Feeding rumen-protected lysine prepartum alters placental metabolism at a transcriptional level.

Rumen-protected Lys (RPL) fed to Holstein cows prepartum resulted in a greater intake and improved health of their calves during the first 6 wk of life. However, whether increased supply of Lys in late gestation can influence placental tissue and, if so, which pathways are affected remain to be investigated. Therefore, we hypothesize that feeding RPL during late gestation could modulate placental metabolism, allowing for improved passage of nutrients to the fetus and thus influencing the offspring development. Therefore, we aimed to determine the effects of feeding RPL (AjiPro-L Generation 3, Ajinomoto Health and Nutrition North America) prepartum (0.54% DM of TMR) on mRNA gene expression profiles of placental samples of Holstein cows. Seventy multiparous Holstein cows were randomly assigned to 1 of 2 dietary treatments, consisting of TMR top-dressed with RPL (PRE-L) or without (control, CON), fed from 27 ± 5 d prepartum until calving. After natural delivery (6.87 ± 3.32 h), placentas were rinsed with physiological saline (0.9% sodium chloride solution) to clean any dirtiness from the environment and weighed. Then, 3 placentomes were collected, one from each placental region (cranial, central, and caudal), combined and flash-frozen in liquid nitrogen to evaluate the expression of transcripts and proteins related to protein metabolism and inflammation. Placental weights did not differ from cows in PRE-L (15.5 ± 4.03 kg) and cows in CON (14.5 ± 4.03 kg). Feeding RPL prepartum downregulated the expression of NOS3 (nitric oxide synthase 3), involved in vasodilation processes, and SOD1, which encodes the enzyme superoxide dismutase, involved in oxidative stress processes. Additionally, feeding RPL prepartum upregulated the expression of transcripts involved in energy metabolism (SLC2A3, glucose transporter 3; and PCK1, phosphoenolpyruvate carboxykinase 1), placental metabolism and cell proliferation (FGF2, fibroblast growth factor 2; FGF2R, fibroblast growth factor 2 receptor; and PGF, placental growth factor), Met metabolism (MAT2A, methionine adenosyltransferase 2-α), and tended to upregulate IGF2R (insulin-like growth factor 2 receptor). Placental FGF2 and LRP1 (low-density lipoprotein receptor-related protein 1) protein abundance were greater for cows that received RPL prepartum than cows in CON. In conclusion, feeding RPL to prepartum dairy cows altered uteroplacental expression of genes and proteins involved in cell proliferation, and in metabolism and transport of glucose. Such changes are illustrated by increased expression of SLC2A3 and PCK1 and increased protein abundance of FGF2 and LRP1 in uteroplacental tissue of cows consuming RPL.

Feeding rumen-protected lysine altered immune and metabolic biomarkers in dairy cows during the transition period.

This experiment was conducted to determine the effects of feeding rumen-protected lysine (RPL; AjiPro-L Generation 3, Ajinomoto Health and Nutrition North America Inc.) from -26 ± 4.6 d prepartum (0.54% RPL of dietary dry matter intake) to 28 d postpartum (0.39% RPL of dietary dry matter intake) on immunometabolic status and liver composition in dairy cows. Seventy-five multiparous Holstein cows, blocked by parity, previous 305-d mature-equivalent milk production, expected calving date, and body condition score during the far-off dry period were assigned to 1 of 4 dietary treatments in a randomized, complete block design with a 2 × 2 factorial arrangement of treatments. Treatments prepartum consisted of total mixed ration top dressed with RPL (PRE-L) or without RPL (PRE-C), and postpartum treatments consisted of total mixed ration top dressed PRE-L prepartum and postpartum, PRE-L prepartum and PRE-C postpartum, PRE-C prepartum and PRE-L postpartum, and PRE-C prepartum and postpartum in 300 g of molasses. Blood samples were taken on -7 ± 0.5, 0 ± 0.5, 7 ± 0.9, 14 ± 0.9, and 28 ± 0.5 d relative to calving. Whole blood samples were taken on -14 ± 0.5, -7 ± 0.5, 7 ± 0.9, and 14 ± 0.9 d relative to calving for oxidative burst and phagocytic capacity of monocytes and neutrophils. Liver samples were collected via a biopsy on -12 ± 4.95 and 13 ± 2.62 d relative to calving and analyzed for liver composition (triacylglyceride and carnitine concentrations), mRNA expression of hepatic genes, and protein abundance. Protein abundance was calculated by normalizing intensity bands for a specific protein with glyceraldehyde-3-phosphate dehydrogenase. Concentrations of haptoglobin and glutathione peroxidase activity in plasma were lower at d 0 for cows in PRE-L (102 µg/mL and 339 nmol/min per mL, respectively) compared with cows in PRE-C (165 µg/mL and 405 nmol/min per mL, respectively). Oxidative burst capacity in monocytes tended to be greater on d 7 postpartum for cows in PRE-L (65.6%) than cows in PRE-C (57.5%). Additionally, feeding RPL altered the mRNA expression in liver tissue prepartum [decreased INSR (insulin receptor), CPT1A (carnitine palmitoyltransferase 1A), and IL1B (interleukin 1 ß)] and postpartum [increased IL8 (interleukin 8), EHMT2 (euchromatic histone lysine methyltransferase 2), TSPO (translocator protein), and SLC3A2 (solute carrier family 3 member 2); and decreased SLC7A1 (solute carrier family 7 member 1), SOD1 (superoxide dismutase 1), and SAA3 (serum amyloid A 3)] compared with cows not consuming RPL]. Additionally, cows in the PRE-C prepartum and PRE-L postpartum treatment tended to have greater protein abundance of mTOR postpartum compared with the PRE-C prepartum and postpartum treatment. Protein abundance of SLC7A7 (solute carrier family 7 member 7) pre- and postpartum tended to be greater and BBOX1 (gamma-butyrobetaine dioxygenase 1) tended to be less when RPL was consumed prepartum. In conclusion, cows that consumed RPL during the transition period had molecular changes related to liver composition, enhanced liver function indicated by greater total protein and albumin concentrations in plasma, and improved immune status indicated by decreased haptoglobin, glutathione peroxidase activity, and immune related mRNA expression.

Effect of feeding rumen-protected lysine through the transition period on postpartum uterine health of dairy cows.

Feeding rumen-protected methionine as an indispensable amino acid source has been shown to improve reproductive performance in dairy cows, but the effect of feeding rumen-protected lysine (RPL) during the peripartum period on reproductive performance is not well explored. Therefore, we aimed to determine the effects of feeding RPL (AjiPro-L Generation 3, Ajinomoto Heartland Inc.) prepartum, postpartum, or both on follicular dynamics, uterine health, and mRNA gene expression of the endometrium. Seventy-five multiparous Holstein cows were assigned to 1 of 2 dietary treatments with or without RPL in a randomized, complete block design. A 2 × 2 factorial arrangement of treatments was used. Prepartum (-28 d to calving), animals were fed a diet (68% of dietary DM from forage) with RPL [PRE-L; 0.54% RPL of dietary dry matter intake] or without RPL (PRE-C). After calving, half of the cows from each prepartum treatment group were assigned to a diet (56% forage) with RPL (PRE-L POST-L; PRE-C POST-L; 0.40% RPL of dietary dry matter intake) or without RPL (PRE-C POST-C; PRE-L POST-C) until 28 d in milk (DIM). Vaginal discharge was detected with a Metricheck device (Simcro) to detect metritis, and at 28 DIM polymorphonuclear leukocytes were evaluated as a percentage of the epithelial cells using a cytology brush (Andwin Scientific) and an endometrial tissue biopsy was collected for mRNA expression and histology. The first postpartum follicular growth cycle was monitored at 7, 10, 11, 13, 15, 17, 19, 21, 23, 25, 27, and 28 DIM via transrectal ultrasonography. Time to first ovulation did not differ between treatments and averaged 18 ± 1.6 DIM. Follicular diameter at first ovulation was not affected by the treatments, but the growth rate of dominant follicle before first ovulation tended to be lower for cows in POST-L in comparison with cows in POST-C. Prevalence of fetid vaginal discharge and metritis did not differ between treatments. Cows in PRE-L POST-L had lower polymorphonuclear leukocytes percentage at 15 and 28 DIM than cows in PRE-L POST-C, PRE-C POST-L, and PRE-C POST-C. Feeding RPL prepartum downregulates the expression of TLR4, SLC7A6, EHMT2, and tends to downregulate the expression of PTGES3 in uterine tissues at 28 DIM. Additionally, it upregulates the expression of APOL3 and NFKB1, and tends to upregulate the expression of AHCY and MAT2A. In conclusion, feeding RPL pre- and postpartum improved indicators of uterine immune status, but did not change days to first ovulation postpartum.

Feeding rumen-protected lysine to dairy cows prepartum improves performance and health of their calves.

Providing adequate concentrations of AA in the prepartum diet is pivotal for the cow's health and performance. However, less is known about the potential in utero effects of particular AA on early-life performance of calves. This experiment was conducted to determine the effects on dairy calves when their dams were fed rumen-protected lysine (RPL; AjiPro-L Generation 3, Ajinomoto Heartland Inc.; 0.54% dry matter of total mixed ration as top dress) from 26 ± 4.6 d (mean ± standard deviation) before calving until calving. Seventy-eight male (M) and female (F) Holstein calves were assigned to 2 treatments based on their dams' prepartum treatment, RPL supplementation (PRE-L) or without RPL (CON). At the time of birth (0.5-2 h after calving), before colostrum was fed, blood samples were collected. An initial body weight was obtained at 1 to 3 h after birth. Calves were fed 470 g of colostrum replacer (Land O'Lakes Bovine IgG Colostrum Replacer, Land O'Lakes, Inc.) diluted in 3.8 L of water. Calves were provided water ad libitum and fed milk replacer (Advance Excelerate, Milk Specialties Global Animal Nutrition; 28.5% crude protein, 15% fat) at 0600 h and 1700 h until 42 d of age. Calves were measured weekly, at weaning (d 42), and at the end of the experimental period (d 56). Plasma concentrations of AA were measured on d 0, 7, and 14 d using ultra-performance liquid chromatography-mass spectrometry (Waters) with a derivatization method (AccQ-Tag Derivatization). Final body weight was greater for M (87 ± 11 kg) than F (79 ± 7 kg). Calves in PRE-L tended to have greater dry matter (814 ± 3 g/d) and crude protein (234 ± 6 g/d) intakes than those in CON (793 ± 9 g/d and 228 ± 11 g/d, respectively). Calves in PRE-L had greater average daily gain (0.96 ± 0.04 kg/d) than calves in CON (0.85 ± 0.03 kg/d) during wk 6 to 8. Calves in PRE-L tended to be medicated fewer days than CON (4.7 ± 1.2 d vs. 6.2 ± 3.4 d, respectively). Calves in PRE-L-M and CON-F (2,916 ± 112 µM and 2,848 ± 112 µM, respectively) had greater total AA concentration in plasma than calves in PRE-L-F and CON-M (2,684 ± 112 µM and 2,582 ± 112 µM, respectively). Calves in PRE-L-F and CON-M (4.09 ± 0.11% and 4.16 ± 0.11%, respectively) had greater concentration of Lys as a percentage of total AA compared with calves in CON-F and PRE-L-M (3.91 ± 0.11% and 3.90 ± 0.11%, respectively). Calves in PRE-L tended to have greater percentage of phagocytic neutrophils (39.6 ± 1.59%) than calves in CON (35.9 ± 1.59%). In conclusion, increasing the metabolizable lysine provided to prepartum dairy cows had modest effect over offspring performance, with the major result being a greater average daily gain for calves in PRE-L during the preweaning phase (wk 6-8).

Measurement of axial phase difference of density fluctuations owing to spontaneously excited waves by using microwave reflectometer on GAMMA 10/PDX.

In the GAMMA 10/PDX tandem mirror, plasma with strong ion-temperature anisotropy is produced by using the ion cyclotron range of frequency waves. This anisotropy of ion temperature causes several Alfvén-Ion-Cyclotron (AIC) waves to spontaneously excite in the frequency range just below the ion cyclotron frequency. In addition, difference-frequency (DF) waves are excited in the radial inner region of the plasma by wave-wave coupling among the AIC waves. The radial density profiles were measured at multi-axial positions using a frequency-modulation reflectometer with an axial array of microwave antennas, and an axial variation of the density was found to be significant. In addition, a relative phase difference of the DF wave between axially separated two points was first obtained by finely choosing the probing frequency of the reflectometers with a maximum coherence used as a measure, indicating that the DF wave is a propagating wave, while the pump AIC waves are standing waves in the axial region of measurement.

Feeding rumen-protected lysine prepartum increases energy-corrected milk and milk component yields in Holstein cows during early lactation.

Feeding rumen-protected Lys (RPL) may be used to increase lactation performance in dairy cows; however, the effect of feeding RPL during the prepartum period and subsequent effect on postpartum performance is not well explored. Therefore, this experiment was conducted to determine the effects of feeding RPL (AjiPro-L Generation 3, Ajinomoto Heartland Inc., Chicago, IL) prepartum, postpartum, or both on performance, health, and blood metabolites. Seventy-five multiparous Holstein cows, blocked by parity, previous 305-d mature-equivalent milk production, expected calving date, and body condition score during the far-off dry period were assigned to 1 of 2 dietary treatments: total mixed ration with or without RPL in a randomized, complete block design. A 2 × 2 factorial arrangement of treatments was used. Prepartum (-28 d to calving), animals were fed a diet (forage, 68% of dietary DM) with RPL [PRE-L; 0.54% RPL of dietary dry matter intake (DMI)] or without RPL (control; PRE-C). After calving, half of the cows from each prepartum treatment group were assigned to a diet (forage, 55.5% of dietary DM) with RPL (PRE-L POST-L; PRE-C POST-L; 0.40% RPL of dietary DMI) or without RPL (PRE-C POST-C; PRE-L POST-C) until d 28 postpartum. Cows were milked twice a day and milk samples were taken on 7 ± 1.3, 14 ± 1.4, and 28 ± 1.1 d relative to calving (DRC). Milk yield and DMI were recorded daily. Blood samples were taken for plasma AA analysis on -7 ± 0.5, 0 ± 0.5, 7 ± 0.9, and 14 ± 0.9 DRC. Cows in PRE-L had greater body weight at -2 and -1 wk before calving compared with those in PRE-C, though body weight change from wk -4 to -1 was not different. Body weight (717 ± 6 kg) was greater and DMI (18.1 ± 0.7 kg) tended to be greater for cows in PRE-L POST-L and PRE-L POST-C compared with those that were in PRE-C POST-L and PRE-C POST-C (707 ± 6 and 16.8 ± 0.7 kg, respectively). Energy-corrected milk (48.8 ± 1.9 kg/d), milk fat (1.9 ± 0.1 kg/d), milk true protein (1.4 ± 0.1 kg/d), milk casein (0.6 ± 0.04 kg/d), and milk lactose yields (2.1 ± 0.1 kg/d) were greater for cows in PRE-L POST-L and PRE-L POST-C compared with those that were in PRE-C POST-L and PRE-C POST-C (44.2 ± 1.9, 1.7 ± 0.1, 1.3 ± 0.1, 0.5 ± 0.04, 1.9 ± 0.1 kg/d, respectively). Plasma concentrations of Lys prepartum (69.8 ± 1.8 µM) increased for cows in PRE-L compared with those in PRE-C (62.5 ± 1.3 µM). In conclusion, RPL consumed prepartum tended to increase postpartum DMI and increased energy-corrected milk and milk component yields. This indicates that prepartum supply of intestinally available Lys is pertinent to postpartum performance. However, postpartum supply of intestinally available Lys had no effect on cows' performance.

Intrinsic reconstruction of ice-I surfaces.

Understanding the precise atomic structure of ice surfaces is critical for revealing the mechanisms of physical and chemical phenomena at the surfaces, such as ice growth, melting, and chemical reactions. Nevertheless, no conclusive structure has been established. In this study, noncontact atomic force microscopy was used to address the characterization of the atomic structures of ice Ih(0001) and Ic(111) surfaces. The topmost hydrogen atoms are arranged with a short-range (2 × 2) order, independent of the ice thickness and growth substrates used. The electrostatic repulsion between non-hydrogen-bonded water molecules at the surface causes a reduction in the number of the topmost hydrogen atoms together with a distortion of the ideal honeycomb arrangement of water molecules, leading to a short-range-ordered surface reconstruction.

Realization of Spin-dependent Functionality by Covering a Metal Surface with a Single Layer of Molecules.

An interface of molecule and metal has attracted much attention in the research field of nanoelectronics because of their high degree of design freedom. Here, we demonstrate an efficient spin-to-charge current conversion at the metal surface covered by a single layer of molecules. Spin currents are injected into an interface between metal (Cu) and lead(II) phthalocyanine by means of the spin pumping method. An observed voltage signal is caused by the inverse Edelstein effect, i.e., spin-to-charge current conversion at the interface. The conversion coefficient, inverse Edelstein length, is estimated to be 0.40 ± 0.06 nm, comparable with the largest Rashba spin splitting of interfaces with heavy metals. Interestingly, the Edelstein length strongly depends on the thickness of the molecule and takes a maximum value when a single layer of molecules is formed on the Cu surface. Comparative analysis between scanning probe microscopy and first-principles calculations reveal that the formation of interface state with Rashba spin splitting causes the inverse Edelstein effect, whose magnitude is sensitive to the adsorption configuration of the molecules.

Direct penetration of spin-triplet superconductivity into a ferromagnet in Au/SrRuO3/Sr2RuO4 junctions.

Efforts have been ongoing to establish superconducting spintronics utilizing ferromagnet/superconductor heterostructures. Previously reported devices are based on spin-singlet superconductors (SSCs), where the spin degree of freedom is lost. Spin-polarized supercurrent induction in ferromagnetic metals (FMs) is achieved even with SSCs, but only with the aid of interfacial complex magnetic structures, which severely affect information imprinted to the electron spin. Use of spin-triplet superconductors (TSCs) with spin-polarizable Cooper pairs potentially overcomes this difficulty and further leads to novel functionalities. Here, we report spin-triplet superconductivity induction into a FM SrRuO3 from a leading TSC candidate Sr2RuO4, by fabricating microscopic devices using an epitaxial SrRuO3/Sr2RuO4 hybrid. The differential conductance, exhibiting Andreev-reflection features with multiple energy scales up to around half tesla, indicates the penetration of superconductivity over a considerable distance of 15 nm across the SrRuO3 layer without help of interfacial complex magnetism. This demonstrates potential utility of FM/TSC devices for superspintronics.

Molecular cloning, polymorphism, and functional activity of the bovine and water buffalo Mx2 gene promoter region.

BACKGROUND: Bovine Mx2 gene sequences were already reported, but further information about the gene properties is not yet available. The objective of the current study was to elucidate the structural properties of the bovine Mx2 gene mainly the promoter region and its possible functional role. If available, such information would help in assessing the functional properties of the gene, which was reported to confer antiviral action against recombinant VSV. RESULTS: Examinations on the bovine genomic BAC clone-confirmed to contain the Mx2 gene-revealed 883-bp sequences. A computer scan unequivocally identified a 788-bp promoter region containing a typical TATA box, three ISREs and other promoter-specific motifs. Comparative analysis of nine bovine genomic DNA samples showed 19 nucleotide substitutions suggesting the existence of five different genotypes in the promoter region. The water buffalo Mx2 promoter region was determined by using primers based on the bovine Mx2 promoter region disclosing 893-bp, with 56 substitutions, two insertions, 9 and 1 nt at two different sites. A functional analysis of the putative ISRE indicated that ISRE played a synergetic role in the activation of bovine Mx2 gene transcription. CONCLUSION: Bovine and water buffalo Mx2 promoter region was identified disclosing, the conserved ISRE, located in the proximal end of the promoter region like other members of the antiviral family, suggesting functional activity under interferon stimulation.

Accuracy of high-density genotype imputation in Japanese Black cattle.

Genotype imputation facilitates the identification of missing genotypes on a high-density array using low-density arrays and has great potential for reducing genotyping costs for cattle populations. However, the imputation quality varies across breeds, which have different effective population sizes. Therefore, the accuracy of genotype imputation must be evaluated in each breed. The Japanese Black cattle population has a unique genetic background, and this study aimed to investigate different factors affecting imputation quality in this population. A total of 1368 animals were genotyped using the Illumina BovineHD BeadChip, and the accuracy of imputation was evaluated using information from four lower density arrays. The extent of linkage disequilibrium for this population was relatively higher than that in other beef breeds but lower than that in dairy breeds. The accuracy of arrays with more than 20 000 single nucleotide polymorphisms (SNPs) was similar to or higher than that of lower density arrays. In addition, the minor allele frequency of SNPs in the reference population affected the accuracy. The accuracy increased as the size of the reference population increased, up to 400 animals, beyond which there was little increase. A higher genetic relationship between the reference and test populations increased imputation accuracy. These results indicate that high imputation accuracy can be achieved using high-density arrays, having enough reference animals and including relatives in the reference population.

Impact of molecular mutations on treatment response to DNMT inhibitors in myelodysplasia and related neoplasms.

We hypothesized that specific molecular mutations are important biomarkers for response to DNA methyltransferase inhibitors (DNMT inhibitors) and may have prognostic value in patients with myelodysplastic syndromes (MDS). Mutational analysis was performed in 92 patients with MDS and related disorders who received 5-azacytidine (n=55), decitabine (n=26) or both (n=11). Mutational status was correlated with overall response rate (ORR), progression-free survival (PFS) and overall survival (OS) by univariate and multivariate analysis. Risk stratification models were created. TET2, DNMT3A, IDH1/IDH2, ASXL1, CBL, RAS and SF3B1 mutations were found in 18, 9, 8, 26, 3, 2 and 13% of patients, respectively. In multivariate analysis, TET2(MUT) and/or DNMT3A(MUT) (P=0.03), platelets > or = 100 × 10(9)/l (P=0.007) and WBC<3.0 × 10(9)/l (P=0.03) were independent predictors of better response. TET2(MUT) and/or DNMT3A(MUT) (P=0.04) status was also independently prognostic for improved PFS, as were good or intermediate cytogenetic risk (P<0.0001), age<60 (P=0.0001), treatment with both 5-azacytidine and decitabine (P=0.02) and hemoglobin > or = 10 g/dl (P=0.01). Better OS was associated with ASXL1(WT) (P=0.008) and SF3B1(MUT) (P=0.01), and, similar to PFS, cytogenetic risk (P=0.0002), age (P=0.02) and hemoglobin (P=0.04). These data support the role of molecular mutations as predictive biomarkers for response and survival in MDS patients treated with DNMT inhibitors.

Multiple mechanisms deregulate EZH2 and histone H3 lysine 27 epigenetic changes in myeloid malignancies.

Polycomb repressive complex 2 (PRC2) is involved in trimethylation of histone H3 lysine 27 (H3K27), chromatin condensation and transcriptional repression. The silencing function of PRC2 complex is mostly attributed to its intrinsic activity for methylating H3K27. Unlike in B-cell lymphomas, enhancer of zeste homolog 2 (EZH2) mutations in myeloid malignancies are inactivating/hypomorphic. When we assessed the mutational status in myeloid malignancies (N=469 cases examined), we found EZH2 and EED/SUZ12 mutations in 8% and 3.3% of cases, respectively. In addition to mutant cases, reduced EZH2 expression was also found in 78% cases with hemizygous deletion (-7/del7q cases involving EZH2 locus) and 41% of cases with diploid chromosome 7, most interestingly cases with spliceosomal mutations (U2AF1/SRSF2 mutations; 63% of cases). EZH2 mutations were characterized by decreased H3K27 trimethylation and increased chromatin relaxation at specific gene loci accompanied by higher transcriptional activity. One of the major downstream target is HOX gene family, involved in the regulation of stem cell self-renewal. HOXA9 was found to be overexpressed in cases with decreased EZH2 expression either by EZH2/spliceosomal mutations or because of -7/del7q. In summary, our results suggest that loss of gene repression through a variety of mutations resulting in reduced H3K27 trimethylation may contribute to leukemogenesis.

Intracerebral microinjection of interleukin-4/interleukin-13 reduces ß-amyloid accumulation in the ipsilateral side and improves cognitive deficits in young amyloid precursor protein 23 mice.

We previously reported that the anti-inflammatory cytokine interleukin (IL)-4 induced selective clearance of oligomeric ß-amyloid (Aß(1-42)) in rat primary type 2 microglial cells. For the present study, we investigated whether IL-4 and IL-13 could activate microglial cells to induce Aß clearance in vivo and improve cognitive deficits in APP23 mice, which are amyloid precursor protein transgenic mice. We administered an intracerebral microinjection of a mixture of IL-4 and IL-13 or of saline vehicle into one hemisphere of APP23 mice and their wild-type littermates, 4.5 and 9 months old, after which we evaluated the effects of these treatments on spatial learning and memory by Morris Water Maze test and on accumulated amounts of Aß. The cytokine injection significantly improved memory deficits of 4.5-month-old APP23 mice, but did not do so in 9-month-old APP23 mice, even though similar Aß reductions were observed in both age groups of APP23 mice in the ipsilateral neocortex. The cytokine injection improved memory impairment of 9-month-old wild-type (WT) mice in the probe trial. Immunohistochemical analysis of the 4.5-month-old APP23 mice revealed the presence of increased numbers of microglial cells at 2 days after the cytokine injection. In addition to induced CD36 expression in the activated microglia, increased expression of neprilysin, mainly in neurons, suggested that the cytokines improved the cognitive deficits via degradation and clearance of intra- and extraneuronal Aß peptides, of buffer-extractable nonplaque form. Double immunostaining also revealed that most of the activated microglia had the M2-like phenotype. This unique mechanism of IL-4/IL-13-induced clearance of Aß may provide an additional strategy to prevent and/or cure Alzheimer's disease at early stage.

Identification of an FBN1 mutation in bovine Marfan syndrome-like disease.

Mutations in the gene encoding fibrillin-1 (FBN1), a component of the extracellular microfibril, cause Marfan syndrome (MFS). Frequent observation of cattle with a normal withers height, but lower body weight than age-matched normal cattle, was recently reported among cattle sired by phenotypically normal Bull A, in Japanese Black cattle. These cattle also showed other characteristic features similar to the clinical phenotype of human MFS, such as a long phalanx proximalis, oval face and crystalline lens cloudiness. We first screened a paternal half-sib family comprising 36 affected and 10 normal offspring of Bull A using the BovineSNP50 BeadChip (illumina). Twenty-two microsatellite markers mapped to a significant region on BTA10 were subsequently genotyped on the family. The bovine Marfan syndrome-like disease (MFSL) was mapped onto BTA10. As FBN1 is located in the significant region, FBN1 was sequenced in Bull A, and three affected and one normal cattle. A G>A mutation at the intron64 splicing accepter site (c.8227-1G>A) was detected in 31 of 36 affected animals (84.7%). The c.8227-1G>A polymorphism was not found in 20 normal offspring of Bull A or in 93 normal cattle unrelated to Bull A. The mutation caused a 1-base shift of the intron64 splicing accepter site to the 3' direction, and a 1-base deletion in processed mRNA. This 1-base deletion creates a premature termination codon, and a 125-amino acid shorter Fibrillin-1 protein is produced from the mutant mRNA. We therefore conclude that the c.8227-1G>A mutation is causative for MFSL. Furthermore, it was suggested that Bull A exhibited germline mosaicism for the mutation, and that the frequency of the mutant sperm was 14.9%.

CBL mutation-related patterns of phosphorylation and sensitivity to tyrosine kinase inhibitors.

Recurrent homozygous CBL-inactivating mutations in myeloid malignancies decrease ubiquitin ligase activity that inactivates SRC family kinases (SFK) and receptor tyrosine kinases (RTK). However, the most important SFK and RTK affected by these mutations, and hence, the most important therapeutic targets, have not been clearly characterized. We compared SFK and RTK pathway activity and inhibitors in acute myeloid leukemia cell lines containing homozygous R420Q mutation (GDM-1), heterozygous deletion (MOLM13) and wild-type (WT) CBL (THP1, U937). As expected with CBL loss, GDM-1 displayed high KIT expression and granulocyte-macrophage colony-stimulating factor (GM-CSF) hypersensitivity. Ectopic expression of WT CBL decreased GDM-1 proliferation but not cell lines with WT CBL. GDM-1, but not the other cell lines, was highly sensitive to growth inhibition by dasatinib (dual SFK and RTK inhibitor, LD50 50 nM); there was less or no selective inhibition of GDM-1 growth by sunitinib (RTK inhibitor), imatinib (ABL, KIT inhibitor), or PP2 (SFK inhibitor). Phosphoprotein analysis identified phosphorylation targets uniquely inhibited by dasatinib treatment of GDM-1, including a number of proteins in the KIT and GM-CSF receptor pathways (for example, KIT Tyr721, STAT3 Tyr705). In conclusion, the promiscuous effects of CBL loss on SFK and RTK signaling appear to be best targeted by dual SFK and RTK inhibition.

The SNP c.1326T>G in the non-SMC condensin I complex, subunit G (NCAPG) gene encoding a p.Ile442Met variant is associated with an increase in body frame size at puberty in cattle.

Recently, we had located a bovine carcass weight QTL, CW-2, to a 591-kb interval on BTA6 and have identified the SNP c.1326T>G in the NCAPG (non-SMC condensin I complex, subunit G) gene that leads to the amino acid change p.Ile442Met in the NCAPG protein, which is a candidate causative variation. Here, we examined the association of the NCAPG:c.1326T>G locus with linear skeletal measurements of growth-associated traits during adolescence, which is a period of intensive growth, using two historically and geographically distant cattle populations: 792 Japanese Black steers and 161 F(2) bulls of an experimental cross from Charolais and German Holstein. In both populations, the SNP NCAPG:c.1326T>G was associated with each component of body frame size: height, length and width at puberty. The associations of CW-2 with height- and length-associated traits were observed at an earlier growth period compared to the associations with thickness- and width-associated traits, indicating that the primary effect of the CW-2 QTL may possibly be exerted on skeletal growth. The significant associations of the NCAPG:c.1326T>G locus with growth-associated skeletal measurements are similar to the effects of the syntenic region on human chromosome 4 that are associated with adult height in humans, supporting the hypothesis that CW-2 is analogous to the human locus and pointing to a conserved growth-associated locus or chromosomal region present in both species.

Cavity formation on an optical nanofiber using focused ion beam milling technique.

We present the experimental realization of nanofiber Bragg grating (NFBG) by drilling periodic nano-grooves on a subwavelength-diameter silica fiber using focused ion beam milling technique. Using such NFBG structures we have realized nanofiber cavity systems. The typical finesse of such nanofiber cavity is F ∼ 20 - 120 and the on-resonance transmission is ∼ 30 - 80%. Moreover the structural symmetry of such NFBGs results in polarization-selective modes in the nanofiber cavity. Due to the strong confinement of the field in the guided mode, such a nanofiber cavity can become a promising workbench for cavity QED.

Enhanced clinical mastitis resistance in Holsteins with a FEZL p.Gly105(12_13) polymorphism.

Mastitis is a common infectious disease of the mammary gland and a major problem in the dairy industry. We previously reported that forebrain embryonic zinc finger-like (FEZL) encoding a stretch of 12 glycines (p.Gly105[12]) instead of 13 glycines (p.Gly105[13]) is associated with a lower somatic cell score (SCS) in a family derived from Walkway Chief Mark. Here we report that the p.Gly105[12] allele is associated with a significantly decreased incidence of clinical mastitis in a large Holstein population. We genotyped the FEZL polymorphism in 918 randomly collected Holstein sires, and investigated the effect of the polymorphism on the estimated breeding value (EBV) for SCS and milk, fat, solids-not-fat, and protein yield, and on the number of cattle with clinical mastitis among daughters derived from these sires. The average EBV for SCS among sires carrying the heterozygous p.Gly105[12] was significantly lower than that among sires carrying the homozygous p.Gly105[13], whereas we found no unfavorable effects of this polymorphism on EBV for milk, fat, solids-not-fat, and protein yield. The proportion of cows with clinical mastitis derived from sires carrying heterozygous p.Gly105[12] was significantly lower than that of daughters derived from sires carrying the homozygous p.Gly105[13]. Thus, selection of sires carrying p.Gly105[12] could be beneficial in the dairy industry by reducing the incidence of mastitis.