Cloning and prokaryotic expression of the porcine lipasin gene.

Lipasin has recently been demonstrated to be involved in lipid metabolism. In this study, two specific primers were used to amplify the lipasin open reading frame from porcine liver tissue. The polymerase chain reaction product was cloned to a pGEM®-T Easy Vector, digested by SalI and NotI, and sequenced. The lipasin fragment was then cloned to a pET21(b) vector and digested by the same restriction enzyme. The recombinant plasmid was transferred to Escherichia coli (BL21), and the lipasin protein was induced with isopropyl-ß-D-thiogalactopyranoside. The protein obtained was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. A pET-lipasin prokaryotic recombinant expression vector was successfully constructed, and a 25.2-kDa protein was obtained. This study provides a basis for further research on the biological function of porcine lipasin.

Identification of single nucleotide polymorphisms in the ASB15 gene and their associations with chicken growth and carcass traits.

ASB15 is a member of the ankyrin repeat and suppressor of cytokine signaling box family, and is predominantly expressed in skeletal muscle. In the present study, an F2 resource population of Gushi chickens crossed with Anka broilers was used to investigate the genetic effects of the chicken ASB15 gene. Two single nucleotide polymorphisms (SNPs) (rs315759231 A>G and rs312619270 T>C) were identified in exon 7 of the ASB15 gene using forced chain reaction-restriction fragment length polymorphism and DNA sequencing. One was a missense SNP (rs315759231 A>G) and the other was a synonymous SNP (rs312619270 T>C). The rs315759231 A>G polymorphism was significantly associated with body weight at birth, 12-week body slanting length, semi-evisceration weight, evisceration weight, leg muscle weight, and carcass weight (P < 0.05). The rs312619270 T>C polymorphism was significantly associated with body weight at birth, 4, 8, and 12-week body weight, 8-week shank length, 12-week breast bone length, 8 and 12-week body slanting length, breast muscle weight, and carcass weight (P < 0.05). Our results suggest that the ASB15 gene profoundly affects chicken growth and carcass traits.

Molecular cloning and tissue distribution profiles of the chicken R-spondin1 gene.

Rspo1 belongs to the Rspo family, which is composed of 4 members (Rspo1-4) that share 40 to 60% sequence homology and similar domain organizations, and regulate the WNT signaling pathway via a common mechanism. Rspo1 plays a key role in vertebrate development and is an effective mitogenic factor of gastrointestinal epithelial cells. We report the cloning of chicken Rspo1 and its gene expression distribution among tissues. It contained an open reading frame of 783 bp encoding a protein of 260 amino acids, and its molecular weight was predicted to be 28.80 kDa. Reverse transcription-polymerase chain reaction-based gene expression analysis indicated that chicken Rspo1 was highly expressed in the stomach muscle tissue, but was expressed at low levels in the lung, brain, jejunum, cecum, ileum, spleen, pancreas, kidney, and glandular stomach. These results suggest that Rspo1 plays a major role in muscular immune protection.

Attenuated mRNA expression of lipid metabolism genes in primary hepatocytes following lipopolysaccharide treatment in dairy cows.

The influence of ruminal acidosis on ruminal microbiology and metabolite production has received considerable attention, but little is known regarding the systemic manifestations that arise from ruminal acidosis. Lipopolysaccharide (LPS) is released in the gastrointestinal tract upon ingestion of high-grain or high-fat diets, and it has been implicated in the etiology of multiple energy- and lipid-related metabolic disturbances in ruminants. The liver plays a crucial role in the acute phase response to intruding pathogens. The effect of blood LPS in subacute ruminal acidosis on lipid metabolism in the liver has not been established. In this study, cell cultures were photographed using an inverted microscope. We observed that hepatocytes changed their morphologies from irregular triangle to circular (contraction) shapes; the number of contracted cells increased with the increasing LPS doses. This suggests that LPS can promote cell contraction and take off the wall, ultimately leading to cell apoptosis. With changes in LPS exposure, hepatocyte number also changes. We explored lipid metabolism in the liver using quantitative reverse transcription-polymerase chain reaction to detect the expression of key lipid metabolism enzymes in hepatocytes. We found that Toll-like receptor 4 signaling pathway mediated by LPS could attenuate mRNA expression of fatty acid synthesis genes and increase the expression of fatty acid transport genes in primary hepatocytes following LPS treatment in dairy cows.

SU-D-213CD-03: Live Video-Guided Volumetric Tracking of Respiration Motion.

PURPOSE: To introduce video surface imaging guidance in synchronization with 4D cone-beam CT (CBCT) scans, and in combination with respiration- gated or target-tracked dose delivery to treat mobile tumors, without collaterally damaging nearby critical structures. METHODS: The approach uses the concept that the integral of balanced forces over the moving surfaces is directly proportional to the lung volume changes. The respiratory motions, representing the lung volume variations, were measured with the dynamic volume under the moving surfaces of the thorax and abdomen. Sequential surface images on several patients and volunteers were acquired for the feasibility study. Respiratory motions were repeatedly measured on volunteers undertaking a quiet (normal) or a forced (deep) breath. The dynamic volume under the moving surfaces were robustly fitted with a linear trend and a trigonometric wave function that was compared with the fitted curves for target moving trajectories derived from forty 4D-CBCT scans. RESULTS: A large chest wall superior-outward movement was the unique characteristic of a forced breath that had doubled the volume variations and elongated the respiration period from quiet breath of ∼4 seconds to >6 seconds. Under a quiet breath, target motion trajectories could be easily described by single sine functions that were consistent with dynamic surface volume modeling except for having different motion amplitudes. The accuracy in synchronization of the real-time surface motion with respiration motion was within the measurement uncertainty of ∼2 mm. CONCLUSIONS: The analytical results with a hypothetical single sine platform allow us to accurately predict internal target motion with use of real-time video images. Synchronization of dynamic volume with respiratory motion appears applicable for association of 4D medical imaging with 4D videoimaging.

Isolation and characterization of 16 novel microsatellite loci in two inbred strains of the Chinese hamster (Cricetulus griseus).

The Shanyi inbred A and E strains of the Chinese hamster are widely used in biomedical research, but detailed genetic characterization has been lacking. We developed microsatellite markers that could be used for genetic diversity analysis and linkage map construction. We isolated and characterized 16 novel microsatellite loci from a microsatellite-enriched genomic DNA library. These loci were genotyped in 48 animals from the two strains, and the polymorphic information content was determined. In the Shanyi A and E populations, 14 and 15 loci were found to be polymorphic, respectively, with polymorphic information content ranging from 0.1393 to 0.8082 and from 0.1109 to 0.7397, respectively. A total of 115 alleles were found for the 16 microsatellite loci in the two populations; the mean observed heterozygosity (H(O)) was 0.5191 and 0.4333 for the A and E populations, respectively, indicating marked genetic variation within the two populations. Correspondingly, the F(ST) values ranged from 0.002 to 0.9253, with an overall mean of 0.1935, indicating significant genetic difference between the two strains. The population differentiation levels were substantiated by Nei's genetic distance and full Bayesian analyses computed with STRUCTURE. Despite the genetic diversity and differentiation within and between the two inbred populations, the 48 individuals were correctly allocated into their original populations with high statistical confidence based on these 16 microsatellite loci. These novel microsatellite loci should be useful genetic markers for these two Chinese hamster inbred strains.

Development of a program of behavior modification directed to the rehabilitation of drug-dependent patients: treatment and follow-up of 223 cases.

The study reports treatment and follow-up of compulsive drug-consuming patients (mainly of coca paste). The program used was based on a behavioral cognitive and instructional model. The traditional functional analysis was modified to include the therapeutical work in seven behavioral areas: (1) drug use; (2) behavior during free time; (3) behavior at work; (4) social behavior; (5) self- and environmental management behaviors; (6) problem solving and decision-making behaviors; (7) recognition, evaluation, and modification of irrational beliefs. For each area objectives, therapeutical procedures, control and evaluation methods, and termination criteria were determined. Patients engaged in a multiple activity program and received individual and group therapy. Out of 223 male patients, 130 were discharged (that is, they fulfilled all the conditions stated by the program) and 93 patients abandoned treatment. For evaluation purposes a test was used to determine the accomplishment of the behavioral objectives. Follow-up interviews after 6 to 72 months showed that although 24 patients relapsed to drug use, 106 (81.48%) of the patients who had finished the program restrained from using drugs and obtained high scores in all seven behavioral areas.