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1.
Med Sci Law ; 63(1): 53-60, 2023 Jan.
Article in English | MEDLINE | ID: mdl-35934979

ABSTRACT

Objective: Opioid substitution treatment (OST) is a common treatment for individuals who use opioids; however, empirical evidence on the effects of OST during incarceration is scarce. Our aim was to conduct a meta-analysis on the effects of incarceration-based OST on substance use, treatment engagement post-release and re-incarceration. Method: We searched for studies on individuals who were incarcerated and treated with OST, compared to a comparison group. Studies were only included if they reported data post-release. Results: N = 15 studies met the inclusion criteria. We found less opioid use, less other drug use, higher treatment engagement post-release and less re-incarceration among treated individuals compared to the comparison group. Moderator analyses showed some influence of length of follow-up period and study quality. Conclusions: Incarceration-based OST reduces drug use, re-incarceration and leads to higher treatment engagement after release. More research is needed on the effects of incarceration-based OST on secondary outcomes (e.g. health and social integration) and on factors that moderate these effects.


Subject(s)
Opioid-Related Disorders , Prisoners , Humans , Opiate Substitution Treatment , Opioid-Related Disorders/drug therapy , Analgesics, Opioid/therapeutic use
2.
J Thromb Haemost ; 14(11): 2212-2226, 2016 11.
Article in English | MEDLINE | ID: mdl-27590316

ABSTRACT

Essentials The role of protein C (PC) activation in experimental autoimmune encephalitis (EAE) is unknown. PC activation is required for mitochondrial function in the central nervous system. Impaired PC activation aggravates EAE, which can be compensated for by soluble thrombomodulin. Protection of myelin by activated PC or solulin is partially independent of immune-modulation. SUMMARY: Background Studies with human samples and in rodents established a function of coagulation proteases in neuro-inflammatory demyelinating diseases (e.g. in multiple sclerosis [MS] and experimental autoimmune encephalitis [EAE]). Surprisingly, approaches to increase activated protein C (aPC) plasma levels as well as antibody-mediated inhibition of PC/aPC ameliorated EAE in mice. Hence, the role of aPC generation in demyelinating diseases and potential mechanisms involved remain controversial. Furthermore, it is not known whether loss of aPC has pathological consequences at baseline (e.g. in the absence of disease). Objective To explore the role of thrombomodulin (TM)-dependent aPC generation at baseline and in immunological and non-immunological demyelinating disease models. Methods Myelination and reactive oxygen species (ROS) generation were evaluated in mice with genetically reduced TM-mediated protein C activation (TMPro/Pro ) and in wild-type (WT) mice under control conditions or following induction of EAE. Non-immunological demyelination was analyzed in the cuprizone-diet model. Results Impaired TM-dependent aPC generation already disturbs myelination and mitochondrial function at baseline. This basal phenotype is linked with increased mitochondrial ROS and aggravates EAE. Reducing mitochondrial ROS (p66Shc deficiency), restoring aPC plasma levels or injecting soluble TM (solulin) ameliorates EAE in TMPro/Pro mice. Soluble TM additionally conveyed protection in WT-EAE mice. Furthermore, soluble TM dampened demyelination in the cuprizone-diet model, demonstrating that its myelin-protective effect is partially independent of an immune-driven process. Conclusion These results uncover a novel physiological function of TM-dependent aPC generation within the CNS. Loss of TM-dependent aPC generation causes a neurological defect in healthy mice and aggravates EAE, which can be therapeutically corrected.


Subject(s)
Central Nervous System/metabolism , Mitochondria/metabolism , Myelin Sheath/chemistry , Protein C/metabolism , Thrombomodulin/blood , Animals , Brain/metabolism , Cardiolipins/chemistry , Encephalomyelitis, Autoimmune, Experimental/immunology , Encephalomyelitis, Autoimmune, Experimental/metabolism , Humans , Immune System , Mice , Mice, Inbred C57BL , Mutation , Neurons , Oxidative Stress , PC12 Cells , Phenotype , Rats , Reactive Oxygen Species/metabolism , Solubility , Thrombomodulin/chemistry
3.
Cell Death Dis ; 1: e58, 2010 Jul 22.
Article in English | MEDLINE | ID: mdl-21364663

ABSTRACT

Oxidative stress leads to T-cell hyporesponsiveness or death. The actin-binding protein cofilin is oxidized during oxidative stress, which provokes a stiff actin cytoskeleton and T-cell hyporesponsiveness. Here, we show that long-term oxidative stress leads to translocation of cofilin into the mitochondria and necrotic-like programmed cell death (PCD) in human T cells. Notably, cofilin mutants that functionally mimic oxidation by a single mutation at oxidation-sensitive cysteins (Cys-39 or Cys-80) predominately localize within the mitochondria. The expression of these mutants alone ultimately leads to necrotic-like PCD in T cells. Accordingly, cofilin knockdown partially protects T cells from the fatal effects of long-term oxidative stress. Thus, we introduce the oxidation and mitochondrial localization of cofilin as the checkpoint for necrotic-like PCD upon oxidative stress as it occurs, for example, in tumor environments.


Subject(s)
Caspases/metabolism , Cofilin 1/metabolism , Mitochondria/metabolism , Necrosis/metabolism , Oxidative Stress , T-Lymphocytes/metabolism , Apoptosis , Cells, Cultured , Cofilin 1/genetics , HSP70 Heat-Shock Proteins/metabolism , Humans , Hydrogen Peroxide/pharmacology , Oxidation-Reduction , RNA Interference , RNA, Small Interfering/metabolism , T-Lymphocytes/immunology
4.
Chirurg ; 77(9): 844-55, 2006 Sep.
Article in German | MEDLINE | ID: mdl-16821051

ABSTRACT

BACKGROUND: Current German legislation ( (section sign) 115 b SGB V) allows groin hernia inpatient treatment only under particular circumstances. That allows the operative technique of first choice for outpatient groin hernia repair to be determined by basic market principles. The aim of this paper was to study the feasibility of outpatient minimally invasive hernia surgery with regard to complication rates, patient satisfaction, and economic considerations. METHODS: For 1 year, a total of 571 patients with inguinal hernias (131 male, eight female, mean age 46 years, all ASA I) were treated at two surgical centers. Twenty-four percent (139/571) underwent outpatient total extraperitoneal repair (TEP). Complication rates were recorded. Patient satisfaction with the procedure was evaluated by a standard questionnaire. Cost calculations were compared with revenues according to the EBM2000plus. RESULTS: Of the patients, 96.4% were discharged on the day of operation without subsequent rehospitalization, 84% had no fears of complications at home, 54% went back to work in less than 14 days, and 88.7% were willing to undergo TEP a second time if necessary. Calculated average total cost of euro 709 exceeded the revenue of euro 565 by 20%. CONCLUSION: For a carefully selected group, outpatient TEP is patient-friendly and safe. Despite these advantages, it still remains economically unattractive to hospital management because of the 20% cover shortage. Improvements in the current legislation are urgently desired.


Subject(s)
Ambulatory Surgical Procedures/legislation & jurisprudence , Hernia, Inguinal/surgery , Minimally Invasive Surgical Procedures/legislation & jurisprudence , National Health Programs/legislation & jurisprudence , Outcome Assessment, Health Care/legislation & jurisprudence , Adult , Ambulatory Surgical Procedures/economics , Ambulatory Surgical Procedures/statistics & numerical data , Costs and Cost Analysis , Female , Germany , Health Care Costs/legislation & jurisprudence , Health Care Costs/statistics & numerical data , Hernia, Inguinal/economics , Hernia, Inguinal/epidemiology , Humans , Male , Middle Aged , Minimally Invasive Surgical Procedures/economics , Minimally Invasive Surgical Procedures/statistics & numerical data , National Health Programs/economics , National Health Programs/statistics & numerical data , Outcome Assessment, Health Care/economics , Outcome Assessment, Health Care/statistics & numerical data , Patient Admission/economics , Patient Admission/legislation & jurisprudence , Patient Admission/statistics & numerical data , Patient Satisfaction , Postoperative Complications/economics , Postoperative Complications/epidemiology , Reoperation
5.
J Mol Biol ; 311(4): 709-21, 2001 Aug 24.
Article in English | MEDLINE | ID: mdl-11518525

ABSTRACT

SNARE proteins are essential for different types of intracellular membrane fusion. Whereas interaction between their cytoplasmic domains is held responsible for establishing membrane proximity, the role of the transmembrane segments in the fusion process is currently not clear. Here, we used an in vitro approach based on lipid mixing and electron microscopy to examine a potential fusogenic activity of the transmembrane segments. We show that the presence of synthetic peptides representing the transmembrane segments of the presynaptic soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) synaptobrevin II (also referred to as VAMP II) or syntaxin 1A, but not of an unrelated control peptide, in liposomal membranes drives their fusion. Liposome aggregation by millimolar Ca(2+) concentrations strongly potentiated the effect of the peptides; this indicates that juxtaposition of the bilayers favours their fusion in the absence of the cytoplasmic SNARE domains. Peptide-driven fusion is reminiscent of natural membrane fusion, since it was suppressed by lysolipid and involved both bilayer leaflets. This suggests transient presence of a hemifusion intermediate followed by complete membrane merger. Structural studies of the peptides in lipid bilayers performed by Fourier transform infrared spectroscopy indicated mixtures of alpha-helical and beta-sheet conformations. In isotropic solution, circular dichroism spectroscopy showed the peptides to exist in a concentration-dependent equilibrium of alpha-helical and beta-sheet structures. Interestingly, the fusogenic activity decreased with increasing stability of the alpha-helical solution structure for a panel of variant peptides. Thus, structural plasticity of transmembrane segments may be important for SNARE protein function at a late step in membrane fusion.


Subject(s)
Membrane Fusion , Membrane Proteins/chemistry , Membrane Proteins/metabolism , Molecular Mimicry , Peptides/chemistry , Peptides/metabolism , Vesicular Transport Proteins , Amino Acid Sequence , Calcium/metabolism , Circular Dichroism , Lipid Bilayers/chemistry , Lipid Bilayers/metabolism , Liposomes/chemistry , Liposomes/metabolism , Microscopy, Electron , Molecular Sequence Data , Pliability , Protein Structure, Secondary , Protein Structure, Tertiary , Qa-SNARE Proteins , R-SNARE Proteins , SNARE Proteins , Solutions , Spectroscopy, Fourier Transform Infrared
6.
Eur J Immunol ; 31(6): 1813-24, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11433378

ABSTRACT

The role of astrocytes and microglia as antigen-presenting cells in the brain is still controversial. In this study we have analyzed and compared aspects of the molecular machinery that underlies MHC class II trafficking in immunocompetent astrocytes and microglia. We show that IFN-gamma-stimulated microglia possess active cathepsin L and cathepsin S, and efficiently degrade the invariant chain, unlike IFN-gamma-stimulated astrocytes that express cathepsin L but not cathepsin S. The lack of cathepsin S proves to be dramatic for the antigen-presentation capacity of astrocytes, which is nearly abolished when these cells are stimulated by a combination of IFN-gamma and TNF-alpha. TNF-alpha indeed decreases cathepsin L activity as we show here, leading to alterations in invariant chain processing, and hence in MHC class II trafficking in astrocytes. Cystatin C inhibits cathepsin L activity in astrocytes, but does not regulate cathepsin L and cathepsin S activity in microglia. We therefore identify cathepsin L and cathepsin S as key components in the regulation of the immune potential of astrocytes and microglia, and provide evidence for a cell-specific regulation exerted by IFN-gamma and TNF-alpha on the expression and activity of cathepsins.


Subject(s)
Antigen Presentation/immunology , Antigens, Differentiation, B-Lymphocyte/immunology , Astrocytes/immunology , Cathepsins/immunology , Endopeptidases , Histocompatibility Antigens Class II/immunology , Microglia/immunology , Antigen Presentation/drug effects , Astrocytes/cytology , Astrocytes/drug effects , Cathepsin L , Cells, Cultured , Cysteine Endopeptidases , Endosomes/ultrastructure , Humans , Immunocompetence , Interferon-gamma/immunology , Interferon-gamma/pharmacology , Lysosomes/ultrastructure , Microglia/cytology , Microglia/drug effects , Tumor Necrosis Factor-alpha/immunology , Tumor Necrosis Factor-alpha/pharmacology
7.
Biochemistry ; 39(43): 13166-75, 2000 Oct 31.
Article in English | MEDLINE | ID: mdl-11052669

ABSTRACT

Abnormal tau-immunoreactive filaments are a hallmark of tauopathies, including Alzheimer's disease (AD). A higher phosphorylation ("hyperphosphorylation") state of tau protein may represent a critical event. To determine the potential role of tau hyperphosphorylation in these disorders, mutated tau proteins were produced where serine/threonine residues known to be highly phosphorylated in tau filaments isolated from AD patients were substituted for glutamate to simulate a paired helical filament (PHF)-like tau hyperphosphorylation. We demonstrate that, like hyperphosphorylation, glutamate substitutions induce compact structure elements and SDS-resistant conformational domains in tau protein. Hyperphosphorylation-mimicking glutamate-mutated tau proteins display a complete functional loss in its ability to promote microtubule nucleation which can partially be overcome by addition of the osmolyte trimethylamine N-oxide (TMAO), which is similar to phosphorylated tau. In addition, glutamate-mutated tau proteins fail to interact with the dominant brain protein phosphatase 2A isoform ABalphaC, and exhibit a reduced ability to assemble into filaments. Interestingly, wild-type tau and phosphorylation-mimicking tau similarly bind to microtubules when added alone, but the mutated tau is almost completely displaced from the microtubule surface by equimolar concentrations of wild-type tau. The data indicate that glutamate-mutated tau proteins provide a useful model for analyzing the functional consequences of tau hyperphosphorylation. They suggest that several mechanisms contribute to the abnormal tau accumulation observed during tauopathies, in particular a selective displacement of hyperphosphorylated tau from microtubules, a functional loss in promoting microtubule nucleation, and a failure to interact with phosphatases.


Subject(s)
Molecular Mimicry/genetics , Mutagenesis, Site-Directed , tau Proteins/genetics , tau Proteins/metabolism , Actin Cytoskeleton/genetics , Actin Cytoskeleton/metabolism , Actin Cytoskeleton/pathology , Adult , Amino Acid Sequence , Amino Acid Substitution/genetics , Glutamic Acid/genetics , Humans , Microtubules/genetics , Microtubules/metabolism , Microtubules/pathology , Molecular Sequence Data , Phosphoprotein Phosphatases/metabolism , Phosphorylation , Polymerase Chain Reaction , Protein Conformation , Protein Phosphatase 2 , Serine/genetics , Structure-Activity Relationship , Threonine/genetics , tau Proteins/chemistry , tau Proteins/physiology
8.
Pneumologie ; 54(2): 49-53, 2000 Feb.
Article in German | MEDLINE | ID: mdl-10731756

ABSTRACT

The aim of the present investigation was to evaluate the diagnostic accuracy of positron emission tomography with 18-fluoro-2-deoxyglucose (FDG-PET) in the detection of recurrent lung cancer. PET was performed using an ECAT ART scanner (Siemens CTI) after i.v. injection of 220 +/- 50 MBq 18FDG. PET data were analysed by visual interpretation of coronal, sagittal and transversal slices. PET scans were interpreted independently by two experienced nuclear medicine physicians without prior knowledge of the results of other imaging studies or clinical data. 40 patients (= 41 cases) who had undergone primarily curative tumour treatment, were evaluated. In 29 of 35 cases with recurrent tumour, diagnosis was verified by pathologic means. FDG-PET correctly identified tumour recurrence in 34/35 cases. In 5/6 cases without prevent tumour recurrence PET gave true negative results. The overall accuracy of FDG-PET was 39/41 = 95% (95%-confidence interval 83-99%). FDG-PET shows high diagnostic accuracy in detecting recurrent lung cancer in patients with prior curative tumour treatment, but cannot substitute the need for pathological diagnosis.


Subject(s)
Carcinoma, Bronchogenic/diagnostic imaging , Carcinoma, Non-Small-Cell Lung/diagnostic imaging , Carcinoma, Small Cell/diagnostic imaging , Fluorodeoxyglucose F18 , Lung Neoplasms/diagnostic imaging , Neoplasm Recurrence, Local/diagnostic imaging , Tomography, Emission-Computed , Aged , Carcinoma, Bronchogenic/pathology , Carcinoma, Bronchogenic/therapy , Carcinoma, Non-Small-Cell Lung/pathology , Carcinoma, Non-Small-Cell Lung/therapy , Carcinoma, Small Cell/pathology , Carcinoma, Small Cell/therapy , Combined Modality Therapy , Female , Humans , Lung Neoplasms/pathology , Lung Neoplasms/therapy , Male , Middle Aged , Neoplasm Recurrence, Local/pathology , Neoplasm Recurrence, Local/therapy , Neoplasm Staging
9.
J Biol Chem ; 275(8): 5512-20, 2000 Feb 25.
Article in English | MEDLINE | ID: mdl-10681530

ABSTRACT

The human AC133 antigen and mouse prominin are structurally related plasma membrane proteins. However, their tissue distribution is distinct, with the AC133 antigen being found on hematopoietic stem and progenitor cells and prominin on various epithelial cells. To determine whether the human AC133 antigen and mouse prominin are orthologues or distinct members of a protein family, we examined the human epithelial cell line Caco-2 for the possible expression of the AC133 antigen. By both immunofluorescence and immunoprecipitation, the AC133 antigen was found to be expressed on the surface of Caco-2 cells. Interestingly, immunoreactivity for the AC133 antigen, but not its mRNA level, was down-regulated upon differentiation of Caco-2 cells. The AC133 antigen was specifically located at the apical rather than basolateral plasma membrane. An apical localization of the AC133 antigen was also observed in various human embryonic epithelia including the neural tube, gut, and kidney. Electron microscopy revealed that, within the apical plasma membrane of Caco-2 cells, the AC133 antigen was confined to microvilli and absent from the planar, intermicrovillar regions. This specific subcellular localization did not depend on an epithelial phenotype, because the AC133 antigen on hematopoietic stem cells, as well as that ectopically expressed in fibroblasts, was selectively found in plasma membrane protrusions. Hence, the human AC133 antigen shows the features characteristic of mouse prominin in epithelial and transfected non-epithelial cells, i.e. a selective association with apical microvilli and plasma membrane protrusions, respectively. Conversely, flow cytometry of murine CD34(+) bone marrow progenitors revealed the cell surface expression of prominin. Taken together, the data strongly suggest that the AC133 antigen is the human orthologue of prominin.


Subject(s)
Cell Membrane/metabolism , Epithelial Cells/metabolism , Glycoproteins/metabolism , Peptides/metabolism , AC133 Antigen , Animals , Antigens, CD , Blotting, Northern , CHO Cells , Caco-2 Cells , Cell Membrane/immunology , Cricetinae , Down-Regulation , Embryo, Mammalian/metabolism , Flow Cytometry , Glycoproteins/genetics , Glycoproteins/immunology , Hematopoietic Stem Cells/metabolism , Humans , Immunohistochemistry , Membrane Glycoproteins/chemistry , Mice , Microscopy, Electron , Peptides/genetics , Peptides/immunology , RNA, Messenger/metabolism , Time Factors , Tissue Distribution , Transfection
10.
Hum Mol Genet ; 9(1): 27-34, 2000 Jan 01.
Article in English | MEDLINE | ID: mdl-10587575

ABSTRACT

The disks of vertebrate photoreceptors are produced by outgrowths of the plasma membrane. Hence genes that encode retinal proteins targeted to plasma membrane protrusions represent candidates for inherited retinal degenerations. One such candidate is the gene encoding human prominin (mouse)-like 1 (PROML1, previously known as AC133 antigen) which belongs to the prominin family of 5-transmembrane domain proteins. Murine prominin (prom) shows a strong preference for plasma membrane protrusions in a variety of epithelial cells whereas PROML1 is expressed in retinoblastoma cell lines and adult retina. In the present study, molecular genetic analyses of a pedigree segregating for autosomal recessive retinal degeneration indicated that the affected individuals were homozygous for a nucleotide 1878 deletion in PROML1. This alteration is predicted to result in a frameshift at codon 614 with premature termination of translation. Expression of a similar prom deletion mutant in CHO cells indicated that the truncated protein does not reach the cell surface. Immunocytochemistry revealed that prom is concentrated in the plasma membrane evaginations at the base of the outer segments of rod photoreceptors. These findings suggest that loss of prominin causes retinal degeneration, possibly because of impaired generation of the evaginations and/or impaired conversion of the evaginations to disks.


Subject(s)
Frameshift Mutation , Glycoproteins/genetics , Glycoproteins/metabolism , Peptides/genetics , Peptides/metabolism , Retinal Degeneration/genetics , AC133 Antigen , Animals , Antigens, CD , Cell Membrane/metabolism , Chromosomes, Human, Pair 4 , Consanguinity , Female , Gene Expression Regulation , Genetic Markers , Glycoproteins/immunology , Humans , India , Male , Mice , Mice, Inbred Strains , Pedigree , Peptides/immunology , Polydactyly/genetics , Rod Cell Outer Segment/metabolism
11.
Med Klin (Munich) ; 94(4): 224-7, 1999 Apr 15.
Article in German | MEDLINE | ID: mdl-10373759

ABSTRACT

The value of FDG-PET in oncology is currently investigated in clinical studies. There is only limited information on the usefulness of FDG-PET in the evaluation of distant metastases of lung cancer. The purpose of the present prospective investigation was to determine the diagnostic accuracy of FDG-PET in the detection of brain metastases of lung cancer. After intravenous injection of 220 +/- 50 MBq F-18-deoxyglucose PET acquisition was carried out using an ECAT ART scanner (CTI Siemens). Images were reconstructed using a filtered backprojection with a Hanning filter. PET data were analyzed by visual interpretation of coronal, sagittal and transversal slices. PET scans were interpreted by two experienced nuclear medicine physicians without prior knowledge of the results of other imaging studies or clinical data. Between March 1997 and July 1998 whole-body PET was performed in 417 patients with suspected lung cancer. 402 patients were used for statistical analysis. Based on conventional brain imaging with CT (occasionally MRI), brain metastases were suspected in 17 patients (prevalence 4.2%). For FDG-PET alone, sensitivity was 82% (14/17) and specificity 38% (14/37). Therefore, diagnostic accuracy of FDG-PET in detection of brain metastases was 93.5%. The low specificity of FDG-PET can be explained by reduced tracer uptake mainly due to brain infarction or vascular encephalopathy in this group of elderly patients. Our results indicate that due to its low specificity FDG-PET is not useful for the evaluation of brain metastases in the primary staging of patients with lung cancer.


Subject(s)
Brain Neoplasms/diagnostic imaging , Brain Neoplasms/secondary , Fluorodeoxyglucose F18/therapeutic use , Lung Neoplasms/diagnostic imaging , Tomography, Emission-Computed/methods , Adult , Aged , Female , Humans , Male , Middle Aged , Sensitivity and Specificity
12.
J Cell Sci ; 112 ( Pt 7): 1023-33, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10198284

ABSTRACT

Prominin is a recently identified polytopic membrane protein expressed in various epithelial cells, where it is selectively associated with microvilli. When expressed in non-epithelial cells, prominin is enriched in plasma membrane protrusions. This raises the question of whether the selective association of prominin with microvilli in epithelial cells is solely due to its preference for, and stabilization in, plasma membrane protrusions, or is due to both sorting to the apical plasma membrane domain and subsequent enrichment in plasma membrane protrusions. To investigate this question, we have generated stably transfected MDCK cells expressing either full-length or C-terminally truncated forms of mouse prominin. Confocal immunofluorescence and domain-selective cell surface biotinylation experiments on transfected MDCK cells grown on permeable supports demonstrated the virtually exclusive apical localization of prominin at steady state. Pulse-chase experiments in combination with domain-selective cell surface biotinylation showed that newly synthesized prominin was directly targeted to the apical plasma membrane domain. Immunoelectron microscopy revealed that prominin was confined to microvilli rather than the planar region of the apical plasma membrane. Truncation of the cytoplasmic C-terminal tail of prominin impaired neither its apical cell surface expression nor its selective retention in microvilli. Both the apical-specific localization of prominin and its selective retention in microvilli were maintained when MDCK cells were cultured in low-calcium medium, i.e. in the absence of tight junctions. Taken together, our results show that: (i) prominin contains dual targeting information, for direct delivery to the apical plasma membrane domain and for the enrichment in the microvillar subdomain; and (ii) this dual targeting does not require the cytoplasmic C-terminal tail of prominin and still occurs in the absence of tight junctions. The latter observation suggests that entry into, and retention in, plasma membrane protrusions may play an important role in the establishment and maintenance of the apical-basal polarity of epithelial cells.


Subject(s)
Cell Membrane/metabolism , Epithelial Cells/metabolism , Membrane Glycoproteins/metabolism , Microvilli/metabolism , AC133 Antigen , Animals , Antigens, CD , Cell Line , Glycoproteins , Golgi Apparatus/metabolism , Immunoblotting , Immunohistochemistry , Kidney/metabolism , Kidney/ultrastructure , Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase/metabolism , Mice , Microscopy, Confocal , Microscopy, Immunoelectron , Models, Biological , Peptides , Recombinant Proteins/metabolism , Sequence Homology, Amino Acid , Tight Junctions/metabolism , Time Factors , Transfection
13.
EMBO J ; 17(12): 3277-89, 1998 Jun 15.
Article in English | MEDLINE | ID: mdl-9628865

ABSTRACT

Chromogranin B (CgB, secretogranin I) is a secretory granule matrix protein expressed in a wide variety of endocrine cells and neurons. Here we generated transgenic mice expressing CgB under the control of the human cytomegalovirus promoter. Northern and immunoblot analyses, in situ hybridization and immunocytochemistry revealed that the exocrine pancreas was the tissue with the highest level of ectopic CgB expression. Upon subcellular fractionation of the exocrine pancreas, the distribution of CgB in the various fractions was indistinguishable from that of amylase, an endogenous constituent of zymogen granules. Immunogold electron microscopy of pancreatic acinar cells showed co-localization of CgB with zymogens in Golgi cisternae, condensing vacuoles/immature granules and mature zymogen granules; the ratio of immunoreactivity of CgB to zymogens being highest in condensing vacuoles/immature granules. CgB isolated from zymogen granules of the pancreas of the transgenic mice aggregated in a mildly acidic (pH 5.5) milieu in vitro, suggesting that low pH-induced aggregation contributed to the observed concentration of CgB in condensing vacuoles. Our results show that a neuroendocrine-regulated secretory protein can be sorted to exocrine secretory granules in vivo, and imply that a key feature of CgB sorting in the trans-Golgi network of neuroendocrine cells, i.e. its aggregation-mediated concentration in the course of immature secretory granule formation, also occurs in exocrine cells although secretory protein sorting in these cells is thought to occur largely in the course of secretory granule maturation.


Subject(s)
Chromogranins/metabolism , Endocrine System/metabolism , Exocrine Glands/metabolism , Animals , Chromogranin B , Chromogranins/analysis , Chromogranins/genetics , Cytoplasmic Granules/metabolism , Endocrine System/cytology , Enzyme Precursors , Exocrine Glands/cytology , Golgi Apparatus/ultrastructure , Hydrogen-Ion Concentration , Immunohistochemistry , Mice , Mice, Inbred C57BL , Mice, Transgenic , Pancreas/chemistry , Pancreas/cytology , Pancreas/metabolism , Transgenes/genetics , Vacuoles/physiology
14.
EMBO J ; 17(4): 1107-19, 1998 Feb 16.
Article in English | MEDLINE | ID: mdl-9463388

ABSTRACT

Nup116p and Nup100p are highly related yeast GLFG nucleoporins, but only Nup116p is stoichiometrically bound to Gle2p, a previously identified mRNA export factor. A short Gle2p-binding sequence within Nup116p (GLEBS; residues 110-166) is sufficient and necessary to anchor Gle2p at the nuclear pores, whereas the carboxy-terminal domain of Nup116p mediates its own nuclear pore complex (NPC) association. The GLEBS is evolutionarily conserved and found in rat/Xenopus Nup98 and an uncharacterized Caenorhabditis elegans ORF, but is absent from Nup100p. When the GLEBS is deleted from Nup116p, Gle2p dissociates from the nuclear envelope and clusters of herniated nuclear pores form. When the GLEBS is inserted into Nup100p, Nup100p-GLEBS complements both the thermosensitive and NPC-herniated phenotype of nup116- cells, and Gle2p is retargeted concomitantly to the NPCs. Thus, the in vivo function of Gle2p is strictly coupled to the short GLEBS within Nup116p which links this putative mRNA transport factor to the nuclear pores.


Subject(s)
Conserved Sequence , Fungal Proteins/metabolism , Membrane Proteins/metabolism , Nuclear Matrix-Associated Proteins , Nuclear Pore Complex Proteins , Nuclear Proteins/metabolism , Nucleocytoplasmic Transport Proteins , RNA, Messenger/metabolism , Saccharomyces cerevisiae Proteins , Schizosaccharomyces pombe Proteins , Amino Acid Sequence , Biological Transport/genetics , Cell Nucleus/genetics , Cell Nucleus/metabolism , Fungal Proteins/genetics , Membrane Proteins/genetics , Molecular Sequence Data , Mutagenesis, Insertional , Nuclear Envelope/genetics , Nuclear Proteins/genetics , Protein Structure, Tertiary , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , Saccharomyces cerevisiae , Signal Transduction , Staphylococcal Protein A/genetics
15.
Behav Processes ; 43(2): 115-23, 1998 May.
Article in English | MEDLINE | ID: mdl-24896000

ABSTRACT

Dominance relationships were studied between marked or otherwise individually recognizable male green swordtails in a creek at Lake Catemaco and in a tributary of the Rio Atoyac (Veracruz, Mexico). The Atoyac population is unique because of a high degree of polymorphism, including both macromelanophore spotting and a micromelanophore tailspot pattern. During the dry season males living in the same area maintained a linear social hierarchy for periods of many days. The subordinate males settled down either in the same home ranges or in home ranges largely overlapping with that of dominant males. Although dominant males untiringly chased the subordinate males away, they returned persistently and achieved the status of non-tolerated satellites. Females were less stationary and presumably passed through many male home ranges during their feeding activities. The data clearly demonstrate that green swordtails live in complex social systems in which male-male competition and probably also female mate choice are likely to be essential factors for individual reproductive success.

16.
Proc Natl Acad Sci U S A ; 94(23): 12425-30, 1997 Nov 11.
Article in English | MEDLINE | ID: mdl-9356465

ABSTRACT

Using a new mAb raised against the mouse neuroepithelium, we have identified and cDNA-cloned prominin, an 858-amino acid-containing, 115-kDa glycoprotein. Prominin is a novel plasma membrane protein with an N-terminal extracellular domain, five transmembrane segments flanking two short cytoplasmic loops and two large glycosylated extracellular domains, and a cytoplasmic C-terminal domain. DNA sequences from Caenorhabditis elegans predict the existence of a protein with the same features, suggesting that prominin is conserved between vertebrates and invertebrates. Prominin is found not only in the neuroepithelium but also in various other epithelia of the mouse embryo. In the adult mouse, prominin has been detected in the brain ependymal layer, and in kidney tubules. In these epithelia, prominin is specific to the apical surface, where it is selectively associated with microvilli and microvilli-related structures. Remarkably, upon expression in CHO cells, prominin is preferentially localized to plasma membrane protrusions such as filopodia, lamellipodia, and microspikes. These observations imply that prominin contains information to be targeted to, and/or retained in, plasma membrane protrusions rather than the planar cell surface. Moreover, our results show that the mechanisms underlying targeting of membrane proteins to microvilli of epithelial cells and to plasma membrane protrusions of non-epithelial cells are highly related.


Subject(s)
Epithelial Cells/metabolism , Membrane Glycoproteins/metabolism , Membrane Proteins/metabolism , AC133 Antigen , Amino Acid Sequence , Animals , Antigens, CD , Biological Transport , CHO Cells , Cricetinae , Cytoplasmic Granules/metabolism , DNA, Complementary/genetics , DNA, Complementary/isolation & purification , Epithelial Cells/cytology , Glycoproteins , Membrane Glycoproteins/genetics , Membrane Proteins/genetics , Mice , Molecular Sequence Data , Peptides , Rats
17.
EMBO J ; 16(11): 3256-71, 1997 Jun 02.
Article in English | MEDLINE | ID: mdl-9214641

ABSTRACT

An essential cellular factor for nuclear mRNA export called Mex67p which has homologous proteins in human and Caenorhabditis elegans was identified through its genetic interaction with nucleoporin Nup85p. In the thermosensitive mex67-5 mutant, poly(A)+ RNA accumulates in intranuclear foci shortly after shift to the restrictive temperature, but NLS-mediated nuclear protein import is not inhibited. In vivo, Mex67p tagged with green fluorescent protein (GFP) is found at the nuclear pores, but mutant mex67-5-GFP accumulates in the cytoplasm. Upon purification of poly(A)+ RNA derived from of UV-irradiated yeast cells, Mex67p, but not nucleoporins Nup85p and Nup57p, was crosslinked to mRNA. In a two-hybrid screen, a putative RNA-binding protein with RNP consensus motifs was found to interact with the Mex67p carboxy-terminal domain. Thus, Mex67p is likely to participate directly in the export of mRNA from the nucleus to the cytoplasm.


Subject(s)
Cell Nucleus/metabolism , Nuclear Pore Complex Proteins , Nuclear Proteins/metabolism , Nucleocytoplasmic Transport Proteins , RNA, Messenger/metabolism , RNA-Binding Proteins/metabolism , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/genetics , Amino Acid Sequence , Biological Transport , Cell Compartmentation , Cell Nucleus/ultrastructure , Cloning, Molecular , Cytoplasm/metabolism , DNA Mutational Analysis , Molecular Sequence Data , Mutation , Nuclear Envelope/metabolism , Nuclear Envelope/ultrastructure , Nuclear Proteins/genetics , Polymerase Chain Reaction , Porins/genetics , Porins/metabolism , Protein Binding , RNA-Binding Proteins/genetics , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae/ultrastructure , Sequence Analysis, DNA , Sequence Homology, Amino Acid
18.
Mech Dev ; 69(1-2): 71-81, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9486532

ABSTRACT

Cell differentiation often involves changes in cell polarity. In this study we show that neuroepithelial cells, the progenitors of all neurons and macroglial cells of the vertebrate central nervous system, downregulate the polarized delivery to the apical and basolateral plasma membrane domains during development. Upon infection of the neuroepithelium of mouse embryos with fowl plague virus (FPV), polarized delivery of the viral envelope hemagglutinin, an apical marker, occurred at the neural plate stage (E8), but was downregulated at the open neural tube stage (E9). Upon infection with vesicular stomatitis virus, the viral envelope G protein, a basolateral marker, showed an unpolarized delivery not only at the open neural tube stage, but already at the neural plate stage. These results show that a progressive downregulation of plasma membrane polarity of neuroepithelial cells precedes neural tube closure and the onset of neurogenesis.


Subject(s)
Cell Polarity/physiology , Membrane Glycoproteins , Nervous System/embryology , Neural Tube Defects , Neurons/cytology , Animals , Cell Membrane/virology , Down-Regulation , Embryo, Mammalian/physiology , Embryo, Mammalian/virology , Embryonic Development , Epithelial Cells/metabolism , Epithelial Cells/ultrastructure , Epithelial Cells/virology , Female , Hemagglutinins, Viral/metabolism , Influenza A virus , Mice , Neurons/ultrastructure , Pregnancy , Vesicular stomatitis Indiana virus , Viral Envelope Proteins/metabolism
19.
Dev Biol ; 180(2): 664-79, 1996 Dec 15.
Article in English | MEDLINE | ID: mdl-8954735

ABSTRACT

Neuroepithelial cells can generate nonepithelial cells, the neurons. Here we have investigated, for chick and mouse embryos, the epithelial character of neuroepithelial cells in the context of neurogenesis by examining the presence of molecular components of tight junctions during the transition from the neural plate to the neural tube. Immunoreactivity for occludin, a transmembrane protein specific to tight junctions, was detected at the apical end of the lateral membrane of neuroepithelial cells throughout the chick neural plate. During neural tube closure, occludin disappeared from all neuroepithelial cells. Correspondingly, the addition of horseradish peroxidase to the apical side of the neuroepithelium by injection into the amniotic cavity of mouse embryos revealed the presence of functional tight junctions in the neural plate (Embryonic Day 8), but not the neural tube (Embryonic Day 9). In contrast to occludin, expression of ZO-1, a peripheral membrane protein of tight junctions, increased from the neural plate to the neural tube stage, also being confined to the apical end of the lateral neuroepithelial cell membrane. This localization coincided with that of N-cadherin, whose expression increased concomitantly with the disappearance of occludin. We propose that the loss of tight junctions from neuroepithelial cells reflects an overall decrease in their epithelial nature, which precedes the generation of neurons.


Subject(s)
Gene Expression Regulation, Developmental , Membrane Proteins/biosynthesis , Nervous System/embryology , Neurons/physiology , Phosphoproteins/biosynthesis , Tight Junctions/physiology , Amnion , Animals , Cadherins/biosynthesis , Cell Differentiation , Chick Embryo , Epithelial Cells , Epithelium/physiology , Epithelium/ultrastructure , Mice , Mice, Inbred Strains , Microscopy, Immunoelectron , Nervous System/cytology , Nervous System/ultrastructure , Neurons/cytology , Neurons/ultrastructure , Occludin , Species Specificity , Tubulin/biosynthesis , Zonula Occludens-1 Protein
20.
Ann Surg ; 224(2): 204-12, 1996 Aug.
Article in English | MEDLINE | ID: mdl-8757385

ABSTRACT

BACKGROUND/AIMS: After proctectomy for low rectal cancer and straight coloanal reconstruction, the main causes for increased daily stool frequency, urgency, and incontinence are the limited capacity and distensibility of the anastomosed colic segment in the pelvis. The authors postulated that a pedunculated (preserving the nerve) ileocecal interpositional graft (cecum-reservoir) placed between the sigmoid colon and the anal canal would greatly reduce these inconveniences. METHODS: The authors evaluated the safety, defecation quality, and anorectal physiology of such a neorectum in 20 consecutive patients with rectal carcinoma between 5 and 10 cm above the anal verge who underwent total mesorectal excision. RESULTS: No perioperative morbidity related to the technique and no mortality was observed in these 20 patients. Six months after the operation, 16 patients showed excellent and 4 patients good defecation quality, with maximal tolerable volumes, compliance, and mean colonic transit times comparable to age- and gender-matched healthy volunteers. In addition, anal resting pressure was decreased, squeeze pressure was maintained, and the rectoanal inhibitory reflex remained positive in 80%. CONCLUSIONS: The cecum-reservoir as a neorectum, using an intact neurovascular colonic segment, is a safe technique, providing excellent defecation quality. It enables a nearly normal physiologic anorectal function, which is already seen 6 months postoperatively.


Subject(s)
Cecum/surgery , Ileum/surgery , Rectal Neoplasms/surgery , Aged , Aged, 80 and over , Anastomosis, Surgical/adverse effects , Anastomosis, Surgical/methods , Defecation , Female , Humans , Male , Manometry , Middle Aged , Postoperative Complications/epidemiology , Rectum/physiology
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