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Biomedical subjects

N Shepard

Publications and source records attributed to N Shepard.

At least 19 recordsLinked to original sources

The nursing role in radiation oncology.

OBJECTIVES: To reflect on the historical evolution of radiation oncology nursing. DATA SOURCES: Published articles and texts, historical archives of professional organizations, and the authors' experience. CONCLUSIONS: The radiation oncology nursing role is multifaceted. Role components include patient assessment, patient education, support and counseling, physical care, continuity of care, research, and administrative and leadership activities. There is wide variation in staffing patterns and role implementation throughout the world, and in recent years there has been increasing interest in the development of advanced practice nursing roles in this specialized area of oncology. IMPLICATIONS FOR NURSING PRACTICE: Changes in the health care environment make it difficult to predict how the role of the nurse in radiation oncology will evolve in the future. Radiation oncology nurses will need to be proactive in developing the knowledge and skills needed to meet the challenges of the future.

Clinical Competence↗

Characterization of human type II procollagen and collagen-specific antibodies and their application to the study of human type II collagen processing and ultrastructure.

Type II collagen is the most abundant collagen in articular cartilage and, together with other tissue-specific collagens and proteoglycans, provides the tissue with its shock-absorbing properties and its resiliency to stress. Specific antibodies which recognize various collagen types have been very useful in the study of collagen biosynthesis, structure and metabolism in normal and pathological conditions. Antibodies which recognize epitopes of type II collagen have been described previously; however, many of these antibodies display cross-reactivity with other collagens or with type II collagen from other species, reflecting the high degree of homology of the helical domains of fibrillar collagens. In this study, we prepared antibodies to sequential determinants of human type II procollagen employing synthetic peptides with sequences deduced from the nucleotide sequence of the human alpha 1 (II) procollagen cDNA. The antibodies were highly specific for epitopes in either the C-terminal propeptide or the telopeptide of the human type II collagen and did not cross-react with other human interstitial collagens or with murine type II collagen. These antibodies were used in conjunction with biosynthetic labeling to study the secretion and processing of human type II procollagen and collagen in human chondrocytes in vitro. The results indicated that a lag period of about 90 min was required for the secretion of newly synthesized type II procollagen. Conversion of the secreted procollagen into fully processed alpha-chains and their deposition in the cell layer were first apparent 240 min following the initiation of biosynthetic labeling. The antibodies were also used to examine, by immunoelectron microscopy, the structure of the extracellular matrix produced by human chondrocytes maintained in long-term cultures under conditions which permit the preservation of the cartilage-specific phenotype. These highly specific antibodies provide valuable tools to study the metabolism and structure of human type II procollagen and collagen in normal and pathologic conditions.

Amino Acid Sequence↗

Postural control in young and elderly adults when stance is perturbed: kinematics.

Increased postural sway and falling are associated with aging and are likely related to problems with postural control in the elderly. We investigated the motions of individual body segments in 24 healthy young adults and 15 healthy elderly adults (mean ages 26 and 72) in response to four tasks: (a) standing with feet flat on an anteriorly accelerating platform (Flat Translation); standing on a narrow beam support that was (b) stationary (Beam Standing) and (c) accelerating anteriorly (Beam Translation); and (d) standing on a rotatable but otherwise stationary springboard (Springboard Standing). An optoelectronic camera system was used to measure rotations of body segments, particularly regarding their maximum excursions, time to first rotation response, direction of initial rotation, and time to first rotation reversal. In general, larger rotation excursions were noted in the elderly compared to the young group, particularly in the Beam Standing and Beam Translation tasks, but the magnitude of rotation difference was small. All rotation magnitudes were well within the available ranges of motion of the body joints. In both excursion magnitudes and directions of initial rotation, the elderly showed greater variability than the young. In the Beam Translation task, the elderly group, compared to the young, tended to rotate their upper body segments more than in the Flat Translation task. These data suggest that healthy elderly adults with no apparent musculoskeletal or neurological impairments have small but consistent differences in postural control kinematics, particularly when more challenging conditions are presented. Moreover, these data provide the basis for biomechanical analyses of joint torques and other dynamic requirements of these responses.

Adult↗

The clones of osteoarthritic cartilage.

We studied the early cartilage changes in osteoarthritis, examining the most normal appearing articular cartilage from the hips of 17 patients. Normal appearing cartilage from five patients treated for fractures was used as control material. Two different types of clone were found. The first had increased staining for proteoglycan and was thought to have been engaged in the synthesis of matrix. The other type was associated with a severe deficiency of proteoglycan, matrix streaks and evidence of degradation and phagocytosis of matrix components. Immunohistochemistry demonstrated large amounts of chondroitin 4 and 6 sulphate about the synthetic-type clones, and little or no reactivity about the degenerative clones which lay more superficially and were associated with matrix destruction. Clones appeared to be engaged in either matrix synthesis or its destruction. The disease process of osteoarthritis appeared to begin at the surface of the articular cartilage.

Aged↗

Altered coriolis stress susceptibility in essential hypertension.

Patients with hypertension frequently have vague complaints of dizziness and many other symptoms experienced by healthy individuals with motion sickness. We examined vestibular function in patients with essential hypertension, and we determined whether patients with essential hypertension are more prone to motion sickness using Coriolis stress testing. Vestibular function and Coriolis stress susceptibility were measured in 12 normotensive (NT) and seven asymptomatic patients with mild essential hypertension (HT). The Coriolis stress susceptibility index (CSSI) was calculated from the number of head movements in the four cardinal directions an individual could complete while being rotated in a computerized chair at increasing velocity before they developed motion sickness. The patients with hypertension had normal vestibular function and normal vestibuloocular responses as measured by standard techniques. Subjects with hypertension had significantly decreased Coriolis stress susceptibility scores compared to normotensive subjects (NT, 29.70 +/- 4.8; v HT, 5.48 +/- 2.0, P less than .001) and significantly decreased suppression of postrotatory nystagmus (NT, 44.5% +/- 3.8; v HT, 19.1% +/- 6.9, P less than .05). Medical treatment of hypertension did not result in an increased tolerance to provocative stimuli for motion sickness. It is suggested from our data that an increased susceptibility to motion sickness and abnormal vestibular responses to normal motion may account for many of the vague symptoms of "dizziness" reported by a large number of hypertensive patients.

Adult↗

Improved chondrocyte morphology and glycogen retention in the secondary center of ossification following osmium-potassium ferrocyanide fixation.

The use of osmium-potassium ferrocyanide as the secondary fixative greatly improved chondrocyte preservation and stabilized the cartilage matrix proteoglycan. The proteoglycan was similar in appearance to that seen following fixation in the presence of cationic dyes. Extensive glycogen preservation was noted in these cells, occupying the area prior to and during the formation of the secondary center of ossification. The volume and organization of the glycogen within the cell cytoplasm were greater than that following buffered osmium fixation, and the cellular vacuoles within were greatly reduced. The cells forming the secondary center prior to the onset of mineralization were of greatest interest, because other studies compared them with the primary growth plate and described them as showing signs of hypertrophy as early as 5 days postnatally, as is found in the primary growth plate. Our observations indicate that glycogen is present in these cells, and cellular enlargement was not present. The cells do not resemble the hypertrophic chondrocytes of the primary growth plate, as far as cytoplasmic content is concerned, and we suggest that they may contribute to the development of the secondary center in a different fashion.

Animals↗

The deleterious effects of drying on articular cartilage.

Knee joints of mature rabbits were exposed to room air for periods of time ranging from thirty minutes to one hour in an attempt to mimic the human situation in the operating room. The animals were killed after the joint had been closed and activity in a cage had been allowed for twenty-four hours. When the animal was killed, cartilage was removed and was incubated with radioactive proline for four hours before light microscopy autoradiographs were made. Other samples were prepared for study by electron microscopy. The results in the animals that were killed immediately after the cartilage was exposed to room air and in those that were killed twenty-four hours after closure of the joint were identical. Both the ultrastructural and the autoradiographic findings indicated that the entire thickness of the articular cartilage was necrotic after sixty minutes of drying. Forty-five minutes of drying produced complete necrosis of the cartilage in half of the animals. In the other half, some cells survived, although many areas of the cartilage had complete necrosis, top to bottom. Thirty minutes of drying produced patchy necrosis that extended only to the middle zone of the cartilage. In joints that were exposed to room air for one hour, necrosis of the chondrocytes was completely prevented by irrigating the joint every five minutes with Ringer lactate solution.

Air↗

Periodic acid Schiff--p phenylenediamine staining of glycogen in chondrocytes. A new combination which improves both cellular detail and glycogen identification.

This study reports a method whereby glycogen is identified in the chondrocytes of the secondary center of ossification prior to mineralization. The use of new fuchsin rather than basic fuchsin on one micron Spurr sections of femoral head cartilage fixed with potassium ferrocyanide-reduced osmium produced excellent identification of glycogen and when followed by p phenylenediamine, intensified cellular detail.

Animals↗

Mineralization in osteoblast cultures: a light and electron microscopic study.

Osteoblasts isolated mechanically from newborn mouse calvaria produced a calcified matrix when cultured in the presence of 10 mM beta-glycerophosphate or 3 mM inorganic phosphate. The uncalcified matrix revealed numerous matrix vesicles scattered among collagen fibrils. The calcified matrix showed mineralized collagen fibrils and calcified nodules whose underlying organic matrix was detected after decalcification. These structures resembled those described in fetal and woven bone. In partially decalcified areas, calcification was shown to spread out from these structures along collagen fibrils. Alkaline phosphatase activity was found associated with the plasma membrane and matrix vesicles. X-ray diffraction analysis demonstrated that the mineral phase deposited in culture was hydroxyapatite. These observations which demonstrate that the isolated cells elaborate in culture a mineralized matrix with chemical and ultrastructural properties of woven bone further support the osteoblastic nature of the cells.

Alkaline Phosphatase↗

Effect of patellar shaving in the rabbit.

Shaving of the articular cartilage on the undersurface of the patella was carried out in mature white New Zealand rabbits to determine whether the removed cartilage reformed or the shaving led to later evidence of degenerative arthritis. The animals were followed for as long as 15 months after surgery, at which time patellae were removed and examined by light and electron microscopy. The depth of shaving varied considerably; however, no evidence of repair and no evidence of degenerative arthritis was noted in either the superficial or deeply shaved patellae. It is concluded that such superficial cartilage defects neither repair or produce subsequent degenerative arthritis.

Animals↗

The antigen induced arthritis model: the relevance of the method of induction to its use as a model of human disease.

The much favoured ovalbumin antigen induced model of arthritis in rabbits is widely used in rheumatoid arthritis (RA) research. When examined histologically, it was found to have important deficiencies as parallels to the human disease. After sensitization to ovalbumin, 2 intraarticular challenge doses of a magnitude at each end of the spectrum used by investigators were used in 152 rabbits. The effects of the high and low dose challenges were examined histologically with particular attention to the articular cartilage. With high doses, the gross and histological changes in the knee joint were remarkably akin to acute cartilage necrosis rather than RA1. In the low dose, a milder smoldering arthritis was produced. These observations suggest that, depending on the challenge dose used, there is a tremendous variability in the kind of arthritis produced by the antigen induced arthritis model. Furthermore, it is suggested that previous conclusions about the pathophysiology and immunology of RA drawn from the models that produce a rapid and severe arthritis should be reexamined.

Animals↗

Ultrastructural modifications of proteoglycans coincident with mineralization in local regions of rat growth plate.

We carried out histochemical and electron-microscopic studies to examine the relationship between mineral deposition and changes in the organization of proteoglycans during mineralization in the cartilaginous growth plate of the rat. To preserve the distribution and organization of proteoglycans in the extracellular matrix and to stain proteoglycans, acridine orange was included in the solutions that were used for fixation and demineralization. In undecalcified sections, mineral crystals that initially appeared at the level of the penultimate hypertrophic chondrocyte grew into dense, spherical mineral clusters, roughly one micrometer in diameter, in the longitudinal septa of the lowermost hypertrophic zone. Following the removal of mineral with EDTA in the presence of acridine orange, proteoglycan structures with a characteristic rosette-like architecture were revealed within the matrix of the longitudinal septa of the lower hypertrophic zone. The location of these rosette-like proteoglycan structures was identical to that of the dense, spherical mineral clusters in the undemineralized controls, and they were remarkably similar in morphology and size. In the electron micrographs stained with uranyl acetate and lead citrate, the greatly increased density of the rosette-like structures and large diameter of the proteoglycan structures located in mineral clusters contrasted sharply with the slender strands of more faintly stained proteoglycan that was diffusely distributed throughout most of the extracellular matrix. X-ray microprobe analysis for sulphur confirmed the existence of proteoglycans in the rosette-like structures and demonstrated that the concentration of proteoglycans was selectively increased in these regions. The cores of the metaphyseal calcified cartilage also exhibited rosette-like proteoglycan structures, which before demineralization were totally obscured by the diffuse, dense homogeneous deposition of mineral. Without stabilization of the proteoglycans in the extracellular matrix with acridine orange, the rosette-like proteoglycan structures could not be demonstrated. These results clearly indicate that there is a selective increase in the concentration of proteoglycans in exactly the same regions where mineral clusters are formed in the hypertrophic zone of the cartilaginous growth plate.

Acridine Orange↗

The sulphur content of chondrocyte nuclei.

The sulphur content as a measure of glycosaminoglycan content of growth plate cartilage was determined by energy dispersive x-ray analysis on fresh freeze dried unstained, unfixed ultra thin sections of rat growth plate. In the resting and proliferative zones, quantities of sulphur were found in the nuclei equal to that of the matrix. Less sulphur was present in the cytoplasm. In areas of cell degeneration nuclear and cytoplasmic content of sulphur fell to levels a fraction of that seen in the matrix. It was presumed that most of the sulphur was in glycosaminoglycans. Although glycosaminoglycans have been reported in small amounts in the nuclei of cells, no study of the glycosaminoglycan content of chondrocyte nuclei has been reported. The use of freeze dried unstained, unfixed sections presumably prevented the migration of sulphur and glycosaminoglycans from compartment to compartment.

Animals↗

An intercellular network artefact in glycol methacrylate dehydrated neonatal cartilage.

A striking network thought to consist of glycosaminoglycan in neonatal cartilage has recently been reported following glycol methacrylate (GMA) dehydration and embedding. This study attempted to learn why the network was not seen in other preparations previously reported. Comparative light and transmission electron microscopy of the network structures was carried out to determine their biological significance. We have shown the networks to be reproducible artefact, attained only following GMA dehydration of glutaraldehyde fixed neonatal cartilage.

Acrylates↗

The measurement of proteoglycan in the mineralizing region of the rat growth plate.

UNLABELLED: Using a recently developed technique for measuring proteoglycan by x-ray microprobe analysis, we examined six-week-old rat growth plates. In the proliferating zone, the proteoglycan concentration was found to be relatively low in the transverse septa, but somewhat higher in the longitudinal septa between the proliferating chondrocytes. In the lower hypertropic zone, in the region of the degenerating hypertrophic chondrocytes, the immediate pericellular area had a very high concentration of proteoglycan, as was obtained. These results indicate that either proteoglycan concentrations fall to very low levels in fully mineralized cartilage, or that the mineralization of cartilage interferes in some way with the measurement of microprobe signals from proteoglycans. We speculate that high levels of proteoglycan are necessary for mineralization in the epiphysis of the rat. CLINICAL RELEVANCE: The spot-to-spot analysis of proteoglycan by microprobe demonstrates the usefulness of this tool in analyzing minute volumes of cartilage for proteoglycan. It further demonstrates that mineralization of growth-plate cartilage is associated with changing patterns of concentration of proteoglycan. These normal patterns may constitute a basis for comparison in similarly evaluating concentrations of proteoglycan in diseases of growth disturbance.

Animals↗