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Structural changes in aging bone: osteopenia in the proximal femurs of female mice.

A computerized image analysis system was used to quantitate age-related changes in the structure of the proximal femur in CW-1 female mice, ranging from 3 to 32 months of age. Morphological findings revealed a progressive thinning of bone trabeculae within the femoral head, accompanied by the development of marrow cavities in the cortical bone of the femoral neck and in the subchondral bone. As a result, the compact bone in senescent mice acquired an appearance similar to trabecular bone. Quantitative image analysis revealed a similarity in the pattern of changes in the three types of bone: cortical, trabecular, and subchondral. Bone density increased from 3 to 12 months of age and subsequently declined. A similar pattern was noted for the changes in the thickness of the cortical and the subchondral bone. Regression analysis revealed that the changes with age fitted a second-order model; thus it was possible to predict the age of maximal values for each parameter. Hence, the age of maximal bone density for cortical, trabecular, and subchondral bone was 12.3, 14.8, and 18.0 months, respectively. The rate of bone loss after 12 months was most prominent for trabecular bone (1.47% per month), so that by 32 months of age its overall mass had declined by 57% in comparison to peak values seen at 12 months of age (p less than 0.001). The density of the subchondral and cortical bones decreased at a slower rate (0.6% to 0.8% per month) and at the age of 32 months their values had decreased by 12% to 18% in comparison to those at 12 months (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Aging

Phenytoin neuropathy: structural changes in the sural nerve.

Phenytoin has been implicated as a causative agent in peripheral neuropathy, although structural changes in nerve have not been characterized. A 47-year-old man was seen with clinical and electrophysiological signs of peripheral neuropathy after 30 years of phenytoin administration. Despite a modest dose of phenytoin (300 mg/day) blood levels were 31 to 38 micrograms/ml. A sural nerve biopsy showed a loss of large myelinated nerve fibers and a nonrandom clustered distribution of segmental demyelination and remyelination. The latter findings were accompanied by axonal shrinkage. Sixteen months after phenytoin was stopped, the patient's clinical and electrophysiological findings reflected improvement. These data indicate that long-term phenytoin administration can cause a reversible neuropathy characterized by axonal shrinkage and secondary demyelination.

Atrophy

Nuclease-induced DNA structural changes assessed by flow cytometry with the intercalating dye propidium iodide.

A flow cytometric analysis of DNA structural changes induced by cleavage with nucleases was performed on isolated HeLa nuclei by assessing changes in stainability with the DNA-specific fluorochrome propidium iodide (PI). After mild digestion with DNAse I, micrococcal nuclease, or with the single-strand-specific S1 and Neurospora crassa nucleases, fluorescence intensity of nuclei stained with PI increased by about 15-30% above the value of undigested control samples. No significant modifications were observed with the restriction enzymes Eco RI, Alu I, and Not I. The DNAse I-induced increase in fluorescence intensity was also observed with the non-intercalating dye Hoechst 33258, but not with mithramycin. Nuclease-induced fluorescence intensity changes as determined with PI were found to be dependent on the dye concentration. A constant increase (about 20%) was measured at dye/DNA-P ratios greater than 0.11. Below this value (2 micrograms/ml PI), the fluorescence intensity of digested samples was 15-30% lower than that of undigested controls. This behaviour towards intercalating dyes is similar to that of the relaxed (nicked) vs. the supercoiled (intact) form of circular DNA. These results suggest that conformation- but not sequence-specific nucleases induce a relaxation of DNA supercoils.

Coloring Agents

Acid-induced structural changes of a mouse IgG2a monoclonal antibody (MN12) studied by transient electric birefringence measurement.

Acid-induced structural changes of a mouse IgG2a monoclonal antibody (MN12) as indicated by Jiskoot et al. (Eur. J. Biochem. 201,223-232 (1991)) were studied by measuring the transient electric birefringence of MN12 in aqueous solution and in glycerol-water mixtures at different pH conditions. A multi-exponential analysis program, DISCRETE (Provencher,S.W., Biophys.J.16,27-41 (1976)), and a constrained inverse Laplace transform program, CONTIN (Provencher, S.W., Comp. Phys. Comm. 27, 213-227 (1982)) have been used to determine the number of exponentials needed to represent the data and their decay times. Measurement of the time-resolved electric field induced birefringence makes it possible to study rotational processes on a timescale from several tens of nanoseconds to microseconds. This enabled us to monitor the segmental flexibility and the rotational motion of single antibody molecules as well as the occurrence of aggregates. The results show an increase in hydrodynamic dimensions of MN12 upon lowering the pH from 6.6 to 2.7. Additionally, the original segmental flexibility, which could be monitored for the samples in glycerol-water mixtures, is altered at low pH. The results have been interpreted as swelling of MN12 followed by dimerization.

Animals

Fine-structural changes in the nucleus of primordial oocytes in immature hamsters.

This study reports observations on fine-structural changes in the nucleus of the oocyte in hamsters during the establishment of the primordial (unilaminar) follicle at 7 to 12 days after birth. Following pachytene, the nuclei at early diplotene contain simple chromosomal cores surrounding by a sheath of fibrillo-granular material symmetrically disposed in the form of lateral projections and loops. From 9 days onwards increasing numbers of the oocytes show nuclei lacking such chromosomal threads: instead, they contain randomly disposed, condensed fibrillar clumps with associated dense knots of material, and prominent aggregations of 25--30-nm granules. This second nuclear form is assumed by all oocytes examined at 11 and 12 days and appears to be the definitive diplotene phase. These changes in the appearance of the nuclear chromatin are discussed in the light of those found in oocytes of other species, and it is concluded that the hamster diplotene phase consists of a transitory 'compace' or uniform lampbrush stage, followed by a definitive dictyate condition like that in the rat, but with markedly differential condensation or extreme diffusion of lateral loops. This conclusion is examined in relation to findings which report a marked increase in the sensitivity of hamster oocytes to X-radiation at the time that the observed changes in nuclear configuration are taking place. The combined observations support the hypothesis that the dictyate stage in rodent oocytes represents a modified diplotene phase in which the high degree of spatial diffusion of components of chromosomes of the lampbrush type results in a reduced capacity for repair following exposure to ionizing radiations.

Animals

[Structural changes in the liposomes and the lamellar packing of liposomal lipids exposed to glycerin].

Glycerol-induced structural changes in multilamellar liposomes (ML) were studied by light and thin-section electron microscopy. It is shown that high concentrations of glycerol induce non-bilayer rearrangements of liposomal lipids in ML. The data obtained are discussed regarding the possible mechanisms of cryoprotector effects on natural (biological) and artificial (liposomal) membranes.

Animals

[Roentgen diagnosis of structural changes in the pubic area in children and adolescents. Personal case reports, differential diagnosis and literature review].

Structural changes in the pubic bone often are difficult to interpret. The radiologic appearance is very different. The differentiation between inflammatory and malignant processes is hardly to distinguish. We are reporting ten patients with osseous changes in the os pubic (4 Ewing's sarcoma, 1 osteosarcoma, 1 Morbus Hodgkin, 1 osteochondronecrosis, 2 osteochondritis, 1 subchronic osteomyelitis). Two patients with Ewing's sarcoma who were initially treated for osteomyelitis are described in detail. Differential diagnosis and previous literature are reviewed.

Adolescent

Chromatin structure changes suggest a compensatory response to c-myc gene amplification in malignant fibrous histiocytoma.

Changes in chromatin structure as determined from DNAse I hypersensitive site analysis are associated with c-myc amplification and increased transcript/protein levels in malignant fibrous histiocytoma (MFH) cell lines. A DNAse I hypersensitive site near the PO promoter region was observed in one MFH cell line (UR HCL 1), and in normal fibroblasts (HFF), but not in an MFH cell line with an amplified c-myc gene (P3C). A DNAse I hypersensitive site exclusive to P3C amplified c-myc was identified slightly 3' of exon one. No alterations in c-myc DNAse I hypersensitive site patterns were observed in HFF fibroblasts following serum release, when peak levels of c-myc transcript were induced. DNAse I hypersensitive site patterns associated with gene amplification may reflect a compensatory response by P3C cells to an abundance of c-myc transcript. Furthermore, elevated levels of protein in P3C cells provide additional evidence that amplified c-myc is an oncogene in MFHs.

Chromatin

Structural changes in membranes of large unilamellar vesicles after binding of sodium cholate.

The interaction of the bile salt cholate with unilamellar vesicles was studied. At low cholate content, equilibrium binding measurements with egg yolk lecithin membranes suggest that cholate binds to the outer vesicle leaflet. At increasing concentrations, further bile salt binding to the membrane is hampered. Before the onset of membrane solubilization, diphenylhexatriene fluorescence anisotropy decreases to a shallow minimum. It then increases to the initial value in the cholate concentration range of membrane solubilization. At still higher cholate concentrations, a drop in fluorescence anisotropy indicates the transformation of mixed disk micelles into spherical micelles. Perturbation of the vesicle membranes at molar ratios of bound cholate/lecithin exceeding 0.15 leads to a transient release of oligosaccharides from intravesicular space. The cholate concentrations required to induce the release depend on the size of the entrapped sugars. Cholesterol stabilizes the membrane, whereas, in spite of enhanced membrane order, sphingomyelin destabilizes the membrane against cholate. Freeze-fracture electron microscopy and phosphorus-31 nuclear magnetic resonance (31P NMR) also reflect a change in membrane structure at maximal cholate binding to the vesicles. In 31P NMR spectra, superimposed on the anisotropic line typically found in phospholipid bilayers, an isotropic peak was found. This signal is most probably due to the formation of smaller vesicles after addition of cholate. The results were discussed with respect to bile salt/membrane interactions in the liver cell. It is concluded that vesicular bile salt transport in the cytoplasm is unlikely and that cholate binding is restricted to the outer leaflet of the canalicular part of the plasma membrane.

1,2-Dipalmitoylphosphatidylcholine

Cyclic AMP-dependent protein kinase I: cyclic nucleotide binding, structural changes, and release of the catalytic subunits.

Type I cyclic AMP (cAMP)-dependent protein kinase is composed of a dimeric regulatory subunit (R(2)) and two catalytic subunits (C subunits). The R(2) dimer binds four cAMP molecules to release the two C subunits. To characterize the cAMP binding sites and elucidate their role in the release of the C subunits, the R(2) dimer has been studied by equilibrium methods. The cAMP titration of R(2) was monitored by endogenous tryptophan fluorescence, and the results suggest one class of binding sites. The titration plot is monotonic for saturation of four sites per R(2). A similar titration monitored by near-UV circular dichroic changes exhibited profound changes in the region of the (1)L(b) tyrosine and (1)L(a) and (1)L(b) tryptophan transitions; a plot of these data also showed a linear monotonic response. Thus, the fluorescence and circular dichroic changes show that cAMP binding to R(2) induces a conformational or structural change. The one apparent class of binding sites implies that all binding sites are characterized by similar K(d) values or by K(d) values much less than the receptor concentration. The reactivity of the cysteine sulfhydryl groups with 5,5'-dithiobis(2-nitrobenzoic acid) showed that saturation with cAMP indirectly protects one sulfhydryl group per R monomer. Analysis of cAMP activation of the holoenzyme, detected by phosphotransferase assays, showed that saturation of both cAMP binding sites per R monomer is necessary to effect the release of the C subunit. By using a fluorescent analog of cAMP, 1,N(6)-etheno-cyclic AMP (epsilon cAMP), the (epsilon cAMP)(4).R(2) complex was titrated with C subunit, causing the release of epsilon cAMP. The titration showed that the release of epsilon cAMP was a positive cooperative process; its Hill plot had a slope of 2.6 and the K(a1) and K(an) values obtained by extrapolation were 2.1 x 10(7) M(-1) and 5.0 x 10(8) M(-1), respectively. The calculated DeltaDeltaG for first and last site coupling was 1.9 kcal/mol (1 cal = 4.18 J) of holoenzyme.

Animals

Structural changes in the membrane of vero cells infected with a paramyxovirus.

Vero cells productively infected with the Halle strain of measles virus have been studied by means of surface replication, freeze-fracturing, and surface labeling with horseradish peroxidase-measles antibody conjugate in order to examine changes in the structure of the cell membrane during viral maturation. Early in infection, the surfaces of infected cells are embossed by scattered groups of twisted strands, and diffuse patches of label for viral antigens cover regions marked by these strands. At later stages, when numerous nucleocapsids become aligned under the plasmalemmal strands, the strands increase in number and width and become more convoluted. At this stage, label for viral antigens on the surface of the cell membrane is organized into stripes lying on the crests of strands. Finally, regions of the membrane displaying twisted strands protrude to form ridges or bulges, and the freeze-fractured membrane surrounding these protrusions is characterized by an abundance of particles small than those found on the rest of the cell membrane. The fractured membranes of viral buds are continuous sheets of these small particles, and the spacing between both nucleocapsids and stripes of surface antigen in buds is less than in the surrounding cell membrane. Detached virus is covered with a continuous layer of viral antigen, has unusually large but no small particles on its membrane surfaces exposed by freeze-fracturing, and no longer has nucleocapsids aligned under its surface. Thus, surface antigens, membrane particles, and nucleocapsids attached to the cell membrane are mobile within the plane of the membrane during viral maturation. All three move simutaneously in preparation for viral budding.

Antigens, Viral

The sheep ovary: regulation of steroidogenic, haemodynamic and structural changes in the largest follicle and adjacent tissue before ovulation.

Important changes occur in the follicle that is destined to ovulate and also in the surrounding ovarian tissue of sheep during the 72 hr preceding ovulation. It is our purpose to describe the morphological and functional changes that take place in large follicles during the preovulatory period and to relate these to the local ovarian environment and general endocrine status prevailing at this time. A consideration of changes in the structure, blood supply and function of the ovaries in vivo, is followed by a discussion of the functional capacity of the individual ovarian components when isolated and studied in vitro. Against this background, the nature and sequence of events that occur in the preovulatory follicle and the mechanisms which regulate them will be discussed.

Adenylyl Cyclases

[Structural changes in the jaws. Differential diagnosis].

In this work the Authors have considered the causes of changes in the structure of maxillary bones. They have also discussed the available diagnostic means useful to make a correct diagnosis and the importance of a right differential diagnosis of the causes in order to have a better treatment.

Diagnosis, Differential

Histone phosphorylation in native chromatin induces local structural changes as probed by electric birefringence.

In order to understand how the phosphorylation of histones affects the chromatin structure, we used electron microscopy, sedimentation velocity, circular dichroism and electric birefringence to monitor the salt-induced filament reversible solenoid transition of phosphorylated and native chromatin. Phosphorylation in vitro of chicken erythrocyte chromatin by cyclic-AMP-dependent protein kinase from porcine heart led to the modification of the histones H3 and H5 only, which were modified at a level of one phosphate and about three phosphate groups per molecule, respectively. In contrast to circular dichroism and sedimentation studies, which tend to suggest that phosphorylation of H3 and H5 does not affect chromatin structure, electron microscopy reveals that phosphorylation causes a relaxation of structure at low ionic strength. Electric birefringence and relaxation time measurements clearly prove that local structural changes are induced in chromatin: we observe a decrease of the steady-state birefringence with the appearance of a negative contribution in the signal and a marked increase of the flexibility of fibres. The component with the negative birefringence presents very short relaxation times, like those exhibited by small DNA fragments or individual nucleosomes. Two possibilities are then suggested. First, the conformational change is consistent with what would be expected from the presence of DNA segments loosely associated with the core histone H3. That the length of such segments could correspond to about one to two base-pairs per nucleosome strongly suggests that phosphorylation induces changes affecting some specific H3-DNA interactions only. This result could corroborate previous observations indicating that the N-terminal region of H3, where the site of phosphorylation is located, plays a decisive role in maintaining the superstructure of chromatin. Second, phosphorylation could introduce hinge points between each nucleosome. In this case, the negative birefringence results from partial orientation of the swinging nucleosomes. A possible mode of action of phosphorylation might be to weaken structural restraints imposed by histone H3, thus facilitating further condensation of chromatin.

Animals

Does feeding of hexachlorobenzene promote structural changes in rat-liver porphyrinogen carboxy-lyase?

This study aimed to determine whether drug treatment produced structural changes in porphyrinogen carboxy-lyase enzymatic protein, leading to altered properties. Rat-liver enzyme was obtained from normal animals and from those with hexachlorobenzene (HCB)-induced porphyria, and several of its properties were comparatively studied. The enzymes from both sources were purified 110-fold. They were similar in subcellular distribution, ammonium sulfate fractionation, calcium phosphate gel adsorption, storage stability, requirements of incubation media, effects of salts and photo-oxidizing agents. The enzymes differed with respect to effect of incubation temperature, pH, aerobiosis, chelating agents, dithiothreitol, methylene blue, heat stability and chromatographic behaviour on DEAE-cellulose and Sephadex G-100. These differences would appear to indicate structural differences between the two types of enzymatic protein. Since the porphyrinogenic action of HCB can be mediated through a metabolite, structural differences could arise through the binding of an inhibitory metabolite to the whole enzyme or through modifications to the protein during its synthesis.

Animals

Structural changes in the outer retinal layers following blunt mechanical non-perforating trauma to the globe: an experimental study.

Structural changes in the external retinal layers of pigs' eyes that had previously been subjected to blunt non-perforating mechanical injuries have been described. Within minutes of trauma fragmentation of the photoreceptor outer segments and damage to the retinal pigment epithelium was recorded in areas of retina adjacent to the missile impact site on the sclera. One week after injury the disrupted photoreceptor outer segments disappeared, and the inner segments came to lie adjacent to the retinal pigment epithelium. Evidence of regeneration of photoreceptor outer segments was noted 2 weeks later.

Animals