Effects of abomasal infusions of fatty acids and 1-carbon donors on apparent fatty acid digestibility and incorporation into milk fat in cows.

Our primary objective was to determine the effects of the abomasal infusion of 16-carbon (16C) and 22-carbon (22C) fatty acids (FA) on apparent FA digestibility, plasma FA concentrations, and their incorporation into milk fat in cows. Our secondary objective was to study the effects of 1-carbon donors choline and l-serine on these variables. Five rumen-cannulated Holstein cows (214 ± 4.9 d in milk; 3.2 ± 1.1 parity) were enrolled in a 5 × 5 Latin square experiment with experimental periods lasting 6 d. Abomasal infusates consisted of (1) palmitic acid (PA; 98% 16:0 of total fat), (2) PA + choline chloride (PA+CC; 50 g/d of choline chloride), (3) PA + l-serine (PA+S; 170 g/d of l-serine), (4) behenic acid (BA; 92% 22:0 of total fat), and (5) docosahexaenoic acid algal oil (DHA; 47.5% DHA of total fat). Emulsions were formulated to provide 301 g/d of total FA and were balanced to provide a minimum of 40 and 19 g/d of 16:0 and glycerol, respectively, to match the content found in the infused algal oil. Apparent digestibility of FA was highest in DHA, intermediate in PA, and lowest in BA. Digestibility of 16C FA was lowest in BA and highest in PA. The digestibility of 22C FA was highest in DHA relative to BA (99 vs. 58%), whereas 1-carbon donors had no effect on 22C FA digestibility. Plasma 16C FA concentrations were greatest with PA treatment, and 22C FA concentrations were ~3-fold greater in DHA-treated cows relative to all other treatments. Milk fat 16:0 content was highest in PA relative to BA and DHA (e.g., 37 vs. 27% in PA and DHA), whereas the milk yield of 16:0 was higher in PA relative to DHA (i.e., 454 vs. 235 g/d). Similarly, milk 22:0 content and yield were ~10-fold higher in BA relative to all other treatments, whereas DHA treatment resulted in higher content and yield of 22:6 in milk fat relative to all other treatments (41- and 38-fold higher, respectively). Consequently, the content of FA >16C (i.e., preformed) was higher in milk fat from cows infused with BA and DHA relative to PA. De novo FA content in milk did not differ between PA, PA+CC, and PA+S (~16% of milk fat) but was higher in BA and DHA treatments (19 and 21%, respectively). We conclude that FA carbon chain length and degree of saturation affected FA digestibility and availability for absorption as well as their incorporation into milk fat. The abomasal infusion of choline chloride and l-serine did not modify these variables relative to infusing palmitic acid alone.

Effects of dietary deoiled soy lecithin supplementation on milk production and fatty acid digestibility in Holstein dairy cows.

Deoiled soy lecithin is a feed additive enriched in phospholipids. Our study evaluated the effects of dietary deoiled soy lecithin supplementation on (1) milk production and composition, (2) plasma and milk fatty acid (FA) content and yield, and (3) apparent FA digestibility and absorption in lactating dairy cows fed fractionated palm fat. In a split-plot Latin square design, 16 Holstein cows (160 ± 7 days in milk; 3.6 ± 1.2 parity) were randomly allocated to a main plot receiving a corn silage and alfalfa haylage-based diet with palm fat containing either moderate (MPA) or high palmitic acid (HPA) content at 1.75% of ration dry matter (72 or 99% palmitic acid, respectively; n = 8/palm fat diet). On each palm fat diet, deoiled soy lecithin was top-dressed at 0, 0.12, 0.24, or 0.36% of ration dry matter in a replicated 4 × 4 Latin square design. Following a 14-d covariate period, lecithin supplementation spanned 14 d, with milk and blood collected during the final 3 d. Milk composition and pooled plasma markers were measured. The statistical model included the fixed effects of palm fat type, lecithin dose, period, and the interaction between palm fat type and lecithin dose. The random effect of cow nested within palm fat group was also included. Lecithin linearly decreased dry matter intake. In cows fed HPA, lecithin feeding reduced milk fat content and tended to decrease milk fat yield. Although no changes in milk yield were observed, a quadratic reduction in 3.5% fat-corrected milk was observed with increasing lecithin dose. Lecithin linearly increased energy-corrected milk efficiency in cows fed MPA. Lecithin supplementation also decreased milk urea nitrogen, relative to unsupplemented cows. The proportion of 16-carbon FA in milk fat decreased linearly with lecithin dose, whereas 18-carbon FA increased linearly. Lecithin reduced de novo FA (<16-carbon) content and tended to increase preformed FA (>16-carbon) content in a linear manner. Compared with MPA, HPA diets reduced apparent total and 16-carbon FA digestibility and absorption. Deoiled soy lecithin feeding did not modify FA digestibility or absorption. Our observations suggest that soy lecithin feeding modifies rumen digestion to reduce dry matter intake and change milk composition.

Somatotropin increases plasma ceramide in relation to enhanced milk yield in cows.

Recombinant bovine somatotropin (rBST) changes metabolism to spare glucose for milk synthesis in cows. Ceramides inhibit insulin responsiveness in bovine adipocytes and are associated with insulin resistance and milk production in cows. The mechanisms by which rBST supports lactation may involve ceramide. Eight multiparous lactating Holstein cows were enrolled in a 2 × 2 replicated Latin square design with 14-d periods. Cows received a single rBST injection (Posilac; Elanco Animal Health, Indianapolis, IN; 0.062 mg/kg BW) or no injection (CON). An epinephrine challenge, insulin tolerance test, and liver biopsy were performed. Somatotropin enhanced the conversion of feed nutrients into milk components and increased plasma free fatty acid (FFA) concentrations (P < 0.01). Area-under-the-curves for FFA in response to epinephrine and insulin were greater in rBST-treated cows. In response to insulin, glucose concentrations (20- and 30-min post-challenge) and insulin area-under-the-curve were higher with rBST treatment (P < 0.05, <0.10, and <0.01), suggesting insulin resistance. Somatotropin modified the plasma lipidome. For example, rBST decreased plasma di- and triacylglycerol levels (eg, DG-50:1 and TG-18:0/16:0/16:1), phosphatidylcholines and sphingomyelins (P < 0.05). Somatotropin increased plasma total and very-long-chain (C22:0-, C24:0-, C26:0-) ceramide concentrations (P < 0.01). Liver ceramide concentrations were not modified. Plasma ceramides were positively correlated with circulating FFA (r ~ 0.57; P < 0.05) and milk yield (r ~ 0.63; P < 0.05). We conclude that rBST administration modifies the bovine lipidome and increases plasma ceramide concentrations in association with increased milk production in cows.

Effects of abomasal infusions of fatty acids and one-carbon donors on hepatic ceramide and phosphatidylcholine in lactating Holstein dairy cows.

Our objectives were to (1) determine whether the abomasal infusion of behenic acid (C22:0) elevated hepatic ceramide relative to palmitic acid (C16:0) or docosahexaenoic acid (C22:6n-3) infusion; (2) assess whether the abomasal infusion of choline chloride or l-serine elevated hepatic phosphatidylcholine (PC) in cows abomasally infused with C16:0; and (3) characterize the PC lipidome in cows abomasally infused with C22:6n-3, relative to C16:0 or C22:0 infusion. In a 5 × 5 Latin square design, 5 rumen-cannulated Holstein cows (214 ± 4.9 DIM; 3.2 ± 1.1 parity) were enrolled in a study with 6-d periods. Abomasal infusates consisted of (1) palmitic acid (PA; 98% C16:0); (2) PA + choline chloride (PA+C; 50 g/d choline chloride); (3) PA + l-serine (PA+S; 170 g/d l-serine); (4) behenic acid (BA; 92% C22:0); and (5) an algal oil rich in docosahexaenoic acid (DHA; 44% C22:6n-3). Emulsion infusates provided 301 g/d of total fatty acids containing a minimum of 40 g/d of C16:0. Cows were fed a corn silage-based diet. Milk was collected on d -2, -1, 5, and 6. Blood was collected and liver biopsied on d 6 of each period. Although we did not detect differences in milk yield, milk fat yield and content were lower in cows infused with DHA relative to PA. Plasma triacylglycerol concentrations were lower with DHA treatment relative to PA or BA. Cows infused with DHA had lower plasma insulin concentrations relative to cows infused with PA only. For objective 1, hepatic ceramide-d18:2/22:0 was highest in cows infused with BA relative to other treatments. For objective 2, plasma free choline concentrations were greater in PA+C cows relative to PA; however, we did not observe this effect with PA+S. Plasma total PC concentrations were similar for all treatments. Regarding the hepatic lipidome, a total of 18 hepatic PC were higher (e.g., PC-16:1/18:2) and 25 PC were lower (e.g., PC-16:0/22:6) with PA+C infusion relative to PA. In addition, 17 PC were higher (e.g., PC-20:3/22:5) and 21 PC were lower (e.g., PC-18:0/22:6) with PA+S infusion relative to PA. For objective 3, hepatic concentrations of many individual saturated PC (e.g., PC-18:0/15:0) were lower with DHA relative to other treatments. Hepatic concentrations of highly unsaturated PC with very-long-chain fatty acids (e.g., PC-14:0/22:6) were higher in DHA-infused cows relative to PA, PA+C, PA+S, or BA. The abomasal infusion of emulsions containing palmitic acid, palmitic acid with choline chloride or serine, behenic acid, or docosahexaenoic acid influence the hepatic ceramide and PC profiles of lactating cows.

Circulating low-density lipoprotein ceramide concentrations increase in Holstein dairy cows transitioning from gestation to lactation.

Low-density lipoprotein (LDL) ceramide causes insulin resistance in obese diabetic nonruminants. Because previous work suggests that liver-derived ceramide may impair insulin action in postpartum cows, our objectives were to characterize peripartal changes in lipoprotein ceramides. We further studied the effects of prepartum adiposity on lipoprotein ceramide levels. Twenty-eight pregnant Holstein cows (parity = 3.65 ± 1.62) with lean (body condition score, BCS = 2.97 ± 0.16; body weight, BW = 630 ± 55.2 kg; n = 15) or overweight (BCS = 3.93 ± 0.27; BW = 766 ± 46.1 kg; n = 13) body condition 28 d before expected parturition were evaluated. Sampling occurred on d -20.5 ± 1.74, -13.8 ± 1.71, -7.84 ± 4.07, -6.71 ± 1.00, -3.92 ± 0.64, and -1.28 ± 0.61 (before parturition); daily until d 8 postpartum; and on d 10, 12, 14, 21, and 28. Adipose tissue and liver were biopsied on d -7.84 ± 4.07 and 10. Postpartum insulin sensitivity was assessed using the hyperinsulinemic-euglycemic clamp. Lipoprotein fractions were isolated using liquid chromatography. Sphingolipids were quantified using mass spectrometry. Data were analyzed using a mixed model with repeated measures. Overweight cows had a higher BCS and BW at enrollment relative to lean cows, but BCS and BW were similar postpartum. Overweight cows lost more body condition (0.97 ± 0.36 vs. 0.55 ± 0.16 BCS units) and BW (291 ± 67.3 vs. 202 ± 54.5 kg) during transition relative to lean cows. Adipocyte volume and counts declined from prepartum to postpartum (50.4 and 13.7%, respectively), and adipocyte volume was greater (48.2%) in overweight cows prepartum relative to lean cows. Although DMI was comparable between BCS groups, milk yield tended to be greater in overweight cows. Plasma free fatty acid and ß-hydroxybutyrate and liver lipid levels were 40, 16, and 37% greater, respectively, in overweight cows compared with lean cows. Glucose infusion rate during the hyperinsulinemic-euglycemic clamp tended to be lower in overweight cows. Ceramide levels within triacylglycerol-rich lipoprotein fractions declined postpartum, whereas LDL ceramide increased postpartum. Overweight cows had lower triacylglycerol-rich lipoprotein C16:0-ceramide levels relative to lean cows. Prepartum LDL C24:0-ceramide levels were greater in overweight cows relative to lean cows. Independent of prepartum adiposity, we concluded that serum LDL ceramide levels are elevated in early-lactation cows experiencing adipose tissue free fatty acid mobilization and hepatic steatosis.

Methyl donor supplementation suppresses the progression of liver lipid accumulation while modifying the plasma triacylglycerol lipidome in periparturient Holstein dairy cows.

Co-supplementation of methyl donors may lower hepatic lipid content in transition cows. To define the ability of methyl donor supplementation (MDS) to reduce hepatic lipid content and modify the plasma lipidome, 30 multiparous Holstein cows (2.04 ± 0.69 lactations; 689 ± 58 kg of body weight; 3.48 ± 0.10 units of body condition score) were fed a ration with or without rumen-protected methyl donors (22 g/d of Met, 10 g/d of choline chloride, 3 g/d of betaine, 96 mg/d of riboflavin, and 1.4 mg/d of vitamin B12) from d -28 before expected calving through d 14 postpartum. Cows were randomly enrolled based on predefined selection criteria (body condition score and parity). Base diets without MDS were formulated for gestation (15.4% crude protein with a predicted Lys-to-Met ratio of 3.25; 1.44 Mcal of net energy for lactation/kg of dry matter) and lactation (16.6% crude protein with a predicted Lys-to-Met ratio of 3.36; 1.64 Mcal of net energy for lactation/kg of dry matter). Blood sampling occurred from d -28 relative to expected calving through d 14 postpartum. Liver tissue was biopsied at d -28 relative to expected calving and on d 5 and 14 postpartum. In addition to routine analyses, serum AA concentrations on d 10 and 12 were quantified using mass spectrometry. Plasma triacylglycerol (TAG) and cholesteryl esters (CE) were qualitatively measured using time-of-flight mass spectrometry. Data were analyzed using a mixed model with repeated measures. Dry matter intake and milk yield were not modified by MDS. The transition from d -28 relative to expected parturition to d 14 postpartum was characterized by increased plasma fatty acid (0.15 to 0.71 mmol/L) and ß-hydroxybutyrate (0.34 to 0.43 mmol/L) levels and liver lipid content (3.91 to 9.16%). Methyl donor supplementation increased the serum Met level by 26% and decreased the serum Lys-to-Met ratio by 21% on d 10 and 12, respectively. Moreover, the increase in hepatic lipid content from d 5 through 14 postpartum was suppressed with MDS relative to control (3.57 vs. -0.29%). Dietary MDS modified the TAG and CE lipidome. For example, MDS increased plasma TAG 46:3 (carbon number:double bond) by 116% relative to control cows on d 5 postpartum. Moreover, MDS tended to increase plasma CE 34:6. In contrast, MDS lowered plasma TAG 54:8 by 39% relative to control cows on d 5 postpartum. We concluded that in the absence of gains in dry matter intake and milk and milk protein yields, dietary MDS slows the progression of hepatic lipid accumulation and modifies the plasma TAG lipidome in transition cows.

Hot topic: Ceramide inhibits insulin sensitivity in primary bovine adipocytes.

In nonruminants, the sphingolipid ceramide inhibits insulin sensitivity by inactivating protein kinase B (AKT) within the insulin-signaling pathway. We have established that ceramide accrual develops with impaired systemic insulin action in ruminants during the transition from gestation to lactation, dietary palmitic acid supplementation, or controlled nutrient restriction. We hypothesized that ceramide promotes AKT inactivation and antagonizes insulin sensitivity in primary bovine adipocytes. Stromal-vascular cells were grown from bovine adipose tissue explants and cultured in differentiation media. To modify ceramide supply, we treated differentiated adipocytes with (1) myriocin, an inhibitor of de novo ceramide synthesis, or (2) cell-permeable C2:0-ceramide. Insulin-stimulated AKT activation (i.e., phosphorylation) and 2-deoxy-D-[3H]-glucose (2DOG) uptake were measured. Treatment of adipocytes with myriocin consistently decreased concentrations of ceramide, monohexosylceramide, and lactosylceramide. The insulin-stimulated ratio of phosphorylated AKT to total AKT was increased with myriocin but decreased with C2:0-ceramide. Moreover, adipocyte insulin-stimulated 2DOG uptake was decreased with C2:0-ceramide and increased with myriocin. We conclude that ceramide inhibits insulin-stimulated glucose uptake by downregulating AKT activation in primary bovine adipocytes.

Spin Resonance Clock Transition of the Endohedral Fullerene

The endohedral fullerene ^{15}N@C_{60} has narrow electron paramagnetic resonance lines which have been proposed as the basis for a condensed-matter portable atomic clock. We measure the low-frequency spectrum of this molecule, identifying and characterizing a clock transition at which the frequency becomes insensitive to magnetic field. We infer a linewidth at the clock field of 100 kHz. Using experimental data, we are able to place a bound on the clock's projected frequency stability. We discuss ways to improve the frequency stability to be competitive with existing miniature clocks.

Subtype associations with HIV-associated neurocognitive disorder in China.

Factors associated with HIV-associated neurocognitive disorders (HAND) include CD4(+) nadir and count, HIV RNA level, and HIV-1 subtype. Here, we investigated demographical and clinical markers with respect to HAND in a homogenous Chinese population. Individuals with HAND (global deficit score ≥0.5) had lower nadir (p < 0.01) and CD4(+) counts (p = 0.03). HAND was also associated with AIDS (p < 0.01), but subtype was not (p = 0.198). Furthermore, worse impairment correlated with higher viral diversity (r = 0.16, p < 0.01), lower nadir (r = -0.17, p < 0.01), and CD4(+) counts (r = -0.11, p = 0.01). These remained significant even when correcting for subtype. Our findings suggest that subtype does not have a major impact on HAND.

The placebo as performance: speaking across domains of healing.

In this article I explore the placebo and the placebo effect from a performance-studies perspective. I use this examination to argue for the placebo as a possible dialogic starting point between performance studies and qualitative health scholars. Using Lock and Scheper-Hughes' concept of three bodies (the individual body, the social body, and the body politic), I explain how the placebo as performance opens dialogue by speaking across these three bodies. I argue that the placebo as performance offers a yet-unexplored and heuristic way to bridge the unfortunate divide that often exists between qualitative and quantitative ways of understanding healing. By exploring these connections and offering a history of blind testing in the medical community, I explain how the placebo requires multiple lenses to be understood in the healing process, and by extension opens up traffic between different ways of knowing.

The local immune response in ulcerative lesions of Buruli disease.

Buruli disease (BU) is a progressive necrotic and ulcerative disease of the skin and subcutaneous tissue caused by Mycobacterium ulcerans. BU is considered the third most common mycobacterial disease after tuberculosis and leprosy. Three clinical stages of the cutaneous lesions have been described in BU: pre-ulcerative, ulcerative and healed lesions. In this study we used immunohistochemistry and automated morphometry to determine the percentage of macrophages and of CD4/CD8 lymphocytes and their expression of interferon (IFN)-gamma, interleukin (IL)-10, tumour necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta. Expression of these cytokines was correlated with the inflammatory response evaluated by histopathology. All the studied BU ulcerative cases showed extensive necrosis and chronic inflammation. The most important feature was the presence or absence of granulomas co-existing with a mixed pro-inflammatory/anti-inflammatory cytokine balance. When granulomas were present significantly higher expression of IFN-gamma was seen, whereas in ulcerative lesions without granulomas there was increased expression of IL-10 and significantly higher bacillary counts. These features correlated with the chronicity of the lesions; longer-lasting lesions showed granulomas. Thus, granulomas were absent from relatively early ulcerative lesions, which contained more bacilli and little IFN-gamma, suggesting that at this stage of the disease strong suppression of the protective cellular immune response facilitates proliferation of bacilli.

Technical note: a procedure for the preparation and quantitative analysis of samples for titanium dioxide.

A procedure was developed for the rapid analysis of titanium dioxide (TiO2) concentrations in feed and fecal samples. Samples were digested in concentrated H2SO4 for 2 h, followed by addition of 30% H2O2, and absorbance was measured at 410 nm. Standards were prepared by spiking blanks with increasing amounts of TiO2, resulting in a linear standard curve. Complete analysis using this procedure can typically be accomplished within 4.5 h. This procedure was compared to a previously published dry-ash procedure for the analysis of TiO2 in bovine fecal samples. Three sources of OM devoid of TiO2 (a forage sample, a bovine fecal sample without Cr2O3, and a bovine fecal sample containing Cr2O3) were spiked with graded amounts (0, 2, 4, 6, 8, or 10 mg) of TiO2. With our procedure, TiO2 recoveries averaged 96.7, 97.5, and 98.5%, for the three OM sources, respectively, vs. 74.3, 83.8, and 53.1% for the same samples analyzed using the dry-ash method. These results suggest that our procedure is a rapid and accurate alternative to dry-ash procedures for the determination of TiO2.

Detection of bacteria in suspension by using a superconducting quantum interference device.

We demonstrate a technique for detecting magnetically labeled Listeria monocytogenes and for measuring the binding rate between antibody-linked magnetic particles and bacteria. This sensitive assay quantifies specific bacteria in a sample without the need to immobilize them or wash away unbound magnetic particles. In the measurement, we add 50-nm-diameter superparamagnetic magnetite particles, coated with antibodies, to an aqueous sample containing L. monocytogenes. We apply a pulsed magnetic field to align the magnetic dipole moments and use a high-transition temperature superconducting quantum interference device, an extremely sensitive detector of magnetic flux, to measure the magnetic relaxation signal when the field is turned off. Unbound particles randomize direction by Brownian rotation too quickly to be detected. In contrast, particles bound to L. monocytogenes are effectively immobilized and relax in about 1 s by rotation of the internal dipole moment. This Néel relaxation process is detected by the superconducting quantum interference device. The measurements indicate a detection limit of (5.6 +/- 1.1) x 10(6) L. monocytogenes in our sample volume of 20 microl. If the sample volume were reduced to 1 nl, we estimate that the detection limit could be improved to 230 +/- 40 L. monocytogenes cells. Time-resolved measurements yield the binding rate between the particles and bacteria.

Improving performance at the local level: implementing a public health learning workforce intervention.

In an effort to continually improve performance of the essential public health services with community partners, the diverse public health workforce in a major metropolitan area engaged in an organizational learning process. Core public health organizational competencies, identified in a multi-year collaborative applied research initiative, provided the curricula content for the public health learning experience. All members (about 600) of the Columbus and Franklin County (Ohio) Health Departments participated in four one-half day small group, highly interactive modules conducted during a 2-year period. The purpose of this article is to describe the design and implementation of this workforce intervention, the lessons learned, and implications for developing organizational capacity and improved performance.

The in-vitro porcine adhesion model is not predictive of the esophageal transit of risedronate tablets in humans.

Mucosal damage due to esophageal adhesion of pharmaceuticals is a continued concern to both health care providers and drug manufacturers. As a result of this concern, dosage forms are now being designed to exhibit minimal esophageal adhesion. Previous researchers have used an in-vitro porcine esophageal model to determine the propensity for formulations to adhere to the esophagus as an alternative to human scintigraphy studies. This study used a porcine esophageal adhesion model similar to that used previously to determine the adhesiveness of placebo bisphosphonate formulations. Results are analogous to those obtained by previous researchers, with film-coated tablets showing greater adhesiveness than uncoated tablets. These same tablet formulations were also evaluated previously by a human scintigraphy study, and the results were exactly opposite of those obtained using the in-vitro porcine model. In the human scintigraphy study, the film-coated placebo risedronate tablet had a faster transit time than an uncoated round placebo tablet. In conclusion, the in-vitro porcine esophageal model is not predictive of esophageal transit in man and gamma scintigraphy is the preferred method to evaluate esophageal transit.

Autocastration as a presenting sign of incipient schizophrenia.

Previous case reports on autocastration have identified individuals who engaged in genital self-mutilation as being overtly psychotic, as having previously received a diagnosis of schizophrenia, or as having significant dysfunction of ego integrity, manifesting as guilt and low self-esteem. This paper describes a patient who had had no previous psychiatric symptoms or mental health treatment and for whom the act of autocastration was the first sign of incipient schizophrenia.

Detection of diagnostically critical, often hidden, anomalies in complex karyotypes of haematological disorders using multicolour fluorescence in situ hybridization.

Multicolour fluorescence in situ hybridization (M-FISH) simultaneously detects all 24 human chromosomes in unique fluorescent colours. The identification of diagnostically critical gene rearrangement(s) in complex karyotypes of haematological disorders continues to be a challenge. We present five cases in which t(9;11), complex t(8;22), t(12;21) and t(11;14) were detected primarily using M-FISH and were confirmed using locus-specific probes. We conclude that M-FISH can be effective in complete characterization of critical gene rearrangements in haematological disorders.