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How to networks, Write Better Essays: 6 Practical Tips. The best students enjoy writing essays because they’re a chance to shine; they’re an interesting intellectual exercise in cover no experience which the writer must craft thoughtful arguments on complex topics within the confines of a prescribed and often restrictive word count. For many such students, each essay brings with it the challenge of making it that little bit better than the last one. Phd Thesis! The problem is that when you write essays regularly, it’s easy to get stuck in a rut of in computer repeating the same formula each time particularly when you already receive good feedback from the teachers who read them. So how do you take your essays to the next level and go from great to brilliant? Here are some practical tips and phd thesis networks techniques that will help you write consistently impressive essays. Offer to phd thesis networks, share your essays with other people and they may return the favour. Phd Thesis In Computer Networks! Even better: start a study group.
Just as the books you read subconsciously help mould your own writing style, so reading other people’s essays can help you develop and build on your own essay-writing style. Try to read a range of other essays, including those of your peers and of academics. Read essays on a wide variety of subjects, not necessarily just those that you’re studying; different disciplines might apply different kinds of arguments or styles, so the wider you read, the more possible techniques there are for you to pick up and use in essays of your own. As you read other people’s essays, don’t just take them at face value. Be critical: what do you like about essay on the self them? What don’t you like about them? How persuasive do you think they are? Is the phd thesis in computer, argument a balanced one, with points adequately supported with evidence? Has the une introduction dans, writer used any techniques you’ve not seen before? Another good source of in computer networks essays is the broadsheet newspapers.
Read the opinion pieces and dissect how the writer has supported their points with evidence, and again, be critical; note where they#8217;ve left things out to try to persuade you to a particular opinion. Essays should be balanced, so you can learn from the best of these writers and pick up some techniques to help you shape a balanced piece. 2. Essays About Responsibility! Build your vocabulary and use it properly. Make use of dictionaries and thesauri. A good vocabulary will allow you to phd thesis in computer networks, express exactly what you mean, as clearly and concisely as possible. Economy with words is a characteristic of all good essays, because readers (and essay-markers) don’t like having their time wasted with long, rambling points that could have been expressed in half the number of words.
One way of statement at the p.o ensuring that you can communicate clearly and to in computer, the point is through accurate and effective use of essay on the who am i advanced vocabulary. A good essay writer should never rest on their laurels when it comes to vocabulary; it’s something you should be working on continually, as there are always new words to learn that could help convey a point more effectively. What’s more, deploying a good vocabulary displays intelligence and allows you to phd thesis in computer, be more persuasive in your essay-writing. Here are some ways in which you can build your vocabulary: Subscribe to a #8216;word a day#8217; email (such as this one from Merriam-Webster). Create a folder in your email account for new word emails, so that you can file each email away and have them all in one place ready to une introduction dans une dissertation, flick through and learn from in an idle moment.
Read widely, and refer to a dictionary for words you don#8217;t know as you go along; this way, you’ll learn the new word as well as seeing it in context so you know how to use it properly. Read different genres of fiction, and phd thesis in computer networks non-fiction covering a range of topics, and you’ll have the added bonus of widening your general knowledge as well as your vocabulary. Use a thesaurus if you find yourself using the same words over une introduction une dissertation and over again, add variety to phd thesis, your language by looking up those words in letter for waitress no experience a thesaurus and phd thesis networks finding other words that mean the same thing. A word of warning: words you find in a thesaurus can’t always be used interchangeably; even words with similar meanings can differ subtly in comment dans une dissertation a way that makes them inappropriate in phd thesis certain contexts, so find examples of cover letter no experience a word used correctly before you use a new word for the first time. Learn prefixes, suffixes and networks roots it sounds boring, but this shortcut will help you learn a great many more words. Many roots come from Latin and Greek words, such as “bene” in Latin, meaning “good”, which gives rise to words such as “benefactor”, “benevolent” and “benefit”. Responsibility! It#8217;s often possible to deduce the phd thesis in computer, meaning of essay self who am i a new word if you know its root and read it in networks context. Prefixes are added to the beginning of thesis for why i live at the a word to phd thesis in computer networks, change the meaning, such as “semi” or “ante”, while suffixes are added to at the, the end, such as “-able” or “-ance”. Start a vocabulary book you probably have one if you’re learning a foreign language, so why not have one for your native language as well? Buy yourself a nice notepad and use it to collect new words and in computer their meanings.
The act of writing down the definition will help you remember it, and you could include an example of how the word is used to increase your chances of memorising it for use in essays. It may help to have different sections for words on particular themes; you could have a general section, and then further parts of the notebook could be dedicated to words of use in history essays, science essays and so on. The aim of improving your vocabulary is to increase precision and reduce waffle. Put the new words you’ve learned to about, good use right away, perhaps setting yourself the challenge of including a minimum number of new ones in networks each essay you write. This will help consolidate your knowledge at the same time as impressing the reader. One important thing to remember, though: don’t use big words just for the sake of it. Using a long, obscure word when a simpler one would suffice risks making you sound pompous, which may have the opposite effect to the one intended. What’s more, be wary of adding words for the sake of it; cut the waffle by reviewing each sentence and removing any words or sentences that don’t add anything to what you’re saying.
Ultimately, your goal should be to with no experience, make your writing as clear and easy-to-understand as possible, so that it is a pleasure to in computer networks, read. 3. Words to phd thesis, help develop an argument. Part of sounding intelligent in an essay is not repeating yourself; as you’re writing, focus on using language effectively to help build an argument and phd thesis in computer create a sense of structure. To that end, avoid using the same words every time; many people overuse the phd thesis in computer networks, word “also”, for example. Vary your language, and use words such as “moreover”, “furthermore” and in computer “however”. Such words help develop your argument and make the networks, reader feel they are being guided through the phd thesis in computer networks, problems on a sort of dual essays ‘journey’ to your conclusion. Would you be able to summarise your essay between floors? We’ve probably all had it hammered into us that we should write an essay plan before we start writing, but before you even do that, you need to know what the argument you’re going to make actually is. Only then can you start writing the structure for an essay that builds up to your overall conclusion.
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Essays are a chance for you to show off how widely read you are, so make sure you quote other people’s opinions, and phd thesis in computer networks original sources, on what you’re writing about. For example, if you were to write a history essay on early religious practices in faire une introduction dans Britain, you could quote original texts on that topic (such as Bede’s Ecclesiastical History of the English People ) and also mention what a range of modern scholars have to say about the topic. Contrasting views should be sought; it’s unlikely that everyone agrees on the topic, so show you’ve looked at all the possible angles. For each of the subjects you’re studying, start a page in a notebook for important people in that field, with a summary of when they lived and what their views are. That way, you’ll have something to refer to in computer networks, when you’re writing an essay and want to consult appropriate scholars or other writers whose opinions you might wish to include. Don’t quote too much; mix citations with your own opinions so that it doesn’t look as though you have to hide behind other people’s words. It’s fine to disagree with a scholar you quote, provided you can give evidence and reasoning for doing so. This shows that you have thought about it and made your own mind up, rather than blindly accepting what that scholar has said; this demonstrates strong critical reasoning skills, one of the hallmarks of in computer networks brilliant students. 6. Phd Thesis! Syntax, punctuation and tone of voice. Be honest: do you find your tone of voice interesting? You may not consciously realise it when you’re reading, but sophisticated sentence structures make the world of difference to how intelligent you sound.
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Lactoferrin receptor mediates apo- but not holo-lactoferrin internalization via clathrin-mediated endocytosis in trophoblasts. LfR [Lf (lactoferrin) receptor] is phd thesis networks, expressed in most mammalian tissues, including placental trophoblasts, and is presumed to mediate the internalization of Lf. However, the comment dans, physiological significance of 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. As Lf functionality is in computer, related to the presence (holo-Lf) or absence (apo-Lf) of iron within the i live, 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 networks ligand blotting confirmed that Lf binds to a LfR of cover for waitress with ?105 kDa. Apo-Lf treatment significantly increased LfR abundance at phd thesis in computer 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 hypertonic medium (0.4 M sucrose) significantly inhibited apo-Lf internalization. In summary, our data demonstrate that apo- but not holo-Lf is internalized by LfR and suggest that, following internalization via LfR, apo-Lf plays a role in CT invasiveness by une introduction dans une dissertation, 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 phd thesis in computer, 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 phd thesis networks a LfR [Lf (lactoferrin) receptor] was first identified in in computer networks, human small intestine . It presumably facilitates the uptake of its protein ligand Lf, which is present in on the self i, milk and mucosal secretions .
Therefore, kinetic studies on LfR have primarily been conducted in in computer networks, small intestine [3–6] and cultured enterocytes . Dans! We have previously cloned the LfR gene (GenBank® accession number NM_017625)  and determined that the intestinal LfR is an ?110 kDa trimeric (?37 kDa subunits), glycosylated protein . Networks! However, LfR is also expressed in non-intestinal tissues, such as heart, liver, spleen, thymus and role skeletal muscle [3,8,9]. Recently, we detected LfR expression in mouse placenta associated with trophoblast cells , suggesting that LfR plays a role in in computer networks, this highly specialized cell type. Trophoblasts are divided into two subtypes; CTs (cytotrophoblasts) and STs (syncytiotrophoblasts), which are both maintained throughout pregnancy. The primary function of CTs is invasion into comment dans une dissertation, the uterine epithelium to secure successful implantation , whereas STs further implantation progression by establishing the networks, nutrient and gas exchange barrier (maternal–fetal interface) and secreting hormones such as progesterone which are essential for successful pregnancy maintenance . The invasive capacity of CTs relies on the secretion of phd thesis in computer MMPs (matrix metalloproteinases), which function to degrade the surrounding ECM (extracellular matrix). Several disorders of pregnancy, including pre-eclampsia and phd thesis in computer spontaneous abortion, are believed to relate to ineffective CT proliferation, migration and statement at the invasion into the uterine endothelium . Lf has been attributed with multifunctional properties, including increased cell proliferation [12–16] and changes in networks, cell morphology through activation of MMP transcription .
The pleuripotency of Lf may reflect the une introduction, dramatic conformational change associated with iron binding and release [18,19] as holo-Lf (iron saturated Lf) and networks apo-Lf (iron free Lf) are known to adopt “closed” and “open” forms respectively [18,20]. For example, holo-Lf increases but apo-Lf decreases cell proliferation in human enterocytes (Caco-2 cells)  and essay self who am i while apo-Lf stimulates microvessel proliferation in rats, holo-Lf is ineffective . Currently, the mechanisms through which apo- and holo-Lf exert their differential functions are unknown. Herein, we hypothesized that the opposing functionality of apo- and holo-Lf reflects differences in cellular internalization mechanisms mediated via LfR. In order to phd thesis in computer networks address this, we utilized the human placental CT cell line BeWo and cover for waitress determined: (1) the effects of apo- and holo-Lf on MMP-2 activity, the major MMP in trophoblasts; (2) the ability of the LfR to in computer networks facilitate apo- and faire dans une dissertation holo-Lf uptake and internalization; and (3) the subsequent intracellular compartmentalization of LfR following Lf uptake. We also determined that apo- but not holo-Lf stimulates MMP-2 activity, that apo-Lf is internalized by LfR in phd thesis, a clathrin-mediated process and that internalized LfR associates with the early endosome/recycling compartment. Letter For Waitress With No Experience! 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 phd thesis networks, 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 37 °C in a humidified atmosphere with 5% CO 2 until 90% confluent. For experiments, cells were sub-cultured at a ratio of for waitress with 1:3 and cultured until confluent. Lf was isolated from human milk as previously described with slight modifications . Casein was precipitated from pooled human milk with 0.06 M CaCl 2 , pH 4.6, for 1 h at room temperature (25 °C) with constant stirring and centrifuged at 8600 g for in computer networks, 20 min at essay on the self 4 °C. Phd Thesis! The whey fraction was collected, centrifuged at 8600 g for 20 min at 4 °C and then mixed with heparin–Sepharose Fast Flow resin (GE Health-Amersham Biosciences) overnight at 4 °C. The whey/resin mixture was centrifuged at for waitress with no experience 800 g for 5 min at 4 °C, the supernatant discarded and networks 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 fractions were collected. Letter For Waitress No Experience! Lf-containing fractions were identified following Coomassie Brilliant Blue staining of SDS/PAGE (12%) gels. Phd Thesis Networks! Samples were pooled and concentrated in YM-50 centrifugation tubes (Millipore Centriplus, Fisher Scientific) at 3000 g for 90 min at responsibility 4 °C. The protein concentration of purified Lf was determined spectrophotometrically ( A 280 ) as previously established . Apo-Lf was prepared as previously described . Holo-Lf was prepared with FeSO 4 (Lf/Fe, 1:2 molar ratio) in phd thesis, sodium bicarbonate (Fe/HCO 3 , 1:1 molar ratio) for dual of women, 1 h at room temperature (25 °C). Unincorporated Fe was removed with YM-50 microcentrifuge tubes at 12000 g at 4 °C and phd thesis in computer stored at ?20 °C. Comment Dans Une Dissertation! Fe saturation was determined spectrophotometrically by the A 460 / 280 ratio (22) .
To determine effects of phd thesis networks apo- and faire une dissertation holo-Lf on MMP-2 activity, cells were seeded in phd thesis in computer, 24-well plates in complete medium until cells were 90–95% confluent. 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 one third of the original volume using YM-50 microcentrifugation tubes (Millipore Centriplus) at 5000 g for 20 min at 4 °C. Protein concentration of conditioned media was determined using the Bradford assay (Bio-Rad) and proteins (20 ?g) were electrophoresed by role, SDS/PAGE (7.5% gel containing 1 mg/ml gelatin). Following electrophoresis, the gels were washed twice in 2.5% Triton X-100 in phd thesis in computer networks, PBS for 15 min to essays remove SDS and phd thesis incubated in for waitress no experience, 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 in computer networks destained with 10% acetic acid and 40% methanol in water. Responsibility! Absence of stain (representative of proteolytic degradation) reflects MMP-2 enzymatic activity.
The relative densities of 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 . Excess free iodine was removed with a PD-10 desalting column (GE Health-Amersham Biosciences). Networks! To determine Lf uptake, cells were cultured in 24-well dishes in growth medium until confluent. Essays! Cells were incubated with 125 I-labelled apo- or holo-Lf (10000–100000 c.p.m.) in SFM (serum-free medium) for 1 h at in computer 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 radioactivity was quantified in both medium and solubilized cells in a gamma counter. Affinity purification and about responsibility validation of LfR antibody. LfR peptide (SSSPSLPRSCKEIKDE-Cys) was produced (Genemed Synthesis, South San Francisco, CA, U.S.A.) and phd thesis in computer networks conjugated (2 mg) to an affinity-purification column (Sulfolink purification kit; Pierce Biotechnology).
LfR antibody was purified from letter with no experience, rabbit antiserum according to the manufacturer's instructions. To demonstrate antibody specificity, affinity-purified LfR antibody (1 ?g/ml) was pre-incubated with LfR peptide (1 mg/ml) in 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 networks, 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 the Bradford method and dual role essays cell lysates (50–100 ?g protein) were resolved by electrophoresis. To determine Lf binding, cell proteins were resolved by non-reducing SDS/PAGE at 80 V for 4 h and phd thesis 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 dual of women essays bound ligand was visualized by autoradiography. For immunoblotting, cell extracts were elecrophoresed by SDS/PAGE (10% gel, 200 V for in computer networks, 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 essay on the self who am i, PBS-T.
Membranes were washed in PBS-T for 30 min, visualized with Super Femto Detection Chemiluminescence Reagent (Pierce Biotechnology) and exposed to autoradiography film. Relative band density was quantified using the phd thesis in computer, Chemi-doc Gel Quantification System (Bio-Rad). Transient transfection with LfR siRNA (small interfering RNA) Cells were seeded on of women, 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. Phd Thesis In Computer! 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 SFM at 37 °C. Lf uptake was measured after 30 min as described above. To verify LfR suppression, transfected cells were scraped into 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 letter with immunoblotted as described above. Biotin-labelling of cell surface proteins.
To determine LfR abundance at the plasma membrane, cells were seeded on in computer, to 100-mm-diameter dishes and cultured until confluent. 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 statement for why at the 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 20 min at 4 °C. Phd Thesis In Computer! The supernatant was discarded and the pellet was resuspended in lysis buffer with 0.2% SDS and une dissertation incubated at 60 °C for 5 min. As the samples cooled, 1% Triton X-100 was added, the samples were re-sonicated and centrifuged at phd thesis in computer 100000 g for 20 min at essays 4 °C to pellet any insoluble material. The supernatant was collected and phd thesis in computer networks incubated with 300 ?l of letter slurry (1:1) of immobilized avidin beads (UltraLink Immunobilized Monomeric Avidin; Pierce) for 1 h at in computer room temperature (25 °C) with constant rotation. To collect proteins bound to the resin, the samples were centrifuged at 2300 g for 1 min. Une Introduction! The supernatant was discarded and the beads were rinsed with PBS and centrifuged for phd thesis networks, 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 about responsibility 90 °C for 5 min.
The samples were centrifuged at 2300 g for 1 min and biotinylated proteins were resolved by networks, 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 statement for why at the p.o, a GPI-anchored protein, cells were seeded on to 24-well plates and pretreated with Pi-PLC (1 unit/ml; Invitrogen) in SFM for 1 h at phd thesis in computer 37 °C to cleave GPI-anchored proteins. To verify the effects of Pi-PLC on LfR abundance at the plasma membrane, cell surface proteins were biotinylated and letter with immunoblotted for LfR as described above. To determine the effects of Pi-PLC on Lf uptake, cells were rinsed with PBS, fresh SFM was added and 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 phd thesis, efficacy of the enzyme, spent medium was collected and concentrated to for why one third of the phd thesis networks, original volume using YM-50 microcentrifugation tubes (Millipore Centriplus) at 5000 g for 20 min at essays about responsibility 4 °C and 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). Networks! Cells were pre-treated in hypertonic medium for 30 min and incubated with 125 I-labelled apo-Lf in SFM at 37 °C in the presence of 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 required for comment une dissertation, 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 phd thesis 37 °C. Cells were either collected for 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 to glass coverslips, cultured for 3–4 days in growth medium and incubated with apo-Lf (where indicated, 200 ?g/ml) in cover no experience, SFM for up to 1 h at 37 °C. 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 phd thesis in computer networks, 5 min. To eliminate non-specific binding, cells were incubated in blocking buffer (5% goat serum in PBS) for essays about, 30 min followed by incubation with affinity-purified LfR antibody (0.8 ?g/ml) in networks, PBS containing 5% goat serum, 1% BSA and for waitress no experience 0.2% Triton X-100 for phd thesis in computer networks, 1 h. Cells were rinsed with PBS and LfR was detected with Alexa 488-conjugated-anti-rabbit IgG (1 ?g/ml; Molecular Probes) in PBS containing 0.2% Triton X-100 and essay who am 1% BSA for phd thesis in computer, 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 late endosomes; 1:500, Abcam) and detected with Alexa 568-conjugated-anti-mouse IgG (1 ?g/ml; Molecular Probes). After several rinses, coverslips were mounted with ProLong Gold (Molecular Probes) and sealed with nail polish. Immunofluorescence imaging was performed using an about, Olympus BX50WI microscope with UPlanApo ?100 magnification under an in computer networks, oil-immersion lens [NA (numerical aperture), 1.00], and digital images were captured using the Bio-Rad Radiance 2100 confocal system with LaserSharp2000 software, version 4.1 (Bio-Rad). Results are presented as means±S.D. of triplicate samples from 2–3 separate experiments. Statistical comparisons were performed using Student's t test or one-way ANOVA (Prism Graph Pad) and significance was set at P 0.05.
Active form of MMP-2 is increased by apo- but not holo-Lf. Secretion of MMP-2 plays a key role in CT invasion, thus we determined effects of Lf and Lf conformation on cover letter for waitress no experience, MMP-2 activity in BeWo cells. Using gelatin zymography, our data demonstrated that apo-Lf significantly increased (20%) MMP-2 activity compared to phd thesis networks untreated cells (Figure 1). 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 cover letter for waitress no experience 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 holo-Lf treated cells (lower panel). In Computer Networks! Data shown represent MMP-2 activity as a percentage of untreated control cells (mean±S.D.) for two independent experiments ( n =4). * P 0.05. LfR facilitates uptake of 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. Essays Responsibility! 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 ?35 and ?105 kDa in BeWo cells (Figure 3). Ligand binding plays a critical role in receptor function, and in computer ligand binding established that radiolabelled apo-Lf bound to two LfR proteins at ?75 and ?105 kDa (Figure 3), but not to the 35 kDa protein. We speculate that the 35, 75 and 105 kDa moieties represent LfR monomers, homo-dimers and -trimers, respectively. The ability of LfR to dual of women internalize apo-Lf is dependent on its cellular localization and we showed that the 105 kDa LfR was localized to phd thesis networks the plasma membrane using cell surface biotinylation followed by immunoblotting (Figure 4A). In contrast to what was reported for thesis for why i live at the, the intestinal LfR , trophoblast LfR does not appear to be GPI-anchored as demonstrated by networks, internalization of Lf following GPI-anchored protein cleavage with Pi-PLC (Figure 4B).
Confirmation of Pi-PLC cleavage was verified as Pi-PLC increased ALP abundance in letter, spent medium compared to phd thesis in computer networks untreated cells (Figure 4C). Confluent cells were treated with 125 I-labelled apo- or 125 I-labelled holo-Lf and letter no experience Lf uptake was determined. Representative graph of uptake after 1 h illustrates that significantly less holo-Lf was internalized when compared to 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 networks, 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. Phd Thesis In Computer! The LfR antibody detected specific proteins at phd thesis in computer ?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 on the i non-denaturing conditions, and incubated with 125 I-labelled apo-Lf. 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 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 protein±S.D. for two independent experiments ( n =4). In Computer Networks! ( 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 Lf uptake, LfR expression was attenuated in cells transfected with LfR siRNA (Figure 5A). Comment Dans! We demonstrated that reduced LfR abundance resulted in 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 phd thesis in computer networks holo-Lf in BeWo cells is not predominantly LfR-mediated. Due to our finding of phd thesis in computer limited LfR-mediated holo-Lf uptake and lack of an effect of 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 mock-transfected cells. ( B ) Lf uptake was measured using 125 I-labelled apo- or 125 I-labelled holo-Lf in networks, cells transfected with LfR siRNA or mock-transfected controls. Essay On The Self! Data represent the phd thesis, means±S.D. Thesis For Why P.o! ( n =2–3) of triplicate experiments. * P 0.05, ** P 0.01, as analysed by Student's t test. LfR internalization is a clathrin-dependent process and phd thesis in computer 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. Faire Une Introduction Dans! 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 in computer networks changes in total LfR abundance (Figure 6B). This suggests that apo-Lf may signal the faire une introduction dans une dissertation, recruitment of phd thesis in computer LfR to the plasma membrane or that perhaps Lf binding to LfR transiently interferes with its internalization. Confluent cells were treated with or without apo-Lf and LfR surface expression was assessed. ( A ) Representative immunoblot of crude membrane proteins from cells treated with apo-Lf or untreated and detected with LfR antibody. On The Who Am I! 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 ). * P 0.001, as analysed by Student's t test. ?-actin was used as a loading control. Data represent means±S.D. ( n =3) of phd thesis 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 inhibit clathrin-dependent endocytosis, which resulted in a robust inhibition of i apo-Lf internalization (40%) compared with untreated cells, with values of 2.4±0.25 and 5.7±0.42 pmol/mg protein±S.D. for treated and control cells respectively; n=5–8 per phd thesis in computer networks group, * P 0.001. To confirm the mechanism by cover letter for waitress no experience, which LfR is endocytosed, we used confocal microscopy to networks 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 phd thesis in computer, yellow pixilation in the merged images (Figure 7C).
As internalization is ligand-stimulated, cells were treated with apo-Lf, and indirect immunofluorescence was used to visualize specific sub-cellular compartments within which LfR was associated. Phd Thesis In Computer Networks! Following ligand stimulation, we demonstrated partial co-localization of LfR and the early endosome marker EEA1 in vesicular/endosomal-like structures (Figures 7D–7F). As co-localization between EEA1 and LfR was not exclusive, we sought to examine other sub-cellular compartments containing LfR. Comment Faire Une Introduction Dans Une Dissertation! Clathrin-mediated endocytosis is one mechanism cells utilize to control cell surface receptor abundance and turnover via recycling. To investigate whether LfR trafficking from the plasma membrane potentially includes endocytic recycling, we used the phd thesis in computer, indirect immunofluorescence of in computer TfR as a marker for phd thesis, recycling endosomes. Self I! We showed partial co-localization of LfR and phd thesis in computer TfR within distinct internalized vesicles in thesis statement at the p.o, cells following ligand stimulation (Figures 7G–7I).
No co-localization between LfR and M6PR (a late endosome marker) and minimal co-localization with LAMP1 (a lysosome marker) was detected (results not shown). These results indicate that, in 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. Phd Thesis In Computer! Cells were fixed with paraformaldehyde and imaged for in computer networks, LfR (green) and networks sub-cellular markers (red); clathrin ( A and C ), EEA1 ( D and F ) and TfR ( G and I ). Clathrin ( A ) and of women 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 early endosomal marker EEA1 ( D ) and LfR ( E ) was detected (yellow, F ) and phd thesis networks treatment of cells with apo-Lf clearly resulted in vesicularization of thesis for why LfR.
Furthermore, co-localization with the phd thesis, recycling endosome marker TfR ( G ) and LfR ( H ) was detected (yellow, I ) following treatment of for why i live cells with apo-Lf. Finally, to determine if increased abundance of LfR at phd thesis networks the plasma membrane in cover letter with no experience, response to apo-Lf stimulation reflects changes in in computer, intracellular LfR recycling, we blocked de novo protein synthesis with Cyhex and measured apo-Lf uptake. Letter For Waitress No Experience! Inhibition of phd thesis in computer new receptor synthesis did not diminish apo-Lf uptake, indicating that LfR is recycled. In fact, apo-Lf uptake was significantly higher in for waitress with, 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 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 phd thesis LfR abundance was assessed compared with untreated cells. Cover For Waitress With! ( A ) Apo-Lf uptake from cells significantly increased in networks, the presence of cycloheximide (Lf+Cyhex) after 3 h compared with untreated cells (Lf). Data represent means±S.D. for two independent experiments ( n =3–4), and * P 0.05 compared with untreated cells, as analysed by Student's t test. ( B ) Representative immunoblot of total protein extract isolated from cells pre-treated with Cyhex then incubated with apo-Lf for 3 and 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 . Essays About! Our previous detection of LfR along the maternal and fetal interface in the placenta implicates LfR in CT function [8,9,27].
The aim of the present study was to explore the function of phd thesis networks Lf in the placenta as mediated via LfRs using a CT cell model. While primarily regarded as a component of milk , Lf is also found in decidua (9–95 ?g/g), amniotic fluid (2–37 ?g/g) and chorion (2–26 ?g/g) , and is secreted directly from uterine epithelia [29,30]. This suggests that Lf secreted from the dans une dissertation, uterine endothelium may regulate CT function in a paracrine manner. Results from this study documented an in computer, enhancing effect of apo- but not holo-Lf on MMP-2 activity in on the who am, CTs, suggesting a regulatory role for apo-Lf in CT invasivity as this process is dependent upon the secretion and activation of MMPs. There is precedence for effects of networks Lf on MMP regulation as over-expression of essay self who am i Lf activates MMP-1 expression through binding to AP-1 promoter elements and networks through stimulation of the p38 MAPK (mitogen activated protein kinase)-dependant pathway in mouse fibroblasts and about responsibility human COS-1 cells . In Computer! 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 . Cover Letter With! In support of the in computer networks, concept of Lf acting as a transcription factor, a nuclear localization signal within the N-terminal region of the Lf molecule (amino acids 1–5) has been previously identified , and Lf binds to 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 MMP-2 protein expression . The ability of Lf to activate p38 MAPK has been previously shown in statement i live p.o, other cell types [16,35], providing support for phd thesis in computer, a direct role of apo-Lf on CT invasivity.
The effect of apo- but not holo-Lf on phd thesis in computer, MMP-2 activity probably reflects that distinct functions are associated with either apo- or holo-Lf. In contrast to reports in rat hepatocytes , CTs bound and internalized both apo- and phd thesis in computer 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 phd thesis in computer networks this molecule for CT-specific functions such as invasivity, while precise Lf-dependant functions in 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 , which have been postulated to determine ligand binding and in computer receptor interactions.
It is about responsibility, possible that iron-saturation, which alters Lf conformation, may result in LfR inaccessibility to these binding motifs. While this is not to imply that holo-Lf binding and uptake does not elicit cellular events, it does not appear that these effects are mediated via LfR internalization in phd thesis networks, CTs. Letter No Experience! 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 LfR, we sought to characterize the placental LfR more extensively. It has previously been reported that intestinal LfR is phd thesis, a GPI-anchored protein . Contrary to this observation, we determined that, in dual role of women essays, CTs, the LfR receptor is not a GPI-anchored protein and is in fact internalized via clathrin-mediated endocytosis following ligand binding. Our observations in phd thesis networks, trophoblasts are consistent with previous reports in other cell types such as hepatocytes, where inhibition of on the self i clathrin-mediated endocytosis by hypertonic medium  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 phd thesis invasive cell type, whereas enterocytes are terminally differentiated polarized cells involved in vectorial nutrient transport. 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 interaction with adaptor proteins, such as AP2 . Cover With No Experience! To identify potential AP2-interacting motifs within the LfR sequence we used a proteomics server (www.elm.eu.org) and in computer networks indeed two tyrosine-sorting motifs were identified at amino acids 120–123 and 129–142. This further suggests that a portion of LfR protein does reside intracellularly and that one or more of 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 . Phd Thesis In Computer! Recently, Tsuji et al.  examined the structure of phd thesis human LfR in greater detail and letter determined that it is an phd thesis in computer, 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 for why i live, abundance of the phd thesis networks, trimer was always greater that the monomer.
For some reason, we were unable to extensively reduce the trimeric form to monomers even when using dithiothreitol and about responsibility iodoacetic acid (V. Lopez, unpublished observations). Interestingly, we did not detect the in computer networks, dimeric form of LfR under reducing conditions, which is similar to the findings of Tsuji et al. . We further demonstrated that apo-Lf binds to on the who am 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 . We speculate that the trimeric form is an unusually stabilized form, mediated by phd thesis in computer, several intracellular disulfide bonds and possibly also hydrophobic interactions, whereas the dimeric form is less stable and can be reduced to the monomeric form. In contrast, a 45 kDa calcium-dependent LfR has been reported in rat hepatocytes ; however, it is unknown how this LfR relates specifically to Lf function as rats do not express Lf. It is well established that cell signalling and cellular homoeostasis are altered by increasing or decreasing cell surface receptor abundance . As LfR abundance at the plasma membrane did not require de novo synthesis, our data suggest that recycling of LfR to and from the plasma membrane occurs. Cover Letter For Waitress With No Experience! Endocytic recycling requires internalization of phd thesis cell surface receptors and their ligands within clathrin coated pits , with the interaction between transferrin and TfR being the essay self who am i, classic example of phd thesis networks this process . Receptor recycling is one strategy by which cells maintain a mobilizable receptor pool to self quickly traffic to the cell surface in response to physiological stimuli, to allow cells to redistribute cell-surface molecules to specific cell locations or compartments undergoing a physiological activity .
We speculate that Lf may either stimulate the recruitment of LfR to the plasma membrane, or that perhaps Lf binding to the receptor results in phd thesis networks, transient accumulation of une dissertation LfR prior to internalization via clathrin-mediated mechanisms. We propose that CTs may require Lf/LfR internalization to networks rapidly facilitate implantation, allowing for optimal embryo-fetal development. Further studies are needed to address this issue. Collectively, the cover no experience, results from this study demonstrated a specific role for in computer, apo-Lf but not holo-Lf in phd thesis, regulating MMP-2 activity and in computer networks thus potentially endothelial invasion in role of women, CTs. Importantly, we have determined that apo- but not holo-Lf is internalized via LfR, which occurs through a clathrin-dependent pathway in CTs suggesting a mechanistic and functional difference between apo- and holo-Lf. These results may help to explain the pleuripotent and cell-specific functions which have been associated with Lf.
The authors would like to phd thesis in computer networks express their gratitude to comment une introduction une dissertation Dr. Lisa L. Gough for critical reading of the 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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Should i include collegiate sports in my resume? And if so where? It depends how much professional experience you have. If you are a recent college grad, then it is acceptable to include on your resume. Good luck on the job hunt!
Good luck on the job hunt! If the jobs are relevant to phd thesis networks, the ones you are applying for, then you can go as far back as you like. With regards to your military experience, check out thesis statement for why at the p.o our military to civilian resume guide: https://resumegenius.com/blog/go-shooting-guns-coffee-runs. Any of the templates in library 2 would be suitable for manufacturing careers. Best of phd thesis in computer networks luck! I’ve worked in the same industry for the past 13 years. Multiple employers with jobs lasting two to three years each. The jobs have been similar, so the experience looks a bit repetitive. I need to find a template that highlights my experience without getting bogged down in the chronology, Any suggestions? It provides ample space for your professional experience, while also highlighting your top qualifications.
Good luck on the job hunt! hi resume genius.. i need template resume that suitable for trainer and coach.. can u suggest to me with template is suitable.. Statement I Live? #128578; I had a job for 7 years and during that time I wore many hats, Executive Admin, Purchasing, Vendor Management, Project Coordination, etc. How would I write that on my resume? Perhaps the in computer, Company name and then all the related roles under that and the times I did those jobs? I was always the Executive Admin, but I did other jobs during that period. Yes, your suggestion is phd thesis networks correct. Start with the networks, company name and included the who am i, related jobs with their own bullet points underneath.
Good luck! Consider trying the ‘Job Hopper’ or the ‘Executive.’ They should able to fit all your jobs nicely. Ive never had a job so what should I use? Most of the phd thesis in computer networks, templates above would suit your situation, but we suggest trying the dual of women, Career Changer template because it emphasizes skills over the dates of phd thesis in computer your professional experience. (https://resumegenius.com/resume-templates/career-level-life-situation-templates#career-changer-templates) Best of luck! We suggest using the ‘Gatsby’ Template. Good luck with grad school! As far as style, we suggest our ‘Professional’ template. In terms of format, if you want to include your restaurant experience, then you might want to consider using a functional format:https://resumegenius.com/resume-formats/functional-samples-writing-guide. Hope this helps! We suggest using our ‘Entry-Level’ template. Essays About? Good luck with the internship!
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So i want to highlight the other things. Thanks #128578; Take a look at our Flight Attendant resume sample: https://resumegenius.com/resume-samples/flight-attendant-resume-example You can download it and essay on the self who am i, input your own information. Which template would you recommend for a career in education? Check out our teacher resume samples: https://resumegenius.com/resume-samples/teacher-resume-example You can download them and input your own experience. Try using the ‘Freeman’ template.
Best of in computer networks luck on the promotion! Hi! What resume template would you recommend for a college freshman trying to apply for a competitive summer program with the USDA and South Dakota State University? Sound like the ‘Entry-Level’ template would be a good fit for about responsibility what you’re trying to phd thesis in computer, do. Good luck with the summer program.
Hi! Which resume template would you recommend for someone trying to tap into the finance and accounting market. Looking for an entry-level position. You should go with the ‘Entry-Level’ template. Good luck with the job hunt. I have worked 32+ years as a nurse, the last 4 years taking care of my elderly father and essays, online work. Now seeking to get back into in computer networks, the job market for phd thesis in computer extra income, not necessarily in the health field, just to phd thesis in computer networks, earn some income and socialize. Role Of Women? What resume do you suggest? Try the ‘Job Hopper’ template.
Good luck with your job search! Hi! What resume template would you recommend for a 9th grader trying to apply for in computer a doctor (any)?? Apparently, resume making and une introduction, interviewing is our project for the fourth quarter this year. I couldn’t find any clear examples on the web, and I was hoping you could help me out with what template I should use..
Try using the phd thesis, ‘Elegant 2.0’ template. Thesis I Live P.o? Good luck on phd thesis in computer networks your project. Yes, if you click the View all Resume Designs button and role essays, click the download link for phd thesis in computer networks the template pack of your choice. If you’ve never written a resume before, I’d recommend checking out our “How to Write a Resume” guide to get a clearer idea (it’s much more comprehensive than any answer I can give here). https://resumegenius.com/how-to-write-a-resume. Hit us up with any follow-up questions after giving that a read we’ll see if we can help further!
Good luck! Hey there Margaret, In order to letter, best understand which template works, it’s a good idea to check out which resume format fits your particular needs; then you can take it from there. In Computer? https://resumegenius.com/resume-formats. All of the templates were created by about responsibility professional resume writers, so it’s hard to go wrong with any of them — it just depends on your preference. Good luck! It really depends on what job you’re applying for.
Since you have substantial work experience, try quantifying that in your resume (think: any numbers that a hiring manager can look at and better understand what you accomplished during your time working there). Check out this page and choose the one you find most fitting, that should be a good start: https://resumegenius.com/resume-formats. Good luck on the job hunt! Hey there hbil036, This way, you can focus on your skills qualifications critical to the job application. As an aside, you may want to look into whether you’re qualified to get back into accounting after that many years outside of the field. I understand that some regulations and rules change over the years — it may just be a matter of taking a test or updating your certifications, but I’m not certain. If that doesn’t seem to be a problem then go with the functional resume for sure. Good luck on the job hunt! If you are lacking in major experience, I’d recommend using a reverse chronological format for your resume. Our “Classic” template on this page should do the trick: https://resumegenius.com/resume-templates/ Good luck at phd thesis in computer, the job fair!
I recommend you first check out our internship resume sample page: https://resumegenius.com/resume-samples/internship-resume-example. Dual Role Of Women? Afterwards, feel free to choose any format – just use a comprehensive education section instead of a professional experience section, and you should be good. Good luck landing that internship! Share Free Downloadable Resume Templates Our code geeks and HR experts are proud to phd thesis in computer networks, introduce our new Free Resume Builder software to help you land more interviews in today’s competitive job market.
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5 simple steps to in computer networks create your first RESTful service. RESTful services are those which follow the REST (Representational State Transfer) architectural style. Before implementing your first RESTful service, lets first understand the concept behind it. As we know that WCF allows us to make calls and exchange messages using SOAP over a variety of protocols i.e. HTTP, TCP, Named Pipes and MSMQ etc. For Waitress With No Experience! In a scenario, if we are using SOAP over HTTP, we are just utilizing HTTP as a transport. But HTTP is phd thesis in computer much more than just a transport. So, when we talk about REST architectural style, it dictates that “Instead of using complex mechanisms like CORBA, RPC or SOAP for communication, simply HTTP should be used for making calls”. RESTful architecture use HTTP for all CRUD operations like (Read/Create/Update/Delete) using simple HTTP verbs like (GET, POST, PUT, and DELETE).It’s simple as well as lightweight.
For the sake of simplicity, I am going to dual role of women implement only a GET request for which service will return certain types of data (i.e. Product data) in phd thesis, XML format. Create a WCF Service Project. Preparing the data (e.g. Product) to return Creating Service Contract Implementing Service Configure Service and Behavior. You can download the complete source code for this article here. Open Visual Studio.
From File - New Project . About Responsibility! Select WCF from left and create a new WCF Service Application . Phd Thesis In Computer! 2. Preparing the data to return. Essays About Responsibility! Now add a class to in computer networks newly created project. Name it to Products.cs. 3. Creating Service Contract. It will add contract as well as service file to project. Phd Thesis! Following is the code for service contract i.e.
IProductRESTService.cs. Phd Thesis In Computer Networks! IProductRESTService contains only dual of women essays, one method i.e. Phd Thesis In Computer! GetProductList. Important points to who am i understand about this method is WebInvoke attribute parameters. Method = #8220;GET#8221;, represents an HTTP GET request. ResponseFormat = WebMessageFormat.Xml, response format will be XML here but we can return JSON as well by changing its value to in computer networks WebMessageFormat.json. BodyStyle = WebMessageBodyStyle.Bare, indicates neither the request and nor response are wrapped. Other possible values for cover letter no experience, BodyStyle are Wrapped , WrappedRequest , WrappedResponse . UriTemplate = #8220;GetProductList/#8221;, it has two parts, URL path and query. Don#8217;t forget to add using System.ServiceModel.Web at phd thesis in computer, top. 4. Statement For Why! Implementing RESTful Service.
In this step we are going to implement the service. Only one method GetProductList is defined in the contract, so implementing service class will be as follows: 5. Configure Service and Behavior. The last step is to configure the service and phd thesis its behaviors using the for why i live, configuration file. Following is the complete ServiceModel configuration settings. Right click ProductRESTService.svc file and click #8220; View in Browser #8220;. You will see the following screen, that means service is fine. An important point to consider here is that in Service Behavior Configuration , we have setted httpGetEnabled=#8221;true#8221; for serviceMetadata that#8217;s why we are getting above service screen with wsdl option. Phd Thesis! This is what we do normally for SOAP-based services.
REST-based services only on the self who am, uses HTTP verbs on a resource, so we can disable WSDL in this case by phd thesis in computer, simply setting httpGetEnabled=#8221;false#8221; . Now if we run the service again, we will get the following screen. Just modify the URL in thesis statement i live, browser and add #8220; GetProductList/ #8221; to phd thesis in computer it. So, this is the UriTemplete we defined as service contract method . Hopefully, this simple WCF tutorial will be helpful for the readers. To keep the of women essays, things simple, I restrict it to just getting records using HTTP GET verb. But you can find a complete implementation for phd thesis in computer networks, all CRUD (Create, Read, Update, Delete) operations using RESTful service in another WCF RESTful Service tutorial here. Top 10 Interview Questions and Answers Series: Imran Abdul Ghani is working as Software Developer(Senior) with extensive knowledge in Web development technologies especially C#, ASP.NET, MVC, WCF, Web API, ADO.NET Entity Framework, jQuery etc. He has several years of cover, experience in designing/developing enterprise level applications.
He is Microsoft Certified Solution Developer for .NET (MCSD.NET) since 2005. You can reach his blogging at www.webdevelopmenthelp.net, www.topwcftutorials.net, and www.sharepointfordummies.net. public static string PostServiceRequest(string url, string JSONFilter) string result = string.Empty; var httpWebRequest = (HttpWebRequest)WebRequest.Create(url); using (var streamWriter = new StreamWriter(httpWebRequest.GetRequestStream())) var httpResponse = (HttpWebResponse)httpWebRequest.GetResponse(); using (var streamReader = new StreamReader(httpResponse.GetResponseStream())) save time thanks. worked right away. thanks. Thanks for phd thesis in computer, this Tutorial , how can i pass a parameter before i can get the comment une introduction, data in this tutoral and how can i make my Restful WCF Service. Explaination is good here too, but Code at this link works fine with same easy steps as here but with working result. Why does my REST service created throught this method has a wsdl ? I was under the in computer, impression that RESt services wouldnt have a wsdl . Role Essays! The Url for REST works fine and phd thesis networks I#8217;m able to in computer networks see the XML output . Yet when i put the url with ?wsdl it shows up #8230;. Running into a problem with: public List ProductList. The error states: Using the generic type #8216;System.Collections.Generic.List#8217; requires 1 Type argument. I have updated this WCF Tutorial to resolve the issue for networks, certain versions. You can follow the steps to have a clean implementation for WCF RESTful service.
Hopefully, I#8217;ll upload source code for the implementation shortly, so you will have a Visual Studio solution with complete source of the project. Thanks for your valuable feedback. For Why I Live P.o! how can i do this using xml data, pls help me. Can you please elaborate your question? As it#8217;s already using xml as response to render XML data. Please see the service contract as: Yes, How can i pass the xml data to service in PUT or POST Method which means RequestFormat = WebMessageFormat.Xml. I know how to call the service using jquery ajax call and pass the json datas, but don#8217;t know how to pass the xml values. Is it possible to phd thesis in computer networks pass the xml datas to in computer service using jquery or any other language should i use ? yes it#8217;s possible with following piece of sample code. string postData = SampleXml.ToString(); // convert xmlstring to byte using ascii encoding. In Computer Networks! byte data = encoding.GetBytes(postData); // declare httpwebrequet wrt url defined above.
HttpWebRequest webrequest = (HttpWebRequest)WebRequest.Create(url); // set method as post. // set content type. // set content length. // get stream data out of webrequest object. Stream newStream = webrequest.GetRequestStream(); newStream.Write(data, 0, data.Length); // declare read response from service. Comment Faire Une Introduction Dans Une Dissertation! HttpWebResponse webresponse = (HttpWebResponse)webrequest.GetResponse(); This answer is available at phd thesis networks, following URL. Thank you for self who am i, your valuable answer, i have already tried this code, but still i m getting 400 bad request error.. Phd Thesis In Computer! public List ProductList. Can you please explain what doesn#8217;t work for you. What error you#8217;re getting? Was this question answered? It#8217;s resolved now and updated.
Give me one more table class like Order, Combining both Products and Orders classes data want to get, please give that example. I#8217;ll do the une introduction dans une dissertation, above mentioned in later tutorial here. We come up with a complete example. Please revisit or just register by email. Create a class as below format: public class PostAlbumListRequest. public List Album. public User User. public string UserToken. In Computer Networks! [WebInvoke(Method = #8220;POST#8221;, UriTemplate = #8220;PostAlbum#8221;, RequestFormat = WebMessageFormat.Json, ResponseFormat = WebMessageFormat.Json)] PostAlbumResponse PostAlbum(PostAlbumRequest request); success: function (result) Very helpful for WCF beginers! Using VS 2013 Express. Essay Self I! Created WCF Service Application project.
Let me clear one thing as I mentioned in post step 1. You will create a new project using template #8220;WCF Service Application#8221;. Later in Step 3, you right click your project and add a new #8220;WCF Service#8221; to it. In Computer Networks! I#8217;m using VS2012. The WCF tab when creating a new project has; WCF Service Library, WCF Service application, WCF workflow service application sysndication service libray. This is using the .net framework 4. WCF Service or WCF Service Application template are available. In Computer Networks! I am not sure which version of in computer, Visual Studio you are using#8230; Just use WebApi#8230; The code is missing. Essay On The Who Am I! public interface IProductRESTService. Phd Thesis In Computer Networks! Yeah, I missed that. Faire Une Introduction Dans Une Dissertation! thanks. Metadata publishing for phd thesis in computer, this service is comment faire currently disabled. Please verify you follow the step 5. Please confirm following in your configuration file.
Using WebApi is way much easier! WebAPI is another way of doing the same. You can find almost same in Web API at http://www.webdevelopmenthelp.net/2013/09/ASP-Web-API-Service.html. Networks! i agree that web api is easier but this can be helpful if your org dont have enough fundings to upgrade to iis 7 or iis 8.
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We may monitor and essays, record phone calls, e-mails, live chats, or other communications between You and our Customer Service representatives or other employees or representatives. Information We Collect Automatically. We automatically collect a variety of information associated with Your use of our Services. Phd Thesis In Computer. Each time You visit the Website, Personal Information is automatically gathered. In general, this information does not identify You personally. Examples of automatically collected personal information include, but are not limited to: IP address, Collection Date, Publisher Name, Connection Speed, Day of Week Time of Day (hour), Language settings, Country, City (relating to IP address, if available). For example, some of the ways we may automatically collect information include:
We may also obtain information about You from other sources. For example, We may receive credit information from third-party sources before initiating Your service. We may also purchase or obtain Personal Information (for example, e-mail lists, postal mail lists, demographic and marketing data) from others. HOW WE USE INFORMATION WE COLLECT ABOUT YOU. We use the information We collect for a variety of business purposes, such as: To provide and in computer networks, bill for Services You purchase; To deliver and confirm Services You obtain from us; To verify Your identity and maintain a record of Your transactions and interactions with us;
To provide customer services to You; To create, modify, improve, enhance, remove or fix our Services and their performance; To identify and suggest products or services that might interest You; To make internal business decisions about current and future Service offerings; To provide You customized user experiences, including personalized Services offerings; To protect our rights, interests, safety and property and essays, that of in computer our customers, service providers and other third parties; and. To comply with law or as required for legal purposes. We may use Personal Information for investigations or prevention of fraud or network abuse. We may use information we collect to contact You about cover letter for waitress, our and/or third-party products, services, and offers that We believe You may find of phd thesis in computer networks interest. We may contact You by telephone, postal mail, e-mail, or other methods.
You may see advertisements when You visit our Website. We may help advertisers better reach our customers by providing certain customer information, including geographic information, language preferences or demographic information obtained from other companies. This information is used by cover letter no experience advertisers to in computer networks determine which ads may be more relevant to You. However, we do not share Personal Information outside of our corporate family for letter advertising purposes without Your consent. WHEN WE SHARE INFORMATION COLLECTED ABOUT YOU. We do not sell, license, rent, or otherwise provide Your Personal Information to unaffiliated third-parties (parties outside our corporate family) without Your consent. We may, however, disclose Your information to unaffiliated third-parties as follows: With Your Consent. We may disclose Personal Information about You to third-parties with Your consent. We may obtain Your consent in phd thesis in computer writing; online, through “click-through” agreements; when You accept the terms of disclosures for certain Services; orally, when You interact with our customer service representatives.
We encourage You not to statement i live at the share Your password. If You provide Your user account password and/or security question responses to third parties they will have access to Your Personal Information when they access Your user account with Your account password. To Our Service Providers. We may disclose information to third-party vendors and phd thesis in computer networks, partners who complete transactions or perform services on our behalf (for example, credit/debit card processing, billing, customer service, auditing, and marketing). In a Business Transfer. We may sell, disclose, or transfer information about You as part of a corporate business transaction, such as a merger or acquisition, joint venture, corporate reorganization, financing, or sale of company assets, or in the unlikely event of insolvency, bankruptcy, or receivership, in which such information could be transferred to third-parties as a business asset in the transaction. For Legal Process Protection. We may disclose Personal Information, and other information about statement i live at the, You, or Your communications, where we have a good faith belief that access, use, preservation or disclosure of such information is reasonably necessary: to satisfy any applicable law, regulation, legal process or enforceable governmental request; to enforce or apply agreements, or initiate, render, bill, and collect for phd thesis services and essays about responsibility, products (including to collection agencies in in computer order to obtain payment for our products and services); to protect our rights or interests, or property or safety or that of others; in connection with claims, disputes, or litigation – in cover court or elsewhere; to facilitate or verify the appropriate calculation of taxes, fees, or other obligations; or. in an emergency situation. We may provide information that does not identify You personally to third-parties for in computer marketing, advertising or other purposes. HOW WE STORE AND PROTECT THE INFORMATION COLLECTED ABOUT YOU.
We may update this Policy at phd thesis networks any time to provide updates to or clarification of networks our practices. If we make changes we may provide You with additional notice (such as adding a statement to in computer networks the homepage of our Website or sending You a notification). Dual Role. You should refer to this Policy often for the latest information and the effective date of any changes. This web site is owned and phd thesis, operated by Viatta Business Ltd . A Partner is an individual who refers customers. A Referral is an individual who requests a service via the referral link given by a Partner. With the first order, a Referral acquires a 15% discount on the order, while a Partner receives $50 to i live at the p.o the Referral Balance.
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