Search

Chondroitin sulphate proteoglycans are synthesised by different tissues and cell types, and the chondroitin sulphate chains are variably sulphated. Three monoclonal antibodies 3B3, 7D4 and 6C3 that recognise different native chondroitin sulphate epitopes have been used to investigate changes in structure during embryonic tissue development in the chick and in the response of mature canine articular cartilage during experimental osteoarthritis. Strong focal expression of the epitopes was seen during development of chick bursa, which was different for the three epitopes and which changed during 5 days of development. In embryonic chick limb, although chondroitin sulphate is present throughout the cartilage, the 3B3 epitope, which is at the non-reducing terminus of chains, was only expressed on chondroitin sulphate within one region of the sub-articular cartilage. In mature canine articular cartilage the expression of this epitope on proteoglycans was very low, but when determined 3 or 6 months after induction of experimental osteoarthritis the level was greatly increased in all joints tested (23/23). The abundance of the other two native chondroitin sulphate epitopes was also increased in this experimental disease. The results show that expression of the chondroitin sulphate epitopes detected by the monoclonal antibodies changes during cellular differentiation and development and suggests that it is closely controlled by the cells synthesising chondroitin sulphate chains.

A 7-year-old boy developed rhabdomyolysis with a peak creatine phosphokinase level of 261,400 IU/L after his appendectomy. These abnormalities occurred following a 2-3-day illness consisting of upper respiratory tract symptoms, fever, and abdominal pain mimicking acute appendicitis. At the time of operation, a normal appendix was removed, and mesenteric lymphadenitis was noted. The myoglobinuria and elevation of creatine phosphokinase were transient, and the patient remained asymptomatic. We review various causes of right lower quadrant pain and rhabdomyolysis and address the roles of malignant hyperthermia and infectious agents. The possible cause of the phenomena observed in this patient is discussed.

Empirical studies on comprehension of information for informed consent show that research subjects often do not understand major portions of that to which they have consented. This raises the ethical question of whether or not the consent process has accomplished its purpose--the protection of the individual's right to self-determination. This article focuses on the following factors derived from the research literature that influence comprehension of information for informed consent: the nature of information, method of presentation of information, demographic factors, and personal factors. From these factors and methodological considerations, ethical implications for nursing research are addressed.

Resorption of uncalcified cartilage in the embryonic chick femur appears to be mediated by two types of mononuclear cells. One cell type lies flattened and adherent along the surface of the cartilage matrix into which it extends cellular processes. Cytological characteristics of a large, euchromatic nucleus containing a nucleolus, and cytoplasm containing moderate to extensive amounts of rough endoplasmic reticulum indicate that these are protein synthetic cells. Macrophages, characterized by a pleomorphic shape and cytoplasm containing numerous mitochondria and vesicles, comprise the second cell type. These may be seen lying in contact with cartilage matrix, but are more likely located in the nonhematopoietic marrow adjacent to resorbing cartilage, where they establish close cellular associations with protein synthetic cells. Alkaline and acid phosphatase histochemical studies differentiate these two cellular types. Marrow alkaline phosphatase activity is restricted to the cartilage-marrow interface from which it diffuses a short distance into cartilage matrix, but does not diffuse into nearby marrow. Intracellular alkaline phosphatase is present only in protein synthetic cells that line the surface of cartilage, and thus appears to be produced by these cells. Acid phosphatase positive macrophages are scattered throughout the marrow, but are found in greatest concentrations in the region of cartilage resorption. They are rarely in direct contact with cartilage, and there is no evidence that acid phosphatase is released from these cells. The relative localizations and the presence of cellular interactions of these two cell types suggests that protein synthetic cells may be of fibroblastic origin, and may play a primary role in cartilage degradation, while macrophages, in keeping with biochemical evidence, play an adjunct or possibly a regulative role.

Physicians in rural settings confront many of the same ethical dilemmas as their urban counterparts: confidentiality, quality-of-life decisions, resource allocation, and their moral responsibility for bettering the life of the community. However, the courses of action they choose as morally justifiable are influenced by distance from other professional facilities, the interrelationship of private and professional roles in a small community, and the non-specialized orientation of their practices.

Although lidocaine is commonly accepted as an anti-arrhythmic drug, it can also be used intravenously as an anesthetic agent. The author focuses on how lidocaine can be used as an integral part of the anesthetic itself for patients with cardiac arrhythmias, particularly premature ventricular contractions.

A panel of four separate monoclonal antibodies, all known to specifically recognize epitopes on keratan sulfate glycosaminoglycans, were employed in an immunocytochemical study of developing chick hind limbs. In addition, two monoclonal antibodies specific for epitopes on chondroitin/dermatan sulfate glycosaminoglycans were employed on equivalent sections to determine the degree of colocalization of keratan and chondroitin/dermatan sulfates. The spatial distributions of keratan sulfate and chondroitin/dermatan sulfate differed to some extent. In younger embryos, high extracellular concentrations of keratan sulfate occurred in joints and articular cartilages, with diminishing amounts being present in epiphyseal and diaphyseal regions. The high concentration of keratan sulfate in joints and articular cartilage corresponded to equally high concentration of chondroitin-6 sulfate. With advancing age, the above mentioned distribution was modified, most notably by increased amounts of keratan sulfate within diaphyseal regions. Finally, the use of four different anti-keratan sulfate monoclonal antibodies made it possible to compare keratan sulfate epitope expression. Differences in keratan sulfate epitopes were noted in some regions of bones, mostly in diaphyseal regions of younger bones and epiphyseal regions of older bones. This pattern of keratan sulfate expression suggests that different types of keratan sulfate may be present and their expression may be developmentally regulated.

During November and December 1981 more than 50 residents in a village in Derbyshire had an acute gastrointestinal illness. One month later a second outbreak occurred affecting another 22 people. Campylobacter jejuni was isolated from 12 patients; no other gastrointestinal pathogens were identified. A case-control study showed an association with the consumption of unpasteurized milk from one particular farm. No new cases were identified for 6 months following the application of a Pasteurization Order from 26 January to 23 February 1982.

9-[( 2-Hydroxy-1-(hydroxymethyl)ethoxy]-methyl)guanine (BW B759U) is a more potent inhibitor of human cytomegalovirus (HCMV) in vitro than is the related nucleoside analog acyclovir (ACV). BW B759U was selectively activated to the 5'-triphosphate (BW B759U-triphosphate) in cells infected with HCMV to levels at least 10-fold higher than those measured for ACV-triphosphate and up to as much as 100-fold higher than the levels found in uninfected cells. BW B759U-triphosphate accumulated in HCMV-infected cells with time; the rate of this increase was dependent upon the drug dose and virus multiplicity of infection. Enzyme activities that catalyzed the phosphorylation of thymidine and 2'-deoxycytidine increased 3- to 7-fold in extracts of cells early after HCMV infection but thereafter declined. No concomitant increase in the rate of BW B759U phosphorylation was detected under these assay conditions. Maximal rate of accumulation of both BW B759U-triphosphate and ACV-triphosphate after a short exposure to drug occurred in the late phase of the infective cycle, as the titer of extracellular virus reached a peak in untreated cultures, but after the decline of stimulated host deoxypyrimidine kinase activities. Once formed, the BW B759U-triphosphate pool decreased very slowly and thus it persisted for several days in both HCMV-infected and uninfected cells.

Light microscopic, scanning electron microscopic, and transmission electron microscopic studies of the early developmental stages of chick embryonic bone marrow disclose characteristic associations of the first hematopoietic cells with stromal cells. The first hematopoietic cells, large basophilic cells that we have termed presumptive stem cells, segregate into erythropoietic and granulopoietic regions. Intravascular erythropoietic cells associate with sinusoidal endothelial cells, while granulopoietic cells associate with extravascular reticular cells. Extensive, intimate contacts between erythroid and endothelial cells are maintained, in part, by marginal arrays of microtubules, which promote a flattening of the adherent erythroid cell surface. In addition, cell surface components of opposing cells, visualized by ruthenium red staining, appear to merge and possibly to interact. Granulopoietic cells establish intimate but less extensive associations with reticular cells through cell-surface interactions. Stationary granuloid cells appear to be held in place by small, thin processes emanating from the sheet-like reticular cells. Granuloid cells are capable of moving within the extravascular region, using reticular cell surfaces as a substrate. Intimate associations also occur among granulopoietic cells, the significance of which is unclear. Thus, sinusoidal endothelial cells and reticular cells comprise the critical non-hematopoietic or stromal elements of avian bone marrow, where they have a putative role in segregating presumptive stem cells into erythrocyteic and granulocytic compartments. They serve as an architectual, and possibly regulatory, framework on which hematopoiesis occurs.

Arterial development was studied in embryonic chick bone marrow starting on day 11 of incubation and continuing until day 18. Arteries first appear on day 11 as vessels with two components to their wall structure, namely a thin, complete endothelium and a nearly complete pericyte layer. With advancing age, three distinct layers--an endothelium, a complete muscularis, and adventitia--appear. Long, narrow endothelial cells form cellular junctions at their basal surfaces. Smooth muscle cells remain relatively undifferentiated and do not consist of more than three layers even in the largest arteries. The adventitia consists of a hypocellular zone containing fibroblasts and a collagenous extracellular matrix. Unmyelinated nerves are located at the peripheral margin of the adventitia. All of the larger arteries are surrounded by a sheath of heterophilic granulopoietic cells. These cells do not intrude into the adventitia, but basophilic or mast cells often do intrude. The stromal cells of the periarterial granulopoietic sheaths remain as either fibroblastic cells or as multilocular preadipocytes. They, unlike those stromal cells beyond the sheath, never become unilocular adipocytes. By day 18, granulopoiesis is restricted to the periarterial sheaths and to the endosteal regions of the marrow. These studies indicate that fibroblastic stromal cells and preadipocytes, but not fully developed adipocytes, support granulopoiesis in the chick marrow. The stromal cells in the periarterial sheaths either represent a subpopulation of cells that do not develop into adipocytes or represent cells whose development into adipocytes is locally inhibited.

Labelling by the galactose-specific lectin peanut agglutinin was studied in bone marrow of the embryonic chick at the electron-microscopic level by use of both a gold-conjugated lectin and an indirect, ferritin-conjugated, biotinylated lectin. Cell surface labelling is exclusively restricted to developing and mature heterophilic granulocytes, monocyte/macrophages, mast cells/basophils, all of which appear to develop and reside in the extravascular spaces of the bone marrow. Resident small lymphocytes, which comprise a minor portion of the cell population, are also labelled. Erythroid cells and thrombocytic cells, which develop inside venous sinusoidal vessels, display no labelling. The latter cells, like extravascular leukocytes, contain surface galactosyl residues located in subterminal positions on cell surfaces, since they are labelled by the galactose-specific Ricinus communis agglutinin-I. It is postulated that terminal galactosyl residues might be involved in interactions between the surfaces of extravascular leukocytes and extracellular matrix and/or stromal cell surfaces.

We investigated the utility of [3H]puromycin as an alternate and adjunct precursor to amino acids for measuring protein synthetic activity in rat liver slices. Slices were incubated in the presence of either [3H]puromycin or radiolabeled valine to compare the incorporation of these isotopic precursors into nascent hepatocellular proteins. Compared to liver slices from controls, comparable decreases in the incorporation of both [3H]puromycin and labeled valine were observed in experiments using slices from fasted rats and in slices preincubated with 25 mM ethanol. Radiolabeling of nascent polypeptides with either [3H]puromycin or labeled valine in liver slices from rats fed a liquid diet containing ethanol was also decreased compared to slices from pair-fed control and chow-fed animals. Our results demonstrated the validity of using [3H]puromycin to detect changes in protein synthetic activity under these conditions. The potential advantage of using [3H]puromycin for in vivo studies is discussed.

Sunday evening. You can't put off any longer the chore of correcting the stack of student papers. Picking up the first one, you scan it. It isn't good. The paper is full of sentence fragments and typographical errors. You decide that this is going to be a long evening. Scenes like this, hours spent correcting students' papers, only to discover that the students barely look at the comments, may lead faculty to avoid assigning writing. Fortunately, there are effective ways to respond to student writing without tying oneself down to hours of tedious correcting. These techniques can help nursing faculty enjoy reading and responding to students' papers, as well as help students to improve their writing skills.

The molecular weight of DNA from human peripheral lymphocytes has been measured on alkaline sucrose density gradients. The number average molecular weight (MN) of DNA is found to decrease as the age of the donor increases. This result is discussed with reference to lesions in DNA, both repairable and non-repairable, which may accumulate with age.

Lymphocytes from eleven patients with aplastic anaemia were cultured with various agents which interact with D.N.A., and the proliferative responses to phytohaemagglutinin were measured. Lymphocytes from seven of the eleven patients were unduly sensitive to bleomycin, an agent causing strand breaks in D.N.A. The findings suggest that D.N.A. in these seven patients was abnormal, possibly as a result of an abnormality of D.N.A. repair. It is suggested that in aplastic anaemia D.N.A. damage in stem cells may lead to a failure of proliferation.

Chrondroitin sulphate proteoglycans obtained from embryonic chick bone marrow and epiphyseal cartilage were compared using immunochemical and biochemical analyses. Proteoglycans from each tissue, separated on CsCl density gradients, under dissociative conditions, into high (1.6 g ml-1), medium (1.5 g ml-1) and low (1.4 g ml-1) buoyant density fractions, were immunochemically analysed, using a panel of monoclonal antibodies that specifically recognize chondroitin 4-/dermatan sulphates, chondroitin 6-sulphate, keratan sulphate, the hyaluronate binding region present on connective tissue proteoglycans, and link protein. The same antibodies were used in Western blot analyses to detect intact proteoglycan monomers and core proteins that had been fractionated by agarose-polyacrylamide and by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Specific differences between marrow and cartilage proteoglycans were detected. In CsCl gradients, marrow proteoglycans displayed a higher degree of heterogeneity in terms of buoyant densities and hexuronate distribution. Keratan sulphate chains were constituents of the majority of 'large' proteoglycans in the marrow; however, a portion of the large proteoglycans in marrow middle buoyant density fraction either lacked keratan sulphate chains or were substituted with a form different from that found on cartilage proteoglycans. Marrow lacked 'small' chondroitin/dermatan sulphate proteoglycans that were present in cartilage and contained a more heterogeneous population of proteoglycans, particularly in the lower buoyant density fractions. Both marrow and cartilage were similar in that they contained, as their major components, large, aggregating proteoglycans and link proteins that were immunochemically and biochemically identical. The significance of these differences between marrow and cartilage proteoglycans remains to be determined, but they may, in part, be responsible for imparting unique characteristics to the haematopoietic extracellular matrices.

Fibronectin was immunolocated in embryonic chick bone marrow by the use of both a direct peroxidase conjugated antiserum and an indirect Streptavidin bridge technique. Fibronectin is located in the extravascular granulopoietic compartment and, to a lesser extent, in the vascular, erythropoietic compartment. There is no evidence of fibronectin being associated with blood-stromal cell interactions involving either erythropoiesis or thrombopoiesis. However, mature thrombocytes display a substantial surface coat containing fibronectin. Much of the fibronectin appears to be situated on surfaces of those fibroblastic stromal cells which support granulopoiesis. Fibronectin containing extracellular material connects surfaces of developing granulocytes with surfaces of stromal cells. Fibronectin is a surface component of granulocytes as well as nearby stromal cells. However, there appear to be fewer ferritin particles per unit of surface on granulocytic cells. Many of the ferritin particles are not clearly associated with amorphous matrix material at cell surfaces. Immunocytochemical attempts to identify laminin were unsuccessful. These studies indicate that fibronectin is situated at sites where it could mediate adhesive interaction between granulopoietic cells and their stromal cells. Furthermore, cell surface-matrix interaction involving fibronectin could mediate migration of blood cells within the extravascular spaces.

It has long been known that uncalcified cartilage of embryonic chick long bones is removed to make way for invading marrow. However, no one has clearly established which cells are responsible for this erosion. Using the light and electron microscopes, we have studied the cartilage-marrow interface, which we presume to be the region of resorption. Here, we found two types of mononuclear cells in intimate contact with cartilage matrix. 1. The predominate cell type had a euchromatic nucleus with a nucleolus and a cytoplasm containing extensive profiles of rough endoplasmic reticulum; also, processes extended from these cells into the adjoining cartilage matrix. 2. Macrophages containing many lysosomal vesicles, which often became swollen, were found on or near the surface of cartilage. In addition, a few cells with an intermediate appearance were present. A decrease in the amount of sulfated material in a 25-30 micrometer zone of cartilage in advance of the interface and an alteration in the orientation, and in some cases the integrity, of collagenous fibers were associated with the presence of the above mentioned cells. These alterations in cartilage were not due to the synthesis of sulfated or of collagenous material. The above evidence, although not conclusive, suggests that these mononuclear cells are responsible for cartilage resorption. In this respect, the removal of avian uncalcified cartilage is similar to the resorption of uncalcified articular cartilage which occurs in rheumatoid arthritis.

Monoclonal antibodies directed against specific carbohydrate epitopes on chondroitin 4-/dermatan sulfate, chondroitin 6-sulfate, keratan sulfate, and a monoclonal antibody directed against the hyaluronate binding region were used to characterize proteoglycans extracted from embryonic chick bone marrow. About half of the proteoglycans separate into the high density fraction on a CsCl gradient. Glycosaminoglycan-specific antibodies recognize proteoglycans from all fractions; this includes an antibody directed against keratan sulfate. Some proteoglycans, principally in the high buoyant density fraction, contain sites recognized by the antibody specific for the hyaluronate binding region. Within limits of detection, all core proteins belong to the high-molecular-weight category, with weights in excess of 212 kD. Antibodies directed against chondroitin 4-/dermatan sulfate and against keratan sulfate primarily bind to extracellular matrix material located in the extracellular spaces and to matrix elements in the pericellular regions of fibroblastic stromal cells. The antibody that recognizes chondroitin 6-sulfate binds to sites on surfaces of fibroblastic stromal cells and also to extracellular matrix material. Little or no antibody binding is detected on surfaces of granulocytic cells. These studies indicate that chondroitin sulfate and keratan sulfate chains are both present in the proteoglycan extract.

Avian embryonic marrow is segregated into distinct erythropoietic and granulopoietic compartments. Within each compartment presumptive stem cells and immature blood cells establish intimate contact with their respective stromal cell. In this study we have examined one aspect of potential hematopoietic cell-stromal cell interaction by looking for the presence of intercellular junctions between these two elements. In previous studies, after aldehyde fixation, junctions were not observed, but after perfusion fixation with tannic acid-glutaraldehyde, pentilaminar junctions became evident. These junctions were most characteristically located in the intravascular erythropoietic compartment, but were also found in the extravascular granulopoietic compartment. Junctions frequently joined presumptive stem cells with sinusoidal endothelial cells as well as joining immature erythroblasts with sinusoidal endothelial cells; and less frequently, junctions connected adjacent erythroid cells. However, reticulocytes and erythrocytes were never seen to have formed junctions with any other type of cell. Similar junctions within the extravascular compartment connected contiguous reticular cells and also, on occasion, reticular cells with sinusoidal endothelial cells. Hematopoietic cell-reticular cell junctions were restricted to two classes of blood cells--extravascular presumptive stem cells and mast cells. There was no evidence of junctions connecting mature or maturing granuloid cells and any other cell type. The presence of intercellular junctions between immature blood cells and their respective stromal cells suggests that such interactions might play an important role in avian hematopoiesis. However, further work will be needed to determine if these junctions are merely adherence sites or whether they represent sites of intercellular communication. In either event, these junctions appear to reflect a mechanism whereby the marrow stroma could regulate erythroid maturation.

The sensitivity of human peripheral blood lymphocytes to bleomycin and mitomycin-C was assessed by measuring the inhibition of phytohemagglutinin stimulated proliferation. The sensitivity to bleomycin, and not to mitomycin-C, increased with the age.

We have isolated a human cytomegalovirus mutant that is resistant to the antiviral drug 9-([2-hydroxy-1-(hydroxymethyl)ethoxy]methyl)guanine (BW B759U), yet exhibits wild-type sensitivity to inhibitors of herpesvirus DNA polymerases such as phosphonoformic acid and aphidicolin. Cells infected with the mutant accumulate approximately equal to 1/10th the amount of drug triphosphate as do those infected with the wild-type parent. This reduction in drug triphosphate could not be attributed to altered drug uptake or to reduced stability of the triphosphate, once formed. The induction of normal nucleoside and deoxynucleoside triphosphates and certain cellular nucleoside kinases was comparable in mutant and wild-type virus infections. These results provide strong evidence for the importance of phosphorylation in the selectivity of this antiviral compound and raise the possibility that human cytomegalovirus encodes a nucleoside kinase. The mutant may identify the existence of a cytomegalovirus function whose properties could facilitate genetic analysis of this important pathogen.