Influence of sucrose on the rheology and granule size of cross-linked waxy maize starch dispersions heated at two temperatures

Cross-linked waxy maize (CWM) starch dispersions (STDs) of concentration 50 g kg⁻¹ were heated in sucrose solutions containing 0–600 g kg⁻¹ (g sucrose/kg dispersion) at 85 °C at low shear and in intermittently agitated cans at 110 °C. The STDs heated in 0–300 g kg⁻¹ sucrose exhibited antithixotropic behavior, while those heated in 400–600 g kg⁻¹ sucrose exhibited thixotropic behavior. The mean starch granule diameter of the starch dispersions did not show strong dependence on sucrose concentration. The dispersions, especially those with high sucrose concentrations and heated at 110 °C, exhibited G′ versus frequency (ω) profiles of gels. The STDs exhibited first normal stress differences that increased in magnitude with the concentration of sucrose. Values of the first normal stress coefficient of canned dispersions calculated from dynamic rheological data plotted against ω and experimental values plotted against shear rate of some of the STDs overlapped.     

A method for in-vivo kinematic analysis of the forearm

PURPOSE: To develop a method for in-vivo kinematic study of normal forearm rotation using computed tomographic (CT) images and a custom apparatus which allows for control of amount of forearm rotation. METHODS: The forearm of one asymptomatic volunteer was CT-scanned in five positions: neutral, 60 degrees pronation, maximal pronation, 60 degrees supination, and maximal supination. Surface registration of the pronated/supinated image datasets with the neutral position was performed. The resulting transformation matrices were decomposed into finite helical axis (FHA) parameters. Kinematics were expressed as motion of the radius relative to the ulna. RESULTS: The axes of the forearm passed through the volar region of the radial head at the proximal radioulnar joint (PRUJ), extending towards the dorsal region of the ulnar head at the distal radioulnar joint (DRUJ). Distinct FHAs were calculated for each forearm position analyzed relative to neutral rotation. Forearm pronation FHAs were different from forearm supination FHAs. CONCLUSIONS: Our experimental methodology is capable of describing the in-vivo kinematics of the forearm with good accuracy and reliability. Future in-vivo studies would need to be performed using a larger sample size to further validate our preliminary results. An ideal clinical application of this methodology would be in the comparative study of patients with forearm dysfunction.     

瓶梗颈部螺旋且寄生卷叶蛾幼虫的被毛孢新种

Hirsutella tortricicola sp. nov.（菌株GZUIFR-hir090704）分离于寄生卷叶蛾科昆虫幼虫的真菌标本GZUIFR-kks-2。其主要形态特征为产孢细胞颈部具1-2个螺旋状结构且再育明显;分生孢子具黏液,橘瓣状,2.7-3.6×1.4-1.8μm。通过形态学比较和系统发育学分析确定H. tortricicola为被毛孢属新成员。     

X-ray absorption fine structure study of the atomic and electronic structure of molybdenum disulfide intercalation compounds with transition metals

The local structures of ‘host’ and ‘guest’ layers of MoS₂ intercalated with M(OH)₂ (M=Mn, Co and Ni) prepared via interaction of single-layer MoS₂ dispersions and solutions of M²⁺ salts were studied by X-ray absorption spectroscopy. According to M K-edge extended X-ray absorption fine structure (EXAFS) and X-ray absorption near-edge structure (XANES) results, the electronic structure and atomic environment of the M atoms in the intercalates are similar to that of the crystalline hydroxides M(OH)₂. In the Ni intercalate, Mo K-edge EXAFS revealed a structural change of the ‘host’ MoS₂ layers similar to that reported for water dispersions of MoS₂ single layers. S K-edge XANES data indicate that the change is associated with increased electron density on the S atoms in the matrix. SO₄²⁻ and Mo″⁻ (4 < n < 6) were detected in the intercalated materials exposed to air, suggesting that transition metal intercalation may increase the susceptibility of the MoS₂ layers to oxidation.     

Cofacial and antarafacial indenyl bimetallic isomers: a descriptive MO picture and implications for the indenyl effect on ligand substitution reactions

The hetero-bimetallic Ind complexes (Ind=indenyl anion) with the formula (CO)₃Cr(μ-Ind)RhL₂ (L=CO, ethylene; L₂=COD, COT, NBD) have two possible conformers, one with the metals at antipodal sides of the condensed bicyclic polyene (antarafacial, anti) and the other with both metals on the same side (cofacial, syn). These systems can be considered to contain 34 valence electrons by assuming that Ind is capable of donating all of its ten π electrons to the metals. A qualitative MO analysis, based on EHMO calculations, is presented. The anti conformer is compared to the homometallic 34e species, namely [CpRu(μ-Ind)RuCp]⁺. Here, the pseudo-symmetrical relationship between the two identical metal fragments strongly suggests that one Ind σ-bonding MO donates part of its electron density to each of the two metals, which thus become formally saturated (36e). This argument is extended to the anti Cr---Rh derivative in which a critical role is played by an empty FMO of the (CO)₂Rh(I) fragment with π symmetry (i.e. lying in the molecular mirror plane). The latter, in spit the large π_CO^* contributions, has good acceptor capabilities at the metal.The calculations suggest that the so-called indenyl effect, namely the increased rate of substitution of one CO ligand by another σ donor (e.g. a phosphine), although a structurally dynamic process, also depends on the electron density which accumulates on the aforementioned π FMO. In the series (η⁵-C₅H₅)Rh(CO)₂, (η⁵-C₇H₉Rh(CO)₂ and {η⁵-[(CO)₃CrC₇H₉]}-Rh(CO)₂ the progressively reduced donor capabilities of the cyclic polyene toward the (CO)₂Rh(I) fragment induce a larger electrophilicity in rhodium, in good agreement with the experimental data on substitution reactivity. Finally, the electronic features of the cofacial conformer (CO)₃Cr(μ-Ind)Rh(CO)₂ are examined. In spite of the long Cr---Rh separation (> 3 Å) the graphically illustrated MO analysis indicates a direct, heterodox, intermetallic linkage, which helps the metals to achieve a formal electronic saturation. In this case, the limited propensity of the complex toward any CO substitution reactions is likely attributable to the hindered mobility of the (CO)₂Rh(I) fragment.     

The effects of in vivo and in vitro non-enzymatic glycosylation and glycoxidation on physico-chemical properties of haemoglobin in control and diabetic patients

The erythrocyte deformability, which is related to erythrocyte internal viscosity, was suggested to depend upon the physico-chemical properties of haemoglobin. In the present study we employed ESR spectroscopy in order to explore further the extent to which the in vivo or in vitro glycation and/or glycoxidation might affect haemoglobin structure and conformation. We revealed that under both in vivo and in vitro conditions the attachment of glucose induced a mobilization of thiol groups in the selected domains of haemoglobin molecules (the increased h₊₁/h₀ parameter of maleimide spin label, MSL; 0.377 ± 0.021 in diabetics vs 0.338 ± 0.017 in controls, n = 12, P < 0.0001). The relative rotational correlation time (τ_c) of two spin labels, TEMPONE and TEMPAMINE, respectively, in erythrocyte insides (5.22 ± 0.42 in diabetics, n = 21 vs 4.79 ± 0.38, n = 16 in controls, P < 0.005) and in the solutions of in vitro glycated haemoglobin, were increased. Neither oxidation nor crosslinking of thiol groups was evidenced in glycated and/or oxidized haemoglobin. In addition, erythrocyte deformability was found to be reduced in type 2 diabetic patients (6.71 ± 1.08, n = 28 vs 7.31 ± 0.96, n = 21, P < 0.015). In conclusion, these observations suggest that: the attachment of glucose to haemoglobin might have decreased the mobility of the Lys-adjacent Cys residues, thus leading to the increased h₊₁/h₀ parameter of MSL. Such structural changes in haemoglobin owing to non-enzymatic glycosylation may contribute to the increased viscosity of haemoglobin solutions (r = 0.497, P < 0.0035) and the enhanced internal viscosity of diabetic erythrocytes (r = 0.503, P < 0.003). We argue that such changes in haemoglobin, and consequently in red blood cells, might contribute to the handicapped oxygen release under tissue hypoxia in the diabetic state.     

Oxygenation of Hexadecane in the Biosynthesis of Cyclic Glycolipids in Torulopsis Apicola

Biotransformation of [1-¹³C] labelled hexadecane, hexadecanol and hexadecanoic acid have been investigated using the yeast Torulopsis apicola. The yeast produces a microcrystalline mixture of two glycolipids, the lipophilic moiety of which consists of ω- or (ω-l)-hydroxylated hexadecanoic acid. Biosynthesis of these glycolipids takes place via hydroxylation of hexadecane, oxidation to hexadecanoic acid and ω or (ω-l)-hydroxylation of hexadecanoic acid. Feeding the cell cultures with a mixture of hexadecane and [1-¹³C] labelled hexadecane derivatives one observes ¹³C enrichment ratios which indicate that neither of the biohydroxylation or oxidation steps are rate limiting in the formation of the glycolipids, furthermore, two different monooxygenase systems appear to be involved in hydroxylation of hexadecane and hexadecanoic acid.     

The effect of two teeth resection procedures on the welfare of piglets in farrowing crates. Part 1

Teeth resection is a method of controlling the injurious effects of the aggression displayed when newborn piglets fight to establish a teat order. Recent European legislation discourages the practice. The objective of this study was to assess the effect of clipping and grinding piglets’ needle teeth, compared to leaving them intact, on the welfare of piglets in farrowing crates.

Six days pre-partum, 60 sows were assigned to one of three treatments. Litters had their teeth clipped (C), ground (G) or left intact (I) at birth. The time taken to carry out each procedure was recorded. Piglet weights and facial lesions, which were scored according to severity, were recorded on days 1, 4, 11, 18 and 27. Piglet weights were also recorded at birth. Mouth lesions were recorded on days 1, 4 and 27. Instantaneous scan samples of piglet behaviour were carried out for 30 min post-teeth resection procedure (1 min intervals), and for 6 h on days 1, 4, 8, 14, 21 and 26 (5 min intervals). One male and one female piglet per litter were chosen as focal animals and observed for 5 min each post-procedure and for 10 min each twice per day on days 1, 5, 12, 20 and 26. Mortalities were recorded throughout lactation.

Grinding took significantly longer than clipping the teeth or leaving them intact (F = 638.87, P < 0.001). I piglets had higher facial lesion scores than C and G piglets (F = 10.58, P < 0.001). A smaller proportion of piglets in I litters than C and G litters and a smaller proportion of piglets in G litters than C litters had at least one mouth lesion (F = 4.74, P < 0.001). During 30 min post-procedure, I piglets were active on the heatpad in more observations than C and G piglets (F = 3.49, P < 0.05). During 5 min post-procedure C piglets spent longer chomping than I piglets (F = 5.92, P = 0.05). On day 21, I piglets were active in more observations than G piglets (F = 2.11, P < 0.05). On day 26, G piglets were inactive in more observations than C and I piglets (F = 5.02, P < 0.05). On days 14 and 26, C piglets were sleeping in more observations than G piglets (F = 2.87, P = 0.05). There was a tendency for a larger proportion of I than C piglets to die due to overlying (F = 2.68, P = 0.08).

In conclusion, although all three options were associated with welfare problems, grinding can be recommended in preference to clipping or leaving the teeth intact.     

Stress relaxation function of bone and bone collagen

Relaxation Young's and shear moduli of bovine bone and bone collagen were investigated. It was found that each relaxation process observed had two stages, which were referred to as process I and process II in order of time. Process II was described by a simple exponential decay while process I was not. The Kohlrausch-Williams-Watts (KWW) function, ψ(t) = exp[t/τ₁)^B] (0 < B < 1), was found to be suitable to describe process I. The normalized relaxation modulus, M_r(t), was expressed by the combination of the simple exponential type relaxation function and the KWW function

M_r=A₁exp[−(t/τ₁)^B]+A₂exp[(t/τ₁)](0<B1)

On the basis of this equation, the relaxation mechanism in bone and bone collagen was identified. According to the model proposed for the KWW relaxation function, the stress relaxation process in bone was considered to be governed by viscoelastic properties of matrix collagen fiber. The model for the KWW relaxation function requires the disordered glassy structure of collagen fiber, which is consistent with the results of the structural investigations.     

Effects of peptide YY and its analogues on chloride ion secretion in fed and fasted rat jejunum

The aim of our study was to determine whether a meal modifies the antisecretory response induced by PYY and the structural requirements to elicit antisecretory effects of analogue PYY(22–36) for potential antidiarrhea therapy. The variations in short-circuit current (Isc) due to the modification of ionic transport across the rat intestine were assessed in vitro, using Ussing chambers. In fasted rats, PYY induced a dose- and time-dependent reduction in Isc, with a sensitivity threshold at 5 × 10⁻¹¹ M (ΔIsc −2 ± 0.5 μA/cm²). The reduction was maximal at 10⁻⁷ M (Isc −23 ± 2 μA/cm²), and the concentration producing half-maximal inhibition was 10⁻⁹ M. At 10⁻⁷ M, reduction of Isc by PYY reached 90% of response to 5 × 10⁻⁵ M bumetanide. The PYY effect was partly reversed by 10⁻⁵ M forskolin (Isc +13.43 ± 2.91 μA/h·cm², p < 0.05) or 10⁻³ M dibutyryl adenosine 3′,5′ cyclic monophosphate (Isc +12 ± 1.69 μA/cm², p < 0.05). Naloxone and tetrodotoxin did not alter the effect of PYY. In addition, PYY and its analogue P915 reduced net chloride ion secretion to 2.85 and 2.29 μEq/cm² (p < 0.05), respectively. The antisecretory effect of PYY was accompanied by dose- and time-dependent desensitization when jejunum was prestimulated by a lower dose of peptide. The antisecretory potencies exhibited by PYY analogues required both a C-terminal fragment (22–36) and an aromatic amino acid residue (Trp or Phe) at position 27. At 10⁻⁷ M the biological activity of PYY was lower in fed than fasted rats (p < 0.001). Our results confirm the antisecretory effect of PYY, but show that the fed period is accompanied by desensitization, similar to the transient desensitization observed in the fasted period with cumulative doses. This suggests that PYY may act as a physiological mediator that reduces intestinal secretion.     

Determination of bacterial cell number and cell volume by means of flow cytometry, transmission electron microscopy, and epifluorescence microscopy

Comparative measurements of bacterial total counts and volumes of flow cytometry (FCM), transmission electron (TEM), and epifluorescence microscopy (EFM), were undertaken during a four week mesocosm experiment. Total counts of bacteria measured by TEM, EFM, and FCM were in the range of 1 · 10⁶−6 cells ml⁻¹, 1 · 10⁶−3 · 10¹⁶ cells ml⁻¹, and 5 · 10⁵ cells ml⁻¹ respectively. The mean volume of the bacterial community, measured by means of EFM and TEM, increased from 0.12–0.15 μm³ at the start of the experiment to 0.39–0.53 μm³ at the end. Generally, there was good agreement between the two methods and regression analyses gave r = 0.87 (p < < 0.01) for cell volume and r = 0.97 (p < < 0.01) for cell number. DAPI stained bacteria with volumes less than 0.2 μm³ were not detected by flow cytometry and these were generally an order of magnitude lower than counts made by TEM and EFM. For samples where the mean bacterial cell volume was longer than 0.3 μm³, all three methods were in agreement both with respect to counts and volume estimates.     

Folded conformation of an immunostimulating tetrapeptide rigin: high temperature molecular dynamics simulation study

Employing high temperature quenched molecular dynamics (QMD) simulations the conformational energy space of an immunostimulating tetrapeptide rigin: H-Gly³⁴¹-Gln-Pro-Arg³⁴⁴-OH, is explored. Using distance dependent dielectric (=r_ij) 31 different low energy starting structures with identical sequence were computed for their conformational preferences. According to the hypothesis of O'Connors et al. [J. Med. Chem. 35 (1992), 2870], 83 low-energy conformers resulted from unrestrained molecular dynamics (MD) simulations, could be classified into two energy minimized families: A and B, comprised of 64 (Pro C^γ-endo orientation) and 19 (Pro C^γ-exo orientation) structures, respectively. An examination of these families revealed the existence of a remarkably similar folded backbone conformation: torsion angles being φ_i+1 ≈−65°, ψ_i+1 ≈−65°, φ_i+2 ≈−65°, ψ_i+2 ≈−60°, characterizing a distorted type III β-turn structure across the central Gln-Pro segment. The folded conformation of rigin is devoid of a classical 1 ← 4 intra-molecular hydrogen bond nevertheless, the conformation is stabilized by an effective ‘salt-bridge’, i.e., Gly H₃N⁺… COO⁻ Arg interaction. Surprisingly, in both the families the unusual folded side-chain dispositions of the Gln residue favor the formation of a unique intra-residue ‘main-chain to side-chain’ H-bond, i.e., N–H…N^ε interaction, encompassing a seven-membered ring motif. The conformational attributes may be valuable in de novo construction of structure-based drug candidates having sufficient stimulating activity.     

On intrinsic equivalences of the finite helical axis,the instantaneous helical axis,and the SARA approach. A mathematical perspective

Accurate determination of joint axes is essential for understanding musculoskeletal function. Whilst numerous algorithms to compute such axes exist, the conditions under which each of the methods performs best remain largely unknown. Typically, algorithms are evaluated for specific conditions only limiting the external validity of conclusions regarding their performance. We derive exact mathematical relationships between three commonly used algorithms for computing joint axes from motion data: finite helical axes (FHA), instantaneous helical axes (IHA) and SARA (symmetrical axis of rotation approach), including relationships for an extension to the mean helical axes methods that facilitate determining joint centres and axes. Through the derivation of a sound mathematical framework to objectively compare the algorithms we demonstrate that the FHA and SARA approach are equivalent for the analysis of two time frames. Moreover, we show that the position of a helical axis derived from the IHA using positional data is affected by a systematic error perpendicular to the true axis direction, whereas the axis direction is identical to those computed with either the FHA or SARA approach (true direction). Finally, with an appropriate choice of weighting factors the mean FHA (MFHA) method is equivalent to the Symmetrical Centre of Rotation Estimation (SCoRE) algorithm for determination of a Centre of Rotation (CoR), and similarly, equivalent to the SARA algorithm for determination of an Axis of Rotation (AoR). The deep understanding of the equivalences between methods presented here enables readers to choose numerically efficient, robust methods for determining AoRs and CoRs with confidence.     

Participation of the microsomal electron transport system involving cytochrome P-450 in ω-oxidation of fatty acids

The participation of the microsomal electron transport system involving cytochrome P-450 in ω-oxidation of fatty acids by a rat liver preparation was examined since ω-oxidation involves microsomal reactions requiring both NADPH and molecular oxygen.

ω-Oxidation of fatty acids was inhibited by CO and by the antibody against NADPH-cytochrome c reductase. The addition to the reaction mixture of drugs which interact with cytochrome P-450 inhibited ω-oxidation. It is concluded that the microsomal electron transport system involving cytochrome P-450 functions in ω-oxidation of fatty acids.     

Malat-dehydrogenase isoenzymbanden als potentielles chemotaxonomisches kriterium für cyanophyceen-species

Malate dehydrogenase isoenzymes from eight cyanophycean species were investigated with polyacrylamide disc gel electrophoresis. Using 7% acrylamide the pherograms from each species showed 5–8 zones with malate dehydrogenase activity. It was demonstrated that in Anabaena flos-aquae there are 8 isoenzyme bands which include 3 forms of equal molecular weight, two of which consist of several isomers differing in their net charge. The MDH zymograms of the blue-green algae investigated can be used as “fingerprints”. The isoenzyme pattern of the MDHs of Anacystis nidulans makes its position in the order Chroococcales uncertain.

Résumé

Wäßrige Extrakte aus acht Cyanophyceenarten wurden einer Polyacrylamid-Discelektrophorese unterzogen und die erhaltenen Elektropherogramme auf Malat-Dehydrogenase (MDH)-Aktivität geprüft. Dabei ergab sich, daß unter unseren Versuchsbedingungen die MDH der getesteten Cyanophyceen in 5–8 Isoenzymbanden aufspaltet. Für Anabaena flos-aquae konnte gezeigt warden, daß sich die 8 Isoenzymbanden auf wenigstens 3 Molekulargewichts-Isomere zurückführen lassen, von denen zwei noch mehrere Ladungsisomere bilden. Die erhaltenen Zymogramme zeigen “fingerprint”-Charakter. was ihre mögliche Verwendbarkeit für die Chemotaxonomie der Cyanophyta nahelegt. Die Stellung von Anacystis nidulans innerhalb der CyanophyceenOrdnungen wird diskutiert.     

Variation in δC values for the seagrass Thalassia testudinum and its relations to mangrove carbon

Carbon isotope ratios (¹³C/¹²C) were measured for the leaves of the seagrass Thalassia testudinum Banks ex König and carbonates of shells collected at the seagrass beds from seven sites along the coast of southern Florida, U.S.A. The δ¹³C values of seagrass leaves ranged from −7.3 to −16.3‰ among different study sites, with a significantly lower mean value for seagrass leaves from those sites near mangrove forests (−12.8 ± 1.1‰) than those far from mangrove forests (−8.3 ± 0.9‰; P < 0.05). Furthermore, seagrass leaves from a shallow water area had significantly lower δ¹³C values than those found in a deep water area (P < 0.01). There was no significant variation in δ¹³C values between young and mature leaves (P = 0.59) or between the tip and base of a leaf blade (P = 0.46). Carbonates of shells also showed a significantly lower mean δ¹³C value in the mangrove areas (−2.3 ± 0.6‰) than in the non-mangrove areas (0.6 ± 0.3‰; P <0.025). In addition, the δ¹³C values of seagrass leaves were significantly correlated with those of shell carbonates (δ¹³C seagrass leaf = −9.1 + 1.3δ¹³C shell carbonate (R² = 0.83, P < 0.01)). These results indicated that the input of carbon dioxide from the mineralization of mangrove detritus caused the variation in carbon isotope ratios of seagrass leaves among different sites in this study.     

Binding of the C6-zinc cluster protein, AFLR, to the promoters of aflatoxin pathway biosynthesis genes in Aspergillus parasiticus

AFLR is a Zn₂Cys₆-type sequence-specific DNA-binding protein that is thought to be necessary for expression of most of the genes in the aflatoxin pathway gene cluster in Aspergillus parasiticus and A. flavus, and the sterigmatocystin gene cluster in A. nidulans. However, it was not known whether AFLR bound to the promoter regions of each of the genes in the cluster. Recently, A. nidulans AFLR was shown to bind to the motif 5′-TCGN₅CGA-3′. In the present study, we examined the binding of AFLR to promoter regions of 11 genes in the A. parasiticus cluster. Based on electrophoretic mobility shift assays, the genes nor1, pksA, adhA, norA, ver1, omtA, ordA, and, vbs, had at least one 5′-TCGN₅CGA-3′ binding site within 200 bp of the translation start site, and pksA and ver1 had an additional binding site further upstream. Although the promoter region of avnA lacked this motif, AFLR bound weakly to the sequence 5′-TCGCAGCCCGG-3′ at −110 bp. One region in the promoter of the divergently transcribed genes aflR/aflJ bound weakly to AFLR even though it contained a site with at most only 7 bp of the 5′-TCGN₅CGA-3′ motif. This partial site may be recognized by a monomeric form of AFLR. Based on a comparison of 16 possible sites, the preferred binding sequence was 5′-TCGSWNNSCGR-3′.     

Characteristics of microbial fermentation and potential digestibility of fiber in the hindgut of dugongs (Dugong dugon)

The number of microorganisms in the hindgut of dugongs (Dugong dugon) were estimated and their in vitro volatile fatty acid (VFA) production and degradation of eelgrass measured. Scanning electron microscopy showed that some rod bacteria attached to the surface of plant tissue degraded and eroded the cell walls. Number of starch-, lactate-, cellobiose-, pectin-, xylan- and cellulose-utilizing bacteria, sulfate-reducing bacteria and methane-producing bacteria were estimated at 10⁹ ∼ 10¹⁰ colony forming units g^-1. Microorganisms degraded the cellulose and noncellulolytic components of the eelgrass, and about 47.3% of dry matter was degraded after 36 h in vitro incubation. The total VFA concentration was 10.5 mmol dL^-1 at 36 h incubation, which included 55.7 mol% acetate, 18.0 mol% n-butyrate and 15.1 mol% propionate. The gas composition of in vitro fermentation was 68.4% carbon dioxide, 22.2% methane and 9.4% hydrogen.     

Cytochrome c interaction with the mitochondrial membrane: A spin label study

Horse-heart ferrocytochrome c has been labeled with N-(2,2,5,5-tetramethyl-3-pyrrolidinyl-1-oxyl) iodoacetamide at methionine-65. The paramagnetic resonance spectrum of labeled ferricytochrome c indicates a weak immobilization of the radical (τ_c = 9.3·10⁻¹⁰ sec) which becomes stronger upon binding of labeled cytochrome c to cytochrome c-depleted mitochondrial membranes (τ_c = 3.3·10⁻⁹ sec). The hyperfine coupling constant remains, however, unchanged (16.7 ± 0.1 gauss) indicating that the cytochrome c binding site is highly polar. The region where cytochrome c is bound to the membrane is insensitive to large variations of medium viscosity.