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Amazon won't likely make a bid for Electronic Arts.
Shares of Electronic Arts surged Friday after USA Today reported that Amazon plans to make a bid to acquire the video game maker.
Amazon is not expected to make a bid to acquire Electronic Arts, sources told CNBC's David Faber on Friday.
EA shares closed up 3.5% Friday after USA Today reported that Amazon would announce Friday that it plans to make a bid for the video game publisher known for hit titles such as FIFA, Madden and Apex Legends.
Faber dismissed the report on CNBC's Squawk Box. "There's nothing to it," he said, citing sources.
An Amazon spokesman said the company does not comment on rumors or speculation. A spokesman for EA declined to comment.
If Amazon seriously pursues EA, it could help bolster the retail giant's gaming presence. Amazon already owns Twitch, a streaming service popular with gamers, has launched its own cloud gaming service called Luna and offers perks for gamers as part of its Prime loyalty program.
Amazon has been very active on the M&A front. Earlier this month, Amazon announced that it was acquiring iRobot, best known for its Roomba robot vacuum cleaner, for $1.7 billion. This announcement came just two weeks after it announced it was buying boutique chain One Medical for $3.9 billion.
The gaming industry has seen some consolidation this year. In January, Microsoft announced it would buy Activision Blizzard in a $68.7 billion deal. And in May, Take-Two Interactive announced it would acquire Zynga for $11 billion.


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WOODLAND HILLS, Calif., Nov. 16, 2012 /PRNewswire/ -- GrowLife, Inc. (PHOT), an industry-leading consortium of high-end gardening and horticulture brands focused on merchandising the "picks and shovels" of the growing "green rush", today announced that it was featured in a recent article written by Bruce Kennedy on MSN Money. The article highlighted how changes in state marijuana laws are encouraging some cannabis-related companies to go public, including GrowLife Inc., Hemp Inc. (HEMP), Medical Marijuana Inc. (MJNA).
Sterling Scott, GrowLife, Inc. CEO, was quoted in the article as saying, "It was almost unthinkable 10 years ago that you would have legitimate, fully reporting to the SEC companies that were in the nature of pure plays, with positions in the medical marijuana industry." Now, Mr. Scott estimates that there are about 10 cannabis-related companies being traded over-the-counter, including GrowLife, Inc. – an established company that sells equipment and expendables.
The full article on MSN Money can be read here:
GrowLife's consortium of companies actively participate in all channels of commerce for distribution and sales, including operating on-line and retail outlets for direct consumer sales and the supply of specialty GrowLife company products to wholesale and retail channels. And, while supporting progressive normalization of cannabis laws, the company does not grow, sell or distribute any substances that violate U.S. law or the Federal Controlled Substances Act.
About GrowLife, Inc.
GrowLife, Inc. (PHOT) (formerly Phototron Holding, Inc.) (www.growlifeinc.com) (PHOT) is a company with core holdings in innovative technology-based products and services for the indoor gardening industry and specialty markets. These brands include Stealth Grow, a producer of grow room automation equipment and hi-powered LED grow light products for indoor horticulture (www.sgsensors.com and www.stealthgrow.com), Greners.com, the online hydroponics superstore (www.greners.com) and Phototron, producer of hydroponic grow containers, which are designed to grow vegetables, herbs, flowers and fruits in any environment (www.phototron.com).
For more information about our public holding company, please visit www.growlifeinc.com.
Cautionary Language Concerning Forward-Looking Statements
Information set forth in this press release contains financial estimates and other forward-looking statements that are subject to risks and uncertainties, and actual results might differ materially. A discussion of factors that may affect future results is contained in GrowLife's filings with the Securities and Exchange Commission. In addition, all industry products are subject to additional uncertainty, including the risks of delay, cancellation and poor critical or financial reception. GrowLife disclaims any obligation to update and revise statements contained in this news release based on new information or otherwise.
SOURCE GrowLife, Inc.

domain: news


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- The NY Times has a lengthy review of Leapfrog's Fly Pentop edutainment gadget, which is appearing on all of those "hot toys for Christmas lists" that gets parents in a tizzy. Aimed at the hard-to-please tween set, the loquacious Pentop reads text aloud from special paper (only sold by Leapfrog, natch) and uses program modules that are — you guessed it — sold separately at $25-$35 a pop. Unless I'm misunderstanding the article, Leapfrog spent $100 designing the little bugger so I guess it's okay to fleece parents with impunity, if only because it really does seem to make learning fun for the kids who use it. [NY Times]
- If you prick call center operators, do they not bleed? More and more American customers are going ballistic on the guys and gals who field customer service calls, which, as you ve probably noticed, are increasingly being outsourced to other countries. Unfortunately, many of these heated (one-sided) discussions get very, very ugly these days as customers are pelting the poor souls with cruel racial insults and profanity that would make a Ku Klux Klan member cringe. Chill out everyone!
[San Francisco Chronicle]
- Thanks to slick manufacturing wizardry, Texas Instruments, Intel, and other semiconductor companies have managed to shoehorn most of the main functions of a cellphone onto a single chip, paving the way for cheaper phones. [WSJ (reg)]
- Sony and NEC ink a deal to co-manufacture their DVD and CD drives. One big problem they need to work out: Sony's the leader of the Blu-Ray gang and NEC's allegiances are with HD-DVD. [Chicago Tribune].
- WebTV lives! But Microsoft is calling the sequel MSN TV2 Internet & Media Player (ugh-they should've stuck with the old name; at least Microsoft kept the WebTV.com domain) and despite some major upgrades, still only appears to be fit for tech-phobic senior citizens looking for a simple way to email their loved ones that they've fallen and can't get up. [Baltimore Sun].
We may earn a commission from links on this page.


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That 70-site guide, that has long since gone out of produce (but nonetheless retains a hallowed put on my display), was among the guides that sparked my interest in mind strength and hypnosis. Should you try weight loss hypnosis that will help you lose weight , then you might be to attaining your weight loss objectives possibly nearer. you are helped by weight loss hypnosis one of the most challenging to obtain rid down, in smashing previous habits, and building fresh good eating up habits. Weight-loss hypnosis also encourages healthy food to be prepared by one and consume the meals, that will be good for you and the one that is acceptable for reaching your weight reduction goals. Your perspective changes towards weight-management and raises on your own-confidence.
Effective weight reduction depends upon your perspective towards it and then you can do just this if you would like to utilize hypnosis to really get your brain round weight loss' irritating matter. Hypnosis hyped up promises or can nevertheless be abused from stop smoking hypnosis the hypnotist or online marketer generating fantastic plus it can be abused by those that do not actually offer it a good try.
Many individuals realize that when they follow a fat loss hypnosis system that exercise becomes something and more enjoyable they really want to do. They are some of the useful methods trance can help people. Those who have abandoned on diets that are past find that they believe it is easier to stick after fat loss trance to them, and in addition since they start to feel the diets could work that the performance of the diets increases.

domain: health


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AUTISM, MENTAL HEALTH AND SUICIDE
I was glad to be able to participate in a debate on Autism, Mental Health and Suicide last week.
Recent research shows that 66% of those diagnosed with autism have considered suicide.
Clearly there is a need for more investment in mental health from the UK Government particularly for those living with autism.
Whilst the Scottish Government has done important work in developing and creating a Scottish Strategy for Autism, funding squeezes from the UK Government creates ongoing challenges.
Young people living with autism are 28 times more likely to consider suicide than other young people so clearly the challenge is to provide sufficient support for those living with autism.
Indeed, a study carried out in Sweden concluded that suicide is the leading cause of premature death in people with autism.
Please click on the link below to hear my contribution to the debate


By Onome Amuge & Zainab Iwayemi
A total of 73,000 people in the workforce of Lagos State are fully under insurance coverage as Government House, Alausa, Ikeja says it has paid N2 billion to cover both life and non-life insurance premiums for the workers.
The premiums paid by the state are up-to-date as they cover the current year 2021, disclosed Rabiu Olowo, the state’s commissioner for finance, at the Lagos State Civil Service Insurance Week, which kicked off on Monday, August 23.
Olowo highlighted the importance of insurance and the role the government plays to protect the entire workforce of 73,000 workers in the state, encouraging the masses on the need to insure their lives, properties and valuables.
Noting that the Lagos State government has continued to insure its staff with regular payment of premiums, while insurers on their part, continue to deliver payment of the death benefits to deceased beneficiaries, the commissioner said insurance is not optional as it is one of the means through which the state government manages risk in the state.
According to Olowo, the insurance plan by the Babajie Sanwo-Olu led administration is in line with commitments to ensuring adequate care for its workforce while serving and after retirement.
He therefore added that insurance is compulsory in Lagos state for life and non-life insurance assets to protect against future events drawing from the COVID-19 pandemic and the civil unrest experienced in Lagos, hence, the reason for further sensitisation and education on insurance and also the maiden edition of the Lagos State Civil Service Insurance Week.
“Insurance is not an issue of optionality or so. We have life and non-life insurance and that is very compulsory. Presently, we have over 73,000 employees on our database from the ministries and all these employees are covered under life insurance,” Olowo said.
The commissioner also explained that the insurance department in the ministry of finance provides professional risk advisory to the state and also coordinates the various insurance policies and packages of the state.
“The task of ensuring that vehicles, buildings, electronics, computers, firefighting equipment, other assets, as well as the welfare package of the entire workforce, is the responsibility of the insurance department,” he stressed.
Olowo bemoaned that many workers in the state are not fully aware of the existence of a professional insurance department domiciled in the ministry of finance and, as such, have not been maximizing the crucial role being performed by the department.
To this end, he assured that the state government would take advantage of this year’s insurance week ceremony to explore the various advantages and benefits of insurance to the entire workforce. He also noted that the state government has identified the need for more sensitisation and education on insurance, and how civil servants and the entire populace can key into the abundant opportunities in insurance.
Olowo added that the four-day insurance week, which would span between 23rd August to 26th August will serve as an avenue to educate participants on the ways civil servants and the entire populace can utilise the abundant opportunities in insurance.
He said, “We have seen government property assets destroyed and in some cases injuries to our staff and associates. We have enjoyed the real essence and value proposition of insurance, and the best we can do is to further our commitment to ensuring that our insurance activities are strengthened. That is why we are doing this sensitisation and advocacy and we can see that it pays to pay insurance premiums and we will continue to do so into the future.”
Adding that there is a need to institute the culture of insurance consciousness among government officials and the citizenry as a whole, the commissioner maintained that having appropriate insurance cover could not be overemphasised, being the acceptable practice across the globe whilst he noted that the current administration is setting pace in the area of insurance to encourage residence embrace the initiative.
“In other words, the cases of calling on the government at all times for compensation over some natural disasters or unfortunate incidences can become a thing of the past if we have necessary insurance cover for ourselves, businesses and property.
“By this, the resources at the disposal of the government can be channelled towards other developmental projects, instead of paying compensations that ordinarily could have been undertaken by insurance companies,” he said.


Atlanta producer Mike-WiLL Made-It released his first official album today, and it features a star-studded list of rap contributors. Ransom 2 was originally supposed to be a mixtape released in January 2016, but the platinum-selling producer postponed the release so he could feature Gucci Mane, a long-time friend and collaborator. Over that time he built up more impressive collaborations and turned the project into a full album.
The wait was absolutely worth it though, as that song now brings together Kendrick Lamar, Gucci Mane, and Rae Sremmurd on the same track. The 17-track project feature some of the biggest names in hip-hop including Pharrell, Lil Wayne, Big Sean, 2 Chainz, and many more. Make WiLL has made himself a staple in hip-hop over the past few years producing tons of artists' hits and brings out this star-studded crew for his own project.
Mike Will brings on Rae Sremmurd, who he discovered and signed to his label Ear Drummers. He actually even convinced them to make their artist name the backwards spelling of his label. Mike Will has had a strong involvement with Rae Sremmurds releases and has word closely with tons of high profile artists like Kanye, Future, Rihanna, Big Sean, Pusha T, Gucci Mane and many more.
Mike WiLL’s production ranges from minimal trap beats to hard-hitting bass lines, and he makes everyone sound natural over them. The entire project brings more of the ATL sound that’s dominating the charts right now, so check out these songs before you hear them all over the radio this summer. Enjoy!


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See-through Mask, Offsite Project, Glasgow (2004)
With Clare Stephenson and Craig Mulholland
With: Craig Mulholland, Alex Pollard, Clare Stephenson
As co-curator of Switchspace, a voluntary organization that transforms vacant Glasgow properties into temporary exhibition spaces, Sorcha Dallas has been ardently committed to the Glasgow visual art community since the late 1990s. For the inaugural annual programme of her new gallery, Dallas has chosen to represent and promote the practices of six artists drawn from this milieu and entourage. Each could be seen to share a common interest in an opulent and perhaps romantic approach to their furtive and esoteric subject matter. Most adopt an ambivalent (and often camp) engagement with conventional media such as painting, sculpture and drawing. A fluid variety in the handling combines to produce multivalent installations and environments. The significance of their work falls equally on the haptic and the optic, on making and taking, on making things and making things up.
Craig Mulholland – Born in Glasgow in 1969, Mulholland completed his BA in Drawing and Painting at Glasgow School of Art in 1991. MulHolland has exhibited regularly in group and solo shows in Glasgow, Edinburgh and London. He currently works with a complex configuration of media, including oil painting, meticulous digital animation and phantasmagorical tableaux. Recent installations have explored abbreviated connections between music and the visual arts, focusing in particular on the mythological interplay between American modernists and bluegrass guitar maestros.
Neil Mulholland 2004
Preview Saturday 29th May, 7-9PM Until 3rd July Gallery hours 1-5pm Thursday-Saturday or by appointment
- Off site project featuring Craig Mulholland, Alex Pollard and Clare Stephenson*

domain: artistic


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From time to time, I get empty stomach or grumbling feeling below the sternum. No pain/ everything else is alright. My appetite is good, bowel movements are normal. The sensation can last several days, subside for days or weeks and then reappear. Ideas?
Welcome to icliniq.com.
I understand your concerns.
From what you mentioned, most probably you have dyspepsia which is a problem due to abnormal gastric acid secretion.
I suggest a trial of Pantoprazole 20 mg tablet once a day before breakfast for a month.
Eat simple meals.
Have Maalox (Magnesium hydroxide and Aluminium hydroxide) sachet after main meals.
After one month if symptoms are still present Helicobacter pylori test will be required.
For more information consult a general medicine physician online --> https://www.icliniq.com/ask-a-doctor-online/general-medicine-physician
Was this answer helpful?|
Same symptoms doesn’t mean you have the same problem. Consult a doctor now!
Heartburn (acidity) ... The stomach environment is extremely acidic ... Read fullI am getting diarrhea after taking Kremil-S and Omeprazole. What to do?
.. reviewed your history as well as the medication profile you just shared. (attachment removed to protect patient identity). So the part of the problem is secondary to the medicine you are taking such as diarrhea which tends to occur secondary to ... Read fullI often get a grumbling feeling in my stomach. What could it be?
.. your concerns. From what you mentioned, most probably you have dyspepsia which is a problem due to abnormal gastric acid secretion. I suggest a trial of Pantoprazole 20 mg tablet once a day before breakfast for a month. Eat simple meals. Have ... Read full
Ask your health query to a doctor online?Ask a General Medicine Physician Now

domain: medical


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Disc one has spoken word poems written and performed by Léonie Scott-Matthews. Disc two has the poems set to music performed by various musicians including David.
Poetry has always been Léonie’s first love. Her poet and musician father encouraged her to write and compose, while her actress and drama teacher mother taught her to recite and perform. After training at the Royal Academy of Music and Drama and then winning the Poetry Society’s Gold Medal for Verse Speaking, in 1968 Léonie founded Pentameter’s, a poetry based theatre in Hampstead. Now in 2021 she is presenting her own poetry, music, stories and plays.
All songs and spoken word recorded and produced by Matthew Kitchiner
David Nevin & Godfrey Old have a new album out
Moon at Night is an alternative open evening held at The Pentameter Theatre in Hampstead NW3 – originally founded by actress, poet and musician Léonie Scott-Matthews in 1968.
David regularly attends this open evening performing his own songs with Léonie’s musician
partner Godfrey Old. Having developed a working relationship with Godfrey he has collaborated on a selection of cover songs that have both influenced their musical life.
David Nevin – Guitar and vocal
Godfrey Old – Harmonica
Produced by Matthew Kitchiner
Mystery of Love
Here’s a video for the track ‘Desperation Drive’ from my upcoming album ‘Fantasy’
You can listen to the full length album ‘Strangelove’ on Davids Bandcamp page here

domain: entertainment


Quiet Ocean Surveys Begin
Now that COVID-19 restrictions have been lifted in Grand Cayman to allow scuba diving, CCMI researchers have been conducting coral reef surveys. The ocean has had almost no direct human impact for over 4 months, thus the Quiet Ocean study is looking to track the impact of ocean use by humans as restrictions are eased. We believe the lack of human interaction may have had a positive impact on fish populations in traditionally heavily used areas, such as George Town Harbor. To investigate this, scientists have completed a series of initial fish surveys at 5 sites along the west end, 3 of which are in the harbor and in direct proximity to where the cruise ships anchor. We will compare these populations to data from 2018 when CCMI conducted AGRRA surveys on Grand Cayman (as well as Little Cayman and Cayman Brac), and we will also repeat the surveys every 2-3 months as restrictions begin to ease and impact increases. In this way, we hope to have quantifiable data showing the progression of impact that tourism has on Cayman’s coral reefs, and specifically the local fish populations.


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Swedish stainless steel producer Sandvik group reported a 38pc fall in operating profits in Q2 to SEK2,858mn ($314.6mn) driven by a steep decline in revenue and order intake due to the pandemic.
Sandvik’s chief executive officer, Stefan Widing said that the company faced its biggest drop in orders amid declines in the automotive and aerospace sectors. Mining activity was also affected by mine closures.
In the second quarter, the company posted revenue of SEK20.2bn ($2.2bn), a decline of 20pc over the prior-year period. EBITDA stood at SEK 3.4bn ($368mn), down by 66pc over Q2 2019.
Sandvik Mining and Rock Technology
Orders dropped by 10pc to $1bn, with a majority of them for underground mining tools from Australia, the Middle East, and South Africa.
Sandvik Machining Tools
Order intake declined by 35pc to $750mn driven by the plummeting production in the automotive and aerospace sectors across Europe and North America. Chinese imports rose marginally in Q2. Revenue dropped 32pc to $798mn.
Sandvik Materials Technology
Order intake dropped by a third to $257mn. The company noted an operating loss of $9mn. A large order for nuclear housing material was booked from China leading to positive year-on-year growth for the company’s Asia division. Tube products suffered due to weak oil and gas markets. Demand for heating materials and heating systems also saw a decline.
In the past year, Sandvik divested 70pc of its Varel division and used these proceeds to acquire Newtrax in Canada, Thermaltek, Melin Tool and Summerill Tube in the US and Quimmico Centro Technologies in Mexico to strengthen its hold in the North American market for its core products.
(SEK = $0.11)


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Th17 cells are a distinct subset of T cells that have been found to produce interleukin 17 (IL-17), and differ in function from the other T cell subsets including Th1, Th2, and regulatory T cells. Th17 cells have emerged as a central culprit in overzealous inflammatory immune responses associated with many autoimmune disorders. In this method we purify T lymphocytes from the spleen and lymph nodes of C57BL/6 mice, and stimulate purified CD4+ T cells under control and Th17-inducing environments. The Th17-inducing environment includes stimulation in the presence of anti-CD3 and anti-CD28 antibodies, IL-6, and TGF-β. After incubation for at least 72 hours and for up to five days at 37 °C, cells are subsequently analyzed for the capability to produce IL-17 through flow cytometry, qPCR, and ELISAs. Th17 differentiated CD4+CD25- T cells can be utilized to further elucidate the role that Th17 cells play in the onset and progression of autoimmunity and host defense. Moreover, Th17 differentiation of CD4+CD25- lymphocytes from distinct murine knockout/disease models can contribute to our understanding of cell fate plasticity.
19 Related JoVE Articles!
Imaging Denatured Collagen Strands In vivo and Ex vivo via Photo-triggered Hybridization of Caged Collagen Mimetic Peptides
Institutions: University of Utah, Johns Hopkins University School of Medicine, Johns Hopkins University.
Collagen is a major structural component of the extracellular matrix that supports tissue formation and maintenance. Although collagen remodeling is an integral part of normal tissue renewal, excessive amount of remodeling activity is involved in tumors, arthritis, and many other pathological conditions. During collagen remodeling, the triple helical structure of collagen molecules is disrupted by proteases in the extracellular environment. In addition, collagens present in many histological tissue samples are partially denatured by the fixation and preservation processes. Therefore, these denatured collagen strands can serve as effective targets for biological imaging. We previously developed a caged collagen mimetic peptide (CMP) that can be photo-triggered to hybridize with denatured collagen strands by forming triple helical structure, which is unique to collagens. The overall goals of this procedure are i)
to image denatured collagen strands resulting from normal remodeling activities in vivo
, and ii)
to visualize collagens in ex vivo
tissue sections using the photo-triggered caged CMPs. To achieve effective hybridization and successful in vivo
and ex vivo
imaging, fluorescently labeled caged CMPs are either photo-activated immediately before intravenous injection, or are directly activated on tissue sections. Normal skeletal collagen remolding in nude mice and collagens in prefixed mouse cornea tissue sections are imaged in this procedure. The imaging method based on the CMP-collagen hybridization technology presented here could lead to deeper understanding of the tissue remodeling process, as well as allow development of new diagnostics for diseases associated with high collagen remodeling activity.
Bioengineering, Issue 83, collagen remodeling, triple helix, near infrared fluorescence, bioimaging, tissue staining
A 3D System for Culturing Human Articular Chondrocytes in Synovial Fluid
Institutions: Tufts University School of Medicine, Tufts Medical Center.
Cartilage destruction is a central pathological feature of osteoarthritis, a leading cause of disability in the US. Cartilage in the adult does not regenerate very efficiently in vivo
; and as a result, osteoarthritis leads to irreversible cartilage loss and is accompanied by chronic pain and immobility 1,2
. Cartilage tissue engineering offers promising potential to regenerate and restore tissue function. This technology typically involves seeding chondrocytes into natural or synthetic scaffolds and culturing the resulting 3D construct in a balanced medium over a period of time with a goal of engineering a biochemically and biomechanically mature tissue that can be transplanted into a defect site in vivo 3-6
. Achieving an optimal condition for chondrocyte growth and matrix deposition is essential for the success of cartilage tissue engineering.
In the native joint cavity, cartilage at the articular surface of the bone is bathed in synovial fluid. This clear and viscous fluid provides nutrients to the avascular articular cartilage and contains growth factors, cytokines and enzymes that are important for chondrocyte metabolism 7,8
. Furthermore, synovial fluid facilitates low-friction movement between cartilaginous surfaces mainly through secreting two key components, hyaluronan and lubricin 9 10
. In contrast, tissue engineered cartilage is most often cultured in artificial media. While these media are likely able to provide more defined conditions for studying chondrocyte metabolism, synovial fluid most accurately reflects the natural environment of which articular chondrocytes reside in.
Indeed, synovial fluid has the advantage of being easy to obtain and store, and can often be regularly replenished by the body. Several groups have supplemented the culture medium with synovial fluid in growing human, bovine, rabbit and dog chondrocytes, but mostly used only low levels of synovial fluid (below 20%) 11-25
. While chicken, horse and human chondrocytes have been cultured in the medium with higher percentage of synovial fluid, these culture systems were two-dimensional 26-28
. Here we present our method of culturing human articular chondrocytes in a 3D system with a high percentage of synovial fluid (up to 100%) over a period of 21 days. In doing so, we overcame a major hurdle presented by the high viscosity of the synovial fluid. This system provides the possibility of studying human chondrocytes in synovial fluid in a 3D setting, which can be further combined with two other important factors (oxygen tension and mechanical loading) 29,30
that constitute the natural environment for cartilage to mimic the natural milieu for cartilage growth. Furthermore, This system may also be used for assaying synovial fluid activity on chondrocytes and provide a platform for developing cartilage regeneration technologies and therapeutic options for arthritis.
Cellular Biology, Issue 59, Chondrocytes, articular, human, synovial fluid, alginate bead, 3D culture
In Vivo Optical Imaging of Brain Tumors and Arthritis Using Fluorescent SapC-DOPS Nanovesicles
Institutions: University of Cincinnati College of Medicine, University of Cincinnati College of Medicine, University of Cincinnati College of Medicine, University of Cincinnati College of Medicine, The Ohio State University Medical Center, The Ohio State University Medical Center.
We describe a multi-angle rotational optical imaging (MAROI) system for in vivo
monitoring of physiopathological processes labeled with a fluorescent marker. Mouse models (brain tumor and arthritis) were used to evaluate the usefulness of this method. Saposin C (SapC)-dioleoylphosphatidylserine (DOPS) nanovesicles tagged with CellVue Maroon (CVM) fluorophore were administered intravenously. Animals were then placed in the rotational holder (MARS) of the in vivo
imaging system. Images were acquired in 10° steps over 380°. A rectangular region of interest (ROI) was placed across the full image width at the model disease site. Within the ROI, and for every image, mean fluorescence intensity was computed after background subtraction. In the mouse models studied, the labeled nanovesicles were taken up in both the orthotopic and transgenic brain tumors, and in the arthritic sites (toes and ankles). Curve analysis of the multi angle image ROIs determined the angle with the highest signal. Thus, the optimal angle for imaging each disease site was characterized. The MAROI method applied to imaging of fluorescent compounds is a noninvasive, economical, and precise tool for in vivo
quantitative analysis of the disease states in the described mouse models.
Medicine, Issue 87, Saposin C (SapC), Dioleoylphosphatidylserine (DOPS), Brain tumor, Arthritis, Fluorophore, Fluorescence, Optical imaging, Multi-angle rotational optical imaging (MAROI)
Establishment of a Surgically-induced Model in Mice to Investigate the Protective Role of Progranulin in Osteoarthritis
Institutions: NYU Hospital for Joint Diseases, New York University Medical Center.
Destabilization of medial meniscus (DMM) model is an important tool for studying the pathophysiological roles of numerous arthritis associated molecules in the pathogenesis of osteoarthritis (OA) in vivo
. However, the detailed, especially the visualized protocol for establishing this complicated model in mice, is not available. Herein we took advantage of wildtype and progranulin (PGRN)-/- mice as examples to introduce a protocol for inducing DMM model in mice, and compared the onset of OA following establishment of this surgically induced model. The operations performed on mice were either sham operation, which just opened joint capsule, or DMM operation, which cut the menisco-tibial ligament and caused destabilization of medial meniscus. Osteoarthritis severity was evaluated using histological assay (e.g.
Safranin O staining), expressions of OA-associated genes, degradation of cartilage extracellular matrix molecules, and osteophyte formation. DMM operation successfully induced OA initiation and progression in both wildtype and PGRN-/- mice, and loss of PGNR growth factor led to a more severe OA phenotype in this surgically induced model.
Bioengineering, Issue 84, Mouse, Cartilage, Surgery, Osteoarthritis, degenerative arthritis, progranulin, destabilization of medial meniscus (DMM)
DNBS/TNBS Colitis Models: Providing Insights Into Inflammatory Bowel Disease and Effects of Dietary Fat
Institutions: BC Children's Hospital.
Inflammatory Bowel Diseases (IBD), including Crohn's Disease and Ulcerative Colitis, have long been associated with a genetic basis, and more recently host immune responses to microbial and environmental agents. Dinitrobenzene sulfonic acid (DNBS)-induced colitis allows one to study the pathogenesis of IBD associated environmental triggers such as stress and diet, the effects of potential therapies, and the mechanisms underlying intestinal inflammation and mucosal injury. In this paper, we investigated the effects of dietary n-3 and n-6 fatty acids on the colonic mucosal inflammatory response to DNBS-induced colitis in rats. All rats were fed identical diets with the exception of different types of fatty acids [safflower oil (SO), canola oil (CO), or fish oil (FO)] for three weeks prior to exposure to intrarectal DNBS. Control rats given intrarectal ethanol continued gaining weight over the 5 day study, whereas, DNBS-treated rats fed lipid diets all lost weight with FO and CO fed rats demonstrating significant weight loss by 48 hr and rats fed SO by 72 hr. Weight gain resumed after 72 hr post DNBS, and by 5 days post DNBS, the FO group had a higher body weight than SO or CO groups. Colonic sections collected 5 days post DNBS-treatment showed focal ulceration, crypt destruction, goblet cell depletion, and mucosal infiltration of both acute and chronic inflammatory cells that differed in severity among diet groups. The SO fed group showed the most severe damage followed by the CO, and FO fed groups that showed the mildest degree of tissue injury. Similarly, colonic myeloperoxidase (MPO) activity, a marker of neutrophil activity was significantly higher in SO followed by CO fed rats, with FO fed rats having significantly lower MPO activity. These results demonstrate the use of DNBS-induced colitis, as outlined in this protocol, to determine the impact of diet in the pathogenesis of IBD.
Medicine, Issue 84, Chemical colitis, Inflammatory Bowel Disease, intra rectal administration, intestinal inflammation, transmural inflammation, myeloperoxidase activity
A Mouse Model for Pathogen-induced Chronic Inflammation at Local and Systemic Sites
Institutions: Boston University School of Medicine, Boston University School of Medicine.
Chronic inflammation is a major driver of pathological tissue damage and a unifying characteristic of many chronic diseases in humans including neoplastic, autoimmune, and chronic inflammatory diseases. Emerging evidence implicates pathogen-induced chronic inflammation in the development and progression of chronic diseases with a wide variety of clinical manifestations. Due to the complex and multifactorial etiology of chronic disease, designing experiments for proof of causality and the establishment of mechanistic links is nearly impossible in humans. An advantage of using animal models is that both genetic and environmental factors that may influence the course of a particular disease can be controlled. Thus, designing relevant animal models of infection represents a key step in identifying host and pathogen specific mechanisms that contribute to chronic inflammation.
Here we describe a mouse model of pathogen-induced chronic inflammation at local and systemic sites following infection with the oral pathogen Porphyromonas gingivalis
, a bacterium closely associated with human periodontal disease. Oral infection of specific-pathogen free mice induces a local inflammatory response resulting in destruction of tooth supporting alveolar bone, a hallmark of periodontal disease. In an established mouse model of atherosclerosis, infection with P. gingivalis
accelerates inflammatory plaque deposition within the aortic sinus and innominate artery, accompanied by activation of the vascular endothelium, an increased immune cell infiltrate, and elevated expression of inflammatory mediators within lesions. We detail methodologies for the assessment of inflammation at local and systemic sites. The use of transgenic mice and defined bacterial mutants makes this model particularly suitable for identifying both host and microbial factors involved in the initiation, progression, and outcome of disease. Additionally, the model can be used to screen for novel therapeutic strategies, including vaccination and pharmacological intervention.
Immunology, Issue 90,
Pathogen-Induced Chronic Inflammation; Porphyromonas gingivalis; Oral Bone Loss; Periodontal Disease; Atherosclerosis; Chronic Inflammation; Host-Pathogen Interaction; microCT; MRI
Analysis of Cell Migration within a Three-dimensional Collagen Matrix
Institutions: Witten/Herdecke University.
The ability to migrate is a hallmark of various cell types and plays a crucial role in several physiological processes, including embryonic development, wound healing, and immune responses. However, cell migration is also a key mechanism in cancer enabling these cancer cells to detach from the primary tumor to start metastatic spreading. Within the past years various cell migration assays have been developed to analyze the migratory behavior of different cell types. Because the locomotory behavior of cells markedly differs between a two-dimensional (2D) and three-dimensional (3D) environment it can be assumed that the analysis of the migration of cells that are embedded within a 3D environment would yield in more significant cell migration data. The advantage of the described 3D collagen matrix migration assay is that cells are embedded within a physiological 3D network of collagen fibers representing the major component of the extracellular matrix. Due to time-lapse video microscopy real cell migration is measured allowing the determination of several migration parameters as well as their alterations in response to pro-migratory factors or inhibitors. Various cell types could be analyzed using this technique, including lymphocytes/leukocytes, stem cells, and tumor cells. Likewise, also cell clusters or spheroids could be embedded within the collagen matrix concomitant with analysis of the emigration of single cells from the cell cluster/ spheroid into the collagen lattice. We conclude that the 3D collagen matrix migration assay is a versatile method to analyze the migration of cells within a physiological-like 3D environment.
Bioengineering, Issue 92, cell migration, 3D collagen matrix, cell tracking
Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog
Institutions: Lehigh University.
Post-translational modifications may lead to altered protein functional states by increasing the covalent variations on the side chains of many protein substrates. The histone tails represent one of the most heavily modified stretches within all human proteins. Peptidyl-arginine deiminase 4 (PAD4) has been shown to convert arginine residues into the non-genetically encoded citrulline residue. Few assays described to date have been operationally facile with satisfactory sensitivity. Thus, the lack of adequate assays has likely contributed to the absence of potent non-covalent PAD4 inhibitors. Herein a novel fluorescence-based assay that allows for the monitoring of PAD4 activity is described. A pro-fluorescent substrate analog was designed to link PAD4 enzymatic activity to fluorescence liberation upon the addition of the protease trypsin. It was shown that the assay is compatible with high-throughput screening conditions and has a strong signal-to-noise ratio. Furthermore, the assay can also be performed with crude cell lysates containing over-expressed PAD4.
Chemistry, Issue 93, PAD4, PADI4, citrullination, arginine, post-translational modification, HTS, assay, fluorescence, citrulline
In vitro Coculture Assay to Assess Pathogen Induced Neutrophil Trans-epithelial Migration
Institutions: Harvard Medical School, MGH for Children, Massachusetts General Hospital.
Mucosal surfaces serve as protective barriers against pathogenic organisms. Innate immune responses are activated upon sensing pathogen leading to the infiltration of tissues with migrating inflammatory cells, primarily neutrophils. This process has the potential to be destructive to tissues if excessive or held in an unresolved state. Cocultured in vitro
models can be utilized to study the unique molecular mechanisms involved in pathogen induced neutrophil trans-epithelial migration. This type of model provides versatility in experimental design with opportunity for controlled manipulation of the pathogen, epithelial barrier, or neutrophil. Pathogenic infection of the apical surface of polarized epithelial monolayers grown on permeable transwell filters instigates physiologically relevant basolateral to apical trans-epithelial migration of neutrophils applied to the basolateral surface. The in vitro
model described herein demonstrates the multiple steps necessary for demonstrating neutrophil migration across a polarized lung epithelial monolayer that has been infected with pathogenic P. aeruginosa
(PAO1). Seeding and culturing of permeable transwells with human derived lung epithelial cells is described, along with isolation of neutrophils from whole human blood and culturing of PAO1 and nonpathogenic K12 E. coli
(MC1000). The emigrational process and quantitative analysis of successfully migrated neutrophils that have been mobilized in response to pathogenic infection is shown with representative data, including positive and negative controls. This in vitro
model system can be manipulated and applied to other mucosal surfaces. Inflammatory responses that involve excessive neutrophil infiltration can be destructive to host tissues and can occur in the absence of pathogenic infections. A better understanding of the molecular mechanisms that promote neutrophil trans-epithelial migration through experimental manipulation of the in vitro
coculture assay system described herein has significant potential to identify novel therapeutic targets for a range of mucosal infectious as well as inflammatory diseases.
Infection, Issue 83, Cellular Biology, Epithelium, Neutrophils, Pseudomonas aeruginosa, Respiratory Tract Diseases, Neutrophils, epithelial barriers, pathogens, transmigration
Measuring Ascending Aortic Stiffness In Vivo in Mice Using Ultrasound
Institutions: Johns Hopkins University, Johns Hopkins University, Johns Hopkins University, Macquarie University.
We present a protocol for measuring in vivo
aortic stiffness in mice using high-resolution ultrasound imaging. Aortic diameter is measured by ultrasound and aortic blood pressure is measured invasively with a solid-state pressure catheter. Blood pressure is raised then lowered incrementally by intravenous infusion of vasoactive drugs phenylephrine and sodium nitroprusside. Aortic diameter is measured for each pressure step to characterize the pressure-diameter relationship of the ascending aorta. Stiffness indices derived from the pressure-diameter relationship can be calculated from the data collected. Calculation of arterial compliance is described in this protocol.
This technique can be used to investigate mechanisms underlying increased aortic stiffness associated with cardiovascular disease and aging. The technique produces a physiologically relevant measure of stiffness compared to ex vivo
approaches because physiological influences on aortic stiffness are incorporated in the measurement. The primary limitation of this technique is the measurement error introduced from the movement of the aorta during the cardiac cycle. This motion can be compensated by adjusting the location of the probe with the aortic movement as well as making multiple measurements of the aortic pressure-diameter relationship and expanding the experimental group size.
Medicine, Issue 94, Aortic stiffness, ultrasound, in vivo, aortic compliance, elastic modulus, mouse model, cardiovascular disease
Meal Duration as a Measure of Orofacial Nociceptive Responses in Rodents
Institutions: Texas A&M University Baylor College of Dentistry.
A lengthening in meal duration can be used to measure an increase in orofacial mechanical hyperalgesia having similarities to the guarding behavior of humans with orofacial pain. To measure meal duration unrestrained rats are continuously kept in sound attenuated, computerized feeding modules for days to weeks to record feeding behavior. These sound-attenuated chambers are equipped with chow pellet dispensers. The dispenser has a pellet trough with a photobeam placed at the bottom of the trough and when a rodent removes a pellet from the feeder trough this beam is no longer blocked, signaling the computer to drop another pellet. The computer records the date and time when the pellets were taken from the trough and from this data the experimenter can calculate the meal parameters. When calculating meal parameters a meal was defined based on previous work and was set at 10 min (in other words when the animal does not eat for 10 min that would be the end of the animal's meal) also the minimum meal size was set at 3 pellets. The meal duration, meal number, food intake, meal size and inter-meal interval can then be calculated by the software for any time period that the operator desires. Of the feeding parameters that can be calculated meal duration has been shown to be a continuous noninvasive biological marker of orofacial nociception in male rats and mice and female rats. Meal duration measurements are quantitative, require no training or animal manipulation, require cortical participation, and do not compete with other experimentally induced behaviors. These factors distinguish this assay from other operant or reflex methods for recording orofacial nociception.
Behavior, Issue 83, Pain, rat, nociception, myofacial, orofacial, tooth, temporomandibular joint (TMJ)
Feeding of Ticks on Animals for Transmission and Xenodiagnosis in Lyme Disease Research
Institutions: Tulane University Health Sciences Center.
Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi,
occurs by the attachment and blood feeding of Ixodes
species ticks on mammalian hosts. In nature, this zoonotic bacterial pathogen may use a variety of reservoir hosts, but the white-footed mouse (Peromyscus leucopus
) is the primary reservoir for larval and nymphal ticks in North America. Humans are incidental hosts most frequently infected with B. burgdorferi
by the bite of ticks in the nymphal stage. B. burgdorferi
adapts to its hosts throughout the enzootic cycle, so the ability to explore the functions of these spirochetes and their effects on mammalian hosts requires the use of tick feeding. In addition, the technique of xenodiagnosis (using the natural vector for detection and recovery of an infectious agent) has been useful in studies of cryptic infection. In order to obtain nymphal ticks that harbor B. burgdorferi
, ticks are fed live spirochetes in culture through capillary tubes. Two animal models, mice and nonhuman primates, are most commonly used for Lyme disease studies involving tick feeding. We demonstrate the methods by which these ticks can be fed upon, and recovered from animals for either infection or xenodiagnosis.
Infection, Issue 78, Medicine, Immunology, Infectious Diseases, Biomedical Engineering, Primates, Muridae, Ticks, Borrelia, Borrelia Infections, Ixodes, ticks, Lyme disease, xenodiagnosis, Borrelia, burgdorferi, mice, nonhuman primates, animal model
Protein WISDOM: A Workbench for In silico De novo Design of BioMolecules
Institutions: Princeton University.
The aim of de novo
protein design is to find the amino acid sequences that will fold into a desired 3-dimensional structure with improvements in specific properties, such as binding affinity, agonist or antagonist behavior, or stability, relative to the native sequence. Protein design lies at the center of current advances drug design and discovery. Not only does protein design provide predictions for potentially useful drug targets, but it also enhances our understanding of the protein folding process and protein-protein interactions. Experimental methods such as directed evolution have shown success in protein design. However, such methods are restricted by the limited sequence space that can be searched tractably. In contrast, computational design strategies allow for the screening of a much larger set of sequences covering a wide variety of properties and functionality. We have developed a range of computational de novo
protein design methods capable of tackling several important areas of protein design. These include the design of monomeric proteins for increased stability and complexes for increased binding affinity.
To disseminate these methods for broader use we present Protein WISDOM (https://www.proteinwisdom.org), a tool that provides automated methods for a variety of protein design problems. Structural templates are submitted to initialize the design process. The first stage of design is an optimization sequence selection stage that aims at improving stability through minimization of potential energy in the sequence space. Selected sequences are then run through a fold specificity stage and a binding affinity stage. A rank-ordered list of the sequences for each step of the process, along with relevant designed structures, provides the user with a comprehensive quantitative assessment of the design. Here we provide the details of each design method, as well as several notable experimental successes attained through the use of the methods.
Genetics, Issue 77, Molecular Biology, Bioengineering, Biochemistry, Biomedical Engineering, Chemical Engineering, Computational Biology, Genomics, Proteomics, Protein, Protein Binding, Computational Biology, Drug Design, optimization (mathematics), Amino Acids, Peptides, and Proteins, De novo protein and peptide design, Drug design, In silico sequence selection, Optimization, Fold specificity, Binding affinity, sequencing
In vivo Macrophage Imaging Using MR Targeted Contrast Agent for Longitudinal Evaluation of Septic Arthritis
Institutions: University Hospital of Strasbourg, University of Strasbourg, University Hospital of Strasbourg.
Macrophages are key-cells in the initiation, the development and the regulation of the inflammatory response to bacterial infection. Macrophages are intensively and increasingly recruited in septic joints from the early phases of infection and the infiltration is supposed to regress once efficient removal of the pathogens is obtained. The ability to identify in vivo
macrophage activity in an infected joint can therefore provide two main applications: early detection of acute synovitis and monitoring of therapy.
noninvasive detection of macrophages can be performed with magnetic resonance imaging using iron nanoparticles such as ultrasmall superparamagnetic iron oxide (USPIO). After intravascular or intraarticular administration, USPIO are specifically phagocytized by activated macrophages, and, due to their magnetic properties, induce signal changes in tissues presenting macrophage infiltration. A quantitative evaluation of the infiltrate is feasible, as the area with signal loss (number of dark pixels) observed on gradient echo MR images after particles injection is correlated with the amount of iron within the tissue and therefore reflects the number of USPIO-loaded cells.
We present here a protocol to perform macrophage imaging using USPIO-enhanced MR imaging in an animal model of septic arthritis, allowing an initial and longitudinal in vivo
noninvasive evaluation of macrophages infiltration and an assessment of therapy action.
Medicine, Issue 80, Biomedical Engineering, Anatomy, Physiology, Molecular Biology, Diagnostic Imaging, Musculoskeletal System, Bacterial Infections and Mycoses, Macrophage, MR imaging, infection, arthritis, USPIO, imaging, clinical techniques
Granulocyte-dependent Autoantibody-induced Skin Blistering
Institutions: University of Freiburg , Kepler High School Freiburg, University of Freiburg .
Autoimmune phenomena occur in healthy individuals, but when self-tolerance fails, the autoimmune response may result in specific pathology. According to Witebsky's postulates, one of the criteria in diagnosing a disease as autoimmune is the reproduction of the disease in experimental animals by the passive transfer of autoantibodies. For epidermolysis bullosa acquisita (EBA), a prototypic organ-specific autoimmune disease of skin and mucous membranes, several experimental models were recently established. In the animal model described in our present work, purified IgG antibodies against a stretch of 200 amino acids (aa 757-967) of collagen VII are injected repeatedly into mice reproducing the blistering phenotype as well as the histo- and immunopathological features characteristic to human EBA 1
. Full-blown widespread disease is usually seen 5-6 days after the first injection and the extent of the disease correlates with the dose of the administered collagen VII-specific IgG. The tissue damage (blister formation) in the experimental EBA is depending on the recruitment and activation of granulocytes by tissue-bound autoantibodies 2,-4
. Therefore, this model allows for the dissection of the granulocyte-dependent inflammatory pathway involved in the autoimmune tissue damage, as the model reproduces only the T cell-independent phase of the efferent autoimmune response. Furthermore, its value is underlined by a number of studies demonstrating the blister-inducing potential of autoantibodies in vivo
and investigating the mechanism of the blister formation in EBA 1,3,-6
. Finally, this model will greatly facilitate the development of new anti-inflammatory therapies in autoantibody-induced diseases. Overall, the passive transfer animal model of EBA is an accessible and instructive disease model and will help researchers to analyze not only EBA pathogenesis but to answer fundamental biologically and clinically essential autoimmunity questions.
Immunology, Issue 68, Medicine, Physiology, Anatomy, Dermatology, autoimmunity, collagen VII, inflammation, extracellular matrix, Fc receptor, complement, granulocyte, antibody
Antibody Transfection into Neurons as a Tool to Study Disease Pathogenesis
Institutions: Veterans Administration Medical Center, Memphis, TN, University of Tennessee Health Science Center, Memphis, TN, University of Tennessee Health Science Center, Memphis, TN.
Antibodies provide the ability to gain novel insight into various events taking place in living systems. The ability to produce highly specific antibodies to target proteins has allowed for very precise biological questions to be addressed. Importantly, antibodies have been implicated in the pathogenesis of a number of human diseases including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), paraneoplastic syndromes, multiple sclerosis (MS) and human T-lymphotropic virus type 1 (HTLV-1) associated myelopathy/tropical spastic paraparesis (HAM/TSP) 1-9
. How antibodies cause disease is an area of ongoing investigation, and data suggests that interactions between antibodies and various intracellular molecules results in inflammation, altered cellular messaging, and apoptosis 10
. It has been shown that patients with MS and HAM/TSP produce autoantibodies to the intracellular RNA binding protein heterogeneous ribonuclear protein A1 (hnRNP A1) 3, 5-7, 9, 11
. Recent data indicate that antibodies to both intra-neuronal and surface antigens are pathogenic 3, 5-9, 11
. Thus, a procedure that allows for the study of intracellular antibody:protein interactions would lend great insight into disease pathogenesis.
Genes are commonly transfected into primary cells and cell lines in culture, however transfection of antibodies into cells has been hindered by alteration of antibody structure or poor transfection efficiency 12
. Other methods of transfection include antibody transfection based on cationic liposomes (consisting of DOTAP/DOPE) and polyethylenimines (PEI); both of which resulted in a ten-fold decrease in antibody transfection compared to controls 12
. The method performed in our study is similar to cationic lipid-mediated methods and uses a lipid-based mechanism to form non-covalent complexes with the antibodies through electrostatic and hydrophobic interactions 13
. We utilized Ab-DeliverIN reagent, which is a lipid formulation capable of capturing antibodies through non-covalent electrostatic and hydrophobic interactions and delivering them inside cells. Thus chemical and genetic couplings are not necessary for delivery of functional antibodies into living cells. This method has enabled us to perform various antibody tracing and protein localization experiments, as well as the analyses of the molecular consequences of intracellular antibody:protein interactions 9
In this protocol, we will show how to transfect antibodies into neurons rapidly, reproducibly and with a high degree of transfection efficiency. As an example, we will use anti-hnRNP A1 and anti-IgG antibodies. For easy quantification of transfection efficiency we used anti-hnRNP A1 antibodies labelled with Atto-550-NHS and FITC-labeled IgG. Atto550 NHS is a new label with high molecular absorbtion and quantum yield. Excitation source and fluorescent filters for Atto550 are similar to Cy3 (Ex. 556 Em. 578). In addition, Atto550 has high photostability. FITC-labeled IgG were used as a control to show that this method is versatile and not dye dependent. This approach and the data that is generated will assist in understanding of the role that antibodies to intracellular target antigens might play in the pathogenesis of human diseases.
Neuroscience, Issue 67, Medicine, Molecular Biology, Immunology, Transfection, antibodies, neuron, immunocytochemistry, fluorescent microscopy, autoimmunity
Mammary Transplantation of Stromal Cells and Carcinoma Cells in C57BL/6J Mice
Institutions: University of Kansas Medical Center.
The influence of stromal cells, including fibroblasts on mammary tumor progression has been well documented through the use of mouse models, in particular through transplantation of stromal cells and epithelial cells in the mammary gland of mice. Current transplantation models often involve the use of immunocompromised mice due to the different genetic backgrounds of stromal cells and epithelial cells. Extracellular matrices are often used to embed the two different cell types for consistent cell-cell interactions, but involve the use of Matrigel or rat tail collagen, which are immunogenic substrates. The lack of functional T cells from immunocompromised mice prevents accurate assessment of stromal cells on mammary tumor progression in vivo
, with important implications on drug development and efficacy. Moreover, immunocompromised mice are costly, hard to breed and require special care conditions. To overcome these obstacles, we have developed an approach to orthotopically transplant stromal cell and epithelial cells into mice from the same genetic background to induce consistent tumor formation. This system involves harvesting normal, carcinoma associated fibroblasts, PyVmT mammary carcinoma cells and collagen from donor C57BL/6J mice. The cells are then embedded in collagen and transplanted in the inguinal mammary glands of female C57BL/6J mice. Transplantation of PyVmT cells alone form palpable tumors 30-40 days post transplantation. Endpoint analysis at 60 days indicates that co-transplantation with fibroblasts enhances mammary tumor growth compared to PyVmT cells transplanted alone. While cells and matrix from C57BL/6J mice were used in these studies, the isolation of cells and matrix and transplantation approach may be applied towards mice from different genetic backgrounds demonstrating versatility. In summary, this system may be used to investigate molecular interactions between stromal cells and epithelial cells, and overcomes critical limitations in immunocompromised mouse models.
Medicine, Issue 54, transplantation, mammary, fibroblast, PyVmT carcinoma, collagen type-I , tumor
Reverse Total Shoulder Arthroplasty
Institutions: Case Western Reserve University.
Reverse total shoulder arthroplasty was initially approved for use in rotator cuff arthropathy and well as chronic pseudoparalysis without arthritis in patients who were not appropriate for tendon transfer reconstructions. Traditional surgical options for these patients were limited and functional results were sub-optimal and at times catastrophic. The use of reverse shoulder arthroplasty has been found to effectively restore these patients function and relieve symptoms associated with their disease. The procedure can be done through two approaches, the deltopectoral or the superolateral. Complication rates associated with the use of the prosthesis have ranged from 8-60% with more recent reports trending lower as experienced is gained. Salvage options for a failed reverse shoulder prosthesis are limited and often have significant associated disability. Indications for the use of this prosthesis continue to be evaluated including its use for revision arthroplasty, proximal humeral fracture and tumor. Careful patient selection is essential because of the significant risks associated with the procedure.
Medicine, Issue 53, Reverse, Total, Shoulder, Arthroplasty, Rotator Cuff, Arthropathy, Arthritis, Glenoid, Humerus, Fracture
Non-invasive Imaging of Leukocyte Homing and Migration in vivo
Institutions: Washington University in St. Louis, NIH - National Institute of Health.
Two-photon (2P) microscopy is a high resolution imaging technique that has been broadly adapted by biologists. The value of 2P microscopy is that it provides rich spatiotemporal information regarding cell behaviors within intact tissues and in live mice. Leukocyte recruitment plays a significant role in host defense against infection and when unchecked, can contribute to inflammatory and autoimmune diseases. Studying leukocyte recruitment in vivo
is technically challenging since cells are moving rapidly within vessels located deep within light scattering tissues. To date, most intravital imaging studies require surgical preparation to expose the blood vessels and tissues. To avoid the tissue damage and inflammation induced by surgery itself, here, we describe a non-invasive single-cell imaging approach that can be used to study leukocyte trafficking in the mouse footpad and phalanges. We discuss the technical aspects of our 2P imaging preparation and walk the reader through a typical experiment from initial set up to execution and data collection.
Immunology, issue 46, Two-photon microscopy, leukocytes, homing, inflammation


text:
Beautiful 1970s Thai Silk Floral Dress
1970s Thai Silk Dress
Availability: 1 in stock
- Era 1970s
- Bust 36"
- Waist 26" - 37" (elasticated)
- Hips Free
- Label Indra
- Fabric Silk
- Condition Excellent
This item is in excellent vintage condition.
- Best fit UK 12/14
The best fit is a recommendation only. Please refer to other measurements for accurate fitting.
Absolutely dreamy Thai silk dress in the most beautiful floral print topped off with amazing leg of mutton sleeves, covered buttons, original belt and pleat details. Exceptionally made and with an elasticated waist it is super comfortable too.
Model is 5’10” and a size UK 10/12
Sustainable and unique clothing
Ashwell & Co take inspiration from the past to create a unique vintage shopping experience, with sustainability, slow fashion and individuality at the heart of what we do.
From 1930s bias cut gowns to 1940s tea dresses; 1960s mod suits to 1970s dagger collar shirts, our carefully curated collection of vintage fashion caters for a wide variety of tastes, sizes and budgets.
Each vintage piece has been handpicked for our rails and sourced exclusively within the UK by our co-founder Kate.
From iconic design pieces such as Ossie Clark, Yves Saint Laurent and Lanvin, to everyday essentials, each piece of clothing is chosen as the best example of its era, whilst also keeping an emphasis on wearability and current trends.

domain: design


text:
Fair food is by definition outrageous, and this year’s OC Fair Food (at the Orange County Fair) is no exception. Huge portions, crazy food combinations, and deep-fried everything is the rule. Of course we (and probably most people) wouldn’t eat like this every day. But we’re still fascinated by fair food. Each year some of […]
Tag Archives | fair food
Hi! I'm Jeanne Fratello - a writer, reporter, and food enthusiast who is on a mission to write great stories about food and where your food comes from. You can read more about me here.
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apples bacon bananas bread breakfast cake carrots cauliflower cheese chicken chocolate cookies dessert eggs farm fresh fruits and veggies gluten-free holidays kale Melissa's Produce Melissas Netflix olive oil onions organic peppers pie pineapple pizza potatoes real food salad spinach strawberries strawberry StreamTeam Stream Team summer Thanksgiving tomatoes vegan vegetarian veggies watermelon zucchini

domain: food


text:
We are Theatre Passe Muraille, a theatre company located in Toronto — currently developing and producing new plays. We strive to articulate a distinct Canadian voice that reflects the complexity of our intercultural society.
Hey, you’re part of it!
“Experience the taste of the Woking Phoenix!”
An epic exploration of two decades of a family’s survival. Woking Phoenix tells the story of three siblings, their mother, and their restaurant as they seek to find belonging within small-town Ontario. An intergenerational Chinese love story about creating community and the food that reminds us of home.
The sophomore edition of our presentation festival #BeyondTO bringing works to our historic red doors from outside Toronto. This year we welcome Soliloquio (Argentina) and Camille: The Story (Québec) to share intimate stories from the breadth of the human experience.
Explore TPM’s 23.24 Season!
Collage by Emily Jung
Pictured from left to right: Donna-Michelle St. Bernard, Charlie Petch, Gloria Mok and family, Luke Reece, Dario Dvon, Sky-Ravinn, Courage Bacchus, Selecta Sarah.
A thrilling new season of eclectic, moving, and boundary pushing works, full of surprises. This season boasts four productions developed at TPM through our Buzz program. From supporting VR conference attendees to a de-escalation workshop and everything in-between, we hope you will join us through our red doors, as we continue our journey of bringing you some of the most exciting artists and work in town!


text:
Submitted by Matt Lindsay on Sep122016
Little Miami Scenic Trail in Springfield
Submitted by Matt Lindsay on Nov232015
This edition of Trail Talk - the MVBT quarterly news letter - has a little of everything.
Submitted by Matt Lindsay on Nov132014
Submitted by Matt Lindsay on Jul072014
Trail damage caused by heavy storms in June resulted in a trail closure at Sycamore Creek, just north of downtown Miamisburg. As of July 7, 2014, this trail repair project has been completed by the Miami Conservancy District and the trail has been re-opened for use by the general public. You can read more about the issue here.
Many thanks to MCD for a fast project to restore the trail, and to you for your patience.

domain: geography


text:
In a firm appeal to Libyan factions on Wednesday, the United Nations Support Mission in Libya (UNSMIL) urged a “political settlement” to resolve ongoing discord, concerning the recently ratified election laws.
The legal frameworks, affirmed by the Tobruk-based Parliament, continue to spark debate with the High Council of State (HCS) in Tripoli, illustrating a palpable divide.
On 2 October, the election laws received unilateral approval from the Parliament. However, resistance from the HCS soon emerged, emphasising a divergence from texts previously agreed upon by the 6+6 Joint Committee in Bouznika, Morocco.
In a statement, UNSMIL highlighted that despite forming a basis for elections, the laws have “contentious issues that necessitate addressing and resolution through a political settlement.” The statement urges “commitment in good faith” from all parties, particularly key leaders, to bring these laws to fruition.
UNSMIL emphasised that their team has completed the initial technical review of the “Presidential Election Law No. (28) of 2023” and the “Parliamentary Election Law No. (27) of 2023.”
The UN outlined several matters indicative of mistrust amongst Libyan parties, notably encompassing four primary points enshrined in the laws.
The stipulations involve the mandatory execution of a second round for Presidential elections, the linkage between Presidential and Parliamentary elections, the establishment of a unified government to transition to elections, and the comprehensive inclusion of all Libyans, including women and diverse cultural constituents.
UNSMIL underscored these issues as inherently political, necessitating a “national settlement to propel the country towards elections.” It called for key factions to demonstrate good faith, engage in constructive dialogue, and deliver a definitive resolution to these enduring issues.
Libya has grappled with significant political unrest and instability since the overthrow and killing of its longtime leader, Muammar Gaddafi in 2011. In the subsequent power vacuum, the country plunged into chaos with various militias and factions vying for control.

domain: law


text:
Pretzels, raisins and almonds are coated with melted chocolate and formed into clusters for a delicious easy-to-make snack.
What You Need
Original recipe yields 10 servings
1 cup miniature pretzel twists
1/2 cup seedless raisins
1/3 cup PLANTERS Slivered Almonds
2 pkg. (4 oz. each) BAKER'S Semi-Sweet Chocolate, broken into pieces, melted
Add To Shopping List
Let's Make It
Combine pretzels, raisins and almonds in large bowl. Add chocolate; toss until pretzel mixture is evenly coated.
Drop by rounded tablespoonfuls onto wax paper-covered baking sheet. Let stand 1 hour at room temperature or until chocolate is firm.
Remove from wax paper. Store in tightly covered container at room temperature.
Substitute dried cranberries for the raisins.
Calories From Fat
% Daily Value*
Total Fat 9g
Saturated Fat 4.5g
Trans Fat 0g
Total Carbohydrates 23g
Dietary Fibers 2g
* Percent Daily Values are based on a 2,000 calorie diet.
10 servings, 2 clusters each
* Nutrition information is estimated based on the ingredients and cooking instructions as described in each recipe and is intended to be used for informational purposes only. Please note that nutrition details may vary based on methods of preparation, origin and freshness of ingredients used.


Some of the most important decisions in laying out a shop involve the placement of basic machines, such as the table saw and jointer. Deciding where to put them depends in part on the scale of woodworking you do, which determines the clearance area around these machines if you need to handle long stock or large sheet goods. You’ll also want to consider work flow to avoid an excess amount of running around the shop to accomplish tasks, as well as how to power your machines without turning your shop into a snakepit of extension cords.
I have never worked with my bench up against the wall, because I like to work from all 4 sides of my bench. I do like my machines pushed up against the wall and a large open space in the middle of the shop to facilitate easier movement. I have a Minimax combination machine so by necessity my tablesaw and jointer/planer are bundled together but I would actually prefer they not be (like a Felder machine that breaks apart). I would never put my tablesaw central in my shop, always at one end with a clear entry and egress for the sliding table, space permitting. Putting a tablesaw in the middle always struck me as one thing I’d always have to walk around and/or fight the temptation to stack stuff on it.
I do not rip on the table saw as a rule, to prevent kick back that periodically occurs when natural wood pinches the blade, turning the wood into a missile. I bought a fairly powerful saw, so this is one place where a lighter saw would be adequate. Essentially, I use the table saw for ripping sheet goods, cutting dados, tenons, and cutting small parts to length – all of which can be done with a 1.5 HP saw. I ended up with a King 3 HP, three-belt drive, 10" table saw. The castings are true, and the King Tru-rip fence reminds me of the Biesmeyer fence used on the Canadian General saws. The model is KC-11FX, and it can be purchased for less than half of the price of other, similar saws. On this purchase, I went with the suggestion of Jeff at Brettwood Machinery – he was right; a very good value saw that runs smooth, and has a decent fence.
Jointer: I use my jointer a lot. When preparing rough lumber it sees as much action as the planer. As a matter of fact, almost every piece of lumber in my shop gets surfaced on the wide face to straighten things out before it even heads to the planer. Without the jointer, my life would just be a crooked, twisty mess of painful attempts to make things seem straight.
Commonly used woodworking tools included axes, adzes, chisels, pull saws, and bow drills. Mortise and tenon joints are attested from the earliest Predynastic period. These joints were strengthened using pegs, dowels and leather or cord lashings. Animal glue came to be used only in the New Kingdom period. Ancient Egyptians invented the art of veneering and used varnishes for finishing, though the composition of these varnishes is unknown. Although different native acacias were used, as was the wood from the local sycamore and tamarisk trees, deforestation in the Nile valley resulted in the need for the importation of wood, notably cedar, but also Aleppo pine, boxwood and oak, starting from the Second Dynasty.
I know this seems really simple and you may already know how to read a tape measure so just hear me out on this one! Often times with woodworking you need to make exact measurements and cuts and it’s rarely pretty even numbers like 15 inches or 15 1/2 inches. It’s usually like 15 5/8 inches or 15 9/16 inches. So, really knowing how to read a tape measure in its entirety is important. And I’ve created a quick post on how to read a tape measure the easy way along with a helpful free printable.
In each case, we shuffled the bench, jointer, table saw, and band saw across to the top of the stairs, and then tied a rope around each to act as a safety while sliding the machines down the strapping on the stairs. Yeah, the table saw hit the wall, and the promise of a good mud and paint job saved my bacon. The rope worked well, and we were able to get everything down the stairs nice and slowly without any major issues.
Lacking a jointer? Use reader Court Kites’ awesome tip to create perfectly matched glue joints on wavy or bowed board edges. Lay the boards on a ﬂat surface, then clamp them across the middle with a bar clamp. Lay two 8-in. long by 1-3/4 in. wide scrap boards across each end and screw them in with four 1-1/4 in. long screws, two per board. Keep the screws well away from your future cutting line!
I really appreciate the article. I am always looking for a way to improve my shop and its layout. I currently have a 34′ x 28′ garage which I park two vehicles and still have some room for a table saw, large bench and various smaller benches and cabinets. All of my large power tools are on casters so I can roll them out when I move the cars outside. Right now I am considering keeping my little truck outside permanently and set the shop up more in a fixed layout. This would make working in the shop more enjoyable and much easier to do more complex projects that I currently take on. Maybe this spring. Thanks for the advice.
Cedars are strong, aromatic softwoods that are capable of enduring outdoor elements, the most common of which is the Western Red Cedar. Western Red Cedar can sustain wet environments without succumbing to rot, and as a result is commonly used for outdoor projects such as patios, outdoor furniture, and building exteriors. This wood can be easily found at most home centers for a moderate price.
Ultimately, your workshop should be a place that inspires you to be creative and to do your best work. When you take the time to plan the space for maximum efficiency and comfort, you’ll love spending time there — and that means you’ll get more done, whether it’s fixing up a car or building a new piece of furniture. No matter how you use your workshop, you should enjoy the time you get to work with your hands and make something brand new. So what are you waiting for? Get started designing your perfect workshop today.
Short scraps of hardwood are too good to throw away but hard to store neatly. So I bought a 4-ft. tube form made for concrete footings, cut it in half (the cardboard-like material cuts easily) and set the tubes on end. I tack the tubes to a wall or a bench leg so they don’t fall over. With the wood scraps stored upright, it’s easy to find a piece just the right length. Tube forms are available in various diameters for $5 and up at home centers. — Bill Wells. Here’s another brilliant use for these concrete forms.
A little different from the others, the Wood Shop is geared more towards the “self-helper.” Take the three-hour safety class, available every Saturday, to earn your qualification card. You can then utilize the shop to work on your own projects. A wonderful variety of wood working equipment is available for customer usage for a very reasonable fee. Be sure to check out the available classes where you can learn to make pens, boxes, decorative cutting boards and even rocking chairs. We also produce custom order items to include coin holders, specialized plaques and shadow boxes.
Clamping mitered edges can be a real hassle because they never seems to line up correctly. The easiest way that I’ve found to get around this process is to use painter’s tape as clamps. First set the pieces so that the outer edges are facing up and tape them edge-to-edge. The flip the pieces over so the beveled edges are facing up and glue them together. Complete the process by taping the last two edges together and let sit until completed. The tape removes easily and the glue won’t attach to the tape, making sanding and finishing very simple. Try this tip with this clever project!
Cutting sandpaper is a quick way to dull your scissors or utility knife blade. Instead, I fastened a hacksaw blade to the edge of my workbench. I slipped a washer behind the blade at each of the mounting holes so a sheet of sandpaper to easily slides in behind the blade. I fold the paper where I want to cut, just as a reference. — Kim Boley. Try some of these storage solutions!
To make sure you account for any existing obstructions, it’s a good idea to make a measured drawing of your workshop area on graph paper, noting existing furniture, built-ins and large items on the plan as you go. For example, a common one-car garage workshop layout has built-in tool cabinets and shelves around the perimeter where they won’t impede parking, but you may also have to get clever about folding work tables and saw horses that you can set up as needed in the middle of the garage when you’re working on a project (and cars are parked outside).


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Review by JustinW303
Reviewed: 02/22/03 | Updated: 02/22/03
Mediocre, bland, average. Notice the theme here?
We all knew it would happen. With the overwhelming popularity of anything remotely related to Harry Potter, everyone knew there had to be a video game somewhere down the line. Fans of the series embraced the idea, genuinely excited by just the thought of being able to control Harry and live out his first year at the legendary wizarding school. Others, though, cringed at the mere mention of yet another game based on a license. What could it bring to the table that would make it anything worth playing? How accurately could EA emulate the atmosphere of the books? The final product isn’t anything amazing or groundbreaking in the slightest, but anyone interested in the series ought to give it a try.
Being as the game is based on a best-selling book, you’d naturally expect it to rely heavily on the story elements of that book. With this game, such an assumption would be proven wrong within the first few minutes of playing. While the game does make a vague attempt to stay true to the book, anyone playing this game without first reading the book may be a little confused as the game progresses, and for just reason. Of course, some things had to be tweaked a bit for the game to be anything worth playing, but it sometimes strays a little too far away from the source material. I’m all for adding things to a game like this if it adds depth without changing the story too much, but sometimes I think EA tried to warp it a little more than needed, and ended up distorting the story to the point where I wouldn’t have recognized it had the Harry Potter name not been on it. Things are added, removed, and switched around in an effort to better involve the player, assuming the person has a background knowledge of the books. I would strongly suggest reading the book before playing the game so you not only know what the game is about, but so that you derive a little more enjoyment from the game as a whole. This game was obviously made with the fan in mind, not the average gamer.
With that in mind, the game begins, skipping everything leading up to Harry’s arrival at Hogwarts and summing it up in a “story book” mode where you’re presented with dialogue and illustrations detailing it all. The game sets you up with a false sense of freedom from the very beginning, and it won’t take long for you to realize this. There is a set course for you to follow throughout the game, and if you’re not told what to do next from one of the game’s numerous cut-scenes, a little wandering should set you on the right path. You’ll progress through the game in area-specific segments, starting out in the Main Castle, then moving to the Grounds, and then down into the Dungeons before continuing to the later parts of the game. You can talk to your pals Fred and George Weasley and collect Bertie Bott’s Every Flavor Beans for them for a reward, or you can search out the Famous Wizards and Witches cards hidden throughout the castle and the grounds. Collecting the cards is the only real side quest in the game, and you will have to search every new area thoroughly if you hope to find them all. It’s not as hard as you may be thinking, though, and because the game is so linear you won’t find many complex paths to follow and there isn’t any backtracking in the game at all. As for the Every Flavor beans, these can be found all over the place, so as long as you pick up the ones you see, finding the required amount isn’t hard. House points can be earned and lost, but you’ll automatically win the house cup so don’t worry if you’re late for class and you lose a few.
Perhaps the most anticipated aspect of this game was the prospect of being able to use some of the magic seen so frequently throughout the book. In the game, there are a total of 6 spells to choose from, and they can only be used once you’ve learned them. You’ll start with the most basic of spells: Flippendo. You’ll have to attend several different classes throughout the game, only visiting each of them once, which may be a bit of a disappointment to you. Visiting each class a second time would have added a lot more length to the game, but in turn even more would have to be added to the game in the form of story additions and whatnot. Not that it would have been bad, but there’s no need for the game to drag on to that extent.
To learn a new spell, the X, square, circle, and triangle buttons will flash onto the screen and a spark will move from button to button and you have to press the button the spark touches in unison with it. This a lot easier than it sounds because the time window is very lenient. A novelty at first, it takes away all the challenge that learning a spell should have had. It’s very repetitious and not exactly the deepest way to learn a new spell. After you repeat the pattern 3 times, you’re led to a room where you have to practice the spell you’ve just “learned” before you can leave class. It’s always something very simple, and you never fell like you’ve accomplished something after one of these classes. Once you’ve learned one of these spells, using them is even simpler. When a spell can be used against an enemy or an object, there will be a spark floating above it. The color of the spark dictates what kind of spell you have to use. For example, a green spark indicates you’ll need to use the Incendio spell. Press X to activate it, and then depending on which spell you’re using you’ll have to perform some sort of button pressing activity unique to each spell. And then, instant wizardry! Magic is used fairly frequently throughout the game, but some spells will only be used once or twice while others are used every few minutes. Several of the spells feel like they were added to game as a type of filler, only there so you think you’ve learned quite a few useful magic tricks while at the school.
As for the rest of the game, two words: straight platformer. Lots of jumping from here to there, and like Zelda there in no button set aside for this skill. Just run up to an edge and you’ll jump. Pretty far, actually. There aren’t very many challenging gaps throughout the game, and you can use the R2 and L2 buttons to line up the camera for each jump. Besides spell casting, there really isn’t much to do besides run around. Don’t come into the game looking for a challenge because besides Gringotts you could probably blow through this game with ease. After the trapdoor near the end of the game it gets a little harder, but you definitely won’t go anywhere near breaking a controller over the game. There are a few puzzles in the game, but they’re all very laughable. You’re little brother or sister may have a little trouble with the game, but any experienced gamer should fly through it. Playing through the first time, it may take you around 5 or 6 hours to complete, meaning it’s also very short. You can save anywhere you see a red book floating in the air, and they’re placed abundantly throughout the game. It may be a little hard to control with the Analog sticks, but maybe it’s just me. One thing I liked was that you can look around and aim your Flippendo spells through a first-person perspective by pressing Triangle. A great feature that I’m glad they added, and it’s even required in a certain spot in the game. Overall though, the game does nothing to innovate, but even as generic as it is you can’t find complete fault in it. Everything is very basic, unfortunately for you I, but it’s very playable to anyone who comes into the game with an open mind.
With this game, EA had an excellent chance to take everyone’s favorite wizarding sport and turn it into something good enough to be associated with the book or movie versions. In short, it’s no where near as good as we could have hoped for. In fact, it’s barely passable at all and probably won’t hold your interest for any longer than the first few matches. Basically, they took all the excitement and chaos a real Quidditch match possesses and made it as linear and boring as possible. You fly through hoops, and then you catch the snitch. Wow. Great. Excellent. Catching the snitch is also mind numbingly simple. So simple you could very literally catch it with your eyes closed. An icon of the snitch moves back and forth across a bar at the bottom of the screen, and you have to press X at the right moment to catch it. No diving, no fighting off the other seeker, no daring leaps or outstanding catches. Just an incredibly weak attempt to recreate a game that could have been so good. EA, stick to football.
As with the rest of the game, the graphics can be best be described as average. Mediocre compared to Playstation games past and present. The character models are horribly simple. Actually, just plain horrible would work. Very primitive, to say the least. The faces are just as bad, and they only barely resemble their silver screen counterparts. The magic in the game is decent, if not below average, and nothing compared to something you might find in a Final Fantasy game. Animations are acceptable; there’s not much in them to find fault with. Framerate is slow, and sputters pretty often even when you’re the only one on the screen. Textures in regard to the buildings, characters, and outside environments are abysmal. Inside the castle everything is without any sort of shine or flair, and in the grounds and the Forbidden Forest it’s simply terrible. When you’re walking around the grounds, it’s like you’re in a big hallway with jungle colored walls. I feel stupid for even calling it a forest. Jagies are minimal, thankfully, and the castle is of a decent size. A lot to explore, so you shouldn’t ever feel too cramped. The game’s graphics feel and look very rushed and shabby. Not something you would normally find in an EA game. Disappointing to say the least.
Are you starting to see a trend throughout this review and the game? Yes, like nearly everything else in this game, don’t expect much of anything out of the sound department. Music? It’s there, just a little hard to hear. What you can pick up is just your average “mystic quest” theme, with a very soft melody. But most of the time, you’d be better off with a pair of headphones because it’s pretty quiet for a great part of your game. Voices? Yes. Good? Maybe. Do they sound like the actors? No, not really. They’re all imitation, and do sound a little like the real characters if you listen hard enough and really want it to happen. Sound effects? Well, I guess. Maybe all of about 10 sound effects are found throughout the game. Weak is one way to describe them. Barely above SNES quality. Sadly, I’m not joking.
Ok, so I’ve bashed the game for the most part throughout the review. Maybe I have, but what was I supposed to do? Give it a 9 just because it’s Harry Potter? Nah, that wouldn’t be fair to anyone. It gets a 6 only because it’s so mediocre, and not horrible. Little kids will love it, older gamers will shun it. There really isn’t too much about the game to completely tear apart, but it is a very disappointing effort from EA. This game needed a LOT more work, and EA should have considered the older fans of the series instead of throwing something together in a rush to sell a million copies because they knew the little ones would find it very hard to find anything wrong with the game. Yes, Mediocrity is the prevalent theme, and it’ll easily top the charts just because of the name on it. Heck, it could have been a racing game and it would have still sold a million. They took it for granted this time, and if the next game is anything like this I may just decide to pass it up. No matter how good something is, whether it be a movie or a book, you’ll almost always find a game version to be below expectations. With this game, I’d only suggest a purchase for the loyal fan. Not a fan? Only if you’ve got the extra money and you really want to get into the series. Everyone else, find a better game to occupy your time.
Rating: 3.0 - Fair
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No question of cancelling written test for NTs: JKSSB
Srinagar, May 03: The State Services Selection Board has no plans to cancel the written test conducted for the selection of 125 posts of Naib Tehsildars recently and the process for declaration of the results of the written test is on.
Secretary J&K Services Selection Board Ghulam Hassan Sheikh said “a youth who had put a video on social media was approached police and his claim was properly investigated by district police Baramulla. The contents of the video were found totally baseless and just a figment of imagination. The process is on and the results of the written test will be declared at the appropriate time. There is no question of cancelling the written test”.
It is in place to mention here that 80 thousand candidates across Jammu and Kashmir appeared in the written test under tight security and strict vigil at selected examination centres recently but after the test a video that showed leakage of paper. A boy from Shrakwara Baramulla who had uploaded the video was approached by police and after proper investigation the video was found fake and the boy who had uploaded the video admitted that he had done mischief to attract public attention. (KNS)