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Related post: □ (a) HUMAN SUBJECTS n (al) MINORS □ (a2) INTERVIEWS n (b) HUMAN TISSUES (c) NEITHER SUMMARY OF WORK (200 words or less - underline keywords) Studies of aerobic energy metabolism in parasites and mammals were continued and devoted primarily to elucidating the sequence of electron transfer in the respiratory chains of Entamoeba histolytica and Giardia Iambi ia . Efforts were focused primarily on the iron-sulfur proteins in these parasites. The sub- cellular distribution and molecular properties of the Fe.S centers in E^. histolytica differ from those observed in G. Iambi ia. Reexamination of L- serine metabolism in E, histolytica was necessary when it was found that the amebae no longer oxidize this amino acid. Detailed studies of this paradox have shown that the current strain cultivated in Diamond ' s new TYI-S-33 medium fail to respond to L-serine, whereas the old strain frozen since 1975 and re- cultivated in the old TP-S-1 medium , readily oxidize the amino acid as reported previously. Studies on the ATPase of Leishmania donovani kinetoplasts disclosed that these organelles readily hydrolyze ATP in the presence of ^ ion . Mamma - lian studies were continued on the effect of heme depletion on the respiration of cultured murine erythroleukemia cells . Walker ascites tumor cells , and iso- lated rat h epato cytes. 25-29 PHS-6040 (Rev. 10-76) Serial No. ZOl AI 00098 _24 LPD Project Description : The object of this project is to conduct fundamental studies on the mechanisms of aerobic energy metabolism in mammalian and parasitic organisms. Currently these studies center primarily on identification and characterization of components of the respiratory chains of pathogenic, enteric protozoa, and elucidation of the pathway of electron flow to molecular oxygen. Other studies focus on bioenergetic phenomena in organel- les isolated from protozoa, mammalian tissues and cultured tumor cells. Tissues and cells are disrupted mechanically and subcellular fractions isolated by differential and gradient centrifugations. Cellular and sub- cellular constituents, enzyme activities, and metabolic pathways are determined, characterized and elucidated by enzymatic, chemical, radio- chemical and physical methods. Changes in oxygen concentration are determined polarographically with the Clark electrode. Heme depletion in rat hepatocytes is accomplished pharmacologically by Generic Finasteride perfusion of 4,6-dioxoheptanoic acid. Parasite studies : Continued investigation of the respiratory chains of Entamoeba histolytica and Giardia lamblia cultivated axenically in TYI-S-33 medium have revealed marked differences in the iron-sulfur electron transfer proteins Order Finasteride Online of these parasites. Upon cell fractionation, the bulk of respiratory enzymes and electron carriers are found in the cytosol of E, histolytica , and in the particulate fraction of G^. lamblia . Electron paramagnetic resonance (EPR) studies have shown the presence of multiple iron-sulfur centers in E. histolytica that exhibit characteristic "signals" after enzymatic reduction with NADPH. These centers appear to be of the f erredoxin-type that have low redox potentials. In contrast, the EPR signal obtained with G. lamblia becomes evident only in the oxidized state, and closely resembles the high potential Fe.S center of the Chromatium type. This difference in the properties of the Fe.S centers of the protozoa may be related to the large difference in Cheap Finasteride their habitats (Weinbach, Claggett, Keister and Kon) . In earlier work (1975) we first observed that L-serine markedly stimulated 0„ uptake of E. histolytica . Dr. Takeuchi, a visiting associate, extended this observation, and found that L-serine was converted to pyruvate, and that the 0_ uptake was due to oxidation of this keto acid. After these findings were published (1979) we found that L-serine no longer stimulated the respiration of E. histolytica . Detailed examination of Buy Finasteride Online this paradox is helping to resolve Finasteride Online the riddle. Our earlier work was done with trophozoites cultivated in Diamond's original TP-S-1 monophasic medium containing Panmede, an ox liver papain digest. Subsequently, Diamond, et^ al. formulated a new medium TYI-S-33 containing yeast extract and iron salts which is superior in supporting growth of the axenized amebae. Currently, all of our studies are done with amebae cultivated in the new medium. These parasites do not oxidize L-serine, but specimens, which had been kept frozen and recultivated in the old 25-30 Serial No. ZOl AI 00098-24 LPD medium, oxidized L-serine as we had originally observed. This is an interesting example of the effect of the cultivation medium on the biology of a parasite (Weinbach, Claggett, Keister and Diamond). Continued studies on the respiratory metabolism of Giardia Iambi ia have revealed the presence of multiple enzymes which catalyze the oxidation of reduced pyridine nucleotides Buy Finasteride (NAD(P)H) . These are found both in the cytosol and particulate fractions Order Finasteride of disrupted cells. The particulate Purchase Finasteride Online enzyme tranfers electrons from NAD(P)H to molecular oxygen without the need for exogenous carriers, indicating intactness of the respiratory chain. Attempts to sub-fractionate the particles by differential or density gradient centri- fugations showed Buy Cheap Finasteride that the enzymatic activity was not associated with discrete organelles. Oxidation of NAD(P)H in the soluble fraction was slow, but greatly increased upon addition of menadione or other electron acceptors. Studies with inhibitors of electron transport, disclosed dif- ferences in the properties of the soluble and particulate enzymes. The physiological significance of these results is that enzymes which oxidize reduced pyridine nucleotides are vital by providing a mechanism for the reoxidation of NADH generated during glycolysis by an organism that lacks both mitochondria and lactate dehydrogenase (Weinbach and Keister) . Studies on mitochondrial bioenergetics were resumed this Purchase Finasteride year by characterizing the ATPase of Leishmania donovani kinetoplasts. Similar to its mammalian counterpart, the enzyme is activated by Mg ion, and strongly inhibited by micromolar amounts of oligomycin. Paradoxically, the latent ATPase activity
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