Effect of diet and nonesterified fatty acid levels on global transcriptomic profiles in circulating peripheral blood mononuclear cells in early lactation dairy cows.

After calving, lipid mobilization caused by increased nutrient demands for lactation leads to elevated circulating concentrations of nonesterified fatty acids (NEFA). Excessive NEFA levels have previously been identified as a major risk factor for postpartum immunosuppression. The aim of this study was to investigate changes in global transcriptomic gene expression of peripheral blood mononuclear cells (PBMC) in dairy cows offered different early lactation diets (high concentrate, n = 7; medium, n = 8; or low, n = 9) and with differing circulating levels of NEFA. Cows were classified as having NEFA concentrations of either <500 µM (low, n = 6), 500 to 750 µM (medium, n = 8) or >750 µM (high, n = 10) at 14 d in milk. Plasma urea concentrations were greater for cows on the high concentrate diet but ß-hydroxybutyrate and glucose concentrations did not differ significantly between either dietary treatments or NEFA groups. Cows with high NEFA weighed more at drying off and suffered greater body condition score loss after calving. The PBMC were isolated at 14 d in milk, and RNA was extracted for RNA sequencing. Differential gene expression was analyzed with DESeq2 with q-value for false discovery rate control followed by Gene Ontology Enrichment. Although there were no differentially expressed genes associated with lactation diet, 304 differentially expressed genes were identified between cows with high and low circulating NEFA, with 118 upregulated and 186 downregulated. Gene Ontology enrichment analysis demonstrated that biological adhesion and immune system process were foremost among various PBMC functions which were altered relating to body defenses and immunity. High NEFA concentrations were associated with inhibited cellular adhesion function by downregulating 20 out of 26 genes (by up to 17-fold) related to this process. Medium NEFA concentrations altered a similar set of functions as high NEFA, but with smaller enrichment scores. Localization and immune system process were most significant, with biological adhesion ranking only eleventh. Our results demonstrated that increased circulating NEFA concentrations, but not diet, were associated with immune system processes in PBMC in early lactation cows. Leukocyte cell-to-cell adhesion was inhibited when the NEFA concentration exceeded 750 µM, which would reduce the efficiency of diapedesis and so contribute to decreased body defense mechanisms and predispose animals to infection.

Anxiety-associated and separation distress-associated behaviours in Angelman syndrome.

BACKGROUND: Anxiety is considered a 'frequent' feature in the clinical criteria for Angelman syndrome; however, the nature and severity of anxiety symptoms have not been well characterised in this population. Anxiety behaviours, especially in response to separation from a preferred caregiver, have been described clinically but have not yet been explored empirically. METHOD: This study used a combination of standardised and clinician-derived survey items to assess the frequency, nature and severity of behaviours associated with anxiety and separation distress in 100 individuals with Angelman syndrome. Family (e.g. income and maternal education) and individual (e.g. age, sex, genetic subtype, sleep difficulties and aggressive behaviours) variables were also gathered to assess possible predictors of higher anxiety levels. Approximately half of the sample was seen in clinic and assessed with standardised measures of development and daily functioning, allowing for an additional exploration of the association between anxiety symptoms and extent of cognitive impairment. RESULTS: Anxiety concerns were reported in 40% of the sample, almost 70% were reported to have a preferred caregiver and over half displayed distress when separated from that caregiver. Individuals with the deletion subtype and individuals who are younger were less likely to have anxiety behaviours. Sleep difficulties and aggressive behaviour consistently significantly predicted total anxiety, the latter suggesting a need for future studies to tease apart differences between anxiety and aggression or anger in this population. CONCLUSIONS: Anxiety concerns, especially separation distress, are common in individuals with Angelman syndrome and represent an area of unmet need for this population.

Mobile technology use and skills among individuals with fragile X syndrome: implications for healthcare decision making.

BACKGROUND: Little is known about how individuals with fragile X syndrome (FXS) and their families use technology in daily life and what skills individuals with FXS can perform when using mobile technologies. METHODS: Using a mixed-methods design, including an online survey of parents (n = 198) and a skills assessment of individuals with FXS (n = 6), we examined the experiences and abilities of individuals with FXS for engaging with mobile technology. RESULTS: Parents reported that individuals with FXS often used technology in their daily lives, with variations based on age of child, sex, autism status, depression, and overall ability. Parents frequently sought and shared FXS-related information online. Assessment data revealed that individuals with FXS demonstrated proficiency in interacting with technology. CONCLUSIONS: Mobile technology is a tool that can be used in FXS to build skills and increase independence rather than simply for recreational purposes. Implications for using mobile technology to enhance healthcare decision making are discussed.

Effects of dry period length and concentrate protein content in late lactation on body condition score change and subsequent lactation performance of thin high genetic merit dairy cows.

Improving body condition score of thin cows in late lactation is necessary, because cows that are thin at drying off exhibit decreased fertility postpartum and are at increased risk of disease and of being culled in the subsequent lactation. Offering a diet low in crude protein (CP) content in late lactation may help to improve body condition score (BCS) at drying off, whereas imposing an extended dry period (EDP) has been advocated as another way to increase BCS at calving. To test these hypotheses, 65 thin cows (mean BCS 2.25 at 14 wk precalving) were managed on 1 of 3 treatments between 13 and 9 wk prepartum: normal protein control {NP; grass silage + 5 kg/d of a normal protein concentrate [228 g of CP/kg of dry matter (DM)]}, low protein [LP; grass silage + 5 kg/d of a low-protein concentrate (153 g of CP/kg of DM)], or EDP (cows dried off at 13 wk precalving and offered a grass silage-only diet). Both NP and LP cows were dried off at wk 8 prepartum, after which all cows were offered a grass silage-only diet until calving. After calving, all cows were offered a common diet (supplying 11.1 kg of concentrate DM/cow per day) for 19 wk. Between 13 and 9 wk prepartum, LP cows had lower DM intake, milk yield, and body weight than NP cows. Whereas EDP cows had lower serum ß-hydroxybutyrate and fatty acid concentrations than those of NP cows, BCS at wk 9 prepartum did not differ between treatments. Cows on the LP treatment continued to have lower DMI and BW than those of NP and EDP cows between 8 wk prepartum and calving, but only EDP cows had a higher BCS at calving. Treatment did not affect calving difficulty score or calf birth weight. Although all cows were offered a common diet postpartum, cows on the LP treatment had lower DM intake and milk fat + plus protein yield than cows on any other treatment during the 19-wk period postpartum, but we found no differences in any postpartum indicator of body tissue reserves. The treatments imposed from wk 13 to 9 prepartum had no effect on any fertility or health parameters examined postpartum. Extending the dry period for thin cows improved their BCS at calving but did not allow these cows to achieve the target BCS of 2.75, and we found no beneficial effects of this treatment on cow performance postpartum. Offering a lower-protein diet to thin cows in late lactation did not improve BCS at calving above that of cows on a normal protein diet, but had unexplained long-term negative effects on cow performance.

Intergenerational effects of racism on amygdala and hippocampus resting state functional connectivity.

Racism is an insidious problem with far-reaching effects on the lives of Black, Indigenous, and People of Color (BIPOC). The pervasive negative impact of racism on mental health is well documented. However, less is known about the potential downstream impacts of maternal experiences of racism on offspring neurodevelopment. This study sought to examine evidence for a biological pathway of intergenerational transmission of racism-related trauma. This study examined the effects of self-reported maternal experiences of racism on resting state functional connectivity (rsFC) in n = 25 neonates (13 female, 12 male) birthed by BIPOC mothers. Amygdala and hippocampus are brain regions involved in fear, memory, and anxiety, and are central nodes in brain networks associated with trauma-related change. We used average scores on the Experiences of Racism Scale as a continuous, voxel-wise regressor in seed-based, whole-brain connectivity analysis of anatomically defined amygdala and hippocampus seed regions of interest. All analyses controlled for infant sex and gestational age at the 2-week scanning session. More maternal racism-related experiences were associated with (1) stronger right amygdala rsFC with visual cortex and thalamus; and (2) stronger hippocampus rsFC with visual cortex and a temporo-parietal network, in neonates. The results of this research have implications for understanding how maternal experiences of racism may alter neurodevelopment, and for related social policy.

Transcriptional response to GAA deficiency (Pompe disease) in infantile-onset patients.

Pompe disease is a genetic disorder resulting from a deficiency of lysosomal acid alpha-glucosidase (GAA) that manifests as a clinical spectrum with regard to symptom severity and rate of progression. In this study, we used microarrays to examine gene expression from the muscle of two cohorts of infantile-onset Pompe patients to identify transcriptional differences that may contribute to the disease phenotype. We found strong similarities among the gene expression profiles generated from biceps and quadriceps, and identified a number of signaling pathways altered in both cohorts. We also found that infantile-onset Pompe patient muscle had a gene expression pattern characteristic of immature or regenerating muscle, and exhibited many transcriptional markers of inflammation, despite having few overt signs of inflammatory infiltrate. Further, we identified genes exhibiting correlation between expression at baseline and response to therapy. This combined dataset can serve as a foundation for biological discovery and biomarker development to improve the treatment of Pompe disease.

Presentation and management of school bullying and the impact of anti-bullying strategies for pupils: a self-report survey in London schools.

OBJECTIVES: To investigate differences in the presentation and management of bullying in two London boroughs, one with high levels of deprivation and the other with low levels of deprivation; and to explore pupils' views on successful approaches to tackle bullying. STUDY DESIGN: Problems associated with bullying in childhood can manifest with mental disorders, depression and poorer perceived health in adulthood. However, despite the recent development of anti-bullying policies within schools, bullying remains a significant issue for many pupils. An original self-report survey was undertaken to investigate this important area. METHOD: A web-based questionnaire was undertaken with pupils from year 8 (aged 12-13 years) at four secondary schools in London. The study was approved by King's College London Research Ethics Committee. RESULTS: In total, 304 pupils completed the questionnaire, 55% of whom were affected by bullying at their school. Deprivation had no impact on the prevalence of pupils involved in a bullying culture. School membership is the most important independent factor for determining bullying. CONCLUSIONS: Policy could be aimed at changing pupils' views on dealing with bullying, and it is vital that policies and practices around bullying are active and alive and do not become 'file fodder'.

An evaluation of the effect of altering nutrition and nutritional strategies in early lactation on reproductive performance and estrous behavior of high-yielding Holstein-Friesian dairy cows.

Reproductive performance in the high-yielding dairy cow has severely decreased in the last 40 yr. The aim of this study was to compare the effectiveness of 4 nutritional strategies in improving the reproductive performance of high-yielding dairy cows. It was hypothesized that offering cows a high-starch ration in early lactation would enhance the onset of luteal activity, and that decreasing the severity of negative energy balance in the early postcalving period would improve reproductive parameters. Nutritional regimens aimed at improving fertility were applied to 96 Holstein-Friesian dairy animals. Upon calving, animals were allocated in a balanced manner to one of 4 dietary treatments. Primiparous animals were balanced according to live weight, body condition score and calving date. Multiparous animals were balanced according to parity, previous lactation milk yield, liveweight, body condition score and calving date. Treatment 1 was based on an industry best practice diet (control) to contain 170 g of crude protein/kg of dry matter. Treatment 2 was an individual cow feeding strategy, whereby the energy balance (EB) of individual animals was managed so as to achieve a predetermined target daily EB profile (±10 MJ/d). Treatment 3 was a high-starch/high-fat combination treatment, whereby an insulinogenic (high-starch) diet was offered in early lactation to encourage cyclicity and followed by a lipogenic (low-starch, high-fat) diet to promote embryo development. Treatment 4 was a low-protein diet, containing 140 g of crude protein/kg of dry matter, supplemented with protected methionine at an inclusion level of 40 g per animal per day. The nutritional strategies implemented in this study had no statistically significant effects on cow fertility measures, which included the onset of luteal activity, conception rate, in-calf rate, and the incidence of atypical cycles. The individual cow feeding strategy improved EB in early lactation but had no benefit on conception rate to first insemination. However, conception rate to second insemination, 100-d pregnancy rate (from the commencement of breeding), and overall pregnancy rate tended to be higher in this group. The high-starch/high-fat treatment tended to decrease the proportion of delayed ovulations and increase the proportion of animals cycling by d 50 postcalving. Animals that failed to conceive to first insemination had a significantly longer luteal phase in the first cycle postpartum and a longer inter-ovulatory interval in the second cycle postpartum. With regards to estrous behavior, results indicate that as the size of the sexually active group increased, the intensity of estrus and the expression of mounting or attempting to mount another cow also increased. Furthermore, cows that became pregnant displayed more intense estrous behavior than cows that failed to become pregnant.

Effect of precalving and postcalving dietary energy level on performance and blood metabolite concentrations of dairy cows throughout lactation.

The effects of the level of energy intake (high E and low E) offered before and after calving on body condition score at calving, production performance, and energy status in the first 250 d of lactation were evaluated in a 2 × 2 factorial design experiment involving 80 Holstein-Friesian dairy animals (40 primiparous and 40 multiparous). From d 80 until d 21 precalving, primiparous animals were offered either high or low pasture allowances. Thereafter, these animals were housed and had ad libitum access to a high energy density diet (high E) or restricted access [6 kg of dry matter (DM) per d] to a low energy density diet (low E), respectively, until calving. From d 100 until d 42 precalving, multiparous animals were offered either ad libitum or restricted (10 kg of DM/d) access to a late lactation diet, and thereafter, had ad libitum access to a high E diet or restricted access (7 kg of DM complete diet/d) to a low E diet, respectively, until calving. The forage to concentrate (F:C) ratios (DM basis) of these high E and low E diets [d 42 (d 21 in primiparous animals) until calving] were 64:36 and 83:17, respectively. Cows offered high E and low E precalving diets were allocated to either a high E or low E postcalving diet [F:C ratio (DM basis) of 30:70 and 70:30, respectively] and remained on these diets until d 250 of lactation. Multiparous animals offered a high E diet precalving had a significantly higher body condition score at calving than those offered the low E diet precalving. This effect was not evident in primiparous animals. Precalving diet had no significant effect on plasma nonesterified fatty acid concentrations during the last 3 wk precalving in primi- or multiparous animals. Primiparous animals offered a high E diet precalving had significantly higher postcalving plasma concentrations of nonesterified fatty acid, suggesting greater mobilization of body reserves. Primi- and multiparous animals offered a high E diet postcalving had a significantly higher dry matter intake, milk yield, and energy status postcalving compared with animals offered a low E diet postcalving. Milk yields of primiparous animals offered high E and low E diets postcalving were 29.7 and 24.8 kg/d, respectively, and milk yield of multiparous animals offered high E and low E diets postcalving were 33.5 and 28.2 kg/d, respectively. It is concluded that altering body condition score during the dry period is difficult but that specific dietary regimens applied precalving can have a significant influence on postcalving production and energy-related parameters.

Effect of dietary restriction and compensatory growth on performance, carcass characteristics, and metabolic hormone concentrations in Angus and Belgian Blue steers.

Compensatory growth (CG) is the ability of an animal to undergo accelerated growth after a period of restricted feeding. However, there is a dearth of information in relation to the effect of genotype on CG response, thus the objective of this study was to evaluate CG response in two contrasting breed types, namely Aberdeen Angus (AN) and Belgian Blue (BB). Crossbred AN × Holstein-Friesian or BB × Holstein-Friesian steers were assigned to one of two treatment groups in a two (genotypes) × two (diets) factorial design. For 99 days, one group (11 AN and 12 BB) was offered a high energy control diet (H-H) whereas the second group (11 AN and 12 BB) was offered an energy restricted diet (L-H). At the end of the differential feeding period (99 days), both groups of animals were then offered a high energy control diet for a further 200 days. All animals were then slaughtered on day-299 of the study. During feed restriction, L-H had lower DM intake (DMI), had greater feed conversion ratio (FCR) and lower plasma concentrations of insulin, IGF-1, leptin, glucose, urea, betahydroxybutyrate and smaller M. longissimus thoracis or lumborum muscle and fat depths compared to H-H steers. During realimentation, there was no difference in DMI between diets; however, L-H had greater live weight gain compared to H-H steers. Overall, H-H consumed greater quantities on a DM basis, however, had a higher FCR compared to L-H steers. By the end of the realimentation period, there was no difference in plasma metabolite or hormone concentrations, linear body measurements, ultrasonically scanned fat depths, carcass conformation, dressing percentage or fat class between H-H and L-H steers. At slaughter, carcass weights were affected by diet with greater values for H-H compared to L-H steers. Genotype affected measures associated with body composition including pelvic width and both muscle and fat depths (P < 0.05). Overall, L-H had a CG (or recovery) index of 0.52 and did not make up for the loss of gains during the differential feeding period; however, M. longissimus thoracis et lumborum, a tissue of high economic value, recovered completely making it a target of interest for further investigation.

Effect of dietary protein content on the fertility of dairy cows during early and mid lactation.

Ninety autumn-calving Holstein dairy cows (45 primiparous and 45 multiparous; mean parity, 3.1) were allocated to 1 of 3 treatments; 173, 144, or 114 g of crude protein (CP)/kg of dry matter (DM) from calving until d 150 of lactation. On d 151 of lactation, half the animals receiving 114 g of CP/kg of DM went onto 144 g of CP/kg of DM, half of the animals receiving 144 g of CP/kg of DM went onto 173 g of CP/kg of DM, and half of the animals receiving 173 g of CP/kg of DM went onto 144 g of CP/kg of DM, with the remaining animals staying on their original treatments. This resulted in 6 treatments in mid to late lactation: 114/114; 144/144; 173/173; 114/144; 144/173; and 173/144 g of CP/kg of DM. Overall, 95.3% of cows intended for breeding conceived during a 6-mo breeding period. The average pregnancy rates to first service and first plus second service were 30.9% [standard error of the difference (SED), 0.05] and 56.7% (SED, 0.05) respectively. The average 100 d in-calf rate from the start of the breeding period was 70.5%, and at least one abnormal progesterone profile was observed in 62% of animals. An increase in dietary protein content decreased the requirement for treatment of metritis. There was no effect of dietary protein content on any of the reproductive or progesterone measures; for example, days to conception, calving interval, 100 d in-calf rate (from commencement of breeding), days to onset of luteal activity, average luteal phase, average interovulatory interval, or average interluteal interval. An increase in dietary protein content decreased the average daily energy balance. A more positive energy balance was associated with an increased requirement for the treatment of metritis in the current study. Cumulative energy balance was positively associated with conception. There was no effect of the concentration of plasma urea on any of the reproductive variables; however, the concentration of serum leptin was favorably associated with the time to progesterone increase above 3 ng/mL, which has been deemed essential for embryo survival. Additionally, the average peak concentration of progesterone and the duration of the average luteal phase were favorably associated with the interval from calving to conception. The latter relationships emphasize the importance of progesterone in achieving and maintaining pregnancy.

Effect of dietary protein content on animal production and blood metabolites of dairy cows during lactation.

Ninety autumn-calving Holstein dairy cows [45 primiparous and 45 multiparous (mean parity, 3.1)] were allocated to 1 of 3 dietary crude protein (CP) concentrations: 173, 144, or 114 g of CP/kg of DM, from calving until d 150 of lactation. On d 151, half of the animals in each treatment were allocated an alternative dietary protein concentration. Half of the animals receiving 114 g of CP/kg of DM went onto 144 g of CP/kg of DM; half of the animals receiving 144 g of CP/kg of DM went onto 173 g of CP/kg of DM; and half of the animals receiving 173 g of CP/kg of DM went onto 144 g of CP/kg of DM, with the remaining animals staying on their original treatment. This resulted in 6 treatments in the mid to late lactation period: 114/114, 144/144, 173/173, 114/144, 144/173, and 173/144 g of CP/kg of DM. An increase in dietary CP concentration significantly increased milk, fat, and protein yield in early lactation (d 1 to 150). Dry matter intake was also increased with increased dietary protein concentration; however, this was not significant between 144 and 173 g of CP/kg of DM. Increased dietary CP significantly increased plasma urea, albumin, and total protein concentrations but had no significant effect on NEFA, leptin, or IGF-1 concentrations. Decreasing the dietary CP concentration in mid-late lactation (d 151 to 305) from 173 to 144 g/kg of DM had no significant effect on milk yield, dry matter intake, or milk fat and protein yield, compared with animals that remained on 173 g of CP/kg of DM throughout lactation. Increasing dietary CP concentration from 144 to 173 g/kg of DM significantly increased dry matter intake compared with animals that remained on the 144 g of CP/kg of DM throughout lactation. There were no significant dietary treatment effects on live weight or body condition score change throughout the experiment. Results of this study indicate that high protein diets (up to 173 g of CP/kg of DM) improved feed intake and animal performance in early lactation (up to d 150), but thereafter, protein concentration can be reduced to 144 g of CP/kg of DM with no detrimental effects on animal performance.

Effect of dietary protein content on estrous behavior of dairy cows during early and mid lactation.

One of the main contributing factors to the decline in fertility in contemporary dairy farming is the inability to detect cows in estrus. In the current study, 90 Holstein dairy cows [45 primiparous and 45 multiparous (mean parity of 3.1)] were allocated to 1 of 3 treatments at calving; 173, 144, or 114 g of crude protein/kg of dry matter. Estrous behavior was recorded for one 30-min period every 12 h from calving until all animals reached 140 d postpartum. Behavioral activities were recorded according to a scoring system developed by Van Eerdenburg et al. (1996), with 9 key estrous behavioral activities each allocated a given number of points. If the total score allocated was greater than or equal to 50 points during a single or consecutive observational periods, then the animal was deemed to be in estrus. A total of 238 estrous cycles scored 50 points or above on the Van Eerdenburg et al. (1996) scale in this experiment, with 51.7% of these cycles being characterized as standing immobile on mounting. There were no direct effects of dietary protein content on estrous behavior; however, 3 significant stage of lactation x protein treatment interactions occurred for the behavioral activities (mucous discharge, chin resting, and mounting the head side of another cow), but no consistent trends were apparent from the predicted means. There was a significant influence of parity on the frequency of mounting the head side of another cow and total number of behavior activities displayed per estrous cycle. In both cases multiparous animals displayed fewer behavioral activities than primiparous animals. An increase in the size of the sexually active group (animals in estrus at the same time, up to 5) significantly increased the expression of mounting or attempting to mount another cow, the number of cycles in which standing immobile on being mounted was observed, the total estrous score and the proportion of cyclic animals that were diagnosed as being in estrus. The most frequent behavioral activity displayed was chin resting (89.5% of cycles), and the most reliable behavior was standing immobile on mounting (when expressed 96.4% of cows were in estrus). The most dependable (function of reliability and frequency displayed) sign of estrus was mounting or attempting to mount another cow. This behavior was expressed in 83% of cycles, and when expressed, 89% of animals were in estrus.

Immunological effects of altering the concentrate inclusion level in a grass silage-based diet for early lactation Holstein Friesian cows.

Concentrate inclusion levels in dairy cow diets are often adjusted so that the milk yield responses remain economic. While changes in concentrate level on performance is well known, their impact on other biological parameters, including immune function, is less well understood. The objective of this study was to evaluate the effect of concentrate inclusion level in a grass silage-based mixed ration on immune function. Following calving 63 (45 multiparous and 18 primiparous) Holstein Friesian dairy cows were allocated to one of three isonitrogenous diets for the first 70 days of lactation. Diets comprised of a mixture of concentrates and grass silage, with concentrates comprising either a low (30%, LC), medium (50%, MC) or high (70%, HC) proportion of the diet on a dry matter (DM) basis. Daily DM intakes, milk yields and BW were recorded, along with weekly body condition score, milk composition and vaginal mucus scores. Blood biochemistry was measured using a chemistry analyzer, neutrophil phagocytic and oxidative burst assessed using commercial kits and flow cytometry, and interferon-γ production evaluated by ELISA after whole blood stimulation. Over the study period cows on HC had a higher total DM intake, milk yield, fat yield, protein yield, fat+protein yield, protein content, mean BW and mean daily energy balance, and a lower BW loss than cows on MC, whose respective values were higher than cows on LC. Cows on HC and MC had a lower serum non-esterified fatty acid concentration than cows on LC (0.37, 0.37 and 0.50 mmol/l, respectively, P=0.005, SED=0.032), while cows on HC had a lower serum ß-hydroxybutyrate concentration than cows on MC and LC (0.42, 0.55 and 0.55 mmol/l, respectively, P=0.002, SED=0.03). Concentrate inclusion level had no effect on vaginal mucus scores. At week 3 postpartum, cows on HC tended to have a higher percentage of oxidative burst positive neutrophils than cows on LC (43.2% and 35.3%, respectively, P=0.078, SED=3.11), although at all other times concentrate inclusion level in the total mixed ration had no effect on neutrophil phagocytic or oxidative burst characteristics, or on interferon-γ production by pokeweed mitogen stimulated whole blood culture. This study demonstrates that for high yielding Holstein Friesian cows managed on a grass silage-based diet, concentrate inclusion levels in early lactation affects performance but has no effect on neutrophil or lymphocyte immune parameters.

Biochemical and pharmacological characterization of different recombinant acid alpha-glucosidase preparations evaluated for the treatment of Pompe disease.

Pompe disease results in the accumulation of lysosomal glycogen in multiple tissues due to a deficiency of acid alpha-glucosidase (GAA). Enzyme replacement therapy for Pompe disease was recently approved in Europe, the U.S., Canada, and Japan using a recombinant human GAA (Myozyme, alglucosidase alfa) produced in CHO cells (CHO-GAA). During the development of alglucosidase alfa, we examined the in vitro and in vivo properties of CHO cell-derived rhGAA, an rhGAA purified from the milk of transgenic rabbits, as well as an experimental version of rhGAA containing additional mannose-6-phosphate intended to facilitate muscle targeting. Biochemical analyses identified differences in rhGAA N-termini, glycosylation types and binding properties to several carbohydrate receptors. In a mouse model of Pompe disease, glycogen was more efficiently removed from the heart than from skeletal muscle for all enzymes, and overall, the CHO cell-derived rhGAA reduced glycogen to a greater extent than that observed with the other enzymes. The results of these preclinical studies, combined with biochemical characterization data for the three molecules described within, led to the selection of the CHO-GAA for clinical development and registration as the first approved therapy for Pompe disease.

Periprandial changes in metabolite and metabolic hormone concentrations in high-genetic-merit dairy heifers and their relationship to energy balance in early lactation.

Sixteen high-genetic-merit Holstein-Friesian heifers were offered a complete diet of grass silage, maize silage, and concentrates ad libitum through mo 1 to 3 postpartum. Open-circuit calorimetry and a 6-d digestibility balance were performed on each heifer at the end of each month, and energy balance (EB) was calculated. After each digestibility balance, heifers were blood sampled hourly from 1 h before to 9 h after feeding. Prefeed plasma and 3-h composites of postfeed plasmas were analyzed for selected metabolites and hormones. Levels of nonesterified fatty acids (NEFA) decreased, whereas those of beta-hydroxybutyrate (BHBA) increased after feeding in each month. Urea levels increased after feeding, whereas glucose levels decreased in each month. Insulin increased after feeding but with reducing significance as lactation progressed. Insulin was always lower before feeding, and mean insulin increased from mo 1 to mo 3. Levels of insulin-like growth factor-1 (IGF-I) increased across mo 1 to 3, but were unaffected by feeding except in mo 2, whereas leptin levels varied significantly on each sampling occasion, but showed no increase between mo 1 and mo 3. Multiple regression of all data showed no significant correlation between EB and either BHBA or NEFA levels. However, EB was negatively correlated with leptin levels (r = -0.632), which were themselves positively associated with IGF-I (r = 0.498) and glucose (r = 0.565). A relationship was established between overall mean EB and prefeed leptin, insulin, and urea (R(2) = 0.63) in the first 3 mo of lactation. A potentially useful relationship was also established between EB and prefeed concentrations of leptin, IGF-I, urea, and glucose (R(2) = 0.80) only in mo 1 of lactation.

Estimation of the maintenance energy requirements, methane emissions and nitrogen utilization efficiency of two suckler cow genotypes.

Seventeen non-lactating dairy-bred suckler cows (LF; Limousin×Holstein-Friesian) and 17 non-lactating beef composite breed suckler cows (ST; Stabiliser) were used to study enteric methane emissions and energy and nitrogen (N) utilization from grass silage diets. Cows were housed in cubicle accommodation for 17 days, and then moved to individual tie-stalls for an 8-day digestibility balance including a 2-day adaption followed by immediate transfer to an indirect, open-circuit, respiration calorimeters for 3 days with gaseous exchange recorded over the last two of these days. Grass silage was offered ad libitum once daily at 0900 h throughout the study. There were no significant differences (P>0.05) between the genotypes for energy intakes, energy outputs or energy use efficiency, or for methane emission rates (methane emissions per unit of dry matter intake or energy intake), or for N metabolism characteristics (N intake or N output in faeces or urine). Accordingly, the data for both cow genotypes were pooled and used to develop relationships between inputs and outputs. Regression of energy retention against ME intake (r 2=0.52; P<0.001) indicated values for net energy requirements for maintenance of 0.386, 0.392 and 0.375 MJ/kg0.75 for LF+ST, LF and ST respectively. Methane energy output was 0.066 of gross energy intake when the intercept was omitted from the linear equation (r 2=0.59; P<0.001). There were positive linear relationships between N intake and N outputs in manure, and manure N accounted for 0.923 of the N intake. The present results provide approaches to predict maintenance energy requirement, methane emission and manure N output for suckler cows and further information is required to evaluate their application in a wide range of suckler production systems.

Assessment of Medicaid managed behavioral health care for persons with serious mental illness.

OBJECTIVES: This five-site study compared Medicaid managed behavioral health programs and fee-for-service programs on use and quality of services, satisfaction, and symptoms and functioning of adults with serious mental illness. METHODS: Adults with serious mental illness in managed care programs (N=958) and fee-for-service programs (N=1,011) in five states were interviewed after the implementation of managed care and six months later. After a multiple regression to standardize the groups for case mix differences, a meta-analysis using a random-effects model was conducted, and bioequivalence methods were used to determine whether differences were significant for clinical or policy purposes. RESULTS: A significantly smaller proportion of the managed care group received inpatient care (5.7 percent compared with 11.5 percent). The managed care group received significantly more hours of primary care (4.9 compared with 4.5 hours) and was significantly less healthy. However, none of these differences exceed the bioequivalence criterion of 5 percent. Managed care and fee for service were "not different but not equivalent" on 20 of 34 dependent variables. Cochrane's Q statistic, which measured intersite consistency, was significant for 20 variables. CONCLUSIONS: Managed care and fee-for-service Medicaid programs did not differ on most measures; however, a lack of sufficient power was evident for many measures. Full endorsement of managed care for vulnerable populations will require further research that assumes low penetration rates and intersite variability.