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R Mendelsohn

Publications and source records attributed to R Mendelsohn.

At least 37 records · Page 2Linked to original sources
PubMed1999

Stability of annexin V in ternary complexes with Ca2+ and anionic phospholipids: IR studies of monolayer and bulk phases.

Annexin V (AxV) is a member of a family of proteins that exhibit functionally relevant Ca2+-dependent binding to anionic phospholipid membranes. Protein structure and stability as a function of Ca2+ and phospholipids was studied by bulk phase infrared (IR) spectroscopy and by IR reflection-absorption spectroscopy (IRRAS) of monolayers in situ at the air/water (A/W) interface. Bulk phase experiments revealed that AxV undergoes an irreversible thermal denaturation at approximately 45-50 degreesC, as shown by the appearance of amide I bands at 1617 and 1682 cm-1. However, some native secondary structure is retained, even at 60 degreesC, consistent with a partially unfolded "molten globule" state. Formation of the Ca2+/phospholipid/protein ternary complex significantly protects the protein from thermal denaturation as compared to AxV alone, Ca2+/AxV, or lipid/AxV mixtures. Stabilization of AxV secondary structure by a DMPA monolayer in the presence of Ca2+ was also observed by IRRAS. Spectra of an adsorbed AxV film in the presence or absence of Ca2+ showed a 10 cm-1 shift in the amide I mode, corresponding to loss of ordered structure at the A/W interface. In both the bulk phase and IRRAS experiments, protection against H-->D exchange in AxV was enhanced only in the ternary complex. The combined data suggest that the secondary structure of AxV is strongly affected by the Ca2+/membrane component of the ternary complex whereas lipid conformational order is unchanged by protein.

Animals↗
PubMed1999

Some relationships between membrane phospholipid domains, conformational order, and cell shape in intact human erythrocytes.

A novel method developed in this laboratory [D.J. Moore et al., Biochemistry 35 (1996) 229-235; D.J. Moore et al., Biochemistry 36 (1997) 660-664] to study the conformational order and the propensity for domain formation of specific phospholipids in intact human erythrocytes is extended to two additional species. Acyl chain perdeuterated 1,2-dilauroylphosphatidylethanolamine (diC12PE-d46) was incorporated preferentially (in separate experiments) into the inner leaflet of stomatocytic erythrocytes and into the outer leaflet of echinocytic erythrocytes, while acyl chain perdeuterated 1,2-dipentadecanoylphosphatidylcholine (diC15PC-d58) was incorporated into the outer leaflet of echinocytic erythrocytes. The conformational order and phase behavior of the incorporated molecules were monitored through FT-IR studies of the temperature dependence of the CD2 stretching vibrations. For both diC12PE-d46 and diC15PC-d58, the gel-->liquid crystal phase transition persisted when these lipids were located in the outer leaflet of echinocytic cells, a result indicative of the persistence of phospholipid domains. In each case, the transition widths were broadened compared to the pure lipids, suggestive of either small domains or the presence of additional molecular components within the domains. The conformational order of diC12PE-d46 differed markedly depending on its location and the morphology of the cells. When located predominantly in the inner membrane of stomatocytes, the phase transition of this species was abolished and the conformational order compared with pure lipid vesicles at the same temperature was much lower. The current results along with our previous studies provide a sufficient experimental basis to deduce some general principles of phospholipid conformational order and organization in both normal and shape-altered erythrocytes.

Cell Size↗
PubMed1999

Complementary information on bone ultrastructure from scanning small angle X-ray scattering and Fourier-transform infrared microspectroscopy.

Scanning small angle X-ray scattering (scanning SAXS) and Fourier-transform infrared microspectroscopy (FT-IRM) have previously been utilized independently to characterize the structural properties of bone in an anatomical position-resolved fashion. Whereas SAXS provides a direct measure of the physical characteristics of apatitic crystals, FT-IRM assesses structure of both mineral and organic matrix at the molecular level. In the present study both methods were applied to examine the same developing bone tissue from the L-4 vertebra of a 14-month-old (accidental death). A 200-microm-thick section was processed for examination by scanning electron microscopy and SAXS. Spectra were collected at 200 microm spatial resolution at specific locations in cortical and cancellous bone. Parameters determined included total SAXS intensity, crystal thickness (T), and degree and direction of predominant crystal orientation. For FT-IRM analysis, a section 4 microm thick was cut longitudinally from the top of the sample. Spectra of regions 100 x 100 microm2 were acquired from the same locations as the SAXS spectra. Integrated areas of the phosphate nu(1,3) collagen amide I, and carbonate nu2 absorbances, were calculated to obtain mineral: matrix and carbonate:mineral ratios. The relative quantities of types A, B, and labile carbonate (substituted for apatite hydroxyl, phosphate, and surface positions, respectively) were also evaluated. Polarized FT-IRM data were collected to determine molecular orientation of the apatite and collagen components. The results of this study show that the information obtained from the two techniques is complementary. Both SAXS and FT-IRM data revealed that the crystals were significantly larger in the cancellous region compared with the cortical region, that mineralization was greater in the cortex, and that the crystals were oriented to a larger degree in the cancellous compared with the cortical bone. The scanning SAXS measure of crystal thickness was significantly correlated to the FT-IRM measures of crystallinity, type A carbonate substitution, and crystal orientation. In conclusion, it was found that the combined use of SAXS and FT-IRM provides valuable, unique information on structural changes in bone at both the microstructural and ultrastructural level. Although each method can be used individually, the combination of techniques provides additional insights into the mechanism of bone crystal maturation.

Apatites↗
PubMed1998

Domain structure and molecular conformation in annexin V/1,2-dimyristoyl-sn-glycero-3-phosphate/Ca2+ aqueous monolayers: a Brewster angle microscopy/infrared reflection-absorption spectroscopy study.

Annexins comprise a family of proteins that exhibit a Ca2+-dependent binding to phospholipid membranes that is possibly relevant to their in vivo function. Although substantial structural information about the ternary (protein/lipid/Ca2+) interaction in bulk phases has been derived from a variety of techniques, little is known about the temporal and spatial organization of ternary monolayer films. The effect of Ca2+ on the interactions between annexin V (AxV) and anionic DMPA monolayers was therefore investigated using three complementary approaches: surface pressure measurements, infrared reflection-absorption spectroscopy (IRRAS), and Brewster angle microscopy (BAM). In the absence of Ca2+, the injection of AxV into an aqueous subphase beneath a DMPA monolayer initially in a liquid expanded phase produced BAM images revealing domains of protein presumably surrounded by liquid-expanded lipid. The protein-rich areas expanded with time, resulting in reduction of the area available to the DMPA and, eventually, in the formation of condensed lipid domains in spatial regions separate from the protein film. There was thus no evidence for a specific binary AxV/lipid interaction. In contrast, injection of AxV/Ca2+ at a total Ca2+ concentration of 10 microM beneath a DMPA monolayer revealed no pure protein domains, but rather the slow formation of pinhead structures. This was followed by slow (>2 h) rigidification of the whole film accompanied by an increase in surface pressure, and connection of solid domains to form a structure resembling strings of pearls. These changes were characteristic of this specific ternary interaction. Acyl chain conformational order of the DMPA, as measured by nu(sym)CH2 near 2850 cm(-1), was increased in both the AxV/DMPA and AxV/DMPA/Ca2+ monolayers compared to either DMPA monolayers alone or in the presence of Ca2+. The utility of the combined structural and temporal information derived from these three complementary techniques for the study of monolayers in situ at the air/water interface is evident from this work.

Annexin A5↗
PubMed1998

Infrared microspectroscopic imaging of biomineralized tissues using a mercury-cadmium-telluride focal-plane array detector.

A 64 x 64 mercury-cadmium-telluride focal-plane array detector attached to a Fourier transform infrared microscope was used to spectroscopically image 5 microm sections of canine alveolar bone tissue in the fingerprint region of the infrared spectrum. By ratioing the relative intensities of specific bands across the images, it is possible to obtain spatial distributions of the mineral-to-matrix ratio and mineral maturity as a function of distance from an osteon.

Animals↗
PubMed1997

Adipocyte fatty acid-binding protein: interaction with phospholipid membranes and thermal stability studied by FTIR spectroscopy.

Fatty acid-binding proteins (FABPs) found in many tissues constitute a family of low molecular weight proteins that are suggested to function as intracellular transporters of fatty acids. Studies of the transfer kinetics of fluorescent anthroyloxy-labeled long-chain fatty acids from FABP to model membranes led to the suggestion that the FABPs, typically considered to be cytosolic proteins, could nevertheless interact directly with membranes [Wootan, M. G., et al. (1993) Biochemistry 32, 8622-8627]. In the current study, the interaction of the adipocyte FABP (A-FABP) with vesicles of various phospholipids has been directly measured and confirmed with FTIR spectroscopy. The strength of this interaction was inferred from the lowering of the gel-liquid-crystal phase transition temperature as monitored from temperature-induced variations in the acyl chain CH2 stretching frequencies and from the intensities of the components of the CH2 wagging progressions. A-FABP interacts more strongly with anionic phospholipids (phosphatidylserine and cardiolipin) than with zwitterionic phosphatidylcholine. Unsaturation in the acyl chains leads to a greater reduction in Tm (stronger lipid-protein interaction). In contrast, neutralization of A-FABP surface charges by acetylation considerably weakens the interaction. Comparison of the shifts in lipid melting temperatures with those induced by other proteins suggests that A-FABP behaves like a typical peripheral membrane protein. The degree of membrane interaction correlates directly with the rate of fatty acid transfer, suggesting that contact between A-FABP and membranes is functionally related to its fatty acid transport properties. As expected, the protein exhibits a predominantly beta-sheet structure. It was found to aggregate with increasing temperature. With the exception of minor differences between the pure and lipid-associated A-FABP in the 1640-1660 cm-1 region, both the protein structure and thermal stability appeared essentially unchanged upon interaction with the lipid.

Acetylation↗
PubMed1997

Lipid domains and orthorhombic phases in model stratum corneum: evidence from Fourier transform infrared spectroscopy studies.

A three component model for the lipid barrier of the stratum corneum (SC) consisting of ceramide III, cholesterol, and perdeuterated palmitic acid, has been characterized by Fourier transform infrared spectroscopy. At physiological temperature the CD2 scissoring mode of the palmitic acid methylenes, and the CH2 rocking mode of the ceramide methylenes, are each split into two components. This indicates that both components exist in separate, conformationally ordered phases, probably with orthorhombic perpendicular subcells. The magnitude of the splitting indicates that the domains are at least 100 chains in size. The thermotropic behavior of the CD2 stretching vibrations demonstrates that conformational disordering of the palmitic acid commences at 42 degrees C with a transition midpoint of 50 degrees C. The CH2 stretching frequency indicates the ceramide chains remain ordered until 50 degrees C then disorder with a midpoint of 67 degrees C. The results provide a molecular characterization for the complex low temperature (10-40 degrees C) dynamic behavior suggested by recent 2H NMR experiments.

Biophysical Phenomena↗
PubMed1997

Conformational order of specific phospholipids in human erythrocytes: correlations with changes in cell shape.

Acyl chain perdeuterated dimyristoylphosphatidylcholine (DMPC-d54) and dimyristoyphosphatidylserine (DMPS-d54) were incorporated into human erythrocytes. Light microscopy demonstrated that erythrocytes incubated with an equimolar mixture of DMPC-d54/DMPS or DMPC/DMPS-d54 remained mostly discocytic whereas cells incubated with either DMPC-d54 or DMPS-d54 alone became echinocytic or stomatocytic, respectively. Cells in which the aminophospholipid translocating protein was inhibited became echinocytic when incubated with DMPS-d54. Fourier transform infrared (FTIR) spectroscopy was used to monitor conformational order in the acyl chains of the incorporated phospholipid, as detected through the asymmetric CD2 stretching vibrations in the intact cells. In cells incubated with equimolar mixtures of DMPC-d54/DMPS or DMPC/DMPS-d54, the deuterated species exhibited no thermotropic phase transitions but revealed chain order intermediate between the gel and liquid-crystal states. In contrast, DMPS-d54 incorporated into the outer leaflet of echinocytic erythrocytes was conformationally ordered while the same species incorporated into the inner leaflet of stomatocytic erythrocytes was highly disordered at all temperatures studied. Finally, DMPC-d54 incorporated into the outer leaflet of echinocytic erythrocytes exhibited a phase transition, suggesting that this species persists in domains. These data indicate that the acyl chain conformational order of specific phospholipids in the intact human erythrocyte is changed with alterations in cell morphology.

Dimyristoylphosphatidylcholine↗
PubMed1997

FTIR microspectroscopic analysis of normal human cortical and trabecular bone.

Fourier transform infrared microspectroscopy (FTIRM) has been used to study the changes in mineral and matrix content and composition in replicate biopsies of nonosteoporotic human cortical and trabecular bone. Changes in osteonal bone in these same samples were reported previously. Spectral maps along and across the lamellae were obtained from iliac crest biopsies of two necropsy cases. Mineral:matrix ratios, calculated from the integrated areas of the phosphate nu1, nu3 band at 900-1200 cm-1 and the amide I band at approximately 1585-1725 cm-1, respectively, were relatively constant in both directions of analysis, i.e., along and across the lamellae. Analysis of the components of the nu1, nu3 phosphate band with a combination of second-derivative spectroscopy and curve fitting revealed the presence of 11 major underlying moieties. Of these, the ratio of the relative areas of the two underlying bands at approximately 1020 and approximately 1030 cm-1 has been shown to be a sensitive index of variation in crystal perfection in both human osteonal bone and in synthetic, poorly crystalline apatites. This ratio was calculated in both cortical and trabecular bone from human iliac crest biopsies along and across the lamellae. The ratio decreased, going from the periosteum to the medullary cavity in the cortical bone, and from the periphery towards the center of trabeculae. These observations were consistent within serial sections obtained from the same biopsy, multiple biopsies obtained from the same necropsy specimen, and biopsies obtained from the two different necropsy specimens. The results presented here along with previously reported changes in osteonal bone show a relation between bone age and "crystallinity/maturity" (a parameter dependent on crystallite size, hydroxyapatite-like stoichiometry, abundance of substituting ions such as CO32-; the more crystalline/mature, the more hydroxyapatite-like stoichiometry, the bigger the crystallite size, the less the ion substitution by ions such as CO32-) as deduced by the 1020/1030 cm-1 ratio. Invariably, younger normal bone is less mature/crystalline than older. These results provide a "baseline" for description of mineral properties, to which diseased bones may be compared.

Aged↗
PubMed1997

FTIR microspectroscopic analysis of human iliac crest biopsies from untreated osteoporotic bone.

Historically, osteoporosis has been defined as a disease in which there is "too little bone, but what there is, is normal." As a result of research design and sample selection limitations, published data contradict and confirm the historical definition. Because of these limitations, it has been hard to assess the contribution of mineral quality to mechanical properties, and to select therapeutic protocols that optimize bone mineral properties. The coupling of an optical microscope to an infrared spectrometer enables the acquisition of spectral data at known sites in a histologic section of mineralized tissue without loss of topography and/or orientation. The use of second-derivative spectroscopy coupled with curve-fitting techniques allows the qualitative and quantitative assessment of mineral quality (crystallite size and perfection, mineral:matrix ratio) at well-defined morphologic locations. We have previously applied these techniques to the study of normal human osteonal, cortical, and trabecular bone. The results indicated that the newly deposited bone mineral is less "crystalline/mature" than the older one. In the present study, Fourier transform infrared microspectroscopy (FTIRM) was applied to the study of human osteonal and cortical bone from iliac crest biopsies of untreated osteoporotic patients. The hypothesis tested was that osteoporotic bone mineral is monotonically different in its properties expressed as "crystallinity/maturity" than the normal. The results indicate significant differences in the mineral properties as expressed by crystal size and perfection, with the mineral from osteoporotic bone being more crystalline/mature than the normal.

Adult↗
PubMed1997

Structure and orientation of lung surfactant SP-C and L-alpha-dipalmitoylphosphatidylcholine in aqueous monolayers.

SP-C, a pulmonary surfactant-specific protein, aids the spreading of the main surfactant phospholipid L-alpha-dipalmitoylphosphatidylcholine (DPPC) across air/water interfaces, a process that has possible implications for in vivo function. To understand the molecular mechanism of this process, we have used external infrared reflection-absorption spectroscopy (IRRAS) to determine DPPC acyl chain conformation and orientation as well as SP-C secondary structure and helix tilt angle in mixed DPPC/SP-C monolayers in situ at the air/water interface. The SP-C helix tilt angle changed from approximately 24 degrees to the interface normal in lipid bilayers to approximately 70 degrees in the mixed monolayer films, whereas the acyl chain tilt angle of DPPC decreased from approximately 26 degrees in pure lipid monolayers (comparable to bilayers) to approximately 10 degrees in the mixed monolayer films. The protein acts as a "hydrophobic lever" by maximizing its interactions with the lipid acyl chains while simultaneously permitting the lipids to remain conformationally ordered. In addition to providing a reasonable molecular mechanism for protein-aided spreading of ordered lipids, these measurements constitute the first quantitative determination of SP-C orientation in Langmuir films, a paradigm widely used to simulate processes at the air/alveolar interface.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1996

Conformational disorder in unsaturated phospholipids by FTIR spectroscopy.

Conformational disorder in liquid alkenes and in the L alpha and Hparallel phases of some unsaturated phospholipids has been monitored by FTIR spectroscopy. The CH2 wagging region (1330-1390 cm-1) in saturated chains contains vibrations of particular 2- and 3-bond conformational states as follows: 1341 cm-1, end-gauche (eg); 1352 cm-1, double gauche (gg); 1368 cm-1, the sum of kink and gtg states. In unsaturated chains, this spectral region revealed an additional band at 1362 cm-1 and (occasionally) a feature near 1348 cm-1. The 1362 cm-1 band is tentatively assigned to the wagging of CH2 groups adjacent to the C = C bond. Substantial populations of both gg and (kink+gtg) states are evident in the L alpha phases of unsaturated phosphatidylcholines (PC's). Unsaturated phosphatidylethanolamines (PE's) are more ordered than their PC counterparts, and possess fewer gg and eg states. Chain disorder in the Hparallel phase of PE's approaches that in L alpha phases of unsaturated PC's. Changes in conformer distributions during the L alpha-->Hparallel transition in 1,2-dioleoylphosphatidylethanolamine (DOPE), 1-palmitoyl,2-oleoylphosphatidylethanolamine (POPE), 1,2-dielaidoylphosphatidylethanolamine (DEPE), and N-methyl-DOPE(N-MeDOPE) were semi-quantitatively estimated. For DOPE and DEPE, slight cooperative increases in both gg and (kink+gtg) states occur, for POPE only the gg population increases and for N-MeDOPE only the kink+gtg populations increase. These disorder increases are consistent with the small calorimetric delta H for this transition. Difficulties in quantitative determination of conformational disorder in unsaturated chains are discussed.

Alkenes↗
PubMed1996

Three of four cysteines, including that responsible for substrate activation, are ionized at pH 6.0 in yeast pyruvate decarboxylase: evidence from Fourier transform infrared and isoelectric focusing studies.

Oligonucleotide-directed site-specific mutagenesis was carried out on pyruvate decarboxylase (EC 4.1.1.1) from Saccharomyces cerevisiae at three of the four cysteines (152, 221, and 222), the fourth (69) being buried according to X-ray crystallographic results [Arjunan et al. (1996) J. Mol. Biol. 256, 590-600]. All of the variants still retained significant activity, and all could be purified to homogeneity. FT-IR experiments were run on the C221S, C222S, C221S/C222S and C152A variants, as well as on the wild-type enzyme. There is a band present at 2557 cm-1 in the spectra of all variants and the wild-type enzyme, except in the spectrum of the C152A variant. This frequency is appropriate to a cysteine S-H stretching mode. It was therefore concluded that C152 is the only undissociated cysteine on the enzyme at pH 6.0, the pH optimum of this enzyme, whereas C221, C222, and C69 are all ionized. Isoelectric focusing experiments were carried out on all of these variants, as well as on the H92A variant (H92 is across the domain divide on the alpha domain, from C221 located on the beta domain). The variation in isoelectric points deduced from the data was consistent with removal of negative charges concomitant with the C221S, C222S, and C221S/C222S substitutions and removal of a positive charge with the H92A substitution when compared to that of the wild-type enzyme. The results of these two types of experiments are in good accord and suggest that the site of substrate activation at C221 [Baburina et al. (1994) Biochemistry 33, 5630-5635] is comprised of a Cys221S- +HHis92 ion pair, not unlike that found in papain and glyceraldehyde-3-phosphate dehydrogenase. This finding suggests that the regulatory site of this enzyme has been optimized for nucleophilic reactivity between the thiolate of C221 and the keto carbon of the 2-oxoacid.

Cysteine↗
PubMed1996

Fluctuations in IR spectral parameters detected in mixed acyl chain membranes of Acholeplasma laidlawii B.

Acholeplasma laidlawii B cells were grown at 37 degrees C on three binary C16:0-d(31)/C18:1 fatty acid mixtures at initial mol ratios of 3:2, 1:1, and 2:3. These mol ratios produced final C16:0-d(31)/C18:1 lipid acyl chain mol ratios of 1.66 +/- 0.23 (n=6), 1.3 +/- 0.20 (n=6) and 0.58 +/- 0.09 (n=10), respectively, in the membrane of the microorganism. Membrane conformational order for the deuterated and proteated acyl chains in intact cells was monitored by FT-IR spectroscopy through the thermotropic response of the acyl chain CD2 and CH2 stretching frequencies. Intact cells and isolated membranes revealed broad phase transitions centered well below the growth temperature. This result differs from previous studies (Moore, D.J. and Mendelsohn, R. (1994) Biochemistry 33, 4080-4085) of cells grown on a single saturated fatty acid source, where Tm was close to the growth temperature. Fluctuations in IR spectral parameters from the liquid crystalline phases were detected in ten separate samples of cells grown on a 2:3 mixture (final mol ratio 0.58:1) of C16:0-d(31)/C18:1, and in no other cell preparation. These were manifest by reduced precision in the measurement of CH2 and CD2 stretching frequencies and are attributed to fluctuations in the membrane conformational order. In addition to conformational order fluctuations in intact cells, similar behavior was noted for the simple binary phosphatidylcholine (PC) mixture, DOPC/1-C16:0-d(31),2-C18:1 PC (2:1 molar ratio). In this instance, the fluctuations were also detected through the temporal and thermotropic response of the relative intensity of the 1341 cm(-1) band assigned to end-gauche conformers about the penultimate C-C bond in the oleoyl chains. The relationship of these observations to the Raman spectroscopic detection of packing fluctuations in highly unsaturated PC's (Litman, B.J., Lewis, N., and Levin, I.W. (1991) Biochemistry 30, 313-319) is considered.

Acholeplasma laidlawii↗
PubMed1996

Conformational order of phospholipids incorporated into human erythrocytes: an FTIR spectroscopy study.

Acyl chain perdeuterated dimyristoylphosphatidylcholine (DMPC-d54) and dimyristoylphosphatidylserine (DMPS-d54) were incorporated by incubation into human erythrocytes. Light microscopic analysis demonstrated that erythrocytes incubated with DMPC-d54 became echinocytic while those incubated with DMPS-d54 became stomatocytic. This indicates that DMPC-d54 was incorporated preferentially into the outer monolayer whereas DMPS-d54 was selectively incorporated into the inner monolayer. Fourier transform infrared (FTIR) spectroscopy was used to monitor the conformational order of the incorporated phospholipids. The asymmetric CD2 stretching frequency of the inserted perdeuterated acyl chains was measured in both isolated membranes and intact erythrocytes as a function of temperature. DMPC-d54 incorporated into erythrocytes exhibited a cooperative phase transition at approximately 19 degrees C, i.e., at the same temperature as pure vesicles. In contrast, DMPS-d54 incorporated into red cells exhibited no phase transition, but possessed conformational order similar to that of the liquid-crystalline state. These results suggest that DMPC-d54 persists in domains in the outer monolayer while DMPS-d54 is dispersed in the inner monolayer. These experiments are the first to demonstrate that FTIR spectroscopy can be utilized to monitor directly a specific species of lipid molecule from the entire phospholipid population.

Adult↗
PubMed1996

FTIR microspectroscopic analysis of human osteonal bone.

Fourier Transform Infrared Microspectroscopy (FTIRM) has been used to study the changes in mineral and matrix content and composition in replicate biopsies of nonosteoporotic human osteonal bone. Spectral maps in four orthogonal directions (in 10 microm steps) from the centers towards the peripheries of individual osteons were obtained from iliac crest biopsies of two necropsy cases. Mineral to matrix ratios, calculated from the ratio of integrated areas of the phosphate nu1,nu3 band at 900-1200 cm-1 to the amide I band at 1585-1725 cm-1, increased from the center to the periphery of the osteon. The total carbonate (based on the nu2 band at approximately 850-900 cm-1) to phosphate nu1,nu3 ratio decreased as the mineral to matrix ratio increased. Analysis of the nu2 CO32- band with a combination of second-derivative spectroscopy and curve fitting revealed a decrease in "labile" carbonate, a slight decrease in Type A and a slight increase in Type B carbonate from the center to the periphery of the osteon. Similar analysis of the components of the nu1,nu3 phosphate band with a combination of second-derivative spectroscopy and curve fitting revealed the presence of 11 major underlying moieties. These components were assigned by comparison with published frequencies for apatite and acid-phosphate containing calcium phosphates. The most consistent variations were alterations in the relative percent areas of bands at approximately 1020 and approximately 1030 cm-1, which had previously been assigned to nonstoichiometric and stoichiometric apatites, respectively. This ratio was used as an index of variation in crystal perfection throughout the osteon. This ratio decreased as the mineral to matrix ratio increased. The reproducibility of these parameters at multiple sites in multiple biopsies suggests their applicability for the analysis of mineral changes in disease.

Aged↗