How are neuronal thermosensitivity and lack of thermoreception related in the duck's hypothalamus? A tentative answer

1. 1.|In 15 conscious Pekin ducks, 40 “warm sensitive” hypothalamic neurons were identified according to their discharge rates at 40°C T_hy (F₄₀), local temperature coefficients (Δ/ΔT) and Q₁₀.

2. 2.|Q₁₀ and either F₄₀ or ΔF/ΔT were little or not related.

3. 3.|A positive correlation between F₄₀ and ΔF/ΔT was observed which was particularly close (r = 0.94 and 0.96) when the neurons were classified according to their Q₁₀ of <2 and >2.

4. 4.|The results suggest that neurons with positive temperature coefficients in the duck's hypothalamus mostly exhibit linear to exponential temperature-discharge relationships.

5. 5.|This is an contrast to observations on mammalian hypothalamic thermosensitive neurons and may relate to the absence of the thermosensory function in the duck's rostral brainstem.

Limiting steps in Photosystem II and water decomposition in Chlorella and spinach chloroplasts

1. 1. By varying the redox potential of a chloroplast suspension, we obtained new evidence for an equilibrium between states S₀ and S₁ in the model of Kok, B., Forbush, B. and McGloin, N. (1970, Photochem. Photobiol. 11, 457–475). The mid-point potential of the S₀ to S₁ couple is close to that for the pool of the electron acceptor of System II, A to A⁻.

2. 2. The limiting steps between two consecutive photoreactions of System II in Chlorella and spinach chloroplasts, have been studied.

2.1. (a) The limiting step from S₁ to S₂ (noted γ₁ (Δt)) is not exponential. Its temperature coefficient becomes greater as the reaction proceeds. The shape of the kinetics is an intrinsic property of each center. Chloroplasts fixed with 2% glutaraldehyde, show simple first order kinetics.

2.2. (b) The limiting step from S₀ to S₁ (γ₀ (Δt)) exhibits the same characteristics as γ₁ (Δt)).

2.3. (c) The limiting step from S₂ to S₃ (γ₂ (Δt)) shows sigmoidal kinetics; two reactions are involved. One of the reactions exhibits the same properties as γ₀ (Δt) and γ₁ (Δt).

2.4. (d) The limiting step from S₃ to S₀ (γ₃ (Δt)) is a first order reaction, two times slower than the other transitions. This reaction is interpretated in terms of oxygen release.

3. 3. We also studied the limiting steps in the presence of low concentrations (50 μM) of hydroxylamine. The results favor the binding of two molecules of hydroxylamine to every photochemical center.

Day-night variation of thermoregulatory responses to intraperitoneal electric heating in rats

1. 1.Hypothalamic temperature (T_hy), nonevaporative heat loss (R + C + K), evaporative heat loss (E), thermal conductance (k), metabolic heat production (M) and heat storage (S) of rats were simultaneously measured by direct and indirect calorimetry during internal heat loading (2 W per rat) with an intraperitoneal electric heater.

2. 2.The tests were made twice a day; once during the day (1000–1200 h) and once at night (2200–2400 h) at an ambient temperature of 24°C.

3. 3.The resting values of T_hy, colonic temperature, (R + C + K), E, M and heart rate, and the T_hy threshold for tail skin vasodilation (T_th) during internal heat load were significantly higher at night than during the day.

4. 4.The slopes showing the relationshiop between (R + C + K), k or M and T_hy were significantly steeper during the day than at night after T_hy exceeded T_th.

5. 5.The slopes showing the relationship between E or S and T_hy were not different during the day and at night.

6. 6.These results indicate that the responses of nonevaporative heat loss and heat production to internal heat load vary with the time of day in rats.

Mechanism of respiration-driven proton translocation in the inner mitochondrial membrane. Analysis of proton translocation associated with oxidation of endogenous ubiquinol

A study is presented of the kinetics and stoichiometry of fast proton translocation associated to aerobic oxidation of components of the mitochondrial respiratory chain.

1. 1. Aerobic oxidation of ubiquinol and b cytochromes is accompanied in EDTA particles, obtained by sonication of beef-heart mitochondria, by synchronous proton uptake.

2. 2. The rapid proton uptake associated to oxidation of ubiquinol and b cytochromes is greatly stimulated by valinomycin plus K⁺, but is unaffected by carbonyl cyanide p-trifluoromethoxyphenylhydrazone.

3. 3. 4 gion H⁺ are taken up per mol ubiquinol oxidized by oxygen. This H⁺/2e− ratio, measured in the rapid anaerobic-aerobic transition of the particles is unaffected by carbonyl cyanide p-trifluoromethoxyphenylhydrazone.

4. 4. In intact mitochondria aerobic oxidation of oxygen-terminal electron carriers is accompanied by antimycin-insensitive synchronous proton release, oxidation of ubiquinol and reduction of b cytochromes. The amount of protons released is in excess with respect to the amount of ubiquinol oxidized.

5. 5. It is concluded that electron flow along complex III, from ubiquinol to cytochrome c, is directly coupled to vectorial proton translocation. The present data suggest that there exist(s) between ubiquinol and cytochrome c one (or two) respiratory carrier(s), whose oxido-reduction is directly linked to effective transmembrane proton translocation.

The b-type cytochromes of bovine heart mitochondria: Absorption spectra, enzymatic properties, and distribution in the electron transfer complexes

1. 1. Three b-type cytochromes (b_557.5, b₅₆₀, and b_562.5), plus a chromophore with an absorption peak at 558 nm at 77 °K, have been found to be associated with the electron transport system of bovine heart mitochondria. The reduced minus oxidized spectra of these components at 77 °K, as well as that of cytochrome c₁, have been recorded with a wavelength accuracy of ± 0.1 nm and presented to the nearest 0.5 nm. All the major and β absorption peaks of cytochromes b_557.5, b₅₆₀, b_562.5, c₁ and c have been shown by fourth derivative analysis to be present in the dithionite-reduced minus oxidized spectra of mitochondria and submitochondrial particles.

2. 2. The distribution of the above components has been studied in the four electron transfer complexes of the respiratory chain. Cytochromes b₅₆₀, b_562.5 and c₁, as well as chromophore-558, were found to fractionate into Complex III (reduced ubiquinone-cytochrome c reductase), whereas cytochrome b_557.5 was found in Complex II (succinate-ubiquinone reductase).

3. 3. Cytochrome b₅₆₀ was readily reduced by NADH or succinate, but b_562.5 was not reduced by substrates unless the preparation was treated with antimycin A. In antimycin-treated preparations pre-reduction of c₁ with ascorbate inhibited the subsequent reduction of b_562.5 by substrates. These results indicate that b₅₆₀ and b_562.5 correspond, respectively, to b_K and b_T previously described by Chance et al.¹⁴ (1970, Proc. Natl. Acad. Sci. U.S. 66, 1175–1182).

4. 4. Similar to b₅₆₀, chromophore-558 can be reduced by substrates in the absence or presence of antimycin A. However, in antimycin-treated preparations, pre-reduction of c₁ inhibits its subsequent reduction by substrates. This property is similar to that of b_562.5.

5. 5. Cytochrome b_557.5, which occurs in Complex II, appears to have a low mid-point potential. It can be reduced with dithionite and oxidized by fumarate or ubiquinone. CO treatment of dithionite-reduced b_557.5 neither modified the spectrum of this cytochrome nor diminished the extent of b_557.5 reoxidation by fumarate.

6. 6. Antimycin A treatment does not appear to alter the spectra of the above cytochromes. However, small amounts (< 4%) of ethanol or methanol, which are usually added to particles as solvent for antimycin A, have a pronounced effect on the peaks of cytochrome c₁. The spectrum of cytochrome c₁ at 77 °K as modified by 3% (v/v) ethanol is shown.

The influence of temperature on the behaviour of the Mexican jumping bean

1. 1.|The jumping behaviour of the Mexican jumping bean (Laspeyresia solitans) is stereotyped and predictable and has the characteristics of behaviour which is under the control of a central programme.

2. 2.|Temperatures above and below about 25°C decrease the length of time that movements occur. The movements stop relatively abruptly and not gradually. The duration of movements is apparently controlled by an internal timer whose duration of operation is influenced by temperature.

3. 3.|The frequency of movements show instantaneous temperature compensation from about 21 to 45°C (Q₁₀ = 1.23), whereas between 15 and 21°C there is a large temperature dependence (Q₁₀ = 21.9).

The phosphorylation potential generated by respiring mitochondria

1. 1. The phosphorylation potential, ΔG_P = ΔG₀′ + 1.36 log ([ATP]/[ADP][P_i]), where ΔG_O′ is the standard free energy of hydrolysis of ATP at a given pH, and [ATP], [ADP] and [P_i] refer to concentrations in the suspending medium, has been determined in rat-liver mitochondria under various conditions.

2. 2. The ATP/ADP ratio is relatively constant, over a 10-fold range of phosphate concentration. Thus, the phosphate potential is higher at low phosphate concentration. State-4 rat-liver mitochondria in the presence of succinate, oxygen and low concentrations of phosphate in State 4 maintain a phosphorylation potential of 16.1 kcal (67.3 kJ) per mole ATP.

3. 3. High concentrations of ATP inhibit ADP uptake, and it is suggested that this is the reason for the independence of the ATP/ADP ratio on the phosphate concentration. A steady-state ratio is set up dependent upon two processes that are relatively slow compared with State-3 respiration, namely ADP transport and ATP hydrolysis.

4. 4. The phosphorylation potential calculated from the concentrations of total ADP, ATP and P_i within State-4 mitochondria is 4.5 kcal/mole less than that in the suspending medium.

5. 5. It was shown experimentally that the phosphorylation potential cannot be calculated from the ΔG of the redox couple, the respiratory-control ratio and the P:O ratio, as has been suggested in the literature.

6. 6. The measured phosphorylation potential is 83% of that calculated from the span succinate to oxygen, assuming thermodynamic equilibrium, and 95% of that calculated from the span NADH to oxygen.

7. 7. Based on the measurements of the phosphorylation potential and of the redox potentials and redox states of redox components in mitochondria, ubiquinone and cytochrome b are found at their expected position at the junction of the phosphorylations at Sites 1 and 2. The iron-sulphur centres 2 and 5 and the iron-sulphur centre of succinate dehydrogenase also probably lie at this junction. Cytochrome a₃ lies at its expected junction between phosphorylation Sites 2 and 3. A number of electron carriers (cytochromes c, c₁, and a, the iron-sulphur centre of Complex III and the EPR-detectable copper), however, lie in the ‘no-man's land’ within Site 2.

8. 8. A phosphorylation potential of 16.1 kcal/mole corresponds to a membrane potential of 350 mV in State 4, on the basis of the chemiosmotic hypothesis.

Binding of radioactively labeled carboxyatractyloside, atractyloside and bongkrekic acid to the ADP translocator of potato mitochondria

1. 1. The inhibition of the ADP-stimulated respiration of potato mitochondria by carboxyatractyloside is relieved by high concentration of ADP or by the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP). Atractyloside is a much less potent inhibitor than carboxyatractyloside. The inhibition of the ADP-stimulated respiration required about 60-times more atractyloside than carboxyatractyloside.

2. 2. [³⁵S]carboxyatractyloside and [³H]bongkrekic acid bind to potato mitochondria with high affinity (K_d = 10 to 20 nM, n = 0.6–0.7 nmol per mg protein). Added ADP competes with carboxyatractyloside for binding; on the contrary ADP increases the amount of bound bongkrekic acid. [³H]atractyloside binds to potato mitochondria with a much lower affinity (K_d = 0.45 μM) than carboxyatractyloside or bongkrekic acid.

3. 3. Bound [³H]atractyloside is displaced by ADP, carboxyatractyloside and bongkrekic acid. The displacement of bound [³⁵S]carboxyatractyloside by bongkrekic acid and of bound [³H]bongkrekic acid by carboxyatractyloside is markedly increased by ADP.

4. 4. Bongkrekic acid competes with [³⁵S]carboxyatractyloside for binding. Addition of a small concentration of ADP considerably enhances the inhibitory effect of bongkrekic acid on [³⁵S]carboxyactratyloside binding.

5. 5. The adenine nucleotide content of potato mitochondria is of the order of 1 nmol per mg protein. ADP transport in potato mitochondria is inhibited by atractyloside 30- to 40-times less efficiently than by carboxyatractyloside.

Acetylene reduction with physiological electron donors by extracts and particulate fractions from nitrogen-fixing Azotobacter chroococcum

1. 1. Preparations were obtained from Azotobacter chroococcum which reduced acetylene to ethylene using physiological electron donors instead of sodium dithionite. These preparations fell into two categories: those which required catalytic amounts of benzyl viologen for acetylene reduction and those that did not.

2. 2. Acetylene reduction without benzyl viologen or sodium dithionite was observed only with particles that sedimented at 40 000 × g after disrupting bacteria in the French press or with preparations obtained by disrupting bacteria protected by a mixture of defatted bovine serum albumin-Ficoll-MgCl₂ with liquid N₂; supernatant fractions required benzyl viologen for acetylene reduction.

3. 3. Added ATP inhibited acetylene reduction by large particles; ATP and MgCl₂ were necessary for maximum acetylene reduction with bovine serum albumin-protected preparations.

4. 4. NADH and carbon substrates acted as electron donors but H₂ did not; NAD⁺ was necessary for maximum acetylene reduction with carbon substrates.

5. 5. Anaerobic conditions were necessary for maximum acetylene reduction in all cases.

The P-700-chlorophyl a-protein complex and two major light-harvesting complexes of Acrocarpia paniculata and other brown seaweeds

Acrocarpia paniculata thylakoids were fragmented with Triton X-100 and the pigment-protein complexes so released were isolated by sucrose density gradient centrifugation. Three main chlorophyll-carotenoid-protein complexes with distinct pigment compositions were isolated.

1. (1) A P-700-chlorophyll a-protein complex, with a ratio of 1 P-700: 38 chlorophyll a: 4 ta-carotene molecules, had similar absorption and fluorescence characteristics to the chlorophyll-protein complex 1 isolated with Triton X-100 from higher plants, green algae and Ecklonia radiata.

2. (2) An orange-brown complex had a chlorophyll a : c₂ : fucoxanthin molar ratio of 2 : 1 : 2. This complex had no chlorophyll c₁ and contained most of the fucoxanthin present in the chloroplasts. This pigment complex is postulated to be the main light-harvesting complex of brown seaweeds.

3. (3) A green complex had a chlorophyll a : c₁ : c₂ : violaxanthin molar ratio of 8 : 1 : 1 : 1. This also is a light-harvesting complex.

The absorption and fluorescence spectral characteristics and other physical properties were consistent with the pigments of these three major complexes being bound to protein. Differential extraction of brown algal thylakoids with Triton X-100 showed that a chlorophyll c₂-fucoxanthin-protein complex was a minor pigment complex of these thylakoids.     

Descending influences from nucleus raphe magnus on fusimotor neurone activity in rats

1. 1.Single fusimotor fibres were isolated in the ventral roots of lumbosacral segments of urethane-anaesthetized rats, and effects of electrical stimulation of the nucleus raphe magnus (NRM) on their spontaneous activity were investigated. The experiments were carried out in rats whose bilateral preoptic and anterior hypothalamic regions (PO/AH) were electrolytically destroyed to eliminate the influences of these regions to fusimotor activity.

2. 2.Of 44 fusimotor fibres studied, 38 (86%) were found to be affected by NRM stimulation. The effects of NRM stimulation were classified according to their response pattern: primary depression (D-type, n = 24), facilitation followed by depression (F-D-type, n = 5) and primary facilitation (F-type, n = 8). The most predominant effect of NRM stimulation upon fusimotor activity was characterized by a strong depression followed by a complete cessation of firing lasting either for a short period or for more than 30 min (D- and F-D-type).

3. 3.In three fusimotor fibres studied in the different preparations, it was observed that a NRM-evoked depression response was blocked by an intraperitoneal administration of a serotonin antagonist, p-chlorophenylalamine (p-CPA) (10 mg/kg).

4. 4.The results indicate that the NRM exerts descending inhibitory or facilitatory influences on fusimotor neurones, and suggest that cold shivering is controlled by modulating fusimotor neurone activity via the serotonergic raphe-signal pathways.

Expression of NADPH-diaphorase in nucleus tractus solitarius after peripheral injection of E. coli endotoxin in rats

1. The possibility that peripheral exogenous pyrogens can activate brainstem nuclei by abdominal viscera afferents was studied using the NADPH-diaphorase method in E. coli lipopolysaccharide treated rats with and without ibuprofen pre-treatment.

2. NADPH-d staining revealed: (i) a significant increase in the number of NADPH-d labeled neurons in the subpostremal area of the nucleus tractus solitarius lipopolysaccharide treated rats compared with control animals (p<0.05), and (ii) an almost similar number of NADPH-d positive neurons in both control and ibuprofen pre-treated rats (p=0.091).

3. These data indicate that peripheral administration of an exogenous pyrogen stimulates the synthesis of nitric oxide in the nucleus tractus solitarius, and are consistent with the hypothesis of a direct neural link between the periphery and the hypothalamus.

ATPase complex and oxidative phosphorylation in chloramphenicol-induced megamitochondria from mouse liver

1. 1. Functional properties of the ATPase complex are investigated in megamitochondria isolated from livers of weanling mice fed a diet containing 2% chloramphenicol, as an inhibitor of mitochondrial protein synthesis.

2. 2. Whereas the specific activity of ATPase remains unchanged in chloramphenicol-induced megamitochondria, about 40% of the enzyme activity is resistant to inhibition by oligomycin, triethyltin or venturicidin. It is concluded that the ATPase complex lacks one or more components whose synthesis or accumulation is dependent on mitochondrial translation. The inhibitor-resistant ATPase portion appears tightly bound to the mitochondrial membrane.

3. 3. Respiratory chain phosphorylation is tightly coupled in isolated megamitochondria. ATP synthesis and ATP-P_i exchange are diminished by 40%, as compared to control mitochondria, but both processes are sensitive to oligomycin, triethyltin or venturicidin.

4. 4. The decrease in ATP synthesis and ATP-P_i exchange in megamitochondria correlates quite well with the emergence of inhibitor-resistant ATPase.

5. 5. The following electron transport activities in the megamitochondria are reduced: NADH-cytochrome c reductase, by 60%, cytochrome oxidase, by 80%; the amount of antimycin required to gain complete inhibition of the bc₁-segment is diminished by more than 50%. On the other hand succinate dehydrogenase activity is increased by 50%.

6. 6. Chloramphenicol-induced megamitochondria appear to be a useful system for studying the role of mitochondrial translation in the assembly of mammalian mitochondria.

Thermal conductivity and H2O content in rabbit kidney cortex and medulla

1. 1.|H₂O content and local-tissue thermal conductivity were measured in cortex and medulla of 7 freshly-excised rabbit kidneys.

2. 2.|Tissue H₂O content and thermal conductivity k (83.4% and 0.516 W m^−t K⁻¹, respectively) in the medulla were significantly higher than those (77.7% and 0.475 W m⁻¹ K⁻¹, respectively) measured in the cortex.

3. 3.|Correlations between the measured parameters are made, and the variability of previously-reported measurements of kidney-tissue thermal conductivity is discussed.

The effect of 4-methyl-histamine on temperature regulation in the rat

1. 1. The histamine H₂-antagonist, 4-methyl-histamine causes a fall in body temperature when injected into the third ventricle of the rat.

2. 2. The hypothermia is prevented by prior intraventricular injection of an H₂-antagonist, metiamide, but not by an H₁-antagonist, pyrilamine.

3. 3. The fall in temperature is accompanied by increased cutaneous heat loss.

4. 4. These data support the concept that there are H₂-receptors in the central thermoregulatory pathways of the rat.

Algorithm for application of Fourier analysis for biorhythmic baselines of pharmacodynamic indirect response models

The change of an indirect pharmacological response R(t) can be described by a periodic time-dependent production rate k_in (t) and a first-order loss constant k_out. If k_in(t) follows some biological rhythm (e.g., circadian), then the response R(t) also displays a periodic behavior. A new approach for describing the input function in indirect response models with biorhythmic baselines of physiologic substances is introduced. The present approach uses the baseline (placebo) response R_b(t) to recover the equation for k_in(t). Fourier analysis provides an approximate equation for R_b(t) that consists of terms (usually two or three) of the Fourier series (harmonics) that contribute most to the overall sum. The model differential equation is solved backward for k_in(t), yielding the equation involving R_b(t). A computer program was developed to perform the square L²-norm approximation technique. Fourier analysis was also performed based on nonlinear regression. Cortisol suppression after inhalation of fluticasone propionate (FP) was modeled based on the inhibition of the secretion rate k_in(t) using ADAPT II. The pharmacodynamic parameters k_out and IC₅₀ were estimated from the model equation with k_in(t) derived by the new approach. The proposed method of describing the input function needs no assumption about the behavior of k_in(t), is as efficient as methods used previously, and is more flexible in describing the baseline data than the nonlinear regression method. (Chronobiology International, 17(1), 77-93, 2000)     

Variations in the activity of fatty ACYL-CoA desaturases and electron-transport components in Tetrahymena microsomes with temperature and age of culture

1. 1.|Alterations in the fatty acid composition of microsomes were most marked in the exponential phase of both 39.5- or 15°C- grown Tetrahymena pyriformis NT-1.

2. 2.|Activities of palmitoyl-CoA and stearoyl-CoA desaturases were lower in 15°C cells than in 39.5°C cells, while the activity of oleoyl-CoA desaturase was higher in 15°C cells.

3. 3.|Activities of the terminal component of the desaturation system as well as all three desaturases (palmitoyl-CoA, stearoyl-CoA, oleoyl-CoA) were higher in the exponential phase than in the stationary phase for cells grown at both temperatures.

4. 4.|NAD(P)H-cytochrome c reductase activity and cytochrome b₅ content were reduced whereas NADH-ferricyanide reductase activity was increased in the stationary phase at both 39.5 and 15°C.

Skin temperatures, thermosensory and pleasantness estimates in case of heterogeneity in the thermal environment

1. 1. Seven thermal conditions were imposed on male sitting subjects (slightly clothed: 0.6 clo).

2. 2. A thermal mannikin was also used to determine the exact operative temperature, T₀.

3. 3. Conditions were: uniform (UN: all parameters at 24.5°C, air velocity at 0.15 ms⁻¹), heated ceiling (HC at 45°C), heated floor (HF at 34°C), cold floor (CF at 14°C), two conditions of one cold wall at 6°C (CW1 and CW2 respectively with and without air temperature compensation) and increased air velocity (AV at 0.4 ms⁻¹).

4. 4. Local skin temperatures and answers to questionnaires were obtained.

5. 5. Skin temperature variations were affected by conditions and slight T₀ changes.

6. 6. Comfort judgments were fairly well related to T₀, especially when expressed as differences between actual non-uniform environment and the uniform one.

7. 7. It is concluded that, in case of non-uniform environments close to thermoneutral zone, thermal comfort or discomfort reflects the climate alterations better than the thermal sensation does.

Biochemical and biophysical studies on cytochrome aa3 VIII. Effect of cyanide on the catalytic activity

1. 1. Cyanide inhibits the catalytic activity of cytochrome aa₃ in both polarographic and spectrophotometric assay systems with an apparent velocity constant of 4·10³ M⁻¹·s⁻¹ and a K_i that varies from 0.1 to 1.0 μM at 22 °C, pH 7·3.

2. 2. When cyanide is added to the ascorbate-cytochrome c-cytochromeaa₃−O₂ system a biphasic reduction of cytochrome c occurs corresponding to an initial K_i of 0.8 μM and a final K_i of about 0.1 μM for the cytochrome aa₃−cyanide reaction.

3. 3. The inhibited species (a²+a₃³⁺HCN) is formed when a²⁺a₃³⁺ reacts with HCN, when a²⁺a₃²⁺HCN reacts with oxygen, or when a³⁺a₃³⁺HCN (cyano-cytochrome aa₃) is reduced. Cyanide dissociates from a²⁺a₃³⁺HCN at a rate of 2·10⁻³ s⁻¹ at 22 °C, pH 7.3.

4. 4. The results are interpreted in terms of a scheme in which one mole of cyanide binds more tightly and more rapidly to a²⁺a₃³⁺ than to a³⁺a₃³⁺.

Electron transport particles released upon lysis of spheroplasts of Escherichia coli B by Brij 58

The non-ionic detergent, Brij 58, has been shown to specifically lyse the cytoplasmic membrane of Escherichia coli. This communication examines the electron transport system in membrane fractions prepared from such lysates and compares this system to those prepared by mechanical procedures.

1. 1. The particulate fraction contained all of the respiratory carriers demonstrable in whole cells in essentially the same ratios although enriched by 3–5-fold over their concentration in whole cells.

2. 2. Succinate, formate, and NADH but not glutamate, malate, or dihydroorotate were actively oxidized by the particulate fraction.

3. 3. With the exception of the formate oxidase system which appeared to utilize a system bypassing flavoprotein and cytochrome b₁, the other enzymatic activities appeared to function primarily through normal electron transport routes.

4. 4. The NADH oxidase and succinoxidase systems were sensitive to inhibition by 2-n-heptyl-4-hydroxyquinoline-N-oxide and antimycin A. The results suggest that these inhibitors function at the level of cytochrome b₁.

5. 5. All three activities were sensitive to complete inhibition by CN⁻.

6. 6. The results obtained from inhibitor studies coupled with the results obtained from examination of steady state, anaerobic, and chemical reduction of the respiratory pigments permitted a scheme for electron flow to be proposed.