Regulatory gene network from a genome-wide association study for sow lifetime productivity traits.

Among swine reproductive traits, sow lifetime productivity (SLP) is considered a profitable trait in commercial pig farming. Notably, longevity and efficiency in SLP can be adopted as the key phenotype representing SLP. In this study, we conducted a co-association network analysis using results from a genome-wide association study for SLP-related traits. A total of 656 purebred Landrace female pigs were genotyped using a 60K SNP array. Significantly associated SNPs identified from the GWAS were annotated for the specific genes. Then, we constructed an association weight matrix to build a network based on the co-associations between the genes and 10 SLP traits. The entire network consisted of 495 nodes and 37 755 significant edges. We identified three key regulatory transcription factors: STAT2 (signal transducer and activator of transcription 2), MYF6 (myogenic factor 6) and TFCP2L1 (transcription factor CP2 like 1). The network revealed that the STAT2 and MYF6 regulatory modules cooperate with each other and specifically influence the longevity and efficiency of sows, whereas the TFCP2L1 family specifically affects the improvement of litter size.

Genomic characterization of the porcine CRTC3 and the effects of a non-synonymous mutation p.V515F on lean meat production and belly fat.

cAMP-responsive element-binding protein (CREB)-regulated transcriptional coactivator 3 (CRTC3) is well known to be related to obesity in humans and mice. However, the effects of CRTC3 have not been studied in pigs. Here, we characterized the structure of the porcine CRTC3 gene and identified single nucleotide polymorphisms (SNPs) in its coding region. Moreover, mRNA expression profiles of CRTC3 in muscle and fat tissues were examined. Of the 40 identified SNPs, the p.V515F mutation, located on exon 16, was genotyped in 368 Yorkshire pigs. The p.V515F mutation was significantly associated with lean meat production ability, including reduced back fat thickness (P=0.0317) and loin eye area (P=0.0174). Moreover, the SNP was significantly associated with differences in intermuscular fat (P=0.0092), total muscle area in the belly (P=0.0108), and total fat percentage in the belly (P=0.0298). Taken together, our results suggest that the p.V515F mutation affects to lean meat production ability and amount of belly fat.

Pharmacokinetic indices for cefovecin after single-dose administration to adult sea otters (Enhydra lutris).

Seven sea otters received a single subcutaneous dose of cefovecin at 8 mg/kg body weight. Plasma samples were collected at predetermined time points and assayed for total cefovecin concentrations using ultra-performance liquid chromatography and tandem mass spectrometry. The mean (±SD) noncompartmental pharmacokinetic indices were as follows: CMax (obs) 70.6 ± 14.6 µg/mL, TMax (obs) 2.9 ± 1.5 h, elimination rate constant (kel ) 0.017 ± 0.002/h, elimination half-life (t1/2kel) 41.6 ± 4.7 h, area under the plasma concentration-vs.-time curve to last sample (AUClast) 3438.7 ± 437.7 h·µg/mL and AUC extrapolated to infinity (AUC0→∞ ) 3447.8 ± 439.0 h·µg/mL. The minimum inhibitory concentrations (MIC) for select isolates were determined and used to suggest possible dosing intervals of 10 days, 5 days, and 2.5 days for gram-positive, gram-negative, and Vibrio parahaemolyticus bacterial species, respectively. This study found a single subcutaneous dose of cefovecin sodium in sea otters to be clinically safe and a viable option for long-acting antimicrobial therapy.

Comparison of Strain in GaN-Based Blue Light-Emitting Diode Grown on Silicon(111) and Sapphire Substrates.

We compare the strain states and device performances of GaN-based blue light-emitting diodes (LEDs) grown on Si(111) and sapphire substrates. The strain characteristics are investigated using micro-Raman spectroscopy and high-resolution transmission electron microscopy. These analyses reveal that GaN layer grown on Si has a residual tensile strain in contrast to a compressive strain for GaN on sapphire, and quantum wells (QWs) on GaN/Si experience reduced lattice mismatch than those of GaN/sapphire. When external quantum efficiencies of LED on sapphire and Si substrates are compared, the LED on Si shows better efficiency droop characteristics and this is attributed to a decrease in piezo-electric field strength in InGaN/GaN layers owing to reduced lattice mismatch.

Estimation of pork quality traits using exsanguination blood and postmortem muscle metabolites.

The current study was designed to estimate the pork quality traits using metabolites from exsanguination blood and postmortem muscle simultaneously under the Korean standard pre- and post-slaughter conditions. A total of 111 Yorkshire (pure breed and castrated male) pigs were evaluated under the Korean standard conditions. Measurements were taken of the levels of blood glucose and lactate at exsanguination, and muscle glycogen and lactate content at 45 min and 24 h postmortem. Certain pork quality traits were also evaluated. Correlation analysis and multiple regression analysis including stepwise regression were performed. Exsanguination blood glucose and lactate levels were positively correlated with each other, negatively related to postmortem muscle glycogen content and positively associated with postmortem muscle lactate content. A rapid and extended postmortem glycolysis was associated with high levels of blood glucose and lactate, with high muscle lactate content, and with low muscle glycogen content during postmortem. In addition, these were also correlated with paler meat color and reduced water holding capacity. The results of multiple regression analyses also showed that metabolites in exsanguination blood and postmortem muscle explained variations in pork quality traits. Especially, levels of blood glucose and lactate and content of muscle glycogen at early postmortem were significantly associated with an elevated early glycolytic rate. Furthermore, muscle lactate content at 24 h postmortem alone accounted for a considerable portion of the variation in pork quality traits. Based on these results, the current study confirmed that the main factor influencing pork quality traits is the ultimate lactate content in muscle via postmortem glycolysis, and that levels of blood glucose and lactate at exsanguination and contents of muscle glycogen and lactate at postmortem can explain a large portion of the variation in pork quality even under the standard slaughter conditions.

Effects of hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 polymorphisms on fat androstenone level and gene expression in Duroc pigs.

A high level of androstenone in porcine adipose tissue is a major factor contributing to boar taint. Porcine hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (3ß-HSD, also known as HSD3B1) plays a key role in the hepatic metabolism that catalyzes androstenone to ß-androstenol. Therefore, 3ß-HSD is a candidate gene for boar taint. This study aimed to investigate functional 3ß-HSD polymorphisms in Duroc pigs. We found eight single nucleotide polymorphisms (SNPs) in the full-length porcine 3ß-HSD. Four of the SNPs had restriction enzyme sites, and we genotyped them in 147 uncastrated male Duroc pigs using a polymerase chain reaction-restriction fragment length polymorphism method. Pigs with the GG genotype at the g.165262G>A locus (SNP5) had significantly lower androstenone levels than did those with other genotypes (P = 0.030). SNP5 also was associated with differences in 3ß-HSD mRNA levels: pigs with the GG genotype had higher levels than those with other genotypes (P = 0.019). The SNP5 polymorphism could affect the hepatic catabolism of androstenone and consequently impact androstenone accumulation in the adipose tissue. Therefore, SNP5 in the 3ß-HSD of Duroc pigs could be a useful selective marker for decreasing boar taint.

Avian polyomavirus identified in a nestling Gouldian finch (Erythrura gouldiae) in New Zealand.

Abstract CASE HISTORY: Four 4-5-month-old nestlings and one adult in a commercial aviary of 53 Gouldian finches (Erythrura gouldiae) died over a 2-week period in July 2000. PATHOLOGICAL FINDINGS: One nestling was necropsied and showed bronze-tinged skeletal muscles, a swollen liver with haemorrhagic margins and numerous haemorrhages on serosal surfaces. The histological lesions included multifocal hepatic necrosis and haemorrhage associated with the presence of large clear or basophilic intranuclear inclusions in hepatocytes and Kupffer cells suggestive of avian polyomavirus infection. Similar inclusion bodies were present in splenic histiocytes. MOLECULAR BIOLOGY: DNA was subsequently extracted from archived portions of liver, spleen, gizzard, heart, lung and kidney. A broad spectrum nested PCR was used to detect polyomavirus which sequence analysis confirmed as finch polyomavirus. DIAGNOSIS: Avian polyomavirus. CLINICAL RELEVANCE: Avian virus infections such as polyomavirus should be suspected in cases of sudden death in nestlings, particularly in susceptible species such as psittacine and passerine birds. The archiving of tissues from unconfirmed disease outbreaks provides a valuable resource for retrospective investigations.

Effects of variation in porcine MYOD1 gene on muscle fiber characteristics, lean meat production, and meat quality traits.

Three single nucleotide polymorphisms (SNPs) in the porcine MYOD1 gene were used for association analysis and haplotype construction to evaluate the effects of their substitution. Four hundred and three pigs of Yorkshire and Berkshire breeds were used. The mRNA expression levels of MYOD1 were examined. The g.489C>T and g.1264C>A SNPs were significantly associated with several muscle fiber characteristics, the loin eye area, and lightness. Particularly, animals having hetero-genotypes of both sites showed good performance both in lean meat production and meat quality traits. The results of haplotype substitution were similar to the associations of individual SNPs. Moreover, the 2 SNPs had significant effects on mRNA expression. Therefore, the g.489C>T and g.1264C>A SNPs in MYOD1 may be meaningful DNA markers that can be used for improving important porcine economic traits.

Polymorphisms of the 5' regulatory region of the porcine PPARGC1A gene and the effects on muscle fiber characteristics and meat quality.

The purpose of this study was to determine the structure of the porcine PPARGC1A 5' upstream region, and to find suitable molecular markers for improved meat quality and good lean meat production. Ten DNA polymorphisms, including 7 SNPs, 2 microsatellites, and 1 insertion or deletion were newly found in the 5' upstream region of PPARGC1A. Three SNPs that had restriction enzyme site were evaluated for associations with muscle fiber characteristics and production traits. Two hundred fifty-two pigs (Yorkshire and Landrace) were used in this analysis. The c.-2894G>A genotypes was significantly associated with muscle fiber characteristics, including the number of fiber type I and IIb composition (P < 0.05), mean cross-sectional area of fibers (P < 0.01), and fiber number per unit area (P < 0.05). The animals with the GG genotype had a higher percentage of type I fibers and a lower percentage of type IIb fibers with better meat quality [higher pH value (P < 0.05) and lower drip loss (P < 0.05)] and lean meat production [larger loin eye area (P < 0.05)]. Moreover, the mRNA expression levels of PPARGC1A among genotypes were significantly different with the highest level of GG genotype. The c.-2885G>T and c.-1402A>T sites showed similar results that had significant effects on the mean cross-sectional area (CSA; P < 0.05), fiber number per unit area (P < 0.05) and loin eye area (P < 0.01). Therefore, we suggest that the c.-2894G>A polymorphism in the 5' upstream region of the porcine PPARGC1A gene can be used as a meaningful molecular marker for simultaneous improvement of lean meat production and quality traits.

Robustness of QTLs across germplasm pools using a model quantitative trait.

Knowledge of the inheritance of C-glycosyl flavone synthesis in maize (Zea mays L.) silk tissues has been acquired through detailed genetic studies involving primarily germplasm from the Corn Belt Dent race. To test the robustness of this genetic knowledge, we examined C-glycosyl flavone synthesis in a genetically distinct germplasm pool, popcorn. C-glycosyl flavone profiles and levels and the involvement of three specific genes/quantitative trait loci (p, pr1, and sm1) in C-glycosyl flavone synthesis were examined in popcorn germplasm representing at least two races and various diverse geographic regions. Twenty-four inbred lines and 23 hybrids involving these inbred lines and inbred line R17 were characterized for their flavone profiles and levels in silk tissues. Two F2 mapping populations were constructed to examine the involvement of p, pr1, and sm1 on C-glycosyl flavone synthesis. C-glycosyl flavone levels threefold higher than previously reported in Corn Dent Belt materials and a novel class of compounds were discovered. The gene action of sm1 was different, the functional p allele was not always dominant, and pr1 did not affect mays in synthesis. Based on this rather simplistic "model" quantitative trait, it appears that caution should be exercised when attempting to apply quantitative trait locus knowledge accumulated in one germplasm base to a germplasm base that is known to be distinctly unique.

Pharmacokinetics of oral doxycycline and concentrations in body fluids and bronchoalveolar cells of foals.

The objective of this study was to determine the disposition of orally administered doxycycline in foals. Six healthy 4- to 8-week-old foals were used. Doxycycline was administered to each foal via the intragastric (IG) route at dosages of 10 and 20 mg/kg, in a cross-over design. After the first 10 mg/kg dose, five additional doses were administered at 12-h intervals. A microbiological assay was used to measure doxycycline activity in serum, urine, peritoneal fluid, synovial fluid, cerebrospinal (CSF), pulmonary epithelial lining fluid (PELF), and bronchoalveolar (BAL) cells. Following administration at 10 mg/kg, mean+/-SD time to peak serum doxycycline activity (tmax) was 3.0+/-1.2 h, maximum serum activity (Cmax) was 2.54+/-0.27 microg/mL, and terminal half-life (t1/2) was 8.5+/-2.8 h. Administration at a dose of 20 mg/kg resulted in a significantly longer tmax (5.5+/-1.8 h) as well as a tendency toward higher Cmax (2.89+/-0.33 microg/mL) and longer t1/2 (11.9+/-2.6 h). After multiple IG doses, doxycycline activity in CSF was significantly lower than concurrent serum activity, whereas peritoneal fluid, synovial fluid, and BAL cell doxycycline activity was similar to concurrent serum activity. Doxycycline activity in urine and PELF was significantly higher than that found at other sites. Oral administration at a dosage of 10 mg/kg every 12 h would maintain serum, PELF, and BAL cell activity above the minimum inhibitory concentrations of Rhodococcus equi, beta-hemolytic streptococci, and other susceptible bacterial pathogens for the entire dosing interval.

Pulmonary disposition of erythromycin, azithromycin, and clarithromycin in foals.

The objectives of the present study were to determine and compare the pulmonary disposition of azithromycin, clarithromycin, and erythromycin in foals. A single dose (10 mg/kg) of azithromycin, clarithromycin, or erythromycin was administered intragastrically to six healthy 1- to 3-month-old foals using an orthogonal design. Activity of the drugs was measured in serum, pulmonary epithelial lining fluid (PELF), and bronchoalveolar lavage (BAL) cells by use of a microbiologic assay. Peak drug activity in PELF was significantly higher in foals treated with clarithromycin (48.96+/-13.26 microg/mL) than in foals treated with azithromycin (10.00+/-7.46 microg/mL). Quantifiable erythromycin activity in PELF was only found in two of six foals. Peak drug activity in BAL cells was not significantly different between azithromycin (49.92+/-26.94 microg/mL) and clarithromycin (74.20+/-45.80 microg/mL) but activity for both drugs was significantly higher than that of erythromycin (1.02+/-1.11 microg/mL). Terminal half-life of azithromycin in serum (25.7+/-15.4 h), PELF (34.8+/-30.9 h), and BAL cells (54.4+/-17.5 h) was significantly longer than that of both clarithromycin and erythromycin. Peak azithromycin and clarithromycin activity was significantly higher in BAL cells, followed by PELF, and serum. In contrast, peak erythromycin activity in BAL cells was not significantly different from that of serum.

Induction of MMP-13 expression by soluble human glucocorticoid-induced tumor necrosis factor receptor in fibroblast-like synovial cells.

OBJECTIVE: We tested the hypothesis that human glucocorticoid-induced tumor necrosis factor receptor (hGITR/TR11) expressed on the surface of activated CD4(+) T cells is responsible for up-regulating the production of matrix metalloproteinase (MMP)-13 by fibroblast-like synoviocytes (FLSs). METHODS: The level of MMP-13 was measured by Western blot and reverse transcriptase polymerase chain reaction (RT-PCR). Expressions of hGITR ligand (hGITRL) on the surface of FLSs and hGITR on the surface of human CD4(+) T cells were analyzed by flow cytometry and RT-PCR. Neutralizing antibodies (Abs) were used to block hGITRL and hGITR on the surface of FLSs and human CD4(+) T cells, respectively. Human CD4(+) T cells were cocultured with FLSs to facilitate interaction between hGITR on CD4(+) T cells and hGITRL on FLSs. RESULTS: Soluble hGITR (shGITR) stimulated FLSs to produce MMP-13, and blockade of hGITRL reduced this effect. Direct contact between activated CD4(+) T and FLSs also induced the production of MMP-13, and neutralization of hGITR on activated CD4(+) T cells during coculture decreased the amount of MMP-13 produced by FLSs. CONCLUSION: shGITR stimulated FLSs to produce MMP-13 via a signal through hGITRL. Direct contact between activated CD4(+) T cells and FLSs facilitated hGITR-hGITRL interaction, and resulted in inducing MMP-13. This effect may increase tissue destruction in chronic inflammation such as rheumatoid arthritis (RA).

Wheat curl mite (Acari: Eriophyidae) dispersal and its relationship with kernel red streaking in maize.

Wheat curl mites, Aceria tosichella Keifer, dispersing from wheat (Triticum spp.) to nearby corn (Zea mays L.) fields play a role in the development of kernel red streaking in corn. These studies were undertaken to verify the relationship of wheat curl mite to kernel red streaking, to determine whether wheat is the main source of curl mites dispersing into corn and to determine whether planting corn in temporal or spatial isolation of wheat is a valid management strategy. These studies were conducted on farm fields using sticky traps to monitor mites, followed by sampling mature grain for kernel streaking in southwestern Ontario from 1999 to 2002. The dominant source mites were winter wheat. Mite dispersal occurred during the first 3 wk of winter wheat maturation after the wheat had reached Zadoks stage 87. Mite dispersal corresponded to prevailing winds in the area with the lowest number of mites and the lowest severity of kernel red streaking occurring 60 m from wheat fields planted to the north, south, and east of cornfields and 90 m from wheat fields planted to the west of cornfields. The severity of kernel red streaking was positively correlated with the density of wheat curl mites in corn; however, the correlation was weak and kernel red streaking was still high in many cornfields when few or no mites were present. These findings suggest that wheat curl mite migration into corn is not entirely predictive of the incidence and severity of kernel red streaking.

Blockade of 4-1BB (CD137)/4-1BB ligand interactions increases allograft survival.

We investigated the role of 4-1BB, a T cell co-stimulatory molecule, in alloimmune responses. In vivo mixed lymphocyte reactions showed that 4-1BB was preferentially expressed on actively dividing CD4(+) and CD8(+) T cells. Furthermore, following alloantigen challenge, the draining lymph nodes contained subpopulations of 4-1BB-expressing CD4(+) and CD8(+) T cells. 4-1BB-deficient C57BL/6 mice showed a delayed rejection of cardiac transplants mismatched for the major histocompatibility complex. Longer transplant survival was induced by blockade of 4-1BB/4-1BB ligand (4-1BBL) interactions using an anti-4-1BBL monoclonal antibody. Histological analysis showed that prolonged transplant survival in the 4-1BB-deficient and anti-4-1BBL-treated mice correlated with reduced lymphocytic infiltration and vasculitis in the donor heart tissue. Taken together, our data suggest that blockade of 4-1BB/4-1BBL interactions inhibited the expansion of alloreactive T cells and reduced CTL activity against host alloantigen, which in turn resulted in the prolongation of allograft survival. Blockade of the 4-1BB co-stimulatory pathway may be useful for preventing allograft rejection.

Cellular immunity induced by the recombinant Plasmodium falciparum malaria vaccine, RTS,S/AS02, in semi-immune adults in The Gambia.

Vaccination of malaria-naive humans with recombinant RTS,S/AS02, which includes the C-terminus of the circumsporozoite protein (CS), has been shown to induce strong T cell responses to both the whole protein antigen and to peptides from CS. Here we show that strong T cell responses were also observed in a semi-immune population in The Gambia, West Africa. In a Phase I study, 20 adult male volunteers, lifelong residents in a malaria-endemic region, were given three doses of RTS,S/AS02 at 0, 1 and 6 months. Responses to RTS,S, hepatitis B surface antigen and peptides from CS were tested using lymphocyte proliferation, interferon (IFN)-gamma production in microcultures, and IFN-gamma ex vivo and cultured ELISPOT, before and after vaccination. Cytotoxic responses were tested only after vaccination and none were detected. Before vaccination, the majority of the volunteers (15/20) had detectable responses in at least one of the tests. After vaccination, responses increased in all assays except cytotoxicity. The increase was most marked for proliferation; all donors responded to RTS,S after the third dose and all except one donor responded to at least one peptide after the second or third dose. There was a lack of close association of peptide responses detected by the different assays, although in microcultures IFN-gamma responses were found only when proliferative responses were high, and responses by cultured ELISPOT and proliferation were found together more frequently after vaccination. We have therefore identified several peptide-specific T cell responses induced by RTS,S/AS02 which provides a mechanism to investigate potentially protective immune responses in the field.

Linker-assisted immunoassay and liquid chromatography/mass spectrometry for the analysis of glyphosate.

A novel, sensitive, linker-assisted enzyme-linked immunosorbent assay (L'ELISA) was compared to on-line solid-phase extraction (SPE) with high-performance liquid chromatography/mass spectrometry (HPLC/MS) for the analysis of glyphosate in surface water and groundwater samples. The L'ELISA used succinic anhydride to derivatize glyphosate, which mimics the epitotic attachment of glyphosate to horseradish peroxidase hapten. Thus, L'ELISA recognized the derivatized glyphosate more effectively (detection limit of 0.1 microg/L) and with increased sensitivity (10-100 times) over conventional ELISA and showed the potential for other applications. The precision and accuracy of L'ELISA then was compared with on-line SPE/HPLC/MS, which detected glyphosate and its degradate derivatized with 9-fluorenylmethyl chloroformate using negative-ion electrospray (detection limit 0.1 microg/ L, relative standard deviation +/- 15%). Derivatization efficiency and matrix effects were minimized by adding an isotope-labeled glyphosate (2-13C15N). The accuracy of L'EUSA gave a false positive rate of 18% between 0.1 and 1.0 microg/L and a false positive rate of only 1% above 1.0 microg/L The relative standard deviation was +/- 20%. The correlation of L'ELISA and HPLC/MS for 66 surface water and groundwater samples was 0.97 with a slope of 1.28, with many detections of glyphosate and its degradate in surface water but not in groundwater.

Naturally exposed populations differ in their T1 and T2 responses to the circumsporozoite protein of Plasmodium falciparum.

T-cell responses directed against the circumsporozoite protein (CS) of Plasmodium falciparum can mediate protection against malaria. We determined the frequency of T cells reactive to different regions of the CS in the blood of donors naturally exposed to P. falciparum by examining T1 (gamma interferon [IFN-gamma] ELISPOT assay), T2 (interleukin 4 [IL-4] ELISPOT assay), and proliferative T-cell responses. The proliferative responses were weak, which confirmed previous observations. The responses to the CS in the IL-4 and IFN-gamma ELISPOT assays were also weak (<40 responding cells per 10(6) cells), much weaker than the response to the purified protein derivative of Mycobacterium tuberculosis in the same donors. Moreover, a response in one assay could not be used to predict a response in either of the other assays, suggesting that although these assays may measure different responding cells, all of the responses are weakly induced by natural exposure. Interestingly, the two different study populations used had significantly different T1 and T2 biases in their responses in the C terminus of the protein, suggesting that the extent of P. falciparum exposure can affect regulation of the immune system.

Cilia-associated respiratory bacillus infection in rats in New Zealand.

AIMS: To demonstrate cilia-associated respiratory (CAR) bacillus associated with chronic pneumonia in wild and pet rats in New Zealand. METHODS: A range of tissues from 4 rats were examined grossly and by light microscopy and affected lungs were examined by transmission electron microscopy. RESULTS: All 4 rats had moderate to severe cranioventral bronchopneumonia with bronchiectasis and large numbers of argentophilic bacteria resembling CAR bacillus, intimately associated with the bronchial epithelium. CONCLUSIONS: CAR bacillus infection should be considered as a differential diagnosis for pneumonia in rats in New Zealand.

Management of complex implant aesthetics: ensuring restorative design continuity with a comprehensive outcome-based strategy.

Since implant therapy must be dictated by prosthetic requirements, a protocol for the comprehensive and continuous integration of the restorative blueprint through the entire treatment planning and clinical execution phases is mandatory. This article demonstrates a systematic approach where the establishment of a final restorative treatment objective is incorporated into the diagnosis and subsequently integrated through every phase of clinical execution. Design continuity is ensured, and multidisciplinary cohesiveness is enhanced by providing clearly defined treatment objectives to every member of the clinical team.