Clinical features and magnesium levels: Novel insights in 15q11.2 BP1-BP2 copy number variants.

BACKGROUND: Investigating copy number variations (CNVs) such as microdeletions or microduplications can significantly contribute to discover the aetiology of neurodevelopmental disorders. 15q11.2 genomic region, including NIPA1 and NIPA2 genes, contains a recurrent but rare CNV, flanked by the break points BP1 and BP2. Both BP1-BP2 microdeletion and microduplication have been associated with intellectual disability (ID), neuropsychiatric/behavioural disturbances and mild clinical features, even if with incomplete penetrance and variable expressivity. The pathogenic role of this CNV is quite unclear though. Unknown variants in other DNA regions and parent-of-origin effect (POE) are some of the mechanisms that have been proposed as an explanation of the wide phenotypic variability. As NIPA1 and NIPA2 encode for proteins that mediate magnesium (Mg2+ ) metabolism, it has been suggested that urinary Mg2+ levels could potentially represent informative and affordable biomarkers for a rapid screening of 15q11.2 duplications or deletions. Furthermore, magnesium supplementation has been proposed as possible therapeutic strategy. METHODS: Thirty one children with ID and/or other neurodevelopmental disorders carrying either a duplication or a deletion in 15q11.2 BP1-BP2 region have been recruited. When available, blood samples from parents have been analysed to identify the CNV origin. All participants underwent family and medical data collection, physical examination and neuropsychiatric assessment. Electroencephalogram (EEG) and brain magnetic resonance imaging (MRI) scan were performed in 15 children. In addition, 11 families agreed to participate to the assessment of blood and urinary Mg2+ levels. RESULTS: We observed a highly variable phenotypic spectrum of developmental issues encompassing ID in most subjects as well as a variety of behavioural disorders such as autism and attention-deficit disorder/attention-deficit hyperactivity disorder. Dysmorphic traits and malformations were detected only in a minority of the participants, and no clear association with growth anomalies was found. Abnormal brain MRI and/or EEG were reported respectively in 64% and 92% of the subjects. Inheritance assessment highlighted an excess of duplication of maternal origin, while cardiac alterations were detected only in children with 15q11.2 CNV inherited from the father. We found great variability in Mg2+ urinary values, without correlation with 15q11.2 copy numbers. However, the variance of urinary Mg2+ levels largely increases in individuals with 15q11.2 deletion/duplication. CONCLUSIONS: This study provides further evidence that 15q11.2 BP1-BP2 CNV is associated with a broad spectrum of neurodevelopmental disorders and POE might be an explanation for clinical variability. However, some issues may question the real impact of 15q11.2 CNV on the phenotype in the carriers: DNA sequencing could be useful to exclude other pathogenic gene mutations. Our results do not support the possibility that urinary Mg2+ levels can be used as biomarkers to screen children with neurodevelopmental disorders for 15q11.2 duplication/deletion. However, there are evidences of correlations between 15q11.2 BP1-BP2 CNV and Mg2+ metabolism and future studies may pave the way to new therapeutic options.

Effects of dietary menhaden oil on growth and reproduction in gilts farrowed by sows that consumed diets containing menhaden oil during gestation and lactation.

In sows, n-3 fatty acids increase litter sizes, however, effects on gilt reproductive development have not been adequately studied. Moreover, not determined are effects of feeding n-3 fatty acids to sows on reproduction in offspring. The objective here was to determine effects of 4% dietary menhaden oil on growth and puberty in gilts farrowed by sows fed menhaden oil. Sows (n = 44) were assigned to: (1) control gestation and lactation diets, or (2) diets including menhaden oil. For primiparous sows only, total litter size and born alive were greater (P < 0.05) in females fed menhaden oil. Conversely, pigs from primiparous controls were heavier (P < 0.05) than pigs from primiparous sows fed menhaden oil (parity by diet interactions, P < 0.01). Diet did not affect (P > 0.20) other sow and litter characteristics. At weaning, 84 gilts from control- or menhaden oil sows were placed three gilts per pen and provided control diets or diets containing menhaden oil. Nursery and grow-finish feed intake and feed efficiency were similar (P > 0.21) for gilts from the different sows and weight gain was similar (P > 0.24) for gilts fed control or menhaden diets. Gilts fed menhaden oil tended to eat less in the nursery (1.18±0.08 kg v. 0.98±0.08 kg; P = 0.09) and overall (1.83±0.04 kg v. 1.72±0.04 kg; P = 0.06). Thus, overall feed to gain was greater (2.52±0.03 v. 2.33±0.03; P < 0.01) and nursery (2.12±0.04 v. 1.80±0.04; P = 0.10) and grow-finish (3.07±0.19 v. 2.58±0.19; P = 0.08) feed to gain tended to be greater, for control gilts. Age at puberty was greater (P = 0.02) for gilts from menhaden oil-fed sows (205.1±3.2 days) compared to gilts from controls (193.9±3.2 days) and tended to be greater (P = 0.09), for controls (203.5±3.2 days) compared to gilts fed menhaden oil (195.5±3.2 days). A tendency existed (P = 0.09) for greater follicular fluid in gilts fed menhaden oil, however, ovulation rate and ovarian, luteal and uterine weights were not affected by sow diet, gilt diet or the interaction (P > 0.23). Feeding gilts menhaden oil enhanced feed efficiency and hastened puberty onset. Gilts from sows consuming menhaden oil exhibited delayed puberty and retaining females from sows fed this feedstuff may be ill advised.

Supplementing antioxidants to pigs fed diets high in oxidants: II. Effects on carcass characteristics, meat quality, and fatty acid profile.

The study was conducted to determine effects of dietary supplementation with a blend of antioxidants (ethoxyquin and propyl gallate) on carcass characteristics, meat quality, and fatty acid profile in finishing pigs fed a diet high in oxidants. A total of 100 crossbred barrows (10.9±1.4 kg BW, 36±2 d of age) were randomly allotted to 5 diet treatments (5 replicate pens per treatment, 4 pigs per pen). Treatments included: 1) HO: high oxidant diet containing 5% oxidized soy oil and 10% PUFA source which contributed 5.56% crude fat and 2.05% docosahexanoic acid (DHA) to the diet; 2) VE: the HO diet with 11 IU/kg of added vitamin E; 3) AOX: the HO diet with antioxidant blend (135 mg/kg); 4) VE+AOX: the HO diet with both vitamin E and antioxidant blend; and 5) SC: a standard corn-soy control diet with nonoxidized oil and no PUFA source. The trial lasted for 118 d; on d 83, the HO diet pigs were switched to the SC diet due to very poor health. From that point, the VE pigs displayed the poorest performance. On d 118, 2 pigs from each pen were harvested for sampling. Compared to pigs fed SC diet, the HO and VE pigs (P<0.05) showed lighter carcass weight, less back fat, less lean body mass, and smaller loin eye area. In addition, the VE pigs had decreased dressing percentage than the AOX and VE+AOX pigs (65.7 vs. 75.3 and 74.2%). Compared to the SC pigs, greater moisture percentage (74.7 vs. 77.4%) and less extractable lipid content (2.43 vs. 0.95%) were found in VE fed pigs (P<0.05). Drip loss of loin muscle in VE pigs was less than SC pigs (0.46 vs. 3.98%, P=0.02), which was associated with a trend for a greater 24-h muscle pH (5.74 vs. 5.54, P=0.07). The antioxidant blend addition in the high oxidant diet attenuated all of these effects to levels similar to SC (P>0.05), except a* value (redness) and belly firmness. Visible yellow coloration of backfat and lipofuscin in HO and VE pigs was observed at harvest at d 118. The high oxidant diet resulted in greater concentration of DHA in backfat (P<0.001); switching the diet on d 83 resulted in HO pigs having a similar fatty acid profile to SC at d 118 pigs. Vitamin E concentration in plasma and muscle was greater in HO and SC than VE, AOX, and VE+AOX on d 118. Feeding the high oxidant diet caused a series of changes in carcass characteristics and meat quality. Addition of antioxidant blend attenuated many of these, whereas the protective effects of supplemental vitamin E at 11 IU/kg were minimal during the finisher phase of the study.

Supplementing antioxidants to pigs fed diets high in oxidants: I. Effects on growth performance, liver function, and oxidative status.

The objective of the study was to determine the effects of a dietary antioxidant blend (ethoxyquin and propyl gallate) and vitamin E on growth performance, liver function, and oxidative status in pigs fed diets high in oxidants. Crossbred barrows (n=100, 10.91±0.65 kg BW, 36±2 d of age, Landrace×Duroc) were allotted to 5 treatments on the basis of BW (5 replicate pens per treatment, 4 pigs per pen). Treatments included 1) HO, high-oxidant diet containing 5% oxidized soybean oil and 10% PUFA source (providing 2.05% docosahexaenoic acid in the diet), 2) VE, the HO diet with 11 IU/kg of added vitamin E, 3) AOX, the HO diet with antioxidant blend (135 mg/kg), 4) VE+AOX, the HO diet with both vitamin E and antioxidant blend, and 5) SC, a standard corn-soy control diet. The trial lasted for 118 d; on d 83, the HO diet pigs were switched to the SC diet because the animals were displaying very poor health. Compared with SC pigs, HO pigs had decreased ADG (0.92 vs. 0.51 kg for d 26 to 55, 1.29 vs. 0.34 kg for d 56 to 82; P<0.05) and ADFI (1.84 vs. 0.96 kg for d 26 to 55, 3.41 vs. 1.14 kg for d 56 to 82; P<0.05). However, switching the HO pigs to the SC diet resulted in HO pigs having a greater ADG than VE-fed pigs from d 83 to 118 (0.90 vs. 0.60 kg; P<0.05). The antioxidant blend restored pig performance to a level similar that of pigs fed the SC diet (P>0.05) with greater G:F for the entire period (0.44 vs. 0.38; P<0.05). A greater liver to BW ratio was found in HO compared with other treatments on d 55 and in VE on d 118. Total bilirubin concentration in plasma of HO pigs on d 55 was greater than that in VE+AOX pigs (P<0.05), whereas on d 118, bilirubin concentration in VE was higher than those in VE+AOX and SC (P<0.05). A similar trend was observed in aspartate transaminase. Plasma concentrations of thiobarbituric acid reactive substances (TBARS) and carbonyl were elevated (P<0.05) in the HO pigs compared with the SC pigs on d 55 but not on d 118. Liver TBARS and carbonyl concentrations showed a similar trend, except that HO pigs had the greatest carbonyl concentration on d 118. Pigs fed AOX diets had plasma and liver TBARS and carbonyl concentrations similar to those fed SC diets. In the oxidative stress model used in this study, dietary addition of antioxidant blend or antioxidant blend+vitaimin E was effective in improving growth, liver function, and plasma markers of oxidative stress, but VE alone was not.

Performance and physiology of pigs administered spray-dried plasma protein during the late suckling period and transported after weaning.

The objective was to determine the effects of spray-dried plasma protein (SDPP), given as an oral gavage during the last 5 d of suckling, on weight gain and physiology in pigs after weaning and transportation for 5 h. Pigs were assigned to 1 of 4 treatments: 1) SDPP (9.375 g) + transportation, 2) water + transportation, 3) SDPP + no transportation, and 4) water + no transportation (n = 10 barrows and 10 gilts per treatment). Pigs received 25 mL of the SDPP (0.375 g/mL) or water twice daily. There was no effect (P = 0.55) of gavage on weaning BW. On the day of weaning, BW decreased in all groups but the magnitude was greatest in SDPP pigs that were transported (gavage × transportation × time, P = 0.03). Rectal temperatures increased in all groups but were greater after transportation than after no transportation (gavage × transportation × time, P < 0.01). Effects of transportation × time existed for several blood chemistry measures. Urea and protein concentrations increased (P < 0.01) in transported pigs only. Creatinine, chloride, and albumin increased (P < 0.01) and CO2 decreased (P < 0.01) in both transported and nontransported pigs, but the magnitudes of change were greater after transportation. Concentrations of sodium increased (P < 0.01) only in transported pigs receiving water and not in the other groups (gavage × transportation × time, P < 0.01). Concentrations of phosphorous (P < 0.01) were affected by sex × gavage × transportation × time and increased (P < 0.01) in transported, water-treated gilts but not barrows. Overall changes in concentrations of urea, creatinine, chloride, CO2, protein, albumin, sodium, and phosphorous are consistent with dehydration in transported pigs in this study and in the case of sodium (both sexes) and phosphorous (gilts only), these minerals were maintained by prior gavage with SDPP. Transported pigs receiving SDPP tended (P = 0.1) to have greater concentrations of glucose than transported pigs receiving water and had similar glucose levels to nontransported pigs receiving water, suggesting that SDPP before weaning and transportation helps to maintain concentrations. Postweaning BW was affected (P = 0.01) by gavage × time and at wk 5, pigs gavaged with SDPP tended (P = 0.1) to weigh more than pigs gavaged with water. Providing SDPP before weaning prevented transportation-induced changes in some blood components and enhanced postweaning weight gain.

Effects of dietary supplementation with an organic source of selenium on characteristics of semen quality and in vitro fertility in boars.

Semen characteristics in boars fed organic or inorganic sources of Se were assessed in 3 experiments. Crossbred boars were randomly assigned at weaning to 1 of 3 dietary treatments: I) basal diets with no supplemental Se (control), II) basal diets with 0.3 mg/kg of supplemental Se from an organic source (Sel-Plex, Alltech Inc., Nicholasville, KY), and III) basal diets supplemented with 0.3 mg/kg of supplemental Se from sodium selenite (Premium Selenium 270, North American Nutrition Co. Inc., Lewisburg, OH). For Exp. 1, semen was collected from boars (n = 10/dietary treatment) on 5 consecutive days at 15 mo of age. Effects of treatment × day were detected for the proportions of progressively motile (P = 0.02) and rapidly moving (P = 0.03) spermatozoa, and measures of sperm velocity, including path velocity of the smoothed cell path (P = 0.05) and average velocity measured in a straight line from the beginning to the end of the track (P = 0.05). Negative effects of day of semen collection on sperm motility were least pronounced in boars fed Sel-Plex. Experiment 2 was conducted when boars were 17 mo of age, and semen was collected (n = 10 boars/dietary treatment), diluted in commercially available extenders, and stored at 18°C for 9 d. Effects of treatment × day were detected for percentages of motile (P = 0.01) and static (P = 0.01) spermatozoa, amplitude of lateral head displacement (P = 0.02), frequency with which the sperm track crossed the sperm path (P = 0.04), straightness (P = 0.01), and average size of all sperm heads (P = 0.03). In general, sperm cells from boars fed Sel-Plex were better able to maintain motility during liquid storage compared with boars fed sodium selenite. For Exp. 3, semen was collected from boars (n = 6/dietary treatment) at 23 mo of age, and spermatozoa were evaluated at d 1 and 8 after semen collection using in vitro fertilization procedures. There was a tendency for an effect (P = 0.11) of dietary treatment on fertilization rate with Sel-Plex-fed boars having the greatest value (70.7%). The results of this study suggest that there are positive effects of dietary supplementation with Sel-Plex on boar semen characteristics and that organic Se supplementation may help ameliorate the negative effects of semen storage on characteristics of sperm motility.

Effects of organic selenium supplementation on growth performance, carcass measurements, tissue selenium concentrations, characteristics of reproductive organs, and testis gene expression profiles in boars.

The objective was to compare growth and physiological responses in boars fed diets supplemented with organic or inorganic sources of Se. At weaning, crossbred boars (n = 117; 8.3 kg of BW) were placed in nursery pens (3 boars/pen) and assigned within BW blocks to receive on an ad libitum basis 1 of 3 dietary treatments: I) basal diets with no supplemental Se (controls), II) basal diets supplemented with 0.3 mg/kg of organic Se, and, III) basal diets supplemented with 0.3 mg/kg of sodium selenite (13 pens/dietary treatment). Average daily gain (470 g/d), ADFI (896 g/d), and G:F (0.54) were similar among groups. Blood Se concentrations were greater (P < 0.01) for boars consuming organic Se (107.5 ± 4.8 µg/L) or sodium selenite (114.7 ± 4.8 µg/L) compared with controls (28.4 ± 4.8 µg/L). Intact pens of boars (11 pens/dietary treatment) were moved to a grow-finish barn and continued to receive appropriate diets on an ad libitum basis. Average daily gain (1,045 g/d) and ADFI (2,716 g/d) were similar among groups. Gain:feed was affected by treatment (P = 0.02) and was greater (P < 0.06) for boars fed organic Se (0.378 ± 0.004) compared with boars fed sodium selenite (0.368 ± 0.004) or controls (0.363 ± 0.004). Blood Se concentrations were greater (P < 0.01) in grow-finish boars consuming organic Se (198.9 ± 5.5 µg/L) than boars consuming sodium selenite (171.4 ± 5.4 µg/L) or controls (26.7 ± 5.4 µg/L). Treatment did not affect (P > 0.15) HCW, dressing percent, carcass length, LM area, standardized fat-free lean, lean percentage, backfat thickness, visual color, firmness, marbling, or Minolta loin color scores. Selenium supplementation did not affect (P > 0.17) testis or accessory sex gland sizes. Concentrations of Se in loin, liver, kidney, testis, cauda epididymis, and accessory sex glands were greatest (P < 0.01) in boars receiving organic Se, intermediate in boars receiving sodum selenite, and least in control boars. Microarray analysis of testis gene expression did not detect differences (P > 0.05) due to dietary treatment. Testis gene expression of glutathione peroxidase 4, as determined using quantitative PCR, was increased (P < 0.01) in boars fed organic Se compared with those fed sodium selenite. In summary, dietary supplementation of boars with organic Se failed to alter ADG or ADFI but enhanced G:F during grow-finish. More research is needed to discern the mechanism by which organic Se improves feed efficiency in boars.

Dietary supplementation with a source of omega-3 fatty acids increases sperm number and the duration of ejaculation in boars.

Development of nutritional strategies to increase the production of fertile sperm would further enhance the distribution of superior genetic material by AI. The objective was to determine the effects of a dietary source of omega-3 fatty acids in boars on semen characteristics and sexual behavior. Boars were fed daily 2.2 kg of a diet top-dressed with 0.3 kg of corn (controls; n=12) or 0.3 kg of a supplement containing 31% omega-3 fatty acids (n=12) for 16 weeks. Semen was collected weekly and for boars that received the supplement containing omega-3 fatty acids, total sperm per ejaculate averaged 84.3+/-2.3 x 10(9) (mean+/-S.E.M.) during Weeks 0-7, and increased (P=0.02) to 95.6+/-2.3 x 10(9) during Weeks 8-15. Control boars averaged 86.3+/-2.3 x 10(9) sperm per ejaculate during Weeks 0-7 and 86.4+/-2.3 x 10(9) during Weeks 8-15. Other semen characteristics were similar (P>0.1) between groups. Duration of ejaculation was affected by treatment (343.9s for controls and 388.8s for boars fed omega-3 fatty acids; S.E.M.=15.7; P=0.05). In summary, semen characteristics and sexual behavior were altered in boars fed a supplement containing omega-3 fatty acids. Boar semen is typically diluted to create AI doses containing 3 x 10(9) sperm each; therefore, use of the supplement increased the number of potential AI doses by approximately three per ejaculate after the initial 7 week supplementation period.

Growth performance and intestinal morphology responses in early weaned pigs to supplementation of antibiotic-free diets with an organic copper complex and spray-dried plasma protein in sanitary and nonsanitary environments.

The objective of this study was to determine the effects of addition of spray-dried plasma protein (SDPP) and Cu to nonmedicated diets on growth performance and intestinal morphology in weaned pigs reared in sanitary or nonsanitary environments. Weanling pigs (n = 192, 18 +/- 2 d of age, 6.0 +/- 0.2 kg of BW) were assigned to 8 treatments arranged factorially, including 2 dietary levels of SDPP (0 or 6% for the initial 10 d), 2 levels of added dietary Cu (0 or 200 ppm for the entire 35-d experiment), and 2 pen sanitation conditions (sanitized or nonsanitized before pig placement). The nonsanitary pen condition was created by 3 applications of swine manure slurry to all pen surfaces in 1 room and not washing or disinfecting. In an identical adjacent room, sanitary pens were washed and disinfected before weaning. There were 4 pigs per pen, and feed and water were available ad libitum. Growth performance was determined at the end of each diet formulation phase (d 10, 20, and 35 after weaning). On d 10, 1 pig per pen was euthanized, and cross sections of duodenum, jejunum, and ileum were collected for microscopic assessment of mucosal morphology. During the initial postweaning period, SDPP, and Cu supplementation improved ADG and ADFI (P < 0.001). A trend for an interaction of sanitation x dietary SDPP (P = 0.07) was observed for G:F, with a positive response to the supplement in nonsanitary pens but no response in sanitary pens. There were no interactions of SDPP and Cu for any performance variables (P > 0.30). By d 35, there were no main or interaction effects of treatment on ADG or G:F (P > 0.17). Pen sanitation condition produced morphological effects, with shorter villous length and less crypt depth observed in each intestinal segment for pigs reared in the nonsanitary pens (P < 0.05), but these effects must be considered conditional based on the potential confounding influence of separate nursery rooms. In the duodenum, reduced crypt depth with Cu supplementation (P = 0.01) and a tendency for greater villous length with SDPP supplementation (P = 0.09) were observed. In this study, SDPP and Cu supplementation improved pig growth performance during the initial 10-d postweaning. These modifications to nonmedicated diets acted independently with regard to their impacts on postweaning performance and, therefore, could have additive effects.

Life-threatening dystonia-dyskinesias in a child: successful treatment with bilateral pallidal stimulation.

We report a 13-year-old boy who developed severe, refractory dystonia-dyskinesias as an abrupt worsening of a previously nonprogressive movement disorder. The movements became continuous, requiring artificial respiration and continuous sedation in the intensive-care unit. Various drugs and drug combinations failed to achieve control. The child was then treated successfully with bilateral pallidal (GPi) stimulation as shown in the videotape. Four months later and without medication, the boy regained autonomous gait and audible speech; his neurologic condition continues to improve.

On the site of action of naloxone-stimulated cortisol secretion in gilts.

The increase in serum cortisol concentrations following naloxone administration to female pigs was abolished by hypophysial stalk-transection, even though CRH and ACTH stimulated cortisol release in these animals. We suggest that the opioid antagonist enhances cortisol secretion primarily by a central action in pigs.