Use of bovine EST data and human genomic sequences to map 100 gene-specific bovine markers

A system to use bovine EST data in conjunction with human genomic sequence to improve the bovine linkage map over the entire genome or on specific chromosomes was evaluated. Bovine EST sequence was used to provide primer sequences corresponding to bovine genes, while human genomic sequence directed primer design to flank introns and produce amplicons of appropriate size for efficient direct sequencing. The sequence tagged sites (STS) produced in this way from the four sires of the MARC reference families were examined for single nucleotide polymorphisms (SNPs) that could be used to map the corresponding genes. With this approach, along with a primer/extension mass spectrometry SNP genotyping assay, 100 ESTs were placed on the bovine genetic linkage map. The first 70 were chosen at random from bovine EST–human genomic comparisons. An additional 30 ESTs were successfully mapped to bovine Chromosome 19 (BTA19), and comparison of the resulting BTA19 map to the position of the corresponding human orthologs on the HSA17 draft sequences revealed differences in the spacing and order of genes. Over 80% of successful amplicons contained SNPs, indicating that this is an efficient approach to generating EST-associated genetic markers. We have demonstrated the feasibility of constructing a linkage map based on SNPs associated with ESTs and the plausibility of utilizing EST, comparative mapping information, and human sequence data to target regions of the bovine genome for SNP marker development.     

Marine amoebae from waters of northwest Spain, with comments on a potentially pathogenic euryhaline species

Of 17 species of free-living amoebae identified in various samples of salt water, only 1, Acanthamoeba polyphaga, is known to be a potential pathogen. While no deaths occurred when laboratory animals were inoculated with A. polyphaga to test for pathogenicity, the protozoa were present in the brain, liver and lungs of some but not all of the animals.     

An assessment of the role of domain F and pest sequences in estrogen receptor half-life and bioactivity

The estrogen receptor (ER) is a rapidly turning over protein, with a half-life of ca. 3–4 h in estrogen target cells. Sequence analysis of the human ER reveals a putative PEST sequence, sequences rich in proline (P), glutamic acid (E), serine (S) and threonine (T), in the carboxy-terminal F domain of the protein. Since PEST sequences have been implicated in the rapid turnover of some proteins, we have used site-directed mutagenesis to investigate the role of the F region containing PEST residues in the stability and bioactivity of the receptor. A truncated form of ER lacking the last 41 amino acids of the protein and encompassing the PEST sequences (amino acids 555 to 567) was made by mutagenesis of the ER cDNA. Pulse-chase experiments, involving immunoprecipitation of [³⁵S]methionine/[³⁵]Scysteine labeled receptors or of receptors covalently labeled with tamoxifen aziridine followed by gel electrophoresis, were used to determine the half-life of the wild-type and truncated ERs. These experiments showed that the turnover rate of the receptors expressed in Chinese hamster ovary and monkey kidney (COS-1) cells was 3 to 5 h and that elimination of the PEST residues did not have a significant effect on the degradation rate of the protein. Moreover, deletion of the last 41 amino acids (F domain) of the ER did not affect transactivation ability, ligand binding affinity, or the phosphorylation pattern of the receptor. Therefore, the role of domain F in ER function remains unclear, but it is not a determinant of the relatively rapid rate of ER turnover in cells.     

FBXO44-Mediated Degradation of RGS2 Protein Uniquely Depends on a Cullin 4B/DDB1 Complex

The ubiquitin-proteasome system for protein degradation plays a major role in regulating cell function and many signaling proteins are tightly controlled by this mechanism. Among these, Regulator of G Protein Signaling 2 (RGS2) is a target for rapid proteasomal degradation, however, the specific enzymes involved are not known. Using a genomic siRNA screening approach, we identified a novel E3 ligase complex containing cullin 4B (CUL4B), DNA damage binding protein 1 (DDB1) and F-box protein 44 (FBXO44) that mediates RGS2 protein degradation. While the more typical F-box partners CUL1 and Skp1 can bind FBXO44, that E3 ligase complex does not bind RGS2 and is not involved in RGS2 degradation. These observations define an unexpected DDB1/CUL4B-containing FBXO44 E3 ligase complex. Pharmacological targeting of this mechanism provides a novel therapeutic approach to hypertension, anxiety, and other diseases associated with RGS2 dysregulation.     

Lactic acid bacteria: hydrophobicity and strength of attachment to meat surfaces

The hydrophobicity and strength of a ttachment of several lactic acid bacteria with antimicrobial activity were studied. Hydrophobicity was determined by bacterial adherence to hydrocarbons (BATH; octane or xylene), adhesion to nitrocellulose filters (NCF), salt aggregation test (SAT) and adherence to phenyl–Sepharose beads (PSB). The relative hydrophobicity of lactic acid bacteria depended markedly on the method used. No correlation between either SAT or BATH (octane) and strength of attachment (Sr value) existed. However, a significant relationship between strength of attachment and BATH (xylene), NCF and PSB, respectively, was observed, showing the highest correlation coefficient ( r = 0·778) for BATH (xylene).     

Ghrelin Causes a Decline in GABA Release by Reducing Fatty Acid Oxidation in Cortex

Lipid metabolism, specifically fatty acid oxidation (FAO) mediated by carnitine palmitoyltransferase (CPT) 1A, has been described to be an important actor of ghrelin action in hypothalamus. However, it is not known whether CPT1A and FAO mediate the effect of ghrelin on the cortex. Here, we show that ghrelin produces a differential effect on CPT1 activity and γ-aminobutyric acid (GABA) metabolism in the hypothalamus and cortex of mice. In the hypothalamus, ghrelin enhances CPT1A activity while GABA transaminase (GABAT) activity, a key enzyme in GABA shunt metabolism, is unaltered. However, in cortex CPT1A activity and GABAT activity are reduced after ghrelin treatment. Furthermore, in primary cortical neurons, ghrelin reduces GABA release through a CPT1A reduction. By using CPT1A floxed mice, we have observed that genetic ablation of CPT1A recapitulates the effect of ghrelin on GABA release in cortical neurons, inducing reductions in mitochondrial oxygen consumption, cell content of citrate and α-ketoglutarate, and GABA shunt enzyme activity. Taken together, these observations indicate that ghrelin-induced changes in CPT1A activity modulate mitochondrial function, yielding changes in GABA metabolism. This evidence suggests that the action of ghrelin on GABA release is region specific within the brain, providing a basis for differential effects of ghrelin in the central nervous system.     

Ethnobotany and domestication in Xoconochtli,Stenocereus stellatus (Cactaceae), in the Tehuacán Valley and La Mixteca Baja, México

Ethnobotanical information is presented on use, management, folk nomenclature and classification of the “xoconochtli” (Stenocereus stellatus) as well as on the role of this plant in subsistence of the Nahua, Mixtec and Popoloca peoples from the Tehuacán Valley and La Mixteca Baja in Central Mexico. Among all three groups, S. stellatus was used for various purposes but mainly for its edible fruits. Different variants of this species were distinguished, named and classified by indigenous people according to characteristics of the fruit; particularly size, color and flavor of the pulp, spininess and thickness of the peel. Wild plants characteristically had small red sour fruits with many spines and thick peel, while individuals selected for cultivation usually differed in one or more of these characters. Three general forms of interaction between people and this species were found: 1) gathering of useful products from the wild; 2) management of wild populations in situ which involves the sparing and enhancing of individuals with more desirable characteristics and the removal of others during clearance of the land for agriculture; and 3) cultivation, mainly in home gardens, by propagation of vegetative parts from desirable individuals. Fruit yields per individual and per population were measured and compared in wild, managed in situ and cultivated populations from the Tehuacán Valley and La Mixteca Baja. Wild and cultivated individuals from La Mixteca Baja yielded more than wild and cultivated individuals from the Tehuacán Valley. Within each region, fruit yields were similar in wild and managed in situ populations but significantly larger in cultivated populations. Forms of management of this plant species are discussed in terms of availability of products and demand for them in the local economy.     

Maximal aerobic capacity across age in healthy Hispanic and Caucasian women.

We tested the hypothesis that the age-related decline in maximal aerobic capacity, as measured by maximal oxygen uptake (VO(2 max)), is greater in Hispanic than in Caucasian women. We studied 146 healthy sedentary women aged 20-75 yr: 53 Hispanic (primarily of Mexican descent) and 93 Caucasian (non-Hispanic white). The groups did not differ in mean age, body mass, percent body fat, estimated physical activity-related energy expenditure, or education-based socioeconomic status (SES). During maximal exercise, respiratory exchange ratio, rating of perceived exertion, and percent predicted maximal heart rate were similar across age and ethnicity, suggesting equivalent maximum voluntary efforts in all subjects. VO(2 max) (ml x kg(-1) x min(-1)) was inversely related to age (P < 0.01) in Caucasian (r =-0.68) and Hispanic (r = -0.61) women. The absolute rate of decline in VO(2 max) with age was the same in the two groups (-0.31 ml x kg(-1) x min(-1) x yr(-1)). The relative rate of decline (% from age 25 yr) also was similar in the Caucasian (-9.0%) and Hispanic (-9.2%) women. When subjects of all ages were pooled, mean levels of VO(2 max) were similar in the two groups (approximately 28 ml x kg(-1) x min(-1)). These results, the first to our knowledge in Hispanics, indicate that mean levels of VO(2 max), as well as the rate of decline in VO(2 max) with age, are similar in healthy sedentary Hispanic and Caucasian women of similar SES. Thus it does not appear that Hispanic ethnicity per se modulates maximal aerobic capacity in this population.     

A comparative study of phagocytic activity and lymphoproliferative response in five varieties of tilapia Oreochromis spp.

The immunological status of three native species of tilapia Oreochromis spp. and two hybrids were compared, showing a greater potential of the hybrid strain O. niloticus Rocky Mountain for aquaculture and a better capacity to resist the stressful conditions of a fish farm.