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).

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.

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.

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.

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%.

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.

Acceleration of intestinal polyposis through prostaglandin receptor EP2 in Apc(Delta 716) knockout mice.

Arachidonic acid is metabolized to prostaglandin H(2) (PGH(2)) by cyclooxygenase (COX). COX-2, the inducible COX isozyme, has a key role in intestinal polyposis. Among the metabolites of PGH(2), PGE(2) is implicated in tumorigenesis because its level is markedly elevated in tissues of intestinal adenoma and colon cancer. Here we show that homozygous deletion of the gene encoding a cell-surface receptor of PGE(2), EP2, causes decreases in number and size of intestinal polyps in Apc(Delta 716) mice (a mouse model for human familial adenomatous polyposis). This effect is similar to that of COX-2 gene disruption. We also show that COX-2 expression is boosted by PGE(2) through the EP2 receptor via a positive feedback loop. Homozygous gene knockout for other PGE(2) receptors, EP1 or EP3, did not affect intestinal polyp formation in Apc(Delta 716) mice. We conclude that EP2 is the major receptor mediating the PGE2 signal generated by COX-2 upregulation in intestinal polyposis, and that increased cellular cAMP stimulates expression of more COX-2 and vascular endothelial growth factor in the polyp stroma.

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.

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.

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.

NC-AFM imaging of the TiO(2)(110)-(1 x 1) surface at low temperature.

The TiO(2)(110)-(1 x 1) surface is investigated using non-contact atomic force microscopy (nc-AFM) at 80 K. We successfully obtained a distinct type of image contrast mode which does not exhibit hydroxyl (OH) impurity defects that mostly appear in common nc-AFM images. We named the obtained distinct type of image contrast as the 'hidden mode'. The assignments of surface atomic rows in this contrast mode are not easy in the absence of defects. By recording different contrast modes in the same region of the surface, we identified the atomic rows obtained in the 'hidden mode' image contrast as bridging oxygen atoms (O(b)). The mechanism of contrast formation was attributed to tip-induced displacement of H atoms over oxygen atoms in the OH groups on the O(b) rows. This interpretation was supported by dissipation measurements. A possible candidate for the tip-generating hidden-mode image contrast was interpreted to be a positively terminated tip apex with a dimer-like structure, revealing an attractive interaction with oxygen and a repulsive force on H atom sites. In addition, with a different tip state at close tip-sample distances, we were able to successfully resolve a high resolution image of the in-plane oxygen atoms.

Small-amplitude dynamic force microscopy using a quartz cantilever with an optical interferometer.

We propose dynamic force microscopy (DFM) that employs a quartz cantilever in combination with an interferometric deflection sensor. The high stiffness of the quartz cantilever allows DFM to be performed at oscillation amplitudes as small as 1 A. DFM topographic images can be obtained for a Si(111)-(7 x 7) surface even when a blunt tip is used at room temperature. The low-noise interferometer with a deflection noise floor of n = 15 fm Hz(-1/2) exhibits a high force sensitivity in force spectroscopy. By subtracting the huge van der Waals force contribution, we obtain a short-range force curve that is in good agreement with the theoretical curve of the covalent bonding force. Simultaneous measurement of the tunneling current during DFM imaging is also demonstrated.

Differentially expressed genes during bovine intramuscular adipocyte differentiation profiled by serial analysis of gene expression.

Beef marbling or intramuscular fat deposition is an economically important carcass trait in Japanese Black cattle. To investigate genes involved in intramuscular adipogenesis, differential gene expression during adipogenesis in a clonal bovine intramuscular preadipocyte (BIP) cell line was profiled with serial analysis of gene expression (SAGE). We sequenced 75 283 tags for the proliferation phase (day 0) and 81 878 tags for the differentiation phase (4 days after adipogenic stimulation: day 4). A comparison of the unique SAGE tag frequencies between the day 0- and day 4-libraries revealed that 878 (2.8%) of the 30 989 unique putative transcripts were expressed at significantly different levels (P < 0.05); 401 tags (1.4%) were up-regulated and 477 tags (1.2%) were down-regulated in the day 4-library relative to the day 0-library. We confirmed up-regulation of 10 tags of the genes that were up-regulated in the previous subtraction cloning studies in BIP cells [Animal Science Journal, 76 (2005) 479]. Of the 878 differentially expressed tags, 377 were identified in the bovine RefSeq library and 356 were assigned a bovine draft genomic sequence. Fifteen tags were mapped in previously detected beef marbling quantitative trait loci (QTL) regions [Mammalian Genome, 18 (2007) 125]. These genes may be involved in the adipogenic processes of beef marbling.

Percutaneous screw fixation for traumatic spondylolisthesis of the axis using iso-C3D fluoroscopy-assisted navigation (case report).

INTRODUCTION: Stable hangman's fractures are usually treated with a Halo vest fixation; however, this is not always effective in patients with polytrauma. These patients benefit from minimally invasive surgery because it allows for early rehabilitation and reduced nursing care. This is the case report on percutaneous screw fixation using three dimensional fluoroscopy-assisted navigation in a patient with polytrauma and a hangman's fracture. CASE REPORT: The patient was a 69-year-old woman involved in a traffic accident. Radiographs and CT showed bilateral fractures through the neural arch at the base of the C2 pedicles. External immobilization was difficult due to her polytrauma. INTERVENTION: A dynamic reference arc was attached to the spinous process of the axis through a small incision. After image acquisition, the fluoroscope workstation generated 3-dimensional reconstructions of the imaged anatomy. We made two small, lateral incisions for percutaneous screw insertion, and used an image-guided awl to create screw holes. A guide-wire was inserted through this screw pilot hole, and a cancellous lag screw was inserted over the guide-wire. At her final follow-up, the patient had no neurological deficits and bony union was achieved. CONCLUSION: Percutaneous screws fixation using three-dimensional fluoroscopy proved to be a useful technique for the treatment of hangman's fracture.

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.

Colocalization of prostaglandin F(2alpha) receptor FP and prostaglandin F synthase-I in the spinal cord.

Prostaglandin F(2alpha) is synthesized by prostaglandin F synthase, which exists in two types, prostaglandin F synthase I (PGFS I) and prostaglandin F synthase II (PGFS II). Prostaglandin F(2alpha) binds to its specific receptor, FP. Our previous immunohistochemical study showed the distinct localization of prostaglandin F synthases in rat spinal cord. PGFS I exists in neuronal somata and dendrites in the gray substance, and PGFS II exists in ependymal cells and tanycytes surrounding the central canal. Both enzymes are also present in endothelial cells of blood vessels in the white and gray substances of the spinal cord. In this study, we found that FP localizes in neuronal somata and dendrites but not in ependymal cells, tanycytes, or endothelial cells. Immunohistochemical analysis of serial sections showed the colocalization of FP and PGFS I. FP immunoreactivity was intense in spinal laminae I and II of the dorsal horn, a connection site of pain transmission, and was similar to that of PGFS I in neuronal elements. These findings suggest that prostaglandin F(2alpha) synthesized in the neuronal somata and dendrites exert an autocrine action there.