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Most proteins are glycosylated. Mass spectrometry methods are used for mapping glycoprotein glycosylation and detailed glycan structural determination. This technology enables precise characterization of recombinant glycoproteins in the pharmaceutical industry and academic biomedicine.
This feature article is intended as a tutorial for scientists wishing to learn more about mass spectrometry applied to glycan and glycopeptide structure. It is not intended as a comprehensive review, and apologies are made to those whose work could not be referenced.
Despite the fact that glycosylation is energetically costly, all cells, in all kingdoms of life, are coated with glycoconjugates1. Not only are glycosylated molecules required for organismal success in an evolutionary context2, but they are required for myriad physiological functions. The functions of glycoconjugates cannot be understood fully using a linear model for information flow in biological systems; it is not possible to predict glycoconjugate structure and function from genomic information. Yet, the functions of glycoconjugates are a direct consequence of their non-template driven biosynthesis. Because the biosynthetic reactions that form glycoconjugate glycans are mediated by enzymatic reactions governed by complex factors including substrate availability and kinetics, glycan structures are heterogeneous as a rule. This results in a distribution of physico-chemical properties in the mature glycoconjugates and concomitant diversity of protein-binding properties. Glycan binding proteins have evolved with glycans and may be viewed as a lens for understanding the functions of glycosylation. Thus, the diversity of glycosylation is a mechanism whereby the binding interactions between a glycosylated protein and its ligands and/or receptors are modulated. In addition, glycosylation is dynamically regulated according to spatial and temporal factors in tissues; this drives the need for effective analytical methods for producing data to inform a clear understanding of glycobiology.
Polypeptide chains destined for the lysosome or for secretion are biosynthesized into the lumen of the endoplasmic reticulum. Many of these proteins are modified co-translationally by N-glycosylation using evolutionarily conserved machinery. These N-glycans help regulate protein folding by binding to calnexin and calreticulin. Through this quality control pathway, only correctly folded proteins are allowed to exit the endoplasmic reticulum.
As an example, consider antibody glycosylation. All antibodies are N-glycosylated at a conserved Asn residue in the Fc region. This glycosylation supports the three dimensional structure of the Fc region and is required for the effector functions whereby antibodies mediate their cytotoxic effects3. Monoclonal antibody therapeutics are a $ multibillion drug market. These biologic drugs are produced in non-human cells but must be shown to produce a human-like glycosylation pattern. As a result, analytical chemistry is essential to establishing lot release criteria for antibody drugs. As the first generation of antibody drugs moves off-patent, there is impetus for the development of biogenerics. Establishment of the correct N-glycosylation structure is a key challenge to demonstration of generic monoclonal antibody equivalence. In the past few years, researchers have discovered how the N-glycosylation fine structure influences monoclonal antibody efficacy3. Specifically, afucosylated antibodies have increased antibody dependent cellular cytotoxicity. Those carrying high mannose N-glycans exhibit increased clearance rates relative to those with complex N-glycans.
Numerous carbohydrate binding proteins exist, including antibodies, lectins, receptors, toxins, and microbial adhesins4. Carbohydrate binding domains present in numerous mammalian proteins dictate their interactions with glycosylated partner molecules. These protein binding interactions are mediated by carbohydrate epitopes that may be expressed on one or more glycan classes in a given biological context. Such determinants are made up of 2–10 monosaccharide residues. The blood group antigens are classic examples of glycan epitopes that are expressed in high abundances on the surfaces of red blood cells. In all, there are thousands of glycan epitopes and more to be discovered. The types of such epitopes present on the antennae of glycoconjugate glycans determine the lectin domains to which the molecule will interact. It is therefore important to keep in mind the types of epitopes that may be present when designing a glycomics or glycoproteomics workflow.
Pathogens evolve to recognize host cell surface glycans in the gastrointestinal tract and in the airways2. Hosts immune systems have evolved to neutralize pathogens by recognizing non-self glycosylation. As an example, the virulence of the influenza A virus depends on the ability of the hemagglutinin glycoprotein to recognize sialic acid containing glycans on host airway surfaces. Alteration of N-glycosylation is a mechanism whereby the influenza A virus maintains virulence in the presence of the selective pressure of the host innate and antibody mediated immune systems. Thus, the longer an influenza A virus variant exists in the human population, the greater the extent of N-glycosylation on the head domain of hemagglutinin5, 6. It is thought that the increasing degree of N-glycosylation enables the virus to evade antibody neutralization. Although the presence of a consensus amino acid sequence for N-glycosylation (NXT or NXS) is readily determined from genomic sequences, it is not possible to predict the degree of site occupancy or the glycan structures expressed. As a result, it is not possible to predict how the glycosylation influences the interactions between hemagglutinin and binding partners. Therefore, analysis of glycosylation of influenza proteins is necessary in order to understand influenza virulence.
It is essential to understand the structural information produced in each type of MS experiment7–9. To begin this process, the reader is referred to Table 1 for a glossary of glycoscience terms used in this review. Mass measurement produces information on the composition of biomolecules. In a glycomics or glycoproteomics experiment, an accurate mass may be used to calculate the general monosaccharide composition of the detected ions, i.e. Hex, HexNAc, dHex, NeuAc, etc. For this purpose, the more accurate the mass measurement, the greater the certainty of the interpretation of the glycosylation pattern. A single stage of MS is often used to profile glycoconjugate mixtures based on extracted mass and abundance information. If one makes assumptions regarding the glycan compound class (for example N-glycans), then this information may be elaborated to include the presence of structures, for example the chitobiose core, that are common to all N-glycans. It is important to recognize that such assumptions derive from information not produced directly in the MS experiment.
One may perform tandem MS on some of the ions observed in the profiling experiment. It is very important to understand, however, that such ions will likely be present as isomeric mixtures; the tandem mass spectra will reflect the mixtures present. If desired, one may purify the glycans and perform additional tandem MS experiments. It is possible in some cases to directly interpret the glycan branching and linkage directly from the tandem mass spectra. This task is complicated if more than one glycan positional isomer is present in the sample. As a result, the investigator must judge the purity of the glycan sample. Glycan tandem mass spectra may also be interpreted in reference to glycan standards of known structure. The use of such standards is necessary to validate tandem mass spectral interpretation.
The concept of sequencing is best applied to linear biopolymers. Many glycan classes, however, are branched and as such do not have a linear sequence. The analytical challenge in glycomics is therefore to determine the connectivity of monosaccharides in the glycan and the linkages for each glycosidic bond. Each glycosidic bond has an anomeric position (α or β), but this cannot be determined using tandem MS except under rare circumstances. The monosaccharide linkages present in a purified glycan can be determined using gas chromatography-mass spectrometry linkage analysis10. In the absence of sufficient quantities of purified glycan material for this method, anomeric positions are often assigned using biosynthetic assumptions. The tandem MS experiment determines Hex, HexNAc, dHex, NeuAc, etc based on mass values. The identity of the Hex and HexNAc monosaccharides may be inferred by making assumptions based on the compound class and biosynthetic rules. These assumptions are often made implicitly in publications rather than stated directly.
There are substantial differences between the two primary mass spectrometric biomolecular ionization methods with respect to analysis of glycans and glycoconjugates. Under typical vacuum source conditions, matrix assisted laser desorption/ionization (MALDI) results in dissociation of labile glycosidic bonds in glycan and glycoconjugate analytes7, 11, 12. This is of greatest concerns for acidic monosaccharide and substituents (sialic acids, uronic acids, sulfate, phosphate), and for fucose residues. As a result, use of vacuum MALDI for profiling of native glycans or glycoconjugates will typically underestimate the abundances of these labile monosaccharides in the ions detected. Dissociation resulting from the ionization process limits the usefulness of vacuum MALDI tandem MS of native and reductively aminated glycans and glycoconjugates. Permethylated glycans are considerably more stable than their native and reductively aminated counterparts, and may be profiled effectively using vacuum MALDI MS and tandem MS methods.
Because it is considerably gentler than MALDI, electrospray ionization (ESI) MS may be used to profile intact native glycans without dissociation of fragile acidic groups. To be successful in the application of ESI MS, the investigator is advised to purchase glycan standards to verify the performance of the instrument. In addition, it is typically necessary to use source desolvation settings that are gentler than those used for peptides and other analytes.
The peak capacity of MS experiments is increased by the addition of chromatographic separations, and ESI is directly compatible with on-line chromatography8, 13, 14. On-line liquid chromatography/mass spectrometry (LC/MS) improves the ability to detect low abundance glycans/glycoconjugates relative to analysis of unseparated mixtures. For this purpose, hydrophilic interaction chromatography (HILIC) and porous graphitized carbon chromatography (PGC) may be used for analysis of native and reductively aminated glycans. PGC and reversed phase (RP) chromatography may be used for reductively aminated glycans. RP, PGC and HILIC may be used for glycopeptides15. Permethylated glycans may be analyzed using RP and PGC16.
Except where indicated, the nomenclature used in this review follows that developed in 1988 by Domon and Costello17 as shown in Figure 1. Note that C- and Y-type ions correspond in mass to intact glycans with the same monosaccharide composition. The masses of cross-ring cleavages (A and X ions) help determine positions of glycosidic bond attachment, referred to as linkages. Note also that in some cases a double glycosidic bond cleavages occur on either side of a monosaccharides. These product ions, known as D ions, occur to Hex and HexNAc residues that have are substituted in the 3-position7. Analogous residues unsubstituted at the 3-position are likely to undergo A-type cross ring cleavage.
Tandem mass spectra on glycans and glycoconjugates may be interpreted directly or in reference to standard compounds. Unlike with proteomics, genomics databases are not used for the spectral interpretation. As with all tandem MS experiments, however, precursor ion isolation is a very important consideration for glycomics. Typically precursor ions are isolated with a window of 3–5 u in ESI instruments; for MALDI time-of-flight instruments, the isolation window is considerably wider. As a result, the possibility of co-isolation of background or contaminant ions must be considered. The formation of product ions from such co-isolated ions has the potential to cause false interpretation of the data. Because the likelihood of such co-isolation increases with the complexity of the sample entering the MS analyzer at any given time, chromatographic separation steps are recommended to increase confidence in the data. The use of on-line LC/MS has the advantage that the number of glycan ions entering the source at any given time is fewer than with direct MS analysis of unseparated mixtures. In addition, the abundance of each precursor and product ion may be plotted versus time. Such extracted ion abundance data are useful for demonstrating that observed product ions derive from the target precursor ion18.
Dissociation induced by collisional heating includes the collisional activated dissociation (CAD) and infrared multiphoton dissociation (IRMPD) methods. In positive ion mode, glycosidic bond dissociation occurs when a cation associates with an electron lone pair of a glycosidic oxygen atom19, 20 (see Figure 2). Protons that associate with the glycosidic oxygen weaken the glycosidic bond, giving rise to a low activation energy barrier to dissociation (Figure 2A). So low is the activation energy for protonated ions that monosaccharide residues attached to glycans and glycopeptides have been observed to rearrange during the collisional heating process7, 21, 22. As a result, it is not recommended to use such tandem mass spectra for interpretation of glycan fine structure. The extent of glycosidic bond destabilization decreases with increasing cation radius and/or affinity for sites other than the glycosidic oxygen atom (Figure 2B). Rearrangements of sodiated glycan or glycopeptide ions have not been reported. As a result, the use of sodium (or other metal) cationized ions is recommended for instances when monosaccharide rearrangements of native and reductively aminated glycans and glycopeptides must be eliminated. Typical LC/MS conditions, however, result in formation of protonated ions. Despite the fact that tandem mass spectra are generated readily using LC/MS/MS, it is risky to interpret detailed glycan structures from the tandem mass spectra or protonated precursor ions. As shown in Figure 2C, the most commonly observed cross-ring cleavages for CAD and IRMPD of glycans are those formed by facile rearrangement mechanisms. Examples include the 0,2A ion observed for the reducing end of native N-glycans. This ion is formed at the reducing end of glycans via retro-aldol condensation23. Formation of this ion is observed for 4- and 6-linked monosaccharide residues, but not those substituted in the three position. The 0,2A ions often undergo further dissociations to form 2,4A ions. Although cross-ring cleavages occur to native and reductively aminated glycans, their abundances are relatively low for cationized precursor ions. The formation of A ions is most facile for the precursor ion and following C-type glycosidic bond cleavage ions.
Glycans form negative ions either through deprotonation or anion adduction24. Glycans lacking acidic groups undergo negative polarity CAD to form a series of Cn ions, resulting from consecutive losses of monosaccharide residues from the reducing end25, 26. This is in contrast to cation mediated dissociation in which B- and Y-type ions are formed. Each Cn ion may undergo retro-aldol rearrangement in the gas phase to form a corresponding 0,2A ion, as shown in Figure 2C. The formation of such ions is blocked for 3-linked monosaccharides. For such residues a second glycosidic cleavage occurs, known as a D-type ion. These mechanisms are reliable for determination of linkages for glycans lacking acidic groups.
The situation is more complicated for acidic glycans. Such glycans ionize by dissociation of the acidic protons. For sialylated glycans, the sialic acid residues typically dissociate to produce abundant B1 ion, reducing the abundances of structurally informative Cn ions27. For sulfated glycans, the most acidic functional group is the sulfate itself. Thus, charge will tend to reside on the sulfate. A protonated sulfate group undergoes rapid and undesirable loss of SO328. Such losses are minimized for precursors in which all sulfate groups are deprotonated. In practice, it is not possible to produce such charge states for highly sulfated glycans due to repulsion between adjacent negative charges. Thus, a degree of SO3 losses is observed with tandem MS of sulfated glycans, regardless of the type of dissociation used.
When a glycosidic bond cleavage occurs for a native glycan ion, the product ion masses do not determine which bond was cleaved. Such dissociation for permethylated glycans results in the formation of product ions with unique masses by virtue of the lack of methyl or methoxy groups. Such scars, as they are known, provide high value for determination of the topology and linkages for glycans29. The permethylation chemistry30, 31 is very robust, and new variants have been published to increase sample throughput and decrease consumption32. Permethylated glycans are typically observed as sodiated ions, unless reversed phase LC/MS is used, under which conditions protonated ions are formed. Multistage tandem MS methods have been developed for gas phase dissection of permethylated glycans using ion trap instruments33–35. This approach is clearly the most powerful mass spectrometric method for glycan structural analysis. However, as with all tandem MS methods, confidence in the interpretation depends on clear understanding of the limitations of the method. There is the possibility of ion co-isolation leading to incorrect interpretations. Such concerns are reduced by chromatographic steps to reduce mixture complexity. There are also possibilities for derivatization side reactions, and separations help increase confidence in data interpretation. Also, application of high accuracy mass spectrometry helps to increase confidences in the mass assignments of permethylated glycans.
An example of the use of multistage tandem MS to determine ovalbumin N-glycan connectivity is diagrammed in Figure 3 based on published data36. The data were acquired using an ion trap mass spectrometer by selecting successive ions for tandem MS up to MS6. For such data, glycosidic bond dissociation produced abundant ions with scars, each of which corresponded to the absence of a methyl or methoxy group. The data were consistent with the presence of the two structural isomers shown. As is often the case with MSn datasets, the isomeric differences became apparent when the structure was dissected in the gas phase down to the core branching residues, the patterns of which are indicated by mass, due to the scars present. The order of losses of non-reducing end HexNAc residues cannot be determined from the data; the order shown is arbitrary. The antennae HexNAc residues were labeled as GlcNAc based on glycan biosynthetic principles and prior knowledge of ovalbumin glycans. This example illustrates the use of multistage tandem for analysis of glycans present as isomeric mixtures.
Collisional excitation of ions results from slow (on the molecular scale) vibrational heating of ions, resulting in bond dissociation. IRMPD also results from vibrational excitation of ions. Typically, the most labile bonds produce the most abundant ions using these dissociation mechanisms. For glycans, this means that the abundances of cross-ring cleavage ions that provide the information for assigning monosaccharide linkages are often low in abundance. As a result, not all linkages can be assigned in a typical CAD or IRMPD tandem mass spectrum. Although multistage tandem MS increases the information produced, the time and sample consumption scales with each additional tandem MS stage. Therefore, in order to maximize the structural information that can be produced on limiting sample quantities, it would be highly desirable to increase the structural information that may be produced on glycans in a single stage of tandem MS.
Towards these ends, researchers have been investigating use of activated electron dissociation methods. Activated electron dissociation (ExD) methods, for the purposes of this review, include electron capture dissociation (ECD)37, electron transfer dissociation (ETD)38. Each of these methods involves transfer of an electron to the precursor ion, turning it into a radical species. As has been described for peptides, such radicals undergo rearrangement to create bond cleavage. Fortunately, the types of bond cleavages observed do not depend on bond lability; rather, they depend on proximity to the site of radical production and subsequent rearrangement mechanisms. Therefore, the bond dissociation patterns observed using ExD are complementary to those from vibrational excitation (CAD and IRMPD) methods.
During ExD, a positively charged precursor ion gains an electron, decreasing the ion charge. This radical cation then undergoes rearrangement to dissociate backbone bonds. ExD, depending on the conditions, has the potential to dissociate glycans based on a mechanism that is complementary to vibrational excitation (the mechanism behind CAD and IRMPD). Studies have shown that ECD of metal cationized native glycans results in comparatively subtle differences compared to CAD tandem mass spectra on native glycans39. ECD produced A-type cross-ring cleavages that were somewhat higher in abundance than in CAD spectra. Abundances of A-type cross-ring cleavages are higher for neutral and sialylated glycans ionized in the negative mode25, 26.
The greatest potential benefit of ExD methods is to increase the dissociation yield of cross-ring cleavage ions in general and X-ions in particular. X-type ions are generally not observed using CAD or IRMPD methods unless there is a facile ene-type rearrangement, such as for unsaturated monosaccharides. The most promising results published thus far are for metal cationized permethylated glycans40, 41. Use of permethylated ions eliminates uncertainties regarding multiple bond dissociation observed with native or reductively aminated glycans. The cation has been observed to direct the product ion pattern. In particular, because the dissociation requires diminishment of positive charge, there is an advantage to the use of multivalent metal cations to maximize charge state41. The formation of a radical cation has the potential to produce both C-type ions, similar to observed using CAD, and Z-type ions. It has been observed that ETD of magnesium cationized permethylated glycans results in abundant A- and X-type cross-ring cleavages. This is consistent with the formation of Z-type ions through a mechanism that results in opening of the glycosidic ring, as shown in Figure 4. The formation of such open ring Z-type ions is significant because they lead to the formation of X-type ions. It will be of interest to see how these new results play out in biochemical applications using ExD methods.
Electron detachment dissociation (EDD) is a technique whereby an electron is removed from a negatively charged precursor ion42. The use of this technique is advantageous for carbohydrate classes that contain acidic groups, owing to their propensities to form negatively charged ions. Despite the fact that acidic glycans form abundant negative ions, the acidic groups tend to undergo facile dissociation during CAD and IRMPD. As a result, in many cases the loss of the acidic group dominates the tandem mass spectrum, and dissociation events occurring to the remainder of the glycan bonds are disfavored. Fortunately, EDD product ion patterns for such glycans complement those generated by CAD. The use of EDD improves the abundances of A-type cross-ring cleavages relative to IRMPD and CAD and produces some X-type ions43. EDD is particularly useful for sialylated glycans44 and for glycosaminoglycans45–47. The EDD method suffers, however, from poor efficiency, requiring extended acquisition times for spectral averaging. As a result, EDD has not been applied to on-line LC/tandem MS of glycans. It is possible to detach an electron from a negatively charged precursors in the gas phase using ion-ion reactions. Such negative ion ETD reactions may be accomplished in a trapped ion mass spectrometer with subsequent high resolution, high mass accuracy mass analysis. This has been demonstrated for glycosaminoglycans, for which product ion patterns are similar to those of EDD48. In summary, methods for the use of ExD for negative ions are developing rapidly and may become applicable to on-line LC/MS experiments.
Returning to the example of influenza A virus hemagglutinin, the glycosylation of amino acid residues adjacent to the sialic acid binding site strongly influence virulence and recognition by the immune system6. Thus, the understanding of influenza biology depends on the ability to determine the structures of glycans that modify specific amino acid residues. In fact, the ability to determine and exploit the biochemical mechanisms whereby glycosylation on numerous glycoproteins mediate biological function depends on the ability to determine site-specific glycan structure. Analytical methods capable of confident mapping of glycoprotein glycans to specific amino acid residues are now emerging. This section gives an overview of methods that enable understanding of how glycosylation is used as a means of modifying protein properties at specific amino acid sites.
The assignment of glycopeptide glycosylation sites, the compositions of attached glycans, and relative abundances of glycoforms is known collectively as glycopeptide mapping49. Accurate glycopeptide mapping requires a combination of techniques to provide verification of interpretation necessary to minimize false identifications50. A workflow that reflects the use of MS methods available in many proteomics facilities is shown in Figure 5. In order to produce accurate glycopeptide mapping results, it is essential to perform proteomic analysis of the polypeptides in the sample. Real world glycoprotein samples are often contaminated with other glycoproteins and/or proteases. By identifying these contaminants, the investigator will be able to generate a custom proteomics database reflecting the polypeptides detected in the sample. This custom database is then the basis for interpreting glycopeptide mass data so as to minimize false positive identifications. The value of accurate mass measurement cannot be overstated for glycopeptide profiling; the more accurate the measurement, the greater the certainty of the interpretation. Reversed phase chromatography and HILIC provide complementary LC/MS results for glycopeptides, and there is value for combining data from both methods to maximize coverage of the target glycoprotein sequence51. It is useful to conduct proteomics MS before and after deglycosylation of the glycoprotein using peptide N-glycosidase F (PNGase F). In the presence of H2180, the formerly glycosylated Asn residue is converted into 18O-Asp, and peptides containing this residue may be identified using the custom proteomics database.
The CAD and ExD methods provide complementary information on glycopeptide structure52. CAD dissociation produces abundant cleavages of the glycan with peptide bond cleavages in low abundance. The extent of glycan dissociation in ion traps is considerably less than observed in instruments with beam type dissociation cells. The latter such instruments include quadrupole time-of-flight, triple quadrupole, and Fourier Transform mass spectrometers with external dissociation cells. Dissociation of protonated glycan ions is not recommended because monosaccharide rearrangements may result. Glycopeptides, however, are ionized typically as protonated ions in MS and LC/MS workflows. As a result, it is risky to attempt to derive detailed glycan structures from tandem MS of these protonated ions. It is more appropriate to use the CAD tandem MS data to verify the composition of the glycopeptide glycan. ExD dissociation occurs primarily to the peptide backbone, leaving the glycan intact. Thus, ExD is useful for confirming the identification of the peptide. In favorable cases, the site of glycosylation may be identified.
The ability to confidently map glycoprotein glycosylation depends on the sensitivity and mass accuracy of the MS instrument. Due to the increased spectral summation times, the ability to detect low abundance glycopeptides is greater for collected LC fractions than obtainable using on-line LC/MS53. As a result, there is value in use of automated nano-ESI robots for acquiring glycopeptide mapping data.
Mucin type O-glycans are built on GalNAc residues that modify Ser/Thr amino acid side chains. This type of glycosylation occurs typically clustered into domains with many modified Ser/Thr residues. There is a substantial degree of heterogeneity of the O-glycan structure on each modified amino acid residue. As a result, analysis of mucin type O-glycopeptides poses a considerable analytical challenge54. Unlike for N-glycosylation, there is no consensus sequence for predicting the location of amino acid residues glycosylation by mucin O-glyans. In addition, there is no enzyme for universal release of O-glycans. In the face of these difficulties, there is a strong need for determining the patterns of O-glycan modification and the glycan structures at each modified amino acid residue. The most promising approach to emerge in recent years to address this problem is the application of ECD/ETD methods. Similar to N-linked glycopeptides, ECD/ETD dissociates the peptide backbone preferentially, leaving the O-glycans intact. Thus, it is possible to assign the pattern of O-glycan substitution using ECD/ETD.
A particularly impressive example of O-glycopeptide analysis is shown in Figure 655. Immunoglobulin A (IgA) molecules have a hinge region that carries a mucin-like domain with a cluster of O-glycans. In IgA nephropathy, IgA molecules are present with incomplete galacotsylation in the hinge region, meaning that some of the O-glycans are present as GalNAc with no other monosaccharides present. The research question involved determination of the pattern of expression of deficiently galactosylated O-glycans of the IgA hinge region. It was possible to determine the pattern of glycosylation for a 31 amino acid hinge region peptide that contained five sites of O-glycosylation. The ECD tandem mass spectrum shown shows a series of product ions, the m/z values of which determine the masses of the corresponding peptide fragments and attached glycans. Assuming that glycosylation occurs only at Ser and Thr residues, the m/z values of the series of product ions suffice to determine the amino acids modified by four Gal-GalNAc disaccharide groups. It was also possible to narrow the site of incomplete galactosylation (i.e. modification with a single GalNAc) to one of two Ser residues, as shown in the figure. This example demonstrates the utility of the ExD approach for determining patterns of glycosylaton in mucin-like domains. The ability to apply ExD to biological samples is improving rapidly with increases in instrumentation sensitivity and efficiency.
In summary, confident glycopeptide mapping requires acquisition of proteomics, and glycopeptide MS and tandem MS data. The interpretation of such data remains the most time consuming part of the analysis. Today there exist no software programs that integrate the diverse data types needed for glycopeptide mapping.
The structures of glycans determine the carbohydrate binding protein partners to which a glycoprotein binds. Carbohydrate binding domains are common in cell surface and secreted proteins. In order to understand fully the structure-function relationships for glycoproteins, analysis of the glycan structure is unavoidable. Mass spectrometry is an enabling technology in this area.
Accurate mass measurement determines glycan compositions; this information may be used to infer the presence of glycan epitopes based on biosynthetic principles. Although tandem mass spectrometry may be accomplished on native glycans, the quality of the product ion patterns is better for permethylated glycans. Collision based tandem MS of permethylated glycans produces primarily glycosidic bond cleavages that are useful for determining glycan connectivity. The determination of linkage positions, however, requires formation of cross-ring cleavages, the abundances of which are often limiting. Cross-ring cleavage abundances may be improved through the use of multistage tandem MS. Such analyses are not possible using accurate mass measurement at the present. The use of activated electron dissociation results in product ion patterns with more abundant cross ring cleavages than those generated using collisional excitation.
Mapping of glycoprotein glycans is best achieved using a combination of MS based approaches. It is important to acquire proteomics data to generate a custom database of the proteins in the sample. With this database, one can then analyzed MS and tandem MS data produced on glycopeptides, peptides, and released glycans, respectively. The analysis of the data remains time consuming at present. Improvements in the ability to map glycoprotein glycans are likely to come from improvement of mass spectrometer sensitivity, activated electron dissociation methods, and the availability of comprehensive software for analysis of the data.
The author is supported by NIH grants R01098050 and P41RR10888
Author biographies
Nancy Leymarie is a Senior Research Scientist working at the Center for Biomedical Mass Spectrometry (CBMS) at Boston University. Her research is focused on the development and application of advanced mass spectrometry methods in the fields of glycoproteomics and glycomics. Joseph Zaia is Professor of Biochemistry and Associate Director of the CBMS. His research is focused on structural glycosylation phenotypes related to human diseases. He has developed mass spectral methods for glycomics and is applying these to amyloid diseases and cancers. His research group focuses on biochemistry and structural analysis of glycosaminoglycans, glycoproteins, and bacterial polysaccharides.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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African American is identical to dark skin. However, their skins are not too dark. Some African American women feel difficult when they choose hairstyles. If you are an African American and you have a round face, there are many hairstyles that are appropriate for you. Most of them are with short hairs. So, if you are looking for short hairstyles for round faces African American, you can choose the following options.
Auburn Faux Hawk Tapered Sides
One of the short haircuts for round faces African American is auburn faux hawk tapered sides. Different from Mohawk, tapered will make you look sporty. With a little curl, it will look great for the crown whereas the sides are very short. Anyway, it can be considered as one of the most recommended short hairstyles for round faces African American.
Short Haircut with Side Highlighted Bangs
If you consider a host haircut with bangs, you can follow this one of the short hairstyles for round faces African American. Firstly, cut your sides hair. However, the top and front hair should be longer. After that, create left side bangs. It will look great if the bangs are highlighted. The most ideal bangs should be at about eyes length. Anyway, this chick haircut will be one of the African American short hairstyles for round faces recommended for you.
Curly Cropped Haircut with Chick Swirled Bangs
If you have curly hair, you can apply curly cropped haircut with chick swirled bangs. To apply this, you need to cut your sides hair. Then, create swirled bands covering your forehead. You can also highlight your bangs. It will make you look very chick. Those all are short hairstyles for round faces African American. If you interested, you can choose one of them that suits you.

subset: commoncrawl


text:
Tamed Teens Ely's throwing fly balls out her ass before ass fuck
- 12:15
- 352
- 2016-10-12
Ely is about to really get her holes rocked. She pops a ball out of her assole before getting assfucked and double penetrated. She swallows three big loads.
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Starting a new career - part time. Anyone done this?(13 Posts)
I'm in that situation a bit although there is possibility that something else may come up in the same company. The way I see it is a reasonably paid pt job with opportunity to add breadth to my cv.
Thanks itsall I'll look in to that.
Charities tend to be good at recruiting part time even at senior levels. They often have limited funds and would prefer to secure someone more experienced on less hours than someone full time but more junior.
We can manage on what we're earning at the moment, and we didn't want the DCs in full-time childcare so I'm not going to consider working full-time right now - but in a few years that could be an option.
Tbh it's more that I miss having a career and I miss the money. My current job is pretty dull and I don't like the thought of still being there in a year or two. However, if that's the only option then that's what I'll do.
DP works for himself so cutting his hours right now won't work.
I think, zimbah that you make a good point about volunteering - I could do something whilst on mat leave and take it from there.
I fear this may be a case of wanting everything and wanting it now!!
Do you need to earn a salary now or soon, or can you manage on your DH's salary? Some types of jobs (e.g. adult education) can be accessed either by funding yourself to do some training or by getting experience as a volunteer in that particular field and then applying for work. It really depends on whether that's financially viable for you and what kind of a career you're looking for.
I think it depends entirely on what you would want to do as a career. I used to work for a high street bank and if you were happy to start from the bottom and work your way up you could be part time from day one. However, anything you would do would ultimately be sales and customer service based so if that's not for you then it's not worth your while.
Do you have a husband/partner? Would he be willing to go part time if you can't afford full time childcare so you can concentrate on your career for a few years? If not it ultimately comes down to what matters most to you, a few of my friends with young children work full time and find it very rewarding but for me personally I wouldn't want my son in full-time childcare.
Thanks. Unfortunately i would be really bad at sales, shame as it does seem to be where the most opportunity is.
I think you are right about training positions. That's what my last job was and i was very lucky - haven't seen anything like that recently though
I think this would be very difficult if not impossible to do, especially in this economic climate.
Almost 10 years ago I secured a training contract with my ex-employer (public sector) whereby I worked for 3 days a week and had study days (either at uni or studying at home) 2 days a week over a period of 2 years. Although not strictly part time, the study days were more often than not spent at home so I did have a good work/life balance. As soon as I qualified I requested part time work and they agreed to employ me as a qualified practitioner 3 days a week.
I don't think many employers are offering training positions anymore, which imo would be the best way of getting retrained and offered a part time position but perhaps that would be something to look into?
I have since sidestepped into a different area of work and had to take a full time jobs and then request flexible working after 1 year to reduce hours.
Have lurked on many of these threads over the years and it always seems to come down to selling stuff to your mates at 'parties.'
People who have re-trained and done what you wish to do either don't come on MN (too busy) or choose not to share the secret - maybe to avoid outing.
Hope you have better luck.
Tell you one thing: if you're into fashion and have a remotely salesy personality, House of Colour have franchises available in various locations - not mine, sadly. Think a couple of MNers have started to do that recently - but I appreciate it's not for everyone.
Thanks guys!
I will stick with the current job if all else fails but wanted to take the opportunity of looking for something else while i'm not there anyway.
I would give your current job a chance - even if you don't love it, if part-time is more important to you, then you can make the most of it until your children are a little older and you can retrain and consider all options.
At 3 your eldest will have 15 hours free childcare available, which does help quite considerably. Good luck with it, I know it's tough x
I'm trying...but my situation is not retraining. More that I have no experience apart from holiday jobs. No one wants to give part time
A work experience placement (if you can cope without the money/a potential loss because of childcare)?
I was redeployed just before starting mat leave, and the job really isn't for me. The job I was in is highly specialized and in closing us down they have effectively ended my career (we are not in a position to relocate).
I want a career, as opposed to a job, IYSWIM, but I have 2 under 2 so retraining is likely to prove difficult, and even if I can - will I realistically get a decent job in a new field if I can only work part time?
Has anyone done this? Any ideas as to what I can do?
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Participate In A Program Of The CNA For Rewarding Career
December 2, 2010 by Bonjour
Filed under Certified Nursing Assistant
There are several ways to become a Certified Nursing Assistant or CNA. In a program of the CNA, there are many different courses to complete before the meeting of the CNA studies.
Different routes
A simple way to program the CNA would be a local college. In addition to vocational training school can provide the desired program CNA. If you are not available in their town or close proximity to providers of health care professional, such as hospitals or clinics for training. Some of the Red Cross also offer places to some of the training programs of the CNA, because they are short courses.
Otherwise, there are programs CNA online and you can consider, but you can check if these courses online or correspondence courses are recognized by the state or the health authorities.
Recognition by Authorities
There are many programs CNA offered to the nation itself, but different states have different requirements, although most programs CNA have some key points or key that must be completed before moving state exam CNA specific. These CNA programs nationally formulated and regulated by the supply of nursing councils in different countries and regulatory committees.
So in addition to the main features of government programs offered by the CNA to the main range. Other factors CNA program, such as cost, duration, content and materials vary.
CNA Training
Assistant in the software, there is both theory and practice. Therefore, it is important to participate in the relevant class a good understanding of the theory prior to their implementation in practice. Many programs are required to supplement their classroom training, vocational training, where you get to apply what they learn in the classroom the real working environment. Therefore, there must be some part-time messages from health care providers, such as the Red Cross.
The practical work often situations where the student would have to work with a partner or group. Works well with others to improve the capacity of assistant to care. Learning can also be video instruction to better understand what the conditions are ready for such a real impact when you,re in the field.
Program Costs
A typical program can cost up to $ 1,500 as tuition fees, there may be some financial aid or state grants or organizations of health care you may ask. Some practices you need to pay additional charges for equipment and the use of the facility. Therefore, you should check all the hidden costs or additional charges may apply program of your choice to suit your budget and requirements.
Prepare well for the exams to obtain certification that you have worked so hard. The theoretical part is to test the procedures of information and a practical assessment of the management of real-life situations.


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I have never had the opportunity to study photography in any school or institute. My background is more technical involved in the High Tech Engineering fields. I always had love for art and a great appreciation of photography. Growing up in the UK was tough, with family making ends meet. I would regularly go window-shopping for about 4 years every weekend until I got my first camera, a Nikon FM2 with a 50mm Lens, which I still own.
My Photography career really took off once I had moved to Vancouver. During my journey in photography, I have used various cameras in all shape and sizes. I was never married to a brand but used what was required for the job. I started off with Nikon, and Contax (BTW The G lenses are just outstanding) and then 2010 moved to Canon due the video capabilities. During this time I was also shooting with the Hasselblad H4D -50 and there isn't a DLSR that will match the quality.
Just due to the hype, recently I invested in to the Sony A7R and the Sony 55mm f/1.8 Zeiss lens. Within a few days I was hooked and it has become my go-to camera. Contax G glass (Contax 28mm f/2.8 Biogon, the Contax 35mm f/2, Contax 45mm f/2 Planar and Contax 90mm f/2.8 Sonnar (is just beautiful on this compact body(. Although I have a complete Canon Kit including Canon 1Dx and a Canon 5D MkIII with primes lenses, including Canon 35mm f/1.4, Canon 50mm f/1.2 and a Canon 100mm f/2.8 macro, Canon 24-70mm f/2.8 and the Canon 70-200mm f/2.8 II with various accessories but the Sony kit lives in my Bilingham 225 classic bag 100% of the time.
I just ordered the Sony A7 as a second body.
People photography has been my prime subject. I had the opportunity to travel in India for a short period capturing local tribes and my submissions were selected as a finalist in the most recent "2012 Hasselblad Masters" competition.
I have been a low-key photographer until the recent few years, exploring the social media networks to get my work out there. I have produced imagery and marketing material through film and photography for the fashion and commercial industries worldwide. My work can become very personal. I'm always looking to bend light to give that perfect look. I don't think there are bad pictures; it's just how we perceive what is good or bad. We often feel time never stops and I feel photography allows you to keep a moment from running away. People are certainly my cup tea in terms of subject and I'm always thinking "will my pictures express what is that person thinking at that very moment?"
My Philosophy:
When I stand before God at the end of my life, I would hope that I would not have a single bit of talent left, and could say, 'I used everything you gave me'.
– Erma Bombeck


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|Home | About | Journals | Submit | Contact Us | Français
OBJECTIVE--To examine the effect of physical training on physical fitness and blood pressure in children aged 9-11 years. DESIGN--Prospective randomised controlled intervention study of a sample of children drawn from a population survey of coronary risk factors in children. SETTING--Odense, Denmark. SUBJECTS--69 children with mean blood pressure greater than or equal to 95th centile (hypertensive group) and 68 with mean blood pressure less than 95th centile (normotensive group), randomly selected from a population of 1369 children. INTERVENTION--67 children were randomised to receive three extra lessons a week of an ordinary school physical education programme for eight months. MAIN OUTCOME MEASURES--Physical fitness assessed by calculation of maximum oxygen uptake and blood pressure recorded by one unblinded observer. RESULTS--After three months neither blood pressure nor physical fitness had changed significantly. After adjustment for values in weight, height, heart rate, and the variable in question before training physical fitness rose significantly at the end of eight months' training, by 3.7 mlO2/kg/min (95% confidence interval 2.2 to 5.3) in the normotensive training subgroup and by 2.1 mlO2/kg/min (0.1 to 4.2) in the hypertensive training subgroup compared with that in the controls. Systolic and diastolic blood pressures in the training subgroups fell significantly by 6.5 mm Hg (3.2 to 9.9) and 4.1 mm Hg (1.7 to 6.6) respectively in the normotensive group and by 4.9 mm Hg (0.7 to 9.2) and 3.8 mm Hg (0.9 to 6.6) respectively in the hypertensive group. CONCLUSIONS--Physical training lowers blood pressure and improves physical fitness in children and might have implications for an important non-pharmacological approach to primary prevention of essential hypertension.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Forever New
Fashion clothing and accessories retailer
Emerged late 2006 in Melbourne as a start up retailer and now operates 111 stores across 7 countries.
|Forever New Pty Ltd
Company Assessment
|Forever New Pty Ltd
This company has signed the 'Accord on Fire and Building Safety in Bangladesh', a program endorsed by Bangladeshi and international unions and labor rights organizations. The ground-breaking program includes independent safety inspections with public reports, mandatory factory building renovations, the obligation by brands and retailers to underwrite the cost of repairs, and a vital role for workers and their unions all in a legally-binding, enforceable agreement.
[Source 2014][More on Human Rights]
B grade in the Baptist World Aid Australia's 'Ethical Fashion Report 2017', which grades companies, from A to F, on the strength of their systems to mitigate against the risks of forced labour, child labour, and exploitation in their supply chains. Assessment criteria fall into four main categories: policies, knowing suppliers, auditing and supplier relationships, and worker empowerment.
[Source 2017][More on Workers Rights]
Oxfam Australia's 2016 Naughty or Nice list ranks the biggest and most popular clothing brands on what they are actually doing to protect and support the women making our clothes, especially on whether brands are bold enough to bring the list of factories that make their clothes out of hiding. This company has been dubbed 'Nice', as they have published a list of 70% of their supplier factories and signed the Bangladesh Accord.
[Source 2016][More on Workers Rights]
This company has signed the Cotton Pledge with the Responsible Sourcing Network, signifying a public commitment to not use Uzbekistani cotton. (Uzbekistan is the world's fifth largest exporter of cotton and has for decades been criticised for using the forced labour of its schoolchildren to harvest that cotton by hand under appalling conditions. This practice is organised and controlled by the central government).
[Source 2016][More on Human Rights]
This 2012 report by the Institute for Global Labour and Human Rights reveals how women sewing garments for this company at the Northern Fashion factory in Bangladesh are cheated of their maternity benefits, beaten, forced to work 14 to 16 hours a day for just 21 cents an hour, while being trapped in miserable and rat infested slums.
[Source 2012][More on Workers Rights]
This company has a number of community & environment claims on its website, mainly in the areas of charitable donations, supply chain and environmentally sustainable business practices.
[Source 2014][More on Sustainability Reporting]
This company has signed the Make Fashion Traffik Free Protocol, an initiative of Stop the Traffik Australia. Fashion companies that sign the Protocol commit to fully tracing their supply chain and to work to ensuring better working conditions.
[Source 2016][More on Multi-Stakeholder Initiatives]
Oxfam Australia's 2016 Transparency Scorecard investigates 12 of Australia's major fashion retailers and reveals which ones have published the names and locations of their factories. This company has only revealed some overseas supplier names and addresses.
[Source 2016][More on Governance]
Company Details
|Company Structure
|Private Company
Contact Details
|714-716 High St, Kew East, VIC, 3102, Australia
|03 9859 9111
Products / BrandsForever New
Forever New Womens Fashion


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Loan Document Specialist 4
- Employer
- Advent Talent Group
- Location
- Minneapolis, MN
- Posted
- Apr 15, 2017
- Jobs Outside Higher Education
- Businesses & Consultants
- Institution Type
- Outside Academe
Job Description
Position Title: Loan Document Specialist 4
Location: Minneapolis
Schedule/Hours: M- F 8 am to 5 pm - Overtime likely
Estimated Duration: 18 Months
Starts: October 14
Rate: $16/hr
The super-hero staffing team at Advent Talent is currently seeking Loan Document Specialists for a world-renowned mortgage/banking client in Minneapolis. This is an exciting opportunity for you to work with a global leader, ranked by Forbes as one of the top 10 publicly traded companies in the world, and as one of the worlds' most admired companies by Fortune magazine.
The role will be responsible for reviewing loan files for accuracy and completion. Job entails performing a variety of loan documentation duties on moderately complex loans, using comprehensive knowledge of policies and procedures for loan products. Functions include reviewing compliance for loan products; interpreting policies while analyzing applications, property and documentation; ordering all required verifications, documentation and subsequent follow ups. First shift schedule with potential required overtime. Excellent opportunity to get experience with a large and prestigious financial services organization. Pay of $16/hr. Hours are 8a-5p or 5:30p, Overtime - likely to occur and may be up to 20% Monday through Friday 7-5:30pm
Candidates with recent experience or education in accounting, business, finance, economics or mathematics are encouraged to apply. Candidates must have good written communication skills; high attention to detail and strong analytical skills. Ability to work efficiently in a production environment with very little team member interaction. Proficiency with Microsoft Word & Excel. Solid Data Entry skills (9000 Kph +). Willingness to work paid overtime as needed.
Why Advent
* Easy and convenient registration process, including options for online/video interviewing.
* Resume and job search coaching standard with registration.
* Genuinely caring and friendly staff, working together to find the right job for you.
* Competitive wages and benefits, including: employee rewards, referral bonuses, paid holidays, health insurance, weekly pay, transportation assistance, and more.
* Access to fantastic, often exclusive or unadvertised, job opportunities throughout the metro.
Advent Talent Group is an award-winning staffing and recruiting agency and a nationally certified Women's Business Enterprise (WBE). Powered by our superhero staffing team, we provide temporary, temporary to hire and direct placement services in Clerical Support, Customer Service, Mortgage/Financial, HR and Light Industrial.
Minneapolis, MN
Fri, 14 Apr 2017 14:01:58 PDT


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Central Adjective Synonyms: middle, medial, median, inner, inside
Pressure Noun Synonyms: force, compression, weight, power, strength
Pressure Adjective Synonyms: compression, pressing, squeezing, compressing, tension, stress, crushing
Business / Real Estate / Pressure Relief Valve: A valve used on hot water heating appliances to help keep the pressure in the heated tank or boiler from getting too high by allowing the relief of pressure from the valve. MORE
Technology / Rockets / Pressure Port: A threaded orifice which can be fitted with a pressure gauge or pressure transducer for determining the pressures within a motor while in operation. MORE
Science / Weather / Pressure Jump: A sudden increase in the observed atmospheric pressure or station pressure. MORE
Business / Construction / Pressure Relief Valve (PRV): A device mounted on a hot water heater or boiler which is designed to release any high steam pressure in the tank to prevent tank explosions. MORE
Science / Tides and Currents / Pressure Sensor: A pressure transducer sensing de-vice for water level measurement. A relative transducer is vented to the atmosphere and pressure readings are made relative to atmospheric pressure. An absolute transd MORE
Technology / Rockets / Pressure Transducer: Outputs electrical signal proportional to pressure. MORE


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COERR Mae Sot continued to provide and maintain technical support for the operation of community-based waste management in Mae La Camp in order to promote environmental sanitation and hygiene among refugee communities for their own benefit.
Waste management in Mae La includes garbage collection, sorting, reuse or recycle, burial or incineration.
Waste management covers collection, sorting, disposal and reusing of waste. Meetings are regularly held to bring cooperation and coordination among organizations, community leaders, waste management working group, school teachers and religious institutions, etc., concerned in waste management.
The environmentally-friendly households were visited and guided about waste minimizing through reducing, reusing and recycling procudure.
On February 18-19, COERR staff and Karen Environmental Group together with volunteers cleaned and cleared the waste management area and put all tools in place.
Later, staff and refugee volunteers cleared the water treatment pond by taking away Water Hyacinth which are floating and blocking waterway. These plants will be used for composting and distributed to farmers. Then the area around the pond were clear and clean.
Clean-up Day was set on March 13, in which refugees numbering 150 from Zone A, 200 from Zone B and 200 from Zone C contribute to the garbage collection in the stream throughout the Mae La camp. NGOs including COERR support truck for garbage collection, garbage bags, food and drinking water, etc.
Waste management is handled by the cooperation among many parties concerned: Government and private sectors, refugee community, Thai village, schools and religious institution, to minimize problems and impacts incurred as well as to initiate the potential of recycle waste bank project


text:
Listen to free music played by ccthefox
Blip.fm is internet radio made social. It's easy to search for, play, and discover free music recommended by real people. Join today to create your own free station and start listening to @ccthefox.
Crystal Stilts – Flying Into the Sun| play
Jun 19, 2013 at 4:55 pm via YouTube | AMAZON | ITUNES
What other DJs are saying about Crystal Stilts - Flying Into the Sun
ed_die_tryin crystal stilts – flying into the sun
Apr 5, 2017 at 1:47 am via YouTube | AMAZON | ITUNES


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Evolved with years of experience, this is a basic yet practical and a very popular design suitable for many bedroom sizes.
This is a comprehensive and practical design suitable for large bedrooms. With lots of storage spaces and compartments, this is ideal for the master bedroom.
This is another comprehensive design suitable for a Walk in Robe. With so much storage space and compartments, this is ideal to store everything in style in the master bedroom.
To organize your clothes and beautify your bedroom at the same time takes a Magic Touch. Fully customized to make maximum advantage of available space, Magic Touch wardrobe doors feature slim, softly rounded aluminium frames in the latest designer colors and finishes.
Round modern appearance with safety, design flexibility and a 10 year guarantee.


text:
- Treatments
- Breast Implant Revision
- Pictures
- 25-34 year old woman...
25-34 year old woman treated with Breast Implant Revision
*Treatment results may vary
Was this photo helpful? {{ voteCount + ' other' + (voteCountOthers == 1 ? '' : 's') }} found this helpful
This 45-year-old woman underwent a breast revision and lift around the areola to increase her breast size and correct a significant animation deformity. A 605 cc high profile smooth round implant replaced a 350 cc saline implant and a split muscle technique was used to nearly eliminate the animation deformity.


text:
Among all the peoples of the ancient world, the one whose culture most clearly exemplified the spirit of Western man was the Hellenic or Greek. No other of these nations had so strong a devotion to liberty or so firm a belief in the nobility of human achievement. The Greeks glorified man as the most important creature in the universe and refused to submit to the dictation of priests or despots or even to humble themselves before their gods. Their attitude was essentially secular and rationalistic; they exalted the spirit of free inquiry and made knowledge supreme over faith. It was largely for these reasons that they advanced their culture to the highest stage which the ancient world was destined to reach.
- Edward McNall Burns, Western Civilizations: Their History & Their Culture, pg. 147


text:
Hi. We moved systems, and because it was impossible to achieve feature parity between the systems, we opted to archive the old B&S threads but allow users to read them so that they can put them into our new B&S system.
There is a support thread if you have any issues.
Suede Leather NATO Watch Straps in 18mm, 20mm, 22mm & 24mm- $29.95 Shipped!
Discussion in '2011-2017 Streetwear' started by dicee, Jan 14, 2013.
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- Not open for further replies.
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text:
sailing ship concept
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Sculpted in 3d-coat, rendered in Keyshot.
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Salarpuria Sattva Pa...
₹ 3,500,000.00
₹ 3,543,000.00
Mahagun Group is presenting new residential project Mahagun Mywoods at Greater Noida West. It is proposing 2/3/4 BHK luxurious apartments with large area from 935 Sq. ft. to 1810 Sq. ft. within economical price with amenities and facilities.
Basic Amenities:-
· Club House and Gym
· Kids play area
· Swimming pool
· 24 hours power backup
· Internet/Wi-Fi connectivity
Advantage of Location:-
· Close to schools and hospital
· Near to Sector 18, Atta market
· Easily accessible proposed metro station
More Info:-
Email id – [email protected]
Contact No – 9582810000
Location – Sector Omicron 3 Greater Noida
Apartments Type – 2/3/4 BHK
Published date:
12/10/2014 6:08 pm
Expiration date:
Uttar Pradesh
|For sale
|New construction
|No. of bedrooms
|No. of bathrooms
|Square Meters
Safety Tips
Region: Uttar Pradesh
City: Noida
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Salarpuria Sattva Pa...
₹ 3,500,000.00
Experion Windchants ...
₹ 22,525,000.00
Adani Samsara Floors...
₹ 15,641,600.00
Best Luxury Apartmen...
₹ 4,381,000.00


text:
Download app Auto Memory Cleaner Premium v1.8.2 Full Apk latest version is here
Auto Memory Cleaner is application which cleans your memory effectively to boost speed and performance of your device.
Your running processes and their tasks are restarted and not killed what is much more effective and friendly to your device than any other task killer or memory cleaner. This method prevents the memory overflow and also boost speed of your device. Auto Memory Cleaner is running in background and does all the work for you. You can set if you want to clean memory in specific time or everytime when screen is turned on/off. Auto Memory Cleaner also allows you to restart any running process manually.
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- Auto memory cleaning when screen goes ON – Premium version
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Auto Memory Cleaner Premium Screenshot:
What's new in this version:
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Requires Android 3.0 and up
More info and screenshot from Google Play
Download link Auto Memory Cleaner Premium v1.8.2 APK:
Zippyshare | Mirror link
Topic: apps tools apps utility auto memory cleaner premium apk


text:
Talking Deadpool
Paul, Wayne, Nick and Robert gather to talk about Deadpool, the movie, the X-men universe and connections, the comics and our hopes for the future of the Deadpool and X-men franchise. We look ahead to Civil War, Batman V. Superman, Daredevil, and Star Wars Episode VIII and Rogue One.
Theme music: http://youtu.be/ALUvrEfzzgM
Podcast: Play in new window | Download


text:
A hoverboard is a two-wheeled self-balancing scooter which has actually ended up being very popular in the past couple of years. It is also considered as one of one of the most preferred presents throughout the holiday and also is similarly popular amongst children and also grownups. It is prominent since it is convenient to make use of and also has no hand-operated movements like skateboards.
Additionally known as hoverboard for sale or segway, these self balancing boards utilize sensing units to detect the instructions in which the motorcyclist leans, then its electrical motor rotates the wheels to relocate that instructions. All one should do is change his weight in the instructions he intends to relocate. You could utilize this board to commute to function or just for enjoyable to head to any type of place you want. If you are willing to acquire one either on your own or as a present for someone else, here are a few things you need to consider.
– Price- Do not go for hoverbaords which look too affordable to be true. Inexpensive ones might have a faulty battery and also you may obtain harmed while riding on them. Do proper research study prior to acquiring any product.
– Wheel Dimension- The wheel dimension ranges from 4.5 to 10 inches. Tiny wheels are advised for people that intend to ride on smooth and leveled roads. Larger wheels are good for riding on the top of soil or yard.
– Attributes- You can currently discover hoverboards furnished with features like, Bluetooth audio speakers, GENERAL PRACTITIONER as well as push-button control. All these features normally add to the cost of the item, however if you desire them, spending some extra money is worth it.
– Where to Buy- Where to get a top quality hoverboard is an essential factor to consider as buying it from an unauthorized dealership is not secure and can bring about mishaps. Acquiring a genuine hoverboard from a reputable supplier could assist you have a risk-free as well as comfy riding experience. You can additionally look for top quality hoverboards online.
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Though hoverboards are a lot of enjoyable to utilize, they can likewise be high-risk if the best safety measures are not taken prior to acquiring them. Consider the above mentioned indicate make a safe and also enlightened purchase.
You could seek floatingboard Hoverboards on-line sale to find high quality equipments. Visit our website to recognize even more about where to buy a top quality hoverboard.


text:
SAHABA 'The Blessed'
At the beginning of the book (The Blessed) superiority of Ashâb of our Prophet, Muhammad ´alayhissalâm, is explained along with how unjust and ignorant are those who defame Ashâb-ı-kirâm. Besides, the meaning of ijtihâd is explained. In the part of cautioning, an answer is given to the book (Hüsniyye) written by an enemy of Islam. In another part, biographies of great savants of Islam - hadrat Imâm-ı Rabbâni and hadrat Sayyed Abdülhakîm-ı Arvâsi - are explained. In the part Two Apples of the Eye of Muslims superiority of hadrat Abû Bakr and hadrat Umar is explained; in the part The First Fitna in Islam events between Ashâb-ı-kirâm are explained beautifully from the pen of hadrat Imâm-ı Rabbâni Ahmad Fârûkî Sarhandi who explains that to love all of Ashâb-ı-kirâm is a fundamental condition of being Ahl-i-sunnat.
This book contains 384 pages. You may request the book from the bookstore Hakikat. In order to read the full version of the book click on the picture or title.
- Login to post comments


text:
Meet the Speaker: Shefali Patel
Session: GE's Digital Transformation
GE's Digital Transformation
Digital transformation and disruption are forces that most business leaders contend with today. They traverse a vast landscape of fundamental business changes an organization undergoes to evolve, and incumbent practices that signal the need to change in the first place.
In this talk we will address multiple tenets of a digital imperative from GE's point of view. Becoming a digital industrial company has been a cultural and strategic shift that required investing in a new growth engine outside of GE's core business.
But what prompts an organization to fundamentally embrace a new business model? How does a 124 year-old company uphold a commitment to behave and think like a contemporary start-up? Answers lie in the leadership mind-set and conventional practice of market and new technology pulsing that have sown seeds of reinvention through business and technology strategy for years.
In 2011, we identified big data visualization and insights, and the digital platform-driven business model as sources of creating new value for GE and our customers, which has unleashed a new world of industrial internet. Today, we are scaling a digital industrial company by investing in technical and commercial capabilities, attracting new talent, and maintaining a relentless focus on innovation, speed and measurement. This endeavor is a co-creation effort with our customers across different sectors, who have embraced digital mind-set and are piloting our digital solutions to drive productivity.
It has been a tremendous journey that has just started. We look forward to sharing its arc and lessons learned, and benefit from your perspective on your digital transformation.


text:
Listen to free music played by mastakate
Blip.fm is internet radio made social. It's easy to search for, play, and discover free music recommended by real people. Join today to create your own free station and start listening to @mastakate.
What other DJs are saying about Remy Zero - Fair
mllomlscwltapd you always said they end up together. i call bs.
Nov 4, 2014 at 2:44 am via YouTube | AMAZON | ITUNES


text:
The IC-E90 is a tri-band FM handie designed to operate in the 6m, 2m and 70cm amateur bands. For some while there has been interest in a modification which would allow these sets to work in the 4m band too.
A modification published on-line a couple of years ago merely described how to remove the out-of-band inhibit function, so allowing the set to transmit and receive at 70MHz. This took no account of spurious outputs from the transmitter, and one correspondent had interference problems reported by local military air traffic control!
Happily, one of the major UK dealers can now supply these sets ready-modified for 4m use with the inclusion of a filter to reduce spurious outputs, although they can only achieve a second-harmonic suppression of around 33dB, so it is not advisable to use the rig into a external antenna or amplifer without additional external filtering.
Steve G1KXP recently acquired one of these sets, and got together with Ian G6TGO and Dave M3CRV to give it a try on 4m. Here is their test report:
These tests were conducted from the QTH of G6TGO near Sale in Cheshire, mostly using a Sirio vertical antenna at 15ft asl. The E90 has an SMA antenna socket, so a patch-lead was used to convert this to an in-line BNC socket (inter-series adapters are available, but there is always a concern that when attached to a heavy cable, they put too much stress on the small socket on the top of the set).
The receive performance on 4m was very good: off-air signals were a little stronger than on an Ascom SE550, and no breakthrough was heard from local Fire Brigade services in the adjacent band. (Over the course of extended tests in the following weeks, some breakthrough has been heard at the top end of the band from a nearby Fire Service transmitter on 70.525MHz, which worsened as the battery voltage dropped below 7V.)
Some chopping of signals on 70.475 was heard, but this was attributed to an amateur packet link on 70.4875 from a station only half a mile away.
The squelch is adjusted by pressing a function button below the PTT and turning the knob on top of the unit. The settings available are: OPEN, AUTO,or pre-sets 1-9.
Initial tests at Steve's QTH had indicated slight distortion on the transmitted audio, due to over-deviation. It was then found that the set has an internal menu to select narrow ("off") or wider ("on") deviation. After this adjustment, the audio reports were very favourable, with listeners commenting that they would not have known it was a handheld with an internal microphone.
The maximum RF output power available when using the supplied battery was 2.75 watts (measured on a AV600 bridge), dropping to 2.5 watts on "low" power (on the other bands, there is a much greater differential between the "high" and "low" settings). This was sufficient to have clear contacts with several stations over 20km distant.
The actual contacs made on 4m, using 2.75W into the Sirio were as follows:
Dave M3CRV in Northwich (23km): S5 with QSB.
Roger M0DWQ in Warrington (20km): RS 58.
Phil G8HDS in Rochdale (24km): RS 57, (and 41 with QSB on the set-top helical)
Paul G7ODJ in Warrington (20km): RS 58
Ross G6GVI in Bolton (20km): RS 54
During a brief test on 2m, some breakthrough from strong local paging transmitters was heard when using a dual-band Watson-50 colinear, but this was not as severe as on a handheld scanner coupled to the same antenna.
The portable antennas supplied consisted of a single-band helical for 4m, and a second three-part helical (with the longer top-section it is suitable for 6m/2m/70cm, and with the more compact shorter top it functions as a dual-bander for 2m/70cm).
The 4m helical looks a little out of proportion to the E90 but appears to function well: several contacts have been made over 40km (and one over 70km) using it.
The 4m helical is supplied with a small rubber washer which should be fitted to fill the gap between helical and the SMA connector on the top of the set (be careful to look in the packaging for this small washer when first unpacking your set).
As with all compact set-top aerials (which are small compared with the wavelength), the efficiency is poor, but can be improved by adding a counterpoise to the set. Holding the handie on top of a metal structure such as a car-roof or handrail can help, or why not try fixing a quarter-wavelength (around 40 inches) of wire to the rear belt-clip screw? You may find that as well as improving the effective radiated power, the improved antenna matching will reduce the current drawn, and extend the battery life too.
The set's receive coverage is from HF through to 999.999MHz with wide-FM, narrowband FM and AM modes. Although the set will not receive SSB or CW, when switched to AM, and with a local signal generator used as a BFO, it was able to detect the GB3BUX FSK beacons from Buxton (30km) on both 70.0 and 50.0MHz.
The radio provides 500 memory channels, plus up to five calling channels and 50 scan-limit memories. The UHF TV Band audio frequencies (channels 21-69) are also pre-programmed into memories.
As well as operating as a self-contained handie with integral batteries, the set may also be operated from an external DC supply between 5.5 and 11V (the manual warns that damage will result if this limit is exceeded). A DC-DC converter will therefore be needed if you want to operate from a 12V battery supply. The transmitter will draw up to 2A at full output, and 800mA at 0.5W output.
The transceiver incorporates a battery voltage indicator: with the supplied Li-ion BP217 (7.4V, 1300mAhr), this showed 8.6V when fully charged, and 6.3V when discharged, just before the set shut down. The supplied plug-in charger will re-charge this battery in 16 hours, but cannot supply enough current to power the transmitter. A fast drop-in charger is available as an accessory: this will re-charge the battery in 2.5 hours.
The receiver draws up to 220mA at full audio and 100mA when squelched, dropping to 65mA with the battery saver enabled.
Another accessory is available: a battery case which includes a DC converter to generate 5.5V from just two AA cells. This is sufficient to allow the set to operate on receive for a few hours, and is useful if you do not want to expire your main battery whilst monitoring. It will also manage brief transmissions at a hundred milliwatts output.
The IC-E90 is the only dedicated amateur portable FM transceiver currently available with 4m capability (on UK import models).
The combination of four bands in a small lightweight unit will make it a popular choice with those who like to get out and about with their radio, particularly SOTA enthusiasts.
Its performance is limited by the set-top helical aerial, but if connected to a more effective external antenna, its receiver performance is at least as good as most of the (ex-PMR) sets in widespread use on the band. On transmit, the 2 to 5W output is sufficient for making contacts over tens of kilometres (or over a thousand, if you're lucky with the sporadic-E!).
Note that this review has concentrated on the set's performance on the 4m band only - the rest of its capabilities have not been fully assessed.
Steve purchased his set from Martin Lynch & Sons, and it was supplied with the helical antennas shown above. Having seen and heard it in action, Ian and Dave were straight on the 'phone to order theirs!
The tests were performed by Ian G6TGO, Steve G1KXP and Dave M3CRV, and the review was compiled with additional input from Ross G6GVI.
Thanks also to the other stations who helped with on-air reports, and also to Chris and the team at Martin Lynch & Sons for their technical assistance.


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Charleston, VT466 Center School Road
Est. Mortgage: $/month
MLS Number 4659687
- Residential Single Family
- Bedrooms 2
- Bathroom 2
- Stories 1
- Construction Existing
Private home on 10 acres of open fields with over 1800' road frontage and bordering 4,000' along the Clyde River. This home offers 2 bedrooms and 2 baths, office, and open living area leading to a large (668 sq ft) wraparound porch overlooking the river and a private pond. Full walkout basement with radiant heat has been used as a workshop. Oversize 2+ car garage with a workshop above, large (32x80) pole barn for storage or animals, separate studio (which could be used as a bunkhouse for the kids), and storage shed. Trails through the woods along the river and marketable timber on the 75+ acres of woods. Great wildlife area, direct VAST trail access. New steel roofing materials for house and garage included. Broker-Owned.
Listed by Daniel Maclure and Nicholas Maclure of Century 21 Farm & Forest
Virtual TourSave FavoriteSchedule A VisitAsk A QuestionMore Details
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Est. Mortgage: $/month
Federal 30-year interest rate: 4.03% last updated on 2017-07-13
This product uses the FRED® API but is not endorsed or certified by the Federal Reserve Bank of St. Louis.


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From secret histories to off-the-beaten path landmarks, WETA Neighborhoods reveals the lesser-known aspects of our favorite greater Washington surroundings.Watch preview →
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Ten minutes into this Sunday's Skypecast, I knew I desperately needed some help. Sure, Marian from Germany helped to handle the virtual microphone, and Paulino, Argentine leaving in the U.S. offered his hands to handle the text message board. That freed me to focus on hosting. Still, I was in deep water.
Over 30 something people joined us at the beginning, and it grew to about 50 in the peak. I made a snap decision not to ask all to introduce themselves one by one to the group, as I did before, and not to ask some of them to share a news item in turn with the listeners. Otherwise, we could have spent whole two hours just to read off a long roster. So I appointed myself to be the king, choosing a few news items that I thought would be interested to most listeners.
Here are the topics I shared with the group: Sony's latest game console Playstation was selling through the roof worldwide; O.J.Simpson's latest supposedly confession book was canceled by the publisher because of the outcries from the public; the daughter of the U.S. president Bush was robbed in Argentina; and a mother nursing a baby without a cover was ordered to leave off the airplane she boarded.
I swear, if anyone had a crystal ball for choosing right topics for a dynamic group discussion, I'd pay a fortune to get it. Because no matter how encouraging I was to urge the listeners to share their views, almost no one, out of 50 some people, cared to speak up.
Then came a Brazilian. The moment he opened mouth, I knew exactly how, in a fable, the naked emperor felt when a boy called out "but he has no clothes on." After hearing me rambling on the Sony's Playstation, the Brazilian said in frustration: "Hey, you. Talk something funny; we Brazilians don't like serious topics."
I congratulated myself silently for not bring up the first two news items originally on my list: Nobel peace price and microfinance helping the poor, and the latest political assignation in Lebanon.
Fortunately, a few found it's funny that the petty thief in Argentina could get around the bodyguard and snatched the purse out of Bush's daughter. It got the conversation going for a while. Then it dried up. Even the news of a young mother's milk to nurse a baby in public wouldn't get the conversation flowing.
Marian and Paulino stepped in to help. They thought the funny movie Borat would lighten up the group. So they put out a YouTube link that shows the trailer of the movie, and asked everyone to share his or her views about the movie. I didn't know whether the trick worked or not, because I stepped away for a few minutes and, when I came back, everyone was talking about how to make a billion dollars in one year, like the founders of YouTube did.
Making a billion dollar in a year? That's funny to me.
(Edited and links by Paulino)
27 Nov 2006
Home » Sunday Skypecast » Who Has The Crystal Ball?


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How Can Food Babe Sell Carrageenan So Cheaply? It's Simple: Volume!
The calendar on the wall says it's Friday! That's payday here at Bad Science Debunked headquarters, which means it's time for a crowd favorite: another group shopping trip to FoodBabe.com. Vani Hari, the "Food Babe", earns a sales commission from purchases we … Continue reading


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Events, Ceremonies & Assembly Halls
Brussels ExpoBuilt at the occasion of the World Expo of 1958, Brussels Expo is an exposition site with 12 halls, located at the Heyzel in the center of Brussels. In 2008, the management of Brussels Expo decided that the Brussels Expo needed a new paging, evacuation and BGM system with voice alarm products and approached Prévost System Integration(PSI), TOA's exclusive distributor in Belgium and Luxembourg. Their requirements are: 1) A more flexible and failsafe system and 2) A graphic interface providing easy-to-use controller software, making all actions and hall reservations easy to put into the system without impacting on security issues and logs. Then PSI proposed the SX-2000 Network-enabled Voice Evacuation System complied with EN 60849 and EN 54 standards. First of all, PSI invited the management of Brussels Expo to visit some of reference projects in France and they were convinced that TOA could provide them with a system that satisfied their demands. Secondly, a custom-made user software was developed. The user can control and change easily the system. Finally, 6 pieces of SX-2100AI, 6 pieces of SX-2100AO and VP Series power amplifiers were installed in 3 racks. 26 Remote Microphones are placed around the Expo's various halls and patios areas. The Brussels Expo now has a state-of-the-art new sound system.
The Sultan Salahuddin Abdul Aziz Shah MosqueThis is known as the Blue Mosque, due to the beautiful blue building, which is the largest mosque in Malaysia. At the mosque, an active line array speaker delivers high-quality sound intelligibility.
Cathedral Basilica of the Assumption of the Blessed Virgin MaryThis is the cathedral located in Pelplin, a town in northern Poland. Given the very large space and complicated architecture of the building including sacred edifice, the sound design was a challenge. However, TOA's sound system achieves premier sound quality and intelligibility.
KOBE LuminarieThe beautifully lit Kobe Luminarie has been held every year to symbolize the ongoing restoration and reconstruction of Kobe following the devastation of 1995's Great Hanshin Earthquake. Adding a poetic charm to winter in Kobe, it now attracts millions of people from all over Japan. TOA has taken responsibility for the Luminarie's main sound presentation from the very first year, and has made the lights seem brighter and more colorful by creating a musical accompaniment specially suited to each year's new theme. TOA's advanced technical know-how helps create a visionary world, with the music changing as people stroll through the arches of light, and sound effects scattered across the entire site, all culminating in an inspiring harmony that brings the closing ceremony to a poignant end.
Istiqlal MosqueIstiqlal Mosque, located in Jakarta, the capital of Indonesia, the world's largest Islamic country, is the biggest mosque in Southeast Asia and can accommodate 16,000 worshippers. TOA installed a public address system in this giant mosque that features column speakers.
Expo Aichi, Earth Tower Nagoya CityPart of Aichi Expo (Japan's first twenty-first century exposition), the Earth Pavilion of Nagoya City was equipped with TOA emergency broadcast and CCTV systems that helped ensure visitor safety and security.
Sagrada Família BasilicaConstruction on Barcelona's Sagrada Família basilica began in the late 19th century under the direction of architectural genius Antoni Gaudí, and while still under construction even today, it has been named as a world heritage site. Inside this 170-meter high, reverberation-prone space, TOA Line Array Speakers designed to match the sacred interior offer clean, clear sound ideal for delivering daily sermons and choir music to visiting worshippers.
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Hyderabad: BJP has launched a campaign to propagate against Triple Talaq issue. It has started organizingdoeswareness programs with the help of the organizations supporting its ideology so that the stand taken by Muslim clerics could be ridiculed.
Pragna Bharati organized a symposium at Telugu University, adjacent to public garden, Nampally yesterday on "Triple Talaq: Justice and Social Change".
Justice CV Ramulu, Ms. Shazia Ilmi, Mr. N. Ramachandra Rao addressed the symposium. The speakers heavily criticized the stand taken by All India Muslim Personal Law Board and claimed that the decision of the court would depict social change.
Justice Ramulu termed Triple Talaq as cruelty and said that it is wrong to associate it with religion. It should be seen not from the point of view of religion but from legal point.
Ms. Shazia Ilmi told that the stand taken by the Muslim Ulema is against the Islamic Principles. She also said that Triple Talaq is a sin and punishment should be prescribed for it. She alleged that the Muslim clerics are misinterpreting the meaning of the Quran. She also claimed that the Indian Muslim women got enlightened after the Shah Bano case. She defended the stand taken by the stand taken by Govt. of India.
Mr. N. Ramachandra Reddy, BJP MLC told that any attempt to stop atrocities on Muslim women does not come under the intervention in religion. He requested the lawyers to discourage divorce cases and work for compromising the couples.
–Siasat News


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Western Nevada College career counseling
Think about what career is best suited for you, and where you want to study. Request more information now.
Think about what career is best suited for you, and where you want to study. Request more information now.
Think about what career is best suited for you, and where you want to study. Request more information now.
Get the facts about Western Nevada College career counseling, and registering for elective lectures. Which college degrees give you the best chances of finding employment after graduation? Browse accredited business management programs online. You can train in as little as two years to begin your career as a financial professional. Concentrating on Western Nevada College career counseling, for example, you may gain an advantage in corporate recruiting being done on-campus. Check with the workforce development department for course registration deadlines, and career training program requirements. Please use the form above to request admissions info for Western Nevada College.
As the economy improves, many firms are beginning to hire. Companies are in need of candidates that can fill a particular job position, so set up an introductory meeting to ask questions about the firm's history, and current business opportunities. Review your prior work experiences, internships, volunteer experience, interests and hobbies. For each item on your list, identify the skills and knowledge you developed, because many skills are transferable to the job you're applying for.
Before a first interview, analyze the open position that you are applying for. Review the job description carefully, and highlight both the employment qualifications and job responsibilities. At the same time, learn as much as you can about the company's mission, corporate services, product lists, and main competitors. Start your research on the company's website, then do an article search on Google, and attend career information sessions and job fairs that are held on campus. Beyond major requirements within the workforce development department, you may consider Wayne County Community College online classes 2017 as a complementary elective to your primary course of study.
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Torchy's Tacos Ranked Among 50 Best Tacos in America
No matter if it's from a truck, a gas station, or served in a gourmet restaurant, everyone has a singular vision for the perfect taco. Just in time for Cinco de Mayo, The Daily Meal compiled a list of favorites, deeming them The 50 Best Tacos in America. Hot damn, that's a mighty claim!
The Trailer Park taco from our own Torchy's Tacos was ranked #4 among taco offerings nationwide. We are both proud and pumped to be among such taco greatness, including several top-notch establishments from our own home state.
(For the purposes of the article, the authors define a taco as "meat or vegetables placed onto a warmed tortilla, with the express purpose of folding it and eating it with one hand." Sorry, Choco Taco, but you're out. They also excluded mobile taco trucks, probably due to sheer volume.)
From The Daily Meal:
#4 – Torchy's Tacos, Various Locations: Trailer Park Taco
At Torchy's, which has locations throughout Texas (centered in Austin, Dallas, and Houston), owner Michael Rypka has created tortilla-bound concoctions that are nothing short of devilish, and made fresh every day. There's the Brush Fire (Jamaican jerk chicken, grilled jalapeños, and mango), the Dirty Sanchez (scrambled eggs with fried poblano chile, guacamole, escabeche carrots, and shredded cheese) and The Republican (grilled jalapeño sausage with shredded cheese and pico de gallo), but the wildest and most delicious creation on the menu is the Trailer Park, a massive, battered and fried hunk of chicken breast, topped with sautéed green chilies, lettuce, pico de gallo, and shredded mixed cheese on a flour tortilla from El Milagro, topped with poblano sauce. If you prefer to "get it trashy," they'll replace the lettuce with cheese sauce. And who needs lettuce when cheese sauce is an option?
Check out the full list HERE.