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Biomedical subjects

I M Shapiro

Publications and source records attributed to I M Shapiro.

At least 91 records · Page 5Linked to original sources
PubMed1988

Infection of a murine T-cell line with a retrovirus carrying c-myc decreases the interleukin-2 cell dependence by a nonautocrine mechanism.

A murine interleukin-2 (IL-2)-dependent T-cell line, CTLL-2, was infected with a retrovirus carrying the mouse c-myc cDNA and the Tn-5 neogene. Transduced cells were selected in the presence of Geneticin sulfate (G418); these cells were shown to express both the endogenous and the transduced c-myc genes. The IL-2 requirement of these cells was then found to be significantly reduced. The cells did not express IL-2 mRNA nor did they produce an activity mitogenic for CTLL-2 cells. This suggests that the reduction of IL-2 dependence observed following retroviral transduction of c-myc is caused by a nonautocrine mechanism.

Animals↗
PubMed1988

Linkage between energy status of perivascular cells and mineralization of the chick growth cartilage.

The objective of this investigation was to investigate the relationship between the energy status of epiphyseal chondrocytes of the chick growth cartilage and the development of mineralization. A microfluorimetric scanning technique was used to measure the reduced pyridine nucleotide content of transverse sections of freeze-trapped cartilage; these measurements were related to tissue structure by scanning electron microscopy. The results of this study show that the energy status of cells in the hypertrophic region of the growth cartilage is more complex than was previously believed. In hypertrophic cartilage, most chondrocytes are in a reduced state. However, in the early hypertrophic region, the vascular channels that penetrate the cartilage from the metaphysis exert a profound local effect on the energy metabolism of perivascular chondrocytes. Thus, around each of the channels, there exists a zone of chondrocytes about 40-60 micron wide which exhibits a low fluorescence yield. The fluorescence level suggests that these perivascular cells have a higher level of oxidative metabolism than hypertrophic chondrocytes that are a distance (greater than 150 micron) from the vascular channels. This finding, in conjunction with our earlier observation that mineralization is first seen in the perivascular region, leads us to the conclusion that mineralization is associated with cellular oxidative activity. We now reject the long-held concept that in cartilage the development of mineralization is entirely due to tissue hypoxia.

Animals↗
PubMed1987

Morphological observations concerning the pattern of mineralization of the normal and the rachitic chick growth cartilage.

The entire calcified layer of the chick growth cartilage is penetrated by canals that contains blood vessel complexes: some of these canals pass through all the layers of the cartilage from the resorptive zone at the metaphysis, through the mineralizing, hypertrophic, proliferative and resting regions. This study aimed to provide more details of the 3-D microanatomy of this region and to establish whether there are differences in the process and progress of mineralization compared with the established mammalian "model". Proximal tibial heads from 6 to 8 weeks old normal and vitamin D deficient chickens were rapidly frozen and prepared for scanning electron microscopy using freeze-fracture, freeze-drying, plasma ashing, and chemical deproteinization techniques. Cartilage samples were also embedded in PMMA and polished for BSE imaging. Other samples were prepared for light microscopy. Zones of (mineralized) cartilage several cells thick separate adjacent canals. At the mineralizing front, calcification of the matrix is most advanced close to the canals, but the matrix adjacent to the canal lumens does not calcify. Mineralisation of the cartilage matrix is incomplete and small fenestrae of unmineralized matrix connect chondrocyte lacunae. These discontinuities in matrix calcification could serve as a route for diffusion of nutrients, metabolites and dissolved gases. The calcified cartilage is more mineralized than the contiguous developing bone. Osteoblasts surrounded by bone were seen to occupy the lacunae of hypertrophic chondrocytes. We tentatively suggest that some osteoblasts represent a terminal stage in the differentiation of hypertrophic chondrocytes. The rachitic cartilage was disorganised. It was penetrated by irrugular vascular canals and exhibited a greatly expanded hypertrophic zone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Mineralization of normal and rachitic chick growth cartilage: vascular canals, cartilage calcification and osteogenesis.

This paper reviews recent work in the authors' laboratories that has led to new observations and thoughts concerning the mineralization of normal and rachitic chick growth cartilage. The proximal tibial growth cartilages of normal and rachitic chicks were rapidly frozen and prepared for SEM and biochemical studies. Using a scanning microfluorimetric technique we showed that at the mineralization front of normal and rachitic cartilage there is an abrupt change in chondrocyte metabolism. Thus cells in this region exhibited an increase in NADH and oxidative metabolism. In rickets, there was a decrease in the reduced pyridine nucleotide content of each of the zones. The reversal in chondrocyte metabolism was not due to low oxygen tension. SEM observations indicated that this region of cartilage was well supplied with vascular channels; moreover, mineral was first seen deposited in matrix in close proximity to the blood supply. Indeed these vascular channels appeared to be a basic architectural feature of normal cartilage, although disorganized in the rachitic state. The morphological studies also showed that gaps existed in the continuity of the mineral phase in normal cartilage. Although the rachitic cartilage does mineralize, discontinuities in the mineral distribution are much more severe, with the general failure of fusion of adjacent mineral clusters. These structures would serve as pathways for transport of nutritional factors and gases to chondrocytes that are distant from the vascular channels. Observation of hypertrophic cells reinforced the view that some osteoblasts represented a terminal stage in the maturation of chondrocytes.

Animals↗
PubMed1986

Redox studies of the epiphyseal growth cartilage: pyridine nucleotide metabolism and the development of mineralization.

The objective of this investigation was to examine the redox status of chondrocytes in normal and rachitic growth cartilages and to relate energy metabolism to cell maturation and the initiation of mineralization. The redox status was evaluated by chemical analysis and by microfluorimetric scanning of rapidly frozen, freeze-fractured tibial growth cartilages. In the normal epiphysis, the redox pattern of both avian and lagomorph cartilages were very similar. Thus, in the proliferative tissue zone the NAD/NADH ratio was high; in the hypertrophic zone, the cells appeared to be reduced. The sharp border between the two zones suggested that the redox shift may be associated with development of hypoxia. Induction of rickets resulted in a fivefold decrease in the total concentration of pyridine nucleotides in the proliferating and hypertrophic zones. Furthermore, the NAD/NADH ratio was profoundly disturbed. In the mineralizing zone, there was an accumulation of reduced pyridine nucleotide. Healing, initiated by administration of vitamin D to the rachitic birds, caused a rapid increase in NAD and NADH in all zones of the growth cartilage. It was concluded that vitamin D deficiency leads to changes in the energy metabolism of growth cartilage and that these changes were related to the defective mineralization of the rachitic tissue.

Animals↗
PubMed1986

Transfection of lymphoblastoid cells using DNA-loaded reconstituted Sendai virus envelopes: expression of transfected DNA and selection of transfected cells.

The American Burkitt's lymphoma cell line Loukes was cotransfected with cloned BamHI K fragment of EBV DNA and a vector pSV2neo. Reconstituted Sendai virus envelopes (RSVE) loaded with DNA were used for efficient gene transfer. Two cell lines have been obtained following culture in the presence of geneticin sulfate (G-418). Messenger RNA from both transfected DNAs was expressed during the whole period of observation, 42 days after transfection. This method provides a relatively simple and efficient means for selection of lymphoblastoid cells expressing a transfected gene.

Burkitt Lymphoma↗
PubMed1986

An x-ray fluorescence technique to measure in situ the heavy metal burdens of persons exposed to these elements in the workplace.

Assays of hair and body fluid concentrations may be valuable measurements of acute exposure to a heavy metal, but they do not provide insight into the total heavy metal intake when the intake is low and chronic. The use of an x-ray fluorescence technique (XRF) enables measurement of the long-term retention of various heavy metals in select tissues in vivo. XRF was used to measure the mercury content of head and bone tissue in 298 dentists with long-term exposure to mercury-containing amalgams. It was also used to evaluate the lead burden of persons suspected of having elevated lead exposure at the workplace, and to assay the lead levels in urban and rural children. These studies indicated that the x-ray fluorescence method of assaying heavy metals in vivo is noninvasive, safe, rapid, and sensitive to levels of many heavy metals that accumulate in human tissues.

Adult↗
PubMed1986

Induction of concanavalin A dose-dependent T-cell growth factor production by insertion of T-cell membrane components into the AKR thymic lymphoma BW 5147.

Sendai virus vesicles were used as vehicles for the insertion of various cell membranes into different cell lines. The transplantation efficiency was measured by using FITC-labelled concanavalin A (Con A) or monoclonal antibodies against the T-cell marker Lyt 2 and the major histocompatibility complex product H-2Db in a fluorescence-activated cell sorter. Results indicate that it is possible to transplant mitogen responsiveness to certain cell types. Con A stimulation of T-cell membrane transplanted BW 5147 showed that it is possible to induce a mitogen dose-dependent T-cell growth factor production. Consequently this method appears to be an attractive model for further study of the properties of the membrane structures involved in mitogen triggering of cells.

Animals↗
PubMed1985

Low levels of mercury inhibit the respiratory burst in human polymorphonuclear leukocytes.

The objective of this investigation was to examine the effects of low levels of Hg(II) on the respiratory burst of PMNs by monitoring O2 consumption, superoxide radical formation, and chemiluminescence. Hg(II) at concentrations of 10-100 ng/ml profoundly inhibited zymosan-stimulated human cells. This inhibition was immediate in onset and occurred with minimal loss of cell viability. Effects of Hg(II) on the PMN respiratory burst were compared with those of Sn, Pb, Se, Au, Ag and Cu. Only in the case of Ag and Cu did the inhibitory effects approach those of Hg. The results indicate that Hg(II) may serve as a specific inhibitor of components of the respiratory burst.

Adult↗
PubMed1985

Morphochemical analysis of phosphorus pools in calcifying cartilage.

The objective of this investigation was to measure phosphorus (P) levels in the epiphyseal growth cartilage and to relate pool sizes to chondrocyte maturation and tissue mineralization. To carry out these studies, we utilized a morphochemical technique that permitted measurements of insoluble mineral phosphate, soluble inorganic phosphate (Pi), low and high molecular weight phosphorylated macromolecules and lipid P in freeze-trapped histological sections. Analysis of the sections revealed that very low levels of P are present in pre-mineralized cartilage; at the mineralization front, a large increase in Pi is correlated with mineral formation. Moreover, with calcification of the cartilage, a decrease in the concentration of low molecular weight compounds was observed. It is suggested that these latter components may provide the initial source of Pi for the development of mineral. The results of the study support the view that metabolic regulation of P pool size may be a rate-limiting factor in the mineralization of cartilage.

Animals↗
PubMed1985

Isolation of matrix vesicles by isoelectric focusing in Pevikon-Sephadex.

We have investigated the use of an isoelectric focusing (IEF) technique for isolating and characterizing matrix vesicles. Focusing was performed on crude preparations of matrix vesicles isolated from collagenase digests of chick epiphyseal cartilage and purified by discontinuous sucrose gradient centrifugation. Crude and partially purified vesicle preparations were subjected to flat bed IEF in a slurry of Pevikon-Sephadex. Partially purified matrix vesicles focused as a narrow band (pI congruent to to 6.5). Alkaline phosphatase, solubilized from matrix vesicles, focused with a pl of 4.0-4.5. The IEF profile of matrix vesicles also differed from that of chondrocyte membranes. Thus, the membrane pls were congruent to to 5.4 and 6.6-7.8, respectively. The latter peak probably corresponded to the pl of the matrix vesicle preparation. This observation lends support to the view that vesicles originate from distinct regions of the chondrocyte membrane.

Alkaline Phosphatase↗
PubMed1985

Accuracy of the Welch Allyn AudioScope and traditional hearing screening for children with known hearing loss.

The Welch Allyn AudioScope is a new screening instrument that allows both audiometric testing at 500, 1000, 2000, and 4000 Hz at 25 dB hearing level and otoscopic examination. Sixty-eight patients between 3 and 9 years of age with normal hearing or a variety of hearing losses comprised the study population. Immediately after receiving an audiologic evaluation the subjects were screened with a traditional screening audiometer and two examinations with the AudioScope, performed in a clinic examining room by pediatricians and nurse practitioners unaware of the children's hearing status. The AudioScope was as accurate as traditional hearing screening in identifying children with normal hearing and those with hearing loss. Log-linear modeling demonstrated that the results of AudioScope testing were similar to those of traditional hearing screening. However, the false positive rate was higher at 500 Hz for the first AudioScope examination, suggesting the need for two AudioScope tests. The AudioScope was quick and easy to use, nonthreatening to the subjects, less expensive than a pure-tone screening audiometer, and does not require a sound-treated room.

Audiometry↗
PubMed1985

Morphological observations of mineralizing pericardium cardiac grafts.

Pericardial patch grafts were implanted in the hearts of young sheep for periods ranging from two to 120 days. Explants seven to 21 days old revealed the formation of a "pseudoneointima" (PNI) on the blood contacting surface of the pericardium. The PNI was more heavily mineralized than the pericardium. Mineralization was most intense on the blood contacting surface of the PNI and on the chamber surface of the pericardium. After three weeks of implantation, the PNI was much thinner and was organized into a thin fibrous capsule without any signs of mineralization. In the pericardium, mineral deposits were seen in fibroblasts. Moreover, cell-related mineralization was evident prior to calcification of the surrounding collagen matrix.

Animals↗
PubMed1984

Phosphate modulation of calcium transport by a calcium-phospholipid-phosphate complex of calcifying tissues.

The ionophoretic activities of a stable Ca-PS-Pi complex were examined in a three-compartment lipid-aqueous phase transport system. It was found that the complex transported Ca2+ at a slower rate than free PS. Analysis of the transport process, using an equilibrium partitioning model, revealed that Ca2+ binding to the complex was the rate-limiting step. This inhibition was overcome by the addition of exogeneous Pi. It was concluded that the complex acted as an ionophore and that the presence of coordinated Ca and P atoms facilitated Pi modulation of Ca2+ transport.

Animals↗
PubMed1984

Microdissection--elemental analysis of the mineralizing growth cartilage of the normal and rachitic chick.

The concentrations of elements in avian growth cartilage were studied by electron probe x-ray emission microanalysis (EDX). The cartilage was prepared for analysis by freezing, freeze-fracturing, freeze-drying, and carbon coating techniques. Cells and matrix fragments were removed from the tissue by microdissection with a tungsten needle in a scanning electron microscope (SEM) equipped with a real-time stereoscopic viewing facility. The samples were analyzed in the same SEM by EDX. Elemental analyses were performed on each fragment at a distance from the tissue sample, and hence background radiation due to the sample was eliminated. An important finding was that the intracellular potassium concentration of chondrocytes in calcified cartilage was similar to the levels in the premineralized zones. This observation supports the view that chondrocytes do not die in the process of, or as a consequence of, mineralization of the surrounding matrix. Calcium peaks were seen in the matrix at all levels and in chondrocytes immediately prior to mineralization. In contrast, phosphorus levels were always high in cells and low or absent from the premineralized matrix. At the mineralization front the appearance of a phosphorus peak in the matrix just preceded the deposition of mineral. We propose that the transfer of phosphorus from cell to matrix is a rate-limiting step in mineralization. Finally, when rachitic and normal cartilage were compared, little difference was seen in the profile of either intracellular or extracellular elements. However, in rickets the mineralized matrix remained soft in consistency. We suggest that this may reflect a phosphorus-related calcification defect that prevents growth and interlocking of the apatite crystallites.

Animals↗
PubMed1984

Mineralization of short term pericardial cardiac patch grafts.

Glutaraldehyde fixed patch grafts of bovine pericardium were implanted in myocardial windows in young (3-4 months old) sheep. The samples were retrieved after one to three weeks for study with scanning electron microscopy (SEM) and energy dispersive x-ray microanalysis (EDX). A layer of porous material (pseudoneointima, PNI), consisting mostly of a dense mesh of fibers interspersed with blood cells, was noted to form on the blood contacting surface of the graft. Four distinct sets of mineralization were noted in the retrieved grafts: (1) at the blood contacting surface of the PNI; (2) within the PNI at the junction between layers of PNI with differing densities; (3) near the junction of PNI and pericardium (but in the PNI); and (4) within the pericardium. In both the PNI and pericardium the mineral was shown by EDX analysis to contain both calcium and phosphorous indicating the mineral to be a calcium phosphate. Mineralization in the PNI differed from that in the pericardium; in the PNI it was deposited in discrete regions and apparently in association with thrombi while in the pericardium it was distributed diffusely within the collagen matrix, which may influence its formation.

Animals↗