Illinois Junior Academy of Science - Yearbook (Urbana, IL)

Page 31 text:

-, after the injection of insulin, and thus no apparent genetic change. 27 There is a greater concentration of carbohydrates in the membranes around the neural tube, noto- chord, and yolk sac after insulin injection, and a decreased concentration in the tail bud region. 3l Rumplessness seems to be caused by the action of insulin in stopping the yolk sac from transferring glycogen to the embryo, or preventing metabolic processes, and this deficiency causes the undeveloped tail bud. 43 The naturally occurring mutation of rumplessness tCreeper conditionl is due to the genes' interfering with the carbohydrate metabolism and transport in the tail bud of the embryo. The Effects of Hyperbaric Oxygenation on Cerebral Metabolism STEVEN TRIPPEL In tests determining the ability of mice to successfully run through a maze, it has been found that when the run was made in a chamber of high pressure oxygen, the mice ran the maze both faster and with fewer errors than did mice under normal pressure air. It is suggested that this is a result of an increase in cerebral metabolism and a proced- ure for experimentation to test this has been developed. Hyperbaric oxygenation has been in use for clinical pur- poses for a long time and many of its physiological effects are thus, well known and understood. The possibilities of its having any mental effect, however, have not, to my know- ledge, ever been explored. Yet, in the use of HBO fhyper- baric oxygenl in the medical treatment of certain diseased aged people, there has been observed a substantial loss of senility during and shortly following the period of oxygena- tion? Although these observations could possibly be due to a generally improved physical condition resulting from the curing of the disease, it poses an interesting question whether or not they may be due, at least in part, to the HBO itself. In this report, then, my experiments testing this question are analyzed and a method is proposed for evaluating the tentative answer to it. To test whether or not HBO has any effect on mental activity, a number of mice were trained to run through a maze in order to obtain food. They were trained by being fed only in the maze and only after having successfully run through it. Motivation was supplied to them by feeding them only once a day and thus, by keeping them hungry. The trained mice were then divided into two groups: a con- trol group and an HBO groupf In testing, each mouse was placed at the entrance to the maze and allowed to proceed as it pleased. Its progress was timed and the number of errors twrong turnsl were recorded. In the case of the HBO group, this maze run was made inside a hyperbaric chamber at three atmospheres iabsolutel ygressure of pure oxygen. The control was run in the same maze but under normal atmospheric conditions. The results of running each group through the maze twice with approximately and hour and a half interval be- tween runs showed that the HBO group ran the maze in a shorter average time and with an average of fewer errors than did the control group. tSee graphs and tables for spe- cific resu1ts.l This seems to indicate that HBO makes mice smarter, or at least more alert than they are under normal oxygen partial pressure inormal pO,l. This one set of tests, however, is by no means sufficient to determine whether or not HBO actually does have the standard effect of increasing mental alertness. Even if such extensive tests were made Originally there were four groups: a control, a one- atmosphere pure oxygen group, a two-atmosphere pure HBO group, and a three-atmosphere pure HBO group. The four groups together consisted of forty-seven trained mice, and were to determine whether there was an increase in mental activity corresponding to the increase in pO,. Unfortunately, however, a dog kept in the same hospital lab with these mice escaped from its cage and devoured every one of them. The ten mice that could be obtained afterward were not suffi- cient to make four new groups. so as to show beyond reasonable doubt that such effects are the result of increased oxygen pressure, they still would give no indication of just why and how: thus, a more thorough examination of the present results is called for. The following graphs and tables illustrate the results of the above-described experimentation. un hu no ll:-ulsoxoa TIHE 'M Gm-.5 M u ' - ll' Rn.. Aung. ' on-.ll neva,-,- XP I I i l Lac' R... A-a-UV, . ' .QI X kiss in-an-a :Hia Is Yiwu. llvnhu- Kia Qvn ,, A TuMElcoNraoLp:'::-fl-M. use H ' TQ 'Qf','2'f, M .... .. at l J N i i W H U rd' R-nt Hvu-mme. A a c- n ' : ' Hun-. Latin ani lun T E S 'I' co urlox. 2 mer suewcs lsr RVN Frau. 6134... :vt Ru- ao --. 3 7... we Ov'--dl as 50 zz The mice under three atmospheres tabsolutel pure oxygen iunder fifteen times the normal oxygen partial pressure! ran the maze an average of about twenty-two percent faster than the control mice under normal atmospheric conditions. w ERRORSBTESTHf.'T5I'w'mi Ell0l.S 3 A 1 . 1 R-an Average 1'4 lun marqu- Q 1 1. l -.t l k-- r 1. l ? ' I 'Z ' 3 xW 35' Hausa Nuubu- mul Run Q Emmons 5 coNTnoLlLi.'::.l-fa Q. YKRORS 2. L A ln Q Annan I I '-VA R-la huermge, O I a. 1 at rs a x 'z Q' A B C D E Nusa Leiter and Run I Run -'l I-6 44, ar' R... -'ff - 9 al TEST CONTROL. ZDIFFERENCE Ovev-all .93 l-2 31

Page 30 text:

Wave forms Minimum overshoot 30W 15W 0111 Maximum overshoot 27592 2052 0f1, Output voltage unloaded 60V unloaded 59.4V unloaded 60V vs. load .03W . . 42V .03W . . 59.4V 30W . . . 60V 2W .... 56V Efficiency about 15W about 3565 85011 or at .03W at .03W better about 7O'11 at 1.5W sidered in light with the increase in catalase activity, there is a possibility of the enzyme glucose oxidase being active in the tissue. This enzyme oxidizes glucose according to the diagram shown below. In normal tissue, gluconate forma- tion is only the first few steps in glucose metabolism. It is then further oxidized through the Kreb's Cycle to carbon dioxide and water. In aerobic dehydrogenation, the oxida- tion of glucose stops at the gluconate level. Flavin-adenine- dinucleotide QFADJ is the co-enzyme and functions as a hydrogen acceptor. It transfers two hydrogen atoms to a molecule of oxygen which forms hydrogen peroxide. The catalase then decomposes the peroxide as above. This, of course, is just a thedry concerning a possible Although the original objectives were not fully .achieved as of yet, much improved performance has been obtained. This .power supply is to be used for a 15W portable audio amplifier. A Possible Metabolic Alteration in Neoplast Plant Tissue JOHN LAWLER Spalding Institute Peoria Sponsor - Rev. Daniel Reardon, C. S. V. Crown gall is a malignant tumor which affects plants. There are several different causitive agents, such as insects, viruses, nematodes, bacteria, and perhaps others. The tumors I work with are induced in the common sunflower CHelian- thus annuusl by the bacterium Agrobacterium tumefaciens, strain B6. The purpose of the work is to determine if there is any alteration in the metabolic process of the tumor. The part of the Work described in my paper is the initial research on this problem. The first part of the research was to assay catalase activity in the tumors. Catalase is present in most living organisms. Its function is to break down hydrogen peroxide to oxygen and water, because of its toxicity, accoring the equation: 2H,O2 I 2H2O + O, A manometer based on the Warburg Manometer was built to measure changes in gas pressure and volume. For the catalase activity, a substrate of .O6'11 hydrogen peroxide in phosphate buffer, pH 7.0, was used. Young Helianthus seedlings were inoculated with the bacteria and the tumors were allowed to develop to the desired size. A control of the same size and age, only just wounded and not inoculated, was used. When the tissue was to be tested, .1 gram of the tissue was cut and sliced into thin sections and placed in 2 ml. of the substrate. Measurements of the increase of oxygen were made every minute for five minutes and then graphed. The results tan example shown in graph Al showed nearly a twofold increase in catalase activity by the crown gall than by the control plant. The next step is described below. This step is to determine any correlation between cata- lase activity and glucose oxidation. To do this, the mano- meter is again used. The substrate this time is four parts of a 1111 glucose solution added to eleven parts of a Ringer- phosphate buffer in a ratio of 1O:1. The manometer system is filled with oxygen and the tissue to be tested. .1 gram, is added to 2 ml. of the Ringer-phosphate-glucose. The decrease in oxygen level is recorded and graphed every minute for five minutes. The results fa sample given in graph Bl show a marked decrease in oxygen consumption by as much as 50611 in crown gall tumors. In considering this information, we must recall that the tumor tissue is much more active in cell division than normal tissue, but it is seen here that the tumor does not use as much oxygen as the normal tissue. In considering this, Tamaoki et al. 119601 thought this could be a result of Al more efficient use of energy by the tumor cell or BJ a variance of the tissue in its metabolic pathways. When con- change in the enzyme and metabolic activities of crown gall. I I have now started work involving colorimetric study of glucose oxidation and other metabolic intermediates. AEROBIC DEHYDROGENATION H-1 - C - O H . A O H1 F-OH H fi- X OH EXT O xcfon cl '2H HN! HX' OHIXI NH CQH I-ICSO ' c-of f X C - Cf 1 I OH I l H HO H OH glucose gluconolactone H C OH H2 C-OH 2 o C'H l - Hx! l ,OH H H xx C OH H C I O X fC OH H C I O !0H f OHN I lf C -- C E3-'E OH I I H OH H OH gluconolctone gluconic acid il 2H + FAD I FADH2 FADH2 -I- O2 I H,O, + FAD 2H,O. I 2H.O -l- O1 The Specificity of Insulin-Induced Rumplessness in Chicken Embryos THOMAS ZAZECKIS St. Mel High School Chicago Sponsor - Mr. Thomas Gorell The investigation's purpose was to determine the speci- ficity of insulin in causing the phenocopy of rumplessness in White Leghorn embryos and to relate this information to naturally occurring mutations to determine their method of operation. Altogether 234 White Leghorn eggs were used, 138 were injected with .05 ml. Illetin Q2 units insulinl and 96 were injected with .05 ml. sterile distilled water as a control. The embryos were split into 2 groups, the first incubated for 48 hours and the second for 72 hours. Both sets were then prepared for microscopic study ffixed, microtomed, stained! using both Periodic Acid-Schiff's Reagent CPASJ and Pyronin-Methyl Green IPMGJ for the staining process. The slides were then studied and the following conclusions ar- rived at: ll There is no noticeable change in the nucleic acids

Page 32 text:

The mice under three atmospheres Cabsolutel 100 percent oxygen ffifteen times the pO, of the controll ran the maze with an average of about 30 percent fewer errors than did the control mice. These results seem to indicate that mice under three atmospheres partial pressure of oxygen are able to function both faster and more accurately-than those under the normal one-fifth atmosphere oxygen partial pressure. In analyzing these results, however, it must'be recognized that they are not statistically as valuable as they should be if any con- clusions are to be drawn from them. This is a result of the fact that each mouse was run only twice and that there was a shortage of mice due to the loss of the original set. Thus these results must not be regarded as showing that HBO does definitely increase mental activity, but rather as hinting that this might be the case. They are, however, ade- quate to warrant further experimentation that could better test the possible merit of the idea that HBO increases mental activity. In addition, it should be noted that under HBO, there exist certain physiological conditions pertinent to this ex- periment which are independent of the results obtained but which seem to support them. For example, one of the recog- nized effects of subjecting mammals to HBO is an increased oxygen content of the blood. This, in accordance with Hen- ry's law lmoles of gas dissolved I k x gas pressurel, is because the blood plasma dissolves oxygen in quantities pro- portionate to its partial pressure lpO,J. Thus, in this experi- ment, the plasma oxygenation was increased fifteen fold and the overall blood oxygenation by about 31 percent. lThe slight overall increase relative to the large plasma increase is a result of the fact that the blood's hemoglobin, whose amount of oxygen is not much affected by pressure increases, carries the large majority of the blood's oxygenfl. It is interesting to note that the overall difference in blood oxygenation C31 percentl between the test and control groups in these experiments is very close to the overall dif- ference in efficiency of their maze runs f27fZ:J. The significance of this increased pO of the blood is that it means that when an animal such as a mouse for a humanl is put under HBO, its tissues, including the brain tissues, are bathed in the extra oxygen from the blood plasma. This suggests that the ixidative processes consti- tuting the metabolism of such tissues as the brain could increase under HBO conditions. The experimentation done thus far serves as an empirical observation which might support the hypothesis that HBO increases brain metabolism in that it shows how mental activity could be affected by HBO. However, since, as mentioned previously, this experi- ment's results are statistically somewhat cloudy, and because it does not test the hypothesis directly tthis present hypo- thesis was formulated after these results were obtainedl, further experimentation is required. For this reason, I have developed the following procedure and am making arrange- ments for its execution. After a good deal of formulating and collecting of ideas, I decided that the best feasible means of testing whether HBO increased cerebral metabolism is to measure the arterio-venous difference in the quantity of oxygen in the blood of the major artery and vein serving the brain. Briefly, this amounts to performing an operation on the neck of an animal such as a dog and attaching to each its internal carotid artery and internal jugular vein the electrode of a gas analyzer' and the probe of a flowmeterf From the re- sults obtained by these two instruments, the A-V difference in oxygen quantity can be determined. The gas analyzer is an instrument designed to measure the pressure of such gases as oxygen and carbon dioxide in the tissues of animals. It operates by means of an electrode sheathed in a thin gas-permeable membrane which is inserted directly into the tissue fblood in this easel to be analyzed. The presence of gases at the electrode tip initiates a current which is amplified and calibrated on a meter and which gives the partial pressure of the gas at the electrode tip. The electrode can be inserted into the artery Cveinl through the use of a T tube as shown. 2 rm, T .TuT ' mini! ini -- M -.N Hr-fu-y fveinl august Ti' The flowmeter is an instrument designed to measure the rate of flow fvolume per unit time! of a liquid through -L a tube such as an artery or vein. It does this by measuring the voltage produced when flowing blood fwhich is slightly ionizedl cuts the electromagnetic field produced by a probe encompassing the artery fveinl. This voltage is proportion- ate to the rate at which the field is cut and thus also to the rate of blood flows. The probe can be placed on the artery fveinl as shown. 62 The absolute quantity iexpressible in molesl of oxygen pass- ing a given point per unit time in the artery and the vein can be determined from the product of the flowmeter read- ings lin volumeftimel and the gas analyzer readings fin at- mospheres of oxygen at a given temperaturel through a series of relatively simple calculations. The difference in moles of oxygen found between the in- ternal cartoid artery and internal jugular vien will give a good approximation of the amount of oxygen being consumed by the brain i.e. brain metabolism. lIt will be but an approxi- mation because there are other minor arteries and veins feeding and emptying the brain whose services cannot be measured. The i. carotid and i. jugular would have to be the only vessels to the brain and would have to serve only the brain if the results were to be a precise measurement of cerebral metabolism.l The hypothesized results, then, should show the amount of oxygen consumed by the brain under EEO conditions to be greater than that under normal con- 'tions. The experiments testing this will be run with a control group consisting of dogs anesthetized and operated upon fwith instruments attachedl but under normal atmospheric conditions, and with a test group of dogs anesthetized, operated upon, and under HBO. In an effort to reduce the variables introduced by such factors as anesthetic effect, etc., the two groups will consist of as similar dogs as can possibly be obtained. In addition, each group will serve as both a control and a test against itself in that after readings on the control group have been made, it will be tested under HBO to see how the readings change. Similarly, after readings have been made with the test group under HBO, it will be made to serve as control. This not only means that each group will serve as a control both against itself and the other group, but also that any effect of time on the physical condition of the operated dogs will be averaged out by the alternation of oxygenation. It was not until just recently that I gained access to the equipment necessary to carry out the above-described procedure, apd as a result, the tests necessary to confirm the hypothesis that HBO increases cerebral metabolism have not yet been made. However, tests run to determine the possibility of HBO having an effect on mental activity in- dicate a degree of likelihood that this further experimenta- tion will confirm this hypothesis. Footnotes and Acknowledgements 1. From the work of Dr. J. Williams of Sunninghill, England. 2. See page two of this paper. 3. Boerema, I., Meijne, N. G., Brummelkamp, W. K. Bouma, S., Mensch, M. H., Lamarmans, F., Hanf, M. S. and Van Aaleren, W.: Life without blood. J. Thoracic and Cardiovascular Surgery. 1: 133, 1960. 4. My thanks to Dr. Paul Nora of Northwestern University Medical School, Chicago, Illinois, for permitting me to use his gas analyzer and to Mr. Frank Cieslak for demonstrating its operation. 5. My thanks to Dr. F. John Lewis of Northwestern Uni- versity Medical School, Chicago, for making available to me his flowmeter and for showing me how to use it. 6. Cordell, R. A. and Spencer, M. P.: Electromagnetic Blood Flow Measurement in Extracorporeal Cir- cuits. Annals of Surgery Vol. 151 No. 1. pp. 71-74. January 1960.