Selective targeting of human TET1 by cyclic peptide inhibitors: Insights from biochemical profiling.

Ten-Eleven Translocation (TET) enzymes are Fe(II)/2OG-dependent oxygenases that play important roles in epigenetic regulation, but selective inhibition of the TETs is an unmet challenge. We describe the profiling of previously identified TET1-binding macrocyclic peptides. TiP1 is established as a potent TET1 inhibitor (IC50 = 0.26 µM) with excellent selectivity over other TETs and 2OG oxygenases. TiP1 alanine scanning reveals the critical roles of Trp10 and Glu11 residues for inhibition of TET isoenzymes. The results highlight the utility of the RaPID method to identify potent enzyme inhibitors with selectivity over closely related paralogues. The structure-activity relationship data generated herein may find utility in the development of chemical probes for the TETs.

How Lazy Are Pet Cats Really? Using Machine Learning and Accelerometry to Get a Glimpse into the Behaviour of Privately Owned Cats in Different Households.

Surprisingly little is known about how the home environment influences the behaviour of pet cats. This study aimed to determine how factors in the home environment (e.g., with or without outdoor access, urban vs. rural, presence of a child) and the season influences the daily behaviour of cats. Using accelerometer data and a validated machine learning model, behaviours including being active, eating, grooming, littering, lying, scratching, sitting, and standing were quantified for 28 pet cats. Generalized estimating equation models were used to determine the effects of different environmental conditions. Increasing cat age was negatively correlated with time spent active (p < 0.05). Cats with outdoor access (n = 18) were less active in winter than in summer (p < 0.05), but no differences were observed between seasons for indoor-only (n = 10) cats. Cats living in rural areas (n = 7) spent more time eating than cats in urban areas (n = 21; p < 0.05). Cats living in single-cat households (n = 12) spent more time lying but less time sitting than cats living in multi-cat households (n = 16; p < 0.05). Cats in households with at least one child (n = 20) spent more time standing in winter (p < 0.05), and more time lying but less time sitting in summer compared to cats in households with no children (n = 8; p < 0.05). This study clearly shows that the home environment has a major impact on cat behaviour.

A Small-Molecule Inhibitor of Factor Inhibiting HIF Binding to a Tyrosine-flip Pocket for the Treatment of Obesity.

In animals limiting oxygen upregulates hypoxia-inducible factor (HIF) promoting a metabolic shift towards glycolysis. Factor inhibiting HIF (FIH) is an asparaginyl hydroxylase that regulates HIF function by reducing its interaction with histone acetyl transferases. HIF levels are negatively regulated by the HIF prolyl hydroxylases (PHDs), which like FIH, are 2-oxoglutarate(2OG) oxygenases. Genetic loss of FIH promotes both glycolysis and aerobic metabolism. FIH has multiple non-HIF substrates making it challenging to connect its biochemistry with physiology. A structure-mechanism guided approach identified a highly potent in vivo active FIH inhibitor, ZG-2291, binding of which promotes a conformational flip of a catalytically important tyrosine, enabling selective inhibition of FIH over other JmjC subfamily 2OG oxygenases. Consistent with genetic studies, ZG-2291 promotes thermogenesis and ameliorates symptoms of obesity and metabolic dysfunction in ob/ob mice. The results reveal ZG-2291 as a useful probe for the physiological functions of FIH and identify FIH inhibition as a promising strategy for obesity treatment.

The Use of Triaxial Accelerometers and Machine Learning Algorithms for Behavioural Identification in Domestic Cats (Felis catus): A Validation Study.

Animal behaviour can be an indicator of health and welfare. Monitoring behaviour through visual observation is labour-intensive and there is a risk of missing infrequent behaviours. Twelve healthy domestic shorthair cats were fitted with triaxial accelerometers mounted on a collar and harness. Over seven days, accelerometer and video footage were collected simultaneously. Identifier variables (n = 32) were calculated from the accelerometer data and summarized into 1 s epochs. Twenty-four behaviours were annotated from the video recordings and aligned with the summarised accelerometer data. Models were created using random forest (RF) and supervised self-organizing map (SOM) machine learning techniques for each mounting location. Multiple modelling rounds were run to select and merge behaviours based on performance values. All models were then tested on a validation accelerometer dataset from the same twelve cats to identify behaviours. The frequency of behaviours was calculated and compared using Dirichlet regression. Despite the SOM models having higher Kappa (>95%) and overall accuracy (>95%) compared with the RF models (64-76% and 70-86%, respectively), the RF models predicted behaviours more consistently between mounting locations. These results indicate that triaxial accelerometers can identify cat specific behaviours.

Practices and opinions of New Zealand sheep farmers towards pain management in lambs during castration and/or tail docking.

AIMS: To collect baseline data from New Zealand sheep farmers on techniques for docking and castrating lambs, their perceptions of the level of pain lambs experience following these procedures, and their opinions about the use of pain relief. METHODS: From a voluntary cross-sectional survey of New Zealand sheep farmers, descriptive statistics were provided for quantitative study variables. Thematic analysis was performed on the free-text comments. Univariable logistic regression was used to assess factors associated with farmers indicating they would consider using a device allowing administration of pain relief simultaneously with castration and tail docking. RESULTS: There were 432 survey responses containing sufficiently complete data for analysis. Of the 340 individuals (77.5%) always or sometimes performing castration of ram lambs, 242 (72.2%) used rubber rings for complete castration, 23 (6.9%) used the short scrotum method for cryptorchid castration, and 75 (22.4%) used a combination of both methods. Of the 423 individuals (97.9%) who indicated that they always or sometimes performed tail docking, 245 (57.9%) used a hot iron only, 148 (35.0%) used a rubber ring only, 26 (5.8%) used both methods, 3 (0.7%) used a surgical knife, and 1 (0.2%) provided no response. Less than 2% of respondents always or sometimes used pain relief for these procedures. Of the 432 respondents, 139 (32.2%) and 180 (41.7%) strongly agreed that castration and tail docking do not cause sufficient pain to warrant using pain relief, respectively. Time and cost were identified as major barriers to providing pain relief. In the unadjusted logistic regression analyses, respondents who were female, had high levels of education, had been farming < 20 years, who believed lambs experienced high levels of pain following the procedures, and who believed pain lasted longer than 6 hours, were more likely to indicate willingness to use pain control devices. CONCLUSION AND CLINICAL RELEVANCE: Our results suggest very few New Zealand sheep farmers currently provide lambs with pain relief following tail docking or castration. This is likely due to the perception that the procedures are not painful enough to warrant pain relief, and concern over time and cost. This highlights the need to educate farmers about lamb pain and distress following tail docking and castration, and its negative impact on animal welfare. Farmers also need pain relief techniques and tools that can be administered simultaneously with these procedures to save time and labour cost.

Ewe wastage in commercial sheep flocks: a review of current knowledge.

Ewe wastage encompasses ewes that do not reach the end of their potential productive lives and is the combination of premature culling and on-farm mortality. Increased ewe wastage results in reduced flock productivity and profitability, primarily driven by the need for increased replacement ewe lambs to maintain flock numbers and the costs associated with these additional replacements (e.g. reduced sale lambs, reduced flock age resulting in lower lamb production, and reduced ability to utilise terminal sires). Annual ewe mortality rates are reported to range from 2.8 to 40.2% in New Zealand commercial flocks, while wastage over a ewe's productive lifespan (i.e. mortality and premature culling combined to 6-7 years-of-age) is reported to be up to 90%. To date, ewe wastage has received relatively little attention in the published literature. This article reviews published studies describing wastage in non-dairy ewes, focusing on New Zealand commercial flocks, although overseas pasture-based data is also considered. The relevance of, and challenges associated with, investigating ewe wastage are discussed. Ewe culling and mortality data is outlined, and causes are described. Known key risk factors for wastage, namely, reproductive performance, udder health, dental health, body condition score and breeding of ewe lambs, are reviewed. Finally, current knowledge gaps and future areas of research are identified and evaluated.

What Are Sheep Doing? Tri-Axial Accelerometer Sensor Data Identify the Diel Activity Pattern of Ewe Lambs on Pasture.

Monitoring activity patterns of animals offers the opportunity to assess individual health and welfare in support of precision livestock farming. The purpose of this study was to use a triaxial accelerometer sensor to determine the diel activity of sheep on pasture. Six Perendale ewe lambs, each fitted with a neck collar mounting a triaxial accelerometer, were filmed during targeted periods of sheep activities: grazing, lying, walking, and standing. The corresponding acceleration data were fitted using a Random Forest algorithm to classify activity (=classifier). This classifier was then applied to accelerometer data from an additional 10 ewe lambs to determine their activity budgets. Each of these was fitted with a neck collar mounting an accelerometer as well as two additional accelerometers placed on a head halter and a body harness over the shoulders of the animal. These were monitored continuously for three days. A classification accuracy of 89.6% was achieved for the grazing, walking and resting activities (i.e., a new class combining lying and standing activity). Triaxial accelerometer data showed that sheep spent 64% (95% CI 55% to 74%) of daylight time grazing, with grazing at night reduced to 14% (95% CI 8% to 20%). Similar activity budgets were achieved from the halter mounted sensors, but not those on a body harness. These results are consistent with previous studies directly observing daily activity of pasture-based sheep and can be applied in a variety of contexts to investigate animal health and welfare metrics e.g., to better understand the impact that young sheep can suffer when carrying even modest burdens of parasitic nematodes.

Behaviour of twin- and triplet-born lambs and their dam 3 to 18 hours after birth is not a useful predictor of lamb survival to weaning.

OBJECTIVE: An experiment was designed to determine if behaviour traits expressed by twinand triplet-bearing lambs and their dams at 3 to 18 hours of age (after the immediate ewelamb bonding had occurred) were associated with lamb survival to weaning. METHODS: The behaviour of twin and triplet lambs and their dams was assessed in the paddock at 3 to 18 hours after birth. Observations were made of the number of high- and low-pitched bleats, time to stand, make contact with dam, suck from dam and follow dam were recorded for each lamb. The maternal behaviour score of each dam was assessed. A random sub-sample of lambs were assessed during a maternal-recognition test at 12 or 24 hours of age. Traits included time spent standing, sitting, walking, time taken to reach the ewes and time spent with the ewes as well as the number of high- and low-pitched bleats emitted by the lamb. RESULTS: In the paddock, for each additional second required for twin-born lambs to follow their dam, lambs were 1.004 (95% confidence interval [CI] 1.000 to 1.008) times more likely to survive to weaning (p<0.05). The opposite relationship, however, was seen in triplet lambs. For each additional second required for triplet-born lambs to follow their dam, lambs were 0.996 (95% CI 0.993 to 0.999) times as likely to survive to weaning (p<0.05). During the maternal recognition test, twin-born lambs were 0.989 (95% CI 0.979 to 1.000) times as likely to survive to weaning for every additional second they took to reach the contact zone (p<0.05). Similarly, triplet-born lambs were 0.994 (95% CI 0.989 to 0.999) as likely to survive for every additional second they took to reach their dam (p<0.05). CONCLUSION: All ewe behaviours and the majority of lamb paddock and test behaviours were not associated with the survival of twin- or triplet-born lambs and, therefore, are of little use as indicators of lamb survival to weaning.

Investigating associations between lamb survival to weaning and dam udder and teat scores.

Aims: To examine a range of udder and teat traits in Romney ewes and to describe the frequency with which different scores occur, and to investigate associations between lamb survival to weaning and ewe udder and teat scores. Methods: Mixed-age, mature Romney ewes (n = 1,009) were enrolled from a commercial sheep flock located in the Wellington region of New Zealand in January 2017. A range of udder and teat traits were scored in all ewes, using visual assessment and palpation, at pre-mating (February), pre-lambing (October), docking (November) and weaning (January 2018). During the lambing period each newborn lamb was matched to its dam, with lamb mortalities recorded until weaning. Associations between udder and teat scores and lamb survival to weaning were examined using multivariable models for each udder-scoring time. Results: Records from 981 ewes and 1,822 live-born lambs were included in analyses, with 252 (13.8%) lambs recorded dead between birth and weaning. Lambs born to ewes with pre-mating udder scores of lump or hard had 4.9 (95% CI = 2.6-9.6, p = 0.003) and 3.0 (95% CI = 1.5-6.1, p < 0.001) increased odds of failure to survive to weaning, respectively, compared with lambs whose dams had normal udder scores. Lambs born to ewes with mastitis at docking or weaning had 3.0 (95% CI = 1.5-5.9, p = 0.001) and 3.9 (95% CI = 1.3-11.6, p = 0.013) increased odds of failure to survive to weaning, respectively, compared with lambs whose dams did not have mastitis. Offspring of dams with asymmetrical udders at docking or weaning had 3.3 (95% CI = 2.2-4.9, p < 0.001) and 2.5 (95% CI = 1.5-4.0, p < 0.001) increased odds of failure to survive, respectively, compared with lambs whose dams had symmetrical udders. Conclusion and Clinical Relevance: Pre-mating udder palpation scores of hard or lump were associated with increased odds of lambs not surviving to weaning compared with normal scores, and could be used to identify ewes that are likely to be unsuitable for retaining in the breeding flock. Farmers could also use clinical mastitis scores and udder symmetry scores at docking or weaning to identify ewes whose lambs had greater odds of failure to survive to weaning. However these scores do not provide an indication of future performance, therefore further investigation into the impact of the present season's score on future seasons' lamb survival is required.

Associations between lamb growth to weaning and dam udder and teat scores.

Aims: To investigate associations between lamb growth to weaning and dam udder and teat scores measured at pre-mating, pre-lambing, docking and weaning. Methods: Mature Romney ewes (n = 1,009) were enrolled from a commercial sheep flock located near Masterton, in the Wellington region of New Zealand in 2017. A range of udder and teat traits were scored in all ewes, using visual assessment and palpation, at pre-mating, pre-lambing, docking and weaning. During the lambing period, each newborn lamb was matched to its dam and lamb sex, birthweight and birth-rank were recorded. A rearing rank was allocated to each live-lamb at weaning, when all lambs were weighed (n = 1,570), allowing calculation of daily growth rates (g/day). Associations between udder and teat scores and lamb growth rates to weaning were examined using multivariable models for each udder-scoring time. Results: Growth rates of lambs whose dams had udder palpation scores of hard, or both teats recorded as abnormal, pre-mating were lower than lambs whose dams had normal scores (229.9 (95% CI = 213.2-246.6) vs. 254.5 (95% CI = 245.6-263.5) g/day; p = 0.011) and (227.4 (95% CI = 208.3-246.6) vs. 247.9 (95% CI = 235.7-260.2) g/day; p = 0.024), respectively. Growth rates of lambs whose dams had clinical mastitis at docking or weaning were lower than those without mastitis (215.8 (95% CI = 199.9-231.7) vs. 235.4 (95% CI = 225.4-255.0) g/day; p = 0.007) and (220.0 (95% CI = 205.2-234.8) vs. 254.7 (95% CI = 248.9-260.5) g/day; p < 0.001), respectively. Growth rates of lambs whose dams had asymmetrical udders at docking or weaning were lower than lambs whose dams had symmetrical udders (204.6 (95% CI = 189.7-219.5) vs. 240.2 (95% CI = 225.4-255.0) g/day; p < 0.001) and (223.3 (95% CI = 213.9-232.7) vs. 242.2 (95% CI = 229.4-255.0) g/day; p = 0.014), respectively. Conclusions and Clinical Relevance: Pre-mating udder palpation and teat palpation scores can be used to identify ewes whose lambs are predicted to have lower growth to weaning. Assuming a mean lamb age at weaning of 84.4 days, lambs born to ewes with a pre-mating score of hard would be expected to have a mean weaning weight that was 2.1 kg less than those whose dams had normal scores. Udder palpation, udder symmetry and clinical mastitis scores during lactation were also associated with lamb growth rates. Abbreviation: CALW: Conceptus-adjusted liveweight.

A comparison of liveweight gain of lambs weaned early onto a herb-clover mixed sward and weaned conventionally onto a ryegrass-clover pasture and herb-clover mixed sward.

OBJECTIVE: The aim of the present study was to identify the impact of early weaning of lambs at approximately seven weeks of age onto a herb-clover mix on the liveweight gain of lambs and their dams. METHODS: In 2015, twin-born lambs that weighed a minimum of 16 kg (n = 134) were randomly allocated to one of three treatments: i) Early-weaned (58 days after the midpoint of lambing) onto an unrestricted allowance (>1,200 kg dry matter/ha) of herb-clover mix (HerbEW); ii) Lambs+dams unweaned onto an unrestricted allowance of herb-clover mix until conventional weaning (95 days after the midpoint of lambing) (HerbCW); iii) Lambs+ dams unweaned onto an unrestricted allowance of grass-clover pasture until conventional weaning (GrassCW). In 2016, twin-born lambs that weighed a minimum of 16 kg (n = 170) were randomly allocated to one of four treatments: i), ii), iii) (similar to 2015) and iv) Lambs+ dams unweaned onto a restricted allowance (<1,200 kg dry matter/ha) of grass-clover pasture until conventional weaning (93 days after the midpoint of lambing) (Restricted-GrassCW). RESULTS: In 2015, liveweight gain from L58 to L95 of HerbCW and GrassCW lambs did not differ (p>0.05), but were greater than HerbEW lambs. In 2016, HerbCW lambs had greater (p<0.05) liveweight gains from L51 to L93 than GrassCW followed by HerbEW and Restricted-GrassCW lambs. In 2015, liveweight gain from L58 to L95 of HerbEW ewes were greater than both GrassCW and HerbCW ewes while in 2016, liveweight gain of from L51 to L93 GrassCW and HerbCW ewes did not differ (p>0.05) but were greater (p<0.05) than those of HerbEW and Restricted-GrassCW ewes. CONCLUSION: These results indicate that when grass-clover pasture supply can be maintained at unrestricted intake level, there may be no benefit of weaning lambs early. However, at restricted pasture conditions lambs can achieve greater liveweight gains when weaned early onto a herb-clover mix.

Associations between liveweight, body condition score and previous reproductive outcomes, and the risk of ewes bred at 18-months of age being dry at docking.

AIMS Firstly, to investigate associations between liveweight and body condition score (BCS) of two-tooth ewes (18-months-old at breeding) at breeding, pregnancy diagnosis (PD) and pre-lambing and the risk of being dry at docking, on commercial New Zealand sheep farms. Secondly, to investigate the association between previous reproductive outcomes as ewe lambs, and risk of being dry at docking as two-tooth ewes. METHODS Two-tooth ewes (n=9,006) were enrolled in four cohorts from three commercial sheep farms between 2010-14. Ewes were weighed and BCS assessed immediately pre-breeding, at PD (mid-pregnancy) and pre-lambing. At PD, ewes were identified as either non-pregnant, or having single or multiple fetuses. Palpation and examination of udders at docking was used to classify each ewe as either lactating or dry at docking. RESULTS Overall, 437/8,025 (5.4%) of ewes that were diagnosed pregnant at PD were dry at docking. The risk of being dry at docking decreased with increasing pre-lambing conceptus adjusted liveweight (CALW) on all farms (p≤0.002); for 2010-born ewes from Farm A the OR=0.87 (95% CI=0.81-0.92); for Farm B the OR=0.88 (95% CI=0.83-0.92) and for Farm C the OR=0.86 (95% CI=0.79-0.95). The risk of being dry at docking also decreased with increasing CALW gain from PD to pre-lambing for all farms (p≤0.003); for 2010-born ewes from Farm A the OR=0.89 (95% CI=0.84-0.94); for Farm B the OR=0.85 (95% CI=0.81-0.89) and for Farm C the OR=0.88 (95% CI=0.80-0.96). There was no association between BCS at breeding, PD or pre-lambing and the risk of being dry at docking for 2010-born ewes from Farm A, Farm B or Farm C (p>0.05). For 2010-born ewes on Farm A, the risk of being dry at docking was greater for two-tooth ewes that were previously dry at docking as ewe lambs than those that were lactating at docking as ewe lambs (OR=1.7 (95% CI=1.1-2.8); p=0.018), but this difference was not observed for ewes on Farm B or Farm C (p>0.5). CONCLUSIONS There were negative associations between ewe CALW pre-lambing, and CALW gain between PD and pre-lambing, and risk of being dry at docking. For all cohorts, heavier ewes and those that gained CALW were less likely to be dry at docking than lighter ewes or those that lost CALW, however these relationships varied between cohorts.

The effect of nutrition and body condition of triplet-bearing ewes during late pregnancy on the behaviour of ewes and lambs.

OBJECTIVE: Triplet-born lambs are less likely to survive to weaning than twin-born or single-born lambs. Appropriate ewe-lamb bonding behaviours and lamb vigour behaviours are necessary for survival of lambs. The aim of this experiment was to determine whether maternal nutrition during late pregnancy influenced behaviour of the ewe and her lambs soon after birth, and to determine whether mid-pregnancy body condition score (BCS) influenced any behavioural response. METHODS: The experiments included ewes that were in BCS 2.0, 2.5, or 3.0 in mid-pregnancy and were fed either ad libitum or to pregnancy-maintenance requirements in late-pregnancy (day 115 until 136 in experiment one, and day 128 until 141 in experiment two). The time taken for lambs to stand, contact dam, suck from dam and follow dam was recorded three to 18 h after birth. The number of high- and low-pitched bleats emitted by the ewe and lambs was recorded, along with maternal behaviour score (MBS) of the ewe. Lambs in experiment two underwent a maternal-recognition test at 12 or 24 h. RESULTS: There were significant effects of feeding treatment on bleating behaviour of ewes and lambs, but these were inconsistent among BCS groups and between experiments. Lamb vigour behaviours were not affected by feeding treatment. In experiment one, there was no effect of feeding treatment or BCS on MBS, but in experiment two, ewes in BCS3 in mid-pregnancy had greater MBS than ewes in BCS2 in mid-pregnancy (MBS 3.1/5 vs MBS 2.1/5; p<0.05). CONCLUSION: Given there were no repeatable effects on behaviour of ewes and lambs, ad libitum feeding rather than feeding for pregnancy-maintenance requirements cannot be used to improve behaviours soon after birth of triplet-bearing ewes in BCS 2-3 and their lambs in extensive pastoral conditions.

Investigation of fetal loss in ewe lambs in relation to liveweight changes and progesterone concentrations in early to mid gestation.

AIMS: To investigate associations between changes in liveweight and fetal loss in ewe lambs and to determine whether fetal loss was associated with concentrations of progesterone in early and mid-gestation. METHODS: Ewe lambs on two commercial sheep farms (Flock A; n=504 and Flock B; n=460) in the southern North Island of New Zealand were weighed, and body condition score was assessed, prior to breeding and throughout pregnancy. Pregnancy diagnosis was undertaken approximately 56 (PD1), 80 (PD2) and 110 (PD3) days after the start of breeding and fetal loss determined. Blood samples were collected from a cohort of 407 ewe lambs between 45-54 and 73-84 days of gestation. Those that subsequently were diagnosed with fetal loss (n=16) were matched with ewe lambs that maintained pregnancy (n=16), and concentrations of progesterone were measured in the plasma samples from these animals. RESULTS: Fetal loss was diagnosed in 55/472 (11.7%) pregnant ewes from Flock A and 14/378 (3.7%) from Flock B. In both flocks the change in conceptus-free liveweight (CFLW) between PD1 and PD2 differed between ewe lambs with and without fetal loss; in Flock A the mean change was -59.9 (95% CI=-80.1 to -39.7) g/day vs. -3.43 (95% CI=-9.8 to 3.0) g/day (p<0.001), and in Flock B the change was -38.5 (95% CI=-78.9 to 1.94) g/day vs. 79.5 (95% CI=72.6-86.4) g/day (p<0.001). Similarly in Flock A the mean increase in CFLW between PD2 and PD3 was lower in ewe lambs with fetal loss than in those that maintained pregnancy (25.8 (95% CI=-13.8 to 65.4) g/day vs. 181.9 (95% CI=175.1-188.7) g/day) (p<0.001). There was no difference (p>0.05) in early and mid-gestation mean concentrations of progesterone in plasma between ewe lambs that had fetal loss compared with those that maintained pregnancy. CONCLUSIONS: Ewe lambs with fetal loss had a marked reduction in CFLW in the 24-30 days leading up to the identification of the fetal loss compared with those that maintained pregnancy. Early and mid-gestation concentrations of progesterone in plasma were similar between ewe lambs who went on to have fetal loss and those that maintained pregnancy.

The potential interaction between ewe body condition score and nutrition during very late pregnancy and lactation on the performance of twin-bearing ewes and their lambs.

OBJECTIVE: The present study aimed to determine the impact of ewe body condition score (BCS) (over a range of 2.0 to 3.0) and nutritional treatments (consisting of differing herbage masses) during very late pregnancy and lactation and their potential interaction on the performance of twin-bearing ewes and their lambs to weaning. METHODS: On day 142 of pregnancy, twin-bearing ewes with a BCS of 2.0, 2.5, or 3.0 were allocated to a "Moderate' or 'Unrestricted' nutritional treatment until day 95 of lactation (weaning). The nutritional treatments aimed to achieve average herbage masses of 1,200 to 1,300 kg dry matter (DM)/ha (Moderate) and 1,500 to 1,800 kg DM/ha (Unrestricted). RESULTS: There were no three-way interactions between ewe BCS group, nutritional treatment and time for any ewe or lamb parameter. The nutritional treatments had no effect (p>0.05) on lamb birth or weaning weight. Lambs born to Moderate ewes had greater survival and total litter weight at weaning (p<0.05). Regardless of BCS group, Unrestricted treatment ewes had greater body condition and back-fat depth at weaning than Moderate treatment ewes (p<0.05). Ewes of BCS 2.0 group reared lighter lambs to weaning (p<0.05) and tended to have a lower total litter weight (p = 0.06) than BCS 3.0 group ewes. CONCLUSION: This study suggests farmers should aim to have all ewes with a BCS of 2.5 or 3 in late pregnancy for optimal lamb weaning performance. Furthermore, there is no benefit to lamb production of offering ewes pasture masses >1,200 kg DM/ha during very late pregnancy and lactation.

Ad libitum Pasture Feeding in Late Pregnancy Does Not Improve the Performance of Twin-bearing Ewes and Their Lambs.

The present study evaluated the effect of controlled ryegrass-white clover herbage availability from day 128 until day 142 of pregnancy in comparison to unrestricted availability, on the performance of twin-bearing ewes of varying body condition score (BCS; 2.0, 2.5, or 3.0) and their lambs. It was hypothesised that under conditions of controlled herbage availability, the performance of lambs born to ewes with a greater BCS would be greater than those born to ewes with a lower BCS. During the period that the nutritional regimens were imposed, the pre- and post-grazing herbage masses of the Control regimen (1,070±69 and 801±30 kg dry matter [DM]/ha) were lower than the ad libitum regimen (1,784±69 and 1,333±33 kg DM/ha; p<0.05). The average herbage masses during lactation were 1,410±31 kg DM/ha. Nutritional regimen had no effect on ewe live weight, BCS and back fat depth or on lamb live weight, indices of colostrum uptake, maximal heat production, total litter weight weaned or survival to weaning (p>0.05). The difference in ewe BCSs and back fats observed among body condition groups was maintained throughout pregnancy (p<0.05). At weaning, ewes from the BCS2.0 group had lower BCS and live weight (2.4±0.2, 74.3±2.6 kg) than both the BCS2.5 (2.6±0.2, 78.6±2.4 kg) and BCS3.0 ewes (2.7±0.2, 79.0±2.6 kg; p<0.05), which did not differ (p>0.05). Ewe BCS group had no effect on lamb live weight at birth or weaning or on maximal heat production (p>0.05). Serum gamma glutamyl transferase concentrations of lambs born to BCS3.0 ewes were higher within 36 hours of birth than lambs born to BCS2.0 ewes and BCS2.5 ewes (51.8±1.9 vs 46.5±1.9 and 45.6±1.9 IU/mL, respectively [p<0.05]). There was, however, no effect of ewe body condition on lamb plasma glucose concentration (p>0.05). Lamb survival was the only lamb parameter that showed an interaction between ewe nutritional regimen and ewe BCS whereby survival of lambs born to BCS2.5 and BCS3.0 ewes differed but only within the Control nutritional regimen ewes (p<0.05). These results indicate farmers can provide twin-bearing ewes with pre- and post-grazing ryegrass-white clover herbage covers of approximately 1,100 and 800 kg DM/ha in late pregnancy, provided that herbage covers are 1400 in lactation, without affecting lamb performance to weaning. The present results also indicate that under these grazing conditions, there is little difference in ewe performance within the BCS range of 2.0 to 3.0 and therefore they do not need to be managed separately.

The selective prolyl hydroxylase inhibitor IOX5 stabilizes HIF-1α and compromises development and progression of acute myeloid leukemia.

Acute myeloid leukemia (AML) is a largely incurable disease, for which new treatments are urgently needed. While leukemogenesis occurs in the hypoxic bone marrow, the therapeutic tractability of the hypoxia-inducible factor (HIF) system remains undefined. Given that inactivation of HIF-1α/HIF-2α promotes AML, a possible clinical strategy is to target the HIF-prolyl hydroxylases (PHDs), which promote HIF-1α/HIF-2α degradation. Here, we reveal that genetic inactivation of Phd1/Phd2 hinders AML initiation and progression, without impacting normal hematopoiesis. We investigated clinically used PHD inhibitors and a new selective PHD inhibitor (IOX5), to stabilize HIF-α in AML cells. PHD inhibition compromises AML in a HIF-1α-dependent manner to disable pro-leukemogenic pathways, re-program metabolism and induce apoptosis, in part via upregulation of BNIP3. Notably, concurrent inhibition of BCL-2 by venetoclax potentiates the anti-leukemic effect of PHD inhibition. Thus, PHD inhibition, with consequent HIF-1α stabilization, is a promising nontoxic strategy for AML, including in combination with venetoclax.

Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5.

Jumonji-C (JmjC) domain-containing protein 5 (JMJD5) is a human 2-oxoglutarate (2OG) and Fe(ii)-dependent oxygenase which catalyses the post-translational C3 hydroxylation of arginyl-residues and which is linked to the circadian rhythm and to cancer biology through as yet unidentified mechanisms. We report robust solid phase extraction coupled to mass spectrometry (SPE-MS)-based JMJD5 assays which enable kinetic and high-throughput inhibition studies. The kinetic studies reveal that some synthetic 2OG derivatives, notably including a 2OG derivative with a cyclic carbon backbone (i.e. (1R)-3-(carboxycarbonyl)cyclopentane-1-carboxylic acid), are efficient alternative cosubstrates of JMJD5 and of factor inhibiting hypoxia-inducible transcription factor HIF-α (FIH), but not of the Jumonji-C (JmjC) histone Nε-methyl lysine demethylase KDM4E, apparently reflecting the closer structural similarity of JMJD5 and FIH. The JMJD5 inhibition assays were validated by investigating the effect of reported 2OG oxygenase inhibitors on JMJD5 catalysis; the results reveal that broad-spectrum 2OG oxygenase inhibitors are also efficient JMJD5 inhibitors (e.g. N-oxalylglycine, pyridine-2,4-dicarboxylic acid, ebselen) whereas most 2OG oxygenase inhibitors that are in clinical use (e.g. roxadustat) do not inhibit JMJD5. The SPE-MS assays will help enable the development of efficient and selective JMJD5 inhibitors for investigating the biochemical functions of JMJD5 in cellular studies.

The Behaviour of Sheep around a Natural Waterway and Impact on Water Quality during Winter in New Zealand.

Access of livestock, such as cattle, to waterways has been shown to be a cause of poor water quality due to pugging damage and excretion entering the water. In New Zealand, regulations require that cattle, deer, and pigs are excluded from accessing waterways, but there are no such requirements for sheep. The current study utilised 24 h video cameras, global positioning system units, and triaxial accelerometers to observe the interaction of Romney ewes (n = 40) with a natural waterway. Ewes were either restricted (week 1) or given access to a reticulated water trough (week 2). Proximity data showed that ewes spent more time within 3 m of the waterway when the trough was unrestricted than when restricted (14.1 ± 5.7 and 10.8 ± 5.1 min/ewe/day, respectively; p < 0.05). Ewes travelled shorter distances on the steeper areas of paddock than flatter areas. Similarly, ewes showed a spatial preference for the flat and low sloped areas of the paddock. Concentrations of suspended sediment and total phosphorus were higher during access to a reticulated water trough which coincided with the week with more rainy days. Phosphorus and E. coli concentrations in the stream water samples were the above recommended Australian and New Zealand Environment and Conservation Council water quality guidelines, especially after rainy days, but did not appear to be directly related to sheep activity. Overall, the results suggest that during winter, ewes interacted very little with the waterway and were thus unlikely to influence the levels of nutrient and pathogens in the waterway.

5-Substituted Pyridine-2,4-dicarboxylate Derivatives Have Potential for Selective Inhibition of Human Jumonji-C Domain-Containing Protein 5.

Jumonji-C domain-containing protein 5 (JMJD5) is a 2-oxoglutarate (2OG)-dependent oxygenase that plays important roles in development, circadian rhythm, and cancer through unclear mechanisms. JMJD5 has been reported to have activity as a histone protease, as an Nε-methyl lysine demethylase, and as an arginine residue hydroxylase. Small-molecule JMJD5-selective inhibitors will be useful for investigating its (patho)physiological roles. Following the observation that the broad-spectrum 2OG oxygenase inhibitor pyridine-2,4-dicarboxylic acid (2,4-PDCA) is a 2OG-competing JMJD5 inhibitor, we report that 5-aminoalkyl-substituted 2,4-PDCA derivatives are potent JMJD5 inhibitors manifesting selectivity for JMJD5 over other human 2OG oxygenases. Crystallographic analyses with five inhibitors imply induced fit binding and reveal that the 2,4-PDCA C5 substituent orients into the JMJD5 substrate-binding pocket. Cellular studies indicate that the lead compounds display similar phenotypes as reported for clinically observed JMJD5 variants, which have a reduced catalytic activity compared to wild-type JMJD5.