Multiple human papillomavirus types replicate in 3A trophoblasts.

Human papillomavirus (HPV) are more prevalent in spontaneous abortions than elect abortions and preferentially infect the trophoblasts. Related to this, HPV type 16 has been shown to productively replicate in 3A trophoblasts in tissue culture. Extending these earlier studies, the described study addresses the issue whether other genital HPV types (11, 18, and 31) can replicate in trophoblasts. In determining this, HPV-11, 18, or 31 genomic DNAs were lipofected into 3A trophoblasts in culture, thus finding all three HPV types could de novo DNA replicate in 3A trophoblasts (Southern blot) and sequentially express their early and late genes as RNA (RT-PCR) and as protein (immunohistochemistry for L1). HPV-transfected 3A lysates from all three HPV types were also shown to contain HPV infectious units by infection of normal skin raft cultures and by neutralization by specific antibody. Furthermore, microarray analysis revealed the gene expression profile of normal keratinocytes (NK) was closer to 3A trophoblasts than to normal fibroblasts. Moreover, the critical HPV transcription factors AP-1 and Sp1 were found to be more highly expressed in 3A cells than NK. These findings suggest trophoblasts, like squamous epithelium, are broadly permissive for HPV, and some similarities in the gene expression repertoire of these two cell types are consistent with this. Finally, these data support our previous results that demonstrate the relationship between HPV infection of the trophoblast and spontaneous abortions.

Bioinformatics and proteomics approaches for aging research.

Aging is a natural phenomenon that affects the entire physiology of an organism. Elucidating the molecular mechanisms underlying this complex process remains a major challenge today. Humans make poor models for research into aging because of their long life span. Thus, most of the current knowledge is through studies conducted in lower organisms. Large differences in life spans make it difficult to extrapolate the results of experiments carried out in model organisms to humans. Recent advances in genomic and proteomic technologies now permit generation of data pertaining to aging on a large-scale. In addition, several web-based community resources and databases are available that provide easy access to the available data. Use of bioinformatics and systems biology type of approaches provide a framework to start dissecting this complex biological phenomenon. Here, we discuss various genomic, transcriptomic and proteomic approaches that have the potential to provide a comprehensive mechanistic insight into the aging process.

Human protein reference database as a discovery resource for proteomics.

The rapid pace at which genomic and proteomic data is being generated necessitates the development of tools and resources for managing data that allow integration of information from disparate sources. The Human Protein Reference Database (http://www.hprd.org) is a web-based resource based on open source technologies for protein information about several aspects of human proteins including protein-protein interactions, post-translational modifications, enzyme-substrate relationships and disease associations. This information was derived manually by a critical reading of the published literature by expert biologists and through bioinformatics analyses of the protein sequence. This database will assist in biomedical discoveries by serving as a resource of genomic and proteomic information and providing an integrated view of sequence, structure, function and protein networks in health and disease.

Ultrastructural studies on nodules induced by pyrimidine auxotrophs of Sinorhizobium meliloti.

Twenty three pyrimidine auxotrophs of Sinorhizobium meliloti Rmd201 were generated by random mutagenesis with transposon Tn5. On the basis of biochemical characters these auxotrophic mutants were classified into car, pyrC and pyrE/pyrF categories. All auxotrophs induced white nodules which were ineffective in nitrogen fixation. Light and electron microscopic studies revealed that the nodules induced by pyrC mutants were more developed than the nodules of car mutants. Similarly the nodules induced by pyrE/pyrF mutants had more advanced structural features than the nodules of pyrC mutants. The nodule development in case of pyrE/pyrF mutants was not to the extent observed in the parental strain. These results indicated that some of the intermediates and/or enzymes of pyrimidine biosynthetic pathway of S. meliloti play a key role in bacteroidal transformation and nodule development.

Isolation and symbiotic characterization of aromatic amino acid auxotrophs of Sinorhizobium meliloti.

Ten aromatic amino acid auxotrophs of Sinorhizobium meliloti (previously called Rhizobium meliloti) Rmd201 were generated by random mutagenesis with transposon Tn5 and their symbiotic properties were studied. Normal symbiotic activity, as indicated by morphological features, was observed in the tryptophan synthase mutants and the lone tyrosine mutant. The trpE and aro mutants fixed trace amounts of nitrogen whereas the phe mutant was completely ineffective in nitrogen fixation. Histology of the nodules induced by trpE and aro mutants exhibited striking similarities. Each of these nodules contained an extended infection zone and a poorly developed nitrogen fixation zone. Transmission electron microscopic studies revealed that the bacteroids in the extended infection zone of these nodules did not show maturation tendency. A leaky mutant, which has a mutation in trpC, trpD, or trpF gene, was partially effective in nitrogen fixation. The histology of the nodules induced by this strain was like that of the nodules induced by the parental strain but the inoculated plants were stunted. These studies demonstrated the involvement of anthranilic acid and at least one more intermediate of tryptophan biosynthetic pathway in bacteroidal maturation and nitrogen fixation in S. meliloti. The alfalfa plant host seems to provide tryptophan and tyrosine but not phenylalanine to bacteroids in nodules.