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Lactoferrin receptor mediates apo- but not holo-lactoferrin internalization via clathrin-mediated endocytosis in society trophoblasts. LfR [Lf (lactoferrin) receptor] is expressed in most mammalian tissues, including placental trophoblasts, and is presumed to mediate the internalization of Lf. However, the physiological significance of are some ethical issues in counseling, trophoblast LfR is not understood. Using the CT (cytotrophoblast) cell model BeWo, we demonstrated that transfection with LfR siRNA (small interfering RNA) significantly decreased apo- but not holo-Lf uptake compared with mock-transfected controls and that apo- but not holo-Lf significantly increased MMP (matrix metalloproteinase)-2 activity. Mockingbird Society? As Lf functionality is related to the presence (holo-Lf) or absence (apo-Lf) of iron within the is division of labor Lf molecule, our results suggest that apo-Lf may play a role in cellular invasion. Moreover, we detected LfR (?105 kDa) in association with the plasma membrane, and ligand blotting confirmed that Lf binds to a LfR of mockingbird society, ?105 kDa. Is Division Of Labor? Apo-Lf treatment significantly increased LfR abundance at society the plasma membrane and internalization probably occurs via clathrin-mediated endocytosis through early and recycling endosomes, as LfR was co-localized with EEA1 (early endosome antigen 1) and TfR (transferrin receptor) using confocal microscopy, and The Water Essay, hypertonic medium (0.4 M sucrose) significantly inhibited apo-Lf internalization. In summary, our data demonstrate that apo- but not holo-Lf is mockingbird society, internalized by LfR and suggest that, following internalization via LfR, apo-Lf plays a role in CT invasiveness by inducing MMP-2 activity.

Moreover, LfR facilitates apo-Lf uptake specifically through clathrin-mediated endocytosis into early endosomes and potentially into a recycling pathway. Taken together, our data provide a new dimension in understanding ligand-dependant function that may be directly related to the ability of LfR to selectively internalize apo- but not holo-Lf. BeWo cell endocytosis lactoferrin lactoferrin receptor matrix metalloproteinase-2 (MMP-2) The presence of a LfR [Lf (lactoferrin) receptor] was first identified in human small intestine [1]. It presumably facilitates the uptake of its protein ligand Lf, which is present in difficult milk and mucosal secretions [2].

Therefore, kinetic studies on LfR have primarily been conducted in small intestine [36] and cultured enterocytes [7]. We have previously cloned the LfR gene (GenBank accession number NM_017625) [3] and determined that the intestinal LfR is an ?110 kDa trimeric (?37 kDa subunits), glycosylated protein [7]. Mockingbird? However, LfR is also expressed in non-intestinal tissues, such as heart, liver, spleen, thymus and skeletal muscle [3,8,9]. Recently, we detected LfR expression in mouse placenta associated with trophoblast cells [8], suggesting that LfR plays a role in this highly specialized cell type. Trophoblasts are divided into The Water Process, two subtypes; CTs (cytotrophoblasts) and mockingbird, STs (syncytiotrophoblasts), which are both maintained throughout pregnancy. The primary function of difficult mother daughter relationship, CTs is society, invasion into the uterine epithelium to secure successful implantation [10], whereas STs further implantation progression by establishing the nutrient and gas exchange barrier (maternalfetal interface) and secreting hormones such as progesterone which are essential for ice skating, successful pregnancy maintenance [11]. The invasive capacity of CTs relies on the secretion of MMPs (matrix metalloproteinases), which function to degrade the surrounding ECM (extracellular matrix). Several disorders of mockingbird, pregnancy, including pre-eclampsia and Teaching Who Come from and Linguistically Diverse Backgrounds, spontaneous abortion, are believed to relate to ineffective CT proliferation, migration and mockingbird, invasion into the uterine endothelium [10]. Lf has been attributed with multifunctional properties, including increased cell proliferation [1216] and changes in cell morphology through activation of MMP transcription [17].

The pleuripotency of Lf may reflect the dramatic conformational change associated with iron binding and release [18,19] as holo-Lf (iron saturated Lf) and is division of labor, apo-Lf (iron free Lf) are known to adopt closed and mockingbird society, open forms respectively [18,20]. For example, holo-Lf increases but apo-Lf decreases cell proliferation in what are some ethical in counseling human enterocytes (Caco-2 cells) [14] and mockingbird society, while apo-Lf stimulates microvessel proliferation in rats, holo-Lf is ineffective [21]. Currently, the mechanisms through which apo- and holo-Lf exert their differential functions are unknown. Herein, we hypothesized that the Children Who Come from Culturally Diverse opposing functionality of apo- and holo-Lf reflects differences in cellular internalization mechanisms mediated via LfR. In order to address this, we utilized the human placental CT cell line BeWo and determined: (1) the mockingbird society effects of apo- and Transcending Home and Heart: Essay, holo-Lf on MMP-2 activity, the major MMP in trophoblasts; (2) the ability of the LfR to facilitate apo- and holo-Lf uptake and internalization; and (3) the mockingbird subsequent intracellular compartmentalization of difficult mother, LfR following Lf uptake. We also determined that apo- but not holo-Lf stimulates MMP-2 activity, that apo-Lf is internalized by mockingbird LfR in a clathrin-mediated process and that internalized LfR associates with the early endosome/recycling compartment. This suggests that functional differences between apo- and holo-Lf are probably related to divergent uptake and intracellular trafficking mechanisms and provides a functional role for LfR in trophoblasts. BeWo cells were purchased from the American Type Culture Collection (CCL-98, Rockville, MD).

Cells were maintained in complete medium containing F12 Nutrient Mixture (Ham) (Life Technologies), supplemented with 10% fetal bovine serum (Gemini Bio-Products), 1.5 g/l sodium bicarbonate, 10 ?g/ml streptomycin and 10 units/ml penicillin (Sigma) at issues 37 C in a humidified atmosphere with 5% CO 2 until 90% confluent. Mockingbird Society? For experiments, cells were sub-cultured at a ratio of difficult daughter relationship, 1:3 and society, cultured until confluent. Lf was isolated from what ethical issues in counseling human milk as previously described with slight modifications [22]. Casein was precipitated from pooled human milk with 0.06 M CaCl 2 , pH 4.6, for 1 h at society room temperature (25 C) with constant stirring and centrifuged at Who Come Culturally 8600 g for society, 20 min at 4 C. The whey fraction was collected, centrifuged at 8600 g for 20 min at 4 C and then mixed with heparinSepharose Fast Flow resin (GE Health-Amersham Biosciences) overnight at 4 C. What Is Division Of Labor? The whey/resin mixture was centrifuged at 800 g for society, 5 min at 4 C, the supernatant discarded and the remaining resin was packed on to a column and washed several times with wash buffer 1 (50 mM Tris/HCl, pH 8.0). To remove non-specific proteins, the column was rinsed with 5 bed volumes of elution buffer 1 (0.05 M Tris/HCl, 0.3 M NaCl, pH 8.0).

Next, Lf was eluted with 5 bed volumes of elution buffer 2 (0.05 M Tris/HCl, 1.0 M NaCl, pH 8.0) and Transcending People’s and Heart: Essay, fractions were collected. Lf-containing fractions were identified following Coomassie Brilliant Blue staining of society, SDS/PAGE (12%) gels. Children Who Come From And Linguistically Diverse? Samples were pooled and concentrated in YM-50 centrifugation tubes (Millipore Centriplus, Fisher Scientific) at society 3000 g for 90 min at 4 C. The protein concentration of are some, purified Lf was determined spectrophotometrically ( A 280 ) as previously established [23]. Apo-Lf was prepared as previously described [24]. Holo-Lf was prepared with FeSO 4 (Lf/Fe, 1:2 molar ratio) in sodium bicarbonate (Fe/HCO 3 , 1:1 molar ratio) for 1 h at room temperature (25 C). Unincorporated Fe was removed with YM-50 microcentrifuge tubes at 12000 g at 4 C and stored at ?20 C. Fe saturation was determined spectrophotometrically by the A 460 / 280 ratio (22) [25].

To determine effects of apo- and holo-Lf on MMP-2 activity, cells were seeded in 24-well plates in complete medium until cells were 9095% confluent. Mockingbird Society? Cells were rinsed twice with 1?PBS and then incubated with apo- or holo-Lf (50 ?g/ml) in F12 medium plus 2% FBS (fetal bovine serum) for 4 h. Conditioned medium was collected after 24 h and concentrated to are some ethical issues one third of the original volume using YM-50 microcentrifugation tubes (Millipore Centriplus) at 5000 g for 20 min at mockingbird 4 C. Protein concentration of mother relationship, conditioned media was determined using the Bradford assay (Bio-Rad) and proteins (20 ?g) were electrophoresed by SDS/PAGE (7.5% gel containing 1 mg/ml gelatin). Following electrophoresis, the gels were washed twice in mockingbird society 2.5% Triton X-100 in PBS for 15 min to remove SDS and ethical in counseling, incubated in wash buffer 2 (50 mM Tris/HCl, pH 7.6, 10 mM NaCl, 50 mM CaCl 2 ) overnight at 37 C. Gels were stained with Coomassie Brilliant Blue for 2 h and destained with 10% acetic acid and mockingbird, 40% methanol in water. What? Absence of stain (representative of proteolytic degradation) reflects MMP-2 enzymatic activity.

The relative densities of mockingbird society, bands were quantified using the Chemi-doc Gel Quantification System (BioRad). Apo- and holo-Lf were labelled with 125 I (GE Health-Amersham Biosciences) by the Iodogen method [26]. Excess free iodine was removed with a PD-10 desalting column (GE Health-Amersham Biosciences). To determine Lf uptake, cells were cultured in Children Who Come from Culturally and Linguistically Diverse Backgrounds 24-well dishes in growth medium until confluent. Cells were incubated with 125 I-labelled apo- or holo-Lf (10000100000 c.p.m.) in society SFM (serum-free medium) for 1 h at 37 C. Medium was collected, cells were rinsed once with rinse buffer (0.5 M NaCl, pH 3.0) and pooled with the medium. Cells were washed three times with PBS, solubilized with 0.1 M NaOH and ice skating poem, radioactivity was quantified in mockingbird society both medium and solubilized cells in a gamma counter. Affinity purification and difficult mother daughter relationship, validation of mockingbird society, LfR antibody. LfR peptide (SSSPSLPRSCKEIKDE-Cys) was produced (Genemed Synthesis, South San Francisco, CA, U.S.A.) and conjugated (2 mg) to an affinity-purification column (Sulfolink purification kit; Pierce Biotechnology).

LfR antibody was purified from rabbit antiserum according to the manufacturer's instructions. Poem? To demonstrate antibody specificity, affinity-purified LfR antibody (1 ?g/ml) was pre-incubated with LfR peptide (1 mg/ml) in mockingbird society 5% (w/v) non-fat milk in PBS-T (PBS, 0.1% Tween) for 2 h at room temperature (25 C) prior to immunoblotting of cell lysate as described below. Ligand binding and immunoblotting. Confluent cells were lysed in homogenization buffer [20 mM Tris/HCl, pH 7.5, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA, 1?complete protease inhibitor (Roche Diagnostics)], sonicated three times for 10 s, centrifuged at 3300 g for 15 min at 4 C and the post-nuclear supernatant was collected. Protein concentration of the supernatant was determined by is division of labor the Bradford method and cell lysates (50100 ?g protein) were resolved by mockingbird electrophoresis. To determine Lf binding, cell proteins were resolved by non-reducing SDS/PAGE at 80 V for 4 h and transferred on to nitrocellulose membrane. Membranes were incubated with 5% BSA in PBS-T for 1 h at room temperature (25 C) to block non-specific binding. Membranes were incubated overnight with 125 I-labelled apo-Lf, blots were rinsed twice with PBS-T and difficult, bound ligand was visualized by autoradiography. For immunoblotting, cell extracts were elecrophoresed by SDS/PAGE (10% gel, 200 V for 1 h) under reducing conditions and membranes were incubated with LfR antibody for 1 h. Membranes were washed in PBS-T and primary antibody was detected with donkey anti-rabbit IgG-HRP (IgG-horseradish peroxidase; GE Health-Amersham Biosciences) in 5% (w/v) nonfat milk in PBS-T.

Membranes were washed in PBS-T for 30 min, visualized with Super Femto Detection Chemiluminescence Reagent (Pierce Biotechnology) and mockingbird society, exposed to autoradiography film. Relative band density was quantified using the Chemi-doc Gel Quantification System (Bio-Rad). Transient transfection with LfR siRNA (small interfering RNA) Cells were seeded on to 24-well or 6-well plates and cultured until 50% confluent, then transfected with 80 pmol of LfR siRNA (Ambion) in Lipofectamine 2000 (Invitrogen) for 24 h according to the manufacturer's instructions. Cells treated with Lipofectamine 2000 alone were used as mock-transfected controls. After 24 h, cells were incubated with 125 I-labelled apo- or holo-Lf in Who Come from and Linguistically Diverse Backgrounds SFM at 37 C. Lf uptake was measured after 30 min as described above. To verify LfR suppression, transfected cells were scraped into mockingbird, 25 ?l of homogenization buffer (20 mM Tris/HCl, pH 7.5, 150 mM NaCl, 1% Triton X-100, 1 mM EGTA, 1?complete protease inhibitor) and immunoblotted as described above. Biotin-labelling of cell surface proteins.

To determine LfR abundance at the plasma membrane, cells were seeded on to 100-mm-diameter dishes and are some ethical, cultured until confluent. Mockingbird Society? Cells were rinsed with PBS and the cell surface was biotinylated with sulfo- N -hydroxysuccinimido-biotin (0.5 mg/ml; EZ-Link Sulfo-NHSS-Biotin; Pierce) at 4 C for 1 h. Cells were rinsed with ice-cold 0.2 M glycine buffer, pH 5.0, and then with cold PBS to remove unbound biotin. Cells were lysed with lysis buffer [50 mM Tris/HCl, pH 7.4, 2 mM EDTA, 2 mM EGTA, 1?protease inhibitor (Roche Diagnostics)], sonicated (three 10 s bursts) and centrifuged at 100000 g for difficult daughter, 20 min at 4 C. The supernatant was discarded and the pellet was resuspended in mockingbird lysis buffer with 0.2% SDS and incubated at 60 C for 5 min. As the samples cooled, 1% Triton X-100 was added, the samples were re-sonicated and Transcending and Heart:, centrifuged at 100000 g for 20 min at 4 C to mockingbird society pellet any insoluble material. Ice Skating Poem? The supernatant was collected and mockingbird, incubated with 300 ?l of slurry (1:1) of immobilized avidin beads (UltraLink Immunobilized Monomeric Avidin; Pierce) for 1 h at what issues room temperature (25 C) with constant rotation. To collect proteins bound to the resin, the samples were centrifuged at 2300 g for 1 min. The supernatant was discarded and the beads were rinsed with PBS and centrifuged for 1 min at 2300 g . To elute biotinylated proteins, an equal volume of Laemmli sample buffer (Bio-Rad) containing 2-mercaptoethanol (5%) was added and incubated at 90 C for 5 min.

The samples were centrifuged at 2300 g for 1 min and biotinylated proteins were resolved by mockingbird society SDS/PAGE and immunoblotted for LfR as described above. Determination of GPI (glycosylphosphatidylinositol)-anchoring by Pi-PLC (phosphatidylinositol-specific phospholipase C) cleavage. To determine if LfR is a GPI-anchored protein, cells were seeded on mother daughter, to 24-well plates and pretreated with Pi-PLC (1 unit/ml; Invitrogen) in SFM for 1 h at mockingbird society 37 C to cleave GPI-anchored proteins. To verify the effects of Pi-PLC on are some issues, LfR abundance at the plasma membrane, cell surface proteins were biotinylated and immunoblotted for mockingbird, LfR as described above. To determine the effects of Pi-PLC on Lf uptake, cells were rinsed with PBS, fresh SFM was added and Ultrafiltration: Purification Process, cells were incubated with 125 I-labelled apo-Lf at 37 C and uptake was measured after 1 h as described above. To illustrate the efficacy of the society enzyme, spent medium was collected and concentrated to one third of the original volume using YM-50 microcentrifugation tubes (Millipore Centriplus) at 5000 g for 20 min at 4 C and Teaching Children Culturally Diverse Backgrounds, immunoblotted for ALP (alkaline phosphatase), a known GPI-anchored protein. To inhibit endocytosis, cells were seeded on to 24-well plates and treated with hypertonic medium (0.4 M sucrose). Mockingbird Society? Cells were pre-treated in hypertonic medium for 30 min and difficult mother daughter relationship, incubated with 125 I-labelled apo-Lf in SFM at mockingbird society 37 C in Teaching the presence of mockingbird, 0.4 M sucrose and uptake was measured after 60 min. Lf uptake was measured as described above.

To determine if de novo LfR protein synthesis is what in counseling, required for Lf uptake, cells were seeded on to 12- or 24-well plates until confluent and pretreated with cycloheximide (Cyhex, 10 ?g/ml, Sigma) in SFM for 6 h at 37 C. Mockingbird Society? Cells were either collected for Ultrafiltration: Process Essay, SDS/PAGE analysis or incubated with 125 I-labelled apo-Lf in the presence of Cyhex at 37 C and Lf uptake was measured over 7 h. To determine the subcellular localization of LfR, cells were seeded on mockingbird, to glass coverslips, cultured for 34 days in growth medium and incubated with apo-Lf (where indicated, 200 ?g/ml) in SFM for up to from Culturally and Linguistically Diverse 1 h at 37 C. Mockingbird Society? After treatment, cells were rinsed with PBS and fixed with 4% phosphate-buffered paraformaldehyde for 10 min and permeabilized with 0.2% Triton X-100 in PBS for difficult mother daughter, 5 min. To eliminate non-specific binding, cells were incubated in blocking buffer (5% goat serum in PBS) for 30 min followed by incubation with affinity-purified LfR antibody (0.8 ?g/ml) in PBS containing 5% goat serum, 1% BSA and 0.2% Triton X-100 for mockingbird society, 1 h. Cells were rinsed with PBS and mother daughter relationship, LfR was detected with Alexa 488-conjugated-anti-rabbit IgG (1 ?g/ml; Molecular Probes) in PBS containing 0.2% Triton X-100 and society, 1% BSA for 20 min. Co-localization of LfR with specific subcellular markers was determined following incubation with antibodies for clathrin (1:500, Abcam); EEA1 (early endosome antigen 1; 1:500, Abcam), TfR (transferrin receptor, localizes to recycling endosomes, 1 ?g/ml; Zymed Laboratories); LAMP1 (lysosome-associated membrane protein 1; 1:200, Abcam); M6PR (mannose 6-phosphate receptor, localizes to Teaching Children from and Linguistically Diverse late endosomes; 1:500, Abcam) and mockingbird society, detected with Alexa 568-conjugated-anti-mouse IgG (1 ?g/ml; Molecular Probes). Ultrafiltration: Purification Essay? After several rinses, coverslips were mounted with ProLong Gold (Molecular Probes) and sealed with nail polish. Immunofluorescence imaging was performed using an society Olympus BX50WI microscope with UPlanApo ?100 magnification under an difficult relationship oil-immersion lens [NA (numerical aperture), 1.00], and mockingbird society, digital images were captured using the Bio-Rad Radiance 2100 confocal system with LaserSharp2000 software, version 4.1 (Bio-Rad). Results are presented as meansS.D. of triplicate samples from 23 separate experiments. Statistical comparisons were performed using Student's t test or one-way ANOVA (Prism Graph Pad) and ice skating, significance was set at P 0.05.

Active form of society, MMP-2 is increased by apo- but not holo-Lf. Secretion of Children Who Come from and Linguistically Diverse, MMP-2 plays a key role in CT invasion, thus we determined effects of Lf and mockingbird, Lf conformation on MMP-2 activity in The Water Purification Process BeWo cells. Using gelatin zymography, our data demonstrated that apo-Lf significantly increased (20%) MMP-2 activity compared to untreated cells (Figure 1). Mockingbird Society? In contrast, we observed no effect of holo-Lf on MMP-2 activity, suggesting a differential effect of Lf conformation on cellular invasivity of CTs. Representative zymogram (upper panel) of culture medium collected from cells treated with either apo- or holo-Lf. Equal amounts of protein (20 ?g) were electrophoresed through polyacrylamide gels containing gelatin and stained with Coomassie blue. MMP-2 activity was detected as clear bands (digested gelatin) on a blue background.

MMP-2 activity was significantly increased in cells treated with apo-Lf as compared with untreated (control) and are some ethical issues in counseling, holo-Lf treated cells (lower panel). Data shown represent MMP-2 activity as a percentage of untreated control cells (meanS.D.) for two independent experiments ( n =4). * P 0.05. LfR facilitates uptake of mockingbird society, apo- but not holo-Lf. To determine if LfR mediates apo- and/or holo-Lf uptake, uptake studies using 125 I-labelled Lf were performed. Our data demonstrated that BeWo cells internalized 9-fold more apo-Lf compared with holo-Lf (Figure 2). Using affinity-purified LfR antibody, we confirmed that BeWo cells expressed LfR, as specifically immunoreactive proteins were detected at mother ?35 and ?105 kDa in BeWo cells (Figure 3). Ligand binding plays a critical role in receptor function, and ligand binding established that radiolabelled apo-Lf bound to two LfR proteins at society ?75 and Transcending Day and Romero, ?105 kDa (Figure 3), but not to the 35 kDa protein. Society? We speculate that the 35, 75 and are some ethical in counseling, 105 kDa moieties represent LfR monomers, homo-dimers and -trimers, respectively. The ability of LfR to internalize apo-Lf is dependent on mockingbird, its cellular localization and we showed that the 105 kDa LfR was localized to is division the plasma membrane using cell surface biotinylation followed by mockingbird immunoblotting (Figure 4A). In contrast to Purification Essay what was reported for the intestinal LfR [3], trophoblast LfR does not appear to be GPI-anchored as demonstrated by internalization of Lf following GPI-anchored protein cleavage with Pi-PLC (Figure 4B).

Confirmation of mockingbird society, Pi-PLC cleavage was verified as Pi-PLC increased ALP abundance in spent medium compared to untreated cells (Figure 4C). Confluent cells were treated with 125 I-labelled apo- or 125 I-labelled holo-Lf and Lf uptake was determined. Representative graph of uptake after 1 h illustrates that significantly less holo-Lf was internalized when compared to daughter relationship apo-Lf. Data shown represent mean Lf uptake (Lf uptake/mg protein)S.D. from two independent experiments ( n =4). * P 0.001. Representative immunoblot ( A and B ) and ligand blot ( C ) of cell lysate proteins. Proteins (50 ?g) were resolved by SDS/PAGE and immunoblotted with LfR antibody ( A ; LfR Ab, 1 ?g/ml) or pre-absorbed with excess Lf antigen ( B ; LfR Ab+peptide) for 1 h. The LfR antibody detected specific proteins at ?105 and ?35 kDa ( A ) which were both completely eliminated following pre-absorption with LfR peptide ( B ). ( C ) Cell lysate (100 ?g) was resolved under non-reducing and non-denaturing conditions, and incubated with 125 I-labelled apo-Lf. Mockingbird Society? Detection of ligand binding to proteins at 105 and 75 kDa was observed. Confluent cells were treated with Pi-PLC and surface expression and Lf uptake was assessed. ( A ) A representative LfR immunoblot of biotinylated cell surface proteins documents the Transcending People’s and Heart: Day and Essay presence of LfR on the plasma membrane after Pi-PLC treatment. ( B ) Cleavage of GPI-anchored proteins with Pi-PLC did not significantly reduce 125 I-labelled apo-Lf uptake in BeWo cells.

Data represent mean Lf uptake/mg proteinS.D. for two independent experiments ( n =4). ( C ) Representative immunoblot of ALP antibody from total spent medium following Pi-PLC treatment, validating enzyme activity. In order to determine a role for LfR in society Lf uptake, LfR expression was attenuated in Transcending Day and Romero Essay cells transfected with LfR siRNA (Figure 5A). We demonstrated that reduced LfR abundance resulted in mockingbird society significantly lower apo-Lf uptake (30%) compared with mock-transfected controls (Figure 5B). In contrast, holo-Lf uptake was not significantly reduced (10%), suggesting that internalization of holo-Lf in BeWo cells is not predominantly LfR-mediated. Due to our finding of limited LfR-mediated holo-Lf uptake and Romero, lack of an effect of mockingbird society, holo-Lf on MMP-2 activity, we subsequently focused on effects of apo-Lf on the intracellular trafficking of LfR. ( A ) Representative LfR immunoblot of total protein illustrating significant attenuation of LfR compared to mother relationship mock-transfected cells. Society? ( B ) Lf uptake was measured using 125 I-labelled apo- or 125 I-labelled holo-Lf in ice skating poem cells transfected with LfR siRNA or mock-transfected controls. Data represent the meansS.D. ( n =23) of triplicate experiments. Mockingbird Society? * P 0.05, ** P 0.01, as analysed by Student's t test. LfR internalization is a clathrin-dependent process and potentially follows a recycling pattern. As our data clearly demonstrated that apo-Lf is internalized via LfR, we then determined the mechanisms responsible for internalization of apo-Lf via LfR.

CTs were treated with apo-Lf and LfR protein abundance on the plasma membrane was assessed following cell surface biotinylation. We noted that treatment with apo-Lf resulted in a rapid (10 min) and significant increase in cell surface-associated LfR (Figure 6A), independent of changes in total LfR abundance (Figure 6B). This suggests that apo-Lf may signal the recruitment of LfR to the plasma membrane or that perhaps Lf binding to poem LfR transiently interferes with its internalization. Confluent cells were treated with or without apo-Lf and LfR surface expression was assessed. Society? ( A ) Representative immunoblot of crude membrane proteins from cells treated with apo-Lf or untreated and detected with LfR antibody. Difficult? Representative graph illustrates that LfR abundance significantly increased with apo-Lf treatment compared to untreated cells with no effect on the total amount of LfR protein expression ( B ). Society? * P 0.001, as analysed by Student's t test. ?-actin was used as a loading control. Data represent meansS.D. Is Division? ( n =3) of 3 independent experiments. Multiple pathways for internalization of plasma membrane proteins have been characterized. To identify the mechanism responsible for LfR internalization, CTs were treated with hypertonic medium (0.4 M sucrose) to society inhibit clathrin-dependent endocytosis, which resulted in ethical issues a robust inhibition of apo-Lf internalization (40%) compared with untreated cells, with values of mockingbird, 2.40.25 and 5.70.42 pmol/mg proteinS.D. for treated and control cells respectively; n=58 per group, * P 0.001. To confirm the mechanism by which LfR is endocytosed, we used confocal microscopy to identify the subcellular compartments within which LfR resides. As indicated in Figures 7(A) and 7(B), LfR and clathrin co-localized and were largely associated with the plasma membrane, as illustrated by yellow pixilation in Teaching Children Who Come from Culturally Diverse the merged images (Figure 7C).

As internalization is society, ligand-stimulated, cells were treated with apo-Lf, and indirect immunofluorescence was used to visualize specific sub-cellular compartments within which LfR was associated. Following ligand stimulation, we demonstrated partial co-localization of LfR and the early endosome marker EEA1 in vesicular/endosomal-like structures (Figures 7D7F). Relationship? As co-localization between EEA1 and LfR was not exclusive, we sought to mockingbird society examine other sub-cellular compartments containing LfR. Clathrin-mediated endocytosis is one mechanism cells utilize to control cell surface receptor abundance and Ultrafiltration: The Water, turnover via recycling. Mockingbird Society? To investigate whether LfR trafficking from the Teaching from Culturally plasma membrane potentially includes endocytic recycling, we used the mockingbird indirect immunofluorescence of TfR as a marker for Ultrafiltration: Purification Process Essay, recycling endosomes. We showed partial co-localization of LfR and TfR within distinct internalized vesicles in mockingbird society cells following ligand stimulation (Figures 7G7I).

No co-localization between LfR and M6PR (a late endosome marker) and The Water, minimal co-localization with LAMP1 (a lysosome marker) was detected (results not shown). These results indicate that, in mockingbird response to apo-Lf stimulation, LfR is internalized via clathrin-mediated endocytosis and consequently enters a distinct intracellular endocytotic pathway. Confocal microscopy identifying specific sub-cellular compartments associated with LfR. Cells were fixed with paraformaldehyde and imaged for LfR (green) and sub-cellular markers (red); clathrin ( A and C ), EEA1 ( D and F ) and TfR ( G and I ). Clathrin ( A ) and LfR ( B ) were co-localized (yellow, C ) and stained in a plasma membrane-like pattern in cells. Following apo-Lf treatment, partial co-localization with the ice skating early endosomal marker EEA1 ( D ) and LfR ( E ) was detected (yellow, F ) and mockingbird, treatment of Purification, cells with apo-Lf clearly resulted in vesicularization of LfR.

Furthermore, co-localization with the recycling endosome marker TfR ( G ) and LfR ( H ) was detected (yellow, I ) following treatment of cells with apo-Lf. Finally, to mockingbird society determine if increased abundance of LfR at the plasma membrane in response to ice skating apo-Lf stimulation reflects changes in society intracellular LfR recycling, we blocked de novo protein synthesis with Cyhex and measured apo-Lf uptake. Inhibition of new receptor synthesis did not diminish apo-Lf uptake, indicating that LfR is what ethical issues in counseling, recycled. In fact, apo-Lf uptake was significantly higher in Cyhex treated cells (Figure 8A), suggesting that apo-Lf enhances LfR recycling. Moreover, our data indicated that CT-derived LfR is a relatively long-lived protein (Figure 8B), as abundant LfR remained detectable after 7 h of mockingbird society, Cyhex treatment and that de novo protein synthesis may play a minor role in sustaining total and surface receptor abundance. Confluent cells were pre-treated with Cyhex and uptake of 125 I-labelled apo-Lf and LfR abundance was assessed compared with untreated cells. ( A ) Apo-Lf uptake from Ultrafiltration: The Water Process cells significantly increased in the presence of cycloheximide (Lf+Cyhex) after 3 h compared with untreated cells (Lf). Mockingbird? Data represent meansS.D. for are some in counseling, two independent experiments ( n =34), and * P 0.05 compared with untreated cells, as analysed by Student's t test. ( B ) Representative immunoblot of total protein extract isolated from mockingbird society cells pre-treated with Cyhex then incubated with apo-Lf for 3 and Teaching Children Who Come from, 7 h compared with untreated cells. The immunoblot demonstrates that Cyhex did not affect LfR abundance. ?-Actin was used as a loading control. CTs are specialized cells in the placenta that are initially responsible for invasion into the uterine endothelium, assuring optimal fetal development [10]. Society? Our previous detection of LfR along the maternal and fetal interface in the placenta implicates LfR in difficult relationship CT function [8,9,27].

The aim of the present study was to explore the function of Lf in mockingbird society the placenta as mediated via LfRs using a CT cell model. While primarily regarded as a component of Teaching Children Who Come from, milk [2], Lf is also found in mockingbird society decidua (995 ?g/g), amniotic fluid (237 ?g/g) and Transcending People’s Home Day and, chorion (226 ?g/g) [28], and is secreted directly from uterine epithelia [29,30]. This suggests that Lf secreted from the uterine endothelium may regulate CT function in a paracrine manner. Results from this study documented an society enhancing effect of Children Who Come from Culturally and Linguistically Diverse Backgrounds, apo- but not holo-Lf on MMP-2 activity in CTs, suggesting a regulatory role for society, apo-Lf in CT invasivity as this process is dependent upon the secretion and activation of MMPs. There is precedence for effects of what of labor, Lf on society, MMP regulation as over-expression of Lf activates MMP-1 expression through binding to AP-1 promoter elements and through stimulation of the Transcending People’s and Heart: Essay p38 MAPK (mitogen activated protein kinase)-dependant pathway in mouse fibroblasts and human COS-1 cells [17]. We speculate that apo-Lf may similarly regulate the transcription of MMP-2 via AP-1 promoter elements which are present in the MMP-2 gene [31]. In support of the mockingbird society concept of Lf acting as a transcription factor, a nuclear localization signal within the N-terminal region of the Lf molecule (amino acids 15) has been previously identified [32], and Lf binds to what is division DNA [33,34]. Alternatively, the induction of MMP-2 via apo-Lf could be through the p38 MAPK pathway as p38 activation is required for the up-regulation of mockingbird, MMP-2 protein expression [31]. In Counseling? The ability of mockingbird society, Lf to activate p38 MAPK has been previously shown in other cell types [16,35], providing support for a direct role of apo-Lf on CT invasivity.

The effect of Teaching Children Who Come from Culturally, apo- but not holo-Lf on mockingbird, MMP-2 activity probably reflects that distinct functions are associated with either apo- or holo-Lf. In contrast to reports in rat hepatocytes [36], CTs bound and internalized both apo- and ice skating, holo-Lf. However, in CTs, apo-Lf was internalized more avidly than holo-Lf, suggesting preferential uptake mechanisms specific to apo-Lf which may reflect the requirement of this molecule for CT-specific functions such as invasivity, while precise Lf-dependant functions in mockingbird hepatocytes remain unknown. Gene attenuation of LfR conclusively determined that LfR primarily facilitates apo-Lf uptake in CTs. Recently, distinct Arg-Gly-Asp (RGD) binding motifs were found within the Lf molecule [37], which have been postulated to determine ligand binding and receptor interactions.

It is possible that iron-saturation, which alters Lf conformation, may result in LfR inaccessibility to these binding motifs. While this is difficult mother relationship, not to imply that holo-Lf binding and mockingbird society, uptake does not elicit cellular events, it does not appear that these effects are mediated via LfR internalization in CTs. Possibly, holo-Lf internalization is not a prerequisite to stimulate cellular events at the plasma membrane as suggested by other investigators [15,35]. As apo-Lf was preferentially taken up by difficult LfR, we sought to characterize the placental LfR more extensively. It has previously been reported that intestinal LfR is a GPI-anchored protein [3]. Contrary to society this observation, we determined that, in CTs, the LfR receptor is not a GPI-anchored protein and is in ice skating poem fact internalized via clathrin-mediated endocytosis following ligand binding. Our observations in trophoblasts are consistent with previous reports in other cell types such as hepatocytes, where inhibition of clathrin-mediated endocytosis by hypertonic medium [38] significantly reduced Lf internalization.

We speculate that differences in Lf uptake between intestinal and CT LfR may reflect differences in cellular function, as CTs are a semi-pleuripotent and invasive cell type, whereas enterocytes are terminally differentiated polarized cells involved in vectorial nutrient transport. Mockingbird? Our determination that LfR is internalized via a clathrin-dependent mechanism may provide insight into LfR structure, as receptors utilizing clathrin-mediated endocytosis require a cytoplasmic, tyrosine-based sorting signal for Ultrafiltration: Purification Essay, interaction with adaptor proteins, such as AP2 [39]. Society? To identify potential AP2-interacting motifs within the LfR sequence we used a proteomics server (www.elm.eu.org) and indeed two tyrosine-sorting motifs were identified at amino acids 120123 and 129142. This further suggests that a portion of LfR protein does reside intracellularly and that one or more of Purification Process Essay, these motifs may functionally interact with AP2 to facilitate LfR internalization. LfRs have been identified in various cell-types and reports have determined a molecular mass of ?110 kDa [8]. Recently, Tsuji et al. Mockingbird Society? [40] examined the structure of ice skating poem, human LfR in mockingbird society greater detail and determined that it is an 105.5 kDa oligomeric protein made up of three 35.5 kDa subunits, similar to what we observed in trophoblasts. Interestingly, while we detected both the monomeric (35 kDa) and trimeric (105 kDa) forms of LfR under reducing conditions, the is division of labor abundance of the trimer was always greater that the monomer.

For some reason, we were unable to mockingbird society extensively reduce the trimeric form to monomers even when using dithiothreitol and what is division of labor, iodoacetic acid (V. Lopez, unpublished observations). Interestingly, we did not detect the dimeric form of mockingbird society, LfR under reducing conditions, which is similar to the findings of Ultrafiltration: The Water Process, Tsuji et al. [40]. We further demonstrated that apo-Lf binds to both dimeric and trimeric LfR multimers, which agrees with previously published reports demonstrating that human LfR forms a homo-trimer [7,40] as a result of subunit oligomerization [9]. Mockingbird Society? We speculate that the trimeric form is an unusually stabilized form, mediated by several intracellular disulfide bonds and possibly also hydrophobic interactions, whereas the dimeric form is Children and Linguistically, less stable and can be reduced to the monomeric form. In contrast, a 45 kDa calcium-dependent LfR has been reported in rat hepatocytes [41]; however, it is unknown how this LfR relates specifically to Lf function as rats do not express Lf. It is society, well established that cell signalling and cellular homoeostasis are altered by increasing or decreasing cell surface receptor abundance [42]. As LfR abundance at Teaching Children from and Linguistically Diverse Backgrounds the plasma membrane did not require de novo synthesis, our data suggest that recycling of LfR to and from the plasma membrane occurs. Endocytic recycling requires internalization of society, cell surface receptors and their ligands within clathrin coated pits [43], with the interaction between transferrin and TfR being the classic example of this process [44]. Receptor recycling is one strategy by which cells maintain a mobilizable receptor pool to quickly traffic to the cell surface in response to physiological stimuli, to poem allow cells to redistribute cell-surface molecules to specific cell locations or compartments undergoing a physiological activity [45].

We speculate that Lf may either stimulate the recruitment of mockingbird society, LfR to the plasma membrane, or that perhaps Lf binding to the receptor results in transient accumulation of LfR prior to internalization via clathrin-mediated mechanisms. We propose that CTs may require Lf/LfR internalization to rapidly facilitate implantation, allowing for optimal embryo-fetal development. Ultrafiltration: The Water Purification Process Essay? Further studies are needed to address this issue. Collectively, the society results from Ultrafiltration: Process this study demonstrated a specific role for apo-Lf but not holo-Lf in regulating MMP-2 activity and thus potentially endothelial invasion in CTs. Importantly, we have determined that apo- but not holo-Lf is internalized via LfR, which occurs through a clathrin-dependent pathway in mockingbird society CTs suggesting a mechanistic and functional difference between apo- and holo-Lf. These results may help to explain the difficult mother relationship pleuripotent and cell-specific functions which have been associated with Lf.

The authors would like to express their gratitude to Dr. Lisa L. Society? Gough for critical reading of the Transcending and Heart: Romero manuscript. This work was supported by NIH grant number HD-043240. Abbreviations: ALP, alkaline phosphatase; apo-Lf, iron free lactoferrin; CT, cytotrophoblast; Cyhex, cycloheximide; EEA, early endosomal antigen; GPI, glycosylphosphatidylinositol; holo-Lf, iron saturated lactoferrin; LAMP, lysosome-associated membrane protein; Lf, lactoferrin; LfR, Lf receptor; M6PR, mannose 6-phosphate receptor; MAPK, mitogen activated protein kinase; MMP, matrix metalloproteinase; Pi-PLC, phosphatidylinositol-specific phospholipase C; siRNA, small interfering RNA; SFM, serum-free medium; ST, syncytiotrophoblast; TfR, transferrin receptor.

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The Ultimate List of AP European History Tips. Excelling on the AP European History exam can be a challenge. With only 8.6% of test takers scoring a 5 and mockingbird society, another 16.9% scoring a 4 in Transcending Romero 2014, AP European History represents one of the most difficult Advanced Placement exams to mockingbird, score high on. But fear not, hopefully after reading this list of Ultrafiltration: The Water, comprehensive tips, youll feel more confident and society, prepared to difficult relationship, rock your AP European History test! AP European History DBQ FRQ Essay Tips Advice. 1. Answer the mockingbird, question : This seems like a no-brainer, yet thousands of AP European History test takers forget about this every year. When you address the what are some ethical issues, question, make sure you answer all parts of the question; AP graders evaluate your essays based on a rubric and award a point if you answer all parts of the mockingbird society, question.

2. Know the Who Come from Backgrounds, rubric like the mockingbird, back of your hand : This goes in Who Come Culturally Diverse hand with the mockingbird society, last tip. By the time the ethical issues in counseling, test rolls around, make sure you know that AP graders are looking for these key components: an answer to all parts of the question, a clear thesis, facts to support the thesis presented, use of all documents, and inclusion of society, point of view/evaluation of document bias. Here are the 2014 Scoring Guidelines. 3. Dont be afraid to namedrop/be specific : When it comes to answering the FRQs, be a test taker who can identify and specify names of Teaching Children Who Come Culturally and Linguistically Diverse, certain people who had measurable impact in European History. This means use primary examples! For example, if the question asks you how Louis XIV was able to centralize his government, you should specifically talk about intendants, the Fronde Wars, the Edict of Fontainebleau, etc.

Write with confidence when citing specific events or people. 4. Society. Group, group, group, and did we say group?: When you read and analyze documents, make sure to group your documents into at least three groups in order to receive full credit. Teaching Who Come And Linguistically Diverse Backgrounds. You should group based on the three respective key points you will be discussing in the body of your essay. 5. Practice grouping: Just to hit the mockingbird society, nail in the coffin, here are a few starting blocks for how to group documents. The Water Process. Think about how the document works in relation to politics, economics, imperialism, nationalism, humanitarianism, religion, society culture, intellectual development advancement. Pretty much every single document the CollegeBoard ever created can fit into one of these buckets. 6. Assess the authors perspective: As you work your way through the documents and group them, keep a few clear questions in mind, Why is the author writing this? What perspective is he or she coming from? What can I tell from his or her background? Asking yourself these questions will help you ensure part of your thesis and essay integrates bias and analysis of bias. 7. Society. Read the historical background: The little blurb at the beginning of the document isnt there for difficult mother daughter relationship, no good reason.

The historical background section of AP European History is like the freebie slot on a bingo cardit will reveal to mockingbird society, you the what issues, time period of the document and allow you to gain a little perspective into the point of view of the source. 8. Connect between documents: The difference between scoring a perfect score on society your essays and scoring an almost perfect score can often come down to your ability to relate documents with one another. As you outline your essay, you should think about at least two opportunities where you can connect one document to another. So how do you connect a document? Well one way would be writing something along the lines of, The fact that X person believes that XYZ is the root of XYZ may be due to the fact that he is ice skating, Y . So in this example, I may pull X person from document 1, but use document 4 to mockingbird, support my Y of the reason why he thinks a certain way. When you connect documents, you demonstrate to what ethical in counseling, the grader that you can clearly understand point of views and how different perspectives arise. It also is a way to demonstrate your analytical abilities. 9. Mockingbird. Start practicing as early as possible : AP European History isnt quite like AP World History where you can get away with just understanding key trends and patterns.

Because the test is much more detailed-oriented, you need to start practicing at least a month and a half prior to your AP European History exam date. Go to AP Centrals homepage for AP European History and select a few essay questions to tackle for the weeks leading up to the exam. Try to tackle two to five a week. Find a proctor like a sibling, parent, or teacher and have them simulate the test for you under timed conditions. 10. What Issues In Counseling. Do not blow off the DBQ : In 130 minutes, 50% of your AP European History grade is determined. Mockingbird Society. In case you didnt know the AP European History exam is a 50-50 split between multiple choice and free-response questions.

Students often overlook the importance of the DBQ and FRQs. Dont be that student. Did you know if you got 0/80 multiple choice questions right but scored 9s on Teaching Children your FRQs and your DBQ, you would still get a 3 based on the 2009 exam curve? Its crazy, but its true. 11. Print out your writing : Writing a coherent essay is a difficult task. In order to do this successfully on mockingbird society the AP European History test you want to make sure that you have spent a few minutes in the very beginning of the test to properly plan out an outline for is division, your essay. You may have heard this advice hundreds of times from teachers but the reason why teachers give it is because it really does help. Ultimately, if you go into your essay without a plan your essay will read without a sense of flow and continuation. One of the things you are assessed on is your ability to create a cohesive argument.

12. Society. Organizing with chronological order : One way that you can order some essays is by difficult daughter, using chronological order. When you frame your argument around chronological order, you want to look for transition points and mockingbird, use those as an opportunity to start a new paragraph. 13. Compare and contrast : Sometimes on the AP European History test youll be asked to compare and contrast. In this case a lot of students simply compare but they do not contrast.

Make sure that you allocate at least one paragraph for each component. 14. Refine your thesis : Crafting the Purification Process Essay, van Gogh of thesis statements can be difficult when under a time crunch. But dont worry the good thing is that if you create a general thesis statement to work off of, you can go back and refine your thesis statement at the very end. Dont be afraid to come up with the general idea and go with that; then at the end of the mockingbird society, paper, revise your original thesis around the main arguments that youve made throughout. Start your AP European History Prep today. AP European History Multiple Choice Review Tips. 1. Read continuously : Heres the thing about AP European Historyits incredibly detailed-oriented. That means its not quite like some other AP tests where you can just cram two nights before and get a 5. In order to really understand connections in European History, you need to what in counseling, keep up with your reading throughout the school year. This not only applies to mockingbird, help you in and Heart: Day and Romero the multiple choice section, but also in the essay portion to understand what time period the mockingbird, prompts are coming from.

Viaults Modern European History should be like your bible when it comes to reading about AP European History. 2. Ultrafiltration: Essay. Identify and hone in on your greatest weaknesses : When you start practicing multiple choice for AP European History, youll quickly realize that there are certain time periods and things you know like the back of your hand, and society, others that are just very hazy to you. After you have had a practice session with AP European History multiple-choice questions , write down the areas where you struggled and review those sections of your class notes. Ethical In Counseling. Make flashcards and review 15-20 every night before you go to bed. 3. Supplement your learning with video lectures : While YouTube can be a distractor at times; it can also be great to learn things on the fly!

Crash Course has some great videos here pertaining to AP European History. Use them to affirm what you know about certain time periods and to mockingbird society, bolster what you already know; then, practice again. 4. Is Division Of Labor. Hanks History Hour : Going along the lines of alternative ways to learn AP European History, you can also learn a great deal from Hanks History Hour, which is a podcast on different topics in history. This is a great way to mockingbird, actually go to sleep since you can listen to the podcast while you dose off. Did you know when you go to sleep you remember what you heard last the best when you wake up? 5. Answer every question : If youre crunched on time and difficult mother relationship, still have several AP European History multiple-choice questions to answer, make a solid attempt at society, answering each and every one of them.

With no guessing penalty, you literally have nothing to lose. 6. Create flashcards along the of labor, way : After you have gotten a multiple choice question wrong, create a flashcard with the society, key term and the definition of that term. Think about potential mnemonics or heuristics you can use to poem, help yourself remember the term more easily. One way is to mockingbird, think about an outrageous image and to Home Romero Essay, associate that image with the term related to AP European History. 7. Use the Process of Elimination : When it comes to tackling AP European History questions, the process of elimination can come in handy if you can eliminate just one answer choice or even two, your odds of getting the question right significantly improve. Remember there is no guessing penalty so you really have nothing to lose.

8. Dont overthink things : When it comes to mockingbird, answering easy questions, typically the shortest response is also the right response. Easy questions typically have easy answers. Ice Skating. Try not to choose strangely worded answer responses for easy questions. Most importantly, dont overthink things. 9. All questions are the same weight : When it comes to the AP European History test, all multiple-choice questions are weighted equally. Mockingbird Society. That means that you want to make sure that you take your time in the very beginning so that you dont get easy questions wrong.

10. Use common sense : Often times with multiple-choice questions, contextual cues are given that signal the time period that the The Water Process, question is testing you on. Look out for these sorts of clues. Understanding and recognizing when a clue is mockingbird society, given is fundamental to Teaching Children from Culturally and Linguistically Diverse, helping you understand what concepts youre being tested on. 11.

Take advantage of chronology : When it comes to answering the mockingbird society, multiple-choice questions, the questions are actually grouped in sets of 4-7 questions each. Relationship. Practice recognizing when youre at the start and mockingbird society, end of a group. This will allow you to mentally think about the different time periods that are being tested while also staying alert throughout the duration of the test. 12. Understand the progression of question difficulty : The AP European History test is outlined so that the easiest questions are presented to you at the very beginning of the test. Home Day And Romero. However, as you navigate through the test youll realize that the questions get harder and harder. Use this to your advantage. Mockingbird. Stay aware of Home, how much time youre spending in mockingbird different sections of the multiple-choice section. While you want to make sure that you allocate enough time at the very end for answer difficult questions, you really want to make sure that you knock the first 60 questions out of the ballpark. 13. Are Some In Counseling. Study themes appropriately : Generally speaking, the AP European History test dedicates 20 to 30% of the multiple-choice section 2 testing cultural or intellectual subject areas.

The remaining 80% are split relatively evenly between economic and political factors, as well as overall social issues. 14. Use your writing utensil : As you work through the society, multiple-choice section of the AP European history test, physically circle and underline certain aspects of answer choices that you know for fact are wrong. Get in this habit so that when you go back to review your answer choices, you can quickly see why you thought that particular answer choice was wrong in the first place. This is a technique that you can use for Process Essay, more than just the AP European History test. 15. Circle EXCEPT : EXCEPT questions can often throw students off so make sure that you get in mockingbird society the habit of physically circling every time you see the People’s and Heart: Day and Romero Essay, word EXCEPT. 16. Go with your gut : You know what Im talking aboutwhen youre at the end of your test and mockingbird, you go back to that one question that nagged you and ice skating poem, you think that you need to change your answer. Mockingbird Society. Dont.

More often than not your gut was right. Theres a reason why you chose that answer so go with your instinct. 17. Use checkmarks : If you feel confident about your answer to a particular multiple-choice question, make a small checkmark next to Culturally and Linguistically, that question number. The reason why you want to do this is mockingbird society, that when you go back to review your answer choices, youll be able to Ultrafiltration: The Water Process Essay, quickly recognize which questions you need to spend more time taking a second look at. Also, making this checkmark gives you momentum moving forward throughout the multiple-choice section. If you feel good about an answer, that little bit of mockingbird, positive reinforcement will help keep you alert as you move through the multiple choice questions. Start your AP European History Prep today. Overall How to Study for AP European History Tips.

1. Do not read your book for straight facts and is division, figures: The way middle schools teach history set up high school students for failure when it comes to tackling challenging history courses. Mockingbird Society. Rather than memorize facts from ice skating poem, your book like youve done since middle school, create a framework and society, general understanding of the core themes from what ethical, your reading. Mockingbird Society. Believe it or not, knowing the type of bread that XYZ leader liked is not important. A lot of history books go excessively in depth in regards to Transcending People’s Home Day and, the nitty gritty. Learn to selectively read the mockingbird, important bits of information and practice summarizing the key points of your reading by outlining 3-5 key takeaways in your notes on your readings.

If you cannot connect the People’s and Heart:, dots, then you will simply craft essays with random name drops and date drops; as a result, your AP score will reflect your inability to create a cohesive argument. 2. Try out the SQ3R method: This is a popular studying technique that can be applied for more than just AP European History. Francis Robinson originally created it in a 1946 book called Effective Study . 3. Society. S (Survey): Preview what you are about to read. Look at the beginning of the chapter and look at the end. Look at the main headings of each subsection of the chapter. Read the discussion questions often found at the end of what ethical issues in counseling, sections. Think about mockingbird society how this section relates to a larger part of history; think about how this may connect to something youve previously learned. 4. Q (Question): Think about questions to keep in mind as you prepare to what is division of labor, read. One way to do this is by re-framing the headers of subsections and to pose them as questions. Mockingbird Society. Ask questions such as, Why is this important?, What does this reveal to me about the overall time period?, etc. 5. Read (R): Now you can begin to read.

After surveying and questioning, you can now read the The Water Essay, chapter keeping the prep work youve done in mockingbird mind. Doing S and what ethical issues, Q beforehand helps keep you engaged and active. Mockingbird Society. Make sure you use your pencil to guide yourself as you read. If you can write in your book, circle and what, underline key things. Active reading helps the content stick with you. 6. Recall (R): At the society, end of each major section, take a minute or two to recall the key things that you just read about. Review the bolded key terms, and Home Day and, answer the mockingbird, main questions you posed to Teaching Who Come and Linguistically, yourself earlier.

Use your own words to describe what you just read. Think about it like you are telling your best friend about what you read about today. Saying things out loud can help you remember things more easily. 7. Review (R): You can either do this with a friend or by yourself. After youve done SQRR, you want to top everything off with review. Look at the notes youve taken along the way and test yourself on the key bits of mockingbird, information from Teaching Children Who Come from, your reading. The key to the SQ3R method is creating a system of processing information and making that information stick. By reviewing several times at random points of the mockingbird society, day, youll help move the information youve learned from your short-term memory to your long-term memory.

8. Connect, connect, connect: In case we havent mentioned it enough, AP European History is all about connecting the dots. Ultrafiltration: The Water. Whether youre just doing your nightly reading or reviewing for your test, its helpful and essential that you recognize how events and people in history are interrelated. Society. History is the study of how people interact with one another. One technique to ice skating, make sure you are connecting the dots is to write key events or terms on flashcards; then at the end of your reading or review session, categorize your flashcards into 5-7 different categories. You may end up doing this by time period, by a significant overarching event, etc. A good way to think about this is you have 5-7 drawers, and a bunch of random things lying around in your room. Each thing represents some event or important person in mockingbird society history and you want to fit all the things into one drawer in order to make your room clean again. If the clean room analogy doesnt work for you, try to think of Children Culturally Backgrounds, a way to get in the categorizing mindset yourself and let us know about it! 9. Society. Create a cheat sheet: While unfortunately you wont be able to difficult, use your cheat sheet on the actual test, you can use a cheat sheet to help simplify your reviewing process as the AP European History test gets closer. Create a cheat sheet that is flexible and society, can be added on what are some ethical issues in counseling tothen as the mockingbird society, year progresses and you do more and more readings, add to your cheat sheet. Before you know it, youll have a handy and hopefully concise reference guide that you can turn to in those last few weeks before the test.

Tips Submitted by AP European History Teachers. 1. Keep referring back to the question : While writing the essay portion, especially the DBQ, remember to Transcending Home and Heart: Romero Essay, keep referring back to the question and make sure that you have not gone off on a tangent. When students drop the mockingbird, ball on an essay it is usually because they do not answer the question. Thanks for the tip from Ms. Difficult Daughter. N at society, South High School in MI. 2. Review your vocab : Complete the vocabulary at the beginning of each section of your preferred AP European History prep book.

If you do not know the meaning of the terminology in Teaching Children from Culturally and Linguistically Diverse a question you will not be able to answer the question correctly. Thanks for the tip from Ms. Mockingbird Society. O at Northville High in MI. 3. What Is Division Of Labor. Do lots of society, point-of-view statements: You dont want to suffer on your DBQ because you only had two acceptable POVs. Do 4 or 5 or 6. And be sure to say how reliable a source is Romero Essay, ABOUT WHAT based on their background, audience or purpose. Thanks for the tip from Steve! 4. Mockingbird Society. Complete readings as they are assigned: Chunking material is the best. way to learn and difficult relationship, then to mockingbird, synthesize material. Look at the primary sources and secondary sources to support textual readings. Think in thematic terms. Thanks for the tip from Ms.

J at Culturally and Linguistically, Trinity High in PA. 5. Society. Supplement your in-class learning with videos: Tom Richey has put together a comprehensive YouTube playlist just for AP European History students. You can check it out at here. Teaching Children Culturally. He also has a great website you can check out here. 6. Society. Provide context in your DBQ: When trying to write a point of view statement for the DBQ you must include three things: First, state who the author really is. Second, what did he actually say. Third, why is said it. Are you a teacher? Do you have an difficult relationship, awesome tip?

Let us know! Hopefully youve learned a ton from reading all 50+ of these AP European History tips. Remember, AP European History is one of the mockingbird, most challenging AP exams to score high on, so its crucial you put in the work to what, get you there. Read actively and society, review constantly throughout the year, so that you do not feel an incredible burden of stress as the AP exam nears. Approach readings using SQ3R, connect the dots between documents, and understand how you are going to be graded by Teaching Who Come and Linguistically Diverse Backgrounds, AP readers.

Youre going to do great! Good luck. Start your AP European History Prep today. Looking for AP European History practice? Educators, are you looking to improve student outcomes? Rigorous, standards-aligned interactive learning for all AP courses. College Entrance. Up-to-date learning supplements for SAT and ACT. Differentiated Foundations. Math, literacy, and writing supplements for school-wide implementations. Mockingbird Society. NGSS Engage.

An innovative CCSS-aligned supplement to accelerate NGSS adoption. Do lots of point-of-view statements: You dont want to suffer on Transcending People’s Home Romero your DBQ because you only society had two acceptable POVs. Daughter. Do 4 or 5 or 6. And be sure to say how reliable a source is ABOUT WHAT based on their background, audience or purpose.` DBQ Nail the thesis! Once the docs are grouped be sure to follow the instructionswrite a clear detailed and precise thesis that addresses all parts of the question. We definitely agree, Keith! Answering the question (#1) and mockingbird, refining your thesis (#14) are crucial. Thanks for this post and its good post. Please continue to sendvery helpful.

Glad you enjoyed! While some aspects of this were very useful, the full capacity of this list could have been better utilized if it wasnt sent out so close to the exam. For example, several of the points stressed keeping up of your reading and material assignments, but for some, its a little to late to tell them that. *EDIT: The tab to show me this list didnt appear towards the end of the year, so if it were made known earlier, it would have been more useful. *AP and Advanced Placement are registered trademarks of the College Board, which was not involved in the production of, and Children Backgrounds, does not endorse, this product. SAT is a registered trademark of the College Board, which was not involved in the production of, and does not endorse this product. ACT is a registered trademark of ACT, Inc., which was not involved in the production of, and society, does not endorse, this product. GMAT is a registered trademark of the Graduate Management Admission Council, which was not involved in the production of, and does not endorse, this product.

GRE is a registered trademark of Educational Testing Services (ETS), which was not involved in the production of, and does not endorse, this product.

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america essay topics When writing an mockingbird society, essay, the first problem you might face is deciding on what topic to are some issues choose, which is pretty ironic if youre writing a problem-solution essay. Mockingbird Society. The way out of that dilemma is to what are some in counseling choose an issue that youre really passionate about. Society. You should also select a problem that has a viable solutionsomething with actionable measures that readers can take. Teaching Who Come And Linguistically Diverse Backgrounds. If youre still stumped for ideas, then take heart. In this blog post, I offer 20 problem-solution essay topics to mockingbird society help you get started. Each topic will include links to sample essays to give you more ideas.

In each example, I present specific problems broken down by People’s Home Day and Romero the issues of today (political, social, environmental, etc.) and mockingbird ask questions that will help you consider different ideas on how they might be solved. Each problem-solution topic I present offers broad possibilities, so youll have to do the important work of hunting down the Culturally Backgrounds facts and examples to provide specific solutions. Ready? Lets get started! Problem-Solution Essay Topics Social Problems. Proposing solutions to social problems might seem challenging, especially when the issues seem so entrenched. Yet, the point of a good problem-solution essay is to mockingbird society suggest solutions that are actionablesomething that your readers can do. Addressing specific problems will lead to difficult mother daughter specific, well-articulated solutions and to the most interesting and compelling essays. Society. Following are five problem-solution essay topics that touch on social problems.

1. Is Division Of Labor. Problem Undocumented Immigration. Solution #1: Should the U.S. Society. government step up its deportation of undocumented immigrants, or should it offer asylum for those currently living in ice skating, the country? Solution #2 : How should the government address trade policies that affect the society economies of South American countries and Essay lead to mockingbird immigration? Solution #3: Will ending the War on Purification Process Essay, Drugs provide some relief to the rise of undocumented immigrants arriving in mockingbird society, the United States? Should there be more coordination between the U.S. and The Water Process Central American countries in mockingbird, how they deal with drug policies, and if so, in what ways? 2. Problem Sexual Assaults on College Campuses. Solution #1: What steps should universities take to prevent sexual assaults on campus?

Solution #2: Should colleges provide safe places for victims to what is division of labor report rapes, and if so, how? What responsibilities should the administration have in protecting victims identities once theyve reported the society crime? Solution #3: In what ways should students be educated about ice skating, rape culture and mockingbird what responsibility should colleges have in providing that education? Solution #4: Should student social services address rape culture? Should fraternities and of labor sororities be more involved in society, educating students on poem, rape culture? In what ways? Solution #1: What role should mental health providers play in determining prevention? What policies or programs should be enacted that will provide greater care for society people suffering from Transcending People’s and Heart: Day and, mental health issues? Solution #2: Should violence in the entertainment and gaming industry be addressed for a possible influence? How about the news media? If so, how?

Solution #3: What kind of gun control laws should be enacted? What role should gun manufacturers have? Should technology be used to decrease mass shootings? If so, in what ways can it be used? Solution #1: What are the possible ways in which communities can force local governments and police departments to address police brutality? Solution #2: What role should voting play? Should communities have a say in mockingbird society, who heads the police department? What other ways should communities be able to review how local law enforcement polices communities? Solution #3: In what ways should the police department address cultural attitudes among officers about the people they police? Solution #4: What role does militarization in police departments play in police brutality? Should the federal government supply local police departments with military weaponry?

If not, should there be a law against ice skating poem, it? How should such a law be shaped? Solution #1: What obstacles prevent people from getting help? Are there ways in mockingbird society, which those obstacles can be addressed? What about People’s Home and Heart: Romero Essay, social pressure, such as shaming? Should the public be educated about mockingbird, suicide, and if so, how? Solution #2: What roles should schools, colleges, police, social welfare, or other institutions play in recognizing those who are at Culturally and Linguistically Diverse Backgrounds risk and how would they go about mockingbird, doing that? Solution #3: Should there be more therapy programs that are accessible for people? What about mental health programs for Transcending Home and Heart: Day and Romero Essay people who cant afford them? In what ways should they be made available as well? Example Problem-Solution Essays Social Issues.

Problem-Solution Essay Topics Economics. Everyone is mockingbird, affected by the economy in Purification Process Essay, one way or another. Either directly through personal debts, indirectly through the loss of mockingbird society, tax revenues that provide services for everyone, or through an difficult, uncertain job market. A problem-solution essay that addresses economic problems are compelling precisely because everyone is eager for mockingbird answersespecially college graduates. Is Division. Following are six problem-solution essay topics that touch on economics. Solution #1 : What policies should the government enact that will help eliminate or lessen the burden for mockingbird students once they graduate? Should there be greater consumer protections for is division students against predatory banking and credit loan institutions?

Solution #2 : Should universities and colleges bear a greater responsibility in protecting students from prohibitive debts? If so, how? Solution #3 : Should state universities and colleges be allowed to enforce or raise student fees? If not, what would be the mockingbird alternatives to what ethical issues in counseling paying for administrative costs? Solution #1 : Should there be programs on the local level that address long-term unemployment? If so, what? What laws or policies should the mockingbird federal government enact that will address long-term unemployment? Solution #2 : Should corporations take the Process lead to mockingbird creating new job markets? If so, how? How should the what are some government encourage corporations to open up new markets? Subsidies?

Tax breaks? Solution #3 : What should individuals who are in long-term unemployment do? What programs should be accessible to them while they seek jobs? What about unemployment benefits? Should benefits be modified to address long-term unemployment as well as short-term?

Solution #1 : Should the government pass more rigorous consumer protection laws that will regulate and prosecute predatory banking institutions or credit loaning companies? Solution#2 : Should the government provide debt relief programs? If so, how should they work and for society whom? Should non-profit, non-governmental organizations, such as Occupy Wall Street, provide relief or should the government create and enact such programs? Solution #3 : What steps should individuals take to get out of debt? What programs, if any, are available for them? If none, what should be available? Solution #1 : What laws or policies can world governments enact that will address child labor? Solution #2 : Should the Transcending People’s United States government enact trade policies that will address the mockingbird society problem? If so, what kind of policies should it enact? Solution #3 : Do U.S. companies exploit child labor, and if so, should local or state governments punish companies that do?

What should citizens do? Boycott? Pressure Congress to pass laws or prosecute? Solution #1 : Should the government pass laws that protect workers? Should the ice skating poem government pass stricter laws that protect unionization? Solution #2 : How should unions go about mockingbird, encouraging more people to join unions? What about workers who are unable to organize in their workplace?

What steps should they take to mother organize? Solution #1 : What policies should be enacted that will ease economic problems leading to home foreclosures? Should the federal government enact laws that will protect homeowners, and if so, how? Solution #2 : Should local governments pass laws to protect homeowners against foreclosures? What alternatives are available for local governments to mockingbird society prevent them? What should they do with homes that are underwater? How should they prevent blight? Solution #3 : Should non-profit groups like Occupy Wall Street help people fight against ice skating poem, foreclosures, and if so, how? Problem-Solution Essay Topics Politics.

Perhaps this is largely because people get distracted by how challenging the problems are before they can even think about solving them. The key to mockingbird writing a good problem-solution essay is to The Water Purification think small. In other words, pick a very specific problem (money in politics, for instance) that will lead to goals that are clear and viable. When you pick a topic that readers feel confident that theyll be able to tackle, youll write an essay that just might move them to mockingbird society act. Following are four problem-solution essay topics that touch on Home and Heart: Day and, politics. Solution #1 : What leads to mockingbird the dependence on money in electoral politics, and is division what can be done to address the problem? For instance, what alternatives are available for candidates to mockingbird society raise funds for elections?

Solution #2 : Should the government regulate how much money is spent in campaigns? If not, what alternatives are available that will lead to campaign finance reform? Solution #3 : Should the government pass laws that will define who should or shouldnt be able to donate campaign dollars? If so, how should such a law be shaped? How should free speech rights be taken into consideration?

Solution #4 : Should the difficult mother relationship constitution be amended to mockingbird society address the problem? If so, what should be amended and how? Solution #1 : Should there be more vigorous laws that protect Americans privacy rights from government surveillance? If so, how? Solution #2 : Should Congress be more proactive in monitoring the intelligence community? If so, how should Americans be certain that they are? In what ways should citizens be involved in poem, the process? Solution #3 : Do Americans have the right to know what the intelligence community is doing? If so, in what ways can that be done while protecting national security?

Solution #1 : Will electoral reform address the society problems caused by partisanship? If so, in what ways? Solution #2 : In what ways do American citizens help create partisanship? The media? How should Americans be better educated about their roles as citizens? Solution #3 : Will media reform help address the problems that cause partisanship, and if so, how? Solution #4 : How should political parties address partisanship?

Should third parties be allowed to have their voices heard in the electoral process? If so, how? 15. Problem Voter Disenfranchisement. Solution #1 : Should the federal government pass laws that will protect voter rights? Should the constitution be amended to protect voting rights for all citizens? Solution #2 : How should state governments prevent partisanship from Ultrafiltration: The Water, affecting electoral board policies? What should the mockingbird society public do to is division of labor fight against Voter ID laws or other laws that disenfranchise voters? Solution #3 : What steps should be taken to revive peoples faith in mockingbird, the political process?

Who should enact these steps? The public? Schools? The media? Politicians? If so, how?

Solution #4 : Should congressional rules, policies or social culture be changed to discourage and Day and prevent obstructionism? If so, in what ways? Problem-Solution Topics Environment. By all measures, the environment is our most precious resource, yet we face many problems in mockingbird, trying to daughter protect and preserve it. A problem-solution essay that addresses environmental problems can be compelling and thought-provoking because it will alert readers to the necessity of proposing real solutions that people can enact as individuals or as political groups. Mockingbird Society. Following are five problem-solution essay topics that touch on the environment. Solution #1 : What kinds of The Water Process Essay, laws or policies should the government pass that will address climate change? Solution #2 : Should the government push for society more trade policies that will address climate change, and if so, what kind?

Solution #3 : Should foreign policy play a role in addressing climate change? For instance, should the United States work with other major-polluting countries like China, and if so, how? Solution #4 : How should the marketplace address the problem? For instance, should corporations pursue fuel alternatives like green technology? If so, how should they be encouraged to do so? Solution #5 : How should grassroots organizers push for a change in policies? Who would be targeted for Transcending People’s and Heart: Romero Essay such a movement?

The U.S. Mockingbird Society. government? The U.N.? Corporations? Solution #1 : Should the Transcending People’s Home Day and Essay government pass laws that make fracking illegal? Should it promote energy fuel alternatives, such as green technology? If so, how? Solution #2 : How should companies that use fracking be discouraged from mockingbird, doing so? Should they be subject to civil lawsuits? What about Ultrafiltration:, boycotts, civil disobedience, or other grassroots organizing?

Solution #3 : What should be done to educate the mockingbird society public about fracking? Should the news media report on it more often? If so, how should environmental groups push the media to do so? Solution #1 : Should the government pass laws or policies that provide greater protections for preserving and protecting wildlife? If so, what types of laws? Should it go after corporations that endanger wildlife. If so, how? Solution #2 : Should corporations take a lead in The Water Purification Process, protecting wildlife? If so, how? Solution #3 : How should environmental groups address endangered wildlife? What are some of the things they can do to push the government and mockingbird corporations to protect the poem environment?

If so, how? 19. Problem Environmental Pollution. Solution #1 : What should the government do about society, pollution? How should it be involved in long-term protections? For instance, should the The Water Purification Essay government set aside relief funds or economic restorations for affected areas? Solution #2 : What role should local and state governments play in protecting wildlife from pollution? Should local governments be stricter in regards to environmental studies for local projects, such as the building of mockingbird, chemical plants or factories near wildlife or residential areas? If so, how? Solution #3 : Should governments pass stricter laws that prosecute corporations that pollute? If so, how should the public push for such laws to get passed?

20. Problem Environmental Injustice. Solution #1 : Should local governments do more to protect communities from environmental injustices? If so, how? If not, what can the public do to fight against them? Will grassroots organizing help? If so, how?

Solution #2 : Should the Home Day and Romero Essay federal government provide relief for communities affected by mockingbird environmental injustices? If so, in Transcending Home and Heart: Day and Essay, what way? Should the Justice Department get more involved in prosecuting corporations or local governments responsible for these injustices? Solution #3 : Should the mockingbird society media report more on environmental injustices? What can the public do to push the media to Transcending People’s Home Day and Essay cover these stories?

How should grassroots organizations get the information out to society the public? Documentary films? YouTube? Crowdsourcing? Example Problem-Solution Essays Environment. Hopefully these problem-solution essay topics will make it easier to get started on your paper. If you need any more help, I recommend reading the blog post One Persuasive Writing Technique You Cant Write Without to get that extra boost to your writing style.

Also dont miss Problem-Solution Essay Tips from relationship, a Kibin Editor. Mockingbird Society. Good luck and happy essay writing! Psst. 98% of Kibin users report better grades! Get inspiration from over what is division of labor 500,000 example essays.

About the Author. Cynthia C. is a short story writer, blogger, former copy editor, and mockingbird newly-minted Doctor Who fan. You can find her on what is division of labor, Google+

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English and Comparative Literary Studies. This handbook is a guide that I’m hoping will enable you. It is geared, in particular, towards the seventeenth-century literature and culture module but I hope you will find it useful at mockingbird other times too. I would like to stress, though, that it is not the only way to do things. It may be that you have much better ideas about what makes for a successful essay and have tried and tested methods of executing your research. There isn’t necessarily a right way and difficult, so I hope you will not see this as proscriptive and mockingbird, limiting. You should talk to all your tutors about what makes for a good essay to get a sense of the different ways that you might construct an essay.

1. Essay writing (p.2) 2. Close reading (p. 4) 4. Constructing an argument (p. 8) 5. Help with this particular assessment (p. 9) 6. Grade descriptions (p. 10)

1. ESSAY WRITING (and historicist writing in particular) Essay writing has four stages: reading, planning, writing and proof-reading. Excepting the last, you may not find that they are not particularly discrete but rather interlinked and mutually informative. If any stage is skipped or done badly, though, it will impair your work. 1) Read the text and make sure you understand it. People’s And Heart: Romero Essay! Use the Oxford English Dictionary online to look up any words you don’t understand or if they are operating in society, an unfamiliar context.

Available on the Warwick web: http://www.oed.com. 2) Do a close reading. Make a list technical features (cf. the page in this booklet entitled ‘close reading’; refer to the section on poetic form in the back of your Norton Anthologies pp. 2944-52). Ask yourself: ‘how does the of labor, text achieve its effects?’ Then ask yourself: ‘how do those poetic effects relate to the meaning of the mockingbird society, text?’. 3) Do some research, particularly on the historical theme, period, cultural group that you’re interested in. You could begin with a general history and then do a literature search for more specialist books and Teaching Who Come Culturally and Linguistically, articles. Society! It may help you to narrow your research to a particular theme or idea that is suggested, hopefully by mother daughter relationship, your reading in mockingbird society, 1) and 2). Rather than trying to find out about the whole of seventeenth-century culture, limit your research to the restoration, cavalier culture, medicine, the family or whatever. (See the handout on Children Who Come Culturally and Linguistically Backgrounds research). 4) Be careful when you take notes so that you will make no mistake, when you come to writing and referencing your work, about what is your work and what is someone else’s. Mockingbird! Read and be clear about the university’s rules on plagiarism which are laid out in Transcending Home Romero Essay, the blue booklet ‘Essay Writing and mockingbird, Scholarly Practice’ which you can get from the general office.

1) Begin by making a spider plan of all your ideas and the relationships between them. IF YOU DON'T LIKE SPIDERS FORGET THIS BIT. 2) Then write out a paragraph (which you will not include in difficult mother, your essay necessarily) called ‘MY LINE OF ARGUMENT’. Society! This will be information to yourself (so it can be very boringly and functionally written) about what you intend to People’s Romero Essay say. Ideally this should be a single big idea, which you can sustain for the length of the essay, made up of stages that can be demonstrated with reference to mockingbird society the passage in question. It may well be that you want to write something similar to are some issues in counseling this ‘line of argument’ paragraph, only in mockingbird, a more dynamic and elegant way, for are some ethical issues in counseling, your introduction. See the page entitled ‘constructing an society, argument’ that has an example of a ‘line of argument’ paragraph. 3) Then write out is division, a linear plan of your essay with a logical ARGUMENT, an argument that is assertively stated and then proved through the course of your piece. TIP: try not to separate out style, content and context; discuss them together to show how the relate to one another.

You are aiming to produce something that identifies and describes both the wood and the trees; indeed, the trees are your evidence for the existence of the wood! You need to put together a big argument out of lots of mockingbird, bits of evidence. 1) Everyone has his or her own way of writing. I sometimes find it easier to write the difficult, middle of the essay first and society, then come to the introduction last, which is perhaps the hardest bit to Teaching from Culturally and Linguistically write. You may find that your ideas change and are worked out mockingbird society, more fully as you start to write. In which case go back to Children Who Come from and Linguistically Diverse Backgrounds B) and produce another plan. Present your ideas as a finished thought, rather than a thought process. 2) Keep yourself closely to society your argument by imagining your reader. Perhaps a friend, a tutor or a parent might serve: imagine them behind you as you write asking ‘SO WHAT?’, making you insist on its relevance and what, trying to prove a particular point. Imagine that you are a newspaper editor writing a polemic, trying to convince your readership of a particular point of view. 3) Inventing a title and society, writing an introduction.

You should try to make your essay interesting to an examiner. Which do you think is the best of People’s Essay, these three titles: ‘Aphra Behn’s Oroonoko’; ‘Discuss the question of race in Aphra Behn’s Oroonoko’; ‘The “gallant slave”: the idea of the noble savage in Aphra Behn’s Oroonoko’. Similarly with the introduction. Society! The first sentence should grab the examiner immediately. Which is a better first sentence: ‘Aphra Behn’s Oroonoko was published in 1688 and is a prose work about Surinam’; ‘At the heart of Aphra Behn’s Oroonoko stands the deep paradox of the ‘royal slave’. 4) Using secondary literary criticism. It is, of course, good to read lots and to incorporate that reading into difficult mother daughter your work. What you are attempting to mockingbird society do, though, is to position your independently arrived at ideas in relation to other critics in the field. You shouldn’t be deferential or let the ideas of Culturally and Linguistically Diverse, others drag you off course.

You should USE other people’s work in the service of your own argument. Mockingbird Society! For example, you might disagree with a critic; you might apply their theory about one text to another; you might say that their work hasn’t gone far enough in its assessment. Never use a quotation from someone else to clinch an argument: just because someone famous has said x or y it doesn’t mean that it’s necessarily true. I sometimes find it useful to write a draft of my paper that includes no secondary reading at all, basing it just on my general knowledge of the critical field. I then do some detailed research in ethical, secondary criticism before writing a second draft. This means that the agenda is not dictated by other scholars, and ensures that I use them rather than becoming their spokeswoman. Make sure, of mockingbird society, course, that all your reading is properly referenced to avoid a charge of Ultrafiltration: The Water Purification, plagiarism. 1) Check the spelling: in particular the names of the author and the text that you’re looking at mockingbird society MUST be spelled correctly. 2) Check your punctuation. If you don’t know how to use particular punctuation marks please get a book and learn how.

In particular the poem, misuse of mockingbird, apostrophes is poem deeply irritating to an examiner. The Collins gem guides are really good also Lynne Truss, Eats, Shoots and mockingbird society, Leaves is fun and Transcending People’s Day and Romero, informative. 3) Make sure that you get hold of the blue booklet, ‘Essay Writing and Scholarly Practice’, from the general office. You must use the reference guide in there. I favor the MHRA guidelines; you may prefer the MLA style. If you do reference a website it is best to put it in a footnote rather than the text were it looks ugly.

You should always include some close detailed analysis of the literary text(s) that you’re discussing in your essay. Society! This demonstrates your sensitivity to the forms, textures and ideological purpose of Transcending People’s Home and Heart: Romero Essay, language. You should aim to show the mockingbird society, relationship between form and meaning, between the text and its world. Before you can put together an argument about the ethical, relationship between a text and its time you will need to do some close reading, compiling a list of technical features in a text or an society, excerpt from a text. Is Division! Choose excerpts that relate to themes or passages that interest you. Then you can develop a checklist of mockingbird society, features to look for. Use this as a guide but you may want to of labor add to, or amend it. *** What you see will be very different from mockingbird, what other people see. So, although it looks like a slightly dry exercise, this is where your ideas, your originality will come from. Close reading, in any module, will make your essays sparkle. *** #61623; Prose, drama or poetry?

#61623; Genre? (e.g. is it panegyric, epic, restoration comedy or what ever) #61623; Does it remind you of anything? Can you compare or contrast it with something of a similar date? Or, alternatively, compare it with something of a similar genre from the what ethical in counseling, previous or next decade, for example, in order to investigate change over time. #61623; Poetry: metre, rhythm and mockingbird society, rhyme. Look at Home Day and the section on poetic form at the back of the Norton Anthology (p. 2944) and other guides. Don’t just describe metre etc but ask yourself how it works in that particular passage.

How are units of mockingbird, meaning created by the line divisions? When a poet downplays or emphasizes a particular word through positioning it in a particular way, what effect does it have? How does the daughter relationship, poet manage tone, pace and register with his use of rhyme and rhythm? iF THESE FEATURES ARE NOT IMPORTANT IN YOUR PIECE IGNORE THEM. #61623; Drama: look at the length / speed of the speeches, the stage directions, the entrances and exits. #61623; Prose: rhetorical features and clause structure are the things to look out for in particular. Are the sentences complex or simple? Is it in mockingbird society, hypotaxis or parataxis?

What about word order and syntax, is there anything unusual or unexpected there? #61623; What is the overall structure of the passage / text? Are there abrupt changes or a progression from one idea to is division of labor another? #61623; What other structures are there? Symmetries, comparisons and mockingbird, contrasts, digressions, asides, repetition. Transcending People’s Home Day And Romero Essay! Is there any dialogue? Are the arguments circular or progressive? #61623; Are there any words you don’t fully understand? If you aren’t in a closed exam you could look them up in the Oxford English Dictionary online. This would also give you a sense of the mockingbird society, other meanings that that word might have.

Are there any puns? #61623; Think about grammatical features: tenses, conditional constructions, the ice skating poem, passive voice. Is the passage in the first, second or third person? Perhaps there are tense or person shifts; what effect do these produce? #61623; Look out for society, predominance: several superlatives or comparative adjectives and Teaching, adverbs; a lot of words that mean a similar thing, repetitions of possessive pronouns or what ever. #61623; What kind of language is being used? i.e. what register is it in? Is it elevated or earthy, legal or lyrical, rhetorical or religious?

Why? #61623; Look for particular rhetorical features: metaphor and simile, hyperbole and society, litotes, personification, metonymy and so on. #61623; Look at punctuation (but be careful: it could be the intervention of ice skating, a printer or a later editor). Look out for: enjambment, parentheses, direct speech? When the society, punctuation is sparse, why? Is it because there is a proliferation of conjunctions that resist punctuation like, for example, the word ‘and’. This may indicate parataxis or a very conversational style. #61623; Look out for allusions and what ethical issues, references, often to the bible or classical stories. If you don’t know them and you’re not in a closed exam, look them up in a reference dictionary or on the internet. #61623; What is the tone of the passage? Is it homiletic, comic, anxious, melancholy or ironic?

How is this effect created? #61623; Where else does that poet use similar phrases, ideas, patterns and images? What does it say about society, his or her concerns and art? TIP: Don’t make simple associations between sense and sound. For example, whilst there are a lot of warm words that begin with ‘m’ (like, for example, milkmaid, mother, magic etc) there are also some, like ‘malice’, ‘muscular’, ‘murder’ which evoke quite different associations.

You then need to think how those technical features, which you’ve noted construct the meaning of the passage / text. Do not think about form and content as separate things as if form were a kind of cloak in Ultrafiltration: Process Essay, which meaning is dressed: they are organically connected. Above and beyond that you will also need to think about how that text (both its form and its meaning) relate to the particular concerns and fashions (literary, political, philosophical etc) of its time. Mockingbird! You might think about the way in which repeated ideas in your text / excerpt link to significant contemporary discourses. Look for substituted vocabularies: i.e. when love / sex is discussed with the language of money / credit for example. Could that be related to prevailing economic trends and ideas? When you are constructing your ARGUMENT and writing your essay, consult your close reading list. Not everything there will be relevant to your ARGUMENT; you only want to Teaching from Culturally and Linguistically Diverse Backgrounds include the mockingbird society, things that relate, that offer evidence for a particular point of view about what are some ethical issues, how the text is placed culturally, politically, socially and / or historically.

Research is crucial for any essay and mockingbird society, requires a certain amount of initiative. You will partly have to learn by trial and Teaching Children from Culturally and Linguistically Backgrounds, error. Here are a few tips and ideas, though. Read both narrowly (and address the theme of mockingbird, your essay) and Diverse Backgrounds, also widely. So if you are, for example, researching infanticide, also research the family or law / crime. When you research a context it might be worth look at the work of society, philosophers, painters, and theologians and see what they were saying / doing in this period. An essay which looked at the early modern patriarchal family in the light of People’s Home Day and Romero, Robert Filmer’s political tract Patriarchia, for example, would be much more interesting than one that only looked at modern historians’ account of the mockingbird society, early modern family. Mother Daughter! An essay that discussed the mockingbird, panegyric written to, or on a particular king, alongside the portraits that were painted of him could also be very suggestive. EEBO might be very useful here at helping you to mother find out about, say, sermon culture or advice literature. (look at the last page of society, this booklet for some help here). Think of some the areas, themes, historical moments, authors and ideas that you want to find out Transcending and Heart: Romero, about. List them as key words.

For example: Aphra Behn, Oroonoko, race, royalism, restoration, early modern, colonialism, slavery etc Do not be limited here. Think of terms / phrases that will give you some background too. How about society, ‘cheap print’, ‘renaissance politics’ etc Then begin on the computer. Be careful of what is division of labor, stuff that you find on the ordinary WWW. It is not usually very reliable. Often this is stuff that people can’t publish in proper books.

Use it is a guide and be very critical. 1) http://www.jstor.org (through the Warwick network only). Here you can read articles from reputable, peer-reviewed journals on society line. Transcending People’s Romero Essay! An excellent starting point. Try various combinations of your search terms in mockingbird, either the Basic search (will give you hundreds of what, items) or in the advanced search form (which will give you much narrower and mockingbird society, probably more useful stuff. Try it out; go to the advanced search form: A) In the box marked ‘All of these words’ insert the word ‘Behn’. Then tick the box marked ‘title’ and then also the box marked ‘article’. Press the ‘Search’ button. See if you can identify any articles with a particularly historicist bent. B) In the ethical issues, box marked ‘All of these words’ insert the words ‘White’ and ‘Black’ and ‘England’.

In the box marked ‘exact phrase’ enter ‘Seventeenth-century’. Press search and see what you get out. Try other, similar search terms. C) In the box marked ‘All of these words’ insert the society, word ‘Royalist’. In the box marked ‘at least one of these words’ enter the words ‘print culture’.

Perhaps limit to articles by checking the relevant tick box. Press search and see if any of those are useful. [you will see that sometimes you have to do some considerable sifting to find good things.] 2) The Modern Language Association of America database direct access from the Warwick network at: http://www2.warwick.ac.uk/services/library/electronicresources/databases/#m. The bibliographic databases are listed alphabetically so scroll down to ‘M’. Select ‘MLA’. This will give you the Transcending Home Romero Essay, reference only (although Warwick may provide a link to the on-line journal). You may find that some of the mockingbird, things that are listed you won’t be able to get because Warwick doesn’t subscribe to what that journal or perhaps the item is a doctoral dissertation from society, another institution. Don’t worry, you’re not expected to read everything under the sun. Difficult Daughter! Leave those things that you can’t get. Try it out: put in the search terms ‘Aphra’, ‘Behn’ and ‘race’ into society the keywords box. Press search and see what you get.

3) Historical abstracts: http://www2.warwick.ac.uk/services/library/subjects/arts/elecresources/#databases_internet. Again, use this database to help you compile a list of articles or books that you could look at either on-line, if Warwick has a link, or in the library. Ignore the Teaching Who Come Culturally and Linguistically Diverse, things that you can’t get hold of. A) Put the search terms ‘restoration’, ‘race’ and ‘England’ into the keywords box. Press search. B) Put the search terms ‘early modern’ and ‘print culture’ into the keywords box. Press search. Again you will have to decide what’s useful / relevant. 4) Use the library catalogue, don’t limit yourself to books about mockingbird, English. Put in search terms that will give you books on the historical background that you’re looking for.

Once you have found one book on the shelf look around in that same area for others that will be related by subject. 5) Look on is division your reading list for general background books. CONSTRUCTING AN ARGUMENT. Producing a successful argument is a process that has a number of stages. Often you will understand your argument better after you have started writing. It is important that you go back and society, re-plan your work, taking into account your new findings. You will need to develop a provisional thesis, however, so that you have somewhere to start: a focus for your close reading and research. You don’t need to argue that history is important for the study of literature. You can take that as a given and move on to say something a bit more sophisticated about how the particular poem / play or prose piece you’re working on intersects with a particular set of of labor, events or ideas in a specific historical moment. A good argument should be fairly specific rather than general and comprehensive.

In particular, when writing a historicist essay, do not list the ways in which one text is embedded in its period. Instead choose one of those ways and research it in mockingbird society, more depth. So, rather than writing about, say, Ben Jonson’s interest in Anabaptists, Spanishness, alchemy, the The Water Process Essay, plague, etc in The Alchemist, choose one of these themes and find out about it in the historiography of the seventeenth century and couple this research with a close reading of those sections of the play that treat that theme. Your readings of the text and the history of the mockingbird society, times should suggest your detailed argument. What Is Division! Don’t think of your argument first and then try to press it onto mockingbird society, the play or poem you’re interested in; allow your idea to mother daughter grow out of your reading. Below is my best attempt at a LINE OF ARGUMENT for mockingbird society, an essay on Rochester and Milton.

Again, I should stress that this is Transcending Home Day and Essay only by way of demonstration what I would do. This is very different from what you would do. There is mockingbird no one way and your ideas will be as interesting / valid as mine. Don’t think that you have to produce something the same, or even necessarily similar I have done this just to ice skating give you an example of what I mean. I have tried to construct an argument which uses both close reading and historical context. Imagining the future in the restoration: a critical comparison of the mockingbird society, poetry of Teaching Who Come Backgrounds, John Wilmot, Earl of Rochester and John Milton.

Line of Argument: This essay will argue that Rochester’s poetry is not only everywhere fascinated by time, regularly exploring what it is and how it operates, but that this interest betrays his sophisticated engagement with contemporary political philosophy. It will closely interrogate the forms of several of Rochester’s time-related poems for their political sensibilities. It will then contrast those poetic forms and political sensibilities with those in the poetry of John Milton and especially Paradise Regained. Milton as I shall show with the use of historical evidence is very differently socially and politically placed, indeed at the other end of the ideological spectrum from the Earl of Rochester. I shall show that the difference is one of dispossession (Rochester) and providence (Milton). Rochester’s narrators exist in fear of, and subject to an arbitrary and absolute future; Milton’s Paradise Regained, on the other hand, asks an imagined republican reader to wait in anticipation of a future in society, which God will deliver their political success. I shall explore the way in from Diverse Backgrounds, which Rochester’s pessimism the idea and tone of dispossession in his poetry and Milton’s optimism the visionary quality of his providential allegory stand in contrast to society the respective fortunes of the political groups to which those poets actually belonged and at Purification Process the particular times when the mockingbird, poems I’m discussing here were written and published: i.e. Rochester’s being part of the royal court and Milton’s being displaced from his office at the restoration of Charles II.

This will arrive at, by is division of labor, way of conclusion, the demonstrable sadness of some of Rochester’s verse which indicates the complex circumspection with which he viewed his own aristocratic, political community and its limited expectations of monarchical authority. HELP FOR THIS PARTICULAR ASSESSMENT. Details of what you are expected to do are on the departmental website at: http://www2.warwick.ac.uk/fac/arts/english/undergrad/modules/second/en228/assessedessay2/ There you will find a list of mockingbird society, texts and details of how to find them on EEBO (Early English Books Online). Their website is at: http://eebo.chadwyck.com/home. You need to download those texts, read them and then choose one to write about. You could also read the essays, published on the EEBO website, by previous Warwick students that have won prizes for Purification Process, their attempts at this assignment. http://www.lib.umich.edu/tcp/eebo/edu/edu_win_03.html. You might also use EEBO in your essay research. Try the mockingbird society, subject list in particular. If you get yourself to the search form at http://eebo.chadwyck.com/search you can click on the link marked ‘select from a list’ next to the subject keyword box.

This has all sorts of interesting categories: look up, for example, ‘anti-catholicism’ or ‘restoration’, ‘credit’ or ‘murder’. I would like you to do what you can in terms of placing the text of your choice, and researching it. Then I’d like you to come and see me at the end of term with a title and a line of ice skating poem, argument. You could also, if you wish, bring a longer essay plan. This is Isabel talking to her group. We will all be available on email over the holidays--do ask. Mockingbird! Gabriel won't be here after the holidays--he lives in Ultrafiltration: The Water, London--but do come and see me, his group, if you need a person to society talk to. USING THE OXFORD ENGLISH DICTIONARY.

The OED can be found online (through the Warwick network) at http://www.oed.com. When we read an edited text we often have a helpful gloss which an editor has provided so that words and phrases that we don’t understand are defined for us. Mother Daughter Relationship! In this assignment you will have to put together that gloss for yourself and the best way to start to do that is with the mockingbird, OED. The OED is an extraordinary resource that will give you assistance in is division of labor, all sorts of ways. For example: a) it will obviously help you to understand words which you don’t understand or unusual applications. It will also help you to find obsolete and dialect words. b) it will help you to mockingbird society see how words have changed their meanings or emphases over time. c) it will help you to Transcending People’s and Heart: Day and Essay identify puns. There may be sexual or religious connotations to a particular word that we may have lost. Some times our modern definitions will co-exist with old, and society, now obsolete meanings.

d) it will tell you the and Heart: Romero Essay, earliest use of a particular word. This is mockingbird society useful for ice skating poem, working out mockingbird society, which of several definitions might apply to the word you’re looking at. Look at the examples, that is the quotations that are given, and note their dates. It may be that you find that the what of labor, word was new or recently borrowed from mockingbird society, another language. Click the Children Culturally Diverse Backgrounds, ‘date chart’ button to see the mockingbird, uses represented on a time line. It may be that you will find that a word is used differently and in different contexts at different points of the seventeenth century: what might the use of a particular word / phrase tell us about an author’s engagement with political, historical or sociological movements? e) Look at Transcending People’s Home and Heart: Romero Essay the etymology: this might tell you about how the text you’re looking at mockingbird engages with particular fashions or imperial encounters. Look up, for example, ‘chocolate’ where does the word come from?

At what period does it come into what ethical issues in counseling the language? f) the examples given in the dictionary will also help you to mockingbird see how other contemporaries used the Teaching and Linguistically, word or phrase you’re interested in, and in what sort of contexts it came up. Mockingbird! In this way it can operate as a concordance. You should investigate the poem, concordances available in the library, by the way. Similarly they will give you a sense of how a particular word or phrase is used elsewhere. You should use the OED not just to look up words that you don’t understand but also other words, especially those that are used in society, an unfamiliar way. You will find more interesting things if you look up lexical, rather than grammatical words. That means verbs, adjectives, adverbs and nouns rather than prepositions, articles and pronouns. You need to remember that there was no standard spelling in ice skating poem, the early modern period; the move to standardize spelling did not occur until the society, middle of the eighteenth century.

This means that when you have a word you don’t understand it you may not get an adequate definition by putting it in exactly as it is into ethical issues in counseling the OED search box. Try that first but if it isn’t found, or you get a definition that is not right (i.e. the examples indicate that its earliest use was a lot later than your text) you should try different spellings. Society! In particular the vowels are often interchangeable. Try every vowel combination that you can think of. Try substituting ts and cs, us and vs and other related consonants. Try out the OED. Look up the following words: how have their meanings have changed? Where do the words come from? How were the words used at difficult mother daughter different points in history? And in the seventeenth century in particular?

Department of English and Comparative Literary Studies, Humanities Building, University of Warwick, Coventry CV4 7AL.