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Marinetrans and BGL have sold a majority stake to Waterland Private Equity
The shareholders of Best Global Logistics (BGL) and Marinetrans, both logistics service providers, have completed a combined sale of the companies to Waterland Private Equity Investments (Waterland).
Marinetrans is the world's largest fourth-party logistics service provider dealing exclusively with marine spare parts forwarding and logistics. The company is headquartered in Singapore and has six additional offices around the globe. Through a network of dedicated partners, Marinetrans uses 22 warehouses and provides services in 2,000 ports worldwide.
BGL is an independent third-party logistics service provider offering relatively complex logistics solutions on a global scale. The company operates two bonded and free-zone warehouses located in Lijnden and Rotterdam, the Netherlands. BGL has a strong foothold in the maritime industry, while also serving a range of other industries.
Founded in 1999 and headquartered in Bussum, the Netherlands, Waterland is an independent private equity investment group with eight offices across Europe. The company acts as an active shareholder and aims to play a key role in strategic and operational development, growth and performance.
Oaklins' team in the Netherlands advised the sellers in this transaction.
Michel Loots
Co-CEO and shareholder, Best Global Logistics
Talk to the deal team
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It hardly pays to complain about a poor patient who doesn't understand what it takes to get an off-formulary item, like a citrate solution for him, to treat his renal tubular acidosis, when he can't understand why he should take the awful tasting stuff, and my Spanish, even with the ministrations of Miss Coesfeld at RHS, back in Chicago, leaves me short of palabras. This poor patient, not qualifying for MediCal (Medicaid in the other 49 states, a combination of federal and state funding for the medically needy) for reasons only known to the bureaucrats.... the folks whose job it is to enlarge the holes in the "safety net," .... the poor guy, not feeling very well after running out of his medication, and perhaps thinking that he already used up the bottle, so now he should be well, was instructed to go to a hospital where the doctor who covers for me is on staff.... so when I leave for Hawaii, there will be a nephrologist there to frustrate him in my absence. No one to take him to the hospital and there's a hospiital around the corner, so that ought to do, in his mind. So, who's going to look after him when I'm continuing my medical education? That could be the subject of another boring 500 words and result in expelling me to a medical Devil's Island. Remind me to tell you about Medical Services to the Indigent, or MSI, sometime.... well, I'll handle it.
Tom Jones, that great and sexy (at least to the ladies, and to my wife, who once took me with her to sit in the very first row, dodging panties and keys) singer of Delilah, and many others, was to accompany us to the beach,
first, as a surprise, to the lovely Lin, and, when I couldn't hold back, and told her about the i-Pod and stuff, as kind of a promise. He's coming, but probably as a CD. Somehow, I have to learn how to get him from CD to i-Pod, and I'm of a generation that felt challenged by Tinker Toys, Erector Sets and balsa and tissue paper model airplanes. Of course, son, Josh, will be there for the second week, and his job is to keep Boeing's satellites spinning in orbit and delivering appropriate electronic information to the distant mortals... a cinch for him!
Enough of priorities, Hawaii and Tom Jones. March Madness. We, Americans, if we are basketball fans, understand this laser sharp focus on our raison d'etre (That's for Miss Coesfeld, and June, if she ever gets here.)
The remainder is basketball, and Bruins specifically. Others may tune out. What the heck is my son, Dirk, doing these days? He's a fan of Arizona (Wildcats).... at least they're in the Pac 10! He also loves Syracuse (New York)... the Orangemen! Arizona plays California (home of the Bears, Rhea.... not the Bruins!) WE, are the BRUINS.... UCLA! Fight, fight, fight! Bruins understand this without further comment.
This is the first day of the Pac 10 tournament and at 2:50, FSNW will have the Bruin-Beaver (Oregon State) game.
The OC Register says the Bruins are a 5 point favorite. I make it out to be 3. I'll settle for a 1 point win. I think, should that major accomplishment occur, the Bruins will play the Wildcats, and, if past performances mean anything, lose, and lose badly. Of course, sports don't always follow the odds and every time you play, theoretically, you can win. I had a tennis career like that. I could never beat a guy named Mel, and was relegated to playing doubles as he was the guy I had to beat to play singles for RHS. I always had a chance, and I always lost, well, not quite, but that's another story.
Today, the Bruins must find an answer for David Lucas, whose father, Maurice, was an NBA star. We have Twin Towers, Fey and Hollins, and they each have strengths, but if we used 6 guys, Fey and Hollins to cover Lucas, we still couldn't handle him! He's good. Instead, Coach Howland will use one of the 2 and Dijon Thompson, a very fine forward, slightly miscast as a "strong" forward. Dijon is all Pac 10, as is Lucas, but Lucas is muscle and Dijon is finesse and speed, and that is the challenge! Of course, when you double team, you leave someone open, and Lucas has spent his life being double teamed so he's looking for the pass off to the guy who is open, and there's where team defense comes into play. Switching is essential. It's that easy. So, in theory, was beating Mel. He wasn't that tall, he just ran down every ball I hit and out-steadied me into tennis oblivion. Of course, though he is now a retired pediatrician in Fremont, CA, he won't be at the Mauna Kea, and now, at age 69, I'm nearly ready for doubles.
Hope I can post from Paradise!

subset: commoncrawl


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Wales Go Yellow and Black Tonight!
5 September 2017, 16:50 | Updated: 5 September 2017, 17:57
Wales have unveiled a one-off shirt for the crucial game against Moldova.
Wales have recently worn a grey kit for away matches. Considered unlucky by some supporters, Chris Coleman's boys have yet to pick up a win whilst wearing it.
It won't be the first time Wales have worn yellow, with a number of different striking numbers through the years.
We're ready! Ydych chi'n barod hefyd? #MDAWAL #TogetherStronger pic.twitter.com/tWBF4jKd1B— Wales (@FAWales) September 5, 2017
Many fans have taken to it already, comparing the black and yellow colour scheme to the Flag of Saint David - a black and yellow cross.
Unfortunately though, for those hoping to get their hands on their own, the FAW have already confirmed there are currently no plans for it to go on sale.
The yellow away shirt for tonight's Wales match is #Wales— John Andrew Jones (@JohnAndrewJones) September 5, 2017
The new yellow & black #Wales away shirt is lovely. Shame it's only a one-off for tonight. Hope enough fans request it to make it permanent.— Mark Carter (@MarkLewisCarter) September 5, 2017
Wales Yellow kit for tonight is unbelievably lush #comeonwales #TogetherStronger #WalesAway— Lucas (@lukestamford) September 5, 2017
Loving that yellow Wales jersey— Keenan Watkins (@keenanwatkins) September 5, 2017


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Meghan Markle birthday: Why will Meghan NOT be allowed to keep all her birthday gifts?
MEGHAN Markle, Duchess of Sussex, will be celebrating her first birthday as part of the royal family today, but things are expected to be very different for her this year. Why can't she keep all of her gifts?
The Duchess of Sussex will be celebrating her 37th birthday today as the Royal family's newest member.
Since marrying husband Prince Harry, Meghan has enjoyed number of public outings and events as a working royal.
While there are many perks to being a part of the Royal family, there are strict traditions Meghan must adhere to.
Some of these surround conduct during birthdays, which could mean Meghan has to forfeit some of her presents.
Why will Meghan have to return presents?
Royal tradition means that the royals aren't allowed to host huge lavish parties, instead settling for a small family affair.
Certain unsolicited gifts will have to be turned away as presents from celebrities, businesses and firms are forbidden.
This is to prevent royals being exploited for commercial gain by companies and is a hardline rule in the house of Windsor.
Meghan Markle birthday: Why won't she get to keep some of her gifts?
This year, when Harry and Meghan were married, the couple were forced to return a total £7million worth of presents.
Official guidelines read: "Gifts offered by private individuals living in the UK not personally known to the Member of the Royal Family should be refused where there are concerns about the propriety or motives of the donor or the gift itself."
This year, the Duchess of Sussex has already been sent an expensive £290 handbag designed by Alexandra K from American animal rights organisation PETA.
Meghan will likely have to return this item alongside any others in order to keep an impartial image as a royal.
What will Meghan do for her birthday?
Megan's birthday this year could see some huge gifts coming towards her from the royal family.
Last year, it was rumoured that Prince Harry got the Duchess a painting he hand-selected from a Kensington gallery.
The Prince also apparently took his then-girlfriend out for a trip to Botswana.
This year Meghan will be at the wedding of Charlie van Straubenzee on her birthday.
A close friend of both William and Harry, Van Straubenzee was also present at Harry and Megan's wedding in May.
The couple will be joined by Prince William, who will likely bring Kate with him, making Meghan's birthday a social and family-led occasion.
Otherwise, very little is known of the plans for the day, but Meghan has said in the past her mother has told her to treat the day like "your own personal New Year".


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In the development of a component vaccine against caries, the catalytic region (CAT) and glucan-binding domain (GBD) of glucosyltransferase B (GtfB) from Streptococcus mutans have been employed as target antigens. These regions were adopted as primary targets because they theoretically include epitopes associated with enzyme function. However, their antigenicities have not been fully evaluated. Although there are many reports about successful vaccination using these components, the principle has not yet been put to practical use. For these reasons, we came to doubt the effectiveness of the epitopes in vaccine production and reevaluated the antigenic region of GtfB by using in silico analyses combined with in vitro and in vivo experiments. The results suggested that the ca. 360-amino-acid variable region (VR) in the N terminus of GtfB is more reactive than CAT and GBD. This region is S. mutans and/or GtfB specific, nonconserved among other streptococcal Gtfs, and of unknown function. Immunization using an adenovirus vector-borne DNA vaccine confirmed that VR is an epitope that shows promise for the development of a caries vaccine.
Despite some success with preventive measures, dental caries continue to be prevalent and costly in many populations, and the development of a vaccine to prevent dental caries is still a desirable goal. Attempts to develop a vaccine against dental caries began with immunization using whole cells of the primary etiologic agent, Streptococcus mutans, in rhesus monkeys (23, 24). Although dental caries were successfully prevented in these studies, it was reported that the induced antibodies showed cross-reactivity with the human heart muscle (10, 48). Thereafter, components derived from S. mutans were investigated to avoid induction of antibodies that cross-reacted with human tissue. The molecules primarily focused on as target antigens are glucosyltransferase B (GtfB) (46) and antigen I/II (Ag I/II) (34).
GtfB was further truncated, and the catalytic region (CAT) (37) and the glucan-binding domain (GBD) (20) were employed as effective antigenic components. These components were combined with the cholera toxin B subunit (22), the saliva-binding region derived from Ag I/II (50), or both (30) to further enhance immunogenicity.
Despite these successful results reported more than 10 years ago, a GtfB-based vaccine against caries has not yet been put to practical use. One reason may be that induction of mucosal immunity is needed to produce the effective secretory IgA (S-IgA) antibodies that can prevent dental caries. Although encouraging results have been reported (3–6, 30, 38, 39), salivary S-IgA responses are often variable, transient, and of low magnitude compared with serum IgG responses. Another cause for the delay of the anticaries vaccine may be the antigenicity of the target antigens CAT and GBD, which were selected due to their association with enzyme function (40, 41). No study has addressed the immunogenicity of CAT and GBD by immunological, biochemical, or bioinformatic methods. It was recently suggested that effective screening for candidate vaccine antigens should include in silico analysis followed by in vitro and in vivo evaluations of antigenicity to reduce time and cost (29).
In this study, we reevaluated the immunogenicity of GtfB by using in silico approaches followed by in vitro and in vivo experiments to identify antigenic regions that may be employed in an effective anticaries vaccine. On the basis of our findings, we constructed a DNA vaccine using an adenovirus vector and confirmed its ability to induce specific antibodies contributing to immunity in mice.
Bacterial strains and growth media.S. mutans MT8148 (serotype c) was routinely cultured in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, MI) or on Mitis-Salivarius (MS) agar (Difco) at 37°C in air. Escherichia coli XL-2 (Stratagene Ltd., Cambridge, United Kingdom) was cultured aerobically in Luria-Bertani (LB) medium. When appropriate, erythromycin, kanamycin, and ampicillin (Wako Pure Chemicals, Osaka, Japan) were added to the LB medium to final concentrations of 500, 30, and 100 μg/ml, respectively.
Preparation of GtfB.S. mutans MT8148 was cultured in 5 liters of TTY medium (composed of Trypticase, tryptose, yeast extract, salts, and 1% glucose) instead of BHI broth, because it has been reported to produce an increased yield of bacterial cells (16) at 37°C, and grown to an optical density of 0.8 at 550 nm. The bacteria were collected by centrifugation, and cell-associated Gtf (CA-Gtf) was extracted by treatment with 50 ml of 8 M urea at 25°C for 1 h. The extract was dialyzed against 10 mM sodium phosphate buffer (NaPB; pH 6.5) and adjusted to 60% saturation with ammonium sulfate. After centrifugation, the precipitate was dissolved in 10 mM NaPB (pH 7.5) and then dialyzed against the same buffer. The crude CA-Gtf sample was applied to a DEAE-Sepharose FF column (bed volume, 10 ml; GE Healthcare UK Ltd., Buckinghamshire, United Kingdom) and eluted with a linear gradient of 0 to 1.0 M NaCl in the same buffer. Active fractions, measured as described below, were pooled, concentrated by ammonium sulfate precipitation, dialyzed against 10 mM NaPB (pH 7.5), applied to a Bio-Scale CHT10-I column (bed volume, 10 ml; Bio-Rad Laboratories, Hercules, CA), and then eluted with a linear gradient of 10 to 500 mM potassium phosphate buffer (KPB) (15). To select the GtfB fractions, we performed a glucan synthesis assay (15), an enzyme-linked immunosorbent assay (ELISA) using anti-CA-Gtf antibody, and a Western blot assay using anti-Gtf-I (GtfB) and anti-Gtf-SI (GtfC) monoclonal antibodies (47).
Glucan synthesis assay.Glucan-synthesizing activity was determined using [14C]glucose-sucrose as described previously (15). In brief, reaction mixtures composed of Gtf and 10 mM [14C]glucose-sucrose (11.47 GBq/mmol) in 20 μl of 50 mM KPB (pH 6.0) were incubated for 1 h at 37°C, spotted on a filter paper (1.0 by 1.5 cm), and air dried. The filters were washed with methanol to remove nonincorporated [14C]glucose-sucrose and then immersed in scintillation fluid to estimate the amount of de novo-synthesized [14C]glucan.
Generation of anti-CA-Gtf antiserum.Antisera were prepared by repeatedly injecting rabbits intramuscularly with the emulsion made by mixing Gtf-active fractions that were purified using a DEAE-Sepharose FF column and Freund's complete adjuvant (Difco) once, followed by Freund's incomplete adjuvant (Difco) twice. The antibodies were purified from rabbit antiserum by 3× precipitation with 33% ammonium sulfate.
SDS-PAGE and Western blotting.Conventional SDS-PAGE and Western blot analyses were carried out as described previously (12). In brief, Gtf samples and E. coli cells carrying the recombinant plasmid were suspended in SDS gel loading buffer and boiled for 5 min. Proteins separated by SDS-PAGE were transferred onto a polyvinylidene fluoride (PVDF) membrane (Immobilon; Millipore, Billerica, MA). After blocking with 5% bovine serum albumin (BSA), the membrane was reacted with a primary antibody at 37°C for 1 h, and the bound antibody was subsequently detected using a solid-phase immunoassay employing goat anti-rabbit IgG antibody conjugated with alkaline phosphatase (AP) or goat anti-mouse IgG antibody conjugated with horseradish peroxidase (HRP).
When NuPAGE 4-to-12% bis-Tris gels (Invitrogen by Life Technologies, Carlsbad, CA) were used, the samples were suspended in NuPAGE LDS sample buffer and NuPAGE sample reducing agent (Invitrogen) and incubated at 70°C for 10 min. Coomassie brilliant blue (CBB) staining of the gels was performed using SimplyBlue SafeStain (Invitrogen). For immunoblotting, the proteins were transferred onto 0.2-μm PVDF membranes (Invitrogen) for 30 min at a constant voltage of 200 V. After blocking with 5% nonfat dry milk and 0.2% Tween 20 in Tris-borate-saline (TBS) at 4°C overnight, the membranes were incubated with primary antibody in TBS containing 1% BSA at room temperature for 1 h. The membranes were washed 5 times with TBS containing 0.2% Tween and then incubated with secondary antibodies conjugated with HRP at a dilution of 1:3,000 in TBS with 1% BSA at room temperature for 1 h. The membranes were then washed 5 times with TBS, and the signals were detected using a conventional solid-phase immunoassay.
ELISA.The ELISA was used to measure the amounts of GtfB or titers of antibody. The GtfB samples were applied to a 96-well plate, incubated at 37°C for 1 h, and washed 5 times with phosphate-buffered saline (PBS) containing 0.2% Tween 20 (PBST). The primary antibody at a dilution of 1:1,000 in PBST was applied onto each well, and the plates were incubated at 37°C for 1 h and then washed 5 times. The appropriate secondary antibody conjugated with AP at a dilution of 1:1,000 was applied to each well and allowed to react at 37°C for 1 h, and the plates were then washed 5 times. Subsequently, the GtfB amount or antibody titer was assessed using an alkaline phosphatase substrate kit according to the manufacturer's instructions (Bio-Rad).
Protein assay.The amounts of GtfB or protein in antiserum were estimated using a Pierce bicinchoninic acid protein assay kit according to the supplier's manual (Takara Bio Inc., Shiga, Japan).
PCRs.All PCR analyses in this study were performed using ExTaq Hot Start version (Takara) according to the manufacturer's instructions as described previously (18). In brief, PCRs were performed using 30 cycles of a denaturing step at 98°C for 10 s and a primer-annealing and extension step at 60°C for 30 s or 1 min, according to the amplicon size.
Prediction of antigenic regions of GtfB in silico.The antigenic regions of GtfB were predicted using Kolaskar's method (21; available at http://imed.med.ucm.es/Tools/antigenic.pl) or Parker's method (33) and Welling's method (49) using ANTHEPROT 2000 version 6.0 software (7; http://antheprot-pbil.ibcp.fr/index.php).
Preparation of recombinant CAT and GBD and generation of anti-CAT and anti-GBD antisera.Based on the results with Kolaskar's method, PCR primers to amplify DNA fragments encoding CAT and GBD were designed (Table 1). The pSK6 plasmid that harbors the gtfB gene from S. mutans MT8148 (13) was used as a PCR template. Each amplified fragment was digested using the restriction enzymes that were added to the PCR primers and cloned into pGEX-6P-1 (GE Healthcare). The recombinant CAT and GBD were expressed as glutathione S-transferase (GST) fusion proteins. These recombinant proteins were digested by passing through PreScission protease (GE Healthcare), and their GST regions were removed using a GSTrap FF column (GE Healthcare). Recombinant proteins and the adjuvant were used to immunize the rabbits, and anti-CAT and anti-GBD antisera were obtained as described above.
Construction of DNA vaccine plasmid and VR adenovirus.The variable region (VR) containing the antigenic regions predicted by Parker's method was amplified by PCR using the pSK6 plasmid as a template and the primers shown in Table 1. The amplicon digested by KpnI and PstI was cloned into pSecTag2B (Invitrogen), and pSecTag2B-VRGB was constructed as the DNA vaccine plasmid for VR (Fig. 1A). The expression cassette was cleaved by NruI and SphI from pSecTag2B-VRGB, blunted, and used to construct the recombined adenovirus according to methods described in a previous report (44). In brief, the blunted expression cassette was ligated with SwaI-digested pALC3 vector and introduced into E. coli DH10B by using an in vitro λ phage packaging kit (Nippon Gene). The structure of the resulting cosmid, pALC3-VRGB, is illustrated in Fig. 1B. To generate infectious recombinant adenoviral vectors, the pALC3-VRGB cosmid and pOG44 plasmid (Invitrogen) for the expression of FLP recombinase were cotransfected into HEK 293 cells (293 cell) seeded on a gelatin-coated well by using Lipofectamine (GIBCO-BRL) according to the supplier's instructions, and the recombinant adenovirus AdV-VRGB was harvested after 15 days from wells in which bacteriolysis complement was observed (Fig. 1C and D). The titer of the harvested adenoviral suspension was measured using an Adeno-X rapid titer kit (Takara).
Detection of expression of the AdV-VRGB-mediated gene and protein in 293 cells.Total RNA was extracted using TRIzol reagent (Invitrogen) from the 293 cells 3 days after they were infected with AdV-VRGB, and reverse transcription (RT) was performed using SuperScript III (Invitrogen) and oligo(dT)20 primer (Invitrogen). To detect the mRNA of AdV-VRGB-mediated genes, we used primers to amplify the VR (Table 1) and β-actin primers (Invitrogen) as an internal control. In addition, total protein was extracted from the 293 cells 3 days after AdV-VRGB infection and was assessed by Western blotting and enhanced chemiluminescence.
Immunization with DNA vaccine for VR and generation of anti-VR antiserum.The DNA vaccine for VR was used to immunize 6-week-old female mice (BALB/c AnNcrlCrlj; Charles River Laboratory Japan, Inc.) 4 times every night using retrograde intracommon ductal injections (43). The mice were injected with AdV-VRGB suspension (1 × 109 PFU in 200 μl of lactated Ringer's solution) the first time according to methods described in a previous report (43) and with 50 μg of pSecTag2B-VRGB plasmid in lactated Ringer's solution the other 3 times. The antiserum titer was measured by ELISA using 10 ng of the purified GtfB as an antigen, and serum was harvested from ascites fluid 2 weeks after the last immunization.
Inhibition assay of GtfB activity by anti-VR antiserum.The effect of anti-VR antiserum on GtfB was analyzed according to a previous method (11). A solution of purified GtfB (1.5 μg/μl) was incubated with an equal volume of anti-VR antiserum, anti-CA-Gtf antiserum, or sham serum undiluted or diluted 1:3 in PBS. The reaction mixtures of GtfB and serum were then incubated with [14C]glucose-sucrose at 37°C for 1 h, and the amount of [14C]glucan was measured as described above. Triplicate samples were analyzed in this experiment. Significant differences were determined using Student's t test (P < 0.05).
Purification of GtfB.The cell-associated GtfB and GtfC enzymes were extracted from S. mutans MT8148 cells by using 8 M urea and were purified by DEAE-anion-exchange chromatography and the following hydroxyapatite column chromatography (Fig. 2A and B). In DEAE-anion-exchange chromatography, Gtf activity was observed in fractions 6 to 17, which were recovered as active fractions. During hydroxyapatite column chromatography, GtfB and GtfC were eluted in fractions 29 and 24, respectively. SDS-PAGE of GtfB and GtfC gave single protein bands with molecular masses of approximately 165 and 163 kDa, respectively (Fig. 2C). Anti-CA-Gtf antiserum reacted with both GtfB and GtfC (Fig. 2D), and anti-Gtf-I and anti-Gtf-SI antisera reacted with GtfB and GtfC, respectively (Fig. 2E and F). The anti-Gtf-I antiserum cross-reacted slightly with GtfC. The purified GtfB was used in further experiments.
Predicting antigenicity using Kolaskar's method and the practical antigenicity of CAT and GBD derived from GtfB.As shown in Fig. 3, primary catalytic and glucan-binding sites were included in the group of highly antigenic candidates. On the basis of this prediction, we designed PCR primers for CAT such that the primer could amplify the region, including primary and secondary catalytic sites. Likewise, the PCR primers for GBD that could amplify the region, including the 6 repeating units of GBD, were designed. The CAT and GBD region fragments amplified by these PCR primers were ligated into pGEX-6P-1 to form pGEX-6P-1-CAT and pGEX-6P-1-GBD, respectively. The recombinant CAT and GBD were expressed as GST fusion proteins, and the GST regions were removed from them by using PreScission protease (Invitrogen). The recombinant CAT and GBD proteins were then used to immunize rabbits, from which anti-CAT and anti-GBD antisera were obtained. The lysates from E. coli recombined with pGEX-6P-1, GEX-6P-1-CAT, and pGEX-6P-1-GBD were electrophoresed by SDS-PAGE and observed as 26, 38, and 72 kDa of GST fusion proteins, respectively (Fig. 4A). In the Western blot analyses used to evaluate the antigenicity of CAT and GBD, it was observed that anti-CA-Gtf antibody did not react with the CAT or GBD proteins (Fig. 4B), while anti-CAT and anti-GBD antisera reacted with the corresponding recombinant proteins, and the native GtfB served as a positive control (Fig. 4C and D). In the inhibition assays of these antibodies (Fig. 5), anti-CAT and anti-GBD antibodies were less able to inhibit GtfB activity than was anti-CA-Gtf antibody, although there was a significant difference between the glucan synthesis inhibited by the anti-GBD antibody and the sham serum. Thus, it was suggested that the CAT and GBD regions are not immunodominant compared with the other antigenic regions that react with anti-CA-Gtf antibodies, although they are antigenic in rabbits.
Reevaluation of the antigenic region in GtfB by using ANTHEPROT software.As it was suggested that CAT and GBD were nonimmunodominant, we reevaluated the antigenic region of GtfB by the other in silico predictions identified by the Parker and Welling methods by using the ANTHEPROT software (Fig. 6).
According to these analyses, the antigenicity of the primary catalytic site was predicted to be low by both methods, as this region was highly hydrophobic, lowly hydrophilic, and had solvent accessibility. On the other hand, the GBD region was also predicted to have relatively low antigenicity by both methods, although the 6 periodic antigenic peaks corresponding to the 6 repeating structures were identified as antigenic epitopes in Parker's prediction.
When GtfB was roughly divided into 3 parts, VR, CAT, and GBD, Parker's prediction indicated that the N-terminal VR had more antigenic peaks and was more solvent accessible than the other 2 parts. No report has evaluated the antigenicity of VR from GtfB. Thus, this region was adopted for use in the following experiments.
Expression of AdV-VRGB-mediated gene and protein in 293 cells.In the RT-PCR analysis used to confirm AdV-VRGB-mediated mRNA expression, an approximately 1.1-kbp fragment was amplified in the sample from the Adv-VRGB-infected 293 cells (Fig. 7A). Western blot analysis was then used to confirm that the protein derived from AdV-VRGB was produced; an appropriate protein band with a molecular mass of approximately 46 kDa, including a myc epitope and histidine tag derived from pSecTag2B, was observed in the AdV-VRGB-infected 293 cells (Fig. 7C).
Immunization of mice with AdV-VRGB and pSecTag2B-VRGB.To induce the development of antibodies against VR, AdV-VRGB and pSecTag2B-VRGB were injected at the back of the hind legs of mice once and thrice, respectively. The reason why AdV-VRGB was not used for all 4 immunizations was to prevent the mice from dying due to the injection of adenovirus. To measure the titer of anti-VR IgG, ELISA using the purified GtfB as the antigen and goat anti-mouse IgG antibody as the secondary antibody was performed. It was observed that the titer of anti-VR IgG increased in 2 of 5 mice (Fig. 8). Thus, it was revealed that immunization with AdV-VRGB and pSecTag2B-VRGB could induce the development of antibody against VR. The antiserum against VR recovered from these 2 mice was pooled and used in further experiments.
Evaluation of the antiserum induced by DNA vaccine against VR.Reactivity of the antiserum induced by DNA vaccines against VR was evaluated by Western blot analysis using crude CA-Gtf extracted from S. mutans cells, using urea as an antigen (Fig. 9). The results showed that the antiserum induced by DNA vaccines against VR reacted with natural GtfB. Thus, it was suggested that the VR of GtfB is the antigenic region and is more reactive than CAT and GBD.
Inhibition of GtfB activity by anti-VR antiserum.We carried out an inhibition assay to prove that the anti-VR antiserum from mice contains antibodies that react with a native protein such as anti-CA-Gtf and to evaluate the effect of anti-VR antiserum on the glucan synthesis of GtfB (Fig. 10). In this experiment, the protein concentration of each nondiluted anti-VR antiserum, anti-CA-Gtf antiserum, and sham serum was 60 μg/μl. When the ELISA using equal volumes of GtfB (concentration, 1.5 μg/μl) was performed, the absorbances at 405 nm of the nondiluted anti-VR antiserum, anti-CA-Gtf antiserum, and sham serum were 1.2, 1.6, and 0, respectively. These results revealed that the anti-VR antiserum significantly inhibited GtfB activity compared with the sham serum. The anti-VR antiserum inhibited GtfB activity almost to the same degree, as did the anti-CA-GtfB antiserum when 60 μg protein of the antiserum was used. Thus, these results suggested that the anti-VR antiserum induced by AdV-VRGB and pSecTag2B-VRGB effectively inhibited GtfB activity.
In the present study, we reevaluated the effectiveness of the CAT and GBD as epitopes in vaccine production and also found that the ca. 360-amino-acid VR, which exists at the N terminus of GtfB, is a more reactive vaccine target.
The large Gtf molecules hamper complete functional domain mapping. However, studies comparing the amino acid sequences of Gtf from various oral streptococci (Fig. 6) revealed that Gtf consists of 4 regions: a ca. 40-amino-acid conserved signal sequence; a ca. 360-amino-acid VR with unknown function that is species specific and not conserved among the other Gtfs; a ca. 500-amino-acid catalytic domain that contains conserved amino acids that are necessary for sucrose hydrolysis (14, 25, 27); a series of 6 direct repeats that function in glucan binding (1, 9, 28). The latter 2 functional domains were employed as potential vaccine target regions in previous attempts to develop a component vaccine against GtfB, owing to their association with enzyme function and the high degree of sequence conservation among streptococci. However, the antigenicity of those 2 regions was confirmed only by Western blot analyses using antibodies induced by synthetic peptides corresponding to the CAT or GBD regions derived from GtfI of Streptococcus downei (40, 41). Thereafter, to make up for antigenicity, the diepitopic antigen consisting of both the CAT and GBD (45), the chimera of CAT and the B subunit of cholera toxin (22), and the chimera of GBD and the saliva-binding region of Ag I/II (50) were applied. On the basis of these successful reports, we also tried to develop a DNA vaccine against CAT and GBD, but we did not obtain the expected results. One result was that the recombinant CAT and GBD did not react with the antiserum induced by native Gtf from S. mutans (Fig. 4), although these recombinant proteins did induce antibodies that reacted with native GtfB and the corresponding recombinant proteins. In addition, the DNA vaccine plasmid for CAT and GBD using pcDNA3 did not mediate sufficient amounts of the recombinant proteins to induce specific antibodies, although expression of the objective mRNA was confirmed by RT-PCR in an in vitro experiment (data not shown). Thus, we began to doubt the antigenicities of these regions as a way to express antigenic protein and decided to focus on alternative domains.
Secondary structure predictions suggest that the Gtfs are members of the α-amylase superfamily and contain a circularly permuted (α/β)8-barrel motif (8, 19, 26). Our results in Fig. 6 revealed that the primary CAT site (FDSIRVDAVDN) of GtfB is highly hydrophobic, and its solvent accessibility was very low. Since these observations suggested that the primary CAT site existx inside the (α/β)8-barrel structure, it was thought that this region may be difficult to recognize as an antigen for the immune system, although low-molecular-weight sucrose is able to pass through this structure as an enzyme substrate. Thus, it was suggested that the antigenicity of CAT would be low, as shown by Parker's prediction.
In our previous study of glucosyltransferase from Streptococcus oralis (GtfR; 11), the incomplete recombinant GtfR without GBD reacted with the antiserum induced by native GTFR from S. oralis. An antiserum preliminarily prepared by immunization with the recombinant GtfR without GBD not only reacted with native GtfR in Western blot analysis but also inhibited glucan synthesis by GtfR (data not shown). On the other hand, GbpA and GBD are homologous, and it was reported that immunization with the entire GbpA protein did not elicit a protective immune response (2, 36). Taken together, these observations indirectly suggest that GBD antigenicity is lower than that of the other Gtf regions. Practically, our results suggested that GBD would not be immunodominant, because few antigenic peaks were seen compared with the other GtfB regions according to the analysis using Parker's method (Fig. 6).
In this study, we reevaluated the antigenic GtfB region and tried to find a more potent antigenic component. Results of the in silico analysis using Parker's method suggested that the VR derived from GtfB was immunodominant, as evidenced by its many antigenic peaks compared with those of CAT and GBD (Fig. 6). As it was reported that the VR is specific to the Gtf of individual Streptococcus species (17, 18), our DNA vaccines would induce GtfB-specific antibodies that would have an effect only on S. mutans glucan synthesis and colonization and would not interfere with Gtf-expressing members of the oral commensal microbiota.
It was recently reported that nonimmunodominant regions were effective as building blocks in a streptococcal fusion protein vaccine (42). Although CAT and GBD were suggested to be nonimmunodominant in this study, they nevertheless may be attractive vaccine targets. Thus, fusion proteins of VR and CAT or GBD may induce more effective antibodies to inhibit glucan synthesis of GtfB.
We adopted the adenovirus vector, which possesses an intensive promoter and a high infectious ability, as a way to deliver recombinant antigen protein sufficient for mucosal immunization as well. Since it was demonstrated that AdV-VRGB expressed sufficient recombinant antigen protein to induce the development of antibodies that could inhibit GtfB activity in murine in vivo experiments, use of AdV-VRGB may also be effective with mucosal route immunization. As a natural consequence, AdV-VRGB ought to be applied in oral experiments in animals to test whether dental caries can be prevented. We realize that there is a safety problem when it comes to application of this DNA vaccine virus. Even if it were safe, its use would not be permitted in the human oral cavity for dental caries prevention. Because of safety issues, we hesitated to use only DNA vaccine virus in all immunization steps; instead, we used DNA vaccine plasmid in the last 3 immunizations to reduce stress in the mice. We plan to use AdV-VRGB to make a hybridoma that produces anti-VR antibody to clone the gene encoding anti-VR antibody by phage display (35) and to transform this gene into a rice plant by using Agrobacterium-mediated transformation (31, 32). Rice harvested in this planned study will be evaluated for passive immunization to prevent dental caries.
We thank T. Mitsu and K. Kinugasa of the Oriental Yeast Co., Ltd., for their technical support concerning the adenoviral experiments.
This work was supported by KAKENHI (Grant-in-Aid for Scientific Research) from the Japan Society for the Promotion of Science (21659477).
- Received 18 March 2011.
- Returned for modification 12 April 2011.
- Accepted 5 July 2011.
- Accepted manuscript posted online 27 July 2011.
- Copyright © 2011, American Society for Microbiology. All Rights Reserved.


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Mia Winters was a perfectly normal junior at Yale University. Until she accidentally witnessed a secret lovers' rendezvous. Now, blackmailed into silence, a sexual hunger has awoken inside of her, and there's only one way to satiate it–but she's not sure she wants to. Of course, Julian Roth, her gorgeous and brilliant T.A., and Tristan Masters, her equally gorgeous but loathsome classmate, may not give her a choice. The closer she gets to them, the more she is drawn into the world of old money, family dynasties, and secret societies–all worlds she never intended to discover. Will she let herself go down the proverbial rabbit hole or will her better judgment win out in the war between what her brain thinks and what her body craves?
This work of fiction is erotica, meaning that sexual encounters are descriptively explicit and extensive. This novel features a "MMF" relationship, meaning two men and one woman are all involved in a three-way sexual relationship. Content is inappropriate for non-adults.
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Anna James Watson lives in Kennewick, Washington, with her husband, daughter, and more four-legged critters than she cares to admit. She writes for love, drinks too much tea, and is deeply addicted to sleep. Her patronus is a manx cat, and her wand is Cedar, 12 and 3/4 inches, Unicorn Hair core, unbending flexibility.
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Hi there! I'm a mom and wife by day (and night), aspiring writer (in my dreams), and an avid reader (every day of the week). This is my little corner of the web to post about ALL things books. I live in Copenhagen, but I am a Canadian girl (born and raised) at heart.


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In this article
I walked out of my marriage with a mountain of debt on one hand, and a one-year-old boy on the other.
My first decision was to move in with my parents despite having to commute from Klang to Kuala Lumpur for work daily. Living with my parents didn't just mean having a roof over my head and supportive embraces when I needed it. It meant I could save on housing and daycare, helping me direct my savings towards clearing my debt. My father helped by keeping me motivated about my job and I had my mother whom I could trust to take care of my son.
The first two years was a trying time filled with personal sacrifices, but not only did I survive, I think I thrived.
Debts and financial burdenDuring my marriage, I had incurred a lot of debt, the result of maxed out credit cards and a personal loan that I had taken for my ex-husband. Also, I had to bear the cost of my car's numerous repairs, from accidents caused by my ex-husband's reckless driving.
The legal fees from my divorce was RM8,000. My car loan, study loan and baby's medical fees amounted to about RM2,000 a month. Also, my son Harshad was prone to eczema and food allergies which led to expensive hypo-allergenic infant formula, medication and skin care.
Though I was earning well as a corporate training manager, I struggled to make ends meet as my expenses kept piling up alongside my debts.
Financial baby stepsThere was no instant miracle solution or an easy way out of my financial situation so I took things one step at a time. My father helped with organising my finances and together we worked around what we had.
First, I decided to consolidate all my credit card debts. After I explained my situation, the credit card company offered me a personal loan to pay off the credit card charges and offered me fixed instalments with lower finance charges.
I also got help from my friend and financial planner, Helen. I had been contributing to a mutual savings fund for a couple of years. Helen sold a portion of my fund and advised me on how to prioritise my debt clearance.
I also needed to manage my legal fees. Luckily, the firm agreed to my request to pay off my legal fees in four instalments.
As for my car, I was able to sell it. My father bought a new car and gave me his old but reliable Proton Waja, thus reducing my monthly expenses.
Living on a budgetAs my priority was to first settle all my debts, there were a lot of lifestyle changes that my family and I had to make. Shopping was restricted to necessary items for my child and household. We had all our meals at home with only the occasional treats at restaurants on special days such as birthdays. We hardly went for any outings, not even to the movies. I considered going shopping at the pasar malam a leisure activity.
I made cutbacks in as many areas as I could, right down to little things such as going to office early (I left home at 6.30am everyday) and stayed in the office till after peak traffic hours just to save on petrol and to avoid paying toll. I also packed food from home as often as I could.
Close friends and relatives were a great source of strength, reliance and moral support. I did not get to celebrate my son's first birthday due to marital issues and fights during my marriage. However, with help from my family and friends, we made sure his second birthday was a memorable one.
I took in part-time work by doing some training facilitation and writing research papers for trainers. All these brought in extra income that helped me out a lot.
Being an independent person, falling back on my parents for financial assistance was awkward at first, but I have learnt that accepting help sometimes goes a long way. I am an only child and understandably my parents were deeply saddened by my circumstances. However, they remained strong and supportive.
I'll always be thankful to my family.
Moving forward and thrivingGradually, I managed to pay off my debt and things started looking brighter. I even managed to establish a more civil relationship with my ex-husband, and he has agreed to contribute monthly to our child's expenses.
I'm currently pursuing my Masters. This move boosted my confidence as it is a sure means toward career advancement. I believe education will spur career growth and land me in a better financial position. I feel this is essential to look after Harshad as well as my ageing parents' well-being.
Now with greater financial stability, my parents, Harshad and I are in the midst of relocating from Klang to Petaling Jaya. Our new home is closer to my work place, so I can save on precious time commuting and in the long run, save on fuel cost and toll charges. It would also mean more quality time with Harshad.
We have come a long way, my child and I, with the support of my parents. I still budget these days as it helps me keep tabs on my spending. Now, I am also able to set aside at least twenty per cent of my salary towards an insurance policy and savings account for Harshad's future.
I hope someday I can repay my debt to my parents, without whom, I will not be the person I am today.
Have you got a story to tell about your parenting journey? Share it with other mums and dads in our Community.


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Search the Community
Showing results for tags 'surfacebook 2'.
Found 1 result
Hey, I recently started the trial to see if I would like this program and so far I have, however, one thing that I noticed that will really bother me is that with the surface pen (the one with the top and side button) I can't press the right button and use the pencil's color picker. Is there a way to remap that or is there a tablet mode I can use to make it work better in the tablet mode on my surface book (Like photoshop's feature to use all the hotkeys on the left)? Thank you guys in advance, if one of those features are in there, I will most definitely get the program


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Use this guide for all of your Philosophy information needs. Search the Library Resources below for books, eBooks, articles, and streaming videos. Use the tabs above to explore web sites and use citation tools.
image from Wikimedia Commons: Political Philosophers Left to Right: Plato, Aristotle, Thomas Aquinas, René Descartes, David Hume, Immanuel Kant, G.W.F. Hegel, Friedrich Nietzsche, Gottlob Frege and Edmund Husserl
Search for books and movies in CCAC's libraries. We also have an advanced search screen. Show me how
Use the "Place Hold" feature to have items owned by another campus sent to your campus. This process can take 3 or 4 business days. Show me how


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ZonesCorp, one of the largest developers of economic zones in the UAE, has announced a new agreement with Saudi Arabia's Berain Water to build a AED138 million ($37.5 million) water bottling plant in the food industry zone of Al Ain Industrial City.
The announcement comes as the food manufacturing sector in Abu Dhabi has witnessed "remarkable growth" over the past few years as a result of the booming domestic market and growing exports, ZonesCorp said in a statement.
Saeed Eisa Mohammed Al Khyeli, director general of ZonesCorp, said: "The UAE and Saudi Arabia enjoy strong historic relations, and our latest partnership with the Saudi company Berain Water testifies to the strength of those ties."
Mahdi Nasser Al-Qahtani, CEO of Berain Water, said that the facility is set to commence operations in the first quarter of 2020, adding that it will produce around 70,000 bottles per hour.
The facility will also create 424 jobs in manufacturing, marketing, distribution and sales which may increase to 807 jobs within two years with the addition of new production lines in 2021, he said.
ZonesCorp has developed dedicated zones for the food industry that span areas in Abu Dhabi and Al Ain. The two cities currently host several operational facilities, ranging from medium to large factories that produce bottled drinking water, soft drinks, dates, animal feed, poultry products, dairy products and other food items.


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Julie, you know it all! Julie, how did you know that? Julie, you're amazing. Julie, I want to be you.
OK, so no one really goes that far! My goal today is to show you where I get all my information so that you can use it too. By no means is my list a complete compilation of all wellness things on the internet or even a total list of everything I read, but you might find some interesting blogs that you'd like to follow. Am I telling you to read them instead of me? Well, hopefully you have time for both 🙂 Since I'm just starting this blog and I'm potentially advising you on things – fitness, nutrition, self care, wellness -I thought you needed to know where I'm getting some of my information.
When you read my blog remember that my intention isn't to put out misinformation. And, this isn't a forum for me to be right 100% of the time for all of eternity (although, if you're my husband, you think I am). New information comes out and I want to keep up with that so you get the best information possible. My intent is to write about what is working for me and my clients. There will be occasions when I write a post and later I change my mind or find new evidence that something else works better or I find out that I was completely wrong. I'll make sure to put you first and let you know what's happening. Me being right isn't above your wellness.
Now is a good time to say that it's important to recognize that I don't agree or follow all of the advice from the sites listed below. I take what works for me and move forward incorporating it into my life. You will have to do the same – not everything works for everyone. Find your answers, find your truth, define your health. My goal is to give you some resources along the way.
So, here's a list of resources. These aren't all blogs, but if I'm reading a book by one of the enthusiasts below, I've included a link to their page. There's also a podcast and some online tv. I'm open to anyone adding to this list – I know there is so much more out there and like you, I'm still figuring out my health.
In no particular order…
- www.idoportal.com/
- www.rawbrahs.com/
- www.mobilitywod.com/
- http://breakingmuscle.com/
- http://www.barbellshrugged.com/
- http://www.anatomytrains.com/
- www.getbodysmart.com/
- http://bretcontreras.com/
- http://www.jensinkler.com/
- https://www.dellanave.com/
- balancedbites.com/
- chriskresser.com/
- paleoparents.com/
- www.marksdailyapple.com/
- startingstrength.com/
- www.t-nation.com/
- www.sethgodin.com/
- www.primalpalate.com/
- mollygalbraith.com/
- www.ericcressey.com/
- www.louschuler.com/
- http://iamerinbrown.com/
- strengthofevidence.com/
- www.tonygentilcore.com/
- www.functionalmovement.com/
- http://www.how-to-meditate.org/
- http://www.gaiamtv.com/


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The Cutest Little Angel Costume you will ever see. Stretchy white lycra bodice with a peek-a-boo panel at the front and double halter neck straps.
The skirt is covered with three rows of zig-zag white netting and is edged with elaborate lace.
Also included are wired netting wings, covered with gold glitter stars (they hold on with elastic straps), and a headband with fluffy feather covered halo.


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license soup art for commercial use
Search and Download High-Quality Soup Content Straight from Social Media
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Bank is financial institution which provides financial services to the customers. Basically the institution that's provides fundamental services like accepting deposits and providing loans.
MICR is defined as the Magnetic Ink Character Recognition Code. This code is used by all the financial institution in India for processing cheques. Know about the Almora Urban Co-Operative bank's MICR code in this post.
You may not know, there are different kinds of banks are in India and not every banks are authorized to do same thing. In this article, I am going to discuss about commercial bank and its role and fun
Bank is the financial institution providing financial support for their account holder. According to their nature, banks are classified into three types. Public Sector Banks Private Sector Banks Foreign banks. Let us discuss about each of them in detail
Now a day's online banking plays the major role in fund transferring and capital market transactions. Net banking users are rapidly increasing, to make the transaction easy and less time. Let us discuss how to secure it in this post
There are totally 165 banks and more than 1,20,000 branches are providing financial support for their account holders in India. All the branches has unique 11 character IFSC and here is a post to help you find the IFSC Code for PNB and SBI banks in India
RBI (Reserve Bank of India) is the central bank which has the power to control all the banks in India. Reserve bank plays a vital role in our Indian economy. In this article we are going to see about how printed currency enter into market and how it works.
Visa and Master Card are the two leading credit card companies in the world. These both companies are operating in similar way. Visa has issued 1.6 million cards and the MasterCard has nearly 18 million users. In this post, let us discuss the different credit card providers.
Mobile Banking is the one of the most common facilities provided by all the Banks in India. It not like the Internet banking but you can check your balance. Let us now discuss in detail about the mobile banking.
Banking Fraud are the posing thread of Indian economy. In India there are 347 cyber-crime in the year of 2004, where the number is rapidly increased to 481 in 2005. Don't worry, this post will help you to know how to prevent your account from banking fraud
Do you enter wrong IFSC Code while money transaction either offline or online? Don't know what will happen next? Here is a post to help you know what will happen if you enter wrong IFSC code during a transaction.
All the financial institution in India are using 9 digit MICR code for processing cheque. Bank users should know the purpose of the MICR code. This post will help you to know the details about the IFSC and MICR Codes.
Do you know about the most important bank's code, the swift code? Don't worry, in this post I am going to explain to you about the Swift code and why it is required.
Indian Bank is one of the famous public sector banks which was established in the year of 1907 in Madaras (Chennai). There are 2537 branches in 33 states of India. There are many inquiry options and here is a post to help you with it.
Do you know you can check your bank's account balance through just a single missed call? Don't worry, this post will guide you through the steps to do it.
The banking concept in India is introduced in half of the 18th century and the first bank was The General Bank of India established in the year of 1786. Following that State Bank of kolkata was founded in the year of 1806 which was renamed as The Bank of Bengal.
This post will explain you the different ways of paying Axis bank credit card bills online. Pay your bill through online to save your time and energy.
There are number of ways to pay your Axis bank credit card bills either online or offline. But in this post, let us discuss about the credit card bill payment through offline.
India: One of the fast developing democracy countries in the world. Moreover, India is the second largest populated country in the world. Here is a huge list of debit cards and bank accounts for kids in India.
Bank information Blog. Here you can get recent banking information, internet marketing tips etc
In India, there are plenty of banks are available which is combination of Public, Private and Cooperative sectors banks. When you are initiate money transfer to anyone in the world is most important. Know about the Demonetisation with this post.
Here, you can see what is the IFSC Code and ways to find IFSC code.
Cash Deposit Machine (CDM) will look like an Automatic Teller Machine (ATM), but both have different functionalities. In CDM you can deposit money without interaction of humans like a counter. Know more about CDM with this post.
Have you forgotten your internet banking password? Don't panic. You can easily reset your lost password. This post will guide you through some simple steps to help you reset your internet banking password for axis bank.
After, Opened a new saving account, you will get a Chequebook in the welcome kit. It has ten Leaf. If it is not sufficient, then this post will help you to apply for a new cheque book.


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Max Mineral Naked Broad Spectrum SPF 45
Peter Thomas Roth
Peter Thomas Roth
Estimated Delivery : Oct 29 - Oct 30
FREE 2-day shipping on all orders within the US.
Returns are FREE in the US via:
- In person with Happy Returns
- The mail
Returns are accepted within 30 days of purchase for a full refund and 60 days for an exchange or store credit. Restrictions apply.
Return within 30 days if unopened and unused
Sun protection you can't even feel: Peter Thomas Roth Max Mineral Naked Broad Spectrum SPF 45 Lotion. This lightweight, all-mineral lotion is formulated with Zinc Oxide and Titanium Dioxide for broad spectrum SPF 45 protection, while Vitamins A, C, and E provide antioxidant benefits. The vanishing universal tint includes light-scattering optics from diamond powder, leaving a matte finish with a natural glow.
The Details
- Safflower Extract nourishes and provides antioxidant benefits
- Silica, a natural mineral, helps to absorb excess oil
- Layers beautifully over moisturizer and under makeup
- 1.7 fl oz
- Revolve Style No. PTHO-WU36
- Manufacturer Style No. 31 01 009
- Actual packaging and materials may vary. Please read labels, warnings, and directions before use.
How to Use
- Apply to face and neck liberally 15 minutes before sun exposure
- Reapply at least every two hours
Water/Aqua/Eau, Butyloctyl Salicylate, C9-12 Alkane, Carthamus Tinctorius (Safflower) Seed Oil, Dicaprylyl Carbonate, Isodecyl Neopentanoate, Cetyl Alcohol, Coco-Caprylate/Caprate, Propanediol, Ethylhexyl Olivate, Glyceryl Stearate, Peg-100 Stearate, Cetearyl Olivate, Cetearyl Phosphate, Triethoxycaprylylsilane, Phenoxyethanol, Sorbitan Olivate, Oleth-3 Phosphate, Hdi/Trimethylol Hexyllactone Crosspolymer, Peg-8 Dimethicone, Alumina, Xanthan Gum, Polymethylsilsesquioxane, Polyhydroxystearic Acid, Disodium Edta, Squalane, Gluconolactone, Isopropyl Titanium Triisostearate, Sodium Benzoate, Hydrogen Dimethicone, Potassium Sorbate, Silica, Mica, Ascorbic Acid, Octyldodecanol, Retinyl Palmitate, Sodium Propoxyhydroxypropyl Thiosulfate Silica, Tocopheryl Acetate, Diamond Powder, Iron Oxides (Ci 77491, Ci 77492, Ci 77499).
If an item you would like to purchase is out of stock, you can place a Special Order Inquiry. REVOLVE will then check the availability and obtain an approximate arrival date for the item, if available. Typically, it takes up to 3-4 weeks to verify this information.
Placing a Special Order Inquiry is absolutely free and there is no obligation to follow through with a purchase.
- You will be notified by email once we receive the item.
- You may cancel at any time without penalty
- Your card will ONLY be charged when your order is shipped.
- Availability is not guaranteed.


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DIY cleaning products
Collection by Sambofett
How To Refresh and Clean Your Pillows | The Homestead Survival
Learn how to refresh and clean your pillows. Cleaning and refreshing your pillows three times a year is a way to remove dust, mold, bacteria, dead skin
How to Make Disinfectant Wipes / Reusable DIY "Clorox" Wipes
If you want to learn how to make disinfectant wipes, then you will love this recipe for REUSABLE DIY Clorox Wipes! They even smell lemony-fresh!
Mom's Super Laundry Sauce | Whipped Laundry Soap | Make your own Laundry Detergent
Here's a super simple Whipped Cream Super Laundry Soap, Do It Yourself laundry soap detergent recipe that doesn't leave any residue on clothing, gets nearly every stain known to man out the first time around and smells clean and fresh- oh and did I mention that it costs about $1.76 for 128 loads of laundry?
Downy Unstopable In-Wash Scent Booster Beads, Lush, 20.1 Ounce (Pack of 1)
Buy Downy Unstopable In-Wash Scent Booster Beads, Lush, 20.1 Ounce (Pack of 1) on Amazon.com ✓ FREE SHIPPING on qualified orders
How To Clean Exterior Windows And Have The Cleanest
This article will contain affiliate links, please read my full disclosure policy for more information. If your sitting in the kitchen eating or cooking or maybe typing like me on your computer and look through the window and see dirt on the windows can just ruin the view. Cleaning it can be quite easy if
How to Clean Toe and Foot Marks Off Sandals
Nothing decreases the looks and value of slip-on sandals like toe, heel and foot marks on the insoles. Here's how to remove them.
DIY Jelly Soap is Fun and Easy to Make! (Video)
DIY jelly soap is a great project for kids! Use any kind of liquid soap for endless scent options! Jelly soap makes handwashing fun.
Zero Waste Unpaper Towels
Step by step instructions with photos to make zero waste unpaper towels. It's easy to reduce your paper consumption and save money.
DIY clean your car seats - FunSubstance
Click to see the full post now
The Best Way to Clean Windows
Learn the best way to clean windows with products you probably already have on hand around your home. A super simple method for inside and out! #clean #cleaning #springcleaning #windows
9 Magical Ways to Use Lavender
Lavender has it's own healing qualities for sure, but one it is best know for is it's ability help calm. If you love lavender and are looking for ways to add more of it into your life,…
Reusable Swiffer Duster Cloths Tutorial - Sew Much Ado
Make reusable Swiffer duster cloths with this easy tutorial! No more need to buy expensive refills - you'll love this DIY cloth duster!
Removing Stains from Vintage Linens
Do you have any old family linens packed away that you never use because they have stains or yellowing that you can't get out? Or have you ever come across an inexpensive tablecloth at a yard…
How to Clean Painted Walls From 10 Stubborn Stains | ONE DOES SIMPLY
How to remove stains from the wall? Take a look at these simple cleaning hacks to remove 10 different stains from the walls. See how to clean the walls from pen ink, food stains, black molds, crayon stain, and so many other stains. GO CHECK THEM OUT>>


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Re: Next Up: Maidenhead United (H)
Posted by: Higgosboots (IP Logged)
Date: 24 January, 2018 08:54
They picked up 3 points, that's all that matters for the rest of the season.
Re: Next Up: Maidenhead United (H)
Posted by: TRSince1968 (IP Logged)
Date: 24 January, 2018 11:25
Quote:Hoots MonQuote:853OKGQuote:Hoots MonQuote:853OKGQuote:gateshead 1 Tranmere 0, Macclesfield 2 Tranmere 2, Fylde 5 Tranmere 2. Look who played in those.aktrfcQuote:TranmereFanQuote:HardlyYardley
Good three points. Looking tighter at the top. Will losing 6-0 have a lasting effect on Macclesfield? If we can beat them and Wrexham, we're in with a chance. Credit to Norwood - and let's hope Ginnelly can give him some tips on finishing one-on-ones. Credit to McNulty too for playing with what must be a very sore leg. Think our season may rest on what happens with Wallace and Tollit.
Don't agree with you re:Wallace and Tollit as we are we're we are without them. However will strengthen us for sure.
We are were we are yes. 5th. Agree with hardly, get them back and we stand a much better chance of winning the league. It was Banks and DMH that had the biggest impact on the team. Just look at the results while they were here. Look at the results since, back to scrapping a win at home to a poor team.
Sorry Tranmerefan, but yes we're are were we are without them, but we are were we are with Banks and DMH. We now don't have any of them, so we may be back to were we were before. We need Ben and another asap.
DMH didn't play against Fylde.
Banks did................
Banks arrived less than 48 hours before Gateshead,who were the worst team I've seen us play this season. We were far from great but the very late unlucky own goal was an utter travesty.
And we have won without either of the two. My point is that there are nine other players that make up the team. Clarke needs credit as well. So does Norwood and Norbury and all the others who have scored goals. So I feel the team will adapt because othere players are now playing as part of a team.
I've got a cousin called Norbury but she[or her husband] doesn't play in midfield.
What I typed and what the f in autocorrect replaced it with are two different people.
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- Students
- News & Topics
Note-takers for hearing-impaired students being sought
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Nintendo, as is their policy, stumbled ass backwards into a ridiculously popular product. Their $60 NES Classic Edition was one of the hardest toys to find this holiday season, and the company, thinking that it would only appeal to only the most hardcore retro gamers and not, oh, I don't know, everyone who grew up with a classic Nintendo Entertainment System, didn't make enough to meet demand. So it became incredibly difficult to find them, and they're still not easy to track down.
So it's pretty surprising that today Nintendo announced that they're discontinuing production in North America on the NES Classic Edition. In a statement to IGN, the company said:
Throughout April, NOA territories will receive the last shipments of Nintendo Entertainment System: NES Classic Edition systems for this year. We encourage anyone interested in obtaining this system to check with retail outlets regarding availability. We understand that it has been difficult for many consumers to find a system, and for that we apologize. We have paid close attention to consumer feedback, and we greatly appreciate the incredible level of consumer interest and support for this product.
Now I'm not some fancy-schmancy businessman, but I was under the impression that when people want your product, you keep making it until demand no longer outpaces supply. For Nintendo to pull the plug now makes me wonder what the future plans could entail.
For its part, the company says, "NES Classic Edition wasn't intended to be an ongoing, long-term product. However, due to high demand, we did add extra shipments to our original plans," but to me that seems to say that they stumbled into a popular revenue stream, and one that doesn't conflict with their other products. It makes sense to stop selling Wii U when you've moved onto the Switch, but the NES Classic is going to a completely different group of consumers.
I'm curious to see how the company proceeds in the future. Do they intend to ramp up their virtual console for the Switch? Are they working on an NES Classic 2.0 with more games? Are they (fingers crossed) working on an SNES Classic Edition? It will be interesting to see how Nintendo as well as other companies proceed now that they know there's a lot of demand for high-quality, plug-and-play retro gaming.


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Software Engineer
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The OneDrive/SharePoint group needs you as we tackle deep technical challenges to transition our core productivity apps to the mobile first, cloud first world! We have several positions available at our company headquarters in Redmond and we need Software Engineers with a wide range of skills & experiences.
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We are looking for Engineers who have the desire and drive to work in a fast-paced environment, the architectural skills to build durable platforms, the curiosity to seek out innovative solutions, the agility to work on a broad range of features, and a passion for delighting the customer.
- You will be working on components within the SharePoint Online infrastructure, such as SharePoint data store, capacity management, performance, authentication, networking and much more
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128 - First Day of School—1960, New Orleans
128 - First Day of School—1960, New Orleans
November 14, 1960 — Four six-year-old girls, flanked by Federal Marshals, walked through
screaming crowds and policemen on horseback as they approached their new
schools for the first time. Leona Tate thought it must be Mardi Gras. Gail
thought they were going to kill her.
Four years after the Supreme Court ruled to desegregate schools in Brown v Board of
Education, schools in the south were dragging their feet. Finally, in 1960, the
NAACP and a daring judge selected two schools in New Orleans to push forward
with integration — McDonogh No.19 Elementary and William Frantz.
An application was put in the paper. From 135 families, four girls were
selected. They were given psychological tests. Their families were
prepared. Members of the Louisiana Legislature took out paid advertisements in
the local paper encouraging parents to boycott the schools. There were threats
of violence.
When the girls going to McDonogh No.19 arrived in their classroom, the white
children began to disappear. One by one their parents took them out of school.
For a year and a half the girls were the only children in the school. Guarded
night and day, they were not allowed to play outdoors. The windows were covered
with brown paper.
The story of integrating the New Orleans Public schools in 1960 told by Leona Tate,
Tessie Prevost Williams, and Gail Etienne Stripling, who integrated McDonogh
No.19 Elementary School, and retired Deputy U.S. Marshals Herschel Garner, Al
Butler, and Charlie Burks who assisted with the integration efforts at the schools.
We produced this story a few years back. We want to put it out there again as part of our Keepers Series because it seems critical, particularly now, to remember and pay tribute to the many Keepers of the archives, the stories, the truth about our past and the long fight for what is fair and just.


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Jünger Audio has launched Easy Loudness, a slimline addition to its family of D*AP digital audio processors for loudness control in stereo television and radio broadcast.
With an extensively redesigned web user interface for ease of use and intelligibility, Easy Loudness is available in two versions – one with an integrated 3G SDI interface and the other with a Dante/AES67 AoIP interface. Both versions feature two independent stereo programs which incorporate the Level Magic algorithm to adjust audio from any source, at any time, to the required level without processing artifacts.
Easy Loudness is optimised to run immediately with an established audio connection and requires only minimal configuration. The web interface makes the most important parameters of the operating system directly accessible from the home page, making it very simple to use. In addition to the main interface the device is equipped with an AES/EBU interface to receive a fallback signal. Automatic fail over switching, dual power supplies, SNMP system monitoring, as well as full loudness monitoring and a simple yet powerful snapshot functionality are only fractions of its comprehensive feature set.
In keeping with all the products in D*AP family, Easy Loudness furthers the concept of Smart Audio by allowing broadcasters to invest in simple, reliable and predictable equipment that can automatically deliver controlled audio content while maintaining its quality. Jünger Audio's Smart Audio devices employ the industry standard Ember+ remote protocol, allowing them to seamlessly integrate with an increasingly wide range of compatible equipment including playout automation systems, logging and monitoring processes.
More: www.jungeraudio.com


text: This collection was inspired by nightmares. In our nightmare is where we are mostly inspired. Every look is a different god in their very own universe. The collection should be remembered for being able to step in a room and being powerful without saying much. The colors for the collection was used to show and showcased different concepts power.


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I wish people would have more babies. Not me – I have enough children!! But there's something soul-soothing about maternity sessions for me and I just cannot help myself.
Here on the West Coast, people are having a ton of babies. It's a bit complicated though – we don't have the proper facilities for births, expecting moms have to leave the community to give birth. Recently I attended a luncheon through the Tofino Chamber of Commerce where Clayoquat Biosphere Trust was presenting their most recent Vital Signs report, and I found that particular piece of information so interesting – in 2016 and 2017, 91 and 80 babies respectively were born to West Coast women. Women went to a variety of different places to give birth with Nanaimo topping the list at 60 births. One baby was born in that time frame at Tofino Hospital (and that baby was one of my first clients when I moved to Ucluelet later on in 2017).
Having to leave your community to give birth is a burden for many families. This is a young community full of determined people, and most take it in stride. Many leave and stay with family in the places they themselves are from. Many more take up residence in Port Alberni or Nanaimo for a couple of weeks before their due date.
Sometimes, like in this case, their time on the coast is over and we have to plan their maternity photos for the day before they move away.
We've been spoiled with sunshine lately. The days are hot, which is actually not so good for me because I don't really own a lot of hot-weather clothing and I got a bit of a sunburn the other day while shooting a wedding (WORTH IT though). And yes, I love the golden sunsets – who wouldn't? But there's part of me that's definitely waiting for the fog.
When I was pregnant with my first child – my daughter, who will be 10 this June – we didn't do maternity photos. Photography for me, at that time, was little more than a memory of a hobby (I got my first dSLR when she was 6 months old – prior to that I'd only shot film, and it had been a few years) and maternity photos were only just becoming "a thing". There's basically one photo of me pregnant with her (besides the cellphone selfies – I'll confess to having done those) sitting in our back yard on my yoga mat. Still one of my favourite photos.
With my second pregnancy, I was overwhelmed with photo shoots. I was teaching photography at the time, and wound up as a model for several students, as well as having booked our own family/maternity photos (on vacation – we always had family photos done in Tofino prior to moving out west). Looking back, I love them all, but it was kind of exhausting.
When my friend Sarah Wallace was pregnant (with her second? Third? Fourth? I can't remember) we stomped down to a little creek and she waded in for me – it was amazing. I would love to take an expecting lady up to Virgin Falls one day (soon).
Destiny was a perfect model – I kind of wish she was pregnant all the time (I know she doesn't feel the same). I cannot wait to meet this baby boy when they come to visit.
Guess what? I can't keep this to myself. You know how I've been talking about getting underwater housing? It happened?
And we did an unboxing video.


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Over the past few years, the organisation has adapted to reductions in public spending and changes in the demand for our services. We have been able to do this by changing the way we work but, more importantly, by having a very adaptable and highly-skilled workforce. The services we provide are totally reliant on having well-trained, competent and committed staff.
The Workforce Strategy describes how we plan to train, develop and maintain the competence of our staff across the organisation. It also considers how to deliver enough flexibility in the workforce, in contractual terms, so that we can continue to face the challenges ahead. During 2017, the Authority will be adopting a version of the national workforce strategy for fire and rescue services, which was developed by our Chief Executive.
Our workforce has changed significantly over the past few years. This will continue to happen as we change the way we do things and implement our plans to redesign our services. Our funding has been decreasing over the years and when firefighters or support staff have retired, we have considered carefully whether their role was still required or whether it could be carried out differently.
By streamlining processes and investing in new equipment and electronic systems we have been able to improve efficiency, generate savings and release staff capacity. These improvements have allowed us to maintain, or even improve, the quality of the services we provide despite the reduction in staff numbers. Whilst we will continue to identify efficiencies, we have reached a point where many of our firefighters have reached retirement age and we cannot reduce the numbers any more. This year we will start to recruit whole-time firefighters again.
As demand for our core services declines, there is more reliance on training to ensure firefighters gain the required skills and can demonstrate their competence to do their jobs. As we expand the range of services we provide, for example by attending medical emergencies, we will ensure our staff have the skills they need to make the best possible contribution.
Last year we reviewed the way in which we deliver training to our staff and the outcomes of that review have resulted in the simplification of processes and the removal of the competency recording system. We have started to redesign the way we deliver training and this work will continue in the coming year. Our focus is now on the quality of the training that we offer.
To complement this work we are also developing a leadership programme, since it is important that we enhance and develop managerial skills. These skills will become even more important as we meet the difficult challenges ahead.
We need our workforce to work flexibly if we want to deliver high quality services in an environment where funding is decreasing. In 2014 we started to introduce a flexible rostering pattern of work in some of our stations. The new system was well-received and, since then, we have introduced it to all our day-crewed stations, the fire station at the Channel Tunnel, and our 999 team. Last year we started to introduce this system in our whole-time shift fire stations and will continue with the roll-out this year.
In order to increase flexibility of crewing we have started to use some of our fire engines when only three firefighters are available even though we always aim to have a minimum of four firefighters. We are also planning to review the arrangements for our on-call staff, to make the system more attractive so that more people join the system and stay with us for longer.
As the organisation prepares for the challenges ahead, we know that being successful depends greatly on having a culture fit to face these challenges. In 2015 we conducted an audit of our internal culture and the outcomes have been very useful in helping us define what the culture needs to look like in the future.
We have already started to implement a number of actions to support the cultural change that we need. For example, we have produced a new behavioural framework, have streamlined the internal promotion process, and have simplified the way we record competency. We are also devolving responsibilities more to empower managers. To complement this work a new management and leadership programme is being developed to support managers at all levels in the organisation. We will continue to implement these actions in the coming year.
We care greatly for the health and general wellbeing of our staff. We recognise that sometimes changes at work or issues at home can affect our health and wellbeing. We have introduced a number of tools to help our staff manage their mental and physical wellbeing. We have also produced a training package to help raise awareness of mental health issues. We want to make sure our managers are able to identify and support staff suffering from mental health problems by directing them to the right resources.
Last year we signed up to the Workplace Wellbeing Charter as part of our ongoing commitment to support the health and wellbeing of our staff. The Charter establishes a set of standards relating to the health, safety and wellbeing of employees, against which organisations can assess themselves. In November 2016 we achieved the Excellence Award from the Charter.


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By Peter Reap, J.D., LL.M.
The canned tuna purveyor failed to show that its financial circumstances justified a lower fine—"sorry, Charlie."
The federal district court in San Francisco has ordered StarKist Co. to pay a criminal fine of $100 million, the statutory maximum, for its role in a conspiracy to fix prices for canned tuna sold in the United States, the Department of Justice has announced. StarKist was also sentenced to a 13-month term of probation. StarKist pleaded guilty in October of 2018 for its part in the conspiracy (U.S. v. StarKist Co., Case 3:18-cr-00513-EMC).
According to the one-count felony information that had been filed with the court, StarKist participated in a conspiracy to fix the prices of canned tuna fish from as early as November 2011 through at least as late as December 2013.The conspiracy was an unreasonable restraint of interstate commerce in violation of Section 1 of the Sherman Antitrust Act.
As part of the sentencing hearing, U.S. District Judge Edward M. Chen found that StarKist had not proven that its financial circumstances justified a lower criminal fine. The Justice Department's Antitrust Division opposed StarKist's request for a fine reduction, arguing that StarKist had sufficient financial resources to pay a $100 million criminal fine. In addition to the criminal fine and term of probation, StarKist has also agreed to cooperate in the Antitrust Division's ongoing investigation.
"Today's result demonstrates our commitment to enforcing the antitrust laws aggressively against companies that fix prices," said Assistant Attorney General Makan Delrahim of the Justice Department's Antitrust Division. "Hard-working Americans deserve the benefits of open competition when they spend their hard-earned money on items that stock kitchen shelves. When a corporation cheats customers at the checkout line, the Antitrust Division will hold it accountable to the greatest extent."
"StarKist is committed to being a socially responsible company and doing the right thing. We have cooperated with the DOJ during the course of its investigation and accept responsibility," said Andrew Choe, President & CEO of StarKist in a statement. "We will continue to conduct our business with the utmost transparency and integrity. We have addressed the necessary actions required in this agreement and we will continue to strengthen related compliance best practices."
StarKist was the second company to admit to its involvement in the conspiracy. In May 2017, Bumble Bee Foods LLC agreed to plead guilty to a similar charge.
Companies: StarKist Co.; Bumble Bee Foods LLC
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