The human genome and understanding of common disease: present and future technologies

The study of candidate genes over the past three decades has yielded notable successes in common-disease genetics. During this time, however, interpretation of genetic association studies has been hampered by the use of clinical cohorts of inadequate power and insufficient information on genetic variation in candidate genes. The unavailability of highthroughput and low-cost genotyping technologies has also limited the scope of complex-disease genetic studies. More recently, however, the sequencing and characterization of variation within the human genome has revolutionized genetic studies and enabled full genome-wide scans for genes associated with disease. The identification of disease-associated (causative) genes has illuminated disease mechanisms. The translation of this knowledge into direct clinical benefit in diagnosis, prognosis and therapy for an individual’s disease still remains a challenge. Received 11 September 2006; received after revision 17 December 2006; accepted 18 January 2007     

An HTLV-III peptide produced by recombinant DNA is immunoreactive with sera from patients with AIDS

Human T-cell lymphotropic retrovirus type III (HTLV-III), also called lymphadenopathy-associated virus (LAV), has been identified as the aetiological agent of acquired immune deficiency syndrome (AIDS). The sera of most patients with AIDS or AIDS-related complexes, and of asymptomatic individuals infected with HTLV-III, contain antibodies against antigens of HTLV-III. The characterization of these antibodies and their corresponding viral antigens is important not only for understanding immunity against HTLV-III and the pathology of AIDS, but also for the development of diagnostic methods and preventive vaccine for AIDS. Following the successful establishment of a long-term T-cell line permissive for HTLV-III replication, large quantities of virus have been produced, facilitating the purification of viral proteins and the development of mouse monoclonal antibodies against several viral antigens. More recently, the structure of HTLV-III proviral DNA has been elucidated. We now report the production, by genetic engineering methods, of a peptide encoded by a gene segment of HTLV-III. A 1.1-kilobase (kb) EcoRI DNA segment from an isolate of HTLV-III was inserted into a lpp and lac promoter-coupled expression vector, pIN-III-ompA. Escherichia coli transformants of this plasmid produced a peptide of relative molecular mass (Mr) 15,000 (15K) which was strongly immunoreactive with anti-HTLV-III antibodies present in sera from AIDS patients. Lysates of the clones expressing this 15K peptide inhibited the reactivity of the p31 virion protein with AIDS sera, suggesting that it is a fragment of the viral p31 protein. The peptide reacted with sera from all 20 AIDS patients but none of the 8 normal controls tested. These results suggest that the peptide may be useful for detecting anti-HTLV-III antibodies in blood samples.     

Ecology of flannelmouth sucker in the Lee's Ferry tailwater, Colorado River, Arizona

We investigated ecology of flannelmouth sucker ( Catostomus latipinnis ) from 1992 to 1997 in the 26-km Lee's Ferry reach of the Colorado River immediately below Glen Canyon Dam, Arizona. We captured by electrofishing a total of 212 fish and recaptured 52 previously tagged by others. Flannelmouth sucker were captured throughout the tailwater but tended to aggregate about 5 km of the dam, possibly reflecting blockage of historic migration routes. Catch per hour of electrofishing did not differ among years but was greater from November to February than other periods, suggesting seasonal movements of flannelmouth sucker into the tailwater: Mean lengths and weights of fish did not differ among years or seasons. Length frequency analyses also indicated there were no significantly yearly trends in proportion of catch within size classes of fish. Mean condition differed only among seasons and was greatest in February, lowest in August, coinciding respectively with pre- and post-spawning periods of flannelmouth sucker in tributary just down from Lee's Ferry. Recaptured fish migrated from initial tagging locations 1.4-23.1 km downstream from Lee's Ferry. Fifty-nine percent of recaptured fish with known initial tagging locations increased in length, and fish tagged initially as subadults or adults, respectively, grew an average of 45.9 mm and 5.5 mm per year.     

Parameters for reliable results in genetic association studies in common disease.

It is increasingly apparent that the identification of true genetic associations in common multifactorial disease will require studies comprising thousands rather than the hundreds of individuals employed to date. Using 2,873 families, we were unable to confirm a recently published association of the interleukin 12B gene in 422 type I diabetic families. These results emphasize the need for large datasets, small P values and independent replication if results are to be reliable.     

The effect of reserpine on the Achilles reflex in normal young men

Zusammenfassung Reserpin in kleinen Dosen über eine Periode von 28 h führt zu einer Beschleunigung des Achillessehnenreflexes. Der Effekt ist statistisch gesichert, jedoch gering im Vergleich mit Veränderungen, die durch Schilddrüsenstörungen hervorgerufen werden.     

Diets of sympatric bobcats and coyotes during years of varying rainfall in central Arizona

We collected fecal samples (scats) of sympatric bobcats ( Lynx rufus ) and coyotes ( Canis latrans ) between 2000 and 2003 in a 53,600-ha area of the Upper Sonoran Desert in central Arizona. Our objective was to investigate composition, diversity, and overlap of diets of bobcats and coyotes in relation to varying rainfall in the Sonoran Desert of central Arizona. In general, bobcats ate more rodents than did coyotes, and coyotes ate more lagomorphs, large prey, and fruit/seeds than did bobcats. Composition of bobcat diets was independent of differences among years in annual rainfall and seasonal rainfall during summer–autumn (May–October) and winter–spring (November–April). Composition of coyote diets also was independent of drought conditions among years during summer–autumn, but coyotes ate more large prey and fewer rodents during years with winter–spring drought. Seasonally, bobcats ate more rodents than did coyotes in summer–autumn and winter–spring, whereas coyotes ate more lagomorphs than did bobcats during winter– spring, and more large prey and fruit/seeds in both seasons. Coyotes ate more large prey and lagomorphs during winter–spring, when seasonal rainfall was higher, and more fruit/seeds in summer–autumn, when seasonal rainfall was lower. Diversity of diets was consistently higher for coyotes than for bobcats, and increased for bobcats but not for coyotes during winter–spring drought and during higher seasonal rainfall in winter–spring. Overlap of diets between predators was independent of rainfall levels. We suggest that bobcats in the Sonoran Desert are more selective, specialized predators and that coyotes are more generalist, opportunistic predators. We hypothesize that, although diversity of bobcat food items and composition of coyote diets differ with varying rainfall in the Sonoran Desert, patterns of feeding strategy are independent of seasonal differences in precipitation and effects of drought, and bobcats and coyotes partition food resources independently of varying rainfall.     

Persistence of Mycobacterium tuberculosis in macrophages and mice requires the glyoxylate shunt enzyme isocitrate lyase

Mycobacterium tuberculosis claims more human lives each year than any other bacterial pathogen. Infection is maintained in spite of acquired immunity and resists eradication by antimicrobials. Despite an urgent need for new therapies targeting persistent bacteria, our knowledge of bacterial metabolism throughout the course of infection remains rudimentary. Here we report that persistence of M. tuberculosis in mice is facilitated by isocitrate lyase (ICL), an enzyme essential for the metabolism of fatty acids. Disruption of the icl gene attenuated bacterial persistence and virulence in immune-competent mice without affecting bacterial growth during the acute phase of infection. A link between the requirement for ICL and the immune status of the host was established by the restored virulence of delta icl bacteria in interferon-gamma knockout mice. This link was apparent at the level of the infected macrophage: Activation of infected macrophages increased expression of ICL, and the delta icl mutant was markedly attenuated for survival in activated but not resting macrophages. These data suggest that the metabolism of M. tuberculosis in vivo is profoundly influenced by the host response to infection, an observation with important implications for the treatment of chronic tuberculosis.