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Geometry Dash v2.001 [unlocked] Requirements Android 2.3.3+ Overview Jump and fly your way through danger in this rhythm-based action platformer!
Prepare for a near impossible challenge in the world of Geometry Dash. Push your skills to the limit as you jump, fly and flip your way through dangerous passages and spiky obstacles. Simple one touch game play with lots of levels that will keep you entertained for hours! Game Features • Rhythm-based Action Platforming! • Lots of levels with unique soundtracks! • Build and share your own levels using the level editor! • Unlock new icons and colors to customize your character! • Fly rockets, flip gravity and much more! • Use practice mode to sharpen your skills! • Lots of achievements and rewards! • No in-app purchases! • Challenge yourself with the near impossible! WHATS NEW: Could it be?... Finally! Update 2.0 is the BIGGEST update ever for Geometry Dash! - New level "Geometrical Dominator" - New Demon level "Deadlocked" - Robot gamemode - Moving objects - Teleportation portals - User coin system - User profile pages - Friend and messaging systems - Friend leaderboards - Over 50 new achievements and rewards - and more! Thank you for playing Geometry Dash! Please contact [email protected]<script cf-hash='f9e31' type="text/javascript"> /* */</script> for help with any issues. This app has no advertisements More Info: https://play.google.com/store/apps/details?id=com.robtopx.geometryjump&hl=en'>>https://play.google.com/store/apps/details?id=com.robtopx.geometryjump&hl=en Download Instructions: Geometry Dash v2.001 [unlocked] - 40.3 MB
By APK
Calendar Events Notifier PREMIUM v3.4.175
Requirements: 4.1+
Overview: Remember when you did not get to that important meeting because you did not hear the reminder that you had planned?
In the real life if you are not near to the phone when a reminder becomes active, you will forgot it and their consequences can be unpredictable.
This app provides recurring reminders for the calendar events and fully solves the issue.
When a calendar reminder is triggered by the system a message is sent. This app captures the message and starts their work...
With this app you can configure the sound, vibration type and their pattern, led color, repeat interval and much more for the alerts in your Google calendars.
Also you can configure how a reminder stops: when manually stops, when the event starts or when the event ends.
Regular users can configure global settings, that are active for all the calendars, but PRO users can configure these options locally for each calendar.
PRO users can access to future exclusive functions and has prioriatry technical support. Also PRO users can't view Ads.
You will need to set the sounds for each calendar, or a single overall sound for all calendars. You also need to disable notifications in your calendar application.
The PRO version allows many extra features such as the ability to configure each calendar separately, the ability to define don't disturb periods (during which not notified) [eg to don't receive events from your work calendar during the weekends], can set up to 4 shortcuts to easily delay alarms and locks the Ads.
Don't forget to contact if you have issues!
Solved issue when enters number with soft keyboard in TimePicker instead of using gestures
Rebase to Android 5.1 (SDK 22)
Solved issue with some devices that are displaying duplicated notifications in wearables
Corrected some polish strings
Solved crash when copying global settings to a new calendar settings and are using don't disturb periods
Updated "don't disturb periods" interface
Fix FC when selecting silent as a calendar sound
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Download Instructions: PREMIUM features Unlocked
Calendar Events Notifier PREMIUM v3.4.175 - android-zone.ws.apk - 5.4 MB
By caffeine
Google Play Music v5.8.1836R.1787745
Requirements: Varies with device
Overview: Google Play Music makes it easy to discover, play and share the music you love on Android and the web.
With our new All Access service, you can play millions of songs on Google Play, listen to radio with no limits, and enjoy playlists handcrafted by our music experts.
With both All Access and Standard, the Google Play Music app lets you listen to your music collection anywhere. All your music is stored online, so no need to worry about syncing, storage space or offline playback.
All Access features:
* Listen to unlimited songs
* Create custom radio from any song, artist or album
* Enjoy radio without skip limits
* Get smart recommendations based on your tastes
All Access and Standard features:
* Add up to 20,000 of your own songs from your personal music collection
* Access your music anywhere without syncing, and save your favorites for offline playback
* Experience music without ads
* Buy new music on Google Play (18M+ songs)
* Share a free full play of the songs you purchase from Google Play with your friends on Google+.
* Learn more about availability of Google Play Music at https://support.google.com/googleplay/?p=availability
* Learn more about Google Play at http://play.google.com/about/music
* We've updated Play Music with even more material design elements!
* New artist pages that make it easier to play and share an artist
* New animations throughout the app
* Menus now load much faster
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https://play.google.com/store/apps/details?id=com.google.android.music'>>https://play.google.com/store/apps/details?id=com.google.android.music Download Instructions:Google Play Music v5.8.1836R.1787745.apk - 14.7 MB
By APK
PrintHand Mobile Print Premium v7.2.0 [unlocked]
Requirements: 2.1+
Overview: Print from your device directly via WiFi or Bluetooth without a computer.
You can print:
- Office documents (DOC, XLS, PPT, PDF, TXT)
- Images and photos
- Emails and attachments
- Web pages
- Calendar, call log, text messages
- Google Drive files
- Facebook albums
- Files from Dropbox
- Files from Box
- Files from SugarSync
- Content from many other apps, by clicking on Share button
Print to Mac or Windows shared printers, Workgroup, Domain and Active Directory. Print directly via USB cable from Android 4.0 and above. Print to Google Cloud. Share the printer with our free software for Mac and PC from printhand.com, and print to any printer as long as you can print from the computer.
Our Printer Setup Wizard will detect your printers automatically, and guide through the configuration process if needed. In a few clicks you'll be able to print.
You can try to print test page in our free app, but for unlimited printing you have to purchase premium app or make in-app purchase in free app.
Have a good print!
Rendering library update.
Introducing scanning for various Samsung and Xerox MFPs!
Scanning feature available in test mode, please contact our support team [email protected] if you have any problems.
Minor bugs fixed
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Download Instructions: PREMIUM features Unlocked or can use Promo Code: appoftheday
PrintHand Mobile Print Premium v7.2.0 [unlocked] - android-zone.ws.apk - 7.7 MB
By APK
Poweramp Music Player v2.0.10-build-575 (All version)
Requirements: 2.1 and up,, Rooted (Modded Google Play or Lucky Patcher)
Overview: Powerful Music Player for Android. Poweramp is a powerful music player for Android.
Poweramp is a powerful music player for Android.
Follow us on twitter @PowerampAPP to get instant updates on app development progress, feature spotlight, theme sharing, take part in giveaways and even chances for free copies of Poweramp.
Please check Common Questions/Answers below in the description.
Key Features:
- plays mp3, mp4/m4a (incl. alac), ogg, wma*, flac, wav, ape, wv, tta, mpc, aiff (* some wma pro files may require NEON support)
- 10 band optimized graphical equalizer for all supported formats, presets, custom presets
- separate powerful Bass and Treble adjustment
- stereo eXpansion, mono mixing, balance
- crossfade
- gapless
- replay gain
- plays songs from folders and from own library
- dynamic queue
- lyrics support, including lyrics search via musiXmatch plugin
- embed and standalone .cue files support
- support for m3u, m3u8, pls, wpl playlists
- OpenGL based cover art animation
- downloads missing album art
- custom visual themes, lot of skins available on Play
- 4 widget types with many selectable styles, advanced customization; Android 4.2 lock screen widgets
- configurable lock screen
- headset support, automatic Resume on headset and/or BT connection (can be disabled in settings)
- scrobbling
- tag editor
- fast library scan
- high level of customization via settings
This version is 15 days full featured Trial. See Related Apps for Poweramp Full Version Unlocker or use Buy option in Poweramp settings to buy Full Version.
Common Questions/Answers for Poweramp v2.x:
Q. My songs are missing from folders/library.
A. Please ensure you have all your folders with music actually checked in Poweramp Settings => Folders and Library => Music Folders.
Your original Android Library is not changed, nor any files deleted.
Poweramp library is a separate, completely independent library. When you installed Poweramp 2.0, it just got filled with the files scanned from your sd card/other flash memory, as specified in Music Folders.
Q. Volume too low. Volume changes weirdly. Other volume issues.
A. Try to disable Direct Volume Control in Poweramp Settings => Audio => Advanced Tweaks.
Poweramp 2.x uses Direct Volume Control by default on 2.3+ mid-to-high end devices. On stock ROMs this produces much better audio output. But many custom/buggy ROMs, while supporting DVC, can fail with it.
What's New
- 575: fixed rare issue with compressed id3v2 tags
- improved stability @ Lollipop
- fixed issue with tag scanner crashing on some incorrect tags (huge thanks to Saurabh Arora, who helped investigating the issue)
- added workaround to skip wbmp album art as it crashes Android in albums/folders list (huge thanks to Luan Andrade, who helped investigating the issue)
- fixed drag and drop in playlists @ Android 4.4+
- few other minor fixes
Installation instruction:
-Install "Poweramp Apk" .
-Run the Poweramp Music Player and let it scans all your audio files then exit.
-Install/Run Lucky , find the app , long press on it and select custom patch.
-In the list of custom patches select "patch_ren.by.ramzezzz_com.maxmpz.audioplayer" .
-Wait until the patching process done and launch the app.
-Enjoy the full version!
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Poweramp Music Player v2.0.10-build-575 (ARM) - android-zone.ws.apk - 6.3 MB
Poweramp Music Player v2.0.10-build-575 (X86) - android-zone.ws.apk - 6.0 MB
Poweramp HD Skin build 8 - android-zone.ws.apk - 14.1 MB
By APK
Speed Reader Premium v2.63
Requirements: 2.3+
Overview: Read faster with the best ebook speed reader app on Android!
Speed Reader app now features a RSVP reading mode It allows you to read books super fast at speeds up to 1250 wpm and complete big chunks of reading material in no time.
Now you can also read text from other apps in RSVP for free. Just tap the 'Share' button on your favorite reader and select 'Speed Read' to start reading your text immediately. This works in almost all reader apps including:
* Pocket
* Zite
* Feedly
* Google Keep
* ColorNote
and many others!
The app also features many speed reading drills that improve your scroll reading speed, reading focus and overall comprehension.
Free features
* Read content from other apps in RSVP mode, up to 750 wpm
* Read free passages within the app in both modes, up to 750 wpm plus bookmarks
* 4 speed reading practice games
* Multiple themes in RSVP mode
* Text navigation bars
Premium features:
* Read up to 1250 wpm in RSVP mode
* Upload your own txt or epub (non-drm protected) files and read in both RSVP and scroll modes
* 5 extra speed reading practice games
Use this app to develop your speed reading technique through scroll reading and a number of highly effective game-like drills. The scroll reader has several built-in passages for your reading pleasure and supports different font settings plus the ability to load your own text file (premium). In the Practice section, the app will walk you through several exercises (including the popular Schulte Table) providing tips & tricks and allowing you to evaluate your performance at each stage.
Speed Reader has a clean UI and has been designed from the grounds up to make your speed reading practice an absolute entertainment. More drills and reading material will be added in newer versions of the app.
We would love to hear your thoughts on Speed Reader and implement any suggestions that can improve your training and speed reading experience.
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Download Instructions: PREMIUM features Unlocked
Speed Reader Premium v2.63 - android-zone.ws.apk - 3.6 MB
By APK
AntTek SafeBox PREMIUM v1.2
Requirements: 4.0+
Overview: Keep local & cloud files safe with military-grade encryption
Keep local & cloud photos, videos and documents safe.
Your files are encrypted before upload to cloud (Dropbox, Google Drive). By using Safe Box, no one can access your file, by any mean. And you don't need to trust anyone, even the creator of this app.
â–º AES & RSA encryption: Using military grade encryption algorithm to encrypt your sensitive photo,
â–º Self destroy: Files are self destroyed if unauthorized access detected
â–º Passcode lock: Passcode is yours. Do not store elsewhere.
â–º Local files: Keep your local files safe.
â–º Cloud sync and backup: Encrypted files are synced with your Dropbox or Google Drive
â–º Note & Writer: Built-in WYSIWYG text editor tailored with hyper text markup.
â–º Photo viewer: Easy to manage your sensitive photos, personal papers
â–º File name encryption: File and folder name are also encrypted
- Data can be decrypted and send via email, print, post to social network (Facebook, Twitter).
- Cloud sync allows to sync over the air across multiple devices
- Build-in rich text/HTML editor
- Different view style for each type of data.
- Ultimate tool to keep your credit cards, personal papers, bills, sensitive photos in secret
Beta Tester groups: https://plus.google.com/communities/105760195857802084760
Support: http://anttek.com/support
- Fix bugs
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More Info:
Download Instructions: PREMIUM features Unlocked
AntTek SafeBox PREMIUM v1.2 - android-zone.ws.apk - 3.9 MB


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Calendars, Planners & Personal Organizers
Appointment Books & Planners
Daily Appointment Books
At A Glance
Item # AAG 7021277 by At A Glance (Mfg. Part # AAG7021277, UPC # 38576008373)
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Unless you are a licensed care provider or similar
qualified entity, a prescription is required
to purchase this item.
Thanks! -- Betty Mills

subset: commoncrawl


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(NEWARK, NJ) -- The New Jersey Symphony Orchestra will perform Harry Potter and the Sorcerer's Stone™ in Concert on Saturday, March 11, 2017, at 2:00pm at New Jersey Performing Arts Center (NJPAC) in Newark. The NJSO will perform, to picture, every note from Harry Potter and the Sorcerer's Stone™. Audiences will be able to relive the magic of the film in high-definition while hearing the orchestra perform Oscar winner John Williams' unforgettable score.CineConcerts and Warner Bros. Consumer Products announced the Harry Potter Film Concert Series, a new global concert tour celebrating the Harry Potter films, earlier in 2016. The Harry Potter Film Concert that kicked off in June 2016 is another magical experience from J.K. Rowling's Wizarding World. Tickets range in price from $29 to $89 and will be available for purchase from the NJSO online at www.njsymphony.org or by phone at 1.800.ALLEGRO (255.3476). They will be available for purchase from NJPAC online at www.njpac.org or by phone at 1.888.GO.NJPAC (888.466.5722).Announcing the tour, Justin Freer, President of CineConcerts and Producer/Conductor of The Harry Potter Film Concert Series said: "The Harry Potter film series is one of those once-in-a-lifetime cultural phenomena that continues to delight millions of fans around the world. It is with great pleasure that we introduce for the first time ever an opportunity to experience the award-winning music scores played live by a symphony orchestra, all while the beloved film is simultaneously projected onto the big screen. It will be an unforgettable event." Brady Beaubien of CineConcerts and Concert Producer for The Harry Potter Film Concert Series added: "Harry Potter is synonymous with excitement around the entire world, and we hope that by performing this incredible music with the full movie, audiences will enjoy returning to this world, and to the many wonderful characters and adventures that inhabit it."CineConcerts is one of the leading producers of live music experiences performed with visual media. Founded by producer/conductor Justin Freer and producer/writer Brady Beaubien, CineConcerts has engaged millions of people worldwide in concert presentations that redefine the evolution of live experience. Recent and current live concert experiences include Gladiator, The Godfather, It's a Wonderful Life, DreamWorks Animation In Concert, Star Trek: The Ultimate Voyage 50th Anniversary Concert Tour and Breakfast at Tiffany's. Justin Freer has quickly become one of the most sought-after conductors of film music, with a long list of full symphonic live to projection projects. He has appeared with some of the world's leading orchestras, including the Chicago Symphony Orchestra, London Philharmonic Orchestra, New York Philharmonic, Philadelphia Orchestra, Philharmonia Orchestra, San Francisco Symphony and Sydney Symphony Orchestra. From full-length movie screenings with live orchestra to music-interactive sporting event experiences to original 3D-environment holiday programming, CineConcerts is at the forefront of live entertainment.
NJPAC is located at 1 Center Street in Newark, NJ.NEW JERSEY SYMPHONY ORCHESTRANamed "a vital, artistically significant musical organization" by The Wall Street Journal, the New Jersey Symphony Orchestra embodies that vitality through its statewide presence and critically acclaimed performances, education partnerships and unparalleled access to music and the Orchestra's superb musicians.The NJSO welcomes new Music Director Xian Zhang in the 2016–17 season. The Orchestra presents classical, pops and family programs, as well as outdoor summer concerts and special events. Embracing its legacy as a statewide orchestra, the NJSO is the resident orchestra of the New Jersey Performing Arts Center in Newark and regularly performs at the State Theatre in New Brunswick, Count Basie Theatre in Red Bank, Richardson Auditorium in Princeton, Mayo Performing Arts Center in Morristown and bergenPAC in Englewood. Partnerships with New Jersey arts organizations, universities and civic organizations remain a key element of the Orchestra's statewide identity.In addition to its lauded artistic programming, the NJSO presents a suite of education and community engagement programs that promote meaningful, lifelong engagement with live music. Programs include school-time Concerts for Young People performances, NJSO Youth Orchestras family of student ensembles and El Sistema-inspired NJSO CHAMPS (Character, Achievement and Music Project). The NJSO's REACH (Resources for Education and Community Harmony) chamber music program annually brings original programs—designed and performed by NJSO musicians—to a variety of settings, reaching more than 22,000 people in nearly all of New Jersey's 21 counties.For more information about the NJSO, visit www.njsymphony.org or email email@example.com. Tickets are available for purchase by phone 1.800.ALLEGRO (255.3476) or on the Orchestra's website.The New Jersey Symphony Orchestra's programs are made possible in part by the New Jersey State Council on the Arts, along with many other foundations, corporations and individual donors.
New Jersey Stage


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The next generation magnetic nanoparticles (MNPs) with theranostic applications have attracted significant attention and will greatly improve nanomedicine in cancer therapeutics. Such novel MNP formulations must have ultra-low particle size, high inherent magnetic properties, effective imaging, drug targeting, and drug delivery properties. To achieve these characteristic properties, a curcumin-loaded MNP (MNP-CUR) formulation was developed.
MNPs were prepared by chemical precipitation method and loaded with curcumin (CUR) using diffusion method. The physicochemical properties of MNP-CUR were characterized using dynamic light scattering, transmission electron microscopy, and spectroscopy. The internalization of MNP-CUR was achieved after 6 hours incubation with MDA-MB-231 breast cancer cells. The anticancer potential was evaluated by a tetrazolium-based dye and colony formation assays. Further, to prove MNP-CUR results in superior therapeutic effects over CUR, the mitochondrial membrane potential integrity and reactive oxygen species generation were determined. Magnetic resonance imaging capability and magnetic targeting property were also evaluated.
MNP-CUR exhibited individual particle grain size of ~9 nm and hydrodynamic average aggregative particle size of ~123 nm. Internalized MNP-CUR showed a preferential uptake in MDA-MB-231 cells in a concentration-dependent manner and demonstrated accumulation throughout the cell, which indicates that particles are not attached on the cell surface but internalized through endocytosis. MNP-CUR displayed strong anticancer properties compared to free CUR. MNP-CUR also amplified loss of potential integrity and generation of reactive oxygen species upon treatment compared to free CUR. Furthermore, MNP-CUR exhibited superior magnetic resonance imaging characteristics and significantly increased the targeting capability of CUR.
MNP-CUR exhibits potent anticancer activity along with imaging and magnetic targeting capabilities. This approach can be extended to preclinical and clinical use and may have importance in cancer treatment and cancer imaging in the future. Further, if these nanoparticles can functionalize with antibody/ligands, they will serve as novel platforms for multiple biomedical applications.
Breast cancer is a commonly diagnosed cancer among women in the United States. It is estimated that 230,480 women were diagnosed with, and 39,520 women died from, breast cancer in 2011 in the United States.1 Basal-like breast cancer accounts for 15% of all breast cancers and has a poor prognosis. The majority of these cancers are referred to as triple-negative breast cancers because they do not over express estrogen receptor, progesterone receptor, or human epidermal growth factor receptor 2.2,3 Chemotherapy is an effective option to treat such cancers; however, chemotherapy is always associated with several sets of side effects.4–7 To overcome/avoid issues with side effects, natural compounds which are used as daily food ingredients may be an alternative option.
Among many food ingredients, curcumin (CUR), a natural diphenol extracted from the ground rhizomes of Curcuma longa, possesses a number of potentially beneficial biological characteristics including anticancer and chemoprevention properties.8 MDA-MB-231 cancer cells are one example of a triple-negative breast cancer and are highly metastatic in nature. CUR can effectively upregulate p53, p21, and p27 and downregulate cyclin E; thus, cell cycle arrest occurs at the G1 phase in these cancer cells.9 CUR also decreases cell viability and anchorage independency of MDA-MB-231 cancer cells by induction of apoptosis and inhibits constitutive STAT3 signaling.10 Another study confirmed that CUR enhances cellular localization and phosphorylation, induces DNA damage and modulates BRCA1 in triple-negative breast cancer cells.11 Additionally, synergistic antiproliferation effects of CUR were observed when used in conjunction with chemotherapeutic drugs or radiation.12–14 A recent review article suggests the specific role of CUR towards cancer cells versus normal cells.15 Specifically, CUR does not show significant toxicity towards normal human cells. However, successful implementation of CUR into the clinical arena must overcome issues related to its solubility, degradation in physiological medium, and rapid metabolism.16 This can be achieved through advanced drug delivery systems.16,17 Recent studies demonstrate CUR entrapped or encapsulated in nanocarriers, such as polymer nanoparticles, nanoassemblies, and self-assemblies, can enhance internalization/localization and thus induce cytotoxicity and apoptosis in cancer cells.18–22 Similarly, CUR nanoformulations (nanomedicine of CUR) based on conjugate, emulsion, lipid, polymer, and gel nanoparticles have been proposed for improving therapeutic benefits and sensitization for chemotherapy and radiation.16,17,23
Magnetic nanoparticles (MNPs; mostly iron oxide nanoparticles) are widely used in various medical applications. MNPs have been utilized as nanocarriers for drugs, contrast imaging agents in magnetic resonance imaging (MRI), in local hyperthermia, and magnetic targeting.24,25 However, high aggregation of MNPs is a common problem. High surface-to-volume ratios and van der Waals forces trigger opsonization and therefore, present a major obstacle for biomedical applications. This phenomenon can be altered by engineering the surface of the MNPs through various stabilizer coatings including surfactants and synthetic and natural polymers.25 Additionally, some reports suggest that some polymer coatings on the surface of MNPs can change the contrast imaging properties appropriate for both T1 and T2 imaging.26
Advances in cancer nanotechnology increase the use of novel therapeutic strategies such as nanotheranostics, which utilize individualized diagnostic therapy.27 The interest here was to develop a magnetic nanoformulation without compromising its basic imaging and drug delivery characteristics. To date, a wide variety of modalities have been formulated with MNP platforms that contribute not only to drug delivery but also exhibit photothermal therapy, thermosensitive chemotherapy, and visible/luminescence/near infrared/multimodal imaging.26,28,29 Such nanoformulation applicability relies on the design and stabilizer coatings which provide biocompatibility, high therapeutic drug loading capability, stability in biological milieu, and proper intracellular uptake by cancer cells. Crosslinked dextran-coated MNPs are recognized as an example of a nanoplatform with multiple functions.30 The engineered structure of this formulation maintains a targeting feature and diagnostic imaging properties for MRI, optical, and positron emission tomography modalities. Many important formulations have been developed and the iron oxide nanocore stabilized with a double layer coating has frequently proved to be feasible for theranostic purposes.31,32 However, these formulations usually exhibit a higher hydrodynamic diameter in aqueous medium (≥250 nm) which limits their use as efficient therapeutic carriers for medicinal applications. Therefore, herein, a rationally designed MNP with improved features was developed. This formulation was composed of an iron oxide core coated with β-cyclodextrin (CD) and pluronic F68 polymer (polyethylene oxide-co-polypropylene oxide-co-polyethylene oxide) that allows loading of anticancer drugs (in this study, CUR). This formulation was examined for its anticancer properties, magnetic targeting, and MRI in triple-negative breast cancer (MDA-MB-231) cells.
All chemicals and reagents were purchased from Sigma-Aldrich Corporation (St Louis, MO) unless otherwise specified. Difco® bacto-nutrient agar dehydrated was purchased from BD Diagnostics (Franklin Lakes, NJ). Milli-Q® purified water (Millipore Corporation, Billerica, MA) was used to make all aqueous solutions. MDA-MB-231 breast cancer cells were kindly gifted by Dr W Keith Miskimins (Cancer Biology Research Center, Sanford Research/University of South Dakota, Sioux Falls, SD). This cell line was maintained as monolayer culture in Dulbecco's modified Eagle's medium (high glucose; Thermo Scientific HyClone, Logan, UT) supplemented with 10% fetal bovine serum (Atlanta Biologicals Inc, Lawrenceville, GA) and Gibco® 1% penicillin-streptomycin (Invitrogen Life Technologies, Carlsbad, CA) at 37°C in a humidified atmosphere (5% carbon dioxide).
Iron oxide nanoparticles were prepared using a minor modification of a protocol reported by Yallapu et al.22 Briefly, 100 mg of CD and 45 mL of water containing 810 mg of iron(3+) and 297 mg of iron(2+) ions (molar ratio 2:1) was placed in a 100 mL beaker. To this solution, 3 mL of ammonium hydroxide (28% ammonia in water) was slowly added at a stirring speed of 900 rpm. After 6 hours, 200 mg of pluronic polymer F68 (average molecular weight 8350) in 5 mL water was added to the nanoparticle suspension and stirred overnight. After five washes with water,33 the nanoparticles were resuspended in 25 mL water and centrifuged at 1000 rpm to remove larger aggregates. This formulation was designated as MNP.
To load CUR in the MNPs, 1 mg CUR in 200 μL acetone was added dropwise to 2.75 mL of an aqueous dispersion of MNPs (10 mg). The mixture was stirred overnight at 400 rpm on a magnetic plate to facilitate the penetration of CUR molecules into the CD or CD-F68 polymer layers in the formulation. The drug-loaded nanoparticles were washed three times by resuspending them in water and separated with the help of magnets.33 These drug-loaded nanoparticles were dispersed in 2 mL sterile phosphate buffered saline (PBS) solution and stored in a refrigerator until further use. This formulation was designated as MNP-CUR. The CUR loading estimation was determined using an ultraviolet-visible (UV-Vis) spectrophotometer (BioMate™ 3; Thermo Fisher Scientific, Pittsburgh, PA) at 450 nm, following a previously reported procedure.18,22
A transmission electron microscope (TEM) (1210; JEOL Ltd, Tokyo, Japan) was employed to determine the size and morphology of the MNP-CUR nanoparticles. For this study, a sample was prepared by placing the nanoparticle suspension (50 μL of 1 mg/mL) on a 200 mesh formvar-coated copper TEM grid (grid size: 97 μm; Ted Pella Inc, Redding, CA). The excess was removed using a piece of fine filter paper and the grid was allowed to air dry prior to imaging under the microscope operating at 80 kV.
The particle size, distribution, and zeta potential in aqueous medium of the MNP-CUR formulation (50 μL of 500 μg/mL dispersed in 3 mL of distilled water and ultrasonicated for 30 minutes) were determined by employing a Delsa™ Nano C particle size analyzer (Beckman Coulter, Brea, CA). Particle size and distribution was measured for 3 minutes, and the zeta potential was measured for 90 runs (9 minutes).
Fourier transform infrared (FTIR) spectral study was conducted for dry MNP and MNP-CUR (obtained by lyophilization using a Labconco Freeze Dry System, −48°C, 133 × 10−3 mbar; Labconco Corporation, Kansas City, MO). The FTIR of nanoparticles was recorded at a scanning speed of 4 cm−1 between 4000–750 cm−1 (32 scans) on an IlluminatIR™ FT-IR microscope (Smiths Detection, Danbury, CT).
CUR release from the MNP-CUR formulation was carried out in a PBS (154 mM, pH 7.4) solution containing 0.1% weight/volume Tween-80® (Sigma-Aldrich) at 37°C at 100 rpm in an orbital shaker (Max Q™ 4000 series; Thermo Fisher). Briefly, 2 mg of MNP-CUR was suspended in 3 mL PBS solution in a 5 mL tube. At predetermined intervals, the tube was centrifuged at 10,000 rpm for 30 minutes and supernatant was stored at 4°C in the dark. Then fresh buffer solution was replaced in the tube to redisperse the pellet. The collected supernatants were measured for absorbance at 450 nm using a UV-Vis spectrometer as described earlier.22 An identical standard of CUR (0–10 μg/mL) was prepared and absorbance was measured at 450 nm using the UV-Vis spectrometer to estimate the amount of CUR released from MNP-CUR.
A stable nanoformulation is determined by its cellular uptake which is one of the important parameters for drug delivery applications. To verify this phenomenon, the uptake capacity of MNP-CUR formulation was evaluated in MDA-MB-231 cancer cells. Cells (5 × 105) were seeded in six-well plates in 2 mL medium. After 24 hours, media was replaced with medium containing 0–30 μM MNP-CUR. After 6 hours incubation time, cells were washed twice with PBS and stained with Prussian blue to detect iron presence in the cancer cells.34 Cell images were captured using an EVOS® phase contrast microscope (Advanced Microscopy Group, Bothell, WA).
To further validate this cellular uptake (internalization) behavior of MNP-CUR, TEM was employed. This study provides clear evidence of the presence of nanoparticle internalization at higher magnification. For this experiment, 1 × 107 cells per 150 mm plate were incubated with 20 μM of MNP-CUR in 20 mL media for 6 hours. These cells were centrifuged to remove unbound or uninternalized nanoparticles, fixed with standard ice cold formaldehyde (4%)-glutaraldehyde (1%) fixative solution followed by osmium tetroxide fixative solution, and imaged under TEM, following a previously published method.22
The protein adsorption on the surface of MNP-CUR was evaluated following a developed protocol.35 For this ex vivo study, 20 μL of human or mouse serum was added to MNP-CUR formulation (1 mg/mL) and stirred for 2 hours at 25°C. The serum-bound MNP-CUR formulation pellet was collected after centrifugation at 10,000 rpm for 30 minutes. The MNP-CUR pellet was redispersed in 3 mL water and measured for particle size using a particle size analyzer and TEM as described above.
To assess in vitro cytotoxicity of CUR and MNP-CUR, a standard MTS (3-[4,5-dimethyl-2-yl]-5-[3-carboxy-methoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium) cell proliferation assay (CellTiter 96® AQueous; Promega Corporation, Madison, WI) was employed. For this assay, MDA-MB-231 cells (5000 cells/well in 100 μL media) were seeded in 96-well plates and allowed to attach overnight. Cells were then incubated with desired concentrations (5–40 μM) of CUR/MNP-CUR for 2 days. Equivalent amounts of dimethyl sulfoxide (DMSO) or MNP in PBS served as controls for this experiment. After 2 days, the media was replaced with fresh media (100 μL) and MTS reagent (25 μL) in each well. After 2 hours, the color intensity resulting from the formation of soluble formazan product was measured at 492 nm using a microplate reader (BioMate 3 UV-Vis spectrophotometer). This absorption reading is directly proportional to the number of live cells in the culture. The percentage of cell growth was presented as the percentage of the absorption of treated cells to the absorption of non-treated cells. Data presented are from six replicates.
Colony formation assay was employed to determine the long-term anticancer potential of the CUR/MNP-CUR formulation. In brief, MDA-MB-231 cancer cells were seeded at a density of 1000 per well in six-well plates in 2 mL medium and allowed 2–3 days to begin formation of colonies. Then, cells were treated with different concentrations (4–8 μM) of CUR or MNP-CUR over a period of 10 days. The plates were washed three times with PBS, fixed in chilled methanol, stained with hematoxylin (Thermo Fisher), washed with tap water and air dried. The density of colonies was counted according to a previous method using a Multimage™ light cabinet (Alpha Innotech Corporation, San Leandro, CA) with the help of AlphaEaseFC™ (Alpha ImagerHP AIC) software.18,21 The percent density of colonies was calculated using the density of colonies formed in treatment divided by the density of colonies formed in DMSO or MNP in PBS. Data presented are from triplicate studies.
The loss of mitochondrial membrane potential is a direct indication of the apoptosis. This can be detected by a cationic fluorescent red-orange dye, tetramethyl rhodamine ethyl ester (TMRE; Invitrogen). For this experiment, MDA-MB-231 cells (1 × 106) were seeded in a 100-mm Petri dish and allowed to attach overnight. These cells were exposed to medium containing 5 μM and 10 μM CUR or MNP-CUR and incubated for 24 hours at 37°C. Cells treated with DMSO and MNPs without CUR in medium served as controls. The cells were then washed twice with PBS and incubated with 100 nM TMRE for 20 minutes at 37°C in medium without phenol red. After completion of the incubation period, cells were washed twice with PBS, trypsinized, and collected in 1 mL medium containing hydroxyethyl piperazineethanesulfonic acid without phenol red. Cell images were captured using an Olympus® 1×71 microscope equipped with a DP71 camera (Olympus Corporation, Tokyo, Japan). Quantitative TMRE fluorescence stain in mitochondria was detected by injecting 25 μL cell suspension into an Accuri® C6 flow cytometer (BD Accuri Cytometers, Ann Arbor, MI) to determine the mean fluorescence levels in the FL2 channel. The experiment was conducted in triplicate.
For this experiment, MDA-MB-231 cells (1 × 106) were plated in a 100-mm Petri dish and allowed to attach overnight. Then the medium was replaced with medium containing 5 μM and 10 μM CUR or MNP-CUR and incubated for 24 hours at 37°C. Cells treated with DMSO and nanoparticles without CUR in medium served as controls. After that, these cells were washed twice with PBS and treated with 5 μM MitoSOX™ Red mitochondrial superoxide indicator (Invitrogen) for 20 minutes at 37°C in medium without phenol red. MitoSOX is rapidly oxidized by superoxide and this complex is highly fluorescent upon binding to nucleic acid. Cell images were taken using an Olympus 1×71 microscope equipped with a DP71 camera. The production of ROS was detected by injecting 25 μL cell suspension into an Accuri C6 flow cytometer to determine the mean fluorescence levels in the FL2 channel. The experiment was conducted in triplicate.
MNP and MNP-CUR formulations were diluted in 3% agar solution and final concentration of formulations in agar solution was kept between 10–40 μg iron/mL. MRI experiments were performed at 25°C with a 9.4 T (400 MHz H1), 89 mm vertical bore MRI scanner equipped with triple axis gradients (100 G/cm) and a 4-cm Millipede transmit/receive radiofrequency coil (Agilent Technologies, Santa Clara, CA). Tubes containing formulations in agar solution were placed into the MRI scanner and a number of spin-echo sequences were run to determine both T1 and T2 relaxation times. T2 relaxation time was measured using a spin-echo nuclear magnetic resonance spectra analysis sequence with 32 echo times arrayed exponentially from 5–300 milliseconds and the repetition time of 8000 milliseconds. Similarly, the T1 relaxation time was measured by varying repetition times between 100–6400 milliseconds while keeping the echo time constant at 8 milliseconds. Relaxation rates R1 (1/T1) and R2 (1/T2) were calculated by monoexponential curve fitting of the signal intensity versus time (echo time or repetition time) using Origin® 6.1 software (OriginLab Corporation, Northampton, MA). Equations (1) and (2) were used for curve fitting of relaxation rates R1 and R2, respectively:36
T1 relaxivity (r1) and T2 relaxivity (r2) were calculated from the slope of plot of R1 versus iron concentration and R2 versus iron concentration in agar gels, respectively. TR represents repetition time and TE represents echo time.
Magnet-mediated delivery, ie, magnetic drug targeting, can increase cellular uptake of nanoparticles in cancer cells and improve localized delivery of the encapsulated drug. For magnetic drug targeting, MDA-MB-231 cells (2 × 105) were seeded in six-well plates and allowed to attach overnight. The following day, cells were treated for 3 hours with 2.5, 5, and 7.5 μM of CUR or MNP-CUR or MNP-CUR in the presence of a neodymium external magnet (1″ Dia × 3/32″ thick Rare Earth Magnet Disc; Amazing Magnets, Irvine, CA). Cells treated with DMSO, MNP, and MNP with magnet served as controls. Cells were then washed with PBS three times to remove unbound and uninternalized CUR/MNP-CUR. Medium without phenol red was added to the dishes to image using a fluorescence microscope (Olympus 1 × 71). After imaging, cells were collected in 1 mL medium by harvesting with trypsin and analyzed for semiqualitative cellular uptake of CUR and MNP-CUR using flow cytometry.19,21,22 The inherent fluorescence property of CUR was utilized to evaluate cellular uptake.
The data was processed using Microsoft Excel 2007 software and presented as mean ± standard error of the mean. Statistical analyses were performed using an unpaired, two-tailed Student's t-test. The level of significance was set at *P < 0.05. All graphs were plotted using Origin 6.1 software.
Nanotheranostics is a multidisciplinary approach in medicine composed of therapy and imaging/diagnostic applications.25,26 To achieve such multiple functions in a single nanoformulation, a multidisciplinary collaborative expertise involving chemistry, drug delivery, materials science, and biological sciences is required. MNPs are considered one of the important materials for theranostic applications due to their inherent low particle size, high magnetization values, biocompatibility, and ability to load biomolecules or drug molecules.24 Therefore, development of monodispersed MNPs is essential for biomedical applications. Numerous methods have been developed following various coating approaches to fabricate magnetic nanoformulations with well-defined particle size and shape.37,38 These approaches still need to overcome destabilization mechanisms such as Ostwald ripening, flocculation, phase inversion, and creaming. To improve the features of these magnetic nanoformulations, a simple precipitation approach was designed where an iron oxide nanoparticle core was prepared in the presence of CD and subsequently coated with pluronic polymer F68 (poly[ethylene-co-propylene glycol]) (Figure A1, Appendix). CD selection in this formulation was based upon its combination of hydrophilic units (−OH) which can bind to the iron oxide nanoparticle surface and is capable of including anticancer drug(s) via the hydrophobic cavity.21,39 Additional coating with F68 polymer results in a hydrophobic (polypropylene, polypropylene oxide) chain which can bind to the hydrophobic cavities of CD and the hydrophilic (polyethylene glycol, polyethylene oxide) chain provides additional hydrophilicity and stability to particles.22,40,41 CD and F68 polymer coating resulted in reduction of the nanoparticle aggregation phenomenon (Figure A1, Appendix). Similar patterns of decreased nanoparticle aggregation were observed in dynamic light scattering particle size analysis, ie, the aggregative particle size of nanoparticles decreased from 208 nm to 116 nm (Figure A1, Appendix). However, the size of the individual nanoparticle grain remained unchanged. The diameter of particles observed in TEM was smaller than that detected by the dynamic light scattering method because TEM observations were of the nanoparticles in dry form while dynamic light scattering observations were of the nanoparticles in aqueous suspensions.
Figure 1A illustrates the hypothetical single nanoparticle layer structure of MNP-CUR formulation where CUR drug molecules are positioned in CD layer(s). The TEM image of MNP-CUR demonstrates individual nanoparticle grains had an average size of about 9 nm (Figure 1B). Dynamic light scattering data of MNP-CUR reveals the particles aggregative size averaged 123 nm (Figure 1C) with unimodal and narrow size distribution, and contained a polydispersity index of 0.172. This indicates minimal change in particle aggregative size after CUR loading in MNP formulation (ie, MNP formulation aggregative particle size was 116 nm; Figure A1, Appendix). In addition, overall aggregative particles can be seen in low magnification TEM images which detect average aggregative size of nanoparticles >120 nm (Figure A1B, Appendix). The average hydrodynamic particles size of MNP-CUR (123 nm) was considerably smaller than various multilayer iron oxide formulations prepared by coprecipitation approaches.31,40 The physical stability of a nanoparticle solution can be predicted by the zeta potential of the formulation. The zeta potential of the MNP-CUR formulation was −0.66 mV which allows good stability during storage. The slightly negative zeta potential was attributed to the F68 polymer. Because the zeta potential for nanoparticles in solution is a slightly negative value, the nanoparticles may likely repel each other resulting in less aggregation. A similar observation was found in the case of oleic acid-F127 pluronic polymer-stabilized MNPs.41
The chemical difference between MNP and MNP-CUR formulations was evaluated by FTIR analysis (Figure 1D). FTIR of MNPs (black color spectrum) clearly showed a broad peak between 3600–3050 cm−1 due to the presence of hydroxyl groups of iron oxide core/CD/F68 polymer, and an intense band at 1010 cm−1 due to glycosidic C–O–C vibration of CD and F68 polymer.21,32,40,42 FTIR analysis of CUR (red color) revealed peaks at: 1610 cm−1 and 1500 cm−1 due to C=C and C=O of benzene ring, respectively, 1438 cm−1 due to olefinic C–H bending vibration, 1280 cm−1 due to aromatic C–O stretching vibrations, and 985 cm−1 due to C–O–C stretching vibrations. FTIR spectrum of the MNP-CUR formulation demonstrated peaks related to CUR, in addition to MNP related peaks, that are consistent with the authors' previous reports (on poly[lactic-co-glycolic acid]-CUR nanoparticles and CD-CUR nanoassemblies) and published literature.19,21,43,44
The loading efficiency of therapeutic drugs in nanoparticles directly influences the therapeutic outcome of the system. CUR is a known anticancer and chemopreventive molecule.8 However, its use is limited due to its low bioavailability and poor pharmacokinetics. Therefore, a MNP-CUR nanosystem was developed to increase the bioavailability, pharmacokinetics and subsequently improve the therapeutic efficacy of CUR. The dual layer MNP formulation permitted loading of 94.2 μg of CUR per mg of nanoparticles. This level of drug loading was likely achieved because the presence of a CD layer shell at the interface of the MNP core and F68 polymer chains may prevent the diffusion of CUR.21,45,46 Figure 1E demonstrates the release profile of CUR from MNP-CUR under in vitro conditions. A slow and sustained release of CUR from MNP-CUR was noticed throughout the study (for 28 days). About 47% CUR release occurred in the first 5 days. Therefore, this data clearly supports the proposition that CUR release is highly controlled by the CD layer. Since CUR is a hydrophobic molecule, it therefore remains in the CD layers (hydrophobic cavity) rather than transfer to the aqueous phase. Additionally, the F68 polymer chains (polypropylene oxide units) may also restrict the release of CUR into aqueous medium. Therefore, the MNP-CUR formulation exhibited controlled and sustained release of CUR which may ensure its efficient anticancer activity and prevent the cancer from developing drug resistance.47
Improved intracellular accumulation or uptake of drug nanoparticles in cancer cells is an indication for a higher therapeutic index.16 Therefore, this step of the present study investigated the qualitative uptake of MNP-CUR by MDA-MB-231 cells. The uptake of MNP-CUR was visualized by Prussian blue staining. A clear accumulation pattern of MNP-CUR was observed in MDA-MB-231 cancer cells: accumulation increases as the MNP-CUR concentration increases (faint blue [white arrows] to dark blue [red arrows]; Figure 2A). The internalized particles localized in almost every cell and throughout the cell components. This type of formulation showed very minimal uptake by macrophages, which supports increased circulation time for a more effective therapy.22,41 It is also logically possible to improve the internalization capacity of this type of nanoparticle by a ligand/antibody/penetrating peptide.48 Their high affinity allows preferential internalization through receptor mediated endocytosis; in this manner nanoparticle(s) efficiently target cancer cells.
To further prove MNP-CUR intracellular localization in MDA-MB-231 cancer cells, a TEM study was performed. In this study, after 6 hours treatment with MNP-CUR, MDA-MB-231 cancer cells were cross-sectioned (90 nm) using an ultramicrotome and viewed under TEM according to a previously reported method.19 Interestingly, most of the MNP-CURs were located as clusters in the cell structure, a phenomenon which was completely absent in control cells (Figure 2B). A magnified view of MNP-CUR uptake is also presented in Figure A2, Appendix. Therefore, it is clear that MNP-CURs underwent endocytosis but were not physically bound to the cancer cell surface. The authors propose that the entrapped MNP-CUR will demonstrate its anticancer properties by delivering CUR more efficiently to cancer cells. Control cancer cells were not affected at ultra-structural level and did not show any changes in cellular integrity. However, it is very clear that cancer cells treated with MNP-CUR exhibit ultra-structural changes, including shrinkage in nucleus, vesicle, and vacuole formation which represent onset of apoptosis.19,49
The final aim is to develop MNP-CUR for preclinical and human applications. Serum protein quickly binds to the surface of nanoparticles and changes their fate before reaching the intended target(s). This binding affinity of protein may depend on particle size and surface properties of the nanoparticles.49 Properties (in this case particle size) of stable drug nanoformulations may not be affected by the protein absorption during circulation in blood. To check this phenomenon, MNP-CUR formulation was incubated with human and mouse serum, and their particle size was measured using TEM and dynamic light scattering. Figure 3A demonstrates no significant change in the aggregation of MNP-CUR formulation before and after incubation with human serum and mouse serum. However, there was a slight change in the surface morphology when incubated with mouse or human serum. Similarly, a slight increase in particle size was observed after incubation with mouse or human serum (MNP-CUR-mouse serum, 136 nm; MNP-CUR-human serum, 144 nm) compared to control nanoparticles (MNP-CUR, 123 nm) (Figure 3B). The increase in particle size may be attributed to the adsorbed layers of serum proteins.50
It is imperative to assess the cytotoxicity of developed MNP-CUR for its potential anticancer application(s). The evaluation of cytotoxic activity was performed by direct observation of cell morphology as well as MTS assay. Evaluation by two different methods was conducted because some recent reports suggest that cytotoxicity assay reagents may interfere with nanoformulations.51,52 Cell image/morphology evaluation (Figure 4A) and MTS assays (Figure 4B) provide a direct qualitative and quantitative assessment of the anticancer potential of CUR/MNP-CUR formulations. When cells were treated with CUR, the viability of the cancer cells decreased sharply in a concentration (10–30 μM)-dependent manner (Figure 4A). A similar pattern was observed with MNP-CUR at all tested concentrations (Figure 4A). Interestingly, cell morphology was drastically changed and the number of cancer cells was greatly reduced even at 10 μM MNP-CUR. A very limited number of cancer cells were observed after treatment with 20 μM and 30 μM of CUR/MNP-CUR. CUR/MNP-CUR-treated cells were clearly distinguished with plasma membrane smoothing, blebbing, and loss of spindle morphology, and were almost detached, which is an apoptosis feature. Treatments with various CUR and CUR nanoformulations also have reported this apoptosis feature in the number of cancer cells.16,23 The MTS assay data demonstrated a concentration-dependent antiproliferative effect (5–40 μM) by CUR and MNP-CUR formulations (Figure 4B) as observed in the study of cellular images. The half maximal inhibitory concentration (50% cell growth inhibitory concentration) was calculated as 17.2 μM with CUR and 12.4 μM with MNP-CUR treatment. From this data, it can be concluded that although CUR release from MNP-CUR formulation was only 40%, it is as equally potent as free CUR in suppressing cell growth. Therefore, the MNP-CUR formulation can be employed as an efficient cancer therapeutic agent.
To further prove that the MNP-CUR formulation holds improved anticancer therapeutic potential, a propidium iodide stain experiment was performed. Propidium iodide specifically binds to double-stranded DNA through the plasma membrane of nonviable cells which can be detected in flow cytometry in the FL2 channel. MNP-CUR-treated cells indicate a significant propidium iodide stain over CUR-treated cells, suggesting a higher number of dead cells exist after MNP-CUR treatment (Figure A3, Appendix). This data supports the findings of MTS assay.
The authors' hypothesis is that since CUR releases slowly from MNP-CUR, this formulation may exhibit a superior anticancer activity in long term experiments such as colony formation assays. For example, magnetoplasmonic assembly of the polyethylene glycol-CUR formulation demonstrates better anticancer properties after 42–52 hours treatment compared to magnetoplasmonic assembly-CUR/ CUR treatment and the efficacy depends on the drug release pattern.49 This supports the belief that CUR nanoformulations require time to show superior effects. Therefore, the efficacy of MNP-CUR on MDA-MB-231 cancer cells was studied. A significant decrease was observed in the formation of colonies after MNP-CUR treatment compared to CUR (Figure 4C). CUR treatment gradually declined formation of colony density, ie, 4–8 μM concentration was responsible for reducing colony density from ~100% to 38% (Figure 4D) whereas 4 μM MNP-CUR treatment successfully restricted colony density to merely 16% (Figure 4D). This result clearly indicates that the increased cytotoxicity (reduced colony density) was primarily associated with active CUR release from MNP-CUR over time. A similar improved anticancer effect in a number of cancer cell lines was observed in previous studies using various CUR formulations.18–20 These studies support this enhanced anticancer potential by documenting an increase in apoptosis as demonstrated through enhanced cleaved poly(adenosine diphosphate ribose) polymerase protein with MNP or other CUR nanoformulations.18–20
The loss of mitochondrial membrane or mitochondrial dysfunction is a key indicator of intrinsic apoptosis.42 The specificity of mitochondrial stain after 24 hours treatment with CUR and MNP-CUR formulations was detected using cationic dye TMRE by fluorescence microscopy and flow cytometry. TMRE accumulates in the mitochondria when a membrane has intact potential. TMRE positive cells are an indication of viable cells with high membrane potential. Decrease in TMRE stain indicates a loss of mitochondrial membrane potential. Microscopy analysis illustrated that upon treatment with 5 μM CUR, the cells had a very similar amount of stain as control cells; however, after treatment with 10 μM CUR, the cells showed a significant decrease in stain intensity (Figure 5A). In the case of MNP-CUR, both 5 μM and 10 μM treatments showed decreased stain, indicating effective mitochondrial membrane damage and thus increased anticancer potential (Figure 5A). Similar results were observed with flow cytometry analysis. A significant decrease in TMRE stain was noticed with MNP-CUR treatment compared to CUR (Figure 5B). This may be due to higher cellular uptake of MNP-CUR, inducing greater loss in membrane potential.
Many anticancer drug treatments lead to the generation of ROS which is involved in stimulating inflammation, apoptosis, and DNA damage which can affect the signaling pathways involved in external or internal cell damage.53,54 Although CUR is a potent antioxidant with ROS scavenger characteristics, it also induces ROS production in human tumor cells.55 A MitoSOX stain was used to identify such intracellular mitochondrial ROS generation from the CUR-treated and MNP-CUR-treated cells. ROS production in the CUR-treated cells was not significantly different from control cells. However, the production of ROS was significantly increased in MNP-CUR-treated cells (Figure 6A and B). These results further demonstrate that MNP-CUR can efficiently internalize and deliver CUR in its active form and is responsible for ROS production, while free CUR barely influences the production of ROS at the tested concentrations. The obtained results corroborate similar findings reported by Mulik et al56 that CUR nanoparticles potentiate apoptosis in MDA-MB-231 cancer cells via generation of ROS. Similarly, pretreatment and cotreatment of CUR nanoparticles with other anticancer drugs also leads to an increase in ROS production.57,58
MNPs exhibit specific inherent magnetic properties that are highly important in MRI for increasing the detection limits in tissue lesions within the body.24,25,38 In addition, surface-modified MNPs avoid recognition by the reticuloendothelial system and therefore may efficiently target tumors, making them a better contrast imaging agent. In this study, the suitability of MNPs and the MNP-CUR formulation as contrast imaging agents for MRI was investigated.26,38 Figure 7A illustrates an increase of nanoparticle concentration resulting in a reduction in T2-weighted signals. Calculated transverse relaxation times from images (Figure 7A) relative to the control gel decreased with an increase in nanoparticle concentration (Figure 7B). Specifically, as the concentration increased from 10 μg iron/mL to 40 μg iron/mL, T2 relaxation times decreased from 22.4 milliseconds to 5.8 milliseconds. By plotting the transverse relaxation rate R2 (1/T2) as a function of the concentration of iron in each sample, R2 increased linearly with the concentration of iron (Figure 7C). From these data, T2 relaxivity (r2) was calculated as 7.040 s−1 μg−1 mL for MNP-CUR. The relaxivity was slightly diminished when the particles were in the presence of cancer cells (4.38 s−1 μg−1 mL) but enhanced values were observed in combination with CUR-nanoparticles. The relaxivity order detected was 7.040 s−1 μg−1 mL (MNP-CUR) > 4.38 s−1 μg−1 mL (MNP-CUR in cancer cells) > 3.40 s−1 μg−1 mL (MNP) > 2.38 s−1 μg−1 mL (MNP in cancer cells). A similar pattern was also observed in T1 relaxivity (r1): 33.21 s−1 μg−1 mL (MNP-CUR) > 18.663 s−1 μg−1 mL (MNP-CUR in cells) > 11.303 s−1 μg−1 mL (MNP) > 6.389 s−1 μg−1 mL (MNP in cancer cells). The higher relaxivity values of MNP-CUR are attributed to CUR and the nanoparticles induce greater inhomogeneity in the magnetic field than MNP. Similar improved enhanced contrast properties were observed with drug-loaded formulations due to the color(s) contributed by the drug(s), such as yellow (CUR) or red (doxorubicin).21,31,39 Previous reports support that these multilayer-coated MNPs may have superior imaging characteristics over conventional magnetic nanoformulations.40,41
Ideal magnetic field mediated drug delivery (magnetic targeted drug delivery) facilitates drug accumulation in cells possibly by passive diffusion, adsorptive transcytosis, and receptor mediation.59 This advanced drug targeting approach is increasingly utilized due to increased site specificity with fewer side effects.26 However, efficiency of magnetic field-mediated cellular delivery of drug depends on the applied external magnetic field as well as the type of MNP carrier.25,38,60,61 To determine the potential utility of the nanoformulations for possible magnetic targeting, MDA-MB-231 cancer cells treated with MNP-CUR were exposed to a neodymium external magnet for 3 hours. A detailed schematic representation of the experiment design is presented in Figure 8A. The results were evaluated comparing CUR/MNP-CUR internalization in the cancer cells. CUR (2.5–7.5 μM) exposed cells exhibited a very minute amount of CUR. MNP-CUR-treated cells demonstrated significant cellular uptake of CUR through the influence of magnetic field (Figure 8B). This effect was only minimal when there was no magnetic field applied (Figure 8B). Overall, qualitative accumulation/cellular uptake of CUR assessed by flow cytometry was clearly enhanced by magnetic field (Figure 8C) at all the concentrations tested. Thus, exposure to external magnets significantly increased the cellular uptake of MNPs predominantly into malignant cells. This study suggests that application of an external magnetic field would enhance additional CUR internalization, inducing more cellular apoptosis, thereby increasing the possible cytotoxicity of MNP-CUR.
Overall, the data show the feasibility of using an innovative nanoparticle system which combines magnetic targeting of a therapeutic agent (CUR) with both magnetic resonance and fluorescence imaging in a single system (Figure 9). This formulation can be extended to other clinically used therapeutic drugs and can be developed as antibody conjugation features.41 Targeting efficacy of these multifunctional nanoparticles specifically to triple-negative breast cancer cells can be further enhanced clinically through conjugation of monoclonal antibodies, such as antitransferrin (Figure A4, Appendix), glucose transporter 1, and folic acid.62,63
Development of safe, efficient, and biocompatible drug nanoformulations with theranostic properties is essential for clinical translation in cancer therapy. A composite MNP formulation composed of an iron oxide nanoparticle core coated with CD and pluronic polymer (F68) with CUR loading capability was developed. This formulation exhibited improved uptake in cancer cells, increased loss of mitochondrial membrane potential, and produced higher ROS, suggesting it is extremely valuable in anticancer therapeutics. Additionally, enhanced MRI properties ( compared to conventional MNPs) demonstrate this formulation has good imaging agent characteristics. More importantly, magnetic targeting of MNP-CUR facilitated an enhanced delivery of CUR to cancer cells. All together, these findings indicate that this formulation is appropriate for utilization in engineered delivery systems for theranostic applications in efficient image-directed cancer therapeutics.
The authors thank Cathy Christopherson (Sanford Research/ University of South Dakota) for editorial assistance, Robert Japs (Sanford Research/University of South Dakota), and Sara Basiaga (University of Nebraska–Lincoln) for characterization of samples. This work was partially supported by grants from Department of Defense (PC073887, PC073643), Governor's Cancer 2010, the National Institutes of Health Research Project Grant Program (RO1) (CA142736), and the Centers of Biomedical Research Excellence (P20 RR024219).
The authors report no conflicts of interest in this work.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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An Amazon and Kirkus Selection for Best Book of 2016
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The Way of the Cross or Stations of the Cross, also called as Via Crucis, means a series of images depicting the Passion of Jesus Christ- depicting the events on the day of his crucifixion, which are also associated with special prayers.
The Way of the Cross emerged as an imitation of the Jerusalem path of Via Dolorosa, which is believed to be the real path, which Jesus walked on his journey to Calvary for his crucifixion. The aim of the Stations of the Cross is for Christians to go on a spiritual journey by contemplating on the Passion of Christ.
The Stations have become one of the most popular Catholic traditions and also in several Lutheran, Methodist, and Anglican Churches.
Usually, a series of 14 images are arranged on a wall and the faithful stop by at each image and recite prayers and meditations. This can be done individually or in a group. It is typically followed on all days in the period of Lent, and finally, on Good Friday. The aim is to offer reparations for the torture and insults suffered by Jesus during the day of Crucifixion (Passion).
The placement, style and format of the stations vary. Some churches feature 14 images depicting the stations. Other minimalist designs may involve only 14 crosses. In some cases, like when the Pope leads the Stations of the Cross in the Coliseum of Rome, there might not be any images or crosses to follow, but only 14 recitations of prayers.
The fourteen Stations of the Cross are as follows:
First Station:
Jesus is condemned to die:
Jesus was most innocent. He neither could nor did commit a sin. Yet He was sentenced to death and particularly to the cruelest death by crucifixion. Pilate delivered him to the mercy of those who hated him, to preserve his friendship with Caesar.
Second Station:
Jesus is made to carry his cross:
When the cross was beheld by Jesus, he readily stretched his injured arms, embraced it willingly, kissed it with tender care and carrying the cross on his wounded shoulders; he held it willingly though he was exhausted.
Third Station:
The fall of Jesus for the first instant:
Our dear Lord, carrying the heavy cross was so burdened by its heaviness that he fell, exhausted. In fact, he was burdened by our sins and offenses. The cross was welcome, but our sins were galling.
Fourth Station:
The meeting between Jesus and his sorrowful mother:
How painful would have been it for Mother Mary, the sad mother to see her son, carrying the cross! What pangs would her tender heart may have suffered! Implore Mother Mary to assist you in your hour of death.
Fifth Station:
Jesus is helped to carry the cross by Simon of Cyrene:
It was Simon of Cyrene who was compelled to help Jesus carry the cross. Jesus accepted his assistance. Jesus is also calling you to repent of your sins and help him carry his cross. He invites you: but you decline.
Sixth Station:
The face of Jesus wiped by Veronica:
Veronica inspired by compassion and love offers her veil to wipe the disfigured face of Jesus. Jesus imprints his divine face on the veil: a great reward for her small service. What service do you perform for the wonderful blessing of the saviour?
Seventh Station:
The second fall of Jesus:
Bearing the cross's cruel and heavy burden, Jesus falls once again. But the soldiers do not allow him a moment of rest. They press him on, striking and pushing him. Jesus suffered mainly for our sins. Witnessing this passion, how can I continue to sin?
Eight Station:
The Women From Jerusalem Weep Over Jesus:
These devoted followers of Christ weep over his plight, affected by compassion. But Jesus tells them not to grieve for him, but rather to weep for themselves and their children. Most dear to the Lord is a sorrowful contrite heart crying for one's sins.
Ninth Station:
The third fall of Jesus:
At the edge of Calvary, a tired Jesus falls a third time. But his love for mankind is not diminished. How oppressive must our sins be, causing Jesus to succumb so many times!
Tenth Station:
The soldiers strip Jesus:
When Jesus arrived at Calvary, he was mercilessly stripped. It may have been very painful, tearing at his bloody skin. All garments were removed so Jesus possessed nothing. How blessed will I be, if I remove all my sinful inclinations.
Eleventh Station:
Crucifixion of Jesus:
Jesus was violently thrown upon the cross to which his feet and hands were nailed. In such excruciating pain, he did not utter anything as he bent willingly to the will of his father. He suffered in patience. This inspires me to be not fretful and complaining about my troubles.
Twelfth Station:
Raised upon the cross, Jesus dies:
Behold the crucified Jesus. He suffers for our sins. The Son of Man dies upon the cross in order to save mankind from everlasting death.
Thirteenth Station:
The body of Jesus removed from his cross and placed in the arms of his mother:
Jesus died on the cross and rested in the arms of his mother. One must persevere not to avoid the cross: who stays loyal to the cross till the last will be saved.
Fourteenth Station:
Jesus is laid in the sepulcher:
Jesus was laid in the sepulcher of a stranger. The ruler of the world did not even own a tomb of his own as he was not from this world. Do not be attached to the world, so that you may not perish with it.
These are the common Stations of the Cross. Some Christians also recognize a fifteenth station: The Resurrection of Jesus. The different stations may also vary with denominations. Every station in the Catholic Church is followed by a recitation of Stabat Mater, a prayer to Mother Mary.


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Patients work with their rehab therapists. Standing. Walking. Balancing. Healing. Taking their first steps.
See how digital signage is extending the reach of communications to patients, visitors and staff.
More than a quarter of readmissions of Medicare patients who were discharged within 30 days were actually preventable.
What are the benefits of Ethernet and the Internet Protocol for an Interactive Patient TV System?
Learn ways to win the battle of engaging hard-to-engage patients with eVideon.
Learn how movies can be used for more than just entertainment and passing time.
eVideon is the first Interactive Patient Care system to achieve Two-Way Integration with VistA and CPRS.
Why does your organization need to become a culturally competent health care organization?
Give the personal touch and improve more than just relationships


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Cash is king, but we know better than most that it can be pretty scarce. Here is some stuff that you might have lying around unused. We will use it for you!
Projector, Projector bulbs, computer paper (white and colored), printer ink cartridges, plain old writing pens, fabric and cool stuff to make puppet costumes with, alcohol and stuff to mix it with, an external hard drive, a sturdy folding table, clip lights, gels (for the lights you sillies), scrap wood, and hugs.
To contact us about a drop off or pick up please email Emily at firstname.lastname@example.com.
You will even get a thank you letter that we will hand sign ourselves! We might even spray the stationary with perfume!


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Honeycomb paper fans are so pretty. We love this Macaroon collection, these paper fans are perfect for a Birthday party, wedding or baby shower, in fact they will look gorgeous at any event or special occasion.
Colours and Sizes: each pack contains: Mint 19", Lavender 14" and Grey 12"
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This webinar was presented by Richard Perry, Chief Market Analyst at Hantec Markets
When: 6th August 2014 at 15:00 BST
During this webinar, Richard explained how to apply the RSI for trading signals activating a real-time FX trading scenario.
Following an explanation of the theory behind the RSI and a live demonstration of its practical applications Richard answered questions from the attendees.
Did you miss Top Trading Signals with the RSI webinar? Don't worry, we recorded it so if you want to know why the RSI is important in everyday trading you can access the recordered webinar here.
Top Trading Signals with RSI video
We have also prepared a PDF explaining the key concepts of Relative Strenght Index, please click on the image below.


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Pope Francis may have been elected by the Holy Spirit, but he seems made for the age of Twitter.
A case in point were his latest remarks, in which he affirmed the Catholic church's ban on artificial contraception but derided the idea that "in order to be good Catholics we have to be like rabbits" and produce litters of children.
"No," he told reporters on his flight home from the Philippines. "Responsible parenthood."
Moments earlier, Francis had signaled his approach to the vexed birth control issue when, with equally quotable verve, he said the contraception ban "does not mean that the Christian must make children in series."
He noted that during a parish visit some months ago, he even "rebuked" a woman who was pregnant again after having seven children, all delivered by Caesarean section. "But do you want to leave seven orphans?" Francis told her. "That is to tempt God!"
We say: Charlottesville reveals the weeping wound of racism. What do we, the American Catholic faith community, do next? Read the editorial.
Cue the tweets and the critiques. Rabbit breeders resented the pope's use of a derogatory cliche about libidinous bunnies, while birth control supporters said the analogy demeaned people, who should make whatever decisions they want about the number of children they have and when they have them -- or not.
Conservative Catholics and contraception opponents were also dismayed by the latest from a man who's been dubbed the "blabbermouth" pope. He appeared to criticize traditional big families (the pope said three children seemed about right) as well as undermine advocates of natural family planning (controlling birth rates without using contraception) by seeming to give aid and comfort to the church's enemies.
Francis even stumped some regular Catholics who aren't used to hearing popes speak this way, especially not about birth control, long seen as beyond debate. "As a Catholic, it's kinda shocking to hear @Pontifex say, 'Catholics must not breed like rabbits.' Really?" tweeted CNN anchor Carol Costello.
But was Francis saying anything new? Yes and no.
First, no, Francis wasn't breaking new ground. He himself told an interviewer last March that Pope Paul VI's famous encyclical reaffirming the artificial contraception ban, Humanae Vitae, was "prophetic" but said the real issue is "making sure that pastoral action takes into account that which is possible for people to do."
In other words, there is the rule, but the church must be merciful and understanding -- and use common sense. Some church conservatives saw that as more lamentable fuzziness from Francis and suggested that his predecessor, Pope Benedict XVI, a theologian with a reputation as a doctrinal hard-liner, would never have been so wobbly.
Yet in 1996, when he was Cardinal Joseph Ratzinger, Benedict said much the same thing. He told an interviewer that couples who already have several children should not be reproached if they do not want to have more, and that questions about regulating births are personal pastoral matters that "can't be projected into the abstract."
Moreover, Paul VI mentioned "responsible parenthood" in his 1968 encyclical and cited the various "physical, economic, psychological and social conditions" that could go into making decisions about when and whether to have children, stressing the primacy of individual conscience.
That approach was taken directly from a document adopted a few years earlier by the Second Vatican Council, a document that said parents should weigh a variety of "the material and the spiritual conditions" in deciding whether to have children and also stressed that "the parents themselves and no one else should ultimately make this judgment in the sight of God."
That pastoral dimension and common-sense approach got lost in the social turbulence of the 1960s and 1970s, as everyone took sides in the culture wars and tended to reduce opponents' views -- as well as their own -- to bumper-sticker slogans.
Also lost in that hubbub, however, was the fact that the church's support for regulating births even by natural means represented a major shift in Catholic teaching.
In fact, in 1930, Pope Pius XI had issued an encyclical (in response to the Anglican church's decision to OK contraception in some circumstances) saying every conjugal act must be open to the conception of children, an absolutist view that inspired decades of jokes and a memorable Monty Python song.
But in the early 1950s, the next pope, Pius XII, signaled a change when he said Catholic couples could regulate births by avoiding sex during a woman's fertile times. That was a "significant" development of church doctrine, as Italian Vatican watcher Andrea Tornielli wrote last month.
So is Francis signaling another change?
Not likely. He clearly reaffirmed the ban on artificial contraception as "prophetic," especially given declining birth rates in the West.
Yet as Francis often does, he is shifting the focus from the "rules" to the principle behind the rules. He wants Catholics to be more open to life and family and children, however many they have, and he does not think that wealthy Western countries should be telling other societies how and when to procreate.
"For the poorest people, a child is a treasure. It is true, we must also be careful here," Francis said. "But for them, a child is a treasure."
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What is your baby's favorite animal
It really varied for ours. There were times it was a struggle and we just plugged on through.
What is your baby's favorite animal?
I dunno what it'll be yet. What's YOUR favorite animal, Theresa Gould and Laura?
I like horses. With the wide array of animals we have our children now would probably say cats, though B would probably say cows and goats.
Cats and dogs .
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About Theresa Gould
Current: Chicago, Illinois
Birth: August 10
On Moms.com since: Aug 5, 2013
***Baby Team Leader Moms.com*** I have been married for over 20 years. My husband and I have eight children ages 18 down to 4 years old. We use to live in Chicago but now live in Canada. I own www.FaithandFamilyReviews.com.


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"What is happening to our health service this..."
About: Watford General Hospital Watford General Hospital Watford WD18 0HB
Posted by Anonymous
What could be improved
Not let a patient starve and saying that they will have an operation, and to let this go on for days is a disgrace and a dog would not have been treated like my daughter has been.
The government needs to go out there and do some checks on some of our appalling hospitals and see what really goes on.
Consultants should be interacting with patients and stop letting nurses be their scape goats because they think they are god
Anything else?
My daughter arrived at Watford General on monday 2nd july and after a scan and ultra sound was told she had a twisted bowel and would need an operation as it had caused a blockedge'
So on wednesday 4th they prepared her for an operation she had no food for over 24 hours and then the operation was cancelled only to be told the next day it would happen and then again it was cancelled (apparently an emergency had come in) I don't accept this as she had gone there as an emergency case'
Friday was told she was to have operation so again no food and then hours later told the operation was to be cancelled again as they had another emergency
Response from Watford General Hospital 4 years ago
Thankyou for offering feedback of your daughter's patient experience to date of writing, to which we expect would have been with Bowel Surgery, and not our Gynaecology team.
Nevertheless, we are always concerned to hear of any patients who are unhappy with the care and service they have received.
Issues of this nature are taken very seriously. The importance of a wholly positive patient experience is one of our key objectives, and the issue of communication between our staff and patients is a fundamental area we're working hard to improve in.
We know that our senior clinical colleagues will want to investigate this thoroughly on your behalf, so we can look into what went wrong for your daughter and why. Apart from giving you some answers, if would also help put things right if we can.
Therefore if you are willing to do so, please contact us directly through the Trust's Quality Assurance department at firstname.lastname@example.com . We will aim to thoroughly investigate these issues promptly, and provide you with a detailed response.
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GERMANY: Scrappage incentive helps lift Feb car market 21%
Scrappage incentives in Germany helped to lift the February car market there by 21.5%, year-on-year. However, a JD Power analyst told just-auto that manufacturer incentives on top of the government scrappage incentive also played a part in the sales surge.
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just-auto gives its paid members access to the very best automotive market coverage.
And now there's just-auto plus, our premium membership that gives you exclusive component forecast data, company profiles and extended news coverage - just for premium members.
Today I can offer you 30 days access for $1 - and that includes just-auto plus.
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SUPER BOWL XXIX; 49er Fans, Welcome To the Seifert Dynasty
By TIMOTHY W. SMITH,
Published: January 31, 1995
MIAMI, Jan. 30— Fewer than 10 hours after putting on one of the most exhilarating displays of offense in Super Bowl history, the 49ers were still giddy. And even though they could lose their offensive and defensive coordinators, it's likely that San Francisco will make a strong run for a sixth title next year.
Coach George Seifert, who said he was still numb this morning from all the excitement, was asked whether San Francisco had built a dynasty.
"I imagine it's something that perpetuates itself and maintains itself at a successful level over a period of time," he said. "Hopefully, we're looked upon as having a dynasty at this moment."
Two people who may not remain with the team are Mike Shanahan and Ray Rhodes. Shanahan, the offensive coordinator, met with the Bronco owner Pat Bowlen about the vacant head-coaching job there early today. Bowlen said he had offered him the post, but added that Shanahan had also planned to meet with the Eagles about their head-coaching vacancy this morning. Rhodes, the defensive coordinator, has been mentioned as a head coaching candidate in Philadelphia and with the Rams.
Seifert said losing Shanahan and Rhodes would be painful, but said the 49ers would not collapse. He pointed out that other coaches, including Bill Walsh, Mike Holmgren and Sam Wyche, have left the organization and the team remained successful.
The 49ers cruised through the regular season, winning 13 of 16 games, including 10 of their last 11. They steamrollered the Bears, 44-15, in the divisional round of the playoffs. They dethroned their archrivals, the hated Dallas Cowboys, 38-28, and then crushed the San Diego Chargers, 49-26, for their record fifth Super Bowl title.
After Sunday's game, the 49ers said they felt invincible. Guard Jesse Sapolu said that after San Francisco scored on its first three possessions the offensive players felt they would score every time they touched the football and were upset when drives ended with no points.
"There is no question in my mind that this is probably the best offense the N.F.L. has ever seen," Sapolu said. "People who know me, know I won't jump on any bandwagon, but if you look at our record you'll see what I'm saying is true."
For quarterback Steve Young, who threw six touchdown passes to break the Super Bowl record set by his predecessor, Joe Montana, it was a public validation of his abilities. It also lifted the burden of being reminded that Montana won four Super Bowl titles with the 49ers.
"He has always felt the pressure," said Young's agent, Leigh Steinberg. "There is an exceptional amount of pressure just being the 49ers' quarterback. It had nothing to do with his predecessor, and everything to do with the 49ers tradition."
Young said he had made the break from Montana's legacy a long time ago, but members of the news media had not.
"I did so a couple of years ago and I think you guys will too, in time," Young said. "I want my performance to stand for myself and for my teammates. It does a disservice to the team when it's talked about that way."
Young celebrated his first Super Bowl victory as the 49ers starting quarterback by receiving an intravenous solution in his hotel room Sunday night, because he was dehydrated.
He did not look any worse for the wear this morning. He joked about not knowing what to do with the new car he received for being named the game's most valuable player. He said he might give it to Shanahan as an inducement to remain with the team as offensive coordinator.
Young was asked where he thought he'd be if he had been drafted by the Bengals instead of the Buccaneers in the supplemental draft in 1984.
"I'd be with the Jets now," he joked.
Young, who began his career with the Los Angeles Express of the United States Football League, said if he could he wouldn't alter the course of anything that had led him to the Super Bowl championship and m.v.p. trophy he earned with the 49ers on Sunday night at Joe Robbie Stadium.
In terms of longevity and his future Young, 33, said he felt like he was in "the prime of my career." He spent four years as backup to Montana before getting the starting job and has not taken a beating over the last three seasons as a starter. That's because he is a great runner and flees the pocket when a sack is imminent.
Now that he has finally won a Super Bowl, Young doesn't want anyone to think that he has come to prominence all of a sudden as the quarterback of one of the most best offenses in National Football League history.
"The leap is over time," he said. "It has been a consistent building of confidence and leadership over time. It hasn't been this big coming out. It's been building over the last five years."
Photo: San Francisco's Steve Young hugged the Vince Lombardi Trophy after being named the most valuable player in Super Bowl XXIX. (Agence France-Presse)


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No kids of your own yet you know it all...?
so frustrated with my sister in law who is a close friend as well because she made some hurtful comments about my children's behavior at a recent evening out at Pizza Hut. She told her husband that her children "will never act like that" (like mine). The funny thing g is... I thought the kids were pretty well behaved. While she did not share this with me directly, the fact that she said it still hurts and is beyond irritating especially since she has no kids herself...yes everyone is entitled to their opinion but it hurts to hear someone putting you down based on the way toddlers behave. Any thoughts or advice?
Yeah. People without kids seem to know exactly how to raise them and make them behave. So many things I thought I'd never do or allow and I was humbled on all of them.
Let her say her things. Go about your day and kids.
Best thing u can do is wish kids of her own on her and them sit back and enjoy the show. ;)
Let her say her things. Go about your day and kids.
Best thing u can do is wish kids of her own on her and them sit back and enjoy the show. ;)
Just wait til she has her own....then she will see the error of her ways :)
thanks everyone! This really helped!!! :)
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About Samantha
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Birth: November 26
On Moms.com since: Feb 24, 2014
A full time working mother of 2 with one on the way. I love my 2 children more than anything and love being with them and enjoying life's little moments with them and my husband of nearly 9 years.
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Chef John
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Oct 1, 2008
The Blue Line Deli
by Natalie Mitts
The Blue Line Deli and Market is now open on the first floor of the new addition to the Tanner Building. At the Blue Line, it's New York style eating all the time. The name comes from the blue line of the subway in NYC, which runs through the financial district. BYU Dining Services decided to name the deli as such because it is located in our financial district - the Marriott School of Management in the Tanner Building. This deli features NYC cheesecake, chili, sandwiches, soups, salads, sushi, pie a la mode and Nathan's famous hot dogs. There's also a market where customers can purchase drinks, fruit and a variety of snacks, including NYC style black and white cookies. While Utah has fry sauce, the Blue Line has NYC's own sauce. Students can purchase Marriott School memorabilia and even get a copy of
The New York Times
The Wall Street Journal.
Since the setup is aimed at getting customers through fast, you can get your sandwich in a NYC minute.
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Why, hello there!
It's been a while. I know!
I'm sorry to have left you in what must have been the most suspense-filled two weeks of your life, wondering when I'll finally post another recipe {or at least I like to think that you feel this way} but life has become busy. Ever since I stepped foot into the gates of University life a month ago, I've been super swamped with work. I usually don't take busy-ness light-heartedly so when I say I have a ton of work to do then I'm definitely not exaggerating.
To give you a quick update on what's been going over here: I'm in my final year of Uni which means the countdown is on. My lectures finish in March so I'm trying to keep up with reading, assignments, etc. as good as possible while dealing with random but equally as important things like summer job applications and Grad school applications.
I have a clear vision of where I want to work next summer and which Grad school I'll attend next Autumn so I'm trying to keep my eyes on the goal. Even though it seems like both of those things are still years away.
I'm amazed at how quickly the first month back at Uni has gone by. When I wake up on Monday mornings Fridays seem sooooo far away, yet as I sit here typing this post I really don't know where the week went.
Even though I like being a busy bee and hate nothing more than boredom I really miss blogging on a regular basis. I have been taking pictures of a couple of meals but they aren't particularly styled or pretty so I'll have to think about whether the pictures will ever be published on here. Unlike any other houses or rooms I've lived at, here at Uni I seriously lack food styling equipment. I know some of you might not quite understand just how much clutter is required to create a decent looking food photograph but I can tell you that it requires more than just a plate and cutlery. Therefore, if I ever find more time to take pictures don't expect any award-winning shots.
Until then I will leave you with a recipe for a seriously satisfying, Autumn-inspired pasta dish that might knock your socks off. Adjust the amount of wine used to whatever you feel is required!
Hope you all have a lovely weekend. I know my laptop and I will have a wonderful time!
Porcini & White Wine Tagliatelle
serves 3-4
- 300-400g tagliatelle
- 2-3 tbsp. olive oil
- 2 onions, peeled and finely chopped
- 850g fresh porcini mushrooms, rinsed and finely sliced
- 250ml white wine {more if required}
- 3 cups of vegetable broth {or hot water with 1-2 bouillon cubes}
- 100ml heavy cream
- 3 tbsp. Dijon mustard
- salt and pepper
- 4 tsp. flour + water
- parsley {+ more to serve}
- parmesan optional
- Cook the pasta according to instructions on the pack. Once al dente, drain, rinse with cold water to stop the cooking process and set aside.
- In the meantime make the sauce. In a large skillet heat up the olive oil and then cook the chopped onion. Once they have softened and have become clear throw in the sliced mushrooms. Let them cook down. They will loose quite a lot of water. Stir occasionally to ensure even browning.
- When you feel that they have cooked down for most part turn up the heat to high. Then pour in the white wine and reduce almost completely. Immediately thereafter pour in the vegetable broth and stir gently. Turn down the heat to medium-high and let the sauce simmer for a short while.
- Add Dijon mustard and stir until well-combined. Season with salt and pepper.
- To thicken the sauce, stir together the flour and approx. 5 tbsp. of water to achieve a thick paste {I recommend using a glass and adding the water tablespoon by tablespoon until you have a paste-like consistency}. Turn up the heat to high and add the paste to the sauce. Stir gently and wait for the sauce to thicken.
- Fine-tune the sauce with a splash of heavy cream and season to perfection according to your taste {add whatever you feel is necessary to satisfy your taste buds}.
- At the end add some chopped parsley. Then throw the cooked pasta into the sauce, give it a good whirl and wait for the the pasta to heat up a bit.
- Serve with a generous amount of freshly grated parmesan and a bit more parsley and a glass of vino on the side.


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- hemoprotein
- noun Date: 1948 a conjugated protein (as hemoglobin or cytochrome) whose prosthetic group is a porphyrin combined with iron
New Collegiate Dictionary. 2001.
New Collegiate Dictionary. 2001.
Hemoprotein — A hemoprotein (also haemoprotein), or heme protein, is a metalloprotein containing a heme prosthetic group, either covalently or noncovalently bound to the protein itself. The iron in the heme is capable of undergoing oxidation and reduction… … Wikipedia
hemoprotein — hemoproteinas statusas T sritis chemija apibrėžtis Baltymas, turintis hemą ir pernešantis deguonį. atitikmenys: angl. hemoprotein rus. гемопротеин … Chemijos terminų aiškinamasis žodynas
hemoprotein — Protein linked to a metal porphyrin compound ( e.g., cytochromes, myoglobin, catalase). * * * he·mo·pro·tein or chiefly Brit hae·mo·pro·tein prō .tēn, prōt ē ən n a conjugated protein (as hemoglobin or cytochrome) whose prosthetic group is a… … Medical dictionary
hemoprotein — he·mo·protein … English syllables
hemoprotein — noun a conjugated protein linked to a compound of iron and porphyrin • Syn: ↑haemoprotein • Hypernyms: ↑conjugated protein, ↑compound protein • Hyponyms: ↑hemoglobin, ↑haemoglobin, ↑H … Useful english dictionary
NADPH—hemoprotein reductase — Identifiers EC number 126.96.36.199 CAS number 9023 03 4 … Wikipedia
NADPH-hemoprotein reductase — In enzymology, a NADPH hemoprotein reductase (EC number|184.108.40.206) is an enzyme that catalyzes the chemical reaction:NADPH + H+ + n oxidized hemoprotein ightleftharpoons NADP+ + n reduced hemoproteinThe 3 substrates of this enzyme are NADPH, H+,… … Wikipedia
NADPH–hemoprotein reductase — NADPH–he·mo·pro·tein re·duc·tase (heґmo pro″tēn re dukґtās) [EC 126.96.36.199] an enzyme of the oxidoreductase class that catalyzes the transfer of electrons to cytochromes via oxidation of NADPH. It is a flavoprotein (FMN, FAD)… … Medical dictionary
Cytochrome P450 reductase — P450 (cytochrome) oxidoreductase PDB rendering based on 1amo … Wikipedia
cytochrome — A class of hemoprotein whose principal biologic function is electron and/or hydrogen transport by virtue of a reversible valency change of the heme iron. Cytochromes are classified in four groups (a, b, c, and d) according to spectrochemical… … Medical dictionary


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1.3 Overview
This section provides a high-level overview of the operation of the Remote Desktop Protocol: Audio Input Redirection Virtual Channel Extension. The purpose of this protocol is to transfer audio data from a Remote Desktop Protocol (RDP) client to a Remote Desktop Protocol (RDP) server, hereinafter referred to as client and server, respectively. For example, an application running on a server can request to record audio data. This data will be transferred from the client to the server, allowing a server application to record from an audio device installed on the client.
The protocol is divided into three main sequences:
Initialization sequence: The server and client exchange versions and audio formats, and begin recording.
Data transfer sequence: The client sends audio data to the server.
Format change sequence: The server requests a new audio format, and the client confirms this request.