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The stock of CubeSmart (NYSE:CUBE) is a huge mover today! CubeSmart (NYSE:CUBE) has declined 17.04% since March 3, 2016 and is downtrending. It has underperformed by 25.38% the S&P500.
The move comes after 6 months positive chart setup for the $4.66B company. It was reported on Oct, 6 by Barchart.com. We have $37.54 PT which if reached, will make NYSE:CUBE worth $2.05 billion more.
Analysts await CubeSmart (NYSE:CUBE) to report earnings on November, 3. They expect $0.37 EPS, up 8.82% or $0.03 from last year's $0.34 per share. CUBE's profit will be $66.14M for 17.61 P/E if the $0.37 EPS becomes a reality. After $0.36 actual EPS reported by CubeSmart for the previous quarter, Wall Street now forecasts 2.78% EPS growth.
CubeSmart (NYSE:CUBE) Ratings Coverage
Out of 11 analysts covering CubeSmart (NYSE:CUBE), 8 rate it a "Buy", 0 "Sell", while 3 "Hold". This means 73% are positive. CubeSmart has been the topic of 17 analyst reports since July 21, 2015 according to StockzIntelligence Inc. The stock of CubeSmart (NYSE:CUBE) earned "Outperform" rating by Raymond James on Friday, November 13. The stock has "Buy" rating given by Evercore on Tuesday, September 8. The stock of CubeSmart (NYSE:CUBE) has "Buy" rating given on Wednesday, November 11 by BB&T Capital. The company was maintained on Thursday, May 12 by Citigroup. The rating was initiated by DA Davidson on Friday, June 24 with "Neutral". The firm earned "Outperform" rating on Thursday, June 23 by FBR Capital. The rating was initiated by Robert W. Baird with "Outperform" on Thursday, July 23. As per Monday, March 21, the company rating was downgraded by Evercore. Jefferies maintained CubeSmart (NYSE:CUBE) on Wednesday, August 10 with "Buy" rating.
According to Zacks Investment Research, "CubeSmart is a self-administered and self-managed real estate company focused on the ownership, operation, acquisition and development of self storage facilities in the United States. Its self storage facilities are designed to offer storage space for residential and commercial customers. CubeSmart, formally known as U-Store-It Trust, is based in Wayne, Pennsylvania."
Insitutional Activity: The institutional sentiment decreased to 0.98 in 2016 Q2. Its down 0.46, from 1.44 in 2016Q1. The ratio fall, as 22 funds sold all CubeSmart shares owned while 96 reduced positions. 39 funds bought stakes while 77 increased positions. They now own 178.71 million shares or 0.94% less from 180.40 million shares in 2016Q1.
The United Kingdom-based Blackrock Group has invested 0.02% in CubeSmart (NYSE:CUBE). Eagle Asset holds 1.63M shares or 0.35% of its portfolio. Citadel Limited Liability Com reported 268,312 shares or 0.01% of all its holdings. Moreover, Schroder Grp has 0.05% invested in CubeSmart (NYSE:CUBE) for 773,178 shares. Spot Trading Ltd Liability Corporation holds 0% of its portfolio in CubeSmart (NYSE:CUBE) for 1,411 shares. The New Jersey-based Quadrant Capital Mngmt Ltd Liability Company has invested 1% in CubeSmart (NYSE:CUBE). Rafferty Asset Management Ltd Company last reported 19,108 shares in the company. Moreover, Barclays Public Ltd has 0.01% invested in CubeSmart (NYSE:CUBE) for 198,377 shares. Nationwide Fund Advsrs accumulated 0.02% or 115,680 shares. Tdam Usa holds 41,185 shares or 0.06% of its portfolio. Axa reported 4,046 shares or 0% of all its holdings. Moreover, Brown Brothers Harriman & has 0% invested in CubeSmart (NYSE:CUBE) for 183 shares. Marshall Wace Ltd Liability Partnership holds 0.01% of its portfolio in CubeSmart (NYSE:CUBE) for 25,400 shares. Dimensional Fund Advisors Lp last reported 0.04% of its portfolio in the stock. The Texas-based Next Grp has invested 0.01% in CubeSmart (NYSE:CUBE).
Insider Transactions: Since August 31, 2016, the stock had 0 insider buys, and 2 insider sales for $1.18 million net activity. DIEFENDERFER WILLIAM M III also sold $80,689 worth of CubeSmart (NYSE:CUBE) on Friday, September 9. On Wednesday, August 31 KELER MARIANNE M sold $1.10 million worth of the stock or 40,000 shares.
Another recent and important CubeSmart (NYSE:CUBE) news was published by Forbes.com which published an article titled: "Ex-Dividend Reminder: CubeSmart, WP Carey and State Street" on September 27, 2016.
CUBE Company Profile
CubeSmart, incorporated on July 26, 2004, operates as a self-managed and self-administered real estate investment trust (REIT). The Company's activities are conducted solely through CubeSmart, L.P. (the Operating Partnership) and its subsidiaries. The Firm is focused primarily on the ownership, operation, management, acquisition and development of self-storage facilities in the United States. The Company's self-storage facilities offer storage space for its residential and commercial clients with features, such as wide aisles and loadbearing capabilities for large truck access. The Company's self-storage facilities are located in metropolitan areas, as well as suburban areas. The Firm owns over 440 self-storage facilities located in over 20 states and in the District of Columbia containing an aggregate of approximately 30.4 million rentable square feet. It also enables the clients to rent storage cubes for their use, typically on a month-to-month basis.
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Bring on the autumn! There's not much better to lift your spirits than jumping through a big pile of crunchy leaves with your kids. While you're out and about enjoying the new season, collect up some leaves and bring them home to play these seasonal letter and maths games.
Homemade paint recipe with spices
This homemade paint recipe with spices is as much a delight for the nose as the eyes! It's super easy to make and great for both sunny days when you're painting outdoors, or dark days when your senses need a boost. Here's how to make it.
Homemade paint recipe with spices [Read more…]
Autumn art from The Garden Classroom
We've been combining two activities from my book The Garden Classroom this week to create some beautiful autumn art. Taking the idea of leaf art and adding it to my Sticky Picture frames, here's what we made. [Read more…]
Nature walk ideas :: Simple Play
Welcome to day nine of our Simple Play :: complex learning series, where today we're sharing nature walk ideas. Read on for ideas for observing the seasons and using your walk as a springboard into art, craft and sensory play.
Nature walk ideas [Read more…]
20 school garden ideas for autumn and winter
Just because the days are getting cooler and shorter doesn't mean children have to stop enjoying lots of outdoor play. Having fun and creative garden projects on offer is a great way to keep kids active outdoors and enjoying the fresh air.
The school gardening club I run is back in action after the school holiday, with a group of young gardeners ready for action. Here are 20 ideas you can use over the autumn and winter terms – at home as well as in a school garden – to enjoy the outdoors all year round, with some special activities taken from my book The Garden Classroom.
20 school garden ideas for autumn and winter [Read more…]
Garden chalkboard puppets
Transform your block play with chalkboard paint!
Up! from blocks to rockets
Kids gardening activities : chalk
We're having so much fun exploring all the kids gardening activities we can come up with using chalk this week. Taking One Added Ingredient and introducing it to lots of different areas of play gets children thinking and imagining and creating. Using a material they are familiar with ~ in our case chalk ~ but adding it in different ways leads to all sorts of new play possibilities. On Monday I showed you how we'd use chalkboard paint to make some ever changing puppets, in our fairy garden and in a road map right across the patio. Today we're using chalk in a treasure hunt that mixes in lots of language, maths and nature play too.
Kids gardening activities : chalk treasure hunt [Read more…]
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Enter ePHOTOzine's Prize Draw, with fab gifts for everyone! Click Here
Love the play of light on the Bracken and the manner in which it slants through the trees. I have indeed visited this lovely forest and recall its many beauties well. Hope you are well? We have had close on 2 weeks of glorious sunshine up here. Not too sure what we've done to deserve it!
Aww, thank you Ann, the light that morning was gorgeous We are lucky that we can visit whenever we want to be honest, but unfortunately due to pressures of work/distance we don't get there as much as we would like. It is indeed a beautiful place to visit, preferably out of season I am very well thank you....currently looking for another job, not because I have to, but need to move on Hope all well with you
Julie xx
Julie xx
Lovely bit of light on the bracken, Julie. Don't start the countdown to Christmas too soon
What a beautiful shot Julie,superb lighting hitting the bracken..Lin
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With our Live A Life Of Love silicone bracelet set you can tell the world that God loves you and is a God of love. Each bracelet is printed all the way around.
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For Sale!
Wanted: Buddhist Neighbor for Shambhala King.
Just got this notice via my longtime Buddhist pal Mike Smith, and thought I'd pass along—it's not every day you get a chance (even if you have the dough) to live next to the King of Shambhala. Maybe you could rake leaves together, shoot the shizzle about the weather! Bring over your favorite green tea! Trade tips on comfortable gomdens!
But be warned: you must be okay with lhasangs, occasional Tibetan dancing, and lots and lots of peaceful meditation.
Hey Way,
The house next door to the Sakyong's that is for sale, for $1.6 million CDN bucks.
Some of the Sakyong's other neighbors that are Sangha Members have expressed hope to me that we find a sangha member to buy the property. I'm sure we could negotiate the price.
If you know someone who might be interested in placing an offer on this property please call me—my contact information is below.
Here's a link for the details of the property. Excerpt:
Waterfront Paradise on the shores of the Northwest Arm. Extensively renovated with all of today's modern finishes, this estate property has it all including detached nanny/in-law suite. 5 bedrooms, 3 full baths, 1.5 garage, quality custom finishes…
Yours In the Vision,
Mike Smith: | T: 902.209.6745 | W: www.iListinHD.com
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Halifax, Nova Scotia Real Estate Blog Beautiful Real Estate on the Halifax Peninsula
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September 23, 2011
EDWARD GRIMES, ET AL., PLAINTIFFS,v.CORRECTIONAL MEDICAL SERVICES, ET AL., DEFENDANTS.
Presently before the Court is a motion brought by Plaintiffs pro se Edward Grimes, John Tell, and William Engel ("Plaintiffs"), seeking leave of the Court to amend the Complaint. Docket Entry No. 129. The proposed Amended Complaint (the "2011 PAC") would add 14 new defendants, several new claims including violation of the federal Racketeer Influenced and Corrupt Organizations Act, 18 U.S.C. § 1961, et seq. ("RICO"), violation of the New Jersey RICO statute, N.J.S.A. 2C:41-1, et seq., violation of public policy regarding the medical care of state prison inmates, violation of the New Jersey Constitution, violation of the Fourteen Amendment of the United States Constitution, and breach of contract against proposed new defendant, University of Medicine and Dentistry of New Jersey ("UMDNJ"), as well as a request for additional relief including a preliminary injunction against all Defendants. Plaintiffs also seek to add new factual allegations and theories of liability. Defendants, Correctional Medical Services, Inc. ("CMS"), Joseph Girone, M.D., Allan Martin, M.D., Ihuoma Nwachukwu, M.D., Grace Melendez, M.D., Donique Ivery, N.P., Jawana Bethea and Jason Pugh, (collectively, the "CMS Defendants") oppose the motion on the grounds of undue delay and accompanying bad faith in that delay, undue prejudice, and futility. See CMS Brief in Opposition ("CMS Opp.") at Docket Entry No. 138; CMS Sur-Reply ("CMS Sur-Reply") at Docket Entry No. 145; CMS Supplemental Brief in Opposition ("CMS Supp. Opp.") at Docket Entry No. 147. Defendants George Hayman, as Commissioner of the Department of Corrections (the "DOC") and Michelle Ricci, as Administrator of the Prisons (collectively, the "DOC Defendants"), also oppose the motion on the grounds that the motion is untimely, unduly prejudicial, and futile. See DOC's Brief in Opposition ("DOC Opp.") at Docket Entry No. 140; DOC Supplemental Brief in Opposition ("DOC Supp. Opp.") at Docket Entry No. 146. For the reasons stated below, Plaintiffs' motion is DENIED.
I. Statement of Facts and Procedural History
A. The Original Complaint
Plaintiffs filed their original Complaint on or about February 1, 2008 (the "Complaint"), setting forth allegations that Defendants*fn1 deprived Plaintiffs, as inmates of the New Jersey State Prison ("NJSP"), of vital medications, and caused them severe pain, lasting negative health consequences, and emotional harm.
Plaintiffs state that the DOC contracted with CMS, a private medical services provider, to provide medical care to inmates incarcerated in NJSP. Complaint ¶¶ 64-65. Plaintiffs suffer from a multitude of health issues, which require frequent interaction with the prison medical system. See Complaint ¶¶ 68-77, 108-128, 199-201. Each Plaintiff suffers from severe heart problems in addition to several other illnesses, and Plaintiffs describe in the Complaint the many medications they must each take to control these health conditions. Id. Plaintiffs maintain that their complicated medical histories make it necessary to keep their medication regimens as prescribed by their specialists, and that any lapse in the dosage will cause them harm and could potentially lead to death. See Complaint ¶ 238.
Plaintiffs allege that CMS and its employees: 1) failed to adequately provide Plaintiffs with their prescribed medications; 2) altered or ignored prescriptions and advice of specialists; and 3) retaliated against Plaintiffs for disagreeing with the staff or treatment given. See, e.g., Complaint ¶ ¶ 86-96, 142, 147, 164, 204-205, 224-225. Furthermore, Plaintiffs claim their administrative remedy forms to both CMS and the DOC have either gone unanswered or been returned unaddressed, and that the DOC Defendants have failed to adequately ensure that proper medical services were being provided. See, e.g., Complaint ¶ ¶ 12-41.
Plaintiffs allege that Defendants, through their actions and inactions, have violated Plaintiffs' Eighth Amendment right to be free from cruel and unusual punishment. Additionally, Plaintiffs claim CMS violated its contract with the DOC to provide adequate medical services, and Plaintiffs assert that they were the intended beneficiaries of that contract. Finally, Plaintiffs seek damages and relief based on negligence and intentional infliction of emotional distress. Plaintiffs pray for compensatory and punitive damages, and a permanent injunction which orders Defendants to "1) immediately comply with all recommendation by medical specialists for a treatment plan with (sic) includes pain management; 2) keep all prescriptions up to date; 3) find the present remedy system null and void, and implement a system where the prisoner retains a copy of any remedy filed for proof of filing; 4) waive all fees for medical treatment for chronic care; and, 5) Allow patient to refuse placement in the infirmary." Complaint ¶ 240.
On May 2, 2008, the Court proceeded the Complaint after it was screened in accordance with 28 U.S.C. § 1915. Docket Entry No. 5. Prior to this date, the CMS Defendants filed a motion to sever the case, requesting that each Plaintiff prosecute his own action. Docket Entry No. 3. On July 7, 2008, Plaintiffs filed a brief in opposition to the motion to sever and a cross-motion to amend the complaint and appoint pro bono counsel. Docket Entry No. 7.
B. The 2008 Proposed Amended Complaint
In their first proposed Amended Complaint (the "2008 PAC"), Plaintiffs sought to add a significant number of new defendants, causes of action, and theories of liability, as well as an additional named plaintiff, with whom they sought class certification on behalf of all current and released inmates of NJSP who had been denied adequate medical treatment.
Two of the new causes of action in the 2008 PAC asserted violations of the federal and state RICO statutes. Plaintiffs claimed that the medical staff refused to disburse medications and then falsified Plaintiffs' medical records by marking the files to reflect that the medications were distributed. See, e.g., 2008 PAC ¶ 70. According to Plaintiffs, CMS Defendants engaged in wire and mail fraud when the falsified medical records were transmitted to CMS headquarters and other CMS centers out of state. See 2008 PAC ¶ ¶ 195-198. Plaintiffs contended in the 2008 PAC that the CMS Defendants omitted information in their medical records from their specialist physicians or otherwise altered Plaintiffs' records in order to funnel prescriptions through the CMS-owned pharmacy. See 2008 PAC ¶ ¶ 67-71.
In addition to the RICO claims, Plaintiffs sought to add new causes of action and new theories of liability for violation of their Fourteenth Amendment rights, medical malpractice, violation of the New Jersey Constitution, and violation of several state and federal statutes which Plaintiffs argued represented the public policy with regard to providing medical care in state prisons. Plaintiffs claimed that not only did the CMS Defendants fail to provide necessary medical services and engage in racketeering activity, but that the DOC Defendants knew about the CMS Defendants' activity and failed to enact policies to rectify and prevent these wrongs against Plaintiffs. 2008 PAC ¶¶ 71, 75-87.
Finally, Plaintiffs sought to expand their prayer for relief in the 2008 PAC to request the appointment of a special master to preside over the DOC medical programs, and that the Court grant a preliminary injunction in addition to the permanent injunction previously sought, with the additional conditions of requiring Defendants to provide Tell and Engel with their requested medical aids (such as special chairs and mattresses), to refrain from retaliating against Plaintiffs, to provide Plaintiffs with medications as prescribed by their doctors, and to utilize local pharmacies if the medications could not be timely filled through the CMS pharmacy. 2008 PAC
¶¶ 228-232. Plaintiffs contended that in April of 2008, the DOC signed a new contract with UMDNJ for the provision of all medical services in NJSP, but that UMDNJ would not fully take over until September 2008. 2008 PAC ¶ 54. As UMDNJ had yet to perform under the new contract, Plaintiffs sought only declaratory and prospective relief against UMDNJ. 2008 PAC ¶ 45.
The Court reviewed Defendants' motion to sever in conjunction with Plaintiffs' cross-motion to amend. In an Order dated December 19, 2008, the Court found that requiring each of the three Plaintiffs to proceed separately would "neither assist in the management of the docket nor serve the interests of judicial economy," and therefore denied the CMS Defendants' motion to sever. Docket Entry No. 36. Furthermore, in noting that Plaintiffs' cross-motion depended in part on the outcome of the motion to sever, the Court denied Plaintiffs' cross-motion to amend without prejudice, and expressly granted them leave "to file new motions seeking any relief sought in the cross-motion as may still be appropriate," in light of the denial of the motion to sever. Id. In a footnote, the Court further clarified that the "Plaintiffs should file a separate motion for each type of relief they seek." Id.
C. The 2011 Proposed Amended Complaint
Following the Court's ruling on the various motions, the parties engaged in discovery. The parties also conducted extensive motion practice, with the CMS Defendants filing motions for partial summary judgment and Plaintiffs filing motions to compel discovery and for preliminary injunctions. See Docket Entry Nos. 47, 55-57, 70, 80 and 82. Due in part to the motion practice and the considerable discovery which was being exchanged, at the request of both sides, the Court entered several Orders extending the time for completion of fact discovery. See Docket Entry Nos. 51, 67, 91, and 106. The October 20, 2010 Scheduling Order provided for all fact discovery to be completed by December 1, 2010 and for dispositive motions to be filed no later than December 23, 2010. Docket Entry No. 106. The deadline for the filing of dispositive motions was extended to April 8, 2011 by Order dated February 25, 2011. Docket Entry No. 127. On February 28, 2011, in all likelihood before Plaintiffs received the Order extending the dispositive motions deadline, Plaintiffs filed both a motion for summary judgment and the instant motion for leave to file the 2011 PAC.
On March 10, 2011, the Court conducted a telephone conference to discuss the timing of the simultaneous motions for summary judgment and leave to amend. After a discussion on the record, and in consideration of the resources of all the parties and judicial economy, the Court terminated the motion for summary judgment pending resolution of the motion to amend. See Docket Entry No. 134, Order memorializing the conference.
Plaintiffs' 2011 PAC seeks to add 14 new defendants, five new causes of action, new factual allegations, and several new theories of liability.*fn2 Much of 2011 PAC overlaps with claims and facts alleged in the 2008 PAC. For example, of the 14 new defendants, seven were named in the proposed 2008 PAC, and all of the causes of action Plaintiffs seek to add now were alleged in the 2008 PAC. Some causes of action, however, have been extended to incorporate new defendants and update allegations. For example, Count II Breach of Contract is now alleged against CMS and UMDNJ, as UMDNJ took over medical services in October 2008. See 2011 PAC ¶ 293. Plaintiffs' prayer for relief is almost identical to that in 2008, including the request for preliminary and permanent injunctions. In addition, many of the theories of liability are substantially similar if not identical to those alleged in the 2008 PAC.
II. Analysis
A party may amend its pleading as a matter of course within 21 days of service, or if the pleading is one to which a responsive pleading is required, the party may amend as a matter of course within 21 days after the service of a responsive pleading or 21 days after service of a motion under Rule 12(b), (e), or (f), whichever is earlier. Fed. R. Civ. P. 15(a)(1). After such time, a party must seek the written consent of the opposing party or leave of court. Fed. R. Civ. P. 15(a)(2). Under Rule 15(a)(2), leave should be freely given, and the Third Circuit has taken a liberal approach on allowing amendments under this rule so that claims may be decided on the merits and not any technicality. Fed. R. Civ. P. 15(a)(2); WHY ASAP, LLC v. Compact Power, 461 F.Supp.2d 308, 311 (D.N.J. 2006). Furthermore, as Plaintiffs are pro se, the Court must be mindful to construe the pleadings liberally in favor of Plaintiffs and not hold them to the stringent standards of formal pleadings drafted by lawyers. Erickson v. Pardus, 551 U.S. 89, 94 (2007). Absent "undue delay, bad faith, dilatory motive, unfair prejudice, or futility of amendment," the court should grant leave to amend. WHY ASAP, 461 F.Supp.2d at 311.
A. Undue Delay
Delay alone is insufficient grounds to deny an amendment. Cornell and Co., Inc. v. Occupational Safety and Health Review Commission, 573 F.2d 820 (3d Cir. 2001). However, "at some point, the delay will become 'undue,' placing an unwarranted burden on the court, or will become 'prejudicial,' placing an unfair burden on the opposing party." Adams v. Gould, 739 F.2d 858 (3d Cir. 1984). There is no calendar that marks an amendment's aging from timely to delayed, nor a date at which delay become "undue." See Chisler v. Johnston, 2011 WL 3209092 *3 (W.D. Pa. July 28, 2011). Rather, the court focuses its inquiry on the motives or reasons the moving party failed to amend earlier. Adams, 739 F.2d at 868. A court may deny a motion to amend if the moving party cannot satisfactorily explain its delay. Harrison Beverage Co. v. Dribeck Importers, Inc., 133 F.R.D. 463, 468 (D.N.J. 1990), citing Leased Optical Dept. v. Opti-Center, Inc., 120 F.R.D. 476, 478 (D.N.J. 1988).
Defendants argue undue delay because the record shows that Plaintiffs could have sought to file the proposed amendments earlier. The baseline for Defendants' opposition is that "the claims which Plaintiffs in 2008 sought to add to this litigation were either similar or identical to the claims which Plaintiffs now seek to add." CMS Opp. at 12. Defendants contend that the similarities between the 2008 and 2011 PACs demonstrate that Plaintiffs had the necessary information to file an amended complaint at an earlier point in the litigation. CMS Opp. at 13. Defendants further point to Plaintiffs' numerous other filings to show that Plaintiffs had ample opportunity and ability to file the motion. CMS Opp. at 12-14; DOC Opp. at 16.
Plaintiffs allege the delay in filing the 2011 PAC was caused by Defendants themselves, because of what Plaintiffs perceive as Defendants' "unwillingness to answer pro se discovery request[s]." Plaintiffs' Reply ("Plaints' Reply") at 4. Plaintiffs describe the various letters and exchanges from June 2009 through June 2010, which include a motion to compel in September 2009, receipt of an 8,000 page document production in September 2009, a second motion to compel in April 2010, and a reformulation of Plaintiffs' interrogatory requests in June 2010. Plaints' Reply at 2-4. Plaintiffs claim they particularly sought the names of those individuals who withheld or delayed their medications and Plaintiffs specifically cite their request for the work schedules of those nurses who disbursed medications in the unit in which Tell and Engel lived. Plaints' Reply at 3-4. Plaintiffs state they were waiting for this information to file their amended complaint. See Plaints' Reply at 4, citing Exhibit 1, Docket Entry No. 143-1, at 68.
Defendants contend that they have produced thousands of pages of medical records and that the discovery at issue, even if delayed, could not form the basis for the proposed amendment. DOC Opp. at 17-18. By way of example, the DOC Defendants cite the specific nurses' shift schedules referenced by Plaintiffs; while Defendants say they have produced many of the other shift records for both CMS and for UMDNJ, the DOC and NJSP did not have those particular records in their custody to produce. Id.
Plaintiffs offer two other reasons for delay. Tell argues that he was carrying the bulk of the prosecutorial burden and was suffering from health complications that slowed and affected his ability to prosecute the case. Plaints' Reply at 5. Plaintiffs also state they were under the mistaken belief they had until after dispositive motions to amend the complaint. Id.
The Court finds undue delay in Plaintiffs' motion to file the 2011 PAC at this late date in the litigation. Plaintiffs have not explained why they waited more than two years to re-file what are predominantly the same claims and facts asserted in the 2008 PAC. The vast majority of the new allegations are dates and details about when medications were or were not given and interactions with various medical service employees. Essentially, most of the new facts are updates of information in the original Complaint and the 2008 PAC, but not new allegations. Furthermore, these new facts consist of information that was within the control of Plaintiffs and did not require discovery to uncover it. In light of this, the Court finds that the discovery issues allegedly caused by Defendants' resistence in production of documents are insufficient to explain the delay.
As for Plaintiffs' additional reasons for delay, while the Court is sympathetic to the health issues that Plaintiffs suffer, the Court cannot excuse a more than two year delay to amend the Complaint when put in context of Plaintiffs' steady motion practice. Several major motions and declarations have been filed by Plaintiffs since the December 19, 2008 Order. Plaintiffs claimed they have made references to a future amendment, but such references are not enough to put Defendants and non-parties on notice of potential claims which may be leveled against them.
B. Unfair Prejudice
Prejudice to the non-moving party "is the touchstone for the denial of an amendment," and a finding of unfair or substantial prejudice is sufficient ground for denial. Cornell and Co., Inc., 573 F.2d at 823; Lorez v. CSX Corporation, 1 F.3d 1406, 1414 (3d Cir. 1993). A finding of unfair prejudice may be made where the non-moving party would be "disadvantaged or deprived of the opportunity to present facts or evidence it would have offered." Harrison Beverage Co., 133 F.R.D. at 468. In evaluating the extent of any alleged prejudice, the court looks to the hardship on the non-moving party if the amendment were granted. Cureton v. NCAA, 252 F.3d 267, 273 (3d Cir. 2001). "Specifically, [courts] have considered whether allowing an amendment would result in additional discovery, cost, and preparation to defend against new facts or theories." Cureton, 252 F.3d at 273.
Defendants argue that Plaintiffs' amendments will prejudice them because of additional discovery that will need to be done, resulting in delays and heightened costs. Defendants cite to the breadth of the new amendments, especially the RICO claims and Plaintiffs' other new theories of liability, and assert that if the amendments were allowed, it "would require a full-scale reopening of discovery." DOC Opp. at 18-19; CMS Opp. at 15. They point to the fact that not only would each new defendant be entitled to conduct discovery on the claims, but each would also be entitled to revisit the discovery already exchanged among the current parties. CMS Opp. at 15.
Moreover, the DOC Defendants argue prejudice as to the new parties that Plaintiffs seek to add, given that this case has been pending for over three years, and these proposed new parties have not had notice of the claims or an opportunity to prepare a defense. The DOC Defendants conclude that "[m]ost if not all new proposed defendants have had no notice of or reason to know of this action, and as to existing defendants, the new theories of liability, and thus no reason to assume they were mistakenly omitted from it three years ago." DOC Opp. at 19.
Plaintiffs respond that Defendants cannot claim to be surprised by the proposed amendments. Plaintiffs point to both the 2008 PAC and their references over the years to their intention to amend. Plaints' Reply at 5. Thus, Plaintiffs argue they put Defendants on notice of the potential claims. Id.
The Court finds that it would be prejudicial to both the current Defendants and to the new Defendants to allow the 2011 PAC to proceed. Plaintiffs assert several new, unrelated, and complex claims against the current Defendants and also seek to add 14 new Defendants not previously named in this suit. The current Defendants would need to engage in further protracted discovery regarding the new claims, and the new Defendants would have to conduct their own discovery for any claims asserted against them, in addition to accessing all discovery already exchanged among the parties. Parties who have already been deposed would likely need to be re-disposed at substantial additional expense. Discovery in this case has already been an extended and contentious process. Beginning in March 2009, the schedule was extended multiple times, and fact discovery finally closed in December 2010. See Amended Scheduling Orders, at Docket Entry Nos. 44, 51, 67, 79, 91, and 106.
Moreover, when the instant motion was filed, this case was on the threshold of dispositive motions. See Docket Entry No. 127. In fact, Plaintiffs filed their dispositive motion on the same day as their motion to amend, but the Court, after a discussion with the parties, terminated that motion pending the outcome of the motion to amend. See Docket Entry No. 134. Re-opening discovery at this point and to the scale required by the 2011 PAC would in effect reset this case to its inception.
The Court finds that in addition to the prejudice which stems from the cost of discovery, Defendants would be further prejudiced by the delay of the 2011 PAC in their ability to pursue their defenses. This effect is exemplified in Cornell and Co., Inc., 573 F.2d 820, where the moving party, the Occupational Safety and Health Review Commission, sought to amend the Complaint before the administrative law judge to change the underlying theories of liability, several months after the Commission cited Cornell and Co. for violating safety regulations. Id. at 824. The court, in overturning the administrative law judge's grant of leave to amend, found the passage of time had severely eroded Cornell and Co.'s ability to call witnesses and find evidence to defend against the newly added theories. Id. The same holds true here. The majority of the factual allegations refer to incidents in 2006 to 2008. As mentioned by the DOC Defendants, none of the Defendants had reason to believe these new claims would become part of the case after such a long passage of time. Requiring Defendants to begin collecting evidence now to defend against new claims and factual allegations that occurred three to five years ago presents a substantial burden.
The mere fact that Plaintiffs alleged similar if not identical claims in 2008 did not serve to put Defendants on notice or give them an opportunity to prevent prejudice. Since 2008, this case has proceeded on the claims set forth in the Complaint. Discovery has been tailored to these allegations, not the ones alleged in the 2008 PAC. Furthermore, contrary to Plaintiffs' belief, the 2008 motion to amend did not put the proposed new defendants on notice. Nothing in the record reflects that Plaintiffs served each potential new Defendant with a copy of the 2008 PAC, and thus, unless otherwise notified, the new Defendants would have had no knowledge of the claims laid against them in 2008. Accordingly, this Court concludes that allowing the amendments would prejudice Defendants.
C. Bad Faith
The CMS Defendants argue that Plaintiffs acted in bad faith in delaying two years before seeking to file the 2011 PAC. Courts looks to the motives of the moving party to determine bad faith. Adams, 739 F.2d at 868. The CMS Defendants make no more than conclusory statements that a two year delay evidences bad faith. Nothing in the record suggests that Plaintiffs operated in bad faith, and the Court therefore declines to find Plaintiffs have acted in bad faith, despite the delay.
D. Futility
Defendants argue that several of Plaintiffs' claims are futile. Defendants contend that Plaintiffs failed to allege facts which if true would constitute a violation of RICO. See DOC Opp. at 21; DOC Supp. Opp. at 7-10; CMS Supp. Opp. at 9-10. The DOC Defendants also argue that Plaintiffs have no standing to assert claims for third-party benefits under the service contracts with CMS or UMDNJ, nor is there a private cause of action to assert the violations of public policies or the attendant statutory provisions which Plaintiffs cite. DOC Opp. at 22.
The Court determines it need not reach the merits of the futility argument, given its conclusion that the proposed amendments should be denied due to both undue delay and unfair prejudice. For these reasons, as more fully set out above, the Court denies Plaintiffs' motion for leave to amend.
III. Conclusion
In conclusion, the Court finds that Plaintiffs have not sufficiently explained the lengthy delay in filing a motion for leave to amend the Complaint, given that Plaintiffs were clearly aware of the claims and the identities of proposed new Defendants that they seek to add now, as early as 2008 as evidenced by the 2008 PAC. Furthermore, the breadth of new claims and the sheer number of new Defendants would require that discovery not only be re-opened but largely repeated, many months after it was closed. Accordingly, the Court finds the Plaintiffs' request for leave to amend must be DENIED.
ORDERED that Plaintiffs' motion for leave to amend, Docket Entry No. 129, is hereby DENIED; and it is further
ORDERED that as set forth in this Court's March 24, 2011 Order, Docket Entry No. 134, Plaintiffs are granted leave to re-file their motion for summary judgment upon letter request without having to physically resubmit the papers, or if they choose to modify the motion, they may do so. In either event, any dispositive motions by either Plaintiffs or Defendants are to be filed by no later than November 23, 2011; opposition is to be filed by no later than December 12, 2011; reply is to be filed by no later than December 27, 2011; and, the motion is to be returnable on January 3, 2012. There will be no oral argument on the motions unless specifically noticed by the Court.
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- Customized Hardscape in Savage, MN
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Manufacturing Area
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Posada Los Mangos,Isla de Margarita.
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Free personalized radio that
plays the music you love
Features of This Trackhouse roots
four-on-the-floor beats
disco influences
danceable beats
a rhythmic intro
epic buildup/breakdown
use of modal harmonies
light drum fills
synth swoops
effected synths
ambient synth textures
trippy soundscapes
prevalent use of groove
These are just a few of the hundreds of attributes cataloged for this track by the Music Genome Project.show more
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text:
Red V-neck Top With Lace At Shoulders
A red colour v-neck top for women. The top features batwing sleeves with lace inserts and a stylish loose fit. Style the top with ecru slim fit trousers and red heels for a stunning style statement.Fabric: 100% Cotton
Fit: Relaxed
Model Statistics: Height - 178 cm / 5 ft. 10 inches.
                            Bust - 81 cm / 32 inches.
                            Waist - 61 cm / 24 inches.
Model is wearing size XS.
Care Instructions
Which size will fit me?
you'll fit right into a...


text:
Min comment interval: 14 seconds
Remaining character count: 4000
Remaining character count: 4000
Spoiler Image
Shortcuts: "C" opens comments. "R" refreshes comments.
Record voice message?
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> hey anon, wanna give your opinion?
2 new threads. 1 new replies.
#24 - redyoshi
Reply +10 123456789123345869(03/24/2013) [-]
It's not the liquid that's science, the white coating around the liquid repels the liquid, making it so the liquid can't go past the coating. The spray is called NeverWet
#32 to #24 - largeheadphones
Reply +3 123456789123345869(03/24/2013) [-]
The spray is called your **** will never get wet.


text:
The D38999/20WG16BD from Deutsch is MIL-DTL-38999 Series III wall mount receptacle with 16 contacts. MIL-DTL-38999 Series III connectors are available in wide range of shell materials and finishes with 48, 500, 1000 and 2000 hour salt spray plating exposure. These connectors offer high density contact arrangements in a miniature circular shell and combines high coupling durability. It is designed to withstand the extreme shock, exposure and vibration.
Conn Circular SKT 16 POS ST Flange Mount 16 Terminal 1 Port


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Stuck on your essay?
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1 Page
Why Early Marriages are Wrong and Harmful
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Pages: 1
Word Count: 375
Category: Sociology
Word Count: 375
Pages: 1
In the Middle East many years ago people used to get married at an early age For example the husband used to get married at the age of sixteen and the wife used to get married at the age of thirteen this custom is not used a lot nowadays as families used to do in the past Although some people still get married at an early age I think early marriage is harmful for husband and wife Some people still think that getting married at an early age is better for several reasons Their first reason its a tradition and they have to follow their traditions so they get married at an early age and it has been successful for many years The second reason chastity which let them marry their children of to preserve their children from going astray that is against their cultures rules The third reason they believe that getting married at an early age can make their children learn how to be responsible However these reasons are not very valid as for the first reason because time is different and there are many traditions that have been changed because children started to think on a different way Regarding the second reason that is not very valid because these days girls and boys are more aware than the past For the third reason kids nowadays can learn how to be responsible without getting married because of the new ways that can teach children how to be responsibleIn my opinion getting married at an early age is harmful because of several reasons The first reason is that kids at the age of sixteen are not immature enough for getting married and they have to be leaded by someone the second reason children at this age have no jobs opportunities because they have to have time to get educated The last reason children
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text:
Description from the manager
Description from the manager
Bedrooms: 3/
Bathrooms: 3/
Sleeps: 8/
Type: Villa
Book this 3 bedroom, 3 bathroom villa in La Cruz de Huanacaxtle today! A total of 8 people can sleep here comfortably.
Guests can enjoy a pool, air conditioning, and parking space among other amenities offered by this rental.
No travelers have shared a review of this property yet.
The owner responds to 70% of inquiries, which is lower than average, and last updated the property's calendar 2 months ago, so this rental may not be available for your requested dates, but please inquire to confirm.
About the manager
About the manager
Destino Vacation Rentals
Destino Vacation Rentals
- Response rate: 70%
- Response time: Within a day
- Calendar updated: 2 months ago
Overall rating:
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Proximity to TripAdvisor's Top La Cruz de Huanacaxtle Attractions
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Check in from 14h / Check out prior to 11h
Econo Lodge Inn & Suites Ft. Jackson: Find photos, reviews, information, amenities and compare prices for 2 stars hotels in Columbia, United States.
Econo Lodge Inn & Suites Ft. Jackson provides a comfortable setting while in Columbia. It also features a wake-up service.
Hotel description
The comfortable rooms at Econo Lodge Inn & Suites Fort Jackson Columbia include a TV, a fax machine and a coffee maker. They all come with a private bathroom that has a hair dryer.
A satisfying breakfast is prepared every morning at Econo Lodge, and there are numerous cafés and eateries nearby. There is also the added advantage of an on-site restaurant.
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text:
Never heard of Invisible Illness Awareness Week? You're not alone. Jody Smith sheds a little light to make it more visible
Discuss the article on the Forums.
It Seems Frightening At First, But This Is One Of The Most Beautiful Things I've Ever Seen.
Discussion in 'General ME/CFS News' started by Firestormm, Mar 6, 2014.


text:
This is the most ambitious project (fanart) that i've done -so proud- 🔥
Btw, it was a request from an anon "HT and GS going to grocery store (…)" so, thank you anon! TianShan headcanon on a store for you (and all) °ヮ° Yay ~
You can enter directly here to look full-> PAGE1 / PAGE2
Update:Thanks for all the love and support guys!
Oh! guys! i made a prequel! -> look!
Transformers Prime Bumblebee designs by Augusto Barranco.
Lionhead released some of the concept art for one of my Fable Legends characters, so I guess I can finally show it off! I haven't touched Glory since I handed her final orthos off to the team, so it's a blast to see her final model in all its glory. Pun very intended.
Glory's meant to be an Evil Queen archetype, but before she fully embraced the dark side- she's knows she's the shit and isn't very apologetic about it, apparently. Such a blast to design elegant adventure wear! CHOPINES FOR EVERYONE.
oh-so-casually I made this prequel "of my last comic" because i dunno @midotakaism you know why? Well "kiss on forehead" suggestion done °ヮ° for you guys all -proud 🔥-
Another Book of Life post! I had the fun challenge of figuring out how these soldiers worked in 3D space. Jorge originally designed the soldiers to function like a Picasso painting as a 2D asymmetrical silhouette. Even though their faces are always to camera, the modelers still needed the entire form figured out to help make their job of translating Jorge's 2D designs into 3D easier.
dancing through the dark
eerie shapes that cast the light.
between black shadows she sleeps,
waiting for their screams.
the goddess of magic and might,
the goddess of the moon and night,
call your ghosts upon those
who fear your power
over the heavens, the earth and the sea.
- hecate, goddess of witchcraft / e.a
So I've been working on this on and off the last couple days. An official turnaround for my often neglected baby, Nauta! I was gonna add some action poses & expressions, but I figure I'll be doing those down the line anyway.
Thinking of giving a certain villain this treatment too…
(Original Character Do Not Steal etc etc)
Metal Brick Waves
Higher quality versions: Ello | G+ | 2x@Dribbble | 960x960, nodes, & inspiration @ Imgur.
Wallpapers of this design are also available here.
Sometimes you fall off your horse.
Sometimes you look at that horse, and you say "Look horse, I'm going to ride you again. You see all those trophies we have on the wall? You know the legacy and the legend we leave behind? WE'RE NOT DONE YET." And then you get back on that horse, no matter how stubborn it is, no matter how difficult it makes it. And you ride it like you know what you're doing - because you have to, because you're as good as dead where you already stand. You have the experience to know that if you don't, you'll lose all relevance in the world's eyes as 5 others come to take your place.
And that's me getting back into the swing of motion graphics after the slump of creative productivity I've had this year. Things in life are DAMN good - I'm the happiest I've ever been, to be honest - but my creative productivity is down, and I really want for the graphic work that I do to flow naturally again. It currently feels like I'm fighting through an intellectual arthritis to get back into my own craft.
I also have the problem where I compare each of my new designs to the best pieces in my archives, and some of the newer designs just can't compare, so I'm demotivated to complete and post them. It's a struggle when I'm not comfortable with how my most recent work might be judged relative to my own previous works. It's double struggle when internally, I'm the one saying that this design isn't as good as a previous design. "Screw it", I have to say to myself. "The best pieces come from inspiration, momentum, and flow - and momentum is exactly what I need to build now." And so I will try to build momentum. Some of my upcoming designs may get simpler for a while while I build momentum. Let's hope that I can get back on this horse for another year, and make some beautiful designs again along the way. PS: I know nothing about horses.
This design's inspiration was sponsored by some painted brick walls I saw while driving home from work along a different path than normal due to a traffic jam.
Does anyone else have a hard time getting back on the horse after a few months out of whatever your flow is? How do you fight through that? Did you end up returning to, or surpassing your former glory when you got back into your craft?
Man, Ace! You can't even put down your cig for my orthographic references?
47. frustumation. can you see the pyramids?


text:
Find yourself on Flickr through tagging
Secretly its what most people do first when they come to a service like Flickr: they search for themselves. To see if anyone has uploaded a picture with you on it. For example the Flickr-feed of A4U Expo was without a doubt searched through by people looking for themselves (and they probably found Dave Naylor ;) ).
You can draw a face boundary on the photo, as you do with a note, and add their names. People in Photos has been wired into your your Recent Activity, so you'll always be up to date with who's added you to a photo or added other members to your photos.
For privacy reasons you can even set your own preferences, including who can add you to photos and who can add people to photos you've shared. And off course you can also remove yourself entirely if you want.
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The Federal NDP has turned their backs on many NDP values and shifted to the centre under the leadership of Thomas Mulcair.
A disturbing number of NDP member activists have been blocked from even seeking nominations.
What has happened to today's NDP federally and provincially was once unthinkable.
Is this . . . → Read More: Cowichan Conversations: Paul Manly Now Seeking Green Party Nomination in Nanaimo-Ladysmith


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2016 Panini Certified Football Hobby Box
10 packs per box. 5 cards per pack.
- Certified Football returns for 2016 with more stunning cards and stellar hits! Look for four autographs or memorabilia cards per box, on average!
- Look for iconic Freshman Fabrics Mirror Signatures, featuring supersized, short-print parallels with a larger memorabilia swatch!
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- 4 Autographs Or Memorabilia Cards, Including At Least One Rookie Patch Autograph Per Box On Average.


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Rieva Lesonsky, an AllBusiness.com writer, and former editor of Entrepreneur magazine started off her column recently with this:
The economy is in meltdown. Wall Street is floundering. Corporations are laying off thousands. The national unemployment rate is at a five-year high. But the New York Times thinks if you're an entrepreneur, you should close your business and get a job. Read.
She went on to share her feelings of anger at The NYT for only sharing stories of entrepreneurs who had to close down their businesses and were able to secure traditional employment successfully.
In the NYT article a couple of things do stand out:
1. There is no precise way to calculate how many people are closing their businesses and jumping to big companies.
2. THE unemployment rate rose to 6.1 percent in August. {A five year high}
We all know that the economic news has been lousy. We also know that the folks running for president are keeping the economy top of mind, as they should be, in their speeches. So does that mean that small business owners who are struggling should jump off a bridge?
I am a small business owner, myself. I own a small franchise consulting firm that was started in 1990 by my late father. I joined him in 2001. That was not a good year for several reasons. It certainly was not the best year to go into business.
2002 saw very high unemployment, but interest rates were pretty low, and the climate for business start-ups was getting a little better. My business is one that usually does better when unemployment is high, and hiring is light.
When there are lots of corporate downsizings, and not a lot of good jobs, more folks begin to think about non-traditional employment options. Some are able to leverage their most recent career tracks into freelance consulting projects that can either keep cash flow going until traditional employment options become more available, or can be the impetus for starting their own consulting firms.
There was a great deal of corporate downsizing in 2002-2004, and that did provide our firm with an above average amount of inquiries concerning franchise ownership options, and we were able to help several folks get into franchise businesses of their own.
Things are radically different now though…..
{Joel Libava is the President of Franchise Selection Specialists Inc. in Cleveland. Joel is a small business owner.}


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System Programming Analyst – Server Unix
Siam Commercial Bank Public Company Limited
Job Descriptions:
- Oversee day-to-day systems operation ensuring performance is in line with SLA targets
- Ensure the compliance with all security procedures across systems
- Develop and apply procedures for efficient systems installation, operation and administration
- Resolve significant system problems ensuring optimum system operation
- Provide development and testing environments for projects and enhancement activities
- Develop, apply control procedures and manage the migration of applications through the various technical environments
- Maintain, report system capacity response models and system capacity plans
- Communicate planned outage schedule and major issue status to Help Desk
- Coordinate with other IT Divisions in providing system maintenance and support
- Determine systems fail-over disaster recovery procedures and co-ordinate with IT Strategy and Architecture Planning Unit
- Bachelor's or Master's degree in Computer Science, Computer Engineering or any IT related fields
- 1-8 years of professional experience in areas such as Identity and Access
- Having experience on OS AIX or Linux platform by provision, install, configure, maintain and troubleshot AIX or Linux (Redhat) operating system
- Having skill on web – based application system by create, maintain and implement web-based application system and resolve issue and recommend enhancement if necessary.
- Knowledge of HTML, Java and related concept
- Able to work under high pressure
Interested candidate, please submit your CV via click ' APPLY NOW '
jobsDB จะส่ง ใบสมัครของคุณไปยัง Siam Commercial Bank Public Company Limited
Siam Commercial Bank Public Company Limited
"Let's start a brighter career future together"
Siam Commercial Bank (SCB) provides universal financial solutions to all customer segments with the best in benefits and full social responsibility. We are looking for responsible, enthusiastic, high caliber people with potential to join our team.
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คีย์เวิร์ด: System Programming Analyst


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The purpose of this study was to evaluate a commercial antimicrobial formulation, Byotrol™ G32, as a potential coating for impeding biofilm formation on medical devices such as urinary catheters.
Michelle Webb, School of Medicine, University of Manchester, Oxford Road,Manchester, M13 9PL, UK. E-mail: firstname.lastname@example.com
The purpose of this study was to evaluate a commercial antimicrobial formulation, Byotrol™ G32, as a potential coating for impeding biofilm formation on medical devices such as urinary catheters.
The antimicrobial activity of Byotrol™ G32 and its individual constituents has been tested on planktonic and biofilm cultures of uropathogenic bacteria. The Byotrol™ G32 formulation was superior with MICs ranging from 3 μg ml−1 to 15 μg ml−1 for planktonic cultures and 3–20 μg ml−1 for biofilms. Furthermore, Byotrol™ G32 was able to remove established biofilms and act as an antibiofilm surface coating.
Byotrol™ G32 displays impressive antimicrobial activity both in suspension and as a coating. Pretreating medical devices with Byotrol™ G32 may significantly impede biofilm formation and prolong the lifetime of the device.
Medical devices are indispensable in health care. They are, however, a predisposing factor in infection. This research has demonstrated that Byotrol™ G32 reduces bacterial growth and subsequent biofilm formation. Application of Byotrol™ G32 as a medical device coating could have a significant impact on the costs associated with device replacement and patient morbidity and mortality.
It is estimated that over half of all nosocomial infections are associated with medical devices, such as artificial heart valves, prosthetic devices, surgically implanted devices, contact lenses, wound drainage tubes, dressings, intrauterine contraception devices, sutures, intravenous catheters and urinary catheters (Smith et al. 1991; Costerton et al. 1999; Richards et al. 1999). Systemic and chronic infections are a result of biofilm formation on the surface of medical devices (Costerton et al. 1999). Biofilms are organized multicellular communities of bacteria attached to a surface and embedded in a protective polymer matrix. The biofilm phenotype is a ubiquitous characteristic of bacteria that constitutes a protected growth mode to facilitate survival in hostile environments (Costerton et al. 1987, 1995, 1999). It offers increased resistance to host defences and antimicrobials, and consequently, biofilms are notoriously difficult to treat and commonly manifest as chronic or recurrent infections (Patel 2005; Vuong et al. 2005; Anderson and O'Toole 2008). The most effective method to impede biofilm development is to avoid or reduce the initial adhesion of bacteria to the surface. The nonspecific attachment of bacteria to any surface is a key determinant in subsequent biofilm formation; therefore, many approaches have been adopted to prevent bacterial attachment to surfaces of medical devices (Banerjee et al. 2011; Knetsch and Koole 2011). Currently, the most widely used antimicrobial biomaterials are those that have silver-modified coatings (Boswald et al. 1999; Davenport and Keeley 2005). These are now routinely used in wound management (Davenport and Keeley 2005; Silver et al. 2006; Atiyeh et al. 2007), and while they can reduce the risk of infection, they introduce a host of problems. For example, the uptake of silver ions by bacterial cells has resulted in the emergence of silver-resistant strains (Silver 2003; Percival et al. 2005; Silver et al. 2006), indeed silver-resistant Pseudomonas aeruginosa has been isolated from burn patients who have been treated with silver-coated wound dressings (Modak and Fox 1981). Cationic compounds represent a suitable alternative as they define a structurally diverse class of antimicrobials (Banerjee et al. 2011). Such chemical diversity leads to a broad spectrum of activity and different modes of action. Furthermore, despite over a century of use, only trace levels of cross-resistance have been observed in the clinical environment (Gilbert and Moore 2005; Jaglic and Cervinkova 2012). Antibiofilm coatings containing novel cationic compounds could, therefore, have a significant impact on the clinical setting.
Byotrol™ G32 is a successful commercial antimicrobial hygiene product that is based on a novel proprietary mixture of poly(hexamethylene biguanide) chloride (PHMB); didecyldimethylammonium chloride (DDQ) and alkyl (C12, 70%; C14, 30%) dimethyl benzyl ammonium chloride (BAC). In this study, we evaluated the in vitro efficacy of Byotrol™ G32 for impeding biofilm formation.
The well-characterized laboratory strain Escherichia coli K12 (XL1 blue phage) and clinical isolates of E. coli, Klebsiella pneumoniae and Ps. aeruginosa obtained from urinary tract infections and characterized by Vitek® 2 (BioMérieux, Inc., Basingstoke, UK) were used in this study. The staff at the Central Manchester Foundation Trust, Clinical Sciences Building 2, Manchester, UK, kindly provided the clinical isolates used in this study. Stocks were stored at −80°C in 80% glycerol. Stock bacteria were cultured for 18 h on Luria–Bertani (LB) agar plates every two weeks. Plates were stored at 4°C. Overnight cultures were prepared by inoculating LB broth with several colonies from the working culture plates and incubating for 18 h with shaking at 200 rev min−1. The inoculum was standardized to 1 × 108 CFU ml−1 using the Miles and Misra method for the determination of viable cell counts. All cultures were incubated in an aerobic atmosphere at 37°C.
The composition of Byotrol™ G32 (Byotrol™ Technology Ltd., Daresbury, UK) is PHMB (poly(hexamethylene biguanide) chloride); DDQ (didecyldimethylammonium chloride) and BAC (alkyl (C12, 70%; C14, 30%) dimethyl benzyl ammonium chloride).
Poly(hexamethylene biguanide) was obtained from Arch Chemicals (Tradename-Vantocil TG) as a 20 wt% solution in water. DDQ was obtained from Lonza (Tradename - Lonza Bardac 2240) and received as a 40 wt% solution in water. BAC was obtained from Thor (Tradename – Acticide BAC 50 mol l−1) and received as a 50 wt% solution in water.
The minimum inhibitory concentration (MIC) of Byotrol™ G32 and its constituents was determined by incubating increasing concentrations of each with 100 μl of a 1 : 50 dilution of overnight cultures prepared in LB in 96-well flat-bottomed nontissue culture–treated polystyrene microtitre plates (Greiner Bio-one Ltd., Gloucestershire, UK). Eight technical replicates were prepared on each plate for each concentration of antimicrobial tested. The microtitre plates were incubated under aerobic, static conditions for 18 h at 37°C.
Positive growth controls were prepared by inoculating eight wells with 100 μl of 1 : 50 dilutions of bacteria and 100 μl sterile distilled water in which the antimicrobial was prepared. Negative controls were prepared by dispensing 100 μl LB broth and 100 μl sterile distilled water into a further 8 wells. The microtitre plates were incubated under aerobic, static conditions for 18 h at 37°C.
After the incubation period, the optical density at 595 nm (OD595) of planktonic growth was measured to quantify the MIC using a spectrophotometer (BMG Labtech FLUOstar OPTIMA). The average optical density (OD) from the eight negative control wells was subtracted from the average OD from the eight technical replicates of each concentration in the test wells. The MIC is determined as the lowest concentration of antimicrobial that completely inhibits visual bacterial growth, or an OD595 < 0·05.
After determining the MIC, the microtitre plate biofilm formation assay was used according to the method by Christensen et al. (1985). Briefly, excess media and any planktonic cells were decanted from the microtitre plate, and each well was washed with 200 μl sterile phosphate-buffered saline (PBS) (Sigma-Aldrich Company Ltd.). The plate was left in an inverted position to air dry overnight at room temperature. Each well was stained with 150 μl 0·4% (w/v) crystal violet (Sigma-Aldrich Company Ltd.) at room temperature for 10 min and washed with running tap water until the excess stain was removed and the running water appeared colourless. The plate was inverted and left to dry overnight at room temperature. The biofilm density was quantified by solubilizing the ammonium crystal violet stain with 200 μl 99·5% ethanol (Fisher Scientific UK Ltd., Loughborough, UK) and measuring the OD595 of solubilized crystal violet in each well using a spectrophotometer plate reader (BMG Labtech FLUOstar OPTIMA). The average OD from the eight negative control wells was subtracted from the average OD from the eight technical replicates of each concentration in the test wells.
Eight-well glass chamber slides (Lab-Tek™ Chamber slide™; Nunc, Fisher Scientific, Loughborough, UK) were used to analyse biofilms by bright field microscopy. Each slide had six increasing concentrations of antimicrobial agent, one in each well. The concentration of antimicrobial agent in each well was prepared by dispensing 150 μl of the antimicrobial agent and 150 μl of a 1 : 50 dilution of inoculum prepared from an overnight culture as described previously. The remaining two wells contained a positive growth control prepared with 150 μl inoculum and 150 μl water, and a negative control with 150 μl media and 150 μl water. Slides were prepared in triplicate. After incubation, the slides were washed with 1 ml PBS and immediately stained with 0·4% crystal violet for 10 min. Excess stain was washed with running water and left to dry in an inverted position overnight. The chambers were removed and microscopic images were collected on an Olympus BX51 upright microscope using a 100×/1·30 UPlanFLN objective. Images were captured using a CoolSnap HQ camera (Photometrics) through MetaVue Software (Molecular Devices). Images were then processed and analysed using ImageJ (http://rsb.info.nih.gov/ij).
This assay determines the effect of Byotrol™ G32 on biofilm cells. 200 μl of a 1 : 100 dilution of an overnight culture was dispensed into wells of a microtitre plate. The Minimum Biofilm Eradication Concentration (MBEC) assay is a modification of the method described by Ceri et al. (1999). A lid with protruding pegs (transferable solid-phase screening system, Nunc) was placed into the inoculated wells and incubated under static conditions for 18 h. The lid carrying pegs were transferred to a microtitre plate containing 200 μl sterile PBS, shaken to remove any nonadhered bacterial cells and then placed in a microtitre plate containing a concentration range of each antimicrobial, which was prepared in the same manner as described for the MIC assay and microtitre plate biofilm production assay. The plates were incubated under static conditions for 18 h. The pegs were then placed in 200 μl PBS, shaken briefly and immediately placed in 200 μl fresh sterile media and incubated for a further 18 h to allow for regrowth of viable bacterial cells on the pegs. After incubation, the OD595 of the plates containing any planktonic growth was read. The pegs were washed in 200 μl PBS and allowed to dry overnight at room temperature before placing the pegs in 200 μl 0·4% crystal violet for 15 min. The pegs were washed in running water and left to air dry overnight at room temperature. The biofilm density was quantified by solubilizing the ammonium crystal violet stain with 200 μl 99·5% ethanol and measuring the OD595 of solubilized crystal violet using a spectrophotometer plate reader. The average OD from the eight negative control wells was subtracted from the average OD from the eight technical replicates of each concentration in the test wells.
The wells of a glass chamber slide were pretreated by incubating overnight at 37°C with a 5 μg ml−1 and 1 mg ml−1 solution of Byotrol™ G32. After incubation, excess Byotrol™ G32 was decanted and slides were re-incubated for a further 18 h at 37°C to allow for evaporation of water. A 1 : 100 dilution of overnight cultures of bacteria was prepared in LB broth, 100 μl of which was added to pretreated slides and incubated for 8 h at 37°C. A minimum of four biological replicates were prepared for each concentration. After the incubation period, the slides were washed with 1 ml PBS, stained with 250 μl crystal violet and solubilized with 300 μl ethanol. The OD595 measurements were determined as for the microtitre plate biofilm formation assay.
Wells of a glass chamber slide were precoated with 300 μl of a 1 mg ml−1 solution of Byotrol™ G32 overnight at 37°C. After incubation, excess Byotrol™ was decanted and the slides were re-incubated for a further 18 h at 37°C to allow for evaporation of water.
Topographies were recorded using atomic force microscopy (AFM) (PSIA Inc, XE100, Surrey, UK) in noncontact mode. A commercial silicon cantilever (PSIA Inc, 910M-NSC15) with a nominal spring constant of about 40 N/m was used.
The glass slides with precoated Byotrol™ G32 were secured to a metal disc using double sided tape and installed on the AFM scanner. An area of 20 μm2 was scanned. A scratch was made across the bottom of the glass chamber using a 0·8 × 40 mm needle. The resulting topography image of this scratch gives an indication of the film thickness on the precoated glass slides.
The antimicrobial activity of the Byotrol™ G32 formulation and its individual constituents was tested on a planktonic laboratory E. coli K12 strain and clinical strains of E. coli, Kl. pneumoniae and Ps. aeruginosa, all isolated from patients with urinary tract infections. For the Byotrol™ G32 formulation, antimicrobial activity was observed against all isolates (Fig. 1) with MICs ranging from 3 μg ml−1 for the two E. coli isolates to 15 μg ml−1 for Kl. pneumoniae and Ps. aeruginosa. A comparison of the MICs for the individual constituents of Byotrol™ G32, displayed in Table 1, reveals that the G32 formulation outperforms the individual constituents.
|Minimum inhibitory concentration (μg ml−1)
|Byotrol™ G32
|E. coli K12
|E. coli clinical isolate
|Kl. pneumoniae clinical isolate
|Ps. aeruginosa clinical isolate
Next, the amount of biofilm produced by the remaining viable planktonic cells after 18 h was determined by crystal violet staining of the adhered bacterial cells. Figure 2 illustrates that low concentrations of Byotrol™ G32 (3–20 μg ml−1) reduce the biofilm development of all isolates. To confirm these results, biofilms stained with crystal violet were visualized by bright field microscopy (Fig. 3). In this instance, bright field microscopy of bacterial cells stained with crystal violet was preferable to fluorescence microscopy, as it avoided nonspecific binding of dyes to Byotrol™ G32, which we observed for the standard fluorescent biofilm labelling dye FilmTracer™ SYPRO® Ruby biofilm matrix stain (Invitrogen™, Life Technologies Ltd., Paisley, UK), and for the Live/Dead® stain (Invitrogen™). Figure 3 demonstrates that the number of adherent bacteria is substantially reduced in relation to the growth control, at concentrations of Byotrol™ G32 as low as 3 μg ml−1, confirming the ability of Byotrol™ G32 to impede biofilm development. Consistent with previous results, reducing Ps. aeruginosa biofilm development required higher concentrations of Byotrol™ G32. Furthermore, comparison to the minimum biofilm inhibitory concentrations for the individual constituents displayed in Table 2 reveals that again the G32 formulation outperforms any of the individual constituents in its ability to reduce the degree of bacterial growth.
|Minimum biofilm inhibitory concentration (μg ml−1)
|Byotrol™ G32
|E. coli K12
|E. coli clinical isolate
|Kl. pneumoniae clinical isolate
|Ps. aeruginosa clinical isolate
The impact that Byotrol™ G32 had on established biofilms was also determined by performing the MBEC assay. The results, which are presented in Fig. 4, show that Byotrol™ G32 was effective against biofilms of E. coli and Kl. pneumoniae; >50% of the biofilm was eradicated at concentrations of 60 μg ml−1. Ps. aeruginosa was more difficult to eradicate requiring 400 μg ml−1 to reduce the biofilm mass by 50%.
Byotrol™ G32 was used to coat glass surfaces by incubating a solution at 37°C and then drying. Atomic force microscopy was used to verify the extent of coating and access surface morphology. Figure 5(a) shows the atomic force micrographs of a glass surface coated with 5 μg ml−1 and 1 mg ml−1 Byotrol™ G32. A scratch drawn along the surface allowed the depth of the coatings to be measured (Fig. 5b). At a concentration of 5 μg ml−1, the coating depth was measured as 20 nm. At a concentration of 1 mg ml−1, the surface is evenly coated and the depth of the coating was measured as 60 nm (Fig. 5b). Biofilm development on the coated surfaces was determined by crystal violet staining of a biofilm grown for 8 h. On the surface coated with 1 mg ml−1 Byotrol™ G32, no biofilm development was observed for the two E. coli strains and Kl. pneumoniae, and Ps. aeruginosa biofilm development was impeded by ~60%.
Healthcare professionals, to support the care and treatment of patients, increasingly use medical devices such as catheters, shunts, orthopaedic implants and wound dressings. While medical devices offer regained structure and function to the body, they are a persistent source of infection (Darouiche 2001; Van and Michiels 2005). Such infections not only present profound economic burdens for society but infections associated with the insertion of a medical device lead to significant levels of morbidity and are sometimes fatal (Polonio et al. 2001). Bacterial communities known as biofilms play a central role in device-associated infections (Costerton 2007; Hatt and Rather 2008). Preventing biofilm formation is key to prolonging the lifetime of any medical device and reducing infection-related complications.
The ability of many conventional antimicrobials to inhibit biofilm formation has been assessed and some success has been found with the use of heavy metals, particularly silver. Hydrogel-coated latex catheters impregnated with silver on both the outer surface and lumen of the catheter have shown some effectiveness, however, only in the short-term (Bologna et al. 1999; Verleyen et al. 1999), possibly because the main challenge of the urinary catheter is the hugely mixed population of resistant micro-organisms that form a biofilm and cause infection, including bacteria, which display heavy metal resistance (Woods et al. 2009). Therefore, silver may not be an ideal inhibitor in the long term, as it may select for organisms with silver resistance. Furthermore, there is conflicting evidence in the literature as to the efficacy of silver in the clinical environment (Johnson et al. 2006).
Here, we have studied the antimicrobial and biofilm inhibitory properties of a proprietary hygiene product, Byotrol™ G32, which is a commercial formulation based upon a mixture of poly(hexylmethylbiguanide) chloride (PHMB), didecyldimethylammonium chloride (DDQ) and dimethyl benzyl ammonium chloride (BAC). PHMB is a polymeric cationic antimicrobial agent, which for many years has been used in the domestic, food and medical industries (Gilbert and Moore 2005; Kim et al. 2011). BAC and DDQ are quaternary ammonium compounds that have also been well studied as antimicrobial agents and used in a wide variety of settings (Ioannou et al. 2007). Although PHMB, DDQ and BAC all display impressive antimicrobial activity, we demonstrate that these constituents are synergistic in the Byotrol™ G32 formulation and present a much greater degree of antimicrobial activity.
The microtitre plate biofilm formation assay demonstrates a correlation between the MIC results for the inhibition of planktonic cells and the inhibition of biofilm forming cells, that is, the fewer the planktonic cells, the fewer the number of viable cells that are able to form a biofilm. However, the results from the microtitre plate biofilm formation assay validates the activity of Byotrol™ G32 against planktonic cells and therefore as an inhibitor of early biofilm formation. Ps. aeruginosa is inherently resistant to many antimicrobial agents due to surface factors such as its outer membrane impermeability and active drug efflux mechanisms (Drenkard 2003; Trott et al. 2007). The MIC for the planktonic cells of Ps. aeruginosa is lower than the biofilm inhibition concentration, which may be a result of fewer planktonic cells being killed; therefore, perhaps a greater number of cells were able to proliferate as a biofilm.
By comparison, Byotrol™ is least effective against Ps. aeruginosa. This may be explained by the suggestion that biocides, which have a broad spectrum of activity and nonspecific targets, may cause nonspecific resistance mechanisms, for example in Ps. aeruginosa, the hyperexpression of multidrug efflux pumps (Gilbert and McBain 2003). There may also be changes to the outer membrane, which reduces the permeability of the membrane to the biocide (McDonnell and Russell 1999; Gilbert 2005). However, the higher MIC of Byotrol™ for Ps. aeruginosa compared to that of the other organisms tested is likely to be due to increased tolerance rather than resistance (Gilbert and McBain 2003).
Intervention at the early stages of biofilm development and inhibiting planktonic cells before they are able to attach to a surface is most desirable. For this reason, the inhibition of viable planktonic bacteria to attach to the surface was evaluated at 18 h with the microtitre plate biofilm formation assay and 8 h for the pretreatment of a glass surface, as these time points reflect the early stages of biofilm formation.
Pretreatment of a surface with Byotrol™ G32 in this study provided the preliminary data as to the efficacy of Byotrol™ as an antimicrobial coating. AFM clearly shows the deposition of a nonstructured thin film over the concentration range for Byotrol™ G32. Glass has a negative zeta potential at pH 7 (Gu and Li 2000); therefore, we propose that as PHMB is a positively charged polyelectrolyte, it strongly adheres to the surface through opposite charge interaction (Borkovec and Papastavrou 2008) and serves to enhance the weaker adhesion of both BAQ and DDQ through the formation of a composite film.
To place these findings in a clinical context, the stability and lifetime of this coating under the flow of urine would also be an important factor in the success of Byotrol™ as a catheter coating. The more stable the coating, the longer the lifetime of the catheter. That being the case, any long-term toxicity associated with Byotrol™ against uroepithelial cells would also need to be assessed. It should be noted that PHMB and BAC, the compounds demonstrating the highest antimicrobial activity in the Byotrol™ formulation, are already widely used in the environmental and clinical setting, including wound care dressings and ophthalmic solutions. In a clinical review, PHMB was stated to have good clinical safety with no known toxic risks (Gray et al. 2010), and even with long-term use, BAC was stated to pose no clinical risk (Marple et al. 2004). Therefore, although cytotoxicity testing would need to be performed prior to a Byotrol™-coated catheter entering the clinical setting, it is probable that Byotrol™ should be safe to use.
In conclusion, Byotrol™ G32 has the ability to reduce bacterial growth as an antimicrobial in suspension, as a coating, and is also able to disrupt an existing biofilm. It may therefore hold great promise not only as an antimicrobial coating for medical devices but also as a sanitizing agent for the removal of established biofilms.
We would like to thank Age UK for funding this research and Byotrol Technology Ltd. for sample provision. The microscopes used in this study were part of The Bioimaging Facility at the University of Manchester, purchased with grants from BBSRC, the Wellcome Trust and the University of Manchester Strategic Fund.