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Synapse loss and axon retraction in response to local muscle degeneration.

During metamorphosis in the moth, Manduca sexta, the abdominal body-wall muscle DEO1 is remodeled to form the adult muscle DE5. As the larval muscle degenerates, its motoneuron loses its end plates and retracts axon branches from the degenerating muscle. Muscle degeneration is under the control of the insect hormones, the ecdysteroids. Topical application of an ecdysteroid mimic resulted in animals that produced a localized patch of pupal cuticle. Muscle fibers underlying the patch showed a gradient of degeneration. The motoneuron showed end-plate loss and axon retraction from degenerating regions of a given fiber but maintained its fine terminal branches and end plates on intact regions. The results suggest that local steroid treatments that result in local muscle degeneration bring about a loss of synaptic contacts from regions of muscle degeneration.

Animals↗

Heritabilities of radiologic osteoarthritis in peripheral joints and of disc degeneration of the spine.

OBJECTIVE: To estimate the genetic influence on the occurrence of radiologic osteoarthritis (ROA) in the knees, hips, and hands and disc degeneration of the spine in the general population. METHODS: A random sample of 1,583 individuals was drawn to estimate the prevalence of ROA and disc degeneration in the general population. Of 118 probands with multiple affected joint sites who were derived from this sample, we were able to recruit 257 siblings. The variance of ROA and disc degeneration within sibling pairs was compared with the variance between sibling pairs. Heritability estimates for ROA in the knees, hips, and hands and for disc degeneration of the spine were calculated. OA was defined according to radiologic criteria, using the Kellgren/Lawrence grading system. RESULTS: We observed that hand ROA and disc degeneration of the spine were statistically significantly more frequent in siblings than in the random sample, whereas the prevalence of knee and of hip ROA was similar and lower, respectively. Heritability estimates for hand ROA and disc degeneration were statistically significant, P = 0.56 (95% confidence interval [95% CI] 0.34-0.76) and P = 0.75 (95% CI 0.30-1.00), respectively. For knee and hip ROA, no evidence of a genetic effect in the general population was found. Finally, the heritability estimate for a score that summed the number of joints affected in the knees, hips, hands, and spine was 0.78 (95% CI 0.52-0.98). All heritability estimates were adjusted for age, sex, body mass index, and bone mineral density. CONCLUSION: The present study shows that in the general population, there is a strong genetic effect for hand ROA and disc degeneration of the spine. The findings on the total number of joints affected at multiple sites suggest genetic susceptibility to generalized OA.

Aged↗

Development and degeneration of the arterial system in the mesonephros and metanephros of chicken embryos.

BACKGROUND: The general morphology of the mesonephric and metanephric arteries in chicken embryos has already been described previously. Moreover, the general basis of glomerulogenesis has also been established. However, the degeneration of the mesonephric vascular system, and especially glomerular degeneration, have not been well established yet. Also the morphology of the metanephric angiogenic buds has not been studied yet. METHODS: Scanning electron microscopy of vascular corrosion casts and critical point dried specimens as well as light microscopy of serially sectioned material has been used in this study. Mesonephric degeneration coincides in time with metanephros growth and maturation in the developmental stages of chicken embryos chosen for this investigation (7.5, 9, 11, and 14 days of incubation). RESULTS: The arterial system of the mesonephros in embryonic chicken is similar to that of the anuran kidney, as described in the literature. The morphology of the degenerating mesonephric glomeruli shows that the glomerular capillaries are more thick, tortuous, and numerous than those in normal glomeruli. The podocytes also show degeneration. The arterial system of the metanephros grows directly from the aorta and from the mesonephric arterial system. During these stages of rapid growth, the metanephros shows angiogenic buds. These angiogenic buds can be either pointed or round blind endings. CONCLUSIONS: The distribution and topography of the mesonephric and metanephric arteries is in general accordance with the literature. The process of glomerular degeneration in the mesonephros seems similar to glomerular senescence in man but is different from that of the aged rat glomeruli. The round angiogenic buds observed in the metanephros resemble tumoral angiogenic buds in some aspects. However, both angiogenesis and the degenerative phenomena are part of the normal developmental process. Consequently, the involved mechanisms are probably under sole genetic control. The system studied here offers therefore the possibility to study vascular growth and degeneration on the same model in physiological conditions without application of vasoactive or pathological agents.

Animals↗

Nuclear degeneration in the developing lens and its regulation by TNFalpha.

DNA fragmentation in lens fibre cell nuclei undergoing programmed degeneration was identified by terminal deoxynucleotidyl transferase (TdT)-mediated biotin-dUTP nick end labelling (TUNEL). Lens epithelial cells in culture were induced to differentiate into lens fibre-like clumps of cells (lentoids) by insulin and it was shown that the TUNEL method was also an effective means of labelling degenerating nuclei in lentoid cells in lens epithelial cell cultures. Using immuno-fluorescence and confocal microscopy, it was shown that TNFalpha and TNF receptor (TNFR1, and TNFR2) immunoreactivity was present in sections of chick embryo lenses. TNFalpha immunoreactivity was associated with the lens epithelium and lens fibres. TNFR1 immunoreactivity was present in lens epithelial cells, cortical lens fibres, and lens fibre cell nuclei, while TNFR2 immunoreactivity had a similar distribution to that of TNFR1, but was not associated with nuclei. Similar patterns of TNFalpha, TNFR1, and TNFR2 immunoreactivity were observed in lens epithelial cell cultures. When added to lens epithelial cell cultures, TNFalpha, at concentrations of 50 to 100 ng ml-1, and agonistic antibodies to both TNFR1 and TNFR2 significantly (P<0.05) enhanced the number of degenerating (TUNEL-positive) nuclei. On the other hand, a neutralising antibody to TNFalpha significantly (P<0. 05) reduced the number of TUNEL-positive nuclei. These results demonstrate that TUNEL is an effective means of labelling degenerating lens fibre nuclei during lens fibre and lentoid differentiation, and suggest a potential role for TNFalpha-like factors and their receptors in the degeneration of lens fibre cell nuclei during lens differentiation. We further suggest that the nuclear degeneration of lens fibre cells is analogous to the nuclear events that occur during apoptosis, and that in lens cells the nuclear degeneration is uncoupled from the plasma membrane events of apoptosis that normally lead to cell death.

Animals↗

Triphenyl phosphite and diisopropylphosphorofluoridate produce separate and distinct axonal degeneration patterns in the central nervous system of the rat.

This study compared the neurotoxic effects of triphenyl phosphite (TPP) in the rat with those seen after exposure to diisopropylphosphorofluoridate (DFP), a compound known to produce organophosphorus-induced delayed neurotoxicity (OPIDN). Animals received either three subcutaneous injections of TPP (1184 mg/kg body wt each dose) administered at 3-day intervals or a single subcutaneous injection of DFP (4 mg/kg body wt). TPP-induced clinical signs were initially observed 2 to 18 days after the last injection and included ataxia, flaccid paresis, stereotyped alternating side-to-side movements, and circling behavior. Axonal and terminal degeneration were present in the cerebellum, vestibular nuclear complex, cochlear nuclei, and superior and inferior colliculi. The subthalamic nucleus, substantia nigra, septal region, hypothalamus, thalamus, hippocampus, and cerebral cortex also contained degenerating axons and terminals. Degeneration was particularly evident in the sensorimotor cerebral cortex, mediodorsal, ventromedial, and medial geniculate thalamic nuclei and in the magnocellular preoptic and medial mammillary nuclei of the hypothalamus. Very light degeneration was present in the gracile fasciculus and nucleus. In contrast, rats injected with DFP showed moderate degeneration in the gracile fasciculus and nucleus but did not display degeneration in any other brain region. Injections of DFP did not produce delayed onset clinical signs. The results indicate that in the rat, different central nervous system cell groups are affected by these two organophosphorus compounds and that TPP affects nuclei and tracts at all levels of the neuraxis, including those associated with higher-order processing and cognitive functions. In addition, the distinct degeneration patterns produced by these two compounds support the view that TPP-induced neurotoxicity should not be considered as a type of OPIDN, but rather as a separate category of organophosphorus-induced neurotoxicity.

Animals↗

Reducing the number of microlocations in oligonucleotide microchip matrices by the application of degenerate oligonucleotides.

The application of degenerate oligonucleotides to DNA Sequencing by Hybridisation with Oligonucleotide Matrix (SHOM) is proposed. The use of degenerate oligonucleotides is regarded as an example of pooling methods that are suitable for various laboratory procedures requiring numerous samples to be assayed. As each DNA sequence coded by four letters (A, G, C, T) may be defined by two sequences: a sequence coded by W and S (W-weak-A or T, S-strong-G or C) and a sequence coded by R and Y (R-purine-A or G, Y-pirymidine-T or C), n4n -nucleotide sequences may be defined with the help of 2xn2sequences. In the place of the originally described microchip matrix composed of all possible unambiguous octanucleotides (4(8)=65 536) attached to the equal number of 65 536 microlocations a matrix composed of 512 microlocations containing 256 2(8)-degenerate octanucleotides is proposed. The matrix contains all 256 possible octanucleotides coded by W and S variations and all 256 possible octanucleotides coded by R and Y variations. The 512 256-degenerate octanucleotides allows to retrieve the same information as 65 536 unambiguous octanucleotides. A variant of the DNA sequence reconstruction method applicable to this system is presented. The use of degenerate oligonucleotides also gives the possibility to apply matrices composed of longer oligonucleotides without increasing the number of microlocations in matrices, which would enable increasing the length of unambiguously reconstructed sequence, e.g. a matrix comprising 131 072 16-mer oligonucleotides i.e. 65 536 65 536-fold degenerate oligonucleotide coded by W and S variations and 65 536 65 536-fold degenerate oligonucleotide coded by R and Y variations could replace one matrix comprising all possible unambiguous 16-mer oligonucleotides (ca. 4.3x10(9)).

Animals↗

Lumbar disc degeneration and segmental instability: a comparison of magnetic resonance images and plain radiographs of patients with low back pain.

We analyzed disc space height, angular displacement, tilting movement, and horizontal displacement in 109 patients with low back pain and/or sciatica, on plain radiographs of the lumbar spine. These parameters were compared with the grade of disc degeneration as evaluated by magnetic resonance imaging with the aim of studying lumbar segmental instability. Disc space height decreased in proportion to the grade of disc degeneration. Angular displacement was significantly less with severe disc degeneration, accompanied by a tendency to stabilization of the motion segment. Tilting movement and horizontal displacement did not correlate with the grade of disc degeneration. Lumbar segmental instability was recognized at all levels even in individuals who appeared to be normal or to have mild disc degeneration. The incidence of lumbar segmental instability at the L3-4 level was significantly higher in patients with normal discs or mild disc degeneration. At the L4-5 and L5-S levels it did not differ between different grades of disc degeneration.

Adolescent↗

Pheochromocytomas: tendency to degenerate and cause paroxysmal hypertension.

Eighty-four patients with pheochromocytoma underwent surgery during the 13-year period from 1981 to 1994. These patients were divided into three groups according to the degree of parenchymal degeneration, which was estimated by preoperative dynamic computed tomography (CT) scans and resected specimens: solid group, cystic group, and highly cystic group. The relation between the degree of parenchymal degeneration and the clinical variety, size of the tumor, predominant plasma catecholamine, and type of blood pressure disturbance was analyzed. There were 39 cases of solid pheochromocytoma, 37 cases of cystic tumor, and 8 cases of highly cystic pheochromocytoma. The maximal mean diameters for each group were as follows: 5.1 +/- 2.3, 6.8 +/- 3.0, and 7.9 +/- 3.2 cm, respectively. Of the tumors < 3 cm in diameter, 84% were solid; when the tumors were >/= 6 cm in diameter, approximately 50% were cystic and 20% highly cystic. Of the adrenal pheochromocytomas, 60% were degenerated; 79% of the extraadrenal tumors were solid. Of the epinephrine-secreting tumors, 70% were degenerated, and 55% of the norepinephrine-secreting tumors were solid. The clinical pattern of hypertension was related to the degree of degeneration: In the paroxysmal hypertension group 87% of the tumors were degenerated, and in the sustained hypertension group 69% were solid. We therefore believe that parenchymal degeneration of pheochromocytoma is highly correlated with the tumor diameter, location of the tumor, secreted catecholamine, and clinical pattern of hypertension.

Adolescent↗

The time course of elastin fiber degeneration in a rat aneurysm model.

Previous findings vary regarding the timing and cause of elastin fiber degeneration in the elastase-induced rat abdominal aortic aneurysm model. We examined the timing and cause of elastin fiber degeneration after elastase infusion using two different elastase infusion times. Twenty-four Sprague-Dawley rats were divided into two groups. The infrarenal abdominal aorta was infused with 15 U of elastase for 15 min (n = 12, 15-min infusion group) or 30 min (n = 12, 30-min infusion group). In each group, three rats were killed immediately and 1, 3, and 7 days after infusion, and then the aortas were excised for a histologic examination. Elastin fibers did not stain, even immediately after elastase infusion, in the 30-min infusion group. The degeneration of elastin fibers did not progress in the 15-min infusion group during the period of observation. Inflammatory cells infiltrated mainly to the adventitia near regions where the degeneration of elastin fibers spread totally through the aortic media. Elastin fibers degenerate immediately after elastase infusion and thus seem to degenerate not due to endogenous proteinases that are produced by the infiltrating cells, but due to the exogenously infused elastase itself. Inflammatory cell infiltration was thus found to be a result of the degeneration of elastin fibers in this model.

Animals↗

Biochemical changes in intervertebral disc degeneration.

The distribution of the principal matrix components, collagen, proteoglycans and water, across the diameter of human normal and degenerate intervertebral discs was compared. Little difference in collagen distribution was noted between normal and degenerate tissue but water and proteoglycan content decreased with degeneration, particularly in the centre of the disc. Proteoglycans of the nucleus pulposus and annulus fibrosus of normal and degenerate intervertebral discs were examined. In comparison with monomers of normal tissues, degenerate disc proteoglycans were of larger average hydrodynamic size and had a higher glucosamine to galactosamine ratio. Proteoglycans were digested with chondroitinase ABC and passed over an HS-Sepharose 2B affinity column. A greater proportion of the keratan sulphate-protein cores from degenerate disc were capable of interaction with the immobilized hyaluronate. Loss of aggregating ability was associated with diminution in size of the core. It is suggested that a large proportion of proteoglycans from normal disc have undergone a degree of degradation in the hyaluronate binding region and that proteoglycan synthesis in this tissue is slower than in degenerate tissue.

Adult↗

Regenerative effects of transplanting mesenchymal stem cells embedded in atelocollagen to the degenerated intervertebral disc.

Intervertebral disc (IVD) degeneration, a common cause of low back pain in humans, is a relentlessly progressive phenomenon with no currently available effective treatment. In an attempt to solve this dilemma, we transplanted autologous mesenchymal stem cells (MSCs) from bone marrow into a rabbit model of disc degeneration to determine if stem cells could repair degenerated IVDs. LacZ expressing MSCs were transplanted to rabbit L2-L3, L3-L4 and L4-L5 IVDs 2 weeks after induction of degeneration. Changes in disc height by plain radiograph, T2-weighted signal intensity in magnetic resonance imaging (MRI), histology, immunohistochemistry and matrix associated gene expressions were evaluated between normal controls (NC) without operations, sham operated with only disc degeneration being induced, and MSC-transplanted animals for a 24-week period. Results showed that after 24 weeks post-MSC transplantation, degenerated discs of MSC-transplanted group animals regained a disc height value of about 91%, MRI signal intensity of about 81%, compared to NC group discs. On the other hand, sham-operated group discs demonstrated the disc height value of about 67% and MRI signal intensity of about 60%. Macroscopic and histological evaluations confirmed relatively preserved nucleus with circular annulus structure in MSC-transplanted discs compared to indistinct structure seen in sham. Restoration of proteoglycan accumulation in MSC-transplanted discs was suggested from immunohistochemistry and gene expression analysis. These data indicate that transplantation of MSCs effectively led to regeneration of IVDs in a rabbit model of disc degeneration as suggested in our previous pilot study. MSCs may serve as a valuable resource in cell transplantation therapy for degenerative disc disease.

Aggrecans↗

Degeneration affects the fiber reorientation of human annulus fibrosus under tensile load.

The angled, lamellar structure of the annulus fibrosus is integral to its load-bearing function. Reorientation of this fiber structure with applied load may contribute to nonlinear mechanical behavior and to large increases in tensile modulus. Fiber reorientation has not yet been quantified for loaded non-degenerated and degenerated annulus fibrosus tissue. The objective of this study was to measure fiber reorientation and mechanical properties (toe- and linear-region modulus, transition strain, and Poisson's ratio) of loaded outer annulus fibrosus tissue using a new application of FFT image processing techniques. This method was validated for quantification of annulus fiber reorientation during loading in this study. We hypothesized that annulus fibrosus fibers would reorient under circumferential tensile load, and that fiber reorientation would be affine. Additionally, we hypothesized that degeneration would affect fiber reorientation, toe-region modulus and Poisson's ratio. Annulus fibrosus fibers were found to reorient toward the loading direction, and degeneration significantly decreased fiber reorientation (the fiber reorientation parameter, m(FFT)=-1.70 degrees /% strain for non-degenerated and -0.95 degrees /% strain for degenerated tissue). Toe-region modulus was significantly correlated with age (r=0.6). Paired t-tests showed no significant difference in the fiber reorientation parameter calculated experimentally with that calculated using an affine prediction. Thus, an affine prediction is a good approximation of fiber reorientation. The findings of this study add to the understanding of overall disc mechanical behavior and degeneration.

Anisotropy↗

Development of degenerate and specific PCR primers for the detection and isolation of known and putative chloroethene reductive dehalogenase genes.

Degenerate and specific PCR primers were designed for the detection of chloroethene reductive dehalogenases (CE-RDase), the key enzymes of chloroethene dehalorespiration, based on sequence information of three CE-RDases and three chlorophenol (CP) RDases. For the design of the degenerate primers, seven conserved amino-acid blocks identified with different bioinformatic tools were used. For one block degenerate, primers containing a 5'-consensus clamp region specific for CE-RDases and a 3'-end degenerate core region specific for RDases in general were designed using the Consensus-Degenerate Hybrid Oligonucleotide Primer (CDHOP) design method. Applying the degenerate primers to genomic DNA of Sulfurospirillum multivorans strain K, Dehalobacter restrictus strain PER-K23, and Desulfitobacterium sp. strain PCE1 led to the isolation of the known CE-RDase genes and three new genes encoding putative reductive dehalogenases that cluster with CE-RDases and not with CP-RDases. In addition, primers designed to be specific for the three known CE-RDase genes, namely pceA of S. multivorans, pceA of D. restrictus, and tceA of Dehalococcoides ethenogenes were successfully tested on genomic DNA of different chloroethene-dehalorespiring bacteria. Nested PCR using degenerate primers followed by a PCR with specific primers allowed a sensitive detection of only 10(2) copies per reaction.

Amino Acid Sequence↗

Adjacent segment degeneration and adjacent segment disease: the consequences of spinal fusion?

Given the number of spinal fusions performed annually, concerns have mounted over the potential for adjacent segment degeneration (radiographic changes of degeneration at levels adjacent to a spinal fusion) and adjacent segment disease (development of new symptoms correlating with adjacent segment degeneration). This article reviews documented evidence on adjacent segment degeneration and disease as it relates to cervical and lumbar arthrodesis. There appears to be an incidence of adjacent segment degeneration and disease after arthrodesis that may be related to natural degeneration or the adjacent fusion. It remains to be seen whether restoration of motion with disc arthroplasty will alter the rate of adjacent segment degeneration or disease.

Biomechanical Phenomena↗

Advances in gene therapy for intervertebral disc degeneration.

BACKGROUND CONTEXT: Gene therapy is a growing concept in many fields of medicine, and its potential applications are numerous. With a growing understanding of the molecular and cellular biology of intervertebral disc degeneration, alternatives to current treatment options are under investigation. Gene therapy offers an exciting new direction in the treatment of intervertebral disc degeneration, and potential targets of genetic alteration are being explored. PURPOSE: To describe and update the recent advances in research on gene therapy for the treatment of intervertebral disc degeneration. STUDY DESIGN/SETTING: Review of current research for the application of gene therapy as potential treatment for intervertebral disc degeneration. METHODS: Literature review. RESULTS: There is a growing body of research pertaining to the use of gene therapy as an adjunct or alternative to the current treatment options for intervertebral disc degeneration. In vitro studies have demonstrated that transfer of cDNA encoding growth factors to intervertebral disc cells can favorably modify their metabolic and biological functions. Additionally, initial in vivo studies have demonstrated successful transduction of growth factors to the intervertebral disc with confirmed upregulation of extracellular matrix synthesis. Investigators continue to explore the potential of gene therapy with several factors for the treatment of intervertebral disc degeneration. CONCLUSIONS: The potential of gene therapy to alter the course of intervertebral disc degeneration holds much clinical promise and continues to stimulate further investigations.

Animals↗

Intervertebral disc degeneration: the role of the mitochondrial pathway in annulus fibrosus cell apoptosis induced by overload.

Degeneration of the intervertebral disk (IVD) is a major pathological process implicated in low back pain and is a prerequisite to disk herniation. Although mechanical stress is an important modulator of the degeneration, the underlying molecular mechanism remains unclear. The association of human IVD degeneration, assessed by magnetic resonance imaging, with annulus fibrosus cell apoptosis and anti-cytochrome c staining revealed that the activation of the mitochondria-dependent apoptosome was a major event in the degeneration process. Mouse models of IVD degeneration were used to investigate the role of the mechanical stress in this process. The application of mechanical overload (1.3 MPa) for 24 hours induced annulus fibrosus cell apoptosis and led to severe degeneration of the mouse disks. Immunostaining revealed cytochrome c release but not Fas-L generation. The role of the caspase-9-dependent mitochondrial pathway in annulus fibrosus cell apoptosis induced by overload was investigated further with the use of cultured rabbit IVD cells in a stretch device. Mechanical overload (15% area change) induced apoptosis with increased caspase-9 activity and decreased mitochondrial membrane potential. Furthermore, Z-LEHD-FMK, a caspase-9 inhibitor, but not Z-IETD-FMK, a caspase-8 inhibitor, attenuated the overload-induced apoptosis. Our results from human samples, mouse models, and annulus fibrosus culture experiments demonstrate that the mechanical overload-induced IVD degeneration is mediated through the mitochondrial apoptotic pathway in IVD cells.

Animals↗

Histologic evidence of degeneration at the insertion of 3 rotator cuff tendons: a comparative study with human cadaveric shoulders.

We determined on histologic examination the degree of degeneration at the insertion of 3 rotator cuff tendons in 76 cadaveric shoulders, 17 of which had a partial tear of the supraspinatus. Fiber thinning, the presence of granulation tissue, and incomplete tearing of fibers, all evidence of degeneration, were quantified separately for each tendon. Among the shoulders that were intact on macroscopy, no significant difference in degeneration score could be found. In all 3 tendons degeneration was more prominent on the articular sides compared with the bursal sides (P < .0001). The degeneration score of partially torn supraspinatus was significantly higher than that of the intact tendons (P < .0001). The extent of granulation tissue, 1 criterion of degeneration, seemed to contribute mostly to this difference. Intrinsic degeneration occurred foremost in the articular side of the rotator cuff and might constitute the primary cause of rotator cuff tearing.

Adult↗

Apparent diffusion coefficients and T2 relaxation time measurements to evaluate disc degeneration. A quantitative MR study of young patients with previous vertebral fracture.

PURPOSE: To assess the suitability of apparent diffusion coefficient (ADC) measurements to evaluate degeneration processes of the vertebral disc and to compare the results with T2 relaxation time measurements in both degenerated and normal intervertebral discs. MATERIAL AND METHODS: Fourteen young patients (8.8-20.8 years old) who had had a vertebral compression fracture at least 1 year earlier, underwent MR studies with diffusion imaging in three orthogonal directions and T2 relaxation time measurements. ADC values and T2 relaxation times of both degenerated and normal intervertebral discs were compared to the values of 20 healthy young asymptomatic control subjects. RESULTS: In the degenerated discs of patients, the ADCx and ADCy values were decreased compared to earlier determined values of healthy controls. ADC values in the z-direction in degenerated discs did not differ significantly from the values of controls. T2 relaxation times were shorter in the degenerated discs of patients compared to the values of controls. The greatest changes in both these values were observed in degenerated discs followed by discs with normal signal intensity adjacent to primary trauma area and secondary trauma area. CONCLUSION: We suggest that decreased ADC values reflect the lost integrity of the intervertebral disc. ADC measurements at MR may prove sensitive depicting of early degenerative changes in vertebral discs.

Adolescent↗