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Human Trafficking

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3281. Dynamics of platelet-activating-factor release and uptake in a human neutrophil suspension. Full Text available with Trip Pro

Dynamics of platelet-activating-factor release and uptake in a human neutrophil suspension. The present study has examined the dynamics of platelet-activating-factor (PAF) synthesis, release and uptake in order to understand better the trafficking of PAF between cells and medium. Initial experiments indicated that the amount of PAF found on the outside of the cell remained constant well after the synthesis from a precursor had apparently stopped, and in spite of a continued capacity of the cell (...) . The potential for PAF and/or its intermediates to be recycled was verified by demonstrating that a large proportion of exogenously provided 1-alkyl-2-lyso-sn-glycero-3-phosphocholine is taken up by the neutrophil, converted into PAF and then released again by the cell. These results suggest that PAF trafficking between the cell and medium is complex and involves many processes, which include synthesis, release, uptake, catabolism and recycling.

1991 Biochemical Journal

3282. Human Papillomavirus Types 16, 31, and 58 Use Different Endocytosis Pathways To Enter Cells Full Text available with Trip Pro

Human Papillomavirus Types 16, 31, and 58 Use Different Endocytosis Pathways To Enter Cells The early steps of the intracellular trafficking of human papillomavirus type 16 (HPV-16), -31, and -58 pseudovirions were studied by investigating the effects of drugs acting at defined points of endocytosis pathways on virus-like particle-mediated pseudoinfection by overexpression of a dominant-negative form of the Eps15 protein to inhibit clathrin-mediated endocytosis and by electron microscopy

2003 Journal of virology

3283. Identification and subcellular localization of human rab5b, a new member of the ras-related superfamily of GTPases. Full Text available with Trip Pro

Identification and subcellular localization of human rab5b, a new member of the ras-related superfamily of GTPases. Members of the mammalian rab family of GTPases are associated with specific subcellular compartments, where these proteins are postulated to function in vesicular transport. By screening a human umbilical vein endothelial cell library with degenerate oligonucleotide probes, we have isolated a 1.6-kb cDNA clone encoding a 215-amino-acid protein belonging to the rab family (...) of GTPases. This newly identified rab protein is 81% identical to human rab5, the canine counterpart of which has been localized to the plasma membrane and early endosomes. In light of this homology, we have named this new member of the GTPase superfamily "rab5b." Northern analysis using the rab5b cDNA as a probe revealed a 3.6-kb mRNA in a variety of cell types, including human umbilical vein endothelial cells, K562 erythroleukemia cells, U937 monoblastic cells, and HeLa cells. A fusion protein between

1992 Journal of Clinical Investigation

3284. Modification of host cell membrane lipid composition by the intra-erythrocytic human malaria parasite Plasmodium falciparum. Full Text available with Trip Pro

Modification of host cell membrane lipid composition by the intra-erythrocytic human malaria parasite Plasmodium falciparum. The phospholipid and fatty acid compositions of the host infected erythrocyte plasma membrane (IEPM) have been determined for erythrocytes infected with the human malaria parasite Plasmodium falciparum. IEPM were prepared by selective lysis of the host erythrocyte (but not of the parasite membranes) with 0.1% saponin, followed by differential centrifugation. The purity (...) of the IEPM was determined by measuring the membrane-specific enzyme markers acetylcholinesterase, glutamate dehydrogenase and lactate dehydrogenase, and by immunoelectron microscopy using monoclonal antibodies specific for human erythrocyte glycophorin A (4E7) and for a 195 kDa parasite membrane glycoprotein (Pf6 3B10.1). Both approaches demonstrated that the host erythrocyte plasma membrane preparation was free from contamination by parasite membranes. During intra-erythrocytic development

1991 Biochemical Journal

3285. B lymphocyte binding to E- and P-selectins is mediated through the de novo expression of carbohydrates on in vitro and in vivo activated human B cells. Full Text available with Trip Pro

B lymphocyte binding to E- and P-selectins is mediated through the de novo expression of carbohydrates on in vitro and in vivo activated human B cells. Cell adhesion to endothelium regulates the trafficking and recruitment of leukocytes towards lymphoid organs and sites of inflammation. This phenomenon is mediated by the expression of a number of adhesion molecules on both the endothelium and circulating cells. Activation of endothelial cells (EC) with different stimuli induces the expression

1994 Journal of Clinical Investigation

3286. Metabolic characteristics of a human hepatoma cell line stably transfected with hormone-sensitive lipase. Full Text available with Trip Pro

Metabolic characteristics of a human hepatoma cell line stably transfected with hormone-sensitive lipase. Clones of HepG2 cells were selected that stably express the cDNA for hormone-sensitive lipase (HSL). When cells were cultured in the presence of labelled extracellular oleate, accumulation of labelled fatty acid as cellular triacylglycerol (TAG) was significantly lower in the transfectants compared with the wild-type cells. There was no change in the net rate of phospholipid (PL) synthesis (...) . Culture of cells containing isotopically prelabelled TAG resulted in a greater net loss of TAG from the transfected cells than from the wild-type cells. The excess loss of labelled TAG was primarily due to an increased TAG fatty acid oxidation. Free fatty acid release into the medium was not increased in the transfectants, nor was the very low rate of lipoprotein lipid secretion. Also, there was no increased net trafficking of fatty acids from TAG into PLs. Changes in the 3H:14C ratio of TAG

1999 Biochemical Journal

3287. A human homologue of Escherichia coli ClpP caseinolytic protease: recombinant expression, intracellular processing and subcellular localization. Full Text available with Trip Pro

A human homologue of Escherichia coli ClpP caseinolytic protease: recombinant expression, intracellular processing and subcellular localization. We have recently cloned a human cDNA (hClpP) with significant sequence similarity to the ATP-dependent Escherichia coli ClpP protease [Bross, Andresen, Knudsen, Kruse and Gregersen (1995) FEBS Lett. 377, 249-252]. In the present study, synthesis, intracellular processing and subcellular localization of hClpP have been analysed in intact cells (...) of fluorogenic peptides could be observed in extracts of Chang cells overexpressing hClpP, indicating that the protease may not be active without co-factors. Immunofluorescence studies using confocal-laser-scanning microscopy showed co-localization of the hClpP and the mitochondrially located Hsp60 (heat-shock protein 60). Taken together, the results reported here show that hClpP is localized inside mitochondria and that the trafficking and processing of hClpP resembles the typical biogenesis pathway

1998 Biochemical Journal

3288. MCP-1 deficiency reduces susceptibility to atherosclerosis in mice that overexpress human apolipoprotein B Full Text available with Trip Pro

MCP-1 deficiency reduces susceptibility to atherosclerosis in mice that overexpress human apolipoprotein B The earliest recognizable atherosclerotic lesions are fatty streaks composed of lipid-laden macrophages (foam cells). Circulating monocytes are the precursors of these foam cells, but the molecular mechanisms that govern macrophage trafficking through the vessel wall are poorly understood. Monocyte chemoattractant protein-1 (MCP-1), a member of the chemokine (chemotactic cytokine) family (...) , is a potent monocyte agonist that is upregulated by oxidized lipids. Recent studies in hypercholesterolemic mice lacking apo E or the low-density lipoprotein receptor have suggested a role for MCP-1 in monocyte recruitment to early atherosclerotic lesions. To determine if MCP-1 is critically involved in atherogenesis in the setting of elevated physiological plasma cholesterol levels, we deleted the MCP-1 gene in transgenic mice expressing human apo B. Here we report that the absence of MCP-1 provides

1999 Journal of Clinical Investigation

3289. Transcriptional programs activated by exposure of human prostate cancer cells to androgen Full Text available with Trip Pro

Transcriptional programs activated by exposure of human prostate cancer cells to androgen Androgens are required for both normal prostate development and prostate carcinogenesis. We used DNA microarrays, representing approximately 18,000 genes, to examine the temporal program of gene expression following treatment of the human prostate cancer cell line LNCaP with a synthetic androgen.We observed statistically significant changes in levels of transcripts of more than 500 genes. Many (...) of these genes were previously reported androgen targets, but most were not previously known to be regulated by androgens. The androgen-induced expression programs in three additional androgen-responsive human prostate cancer cell lines, and in four androgen-independent subclones derived from LNCaP, shared many features with those observed in LNCaP, but some differences were observed. A remarkable fraction of the genes induced by androgen appeared to be related to production of seminal fluid and these genes

2002 Genome Biology

3290. In Vivo Regulation of Human Skeletal Muscle Gene Expression by Thyroid Hormone Full Text available with Trip Pro

In Vivo Regulation of Human Skeletal Muscle Gene Expression by Thyroid Hormone Thyroid hormones are key regulators of metabolism that modulate transcription via nuclear receptors. Hyperthyroidism is associated with increased metabolic rate, protein breakdown, and weight loss. Although the molecular actions of thyroid hormones have been studied thoroughly, their pleiotropic effects are mediated by complex changes in expression of an unknown number of target genes. Here, we measured patterns (...) of skeletal muscle gene expression in five healthy men treated for 14 days with 75 microg of triiodothyronine, using 24,000 cDNA element microarrays. To analyze the data, we used a new statistical method that identifies significant changes in expression and estimates the false discovery rate. The 381 up-regulated genes were involved in a wide range of cellular functions including transcriptional control, mRNA maturation, protein turnover, signal transduction, cellular trafficking, and energy metabolism

2002 Genome Research

3291. Effect of an acute oral lithium intake on urinary Aquaporin-2 in healthy humans with and without simultaneous stimulation with hypertonic saline infusion. (Abstract)

lithium was given. It is suggested that lithium increases urinary output by inhibiting trafficking of renal water channels in healthy humans. (...) Effect of an acute oral lithium intake on urinary Aquaporin-2 in healthy humans with and without simultaneous stimulation with hypertonic saline infusion. Animal experiments have shown that lithium interferes with the formation of Aquaporin-2 in the distal renal tubuli. The effect of lithium on formation of renal water channels has not been studied in healthy humans. The aim of this study was to test the hypotheses that a single oral dose of lithium will reduce the formation of water channels

2003 Scandinavian journal of clinical and laboratory investigation Controlled trial quality: uncertain

3292. Long-term in vivo survival of receptor-modified syngeneic T cells in patients with human immunodeficiency virus infection. (Abstract)

Long-term in vivo survival of receptor-modified syngeneic T cells in patients with human immunodeficiency virus infection. To study human immunodeficiency virus (HIV)-specific cellular immunity in vivo, we transferred syngeneic lymphocytes after ex vivo expansion and transduction with a chimeric receptor gene (CD4/CD3-zeta) between identical twins discordant for HIV infection. Single and multiple infusions of 10(10) genetically modified CD8(+) T cells resulted in peak fractions (...) of approximately 10(3) to 10(4) modified cells/10(6) total CD4(+) or CD8(+) cells persisted for at least 1 year. Assessment of in vivo trafficking of the transferred cells by lymphoid tissue biopsies revealed the presence of modified cells in proportions equivalent to or below those in the circulation. The cell infusions were well tolerated and were not associated with substantive immunologic or virologic changes. Thus, adoptive transfer of genetically modified HIV-antigen-specific T cells was safe. Sustained

2000 Blood

3293. A randomized placebo-controlled trial of a humanized monoclonal antibody to alpha4 integrin in active Crohn's disease. (Abstract)

A randomized placebo-controlled trial of a humanized monoclonal antibody to alpha4 integrin in active Crohn's disease. alpha4 integrins are important mediators of leukocyte migration across vascular endothelium. This pilot placebo-controlled study aimed to assess the safety and efficacy of natalizumab, a recombinant humanized monoclonal antibody to alpha4 integrin, in patients with mild to moderately active Crohn's disease.Thirty patients with active Crohn's disease (Crohn's Disease Activity (...) lymphocyte trafficking. Natalizumab therapy in active Crohn's disease merits further investigation.

2001 Gastroenterology Controlled trial quality: predicted high

3294. Human anion exchanger1 mutations and distal renal tubular acidosis. (Abstract)

Human anion exchanger1 mutations and distal renal tubular acidosis. The human anion exchanger 1 (AE1 or SLC4A1) gene encodes anion exchanger 1 (or band 3) protein in erythrocytes and in alpha-intercalated cells of the kidney. Thus, AE1 mutations show pleiotrophic effects resulting in two distinct and seemingly unrelated defects, an erythrocyte abnormality and distal renal tubular acidosis (dRTA). Southeast Asian ovalocytosis (SAO), a well-known red blood cell (RBC) defect, which is widespread (...) . Therefore, AE1 mutations can result in both recessive and dominant dRTA, possibly depending on the position of the amino acid change in the protein. As several mutant AE1 proteins still maintain a significant anion transport function but are defective in targeting to the cell surface, impaired intracellular trafficking of the mutant AE1 is an important molecular mechanism involved in the pathogenesis of dRTA associated with AE1 mutations.

2003 Southeast Asian Journal of Tropical Medicine and Public Health

3295. Caveolin expression and localization in human keratinocytes suggest a role in lamellar granule biogenesis. Full Text available with Trip Pro

as caveolae. Similarities in the composition of lamellar granules and caveolae suggest that caveolins could be involved in lamellar granule assembly, trafficking, and/or function. In order to explore this relationship, we have examined the expression of caveolins in epidermis, keratinocyte cultures, and an isolated lamellar granule fraction using immunolabeling, immunoblotting, and northern blotting. Several antibodies show immunolocalization of caveolin-1 in the basal layer of human epidermis (...) Caveolin expression and localization in human keratinocytes suggest a role in lamellar granule biogenesis. Lamellar granules are sphingolipid-enriched organelles, probably intimately related to the tubulo-vesicular elements of the trans-Golgi network, that deliver the precursors of stratum corneum barrier lipids to the extracellular compartment. Caveolins are cholesterol-binding scaffolding proteins that facilitate the assembly of cholesterol- and sphingolipid-enriched membrane domains known

2003 Journal of Investigative Dermatology

3296. Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult. Full Text available with Trip Pro

Spectrum and range of oxidative stress responses of human lens epithelial cells to H2O2 insult. Oxidative stress (OS) is believed to be a major contributor to age-related cataract and other age-related diseases.cDNA microarrays were used to identify the spectrum and range of genes with transcript levels that are altered in response to acute H(2)O(2)-induced OS in human lens epithelial (HLE) cells. HLE cells were treated with 50 microM H(2)O(2) for 1 hour in the absence of serum, followed (...) by a return to complete medium. RNAs were prepared from treated and untreated cells at 0, 1, 2, and 8 hours after H(2)O(2) treatment.The data showed 1171 genes that were significantly up- and downregulated in response to H(2)O(2) treatment. Several functional subcategories of genes were identified, including those encoding DNA repair proteins, antioxidant defense enzymes, molecular chaperones, protein biosynthesis enzymes, and trafficking and degradation proteins. Differential expression of selected genes

2003 Investigative Ophthalmology & Visual Science

3297. Water permeability of C-terminally truncated aquaporin 0 (AQP0 1-243) observed in the aging human lens. (Abstract)

Water permeability of C-terminally truncated aquaporin 0 (AQP0 1-243) observed in the aging human lens. To first assess the distribution of posttranslationally truncated products of aquaporin 0 (AQP0) in dissected sections of a normal human lens and to determine the effect of backbone cleavage on the water permeability of AQP0.A 27-year-old lens was concentrically dissected into six sections. Membrane protein was isolated from each section and cleaved with cyanogen bromide, and the peptides (...) , and 243 on AQP0 water permeability were examined. Truncation after residue 243 resulted in an approximate 15% decrease in permeability compared with the full-length protein, AQP0 1-263. However, rather than a direct effect on water transport, analysis of surface protein expression indicated that the decrease in permeability was a result of less efficient protein trafficking to the oocyte surface and that the permeabilities of full-length and 1-243 AQP0 were indistinguishable. Further, C-terminal

2003 Investigative Ophthalmology & Visual Science

3298. Mutations associated with neutropenia in dogs and humans disrupt intracellular transport of neutrophil elastase. (Abstract)

hematopoiesis. C-terminal processing of neutrophil elastase exposes an AP3 interaction signal responsible for redirecting neutrophil elastase trafficking from membranes to granules. Disruption of either neutrophil elastase or AP3 perturbs the intracellular trafficking of neutrophil elastase. Most mutations in ELA2 that cause human cyclic hematopoiesis prevent membrane localization of neutrophil elastase, whereas most mutations in ELA2 that cause SCN lead to exclusive membrane localization. (...) Mutations associated with neutropenia in dogs and humans disrupt intracellular transport of neutrophil elastase. Cyclic hematopoiesis is a stem cell disease in which the number of neutrophils and other blood cells oscillates in weekly phases. Autosomal dominant mutations of ELA2, encoding the protease neutrophil elastase, found in lysosome-like granules, cause cyclic hematopoiesis and most cases of the pre-leukemic disorder severe congenital neutropenia (SCN; ref. 3) in humans. Over 20

2003 Nature Genetics

3299. Human hepatic stellate cells show features of antigen-presenting cells and stimulate lymphocyte proliferation. (Abstract)

Human hepatic stellate cells show features of antigen-presenting cells and stimulate lymphocyte proliferation. Following cell activation, hepatic stellate cells (HSCs) acquire proinflammatory and profibrogenic properties. We investigated whether activated HSCs also display immune properties. Here we show that cultured human HSCs express membrane proteins involved in antigen presentation, including members of the HLA family (HLA-I and HLA-II), lipid-presenting molecules (CD1b and CD1c (...) ), and factors involved in T-cell activation (CD40 and CD80). Exposure of HSCs to proinflammatory cytokines markedly up-regulates these molecules. Importantly, cells freshly isolated from human cirrhotic livers (in vivo activated HSCs) highly express HLA-II and CD40, suggesting that HSCs can act as antigen-presenting cells (APCs) in human fibrogenesis. We also explored whether human HSCs can efficiently process exogenous antigens. Activated HSCs internalize low- and high-molecular-weight dextran

2003 Hepatology

3300. Functional expression of the human thiazide-sensitive NaCl cotransporter in Madin-Darby canine kidney cells. (Abstract)

Functional expression of the human thiazide-sensitive NaCl cotransporter in Madin-Darby canine kidney cells. The thiazide-sensitive Na(+)-Cl(-) cotransporter (NCC), which is expressed on the apical membrane of epithelial cells lining the distal convoluted tubule, is responsible for the reabsorption of 5% to 10% of filtered Na(+) and Cl(-). To date, functional studies on the structural and regulatory requirements for localized trafficking and ion-transporting activity of NCC have been hampered (...) by lack of a suitable cell system expressing this cotransporter. Reported here is the functional expression of human NCC (hNCC) in a polarized mammalian cell of renal origin-that is, the high-resistance Madin-Darby canine kidney (MDCK) cell. Western blot testing revealed that the cells predominantly expressed the complex glycosylated (approximately 140 kD) form of hNCC. hNCC was present primarily in the apical part of the cell. The functionality of hNCC was demonstrated by the gain of thiazide

2003 Journal of the American Society of Nephrology

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