Child body mass index, genotype and parenting in the prediction of restrictive feeding.

BACKGROUND: Restrictive feeding is implicated in pediatric obesity, and caregivers increase controlling feeding practices on the basis of higher child weight status. However, few studies have examined how child genetic and parenting characteristics together impact restrictive feeding. OBJECTIVES: We examined whether child body mass index (BMI) status predicts caregiver use of restrictive feeding and if this association is moderated by (i) caregiver strategies to manage their children's distress and (ii) child variations in the catechol-O-methyltransferase (COMT) gene (Val158 Met, rs4680). METHODS: Participants included 126 Caucasian children (50% girls) and their caregivers who were participating in a larger study in the USA. Caregivers reported on their feeding practices and responses to child distress when children were 2.5-3.5 years of age. Child anthropometric measurements were also obtained. Restrictive feeding was assessed again 1-1.5 years later. Genomic DNA was obtained from saliva samples, and COMT-rs4680 was genotyped using TaqMan® methodology. RESULTS: Child BMI percentile predicted subsequent caregiver restrictive feeding for children who were Met/Met and who had caregivers reporting higher use of negative responses to child distress. For Val carriers, BMI percentile predicted restrictive feeding when caregivers were below the mean on these responses. CONCLUSIONS: Caregivers are at risk for use of restrictive feeding practices when their children are at higher BMI percentiles, and this association increases when caregivers use more ineffective stress regulation practices and their children are homozygous for the Met allele. Prevention programmes might focus on parenting behaviours that foster emotion regulation and consider variation in child responses to parenting.

Electroconvulsive therapy modulates plasma pigment epithelium-derived factor in depression: a proteomics study.

Electroconvulsive therapy (ECT) is the most effective treatment for severe depression, yet its mechanism of action is not fully understood. Peripheral blood proteomic analyses may offer insights into the molecular mechanisms of ECT. Patients with a major depressive episode were recruited as part of the EFFECT-Dep trial (enhancing the effectiveness of electroconvulsive therapy in severe depression; ISRCTN23577151) along with healthy controls. As a discovery-phase study, patient plasma pre-/post-ECT (n=30) was analyzed using 2-dimensional difference in gel electrophoresis and mass spectrometry. Identified proteins were selected for confirmation studies using immunodetection methods. Samples from a separate group of patients (pre-/post-ECT; n=57) and matched healthy controls (n=43) were then used to validate confirmed changes. Target protein mRNA levels were also assessed in rat brain and blood following electroconvulsive stimulation (ECS), the animal model of ECT. We found that ECT significantly altered 121 protein spots with 36 proteins identified by mass spectrometry. Confirmation studies identified a post-ECT increase (P<0.01) in the antiangiogenic and neuroprotective mediator pigment epithelium-derived factor (PEDF). Validation work showed an increase (P<0.001) in plasma PEDF in depressed patients compared with the controls that was further increased post-ECT (P=0.03). PEDF levels were not associated with mood scores. Chronic, but not acute, ECS increased PEDF mRNA in rat hippocampus (P=0.02) and dentate gyrus (P=0.03). This study identified alterations in blood levels of PEDF in depressed patients and further alterations following ECT, as well as in an animal model of ECT. These findings implicate PEDF in the biological response to ECT for depression.

Cell-specific abnormalities of glutamate transporters in schizophrenia: sick astrocytes and compensating relay neurons?

Excitatory amino-acid transporters (EAATs) bind and transport glutamate, limiting spillover from synapses due to their dense perisynaptic expression primarily on astroglia. Converging evidence suggests that abnormalities in the astroglial glutamate transporter localization and function may underlie a disease mechanism with pathological glutamate spillover as well as alterations in the kinetics of perisynaptic glutamate buffering and uptake contributing to dysfunction of thalamo-cortical circuits in schizophrenia. We explored this hypothesis by performing cell- and region-level studies of EAAT1 and EAAT2 expression in the mediodorsal nucleus of the thalamus in an elderly cohort of subjects with schizophrenia. We found decreased protein expression for the typically astroglial-localized glutamate transporters in the mediodorsal and ventral tier nuclei. We next used laser-capture microdissection and quantitative polymerase chain reaction to assess cell-level expression of the transporters and their splice variants. In the mediodorsal nucleus, we found lower expression of transporter transcripts in a population of cells enriched for astrocytes, and higher expression of transporter transcripts in a population of cells enriched for relay neurons. We confirmed expression of transporter protein in neurons in schizophrenia using dual-label immunofluorescence. Finally, the pattern of transporter mRNA and protein expression in rodents treated for 9 months with antipsychotic medication suggests that our findings are not due to the effects of antipsychotic treatment. We found a compensatory increase in transporter expression in neurons that might be secondary to a loss of transporter expression in astrocytes. These changes suggest a profound abnormality in astrocyte functions that support, nourish and maintain neuronal fidelity and synaptic activity.

Glutamate transporter splice variant expression in an enriched pyramidal cell population in schizophrenia.

Dysregulation of the glutamate transporters EAAT1 and EAAT2 and their isoforms have been implicated in schizophrenia. EAAT1 and EAAT2 expression has been studied in different brain regions but the prevalence of astrocytic glutamate transporter expression masks the more subtle changes in excitatory amino acid transporters (EAATs) isoforms in neurons in the cortex. Using laser capture microdissection, pyramidal neurons were cut from the anterior cingulate cortex of postmortem schizophrenia (n = 20) and control (n = 20) subjects. The messenger RNA (mRNA) levels of EAAT1, EAAT2 and the splice variants EAAT1 exon9skipping, EAAT2 exon9skipping and EAAT2b were analyzed by real time PCR (RT-PCR) in an enriched population of neurons. Region-level expression of these transcripts was measured in postmortem schizophrenia (n = 25) and controls (n = 25). The relationship between selected EAAT polymorphisms and EAAT splice variant expression was also explored. Anterior cingulate cortex pyramidal cell expression of EAAT2b mRNA was increased (P < 0.001; 67%) in schizophrenia subjects compared with controls. There was no significant change in other EAAT variants. EAAT2 exon9skipping mRNA was increased (P < 0.05; 38%) at region level in the anterior cingulate cortex with no significant change in other EAAT variants at region level. EAAT2 single-nucleotide polymorphisms were significantly associated with changes in EAAT2 isoform expression. Haloperidol decanoate-treated animals, acting as controls for possible antipsychotic effects, did not have significantly altered neuronal EAAT2b mRNA levels. The novel finding that EAAT2b levels are increased in populations of anterior cingulate cortex pyramidal cells further demonstrates a role for neuronal glutamate transporter splice variant expression in schizophrenia.

Milk fat globule membrane isolated from buttermilk or whey cream and their lipid components inhibit infectivity of rotavirus in vitro.

Milk fat is encapsulated in a milk fat globule membrane (MFGM) that contains bioactive glycoproteins and glycolipids. The MFGM inhibits infectivity of rotavirus (RV), activity that has been attributed to its glycoprotein and carbohydrate components. However, previous studies of proteins and oligosaccharides in the MGFM have not accounted for all the bioactivity associated with the complete MFGM. The lipid fraction of the MFGM accounts for half of its composition by weight, and we postulate that this fraction should be tested by itself to determine if it plays a role in antiviral activity. Herein, the anti-RV activity of an organic extract of MFGM was tested. Natural and whey buttermilk powders containing bovine MFGM enriched in polar lipids were prepared by microfiltration and supercritical fluid extraction treatment to reduce the triglyceride content of the powders. Lipid fractions were then extracted from the MFGM using both single- and dual-phase extraction methods. Whole MFGM and organic extracts were screened in MA-104 cells for anti-infective activity against a neuraminidase-sensitive rotavirus using a focus-forming unit assay. Dose-dependent inhibition was observed for whole buttermilk and cheese whey MFGM against the rotavirus. In general, buttermilk MFGM exhibited greater RV percentage inhibition than cheese whey MFGM. Organic-soluble anti-RV compounds were identified in bovine MFGM. The most active fraction, isolated by dual-phase extraction and iatrobead chromatography, was free of proteins and highly nonpolar. Further separation of this fraction in a less polar solvent (30:1 chloroform:methanol) resolved at least 5 lipid-containing compounds, which likely contribute to the anti-RV activity associated with bovine MFGM. In summary, lipid components associated with MFGM appear to contribute in large part to the anti-RV activity associated with the bovine MFGM.

Modelling the behaviour of mechanical biological treatment outputs in landfills using the GasSim model.

The pretreatment of the biodegradable components of municipal solid waste (MSW) has been suggested as a method of reducing landfill gas emissions. Mechanical biological treatment (MBT) is the technology being developed to provide this reduction in biodegradability, either as an alternative to source segregated collection or for dealing with residual MSW which still contains high levels of biodegradable waste. The compost like outputs (CLOs) from MBT plants can be applied to land as a soil conditioner; treated to produce a solid recovered fuel (SRF) or landfilled. In this study the impact that landfilling of these CLOs will have on gaseous emissions is investigated. It is important that the gas production behaviour of landfilled waste is well understood, especially in European member states where the mitigation of gaseous emissions is a legal requirement. Results of an experiment carried out to characterise the biodegradable components of pretreated biowastes have been used with the GasSim model to predict the long term emissions behaviour of landfills accepting these wastes, in varying quantities. The landfill directive also enforces the mitigation of potential methane emissions from landfills, and the ability of landfill operators to capture gaseous emissions from low emitting landfills of the future is discussed, as well as new techniques that could be used for the mitigation of methane generation.

Biochemical and morphological developments are partially impaired in intestinal mucosa from growing pigs fed reduced-protein diets supplemented with crystalline amino acids.

The objective of this study was to determine if a reduction in dietary CP, with partial replacement of the intact protein with crystalline AA (CAA), would alter growth, morphology, and free or peptide-bound AA concentrations of intestinal mucosa in growing pigs. Twenty-four barrows (37.0 +/- 1.5 kg of BW) were fed 1 of 4 diets for 24 d: 16.1% CP with no CAA, or 12.8, 10.1, or 7.8% CP (analyzed values, as-fed) containing CAA. As CP decreased, CAA were gradually increased to meet requirements on a true ileal digestible basis. Pigs were euthanized 2 h postmeal on d 24, and mucosal samples from duodenum, jejunum, and ileum were collected. Reducing dietary CP decreased ADG, G:F, and final weight (linear, P < 0.05). With reduced dietary CP, mucosal protein concentration decreased in the jejunum (quadratic, P < 0.05) and tended to decrease in the ileum (linear, P = 0.062). Reduction of the dietary CP concentration from 16.1 to 7.8% tended to decrease the crypt depth (linear, P < 0.10) and decreased villus width (linear, P < 0.05) in duodenum and jejunum mucosa but did not reduce villus height or villus surface area in any regions of the small intestine. In the duodenum, a reduction in dietary CP increased free Lys, Met, and Thr (linear, P < 0.05) and peptide-bound Lys and Thr (quadratic, P < 0.10). In the jejunum, reducing CP decreased free Cys (linear P < 0.05) and tended to decrease free Asn and His (linear, P < 0.10) and peptide-bound His (quadratic, P = 0.061) and Ile, Leu, and Val (linear, P < 0.10). In the ileum, reducing CP decreased free Asn, Ser, Tyr, Arg, His, Phe (linear, P < 0.05), and Leu (linear, P = 0.054) and peptide-bound Gly and Ser (linear, P < 0.05) and tended to decrease peptide-bound Ile, Leu, Phe, Val (linear, P < 0.10), and Lys (linear P < 0.05). In conclusion, reduced-CP diets supplemented with CAA lead to a reduction in growth performance, associated with biochemical and morphological modifications of the intestinal mucosa.

Effect of a short-term fast on intestinal disaccharidase activity and villus morphology of piglets suckling insulin-like growth factor-I transgenic sows.

The objectives of this study were to use transgenic sows that overexpress IGF-I in milk to investigate the effect of a short-term fast on piglet intestinal morphology and disaccharidase activity and to determine how milk-borne IGF-I influences the response to fasting. After farrowing, litters were normalized to 10 piglets. On d 6, piglets (n = 30) suckling IGF-I transgenic (TG) sows and piglets (n = 30) suckling nontransgenic sows (control) were assigned randomly to three treatments: fed piglets (0 h), which remained with the sow until euthanized on d 7, or fasted piglets, which were removed from the sow at either 6 or 12 h before euthanasia on d 7. Serum IGF-I and IGFBP, intestinal weight and length, jejunal protein and DNA content, disaccharidase activity, and villus morphology were measured. Fasting for 12 h resulted in a negative weight change between d 6 and 7 (quadratic response to fasting; P < 0.001). Piglets suckling TG sows tended to have greater intestinal length (P = 0.068), but no effect of IGF-I overexpression was noted for intestinal weight. Fasting, however, resulted in linear (P < 0.001) and quadratic (P = 0.002) decreases in intestinal weight. Serum IGF-I did not differ between control and TG sows, but decreased linearly (P = 0.003) with fasting. Serum IGFBP-4 decreased (linear and quadratic; P < or = 0.02) with fasting, whereas IGFBP-1 increased quadratically (P < 0.001) with fasting. Jejunal villus height, width, and crypt depth were all increased with fasting (linear and quadratic; P < 0.04). Disaccharidase activity was not affected by fed state; however, piglets suckling TG sows had greater jejunal lactase-phlorhizin hydrolase (P < 0.01) and sucrase-isomaltase (P = 0.02) activities than control piglets. In summary, intestinal weight, villus morphology, serum IGF-I, serum IGFBP-1 and -4, and piglet BW change were altered (P < or = 0.02) in response to fasting. Thus, the duration of food deprivation before euthanization should be considered when designing experiments to assess intestinal development or the IGF axis, as the magnitude of differences between the fed and fasted state may exceed those expected as a result of experimental treatment.

Effects of protein deprivation on subsequent growth performance, gain of body components, and protein requirements in growing pigs.

Forty-eight barrows were used in a 2 x 6 factorial arrangement to test a hypothesis that feeding a protein-deficient diet affects subsequent growth response by altering the efficiency of protein utilization. Barrows were individually fed either a 9% crude protein (CP) diet or an 18% CP diet from 20 to 30 kg of body weight (BW) (depletion phase). From 30 to 45 kg BW (realimentation phase), pigs were fed one of six experimental diets with CP levels of 11.8, 13.1, 14.3, 15.6, 18.8, and 21.8%. Four pigs were slaughtered at 20 kg BW to determine initial body composition. Four pigs from each treatment in depletion phase (a total of eight) were slaughtered at 30 kg BW, and all pigs from each treatment in realimentation phase (a total of 36) were slaughtered at 45 kg BW for subsequent compositional analysis. Pigs were bled at 20, 30, and 40 kg BW for blood urea nitrogen (BUN), insulin-like growth factor (IGF)-I, and IGF-binding protein (IGFBP) assays. Pigs were given three times the maintenance digestible energy requirement (3 x 120 kcal BW(-0.75) x d(-1)) in three equal meals daily. The feed allowance was adjusted every 3 d. During the depletion phase, pigs fed the 18% CP diet grew faster and more efficiently (P < 0.01) and gained more (P < 0.01) water and protein than did pigs fed the 9% CP diet. Pigs fed the 18% CP diet showed higher (P < 0.01) BUN values, IGF-I concentrations, and IGFBP ratios than pigs fed the 9% CP diet. During the realimentation phase, pigs fed the 9% CP diet during the depletion phase grew faster (P < 0.05), tended to grow more efficiently (P = 0.066), gained more water (P < 0.01), and tended to gain more protein (P = 0.068) than pigs fed the 18% CP diet during the depletion phase. Pigs fed the 9% CP diet during the depletion phase tended (P = 0.069) to have a higher protein requirement during the realimentation phase than pigs fed the 18% CP diet during the depletion phase. When measured at 40 kg BW, pigs fed the 9% CP diet had a lower (P < 0.05) BUN than pigs fed the 18% CP diet during the depletion phase. However, the plasma IGF-I concentration and IGFBP ratio at 40 kg BW were not affected by dietary CP level fed during the depletion phase. This study indicates that pigs fed a protein-deficient diet exhibit compensatory growth. During the period of compensatory growth, the requirement of CP for those pigs is higher than that of pigs previously fed an adequate diet. This study also suggests BUN can be used as an indicator of protein utilization efficiency and compensatory growth.

Acidomucin goblet cell expansion induced by parenteral nutrition in the small intestine of piglets.

Total parenteral nutrition (TPN) impairs small intestine development and is associated with barrier failure, inflammation, and acidomucin goblet cell expansion in neonatal piglets. We examined the relationship between intestinal goblet cell expansion and molecular and cellular indices of inflammation in neonatal piglets receiving TPN, 80% parenteral + 20% enteral nutrition (PEN), or 100% enteral nutrition (control) for 3 or 7 days. Epithelial permeability, T cell numbers, TNF-alpha and IFN-gamma mRNA expression, and epithelial proliferation and apoptosis were compared with goblet cell numbers over time. Epithelial permeability was similar to control in the TPN and PEN jejunum at day 3 but increased in the TPN jejunum by day 7. By day 3, intestinal T cell numbers were increased in TPN but not in PEN piglets. However, goblet cell expansion was established by day 3 in both the TPN and PEN ileum. Neither TNF-alpha nor IFN-gamma mRNA expression in the TPN and PEN ileum correlated with goblet cell expansion. Thus goblet cell expansion occurred independently of overt inflammation but in association with parenteral feeding. These data support the hypothesis that goblet cell expansion represents an initial defense triggered by reduced epithelial renewal to prevent intestinal barrier failure.

Intestinal protein and LPH synthesis in parenterally fed piglets receiving partial enteral nutrition and enteral insulinlike growth factor 1.

BACKGROUND: Providing partial enteral nutrition (PEN) supplemented with insulinlike growth factor-1 (IGF-1) to parenterally fed piglets increases lactase-phlorizin hydrolase (LPH) activity, but not LPH mRNA. The current aim was to investigate potential mechanisms by which IGF-1 up-regulates LPH activity. METHODS: Newborn piglets (n = 15) received 100% parenteral nutrition (TPN), 80% parenteral nutrition + 20% parenteral nutrition (PEN), or PEN + IGF-1 (1.0 mg. kg-1. d-1) for 7 days. On day 7, [2H3]-leucine was intravenously administered to measure mucosal protein and brush border LPH (BB LPH) synthesis. RESULTS: Weight gain, nutrient intake, and jejunal weight and length were similar among the treatment groups. Partial enteral nutrition alone increased mucosal weight, villus width and cross-sectional area, LPH activity, mRNA expression, and high mannose LPH precursor (proLPHh) abundance compared with TPN (P<0.05). Insulinlike growth factor-1 further increased mucosal weight, LPH activity, and LPH activity per unit BB LPH approximately twofold over PEN alone (P < 0.05) but did not affect LPH mRNA or the abundance of proLPHh (one of the LPH isoforms) or mature LPH. Isotopic enrichment of [2H3]-leucine in plasma, mucosal protein, and LPH precursors, and the fractional and absolute synthesis rates of mucosal protein and LPH were similar among the treatment groups. Insulinlike growth factor-1 treatment increased total mucosal protein synthesis (60%, P < 0.05) but not LPH synthesis compared with the other two groups. CONCLUSIONS: Because IGF-1 did not affect the fractional synthesis rate of either mucosal protein or LPH, the authors suggest that enteral IGF-1 increases mucosal protein mass and LPH activity by suppressing mucosal proteolytic degradation.

Oral IGF-I alters the posttranslational processing but not the activity of lactase-phlorizin hydrolase in formula-fed neonatal pigs.

To determine the cellular mechanism whereby oral insulin-like growth factor I (IGF-I) increases intestinal lactase-phlorizin hydrolase (LPH) activity, we studied 2-d-old pigs fed cow's milk formula (control, n = 5), formula + low IGF-I (0.5 mg/L; n = 6) or formula + high IGF-I (12.0 mg/L, n = 6) for 15 d. On d 15, intestinal protein synthesis and lactase processing were measured in vivo in fed pigs using a 6-h intravenous, overlapping infusion of multiple stable isotopes (2H(3)-Leu, 13C(1)-Leu, 13C(1)-Phe, 2H(5)-Phe, 13C(6)-Phe and 13C(9)-Phe). Morphometry and cell proliferation also were measured in the jejunum and ileum. Neither dose of IGF-I affected the masses of wet tissue, protein or DNA, or the villus height, cell proliferation or LPH-specific activity. Oral IGF-I decreased the synthesis and abundance of prolactase-phlorizin hydrolase (pro-LPH), but increased brush-border (BB)-LPH synthesis in the ileum. The BB-LPH processing efficiency was twofold to threefold greater in IGF-fed than in control pigs. In all pigs, villus height and the total mucosal and specific activity of LPH activity were greater in the ileum than in the jejunum, yet the synthesis of BB-LPH were significantly lower in the ileum than in the jejunum. We conclude that oral IGF-I increases the processing efficiency of pro-LPH to BB-LPH but does not affect LPH activity. Moreover, the posttranslational processing of BB-LPH is markedly lower in the ileum than in the jejunum.

Testosterone and food restriction modulate hepatic lycopene isomer concentrations in male F344 rats.

We previously demonstrated that the castration of male rats profoundly increases hepatic lycopene compared with intact controls. Here we further characterized the role of testosterone in modulating hepatic lycopene accumulation and isomer patterns in male rats. Furthermore, because castration significantly decreases ad libitum food consumption, we investigated the influence of food restriction on lycopene metabolism. Forty male F344 rats 8 wk of age were randomly assigned to one of four treatments (n = 10/group): 1) intact, free access to food, 2) castration, free access to food, 3) castration plus testosterone implants, free access to food and 4) intact, 20% food restricted. All rats were fed an AIN-based diet with 0.25 g lycopene (as 10% water-soluble beadlets)/kg diet for 3 wk. Serum testosterone was 5.31 +/- 1.46 nmol/L in intact controls allowed free access to food, reduced in castrated animals (0.52 +/- 0.10, P < 0.0001 versus controls) and intact, food-restricted rats (1.53 +/- 0.49 nmol/L, P < 0.0001 versus controls) and greater (17.23 +/- 3.09 nmol/L) in castrated rats administered testosterone (P < 0.0001 versus controls). Castrated rats accumulated approximately twice as much liver lycopene (74.5 +/- 8.5 nmol/g; P < 0.01 versus controls) as intact rats allowed free access to food (39.5 +/- 5.0) despite 13% lower dietary lycopene intake (P < 0.001; 3.38 +/- 0.07 versus 3.95 +/- 0.06 mg lycopene/d). Testosterone replacement in castrated rats returned liver lycopene concentrations (32.5 +/- 5.5 nmol lycopene/g with 3.76 +/- 0.05 mg dietary lycopene/d) to those observed in intact rats. Food restriction resulted in a 20% decrease in lycopene intake but significantly increased liver lycopene by 68% (66.3 +/- 7.9 nmol lycopene/g with 3.38 +/- 0.00 mg lycopene/d) compared with controls and castrated rats administered testosterone. These results suggest that androgen depletion and 20% food restriction increase hepatic lycopene accumulation. We hypothesize an endocrine and dietary interaction, where higher androgen concentrations and greater energy intake may stimulate lycopene metabolism and degradation.

Time course changes in IGFBP-1 after treadmill exercise and postexercise food intake in rats.

Prolonged exercise increases circulating insulin-like growth factor binding protein-1 (IGFBP-1) in humans and animals, but its physiological significance is unknown. This study examined 1) time-course changes in plasma IGFBP-1 and hepatic IGFBP-1 mRNA expression after exercise, 2) changes in IGFBP-1 in relation to plasma glucose, insulin, and IGF-I, and 3) the impact of feeding a postexercise meal on the IGFBP-1 response. Food-deprived male rats were vigorously run on a treadmill and compared with nonexercised controls at 15 min and 1, 4, 8, and 12 h after exercise. Circulating insulin concentrations in exercised rats were lower than in controls at 15 min and 1 h, whereas plasma glucose and IGF-I remained unaffected. Circulating and hepatic expression of IGFBP-1 was markedly increased above that of controls at 15 min, 1 h, and 12 h. In a separate experiment, one-half of the exercised animals received a nutritionally complete meal immediately after the experimental run. The meal elevated plasma insulin and glucose concentrations at 15 min and 1 h. Despite this change in nutritional status, serum IGFBP-1 concentrations and hepatic IGFBP-1 abundance remained elevated at 15 min and 1 h. These results demonstrate that the IGFBP-1 response to a single bout of treadmill exercise is short in duration and independent of insulin, glucose, and amino acid availability.

Transgenic alteration of sow milk to improve piglet growth and health.

There are many potential applications of transgenic methodologies for developing new and improved strains of livestock. One practical application of transgenic technology in pig production is to improve milk production or composition. The first week after parturition is the period of greatest loss for pig producers, with highest morbidity and mortality attributed to malnutrition and scours. Despite the benefits to be gained by improving lactation performance, little progress has been made in this area through genetic selection or nutrition. Transgenic technology provides an important tool for addressing the problem of low milk production and its detrimental impact on pig production. Transgenic pigs over-expressing the milk protein bovine alpha-lactalbumin were developed. alpha-Lactalbumin was selected for its role in lactose synthesis and regulation of milk volume. Sows hemizygous for the transgene produced as much as 0.9 g bovine alpha-lactalbumin l-1 pig milk. The outcomes assessed were milk composition, milk yield and piglet growth. First parity alpha-lactalbumin gilts had higher milk lactose content in early lactation and 20-50% greater milk yield on days 3-9 of lactation than did non-transgenic gilts. Weight gain of piglets suckling alpha-lactalbumin gilts was greater (days 7-21 after parturition) than that of control piglets. Thus, transgenic over-expression of milk proteins may provide a means for improving the lactation performance of pigs.

Perturbations in factors that modulate osteoblast functions in vitamin B6 deficiency.

It was hypothesized that the widespread structural defect of collagen in connective tissue of vitamin B6 deficient-animals and the consequent alteration in bone biomechanical properties cause an additional stress to their inflamed swollen tibiotarsometatarsal joints. The present study showed a 32% elevation (P < 0.02) in mean plasma free cortisol concentration. Vitamin D metabolism was impaired but without changing plasma calcium homeostasis and bone mineral content. Mean plasma calcitriol [1,25(OH)2D] concentration was significantly reduced (P < 0.001). Because plasma calcidiol concentration did not change, we speculated that either renal 25-hydroxycalciferol-1alpha-hydroxylase activity was reduced or 1,25(OH)2D turnover was increased. Plasma osteocalcin, an index of osteoblast function related to bone formation, was significantly decreased (P < 0.05). This adverse effect on osteoblasts was consistent with the reduction of bone specific alkaline phosphatase activity (another index of bone formation) found in a previous study. The excess of cortisol may have impaired these bone cells functions directly and (or) indirectly via the decline in calcitriol synthesis. Plasma hydroxyproline concentrations in B6-deficient animals were found to be significantly reduced (P < 0.001), suggesting that cortisol in excess had also a suppressive effect on another hydroxylase, namely tissue (mainly bone and liver) prolyl hydroxylase. The bone uncoupling (in formation and resorption) associated with vitamin B6 deficiency seems to be due to secondary hypercortisolism and (or) another unknown factors but not related to a change in bone modulators such as IGF-1 and eicosanoids.

Investigation of three doses of oral insulin-like growth factor-I on jejunal lactase phlorizin hydrolase activity and gene expression and enterocyte proliferation and migration in piglets.

In a previous study, oral IGF-I at 65 nM increased lactase phlorizin hydrolase (LPH) activity and villus height in piglets, however, the mechanisms were unknown. Herein, the response to a range of doses of IGF-I was investigated and we hypothesized that LPH and villus height would respond to oral IGF-I in a dose-dependent manner. Two 14-d experiments were conducted using cesarean-derived piglets. In experiment 1, piglets (n = 28) were fed formula containing 0, 33, 65, or 131 nmol/L (0, 0.25, 0.5, or 1.0 mg/L) recombinant human IGF-I. In experiment 2, 5'-bromodeoxyuridine was administered to piglets fed formula alone (n = 4) or containing 131 nmol/L IGF-I (n = 4). IGF-I did not affect body weight gain or intestinal weight or length. Jejunal villus height and LPH activity were significantly greater in piglets fed 131 nmol IGF-I/L than control piglets. Villus height and lactase activity in piglets fed the 33 and 65 nmol/L IGF-I doses were similar and intermediate between control and 131 nmol IGF-I/L. Jejunal mRNA expression and LPH polypeptide abundance were investigated in piglets receiving 0 or 131 nmol/L IGF-I. Steady state LPH mRNA abundance was significantly higher (p < 0.05) in IGF-I-treated piglets. The relative abundance of proLPH(h) was not significantly increased (p = 0.06) by IGF-I treatment. Mucosal DNA content and DNA synthesis were greater in piglets receiving 131 nmol IGF-I/L than control, however, enterocyte migration and mucosal protein content were unaffected. Thus, oral IGF-I increased jejunal LPH activity and LPH mRNA abundance and stimulated intestinal cell hyperplasia in normal piglets.

Dietary fat during pregnancy and lactation increases milk fat and insulin-like growth factor I concentrations and improves neonatal growth rates in swine.

Primiparous (n = 24) and multiparous (n = 24) sows were used to examine the effects of supplemental dietary fat and induction of parturition (d 112) on colostrum and milk composition and suckling piglet growth. Sows were assigned to one of eight treatments on d 90 of gestation that included variables such as parity (1 vs. >/=3), dietary fat (0 vs. 10%), and farrowing (natural vs. induction via lutalyse on d 112). Piglets suckling fat-supplemented dams grew up to 25% faster than control pigs nursing unsupplemented sows (250 vs. 200 g/d; P < 0.01). Improved growth was correlated with elevated milk fat and insulin-like growth factor (IGF) concentrations associated with fat supplementation. Dietary fat elevated milk fat concentration at 48 and 72 h postfarrowing by 21.6 and 22.6%, respectively (P < 0.05). Compared with nonfat-fed controls, multiparous sows fed 10% fat showed a more consistent rise in milk fat concentration, with 26% and 41% elevations for induced or naturally farrowing sows, respectively, vs. a 19% reduction or a 1% elevation in induced or naturally farrowing gilts (P < 0.01). The concentration of milk IGF-I tended to be lower in gilts than in multiparous sows (P < 0.2, 95.7 vs. 117.4 microg/L), and levels were particularly low in milk from induced gilts receiving no additional dietary fat (44.7 microg/L). However, fat supplementation elevated IGF-I to levels (110.6 microg/L) exceeding those measured in unsupplemented, naturally farrowing control sows and gilts (95.8 microg/L). In conclusion, supplemental dietary fat elevates milk fat in multiparous sows more than primiparous gilts regardless of farrowing treatment (induced vs. natural farrowing) and improves piglet growth throughout lactation irrespective of parity or farrowing treatment. The potential of supplemental dietary fat to reverse the reductions in milk IGF-I observed in first-parity females and in dams induced to farrow merits further investigation.

Total parenteral nutrition alters molecular and cellular indices of intestinal inflammation in neonatal piglets.

BACKGROUND: The adverse effects of TPN on systemic immunity are well-documented; however, the impact of IV feeding on neonatal intestinal immunity is unknown. METHODS: A piglet TPN model was used to compare immune cell composition within the intestinal epithelium and lamina propria of parenterally and orally fed piglets. RESULTS: Small intestinal weight of piglets maintained intravenously was reduced 50% after 7 days. Intestinal atrophy in piglets fed parenterally was evidenced by decreased width of intestinal villi and colon cuffs and reduced intestinal crypt depth. The numbers of CD4+ and CD8+ T lymphocytes were threefold greater within the lamina propria of jejunal and ileal villi of piglets supported intravenously. Inverse correlations were observed between villus height or width and T-lymphocyte numbers (r = -.80; p < .05). Major histocompatibility complex class II mRNA expression, an indicator of localized inflammation, was increased in the ileum and colon of piglets receiving parenteral nutrition. Goblet cell numbers were two-fold greater in jejunal and ileal villi, and mast cells were more abundant in the colon of piglets fed parenterally. Furthermore, jejunal T-lymphocyte numbers were correlated with goblet cell numbers (r = .80; p = .01). CONCLUSIONS: These data identify molecular and cellular indices of intestinal inflammation that are responsive to IV feeding in neonates and provide a novel framework to investigate mechanisms underlying gut atrophy during TPN.