converse uk Regulations for the reading ro

Regulations for the reading room for archives and rare books

All visits must be made by prior written appointment with the search room staff. On their first visit and in each subsequent calendar year, all readers are required to complete a registration form and produce converse uk proof of identity including their permanent address. each day.

All readers are required to complete a formal declaration that they will abide by these regulations and all other regulations for the use of and reproduction of mate converse uk converse uk g>rials held within the Special Collections. In addition, all readers are required to sign the visitors’ register each time they enter and each time they leave converse uk the reading room. Signing implies an undertaking to abide by these regulations and all others for the use of and reproduction of materials held within the Special Collections. Infringement will render a reader liable to exclusion and could result in contractual liabilities.

converse uk Regulation and a Touch of Corp

Regulation and a Touch of Corporate Responsibility Put More Green into IT Knowledge

The legions of fans who thronged to drop $200 on an iPhone 3G in July probably did not think much about the gadget fate when obsolescence eventually renders it scrap. But Apple definitely has been. Under pressure from tightening global anti pollution standards, threat of environmental lawsuits and more awareness of corporate responsibility, Apple and other technology firms are racing to place a stamp of environmental approval on their operations and products.

It easy to understand why, when one considers the electricity used to run data centers (and keep them air conditioned), the vast quantities of water and toxic chemicals used to make components and the mountain of electronics put out to pasture each year by consumers and businesses.

According to the International Association of Electronics Recyclers, about 400 million units of electronic junk, or is generated annually. Most of it is thought to reside in consumers closets and basements as consumers become more reticent to send their gadgets to landfills, but have no clue how to responsibly dispose of them.

hear people say, this green thing a fad? notes Christopher J. Lynch, director of the Wharton based Environmental Management Assistance Program. He says the last time environmental enthusiasm ran this high was in the early 1990s, when high oil prices from the first Gulf War roughly coincided with the 20 year anniversary of Earth Day. think this time around, it different, says Lynch, who also serves with Wharton and the University of Pennsylvania Initiative on Global and Environmental Leadership.

With even many former skeptics acknowledging that humans do increase greenhouse gas emissions, there seems to be a longer term focus on green initiatives, converse uk he suggests. As an adviser to private companies through Pennsylvania Small Business Development Centers, Lynch says more companies than ever are asking now about what they can do to green up their operations. Plus, he adds, dealing with higher energy costs.

Research firm Gartner estimates that at a minimum, 2% of global atmospheric carbon emissions can be traced to the information technology industry, because of the electricity consumed by PCs, servers, cooling systems, telecommunications and printers.

Changing the Corporate Culture

Such statistics aren lost on Bangalore based IT services firm Wipro Technologies. In June, the 96,000 employee firm launched a companywide initiative called Eco Eye. It calls upon every employee to consider how his or her everyday actions impact the environment. firmly believe business cannot be built at the cost of ecology, Wipro chairman Azim Premji said when announcing the plan in June. is not sustainable. Ecological sustainability will increasingly be the defining force for society and business globally. Wipro believes ecological sustainability is the right thing to do in fact, it is the only way forward.

Among other things, Wipro conducts screenings of Al Gore global warming documentary, An Inconvenient Truth, for employees; female employees are given burlap sacks to discourage the use of plastic bags and to have them serve as Ambassadors in their circles of social influence outside of the workplace.

objective is to create and spread awareness among employees and influence a gradual conversion to eco friendly behavior, says Swapnil Bhatnagar, a Wipro spokesman.

Using its products to reflect the corporate philosophy, the firm says it has or will launch green data centers; introduce a product called Greenware PCs, which it says is India first eco friendly line of personal computers; conduct Earth friendly product engineering; and start an e waste disposal service for electronics that have reached the end of their useful service lives.

Why do it? Lynch says companies seem to be acting for many reasons, but a large one is simply increased awareness. There a greater willingness of managers now to say, got to get on this green bandwagon. Until late last year, small businesses almost never sought out his office on their own, looking for help with efficiency. the past, compliance was a driver, but now we got more interest in green.

A Push from Europe

Helping to push this wave of eco enthusiasm along is a standard that most Americans have probably never heard of, but is as familiar as the Clean Air Act among people in technology circles and in Europe. It the European Union Restriction of Hazardous Substances directive, which applies to electrical and electronic products for sale in EU member countries. Known as RoHS, and pronounced or alternatively, converse uk the decree took effect in 2006 and forbids the use of lead, mercury, cadmium, hexavalent chromium and polybrominated flame retardants in electronic products such as computers.

Europe, there more of a willingness to move forward on environmental protection, says Matthew White, a business and public policy professor at Wharton. has been a whole lot more willing to pay the short term price. That price, he added, can include higher upfront costs of enacting more aggressive regulation and subjective costs such as aesthetic concerns about the placement of wind turbines.

The effects of the European rules have been far reaching.

The Telecommunications Industry of America, for instance, offers its members a service called EIA TRACK a global bill tracking service from the Electronics Industries Association that provides members a heads up on proposed environmental standards. Depending on a company revenue, annual subscriptions run from $3,000 to $17,500, for which the organization subcontracts an army of attorneys to monitor developing legislation. couldn do it on their own for anywhere near that, says Mike Snyder, a TIA spokesman. seeing that the industry is concerned with environmental matters, says Ellen Farmer, also of TIA. are heading in a more corporate responsible direction. When her book debuted in 2006, it exposed to the wider public some of the IT industry dirty laundry, such as e waste dumping, toxic contamination of water sources during manufacturing and other dangerous practices. actually been quite a bit of movement on the issues, Grossman says. people first started talking about this, none of the manufacturers thought producer responsibility would work. responsibility is the buzz term for electronics makers reclaiming their products once they are no longer wanted, instead of placing the burden on the consumer to find a recycling program. In Europe, it a requirement of doing business.

That important, Grossman says, because of the haphazard manner in which electronics have been disposed of in the past. computer will accept junked electronics from consumers, businesses and government agencies; this then gets sold to a wholesale operation, which ships the junk to China, India or another low cost labor country. At its destination, unprotected workers in squalid conditions break apart the computers, TVs, printers and other e waste and recover their valuable components. What can be salvaged such as arsenic and lead filled monitors is piled up and allowed to leach into the soil and the local water supply. The shock value of films such as the Basel Action Network Exporting Harm, which depicted such scenes, placed a spotlight on e waste.

Grossman says that when the film debuted in 2002 at a conference for electronics recyclers, gasps of surprise came from the crowd, many of whom were unaware the computers they collected and shipped away were causing so much suffering. Now, nearly all of the brand name computer manufacturers assiduously account for recycling done in their names. think that kind of primitive materials recovery is still going on, nonetheless, Grossman says. much of this equipment is coming from all over the world.

The Greening of IT goes beyond making less harmful hardware. Firms are looking to clean up their processe converse uk s and use of resources as well.

Wipro, for instance, tracks metrics such as carbon dioxide emissions per employee and per dollar of revenue, recycling versus landfill ratios and paper consumed per employee. Internet search engine giant Google is going a step beyond, having committed a energy czar to the task of finding a way to create one gigawatt or more of renewable electricity at a price that less expensive than producing it from burning coal. electricity use.

wish [Google] the best of luck, says Wharton White. lies a long way from Google core competency. They only make money one way and that selling ads.

Encouraging Consumers to Consume Less

Anal converse uk ysts, including Gartner, say that as environmental concerns come to be perceived as more pressing by consumers, retail customers will put increasing importance on indicators such as Energy Star labels and a company reputation as part of their purchase decision making.

Lexmark, the Lexington, Kentucky based maker of printers and toner, is encouraging its customers to print less, or at least to print smarter by using double sided printing. view what we doing as bringing a value to the retail customer, says Tanya Jackson, Lexmark director of sustainable development. have a focused effort on getting our customers to print less. In addition, the company has lowered its products energy consumption and launched a comprehensive program for reclaiming its used devices, breaking them down to their raw materials and using them to make new products.

converse uk regulated ternary complex with

regulated ternary complex with phospholipase C

Correspondence to: E C Kohn, 10 Center Drive, 10/2A33, Bethesda, MD 28092 1500, USACAIR 1/BAG 3 forms an EGF regulated ternary complex with Hsp70/Hsc70 and latent phospholipase C (PLC ). The expression of CAIR 1, CAI stressed 1, was induced in A2058 human melanoma cells by continuous exposure to CAI, an inhibitor of nonvoltage gated calcium influx. CAIR 1 sequence is identical, save 2 amino acids, to BAG 3 also cloned recently as Bis, a member of the bcl 2 associated athanogene family. We show that CAIR 1/BAG 3 binds to Hsp70/Hsc70 in intact cells and this binding is increased by short term exposure to CAI (Pin vivo in the absence of stimulation. Basal phosphorylation is inhibited by treatment with d erythrosphingosine (d ES), a broad inhibitor of the protein kinase C family. CAIR 1/BAG 3 contains several PXXP SH3 binding domains leading to the hypothesis that it is a partner protein of phospholipase C . PLC is bound to CAIR 1/BAG 3 in unstimulated cells. It is increased by CAI or d ES (P=0.05) treatment, and abrogated by EGF (r2=0.99); d ES treatment blocks the EGF mediated dissociation. We show that CAIR 1/BAG 3 binds to PLC and Hsp70/Hsc70 through separate and distinct domains. Hsp70/Hsc70 binds to the BAG domain of BAGs 1 and 3. CAIR 1/BAG 3 from control and EGF treated cell lysates bound selectively to the SH3 domain of PLC , but not its N SH2 or C SH2 domains. Confirming the SH3 interaction, PLC was pulled down by CAIR 1/BAG 3 PXXP GST fusions, but GST PXXP constructs confronted with lysates from EGF treated cells did not bind PLC as was seen in intact cells. Hsp70/Hsc70 was brought down by the PLC SH3 construct equally from native and EGF treated cells, but did not bind the PXXP construct under either condition. We propose that CAIR 1/BAG 3 may act as a multifunctional signaling protein linking the Hsp70/Hsc70 pathway with those necessary for activation of the EGF receptor tyrosine kinase signaling pathways. Intracellular calcium concentrations are tightly regulated and are altered during transmembrane signaling events, such as those stimulated by growth factors, and also in response to cellular injury or stress (Berridge et al., 1998; Bukau and Horwich, 1998; Zwick et al., 1999). We have identified an inhibitor of calcium influx in nonexcitable cells, CAI (Felder et al., 1991; Gusovsky et al., 1993; Kohn et al., 1994a). Exposure to CAI causes tumor and endothelial cell cytostasis and inhibits cell attachment, migration, and angiogenesis and tumorigenesis in vitro and in vivo (Kohn et al., 1992, 1994a,b, 1995; Kohn and Liotta, 1990). We hypothesized that continuous exposure to CAI by altering stimulated and basal intracellular calcium concentrations would stress the tumor cells. This would then either stimulate a stress recovery response or alter regulation or expression of proteins required for calcium activation pathways, such as the phospholipases. Activated phospholipases C produce inositol trisphosphate, a potent second messenger inducing internal calcium release and subsequent transmembrane calcium influx (Berridge et al., 1998). We developed a stress model in which tumor cell sublines were isolated after long term continuous exposure to increasing concentrations of CAI. We now report cloning of CAIR 1, CAI stressed 1, a protein nearly identical to BAG 3 (Takayama e converse uk t al., 1999) from CAI stressed cells, and demonstrate novel regulation and function of CAIR 1/BAG 3.

BAG 3 was cloned from a screen for homologs of the bcl 2 binding protein BAG 1 (bcl 2 associated athanogene) family (Takayama et al., 1995, 1999). BAG 1 was identified in a screen for bcl 2 binding proteins and has been shown to inhibit apoptosis and promote tumorigenesis (Stuart et al., 1998; Takayama et al., 1995, 1997). Recent homology cloning identified fragments of at least four other BAG family members, all of which contain a conserved C terminal domain, the BAG domain (Takayama et al., 1999). There is little other homology between BAG 1 and BAG 3, including lack of the BAG 1L nuclear localization signal or the BAG 1 ubiquitin region. The BAG domain was shown by pull down experiments and functional assays to bind to and promote substrate release from both HSP 70 and HSC 70 (Stuart et al., 1998; Takayama et al., 1997, 1999). Direct binding of BAG 1 to bcl 2 could not be demonstrated upon direct confrontation, however addition of ATP markedly enhanced BAG 1/bcl 2 binding, suggesting the requirement for an HSP 70 intermediate (Takayama et al., 1997). HSP 70 and its constitutive form, HSC 70, are molecular chaperones activated by varied cellular stresses including heat shock, oxygen free radicals, transition heavy metals, inflammation, ischemia, anti neoplastics, oncogenes, and proto oncogenes (Morimoto, 1998). They bind ATP through an ATPase domain and have a separate substrate binding domain (Bukau and Horwich, 1998; Pilon and Schekman, 1999). The site of Hsp70/Hsc70 binding to BAG 1 has been defined as the ATPase domain (Bukau and Horwich, 1998; Takayama et al., 1997); a 1 : 1 molar binding results in reduction in the refolding function of HSP 70 (Stuart et al., 1998). While other functions for BAG 1 such as binding to and activating Raf 1 have been reported, no link between those functions and HSP 70 have been demonstrated. CAIR 1/BAG 3 was recently cloned also as bis (Lee et al., 1999), in a protein interaction cloning procedure using bcl 2 as bait. Lee and coworkers demonstrated a weak anti apoptotic activity of transfected bis and further showed synergy when suboptimal doses of both bis and bcl 2 were transfected, simultaneously. They localized the site of int converse uk eraction of bis to the BH1 domain of bcl 2.

Phospholipase C (PLC ) regulates cytosolic free calcium concentration in response to transmembrane signal transduction, and has been linked to malignant transformation, invasive potential, and angiogenesis (Chang et al., 1997; Khoshyomn et al., 1999; Smith et al., 1998; Turner et al., 1996, 1997; Yang et al., 1998). Several investigators have demonstrated transforming capacity of both full length PLC and the isolated SH2 SH2 SH3 moiety (Bar Sagi et al., 1993; Chang et al., 1997; Schlessinger, 1994; Smith et al., 1998). Its link to calcium homeostasis, transmembrane signal transduction, and transforming potential made it a putative target protein in the calcium influx stress model. We demonstrate that CAIR 1, which is increased in expression in CAI treated cells, is a partner protein for PLC , and forms a ternary complex with HSP 70. This provides a previously unknown link between transmembrane signaling and the HSP 70 system.

Identification and cloning of CAIR 1/BAG 3

A2058 human melanoma cells were exposed chronically to continuing and escalating quantities of the calcium entry blocker, CAI. A subline previously shown to be sensitive in vitro and in vivo to the anti invasive and anti tumorigenic activity of CAI was used as the control for subtractive hybridization. A chemical cross link solution hybridization protocol was used as described to minimize loss of low copy number transcripts (Hampson et al., 1992) and yielded 35 non overlapping clones. A clone containing the 3′ 1200 bp was upregulated by approximately threefold and was chosen for further analysis. CAIR 1, CAI stressed 1, was cloned as a 2.6 kb transcript encoding a 74 kDa protein (Figure 1a), found by GenBank BLAST analysis to differ by 2 amino acids (227Q from a K; 237Q from an R) from BAG 3, a member of the bcl 2 associated athanogene family of bcl 2 binding proteins (Takayama et al., 1995, 1999). Genomic clones from chromosome 10q25 confirm the CAIR 1 sequence. Northern analysis revealed a twofold net increase in expression of CAIR 1/BAG 3 in cells stressed by constant exposure to 10 M CAI (10 Figure 1b) but not in cells exposed to 10 M CAI for 24 h (Figure 1c). Domain analysis of CAIR 1/BAG 3 reveals the previously reported BAG region (Takayama et al., 1999) and WW domain, as well as seven putative protein kinase C (PKC) and five putative CKII phosphorylation sites, multiple potential tyrosine phosphorylation sites, and a series of proline rich PXXP repeats of the SH3 binding type (Pawson and Scott, 1997). No genomic amplification of CAIR 1/BAG 3 gene in the CAI stressed cells was demonstrated by Southern analysis or fluorescent in situ hybridization (data not shown). Broad tissue expression of CAIR 1/BAG 3 was found in adult and embryonic tissues with lowest expression seen in hematopoietic tissues (Figure 1d). Genomic conservation was limited to the higher eukaryotes by zoo blot (not shown, Bios, New Haven, CT, USA). Immunoprecipitation from A2058 cells followed by immunoblot with anti peptide antibodies to CAIR 1/BAG 3 recognized a 74 kDa protein which was increased in quantity in 20 M CAI stressed cells (Figure 1e). No similar induction of CAIR 1/BAG 3 protein was observed in wild type A2058 or MDA 435 human breast cancer cells upon exposure to 10 M CAI for up to 24 h (Figure 1f), consistent with the lack of increased gene expression with acute CAI exposure.

CAIR 1/BAG 3 is phosphorylated in vivo

CAIR 1/BAG 3 has not been demonstrated previously to be a phosphoprotein. Domain analysis of CAIR 1/BAG 3 indicated the presence of potential PKC and CKII phosphorylation sites, as well as putative tyrosine phosphorylation sites. In vivo phosphorylation demonstrated that CAIR 1/BAG 3 is phosphorylated in unstimulated wild type A2058 cells and unstimulated MDA 435 human breast cancer cells (Figure 2). No change in in vivo phosphorylation of CAIR 1/BAG 3 was found after cell treatment with PMA (100 ng/ml) for 30 min to stimulate endogenous PKC or 24 h to down regulate PMA sensitive classical and novel PKC isotypes (Figure 2a) (Hata et al., 1993; Nishizuka, 1988). Incubation with a broad spectrum inhibitor of all classes of PKC isotypes, d ES (10 g/ml for 2 h), markedly reduced CAIR 1/BAG 3 incorporation of 32P o phosphate without effecting total cellular CAIR 1/BAG 3 (Figure 2b). No tyrosine phosphorylation of CAIR 1 was demonstrated in unstimulated A2058 or MDA 435 human breast cancer cells. EGF treatment of MDA 435 cells, which did not effect the quantity of CAIR 1, resulted in tyrosine phosphorylation of CAIR 1/BAG 3 (Figure 2c). CAIR 1/BAG 3 remained restricted to the cytosol upon EGF exposure. A small but statistically significant effect of CAI (10 M, 2 h) was seen on CAIR 1/BAG 3 basal phosphorylation in MDA 435 cells (87 of control, P of control, Pin vivo incorporation of 32P o phosphate could be detected after 1 min exposure to EGF (100 ng/ml; Pn=3; Figure 2d).

In vivo binding of CAIR 1/BAG 3 to Hsp70/Hsc70 is increased by CAI exposure

BAG 1 and BAG 3 have been shown through ex vivo binding experiments to bind to HSP 70 (BAG 1 only) and HSC 70 (both) through the newly described BAG domain (Takayama et al., 1999). We demonstrate in vivo binding of CAIR 1/BAG 3 to Hsp70/Hsc70 and further show its regulation by CAI treatment. An antibody recognizing both molecular forms HSP 70 and HSC 70 was used for the CAIR 1/BAG 3 studies. CAIR 1/BAG 3 co immunoprecipitated Hsp70/Hsc70 under unstimulated conditions from MDA 435 cells (Figure 3). Treatment with 10 M CAI for 90 min, a time that does not alter expression of either CAIR 1/BAG 3 or Hsp70/Hsc70, resulted in a 207% increase in bound Hsp70/Hsc70 to CAIR 1/BAG 3 (Pn=5). Nor did d ES exposure significantly change Hsp70/Hsc70 complex formation (Figure 3b).

CAIR 1/BAG 3 is a binding protein for latent PLC

PLC was chosen as a putative CAIR 1/BAG 3 binding protein because of its potential involvement in the response to CAI stress, because its activation has been shown to be CAI sensitive (Gusovsky et al., 1993), and because it contains an SH3 domain to which the CAIR 1/BAG 3 PXXP regions could target. Unstimulated A2058 and MDA 435 cells were used initially to determine if CAIR 1 interacted with PLC independently of growth factor receptor induced interactions. Untreated cells were lysed under high stringency conditions (Buffer A adjusted to 300 mM NaCl with added 0.1% Triton X 100 and 0.1% SDS) and subjected to immunoprecipitation with anti CAIR 1/BAG 3 Ab 2 or Ab 8. Immunocomplexes were detected by immunoblot for PLC (Figure 4a). Similar results were seen after precipitation with anti PLC antibody and blotting with Ab2 or Ab8; approximately 3 of total cellular PLC is bound to CAIR 1/BAG 3 under basal conditions (data not shown). No association of PLC with CAIR 1/BAG 3 was found by coimmunoprecipitation in MDA 435 cells under standard or high stringency RIPA lysate conditions (Figure 4c), indicating a selectivity of CAIR 1/BAG 3 for PLC .

EGF stimulation results in rapid diss converse uk ociation of PLC from CAIR 1/BAG 3

We hypothesized that phosphorylation of CAIR 1/BAG 3 would regulate its binding to PLC . EGF treatment of MDA 435 cells induced a dose dependent reduction of the binding of PLC to CAIR 1/BAG 3 (Figure 5a). The EGF dose response for this dissociation is log linear (r2=0.99) with an EC50 of 55 ng/ml. EGF induces a rapid loss of PLC binding from CAIR 1/BAG 3, with almost all loss of binding within the first minute (Figure 5b). The potential tyrosine phosphorylation sites in the BAG domain may regulate CAIR 1 Hsp70/Hsc70 complex formation. However, no significant effect of EGF treatment on this complex was observed (n=4, Figure 5c). Since d ES alters the basal phosphorylation status of CAIR 1/BAG 3, we tested its ability to alter binding to PLC . Cell treatment with d ES caused an approximately 150% increase in PLC binding to CAIR 1/BAG 3 in unstimulated cells (PP binding to CAIR 1/BAG 3 (P=0.49 control vs dES/EGF, Figure 6a). Treatment of MDA 435 cells with d ES did not effect the quantity of EGF receptor protein or the tyrosine phosphorylation status of the EGF receptor under the conditions used for the co immunoprecipitation studies. CAI (10 M) incubation for 90 min also increased binding of PLC to CAIR 1/BAG 3 without altering quantity of either protein (Pn=4, Figure 6b).

CAIR 1/BAG 3 binds to PLC and Hsp70/Hsc70 through separate and distinct domains

CAIR 1/BAG 3 binds to the PLC SH3 domain: PLC can interact with partner proteins through multiple domains, including its two SH2 domains and an SH3 domain (Koch et al., 1991; Pawson and Scott, 1997). CAIR 1/BAG 3 was pulled down from unstimulated MDA 435 cell lysates by the GST PLC SH3 domain but not t converse uk he PLC N SH2 or C SH2 domains (Figure 7a). Treatment with EGF of the cells donating CAIR 1/BAG 3 did not alter CAIR 1/BAG 3 binding to the PLC SH3 domain, in contrast to what was observed with whole cell coimmunoprecipitation studies where EGF treatment reduced binding of CAIR 1/BAG 3 to PLC in whole cell lysates (Figure 7b). No binding to SH2 domain constructs was seen after EGF stimulation. Hsp70/Hsc70 has been reported to bind to BAG 1 and BAG 3 through the BAG domain (Takayama et al., 1995, 1997, 1999). No whole cell in vivo co precipitation of PLC and Hsp70/Hsc70 was observed (data not shown). Reprobing of the PLC SH3 domain binding blot for Hsp70/Hsc70 demonstrated that the PLC SH3 construct pulled down Hsp70/Hsc70 from unstimulated and EGF treated cell lysates equally (Figure 7c).

PLC binds to the CAIR 1/BAG 3 PXXP domain: Pull down experiments were done using the PXXP4 region of CAIR 1/BAG 3, a region containing four different PXXP repeats (Figure 1) of SH3 binding type (Koch et al., 1991; Pawson and Scott, 1997). This region does not include the BAG domain. This was done to determine if this region is the PLC binding site on CAIR 1/BAG 3 and thus the potential mechanism through which PLC associates with Hsp70/Hsc70. A second construct containing the three C terminal PXXP repeats was also generated and tested. Both constructs bound PLC from unstimulated MDA 435 cell lysates (Figure 8a) demonstrating that the PLC binding site on CAIR 1/BAG 3 is a functional PXXP SH3 binding site. In contrast to the PLC SH3 experiments and consistent with the whole cell immunoprecipitation experiments, no or minimal binding of PLC from EGF treated lysates to the CAIR 1/BAG 3 PXXP4 domain was found. Further, no Hsp70/Hsc70 could be detected in the GST PXXP4 construct experiments (Figure 8b). These data indicate a novel role for CAIR 1/BAG 3 as a binding partner of latent PLC , that PLC binding to the CAIR 1/BAG 3 SH3 binding domain is regulated by EGF treatment, and that PLC and Hsp70/Hsc70 bind CAIR 1/BAG 3 through different domains.

We demonstrate that CAIR 1/BAG 3 forms a ternary complex between latent PLC and Hsp70/Hsc70. This is the first demonstration of a regulated crosstalk between Hsp70/Hsc70 and the EGFR PLC signaling pathway. The lack of binding of Hsp70/Hsc70 to the CAIR 1/BAG 3 PXXP PLC binding domain suggests that PLC and Hsp70/Hsc70 interact with CAIR 1/BAG 3 at distinct, non overlapping locations on the CAIR 1/BAG 3 molecule. Partner protein binding of both CAIR 1/BAG 3 with PLC and CAIR 1/BAG 3 with Hsp70/Hsc70 is increased by CAI, the nonvoltage gated calcium influx inhibitor used to stress cells for cloning of CAIR 1/BAG 3; d ES, the PKC inhibitor, increased CAIR 1/BAG 3 binding to PLC but not to Hsp70/Hsc70. Release of PLC from CAIR 1/BAG 3 is stimulated by EGF treatment but abrogated by exposure to d ES. CAIR 1/BAG 3 binds in vivo and in vitro to PLC from lysates from unstimulated cells but not from EGF treated cells whereas wild type PLC SH3 pulls CAIR 1/BAG 3 from both unstimulated and EGF treated cells. This suggests that the PLC SH3 domain is a target site in the EGF receptor signaling pathway and argues against tyrosine phosphorylation of CAIR 1/BAG 3 as playing a role in the release of PLC after EGF treatment. These data are consistent with the concept that CAIR 1/BAG 3 is a stimulus to bring Hsp70/Hsc70 into proximity of PLC to allow or facilitate the change in the PLC SH3 domain and subsequent release from CAIR 1/BAG 3 (Figure 9).

CAIR 1/BAG 3 is a protein containing multiple protein interaction motifs and phosphorylation sites (Figure 1), in addition to a conserved BAG domain (Takayama et al., 1999). We have focused on the PXXP region, a putative SH3 binding domain, and have demonstrated that this region binds the latent form of PLC . There is a rapid reduction in PLC binding to CAIR 1/BAG 3 seen after cellular EGF treatment. Release of PLC from CAIR 1/BAG 3 is rapid, nearly complete by 1 min. The kinetics of PLC phosphorylation as previously reported (Wahl et al., 1988; Yang et al., 1994, 1998) and confirmed by us in this model system shows the peak of phosphorylation occurring between 1 and 2 min. Maintenance of PLC binding is seen when a wild type PLC SH3 construct is used but a marked reduction in partner protein complex is observed when the CAIR 1/BAG 3 PXXP region is presented with lysates from EGF treated cells. This implicates a change(s) in the conformation of the PLC SH3 domain is necessary for the process of CAIR 1/BAG 3 release of PLC upon EGF stimulation.

The BAG domain of CAIR 1/BAG 3 contains two tyrosine and three threonine residues that are not conserved from the BAG domain of BAG 1 (Takayama et al., 1999). We demonstrate endogenous binding of CAIR 1/BAG 3 to Hsp70/Hsc70 and show an increase in complex formation when cells are stressed for 90 min by exposure to 10 M CAI. Previous studies have shown a BAG 3 interaction with Hsp70/Hsc70 using BAG GST pull down experiments, from BAG 3 expressed in intact cells, and using a kDa recombinant BAG 3 fragment (Takayama et al., 1999). These studies demonstrated that the 24 kDa BAG 3 fragment had an attenuated binding to Hsc70 in a BIA Core assay and that Hsc70 dissociated more rapidly from the BAG 3 fragment BIA Core chip than did the purified BAG 1 or BAG 2 molecules. Our demonstration of endogenous binding of CAIR 1/BAG 3 to Hsp70/Hsc70 confirms that the interaction of CAIR 1/BAG 3 occurs in intact cells. The increase seen after

converse uk Regular Hampton contributor of

Regular Hampton contributor offers savings ideas for kitchen treats and bathroom cleaning

One of the joys of winter is the variety of citrus available in local stores. One of the mysteries is why clementines alone come packed in real wooden crates as opposed to the pedestrian string or even plastic bags that typically encase the season’s oranges and grapefruit.

It’s a good question. What do you do with those attractive crates? Kelly hates to throw them away, so please help her out.

while we’re asking questions, does anyone have any suggestions for what to do with pantyhose that’s no longer presentable? One recycling use is to cut them up and use them for stuffing pillows. Other than that we’re stymied and unless you’re making a lot of pillows they can pile up pretty quickly better still would be if you have a suggestion for preventing runs on the first or second wearing. That would be a real savings for any woman whose job’s dress code requires they wear converse uk hose.

question. Some grocery store converse uk s offer a nickel discount for every plastic bag you don’t use of theirs when you bring in your own shopping bags. Let us know if your local store offers this saving and also, which recyclable bags you find work the best.

Kitchen and bathroom savingsBarb Cathcart of Hampton, the “queen of thrift,” is never short of ideas for how to save a nickel or two.

Here are a couple more of her suggestions for how to save in the kitchen and on cleaning materials.

To clean tile: “I take a clean squirt bottle and add a few tablespoons of bleach, then fill the remaining area with water. Every time you take a shower or bath, squirt the tile no mold or mildew . ever!” Cathcart reports.

She issues a couple of cautions though: Be sure to mark the outside of the bottle so that it’s not mistaken for plain water for misting plants or anything else. Also, be sure to exit the bathroom after spraying converse uk due to the fumes. Turn on the fan and shut the door behind you for a few minutes. To make Cathcart style “doughnuts”: Take a can of 10 Pillsbury Biscuits or similar, pop open and separate the biscuits. In a shallow frying pan, place about 1 to 2 inches of your favorite oil (canola, safflower, etc.) and heat. Make a hole in the center of each biscuit, drop into hot oil, and be ready to flip in less than a minute. Each side usually takes less than a minute depending on the temperature of your stove. Drain the “doughnuts” on a paper towel and coat with your favorite topping, such as canned frosting, sugar, cinnamon, etc. Yum (and probably no more c converse uk alories than a regular doughnut).

converse uk regret dole commentsKen O’

regret dole comments

Ken O’Dowd reportedly told a community forum in his central Queensland electorate of Flynn this week: “You won’t get anyone on the dole coming to these sort of meetings, because they don’t care about the community, they care about themselves and how they can screw the system”.

The MP’s remarks were made shor converse uk tly before the announcement of a government review of welfare payments.

Mr O’Dowd also told the forum about a recent conversation he had with billionaire mining magnate Gina Rinehart, in which she voiced her concerns about the welfare system.

She told him that 60 per cent of Australians were on some sort of welfare payment, questioning whether they were all that “badly off”.

Mr O’Dowd wa converse uk s not taking a backward step on Thursday as his remarks received wider converse uk coverage than a local newspaper.

But he insisted his accusation was aimed at long term unemployed people rec converse uk eiving the disability support pension who “deliberately try to be on the dole”.

converse uk Registering units a must for s

Registering units a must for safety

That the message from Newmarket bylaw officers who have been visiting, speaking and leaving information with residents about changes to the rules for basement apartments, in law suites and secondary suites that are included under the ADU umbrella.

In the spring, council approved changes that require all homeowners with an ADU to register their units through a one time applic converse uk ation process and fee of $250.

your ADU is first and foremost a matter of health and safety, notes Mayor Tony Van Bynen. a critical situation, responders need to know where the emergency is, who is affected and if everyone is accounted for.

These new rules will help with that, he says.

If you plan to rent, watch for regi converse uk stered units that will be identified with an plate on the front of the home as well as a address that will help emergency responders know there are two units in the home, which could save time and, potentially, lives.

Newmarket resident John Dowson app converse uk lauded town staff for drafting such a detailed bylaw governing rules around home apartments after consulting the public, but wanted to voice other major concerns.

most common complaint about ADUs is the garbage, Mr. Dowson told councillors when speaking on behalf of his neighbourhood association.

According to the recent bylaw, houses with apartments are allowed six bags of garbage compared to a single family occupied home, which is allowed three. These bags are often stored at the front of the house where animals and the weather have the opportunity to spread the garbage around the street. Mr. Dowson asked council to consider stipulating garbage must be stored in a container.

Parking is another common problem with tenants parking on lawns or out in the street because units aren required to widen their driveways to provide the required four parking spaces for ADUs.

Finally, he asked that council educate not only the property owners but also the occupants about the bylaw by providing a package or something similar. He also suggested harsher punishment for infractions.

consequences there will be no compliance, he added.

Town clerk Andrew Brouwer informed Mr. Dowson the education aspect can be included converse uk in the communication strategy for the bylaw.

Bylaw services supervisor Lesley Long informed Mr. Dowson when the bylaw states garbage must be kept in a container, a garbage bag is included in the definition of a container. but if the garbage is strewn across the neighbourhood, enforcement of the property standards bylaw would cover the issue.

She also explained it wouldn be possible to include a requirement that property owners with ADUs require a driveway widening as zoning bylaws restrict the percentage of a property that can be driveway, meaning some properties wouldn be able to widen to accommodate four spaces.

long as there are four spaces available on the exterior (not including garage space) it is not an issue, she said. going forward, the town may be able to issue tickets to those parking on lawns and use property standards to encourage the property owner to address the problem.

The bylaw also sets ADU fees of $10 for a replacement plate fee and $100 for a refusal and revocation appeal fee.

Council also added that registrations of ADUs will be revoked following three bylaw strikes under the property standards, zoning, noise and garbage bylaws over five consecutive years at one address.

converse uk Registered sex offender arrest

Registered sex offender arrested after Ohio police find three bodies wrapped in plastic bags

East Cleveland Police Chief Ralph Spotts had cautioned searchers to be prepared to find one or two more victims. But after a daylong search that included 40 abandoned houses and other areas, no more bodies were found.Mayor Gary Norton said the suspect has indicated he might have been influenced by Cleveland serial killer Anthony Sowell, who was convicted in 2011 of murdering 11 women and sentenced to death.It the latest in a series of high profile cases involving the disappearance of women from the Cleveland area.The odor led to the discovery Friday of one body in a garage. Two others were found Saturday one in a backyard and the other in the basement of a vacant house.Searchers rummaging through vacant houses in the same neighborhood Sunday were warned by Spotts to brace themselves for the smell of rotting bodies and to look out for trash bags that might conceal a body.Spotts indicated later Sunday that the suspect comments haven provided clarity on whether more bodies might be found.really hasn stated that there any more, but he hasn said anything that would make us think that there not, Spotts said.said some things that led us to believe that in some way, shape, or form, Sowell might be an influence, Norton told The Associated Press.It wasn immediately clear whether Madison has an attorney, and no one was commenting Sunday afternoon at the address his mother home.Madison was classified as a sex offender in 2002 when he was sentenced to four years in prison for attempted rape, according to Cuyahoga County court records. He had previous convictions in 2000 and 2001 for drug related charges.All three bodies were found in the fetal position, wrapped in several layers of trash bags, Norton said. He said detectives continue to interview Madison, who used his mother address in Cleveland in registering as a sex offender, the mayor said.Cuyahoga County medical examiner Dr. Thomas P. Gilson said Sunday that the bodies were in advanced stages of decomposition and th converse uk at it would take several days to identify them and how they died.About three dozen volunteers, including community anti crime activists, fanned out Sunday morning converse uk across yards, through vacant houses and along a railroad to help police search. The chief advised them to watch for missing floor boards as they looked inside houses. One young searcher crawled under a board screwed across a door to go inside a house to search.MO of each body we found so far was wrapped up in a lot of garbage bags, so if you see anything and it might not look like it a body, but it could be because each bag, the way he had each person was in a fetal position, Spotts told searchers before they began. didn look like a person could actually fit in the bag. Butcher, 55, said she came out to help search her neighborhood because she was disturbed by the death and said she knew other volunteers were, too.are concerned because it could have been one of their family members, she said. could have been one of their kids. It could have been one of their nieces. It could have been one of their aunts. neighbor, Nathenia Crosby, said she was familiar with Madison and had seen him walking through the neighborhood. She said she had told him to stop chatting with her daughter and warned him after seeing him talk converse uk to her cousin.very scary, especially when he used to be talking to my daughter, said Crosby, 48. I told him he was too old to be talking to my daughter because she was only 19. When I found out how old he was, I said, You need to move on, she too young. day earlier, police, FBI, the Ohio Bureau of Criminal Investigation and the Cuyahoga County Sheriff Department went through yards and abandoned houses over about three blocks and used dogs trained to find cadavers.we pray to God, this is it, he said.The case brings to mind recent notorious Cleveland searches that involved missing women.In May, three women who separately vanished a decade ago were found captive in a run down house. Ariel Castro, a former school bus driver, has pleaded not guilty to nearly 1,000 counts of kidnap, rape and other crimes.In 2009, Sowell was arrested after a woman escaped from his house and said she had been raped there. Police found the mostly nude bodies of 11 women in garbage bags and plastic sheets throughout the home.Prosecutors described him in court papers as worst offender converse uk in the history of Cuyahoga County and arguably the State of Ohio. was found guilty in 2011 and sentenced to death.

converse uk Register for an internship

Register for an internship

There are three basic steps to register for an internship: find an internship, find a faculty advisor and complete the necessary paperwork.

NOTE: At the latest, students considering pursuing internships should prepare during the quarter preceding the one in which they plan to register for the internship. For example, if you plan to register for an internship during the spring quarter, you should start looking for a site in the winter quarter. This means the BEGINNING of that quarter; not the end of the quarter. Be aware that many internship applications have deadlines; especially the ones for government agencies a converse uk nd larger corporations. You need to start looking early. Please remember that you need to register for your internship by the 10th day of the quarter in which you plan to participate in the internship.

Jim Stephenson, an Environmental Sciences student, spent his summer in an internship at Manchester State Park. Jim researched shoreline erosion by collecti converse uk ng pre restoration data on the intertidal invertebrate assemblage and using GIS to characterize and delineate habitat type.

Find an internship site:

Determine the type of work you want to do and identify the learning objectives you want to accomplish by engaging in this type of work.

Select the work site and identify a site supervisor. You may identify your own potential work sites within the community or make an appointment to discuss possible sites with the IAS internship coordinator. Local businesses, government, non profit agencies and educational institutions regularly utilize interns.

Some IAS faculty members know community members who would be excellent site supervisors and can help you make this connection. Contact a supervisor at the potential work site to discuss an internship for you.

Find a faculty sponsor:

Make your request once you have found an internship.

The faculty member supervises the academic component of the internship. Faculty sponsorship of internships is at the discretion of the IAS faculty.

A natural fit is a faculty member who teaches courses that match the content of your internship.

It’s best to approach a faculty member with whom yo converse uk u already have a working relationship.

Determine the number of credits you wish to attempt. Generally, three (3) hours/week on site over the course of an 11 week quarter will earn one (1) credit. Check with an IAS advisor or faculty member about appropriate placement of these internship credits in your program of study. Remember, internships are offered only on a credit/no credit basis.

Complete the Paperwork:

Once you, the faculty member and the site supervisor have an agreed upon plan, complete the Internship Contract Form. Forms must be typed. It is your responsibility to obtain the appr converse uk opriate course number and SLN from the quarterly registration guide. Sign the completed form and make copies of it (the original will be kept in the IAS program office) so you have copies for your faculty sponsor, your site supervisor and for your own records. Submit it to the IAS internship coordinator for review and signature before being issued an entry code. Once you have received an entry code, you are allowed to officially register for the internship.

converse uk Regional ServiceThe amount

Regional Service

The amount of the contract is designed to cover ETS operating costs for the service. Please note the following requirements:

The C Line is operated by the City of Leduc. Edmonton Transit fare products are not valid on C Line service, but may be used once you arrive in Edmonton; C Line passes are only valid on C Line buses, not on ETS service. As a service to transit customers, both ETS and C Line bus passes are available for purchase in Leduc.

Post secondary students with a valid U Pass ride for $1 for a one way fare

All passenger luggage must be placed on the seat or on the floor in front of the seat. No passenger luggage will be allowed in the aisle.

Provincial requirements state that the maximum standing load on a converse uk bus is 25 standees.

First come / first served process is in operation for the service.

Edmonton Transit is not responsible for lost passenger luggage. Call 780 496 1622 for Lost/Found.passenger luggage includes: suitcases, backpacks, boxes, bags and any other carrying device for the transport of personal belongings.

St. Albert Transit Service

The City of St. Albert operates local bus service within the City of St. Albert and commuter bus service to and from various locations within the City of Edmonton. Service operates Monday to Sunday.

Effective February 1, 2013

St. Albert Transit Fares

When travelling on ETS service: St. Albert “Commuter” passes, tickets and valid transfers are accepted on ETS service (St. Albert tickets are not valid for LRT travel). St. Albert “Local” passes, tickets and transfers are not valid on ETS service.

When travelling within Edmonton: All ETS fare products are valid on St. Albert service travelling within Edmonton.

When travelling locally within St. Albert: All ETS fare product, except the Seniors Monthly Pass and the Courtesy Pass, may be used with the payment of a $1 surcharge. (Note: ETS DATS Passes are not valid on St. day passes, monthly passes, annual pa converse uk converse uk sses) except the Seniors Monthly Pass and the Courtesy Pass, may be used for cross boundary travel on St. Albert service with the payment of a $1 surcharge.

ETS tickets and valid transfers may be used for cross boundary travel on St. Albert service with the payment of a $2 surcharge.

For route and schedule information, please visit the

St. Albert Transit website or call St. Albert Transit Customer Services Office at 780 418 6060.

Strathcona County (Sherwood Park) Transit Service

Strathcona County operates local bus service within Sherwood Park and regional bus service within Strathcona County and the City of Edmonton. Service operates Monday to Sunday.

Effective February 1, 2013

Strathcona County Transit Fares

When travelling on ETS service: Strathcona “Commuter” passes, ticke converse uk ts and valid transfers are accepted on ETS bus service (tickets are valid for bus service only, not for LRT travel). Strathcona “Local” passes, tickets and transfers are not valid on ETS service.

When travelling within Edmonton: All ETS fare products are valid on Strathcona service travelling within Edmonton.

converse uk Regional roundSreenivas ba

Regional round

Sreenivas bags title

Host CIAL Golf Club’s Sreenivas won the Rotary Cup golf tournament for amateurs here on Sunday. He won the ‘best golfer’ title in the gold category with 42 points. Anil Menon, also from the same club, won the silver category.

1. Sreenivas (CIAL G 2. Rajeev Kumar (NEPTA Club);Anil Menon (CIAL), 2. Alexander (NEPTA, maximum birdies, 2).

Viveka upsets Aditi

Unseeded Viveka Sivaraman (TN) upset second seeded Aditi Narayan (TN) in the quarterfinals of the AITA under 12 Talent Series tennis tournament here converse uk on Sunday.

Viveka scored a crushing 4 0, 4 1 win over Aditi. Fourth seeded Netra Krishna of Ker converse uk ala lost to unseeded Kaavya Palani of Tamil Nadu in the quarterfinals.

In the boys’ section, fourth seeded converse uk Sahil Savasree (Karnataka) was beaten by Yognath Sudrith (TN) 5 3, 4 2 in the pre quarterfinals. Sabarinath (Ker) 4 0, 4 0; Hithen Sreelesh (Ker) bt Sam Prakash D. Jiffry 4 2, 4 0; Yognath Sudrith (TN) bt Sahil Savasre (Kar) 5 3, 4 2; M. In the quarterfinals, Customs beat Marian College of Engineering, Thiruvananthapuram 22 15; KSEB Whites beat SB College, Changanassery ‘A’ 11 8; KSEB Blue crushed Kerala Police ‘B’ 22 7 and Kerala Police ‘A’ beat SKV College, Thrissur 11 6.

In the semifinals, Customs will meet KSEB White while KSEB Blue will take on Kerala Police ‘A’.

South Zone ‘B’ in lead

South Zone ‘B’ took the first innings lead against Central Zone ‘B’ on a bizarre first day that saw 24 wickets fall at the KCA stadium here on S converse uk unday in the inter zone under 19 cricket tournament.

But Central Zone, in the second innings, found itself in a spot of bother again at 65 for four at stumps.