Leukocytes, cell adhesion molecules and ischemic acute renal failure

Ischemic acute renal failure (ARF) is a common clinical syndrome, associated with high morbidity and mortality, for which there is no specific therapy. Polymorphonuclear neutrophils (PMN) recruited during reperfusion have been implicated as mediators of renal parenchymal injury in ischemic ARF. Leukocyte adhesion molecules appear to facilitate PMN recruitment in this setting. Complementary studies using monoclonal antibodies, antisense oligonucleotides and gene "knock-out" indicate that blockade of CD11/CD18 integrins and intercellular adhesion molecule-1 (ICAM-1) attenuates ARF in some experimental models of renal ischemia. These exciting observations may herald the development of novel anti-adhesion strategies for use in human disease.     

Serum levels of endothelial and neural cell adhesion molecules in prostate cancer

The purpose of this study was to describe the longitudinal development of running economy [defined as the oxygen uptake (VO2) at a submaximal running speed] in males and females from teenage to young adult age using data from the Amsterdam Growth and Health Study. Submaximal VO2 (in ml.kg-1.min-1) was measured in 84 males and 98 females while they ran on a treadmill at a constant speed of 8 km.h-1 for 6 min at three different treadmill slopes (0%, 2.5% and 5%). This test was carried out six times, on the same subjects at the ages of 13, 14, 15, 16, 21, and 27 years. The longitudinal development of running economy in males and females was analysed using a two-way analysis of variance for repeated measurements. At all three slopes, a significant decrease in VO2 with increasing age was found for both males and females, implying a significant increase in running economy for both sexes. Males showed significantly higher VO2 values than females at all ages measured and for all three slopes, suggesting that females have a significantly higher running economy than males. In order to make a better comparison of the VO2 of individuals of different sizes, allometric models were used; power function ratios were constructed in which body mass was expressed to an exponential power. Following this analysis the difference in submaximal VO2 and running economy between males and females appeared even larger.     

Expression of cell adhesion molecules and common acute lymphoblastic leukaemia antigen in hepatoblastoma

Hepatoblastoma is an embryonal tumour of the liver, which often contains tissue components with multidirectional differentiation. The occurrence of cell surface antigens in this tumour has not been studied systematically, and we therefore investigated 20 hepatoblastomas for the expression of common acute lymphoblastic leukaemia antigen (CALLA) and cell adhesion molecules (CAMs) in their different tissue components. Epithelial tumour cells of fetal differentiation contained E-cadherin. This protein did not occur in tumour areas with embryonal or mesenchymal differentiation. In contrast, immature embryonal and anaplastic cells expressed CALLA and the hyaluronate receptor (HCAM, CD44). Both fetal and embryonal areas stained irregularly positive for ICAM-1, which, in contrast, was not present on anaplastic cells. Immature fibrous tissue did not contain any of these molecules except for ICAM-1. However, some cells adjacent to, or enclosed in, osteoid were positive for HCAM and NCAM. Like small undifferentiated hepatoblastoma cells, primitive mesenchymal spindle-shaped cells also expressed CALLA, HCAM, and the polysialylated embryonic form of NCAM strongly. This last is not present on other epithelial or mesenchymal tumour cells. Hepatoblastoma cells of varying differentiation express distinct patterns of CAMs and CALLA. Our results give further insight into their histogenesis and cellular interactions and may explain their variable ability for invasive growth and formation of metastases.     

Changes in cervical fibronectin levels during pregnancy, labor and in the postpartum period

The authors analysed the fibronectin content of the uterine cervix during pregnancy and delivery. The aim of their investigation was furnishing data to the biochemical changes during cervical maturation. The ripening of the uterine cervix during pregnancy is a result of complicated interactions between different macromolecules, where the fibronectin plays a key role. The quantitative determination of the extracellular matrix fibronectin is impossible because its extraction from tissues recently is not solved. Taking this fact in consideration the authors choose a semiquantitative method, being reliable indicator of changes in fibronectin content of uterine cervix. They took small pieces of materials from portio vaginalis uteri of 139 women being in postmenopause and premenopause, in different stages of pregnancy and parturition concerning directly after delivery. The slides were incubated, with rabbit-anti-human fibronectin-FITC. The evaluation of fluorescence happened with an Axiophot (Zeiss) microscope. Authors stated that the fibronectin content in the cervical extracellular matrix and in the cellular membrane of fibroblasts increases during the 1st trimester pregnancy. This increase can be shown in the 3rd trimester as well and it drops significantly during delivery. They could not found any relationship between the leucocyte invasion observed during delivery and the changes of cervical fibronectin content. These observations call our attention to the importance of fibronectin in cervical ripening respectively dilatation and the need of further examinations.     

Multiple roles of neural cell adhesion molecule, neural cell adhesion molecule-polysialic acid, and L1 adhesion molecules during sensory innervation of the otic epithelium in vitro

To explore the role of cell adhesion molecules in the innervation of the inner ear, antibody perturbation was used on histotypic co-cultures of the ganglionic and epithelial anlagen derived from the otocyst. When unperturbed, these tissues survived and differentiated in this culture system with outgrowth of fasciculated neuronal fibers which expressed neural cell adhesion molecule and L1. The fibers exhibited target choice and penetration, then branching and spreading within the otic epithelium as individual axons. Treatment of the co-cultures, or of the ganglionic anlagen alone, with anti-neural cell adhesion molecule or anti-L1 Fab fragments produced a defasciculation of fibers but did not affect neurite outgrowth. In the co-cultures this defasciculation was accompanied by a small increase in the number of fibers found in inappropriate tissues. However, the antibodies did not prevent fiber entry to the otic epithelium. In contrast, removal of polysialic acid from neural cell adhesion molecule with endoneuraminadase-N, while producing a similar fiber defasciculation, also increased the incidence of fibers entering the epithelium. Nevertheless, once within the target tissue, the individual fibers responded to either Fab or to desialylation by spreading out more rapidly, branching, and growing farther into the epithelium. The findings suggest that fasciculation is not essential for specific sensory fibers to seek out and penetrate the appropriate target, although it may improve their tracking efficiency. Polysialic acid on neural cell adhesion molecule appears to limit initial penetration of the target epithelium. Polysialic acid as well as neural cell adhesion molecule and L1 function are involved in fiber-target interactions that influence the arborization of sensory axons within the otic epithelium.     

Changes in serum lipoprotein (a) levels related to hyperlipidemia during pregnancy--comparing normal pregnancy and toxemia of pregnancy

The aim of this 2-year study funded by the English National Board for Nursing, Midwifery and Health Visiting was to examine the relationship between teaching, support, supervision and role modelling in student clinical learning within the context of Project 2000 courses. The research, using a two-stage case-study design, employing predominantly qualitative methods of data collection, investigated the perceptions of students, tutors and practitioners of their experiences of these processes in the clinical setting. This paper focuses on the findings obtained from the practitioner data, high-lighting not only the significant role played by practitioners in facilitating student learning, but also the implications for practitioners in undertaking this role. The preparation required for supervising clinical learning, the integration of theory and practice and the organization of patient care have been identified as particularly important to the development of effective clinical learning environments.     

Changes in alpha (1-2)-fucosyltransferase activity in the murine endometrial epithelium during the estrous cycle, early pregnancy, and after ovariectomy and hormone replacement

The ratios of age-standardized mortality rates of Aboriginals to non-Aboriginals in Western Australia during the period 1983-1989 were 2.6 for males and 3.0 for females. Mortality rates experienced by Aboriginals were much higher in all age categories except 75+ years and for most major diseases except neoplasms. The peaks of all-cause age-specific mortality rate ratios (RR) for Aboriginal males and females were 10.2 (at 40-44 years) and 10.0 (at 35-39 years), respectively. These excess mortalities were mainly due to circulatory diseases, injury and poisoning, respiratory diseases and, in females, to digestive diseases and genitourinary diseases. The highest age-standardized, cause-specific RR for Aboriginal males were for mental disorders (10.3), injury and poisoning (8.9) and genitourinary diseases (8.6); for females the highest RR were for genitourinary diseases (16.9), endocrine, nutritional and metabolic (mainly diabetes mellitus) (12.3), and for infectious and parasitic diseases (7.5).     

Leishmania lipophosphoglycan reduces monocyte transendothelial migration: modulation of cell adhesion molecules, intercellular junctional proteins, and chemoattractants

We previously identified the structural requirement for the inhibitory activity of Leishmania lipophosphoglycan (LPG) to block endothelial adhesion to monocytes. Here we showed that LPG reduces transendothelial migration of monocytes. LPG pretreatment of endothelial cells (2 microM, 1 h) reduced monocyte migration across endothelial cells activated by bacterial endotoxin (LPS) or IL-1beta (60 and 46%, respectively). A fragment of LPG (i.e., repeating phosphodisaccharide (consisting of galactosyl-mannose)) and LPG coincubated with LPG-neutralizing mAb lacks inhibitory activity on monocyte migration. Pretreatment of monocytes with LPG (2 microM, 1 h) also did not affect monocyte migration through control or LPS-activated endothelial cells. FACS analysis reveals that LPG treatment blocked the LPS-mediated expression of E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 on endothelial cells and monocyte adhesion without altering the integrity of the endothelial monolayer. LPG (2 microM, 1 h) alone was capable of altering the expression and distribution of two junctional adhesion molecules, CD31 and vascular endothelium cadherin, as well as reversing the effects of LPS on these proteins. The induction of endothelial cells by LPS to transcribe and release monocyte chemoattractant protein-1 (MCP-1) was significantly reduced by LPG (40-65%). LPG treatment of nonactivated endothelial cells also suppressed by 55 to 75% the monocyte migration triggered by a MCP-1 chemoattractant gradient, and coincubation of LPG with neutralizing mAb abrogated the inhibitory activity. Together, these data point to a novel anti-inflammatory function of LPG in reducing monocyte migration across endothelial cells via a mechanism of inhibition of endothelial expression of cell adhesion molecules, modulation of intercellular junctional proteins, and synthesis of MCP-1.     

Neural cell adhesion molecules, learning, and memory in the domestic chick

The intermediate and medial hyperstriatum ventrale (IMHV) of the chick forebrain is a site of recognition memory for the learning process of imprinting. The results reported here demonstrate that neural cell adhesion molecules (NCAMs) play a time-dependent role in this recognition memory. Dark-reared chicks were trained, tested, and assigned a preference score as a measure of learning. Chicks with high preference scores were designated good learners and those with lower preference scores, poor learners. Controls were untrained. Tissue was removed, 9.5 hr or 24 hr after training, from the left and right IMHV, hyperstriatum accessorium, and posterior neostriatum. Three major NCAM isoforms (180, 140, and 120 kDa) were assayed. At 24 hr only, there was in left IMHV significantly more NCAM (for each isoform) in good learners than in the other 2 groups, and also a significant correlation between the amounts of NCAM and preference scores for all isoforms; the amount predicted by each regression line at preference score 50 (no learning) did not differ significantly from the mean value for untrained controls. There were no learning-related effects in either the hyperstriatum accessorium or the posterior neostriatum.     

Intracellular signaling is changed after clustering of the neural cell adhesion molecules axonin-1 and NgCAM during neurite fasciculation

Neural cell adhesion molecules of the immunoglobulin/fibronectin type III family on axons have been implicated in promotion of neurite outgrowth, fasciculation, and the mediation of specific cell adhesion. The present study demonstrates that two of these molecules on dorsal root ganglion neurons are associated with distinct protein kinases, axonin-1 with the src-related nonreceptor tyrosine kinase fyn and NgCAM with a casein kinase II-related activity and a serine/ threonine kinase related to S6 kinase. When neurites grew without contacts involving axonin-1 and NgCAM, strong fyn kinase activity was associated with axonin-1, whereas the NgCAM-associated kinase activities were low. Clustering of axonin-1 with NgCAM induced by the formation of cell-cell contacts correlated with a reduction of the axonin-1-associated fyn activity and an increased phosphorylation of NgCAM by the associated casein kinase II-related activity. Thus, axonin-1 and NgCAM trigger distinctive intracellular signals during in vitro differentiation depending on their state of association.     

Extensive platelet activation in preeclampsia compared with normal pregnancy: enhanced expression of cell adhesion molecules

OBJECTIVES: Platelets play an important role in the pathophysiologic mechanisms of preeclampsia. Our purpose was to investigate by means of flow cytometry to what extent platelets circulate in an activated state during normal pregnancy and whether this activation is more extensive in preeclampsia. STUDY DESIGN: Platelets in whole blood from 10 preeclamptic third-trimester pregnant women (highest diastolic blood pressure range 100 to 130 mm Hg, proteinuria range 0.59 to 11.5 gm/24 hr) and from 10 normotensive third-trimester pregnant controls were analyzed with the following activation markers: anti-P-selectin (alpha-granule secretion), anti-CD63 (lysosomal secretion), PAC-1 (monoclonal antibody against fibrinogen receptor conformation of the glycoprotein IIb/IIIa complex), anti-platelet endothelial cell adhesion molecule-1, and annexin-V (a placental protein that binds to negatively charged phospholipids, present on the outside of the platelet plasma membrane after activation). The differences in surface antigen exposure between the two groups were determined by double-label flow cytometry. Flow cytometric data were analyzed in two ways: first, the percentages of activated platelets above a certain threshold compared with a nonpregnant control sample were determined, indicative for activation of a subpopulation of cells, and, second, the mean fluorescence intensities were determined, indicative of the mean surface antigen expression of the total platelet population. RESULTS: Analysis of the percentage of activated platelets proved most informative. With this analysis an enhanced platelet activation status was present in 4 of 10 normotensive patients and a more extensive platelet activation status in all 10 preeclamptic patients, as indicated by P-selectin (p = 0.008) and CD63 (p = 0.03) expression. Increased platelet endothelial cell adhesion molecule-1 (p = 0.005) expression was also observed in preeclampsia. CONCLUSIONS: Flow cytometric analysis clearly indicated that platelets circulate in a more extensively activated state during preeclampsia than during normal pregnancy. The increased platelet endothelial cell adhesion molecule-1 expression in preeclamptic patients demonstrates that, besides alpha-granular and lysosomal release, other hitherto unknown mechanisms are involved. Platelet endothelial cell adhesion molecule-1 appears to be the best marker to distinguish preeclamptic patients from normotensive pregnant women. Only a subpopulation of the platelets appears to be activated.     

Expression of MHC and cell adhesion molecules in muscular sarcoidosis

Biopsied skeletal muscles from 5 patients with muscular sarcoidosis (nodular type; 1, and myopathic type; 4) were immunocytochemically examined. All biopsies presented granulomatous changes. Atrophic or regenerating muscle fibers adjacent to granuloma demonstrated compression or ischemic changes. In the center of the granuloma, CD68+ epitheloid cells and giant cells, and CD4+ T cells were localized. At the periphery of the granuloma, CD4+ T cells, CD8+ T cells, CD20+ B cells, and CD68+ macrophages were found. Expression of HLA-A,B,C was diffuse in the muscle fibers. Expression of HLA-DR and ICAM-1 was more prominent near the granuloma or perifascicular fibers, and that of LFA-3 was moderate in those lesions. VCAM-1 was expressed in endothelial cells and macrophages near the granuloma. Those findings indicate that interferon-gamma or TNF-alpha produced by infiltrating inflammatory cells may induce expression of these immunologic markers or adhesion molecules. Immunocytochemical differences between the nodular and myopathic forms of sarcoidosis are not evident, but either localization or abundance of granuloma in muscle bulks is relevant to weakness or atrophy of clinically affected muscle.     

Differential expression and distribution of focal adhesion and cell adhesion molecules in rat hepatocyte differentiation

The present study has examined the distribution of axons of differing sizes in the optic pathway of the ground squirrel. Axon diameters were measured from electron micrographs at various locations across sections of the optic nerve and tract, and total distributions and numbers were estimated. In both the nerve and tract, roughly 1.2 million optic axons were present. The population of optic axons had a unimodal size distribution, peaking at 0.9 microm in diameter and having an extended tail toward larger diameters. Local axon diameter distributions in the optic tract indicated distinct (though partially overlapping) axon diameter classes, including one of fine sizes peaking at 0.8-0.9 microm, a second of medium sizes peaking around 1.7-1.8 microm, and a third composed of the larger fibers with diameters up to 4.8 microm. The fine-caliber axons were found at all locations in the tract, and were the only axons present immediately adjacent to the pia, while the medium- and coarse-caliber axons were found at deeper locations. Curiously, the larger axons were found primarily in the medial parts of the tract, where axons from the dorsal retina normally course. A similarly restricted distribution of the larger axons was observed in the dorsotemporal parts of the optic nerve, suggesting that this difference in the tract may relate to an asymmetric distribution of ganglion cells on the retina giving rise to these axons. Measurements of axonal size taken within the optic fiber layer in dorsal and ventral parts of the retina confirmed this asymmetry, consistent with previous demonstrations of soma size differences in the dorsal versus ventral retina. The partial segregation of axons by size in the optic tract of the ground squirrel then reflects both the asymmetric distribution of retinal ganglion cell classes and the chronotopic reordering of optic axons that occurs within the chiasmatic region.     

Expression of T lymphocyte adhesion molecules: regulation during antigen-induced T cell activation and differentiation

The pattern of lymphocyte traffic and migration in vivo is a composite of constitutive recirculation and transient changes induced by interaction with antigen. Naive T lymphocytes in their basal, unstimulated state continuously recirculate throughout the entire host, poised to react to specific antigens that they are programmed to recognize. After interaction with antigen, T cell traffic changes, first with the trapping of reactive cells in antigen-containing lymphoid tissue. Subsequently, the effector cells responding to antigen, accompanied by nonspecific T cells and monocytes, traffic in large numbers to sites of antigen localization, resulting in the localized inflammatory response. Then, as the immune response wanes, memory T cells develop, many of which exhibit still different routes of recirculation. The traffic and tissue localization of leukocytes is regulated by a series of cell surface adhesion molecules that recognize specific ligands on endothelial cells and in the extracellular matrix. Modulation of the expression of these adhesion molecules results in the changes in T cell traffic that are characteristic of each stage of T cell differentiation. Thus, during T cell activation and differentiation, the down-regulation of adhesion receptors specific for lymphoid tissue endothelium and up-regulation of integrins facilitate the targeting of effector cells to sites of inflammation. Subsequent changes in adhesion receptors regulate the traffic of the antigen-specific memory cells. T cell adhesion molecule expression is therefore regulated as a function of the stage of activation and differentiation and, in addition, is influenced by cytokines and the local lymphoid microenvironment.     

Serum levels of soluble adhesion molecules in chronic renal failure and dialysis patients

The N-type voltage-operated calcium channel has been characterized over the years as a high-threshold channel, with variable inactivation kinetics, and a unique ability to bind with high affinity and specificity omega-conotoxin GVIA and related toxins. This channel is particularly expressed in some neurons and endocrine cells, where it participates in several calcium-dependent processes, including secretion. Omega-conotoxin GVIA was instrumental not only for the biophysical and pharmacological characterization of N-type channels but also for the development of in vitro assays for studying N-type VOCC subcellular localization, biosynthesis, turnover, as well as short-and long-term regulation of its expression. We here summarize our studies on N-type VOCC expression in neurosecretory cells, with a major emphasis on recent data demonstrating the presence of N-type channels in intracellular secretory organelles and their recruitment to the cell surface during regulated exocytosis.     

Plasma and intraerythrocyte amino-acid levels during and after haemodialysis in acute renal failure

A study is carried out on the variations of plasma and blood red cells free amino acid concentrations secondary to haemodialysis in patients suffering from acute renal failure. A reduction in plasma free amino acid pool has been observed in patients undergoing to many periodic haemosialysis, but no significant differences occur in plasma aminogram between before and after a single dyalitic procedure. Significant alterations were also observed in blood red cells aminogram, and this may be interpreted as reflex of intracellular omeostatic mechanisms for the maintenance of normal plasma free aminoacid pattern.     

Molecular biological approach to functions of telencephalin, a cell adhesion molecule and HNK-1 carbohydrate epitope, which is commonly expressed on cell adhesion molecules in the nervous system

Cell surface carbohydrates modulate a variety of cellular functions, including recognition and adhesion. The HNK-1 carbohydrate epitope, which is recognized by the monoclonal antibody HNK-1, is specifically expressed on a series of cell adhesion molecules and also on some glycolipids in the nervous system over a wide range of species from insect to mammal. The HNK-1 epitope is spatially and temporally regulated during the development of the nervous system and associated with neural crest cell migration, neuron to glial cell adhesion, outgrowth of astrocytic processes and migration of cell body, as well as the preferential outgrowth of neurites from motor neurons. These observations together with the unusual chemical nature of the HNK-1 epitope, namely a non-reducing terminal 3-sulfoglucuronic acid residue, prompted us to study the biosynthesis of the NHK-1 epitope, in which a unique glucuronyltransferase(s) plays a key role. We found that the respective glucuronyltransferases are involved in the biosynthesis of the HNK-1 epitope on glycoproteins (GlcAT-P) and on glycolipids (GlcAT-L). Then, we isolated a novel glucuronyltransferase (GlcAT-P) specific for glycoprotein substrates and its cDNA from the rat brain. The primary structure deduced from the cDNA sequence predicted a type II transmembrane protein with 347 amino acids. Transfection of the GlcAT-P cDNA into COS-1 cells induced not only expression of the HNK-1 epitope on the cell surface but also marked morphological changes of the cells, suggesting that the HNK-1 epitope was associated with the cell-substratum interaction. The GlcAT-P cDNA obtained in this study will be a useful molecular tool to open the way for further steps in the elucidation of the biological function of the HNK-1 carbohydrate epitope in the development of the nervous system.