text:
On Saturday mornings, Patrick Andriamahenina takes a group of 14 excited Malagasy kids through their basketball paces. But these are no ordinary lessons: Most have never touched a basketball before, and Andriamahenina is not using the lesson to teach them sport; he's trying to teach them to code.
"We're drawing out the similarities between basketball and coding," Andriamahenina, a local Malagasy basketball coach, told me in French over the phone. "For example, while playing Scratch (a free programming language), kids have to think strategically and make quick decisions. It's pretty much the same thing in basketball."
Dream Camp—where 42 kids are currently receiving lessons—is a project geared to teach underprivileged Malagasy kids coding, conservation, sport, and hygiene. It was set up in November 2015 by Chris Corbett, the founder of Human Development League (HDL), a non-profit organization based in Madagascar. The Malagasy are an ethnic group that forms almost the entire population in Madagascar.
The camp provides training to kids aged between seven and 12. To set up the pilot operations, Corbett teamed up with Andriamahenina, Malagasy coding guru Sahaza Marline, and basketball coach Cray Bony, who has run basketball camps in both the USA and Tanzania. The training venue is at the local technical high school, which has a basketball court but only five very old computers for roughly 1,000 students.
"We really wanted to give these kids some coding skills that could help with future employment," Corbett told me. In a report, UNICEF states that out of the 100 children that attend primary school only 60 percent graduate, and only 25 percent go on to complete junior high school.
When kids play with
they create animations, stories, and characters by snapping blocks together. Corbett and his team took this methodology and moved it onto the basketball court, getting the kids to give instructions to one another, envision they were blocks, and move along the court accordingly. The idea is to build a symbiotic learning relationship between the games that they conduct in the real and virtual worlds.
Corbett developed his ideas of using sport as an educative platform by attending various events across Africa that promoted development via sporting activities.
"Everyone believes that sport has inherent benefits as it promotes team work, but there is this other model which basically uses sport-based activities as metaphors to teach concepts, so that it's more than just sport," he said.
As Corbett planned to upscale his sports development projects by finding funders and partners, Madagascar was plunged into a political crisis in 2009, which resulted in street protests and dozens of deaths in Antananarivo, the city's capital. Following the 2009 political coup, economic sanctions were placed on Madagascar, bringing an already feeble economy to its knees.
Not to be defeated, Corbett continued to make connections with the local sporting, conservation, and tech community in Madagascar, until he had enough manpower to start the Dream Camp pilot project last November 2015. His first batch of students have just completed their first round of courses, and Corbett hopes to bring in more students and provide more advanced courses to his existing students.
Madagascar is currently home to a nascent tech community, boasting a modest innovation hub dubbed Habaka and a CoderDojo (a global movement of free coding clubs for kids) in Toamasina, a town on the east coast of Madagascar. According to Corbett, though there are companies in Madagascar looking for employees, access to investment and talent is still scarce. He launched Dream Camp out of his own pocket and is currently looking for funding in order to sustain and expand it.
While Corbett and his team are making up the Dream Camp curriculum as they go on with feedback from the kids, they have high hopes for what it could be in the future with the help of more partners and equipment. At the moment, the camp is operating with seven donated laptops.
"Our hope is that we can maintain these kids in our programme for multiple years, and that we can maybe link them up to our partners and companies so that they get get internships, or more advanced training," said Corbett. "We'd like to rebrand Madagascar as Silicon Island and bring in contacts from Silicon Valley and give Madagascar more visibility."


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There was an upset in Hollywood this March when the Academy anointed Sandra Bullock best actress for her role in "The Blind Side." The controversy hinged on the fact that she, an historic B-level comedienne, bested Meryl Streep, a far more Serious Actor who has a standing relationship with Oscar.
But after seeing "The Blind Side," it's clear that any outrage would have been more appropriately directed at the insidious racist and classist narrative that Bullock's character delivers, albeit cloaked in fitted, matching coordinates of Southern charm and irresistible pluck.
The film is about a young African American man who is propelled from his humble roots to NFL stardom by a wealthy white family. It's based on the true story of Michael Oher, a lineman for the Baltimore Ravens, who, as a teenager was invited to attend Briarcrest Christian School — an institution that was both physically and culturally far away from his impoverished Memphis neighborhood. The invitation came from benevolent, white, middle-class members of the faculty and staff who, after taking one look at Michael's imposing frame, saw dollar signs and championship titles in their eyes.
While struggling to fit in at Briarcrest, Michael meets the Tuohys — a kindly white family that has built its fortunes by running a string of Taco Bell franchises. They love Jesus a lot, but they love college football even more. They encourage Michael to play and later, after he's made himself a star on the high school circuit, they encourage him to play for their alma mater, Ole Miss, where they happen to be generous boosters.
Yes, these events did in fact unfold in the real world: The "privilege knows best" moral implicit in Michael Oher's story was indeed imposed upon him, and a group of Memphis elites actually manipulated his circumstances to further their interests. However, that's doesn't give Hollywood license to portray his life through a series of overwrought and oversimplified vignettes that make Bullock's character (the Tuohy matriarch, Leigh Anne) look saintly and heroic — and really fabulous while she's doing it. She even teaches him how to play football! Meanwhile, Michael is portrayed as a black kid who managed to "get away" from a drug-addicted mother, the dangerous neighborhood and the thug life because he's gentle and sweet, and because some wealthy white folks took mercy on his soul.
Unfortunately, audiences aren't meant to leave "The Blind Side" apprised of power dynamics and the prejudices that were the subtext of Michael Oher's trajectory, or incensed that this kind of racism and classism still persists. Nor are audiences able to get a picture of who Oher really is — a complex person, to be sure, who has experienced life on different sides of the privilege spectrum.
Instead, they get to walk away feeling moved by a paternalistic tale that acknowledges white, middle-class values as the only kind of values worth having, and celebrates the poor and the people of color who aspire to them.
Kristi Ceccarossi is from a working class, Italian-American family north of Boston. She works as the communications coordinator of the Program on Inequality and the Common Good at the Institute for Policy Studies.

subset: commoncrawl


text:
- Research article
- Open Access
- Published:
The dimerization interface in VraR is essential for induction of the cell wall stress response in Staphylococcus aureus: a potential druggable target
BMC Microbiology volume 19, Article number: 153 (2019)
2086 Accesses
10 Citations
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Staphylococcus aureus remains a medical challenge in the treatment of bacterial infections. It has acquired resistance to commonly used antibiotics, and to those considered to be the last weapons in treating staphylococcal infections, such as vancomycin. Studies have revealed that S. aureus is capable of mounting a rapid response to antibiotics that target cell wall peptidoglycan biosynthesis, such as β-lactams and vancomycin. The two-component system VraSR has been linked to the coordination of this response. VraS is a histidine kinase that undergoes autophosphorylation in the presence of signals elicited upon cell wall damage and it then transfers its phosphoryl group to VraR. VraR is a response regulator protein that functions as a transcription factor. Phosphorylation of VraR leads to its dimerization, which is required for optimum binding to its target promoters. Two-component systems have been targeted for the development of antibacterial agents. Deletion of the vraS or vraR gene has been shown to re-sensitize S. aureus to β-lactams and vancomycin.
In this study, we explored perturbation of the VraR phosphorylation-induced activation as a means to inhibit the VraSR-mediated signal transduction pathway. We show that dimerization of VraR is essential for the phosphorylation-induced activation of VraR. A single point mutation in the dimerization interface of VraR, in which Met13 was replaced by Ala, led to the inability of VraR to dimerize and to bind optimally to the target promoter. The consequences of these in vitro molecular deficiencies are equally dramatic in vivo. Complementation of a vraR deletion S. aureus strain with the vraRM13Ala mutant gene failed to induce the cell wall stress response.
This study highlights the potential of targeting the phosphorylation-induced dimerization of VraR to disrupt the S. aureus cell wall stress response and in turn to re-sensitize S. aureus to β-lactams and vancomycin.
Two-component signaling systems (TCSs) are prevalent in bacteria [1, 2]. They enable coupling of a diverse array of adaptive responses to environmental stimuli including antibiotic stress [1, 3]. Their absence in high eukaryotic systems makes them a prime target for development of novel antimicrobial agents [4]. Signaling via TCSs is based on a conserved phosphor transfer process between a histidine kinase (HK) and a response regulator protein (RR) (Fig. 1). The output response is controlled by the RR, which plays the role of a phosphorylation-activated switch [5]. About 60% of all identified RRs act as transcription factors [5].
The vancomycin-resistance-associated response regulator protein, VraR, and its cognate histidine kinase, VraS, constitute a two-component system in Staphylococcus aureus (Fig. 1a) that has been shown to mediate resistance to β-lactams and vancomycin [6]. VraSR has gained attention as a potential target in encountering the antibiotic resistance in multiple antibiotic resistant S. aureus strains such as MRSA. Deletions within the vraSR operon re-sensitize MRSA strains to β-lactams and vancomycin [7,8,9,10,11]. The VraSR-mediated signal transduction pathway is activated by the exposure of S. aureus to antibiotics and gene deletions that inhibit cell wall synthesis [6, 12, 13]. Cell wall damage in S. aureus leads to the VraR-dependent regulation of more than 40 genes [6]. However, the precise nature of the signal(s) that turns on the VraSR-mediated signaling pathway is not known. Both VraS and VraR are multidomain proteins (Fig. 1a). VraS consists of an N-terminal transmembrane domain, which, in a number of histidine kinases, is involved in sensing the extracellular stimuli, and a C-terminal core domain that harbors the dimerization interface, the conserved histidine residue, and the ATP-binding domain. VraR consist of a conserved N-terminal domain that harbors the phosphorylation site, referred to as the regulator domain (RD), and a variable C-terminal domain that interacts with DNA, referred to as the effector domain (ED) (Fig. 1a). Upon cellular stress, VraS is activated through an autophosphorylation event (t1/2 ~ 10 min) [12, 14]. Subsequently, the histidine kinase transfers the phosphoryl group, almost instantaneously (t1/2 ~ 10 s), onto VraR [14]. Upon its phosphorylation, VraR dimerizes at the N-terminal domain. Phosphorylation-induced dimerization is shown to expand and enhance the VraR binding to its own promoter (Fig. 1b) [14,15,16], and, consequently, to increase the expression of the vraSR operon [17]. Higher vraR expression leads to modulation of as many as 40 genes, which ultimately constitute the S. aureus response to cell wall damage. The VraSR-mediated signal transduction is reset in the absence of stress through the phosphatase activity of VraS toward VraR [14].
The events that lead to the phosphorylation-induced dimerization of VraR have been elucidated by hydrogen-deuterium mass spectrometry and X-ray crystallography [16, 18]. These studies showed occurrence of structural rearrangement in the VraR regulator domain in the phosphorylated dimeric species. In particular, the crystal structure showed that these structural rearrangements ultimately are associated with the formation of a hydrophobic pocket that wraps snugly the side chain of a methionine residue, Met13, which protrudes from an α-helix region (α1 helix) of the opposing protomer in the dimer complex (Fig. 2) [16]. This discovery provided a strategy for the targeting of two-component signal transduction pathways, that of inhibition of the phosphorylation-induced dimerization of RR.
VraR belongs to the NarL/FixJ family of RR, which use the helix-turn-helix motif to bind to DNA. The dimerization interface found in VraR is seen in a subset of this family of proteins, for example in Escherichia coli UhpA [19], a response regulator that regulates sugar uptake, Mycobacterium tuberculosis NarL [20], S. aureus LuxR (PDB ID 3B2N), a response regulator implicated in regulation of cell-density, Bacillus subtillis DesR [21], and Enterococcus faecium LiaR, a response regulator involved in daptomycin induced cell wall stress response [22]. The response regulator proteins that belong to the OmpR/PhoB family of proteins, which use a winged helix-turn-helix to target DNA, implicate the α4-β5-α5 interface for dimerization [23].
In this study, we investigated the role of Met13 in the phosphorylation-induced dimerization and binding of VraR to its own promoter. Our in vitro and in vivo studies revealed the profound effect that this single amino acid residue has on the cell-wall antibiotic stress response in S. aureus. In addition, they validated the significance of inhibition of the phosphorylation-induced activation of VraR in re-sensitizing S. aureus to β-lactams and vancomycin.
Characterization of the VraRM13A variant
The VraRM13A variant was purified to homogeneity as described before [14]. To assess the impact of substitution of Met13 by Ala on the overall secondary structures and the structural integrity of VraR, we recorded the CD spectrum of the VraRM13A variant and its thermal melting, respectively, and compared these data against the wild-type protein CD spectrum and the thermal melting. The CD spectra of VraR and VraRM13A were similar, indicating that the substitution of Met by Ala did not have a deleterious effect on the secondary structural features of the protein (see Additional file 1: Figure S1A). This outcome was to be expected as Met-13 residue sits on an α-helix (α1) and its substitute, alanine, is a strong α-helix former [24]. The thermal melting studies also showed that the stability of the variant protein was similar to that of the wild-type protein, inferring that the overall integrity of the protein was not affected (see Additional file 1: Figure S1B).
Phosphorylation of VraRM13A
Phosphorylation of VraR and VraRM13A by a small molecule donor such as acetyl phosphate was assessed by Phos-tag™ SDS-PAGE: the un-phosphorylated and phosphorylated protein species were separated by the SDS-PAGE as a result of the phosphorylated species mobility being slowed down by the Phostag reagent in the SDS-PAGE (Fig. 3). The protein bands, stained with coomassie blue, were quantified by ImageJ (NIH). These experiments showed that after 45 min of incubation the VraRM13A variant underwent about 10% phosphorylation compared to 73% of phosphorylation measured for VraR.
Oligomerization state of VraRM13A
Earlier studies on VraR have shown that phosphorylation drives the protein, otherwise a monomer, toward dimerization [14]. The Met-13 residue was identified as one of the residues that participated intimately in dimerization (Fig. 2) [16]. Hence, reduction of the methionine side chain to that of a methyl group should have an impact on the dimerization. The native-PAGE experiments were performed to investigate the protein dimerization ability after attempting the phosphorylation. The VraRM13A variant, subjected to the phosphorylation, did not form a dimer at 10 μM, 20 μM or 30 μM (or 40 μM, data not shown), unlike VraR (Fig. 4).
Analysis of the DNA-binding activity of VraR
To evaluate the DNA-binding activity of VraRM13A, we performed DNase I footprinting experiments. Previous studies showed that unphosphorylated VraR binds to the region − 50 to − 75 (namely R1) of the vrasR promoter (PVraSR) while phosphorylated VraR (VraR-P) binds to an additional site of the promoter, the region − 46 to − 26 (namely R2), (Fig. 1b) [15]. The DNase I footprinting experiments carried out with VraRM13A showed that unphosphorylated and acetyl phosphate-treated VraRM13A bound to PVraSR in the same way as unphosphorylated VraR (Fig. 5); only the R1 was occupied by these proteins, not the R2 site. These results are in agreement with the outcome of the phosphorylation and oligomerization experiments, which showed that VraRM13A subjected to phosphorylation with acetyl phosphate failed to undergo quantitative phosphorylation and any measurable dimerization. We analyzed the band intensities of three of the DNA bands observed in the VraR-protected region of PVraSR by ImageJ, and compared the DNA binding affinity of VraRM13A to that of VraR (see Additional file 2: Figure S2). The binding isotherms show that binding of VraRM13A to the R1 site of PVraSR was weaker than binding of VraR. We had shown earlier that the VraR DNA-binding sequence consists of two in-tandem sequences which in R1 are very close to each other, separated by one nucleotide, and as such they are likely to promote dimerization of VraR in this site [17]. The weaker DNA-binding observed for VraRM13A, in comparison to VraR, could be due to the inability of VraRM13A to engage into a dimer at the R1 site.
Activation of the vraR regulon in the presence of vraRM13A
To assess in vivo the effect of the Met13 to Ala substitution on the activity of VraR, we generated the PvraSR::vraRM13A construct and cloned it into the shuttle vector pMK4 [25]. The vector pMK4::PvraSRvraRM13A complemented the RN4220ΔvraR strain well as assessed by measuring the expression levels of vraR in the absence and presence of oxacillin using RT-qPCR (data not shown).
In the current study, we selected a few genes from the vraR regulon to assess the effect of the Met to Ala substitution into the transcriptional activity of VraR. These genes were fmtA, sgtB and pbpB [6]. Expressions of these genes in the absence of oxacillin were compared to their respective gene expressions upon exposure of RN4220ΔvraR(pMK4::PvraSRvraRM13A) to sub-inhibitory concentration of oxacillin, for 30 min (data not shown). In all the cases, the complementation of RN4220ΔvraR with vraRM13A variant failed to increase gene expression of vraR, fmtA, sgtB and pbpB in response to oxacillin. The lack of upregulation of the above genes in the presence of oxacillin is very likely due to the failure of vraRM13A to be induced by oxacillin in the RN4220ΔvraR(pMK4::PvraSRvraRM13A) strain.
Substitution of a single residue in VraR, Met13 to Ala, led to the inability of VraR to undergo quantitative phosphorylation by small molecule donors such as acetyl phosphate and to dimerize in vitro. In vivo, complementation of the RN4220ΔvraR strain with vraRM13A did not lead to a response to oxacillin-induced stress. We had earlier suggested that the dimerization domain in VraR is located at the regulatory domain [14]. The X-ray crystal structure of the VraR dimer showed that the dimerization interface in the active VraR species (BeF3−-VraR bound species) is located at the regulator domain of VraR [16]. Peculiarly, at the center of this dimerization interface was the side chain of Met13, sitting on the α1 helix and snugly fitted into a hydrophobic pocket formed on the opposing molecule of VraR in the dimer [16]. In this study, we show that substitution of Met13 with Ala does not have measurable effects on the overall VraR structural integrity as assessed by CD. Thus, any impact on the activity of VraMet13Ala was attributed to the lack of the methionine-13 side chain.
Autophosphorylation of VraRM13A by a small molecule phosphodonor such as acetyl phosphate was about 7-fold less compared to the wild-type protein. We attributed this outcome to the inability of VraRM13A to dimerize in the absence of the methionine sidechain. Dimerization is considered to be a process that drives forward phosphorylation of RR [26]. The aspartyl phosphate species carry a high negative free energy and dimerization can channel that energy in the form of conformational changes in the protein. These conformational changes, which mark activation of RR, facilitate binding of RR to itself (oligomerization) and/or the target DNA sequence or other proteins [26]. So if dimerization is impeded, the phosphorylation of RR will also be affected (Fig. 6). This explains the slower phosphorylation rate of VraM13A and highlights the significance of the Met-13 residue in dimerization of VraR.
Phosphorylation of VraR is required for upregulation of its own operon, the vraSR operon [15, 17]. We had shown earlier that phosphorylation-induced dimerization of VraR is required to bind to a lesser conserved DNA-binding site on the vraSR promoter, namely the R2 site (Fig. 1b); while a conserved DNA-binding site, namely the R1 site, in the same promoter, can recruit the unphosphorylated VraR protein (Fig. 1b). In the case of VraRM13A, the DNase I footprinting experiments showed that subjection of this variant protein to phosphorylation did not lead to binding to the R2 site even at protein concentrations as high as 50 μM. However, the conserved DNA-binding site, the R1 site, was occupied by VraRM13A. This result confirms our earlier work that dimerization of VraR is required for recognition of a DNA-binding site that is less conserved [17]. The significance of dimerization in gene activation by VraR was corroborated by the RT-qPCR experiments. Indeed, the wild-type vraR gene was able to complement the RN4220ΔvraR strain, and mediate the vraR regulon in the presence of oxacillin, as indicated by the increased in the expression levels of the fmtA, sgtB and pbpB genes. However, complementation of the RN4220ΔvraR strain with vraRM13A failed to mediate the vraR regulon in the presence of oxacillin, as indicated by the lack thereof of upregulation of the above genes. This observation is very likely due to the lack of upregulation of the vraSR operon in the presence of oxacillin. VraR is known to regulate its own operon, hence if VraRM13A is not able to bind to its own promoter optimally it will not lead to higher expression levels of vraR in the presence of stress, and subsequently VraR cannot regulate its target genes.
The impacts of substitution of Met13 with Ala on the phosphorylation rate, dimerization and gene activation by VraR demonstrates the significance of this residue in driving the phosphorylation-induced activation of VraR, and hence cell wall stress response in S. aureus. As such, targeting the dimerization interface in VraR, and possibly in other response regulators of the NarL/FixJ family, through small molecules is a strategy that can efficiently disrupt the signal transduction mechanism of bacterial response to their respective stimuli. In addition, due to their conservative nature, inhibition of RR offers a global strategy in targeting a wide range of pathogens. Furthermore, RRs offer a unique and a direct site of inhibition in TCSs; in the past, inhibition of HK did not lead to inactivation of RR due to the ability of RR to cross-talk with other phosphate-donors in the cell and by-pass the histidine kinase [4].
Chemicals, growth media, and antibiotics were purchased from Thermo-Fisher (Whitby, Canada) or Sigma (Oakville, Canada), unless otherwise stated. Escherichia coli strains, such as NovaBlue and BL21(DE3), and other expression plasmids were purchased from EMD4 Biosciences (New Jersey, USA). All the primers were purchased from Sigma (Oakville, Canada). Chromatography columns and media were bought from GE Healthcare (Mississauga, ON, CA). Radioactive [γ-32P]-ATP was purchased from Perkin Elmer (Life and Analytical Sciences, Waltham, MA, USA).
Generation of the vraRM13A variant, production and purification of the VraRM13A protein
The vraRM13A variant was generated through site directed mutagenesis using the QuikChange Site-Directed Mutagenesis kit (Agilent). The nucleic acid sequences of the mutagenic primers used in these experiments were as follows: Dir: 5′-GTGGATGATCATGAAGCGGTACGTAT AG, Rev.: 5′-CTATACGTACCGCTTCATGATCATCCAC. The pET26b::vraR vector [14] was amplified using the above mutagenic primer pair as suggested by the vendor. The resulting amplicon was subjected to digestion by DpnI and then used to transform E. coli NovaBlue. Mutation was confirmed by DNA sequencing at the Centre for Applied genomics (TCAG) facilities (Sick Kid's Hospital). The mutated pET26b::vraRM13A vector was introduced into the expression host E. coli BL21(DE3). The induction, isolation and purification of VraRM13A variant were carried out similarly to that of the wild-type protein, and as described by [14].
Circular dichroism (CD) spectroscopy and thermal melting of VraRM13A
The CD spectrum of the target protein was carried out by preparing a protein solution of 10 μM in 20 mM Tris, pH 7.0 buffer, and supplemented with 5 mM MgCl2. The CD spectrum was recorded from 200 to 260 nm in a Jasco J-810 instrument (0.1 cm path length cuvette). To investigate the effect of the mutation on the overall structure of the protein, the thermal melting of the protein was carried out by monitoring the CD signal at 222 nm from 20 °C to 100 °C in a Jasco J-810 instrument.
Phosphorylation of VraR by lithium potassium acetyl phosphate
Phosphorylation of VraR, or VraM13A variant (10 μM or 20 μM), was carried out with 50 mM acetyl phosphate in the phosphorylation buffer (1X PB: 50 mM Tris, pH 7.4 buffer, 50 mM KCl, 20 mM MgCl2). The reaction mixtures were incubated for up to 1 h at 37 °C. The extent of protein phosphorylation was evaluated by Phos-tag™ 12.5% SDS polyacrylamide gel electrophoresis (PAGE) [27].
Examination of the oligomerization state of VraR and VraM13A by native-PAGE
Dimerization of VraR and its variant upon phosphorylation was examined by native-PAGE. Briefly, the protein at concentrations 10 μM, 20 μM, and 30 μM was phosphorylated as described above and samples were resolved into a 10% native-PAGE at 4 °C.
DNase I footprinting assay
To perform this assay the region − 121 to + 26 of the vraSR promoter (PvraSR) was amplified by PCR using a 32P -5′-end direct primer Dir: 5′-ACGAAGCTTGGTCCGATTTTAACGAC AAAAATTG-3′ and a reverse primer Rev: 5′-TGAAATGACGCATTGATTGTGTTC-3′ [15]. The amplicon (2 ng) was incubated with VraR, phosphorylated VraR (VraR-P), VraRM13A and VraRM13A subjected to phosphorylation (VraRM13A/P) at different concentrations (0, 2, 5, 25, and 50 μM) in the binding buffer (10 mM Tris, pH 7.5 buffer, 50 mM KCL, 1 mM DTT) supplemented with 10 mM MgCl2, 50% glycerol and 20 ng/μl Herring sperm DNA. Each binding reaction was subjected to DNase I for 2 min to digest the DNA and then the DNase I stop solution (1% SDS, 0.2 M NaCl, 20 mM EDTA, pH 8.0) to stop the reaction. Samples were resolved by an 8% polyacrylamide gel containing urea, and the DNA bands were visualized using a Typhoon Trio+ Variable Mode Imager (GE HealthCare).
Construction of the complementing vector, pMK4::PvraSR vraR
In these experiments, the vraSR promoter region from − 121 to + 150 (PvraSR) was fused to the vraR gene (PvraSR::vraR) and the entire sequence was cloned into the pMK4 shuttle vector in order to complement the S. aureus ΔvraR strain and assess the effect of the M13A substitution into the VraR activity in vivo, by RT-qPCR. Briefly, PvraSR was amplified using the primer pair Dir: 5'AGGAATTCGGTCCGATTTTAACGACAAAAATTG and Rev.: 5'CGGGATCCAC GTTCAACATAGTTCATAAC (the underlined regions represent the sequences of the restriction enzyme sites, EcoRI and BamHI respectively) and the vraR gene was amplified using the primer pair Dir: 5'CGGGATCCATGACGATTAAAGTATTGTTTG and Rev.: 5'GCGTCGACCTAT TGAATTAAATTATGTTG (the underlined regions represent the sequences of the restriction enzyme sites, BamHI and SalI respectively). The PvraSR::vraR construct was ligated into the pMK4 vector at the EcoRI and SalI restriction sites. The sequence of the construct was confirmed by DNA sequencing. The pMK4::PvraSRvraR plasmid was introduced to S. aureus RN4220ΔvraR competent cells by electroporation (2 kV, 2.5 ms) using Micropulser (Bio-Rad) and were grown on TSB agar supplemented by 10 μg/ml chloramphenicol. Introduction of the mutation on the vraR gene at the pMK4::PvraSRvraR plasmid was carried out by site-directed mutagenesis as outlined above.
RT-qPCR to investigate expression of the vraR regulon genes in the RN4220ΔvraR strain complemented with vraRM13A
An overnight seed culture of one of the RN4220 strains was prepared in TSB media supplemented by 10 μg/ml chloramphenicol. An aliquot of 160 μl of overnight seed culture were used to inoculate 16 ml TSB medium supplemented with 10 μg/ml chloramphenicol. The subsequent culture was incubated at 37 °C (200 rpm) up to an optical density at 600 nm (OD600) of 0.4. The culture was split into two 5 ml aliquots and one of them was treated with 10 μg/ml oxacillin which was incubated along with the control sample, namely the non-oxacillin treated sample, at 37 °C for 30 min. An aliquot of 1 ml growth culture was used to isolate RNA using the RNeasy Mini kit (Qiagen) as per vendor's instructions.
High Capacity RNA-to-cDNA kit (Life Technologies) was used to synthesize cDNA from 500 ng of DNase I treated RNA. The 16 s RNA gene was used as an internal control using designed primers: Dir-5'GCTAAGTGTTAGGGGGTTTCC and Rev-5'TTCAACCTTGCGGTCGTACT. The 20 μl reaction mixtures consisted of 25 ng of cDNA, 0.25 μM of each primer (accordingly designed to target specific gene, Table 1), and 10 μl of SYBRE SELECT Master Mix (Life Technologies). The Rotor-gene Q qRT-PCR cycler (Qiagen) was used to amplify the cDNA. The PCR conditions were as follows: First hold: 2 min at 50 °C, second hold:10 min for 95 °C, 40 cycles of 95 °C for 15 s, 60 °C for 30 s, and 72 °C for 30 min; and 72 °C for 10 min as a final extension step. The sequences of the primers that were used to probe the expression of genes such as vraR, fmtA, sgtB and pbpB are provided in Table 1.
Availability of data and materials
The datasets supporting the conclusions of this article are included within the article and/or additional supporting files, with the exception of the RT-qPCR data. The RT-qPCR data are available from the corresponding author on reasonable request.
Adenosine triphosphate
Circular dichroism
Electromobility shift assay
Polyacrylamide gel electrophoresis
Polymerase chain reaction
Reverse transcription
Sodium dodecyl sulphate
- VraR:
Vancomycin-resistance-associated response regulator protein
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Iwata Y, Satou K, Tsuzuku H, Furuichi K, Senda Y, Sakai-Takemori Y, et al. Down-regulation of the two-component system and cell-wall biosynthesis-related genes was associated with the reversion to daptomycin susceptibility in daptomycin non-susceptible methicillin-resistant Staphylococcus aureus. Eur J Clin Microbiol Infect Dis. 2017;36(10):1839–45.
Dengler V, Meier PS, Heusser R, Berger-Bachi B, McCallum N. Induction kinetics of the Staphylococcus aureus cell wall stress stimulon in response to different cell wall active antibiotics. BMC Microbiol. 2011;11:16.
Gardete S, Ludovice AM, Sobral RG, Filipe SR, de Lencastre H, Tomasz A. Role of murE in the expression of beta-lactam antibiotic resistance in Staphylococcus aureus. J Bacteriol. 2004;186(6):1705–13.
Belcheva A, Golemi-Kotra D. A close-up view of the VraSR two-component system. A mediator of Staphylococcus aureus response to cell wall damage. J Biol Chem. 2008;283(18):12354–64.
Belcheva A, Verma V, Golemi-Kotra D. DNA-binding activity of the vancomycin resistance associated regulator protein VraR and the role of phosphorylation in transcriptional regulation of the vraSR operon. Biochemistry. 2009;48(24):5592–601.
Leonard PG, Golemi-Kotra D, Stock AM. Phosphorylation-dependent conformational changes and domain rearrangements in Staphylococcus aureus VraR activation. Proc Natl Acad Sci U S A. 2013;110(21):8525–30.
Belcheva A, Verma V, Korenevsky A, Fridman M, Kumar K, Golemi-Kotra D. Roles of DNA sequence and sigma a factor in transcription of the vraSR operon. J Bacteriol. 2012;194(1):61–71.
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Natural Sciences and Engineering Research Council of Canada is responsible for funding the project; Grant ID: RGPIN-05829-2015. This grant funded the design and execution of the project, the analysis and interpretation of the data, and the writing of the manuscript.
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Additional files
Additional file 1:
CD spectra. CD spectra of VraR and VraRM13A, and their respective thermal melting graphs. (PDF 57 kb)
Additional file 2:
DNase I footprinting. Binding isotherms extracted from the DNase I footprinting experiments. (PDF 45 kb)
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Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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Cite this article
Tajbakhsh, G., Golemi-Kotra, D. The dimerization interface in VraR is essential for induction of the cell wall stress response in Staphylococcus aureus: a potential druggable target. BMC Microbiol 19, 153 (2019). https://doi.org/10.1186/s12866-019-1529-0
DOI: https://doi.org/10.1186/s12866-019-1529-0
- Two-component systems
- VraSR
- Cell wall stress stimulon
- Staphylococcus aureus


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September 2016
A bit of book blurb..
Stay Dead is the heart stopping sixth book in Jessie Keane's bestselling Annie Carter series. Annie Carter finally believes that life is good. She and Max are back together and she has a new and uncomplicated life sunning herself in Barbados. It's what she's always dreamed of. Then she gets the news that her old friend Dolly Farrell is dead, and suddenly she finds herself back in London and hunting down a murderer with only one thing on her mind ...revenge. But the hunter can so quickly become the hunted, and Annie has been keeping too many secrets. She's crossed and bettered a lot of people over the years, but this time the enemy is a lot closer to home and she may just have met her match.
My thoughts about the book..
This is the sixth book in the Annie Carter series and I sure that there are legions of this author's work who devour her books as soon as they hit the bookshelf. As this was my first book by this author and therefore my first foray into the complicated world that Annie Carter inhabits, it must be said that I struggled with the story. Not because the writing lets it down, far from it, the plot and characterisation are very good, as the author clearly knows how to target her book towards her audience, but for me, it was more that without the benefit of knowing the series so far, I couldn't quite grasp the hidden nuances which make up so much of the story's overall effect.
There's no doubt that the author writes at a cracking good pace. The story moves along well and the hint of trouble and the constant threat of danger ensures that the story sits comfortably within the gangland crime/thriller genre. If you are already a fan then I am sure that you will love to see where Anna and Max's story moves along too, however, if like me you are new to the Anna Carter series then it's probably better that you start at the very beginning, it seems like a very good place to start to get to know the fierce and feisty, Anna Carter.
Best Read with... Large cups of coffee, heavy on the caffeine..
About the Author
Jessie Keane is a top twenty bestselling author. She's lived both ends of the spectrum, and her fascination with London's underworld led her to write Dirty Game, followed by best sellers Black Widow, Scarlet Women, Jail Bird,The Make, Playing Dead, Nameless, Ruthless (the fourth book to feature Annie Carter) and Lawless. Jessie's books have sold more than 750,000 copies, and she is a top 20 bestselling author. She now lives in Hampshire.
My thanks to Katie at Macmillan for my review copy of Stay Dead.


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Crane & General has been providing haulage services from our Cheshire depot for over 50 years.
With our fleet of vehicles and trailers we can transport goods all over the UK, including the transportation of abnormal loads.
Current industries served include:
If you have any further questions about our transport service, please do not hesitate to contact us.
Any Enquiries?
Call On : 01477 500334
All-Terrains up to 90 tonnes.
Truck Cranes.
HIAB Vehicles.
Office Hours
0800-1630 Mon - Thur
0800-1530 Friday
Calls diverted outside of these hours and bank holidays


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Our hotel was born from a dream.
Make a villa with our style for quality accommodation, as happened in the early twentieth century.
The choice of furniture in every corner of the hotel, music, events, breakfast are all designed in detail.
Each check-in for us has the same goal: to immerse yourself in a holiday to make it a dream for you too!
Our service offers many opportunities.
In addition to the rich breakfast until 11.30, we work with all types of restaurants and beach resorts as well as having many relaxing areas including a solarium and massage service at the hotel.
Tell us your needs, we will think of everything!
One of the privileges of our hotel is definitely the location.
In the center of Cesenatico with direct access to the boulevard Carducci, which in the summer evenings become pedestrian, but at the same time facing the sea. The entrance is marked by a large shady garden, where shelter from the hot summer days. The ideal for enjoying a beach holiday in a very relaxing and quality atmosphere!


text: The research involved a structured observation real-time use study of six New Zealand government chief executives, and examined Mintzberg's 1973 theory of managerial roles in practice. Mintzberg's theory of ten management roles was found to be relevant in 1997, but with the addition of the risk manager role, which is essential in a changing environment. Similar patterns of work to those described by Mintzberg were found; that is, chief executives jobs were "activity characterised by brevity, variety, and fragmentation." The NZ chief executives appeared to have a reactive orientation, being driven by external events and stakeholder and spent over 65% of their day in meetings. They never gave direct orders; and never finished a task that they started without interruption. They worked long hours, took short lunch breaks and spent some of their time fire-fighting with other agencies or the media. They also spent most of their time in the leadership, role exhorting staff to do more with fewer resources, for the good of society. This time use study gives a snapshot of the chief executives' role in the NZ public service in 1997. It has implications for the recruitment, retention and development of chief executives in the public sector and will be of interest to their employers, staff, information technology specialists, professional developers and researchers of management. It illustrates Mintzberg's managerial roles in practice.


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Now you can see a few examples of how jewellery designing is carried out in our workshop, and how we present our plans to our dear customers – we also show how the final products look like in relation to the plans themselves.
Affianced Jewel Design
Now you can see a few examples of how jewellery designing is carried out in our workshop, and how we present our plans to our dear customers – we also show how the final products look like in relation to the plans themselves.
Pieces of this collection were made with chiselled, magical gemstones, using white gold and three small brilliants.
The collection uses Japanese kanji as its main source of inspiration: their main idea is creating the perfect balance and ratio. The items do not symbolize the exact characters, but they do embody their characteristics and put their positive energies to good use.
Our guests can personally experience the joy of cooperation whilst designing an item. Constant interactions, consultations and if required tryouts make this process exciting.
As a first step, all the needs are measured through a simple conversation. To be more precise, I'd like to label it as collecting inspiration. Every piece of information can be important. For example, when making an item for a specific event and for a specific set of clothes, then the event itself can be a crucial inspiring element. A community occurrence or a ball have different requirements than a family get-together, or a wedding.
The décolletage, fabric, style, color of the dress, even the hairstyle, everything that changes outward appearance in the slightest can be vital. When talking about a female collection, the first item to be designed is always the necklace. We match the earrings' style to it, but, naturally, the hair can be a determining factor, too. A decorative collier doesn't need a larger sized earring. A showier one can match a delicate necklace perfectly. Our guests' figure and personality is required to be known as well. For a swan-necked bride with a dress that has a deep décolletage, we encourage making a longer earring for larger, flashier necklaces as well. On a more curvaceous woman, eye-catching and curiously designed items look the best. A thinner, delicate piece of jewelry can easily accentuate a plumper figure, which is not the goal. For further details, we call on the showcase models of our earlier designs for help, or their photographs. Familiarizing with these, we can determine what sense of taste we need to be targeting with our designs. Gathering technical information happens at this time as well: sampling sizes, determining lengths and widths, etc. After this comes the second step, meaning hand-drawn sketches with the earlier composed details in mind.
Worrying about the more detailed development is pointless yet. Even the structural solutions are just in our minds, roughly thought out. We design a maximum of 3 of the best ideas we come up with, to show our clients. This means well-detailed hand-drawn and computer designed sketches. By now, these sketches are supposed to show all the structural solutions, and where the materials are holding together precisely; after all, this will be the "map" of the designing work process.
After this comes arguably the most complicated task; putting together the designing schedule, appointing the tryouts so that everything can go smoothly until the given deadline is at hand. Evaluating the material needs and stating the price belong into this group of tasks as well. Naturally, we elaborate on this question as well during the first consultation, so that we can remain close to the imagined budget, but by our experience, the budget accepted later on is always larger. During the next consultation, we await our guests with the prepared plans, schedule and price calculation.
By this time we make experimental details and parts, also we put together a showcase of the desired materials, so that the final outlook of the item can be visualized even better. The first tryout usually takes place during this time, too, which means trying the paper cut-out of the flattened jewelry design of the item. In the case of a necklace, we point out the stronger curves of the collarbone and the neckline arcs of the shoulders on the paper mold. This will help making the structure-ready necklace's tilts to be precise.
On the next tryout, we present the piece of jewelry in this form. By this time, the production is in that state that we are required to anchor the arcs on the item. This way, the necklace will fit perfectly on the décolletage and every arc of the neck. Meanwhile we discuss the actual questions, opinions and notices, taking note of all of those that will determine further work.
After this is done, the next step is the detailed development of the item. If a small difference occurs during the process, an additional consultation may need to be held in order to acquire perfection. The final act of the process is handing the finished item over to its rightful owner.
The done product has to fit three criteria: function, development, design. Function means how comfortable it is to wear, and how well it fits the event it was meant for. Development is a professional question. Only development of perfect quality is acceptable. Design means the creative thoughts, the proportion sense and the aesthetic. If this trinity is achieved, then we can call it a job well done.


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We are traveling back from Napa today. Sad face.
In the meantime, here I am last week all bright-eyed and bushy tailed.
Luckily this was a quick trip so no jet-lag, please.
Chanel glasses | Gap sweater | LC Lauren Conrad jumper c/o Kohls | Target bag | AE ($8) sandals c/o thredUP!


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Ireland may be moving to infringe on more than just a child's right to life. Now, the country's leaders are expecting religious organizations to perform life-destroying procedures that contradict their values.
Simon Harris, the Minister for Health in Ireland, recently stated that all publicly-funded health service providers in Ireland must provide legal health services, which includes "women's health services" like abortion.
In a "statement of the obvious," Harris tweeted July 25 that "conscientious objections is for individuals, not institutions."
His tweet insinuated that no organization will be exempt from performing abortions, not even Catholic hospitals.
"All publicly funded health services providers in State will be expected to provide legal health services — including women's health services," Harris said. He went on to state that he has set up a group chaired by Catherine Day to "examine how voluntary providers interact with [Ireland's] health service overall."
All publicly funded health services providers in State will be expected to provide legal health services- incl. women's health services. This should be a statement of the obvious!Conscientious objection is for individuals, not institutions. Excellent article by @ellenmcoyne 1/2
— Simon Harris TD (@SimonHarrisTD) July 25, 2018
I have also set up a group chaired by Catherine Day to examine how voluntary providers interact with our health service overall. Very important work and group due to report to me shortly. 2/2
— Simon Harris TD (@SimonHarrisTD) July 25, 2018
Harris' Twitter posts come in light of outcry from Ireland's Catholic bishops, who have drafted a code of ethics for Catholic hospitals in the nation. According to The Times, guidelines for up to 20 main hospitals connected with religious orders include a ban on most assisted reproduction procedures. The document also lists some things that could be expected from a Catholic hospital, including bans on contraception.
Keep up with the latest pro-life news and information on Twitter. Follow @LifeNewsHQ
The Irish bishops are preparing to meet for the World Meeting of Families in late August, where they will address the need to defend life. This meeting, which Pope Francis is scheduled to attend, also will discuss plans to establish a "Council for Life," which will "advise and advocate for the Catholic Church in Ireland on a consistent ethic of life and care for those most at risk." They have yet to address Harris' recent declaration of forcing Catholic hospitals to perform abortions.
On May 26, citizens of Ireland voted to repeal their Eighth Amendment, which protected unborn children and women from abortion. This came as a great shock to pro-lifers across the globe, as Ireland had been one of the safest places in the world for pregnant women, with one of the world's lowest maternal mortality rates.
Now, the fear is Ireland's abortion law will mirror Britain's, where one in every five pregnancies ends in abortion each year. In Britain, abortion is permitted until 24 weeks of pregnancy.
Irish lawmakers are considering a proposal that would legalize abortion for any reason up to 12 weeks and up to six months for a wide variety of reasons.
With more access to abortion and organizations forced to perform them against their will, this safe haven for the vulnerable will not be so safe anymore.


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Hugh Jackman treated for cancer
After honing his skills facing down Magneto, Dracula and Javert, Hugh Jackman recently took on his most deadly foe of all – cancer.
The Jacked Man, 45, took to Instagram yesterday to reveal that he is undergoing treatment for Basal-cell carcinoma (as far as we can tell, that's a sort of little bastard cancer that generally turns up on your face), after heading to the doctor at his wife's suggestion. He's also rocking a mad cute little plaster on his nose, which virtually makes the whole thing worthwhile.
Deb said to get the mark on my nose checked. Boy, was she right! I had a basil cell carcinoma. Please… http://t.co/DESCvGR7GX
— Hugh Jackman (@RealHughJackman) November 21, 2013
(In case you were wondering, we're the ones that spelt his cancer right.)
We're horrified to hear that TJM was feeling under the weather, but if anyone can kick cancer's arse then it's him. Nothing says 'no need for chemo' like adamantium, eh?


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Hawaiian Homes Commission Chair Issues Statement on Seeking Act 14 Private CounselPosted on Sep 13, 2019 in Department of Hawaiian Home Lands, Media Releases
September 13, 2019
Hawaiian Homes Commission Chair Issues Statement on Seeking Act 14 Private Counsel
(KAPOLEI, HI) – Hawaiian Homes Commission Chair William J. Aila, Jr. issued the following statement on Friday, September 13, 2019, on the Department of Hawaiian Home Lands (DHHL) request to hire private counsel to advise the Commission on the valuation of compensation through funds and land owed to DHHL, pursuant to Act 14.
"I have submitted a letter to Hawaii State Attorney General Clare E. Connors requesting that DHHL hire private counsel to advise the Hawaiian Homes Commission (HHC) on the valuation of compensation through funds and land owed to DHHL, pursuant to Act 14. The private counsel will assist the HHC in carrying out the purpose of the Hawaiian Homes Commission Act solely in the interests of Hawaiian home lands trust beneficiaries.
"The HCC was briefed by DHHL staff in January 2019 on the background of Act 14 (1995) and the status of outstanding issues, which include the transfer of over 1,328 acres into DHHL's inventory. Resolving these issues is in the best interest of beneficiaries and the trust
"In furtherance of Governor David Ige's commitment to finalizing the resolution of this matter, DHHL, the Department of Land and Natural Resources, and the Attorney General continue to work towards this goal."
The Department of Hawaiian Home Lands carries out Prince Jonah Kūhiō Kalanianaʻole's vision of rehabilitating native Hawaiians by returning them to the land. Established by U.S. Congress in 1921, with the passage of the Hawaiian Homes Commission Act, the Hawaiian homesteading program run by DHHL includes management of over 200,000 acres of land statewide with the specific purpose of developing and delivering homesteading.
Cedric Duarte
Information and Community Relations Officer
Department of Hawaiian Home Lands
(808) 620-9501
[email protected]


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MUZIK will provide you an order confirmation and then a shipping tracking confirmation by your originally provided email.
Why can't I find my MUZIK order confirmation or shipping email?
- Please check your various emails in case you are not sure which one was used.
- Please also check your Spam folder as sometimes these emails initially are filtered there by your email service provider.
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As example, for Gmail:
Unmarking SPAM - Legitimate mail is marked as spam in Mail
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- Select the message,
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Vicky Howard
11 Items
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Colorful Images adds this beautiful artistry to it's collection.
Vicky Howard is an artist who has successfully created a brand identity for her illustrations and designs. From the beauty found in nature to the whimsy of a snowman, Vicky uses her brushes and paints to bring life to her favorite subjects. You'll also see her designs in file folders, note cubes, and more.


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Thursday, October 13, 2011
Trick or Treat
12 comments:
Cute, cute halloween card, Karen. Love the color graphic boxes, the twine and button. Super Cute!ReplyDelete
Cindy Porter
Really cute! Love the stamps and the twine! :)AmyReplyDelete
I love your card!!!! Very cute! I need stamps!!!! Sometimes the Cricut is not enough!ReplyDelete
Great card Karen.Love the twine.ReplyDelete
Have a great weekend.
Jayne x
really cute card my friend. Hope you are well, your card arrived today and really brightened my day...ReplyDelete
Hugs to you and Thank you.
Fun Halloween card!! Very cute:) TFSReplyDelete
So cute! Love the patterned papers you used. The cat and spider images are adorable.ReplyDelete
This card is fantastic! I love the big orange button! It adds the perfect touch! Thanks for entering our challenge at Scrapalicious USA! Best of luck!ReplyDelete
Love your Card Karen! I have a surprise for you on my blog.ReplyDelete
Hugs, Eva :0)
This is so awesome, Karen! I love the way you used the twine and those stamps are just adorable! You are so creative with designing cards - LOVE this!! :)ReplyDelete
ADORABLE project! I love it:) Thanks for playing with us at ScrapaliciousUSA.ReplyDelete
Hugs, Kassidy
This card is perfect! I love the placement of the button on your card and thank you for joining us at Scrapalicious USA Inc.ReplyDelete
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OS Battle: Windows 8 versus Mac OS X Lion
The Mac versus PC debate is about to heat up again. Both Microsoft and Apple have next-generation operating systems in the works, and while both companies will borrow heavily from their respective mobile platforms, it's Microsoft that's taking the bigger gamble. So how well does Windows 8 stack up with OS X Lion, and does Windows 8 have what it takes to win over new users (and keep current users from switching)? We compare a few key features to find out.
As far as extreme makeovers go, it's Windows 8 that will be turning the most heads at the next party. Gone: the Start button, taskbar and traditional Windows desktop. In its place is a mosaic-like wall made up of multi-colored widgets/tiles. Each widget will display live information on individual apps, giving the homescreen a grid-like appearance. (Alternatively, users will still be able to revert to the traditional Windows desktop if desired.) We're intrigued by the look of the new OS, but we can't help but wonder how users will respond to the new desktop, which at least in previews appears to be text-heavy.
Apple's home screen, on the other hand, won't undergo any jolting changes. Instead, a new Launchpad feature will swap your traditional home screen for a full-screen view of all of your apps (very similar to the home screen of an iPad or iPhone/iPod touch). Navigating multiple desktops will also be smoother thanks to the new Mission Control feature, which gives you a bird's eye view of every open app and window on your Mac.
Winner: Lion.
Microsoft's new UI will require that everyone reacquaint themselves with Windows. We're not sure a complete overhaul is the best approach for any OS.
Gesture support
Apple has supported multi-touch gestures on its laptops for quite some time, and Lion will expand on that support with new gestures it claims are more fluid and intuitive. These gestures will work on a Mac laptop, but desktops will require Apple's Magic Trackpad.
Microsoft is approaching gesture support differently. Since Windows 8 was designed with both desktops and tablets in mind, it'll feature native touchscreen support, so you'll be able to navigate Windows 8 on your desktop the same as you would on a tablet. The catch is, you'll need a touch-sensitive monitor to take advantage of these gestures. For those who don't own such a monitor, Microsoft assures us Windows 8 will work perfectly fine with a keyboard and mouse.
Winner: Lion
Microsoft is blurring the line between its desktop OS and tablet OS. However, we believe people use devices differently. We like our tablet OS to remain on tablets. In addition, not everyone owns (or can afford to upgrade to) a touchscreen monitor.
App Store
Despite Apple's best efforts to trademark the use of the term "App Store," Microsoft's next OS will have a built-in app store of some kind. However, Microsoft is being tight-lipped with details of its forthcoming storefront. In the meantime, Apple's App Store (which is already available on OS X Snow Leopard) will be more tightly integrated with Lion. In fact, Apple will only sell Lion as a download via the App Store.
Winner: Lion
Microsoft has been slow in integrating a digital storefront into its OS. Even with the announcement of Windows 8, Microsoft is still not sharing many details. This, in turn, has given Apple a solid head start in the App Store race.
Based on its previews, Windows 8 will be a radical departure from the Windows OS we've all grown used to. And while many features can change in the next few months, we can't help but feel that Windows 8 is being designed more for tablets than it is for desktops. Apple, on the other hand, is taking the safe route with Mac OS X Lion, bringing some of the best features of iOS 4 straight to the desktop — a move that many people already expected, and one that we think is closer to the future of desktop computing.


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By Luke Smith and Ruairi O'Connor
Changing your country and lifestyle is always a difficult upheaval, even if it is for a better life. However, how does it feel when, fifty years after coming over to the UK, you're told you're not really a citizen at all? Worse still, you never have been…
In the wake of the recent Windrush scandal, in which the government has threatened to deport hundreds of West Indians, we spoke to Herdle White, the longest serving Afro-Caribbean DJ in the BBC, about his experiences and reaction.
The Empire Windrush ship carried thousands of people from the Caribbean islands in 1948 to England, invited to settle here to help Britain tackle huge post-war labour shortages.
But it has now emerged documents such as landing cards proving their British citizenship were destroyed by the Government, making it harder for many to battle against moves being made for their deportation.
Herdle White has lived in Leicester since 1959 when he came over to the UK from Jamaica, working at the BBC for fifty years. He began work for BBC Radio Leicester doing a five minute Caribbean roundup, which has evolved into The Sound Of The Caribbean.
"I came over here in 1959 at a time when Britain was asking people from the Commonwealth, and especially the Caribbean islands, to come over. After World War Two there was a huge shortage of labour in this country, so people came over to plug the gaps.
"There were actually adverts in the Caribbean papers begging people to come over here to work, so lots of people came over, and lots of people came over during the World War, then went back to the Caribbean when it ended, and they ended up coming back over here too!
"In those days, everybody arrived with a British passport, you were a British citizen, and you were coming to the motherland to come and help the mother getting out of their difficulties, so you came with the British passport with no idea that in modern day you would be faced with deportation and all of the hassle that our people going through.
"When Jamaica became independent in 1962, I decided that I'd lived here long enough to take on British citizenship, which lots of people did do. It didn't enter anybody's mind that fifty years on you would be told that you're not British. It is very, very distressing for lots of people.
"Not in your wildest dream would you have thought you'd be in this situation and I cannot understand why people should be going through all of this, they've lived in the country for fifty or sixty years, what's the point in putting people through all of this now!
"People now can't get NHS treatment, can't get any benefits, you pay your tax and you can't collect your pension, all because of some missing paperwork, and I think this is absolutely outrageous and most of the Caribbean people I've spoken to are really angry.
"I spoke to some people on my radio show on Saturday night, and it was an Afro-Caribbean woman who used to live in Leicester who now lives in Jamaica, so I asked her what people were feeling over there and she said that people were mad as hell. It's a racist attitude and totally wrong.
"They're telling us that you've got to go back to a country where you now don't know anybody and you've got all your family over here, children, wives, grandchildren, great grandchildren even!
"I believe that the ten thousand or so missing bits of paperwork that have brought about this issue were destroyed deliberately. I also believe that because of the Eastern European migrants coming in, the government now realise that there are maybe too many people in the country and that they think the Caribbean people will be a soft touch so they thought "ah well we'll get rid of those 50,000 people that came over here" and then they can say that they've brought down immigration."
The integrity of the decisions made has also been questioned because many of the members of the Windrush generation are of retirement age and are collecting their pension, so it has been suggested it could be an attempt to bring in younger immigrants with an ability to work a trade and pay taxes, rather than collect pensions and retire.
"With all these things happening and still most of the MPs don't understand what is going on, and it's totally wrong," Herdle added.
"It's not like we came over here illegally, we were actually asked to come over and work, and now they want to send us back! We have a British passport to prove we're citizens and now we're being told that we're not and it's wrong. For me as a black person, I automatically think that the reason they're doing this is because I'm black, and that's the way people think."
Whilst Herdle is part of the Windrush generation, and whilst his generation of people are currently having their citizenship questioned, he insisted that Britain is still his country and will always feel like home.
"Although people are obviously very angry about this situation, we still feel like this is our country. They wouldn't want to go back to the Caribbean. You live sixty years of your life here and it's wrong to send people back.
"The Prime Minister has stated that all of the people affected by Windrush will be compensated, so it is now the responsibility of the Caribbean governments to make sure that the message that is given is passed on and implemented."


text: "An endeavor to create a product that encompasses the individuality and uniqueness that can only be found in natural surroundings. A desire to experiment with design, materials, and aesthetics in the search for something that works. Free from passing trends, the brand's focus remains that of its roots, innovation and creation."


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Late Monday, it was revealed acting Attorney General Sally Yates, an Obama appointee, had refused to enforce President Trump's new immigration policies and she had instructed Department of Justice lawyers to refrain from following through with President Trump's order.
Following this refusal, the Trump administration dismissed Yates, and replaced her with Dana Boente, US attorney for the Eastern District of Virginia.
Along with Yates' dismissal was a statement from the White House saying that Yates "betrayed the Department of Justice by refusing to enforce a legal order designed to protect the citizens of the United States," and that she "is an Obama appointee who is weak on borders and very weak on illegal immigration."
Boente will serve as acting attorney general until Senator Jeff Sessions, Trump's nominee, is confirmed by the Senate, The Blaze reports.


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45" x 30" (114.3 x 76 cm) Digital C print, unframed
"Paradise series is a convention. It is something unreachable, like a parallel utopia where perfection and beauty are prioritized. I want to explore the idea of the subjectivity of a paradise and how this "ideal place" can be filled with our visions projected upon it. They are different for each of us. Paradise can be related to memory, childhood, and dreams. As children, we imagined it to be a fantastic place, full of illusions. When we grow up and become less innocent and more skeptic, this convention survives, although much of that place has been abandoned and replaced with new ideals, like professional achievements, endless love, success, happiness, beauty, youth, etc. " - Lucia
Artist is influenced by Henry Matisse, Frida Kahlo, Irving Penn, Richard Avedon, Gustav Klimt
Born in Buenos Aires, Argentina in 1986. Artist has studied Theatre Costume Design at the Teatro Colon in Buenos Aires. Fainzilber studied Art Direction in Filmmaking at The University of Cine in San Telmo, Buenos Aires. After graduating in 2008, she has worked as a colorist in post-production and color-correcting movies in a Visual Mastering Studio before moving to New York to study at the International Center of Photography. After finishing The One-Year Certificate Program at ICP, she assisted to various fashion photographers in NYC as well as pursuing her own editorial work for international magazines like Dossier and campaigns for international and local brands. In November 2017 Lucia will exhibit at the Paris Photo in France. She is awarded the First Prize from the International Fine Art Photography Awards.
01/ 2011 "Moment of Recognition", curated by Amy Arbus. Rita.K. Hillman Education Gallery at the International Center of Photography, New York.
06/ 2012 "My Truth, Your Truth", curated by Alison Morley and Marina Berio. Rita.K. Hillman Education Gallery at the International Center of Photography, New York.
06/2014 Solo Show "And Spring Again", curated by Karen Grimson, Argentine Consulate in New York.
10/2014 Solo Show "Somewear", at Praxis Art Gallery in New York City. Participating in Art Miami 2014 and LA Art Fair 2015.
04/2016 Solo Show "Wild Flowers" at Praxis Art Gallery in New York City.
05/2016 Solo Show "And Spring Again" at The Argentine Embassy in Washington DC.
10/2016 Group Show "A Women's Thing" at FXFOWLE Studio in New York.
01/2017 Group Show "Angry Women" at Untitled Space in New York.
06/2017 Solo Exhibition "The Dress Code" at The Argentine Consulate in New York.
11/2017 Group show at Paris Photo in Paris, France (Winner of the International Fine Art Photography Award).


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Your hub for baritone news, tips, tricks, rare gear, and more.
Got a baritone you want to sell? Looking for a great deal on a used baritone or a hard to find extended-scale jem? Look no further. Private sellers are welcome to post links to a listing for their used baritone. You can also post a full description/add in the comments section if you like.
1/24/2016 11:24:23 am
Hi! I have a VERY RARE Larry Pogreba Baritone for sale at a very reasonable price. If you have any questions, please email me. FYI, I already have an offer so respond in a timely manner if interested. Thanks!
Joshua Jones
2/27/2016 02:03:41 pm
I have a PRS SE baritone I'm looking to sell, $500
Victor Tomkinson
3/10/2018 02:49:51 am
Hi, I have a 2007 Danelectro Longhorn Baritone in Bullion Gold, a little gig-worn but well looked after and in very good playing condition, looking for aound £450. Lovely guitar, I'm selling because it has no whammy and no way of installing one without serious modification. (I have two other baritones, so need to sell this one really)
Leave a Reply.


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Lush Green Services has been providing commercial and residential clients with various landscaping and maintenance services since 2003. As a family owned and operated business we focus on providing the very best landscape maintenance services in the area.
Lush Green Services offers the highest quality landscape maintenance service available in the area all backed by a 100% satisfaction guarantee. Call (970) 201-1967 to get our expert landscape maintenance services scheduled today.
Our landscape maintenance services offer customers pleasing results completed on time and on budget. We use the high quality products for lasting results and of course all of are equipment and personnel are insured, so you can rest assured that we have you covered. Our office staff is available to answer questions or schedule services for your at (970) 201-1967.
Many properties in the area have beautifully landscaped spaces which include lawns, trees, and shrubs as well as flower beds. Keeping all these spaces looking good and in good health year is a huge chore. We at Lush Green Services have been providing commercial and residential property owners high quality, customized landscape maintenance solutions since 2003. With a wealth of experience in the field we have the expertise, skill and resources to provide excellent shrub and flower bed maintenance services based on your specific needs.
Call or text our office staff during our office hours and we will be ready and happy to help.
Office Hours:
Monday 8:00am – 5:00pm
Tuesday 8:00am – 5:00pm
Wednesday 8:00am – 5:00pm
Thursday 8:00am – 5:00pm
Friday 8:00am – 5:00pm
Saturday & Sunday Closed
For a fast and free quote, fill out our form


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Sapphire Endure
by ROCOL
Original price $245.00 - Original price $26,800.00
Original price
$245.00 - $26,800.00
Current price $245.00
Premium Extreme Temperature Resistant Grease
- Extreme chemical and solvent resistance
- Resists caustic wash down
- Greatly extends lubrication intervals
- Excellent load carrying and anti-wear properties
- Approved by Rolls Royce for use on Harrier Pegasus Thrust Nozzle Drive Mechanism. Also approved for Submarine Clearance
- Extremely stable at high temperatures – temperature range of -25°C to +280°C
- NSF H1 Registered
- ISO 21469 Certified
- DETEX technology – x-ray and metal detectable plastic aerosol caps and actuators reduces risk of plastic contamination
- HALAL Certified
Extreme performance, perfluorinated polyether, extreme pressure grease fortified with PTFE. Designed to operate where conventional greases fail due to temperature or chemical attack.


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OPACJOB - Bib Usage Log recording "G" client type
- Article Type: General
- Product: Voyager
- Product Version: 7.0.4
Bug Report for Issue 16384-10321
Module(s): Opacjob
Server platform(s) affected: Tested on Solaris
PC OS (if applicable): N/A
Browser & version (if applicable): N/A
Release(s) replicated in: 7.2.1, 7.0.4, 7.0.1
Last version without bug (if applicable): 6.5.4
Expected results:
The client type used in the bib usage log should match one of the values in table 15-2 of the Technical Manual.
Actual results:
The client type used in the bib usage log for the OPAC is being recorded as "G" instead of "W"
Workflow implications: Customers may not be able to identify when a bib is being accessed by WebVoy?ge.
Replication steps:
1. Load /m1/voyager/xxxdb/ini/voyager.ini and set LOGBIBUSAGE=Y
2. Load the Cataloging client, pull up any record, and save it to the database
3. Load WebVoy?ge, and browse to the same record
4. Run Popacjob –j3 on the server, and use "today" as the parameter
5. Check the contents of bibuselgexport.dat. It should show an entry for Cataloging, and an entry for WebVoy?ge. The WebVoy?ge entry will show "G" for the client type
- Article last edited: 3/16/2015


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Premier Cyber Security Event Draws 1,300 Cyber Security Professionals to Capital Region to Discuss Latest Industry Trends
Governor Andrew M. Cuomo today announced that the 22nd Annual Cyber Security Conference, New York's premier cyber security event, begins today in Albany, bringing over 1,300 cyber security professionals from state and local government, academia, and the private sector to discuss emerging developments in the industry and best practices designed to improve the security of New York State. The conference, held at the Empire State Plaza through tomorrow, June 5, aligns with the Governor's longstanding commitment to protect New Yorkers and businesses from cyber threats and improve the safety and security of the State.
"The Annual Cyber Security Conference brings industry professionals from across the state together in one place, working toward the common goal to increase New York's cyber security infrastructure," Governor Cuomo said. "As we face increased cyber threats across the globe, this longstanding conference is more critical than ever to ensure we can continue to lead the nation to protect New Yorkers from devastating cybercrimes."
"We understand the importance of cyber security and continuing to work to combat cyber threats in New York," said Lieutenant Governor Kathy Hochul. "The annual Cyber Security Conference brings thousands of experts and professionals in the field to the Albany area to discuss best practices to improve security across the state. By bringing together the best and the brightest, we are fostering innovative solutions to ensure all New Yorkers' personal information is protected and kept safe."
The annual Conference is hosted by the New York State Office of Information Technology Services (ITS), in partnership with the University at Albany School of Business and the New York State Forum, Inc.
Senator Diane Savino said, "We've seen how cyber attacks can affect all levels of government and have a devastating impact on both small and large businesses. Cyber security has never been more important and this event is an important way to help share new ideas across the industry so that people are better protected. I thank Governor Cuomo for continuing to organize this event and look forward to working with those attending to ensure New Yorkers are protected from cyber attacks."
Assembly Member Clyde Vanel said, "We've made great strides by studying and adopting new tech under the guidance of New York State's forward thinking executive, Governor Andrew M. Cuomo. The 22nd Annual Cyber Security Conference is an opportunity to continue the debate and conversation, and to bring together industry leaders, cyber security experts, government and academia. We must assess and improve cyber security awareness in New York State and we must always continue to educate."
"In this digital age, cyber security is of paramount importance," said New York State Chief Information Officer Robert H. Samson. "Thanks to the leadership of Governor Cuomo, New York is well-prepared to address cyber threats. Industry partners at the New York State Cyber Security Conference help us keep pace with the trends in cyber security. We must all work together to keep New York safe from cyber threats."
New York State Chief Information Security Officer Deborah Snyder said, "Our Annual Cyber Security Conference helps New York State raise awareness about the importance of cyber security and educate our many partners on the most effective cyber security strategies to safeguard their organizations' information. We all have a role to play when it comes to cyber security. In New York, we are doing our part to strengthen the cyber workforce and protect New York's information, systems and critical infrastructure from cyber criminals."
Mario Musolino, Executive Director of The NYS Forum, Inc., said, "Every day there are countless cyber attacks aimed at government, industry and individuals. It is important that we understand what is happening and make sure that we are protecting our valuable assets. This conference is a unique opportunity for all of us to learn from each other regardless of our level of expertise."
"The UAlbany School of Business has a long history of supporting the NYS Cyber Security Conference and organizing the Annual Symposium on Information Assurance. We value this continued collaboration with the New York State Office of Information Technology Services and the resulting interaction of academics and professionals at the event," said Dr. Nilanjan Sen, Dean of the School of Business at the University at Albany."This conference is instrumental in improving awareness of challenges and new developments in the field to a broad audience. The school's strong focus on digital transformation requires that we not only dig deeper in the field but also share research, trends and challenges. We are thrilled to co-sponsor the premier cyber security event in the Northeast."
"We believe education and awareness are critical to address today's ever-increasing cyber threats, and for that reason it is a focus of our cybersecurity outreach to local governments," said Division of Homeland Security and Emergency Services Acting Commissioner Patrick Murphy. "The 22nd Annual Conference will address these efforts and other important initiatives. This annual event underscores the importance that stakeholders from across government, the private sector and academia must continue to work together to combat the increasing amount of cyber threats facing the public, business and government."
"With emerging technologies, law enforcement is constantly challenged by criminals online," said New York State Police Acting Superintendent Keith M. Corlett said. "Through this conference, we can boost cyber security awareness and collaborate with public and private partners, sharing information and strategies to help protect New Yorkers and meet new threats head-on."
"Education and information sharing by all levels of government and academia has benefitted New York State's cyber readiness," said New York State Board of Elections Co-Executive Director Robert A. Brehm. "This conference can increase cybersecurity awareness by updating us on emerging technology and trends."
The 2019 conference features more than 50 sessions, many of them including interactive discussions led by subject matter experts in government and the private sector. Google Cybersecurity Executive Michael Aiello delivered the opening keynote address entitled "Cyber Fragility in 2019." His address encouraged audience members to consider the approaches modern organizations are taking to reduce risk, build trust in technology, and analyze how emerging attacks may impact themselves and their organizations.
For more information on the 22nd Annual New York State Cyber Security Conference, visit the conference website at https://its.ny.gov/2019-


text: How Barb Carey Set out to Fill a Void for Women in the Sport of Fishing - 004 Posted by Angie Scott · February 13, 2018 In episode 4 of the It Pays to Fish podcast, you'll meet Barb Carey, founder of Wisconsin Women Fish and the Women Ice Angler Project. Barb grew up in the Upper Peninsula of Michigan and developed a love for the outdoors and fishing at an early age. When she went off to college, she drifted away from it, but eventually rediscovered her passion for fishing again. As she set out to learn the techniques to catch more fish, she noticed a huge void for female anglers in her area. Thus in 2003 Wisconsin Women Fish was born. With just a handful of ladies who shared the same passion for fishing, through word of mouth the group started to grow, and now 12 years later is 300 members strong, including members from all over the midwest and other parts of the country. Aside from social outings, the group also hosts educational clinics. From WI Women Fish, came the Women Ice Angler Project. Barb, once again, saw a need. There was a huge lack of female anglers being featured in product catalogs, promotional materials and other publications. WIAP is an annual media event to generate and showcase photos of real women ice fishing. They make a huge splash on social media using the hashtag #womenonice. Only in existence for four short years, WIAP has already made a huge impact in the ice fishing world and results are being realized. In this episode you'll learn: How Barb Carey set out to fix a gap she noticed in the fishing industry and turned it into a full-time job. How she uses writing and other communication skills to her advantage in multiple areas, including seeking out sponsorships and pro staff positions. About Wisconsin Women Fish and the Women Ice Angler Project, and how they've brought female anglers together from all over the midwest and other parts of the country. Additional Resources: WI Women Fish WI Women Fish on Facebook WI Women Fish on Instagram The Woman Ice Angler Project The Woman Ice Angler Project on Facebook The Woman Ice Angler Project on Instagram The Great Wild Radio Show This episode is brought to you by Mariner's Learning System. Go to www.marinerslearningsystem.com and enter coupon code: itpaystofish upon checkout to save 10% on any of their United States Coast Guard approved Captain's License programs! Right-click here to download this episode to your computer.


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Earn your Associate degree in business from a top college below. Even strong endeavors fail from lack of firm leadership. Just ask the ancient Athenians around 400 B.C. who, despite having overwhelming numbers, technology, and advantage, failed to conquer the island of Sicily due to hesitant and underprepared leaders. As a result, Athens epically lost their military and economic domination of the region. Or, more recently, just as the top executives at Enron, formerly recognized as one of the world's most innovative companies who illegally manipulated accounting statements and failed to ethically administer their business.
A business is only as strong as its leadership; and if that leadership has effective administration skills that can make decisions, coordinate employees, meet goals, and produce profit, then it is destined for enduring success. By getting your Associate Degree in Business Administration, you can begin to join this elite club of leadership.
Every business needs administration to ultimately find success and the better the business administrator, the more likely success will occur. One of the distinguishing elements of a business administrator is that they understanding and can make accurate decisions about a wide range of business aspects. Normally, individual employees develop a specialized expertise; however, a business administrator comprehends how to use these employees and their individual skills to make the overall business operation a fluid machine.
Business administration programs often focus on developing this wide skillset, including areas of training like accounting, marketing, leadership, information systems, economics, business law, and business strategy. When a company has an experienced and educated business administrator who can competently manage all of these diverse aspects of business, then that company will be a powerfully united whole that will easily distinguish itself in the marketplace. See the business administration degree overviews below from each school to get started.
In addition to administrating a business at the upper levels where you'll incorporate all of the above listed areas into your everyday decision making, many business administrators are also hired to lead one particular section of an organization. For example, larger organizations might require a talented business administrator to lead their accounting department, or their Human Resources department. In this scenario, you might find that your broad knowledge of a business's functions will serve you well when leading your particular area of a company toward a unified vision. Many business administrators also take on a career as an independent consultant, contracted by a particular company for a time to improve their efficiency, market presence, or other specified aspect.
Businesses are the commonly expected career area for a business administrator. After all, it's in the title of the degree. But business is not the only place where talented administrative leadership is needed. Education is also a vast area requiring leadership from superintendents to principals, from school board members to department chairs, education requires individuals with business administration backgrounds to successfully manage the vast array of operations a school performs. Plus, you can take your talents directly to the classroom yourself and become an instructor who teaches others what you know about business administration. Another non-business area that relies on strong leaders to manage resources is in all level of government. Every city has a mayor that staffs the city with managers, chiefs, officers, and all kinds of other posts all posts that, just like business, require individuals who can make informed decisions based on their broad education to ensure success towards a unified vision. Find top online associate of business administration degrees above to get started.
The Athenians had a poor general who misunderstood their position and led them to their defeat. But that's ancient history. Today, you have the advantage of obtaining an Associate's Degree in business administration, which will teach you the essential skills and strategies of business, and give you the perspective you need to be able to lead any business, any institution, any organization to their ultimate triumphant success.
When attending any of the business schools listed you'll cover a wide variety of business courses. These courses are designed to give you an overview of a real-world business environment. Some of the courses you'll cover include:
- Accounting (both business accounting and financial accounting
- Finance
- Human Resources
- Marketing/Advertising
- Management
- Small Business Management
- Corporate Supply Chains
- International Business
- Statistics
Business Management – As a business manager you'll work directly in operations for all sizes of businesses. You'll supervise workers and be in-charge of daily activities. As a manager you'll need to have a business focused skill set which includes the ability to work with people and solve decisions quickly.
Account Management – Account managers hand specific accounts or parts of a business. They can be responsible for business accounts working with clients or even oversee budgets. It is your responsibility in this role to grow the account either financially for your business or the client you work for.
Customer Service Managers – Most business administrators when they first graduate college will work in a customer service role. These calls centers can be for large corporations where you'll handle daily calls from customers.
Marketing Consultants – As a business major your Associate's degree in business administration can open up other avenues you might not have thought about including marketing. Most admin degrees will also train you in marketing. As a marketing consultant you can specialize in a particular side of business or marketing in order to help your clients. This could be areas in HR, advertising, hiring, auditing and more.
Auditor – As a business auditor you'll deal directly with corporate fraud or even waste. Companies are always looking for ways in how to reduce both and often this can be done on a consulting basis.
Sales Management – Learn to work with teams during your college training and work for a company.
Tuition – The top thing to research at any school is their tuition costs for the program. Your Associate's Degree in Business Administration should run as low as $150 per credit hour to $300. We have seen program in excess of $30,000 but on average they are $15,000. Your degree will be broken down into classes that are usually 3 credit hours long and you'll need 60 credit hours to graduate.
Online Vs. Campus – Depending on where you live you might not be able to find a school in your city that offers this degree. The great thing is we list plenty that are available 100% online. By taking your AA degree online you can actually save time and even complete it quicker than a traditional college. You won't have to drive to class and most books these days are fully online. Also by not attending a campus you can choose from any college in the county.
Feedback – We always recommend looking online and doing a quick search for the school to see if they have too much negative feedback. No school will be without some form of negative feedback but we want you to feel comfortable with who you are enrolling with. Try to avoid schools where when you search the entire first page is nothing but student complaints. If you would like some assistance feel free to send us an email.
AACSB- is the association to advanced collegiate schools of business. It is a national accreditation. Only 25% of all business schools are this accredited. - Updated January 2, 2019
ACBSP– The accreditation counsel for collegiate business schools and programs. Visited January 20, 2019
Chea– The council for higher education accreditation who accredites over 3000 colleges natwionwide. Added November 3, 2018


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This is simply put, the most adorable picture I have seen in years!
Date: 4/12/2018 Views: 10478 Favorites: 185 Comments: 10
A commissioned headshot of a cute girl turning into a cuter bunny girl.
Lookit those big floofy ears!
To add a comment, please sign in or create an account.
This is simply put, the most adorable picture I have seen in years!
Aww, thank you! I am immensely flattered.
gotta say, that's some wonderful coloring you've got going on
Thanks! It takes forever, but it looks great.
She is a very beautiful girl for a dinner.
I'm sure she would eat more than a salad.
Absolutely love this, 97/100
Very good! Would love to see more.
Thanks! I do have a whole page with much more over on FA.