However, no alteration (P0.1) in messenger RNA expression of the proteasome subunit C8, ubiquitin-conjugating enzyme E2, or ubiquitin or in ubiquitin protein expression were observed. These results suggest that ubiquitin-mediated proteolysis is not the primary pathway of protein degradation in lambs afflicted with metabolic acidosis.

Although dietary supplementation of cationic AA (CAA), especially l-Lys, is known to be essential for optimal growth of beef cattle, the proteins responsible for absorption of CAA by bovine intestinal epithelia have not been described. Cationic AA transporter-1 (CAT-1) is a major intestinal CAA transporter, demonstrating a high-affinity (muM) transport activity for l-Lys in other mammals, and is widely expressed by small intestinal epithelia of nonruminants, but neither sequence nor expression pattern data exist for CAT-1 in cattle. Therefore, the goal of this research was to compare the relative expression (putative) of CAT-1 mRNA by duodenal, jejunal, or ileal small intestinal epithelia across and within commercially relevant beef cattle production and development stages. Twenty-four Angus steers were assigned randomly (n = 6) to 1 of 4 treatments (suckling, weanling, growing, and finishing) after all steers were born. Duodenal, jejunal, and ileal epithelia were scraped, and total RNA was extracted after the steers were killed at 32, 184, 248, or 423 d of age. Average daily gains of the steers did not differ (1.09 +/- 0.05 kg/d) among stages, whereas the small intestinal length relative to BW decreased (P0.01) with steer development. Using standard reverse transcription-PCR cloning techniques, we generated a partial-length bovine CAT-1 complementary DNA (695 bp; GenBank accession no. DQ399522) from jejunal mRNA samples, which possessed 89 and 87% identities to pig and human CAT-1 orthologs, respectively. On the basis of this bovine-specific genetic data, a real-time PCR-based assay of reverse-transcribed mRNA was developed and used to measure relative changes in bovine CAT-1 mRNA abundance in intestinal epithelia as steers developed. The CAT-1 mRNA was expressed by the duodenum, jejunum, and ileum of all 4 production stages. In contrast to expression by duodenal or ileal epithelium, jejunal expression of CAT-1 mRNA by growing steers was greater (P = 0.005) than that by suckling, weanling, or finishing steers. In terms of the expression of CAT-1 mRNA within production stage, jejunal expression was greater (P = 0.002) than that by duodenum or ileum for growing steers. In contrast, no intestinal site difference was found for suckling, weanling, or finishing steers. These data indicate that previously reported Na(+)-independent uptake of Lys by jejunal and ileal epithelia likely occurred by CAT-1, and that the potential capacity for CAT-1-mediated uptake of CAA for beef steers may be greatest for the "growing" phenotype.

The acid-base status of the extracellular fluid is directly affected by the concentrations of strong basic cations and strong acid anions that are absorbed into the bloodstream from the diet. The objective of this study was to develop and characterize a model for dietary acid challenge in sheep by decreasing the dietary cation-anion difference (DCAD) using NutriChlor (HCl-treated canola meal), an anionic feed supplement. Ten fully fleeced sheep (Rideau-Arcott, 54.3 +/- 6.7 kg of BW) were fed either a control supplement [200 g/d of canola meal, DCAD = 184 mEq/kg of DM, calculated as (Na+ + K+) - (Cl- + S2-)] or an anionic supplement (AS; 200 g/d of NutriChlor, DCAD = -206 mEq/kg of DM) offered twice daily at 0700 and 1100 in a randomized complete block design. The sheep were individually housed and limit-fed a basal diet of dehydrated alfalfa pellets (22% CP and 1.2 Mcal of NE(g)/kg, DM basis) at 1.1 kg of DM/d offered twice daily at 1000 and 1300. Two days before the beginning of the experiment, the sheep were fitted with vinyl catheters (0.86-mm i.d., 1.32-mm o.d.) in the left jugular vein to facilitate blood sampling. Blood and urine samples were obtained daily from 1100 to 1130 on d 1 through 9 and at 0700, 1000, 1300, 1600, and 1900 on d 10. Blood was analyzed for hematocrit, plasma pH, gases, strong ions, and total protein. Urine samples were analyzed for pH. The AS induced a nonrespiratory acid-base disturbance associated with lower (P0.05) plasma pH (7.47 vs. 7.39), lower (P0.05) urine pH (8.13 vs. 6.09), and lower (P0.05) strong ion difference (42.5 vs. 39.5). The AS reduced (P0.05) the concentration of plasma glucose, base excess, and bicarbonate and increased (P0.05) the concentration of K+ and Cl-. Lowering DCAD increased (P0.05) Ca2+ concentrations in plasma by 13%. In conclusion, this dietary model successfully induced a significant acid-base disturbance in sheep. Although the acidifying effects of negative DCAD in the diet may have short-term prophylactic effects of elevating the concentration of Ca2+ in plasma, negative DCAD may have detrimental effects on acid-base balance.

Most animals adapt readily to increased supplies of carbohydrate in the intestinal lumen by increasing enzymes for degradation and increasing glucose transporter activity. However, the extent of upregulation of Na+-dependent glucose cotransporter 1 (SGLT1) activity and content in response to increased delivery of carbohydrate to the small intestinal lumen of ruminants is unclear. Therefore, an experiment was conducted to determine the effect of glucose and starch hydrolysate on the activity and abundance of SGLT1 in the small intestine of steers. In a randomized complete block design, 40 crossbred beef steers (243+/-2 kg BW) were fed 0.163 Mcal of ME/(kg BW0.75(d; W), 0.215 Mcal of ME/(kg BW0.75 x d; 2M), or 0.163 Mcal ME/(kg BW0.75 x d) and infused for 35 d into the rumen (R) or abomasum (A) with 12.6 g/(kg BW0.75 x d) of starch hydrolysate (S) or into the abomasum with 14.4 g/(kg BW0.75 x d) of glucose (G). Steers were slaughtered, and brush-border membrane vesicles were prepared from the small intestinal samples obtained from five equidistant sites along the intestine. Maltase activity in vesicles and homogenates differed with intestinal sampling site (quadratic, P0.001). Steers on the AG treatment yielded a greater intestinal maltase activity (38 nmol glucose x mg protein(-1) x min(-1)) compared with the AS, RS, W, or 2M treatments (34, 26, 23, and 23 nmol glucose x mg protein(-1) x min(-1) respectively [SEM = 3; P = 0.02]). Sodium-dependent glucose uptake averaged 18.4+/-3.94 pmol glucose/(mg protein x s) and was not affected by treatment, but uptake decreased distally along the intestine (P0.001). There was no effect of treatment on SGLT1 protein abundance, but SGLT1 protein abundance increased linearly from the duodenum to the ileum (P = 0.05). The inverse relationship between glucose uptake and SGLT1 abundance suggests that the regulation of brush border Na+-dependent glucose transport capacity is complex, involving factors other than the presence of luminal carbohydrate.

Eight Angus steers (290 +/- 8 kg), surgically prepared with pancreatic pouch-duodenal reentrant cannulas and abomasal infusion catheters were used in a replicated 4 x 4 Latin square experiment to investigate the effects of abomasal infusion of starch hydrolyzate (SH) and/or casein on pancreatic exocrine secretion and plasma concentration of hormones. Steers were fed a basal diet of alfalfa (1.2 x NEm) in 12 equal portions daily. Abomasal infusion treatments (6-L total volume infused per day) were water (control), SH [2.7 g/(kg BW x d)], casein [0.6 g/(kg BW x d)], and SH + casein. Periods were 3 d for adaptation and 8 d of full infusion. Pancreatic juice and jugular blood samples were collected over 30-min intervals for 6 h on d 11. Weight and pH of pancreatic samples were measured, and a 10% subsample was composited and frozen until analysis of total protein and pancreatic enzyme activities. The remaining sample was returned to the duodenum. Plasma was harvested and frozen until analyzed. Pancreatic juice (67 mL/h) and protein (1.8 g/h) secretion rates were not affected by nutrient infusion. There were SH x casein interactions for all pancreatic enzyme secretions (U/h; alpha-amylase, P0.03; trypsin, P0.08; and chymotrypsin, P0.03) and plasma insulin concentration (P0.10). Secretion of pancreatic enzymes was increased by SH (trypsin) and casein (alpha-amylase, trypsin, and chymotrypsin) but not when SH + casein were infused together. Glucose (P0.10) and cholecystokinin octapeptide concentrations (CCK-8; P0.05) were increased by SH, but glucagon was decreased (P0.10). Casein decreased (P0.10) plasma CCK-8 concentrations. These data indicate that positive effects of postruminal casein on enzyme secretion were inhibited by SH, emphasizing the complexity of the regulatory mechanisms involved in dietary adaptation of pancreatic exocrine secretion. Changes in hormone concentration may not relate directly to changes in enzyme secretion.

Our objectives were to determine the effects of neuroendocrine challenge and substrates on in vitro alpha-amylase and trypsin release in pancreatic tissue collected from Holstein calves (n = 24; 88 +/- 3 kg) abomasally infused for 10 d with tap water (control), partially hydrolyzed starch (SH; 4 g/[kg of BW x d]) and/ or casein (0.6 g/[kg of BW x d]). The caudal portion of the pancreas was removed, rinsed with ice-cold saline, cut into approximately 2 x 2-mm segments, and incubated in oxygenated Krebs Ringer bicarbonate buffer containing no substrate (control), glucose, amino acids, or VFA at 39 degrees C. After 60 min of incubation, neurohormonal mimics (none; control), carbachol (acetylcholine analog; 10 microM final), or caerulein (cholecystokinin mimic; 100 nM final) were added to the flasks and tissue was incubated for 60 min. Pancreatic tissue concentrations and in vitro release of alpha-amylase and trypsin decreased (P0.001) in calves abomasally infused with SH. Carbachol increased (P0.10) alpha-amylase and trypsin release in tissue collected from all calves. An effect of caerulein to increase alpha-amylase release (P0.10) was only observed with prior exposure to abomasal casein infusion in vivo or with simultaneous incubation with amino acids in vitro. Caerulein increased (P0.10) trypsin release in tissue collected from all calves except for those receiving SH + casein. Glucose decreased (P0.10) alpha-amylase release from pancreatic tissue collected from calves receiving abomasal control and casein treatments. Amino acids decreased (P0.10) alpha-amylase and trypsin release from pancreatic tissue collected from calves receiving the abomasal control treatment. Glucose, amino acids, and VFA decreased (P0.10) trypsin release from tissue collected from calves receiving abomasal SH. These data indicate that carbachol can stimulate pancreatic enzyme release in vitro. Caerulein, however, is only effective in stimulating in vitro pancreatic enzyme release in tissue from calves with an increased postruminal protein supply or in tissue incubated with amino acids. The results indicate that postruminal and local nutrients might be important in altering the responsiveness to a neuroendocrine challenge and could be an important regulatory event involved with dietary adaptation in ruminants.

Glutamate is a central metabolite for whole-animal energy and N metabolism. This study tested the hypothesis that ileal epithelium, liver, and kidney content of system X-(AG) glutamate transporters EAAC1 and GLT-1 would be up-regulated to support growth of wethers (30 +/- 1.2 kg) fed a forage-based diet for at least 14 d to gain (2.0 x NEm; n = 9) vs. maintain (1.2 x NEm; n = 9) BW. We have previously demonstrated that two high-affinity glutamate transporters (EAAC1, GLT-1) are expressed by these extensive glutamate metabolizing epithelial tissues. Wethers fed at 2.0 x NEm gained (P0.001; 0.26 kg/d) BW, whereas those fed 1.2 x NEm did not. Although plasma concentrations (microM) of glucose and L- or D-glutamate did not differ, plasma glutamine (precursor of glutamate) and alanine concentrations (transamination product of glutamate) were 28% (P0.007) and 22% (P0.072) greater for growing lambs than nongrowing lambs. In tissues, the concentration of L-glutamate in ileum epithelia and D-glutamate of liver was 49% (P0.015) and 181% (P0.042) greater, respectively, in growing vs. nongrowing animals, whereas concentrations of glutamate isoforms did not differ in kidney. Paralleling these increased amino acid concentrations, ileal epithelium contained 313% more (P0.038) EAAC1 protein and liver contained 240% more (P0.001) GLT-1 protein, whereas kidney transporter content did not differ between growing and nongrowing wethers. In contrast to increased EAAC1 and GLT-1 protein content in ileal and liver tissue of growing lambs, messenger RNA levels did not differ. These results indicate that the increased capacity for high-affinity glutamate uptake in growing vs. nongrowing lambs is achieved through increased expression of EAAC1 by ileal epithelium and GLT1 by liver, which parallel increased tissue concentrations of glutamate and plasma concentrations of two major interorgan N carriers, glutamine and alanine.

Central to the assessment of variability of pharmacokinetic parameters is knowledge of bias and variability of the measurement technique, preventing observed differences from being ascribed inappropriate significance. This article presents an evaluation of sources of error in the measurement of normal tissue and tumor pharmacokinetics using 18F-labeled 5-fluorouracil (FU) and PET.A standard approach to data acquisition, processing and analysis was developed using a PET scanner, filtered backprojection reconstruction and region of interest analysis. Fourteen tracer 5-[18F]FU patient studies and a phantom study were completed, with 4 of the patient studies repeated 1 wk later. These data allowed evaluation of the overall reproducibility of the technique and the components of measurement variability due to tissue sampling. The effect of reconstruction technique and sampling region size on quantification was assessed using phantom data.All measured radioactivity versus time curves were tissue specific. Week-to-week variability in the area under this curve (representing combined physiological and measurement difference) was -3% to +15% for liver and -9% to -16% for spleen and kidney. Metastasis variability was greatest at -20%. Visual and computer realignment of the second paired study produced similar results. Interobserver effects were small compared to differences between studies.These results confirm the feasibility of using PET as a pharmacokinetic tool for 5-[18F]FU studies. Although overall experimental error (i.e., random variation in data acquisition, processing and analysis) was low, constraints in data interpretation emerged.

This study reports on the biodistribution and metabolism of the 11C-labeled novel antitumor agent N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) (also known as NSC 601316) in rats (plasma and tissues) and humans (plasma). Information on plasma metabolites was uniquely obtained in humans prior to Phase I clinical trial following i.v. injection of [11C]DACA at tracer dose. DACA was labeled in the N-methyl position using no-carrier-added [11C]iodomethane. Rapid high-performance liquid chromatography methods were developed for metabolite analysis of [11C]DACA. The metabolism of [11C]DACA was investigated in patients by plasma sampling. The biodistribution and metabolism of [11C]DACA was investigated in rats by plasma sampling, sacrifice experiments with tissue analyses, and imaging using positron emission tomography scanning. Analysis of human plasma demonstrated rapid and extensive metabolism of [11C]DACA. The levels of [11C]DACA changed from 77 +/- 8% (SD) at 5 min to 25 +/- 5% at 45 min postinjection. Seven radioactive metabolites were observed in human plasma, and one was identified as [11C]DACA-N-oxide. Rapid clearance of 11C radioactivity from rat blood, plasma, and major organs was observed. The half-life of 11C radioactivity clearance in rat blood between 15 and 90 min was calculated to be 3.2 h; the levels of [11C]DACA in rat plasma decreased from 69 +/- 3% (SD) at 2 min to 29 +/- 1.5% at 25 min. The number of radioactive metabolites in rat plasma was the same as in human plasma except that the proportions differed. Again, one metabolite was identified as the [11C]DACA-N-oxide. Analysis of rat tissues showed rapid and extensive metabolism in tissues, particularly liver and kidney; however, [11C]DACA (i.e., the parent compound) was the major radioactive component in the lung, heart, and brain over 40 min. Positron emission tomography scanning using [11C]DACA in the rat showed little retention of 11C radioactivity in major organs with rapid excretion via gut and kidney. The rat data were consistent with animal (mouse and rat) preclinical data obtained with preexisting techniques with longer-lived isotopes. Labeling of potential anticancer drugs with positron-emitting radionuclides and performing in vivo preclinical evaluation at tracer doses in animals and humans prior to Phase I clinical trials provides unique information that could speed up the assessment of the drug and could potentially assist drug development programs. In this example, there was no unexpected interspecies difference in metabolism of DACA that would have alerted us to make a change in the planned Phase I study.

Research from this laboratory has recently demonstrated that the omasal epithelium of sheep is capable of absorbing dipeptides. In order to express proteins potentially responsible for the mediated absorption of small peptides, size-fractionated poly(A) + RNA (RNA) isolated from omasal epithelial tissue of sheep (average BW 67.5 kg) were injected into defolliculated Xenopus laevis oocytes. The ability of oocytes injected with RNA or water to absorb [ 14 C]glycyl-L-sarcosine (Gly-Sar) from media (usually pH 5.5) was compared. After 4 d ( P

Defolliculated Xenopus laevis oocytes were injected with size-fractionated poly(A)+ RNA (RNA) isolated from the jejunal epithelium of growing pigs (average BW 33.8 kg) to identify proteins capable of Na+ -independent amino acid transport. The ability of oocytes to absorb L-lysine (lysine) or L-leucine (leucine) from Na+- free media was quantified in oocytes after injection of RNA fractions or water. Specific RNA fractions were identified that induced saturable uptake of lysine (Kt = 52 microM) and leucine (Kt = 97 microM), whereas endogenous oocyte uptake was not saturable. Induced uptake of .05 mM lysine by oocytes was inhibited (P.05) 68.1% by 5 mM leucine and 38.9% by .2 mM L-cystine (cystine). Induced uptake of .05 mM leucine was inhibited (P.05) 83.1% by 5 mM lysine and 23.2% by .2 mM cystine. Although not significant (P.05), 5 mM L-glutamate (glutamate) quantitatively stimulated the induced uptake of .05 mM lysine by 18.8% and the induced uptake of .05 mM leucine by 60%. To identify mRNA species responsible for this bo,+ transporter-like activity, oocytes were co-injected with the RNA fractions and degenerate DNA oligomers complementary (antisense) to the cloned human kidney bo,+ amino acid transporter, or (as a negative control) with a DNA oligomer complementary to the rabbit intestinal Na+/glucose cotransporter, or with water. Only those oocytes injected with two specific RNA fractions and the antisense DNA oligomer complementary to the bo,+ transporter displayed reduced (P.05) uptake of lysine (45.7, 55.4%) and leucine (44.1, 65.9%). These results indicate that messenger RNA encoding for a protein capable of stimulating the competitive absorption of lysine and leucine is expressed by the jejunal epithelia of growing pigs.

The genetic organization of the nirD locus of Pseudomonas stutzeri ZoBell, necessary for a catalytically active cytochrome cd1 (EC 1.9.3.2), was determined. The locus comprises the unidirectionally transcribed open reading frames nirFDLGH, downstream of nirMC of the nir gene cluster, and immediately upstream of the norCB operon encoding nitric oxide (NO) reductase (EC 1.7.99.7). Notable sequence relatedness was found between NirF and cytochrome cd1 (NirS), within NirDLGH, and between NirM and NirC, suggesting several gene duplication events in this region. The derived NirF protein (391 amino acids, M(r) 43,137) has 23.8% identity (51.1% overall similarity) with NirS, but lacks the N-terminal heme-c-binding domain of NirS. Insertional mutagenesis of the five open reading frames resulted in the loss of respiratory nitrite reductase activity in vivo and in vitro. Mutant strains, when induced with nitrate for denitrification, synthesized a periplasmic cytochrome cd1 lacking heme d1. The defect was caused by the inability of the cell to synthesize heme d1. The nirD locus is proposed to encode a multimeric and multifunctional enzyme complex involved in the synthesis of heme d1. Mutations in nirFDLGH lowered substantially the expression level of norCB. Nir- mutants, unable to generate NO in vivo, provide indirect evidence for an NO sensor and an inducer role of NO for its cognate reductase.

The National Institute on Aging supports a program of investigation of "biomarkers" of aging (biological parameters that change in magnitude with age), some of which may be among the, as yet unknown, causes of Alzheimer's disease. Exposure to neurotoxicants can also produce symptoms similar to Alzheimer's disease and has been proposed as a causative factor. We have examined the role of dietary factors and age on Alzheimer's disease-like neurohistological and behavioral symptoms as well as on the neurochemical effects of treatment with a prototypical neurotoxicant, trimethyltin (TMT). We found that aging greatly increased the susceptibility of hippocampal neurons to TMT-induced neurodegeneration. Limiting the dietary intake of calories, which has been reported to slow the aging process, also reduced the neurotoxic effects of a given dose of TMT. Our current research focuses on the use of neurotoxins to model neurodegenerative conditions of aging in animals. We propose that by identifying an age-amplified biomarker or biomarkers somewhere along the chain from neurotoxic stimulus to neuronal necrosis we may also discover an up- or down-regulated gene, via it's gene product, that contributes to the etiology of human dementias. Our preliminary data suggest that up-regulated synthesis of post-synaptic glutamate receptors of the kainic acid-preferring subtype may be such a biomarker.

The Lens Opacity Meter 701 (Interzeag, Switzerland) is a recently developed instrument for assessing cataract. It measures the degree of scatter of a red light beam by the lens. Fifty patients prior to cataract surgery had measurements made before and after dilatation of the pupil. Inconsistent results were found at pupil diameters of less than 4 mm. It is postulated that this was due to absorption of scattered light by the iris.

A cross sectional study was performed of factors believed to contribute to the contamination of bovine milk sample cultures submitted to the Ithaca Regional Laboratory of the Quality Milk Promotion Services/New York State Mastitis Control. Of 871 samples entered in the study, 137 (15.7%) were contaminated. There were interactions between the sample source (veterinarian vs dairyman), delivery method, and time between sample collection and arrival at the laboratory. If only those samples collected and hand delivered by the dairyman within 1 day of collection were compared to a like subset of samples collected and hand delivered by veterinarians, no statistically significant differences in milk sample contamination rate (MSCR) were found. Samples were delivered to the laboratory by hand, US Postal Service, United Parcel Service, via the New York State College of Veterinary Medicine Diagnostic Laboratory, or Northeast Dairy Herd Improvement Association Courier. The MSCR was only 7.6% for hand delivered samples, while 26% of Postal Service samples were contaminated. These rates differed significantly from other delivery methods (P less than 0.0001). The USPS samples arrived a longer time after sampling than did samples sent by other routes, and time had a significant effect on MSCR (0 to 1 day, 8.9%; greater than 1 day, 25.9%; P less than 0.01). Samples packaged with ice packs sent by routes other than the Postal Service had a lower MSCR than those not packaged with ice packs, but ice packs did not reduce the MSCR for samples sent by the Postal Service.(ABSTRACT TRUNCATED AT 250 WORDS)

In any course of quantum mechanics it is necessary to explain the behaviour of incompatible dynamical variables when measurements are made of them and also the noncommutation of their corresponding operators. At the outset this presents the student with considerable conceptual difficulties since these ideas are quite outside his everyday experience. The authors present a simple model, an electronic device designed to simulate some of the features of incompatible observables, which can play a useful role in aiding the student to digest the mathematics of noncommuting operators and their eigenfunctions as they appear in an introductory course.

The low temperature heat capacity of a 1% alloy of cobalt in copper in magnetic fields up to 50 kOe has been measured as a function of ageing time at 400 degrees C. The existing theory due to Hayes, Hahn and Wohlfarth (1972) based upon measurements in zero magnetic field, suggests that the excess heat capacity over that of pure copper should be of the form Cmag=A+ gamma T where gamma is a function of the applied field. The present experimental results cannot be represented by a function of this kind but are best represented by Cmag= gamma 'T+CSchottky (H,T) where gamma ' is independent of the applied field and CSchottky (H,T) is the heat capacity of a multilevel Schottky system in which the mean number of thermally populated levels increases from 3.25 to 5.40 as the alloy is aged.