Exosomes in Breast Cancer Essays

Exosomes in Breast Cancer Essays Exosomes in Breast Cancer Essay Exosomes in Breast Cancer Essay History of exosomes find The bringing of lading between intracellular compartments within cells by agencies of cysts has been surveies for many old ages. However, the acknowledgment that extracellular release of assorted types of cysts has complex biological results in multiple systems, including pathological conditions such as malignant neoplastic disease, has merely become apparent in recent old ages. Amongst several types of secreted cysts, exosomes were first described in 1981 as microvesicles incorporating 5-nucleotidase activity that were secreted from neoplastic cell lines ( Trams, et al. , 1981 ) . A few old ages subsequently, the secernment of cysts of endocytic beginning by civilized reticulocytes was reported. At the clip, some writers believed late endosomes were transitional comdepartments and their content was non thought to be recycled back to the plasma membrane, but destined for debasement in lysosomes. Consequently, these consequences were controversial, and were considered to be basically membranes shed by deceasing cells in civilization. In 1989, nevertheless, surveies observed the merger between late endosomes and the plasma membrane in cytotoxic T lymphocytes ( Peters et al. , 1989 ) , which proved that late endosomes were non merely transitional sections. The maps of exosomes were non widely studied until 1996, when surveies by Raposo and co-workers described exosomes bearing functional MHC category II molecules secreted by B lymph cells, triping renewed involvement in the survey of these cysts ( Raposo, et al. , 1996 ) . At present, exosomes have been demonstrated to show in a figure of organic structure fluids, such as piss ( Pisitkun et al. , 2004 ) , breast milk ( Admyre et al. , 2007 ) , blood plasma ( Caby et al. , 2005 ) , bronchoalveolar lavage fluid ( Admyre et al. , 2003 ) , amnionic fluid ( Keller et al, 2007 ) and malignant gushs ( Andre et al. , 2002 ) , bespeaking their importance in vivo.This study will concentrate on exosomes, which should non be confused with other secreted membrane cysts such as microvesicles, thatmicrovesicles, that have distinguishable structural and biochemical belongingss. Biology and composing of Exosomes Exosomes are characterised as membrane cysts with a size scope of 50-100nm. They are from endosomal beginning and are secreted by most cell types, such as hematopoietic, lymphocytes, dendritic cells, epithelial ( Van Niel et al. , 2001 ) , reticulocytes ( Johnstone et al. , 1987 ) and many tumor cells.Exosomes are limited by a lipid bilayer and can be formed through inward budding of endosomal membranes, giving rise to intracellular multivesicular organic structures ( MVB ) that subsequently fuse with the plasma membrane, let go ofing the exosomes to the extracellular environment. They can besides be formed by direct outward budding from the plasma membrane.The transportation of constituents between different compartments in a cell involves bearer cysts that bud from one compartment and so merger with another compartment after going in the cytol. These classical lading cysts display their cytoplasmatic side of the compartment they form from on their outer surface and stay intracellul ar. In contrast membrane cysts that are secreted into the extracellular environment display the extracellular side of the membrane they form from at their outer surface, so in consequence they could be considered as a illumination version of a cell.The molecular composing of exosomes depends on the types of cells they derived from. 1 is a conventional representation of a canonical exosomes. The proteins listed are found in at least 30 % of different exosomes, and proteins besides marked with an star are present in at least 50 % of exosomes.The cytosolic Rab protein that promote exosomes docking and membrane merger are present on exosomes. Annexin I, II, V and VI, may play a function in modulating membrane cytoskeleton kineticss and membrane merger events. Some adhesion molecules such as intercellular adhesion molecule-1, CD146, CD9, MFG-E8, CD11a, are besides present on exosomes. Proteins that are involved in programmed cell death are present on exosomes such as Alix and galectin 3. Heat daze proteins Hsp70 and Hsp90 are besides present, they are up-regulated under emphasis state of affairss and aid with protein folding and protect cells from emphasis. Exosomes besides contain tetraspanins including CD9, CD63, CD81 and CD82. Some proteins are cell-type specific ; MHC category I molecules are present in exosomes from most cell types, while CD86 and MHC category II molecules are merely present in exosomes that are derived from antigen-presenting cells ( APCs ) . Similar to proteins, lipid composing on exosomes besides varies. The typical lipid composing of exosomes derived from mast cells includes lysophosphatidylcholine, sphingomyelin, phosphatidylcholine, phosphatidylserine and cholesterin, etc ( Schorey et al. , 2007 ) . Isolation of exosomes Exosomes can be isolated based on their size, denseness and biochemical belongingss. The most widely used method of sublimating exosomes from cell civilization supernatant uses a series of centrifugations to take dead cells and cell dusts ( normally 1000 x g, so 10,000 x g ) , followed by a concluding high velocity ultracentrifugation ( 100,000 x g ) to pellet the exosomes ( Thery et al. , 2002 ) . A filtration procedure ( 0.2 millimeter ) can besides take cells and dust. This procedure does non know apart between exosomes and other little cysts though, as a effect, exosomes must be identified utilizing other extra standards. Furthermore, exosomes have a specific denseness ( ranges from 1.13 gml-1 to 1.19 gml-1 ) and can be purified by floatation into a sucrose denseness gradient or by sucrosedeuterium oxide shock absorbers ( Thery et al. , 2006 ) .Another purification method is based on the size and utilizes chromatography ( Taylor et al. , 2006 ) . In add-on, exosomes can be isolat ed based on their membrane belongingss. Beads coated with an antibody against a protein known to be expressed on the exosome membrane can be added to the supernatant after cell depletion, for illustration anti-MHC category II magnetic Dynabeads were used in Clayton and co-workers survey ( Clayton et al. , 2001 ) . One disadvantage of this isolation method is that all the exosomes will merely be selected if they all contain the protein used for the isolation, otherwise, merely a proportion of the exosomes will be selected. A Good Fabrication Practice ( GMP ) approved method has been established for the isolation of exosomes used for clinical applications, which is based on extremist filtration and diafiltration followed by centrifugation on sucrose shock absorbers, ensuing in a extremely purified and unfertile exosome pellet ( Lamparski et al. , 2002 ) . This method is best suited for when large-scale exosomes isolations are needed and requires specialized equipment. There is a turn ing demand for a fast, efficient and dependable method that yields extremely purified exosomes suited for both research and clinical intents. Exosomes can do immune-activating responses in vitro Exosomes secreted from antigen-presenting cells contain a scope of immunostimulatory molecules that activate T cells, which suggest that they may hold an of import function in extension of immune responses.Exosomes derived from bacterially infected macrophages carry bacterial coat constituents and utilize these to excite a proinflammatory response in naif macrophages ( ONeil et al. , 2008 ) . It was demonstrated by Giri et Al that exosomes isolated from M. bovis and M. TB infected macrophages besides lead to activation and ripening of dendritic cells that derived from mouse bone marrow. Furthermore, intranasal disposal of mice with exosomes isolated from M. bovis BCG infected macrophages stimulated the coevals of memory CD4+ and CD8+ T cells. This indicated that the exosomes from infected macrophages might be a possible vaccinum campaigner for TB ( Giri et al. , 2008 ) .Human dendritic cell-derived exosomes ( Dex ) express NKG2D ligands ( activatory signals for Natural Killer cells ) such as MHC category I polypeptide-related sequence A ( MICA ) and MICB on their surface that bind to NKG2D that are present on NK cells and lead to NK cell activation ( Viaud et al. , 2009 ) . IL-2 and trans-presentation of IL-15 and IL-15 receptor a concatenation ( IL-15Ra ) is required for NK cell endurance, homeostasis and proliferation. The ability of Dex to advance NK cell proliferation was due to the coincident presence of IL-15Ra on these exosomes, which could adhere to exogenic IL-15 and let IFNg secernment by NK cells therefore stimulate NK cell proliferation.Dex express high degrees of functional MHC class-I and class-II peptide composites together with CD86 molecules and it has been suggested that Dex may replace for DCs to arouse MHC-class-I and class-II restricted T-cell responses and tumour rejection.Tumour-derived exosomes can besides hold immune-activating belongingss. This chiefly happens when the tumor cells are under stress conditions. The stress-inducible HSP 7 0 household is considered to work as an endogenous danger signal that can increase the immunogenicity of tumors and bring on cytotoxic T lymphocytes ( CTL ) responses. They are a cardinal portion of the cell s mechanism for protein folding, and they help to protect cells from emphasis.It was demonstrated that stimulation of 4T1 chest glandular cancer cells and K562 erythroleukemic cells with IFN-g leads to cellular emphasis response, which so triggers enhanced look of entire Hsp72 ( HSP70 household ; 72 kDa ) look without increasing cell programmed cell death significantly. It was revealed that Hsp72 is released within exosomes and has the ability to up-regulate CD83 look and excite IL-12 ( a T cell exciting factor ) release by naif dendritic cells ( Bausero et al. , 2005 ) .Under emphasis conditions, NKG2D ligands are up-regulated on tumour exosomes. Hsp70 besides activates mouse NK cells that recognize stress-inducible NKG2D ligands on tumor cells. In the SCID mice with Hsp70-over expressing tumors, NK cells were activated so that they killed ex vivo tumor cells that expressed NKG2D ligands ( Elsner et al. , 2007 ) Exosomes can besides hold immunosuppressive belongingss in vitro Immune equivocation by tumor cells is a good established mechanism wherein tumours avoid sensing and riddance by the host immune system. Tumour-derived membrane cysts have been reported to hold immunosuppressive maps for over 20 old ages ( Poutsiaka et al. , 1985 ) .Fas ligand ( besides known as CD95 ligand ) is a type II transmembrane protein of the Tumour Necrosis factor ( TNF ) household. When FAS ligand binds with its receptors, programmed cell death will be induced. Exosomes derived from tumor cell lines bear Fas ligand that can bring on T cell programmed cell death taking to suppression of T-cell responses in vitro. Addition of anti-FasL antibody blocked exosome-induced programmed cell death ( Abusamra et al. , 2005 ) .In malignant neoplastic disease patients, the loss of NKG2D, an energizing receptor for NK cells, CD8+ and NKT cells, is a critical mechanism of malignant neoplastic disease immune equivocation. NKG2D down-modulation is due to direct exosomal bringing of membrane -bound transmembrane growing factor b1 ( TGFb1 ) to CD8+ T cell or NK cell subsets ( Clayton et al. , 2008 ) . Down-regulation of NKG2D is partially due to the presence of the expressed NKG2D ligand MICA. Clayton and co-workers demonstrated that other possible NKG2D ligand such as MICB may be present on the surface of tumour-derived exosomes, and may lend to the down-regulation of NKG2D. On the other manus, some experiments added combinations of other NKG2D-ligand specific Abs did non accomplish a convincingly enhanced inhibitory consequence, but this may be consequence of the hapless barricading features of the Abs. In comparing, adding a TGFb1-blocking Ab about wholly abrogated the decrease in surface NKG2D. These informations suggest that down-regulation of NKG2D is chiefly due to the presence of TGFb1 and partially due to the presence of NKG2D ligands, but how exactly TGFb1 and NKG2D ligands cooperate requires elucidation.Interleukin 2 ( IL-2 ) is an of import cytokine which non merely supports enlargement and distinction of cytotoxic T cells and Natural Killer ( NK ) cells, but besides supports regulative T cells ( Treg cells ) and their suppressive maps ( Clayton et al. , 2007 ) . Tumour exosomes inhibit IL-2 induced T cell proliferation by advancing the map of Treg cells.It has besides been reported that tumour exosomes-mediated initiation of IL-6 plays a function in barricading bone marrow DC distinction. The degrees of IL-6 and phosphorylated Stat3 were elevated 12 hours after the tumor exosomes stimulation of murine myeloid precursors, and tumour exosomes were less efficient in suppressing distinction of bone marrow cells isolated from IL-6 smasher mice. It was suggested that bone marrow DC precursors capture tumour exosomes and later bring on the production of IL-6 and other cytokines, taking to the activation of Stat3. As a consequence, distinction of bone marrow precursors into immature DC is blocked ( Yu et al. , 2007 )Immune cell-derived cysts c an besides hold immunosppressive belongingss. Activated T cells secrete exosomes bearing FasL, which induces programmed cell death of bystander T cells, thereby take parting in activation-induced cell decease ( AICD ) . Ectosomes derived from neutrophils and erythrocytes inhibit the secernment of IL-8 and TNF by macrophages and the ripening of DCs ( Thery et al. , 2009 ) .Vesicles purified from some organic structure fluids can besides hold immunosuppressive activities. Exosomes from human chest milk in vitro inhibit T cell activation and increase the figure of Treg cells ( Admyre et al. , 2007 ) . It was suggested that exosomes present in chest milk may hold a function in the immune tolerance of the baby and may besides protect the baby from allergic reaction development.MIC molecules are reported to be constitutively transcribed in human placenta throughout normal gestation and soluble MIC molecules are released from in vitro cultured placenta explants.Elevated degrees of soluble MIC molecules are present in gestation sera and are able to down-regulate the NKG2D receptor and impair the cytotoxic map of peripheral blood mononucleate cells from healthy givers. Take together, these consequences suggest a fresh mechanism for immune equivocation of the fetal homograft through foetal MIC and maternal NKG2D interactions ( Mincheva-Nilsson et al. , 2006 ) . A more recent survey by the same group showed that the 2nd household of human NKG2D ligands, the UL-16 binding proteins ( ULBP ) , is besides expressed by placenta. Isolated placenta exosomes carried ULBP1-5 and MIC on their surface and induced down-regulation of the NKG2D receptor on NK, CD8+ , and gd T cells, taking to decrease of their in vitro cytotoxicity without impacting the perforin-mediated lytic tract ( Hedlund et al. , 2009 ) Antigen presentation by Exosomes Exosomes can show antigens straight or indirectly. Secreted membrane cysts carry both antigenic stuff and peptide-MHC composites. Exosomes can besides reassign antigens from tumour cells to DCs and maps in antigen cross-presentation. Exosomes purified from cultured tumor cell lines contain tumour antigens and can bring on the activation of antigen-specific T cells in vitro in the presence of recipient dendritic cells, which have otherwise non encountered the antigen.Secreted cysts display preformed peptide-MHC composites on their surface that can sometimes be straight presented to T cells. Exosomes and microvesicles derived from most cell types bear MHC category I molecules that could potentially bring on CD8+ T cell activation ( Thery et al, 2009 ) . It was reported that Dendritic cells ( DC ) derived exosomes could straight bring on the activation of CD8+ T cell in an antigen-specific mode ( Admyre et al, 2006 )Antigen showing cell ( APC ) derived exosomes express big sums of MHC c ategory II molecules. These exosomes can straight trip blood relation T cell ringers or pre-activated CD4+ T cells, but they need to reassign antigen and MHC molecules to dendritic cells, which will so trip naif CD4+ T cells ( Thery et al, 2009 ) . Shuttle for RNA A more recent progress in the field of exosome survey has been the find that these cysts can besides incorporate, and deliver, species of RNA molecules that can be active in cells that take up the exosomes. A survey by Skog et Al. showed that Glioblastoma-derived exosomes are enriched in messenger RNA and miRNA. They showed that messenger RNA could be delivered into receiver normal cells and bring forth a functional protein. It was besides shown that spongioblastoma microvesicles seem to excite proliferation of a glioma cell line ( Skog et al, 2008 ) . Valadi et Al. showed similar consequence, with a presentation that the messenger RNA nowadays in exosomes was active and functional ( Valadi et al. , 2007 ) . Their designation of RNA being present in exosomes supports the hypothesis that exosomes may be a bringing vehicle by which one cell communicates with another, really presenting RNA and, in bend, modulating recipient-cell protein production. Transportation of infective agent HIV atoms utilize multivesicular compartments that are enriched in MHC II and CD63 as the major site for accretion in human macrophages. And the virus released has proteins normally found on exosomes. These findings formed the background for the Trojan virus hypothesis , which suggested that viruses such as HIV could commandeer and conceal in exosomes secreted from an infected cell, and because the retroviruses are released in the signifier of exosomes, they could so get away the host defence. It has been demonstrated that the cellular prion protein ( PrPc ) and the transformed infective PrPscrapie ( PrPsc ) are found in exosomes secreted into the civilization medium and exosomes that express PrPsc were infective, bespeaking the function of exosomes in the spread of prions ( Fevrier et al. , 2005 ) The map of membrane secreted vesicles/exosomes in vivo The function of exosomes secreted in vivo is still in argument and demands to be to the full determined. Exosomes isolated from gestations later presenting at term expressed significantly higher degrees of biologically active constituents, including Fas ligand and HLA-DR, than those from gestations presenting preterm. A survey by Taylor et al indicated that exosomes with T-cell inhibiting activity are increased in pregnant adult females who deliver at full term comparison to adult females who deliver pre-term ( Taylor et al. , 2006 ) . Use of exosomes as possible immunotherapy vaccinum. Tumour cell-derived exosomes incorporating tumor antigens plus MHC category I molecules can reassign tumour antigens to DCs to bring on a CD8+ T cell dependent antitumour immune response. Exosomes released from DCs pulsed with tumor antigens were besides shown to arouse strong anti-tumour responses. One survey compared exosomes derived from tumor cells and exosomes derived dendritic cells in facet of the stimulatory efficiency of the antitumor immune responses induced by these two normally used exosome vaccinum. Dendritic Cell-Derived Exosomes Stimulate Stronger CD8+ CTL responses and antitumor unsusceptibility than tumour cell-derived exosomes ( Hao et al. , 2006 ) .Inoculation of CBA/J mice with exosomes secreted by SRDCs pulsed in vitro with Toxoplasma gondii-derived antigens ( TAg ) before gestation induced a protective response in the progeny. In fact, inoculation resulted in the presence of significantly fewer cysts in offspring encephalons ( Beauvillain et al. , 2009 ) . Clinical tests Two stage I clinical tests have been carried out on patients with advanced phase melanomas or non-small cell lung carcinomas showing melanoma-associated antigen ( MAGE ) . These tests required the constitution of good fabrication processs ( GMPs ) to obtain clinical-grade exosomes from patients DC loaded with tumour antigen derived peptides. These surveies proved that it is executable and safe to give exosomes to human topics and the exosomes are efficient in bring oning antigen-specific T-cell responses. The lone side effects were mild localised reactions at the site of injection and mild febrility in a few patients. The clinical results were encouraging, demoing transeunt stabilisation of the disease in half of the patients in the melanoma test and a 3rd of the patients in the lung carcinoma test.Another study that studied the usage of ascites-derived exosomes ( Aex ) in combination with the granulocyte-macrophages colony-stimulating factor ( GM-CSF ) in the immunotherapy of color ectal malignant neoplastic disease was besides published. 40 patients with advanced colorectal malignant neoplastic disease were involved in this survey and they were indiscriminately allocated to interventions with Aex entirely or Aex with GM-CSF. Both interventions were safe and the Aex plus GM-CSF can bring on tumour-specific antitumour cytotoxic T lymph cell ( CTL ) response ( Dai et al. , 2007 ) .Based on these consequences, a stage II clinical test is about to be initiated in France in patients with non-small cell lung malignant neoplastic disease that as been stabilized by chemotherapy. This survey will unite the disposal of exosomes from IFNg-treated DCs ( which express NKG2D ligands and IL-15Ra ) , and will use attacks that inhibit T Reg cells ( Mignot et al. , 2006 ) . Breast Cancer Breast malignant neoplastic disease is the 2nd major cause of malignant neoplastic disease decease in American adult females, with an estimated 40,170 lives lost in adult females in the United States in 2009 ( Jemal et al. , 2009 ) . Although ovarian malignant neoplastic disease histories for fewer deceases than chest malignant neoplastic disease, ovarian malignant neoplastic disease still represents 4 % of all female malignant neoplastic diseases. For some of the instances of both types of malignant neoplastic disease, there is besides a clear familial nexus.In 1994, two chest malignant neoplastic disease susceptibleness cistrons were identified: BRCA1 on chromosome 17 and BRCA2 on chromosome 13. When persons have a mutated BRCA1 or BRCA2 cistron, they are at an increased hazard of developing chest or ovarian malignant neoplastic disease at some phases of their lives. Until late, it was non clear what the map of these cistrons was, until surveies on a related protein in barm reveale d their normal function: they participate in mending radiation-induced interruptions in double-stranded DNA. It is thought that mutants in BRCA1 or BRCA2 might disenable this mechanism, taking to more mistakes in DNA reproduction and finally to cancerous growing.This will be the first survey that compares normal mammary epithelial cells-derived exosomes and breast cancer-derived exosomes. Research Proposal The primary purpose of my undertaking is to insulate and qualify exosomes and to compare the immune-modulating belongingss of exosomes secreted from normal chest epithelial cell line ( B42 ) and a tumour line derived from it ( B42 clone 16 ) ( Weaver et al. , 2009 ) . I will measure if the normal and malignant neoplastic disease derived exosomes influence lymphocyte maps and tumour growing. Both cell lines are cultured in a serum-free growing medium incorporating Clonetics Mammary Epithelial Basal Media ( MEBM ) and MEGM SingleQuot addendums ( Cambrex ) . This information will so be used in a comparative analysis of patient derived stuff.Flow cytometry was used at the start to place the belongingss of the atoms present in the cell civilization supernatant. PBS on its ain was compared to man-made beads of size 48 nanometer, 100 nanometer and 410 nanometer. And consequences show that the size of the little atoms present in the cell line supernatant was less than 100 nm as seen in 2F, 2 G and 2H. Analysis of the MEBM medium utilizing flow cytometry showed that there were little atoms with size less than about 300nm as seen in 2E. A fact that must be considered in all exosome surveies is that civilization media addendums, such as FBS, can besides incorporate exosomes. Therefore media supplememts should either be avoided if executable, or pre-spun at 100,000 ten g to take exosomes. To turn out that the little atoms we detected were secreted from the cells and non from the civilization medium, we so stained our cells with Carboxyfluorescein succinimidyl ester ( CFSE ) , which is a fluorescent cell staining dye. B42 cells were incubated with CFSE in PBS ( concentration of CFSE is 10 Â µM ) at room temperature in dark for 10 proceedingss. The CFSE was so removed by rinsing the cells and 10ml of growing medium was added in the flask, which was so put in the brooder. 1ml of supernatant was taken out after 24, 48 and 72 hours, and was so analysed by flow cytometry. P A ll atoms detected in supernatants from cells are fluorescent as seen in 3 A, 3B, 3C, 3D and 3E, which indicates that they are secreted from the cells.Immunoblotting has besides been utilised to characterize the stray cysts. 50ml of 5-day normal cell supernatant and 35ml of 5-day tumor cell supernatant was used. Exosomes were isolated utilizing centrifugation at 1500rpm ( 100 x g ) followed by ultracentrifugation at 24,000 revolutions per minute ( 100,000 x g ) utilizing a SW28 rotor for three hours. Supernatant was poured off and the pellets were resuspended in PBS. The protein concentration was determined to be: normal cell-derived exosome pellet 344 mg/ml, tumor cell derived pellet 535 mg/ml.Both samples were resuspended in SDS cut downing sample buffer ( 2 % SDS, 20 % Glycerol, 0.05 % bromophenol blue, 0.5 M Tris pH6.8 and Dithiothreitol as the cut downing agent ) and denatured by heating at 70 Â °C for 1 minute. Electrophoresis was carried out in SDS-PAGE running buffer for abo ut 1 hr at 150 VProtein samples were so transferred onto a nitrocellulose membrane ( in blotting buffer ( 25mM Tris, 192mM Glycine ) at 100 V for about 30 proceedingss ) . Ponceau S was used to look into the protein transportation. The nitrocellulose membrane was washed in H2O and so with PBS+ 0.1 % Tween 20. Barricading buffer ( PBS+0.1 % Tween and 5 % Skimmed milk pulverization ) was so added and the membrane was left on a shaker for about 15 proceedingss. Primary antibodies ( Alix, Tsg101. HC10 ) were diluted into PBS+ 0.1 % Tween ( 1/1000 dilution ) and incubated with the membrane and were left on a shaker in a cold room overnight.Membrane was washed 3 times with PBS+0.1 % Tween at 10 proceedingss intervals. Secondary antibody IgG-HRP was diluted in PBS+0.1 % Tween ( 1/1 0, 000 dilution ) and was incubated with the membrane for about 20 proceedingss. Membrane was so washed 3 times with PBS+0.1 % Tween at 10 proceedingss intervals. 1ml of PBS, 300 m cubic decimeter of Peroxide an d 300 m cubic decimeter of foil ( Super signal West Femto ) was assorted together and incubated with the membrane for about 1 minute. Excess fluid was so absorbed with tissue. Antibody binding was so visualised with Fujifilm Intelligent Dark Box LAS-3000.The consequences are shown in 6. Typical exosomes surface markers such as MHC Class I, ALIX and TSG101 can be visualised. As this is merely the preliminary consequence and the burden of normal and tumour exosomal protein was non equal, this experiment will be repeated in the hereafter. Future experiments: 1. It has already become evident that a confining factor in the experimental system we have developed is the sum of exosomes that can be harvested from civilization supernatants. Non-adherent lymphoid lines can be grown to high denseness, with good outputs of exosomes. In contrast, adherent cells, such as the chest lines being used in this survey, can non be grown to such high denseness in normal flasks, with a ensuing lower output of stuff. However, in communications with other exosome research workers, we are obtaining similar sums from these standard flasks.Therefore to better outputs we will look into the usage of fresh tissue civilization techniques. Two methods are executable. First, the usage of the Integra Celline system, which are particular flask incorporating a membrane edge inner compartment where cells are seeded, an external compa R tment where civilization medium is placed. This system is presently in usage by the Clayton group in Cardiff. Its chief disadvantage is the cost of each single flask is in the part of ?150. Whilst good pattern should let extended usage, accidental infection or the demand for a fresh batch of cells would necessitate another new flask. Therefore, the system will we prove will be the usage of microcarries, which are little dextran-based domains on which disciple cells grow. Adding these to standard flasks can therefore increase the surface country greatly.Cell growing and exosome production will be optimised utilizing Cytodex 1 bearer beads. A methodological analysis paper has been published for the usage of the Celline flask system in exosomes production, but non for microcarriers, therefore it is a possible paper as good.2. The flow cytometry informations shown above, whilst declarative mood of little cysts in the civilization medium, does non turn out that they are exosomes. We will utilize flow cytometric screening to farther characterise these cysts, with the end of identifying and screening microvesicles from exosom es. Such a technique could hold great value in the analysis of cyst biomarkers in patient samples.3. Exosomes will be obtained through standard high-velocity ultracentrifugation. The proteins present on these exosomes will be analysed in order to analyze the composing of the exosomes. Western smudge and flow cytometry will be used to analyze exosomes markers associated on the normal and tumour cells. two-dimensional gel electrophresis will be used to qualify any differences between normal and tumour cells, proteins can be so analysed by mass spectroscopy, which identifies the protein. Whether exosomes will impact the growing of blood lymph cells harvested from healthy givers will be studied. We will besides utilize tumour-derived exosomes to find if they affect the growing of Normal and tumour cells. Finally, we will analyze whether exosomes are present in the serum of chest malignant neoplastic disease patients and analyze its possible usage as a diagnostic tool.Experiments will be carried out to see whether tumour exosomes can intercede anchorage-independent growing of the chest malignant neoplastic disease cell line. We will besides analyze whether exosomes are present in the serum of chest malignant neoplastic disease patients. Mentions Abusamra, A, J. , Zhong, Z, H. , Zheng, X, F. , Li, M. , Ichim, T, E. ,Chin, J, L. A ; Min, W, P. ( 2005 ) . Tumor exosomes showing Fas ligand mediate CD8+ T-cell programmed cell death. Blood cells, Molecules, and Diseases. 35:169-173Admyre, C. , Grunewald, J. , Thyberg, J. , Gripenback, S. , Tornling, G. , Eklund, A. , Scheynius, A. A ; Gabrielsson, S. ( 2003 ) . Exosomes with major histocompatibility complex category II and co-stimulatory molecules are present in human BAL fluid. Eur Respir J. 22: 578-583.Admyre, C. , Johansson, S, M. , Paulie, S. A ; Gabrielsson, S. ( 2006 ) . Direct Exosome stimulation of Peripheral Human T Cells Detected by Elispot. Eur. J. Immunol. 36:1772–1781Admyre, C. , Johansson, S, M. , Qazi, K, R. , Filen, J, J. , Lahesmaa, R. , Norman, M. , Neve, E, P, A. A ; Scheynius, A. ( 2007 ) . Exosomes with Immune Modulatory Features Are Present in Human Breast Milk. J. Immunol. 179: 1969-1978Andre, F. , Schartz, N.E.C. , Movassagh, M. , Flament, C. , P autier, P. , Morice, P. , Pomel, C. , Lhomme, C. , Escudier, B. , Le Chevalier, T. , Tursz, T. , Amigorena, S. , Raposo, G. , Angevn, E. A ; Zitvogel, L. ( 2002 ) . Malignant gushs andimmunogenic tumour-derived exosomes. The Lancet. 360: 295-305.Bausero, M, A. , Gastpar, R. , Multhoff, G. A ; Asea, A. ( 2005 ) . Alternate mechanism by which IFN-g enhances Tumor acknowledgment: Active Release of Heat Shock Protein 72. J. Immunol. 175:2900-2912Beauvillain, C. , Juste, M, O. , Dion, s. , Pierre, J. A ; dimier-Poisson, I. ( 2009 ) . Exosomes are an effectual vaccinum against inborn toxoplasmosis in mice. Vaccine. 27:1750-1757Caby, M. P. , Lankar, D. , Vincendeau-Scherrer, C. , Rapo

The Doomed Journey of Panfilo de Narvaez in Florida

The Doomed Journey of Panfilo de Narvaez in Florida Panfilo de Narvaez (1470-1528) was born to an upper-class family in Vallenda, Spain. Although he was older than most Spaniards who sought their fortunes in the New World, he nevertheless was extremely active in the early conquest period. He was an important figure in the conquests of Jamaica and Cuba in the years between 1509 and 1512. He acquired a reputation for ruthlessness; Bartolome de Las Casas, who was a chaplain on the Cuba campaign, recounted horrible tales of massacres and chiefs being burned alive. In Pursuit of Cortes In 1518, the governor of Cuba, Diego Velazquez, had sent the young conquistador Hernan Cortes off to Mexico to begin the conquest of the mainland. Velazquez soon regretted his actions, however, and decided to place someone else in charge. He sent Narvaez, with a large force of more than 1,000 Spanish soldiers, to Mexico to take command of the expedition and send Cortes back to Cuba. Cortes, who was in the process of defeating the Aztec Empire, had to leave the recently subdued capital of Tenochtitlan to return to the coast to fight Narvaez. The Battle of Cempoala On May 28, 1520, the forces of the two conquistadores clashed at Cempoala, near present-day Veracruz, and Cortes won. Many of Narvaez’s soldiers deserted before and after the battle, joining Cortes. Narvaez himself was jailed in the port of Veracruz for the next two years, while Cortes retained control of the expedition and the vast wealth that came with it. A New Expedition Narvaez returned to Spain after being released. Convinced that there were more wealthy empires like the Aztecs to the north, he mounted an expedition that was doomed to become one of the most monumental failures in history. Narvaez got permission from King Charles V of Spain to mount an expedition into Florida. He set sail in April 1527 with five ships and about 600 Spanish soldiers and adventurers. Word of the riches earned by Cortes and his men made finding volunteers easy. In April 1528, the expedition landed in Florida, near present-day Tampa Bay. By then, many of the soldiers had deserted, and only about 300 men remained. Narvaez in Florida Narvaez and his men clumsily made their way inland, attacking every tribe they met. The expedition had brought insufficient supplies and survived by pillaging meager Native American storehouses, which caused violent retaliation. The conditions and lack of food caused many in the company to become ill, and within a few weeks, a third of the members of the expedition were severely incapacitated. The going was tough because Florida was then full of rivers, swamps, and forests. The Spanish were killed and picked off by irate natives, and Narvaez made a series of tactical blunders, including frequently dividing his forces and never seeking allies. The Mission Fails The men were dying, picked off individually and in small groups by native attacks. Supplies had run out, and the expedition had alienated every native tribe it had encountered. With no hope to establish any sort of settlement and with no help coming, Narvaez decided to abort the mission and return to Cuba. He had lost touch with his ships and ordered the construction of four large rafts. The Death of Panfilo de Narvaez It is not known for certain where and when Narvaez died. The last man to see Narvaez alive and tell of it was Alvar Nunez Cabeza de Vaca, a junior officer of the expedition. He recounted that in their final conversation, he asked Narvaez for help the men on Narvaezs raft were better fed and stronger than those with Cabeza de Vaca. Narvaez refused, basically saying â€Å"every man for himself,† according to Cabeza de Vaca. The rafts were wrecked in a storm and only 80 men survived the sinking of the rafts; Narvaez was not among them. The Aftermath of the Narvaez Expedition The first major incursion into present-day Florida was a complete fiasco. Of the 300 men who landed with Narvaez, only four ultimately survived. Among them was Cabeza de Vaca, the junior officer who had asked for help but received none. After his raft sunk, Cabeza de Vaca was enslaved by a local tribe for several years somewhere along the Gulf Coast. He managed to escape and meet up with three other survivors, and together the four of them returned overland to Mexico, arriving some eight years after the expedition landed in Florida. The animosity caused by the Narvaez expedition was such that it took the Spanish years to establish a settlement in Florida. Narvaez has gone down in history as one of the most ruthless yet incompetent conquistadors of the colonial era.

Economics of Global Money Markets Essay Example | Topics and Well Written Essays - 2750 words

Economics of Global Money Markets - Essay Example owered the discount rates and extended term loans and in the following month by decreasing the set targets for the rates of federal reserve by 50 points. As economic indications weaknesses advanced over successive months, the rates of federal funds target was lowered by 325 points by the committee, leaving it at 2% by the end of 2008 (Oulette, 2014). During the summer, the rates were held constant by the committee as it watched financial and economic conditions. As the crisis grew worse at the fall of summer, finance committee responded through lowering rates of federal funds target by 100 points, with this coming unparalleled cut of interest rate by central banks. In December, evidenced by dramatic slowdown, the committee lowered its target at a range of 0-25 points. Up to date the range as remained with no interference. In spite of monetary policy easing, under performance in credit markets worsened off. Towards the end of 2008 and at the start of 2009, the Fed took astonishing mea sures to give support and liquidity to functioning of credit markets inclusive of establishment of various emergencies loaning facilities and extension or creation of currency exchange agreements with fourteen central banks globally. As a regulator of all banks, the Fed led the largest America bank holding firms’ stress test, creating stage for firms to increase their capital (working mother, pp.66).The actions with interventions from policy makers in America and globally assisted to stabilize financial world markets, which later checked on the weakening of real economy with deflationary pressure emergence.Though it is probable that worse results had been stopped, the destruction to the economy was already severe. The rate of unemployment in the U.S rose from 6% in 2008 to 9% by 2009 as inflation decreased sharply. With the increase of the crisis, and rate of federal funds on the lower bound, the committee resorted to non-traditional boundaries approaches to counter the crisis. When

Comparison and contrast essay Anchorage,Alaska and Key West Florida

Comparison and contrast Anchorage,Alaska and Key West Florida - Essay Example Perhaps. Key West, Florida and Anchorage, Alaska may be thousands of miles apart geographically and culturally, but in some ways, they are practically next door neighbors. The first striking difference between these two cities is obviously geography and climate. Key West is the westernmost island of the archipelago known as The Florida Keys. Close proximity to the Tropic of Capricorn and the warm currents of the Caribbean Sea give Key West a balmy average temperature of 82oF. Add to this an annual rainfall of nearly 40 inches per year and you have a pleasant climate capable of supporting a host of interesting vegetation while providing many hours of sunny weather to enjoy the waters surrounding the island. Of course, any city that has taken on the title of â€Å"Southernmost City in America† had better have plenty of warm weather and sunshine to back up their claim. Anchorage, while not the northernmost city in America is a great deal different than Key West. One has no need to look beyond the raw numbers to see that two more different cities could not be found in America in regards to geography and climate. Anchorage boasts and average annual temperature of 35oF with an average annual precipitation of just 16 inches (most of it of the frozen variety)(Hoare, 2008). Though summer days can reach into the mid 70’s, summer and winter weather is unpredictable in Anchorage. 40oF swings in temperature have been known to occur within the space of several hours. Anchorage enjoys long summer days due to its high latitude, but the subarctic climate surrounding the city dissuades most people from spending those summer days swimming in the frigid coastal waters nearby. An additional geographic feature that makes Anchorage much different that Key West is the nearby presence of active volcanoes. Occasional ash accumulation from these volcanoes poses a slight

Park Tea Room Essay Example for Free

Park Tea Room Essay Master â€Å"Hally† Harold is the seventeen year old lead of Athol Fugard’s work, a white boy of South African descent, son of his mentor Sam’s employer. Sam is one of two black waiters employed by his family’s business, the St. George’s Park Tea Room. The focus of the play is of the two men’s mutual educating of the other. The younger of the two, Hally takes great pride in his â€Å"educating† Sam on book knowledge, the things that he has learned in reading or the classroom, whereas the elder Sam spends his days educating Hally on the ways of life and the world, showing him how important it is to take pride in oneself and the things that can be accomplished by your own hands. Hally has been caught in a dastardly position being a young man desperately in search of his place in this world as he rapidly approaches manhood, and being the only son of an immensely racist drunkard in the face of South African apartheid. Hally has found himself stuck between the ideologies of his inadequate father and the teachings of his gifted mentor. He battles himself for both loving and being ashamed of his white South African roots and alcoholic father. This play is about the corrosive power and denunciation of racism, ignorance and hatred in a society where those elements are all that surround you. Hally is a very bright young lad torn by his implied societal position and his loyalties to the man whom he feels has afforded him life’s greatest lessons, a black waiter who works for his father, Sam. Hally is a tortured and tormented soul; even the title of the play denotes the societal position of these individuals-Hally is referred to as Master Harold, a seventeen year old boy while Sam and Willie are grown men referred to as boys. The division between the races is clear, there is no â€Å"distortion of the political significance† (Jordan pp. 461) of the setting in which this work takes place, white is better than black and can in no socially significant way be mixed without ill regard. The only comfort to be found in the underlying premise of this work is the fact that Harold does not initially feed into the views of his father and society. He, in the beginning sees what great things he can learn from these black African men and chooses to err on the opposing side of his father’s views of race relations. It is not until Hally begins to feel trapped and cornered by his father’s impending release from the drunkard ward of the local hospital that he slips into the standard ideology of a white male finding his path during South African apartheid. He turns on his mentor, spits in his face and throws a total tantrum because he has not learned how to deal with all the scrapes and cuts that can come of being a man in this world. The introduction of Hally’s drunkard father back into the home is the unadulterated reason for his abruptly abusive and racist behavior toward â€Å"the boys†. His underlying fear is that he won’t be able to stand up for himself and his true beliefs if his father is present. Deep down I don’t feel that Hally believes himself to be any greater or more important than Sam or Willie but he is aware that society feels him to be superior to these two men and that he has yet to find it within him to give his own personal ideals a voice. He has spent all of his life under foot of one of the most racist men in South Africa, yet in the face of that socially and paternally enforced racism Hally has, for the most part, allowed himself to remain open-minded to the gifts and understandings of others, realizing that everyone has something to offer. His father’s hospital stay afforded Hally the time he needed to reflect on his own thoughts to determine what his outlook on this world would be. He was able to live without the weight of race long enough to become comfortable with himself as an individual and the other individuals surrounding him without regard to race or social standing. Being the intelligent lad that he is, he realizes that his father coming home means a lot for the way that he has been living his life, he is inevitably going to have to make some changes; he will either have to change the way that he views the world and begin fully subscribing to his father’s way of thinking, or he will have to find his own manhood and let his father know how he really feels. Hally is a clay chameleon being molded to fit whatever situation he finds himself in; he harbors an immense amount of disgust and disdain for his father and it is apparent at every turn except when he is speaking to his father. When engaging with the patriarch of his family Hally appears loving, caring and compassionate. He does not allow his hatred for his father’s world views to be seen by the man who gave him life, instead he hunts for the underlying love and respect that a son should have for his father as a man, and harnesses that love long enough to engage in an empathetic exchange. The fact that this young man has named the cycle of life the â€Å"principle of perpetual disappointment† speaks volumes of his outlook on the daily affairs of this world. He feels that having his father present in the home will just complicate the lives of everyone else around without justification; his father is just an impediment of unnecessary worth, a hurdle to be overcome if Hally ever desires to see himself find true happiness. As far as Hally is concerned, where reference is made to life being a dance as discussed in the play, it is his thought that no one knows the moves, no one man has all of the steps in order because no one can fully hear the music; as such the voluntary reality that these men discuss throughout the play could never exist. Just the thought of his father coming home changes Harold for the worse. Even in remembering the night that Sam strapped Harold’s father to his back and carried him home from the bar in the rain or the day that Sam took Harold under his wing and taught him not only to ‘fly a kite’ literally but symbolically by spreading his wings as a man and learning to fly on his own. The kite was merely a symbol to teach Harold how important it is to find his own way in this world, not to follow his father’s mind or anyone else’s other than his own. Yet where Sam felt that all these things made he and Harold closer, forging a bond that could not be broken, Hally instead turns on Sam stressing that he no longer refer to him as Hally but as Master Harold, signifying the social position and difference between the two. He does the one thing that Sam would have never expected him to do; he takes the position of the superior being and reduces Sam to a â€Å"nigger† thereby inflicting upon his former mentor an irreversible wound. Hally took his opportunity to put Sam in his place and let him know that no matter what Sam has been to him or done for him and his family over the years that he is not immune to the underlying hatred that erodes the human conscience in instances such as the time period in which this play has been set. Sam tries to make clear the implications of Harold’s actions and stresses the significance of what he has done to him, and their relationship, until the young lad comes to his senses and admits the effect that his love for his father has on him and his behaviors. Hally is fully dependent upon Sam for his understanding of this world because Hally can’t even understand himself. He lashes out at Sam because Sam is the closest person to him and sometimes it’s just easier to hurt the ones you love because you know better what will hurt them than a stranger, but I feel that another reason why he lashed out at Sam in such a way was because beneath it all he knew that Sam could always see his heart and his true intentions. Sam was able to discern and decipher the complex feelings that Hally had for his father and the emotions provoked by the idea of his father’s return. I understand the impressionable minds of youth but this young man is seventeen years old, it is time for him to stop relying on things like his relationship with Sam and to start making a way for himself. In a world full of adults you can’t just act out whenever you want to lashing out at those around you and always expecting people to be as forgiving as Sam was in this instance because it is my thought that the fact of the matter is-Hally was releasing some pinned up thoughts and emotions that he has been harboring, waiting for the day that he could release that portion of his father’s essence which he holds within him. There is no doubt that the relationship previously held between the two has forever been changed. Because he is seventeen years old the world says that it is time for this young lad to become a man, but he is not ready. He’s still relying upon others to tell him what he thinks and how he really feels. If he can’t handle the complexity of his thoughts and emotions for his father how could he ever hope to handle a life out in the world on his own. Harold knows that racism and hatred are wrong, both a lose thread eroding the fabric of life, but that makes no difference to him, when put in a position of discomfort he lashed out at Sam and Willie in the same manner that one would expect of a small child. In his article Boehmer makes it clear how often Fugard uses his main character to bring about the realization of conditions of separateness by shining a light on the trappings of historical pains, that his inevitable alienation has given representation to ordinary lives and not necessarily unique and therefore ‘dramatic’ situations† (Boehmer pp. 165). That is the point which commands emphasis in our analysis because there is nothing particularly special or significant about the setting of this play other than the backdrop of the apartheid era. Without knowledge of this story having taken place during the apartheid era these events could have taken place in any part of the world at any time throughout history. ‘Master Harold’ was no special case; he was a seventeen year old boy like any other seventeen year old boy enthralled in the decision to either follow in his fathers footsteps or to tread his own path. Cummings piece says that Fugards’ work â€Å"dramatizes the racial situation in South Africa† (Cummings pg. 2), this is true insofar as Fugard has taken the apartheid struggle and turned it into a dramatic work, as have many other artists, but not in such a way as for the thoughts or ideas of the characters within the play to have been exaggerated because just like I said, Hally was no special case. There was no need of exaggeration because we see young men like Hally everyday, unsure of themselves or their place in this world, worried that if they make a decision for their life that it may be the wrong one so they choose to sit idly in their comfort zone too afraid to venture into any unfamiliar territory. For Hally it would have been widely unfamiliar for him to stand up to his father and say, ‘thank you father, for giving me life, but my thoughts of this world should be formed of my own volition, not handed down from generation to generation’ and it is until just such young men can do that very thing that the older ideals of racism and hatred will begin to falter. Cummings is right about one thing though, the simplicity of the setting does largely contradict the complexity of the characters (Cummings pg. 2) but I think that it must be understood that if the setting and characters would otherwise be in constant competition with each other and no one would be able to follow the play. The characters are what carry the work. If Hally had no minutiae to set his character apart and was just another seventeen year old lead, there would be nothing pivotal to hold this play together. All the little details are what make these characters so profound and the work of such high quality; it would be a detriment to the production if anyone was to ever tamper with the formula. References Fugard, Athol. â€Å"Master Harold †¦ and the Boys†. New York: Penguin Plays (1982). Boehmer, Elleke. â€Å"Review: Speaking from the Periphery†. Third World Quarterly, Vol. 11, No. 1 (Jan. , 1989), pp. 161-166. Cummings, Mark. â€Å"Reclaiming the Canon: A World Without Collisions: â€Å" â€Å"Master Harold†Ã¢â‚¬ ¦and the Boys† in the Classroom†. The English Journal, Vol. 78, No. 6 (Oct. , 1989), pp. 71-73. Jordan, John O. â€Å"Life in the Theatre: Autobiography, Politics, and Romance in â€Å"Master Harold†Ã¢â‚¬ ¦and the Boys†. Twentieth Century Literature, Vol. 39, No. 4, Athol Fugard Issue (Winter, 1993), pp. 461-472. Solomon, Alisa. â€Å"Review: [untitled]-Reviewed work(s): †¦Master Harold†¦and the Boys by Athol Fugard†. Performing Arts Journal, Vol. 7, No. 1 (1983), pp. 78-83.

Aristotle’s Politics - The Good Man Should Not Rule the City Essay

Aristotle’s Politics - The Good Man Should Not Rule the City Aristotle contends that the good man is dissimilar to the good citizen in ways he goes a great length to illustrate. He distinguishes the two for the purpose of facilitating his later arguments concerning the appropriate allocation of sovereignty to the rightful ruler, who he subsequently claims is the good man who excels all others in each and every aspect. Aristotle's distinction further prompts the notion that he advocates a monarchial form of constitution, for the rule of a single good man is equivalent to a constitution of kingship. This can be derived through the following reasoning. Aristotle is convinced that the good citizen can so be defined only in relation to the constitution he is an element of: 'The excellence of the citizen must be an excellence relative to the constitution (1276b16).' The good man on the other hand, 'is a man so called in virtue of a single absolute excellence (1276b16).' He further asserts that the good citizen 'must possess the knowledge and capacit y requisite for ruling as well as for being ruledÃâ€"a good man will also need both (1277b7~1277b16).' From these conclusions of Aristotle, it is evident that the good man and the good citizen differ in the manner of their excellence, but not in their capacity for ruling or being ruled. It should therefore follow that there should not exist impediments to the ruling by the good citizen in the city as opposed to the ruling by the good man due to the fact that they are identical in their competence to rule. However, Aristotle in his later arguments, crowns the good man as ruler: 'in the best constitutionÃâ€"there is someone of outstanding excellence. What is to be done in that case? Nobody wou... ...scussed). The justification of the good man in becoming the supreme educator can be made in the following way. Since all absolutely excellent men (good men) arrive at their excellence through the process of education, that is, they are not innately excellent, their efforts should be directed toward the emulation of their excellence in the children of the city, for they are the ones who know best the process of becoming excellent. In this manner of education, the children (being future citizens) will grow up to become good men and good citizens, and thus the future city will comprise of many potential rulers. The good man through education, will contribute towards the ruling of the city indirectly in such an instance, and not directly as Aristotle claims he should do. Works Cited Aristotle. Poetics. Trans. Gerald F. Else. Ann Arbor: Ann Arbor Paperbacks, 1990.

Behavior is hereditary Essay

The whole issue of nature and nurture is complex with potential debates centered round their influences on human development. Nature refers to the traits inherited from parents while nurture refers to the influence of the environment to an individual’s behavior (Lippa 2005, p. 26). Psychological research shows that both nature and nature have a general influence on the physical and behavioral characteristics of people. This paper seeks to explain the factors which affect nature and nurture. The nature theory holds that human behavior is hereditary. True to the theory, some traits such as blood type and eye color are known to be predominantly genetic. It can therefore be asserted that genetic variation of people affects the development and characteristics of the next generation (Lippa 2005, p. 28). Nurture on the other side holds that the environment in which an individual is brought up in influences his or her behavior. In this context therefore, it can be acknowledged that the ability of an individual to interact and conform to the societal requirements and the environment at large affects the degree to which the environment can influence the individual’s behavior (Lippa 2005, p. 28). Still to be noted here is the effect of age on the influence of nature and nurture. Inherited traits are first noticed when a child is born. As the child grows up to teenage, behavioral characteristics are predominantly influenced by the environment because of the interactive nature of people at this stage of growth as well as the ability to conform to peers and the society at large (Lerner & Bearer 2004, p. 86). At adult stages, people tend to become more independent hence less influenced by the environment. At this stage therefore nature prevails but the nurture traits acquired at early stages of growth remains effective. In conclusion therefore, nature and nurture are undoubtedly influential to human development. Nature gives inborn traits while nurture takes these inborn abilities and changes them with age to fit the individual’s environment. ? Bibliography Lippa, R 2005, Gender, nature, and nurture, Lawrence Erlbaum Associates, Inc. Lerner, R, & Bearer, E 2004, Nature and Nurture: The Complex Interplay of Genetic and Environment. Routledge: New York.

My Summer at Choate Rosemary Hall

This summer at Choate Rosemary Hall was the best summer that I have ever had so far. The classes were challenging but also exiting at the same time. I really love the classes there because the number of student per one teacher is very small so the teachers give pay more attention toward you. Also, the student gets to know each other more so they can work together better. One of my favorite classes during the summer was Anatomy and physiology because we had a lot of hands on experience with dissection. My favorite dissection in the class is when we get to dissect and explore the anatomy and physiology of a rodent’s body because it was the longest and interesting dissection that we had during the summer program. When we finish with the dissection, our teacher takes a picture of our dissection and sends it to us and we had to label it. Another project that I like about Anatomy and Physiology is the owl pellet. We had to put back together a scattered rodent skeleton together and it was very time consuming. Other than the classes I also like the dorm life was. At home my parent do everything for me but at Choate I had to do everything myself, which made me feel like an adult. Every morning I had to wake up by myself and go to breakfast. My Favorite part about dorm life is laundry. My friend and I always wash our cloths together to save some money and we would just sit in the laundry room talking to each other about each other cultures. One time I forgot my highlighter in my pocket during washing so all of my cloths was ruin. The mistake of putting a highlighter in my pocket turns out really cool because it made one of my shirts really pretty so everyone said that I was the best mistake that I’ve ever made. Choate’s campus was big. On the first day of class I wanted to change my schedule and didn’t know anywhere to go to so I had to run around the campus trying to find the summer program office for almost an hour. Every day I have only 30 minutes for lunch because one of my classes was on the other side of the campus and it would take me 10 minutes to walk there. I think my favorite building at Choate is Hill House because that is my dorm, the dining hall, and it’s also the place that my friend and I hang out before study hours. Hill House has an air conditioned lounge that no one knows. Therefore, it was our secret hideout and our hanging spot. Also every Saturday we held a movie night at Hill House because checks in is at 11 p. m. My favorite part about this summer is spending time with friends that I had made during the program. In the beginning I didn’t know anybody else other than the Memphis Prep scholars so I stayed with them. After the first week I start to know more people so I hang out with other people. We would go to the movie almost every Wednesday because we had nothing to do. Sometime we would go to the library to read mangas and books because some of us had to finish our summer reading list. Usually after we go to the library we would go to the candy shop down the street from the library to get ice-cream. Sometime we would skip dinner at Choate’s dining hall and have dinner at a restaurant that is next to the candy shop. The one thing that I hate the most about this summer program is saying good bye. On the last week of the program we eat dinner together at a restaurant every night or ordering Chinese food, trying to spend as much time with each other as possible. Also, on the last mall trip we would try to buy things to gift each other so that we will remember each other. On the night of the Last Dance we went the basement and talked to each other and signed each other shirts and journals. The last day of the summer program was the hardest day for me because all of my friends were leaving. We cry every time someone is leaving and we would stand there watch them leave and by the end of the day all of my tears were dried up. Attending Choate’s summer program is the best thing that had happened to me. The classes and teachers were great. The campus was big and it made me feel like I’m in college. I had made a lot of new friends that I will never forget. Choate summer program is also life changing for me because I learn how to be independent from my parents and do things on my own. It had expanded my knowledge in Anatomy and Physiology. I also learn that there are a lot of opportunities out there waiting for you to take it and you have to work hard to achieve it. The best opportunity that I had taken this year is to apply for Memphis PREP because without them I wouldn’t know about Choate Rosemary Hall or any other boarding school.

Life and Work of Maud Lewis, Canadian Folk Artist

Life and Work of Maud Lewis, Canadian Folk Artist Maud Lewis (March 7, 1903 – July 30,  1970) was a 20th-century Canadian folk artist. With a focus on subjects in nature and ordinary life and a folk style of painting, she became one of the best-known artists in Canadian history. Fast Facts: Maud Lewis Occupation: Painter and folk artistBorn: March 7, 1903 in South Ohio, Nova Scotia, CanadaDied: July 30, 1970 in Digby, Nova Scotia, CanadaParents: John and Agnes DowleySpouse: Everett LewisKey Accomplishments: Despite physical limitations and poverty, Lewis became a beloved folk artist, known for her brightly colored paintings of animals, flowers, and outdoor scenes.Quote:  Ã¢â‚¬Å"I paint all from memory, I don’t copy much. Because I don’t go nowhere, I just make my own designs up.† Early Life Born Maud Kathleen Dowley in South Ohio,  Nova Scotia, Lewis was the only daughter of John and Agnes Dowley. She had one brother, Charles, who was older than her. Even as a child, she suffered from rheumatoid arthritis, which limited her movements, even down to her hands. Despite this, she began making art at an early age under the tutelage of her mother, who taught her to paint watercolor Christmas cards, which she then sold. Maud dealt with multiple physical disabilities that left her hunched over. At the age of fourteen, she dropped out of school for unknown reasons, although it is possible that the bullying of her classmates (due to her visible birth defects) was at least partially at fault. Family and Marriage As a young woman, Maud became romantically involved with a man named Emery Allen, but they never married. In 1928, however, she gave birth to their daughter, Catherine. Allen abandoned Maud and their daughter, and they instead continued to live with her parents. Because Maud had no income and no means to support her child, a court required Catherine to be placed up for adoption. Later in life, an adult Catherine (now married with a family of her own and still living in Nova Scotia) attempted to get in touch with her mother; she was never successful in her attempts. Maud’s parents died within two years of each other: her father in 1935 and her mother in 1937. Her brother Charles inherited everything, and while he allowed his sister to live with him for a short while, she soon moved to Digby, Nova Scotia, to live with her aunt. In late 1937, Maud answered an advertisement placed by Everett Lewis, a fish peddler from Marshalltown, who was seeking a live-in housekeeper. While she was unable to perform her job well, due to the advancement of her arthritis, Maud and Everett married in January 1938. Painting Every Surface The painted interior of Maud Lewis home, as it is preserved in the Art Gallery of Nova Scotia.   Courtesy of the Art Gallery of Nova Scotia. The Lewises lived mostly in poverty, but Everett did encourage his wife’s painting – especially once he realized they could make a small profit. He procured  painting supplies  for her, and she then accompanied him on selling trips, starting with small cards like those she had painted as a child and eventually expanding to other, larger media. She even painted nearly every suitable surface in their small home, from typical sites such as walls to more unconventional ones (including their stove). Because canvas was difficult to come by (and expensive), Maud worked on beaver boards (made of compressed wood fibers) and Masonite, among other things. These smaller items, early in her career or for personal use, were full of bright colors and designs of flowers, birds, and leaves. This aesthetic would carry over into her later work as well. Early Sales Maud Lewis,  White Cat (2), 1960s, oil on pulpboard, 31.1 x 33.8 cm. Collection of the Art Gallery of Nova Scotia, gift of Johanna Hickey, Vancouver, BC, 2006.   Maud’s paintings, throughout her career, focused on scenes and items out of her own life, experiences, and surroundings. Animals appeared frequently, mostly domestic or farm animals such as cows, oxen, cats, and birds. She also portrayed outdoor scenes: boats on the water, winter sleigh or skating scenes, and similar moments of ordinary life, often with a playful and cheerful tone. The greeting cards of her youth came back again, this time as inspiration for her later paintings. Bright, pure colors are a hallmark of her paintings; in fact, she was known to never blend colors, but only use the oils as they came originally in their tubes. Most of her paintings are quite small, not exceeding eight by ten inches. This is mostly due to the constraints of her arthritis: she could only paint as far as she could move her arms, which was increasingly limited. However, there are a few of her paintings that are larger than that, and she was commissioned to paint a large set of shutters by American cottage owners in the early 1940s. Gaining Wider Attention Maud Lewis,  Fall Scene with Deer,  c. 1950, oil on pulpboard, 29.5 x 34.9 cm. Collection of the Art Gallery of Nova Scotia, purchase 1974. During her lifetime, Maud’s paintings did not sell for large amounts. By the late 1940s, tourists had begun to stop at the Lewises’ home to purchase her paintings, but they rarely sold for more than a few dollars. In fact, they wouldn’t sell for even close to ten dollars until the final years of her life. The Lewises continued to live a meager existence, with Everett taking on the lion’s share of work around the house as Maud’s arthritis continued to degenerate her mobility. Despite the attention of the occasional tourist, Lewis’s work remained fairly obscure for the majority of her life. All that changed in 1964, when the Toronto-based national newspaper  Star Weekly  wrote an article about her as a folk artist and brought her to the attention of audience across Canada, who quickly embraced her and her work. The attention only increased the following year, when the broadcasting network CBC featured her on its program  Telescope, which featured Canadians of varying degrees of notoriety who had made a difference in some way. In the final years of her life and following these major public mentions, Lewis was on the receiving end of commissions from a wide array of important figures – most notably, American president  Richard Nixon  commissioned a pair of paintings from her. She never left her home in Nova Scotia and was unable to keep up with the demand for artwork. Death and Legacy Maud Lewis,  Maud Lewis House, mixed media, 4.1 x 3.8 m. Collection of the Art Gallery of Nova Scotia, purchased by the Province of Nova Scotia, 1984.   Maud’s health continued to deteriorate, and in the late 1960s, she spent most of her shuttling between painting in her home and visiting the hospital for treatment. Her declining health was exacerbated by the wood smoke of their home and the constant exposure to paint fumes without proper ventilation, and the lung issues this caused left her susceptible to pneumonia. She died on July 30, 1970, after battling pneumonia. After her death, demand for her paintings skyrocketed, as did the appearance of forgeries. Several paintings purported to be Maud’s were eventually proven to be fakes; many are suspected to be the handiwork of her husband Everett in an attempt to continue cashing in on her prominence. In recent years, Maud’s paintings have only grown more valuable. She has become something of a folk hero in her home province of Nova Scotia, which has long embraced artists with authenticity and unusual styles, and in Canada as a whole. In the 21st  century, her paintings have sold at prices well into five figures. After Everett’s death in 1979, the Lewises’ house began to fall into disrepair. In 1984, it was purchased by the Province of Nova Scotia, and the Art Gallery of Nova Scotia took over the care and preservation of the house. It now dwells in the gallery as part of a permanent exhibit of Maud’s works. Her paintings have made her a folk hero among the Canadian art community, and the bright joyfulness of her style, combined with the humble, often harsh realities of her life, have resonated with patrons and fans worldwide. Sources Bergman, Brian. â€Å"Paying Tribute To Painter Maud Lewis.†Ã‚  The Canadian Encyclopedia, https://www.thecanadianencyclopedia.ca/en/article/paying-tribute-to-painter-maud-lewis/Stamberg, Susan. â€Å"Home Is Where The Art Is: The Unlikely Story of Folk Artist Maud Lewis.†Ã‚  NPR, https://www.npr.org/2017/06/19/532816482/home-is-where-the-art-is-the-unlikely-story-of-folk-artist-maud-lewisWoolaver, Lance.  The Illuminated Life of Maud Lewis. Halifax: Nimbus Publishing, 1995.

Spanish Words Without a One-Word English Equivalent

Spanish Words Without a One-Word English Equivalent Here are some Spanish words for which there is no good one-word English equivalent: Amigovio/amigovia This word is fairly new and has various meanings dependent on context, but it often refers to a live-in girlfriend or boyfriend, or a friend with benefits. Katrina era mi amigovia, pero desde que estoy con Belà ©n no la vi ms. (Katrina was my friend with benefits, but since Ive been with Belà ©n I dont see her any more.) Anteayer The day before yesterday, formed by combining ante (before) and ayer (yesterday). Ese es el restaurante al que fuimos anteayer. (That is the restaurant we went to the day before yesterday.) Aprovechar Although this verb can often be translated simply as to use, it often carries a strong connotation of getting the most out of something. No pude aprovechar de la piscina del hotel porque prefiero la playa. (I couldnt take advantage of the hotels swimming pool because I prefer the beach.) Bimestre As a noun, a bimestre is a period of two months. El ahorro en un bimestre fue de 2,500 euros. (There was a savings of 2,500 euros in two months.) As an adjective, bimestre means bimonthly (every two months). Cacerolazo A type of protest in which participants bang on pots and pans. Los cacerolazos comenzaron ayer temprano en Buenos Aires. (The pots-and-pans protest began early yesterday in Buenos Aires.) Centenar A group of 100. The word is often used as an approximation for a large group. Un centenar de inmigrantes de origen subsahariano han intentado saltar la valla de Melilla. (About 100 sub-Saharan immigrants have tried to jump the fence at Melilla.) Concuado/concuada Someone married to a sibling of your spouse is a concuà ±ado or concuà ±ada. La esposa del hermano de mi esposa is mi concuà ±ada. My wifes brothers wife is my concuà ±ada. Consuegro/consuegra If you have a child, his or her spouses parents are your consuegros. For example, your sons wifes mother would be your consuegra. No sà © si mi consuegra es mi amiga or mi enemiga. (I dont know if my childs spouses mother is friend or foe.) Decena Just as a docena is a dozen or group of 12, a decena is a group of 10. Puedes comprar flores por decena. (You can buy flowers 10 at a time.) Desvelarse This reflexive verb refers to not getting sufficient sleep. Cuando nos desvelamos, el cuerpo obtiene energà ­a de una fuente ms accesible: la comida. (When we dont get enough sleep, the body gets energy from the most accessible source: food.) Someone who is sleep-deprived is desvelado. Empalagar To be excessively sweet, either literally (as of some candy) or figuratively (as with personality). Te voy a enseà ±ar mi receta particular, por causa de que la original me empalaga. (Im going to send you my own recipe, because the original is too sweet for me.) Entrecejo The area above the nose, below the forehead and between the eyebrows. Quiero quitarme sin dolor los pelos del entrecejo. (I want to painlessly remove the hairs between my eyebrows.) Estrenar To wear, use, perform or display something for the first time. Estrenà © las zapatillas que mi marido me habà ­a regalado. (For the first time, I put on the slippers that my husband had given me.) A noun form, estreno, can refer to a movie or play premiere or similar events. Friolento Sensitive to cold. Era friolento y se cubrà ­a los hombros con un suà ©ter. (She was sensitive to the cold and covered her shoulders with a sweater.) Friolero has the same meaning. Estadounidense From or of the United States. American is a common equivalent but is sometimes ambiguous as it can also refer to someone or something from the Americas. La cultura estadounidense est construida por la clase media. (U.S. culture was built by the middle class.) Internauta Internet user. Internauta is not capitalized and can be masculine or feminine. El acceso a Internet para la poblacià ³n mexicana representa al menos 55.3 millones de internautas. (Internet access for the Mexican population is available to at least 55.3 million Internet users.) Manco Having one arm, either from birth or as an amputee. Un dà ­a conocà ­ una muchacha manca. (One day I met a one-armed girl.) Quincena This is a period of 15 days or two weeks. British English has fortnight, which is seldom used in American English. Esta quincena es la ms tranquila del aà ±o en Pamplona. (This two-week period is the quietest of the year in Pamplona.) The word can also refer to the 15th of the month, when many workers in some countries get paid. Sobremesa After-meal conversation, from sobre (over) and mesa (table). Con poco tiempo para la sobremesa volvemos a la carretera. (With little time to chat after eating, we went back to the highway.) Trasnochar To stay up all night or put in an all-nighter. Nos trasnochbamos jugando juegos como Islander y Super Mario. (We would stay up all night playing games such as Islander and Super Mario.) Tuerto Having only one eye, or seeing with only one eye. Se llama Pirata por ser tuerto. (He was called Pirate because he had one eye.) Tutear To speak to someone using tà º, the familiar form of you. A cultural equivalent might be to speak with someone on a first-name basis. Nunca habà ­a llegado al punto de tutear a alguien. (I never have reached the point of using tà º with someone.)

Criminal Research, Final Exam Research Paper Example | Topics and Well Written Essays - 250 words

Criminal , Final Exam - Research Paper Example The author says that the problems will persist in the younger generation if not addressed immediately. 9. The author can use the results from this case study and compare them to results after a youth treatment program; he will be able to see the extent to which the treatment program has helped the youth. 10. The effect due to the involvement of the national institute of justice will have; some of the youths have spent time in jail for crimes committed and may not feel free to attend any function that is promoted by the institute. 15. Qualitative data example is the information that the analysts of the case study attained from the study that is what causes the high rates of violence; Qualitative data can be used to show the extent of a problem. Two results from survey can be compared by looking the intensity and the effect of the problem. 20. Snowball sampling is a technique for developing research samples where the existing subjects recruit future subjects from among their acquaintances. It can be used to analyze for crime causes in dangerous areas where a victim of an attack names a friend or person who has undergone the same and can be interviewed. 21. A focus group is a qualitative method of research, here the group is asked questions on believes, opinions and perceptions towards a product, concept, service or an idea. A focus group is a good method of research when if comes to matters that affect the society at large e.g. in the study of drug abuse. 22. A researcher may have the problem of time constraints for open ended questions and will finally not get the best answer and results from the interview. It may be difficult for a researcher to get the respondents full attention. 23. Going native refers to case where the researcher stops acting as a researcher and becomes a full member of the group, participating fully in the group events. The problem with this approach is that