William Burke - US grants

Bacillus sphaericus strain 1593 has been proven effective as a microbial insecticide for control of larvae of several species of disease-vectoring mosquitoes. This bacterium kills mosquito larvae by means of a protein toxin associated with sporulating cells and mature spores. The usefulness of this organism as a biocontrol agent is enhanced by its abililty to recycle and persist in nature for long periods of time. Recently, a plasmid transformation system has been developed for this organism which will be used to manipulate the genes effecting insecticidal toxin production, toxicity, and host range. Techniques fundamental to recombinant DNA methodology will be adapted for use with this bacterium in order to accomplish these genetic modifications. The genetic information specifying the extremely potent larvicidal protein produced by a related bacterium, Basillus thuringiensis var. israelensis, will be cloned in Bacillus subtilis and ultimately transferred into B. sphaericus strain 1593.

The long term aims of this project are 1) to determine whether 3,4 dihydroxy-pheylglycolaldehyde (DOPEGAL) is an endogenous neurotoxin; 2) to determine whether in Alzheimer's disease (AD) the concentration of DOPEGAL increases in epinephrine (Epi) nerve cell bodies where it has access to DNA and where binding to DNA could decrease transcription of the mRNAs of functionally important proteins. Four experiments are proposed: 1. The concentration of DOPEGAL and enzymes involved in its synthesis will be measured in Epi neuronal cell bodies in AD and controls using gas chromatography-mass spectrometry and radiochemical assays. 2. The mRNA levels of catecholamine synthesizing enzymes will be measured in Epi neuronal cell bodies in AD and controls using polymerase chain reaction (PCR) and quantitative autoradiography. 3. In vivo toxicity will be determined by injecting DOPEGAL onto norepinephrine neurons in the rat locus ceruleus. Histologic evidence of toxicity will be determined by counting neurons and measuring cross sectional areas of neurons identified immunohistochemically. Chemical evidence of toxicity will be determined by measuring activity and amounts of catecholamine (CA) synthesizing enzymes using radiochemical and immunochemical assays, CA enzyme mRNA levels will be measured using polymerase chain reaction with autoradiography and quantitative densitometry. 4. The precise identity of the neurotoxin and whether its effects are mediated by extracellular cell surface receptor mechanisms or by purely intracellular mechanisms of toxicity will be determined in vitro using cultured adrenal medullary explants. Evidence of toxicity will be determined using the above histological and chemical procedures. In addition, HPLC-EC measurements of the MAO-A metabolites will be made to determine their uptake into cells. CA uptake blockers will be used to determine if effects are mediated by intracellular mechanisms. Neuronal death is the anatomic basis for clinical symptoms in AD yet cellular and subcellular mechanisms underlying neuronal degeneration are poorly understood. The Epi neurotransmitter system is affected late in AD. While Epi neurons are undergoing degeneration there is no cell loss making it an ideal system to look at the biochemical mechanisms of neuronal degeneration.

This project will give students hands-on experience with computer-interfaced instrumentation by incorporating fourier transform infrared (FT-IR) spectroscopy into six undergraduate chemistry courses. In General Chemistry, the instrument will be used to study isotopic substitution and to correlate the spectrum of an unknown compound with those in a computerized database. Applications in Organic Chemistry include structural interpretation and identification of reaction products, compounds isolated from natural sources, and real life samples (e.g., cinnamon and mouthwash). Polymers synthesized by students will be characterized by FT-IR. In Instrumental Analysis, students will record IR spectra of single strands of hair in a methods development experiment and will determine the percentage of ethanol in gasohol samples. Other FT-IR experiments include a kinetics study of a ligand substitution reaction in Physical Chemistry and enzyme-substrate interactions in aqueous solution in Biochemistry. The experience gained by students will give them skills to apply to other computer-interfaced instruments and valuable training for graduate school and the workforce.

9353997 Awad The two week seminar, "FIBER OPTICS FOR ENGINEERING TECHNOLOGY" is designed to introduce 30 faculty members from community and undergraduate colleges to the fundamental principles and applications of optical fiber technology, especially as related to engineering technology education at the two and four year level. The seminar/workshop is planned for June, 1994 with follow-up activities throughout the following academic year and beyond. The program is a collaborative effort involving a four year technical college and three community colleges in the Boston area. Eighty hours of training will be offered, half of which will be in the fiber optic laboratory, and the remainder consisting of lectures, simulation and analysis using computers as well as visits to local companies that use fiber optic technology. Substantial hands-on involvement by industry representatives is integral to the program. The laboratory exercises are designed to allow participants to handle optical fibers and several of the specialized instrumentation and test equipment developed specifically for fiber optics. Women, minorities and faculty who intend to introduce/enhance fiber optics at their home institutions will receive priority. No prior knowledge of fiber optics or communications is required. Follow-up outreach to regional community colleges is planned for the summer of 1995. ***

DESCRIPTION (provided by applicant): Treatment for head and neck cancer (HNC) is arduous and debilitating. As a result, up to 40% of patients develop depression which is rarely recognized or treated. Untreated depression in cancer patients lengthens hospital stays, and reduces adherence with medical treatment, quality of life, and survival. Patients with HNC typically develop depression within the first 3 months of their diagnosis. Developing depression can lead to interruptions or delays in cancer treatment which in turn may diminish the overall prognosis. Depression can also lead to suicide;patients with cancers of the larynx and tongue account for 2 of the 3 highest suicide rates in cancer patients. We are proposing a groundbreaking approach to the problem of depression in HNC. Patients with HNC have the greatest risk for depression in the several months after diagnosis and during treatment for their cancer. We seek to prevent major depression before it can develop by initiating prophylactic treatment with the antidepressant citalopram soon after the diagnosis of HNC is made, thus potentially avoiding many serious sequelae including treatment interruption or delays. Depression also has an enormous impact on quality of life in patients with HNC. Additionally, an overwhelming majority of HNC patients use alcohol and tobacco, and despite adverse effects on prognosis, it is often difficult for them to decrease use of these substances. We will therefore evaluate whether prophylactic use of citalopram can preserve quality of life during and after treatment for HNC and reduce alcohol and tobacco use by these patients. We are therefore proposing a randomized, double blind, placebo-controlled trial investigating the prophylactic utility of the antidepressant citalopram in nondepressed patients who are about to begin treatment for newly diagnosed or recurrent head and neck cancer. We will evaluate whether such prophylactic treatment can significantly reduce the development of major depression, improve adherence to cancer treatment, preserve quality of life and reduce tobacco and alcohol use. This proposal has considerable relevance to public health in that it not only tests whether depression can be prevented in patients with HNC during treatment but also examines whether this intervention can also improve timely completion of the cancer therapy. Additionally, we will examine whether the quality of life of such patients can be better preserved during treatment and whether alcohol and tobacco use can be diminished. If successful, this approach may be applicable to other medical illnesses that have high rates of comorbid depression.