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PubMed1975

Significance of elevated liver alkaline phosphatase in serum.

The serum alkaline phosphatase was fractionated by polyacrylamide gel electrophoresis in 317 patients with elevated serum alkaline phosphatase activity. In 253 patients the source of the elevation was the isoenzyme of presumed liver origin, band L. In 87 of these patients, there was either no obvious liver disease or the alkaline phosphatase elevation was inappropriately high. In 19 of the 87, liver disease was further excluded by liver biopsy or by laparotomy. Because of this, biochemical studies were done to verify the hepatic origin of band L. Band L and alkaline phosphatase extracted from human liver migrated together on polyacrylamide gel electrophoresis before and after digestion with Vibrio cholerae neuraminidase. They had identical pH optima, sedimentation coefficients, Michaelis constants, and rates of inactivation at 55.5 degrees C. They had different rates of inactivation in 3 M urea. Over-all, the data indicate that band L is of liver origin, and that elevation of the hepatic alkaline phosphatase isoenzyme may be a nonspecific finding in certain patients.

Alkaline Phosphatase↗
PubMedDate not supplied

Migration inhibitory factor-related protein (MRP)8 and MRP14 are differentially expressed in free-electron laser and scalpel incisions.

Incisions made in mouse skin by scalpel or the free-electron laser heal at different rates. To identify genes that are differentially expressed in free-electron laser or scalpel wounds, we isolated total RNA from free-electron laser- or scalpel-produced incisions and normal skin at day 7 postwounding. cDNA microarray analysis identified 89 of 15,000 genes in a mouse microarray as having significantly different expression levels. Migration inhibitory factor-related protein (MRP) 14 was almost 30 times more highly expressed in scalpel wounds than in free-electron laser wounds. This result was confirmed by Northern blot analysis, which also showed that scalpel wounds expressed higher levels of MRP8, a related S100 protein that can heterodimerize with MRP14, at days 2, 7, and 14 postwounding. Free-electron laser wounds also showed elevated expression of MRP8 and MRP14 relative to normal skin. In situ hybridization showed that the patterns of MRP14 and MRP8 expression in free-electron laser and scalpel wound tissues were similar. MRP14 and MRP8 were expressed in the dermal wound margin, while a very low level of MRP14 and MRP8 expression was seen in the migrating epidermis. Dual immunofluorescence staining for MRP14 or MRP8 and macrophage (F4/80) showed that most of the wound macrophages simultaneously expressed MRP14 and MRP8. Some expression was also found in neutrophils, while neither antigen accumulated to a significant degree in the epidermis. Relatively lower MRP8 and 14 expression in free-electron laser wounds was correlated with a higher level of matrix metalloproteinase-13 expression and a reduced rate of wound healing. While the regulation of MRP8 expression in mouse may be different from human skin, we suggest that elevated expression of MRP8 and MRP14 may have a relevant therapeutic effect against inflammation in wound healing.

Animals↗
PubMed2003

Interleukin-8 serum levels in patients with hepatocellular carcinoma: correlations with clinicopathological features and prognosis.

PURPOSE: In this study, we measured the serum interleukin-8 (IL-8) levels in patients with hepatocellular carcinoma (HCC) to evaluate its correlation with clinicopathological features and prognosis. EXPERIMENTAL DESIGN: ELISA was used to detect the concentrations of IL-8, vascular endothelial growth factor, and macrophage migration inhibitory factor in preoperative sera of 59 patients with resection of HCC and 15 healthy subjects. RESULTS: Preoperative serum IL-8 was found to be significantly elevated in patients with HCC compared with healthy subjects (median, 17.6 versus 1.0 pg/ml, P=0.046). The levels of IL-8 correlated significantly with a large tumor size (>5 cm), absence of tumor capsule, presence of venous invasion, and advanced pathological tumor-node-metastasis stage. Serum IL-8 level was a significant prognostic factor in terms of disease-free and overall survival. Patients with a serum IL-8 level of >17.6 pg/ml had a poorer disease-free survival than those with a level of <17.6 pg/ml (median disease-free survival 4.7 versus 19.2 months). Multivariate analyses showed that serum IL-8 level was a significant and independent prognostic factor of survival. CONCLUSIONS: Significant correlations of serum IL-8 levels with tumor size and tumor stage suggest that IL-8 may be directly or indirectly involved in the progression of HCC. These findings indicate that serum IL-8 may be a useful biological marker of tumor invasiveness and an independent prognostic factor for patients with HCC.

Adolescent↗
PubMed2006

Synthetic D-amino acid peptide inhibits tumor cell motility on laminin-5.

Cell motility is partially dependent on interactions between the integrins and the extracellular matrix. Our previous studies have identified synthetic D-amino acid cell adhesion peptides using a combinatorial screening approach. In this study, we demonstrate that HYD1 (kikmviswkg) completely blocks random haptotactic migration and inhibits invasion of prostate carcinoma cells on laminin-5. This effect is adhesion independent and reversible. The inhibition of migration by HYD1 involves a dramatic remodeling of the actin cytoskeleton resulting in increased stress fiber formation and actin colocalization with cortactin at the cell membrane. HYD1 interacts with alpha6beta1 (not alpha6beta4) and alpha3beta1 integrins and surprisingly elevates laminin-5-dependent intracellular signals including focal adhesion kinase, mitogen-activated protein kinase kinase and extracellular signal-regulated kinase. HYD1 does not contain a previously characterized binding sequence for integrins. A scrambled derivative of HYD1, called HYDS (wiksmkivkg), does not interact with the alpha6 or alpha3 integrin subunits and is not biologically active. Taken together, these results indicate that HYD1 is a biologically active integrin-targeting peptide that reversibly inhibits tumor cell migration on laminin-5 and uncouples phosphotyrosine signaling from cytoskeletal-dependent migration.

Actins↗
PubMed2005

[Reliability of porous coating metal-backed cups: advantages and adverse effect of a posterior elevated rim polyethylene liner].

PURPOSE OF THE STUDY: We analyzed primary implantation of the Duraloc cup associated with a self-locking Muller stem at 9.6 +/- 1.6 years (8.6-11.4). Because of its particular 3/5 semi-spherical shape, the Duraloc cup has been associated with a high rate of dislocation. The objective of this study was to evaluate the efficacy of an anti-dislocation posterior elevated rim polyethylene liner on long-term dislocation rate and wear. MATERIAL AND METHODS: The series included 89 arthroplasties implanted between 1991 and 1993 in 82 patients, mean age 56.8 +/- 12.6 years (17.2-87). The Postel-Merle-d'Aubligné (PMA) score and subjective evaluation with a visual analog scale were used for the clinical assessment. Changes in the bone-cup interface, cup migration, and polyethylene wear were assessed radiographically. RESULTS: At last follow-up, the PMA function score was 16.2 +/- 1.9 points (10-18) (81% good, very good and excellent results). The postoperative x-rays showed a low incidence of lucent lines and osteolysis, respectively 8% and 4%. Mean annual linear polyethylene wear was 0.11 +/- 0.066 mm (0.03-0.57) and only 4% of the cups showed wear greater than 0.2 mm/year. Wear was correlated with the presence of acetabular osteolysis and high activity level. Two cups migrated (3-4 cm medialization). Three cups were revised, one for deep infection, one for recurrent dislocation, and one for dislocation associated with major wear. The rate of dislocation was 2.2% (two cases). Overall survival was 97.3% at 9.6 years (95%CI 0.93-1.0). DISCUSSION: Compared with other series in the literature using this implant, the presence of a posterior elevated rim polyethylene liner reduced the rate of instability and did not increase wear. The Duraloc cup is recognized as a reliable implant exhibiting excellent osteointegration and a low rate of migration. The presence of a posterior rim is associated with a lower rate of dislocation and does not increase wear. It can thus be proposed for primary implantation. The posterosuperior position for the liner is recommended.

Adolescent↗
PubMed2006

The alpha1 subunit of GABAA receptor is repressed by c-myc and is pro-apoptotic.

The c-myc oncoprotein plays a critical role in the regulation of cellular proliferation and apoptosis. To mediate these biological functions, a variety of target genes are activated or repressed by c-myc, but few genes have yet been identified that directly mediate c-myc's role in proliferation or apoptosis. During a screen for genes that are repressed by c-myc, we identified the alpha1 subunit of gamma aminobutyric acid receptor (GABAAR-alpha1) as a novel target of c-myc. GABAAR is the major inhibitory neurotransmitter receptor in the mammalian central nervous system and is involved in developmental events in the brain, such as neurite outgrowth, neuronal survival, neuronal migration, and proliferation. We show here that GABAAR-alpha1 expression is rapidly and directly repressed by c-myc. GABAAR-alpha1 expression is elevated in c-myc null cells and upregulation of GABAAR-alpha1 correlates with downregulation of c-myc protein expression during neuronal cell differentiation. We also show that overexpression of GABAAR-alpha1 causes apoptosis, which is blocked by the coexpression of Bcl-2 or Bcl-XL. Induction of apoptosis is specific for the alpha1 subunit, since neither the beta1 or beta2 subunits of GABAAR induced apoptosis. Derepression of GABAAR-alpha1 expression upon downregulation of c-myc represents a unique apoptotic mechanism and a distinct function for the alpha1 subunit, independent of its role as a component of the GABAAR in the plasma membrane. In addition, the regulation of GABAAR-alpha1 expression by c-myc provides a potential direct role for the Myc proteins in neurological processes and neurodegenerative disorders.

Animals↗
PubMed1981

Creatine kinase BB and other markers of prostatic carcinoma.

Creatine kinase BB has been described to be elevated in the serum of patients with prostate cancer. The incidence of abnormal serum levels correlates with the degree of differentiation of the tumor; that is, the more poorly differentiated tumors are associated most frequently with elevated CK-BB levels. We have recently described two additional findings which involve CK-BB in prostate cancer: 1) the finding of abnormally migrating CK-BB bands reported to be IgG-CK-BB complexes in several Stage D patients and in 2/3 Stage B patients following 125I implants, and 2) significant differences between CK-BB purified from prostatic fluid from CK-BB of brain origin. In particular, we find two bands of prostatic CK-BB following purification as compared to a single band from brain, when the same purification protocol is used. While further characterization of these differences is proceeding, the two bands of CK-BB are being used to develop a prostate-specific antiserum for use in immunoassays to detect prostate cancer in conjunction with other traditional markers such as prostatic acid phosphatase. If serum detection of tumor markers is to be efficacious, tumor marker panels such as CK-BB and prostatic acid phosphatase may provide a significant advantage over individual markers.

Brain↗
PubMed1993

Properties of horizontal cells transiently appearing in the rat dentate gyrus during ontogenesis.

In an ontogenetic study, combining morphological analysis and patch clamp recordings, a transiently appearing horizontal cell type was identified in the dentate gyrus. The cells were exclusively located in the outer third of the stratum moleculare. They were present at postnatal day 2 (P2) and could be identified with fluorescent dyes until around P14. The morphology was bipolar, with a putative axonal and a dendritic process stretching out parallel to the pial surface without any preferential direction. Patch clamp studies in the current and voltage clamp mode were performed in hippocampal slices on visually identified horizontal cells, between P4 and P7, which were subsequently stained with lucifer yellow. The cells had a low resting membrane potential, around -55 mV. They were excitable, displaying broad action potentials (duration 3-20 ms) and, unlike mature dentate granule cells, they also expressed a strong delayed inward rectifier with properties reminiscent of the IQ current. Unlike granule cells, no postsynaptic signals could be observed during elevation of [K+]o or electrical stimulation, suggesting that the horizontal cells did not participate in functional hippocampal circuitry. We suggest that these cells represent migrating cells with subsequent differentiation to granule cells or inhibitory interneurons. Alternatively they may be part of the early radial glia or serve as transient target cells for afferent fibres between the entorhinal cortex and the dentate gyrus.

Animals↗
PubMed1988

In vitro evaluation of caval filters.

This experiment demonstrated the clottrapping ability of two commercially available filters, the Mobin-Uddin and Greenfield, and three experimental filters developed by Amplatz, Günther, and Gianturco. Each filter was tested in a polyethylene tube simulating the inferior vena cava. Separate series of 10 clots, each 3 cm long and 6 mm or 9.2 mm in diameter, were exposed to the test filter. The Mobin-Uddin and Amplatz filters failed by overload: acutely elevated pressures forced clots outside the skirt of the former, and between the limbs of the latter. The Günther filter trapped all incident clots, but migrated downstream when occluded by clot. The Greenfield filter passed clots at normal pressures between its widely spaced legs. The Gianturco (bird's nest) passed clots at normal pressures as well. While filters performed suboptimally, strengthening the anchoring struts of the Günther filter would result in a secure, effective filter.

Animals↗
PubMed2006

YKL-40, a new inflammatory marker with relation to insulin resistance and with a role in endothelial dysfunction and atherosclerosis.

Substantial evidence supports a role of chronic subclinical inflammation and activation of the innate immune system in the pathogenesis of insulin resistance and endothelial dysfunction and the development of type 2 diabetes (T2D) and atherosclerosis. Several proinflammatory cytokines, acute phase-reactants and cell adhesion molecules play a pivotal role in this chronic subclinical inflammation but a comprehensive understanding of the interrelations of these molecules is still needed. YKL-40 is a new inflammatory marker with relation to acute and chronic inflammation as well as cancer. It is secreted in vitro from a variety of human cells, including vascular smooth muscle cells (VSMCs), activated macrophages and macrophages during late stages of differentiation and is found in vivo in subpopulations of macrophages in tissues with inflammation and extracellular tissue remodelling, such as macrophages in atherosclerotic plaques. YKL-40 promotes chemotaxis, cell attachment and migration of VSMCs and the formation of branching tubules suggesting that YKL-40 plays a role in angiogenesis. Latest studies reveal that YKL-40 is elevated in patients with T2D and is related to insulin resistance. This article reviews the studies of YKL-40 with focus on a possible role of YKL-40 in insulin resistance, endothelial dysfunction and atherosclerosis.

Adipokines↗
PubMed2001

Expression of a novel member of the ATP1G1/PLM/MAT8 family, phospholemman-like protein (PLP) gene, in the developmental process of mouse cerebellum.

We have identified a new member of the ATP1G1/PLM/MAT8 family, named phospholemman-like protein (PLP), from a mouse cerebellum cDNA library. The family consists of small transmembrane proteins that modulate the activities of some ion channels. The deduced amino acid sequence of PLP consists of 93 residues that contain the ATP1G1/PLM/MAT8 motif and a single transmembrane domain, and is most similar to the sequence of mouse phospholemman. In situ hybridization analysis showed that the PLP gene is highly expressed in cerebellar granule cells. PLP expression is elevated in the postnatal developing cerebellum. Thus, it may be implicated in the proliferation, differentiation, and axon elongation of granule cells as they mature and migrate to the internal granule layer.

Amino Acid Sequence↗
PubMed2005

Neutrophil chemotaxis in a patient with neonatal-onset multisystem inflammatory disease and Muckle-Wells syndrome.

BACKGROUND: Neonatal-onset multisystem inflammatory disease (NOMID)/chronic infantile neurologic, cutaneous, and articular syndrome is an autoinflammatory disease characterized by urticarial rash, arthropathy, and central nervous system inflammation. OBJECTIVE: To describe a 13-year-old girl with overlapping symptoms of NOMID and Muckle-Wells syndrome who has a mutation in cryopyrin (NALP3). METHODS: We examined neutrophil migration using transwell assay and time-lapse videomicroscopy. We also examined p38 mitogen-activated protein kinase (MAPK) activation in patient and control neutrophils using Western blot analysis. RESULTS: Neutrophil defects in chemotactic migration were found to a variety of chemoattractants, including interleukin 8, N-formyl-methionyl-leucyl-phenylalanine, complement C5a, and leukotriene B4. Her neutrophils exhibited elevated basal and stimulated p38 MAPK activation in response to interleukin 8, N-formyl-methionyl-leucyl-phenylalanine, complement C5a, and leukotriene B4. CONCLUSIONS: This study is the first, to our knowledge, to demonstrate defects in neutrophil chemotaxis and p38 MAPK signaling in a patient with NOMID and Muckle-Wells syndrome and a cryopyrin mutation.

Adolescent↗
PubMed2004

Phenotypic conversion of human mammary carcinoma cells by autocrine human growth hormone.

We report here that autocrine production of human growth hormone (hGH) results in a phenotypic conversion of mammary carcinoma cells such that they exhibit the morphological and molecular characteristics of a mesenchymal cell, including expression of fibronectin and vimentin. Autocrine production of hGH resulted in reduced plakoglobin expression and relocalization of E-cadherin to the cytoplasm, leading to dissolution of cell-cell contacts and decreased cell height. These phenotypic changes were accompanied by an increase in cell motility, elevated activity of specific matrix metalloproteinases, and an acquired ability to invade a reconstituted basement membrane. Forced expression of plakoglobin significantly decreased mammary carcinoma cell migration and invasion stimulated by autocrine hGH. In vivo, autocrine hGH stimulated local invasion of mammary carcinoma cells concomitant with a prominent stromal reaction in comparison with well delineated and capsulated growth of mammary carcinoma cells lacking autocrine production of hGH. Thus, autocrine production of hGH by mammary carcinoma cells is sufficient for generation of an invasive phenotype. Therapeutic targeting of autocrine hGH may provide a mechanistic approach to prevent metastatic extension of human mammary carcinoma.

Autocrine Communication↗
PubMed2003

Insect resistance to Bacillus thuringiensis: alterations in the indianmeal moth larval gut proteome.

Insect resistance to the Cry toxins of Bacillus thuringiensis (Bt) has been examined previously using a number of traditional biochemical and molecular techniques. In this study, we utilized a proteomic approach involving two-dimensional differential gel electrophoresis, mass spectrometry, and function-based activity profiling to examine changes in the gut proteins from the larvae of an Indianmeal moth (IMM, Plodia interpunctella) colony exhibiting resistance to Bt. We found a number of changes in the levels of certain specific midgut proteins that indicate increased glutathione utilization, elevation in oxidative metabolism, and differential maintenance of energy balance within the midgut epithelial cells of the Bt-resistant IMM larva. Additionally, the electrophoretic migration pattern of a low molecular mass acidic protein, which apparently is an ortholog of F(1)F(0)-ATPase, was considerably altered in the Bt-resistant insect indicating that variations in amino acid content or modifications of certain proteins also are important components of the resistance phenomenon in the IMM. Furthermore, there was a dramatic decrease in the level of chymotrypsin-like proteinase in the midgut of the Bt-resistant larva, signifying that reduction of chymotrypsin activity, and subsequently decreased activation of Cry toxin in the insect midgut, is an important factor in the resistant state of the IMM. The proteomic analysis of larval gut proteins utilized in this study provides a useful approach for consolidating protein changes and physiological events associated with insect resistance to Bt. Our results support the hypothesis that physiological adaptation of insects and resistance to Bt is multifaceted, including protein modification and changes in the synthesis of specific larval gut proteins. We believe that increased oxidative metabolism may be an adaptive response of insects that undergo survival challenge and that it could mediate detoxification as well as higher rates of generalized and localized mutations that enhance their resistance and provide survival advantage.

Animals↗
PubMed2004

Monocyte chemoattractant protein-1 in patients with peripheral arterial disease.

BACKGROUND: Chemokine-driven migration of inflammatory cells has been implicated in the pathogenesis of atherosclerotic conditions including peripheral arterial disease (PAD). Monocyte chemoattractant protein-1 (MCP-1) is elevated in patients with coronary artery disease and in hypertensive patients. This study therefore investigated MCP-1 in patients with PAD. METHODS: Serum MCP-1 was determined by enzyme-linked immunosorbent assay in 36 healthy, control subjects and in 19 patients with PAD. Statistical analysis utilised the Mann-Whitney test and Spearman correlation (p < 0.05). RESULTS: MCP-1 (pg/ml) was increased in patients compared with in controls (mean+/-standard error of the mean: PAD group, 748+/-60; control group, 459+/-27; p=0.0001). MCP-1 levels tended to decrease with progressing disease. From atherosclerosis risk factors, diabetes inclined to increase MCP-1 levels; hypertension had no effect. Serum MCP-1 correlated with cholesterol, triglycerides, low-density lipoprotein but not high-density lipoprotein. CONCLUSION: Elevation of MCP-1 in the circulation of PAD patients shown in the present pilot study implicates this CC chemokine ligand 2 in inflammatory processes contributing to PAD clinical symptomatology. Further investigations are necessary to evaluate whether MCP-1 can be used as a potential marker of peripheral arterial disease follow-up and/or prognosis.

Aged↗
PubMed1990

Ground distribution patterns of selected radioactive, chemical, and physical contaminants from dispersion of U mill tailings.

Ground concentrations of 226Ra and total alpha activity as a function of soil depth and distance from an acidic U mill tailings pile were measured, as well as soil pH, sulfate, chloride, and conductivity. Contamination decreased with increasing soil depth and distance from the source. Most of the measured parameters were indistinguishable from background after a distance of 3.3 km from the tailings impoundment. The highest concentrations of all contaminants were contained in the particles less than 0.045 mm in diameter at the soil surface (0-0.6 cm). This fraction represents only 4% to 10% of the total soil mass. Surface soil particles greater than 2.0 mm contained higher concentrations of all contaminants than the intermediate-size (0.045-2.0 mm) fraction. Aggregate formation of the greater than 2.0-mm fraction from small particles or preferential erosion and transport of the tailings sand fractions were proposed as possible explanations. The significant correlations between various measurements were soil 226Ra vs. total alpha activity, sulfate concentration, and conductivity. The latter two parameters are indicative of sulfuric acid leached tailings. The sulfate concentration appeared to be a sensitive and easily measured indicator of tailings contamination and migration into the soil profile. Furthermore, sulfate may be used to delineate acidic U tailings from other sources of contamination (U ore dust, yellowcake, naturally elevated U and Ra) in the environment. Actual site measurements for surface 226Ra concentration were approximately twice the MILDOS computer code prediction.

Chlorides↗
PubMedDate not supplied

FAK signaling in anaplastic astrocytoma and glioblastoma tumors.

Focal adhesion kinase (FAK) is a non-receptor cytoplasmic-tyrosine kinase that is activated by several different cell surface receptors shown to be upregulated on glioblastoma cells (integrins alpha(v)beta3 and alpha(v)beta5, and the epidermal growth factor receptor). Activated FAK can signal through several different signaling pathways, which are reviewed here. Published data are summarized that have demonstrated 1) elevated FAK expression in anaplastic astrocytoma and glioblastoma tumor biopsy samples, 2) a role for FAK in the promotion of glioblastoma cell proliferation, survival and migration in vitro, and 3) a role for FAK in the promotion of glioblastoma cell proliferation in vivo in an animal model. The available data suggests that increased levels of FAK protein and activity may contribute to an increased ERK activity and cell proliferation in vivo in these tumors.

Astrocytoma↗
PubMed2004

Role of anti-CD3 in modulation of Th1-type immune response in Shigella dysenteriae infection.

A murine model was used to evaluate the role of anti-CD3 in modulating a Th1-type response by restimulation of T-cells after immunization with the 57 kDa immunodominant antigen of Shigella dysenteriae 1 outer-membrane proteins (OMPs), followed by Shigella infection after immunization. To observe the effect of anti-CD3, other T-cell cultures were also established following anti-CD1, anti-IL2 and phytohaemagglutinin stimulation. Anti-CD3 stimulation of reconstituted T-cells showed 'mean' levels of CD4 and CD25 were enhanced by 34.5 and 31.1 % in immunized mice, which was comparable to 53.2 and 50.7 %, respectively, in challenged-immunized mice, and were dominant over CD8+ T-cells. Levels of IL2 generated by anti-CD3-stimulated T-cells of immunized mice were greater than those of unstimulated T-cells and were significantly elevated in challenged-immunized mice. The reactivity of T-cells indicated their complete responsiveness, as anti-CD3 antibody might not inhibit the migration of the macrophages but rather inhibit IL4. These macrophage factors synergistically act with anions towards an activated response, which in turn provokes IL2 secretion with a low degree of internalization of its receptor. Thus, sharing of IL2 to form a high-affinity receptor complex with CD4+ T-cells through motive signals suggested a generalized T-cell activation with increased humoral responses. Macrophage migration inhibition factor (MIF) and IL4 responses during anti-CD3 stimulation of immunized mice indicated that the role of anti-CD3 in generation of O2- is due to a synergistic effect by Th1 subsets of Th0 cells. The above findings should have implications for understanding the immunoregulatory role of anti-CD3 associated with 57 kDa antigen in immunoprophylactic measures.

Animals↗