Rumen function and digestion parameters associated with differences between sheep in methane emissions when fed chaffed lucerne hay

An indoor experiment involving 10 rumen-cannulated Romney sheep was conducted in May and June 1998 at AgResearch Grasslands, Palmerston North, New Zealand, under restricted feeding conditions. in order to test the hypothesis that animal factors, in particular rumen fractional outflow rate (FOR) and rumen volume, have an influence on the between-sheep variation in methane (CH4) emission. Sheep were fed 2-hourly on chaffed lucerne hay. Following an acclimatization period of 21 days, the experiment lasted 16 days. Energy and nitrogen (N) balances were measured on days 1-6. Cr-EDTA marker was continuously infused into the rumen from day 9 to 16, and rumen contents emptied and sampled on days 13 and 16. Particulate and fluid FOR were estimated using feed lignin and Cr-EDTA, respectively. Daily CH, production was measured by the sulphur hexafluoride tracer technique on days 2, 5, 6, 12 and 15 of the experiment. CH4 production (g/day) was positively correlated with the pool size of organic matter (OM) in the rumen (OM pool, g) (r = 0.84, P = 0.002), OM intake (OMI, g/day) (r = 0.67, P = 0.04), and the rumen fill (g. wet digesta) (r = 0.76, P = 0.01). Multiple regression analysis showed that CH4 production was best predicted (R-2 = 0.88) as a function of OM pool and the molar % of butyrate; however, OM pool alone accounted for a large proportion (R-2 = 0.71) of the variation in CH4 production. CH4 yield (% gross energy intake, % GEI) was negatively correlated with the particulate FOR (%/h) ( r= -0.75, P = 0.01) and buffering capacity of rumen fluid (mmol HCl) (r = -0.72, P = 0.02) but positively correlated with the digestibility of cellulose (r = 0.66, P = 0.04). Multiple regression analysis showed that CH, yield was best predicted as a function of particulate FOR, OMI (g/kg liveweight(0.75)) and the molar % of butyrate (R-2 = 0.88). Particulate FOR alone explained a large proportion (R-2 = 0.57) of the variation in CH4 Yield. Particulate FOR was negatively correlated with rumen fill (r = -0.69, P = 0.03) and digestibility of cellulose (r = -0.65, P = 0.04). These results suggest that sheep with lower rumen particulate FOR (i.e. longer rumen retention times) had larger rumen fills and higher fibre digestibilities and CH4 yields. If rumen particulate FOR is to be used as a tool for CH4 mitigation, the repeatability of its relationship to CH4 emission must be assessed, preferably under grazing conditions

Persistence of differences between sheep in methane emission under generous grazing conditions

Four low and four high methane (CH4) emitters were selected from a flock of 20 Romney sheep on the basis of CH4 production rates per unit of intake, measured at grazing using the sulphur hexafluoride (SF,) tracer technique. Methane emissions from these sheep were monitored at grazing for four periods (P): October, November, January and February 1999/2000. All measurements were carried out on perennial ryegrass/white clover pasture under generous herbage allowance, and the sheep were maintained on similar pastures during non-measurement periods. The tracer technique was used for all the CH4 measurements and feed DM intake was calculated from total faecal collection and estimated DM digestibility. Data for liveweight (LW), gross energy intake (GEI) and CH4 emission were analysed using split-plot analysis of variance. In addition, a between-period rank order correlation analysis was carried out for CH4 emission data. Low CH4 emitters were heavier (P < 0.05) than the high emitters in all the periods, but they did not differ (P < 0.05) in their gross energy intakes (GEL MJ/kg LW0.75). Low and high CH4 emitters consistently maintained their initial rankings in CH4 yield (% GEI) throughout the subsequent periods and the correlation analysis of rank order for CH4 yield showed strong between-period correlation coefficients, although this was weaker in the last period. It is suggested that feeding conditions that maximize feed intake (e.g. generous allowance of good quality pasture under grazing) favour the expression and persistence of between-sheep differences in CH4 yield

Methane emission by alpaca and sheep fed on lucerne hay or grazed on pastures of perennial ryegrass/white clover or birdsfoot trefoil

Based on the knowledge that alpaca (Lama pacos) have a lower fractional outflow rate of feed particles (particulate FOR) from their forestomach than sheep (San Martin 1987), the current study measured methane (CH4) production and other digestion parameters in these species in three successive experiments (1, 2 and 3): Experiment 1, lucerne hay fed indoors; Experiment 2, grazed on perennial ryegrass/white clover pasture (PRG/WC); and Experiment 3, grazed on birdsfoot trefoil (Lotus corniculatits) pasture (Lotus). Six male alpaca and six castrated Romney sheep were simultaneously and successively fed on the forages either ad libitium or at generous herbage allowances (grazing). CH4 production (g/day) (using the sulphur hexafluoride tracer technique), voluntary feed intake (VFI), diet quality, and protozoa counts and volatile fatty acid concentrations in samples of forestomach contents were determined. In addition, feed digestibility, energy and nitrogen (N) balances and microbial N supply from the forestomach (using purine derivatives excretion) were measured in Experiment 1. Diets selected by alpaca were of lower quality than those selected by sheep, and the voluntary gross energy intakes (GEI, MJ) per kg of liveweight(0.75) were consistently lower (P0.05) in their CH4 yields (% GEI) when fed on lucerne hay (5.1 v. 4.7), but alpaca had a higher CH4 yield when fed on PRG/WC (9.4 v. 7.5, P0.05) in diet N partition or microbial N yield, but alpaca had higher (P<0.05) neutral detergent fibre digestibility (0.478 v. 0.461) and lower (P<0.01) urinary energy losses (5.2 v. 5.8 % GEI) than sheep. It is suggested that differences between these species in forestomach particulate FOR might have been the underlying physiological mechanism responsible for the differences in CH4 yield, although the between-species differences in VFI and diet quality also had a major effect on it

Regulation of Carotenoid Composition and Shoot Branching in Arabidopsis by a Chromatin Modifying Histone Methyltransferase, SDG8

Carotenoid pigments are critical for plant survival, and carotenoid composition is tuned to the developmental stage, tissue, and to environmental stimuli. We report the cloning of the CAROTENOID CHLOROPLAST REGULATORY1 (CCR1) gene. The ccr1 mutant has increased shoot branching and altered carotenoid composition, namely, reduced lutein in leaves and accumulation of cis-carotenes in dark-grown seedlings. The CCR1 gene was previously isolated as EARLY FLOWERING IN SHORT DAYS and encodes a histone methyltransferase (SET DOMAIN GROUP 8) that methylates histone H3 on Lys 4 and/or 36 (H3K4 and H3K36). ccr1 plants show reduced trimethyl-H3K4 and increased dimethyl-H3K4 surrounding the CAROTENOID ISOMERASE (CRTISO) translation start site, which correlates with low levels of CRTISO mRNA. Microarrays of ccr1 revealed the downregulation of 85 genes, including CRTISO and genes associated with signaling and development, and upregulation of just 28 genes. The reduction in CRTISO transcript abundance explains the altered carotenoid profile. The changes in shoot branching are additive with more axillary branching mutants, but the altered carotenoid profile may partially affect shoot branching, potentially by perturbed biosynthesis of the carotenoid substrates of strigolactones. These results are consistent with SDG8 regulating shoot meristem activity and carotenoid biosynthesis by modifying the chromatin surrounding key genes, including CRTISO. Thus, the level of lutein, the most abundant carotenoid in higher plants that is critical for photosynthesis and photoprotection, appears to be regulated by a chromatin modifying enzyme in Arabidopsis thaliana

Subtype-selective regulation of IP₃ receptors by thimerosal via cysteine residues within the IP₃-binding core and suppressor domain

IP(3)R (IP(3) [inositol 1,4,5-trisphosphate] receptors) and ryanodine receptors are the most widely expressed intracellular Ca(2+) channels and both are regulated by thiol reagents. In DT40 cells stably expressing single subtypes of mammalian IP(3)R, low concentrations of thimerosal (also known as thiomersal), which oxidizes thiols to form a thiomercurylethyl complex, increased the sensitivity of IP(3)-evoked Ca(2+) release via IP(3)R1 and IP(3)R2, but inhibited IP(3)R3. Activation of IP(3)R is initiated by IP(3) binding to the IBC (IP(3)-binding core; residues 224–604) and proceeds via re-arrangement of an interface between the IBC and SD (suppressor domain; residues 1–223). Thimerosal (100 μM) stimulated IP(3) binding to the isolated NT (N-terminal; residues 1–604) of IP(3)R1 and IP(3)R2, but not to that of IP(3)R3. Binding of a competitive antagonist (heparin) or partial agonist (dimeric-IP(3)) to NT1 was unaffected by thiomersal, suggesting that the effect of thimerosal is specifically related to IP(3)R activation. IP(3) binding to NT1 in which all cysteine residues were replaced by alanine was insensitive to thimerosal, so too were NT1 in which cysteine residues were replaced in either the SD or IBC. This demonstrates that thimerosal interacts directly with cysteine in both the SD and IBC. Chimaeric proteins in which the SD of the IP(3)R was replaced by the structurally related A domain of a ryanodine receptor were functional, but thimerosal inhibited both IP(3) binding to the chimaeric NT and IP(3)-evoked Ca(2+) release from the chimaeric IP(3)R. This is the first systematic analysis of the effects of a thiol reagent on each IP(3)R subtype. We conclude that thimerosal selectively sensitizes IP(3)R1 and IP(3)R2 to IP(3) by modifying cysteine residues within both the SD and IBC and thereby stabilizing an active conformation of the receptor

Continuum of vasodilator stress from rest to contrast medium to adenosine hyperemia for fractional flow reserve assessment

Objectives: This study compared the diagnostic performance with adenosine-derived fractional flow reserve (FFR) ≤0.8 of contrast-based FFR (cFFR), resting distal pressure (Pd)/aortic pressure (Pa), and the instantaneous wave-free ratio (iFR). Background: FFR objectively identifies lesions that benefit from medical therapy versus revascularization. However, FFR requires maximal vasodilation, usually achieved with adenosine. Radiographic contrast injection causes submaximal coronary hyperemia. Therefore, intracoronary contrast could provide an easy and inexpensive tool for predicting FFR. Methods: We recruited patients undergoing routine FFR assessment and made paired, repeated measurements of all physiology metrics (Pd/Pa, iFR, cFFR, and FFR). Contrast medium and dose were per local practice, as was the dose of intracoronary adenosine. Operators were encouraged to perform both intracoronary and intravenous adenosine assessments and a final drift check to assess wire calibration. A central core lab analyzed blinded pressure tracings in a standardized fashion. Results: A total of 763 subjects were enrolled from 12 international centers. Contrast volume was 8 ± 2 ml per measurement, and 8 different contrast media were used. Repeated measurements of each metric showed a bias &lt;0.005, but a lower SD (less variability) for cFFR than resting indexes. Although Pd/Pa and iFR demonstrated equivalent performance against FFR ≤0.8 (78.5% vs. 79.9% accuracy; p = 0.78; area under the receiver-operating characteristic curve: 0.875 vs. 0.881; p = 0.35), cFFR improved both metrics (85.8% accuracy and 0.930 area; p &lt; 0.001 for each) with an optimal binary threshold of 0.83. A hybrid decision-making strategy using cFFR required adenosine less often than when based on either Pd/Pa or iFR. Conclusions: cFFR provides diagnostic performance superior to that of Pd/Pa or iFR for predicting FFR. For clinical scenarios or health care systems in which adenosine is contraindicated or prohibitively expensive, cFFR offers a universal technique to simplify invasive coronary physiological assessments. Yet FFR remains the reference standard for diagnostic certainty as even cFFR reached only ∼85% agreement

Both D- and L-glucose polyphosphates mimic D-myo-inositol 1,4,5-trisphosphate: new synthetic agonists and partial agonists at the Ins(1,4,5)P3 receptor

Chiral sugar derivatives are potential cyclitol surrogates of the Ca2+-mobilizing intracellular messenger d-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]. Six novel polyphosphorylated analogues derived from both d- and l-glucose were synthesized. Binding to Ins(1,4,5)P3 receptors [Ins(1,4,5)P3R] and the ability to release Ca2+ from intracellular stores via type 1 Ins(1,4,5)P3Rs were investigated. β-d-Glucopyranosyl 1,3,4-tris-phosphate, with similar phosphate regiochemistry and stereochemistry to Ins(1,4,5)P3, and α-d-glucopyranosyl 1,3,4-tris-phosphate are full agonists, being equipotent and 23-fold less potent than Ins(1,4,5)P3, respectively, in Ca2+-release assays and similar to Ins(1,4,5)P3 and 15-fold weaker in binding assays. They can be viewed as truncated analogues of adenophostin A and refine understanding of structure-activity relationships for this Ins(1,4,5)P3R agonist. l-Glucose-derived ligands, methyl α-l-glucopyranoside 2,3,6-trisphosphate and methyl α-l-glucopyranoside 2,4,6-trisphosphate, are also active, while their corresponding d-enantiomers, methyl α-d-glucopyranoside 2,3,6-trisphosphate and methyl α-d-glucopyranoside 2,4,6-trisphosphate, are inactive. Interestingly, both l-glucose-derived ligands are partial agonists: they are among the least efficacious agonists of Ins(1,4,5)P3R yet identified, providing new leads for antagonist development

Comparison of different diastolic resting indexes to iFR: are they all equal?

Background: Pressure measurement for the duration of the wave-free period (WFP) is considered essential for resting-state physiological assessment of coronary stenosis severity using the instantaneous wave-free ratio (iFR). Objectives: The aim of this study was to compare other diastolic resting indexes to iFR. Methods: In the population of the VERIFY2 (Pd/Pa vs iFR in an Unselected Population Referred for Invasive Angiography) study, iFR calculated by proprietary software (Volcano Harvest, Volcano Corporation, Rancho Cordova, California) was compared with the ratio of resting distal coronary pressure and aortic pressure during the complete duration of diastole (dPR), 25% to 75% of diastole (dPR25–75), and midpoint of diastole (dPRmid), along with Matlab calculated iFR (iFRmatlab) and iFR-like indexes shortening the length of the WFP by 50 and 100 ms (iFR−50ms and iFR−100ms), respectively. Mutual differences, Spearman correlations, area under the curve values from receiver-operating characteristic analyses, and diagnostic performance with respect to iFR and fractional flow reserve (FFR) were calculated for all indexes. Results: Median iFR in 197 patients with 257 vessels was 0.91 with an interquartile range of 0.87 to 0.95. The mutual differences (± SD) with iFR were 0.006 ± 0.011 (dPR), 0.001 ± 0.007 (dPR25–75), 0.001 ± 0.008 (dPRmid), 0.005 ± 0.009 (iFRmatlab), 0.003 ± 0.008 (iFR−50ms), and 0.001 ± 0.009 (iFR−100ms). Correlations for all indexes with iFR were &gt;0.99 (p &lt; 0.001 for all). Area under the curve values for predicting iFR were &gt;0.99 for all indexes as well. Diagnostic accuracy compared with FFR was 76% to 77% for all indexes including iFR. Conclusions: All diastolic resting indexes tested were identical to iFR, both numerically and with respect to their agreement with FFR. A numerically equal value to iFR can be determined without restriction to the WFP. Cutoff values, guidelines, and clinical recommendations for iFR can therefore be extended to these other indexes. (Pd/Pa vs iFR in an Unselected Population Referred for Invasive Angiography [VERIFY2]; NCT02377310)

Discordance between resting and hyperemic indices of coronary stenosis severity: the VERIFY 2 study (a comparative study of resting coronary pressure gradient, instantaneous wave-free ratio and fractional flow reserve in an unselected population referred for invasive angiography)

Background—Distal coronary to aortic pressure ratio (Pd/Pa) and instantaneous wave-free ratio (iFR) are indices of functional significance of a coronary stenosis measured without hyperemia. It has been suggested that iFR has superior diagnostic accuracy to Pd/Pa when compared with fractional flow reserve (FFR). We hypothesized that in comparison with FFR, revascularization decisions based on either binary cutoff values for iFR and Pd/Pa or hybrid strategies incorporating iFR or Pd/Pa will result in similar levels of disagreement. Methods and Results—This is a prospective study in consecutive patients undergoing FFR for clinical indications using proprietary software to calculate iFR. We measured Pd/Pa, iFR, FFR, and hyperemic iFR. Diagnostic accuracy versus FFR ≤0.80 was calculated using binary cutoff values of ≤0.90 for iFR and ≤0.92 for Pd/Pa, and adenosine zones for iFR of 0.86 to 0.93 and Pd/Pa of 0.87 to 0.94 in the hybrid strategy. One hundred ninety-seven patients with 257 stenoses (mean diameter stenosis 48%) were studied. Using binary cutoffs, diagnostic accuracy was similar for iFR and resting Pd/Pa with misclassification rates of 21% versus 20.2% (P=0.85). In the hybrid analysis, 54% of iFR cases and 53% of Pd/Pa cases were outside the adenosine zone and rates of misclassification were 9.4% versus 11.9% (P=0.55). Conclusions—Binary cutoff values for iFR and Pd/Pa result in misclassification of 1 in 5 lesions. Using a hybrid strategy, approximately half of the patients do not receive adenosine, but 1 in 10 lesions are still misclassified. The use of nonhyperemic indices of stenosis severity cannot be recommended for decision making in the catheterization laboratory. Clinical Trial Registration—URL: http://www.clinicaltrials.gov. Unique identifier: NCT02377310