Bridging the age gap in breast cancer: cluster randomized trial of two decision support interventions for older women with operable breast cancer on quality of life, survival, decision quality, and treatment choices.

BACKGROUND: Rates of surgery and adjuvant therapy for breast cancer vary widely between breast units. This may contribute to differences in survival. This cluster RCT evaluated the impact of decision support interventions (DESIs) for older women with breast cancer, to ascertain whether DESIs influenced quality of life, survival, decision quality, and treatment choice. METHODS: A multicentre cluster RCT compared the use of two DESIs against usual care in treatment decision-making in older women (aged at least ≥70 years) with breast cancer. Each DESI comprised an online algorithm, booklet, and brief decision aid to inform choices between surgery plus adjuvant endocrine therapy versus primary endocrine therapy, and adjuvant chemotherapy versus no chemotherapy. The primary outcome was quality of life. Secondary outcomes included decision quality measures, survival, and treatment choice. RESULTS: A total of 46 breast units were randomized (21 intervention, 25 usual care), recruiting 1339 women (670 intervention, 669 usual care). There was no significant difference in global quality of life at 6 months after the baseline assessment on intention-to-treat analysis (difference -0.20, 95 per cent confidence interval (C.I.) -2.69 to 2.29; P = 0.900). In women offered a choice of primary endocrine therapy versus surgery plus endocrine therapy, knowledge about treatments was greater in the intervention arm (94 versus 74 per cent; P = 0.003). Treatment choice was altered, with a primary endocrine therapy rate among women with oestrogen receptor-positive disease of 21.0 per cent in the intervention versus 15.4 per cent in usual-care sites (difference 5.5 (95 per cent C.I. 1.1 to 10.0) per cent; P = 0.029). The chemotherapy rate was 10.3 per cent at intervention versus 14.8 per cent at usual-care sites (difference -4.5 (C.I. -8.0 to 0) per cent; P = 0.013). Survival was similar in both arms. CONCLUSION: The use of DESIs in older women increases knowledge of breast cancer treatment options, facilitates shared decision-making, and alters treatment selection. Trial registration numbers: EudraCT 2015-004220-61 (https://eudract.ema.europa.eu/), ISRCTN46099296 (http://www.controlled-trials.com).

Animal grazing selectivity and plant chemistry issues impact on the potential of Rhagodia preissii as an anthelmintic shrub.

Rhagodia preissii had shown significant in vitro anthelmintic activity in a previous study, we examined the effect of including this shrub in the diet of sheep infected with Trichostrongylus colubriformis. Worm-infected merino wethers were grazed for 7 weeks on either R. preissii or annual pasture, and faecal egg counts (FECs) were conducted weekly. Plant material was collected weekly from eaten and uneaten plants, and analysed for levels of plant secondary metabolites (tannins, oxalates, saponins) and in vitro anthelmintic activity. While mean FECs were consistently lower in sheep grazing R. preissii compared to pasture (reductions of 20-74%), the differences were not significant. There was no relationship between grazing preference (eaten or uneaten) and in vitro anthelmintic activity of plant extracts. The levels of saponins and oxalates did not correlate with grazing preference or in vitro anthelmintic activity, while tannins were not responsible for the anthelmintic activity. While the identity of the grazing deterrent and in vitro anthelmintic compounds remain unknown, the presence of plants which were both highly preferred by the sheep and showed in vitro anthelmintic activity indicates a potential to develop the species as an anthelmintic shrub through selection of shrub populations dominated by such plants.

Exploring the anthelmintic properties of Australian native shrubs with respect to their potential role in livestock grazing systems.

We measured in vitro anthelmintic activity in extracts from 85 species of Australian native shrub, with a view to identifying species able to provide a degree of worm control in grazing systems. Approximately 40% of the species showed significant activity in inhibiting development of Haemonchus contortus larvae. The most active extracts showed IC50 values of 60-300 microg/ml. Pre-incubation with polyvinylpolypyrrolidine removed the activity from some extracts, implicating tannins as the bioactive agent, while in other cases the pre-incubation had no effect, indicating the presence of other anthelmintic compounds. Plant reproductive maturity (onset of flowering or fruiting) was associated with increasing anthelmintic activity in some species. Variability was observed between plants of the same species growing in different environments, while variation between individual plants of the same species within a single field suggests the existence of distinct chemotypes. Significant activity against adult H. contortus worms in vitro was also demonstrated in a limited number of extracts tested against this life stage. Our study indicates that there is potential for Australian native shrubs to play an anthelmintic role in grazing systems, and highlights some plant biology factors which will need to be considered in order to maximize any anthelmintic effects.

Iron content of eggs from hens given diets containing organic forms of iron, serine and methyl group donors, or phytoestogens.

1. This investigation included three experiments to determine whether the iron content of egg yolks could be enriched by supplementation of the laying diet with iron bound to organic compounds (Experiment 1), serine and methyl group donors methionine and choline (Experiment 2) or phytoestrogens (Experiment 3). 2. Hens at 34, 54 and 56 weeks of age were given experimental diets for 6, 4 and 4 weeks, respectively, in Experiments 1-3. Yolks from eggs laid over three successive days in the final week of feeding were pooled for each hen for analysis of iron by inductively coupled plasma mass spectrometry. 3. Iron concentration in egg yolk averaged 68-70, 66-71 and 62-69 microg/g in the respective experiments. 4. The addition of bloodmeal (1.22 mg Fe/egg) or phytoestrogens (1.25 mg Fe/egg) increased the total iron content of yolks by over 15% compared with the control diet (1.10 mg Fe/egg), and although this increase was not statistically significant it suggests that the iron content of eggs could be sufficiently manipulated to justify a nutritional claim of iron enrichment.

Increasing the age of gilts at first boar contact improves the timing and synchrony of the pubertal response but does not affect potential litter size.

One hundred and ninety-two Large White/Landrace crossbred gilts were used in this study. The study was conducted in two blocks, with 16 gilts allocated to each of six treatments in each block. The treatments compared the effects on puberty attainment of commencing boar exposure at 161, 182 or 203 days of age, and the effect of first mating gilts at either the pubertal or second oestrus on ovulation rate and early embryo survival. Boar contact took place in a detection-mating area (DMA), and consisted of 20 min/day of full contact with a vasectomized boar greater than ten months of age. Gilts were artificially inseminated at the allocated oestrus, with the reproductive tracts collected at 22.8+/-0.4 days after first mating (mean+/-S.E.M.), and the numbers of corpora lutea and viable embryos recorded. The age at which gilts attained puberty increased with the age at which boar exposure commenced. Mean gilt ages at puberty were 179.5+/-1.6, 191.7+/-1.2 and 210.3+/-0.9 days, respectively, when boar contact commenced at 161, 182 and 203 days of age, P<0.01. Mean days-to-puberty was significantly shorter when boar contact began at 182 and 203 days of age compared to 161 days of age (10.4+/-1.2 and 8.3+/-0.9 days versus 18.9+/-1.5 days, respectively, P<0.01). Similarly, commencing boar exposure at 182 or 203 days of age as opposed to 161 days of age significantly increased the proportion of gilts attaining puberty within 10 days of start of boar exposure (0.67 and 0.70 versus 0.24, P<0.01). Mean days-to-puberty and the synchrony of puberty attainment were similar when boar contact commenced at 182 and 203 days of age. There was no significant effect of mating oestrus or age of gilts at mating on ovulation rate, embryo number or embryo survival. In conclusion, the current data indicate that the timing and synchrony of puberty attainment is significantly improved when first boar exposure of gilts is delayed until 182 days of age. Further, it is evident that within the age range investigated, delaying first mating until the second oestrus does not significantly increase either ovulation rate or embryo number at day 20 post-mating.

Sheep deficient in vitamin E preferentially select for a feed with a higher concentration of vitamin E.

Given the capacity of ruminants to modify diet selection based on metabolic needs, we hypothesised that, when given a choice, lambs experiencing a vitamin E deficiency would consume more of a vitamin E-enriched feed than lambs not deficient in vitamin E. Fifty-six Dohne Merino lambs were divided into two groups and fed either a vitamin E-deficient diet over 40 days to induce low plasma vitamin E or a vitamin E-enriched diet to induce high plasma vitamin E. The lambs were then offered a choice of vitamin E-enriched and vitamin E-deficient pellets. For half of the animals, the enriched diet was paired with strawberry flavour and the deficient diet was paired with orange flavour, while the reverse pairings were offered to the others. Lamb preference for the diets was measured daily for the following 15 days. There was a three-way interaction between the high and low vitamin E treatment groups×vitamin E content and type of flavour in the feed×time (days). The lambs preferred pellets flavoured with strawberry but this preference changed to orange flavour in vitamin E-deficient lambs if the orange flavour was paired with high vitamin E. Lambs without a deficiency continued to prefer strawberry-flavoured pellets, regardless of the vitamin E concentrations in the pellets. It is possible that self-learning contributed to the low vitamin E group of lambs changing preference to orange flavour in order to consume more vitamin E, presumably to remediate the deficiency.

Nucleotide sequence and expression in Escherichia coli of cDNAs encoding papaya proteinase omega from Carica papaya.

We have cloned and sequenced two similar, but distinct, cDNAs from both fruit and leaf tissues of Carica papaya. The C-terminal portion of the predicted amino acid (aa) sequence of one of the clones has complete identity with the mature enzyme sequence of the cysteine proteinase papaya proteinase omega (Pp omega). The second clone contains ten individual bp changes compared with the first and encodes a protein with three single-aa substitutions, only one of which is located in the mature sequence, but most noticeably carries an additional 19-aa C-terminal extension. The clones encode pre-pro precursor isoforms of Pp omega. The former of these clones has been expressed in Escherichia coli using a T7 polymerase expression system to produce insoluble pro-enzyme which has been solubilized and refolded to yield auto-activable pro-Pp omega.

Functional characterization of agonists at recombinant human 5-HT2A, 5-HT2B and 5-HT2C receptors in CHO-K1 cells.

1. The goal of this study was to characterize the agonist pharmacology of human 5-HT2A, 5-HT2B and 5-HT2C (VSV) receptors expressed in CHO-K1 (Chinese hamster ovary) cells. 2. We used a fluorometric imaging plate reader (FLIPR) which allows rapid detection of rises in intracellular calcium levels upon the addition of agonists. 3. Stimulation of all three receptors by 5-HT caused a robust concentration dependent increase in intracellular calcium levels. No such effect was observed from non-transfected control CHO-K1 cells. 4. The rank order of potency of agonists at the different receptor subtypes varied. Tryptamines, BW-723C86, d-norfenfluramine, Ro 60-0175 and LSD exhibited the following rank order of potency; 5-HT2B>5-HT2C>5-HT2A. Piperazines such as m-Chlorophenylpiperazine (mCPP), ORG-12962, MK-212 and also ORG-37684 exhibited a rank order of potency of 5-HT2C>5-HT2B>5-HT2A. The phenylisopropylamines DOI and DOB had a rank order of 5-HT2A>5-HT2B>5-HT2C. 5. Many agonists tested had partial agonist actions when compared to 5-HT, and a wide range of relative efficacies were exhibited, which was cell line dependent. For example, mCPP had a relative efficacy of 65% at 5-HT2C receptors but <25% at either 5-HT2A or 5-HT2B receptors. 6. Interpretation of literature values of functional assays using different cell lines, different receptor expression levels and different receptor isoforms, is complex. Species differences and the previous use of antagonist radioligands to characterize agonist potency in binding assays emphasizes the importance of studying agonists in the same experiment using the same assay conditions and parental cell lines.

Agonist-induced functional desensitization of recombinant human 5-HT2 receptors expressed in CHO-K1 cells.

The desensitization characteristics of recombinant human 5-HT(2A), 5-HT(2B), and 5-HT(2C) receptors (VSV and INI isoforms) stably expressed in CHO-K1 (Chinese hamster ovary) cells was investigated by calcium fluorimetry. Comparative desensitization characteristics of the agonists 5-HT, m-chlorophenylpiperazine (mCPP), and 2,5-dimethoxy-4-iodoamphetamine hydrobromide (DOI) were performed. Human 5-HT(2C (INI)) receptors exhibited a greater degree of desensitization to all agonists tested than edited 5-HT(2C (VSV)) receptors. A 2-hr exposure to 5-HT resulted in a significantly larger reduction in response upon re-exposure to 5-HT at 5-HT(2C (INI)) receptors, as compared to 5-HT(2C (VSV)) receptors (72% and 47% respectively, P < 0.01). Both receptor isoforms were expressed at similar densities. Human 5-HT(2B) receptors exhibited the most dramatic degree of desensitization, with prior exposure to 5-HT reducing subsequent response to 5-HT by 80%, with an extremely rapid time-course (t(1/2) < 5 min). The response at 5-HT(2A) receptors was reduced by 54%. The partial agonists mCPP and DOI also elicited desensitization, generally in line with their relative efficacies at each receptor, but exhibited more rapid kinetic profiles than 5-HT. Heterologous desensitization of an endogenously expressed G(q/11)-coupled purinergic receptor was also examined following preincubation of the cell lines with 10 microM 5-HT. Only stimulation of 5-HT(2C (VSV)) receptors resulted in a profound attenuation of subsequent ATP mediated responses. These results demonstrate differing degrees of both homologous and heterologous desensitization of 5-HT(2) receptors. Additionally, the different desensitization profiles of 5-HT(2C (INI)) and 5-HT(2C (VSV)) receptor may be due to signal transduction differences caused by RNA editing.

Characterisation of agonist binding on human 5-HT2C receptor isoforms.

The 5-HT2C receptor is expressed in different isoforms as a result of mRNA editing. Both INI (unedited) and VSV (a fully edited version) isoforms are abundant in rat brain. The VSV isoform lacks the high affinity recognition site for 5-HT, which may be caused by low efficiency coupling to G-proteins. In this study we have investigated the pharmacology of the agonist binding site of these two isoforms of the 5-HT2C receptor. The VSV isoform was expressed in Chinese hamster ovary cells (CHO) and the INI isoform in both Chinese hamster ovary cells and human embryonic kidney cells (HEK-293). Saturation analysis using [3H]5-HT revealed high and low affinity recognition sites on the INI isoform in both cell types whilst the VSV isoform did not have the high affinity binding site for [3H]5-HT. Displacement studies were undertaken using [3H]5-HT to label the receptors. In these studies the affinity of agonists (5-HT, Ro600175 ((S)-2-(6-Chloro-5-fluoroindol-1-yl)-1-methylethylamine), MK212 (6-Chloro-2-(piperazinyl) pyrazine), mCPP (1-(m-chlorophenyl)-piperazine), TfMPP (N-(m-trifluoromethylphenyl)piperazine), DOI (1-(2,5-Dimethoxy-4-iodophenyl)-2-aminopropane), DOB (1-(4-bromo-2,5-dimethoxyphenyl)-2-aminopropane) and 8OH-DPAT (8-hydroxy-2-(di-N-propylamino)tetralin) was higher at the INI isoform, whilst antagonist affinity (ketanserin and mesulergine) did not change between the two receptor isoforms. There were no differences between the INI isoform expressed in the CHO and HEK-293. This suggests that the INI isoform of the 5-HT2C receptor is pharmacologically similar to the VSV form of the 5-HT2C receptor but that it couples more efficiently to G-proteins.

Metabolic responses to mid-pregnancy shearing that are associated with a selective increase in the birth weight of twin lambs.

Previous studies have shown that shearing pregnant ewes at mid- or late-pregnancy is associated with an increase in lamb birth weight. The present study was designed to investigate metabolic responses that may underlie this response. Single- and twin-bearing ewes were either unshorn or shorn at mid-pregnancy (Day 69 of pregnancy; P69), and insulin, glucose and epinephrine challenges were conducted on P109-111 and P132-134. Shearing increased the birth weight of twin lambs by over 1 kg (P < 0.001) without having any effect on singleton birth weight. This response was associated with a 10-20% reduction in the insulin response to a glucose challenge (P < 0.05) without a change in glucose clearance following either glucose or insulin challenges. The lipolytic response to epinephrine challenge increased as pregnancy progressed, but was not associated with the increased birth weight of twin lambs born to shorn ewes. By late pregnancy, a 25% reduction in maternal IGF-I concentration and a two- to threefold increase in maternal IGFBP-1 concentration (P < 0. 05) associated with shearing were observed. The increase in lamb birth weight associated with mid-pregnancy shearing may have been associated with an increase in the non-insulin dependent uptake of glucose by the placental-fetal unit.

Altered secretion of LH does not explain seasonal effects on early pregnancy in gilts.

This work was undertaken to study the effects of energy intake and gut fill on LH secretion in the early pregnant gilt to investigate the role of the hypothalamo-pituitary-gonadal axis in the seasonal disruption of pregnancy. Four groups of mated gilts (N = 23 in total) were individually housed and fed four different energy levels (23, 46, 47 and 55 megajoules digestible energy per day, MJ DE day-1) for two weeks after mating during the season identified as a period of increased rate of early pregnancy failures from January till April (summer-autumn). The energy content of base feed (13 MJ kg-1) was increased by means of adding a fat supplement (soybean oil) to the feed of two groups of gilts, while the other two groups were given different levels of the basic feed. The groups gained weight and backfat according to energy intake levels during the two-week period. Frequent blood samples (15-min interval) collected on day 14 revealed a significant treatment effect on LH pulse amplitude: the group on the highest energy intake level (55 MJ DE day-1) had higher LH pulse amplitude compared with the other groups (1.01 +/- 0.04, 0.74 +/- 0.04, 0.72 +/- 0.08 and 0.66 +/- 0.05 ng ml-1 in the descending order of energy intake, P < 0.01). There was no effect of energy intake on LH pulse frequency, mean level, area under the curve or mean nadir (P > 0.10). Plasma insulin concentrations tended to increase (P < 0.08) in each group with time after feeding, but no significant differences between the treatment groups were found. Plasma free fatty acid (FFA) concentrations were decreased 1 h after feeding (P < 0.05) in all but the group on 55 MJ DE day-1. This group also had higher postprandial FFA concentrations in comparison with other groups (P < 0.05). In conclusion, these results show that energy density of feed and energy intake have little effect on LH secretion in the early pregnant gilt. Protective effect of high feeding level against seasonal disruption of pregnancy appears to be mediated by mechanisms other than an alteration in LH secretion.

Body composition at farrowing and nutrition during lactation affect the performance of primiparous sows: I. Voluntary feed intake, weight loss, and plasma metabolites.

We used 35 primiparous sows to investigate the link between body fatness at farrowing and voluntary feed intake (VFI) during lactation. Two groups of sows were fed differently throughout gestation (either 2.3 kg/d of a diet containing 5.8% CP and 14.6 MJ DE/kg as fed or 1.7 kg/d of a diet containing 15.6% CP and 14.5 MJ DE/kg as fed) so that they commenced lactation at a similar body weight (158 to 152 kg) but with different body compositions: either 340 (fat) or 280 (lean) g of body fat/kg BW (P < .001). During lactation, sows were offered either a low-protein diet (7.9% CP and 15.5 MJ DE/kg as fed) or a high-protein diet (19.0% CP and 15.6 MJ DE/kg as fed) on an ad libitum basis. During lactation, VFI was measured daily, and sow body weight and backfat were measured weekly. Blood samples were collected from sows on d 110 of gestation and d 14 and 28 of lactation, and plasma was analyzed for NEFA, glycerol, insulin, glucose, and beta-hydroxybutyrate. Fat sows ate 30% less than their lean counterparts during lactation (P < .001), which corresponded to a 70% higher concentration of NEFA in plasma (P = .01) and a 30% higher concentration of glycerol (P = .15). The VFI during the first 2 wk of lactation was affected only by body fatness and not by the protein content of the lactation diet. The dietary supply of protein influenced VFI during wk 3 and 4 of lactation, possibly by affecting milk production and hence the drive to consume feed. Weight loss, particularly lean tissue loss, was minimized by feeding the high-protein diet during lactation (P < .002).

Body composition at farrowing and nutrition during lactation affect the performance of primiparous sows: II. Milk composition, milk yield, and pig growth.

Sows that were either fat or lean at farrowing (340 or 280 g of body fat/kg BW, respectively) were offered either a low-protein (LP; 7.9% CP and 15.5 MJ DE/kg as fed) or a high-protein (HP; 19.0% CP and 15.6 MJ DE/kg as fed) diet on an ad libitum basis throughout a 4-wk lactation to test the hypothesis that the amount of milk and its composition are responsive to the supply of endogenous (body reserves) and exogenous (diet) substrates. Pigs were weighed at birth and weekly during lactation, milk yield was estimated using deuterium oxide in early (d 4 to 8) and late lactation (d 24 to 28), and milk samples were collected to determine composition in early (d 4 to 6) and late lactation (d 25 to 27). Throughout lactation, milk yield and composition were mainly associated with differences in litter size. Milk yield was about 15% higher in lean than in fat sows and in sows fed HP rather than LP, but large CV (17 to 32%) prevented these differences from attaining significance (P > .273). The responses in milk yield were reflected in pig growth. Differences in milk composition between treatment groups were not significant; however, during early lactation there was a tendency for fat sows to produce milk with a fat content 21% higher, and a protein content 12% lower, than that of lean sows. Changes in the protein:energy ratio of milk during the course of lactation and small changes in milk yield composition collectively suggested that in early lactation, sow body composition affected milk production but, as lactation progressed, the dietary supply of precursors for milk synthesis became more important.

Feed intake of sheep when allowed ad libitum access to feed in methane respiration chambers.

The patterns of feed intake when animals are allowed ad libitum access to feed in a respiration chamber is not known, nor are the potential effects of the artificial environment of chambers on voluntary feed intake. The objectives of the study were to describe the pattern of hourly feed intake of sheep when fed for ad libitum intake in respiration chambers and determine the repeatability of this pattern and the correlation between feed intake and methane production calculated at hourly intervals. Daily and hourly measurements of methane production and feed intake of 47 Merino wethers were measured in respiration chambers twice, 4 wk apart. We found that hourly feed intake of sheep with ad libitum access to feed in respiration chambers showed a repeatable pattern over the 2 measurement periods (r = 0.89, P < 0.001). During both measurements, sheep ate continuously throughout the 23 h period, but most of the eating occurred during the first 8 h in the respiration chambers. There was a significant linear correlation (r = 0.22) between hourly feed intake and hourly methane production (P < 0.001). An unexpected result from this study was that despite using an accepted and published acclimatization procedure to habituate the animals to the respiration chambers, sheep had 15 to 25% lower feed intake in the respiration chambers compared with their feed intake during the previous week in the animal house pens. In addition, daily feed intake in the respiration chamber was not correlated with feed intake in any of the 7 d before entering the chamber (P > 0.05). Future methane research may consider using feed intake and changes in intake levels as a quantitative indicator of habituation to the methane measurement procedure and environment, which, given the tight association between feed intake and methane production, will be crucial in providing accurate values for methane production.

Consumption of a high-salt diet by ewes during pregnancy alters nephrogenesis in 5-month-old offspring.

Maternal nutrition during pregnancy can affect kidney development in the foetus, which may lead to adverse consequences in the mature kidney. It was expected that high-salt intake by pregnant ewes would lead to a reduction in foetal glomerular number but that the ovine kidney would adapt to maintain homoeostasis, in part by increasing the size of each glomerulus. Merino ewes that were fed either a control (1.5% NaCl) or high-salt (10.5% NaCl) diet during pregnancy, as well as their 5-month-old offspring, were subjected to a dietary salt challenge, and glomerular number and size and sodium excretion were measured. The high-salt offspring had 20% fewer glomeruli compared with the control offspring (P < 0.001), but they also had larger glomerular radii compared with the control offspring (P < 0.001). Consequently, the cross-sectional area of glomeruli was 18% larger in the high-salt offspring than in the control offspring (P < 0.05). There was no difference in the daily urinary sodium excretion between the two offspring groups (P > 0.05), although the high-salt offspring produced urine with a higher concentration of sodium. Our results demonstrated that maternal high-salt intake during pregnancy affected foetal nephrogenesis, altering glomerular number at birth. However, the ability to concentrate and excrete salt was not compromised, which indicates that the kidney was able to adapt to the reduction in the number of glomeruli.

Nutritional restriction of pre-pubertal liveweight gain impairs ovarian follicle growth and oocyte developmental competence of replacement gilts.

The objective was to investigate the effects of moderate restriction of pre- and peri-pubertal liveweight gain on ovarian development and oocyte meiotic competence. At 70 d of age, and 27.7 ± 0.4 kg liveweight (LW), 64 Large White/Landrace crossbred gilts were allocated to two treatment groups (n = 32 gilts/treatment); one group was fed to attain a LW of 70 kg at 161 d of age (LIGHT), while the other group was fed to reach 100 kg LW (HEAVY). At 161 d of age, half of the gilts in each group (n = 16) were fed to gain LW at 0.5 kg/d (LOW), while the remaining half (n = 16) were fed to gain LW at 1.0 kg/d (HIGH) between 161 and 175 d of age, at which point they were killed and ovaries collected. For each gilt, surface antral follicles were counted and aspirated according to three size categories: 1-2.9 mm (small); 3-6 mm (medium); and > 6 mm (large). Follicles were pooled for each size class and treatment. Cumulus-oocyte-complexes (COC) recovered from small and medium follicles were matured in vitro (IVM) for 44 to 46 h, and meiotic maturation assessed. There was an effect of treatment (LIGHT versus HEAVY) on the number of medium sized follicles: 25.1 ± 2.59 versus 34.3 ± 2.60 (P < 0.05). The ovaries of LOW gilts had more small follicles and fewer medium follicles compared to those of HIGH gilts: 92.8 ± 8.35 versus 59.8 ± 5.24, and 25.1 ± 2.59 versus 32.5 ± 2.86 (P < 0.05). Target LW at 161 d did not affect meiotic progression of oocytes. However, LOW compared to HIGH LW gain between 161 and 175 d resulted in fewer oocytes reaching MII (0.40 versus 0.54; P < 0.05). In conclusion, moderately restricting feed intake impaired follicle growth beyond 3 mm and reduced oocyte meiotic competence. Further, although a carry-over effect of long-term feed restriction on follicle growth was evident, acute changes in feed intake during the 14 d prior to ovary collection had the greatest effect on oocyte nuclear maturation in vitro.

Offspring born to ewes fed high salt during pregnancy have altered responses to oral salt loads.

Prenatal growth is sensitive to the direct and indirect effects of maternal dietary intake; manipulation can lead to behavioural and physiological changes of the offspring later in life. Here, we report on three aspects of how a high-salt diet during pregnancy (conception to parturition) may affect the offspring's response to high oral salt loads: (i) dietary preferences for salt; (ii) response to salt and water balance and aldosterone and arginine vasopressin (AVP) concentrations after an oral salt challenge; (iii) concentrations of insulin and leptin after an oral salt challenge. We used two groups of lambs born to ewes fed either a high-salt (13% NaCl) diet during pregnancy (S lambs; n = 12) or control animals born to ewes fed a conventional (0.5% NaCl) diet during pregnancy (C lambs; n = 12). Lambs were subjected to short- (5 min) and long-term (24 h) preference tests for a high-salt (13% NaCl) or control diet, and the response to an oral challenge with either water or 25% NaCl solution were also carried out. Weaned lambs born to ewes fed high salt during pregnancy did not differ in their preference for dietary salt, but they did differ in their physiological responses to an oral salt challenge. Results indicate that these differences reflect an alteration in the regulation of water and salt balance as the metabolic hormones, insulin and leptin, were not affected. During the first 2 h after a single salt dose, S lambs had a 25% lower water intake compared to the C lambs. S lambs had, on average, a 13% lower AVP concentration than the C lambs (P = 0.014). The plasma concentration of aldosterone was higher in the S lambs than in the C lambs (P = 0.013). Results suggest that lambs born to ewes that ingest high amounts of salt during pregnancy are programmed to have an altered thirst threshold, and blunted response in aldosterone to oral salt loads.