Author: Eldon D. Nielson Biological Research Division To: Dr. Murray Senkus Director of Research Re: MONTHLY RESEARCH REPORT Biological Research Division 1966, No. 12 ec TN Yu Mey ly MRR-B, 1966, No. 12 January 4, 1967 Period Covered: November 21 through Deceuber 30, 1966 4 No. of Pages: 3 BIOLOGICAL TESTING Snoke Mildness Studies The results of an experiment comparing the mildness scores of WINSTON cigarettes with competitive brands, employing an equal tar weight basis, were reported last month. A statistical analysis of those results showed that Kent and L&M cigarettes scored significantly lover on the mildness test than the other cigarettes, consisting of WINSTON, Chesterfield Filter, Viceroy, Lucky Strike Filter, and Pall Mall Filter cigarettes. This study is being repeated to determine the scores when the cigarettes are smoked on the basis of equal numbers of cigarettes enoked to equal butt length G-7 cigarettes containing magnesium citrate and magnesium malate were compared with regular G-7 cigarettes for irritancy scores. The results again demonstrated that cigarettes containing the magnesium malate and citrate are milder than the control G-7 cigarettes. IL. Pharmacology of Mariolide-type Compounds Several compounds structurally related to mariolide have been examined for pharmacologic activity. Of the series, dihydromariolide is the most toxic and also the most potent analeptic agent. None of the other compounds tested demonstrate biological activity as interesting as that of dihydro- mariolide or mariolide in the tests employed. IIL. Smoking Machine Ten rats were exposed to smoke, employing the smoking machine built in our laboratories. It was observed that the machine leaked too much suoke to be satisfactory and that the exposure chamber should be reduced in size. Changes are being made on the machine to correct these deficiencies. : IV. Biological Testing of Compounds ‘The biological testing of RJR compounds and the biological evaluation of abscisin are continuing. No new noteworthy biological activities were observed in this period. 6EE9 vEz705Effect of Nicotine on Excretion of NcMethyInicotinamide Studies are continuing on the effect of smoking on tryptophan metabolism. A study was made to determine whether nicotine is the component in smoke responsible for the observed effects. Rats were treated with different levels of nicotine, and the urine was collected and analyzed for N-methyl- nicotinamide. Increasing doses of nicotine appeared to cause an increase in the excretion of N-methylnicotinamide, which would tend to confirm an effect of nicotine on the metabolism of tryptophan. Another possible explanation would be that some metabolite of nicotine interferes with the determination of N-methylnicotinamide. B._ STARCH BIOCHEMISTRY 1._Glucose Isomerase Studies have been undertaken to determine the optimum conditions for the growth of culture 2453 and the effect of these conditions on the production of glucose isomerase. The microorganisn grew moderately well in a medium containing 2% xylose and snall anounts of tryptone and yeast extract; the growth phase was completed in about 48 hours. In a 2% xylose plus corn steep solids medium, growth was continuing after 96 hours of incubation; and a proportionally higher cell yield was obtained. However, the isomerase activity per unit cell weight was lover in the rapidly growing cultures. Cells of culture 2453 have been used ten times in succession to convert glucose to fructose. The initial conversion produced about 40% fructose, and the tenth conversion produced about 26% fructose after a 24-hour incubation period. This is an interesting reflection on the stability of the enzyme preparation. The main rationale for a patent on the enzymatic conversion of glucose to fructose by R. 0, Marshall was that his enzyme preparation did not convert glucose to fructose at concentrations below 0.2 molar. In preliminary experiments, culture 2453 was found to be operative below this low concentration limit. It is believed that this constitutes another basis for believing that this process will not be covered by the Marshall patent. CIGARETTE BEETLE CONTROL Preliminary data have been obtained on the effect of five different treatments of tobacco storage sheds for the control of the tobacco beetle. A summary of the preliminary data show that all of the treatment procedures were better than no treatment at all. The most effective means of control. of the tobacco beetle appears to be a combination of sealing the shed and ona oAzostreating with a DDVP aerosol. It will be necessary to continue this study until the tobacco has completed its aging period in order to give a definitive evaluation of the treatments. Distribution: Dr. W. M. Bright Mr, Manford R. Haxton Dr. Murray Senkus Mr. Donald F. Mackintosh Dr. Richard E. Farrar Dr. Karol J. Mysels Mr. E. He Harwood Dr. Eldon D. Nielson Library (2) Dr. Charles W. Nystrom Dr. William C. Squires Submitted: January 4, 1967 Completed: January 4, 1967 From manuscript: Approved: M.S. JAN 6 i827 tne9 oszos