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1.
Difference spectra between resting and excited nerve in the infrared region between 2000 and 1000 cm-1 have been examined with a resolution of 0.5 cm-1. Spectra were obtained with a modified Perkin-Elmer model 521 grating infrared spectrophotometer (Perkin-Elmer Corp., Instrument Div., Norwalk, Conn.), and the signal-to-noise (S/N) ratio was improved by time averaging and digital smoothing. Peaks occurring in the regions around 1030 and 1066 cm-1 are identified as P-O-C stretch, at 1410-1414 cm-1 as C-H deformation, and at 1750 cm-1 as carbonyl stretch. The difference peaks appear to be due to a shift of about 1 cm-1 in the absorption band to lower frequencies for the three lower frequency bands and to a higher frequency for 1750 cm-1 band. Since the difference peaks appear when the nerve is modulated by a propagated action potential it is concluded that the changing electrical field across the nerve membrane is perturbing the absorption spectrum. From evidence presented it appears likely that these difference peaks are due to phospholipids in the nerve membrane and that they may be related to conformational changes associated with membrane permeability.  相似文献   

2.
Several polypeptides have been found to adopt an unusual domain structure known as the parallel beta-helix. These domains are characterized by parallel beta-strands, three of which form a single parallel beta-helix coil, and lead to long, extended beta-sheets. We have used ATR-FTIR (attenuated total reflectance-fourier transform infrared spectroscopy) to analyze the secondary structure of representative examples of this class of protein. Because the three-dimensional structures of parallel beta-helix proteins are unique, we initiated this study to determine if there was a corresponding unique FTIR signal associated with the parallel beta-helix conformation. Analysis of the amide I region, emanating from the carbonyl stretch vibration, reveals a strong absorbance band at 1638 cm(-1) in each of the parallel beta-helix proteins. This band is assigned to the parallel beta-sheet structure. However, components at this frequency are also commonly observed for beta-sheets in many classes of globular proteins. Thus we conclude that there is no unique infrared signature for parallel beta-helix structure. Additional contributions in the 1638 cm(-1) region, and at lower frequencies, were ascribed to hydrogen bonding between the coils in the loop/turn regions and amide side-chain interactions, respectively. A 13-residue peptide that forms fibrils and has been proposed to form beta-helical structure was also examined, and its FTIR spectrum was compared to that of the parallel beta-helix proteins.  相似文献   

3.
本研究的目的是以性腺完整的动物为模型调查左乙拉西坦(LEV)是否对于骨矿物质密度、骨结构和骨代谢生化指标产生影响,探究抗癫痫药物左乙拉西坦(LEV)是否对骨骼健康存在显著风险。本研究将实验大鼠分为对照组和试验组,每组10只。对照大鼠接受标准实验室饮食(SLD),而试验组中的大鼠喂食富含LEV的实验室饮食12周,并以双能X-线吸收仪测量全身、股骨和腰椎的骨密度。在骨组织中检查骨标记的浓度,股骨和胫骨均用于生物力学测试。研究结果表明:在LEV组中,脂肪组织的绝对值和相对值显著降低,全身骨密度增高,骨碱性磷酸酶(BALP)、Ⅰ型胶原C末端肽(CTX-1)和Ⅰ型前胶原氨基端前肽(PINP)浓度显著增加。本研究初步得出结论:在性腺完整的大鼠模型中长期施用LEV对骨质不具有负面影响。骨矿物质密度(BMD)的显著增加可能表明LEV对骨质可能有正面影响。  相似文献   

4.
Infrared bands for CO bound to mitochondria from bovine and porcine hearts, bovine brain, rat kidney, and blowfly flight muscle and to intact blowfly flight muscle have been measured in the carbon-oxygen stretch region. Each spectrum contains a narrow band near 1963 cm-1 similar to the major band found earlier for the carbonyl cytochrome c oxidase purified from bovine heart. A second band near 1959 cm-1 ascribed to a less stable conformer of the purified oxidase carbonyl is also detected in mitochondria. These spectra support very similar CO (and O2) binding sites among all the oxidases examined whether the enzyme is purified or is still within mitochondria or intact tissue and therefore suggest that the reduced heme A ligand binding site has been highly conserved during evolution.  相似文献   

5.
The infrared spectra for carbon monoxide complexed to hemoglobins were examined in the C-O stretch region. Deconvolution of the spectra requires four bands and supports the presence of four distinct conformers at the ligand binding site. Most typical hemoglobins exhibit only one predominant conformer for each subunit represented by a band at 1951 cm-1 in contrast to myoglobins, which typically exist in two major conformations. Several hemoglobins with an enlarged heme pocket are shown to shift the C-O frequency into the higher frequency conformer regions. Many factors, including pH, temperature, solvents, and divalent metals, are also shown to be capable of expanding the heme pocket. Only very specific structural changes that can reduce the size of the heme pocket will result in the lower frequency conformers. The weighted averages of the multiple CO vibrational frequencies are linearly related to the single 13CO NMR chemical shift values and to the exponential of fast CO on-rates. Conformer interconversion occurs at a rate greater than 10(4) s-1. The infrared C-O stretch spectra provide qualitative and quantitative information on the structural dynamics, stability, and ligand binding properties of hemoglobins.  相似文献   

6.
Effect of modulated pulsed electromagnetic fields (PEMFs; carrier frequency, 14 MHz. modulated at 16 Hz of amplitude 10 V peak to peak) on sciatic neurectomy induced osteoporosis in rat femur and tibia resulted in statistically significant increase in bone mineral density, and deceleration in bone resorption process and consequently further osteoporosis in rat bone. These results suggest that such an effective window of pulsed radio frequency fields may be used therapeutically for the treatment of osteoporosis.  相似文献   

7.
Hokenson MJ  Cope GA  Lewis ER  Oberg KA  Fink AL 《Biochemistry》2000,39(21):6538-6545
Class A beta-lactamases hydrolyze penicillins and other beta-lactams via an acyl-enzyme catalytic mechanism. Ser70 is the active site nucleophile. By constructing the S70A mutant, which is unable to form the acyl-enzyme intermediate, it was possible to make stable ES complexes with various substrates. The stability of such Michaelis complexes permitted acquisition of their infrared spectra. Comparison of the beta-lactam carbonyl stretch frequency (nu(CO)) in the free and enzyme-bound substrate revealed an average decrease of 13 cm(-)(1), indicating substantial strain/distortion of the lactam carbonyl when bound in the ES complex. Interestingly, regardless of the frequency of the C=O stretch in the free substrate, when complexed to Bacillus licheniformis beta-lactamase, the frequency was always 1755 +/- 2 cm(-)(1). This suggests the active site environment induces a similar conformation of the beta-lactam in all substrates when bound to the enzyme. Using deuterium substitution, it was shown that the "oxyanion hole", which involves hydrogen bonding to two backbone amides, is the major source of the enzyme-induced strain/distortion. The very weak catalytic activity of the S70A beta-lactamase suggests enzyme-facilitated hydrolysis due to substrate distortion on binding to the enzyme. Thus the binding of the substrate in the active site induces substantial strain and distortion that contribute significantly to the overall rate enhancement in beta-lactamase catalysis.  相似文献   

8.
The present study was designed to compare the morphological changes occurring in pancreatic tissue fragments transplanted into the anterior eye chamber (AEC) and the subcutaneous (SC) regions of the rat. Pancreatic tissue segments were removed from the tail end of the pancreas of neonatal rats and transplanted into the AEC and SC region of the neck of homologous rats. Five weeks after transplantation, the grafts were removed and processed for light microscopy, immunohistochemistry and radioimmunoassay. In both pancreatic tissue grafts, the acinar cells degenerated completely after transplantation. In contrast to this, insulin-, glucagon-, somatostatin- and pancreatic polypeptide-positive cells and pancreatic ducts survived equally well in both the AEC and SC grafts. The pattern and percentage distribution of insulin-, glucagon-, somatostatin- and PP-producing cells in the AEC and SC grafts was similar to that observed in normal pancreas. However, the percentage distribution of glucagon- and PP-containing cells was significantly (p < 0.03) lower in SC grafts when compared to normal. Radioimmunoassay showed that the AEC and SC pancreatic tissue grafts contained large quantities of insulin and glucagon. However, the insulin content of AEC was slightly but not significantly higher than that of SC grafts. The protein content of pancreatic tissue grafts in these transplantation sites was still significantly (p < 0.05) lower compared to normal. Lymphatic infiltration was also more conspicuous in SC grafts compared to AEC grafts. This infiltration by lymphatic cells was confined only to the endocrine portion of the graft. In conclusion, pancreatic tissue grafts survived in both the AEC and SC regions of rats but the AEC appears to be more conducive to graft survival than the SC region.  相似文献   

9.
目的:绝经后骨质疏松是好发于中老年女性人群中的骨代谢疾病,去卵巢骨质疏松大鼠模型是国内外通用的模拟绝经后骨质疏松发生的经典动物模型,本研究通过观察去卵巢骨质疏松大鼠股骨骨微结构的动态变化,为骨质疏松大鼠模型的临床应用提供理论参考依据。方法:将90只3月龄雌性SD大鼠按体重分层后随机分为基础组(10只)、假手术组(40只)和去卵巢组(40只)。分别在手术前(基础组)和后的3、6、12、24周,腹主动脉取血处死基础组以及假手术组和去卵巢组大鼠,每组各8-10只。每组中随机取6只大鼠,对其左股骨行micro-CT扫描及三维结构重建。选择股骨远端距生长板远端1 mm处,2.0 mm×3.5 mm,厚0.9 mm的骨组织为感兴趣区域,对感兴趣区域进行骨形态计量学分析。结果:与0周组比较,从去卵巢3周开始一直持续到24周,去卵巢组大鼠股骨vBMD、BV/TV和Tb.N显著降低,Tb.Sp和SMI显著升高,而Tb.Th无显著变化;与0周组比较,从假手术后3周开始一直到24周,假手术组所有检测指标均无显著变化。与同周龄假手术组比较,从去卵巢3周开始一直持续到24周,去卵巢组大鼠股骨Tb.N、BV/TV和vBMD显著降低,Tb.Sp显著升高,而Tb.Th没有显著变化。从去卵巢6周开始一直到24周,去卵巢组大鼠SMI显著增加。结论:3月龄大鼠股骨远端的骨微结构在去卵巢3周时就出现显著变化。提示,采用3月龄大鼠进行抗骨质疏松药物筛选时,去卵巢3周后就可以进行药物处理。  相似文献   

10.
The dynamics of the ferric CN complexes of the heme proteins Myoglobin and Hemoglobin I from the clam Lucina pectinata upon Soret band excitation is monitored using infrared and broad band visible pump-probe spectroscopy. The transient response in the UV-vis spectral region does not depend on the heme pocket environment and is very similar to that known for ferrous proteins. The main feature is an instantaneous, broad, short-lived absorption signal that develops into a narrower red-shifted Soret band. Significant transient absorption is also observed in the 360-390 nm range. At all probe wavelengths the signal decays to zero with a longest time constant of 3.6 ps. The infrared data on MbCN reveal a bleaching of the C triple bond N stretch vibration of the heme-bound ligand, and the formation of a five-times weaker transient absorption band, 28 cm(-1) lower in energy, within the time resolution of the experiment. The MbC triple bond N stretch vibration provides a direct measure for the return of population to the ligated electronic (and vibrational) ground state with a 3-4 ps time constant. In addition, the CN-stretch frequency is sensitive to the excitation of low frequency heme modes, and yields independent information about vibrational cooling, which occurs on the same timescale.  相似文献   

11.
The role played by nonspecialized connective tissues in chronic non-specific low back pain is not well understood. In a recent ultrasound study, human subjects with chronic low back pain had altered connective tissue structure compared to human subjects without low back pain, suggesting the presence of inflammation and/or fibrosis in the low back pain subjects. Mechanical input in the form of static tissue stretch has been shown in vitro and in vivo to have anti-inflammatory and anti-fibrotic effects. To better understand the pathophysiology of lumbar nonspecialized connective tissue as well as potential mechanisms underlying therapeutic effects of tissue stretch, we developed a carrageenan-induced inflammation model in the low back of a rodent. Induction of inflammation in the lumbar connective tissues resulted in altered gait, increased mechanical sensitivity of the tissues of the low back, and local macrophage infiltration. Mechanical input was then applied to this model as in vivo tissue stretch for 10 minutes twice a day for 12 days. In vivo tissue stretch mitigated the inflammation-induced changes leading to restored stride length and intrastep distance, decreased mechanical sensitivity of the back and reduced macrophage expression in the nonspecialized connective tissues of the low back. This study highlights the need for further investigation into the contribution of connective tissue to low back pain and the need for a better understanding of how interventions involving mechanical stretch could provide maximal therapeutic benefit. This tissue stretch research is relevant to body-based treatments such as yoga or massage, and to some stretch techniques used with physical therapy.  相似文献   

12.
Bone mineral density (BMD) of the whole body and hind limb of young adult rats, with and without a sham-operated stifle joint was studied, using dual energy x-ray absorptiometry (DEXA) at three time points. Data from the whole body scan were used for analyses of BMD, bone mineral content (BMC), fat, lean, body weight (BW), percentage of BMC (%BMC), percentage of fat (%fat), and percentage of lean (%lean), none of which were significantly different between the groups at any time point. Significant (P < 0.05) differences in BMD, BMC, %BMC, BW, fat, %fat, and %lean were apparent at the second and third scans, compared with the initial scan, within both groups. Changes in whole body BMD, BMC, and %BMC as well as BW were highly correlated with time in both groups. In the hind limb scans, regions of interest (ROIs) were created to obtain values of BMD and BMC from the whole femur, whole tibia including the fibula, distal portion of the femur, and proximal portion of the tibia. Significant differences were not found between the groups for any ROIs. However, significant BMD and BMC increases were evident in all ROIs at the second and third scans, compared with the initial scan. Similar to those in the whole body scan, BMD and BMC obtained from ROIs were highly correlated with time. The positioning technique for the whole body and appendicular scans was analyzed by calculating percentage of the coefficient of variation (%CV) at the beginning of the study. The %CV was low and acceptable in ROIs for the hind limb and for all parameters of the whole body scan, except fat. The results suggest that in vivo DEXA scanning of the rat whole body and appendicular skeleton is highly reproducible and useful to study the whole skeleton, as well as a region of a long bone of the rat. Values for the sham-operated rats were not significantly different from those for the untreated controls, which suggests that soft tissue damage around the stifle joint did not alter BMD in the subchondral bone of the distal portion of the femur and proximal portion of the tibia.  相似文献   

13.
Fourier transform infrared (FTIR) spectroscopy was used to examine the effect of oxysterol insertion into normal and sickle RBC membranes and the total lipid extracts of the membranes. Examination of the FTIR C-H stretch and fingerprint regions reveal that the insertion of 7 alpha- and 7 beta-hydroxycholesterol has the greatest effect on the fluidity of RBC membranes and lipid extracts. The results confirm the observation that sterol molecules are oriented in the membrane so that the 7 position is located in the phospholipid head group region at the lipid/water interface. The substitution of a keto for a hydroxy group at the number seven position decreases the effect of the sterol on membrane packing.  相似文献   

14.
We have investigated the phase characteristics of 1,2-bis(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine (DC23PC), a phosphatidylcholine with diacetylenic groups in the acyl chains, and its saturated analog 1,2-ditricosanoyl-sn-glycero-3-phosphocholine (DTPC), using Fourier-transform infrared spectroscopy (FTIR). Previous studies on the phase behavior of DC23PC in H2O have shown that DC23PC exhibits: (1) formation of cylindrical structures ('tubules') by cooling fluid phase multilamellar vesicles (MLVs) through Tm (43 degrees C), and 2) metastability of small unilamellar vesicles (SUVs) in the liquid-crystalline state some 40 degrees C below Tm, with subsequent formation of a gel phase comprised of multilamellar sheets at 2 degrees C. The sheets form tubules when heated and cooled through Tm. FTIR results presented here indicate that as metastable SUVs are cooled toward the transition to bilayer sheets, spectroscopic changes occur before the calorimetric transition as measured by a reduction in the CH2 symmetric stretch frequency and bandwidth. In spite of the vastly different morphologies, the sheet gel phase formed from SUVs is spectroscopically similar to the tubule gel phase. The C-H stretch region of DC23PC gel phase shows bands at 2937 and 2810 cm-1 not observed in the saturated analog of DC23PC, which may be related to perturbations in the acyl chains introduced by the diacetylenic moiety. The narrow CH2 scissoring mode at 1470 cm-1 and the prominent CH2 wagging progression indicate that DC23PC gel phase was highly ordered acyl chains with extended regions of all-trans methylene segments. In addition, the 13 cm-1 reduction in the C = O stretch frequency (1733-1720 cm-1) during the induction of DC23PC gel phase indicates that the interfacial region is dehydrated and rigid in the gel phase.  相似文献   

15.
Fourier transform infrared (FT-IR) microspectroscopy is a powerful technique that can be used to collect infrared spectra from microscopic regions of tissue sections. The infrared spectra are evaluated to chemically characterize the absorbing molecules. This technique can be applied to normal or diseased tissues. In the latter case, FT-IR microspectroscopy can reveal chemical changes that are associated with discrete regions of lesion sites, which can provide insights into the chemical mechanisms of disease processes. In the present study, FT-IR microspectroscopy was used to analyze sections of retina from normal (pigmented) and albino rats. The outer segments of retinas from pigmented animals were found to have unusually strong absorption values for C&z.dbnd6;C-H unsaturation and carbonyl functional groups. Docosahexaenoic acid (DHA), a major constituent of lipids in the outer segments, also had particularly high absorption values for these functional groups, which suggests that it is responsible for those enhanced absorption values. Absorbance values for the unsaturation and carbonyl functional groups were substantially reduced in the outer segments of retinas from albino animals. This finding, together with data from other studies on light-induced oxidative events in the retina, indicates a loss of DHA by a light-induced mechanism in albino animals. The outer nuclear layer had strong absorbance values for H-C-OH and P&z. dbnd6;O functional groups, which is likely due to the sugar phosphate backbone of DNA. The outer and inner plexiform layers were found to contain greater concentrations of CH(2) and C&z.dbnd6;O functional groups than the outer and inner nuclear layers, which is due to the high concentration of synaptic connections in the former layers. In summary, FT-IR microspectroscopy revealed a unique chemical profile in the outer segments compared to other retinal layers, and this profile was altered in albino animals.  相似文献   

16.
This investigation of microstructure in the human proximal femur probes the relationship between the parameters of the FRAX index of fracture risk and the parameters of bone microstructure. The specificity of fracture sites at the proximal femur raises the question of whether trabecular parameters are site-specific during post-menopause, before occurrence of fragility fracture. The donated proximal femurs of sixteen post-menopausal women in the sixth and seventh decades of life, free of metabolic pathologies and therapeutic interventions that could have altered the bone tissue, constituted the material of the study. We assessed bone mineral density of the proximal femurs by dual energy X-ray absorptiometry and then sectioned the femurs through the center of the femoral head and along the femoral neck axis. For each proximal femur, morphometry of trabeculae was conducted on the plane of the section divided into conventional regions and sub-regions consistent with the previously identified trabecular families that provide regions of relatively homogeneous microstructure. Mean trabecular width and percent bone area were calculated at such sites. Our findings indicate that each of mean trabecular width and percent bone area vary within each proximal femur independently from each other, with dependence on site. Both trabecular parameters show significant differences between pairs of sites. We speculate that a high FRAX index at the hip corresponds to a reduced percent bone area among sites that gives a more homogeneous and less site-specific quality to the proximal femur. This phenomenon may render the local tissue less able to carry out the expected mechanical function.  相似文献   

17.
Multipotent stem cells are considered as a key material in regenerative medicine, and the understanding of the heterogeneity in the differentiation potentials of bone marrow-derived cells is important in the successful regenerative tissue repair. Therefore, the present study has been performed to investigate how the differentiation of post-harvest, native bone marrow-derived cells is regulated by cyclic stretch in vitro. Bone marrow-derived cells were obtained from mouse femur of both hind limbs and categorized into the following five categories: amebocytes, round cells, spindle cells, stellate cells and others. The cells were seeded on a silicone-made stretch chamber, and subjected to cyclic stretch with an amplitude of 10% at a frequency of 1 Hz for 7 days for cell shape analysis and for 3 days for the analysis of the expression of marker proteins of osteogenic (osteocalcin), vascular smooth muscle (α-smooth muscle actin and smooth muscle myosin heavy chain) and neurogenic (neurofilament) differentiation. When disregarding the differences in the cell shapes, there was an overall trend that the application of 10% cyclic stretch inhibited osteogenic and neurogenic differentiation, but enhanced smooth muscle differentiation. Close examinations revealed that round cells were influenced the most by cyclic stretch (significant up- or down-regulation in all the four marker protein expressions) while amebocytes and spindle cells were only influenced by cyclic stretch for vascular smooth muscle and/or neurogenic differentiation. As far as the authors know, this is the first study reporting the shape-related differences in the fate decision criteria for mechanical strain in bone marrow-derived cells.  相似文献   

18.
Nucleotide variation in populations originating from the recent range expansion of a species should reflect their adaptation to new habitats as well as their demographic history. A survey of nucleotide variation at 109 noncoding X-chromosome fragments in a European population of Drosophila melanogaster allowed identifying some candidates to have been recently affected by positive selection. Adaptive changes leave a spatial differential footprint that can be used to discriminate among candidates by extending their study to neighboring regions. Here, we surveyed variation at an approximately 190-kb region spanning a locus exhibiting a significantly skewed frequency spectrum. A stretch of approximately 12 kb with reduced variation was detected within a continuously sequenced region that included the focal fragment. Moreover, the regions flanking this stretch exhibited an excess of high-frequency derived variants. Application of maximum likelihood ratio and goodness-of-fit tests suggested that the pattern of variation detected at the studied region (at cytological bands 17C-17D) might have been shaped by a recent selective change, most probably at or around the phantom gene that encodes CYP306A1, a cytochrome P450 enzyme in the ecdysteroidogenic pathway.  相似文献   

19.
Femoroplasty is a potential preventive treatment for osteoporotic hip fractures. It involves augmenting mechanical properties of the femur by injecting Polymethylmethacrylate (PMMA) bone cement. To reduce the risks involved and maximize the outcome, however, the procedure needs to be carefully planned and executed. An important part of the planning system is predicting infiltration of cement into the porous medium of cancellous bone. We used the method of Smoothed Particle Hydrodynamics (SPH) to model the flow of PMMA inside porous media. We modified the standard formulation of SPH to incorporate the extreme viscosities associated with bone cement. Darcy creeping flow of fluids through isotropic porous media was simulated and the results were compared with those reported in the literature. Further validation involved injecting PMMA cement inside porous foam blocks — osteoporotic cancellous bone surrogates — and simulating the injections using our proposed SPH model. Millimeter accuracy was obtained in comparing the simulated and actual cement shapes. Also, strong correlations were found between the simulated and the experimental data of spreading distance (R2 = 0.86) and normalized pressure (R2 = 0.90). Results suggest that the proposed model is suitable for use in an osteoporotic femoral augmentation planning framework.  相似文献   

20.
Fourier transform infrared spectroscopy was used to characterize the interaction of the cationic lipids 1,2-dioleoyl-3-trimethylammonium-propane and dioctadecyldimethylammonium bromide with plasmid DNA. The effect of incorporating the neutral colipids cholesterol and dioleoylphosphatidylethanolamine on this interaction was also examined. Additionally, dynamic and phase analysis light scattering were used to monitor the size and zeta potential of the resulting complexes under conditions similar to the Fourier transform infrared measurements. Results suggest that upon interaction of cationic lipids with DNA, the DNA remains in the B form. Distinct changes in the frequency of several infrared bands arising from the DNA bases, however, suggest perturbation of their hydration upon interaction with cationic lipids. A direct interaction of the lipid ammonium headgroup with and dehydration of the DNA phosphate is observed when DNA is complexed with these lipids. Changes in the apolar regions of the lipid bilayer are minimal, whereas the interfacial regions of the membrane show changes in hydration or molecular packing. Incorporation of helper lipids into the cationic membranes results in increased conformational disorder of the apolar region and further dehydration of the interfacial region. Changes in the hydration of the DNA bases were also observed as the molar ratio of helper lipid in the membranes was increased.  相似文献   

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