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Biomedical subjects

Michael D Kane

Publications and source records attributed to Michael D Kane.

6 recordsLinked to original sources

A molecular beacon DNA microarray system for rapid detection of E. coli O157:H7 that eliminates the risk of a false negative signal.

A DNA hybridization based optical detection platform for the detection of foodborne pathogens has been developed with virtually zero probability of the false negative signal. This portable, low-cost and real-time assaying detection platform utilizes the color changing molecular beacon as a probe for the optical detection of the target sequence. The computer-controlled detection platform exploits the target hybridization induced change of fluorescence color due to the Förster (fluorescence) resonance energy transfer (FRET) between a pair of spectrally shifted fluorophores conjugated to the opposite ends of a beacon (oligonucleotide probe). Unlike the traditional fluorophore-quencher beacon design, the presence of two fluorescence molecules allows to actively visualize both hybridized and unhybridized states of the beacon. This eliminates false negative signal detection characteristic for the fluorophore-quencher beacon where bleaching of the fluorophore or washout of a beacon is indistinguishable from the absence of the target DNA sequence. In perspective, the two-color design allows also to quantify the concentration of the target DNA in a sample down to < =1 ng/microl. The new design is suitable for simultaneous reliable detection of hundreds of DNA target sequences in one test run using a series of beacons immobilized on a single substrate in a spatial format.

DNA, Bacterial↗

An information technology emphasis in biomedical informatics education.

Unprecedented growth in the interdisciplinary domain of biomedical informatics reflects the recent advancements in genomic sequence availability, high-content biotechnology screening systems, as well as the expectations of computational biology to command a leading role in drug discovery and disease characterization. These forces have moved much of life sciences research almost completely into the computational domain. Importantly, educational training in biomedical informatics has been limited to students enrolled in the life sciences curricula, yet much of the skills needed to succeed in biomedical informatics involve or augment training in information technology curricula. This manuscript describes the methods and rationale for training students enrolled in information technology curricula in the field of biomedical informatics, which augments the existing information technology curriculum and provides training on specific subjects in Biomedical Informatics not emphasized in bioinformatics courses offered in life science programs, and does not require prerequisite courses in the life sciences.

Biomedical Engineering↗

Induction of the transferrin receptor gene by benzo[a]pyrene in breast cancer MCF-7 cells: potential as a biomarker of PAH exposure.

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental DNA-damaging agents regarded as risk factors for human disease, including lung and breast cancer. The biotransformation of PAHs to carcinogenic metabolites is mediated by the aromatic hydrocarbon receptor (AhR), which activates transcription at xenobiotic responsive elements (XREs = 5'-GCGTG-3') found in the promoter regions of genes encoding for detoxifying enzymes, including CYP1A1 and CYP1B1. In this study, we wished to identify novel biomarkers that may be useful in monitoring critical carcinogenic events of the breast induced by PAHs. Using a GeneMAP CancerArray, we analyzed in breast cancer MCF-7 cells the temporal effects of the AhR agonist benzo[a]pyrene (B[a]P), which is a prototype PAH and known environmental carcinogen. Genes upregulated at least threefold by B[a]P and containing potential XREs within their promoter regions included CYP1A1, CYP1B1, paired box gene 3 (PAX3), cortactin (CTTN/EMS1), beta-2-microglobulin (B2M), and transferrin receptor (TfR). The stimulatory effects of B[a]P on expression of these genes were abrogated by cotreatment with the AhR antagonist flavonoid, alpha-napthoflavone (ANF). The TfR gene was selected for further analysis as its promoter region contains two potential XREs and its expression has been shown to be increased in breast cancer cells. Accumulation of TfR mRNA in B[a]P-treated cells was confirmed by quantitative real time PCR. Transient transfection studies indicated that the transcriptional activity of the TfR promoter was stimulated by B[a]P, whereas ANF counteracted this induction. These results indicate that the TfR gene may be a potential biomarker of PAH exposure.

Benzo(a)pyrene↗

Technical issues in DNA microarray production and utilization: impact on clinical research.

The adoption and utilization of genomic technologies in healthcare requires that many issues surrounding the integrity of these assay platforms, as well as their overall impact on human health, be definitive. Many concerns common to DNA microarray production must be addressed to exploit the high-content screening capabilities of this assay platform for both genotyping and gene expression profiling in the clinic. Equally important to the success of genomic technology in healthcare is the development of a supporting information system that offers impact at both the personal and population level, and facilitates the adoption, use and, ultimately, impact of genomic information in therapeutic decision support.

Base Sequence↗

3rd International Symposium on Early Toxicity Screening. 11 December 2002, Philadelphia, Pennsylvania, USA.

The 3rd International Symposium on Early Toxicity Screening: Early ADME-Tox Screening Approaches included presentations by primarily non-academic scientists, including biotechnology companies, pharmaceutical scientists and contract research service leaders. A central theme heard throughout the presentations involved the fundamental objectives of and obstacles to ADME and toxicity testing during drug development. In summary, successful preclinical drug safety assessment involves understanding a model's limited ability to mimic the human drug response, increasing the model's ability to mimic the human drug response, and adopting new strategies that model better human drug responses, while simultaneously conducting safety assessment more quickly, more thoroughly and more cost effectively.

Drug Evaluation, Preclinical↗

Aligning experimental design with bioinformatics analysis to meet discovery research objectives.

The utility of genomic technology and bioinformatic analytical support to provide new and needed insight into the molecular basis of disease, development, and diversity continues to grow as more research model systems and populations are investigated. Yet deriving results that meet a specific set of research objectives requires aligning or coordinating the design of the experiment, the laboratory techniques, and the data analysis. The following paragraphs describe several important interdependent factors that need to be considered to generate high quality data from the microarray platform. These factors include aligning oligonucleotide probe design with the sample labeling strategy if oligonucleotide probes are employed, recognizing that compromises are inherent in different sample procurement methods, normalizing 2-color microarray raw data, and distinguishing the difference between gene clustering and sample clustering. These factors do not represent an exhaustive list of technical variables in microarray-based research, but this list highlights those variables that span both experimental execution and data analysis.

Computational Biology↗