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Other Publications (165)
- Experimental Cell Research
- Archives of Medical Research
- Genetics
- Development (Cambridge, England)
- Radiology
- Physiological Genomics
- Genome / National Research Council Canada = Génome / Conseil National De Recherches Canada
- Journal of Immunology (Baltimore, Md. : 1950)
- Prevention Science : the Official Journal of the Society for Prevention Research
- Methods in Enzymology
- Annals of Internal Medicine
- Journal of Clinical Microbiology
- Genetics
- AJR. American Journal of Roentgenology
- Journal of Long-term Effects of Medical Implants
- Journal of Long-term Effects of Medical Implants
- Journal of Long-term Effects of Medical Implants
- Journal of Long-term Effects of Medical Implants
- Journal of Long-term Effects of Medical Implants
- Journal of Back and Musculoskeletal Rehabilitation
- Preventive Medicine
- Journal of Neuroscience Research
- Journal of Immunology (Baltimore, Md. : 1950)
- The International Journal of Developmental Biology
- Current Treatment Options in Cardiovascular Medicine
- Journal of Plant Physiology
- Genetics
- Journal of Acquired Immune Deficiency Syndromes (1999)
- Journal of Wound, Ostomy, and Continence Nursing : Official Publication of The Wound, Ostomy and Continence Nurses Society / WOCN
- Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology
- Molecular Biology of the Cell
- Genome / National Research Council Canada = Génome / Conseil National De Recherches Canada
- Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology
- The American Journal of Pathology
- Gene Expression Patterns : GEP
- The Practising Midwife
- The International Journal of Developmental Biology
- The Practising Midwife
- Methods in Molecular Medicine
- Journal of the American Society of Nephrology : JASN
- The Journal of Urology
- The International Journal of Developmental Biology
- The Journal of Clinical Investigation
- Immunology
- Neuroscience Letters
- Genesis (New York, N.Y. : 2000)
- Clinical Lymphoma
- Cancer Biology & Therapy
- Cardiovascular Ultrasound
- Journal of Leukocyte Biology
- Journal of Virology
- Prehospital Emergency Care : Official Journal of the National Association of EMS Physicians and the National Association of State EMS Directors
- Journal of Immunology (Baltimore, Md. : 1950)
- The Journal of Reproductive Medicine
- Arthritis Research & Therapy
- Genomics
- Proceedings of the National Academy of Sciences of the United States of America
- Proceedings of the National Academy of Sciences of the United States of America
- Clinical Cancer Research : an Official Journal of the American Association for Cancer Research
- Journal of Virology
- Joint Commission Journal on Quality and Patient Safety / Joint Commission Resources
- Cell Biochemistry and Biophysics
- Emergency Radiology
- Molecular and Cellular Biology
- Journal of General Internal Medicine
- FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology
- Journal of Child Neurology
- Current Molecular Medicine
- Physiology (Bethesda, Md.)
- International Journal of Stroke : Official Journal of the International Stroke Society
- Alzheimer's & Dementia : the Journal of the Alzheimer's Association
- CSH Protocols
- Gene Expression Patterns : GEP
- Chemico-biological Interactions
- Experimental and Molecular Pathology
- Developmental Dynamics : an Official Publication of the American Association of Anatomists
- BJU International
- Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America
- European Journal of Cardiovascular Prevention and Rehabilitation : Official Journal of the European Society of Cardiology, Working Groups on Epidemiology & Prevention and Cardiac Rehabilitation and Exercise Physiology
- BJU International
- Journal of General Internal Medicine
- Journal of Shoulder and Elbow Surgery / American Shoulder and Elbow Surgeons ... [et Al.]
- Ecology
- Archives of Disease in Childhood. Fetal and Neonatal Edition
- Molecular and Cellular Biology
- The Journal of Urology
- The Journal of Pediatrics
- Development (Cambridge, England)
- The Journal of Antimicrobial Chemotherapy
- Nature
- Organogenesis
- Yeast (Chichester, England)
- Journal of the American Society of Echocardiography : Official Publication of the American Society of Echocardiography
- Clinical Cancer Research : an Official Journal of the American Association for Cancer Research
- Archives of Internal Medicine
- Mechanisms of Development
- Development (Cambridge, England)
- Developmental Biology
- Journal of the American Chemical Society
- Molecular Ecology
- Mechanisms of Development
- BJU International
- Acta Cytologica
- The Journal of Rheumatology
- Cytometry. Part A : the Journal of the International Society for Analytical Cytology
- Science (New York, N.Y.)
- Development (Cambridge, England)
- Journal of Nuclear Medicine Technology
- Advance for Nurse Practitioners
- Organogenesis
- Developmental Dynamics : an Official Publication of the American Association of Anatomists
- The International Journal of Developmental Biology
- Cancer Immunology, Immunotherapy : CII
- Environmental Management
- Annals of Clinical Biochemistry
- Journal of Nuclear Medicine Technology
- Methods in Molecular Biology (Clifton, N.J.)
- The Journal of Antimicrobial Chemotherapy
- Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology
- Chest
- Environment International
- Journal of Christian Nursing : a Quarterly Publication of Nurses Christian Fellowship
- Organic & Biomolecular Chemistry
- The International Journal of Developmental Biology
- Applied and Environmental Microbiology
- Microcirculation (New York, N.Y. : 1994)
- Journal of Nuclear Medicine Technology
- The Journal of Emergency Medicine
- Annals of Emergency Medicine
- Journal of Skin Cancer
- Organic Letters
- Journal of Wound, Ostomy, and Continence Nursing : Official Publication of The Wound, Ostomy and Continence Nurses Society / WOCN
- Respiratory Physiology & Neurobiology
- Journal of the American Society of Nephrology : JASN
- American Journal of Medical Genetics. Part A
- American Journal of Physiology. Heart and Circulatory Physiology
- Seminars in Cell & Developmental Biology
- Clinical Cancer Research : an Official Journal of the American Association for Cancer Research
- Journal of Clinical Sleep Medicine : JCSM : Official Publication of the American Academy of Sleep Medicine
- Beilstein Journal of Organic Chemistry
- Journal of Epidemiology and Community Health
- Pain
- Journal of Wound, Ostomy, and Continence Nursing : Official Publication of The Wound, Ostomy and Continence Nurses Society / WOCN
- Environmental Health Perspectives
- Academic Radiology
- Advanced Materials (Deerfield Beach, Fla.)
- JAMA : the Journal of the American Medical Association
- Archives of Disease in Childhood
- Developmental Dynamics : an Official Publication of the American Association of Anatomists
- Developmental Biology
- Biomedical Engineering Online
- Journal of Orthodontics
- Angiogenesis
- The Journal of Pharmacology and Experimental Therapeutics
- Prehospital Emergency Care : Official Journal of the National Association of EMS Physicians and the National Association of State EMS Directors
- American Family Physician
- The American Journal of Cardiology
- The International Journal of Oral & Maxillofacial Implants
- Radiology
- The Journal of School Health
- Academic Radiology
- Environmental Science & Technology
- Seminars in Fetal & Neonatal Medicine
- PloS One
- Nutrición Hospitalaria
Articles by Elizabeth Jones in JoVE
Other articles by Elizabeth Jones on PubMed
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Consumption of Fruits, Vegetables, Soft Drinks, and High-fat-containing Snacks Among Mexican Children on the Mexico-U.S. Border
Archives of Medical Research.
Jan-Feb, 2002 |
Pubmed ID: 11825635 The recommended diet for children would promote health, support growth, and prevent risk of disease. Diets high in fruits and vegetables demonstrate a strong and consistent pattern for decreasing the risk for many cancers and providing benefits against cardiovascular disease, diabetes, obesity, and stroke. The purpose of this study was to assess fruit, vegetable, soft drink, and high-fat-containing snack consumption among fifth- and ninth-grade children attending public schools in the northeastern Mexican state of Baja California.
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Annexin IV (Xanx-4) Has a Functional Role in the Formation of Pronephric Tubules
Development (Cambridge, England).
Apr, 2002 |
Pubmed ID: 11923205 Vertebrate kidney organogenesis is characterised by the successive formation of the pronephros, the mesonephros and the metanephros. The pronephros is the first to form and is the functional embryonic kidney of lower vertebrates; although it is vestigial in higher vertebrates, it is a necessary precursor for the other kidney types. The Xenopus pronephros is a simple paired organ; each nephron consists of a single large glomus, one set of tubules and a single duct. The simple organisation of the pronephros and the amenability of Xenopus laevis embryos to manipulation make the Xenopus pronephros an attractive system in which to study organogenesis. It has been shown that pronephric tubules can be induced to form in presumptive ectodermal tissue by treatment with RA and activin. We have used this system in a subtractive hybridisation screen that resulted in the cloning of Xenopus laevis annexin IV (Xanx-4). Xanx-4 transcripts are specifically located to the developing pronephric tubules, and the protein to the luminal surface of these tubules. Temporal expression shows zygotic transcription is upregulated at the time of pronephric tubule specification and persists throughout pronephric development. The temporal and spatial expression pattern of Xanx-4 suggests it may have a role in pronephric tubule development. Overexpression of Xanx-4 yields no apparent phenotype, but Xanx-4 depletion, using morpholinos, produces a shortened, enlarged tubule phenotype. The phenotype observed can be rescued by co-injection of Xanx-4 mRNA. Although the function of annexins is not yet clear, studies have suggested a role for annexins in a number of cellular processes. Annexin IV has been shown to have an inhibitory role in the regulation of epithelial calcium-activated chloride ion conductance. The enlarged pronephric tubule phenotype observed may be attributed to incorrect modulation of exocytosis, membrane plasticity or ion channels and/or water homeostasis. In this study, we demonstrate an in vivo role for annexin IV in the development of the pronephric tubules in Xenopus laevis.
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The Hypertrophic Heart Rat: a New Normotensive Model of Genetic Cardiac and Cardiomyocyte Hypertrophy
Physiological Genomics.
2002 |
Pubmed ID: 11948289 We describe a new line of rats with inherited cardiomyocyte and ventricular hypertrophy. From a second-generation cross of spontaneously hypertensive and Fischer 344 rats, we selected for low blood pressure and either high or low echocardiographic left ventricular (LV) mass over four generations to establish the hypertrophic heart rat (HHR) and normal heart rat (NHR) lines, respectively. After 13 generations of inbreeding, HHR had significantly greater (P < 0.0001) LV mass-to-body weight ratio (2.68 g/kg, SE 0.14) than NHR matched for age (1.94 g/kg, SE 0.02) or body weight (2.13 g/kg, SE 0.03). The isolated cardiomyocytes of HHR were significantly (P < 0.0001) longer and wider (161 microm, SE 0.83; 35.6 microm, SE 2.9) than NHR (132 microm, SE 1.2; 29.5 microm, SE 0.35). Telemetric 24-h recordings of mean arterial pressure revealed no significant differences between HHR and NHR. The HHR offers a new model of primary cardiomyocyte hypertrophy with normal blood pressure in which to examine genotypic causes and pathogenetic mechanisms of hypertrophy and its complications.
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An Enhanced Molecular Marker Based Genetic Map of Perennial Ryegrass (Lolium Perenne) Reveals Comparative Relationships with Other Poaceae Genomes
Genome / National Research Council Canada = Génome / Conseil National De Recherches Canada.
Apr, 2002 |
Pubmed ID: 11962626 A molecular-marker linkage map has been constructed for perennial ryegrass (Lolium perenne L.) using a one-way pseudo-testcross population based on the mating of a multiple heterozygous individual with a doubled haploid genotype. RFLP, AFLP, isoenzyme, and EST data from four collaborating laboratories within the International Lolium Genome Initiative were combined to produce an integrated genetic map containing 240 loci covering 811 cM on seven linkage groups. The map contained 124 codominant markers, of which 109 were heterologous anchor RFLP probes from wheat, barley, oat, and rice, allowing comparative relationships between perennial ryegrass and other Poaceae species to be inferred. The genetic maps of perennial ryegrass and the Triticeae cereals are highly conserved in terms of synteny and colinearity. This observation was supported by the general agreement of the syntenic relationships between perennial ryegrass, oat, and rice and those between the Triticeae and these species. A lower level of synteny and colinearity was observed between perennial ryegrass and oat compared with the Triticeae, despite the closer taxonomic affinity between these species. It is proposed that the linkage groups of perennial ryegrass be numbered in accordance with these syntenic relationships, to correspond to the homoeologous groups of the Triticeae cereals.
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Toll-like Receptor (TLR)2 and TLR4 in Human Peripheral Blood Granulocytes: a Critical Role for Monocytes in Leukocyte Lipopolysaccharide Responses
Journal of Immunology (Baltimore, Md. : 1950).
May, 2002 |
Pubmed ID: 11971020 Leukocyte responsiveness to LPS is dependent upon CD14 and receptors of the Toll-like receptor (TLR) family. Neutrophils respond to LPS, but conflicting data exist regarding LPS responses of eosinophils and basophils, and expression of TLRs at the protein level in these granulocyte lineages has not been fully described. We examined the expression of TLR2, TLR4, and CD14 and found that monocytes expressed relatively high levels of cell surface TLR2, TLR4, and CD14, while neutrophils also expressed all three molecules, but at low levels. In contrast, basophils expressed TLR2 and TLR4 but not CD14, while eosinophils expressed none of these proteins. Tested in a range of functional assays including L-selectin shedding, CD11b up-regulation, IL-8 mRNA generation, and cell survival, neutrophils responded to LPS, but eosinophils and basophils did not. In contrast to previous data, we found, using monocyte depletion by negative magnetic selection, that neutrophil responses to LPS were heavily dependent upon the presence of a very low level of monocytes, and neutrophil survival induced by LPS at 22 h was monocyte dependent. We conclude that LPS has little role in the regulation of peripheral blood eosinophil and basophil function, and that, even in neutrophils, monocytes orchestrate many previously observed leukocyte LPS response patterns.
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Role of the Unfolded Protein Response Pathway in Regulation of INO1 and in the Sec14 Bypass Mechanism in Saccharomyces Cerevisiae
Genetics.
Sep, 2002 |
Pubmed ID: 12242221 INO1, encoding inositol 1-phosphate synthase, is the most highly regulated of a class of genes containing the repeated element, UAS(INO), in their promoters. Transcription of UAS(INO)-containing genes is modulated by the availability of exogenous inositol and by signals generated by alteration of phospholipid metabolism. The unfolded protein response (UPR) pathway also is involved in INO1 expression and the ire1Delta and hac1Delta mutants are inositol auxotrophs. We examined the role of the UPR in transmitting a signal generated in response to inositol deprivation and to alteration of phospholipid biosynthesis created in the sec14(ts) cki1Delta genetic background. We report that the UPR is required for sustained high-level INO1 expression in wild-type strains, but not for transient derepression in response to inositol deprivation. Moreover, the UPR is not required for expression or regulation of INO1 in response to the change in lipid metabolism that occurs in the sec14(ts) cki1Delta genetic background. Thus, the UPR signal transduction pathway is not involved directly in transcriptional regulation of INO1 and other UAS(INO)-containing genes. However, we discovered that inactivation of Sec14p leads to activation of the UPR, and that sec14 cki1 strains exhibit defective vacuolar morphology, suggesting that the mechanism by which the cki1Delta mutation suppresses the growth and secretory defect of sec14 does not fully restore wild-type morphology. Finally, synthetic lethality involving sec14 and UPR mutations suggests that the UPR plays an essential role in survival of sec14 cki1 strains.
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Selective Roles for Toll-like Receptor (TLR)2 and TLR4 in the Regulation of Neutrophil Activation and Life Span
Journal of Immunology (Baltimore, Md. : 1950).
May, 2003 |
Pubmed ID: 12734376 Neutrophil responses to commercial LPS, a dual Toll-like receptor (TLR)2 and TLR4 activator, are regulated by TLR expression, but are amplified by contaminating monocytes in routine cell preparations. Therefore, we investigated the individual roles of TLR2 and TLR4 in highly purified, monocyte-depleted neutrophil preparations, using selective ligands (TLR2, Pam(3)CysSerLys(4) and Staphylococcus aureus peptidoglycan; TLR4, purified LPS). Activation of either TLR2 or TLR4 caused changes in adhesion molecule expression, respiratory burst (alone, and synergistically with fMLP), and IL-8 generation, which was, in part, dependent upon p38 mitogen-activated protein kinase signaling. Neutrophils also responded to Pam(3)CysSerLys(4) and purified LPS with down-regulation of the chemokine receptor CXCR2 and, to a lesser extent, down-regulation of CXCR1. TLR4 was the principal regulator of neutrophil survival, and TLR2 signals showed relatively less efficacy in preventing constitutive apoptosis over short time courses. TLR4-mediated neutrophil survival depended upon signaling via NF-kappa B and mitogen-activated protein kinase cascades. Prolonged neutrophil survival required both TLR4 activation and the presence of monocytes. TLR4 activation of monocytes was associated with the release of neutrophil survival factors, which was not evident with TLR2 activation, and TLR2 activation in monocyte/neutrophil cocultures did not prevent late neutrophil apoptosis. Thus, TLRs are important regulators of neutrophil activation and survival, with distinct and separate roles for TLR2 and TLR4 in neutrophil responses. TLR4 signaling presents itself as a pharmacological target that may allow therapeutic modulation of neutrophil survival by direct and indirect mechanisms at sites of inflammation.
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Isolation and Growth Factor Inducibility of the Xenopus Laevis Lmx1b Gene
The International Journal of Developmental Biology.
May, 2003 |
Pubmed ID: 12755330 This paper reports the cloning of the full length Xenopus laevis Lmx1b gene, Xlmx1b. Xlmx1b is a LIM homeodomain protein with high conservation to homologues identified in human, mouse, hamster and chick. In situ hybridisation and RT-PCR analysis showed that Xlmx1b has a specific temporal expression pattern which can be separated into three main spatial domains. An Xlmx1b probe hybridized to regions of the nervous system from stage 13 onwards; these regions included the placodes and otic vesicles, the eye and specific sets of neurons. Sectioning of in situ hybridised embryos confirmed the location of transcripts as discreet regions of staining in ventrolateral regions of the neural tube. From stage 27, transcripts could be detected in the capsule of pronephric glomus. Finally, transcripts were detected by Northern blot analysis in the developing fore and hind limbs. Xlmx1b transcripts were also detected by Northern blot analysis in eye, brain, muscle and mesonephros tissue in metamorphosing tadpoles. RT-PCR analysis showed that zygotic expression of Xlmx1b is initiated at stage 10.5 and the temporal sequence of Xlmx1b expression is identical in both neural and presumptive pronephros regions. The effects of the growth factors activin A, retinoic acid (RA) and basic fibroblast growth factor (bFGF) on the regulation of Xlmx1b were also studied. Xlmx1b was found to be upregulated by activin A and RA inhibited this upregulation in a concentration dependant manner. In contrast, bFGF had no effect on the regulation of Xlmx1b.
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Aortic Source of Brain Embolism
Current Treatment Options in Cardiovascular Medicine.
Jul, 2003 |
Pubmed ID: 12777199 Aortic arch atheroma has more recently been identified as an independent risk factor for ischemic stroke. Initially, this was a result of careful autopsy observations, then followed by a series of in vivo studies in which aortic arch atheroma was identified by transesophageal echocardiography. The association of aortic arch atheroma with ischemic stroke is most likely causal, given that the stroke risk increases with increasing thickness of arch atheroma. There is quite a sharp increase in stroke risk for atheroma of 4 mm or greater compared with lesser thicknesses. The clinical diagnosis is suggested when transient ischemic attack or ischemic stroke has occurred in which no obvious cardiac or arterial source of embolism is found. The presence of aortic arch atheroma is usually detected by transesophageal echocardiography and sometimes by magnetic resonance imaging or computed tomography. There is uncertainty about clinical management, particularly for secondary prevention. Options include the use of antiplatelet agents, anticoagulants, thrombolysis, or surgery. The latter two options have only been described rarely in case reports. Of the less invasive approaches, combination antiplatelet therapy with aspirin and clopidogrel is favored, or the use of warfarin. The Aortic arch Related Cerebral Hazard (ARCH) trial is being conducted to determine which of these is more effective in minimizing a composite outcome cluster of ischemic stroke, intracranial hemorrhage, myocardial infarction, peripheral embolism, or vascular death. Other more general management strategies should include reasonably aggressive risk factor control with blood pressure and lipid-lowering therapies and, if indicated, careful diabetic control.
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The Sec1/Munc18 Protein, Vps33p, Functions at the Endosome and the Vacuole of Saccharomyces Cerevisiae
Molecular Biology of the Cell.
Jun, 2004 |
Pubmed ID: 15047864 The Sec1/Munc18 (SM) family of proteins is thought to impart compartmental specificity to vesicle fusion reactions. Here we report characterization of Vps33p, an SM family member previously thought to act exclusively at the vacuolar membrane with the vacuolar syntaxin Vam3p. Vacuolar morphology of vps33Delta cells resembles that of cells lacking both Vam3p and the endosomal syntaxin Pep12p, suggesting that Vps33p may function with these syntaxins at the vacuole and the endosome. Consistent with this, vps33 mutants secrete the Golgi precursor form of the vacuolar hydrolase CPY into the medium. We also demonstrate that Vps33p acts at other steps, for vps33 mutants show severe defects in endocytosis at the late endosome. At the endosome, Vps33p and other class C members exist as a complex with Vps8p, a protein previously known to act in transport between the late Golgi and the endosome. Vps33p also interacts with Pep12p, a known interactor of the SM protein Vps45p. High copy PEP7/VAC1 suppresses vacuolar morphology defects of vps33 mutants. These findings demonstrate that Vps33p functions at multiple trafficking steps and is not limited to action at the vacuolar membrane. This is the first report demonstrating the involvement of a single syntaxin with two SM proteins at the same organelle.
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QTL Analysis of Quantitative Resistance to Phytophthora Infestans (late Blight) in Tomato and Comparisons with Potato
Genome / National Research Council Canada = Génome / Conseil National De Recherches Canada.
Jun, 2004 |
Pubmed ID: 15190365 Quantitative trait loci (QTLs) for resistance to Phytophthora infestans (late blight) were mapped in tomato. Reciprocal backcross populations derived from cultivated Lycopersicon esculentum x wild Lycopersicon hirsutum (BC-E, backcross to L. esculentum; BC-H, backcross to L. hirsutum) were phenotyped in three types of replicated disease assays (detached-leaflet, whole-plant, and field). Linkage maps were constructed for each BC population with RFLPs. Resistance QTLs were identified on all 12 tomato chromosomes using composite interval mapping. Six QTLs in BC-E (lb1a, lb2a, lb3, lb4, lb5b, and lb11b) and two QTLs in BC-H (lb5ab and lb6ab) were most consistently detected in replicated experiments or across assay methods. Lycopersicon hirsutum alleles conferred resistance at all QTLs except lb2a. Resistance QTLs coincided with QTLs for inoculum droplet dispersal on leaves, a trait in L. hirsutum that may contribute to resistance, and dispersal was mainly associated with leaf resistance. Some P. infestans resistance QTLs detected in tomato coincided with chromosomal locations of previously mapped R genes and QTLs for resistance to P. infestans in potato, suggesting functional conservation of resistance within the Solanaceae.
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The Role of Interleukin-1beta in Direct and Toll-like Receptor 4-mediated Neutrophil Activation and Survival
The American Journal of Pathology.
Nov, 2004 |
Pubmed ID: 15509550 The regulation of systemic and local neutrophil activation is crucial to the clearance of infections and the successful resolution of inflammation without progress to tissue damage or disseminated inflammatory reactions. Using purified lipopolysaccharide (pLPS) and highly purified neutrophils, we have previously shown that Toll-like receptor 4 signaling is a potent neutrophil activator, but a poor stimulator of survival. In the presence of peripheral blood mononuclear cells (PBMCs), however, pLPS becomes a potent neutrophil survival factor. Interleukin (IL)-1beta has been identified as an important neutrophil activator and prosurvival cytokine, and is produced in abundance by LPS-stimulated PBMCs. We now show that IL-1beta fails to activate highly purified neutrophils or enhance their survival, but in the presence of PBMCs, IL-1beta induces neutrophil survival. We hypothesized that LPS-primed neutrophils might become responsive to IL-1beta, but were unable to demonstrate this. Moreover, IL-1ra failed to prevent pLPS + PBMC-dependent neutrophil survival. In studies of IL-1R1(-/-) mice, we found that LPS was still able to mediate neutrophil survival, and neutrophil survival was enhanced by the addition of monocytic cells. Thus an important paradigm of neutrophil regulation needs to be viewed in the context of a cellular network in which actions of IL-1beta on neutrophils are indirect and mediated by other cells.
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X-epilectin: a Novel Epidermal Fucolectin Regulated by BMP Signalling
The International Journal of Developmental Biology.
Dec, 2004 |
Pubmed ID: 15602698 This paper reports the cloning and characterisation of a new posterior epidermal marker, X-epilectin, in Xenopus laevis. This gene encodes for a fucolectin, which belongs to the lectin superfamily of carbohydrate binding proteins and specifically binds fucose residues. RT-PCR and in situ hybridisation show that the expression of this gene is switched on during gastrulation and up-regulated during neurula stages and found expressed ubiquitously throughout the epidermis. From tailbud stages, the expression is limited to the dorsal posterior region of the embryo, suggesting that X-epilectin expression is regulated along anteroposterior and dorsoventral gradients during development. In the adult, X-epilectin is mainly expressed in intestinal components, kidney, spinal cord and skin. The effects of growth factors on the regulation of X-epilectin were studied. Change of the fate of animal caps into cement gland or dorsal mesoderm induces a down-regulation of X-epilectin expression in explants treated respectively with ammonium chloride and activin A. We also show that X-epilectin expression is down-regulated by Noggin and tBR and that this effect is inhibited by BMP4 over-expression, suggesting X-epilectin expression is mediated by the BMP signalling pathway.
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Developmental Expression of Pod 1 in Xenopus Laevis
The International Journal of Developmental Biology.
2005 |
Pubmed ID: 15744669 The basic helix-loop-helix transcription factor, Pod 1, has been shown to be expressed in the mesenchyme of many developing mouse organs, including the heart, lungs and gut. In the kidneys of developing mice, Pod 1 is highly expressed in the condensing metanephric mesenchyme, differentiating and late stromal cells and in developing podocytes. We have obtained an EST (CF270487) which contains the Xenopus laevis Pod 1 sequence. Conceptual translation of the Xenopus laevis Pod 1 sequence shows approximately 85% similarity to other vertebrate homologues. RT-PCR indicates that expression is initiated at stage 13 and increases differentially in the developing pronephros compared to the whole embryo. RT-PCR of a kidney dissection at stage 42 shows higher expression in the glomus than in the tubule or duct. In situ hybridisation analysis at tail bud stages shows the anterior-most branchial arch and pronephric glomus are intensely stained. At stage 40, staining persists in the glomus and in the epicardium region of the heart. Adult organ analysis shows expression is highest in the rectum and the spleen, with significant expression in the duodenum, heart, kidney, lungs, pancreas, skin, liver and muscle.
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Age-dependent Incidence, Time Course, and Consequences of Thymic Renewal in Adults
The Journal of Clinical Investigation.
Apr, 2005 |
Pubmed ID: 15776111 Homeostatic regulation of T cells involves an ongoing balance of new T cell generation, peripheral expansion, and turnover. The recovery of T cells when this balance is disrupted provides insight into the mechanisms that govern homeostasis. In a long-term, single cohort study, we assessed the role of thymic function after autologous transplant in adults, correlating serial computed tomography imaging of thymic size with concurrent measurements of peripheral CD4(+) T cell populations. We established the age-dependent incidence, time course, and duration of thymic enlargement in adults and demonstrated that these changes were correlated with peripheral recovery of naive CD45RA(+)CD62L(+) and signal-joint TCR rearrangement excision circle-bearing CD4(+) populations with broad TCR diversity. Furthermore, we demonstrated that renewed thymopoiesis was critical for the restoration of peripheral CD4(+) T cell populations. This recovery encompassed the recovery of normal CD4(+) T cell numbers, a low ratio of effector to central memory cells, and a broad repertoire of TCR Vbeta diversity among these memory cells. These data define the timeline and consequences of renewal of adult thymopoietic activity at levels able to quantitatively restore peripheral T cell populations. They further suggest that structural thymic regrowth serves as a basis for the regeneration of peripheral T cell populations.
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Regulation of Human Neutrophil Chemokine Receptor Expression and Function by Activation of Toll-like Receptors 2 and 4
Immunology.
May, 2005 |
Pubmed ID: 15819701 Neutrophil chemokine receptor expression can be altered by exposure to Toll-like receptor (TLR) agonists, a process that is thought to have the potential to localize neutrophils to sites of infection. In order to investigate this process in more detail, we examined the regulation of highly pure neutrophil CXCR1 and CXCR2 expression and function by selective agonists of TLR2 (Pam(3)CSK(4)) and TLR4 (lipopolysaccharide, LPS). CXCR1 and CXCR2 were down-regulated by TLR engagement. CXCR2 loss was more rapid and showed a dependence upon soluble helper molecules (LPS binding protein and CD14) that was not evident for CXCR1, suggesting differential coupling of LPS signalling to CXCR1 and CXCR2 loss. However, TLR engagement in highly pure neutrophils did not result in complete loss of chemokine receptors, and LPS-treated neutrophils remained able to mount a respiratory burst to CXCL8 and CXCL1, and were able to migrate towards CXCL8 in assays of under-agarose chemotaxis. Thus, although treatment of purified human neutrophils with TLR2 and TLR4 agonists modifies chemokine receptor expression, remaining receptors remain functionally competent.
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The Human DEK Proto-oncogene is a Senescence Inhibitor and an Upregulated Target of High-risk Human Papillomavirus E7
Journal of Virology.
Nov, 2005 |
Pubmed ID: 16254365 The human DEK proto-oncogene is a nucleic acid binding protein with suspected roles in human carcinogenesis, autoimmune disease, and viral infection. Intracellular DEK functions, however, are poorly understood. In papillomavirus-positive cervical cancer cells, downregulation of viral E6/E7 oncogene expression results in cellular senescence. We report here the specific repression of DEK message and protein levels in senescing human papillomavirus type 16- (HPV16-) and HPV18-positive cancer cell lines as well as in primary cells undergoing replicative senescence. Cervical cancer cell senescence was partially overcome by DEK overexpression, and DEK overexpression was sufficient for extending the life span of primary keratinocytes, supporting critical roles for this molecule as a senescence regulator. In order to determine whether DEK is a bona fide HPV oncogene target in primary cells, DEK expression was monitored in human keratinocytes transduced with HPV E6 and/or E7. The results identify high-risk HPV E7 as a positive DEK regulator, an activity that is not shared by low-risk HPV E7 protein. Experiments in mouse embryo fibroblasts recapitulated the observed E7-mediated DEK induction and demonstrated that both basal and E7-induced regulation of DEK expression are controlled by the retinoblastoma protein family. Taken together, our results suggest that DEK upregulation may be a common event in human carcinogenesis and may reflect its senescence inhibitory function.
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Distal Enhancer Elements Transcribe Intergenic RNA in the IL-10 Family Gene Cluster
Journal of Immunology (Baltimore, Md. : 1950).
Dec, 2005 |
Pubmed ID: 16301651 The IL-10 gene and homologs IL-19, IL-20, and IL-24 are expressed within a highly conserved 145-kb cytokine gene cluster. Like the Th2 IL-4 cytokine gene cluster, it is feasible that there is coordinate regulation of these cytokines by distal regulatory elements spanning the locus. We initiated a search to characterize regulatory elements within the IL-10 family locus and present data herein on a conserved 40-kb region between the IL-19 and IL-10 genes. We map the location of 17 DNase I-hypersensitive sites in different murine T cell populations and identify three enhancer elements, which function in T cells in vitro. Two of these enhancer elements, located 9 kb upstream and 6.45 kb downstream of IL-10, display cell-specific function in the Th1-Th2 cell clones AE7 and D10 and also exhibit basic promoter activity. The downstream element, IL-10CNS+6.45, binds AP-1 in the absence of NFAT and expresses intergenic RNA in a Th2-specific manner, further validating its role as a Th2-specific enhancer/promoter element. We show that the five most highly conserved noncoding sequences in the 40-kb region transcribe intergenic RNA; four of these regions possess promoter activity in vitro that could account for the expression of these transcripts. Hence, we speculate that these novel regulatory elements in the IL-10 family gene locus function via an intermediate regulatory RNA.
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Analysis of Fcgamma Receptor Haplotypes in Rheumatoid Arthritis: FCGR3A Remains a Major Susceptibility Gene at This Locus, with an Additional Contribution from FCGR3B
Arthritis Research & Therapy.
2006 |
Pubmed ID: 16356189 The Fcgamma receptors play important roles in the initiation and regulation of many immunological and inflammatory processes, and genetic variants (FCGR) have been associated with numerous autoimmune and infectious diseases. The data in rheumatoid arthritis (RA) are conflicting and we previously demonstrated an association between FCGR3A and RA. In view of the close molecular proximity with FCGR2A, FCGR2B and FCGR3B, additional polymorphisms within these genes and FCGR haplotypes were examined to refine the extent of association with RA. Biallelic polymorphisms in FCGR2A, FCGR2B and FCGR3B were examined for association with RA in two well characterized UK Caucasian and North Indian/Pakistani cohorts, in which FCGR3A genotyping had previously been undertaken. Haplotype frequencies and linkage disequilibrium were estimated across the FCGR locus and a model-free analysis was performed to determine association with RA. This was followed by regression analysis, allowing for phase uncertainty, to identify the particular haplotype(s) that influences disease risk. Our results reveal that FCGR2A, FCGR2B and FCGR3B were not associated with RA. The haplotype with the strongest association with RA susceptibility was the FCGR3A-FCGR3B 158V-NA2 haplotype (odds ratio 3.18, 95% confidence interval 1.13-8.92 [P = 0.03] for homozygotes compared with all genotypes). The association was stronger in the presence of nodules (odds ratio 5.03, 95% confidence interval 1.44-17.56; P = 0.01). This haplotype was also more common in North Indian/Pakistani RA patients than in control individuals, but not significantly so. Logistic regression analyses suggested that FCGR3A remained the most significant gene at this locus. The increased association with an FCGR3A-FCGR3B haplotype suggests that other polymorphic variants within FCGR3A or FCGR3B, or in linkage disequilibrium with this haplotype, may additionally contribute to disease pathogenesis.
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Comparative Genomic and Expression Analysis of the Conserved NTPDase Gene Family in Xenopus
Genomics.
Mar, 2006 |
Pubmed ID: 16380227 The purines, ATP and adenosine, are important signaling molecules in the nervous system. ATP is sequentially degraded to adenosine by the ectonucleotidase proteins. The NTPDase (or CD39) family is a subfamily of these enzymes, which consists of nine members in mammals. In Xenopus embryos, we have shown that ATP, and its antagonist adenosine, regulate the rundown of swimming and we therefore proposed that ectonucleotidase proteins are key regulators of locomotor activity. Here, we report the cloning of all nine members of the NTPDase family in Xenopus laevis and Xenopus tropicalis. Our phylogenetic analysis shows that this family is highly conserved between the frog species and also during vertebrate evolution. In the adult frog, NTPDase genes are broadly expressed. During development, all NTPDase genes, except for NTPDase8, are expressed and display a distinct specific expression pattern, suggesting potentially different functions of these proteins during embryogenesis of X. laevis.
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Pbn1p: an Essential Endoplasmic Reticulum Membrane Protein Required for Protein Processing in the Endoplasmic Reticulum of Budding Yeast
Proceedings of the National Academy of Sciences of the United States of America.
Jan, 2006 |
Pubmed ID: 16418276 PBN1 was identified as a gene required for production of protease B (PrB) activity in Saccharomyces cerevisiae. PBN1 encodes an endoplasmic reticulum (ER)-localized, type I membrane glycoprotein and is essential for cell viability. To study the essential function(s) of Pbn1p, we constructed a strain with PBN1 under control of the GAL promoter. Depletion of Pbn1p in this strain abrogates processing of the ER precursor forms of PrB, Gas1p, and Pho8p. Depletion of Pbn1p does not affect exit of proprotease A or procarboxypeptidase Y from the ER, indicating that Pbn1p is not required for global exit from the ER. Depleting Pbn1p leads to a significant increase in the unfolded protein response pathway, accompanied by an expansion of bulk ER membrane, indicating that there is a defect in protein folding in the ER. pbn1-1, a nonlethal allele of PBN1, displays synthetic lethality with the ero1-1 allele (ERO1 is required for oxidation in the ER) and synthetic growth defects with the cne1Delta allele (CNE1 encodes calnexin). ER-associated degradation of a lumenal substrate, CPY*, is blocked in the absence of Pbn1p. These results suggest that Pbn1p is required for proper folding and/or the stability of a subset of proteins in the ER. Thus, Pbn1p is an essential chaperone-like protein in the ER of yeast.
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Dynamic Regulation of Pro- and Anti-inflammatory Cytokines by MAPK Phosphatase 1 (MKP-1) in Innate Immune Responses
Proceedings of the National Academy of Sciences of the United States of America.
Feb, 2006 |
Pubmed ID: 16461893 Engagement of Toll-like receptors (TLRs) on macrophages leads to activation of the mitogen-activated protein kinases (MAPKs), which contribute to innate immune responses. MAPK activity is regulated negatively by MAPK phosphatases (MKPs). MKP-1, the founding member of this family of dual-specificity phosphatases, has been implicated in regulating lipopolysaccharide (LPS) responses, but its role in TLR-mediated immune responses in vivo has not been defined. Here, we show that mice deficient in MKP-1 were highly susceptible to endotoxic shock in vivo, associated with enhanced production of proinflammatory cytokines TNF-alpha and IL-6 and an anti-inflammatory cytokine, IL-10. We further examined the regulation and function of MKP-1 in macrophages, a major cell type involved in endotoxic shock. MKP-1 was transiently induced by TLR stimulation through pathways mediated by both myeloid differentiation factor 88 (MyD88) and TIR domain-containing adaptor inducing IFN-beta (TRIF). MKP-1 deficiency led to sustained activation of p38 MAPK and c-Jun N-terminal kinase (JNK) in LPS-treated macrophages. In response to TLR signals, MKP-1-deficient macrophages produced 5- to 10-fold higher IL-10, which could be blocked by a p38 MAPK inhibitor. Thus, p38 MAPK plays a critical role in mediating IL-10 synthesis in TLR signaling. TNF-alpha was found to be more abundant in MKP-1-deficient macrophages within 2 hours of TLR stimulation, but its production was rapidly down-regulated by IL-10. Our studies demonstrate that MKP-1 attenuates the activities of p38 MAPK and JNK to regulate both pro- and anti-inflammatory cytokines in TLR signaling. These results highlight the complex mechanisms by which the MAPKs regulate innate immunity.
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Living in Three Dimensions: 3D Nanostructured Environments for Cell Culture and Regenerative Medicine
Cell Biochemistry and Biophysics.
2006 |
Pubmed ID: 16757822 Research focused on deciphering the biochemical mechanisms that regulate cell proliferation and function has largely depended on the use of tissue culture methods in which cells are grown on two-dimensional (2D) plastic or glass surfaces. However, the flat surface of the tissue culture plate represents a poor topological approximation of the more complex three-dimensional (3D) architecture of the extracellular matrix (ECM) and the basement membrane (BM), a structurally compact form of the ECM. Recent work has provided strong evidence that the highly porous nanotopography that results from the 3D associations of ECM and BM nanofibrils is essential for the reproduction of physiological patterns of cell adherence, cytoskeletal organization, migration, signal transduction, morphogenesis, and differentiation in cell culture. In vitro approximations of these nanostructured surfaces are therefore desirable for more physiologically mimetic model systems to study both normal and abnormal functions of cells, tissues, and organs. In addition, the development of 3D culture environments is imperative to achieve more accurate cell-based assays of drug sensitivity, high-throughput drug discovery assays, and in vivo and ex vivo growth of tissues for applications in regenerative medicine.
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Apoptosis Inhibition by the Human DEK Oncoprotein Involves Interference with P53 Functions
Molecular and Cellular Biology.
Oct, 2006 |
Pubmed ID: 16894028 The DEK proto-oncogene has been associated with human carcinogenesis-either as a fusion with the CAN nucleoporin protein or when transcriptionally upregulated. Mechanisms of intracellular DEK functions, however, have remained relatively unexplored. We have recently demonstrated that DEK expression is induced by the high-risk human papillomavirus (HPV) E7 protein in a manner which is dependent upon retinoblastoma protein function and have implicated DEK in the inhibition of cellular senescence. Additionally, overexpression of DEK resulted in significant life span extension of primary human keratinocytes. In order to determine whether DEK expression is required for cellular proliferation and/or survival, we monitored cellular responses to the knockdown of DEK in cancer and primary cells. The results indicate that DEK expression protects both HPV-positive cancer and primary human cells from apoptotic cell death. Cell death in response to DEK depletion was accompanied by increased protein stability and transcriptional activity of the p53 tumor suppressor and consequent upregulation of known p53 target genes such as p21CIP and Bax. Consistent with a possible role for p53 in DEK-mediated cell death inhibition, the p53-negative human osteosarcoma cell line SAOS-2 was resistant to the knockdown of DEK. Finally, expression of a dominant negative p53 miniprotein inhibited DEK RNA interference-induced p53 transcriptional induction, as well as cell death, thus directly implicating p53 activation in the observed apoptotic phenotype. These findings suggest a novel role for DEK in cellular survival, involving the destabilization of p53 in a manner which is likely to contribute to human carcinogenesis.
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Pediatric Epstein-Barr Virus-Associated Encephalitis: 10-Year Review
Journal of Child Neurology.
May, 2006 |
Pubmed ID: 16948923 Many neurologic manifestations of Epstein-Barr virus (EBV) infection have been documented, including encephalitis, aseptic meningitis, transverse myelitis, and Guillain-Barr?yndrome. These manifestations can occur alone or coincidentally with the clinical picture of infectious mononucleosis. Since 1994, The Hospital for Sick Children has maintained a prospective registry of all children admitted with acute encephalitis. This report summarizes all cases of Epstein-Barr virus-associated encephalitis compiled from 1994 to 2003. Twenty-one (6%) of 216 children, median age 13 years (range 3-17 years), in the Encephalitis Registry were identified as having evidence of Epstein-Barr virus infection. This evidence consisted of convincing Epstein-Barr virus serology and/or positive cerebrospinal fluid polymerase chain reaction (PCR). One patient had symptoms of classic infectious mononucleosis; all others had a nonspecific prodrome, including fever (n = 17; 81%) and headache (n = 14; 66%). Slightly less than half (n = 10; 48%) had seizures and often had electroencephalograms showing a slow background (n = 12; 57%). Many demonstrated cerebrospinal fluid pleocytosis (n = 17; 81%), and 71% (n = 15) had abnormal magnetic resonance imaging findings. Two patients died, 2 suffered mild deficits, and 16 were neurologically normal at follow-up. Most patients with Epstein-Barr virus encephalitis do not show typical symptoms of infectious mononucleosis. Establishing a diagnosis of Epstein-Barr virus encephalitis can be difficult, and, consequently, a combination of serologic and molecular techniques should be used when investigating a child with acute encephalitis. Most children make full recoveries, but residual neurologic sequelae and even death can and do occur. (J Child Neurol 2006;21:384-391; DOI 10.2310/7010.2006.00114).
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What Determines Blood Vessel Structure? Genetic Prespecification Vs. Hemodynamics
Physiology (Bethesda, Md.).
Dec, 2006 |
Pubmed ID: 17119151 Vascular network remodeling, angiogenesis, and arteriogenesis play an important role in the pathophysiology of ischemic cardiovascular diseases and cancer. Based on recent studies of vascular network development in the embryo, several novel aspects to angiogenesis have been identified as crucial to generate a functional vascular network. These aspects include specification of arterial and venous identity in vessels and network patterning. In early embryogenesis, vessel identity and positioning are genetically hardwired and involve neural guidance genes expressed in the vascular system. We demonstrated that, during later stages of embryogenesis, blood flow plays a crucial role in regulating vessel identity and network remodeling. The flow-evoked remodeling process is dynamic and involves a high degree of vessel plasticity. The open question in the field is how genetically predetermined processes in vessel identity and patterning balance with the contribution of blood flow in shaping a functional vascular architecture. Although blood flow is essential, it remains unclear to what extent flow is able to act on the developing cardiovascular system. There is significant evidence that mechanical forces created by flowing blood are biologically active within the embryo and that the level of mechanical forces and the type of flow patterns present in the embryo are able to affect gene expression. Here, we highlight the pivotal role for blood flow and physical forces in shaping the cardiovascular system.
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Aortic Arch Atheroma
International Journal of Stroke : Official Journal of the International Stroke Society.
May, 2006 |
Pubmed ID: 18706048 Severe atheroma of the aortic arch has now been established as an important risk factor and mechanism for stroke and peripheral embolism. The odds ratio for stroke or peripheral embolism in patients with severe arch atheroma is greater than four, and for mobile atheroma it is greater than 12. The prevalence of severe arch atheroma among patients presenting with acute ischaemic stroke, at over 20%, is in the same order as that of atrial fibrillation and carotid atherosclerosis. In patients with ischaemic stroke for which no cause has been identified, it is reasonable to determine as to whether they have severe arch atheroma by performing a transoesophageal echocardiogram. Recurrent stroke is common in patients with aortic arch atheroma that are thicker than 4 mm or with mobile components, particularly in the elderly, cigarette smokers, and those with hypertension or diabetes. Patients found to have severe atheroma are at high risk of recurrent events (14.2% per year) and may, therefore, need an aggressive secondary prevention strategy. Currently, there is uncertainty as to what this should be, but either combination antiplatelet therapy (aspirin plus clopidogrel) or anticoagulation with warfarin (target INR 2.0-3.0) are commonly used. Which of these is most effective will be evident after the completion of the aortic arch related cerebral hazard trial.
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GDNF Expression During Xenopus Development
Gene Expression Patterns : GEP.
Jan, 2007 |
Pubmed ID: 17049928 Glial cell line-derived neurotrophic factor (GDNF) has multiple roles in kidney morphogenesis, spermatogenesis, and neurogenesis during development. In this study, we report the cloning and expression pattern of Xenopus laevis GDNF. The X. laevis GDNF cDNA sequence has a complete open reading frame of 684 bases, predicting 227 amino acid residues at the protein level. Comparison of the X. laevis GDNF amino acid sequence with those of chick, human, mouse, rat and zebrafish indicates that X. laevis GDNF has 60%-52% and 75%-62% identity over the whole amino acid sequence and for the putative mature forms, respectively. All known functional motifs of GDNF were conserved in the X. laevis sequence. Temporal expression analysis by RT-PCR indicated that GDNF transcripts were first detectable at stage 12 at a low level, and gradually increased up to stage 22. From stage 24, the expression sharply increased and continued at a similar level as development progressed. Spatial expression analysis by whole-mount in situ hybridization showed that the GDNF mRNA was predominantly detected in somites, pronephros, pharyngeal arches, epibranchial placodes, digestive tract and some of the lateral line structure. These results suggest that this X. laevis gene is the orthologue for GDNF.
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Ultrasonographic Examination of the Synovial Fold of the Radiohumeral Joint
Journal of Shoulder and Elbow Surgery / American Shoulder and Elbow Surgeons ... [et Al.].
Sep-Oct, 2007 |
Pubmed ID: 17507252 This report describes the anatomy of the synovial fold of the radiohumeral joint and assesses its visibility by ultrasonography. Forty-nine fresh cadaver radiohumeral joints were examined by ultrasonography before and after intraarticular saline injection and then dissected. Digital photos were taken before and after the joint capsule was excised. The relative coverage of the radial head by the fold was calculated. Synovial folds were observed in all specimens. Forty-three had anterior and posterior lobes. The synovial fold covered an average of 28% of the radiocapitellar joint surface of the radial head. The sensitivity of the ultrasonography was 81%, 46%, and 85% from the anterior, lateral, and posterior aspects of the radiohumeral joint, respectively. Intraarticular saline injection improved the sensitivity to 96%, 67%, and 94%, respectively. The synovial fold is a consistent anatomic structure, and ultrasonography can be a useful preoperative diagnostic tool.
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Genetic Identity Determines Risk of Post-settlement Mortality of a Marine Fish
Ecology.
May, 2007 |
Pubmed ID: 17536412 Longitudinal sampling of four cohorts of Neopomacentrus filamentosus, a common tropical damselfish from Dampier Archipelago, Western Australia, revealed the evolution of size structure after settlement. Light traps collected premetamorphic individuals from the water column ("settlers") to establish a baseline for each cohort. Subsequently, divers collected benthic juveniles ("recruits") at 1-3-month intervals to determine the relative impacts of post-settlement mortality during the first three months. Growth trajectories for individual fish were back-calculated from otolith records and compared with nonlinear mixed-effects models. Size-selective mortality was detected in all cohorts with the loss of smaller, slower growing individuals. Three months after settlement, recruits showed significantly faster growth as juveniles, faster growth as larvae, and larger sizes as hatchlings. The timing and intensity of post-settlement selection differed among cohorts and was correlated with density at settlement. The cohort with the greatest initial abundance experienced the strongest selective mortality, with most of this mortality occurring between one and two months after settlement when juveniles began foraging at higher positions in the water column. Significant genetic structure was found between settlers and three-month-old recruits in this cohort as a result of natural selection that changed the frequency of mtDNA haplotypes measured at the control region. The extent of this genetic difference was enlarged or reduced by artificially manipulating the intensity of size-based selection, thus establishing a link between phenotype and haplotype. Sequence variation in the control region of the mitochondrial genome has been linked to mitochondrial efficiency and weight gain in other studies, which provides a plausible explanation for the patterns observed here.
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Regulation of SRC-3 Intercompartmental Dynamics by Estrogen Receptor and Phosphorylation
Molecular and Cellular Biology.
Oct, 2007 |
Pubmed ID: 17646391 The steroid receptor coactivator 3 gene (SRC-3) (AIB1/ACTR/pCIP/RAC3/TRAM1) is a p160 family transcription coactivator and a known oncogene. Despite its importance, the functional regulation of SRC-3 remains poorly understood within a cellular context. Using a novel combination of live-cell, high-throughput, and fluorescent microscopy, we report SRC-3 to be a nucleocytoplasmic shuttling protein whose intracellular mobility, solubility, and cellular localization are regulated by phosphorylation and estrogen receptor alpha (ERalpha) interactions. We show that both chemical inhibition and small interfering RNA reduction of the mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2 (MEK1/2) pathway induce a cytoplasmic shift in SRC-3 localization, whereas stimulation by epidermal growth factor signaling enhances its nuclear localization by inducing phosphorylation at T24, S857, and S860, known participants in the phosphocode that regulates SRC-3 activity. Accordingly, the cytoplasmic localization of a nonphosphorylatable SRC-3 mutant further supported these results. In the presence of ERalpha, U0126 also dramatically reduces (i) ligand-dependent colocalization of SRC-3 and ERalpha, (ii) the formation of ER-SRC-3 complexes in cell lysates, and (iii) SRC-3 targeting to a visible, ERalpha-occupied and -regulated prolactin promoter array. Taken together, these results indicate that phosphorylation coordinates SRC-3 coactivator function by linking the probabilistic formation of transient nuclear receptor-coactivator complexes with its molecular dynamics and cellular compartmentalization. Technically and conceptually, these findings have a new and broad impact upon evaluating mechanisms of action of gene regulators at a cellular system level.
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Vascular Remodeling of the Mouse Yolk Sac Requires Hemodynamic Force
Development (Cambridge, England).
Sep, 2007 |
Pubmed ID: 17720695 The embryonic heart and vessels are dynamic and form and remodel while functional. Much has been learned about the genetic mechanisms underlying the development of the cardiovascular system, but we are just beginning to understand how changes in heart and vessel structure are influenced by hemodynamic forces such as shear stress. Recent work has shown that vessel remodeling in the mouse yolk sac is secondarily effected when cardiac function is reduced or absent. These findings indicate that proper circulation is required for vessel remodeling, but have not defined whether the role of circulation is to provide mechanical cues, to deliver oxygen or to circulate signaling molecules. Here, we used time-lapse confocal microscopy to determine the role of fluid-derived forces in vessel remodeling in the developing murine yolk sac. Novel methods were used to characterize flows in normal embryos and in embryos with impaired contractility (Mlc2a(-/-)). We found abnormal plasma and erythroblast circulation in these embryos, which led us to hypothesize that the entry of erythroblasts into circulation is a key event in triggering vessel remodeling. We tested this by sequestering erythroblasts in the blood islands, thereby lowering the hematocrit and reducing shear stress, and found that vessel remodeling and the expression of eNOS (Nos3) depends on erythroblast flow. Further, we rescued remodeling defects and eNOS expression in low-hematocrit embryos by restoring the viscosity of the blood. These data show that hemodynamic force is necessary and sufficient to induce vessel remodeling in the mammalian yolk sac.
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Purine-mediated Signalling Triggers Eye Development
Nature.
Oct, 2007 |
Pubmed ID: 17960245 A conserved network of eye field transcription factors (EFTFs) underlies the development of the eye in vertebrates and invertebrates. To direct eye development, Pax6, a key gene in this network, interacts with genes encoding other EFTFs such as Rx1 and Six3 (refs 4-6). However, the mechanisms that control expression of the EFTFs remain unclear. Here we show that purine-mediated signalling triggers both EFTF expression and eye development in Xenopus laevis. Overexpression of ectonucleoside triphosphate diphosphohydrolase 2 (E-NTPDase2), an ectoenzyme that converts ATP to ADP, caused ectopic eye-like structures, with occasional complete duplication of the eye, and increased expression of Pax6, Rx1 and Six3. In contrast, downregulation of endogenous E-NTPDase2 decreased Rx1 and Pax6 expression. E-NTPDase2 therefore acts upstream of these EFTFs. To test whether ADP (the product of E-NTPDase2) might act to trigger eye development through P2Y1 receptors, selective in Xenopus for ADP, we simultaneously knocked down expression of the genes encoding E-NTPDase2 and the P2Y1 receptor. This could prevent the expression of Rx1 and Pax6 and eye formation completely. We next measured ATP release in the presumptive eye field, demonstrating a transient release of ATP at a time that could plausibly trigger (once converted to ADP) expression of the EFTFs. This surprising role for transient purine-mediated signalling in eye development may be widely conserved, because alterations to the locus of E-NTPDase2 on human chromosome 9 cause severe head and eye defects, including microphthalmia. Our results suggest a new mechanism for the initiation of eye development.
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A Functional Screen for Genes Involved in Xenopus Pronephros Development
Mechanisms of Development.
Jul, 2008 |
Pubmed ID: 18472403 In Xenopus, the pronephros is the functional larval kidney and consists of two identifiable components; the glomus, the pronephric tubules, which can be divided into four separate segments, based on marker gene expression. The simplicity of this organ, coupled with the fact that it displays the same basic organization and function as more complex mesonephros and metanephros, makes this an attractive model to study vertebrate kidney formation. In this study, we have performed a functional screen specifically to identify genes involved in pronephros development in Xenopus. Gain-of-function screens are performed by injecting mRNA pools made from a non-redundant X. tropicalis full-length plasmid cDNA library into X. laevis eggs, followed by sib-selection to identify the single clone that caused abnormal phenotypes in the pronephros. Out of 768 egg and gastrula stage cDNA clones, 31 genes, approximately 4% of the screened clones, affected pronephric marker expression examined by whole mount in situ hybridization or antibody staining. Most of the positive clones had clear expression patterns in pronephros and predicted/established functions highly likely to be involved in developmental processes. In order to carry out a more detailed study, we selected Sox7, Cpeb3, P53csv, Mecr and Dnajc15, which had highly specific expression patterns in the pronephric region. The over-expression of these five selected clones indicated that they caused pronephric abnormalities with different temporal and spatial effects. These results suggest that our strategy to identify novel genes involved in pronephros development was highly successful, and that this strategy is effective for the identification of novel genes involved in late developmental events.
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Comparative Phylogeography of Two Seastars and Their Ectosymbionts Within the Coral Triangle
Molecular Ecology.
Dec, 2008 |
Pubmed ID: 19067797 Repeated exposure and flooding of the Sunda and Sahul shelves during Pleistocene sea-level fluctuations is thought to have contributed to the isolation and diversification of sea-basin populations within the Coral Triangle. This hypothesis has been tested in numerous phylogeographical studies, recovering an assortment of genetic patterns that the authors have generally attributed to differences in larval dispersal capability or adult habitat specificity. This study compares phylogeographical patterns from mitochondrial COI sequences among two co-distributed seastars that differ in their adult habitat and dispersal ability, and two seastar ectosymbionts that differ in their degree of host specificity. Of these, only the seastar Linckia laevigata displayed a classical pattern of Indian-Pacific divergence, but with only moderate genetic structure (PhiCT = 0.067). In contrast, the seastarProtoreaster nodosus exhibited strong structure (PhiCT = 0.23) between Teluk Cenderawasih and the remainder of Indonesia, a pattern of regional structure that was echoed in L. laevigata (PhiCT = 0.03) as well as its obligate gastropod parasite Thyca crystallina (PhiCT = 0.04). The generalist commensal shrimp, Periclimenes soror showed little genetic structuring across the Coral Triangle. Despite species-specific phylogeographical patterns, all four species showed departures from neutrality that are consistent with massive range expansions onto the continental shelves as the sea levels rose, and that date within the Pleistocene epoch.Our results suggest that habitat differences may affect the manner in which species responded to Pleistocene sea-level fluctuations, shaping contemporary patterns of genetic structure and diversity.
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Breast-specific Gamma-imaging: Molecular Imaging of the Breast Using 99mTc-sestamibi and a Small-field-of-view Gamma-camera
Journal of Nuclear Medicine Technology.
Dec, 2009 |
Pubmed ID: 19914975 Breast-specific gamma-imaging (BSGI), also known as molecular breast imaging, is breast scintigraphy using a small-field-of-view gamma-camera and (99m)Tc-sestamibi. There are many different types of breast cancer, and many have characteristics making them challenging to detect by mammography and ultrasound. BSGI is a cost-effective, highly sensitive and specific technique that complements other imaging modalities currently being used to identify malignant lesions in the breast. Using the current Society of Nuclear Medicine guidelines for breast scintigraphy, Legacy Good Samaritan Hospital began conducting BSGI, breast scintigraphy with a breast-optimized gamma-camera. In our experience, optimal imaging has been conducted in the Breast Center by a nuclear medicine technologist. In addition, the breast radiologists read the BSGI images in correlation with the mammograms, ultrasounds, and other imaging studies performed. By modifying the current Society of Nuclear Medicine protocol to adapt it to the practice of breast scintigraphy with these new systems and by providing image interpretation in conjunction with the other breast imaging studies, our center has found BSGI to be a valuable adjunctive procedure in the diagnosis of breast cancer. The development of a small-field-of-view gamma-camera, designed to optimize breast imaging, has resulted in improved detection capabilities, particularly for lesions less than 1 cm. Our experience with this procedure has proven to aid in the clinical work-up of many of our breast patients. After reading this article, the reader should understand the history of breast scintigraphy, the pharmaceutical used, patient preparation and positioning, imaging protocol guidelines, clinical indications, and the role of breast scintigraphy in breast cancer diagnosis.
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Phase II Trial of Bevacizumab, Thalidomide, Docetaxel, and Prednisone in Patients with Metastatic Castration-resistant Prostate Cancer
Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology.
Apr, 2010 |
Pubmed ID: 20308663 PURPOSE We previously demonstrated that thalidomide appears to add to the activity of docetaxel in metastatic castration-resistant prostate cancer (CRPC). Phase II studies combining docetaxel with bevacizumab have had substantial antitumor activity. We hypothesized that the combination of docetaxel plus these antiangiogenic drugs with different targets would have substantial clinical activity. To explore safety and efficacy, this was tested in mice and in human patients. PATIENTS AND METHODS Preclinical efficacy of the combination therapy was evaluated in PC3 xenograft mice. Sixty patients with progressive metastatic CRPC received intravenous docetaxel and bevacizumab plus oral thalidomide and prednisone. The primary end point was a prostate-specific antigen (PSA) decline of > or = 50%. Secondary end points included time to progression, overall survival, and safety. Results In the mouse model, combination therapy of docetaxel, bevacizumab, and thalidomide inhibited tumor growth most effectively. In the clinical trial, 90% of patients receiving the combination therapy had PSA declines of > or = 50%, and 88% achieved a PSA decline of > or = 30% within the first 3 months of treatment. The median time to progression was 18.3 months, and the median overall survival was 28.2 months in this group with a Halabi-predicted survival of 14 months. While toxicities were manageable, all patients developed grade 3/4 neutropenia. CONCLUSION The addition of bevacizumab and thalidomide to docetaxel is a highly active combination with manageable toxicities. The estimated median survival is encouraging, given the generally poor prognosis of this patient population. These results suggest that definitive clinical trials combining antiangiogenic agent combinations with docetaxel are warranted to improve treatment outcomes for patients with metastatic CRPC.
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The Lysophosphatidic Acid (LPA) and Sphingosine-1-phosphate (S1P) Receptor Gene Families: Cloning and Comparative Expression Analysis in Xenopus Laevis
The International Journal of Developmental Biology.
2010 |
Pubmed ID: 20712001 Sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) are endogenous bioactive lipids which mediate a variety of biological cell responses such as cell proliferation, migration, differentiation and apoptosis. Their actions are mediated by binding to the G-protein-coupled endothelial differentiation gene (Edg) receptor subfamily, referred to as S1P1-5 and LPA1-5, and regulate a variety of signalling pathways involved in numerous physiological processes and pathological conditions. Their importance during embryogenesis has been demonstrated by the generation of knock-out mice and specific roles have been assigned to these receptors. However, potential functional redundancy and the lethality of some mutants have complicated functional analysis in these models. Here we report the cloning of the S1P and LPA receptors in Xenopus laevis and tropicalis. Phylogenetic analyses demonstrate the high level of conservation of these receptors between amphibian and other vertebrate species. We have conducted a comparative expression analysis of these receptors during development and in the adult frog, by both RT-PCR and whole mount in situ hybridisation. In particular, we show that S1P1, 2 and 5 display distinct embryonic specific expression patterns, suggesting potentially different developmental roles for these receptors, and therefore for their ligands, during amphibian embryogenesis.
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Stimulation of Methane Generation from Nonproductive Coal by Addition of Nutrients or a Microbial Consortium
Applied and Environmental Microbiology.
Nov, 2010 |
Pubmed ID: 20817801 Biogenic formation of methane from coal is of great interest as an underexploited source of clean energy. The goal of some coal bed producers is to extend coal bed methane productivity and to utilize hydrocarbon wastes such as coal slurry to generate new methane. However, the process and factors controlling the process, and thus ways to stimulate it, are poorly understood. Subbituminous coal from a nonproductive well in south Texas was stimulated to produce methane in microcosms when the native population was supplemented with nutrients (biostimulation) or when nutrients and a consortium of bacteria and methanogens enriched from wetland sediment were added (bioaugmentation). The native population enriched by nutrient addition included Pseudomonas spp., Veillonellaceae, and Methanosarcina barkeri. The bioaugmented microcosm generated methane more rapidly and to a higher concentration than the biostimulated microcosm. Dissolved organics, including long-chain fatty acids, single-ring aromatics, and long-chain alkanes accumulated in the first 39 days of the bioaugmented microcosm and were then degraded, accompanied by generation of methane. The bioaugmented microcosm was dominated by Geobacter sp., and most of the methane generation was associated with growth of Methanosaeta concilii. The ability of the bioaugmentation culture to produce methane from coal intermediates was confirmed in incubations of culture with representative organic compounds. This study indicates that methane production could be stimulated at the nonproductive field site and that low microbial biomass may be limiting in situ methane generation. In addition, the microcosm study suggests that the pathway for generating methane from coal involves complex microbial partnerships.
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Rheology of Embryonic Avian Blood
American Journal of Physiology. Heart and Circulatory Physiology.
Dec, 2011 |
Pubmed ID: 21963831 Shear stress, a mechanical force created by blood flow, is known to affect the developing cardiovascular system. Shear stress is a function of both shear rate and viscosity. While established techniques for measuring shear rate in embryos have been developed, the viscosity of embryonic blood has never been known but always assumed to be like adult blood. Blood is a non-Newtonian fluid, where the relationship between shear rate and shear stress is nonlinear. In this work, we analyzed the non-Newtonian behavior of embryonic chicken blood using a microviscometer and present the apparent viscosity at different hematocrits, different shear rates, and at different stages during development from 4 days (Hamburger-Hamilton stage 22) to 8 days (about Hamburger-Hamilton stage 34) of incubation. We chose the chicken embryo since it has become a common animal model for studying hemodynamics in the developing cardiovascular system. We found that the hematocrit increases with the stage of development. The viscosity of embryonic avian blood in all developmental stages studied was shear rate dependent and behaved in a non-Newtonian manner similar to that of adult blood. The range of shear rates and hematocrits at which non-Newtonian behavior was observed is, however, outside the physiological range for the larger vessels of the embryo. Under low shear stress conditions, the spherical nucleated blood cells that make up embryonic blood formed into small aggregates of cells. We found that the apparent blood viscosity decreases at a given hematocrit during embryonic development, not due to changes in protein composition of the plasma but possibly due to the changes in cellular composition of embryonic blood. This decrease in apparent viscosity was only visible at high hematocrit. At physiological values of hematocrit, embryonic blood viscosity did not change significantly with the stage of development.
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The Prevalence and Management of Low Back Pain Across Adulthood: Results from a Population-based Cross-sectional Study (the MUSICIAN Study)
Pain.
Jan, 2012 |
Pubmed ID: 21978663 The aim of the current study was to determine: the prevalence of low back pain (LBP) and associated disability; the frequency of consultation to general practice; whether there were differences in management by age. We conducted a cross-sectional population study in Aberdeen city and Cheshire County, UK. Participants were 15,272 persons aged 25 years and older. The 1-month period prevalence of LBP was 28.5%. It peaked at age 41-50 years, but at ages over 80 years was reported by 1 in 4 persons. Older persons were more likely to consult, and the prevalence of severe LBP continued to increase with age. Management by general practitioners differed by age of the patient. Older persons (> 70 vs ≤ 40 years) were more likely to only have been prescribed painkillers (odds ratio [OR] 1.74, 95% confidence interval [CI] 1.28-2.35) or only pain killers with other medications (OR 1.45, 95% CI 1.07-1.98). They were less likely to be prescribed physiotherapy or exercise (OR 0.63, 95% CI 0.46-0.85) or to be referred to a specialist (OR 0.77, 95% CI 0.57-1.04). Older persons were more likely to have previously received exercise therapy for pain, were less likely to be enthusiastic about receiving it now (P
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The Glycocalyx is Present As Soon As Blood Flow is Initiated and is Required for Normal Vascular Development
Developmental Biology.
Sep, 2012 |
Pubmed ID: 22820069 The glycocalyx, and the thicker endothelial surface layer (ESL), are necessary both for endothelial barrier function and for sensing mechanical forces in the adult. The goal of this study is to use a combination of imaging techniques to establish when the glycocalyx and endothelial surface layer form during embryonic development and to determine the biological significance of the glycocalyx layer during vascular development in quail embryos. Using transmission electron microscopy, we show that the glycocalyx layer is present as soon as blood flow starts (14 somites). The early endothelial glycocalyx (14 somites) lacks the distinct hair-like morphology that is present later in development (17 and 25 somites). The average thickness does not change significantly (14 somites, 182 nm ± 33 nm; 17 somites, 218 ± 30 nm; 25 somites, 212 ± 32 nm). The trapping of circulating fluorescent albumin was used to evaluate the development of the ESL. Trapped fluorescent albumin was first observed at 25 somites. In order to assess a functional role for the glycocalyx during development, we selectively degraded luminal glycosaminoglycans. Degradation of hyaluronan compromised endothelial barrier function and prevented vascular remodeling. Degradation of heparan sulfate down regulated the expression of shear-sensitive genes but does not inhibit vascular remodeling. Our findings show that the glycocalyx layer is present as soon as blood flow starts (14 somites). Selective degradations of major glycocalyx components were shown to inhibit normal vascular development, examined through morphology, vascular barrier function, and gene expression.
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LCL124, a Cationic Analog of Ceramide, Selectively Induces Pancreatic Cancer Cell Death by Accumulating in Mitochondria
The Journal of Pharmacology and Experimental Therapeutics.
Jan, 2013 |
Pubmed ID: 23086228 Treatment of pancreatic cancer that cannot be surgically resected currently relies on minimally beneficial cytotoxic chemotherapy with gemcitabine. As the fourth leading cause of cancer-related death in the United States with dismal survival statistics, pancreatic cancer demands new and more effective treatment approaches. Resistance to gemcitabine is nearly universal and appears to involve defects in the intrinsic/mitochondrial apoptotic pathway. The bioactive sphingolipid ceramide is a critical mediator of apoptosis initiated by a number of therapeutic modalities. It is noteworthy that insufficient ceramide accumulation has been linked to gemcitabine resistance in multiple cancer types, including pancreatic cancer. Taking advantage of the fact that cancer cells frequently have more negatively charged mitochondria, we investigated a means to circumvent resistance to gemcitabine by targeting delivery of a cationic ceramide (l-t-C6-CCPS [LCL124: ((2S,3S,4E)-2-N-[6'-(1″-pyridinium)-hexanoyl-sphingosine bromide)]) to cancer cell mitochondria. LCL124 was effective in initiating apoptosis by causing mitochondrial depolarization in pancreatic cancer cells but demonstrated significantly less activity against nonmalignant pancreatic ductal epithelial cells. Furthermore, we demonstrate that the mitochondrial membrane potentials of the cancer cells were more negative than nonmalignant cells and that dissipation of this potential abrogated cell killing by LCL124, establishing that the effectiveness of this compound is potential-dependent. LCL124 selectively accumulated in and inhibited the growth of xenografts in vivo, confirming the tumor selectivity and therapeutic potential of cationic ceramides in pancreatic cancer. It is noteworthy that gemcitabine-resistant pancreatic cancer cells became more sensitive to subsequent treatment with LCL124, suggesting that this compound may be a uniquely suited to overcome gemcitabine resistance in pancreatic cancer.
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Health Maintenance in Women
American Family Physician.
Jan, 2013 |
Pubmed ID: 23317023 The health maintenance examination is an opportunity to focus on disease prevention and health promotion. The patient history should include screening for tobacco use, alcohol misuse, intimate partner violence, and depression. Premenopausal women should receive preconception counseling and contraception as needed, and all women planning or capable of pregnancy should take 400 to 800 mcg of folic acid per day. High-risk sexually active women should be counseled on reducing the risk of sexually transmitted infections, and screened for chlamydia, gonorrhea, and syphilis. All women should be screened for human immunodeficiency virus. Adults should be screened for obesity and elevated blood pressure. Women 20 years and older should be screened for dyslipidemia if they are at increased risk of coronary heart disease. Those with sustained blood pressure greater than 135/80 mm Hg should be screened for type 2 diabetes mellitus. Women 55 to 79 years of age should take 75 mg of aspirin per day when the benefits of stroke reduction outweigh the increased risk of gastrointestinal hemorrhage. Women should begin cervical cancer screening by Papanicolaou test at 21 years of age, and if results have been normal, screening may be discontinued at 65 years of age or after total hysterectomy. Breast cancer screening with mammography may be considered in women 40 to 49 years of age based on patients' values, and potential benefits and harms. Mammography is recommended biennially in women 50 to 74 years of age. Women should be screened for colorectal cancer from 50 to 75 years of age. Osteoporosis screening is recommended in women 65 years and older, and in younger women with a similar risk of fracture. Adults should be immunized at recommended intervals according to guidelines from the Centers for Disease Control and Prevention.
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Prevalence of Obesity and Abdominal Obesity from Four to 16 Years Old Children Living in the Mexico-USA Border
Nutrición Hospitalaria.
Mar-Apr, 2013 |
Pubmed ID: 23822701 The prevalence of obesity among Mexicans is alarming in both the child and adult populations. The objective of this study was to determine the levels of overweight, obesity and abdominal obesity in pre-school (PS), elementary (ES), and middle high (MHS) public school children from Tijuana. From February to April of 2011, a bietapic random sample was selected by cluster method of 30 PS, 30 ES, and 30 MHS children. And a sample of 30 groups for each level was chosen. Twenty elementary teachers and eight graduate students were trained at one central location on how to take anthropometric measurements using a portable scale, a stadiometer, and a measuring tape to determine weight, height, and waist circumference. Body Mass Index values were computed and compared to age/gender BMI percentiles according to WHO criteria. Waist circumference for-age at the 90th percentile from NHANES III (Mexican-American) was used to define abdominal obesity. The sample was composed of 646 PS children, 961 ES children, and 1,095 MHS children. Their ages ranged from 4- 16 years. Results showed an overall prevalence of overweight and obesity in younger than 5y preschool children (> 2 SD) of 23.1%, in ≥ 5 y PS (> 1 SD) of 33.8%, in ES children of 46.3%, and in MHS children of 41.9%. Abdominal obesity in PS children was 18%, in ES children was 16.7%, and in MHS children was 15.2%. These results warrant immediate and comprehensive actions to prevent a critical public health problem in Mexico.
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