Antiproliferative Effects of a Triterpene-Enriched Extract from Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), on Human Lung Cancer Cells.

Ganoderma lucidum, a mushroom that has been used to treat disease in East Asia for centuries, has been shown to be effective against many types of tumors, but the exact cellular mechanism of action is unknown. In this study we examined proliferation of a lung cancer cell line after treatment with 12 concentrations of powdered G. lucidum for 24, 48, and 120 hours. Based on half-maximal inhibitory concentrations values, proliferation of the H1793 cell line seemed to be sensitive to the extract in a time- and dose-dependent manner. We used immunoblot analysis to examine the amounts of cell cycle proteins (cyclin D, Cdk4, and Cdc2) and apoptotic proteins (Bcl-xL and Bax) after treatment with a range of G. lucidum concentrations. Changes in amounts of proteins that regulate the cell cycle were consistent with longer G1 and G2 phases. Proapoptotic protein (Bax) levels increased 6.5-fold, with a commensurate increase in the Bax-to-Bcl ratio, especially at 48 and 120 hours. These results suggest that the decrease in cellular proliferation correlated with a change in both cell cycle progression and apoptosis, and that the triterpenoid in G. lucidum is the bioactive component. Further biochemical characterization of this ancient herbal remedy could hold promise for treating lung cancer.

Transgenic apple expressing an antigenic protein of the human respiratory syncytial virus.

A gene coding for the human respiratory syncytial virus (RSV)-F protein, driven by the constitutively expressed CaMV 35S promoter, was introduced into leaf tissues of apple, Malusxdomestica Borkh. cv. Royal Gala, via Agrobacterium-mediated transformation. Two putative transgenic lines were identified, and the presence of the RSV-F gene was confirmed by polymerase chain reaction (PCR). A total of 25 plants from these different transgenic events were successfully rooted, acclimatized, and transferred to the greenhouse. Stable integration of the transgene was confirmed and transgene copy number was determined by DNA gel blot analysis. Expression of the npt-II selectable marker and RSV-F was determined using reverse-transcription polymerase chain reaction (RT-PCR). Furthermore, enzyme-linked immunosorbent assay (ELISA) revealed varying levels of protein expression of the RSV-F transgene, ranging from 0 to 20 microg/g tissue. This is a first step in an effort to assess the efficacy of using apple for developing a plant-based vaccine against RSV.

Effectiveness of a cloning and sequencing exercise on student learning with subsequent publication in the National Center for Biotechnology Information GenBank.

With rapid advances in biotechnology and molecular biology, instructors are challenged to not only provide undergraduate students with hands-on experiences in these disciplines but also to engage them in the "real-world" scientific process. Two common topics covered in biotechnology or molecular biology courses are gene-cloning and bioinformatics, but to provide students with a continuous laboratory-based research experience in these techniques is difficult. To meet these challenges, we have partnered with Bio-Rad Laboratories in the development of the "Cloning and Sequencing Explorer Series," which combines wet-lab experiences (e.g., DNA extraction, polymerase chain reaction, ligation, transformation, and restriction digestion) with bioinformatics analysis (e.g., evaluation of DNA sequence quality, sequence editing, Basic Local Alignment Search Tool searches, contig construction, intron identification, and six-frame translation) to produce a sequence publishable in the National Center for Biotechnology Information GenBank. This 6- to 8-wk project-based exercise focuses on a pivotal gene of glycolysis (glyceraldehyde-3-phosphate dehydrogenase), in which students isolate, sequence, and characterize the gene from a plant species or cultivar not yet published in GenBank. Student achievement was evaluated using pre-, mid-, and final-test assessments, as well as with a survey to assess student perceptions. Student confidence with basic laboratory techniques and knowledge of bioinformatics tools were significantly increased upon completion of this hands-on exercise.