We have prepared antibodies against porcine liver betaine–homocysteine methyltransferase (BHMT; EC 2.1.1.5) and recently cloned cDNAs encoding the porcine and human liver enzymes. Porcine tissues were evaluated for BHMT expression by measuring catalytic activity and Western analysis. Liver and kidney were the only organs tested that had immunodetectable levels of BHMT, and these organs expressed high levels of enzyme activity. BHMT was expressed in the kidney cortex and not the medulla. Porcine pancreas, brain, heart, lung, and spleen were devoid of BHMT activity and immunodetectable protein. Human tissues were tested for BHMT expression by Northern analysis. Human liver and kidney were the only organs tested that expressed BHMT mRNA. Human pancreas, brain, heart, skeletal muscle, spleen, and placenta were devoid of BHMT mRNA. The human BHMT gene has been mapped to chromosome 5q13.1-q15.

Chronic diseases develop in susceptible individuals following exposure to environmental conditions including high fat diets. Inbred strains of mice differing in susceptibility to atherosclerosis, diabetes, obesity and certain cancers are models for understanding the genetic basis and molecular mechanisms whereby diet influences these polygenic and multifactorial disorders. Expression sequence tags (EST) and disease quantitative trait loci (QTL) are also being identified with these strains. Reported here are comparisons of food intake, growth, nonfasting serum lipids and expression of mRNA for hepatic apolipoprotein E (ApoE), hepatic stearoyl CoA desaturase (Scd1) and heart lipoprotein lipase (Lpl) in a 2 x 2 x 2 design with C57BL/6J and BALB/cByJ mice fed semipurified diets with 4 or 20% saturated (coconut) or unsaturated (corn) oils for 4 mo. Histological studies of aortas and coronary arteries are also reported for these animals. After 4 mo, BALB/cByJ mice were significantly heavier and had significantly higher total serum cholesterol, HDL cholesterol and triglyceride concentrations in the fed state than C57BL/6J mice. Efficiency of utilizing dietary energy did not differ consistently between strains. Oil level affected serum total cholesterol, triglycerides and HDL cholesterol, which were significantly greater in mice fed high fat diets. Lpl and ApoE mRNA expression levels were not significantly affected by mouse strain, oil source or oil level. Scd1 mRNA expression, however, was significantly higher in C57BL/6J than in BALB/cByJ mice and was lower in all mice fed 20% compared with those fed 4% fat diets. Genes regulated differently by diet among strains with distinct susceptibility to diet-influenced disease may be associated with molecular pathways contributing to incidence or severity.

Genomic sequences encoding murine Lfm1, whose predicted protein sequence is 96% and 98% similar to bovine and rat F1F0-ATP synthase e subunits (respectively), have been amplified from BALB/cByJ DNA, cloned, and sequenced. The 1,1-kilobase gene has 3 introns and 4 exons, and its coding sequence differs by two nucleotides compared to the previously published BALB/cHnn Lfm1 cDNA sequence. A PstI restriction site polymorphism in intron 2 between C57BL/6J and Mus spretus was used to map this gene to Chromosome 5 near D5Mit9. Related sequences mere mapped on Chromosomes 8, 11, and 2 unlinked loci on Chromosome 2 using Southern blot analyses with the 1.1-kilobase gene as probe. Previous studies from this laboratory indicated that the Lfm1/e subunit was regulated by the level of dietary fat and carbohydrate. Northern hybridization analyses demonstrated that e subunit mRNA abundance showed statistically significant differences (p < 0.025) between hearts of BALB/c mice fed 3% and those fed 20% corn oil for 2 weeks and in liver (p < 0.05) from the same animals. Significant differences were also observed in hepatic and heart mRNA expression at different times after eating in animals subjected to a fast/refeed regimen. The implications of the high degree of sequence similarity to the e subunit for rat and bovine F1F0-ATP synthase and its regulation by diet are discussed.

This study describes and quantifies a mechanical skin damage resulting from the redistribution of stresses caused by drilling a well in a permeable material. The damage is localized within a cylindrical ring centered at the well axis. The stresses (and related positive skins) increase with depth, angle of inclination, and production time. They rapidly fade with lateral radial distance. The damage becomes insignificant at a distance greater than four times the borehole radius (or elastic/plastic interface radius).

Databases for genes expressed in humans or cell cultures are being developed as a part of the Human Genome Project. Because genomes respond to nutritional and other environmental variables, quantitative analyses of mRNA abundance under defined nutritional and physiological states are required to understand normal metabolism and to clarify differences between normal and disease phenotypes. Reported here are comparisons of food intake, growthp5erum lipids and expression of mRNA for hepatic stearoyl CoA desaturase (Scd1) and heart lipoprotein lipase (Lpl) in female BALB/cHnn mice following food deprivation and refeeding at the end of 2 wk of feeding semipurified diets with 3, 10 or 20% corn oils. Body weights and utilization of dietary energy were similar for mice fed all three diets. There were no differences in serum lipid concentrations associated with the level of dietary fat during subsequent food deprivation and refeeding, but significant differences in serum triglycerides and total serum cholesterol were observed between food-deprived and fed mice. Heart lipoprotein lipase and hepatic Scd1 mRNA expression levels were affected significantly by concentration of corn oil and by time after eating. These and other studies examining gene regulation by dietary variables and nutrient availability are discussed in relation to development of diet-regulated gene databases for laboratory animals fed semipurified diets.

Summary This paper describes a one-step fracturing/gravel-pack (frac-and-pack) completion procedure conducted on the BP Exploration Amberjack platform beginning in early 1992. This platform is 35 miles southwest of Venice, LA. The first four completions on this platform had an average positive skin values of 21. The goal of the frac-and-pack procedure was to reduce these skins to nearly zero. In total, 24 frac-and-pack operations were performed. Details of the fracture design, prefracture testing, fracture design and execution, and production response and a continuing optimization program are discussed. The fractures were performed with the screens in place with the gravel pack after the fracturing operation. The treatments were designed for the tip screenout technique1,2 to create wide fractures and to provide proppant loadings exceeding 8 lbm/ft2. This paper presents the trend of the declining skin values, along with a discussion of time-dependent skins. The changes in fluids, breakers, and proppants are also presented. The average skin on 14 frac-and-pack completions was 5.3. The average skin on the final eight completions was 0.2.