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On May 7, a shooting incident at the STEM School Highlands Ranch in Colorado caused the death of one student and injuries to eight others. In a recent preliminary hearing, the judge ruled that the prosecution had sufficient evidence to proceed with taking the case against the 19-year-old alleged participant to trial. Reportedly, the defendant claims to have been coerced into committing violent crimes by his 15-year-old friend, who was alleged to have pulled the trigger.
Pueblo County deputies arrested a Colorado man for arson and attempted murder. These violent crimes charges concern an incident in which the 44-year-old man is accused of starting a house fire that could have caused the deaths of the occupants. Reportedly, the deputies were called to the scene of a burning house in the early morning hours of a recent Sunday.
Colorado Springs police responded to a report of armed robbery in the early morning hours of a recent Tuesday. Two individuals were later arrested for suspected violent crimes. Harming someone or attempting, threatening or conspiring to harm someone could be seen as violent crimes, which typically involve robbery, rape or homicide. These acts often include the use of handguns or any other firearms or dangerous weapons.
A 72-year-old Colorado Springs resident was recently arrested for a murder that took place 45 years ago. Advances in DNA technology have led to skilled criminal law attorneys being called upon more and more to defend individuals who are arrested for violent crimes in cold cases. This follows another recent arrest of the alleged Golden State Killer for a murder that was committed 26 years ago.
Anyone in Colorado who faces criminal charges will know that a conviction could mean time in prison. If it involves violent crimes, the consequences can be life changing, including a felony record that will follow a convict forever. Many people might not realize that causing a fatal crash while under the influence of alcohol constitutes a violent crime, even if there was no intention to kill anybody.
At about 3 a.m. on May 18, two juveniles allegedly entered a donut shop in Colorado Springs. They would ultimately be accused of committing violent crimes. Employees of the store claim the two teenagers ordered them to put their hands up, get down on the ground and hand over the cash. They allege that one of the pair became impatient when the cash register was not opened quick enough, and he purportedly struck the employee over the head with his firearm. The gun went off, but no one was hit.
When anybody in Colorado is accused of a crime that involves a weapon, it is typically extremely serious. The reason for this is that the law generally presumes that an individual who possesses a weapon had the intention to use it for other violent crimes. Even attempts to create fear of serious injury in another person by pointing a firearm can lead to severe punishment.
Convictions on criminal charges can lead to many years of incarceration, along with a criminal record and other consequences that can have a severely negative impact on the individuals. However, the laws in Colorado adhere to the presumption of innocence, and the burden is on the prosecution to prove guilt beyond a reasonable doubt. This will also be the case for a recently arrested man who is accused of violent crimes.
Colorado residents in the Pilgrim Downs community were ordered by deputies to stay indoors on a recent Wednesday evening. This followed the discovery of a dead body while law enforcement responded to a call about a suspected home invasion. An incident report indicates that police received a call about a possible break-in at approximately 10:30 p.m. Upon checking the residence for burglars, deputies came across the victim's lifeless body, and the possibility of further violent crimes caused the call for the residents to remain inside their homes.
For a person to be convicted of a criminal charge in Colorado, the prosecution must prove guilt beyond a reasonable doubt. The defendant has the right to present a defense during the trial to raise a reasonable doubt. This will be a challenging process, particularly in cases involving violent crimes.


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A Harmful Way to Eradicate Pests
In our everyday lives, there are many products that have been introduced over time that are designed to solve problems, but in doing so, they can actually be quite harmful for human consumption or ingestion. A prime example of this is of course, pesticides.
Pesticides have been designed to eradicate pests that can cause damage to crops or forests, cause damage and spread disease in playgrounds and parks, within our homes and communities, and some that can even kill us. But one thing about pesticides, is that they almost always have risks associated with them, no matter what their purpose. Pesticides include chemicals that control weeds (herbicides), kill insects (insecticides), eradicate fungus (fungicides), rid your property of rodents (rodenticides), and others.
Despite the importance pesticides play in a consistently balanced existence here on planet earth, science is constantly uncovering new information relating to pesticides, and the health implications connected to exposure to pesticides that often causes alarm, and with good reason. The harsh reality about pesticides is, that they do more damage than good.
Even though we learn new things about these poisonous chemicals we put on our food and in our water sources, it still does not stop us from using 2.5 billion kilograms (5.6 billion pounds) of pesticides globally every year. With more than 17,000 pesticide products on the market, the fact that they are such big business means they will probably never go away.
Connections To Disease
The connection between common diseases and pesticide exposure continues to strengthen despite efforts to limit exposure, or weaken chemical risks. Common diseases like birth defects and reproductive dysfunction, asthma, autism and learning disabilities, Parkinson’s, diabetes, and Alzheimer’s and many types of cancer are all known to have connections to pesticide exposure. If that is not enough reason to avoid exposure, then we need to go a little further in depth.
Alzheimer’s disease affects around 10% of the population over 65 years old, and evidence is constantly growing that pesticide exposure has a role in its development. Some pesticides increase beta-amyloid peptide which are key characteristics of the disease.
As the number of people suffering from asthma continually increases, so too does research linking pesticides with asthma. There are hundreds of studies available that show links between chemical exposure and asthma, bronchitis and other respiratory problems.
Numerous pesticides have been known to have a negative impact on fertility in both men and women. Chemicals like methoxychlor, which is used on food crops as well as home gardens lowers women’s chances of becoming pregnant with exposure. Low sperm counts and altered hormone levels are linked to infertility in males with exposure to organophosphate pesticides.
Other life-threatening diseases that are linked to chemicals commonly found in pesticides are non-Hodgkin lymphoma, occupational skin disease, and Parkinson’s disease. Parkinson’s disease has had many linking studies to chemicals like the pesticides paraquat and rotenone.
It’s Not Just Adults or Humans At Risk
Not only are humans at risk of harm when exposed to chemicals found in pesticides, but our pets are also at risk of harm. Household, lawn, and garden pesticides can be dangerous or deadly to your pets, including cats, birds, dogs, reptiles, horses, and fish.
Pesticides aimed at cats and dogs, like flea and tick products, can be very hazardous to your pets, even when used as instructed. It’s very important to know that the chemicals used to keep fleas off dogs, is toxic for cats, so you need to ensure you don’t mix your pet’s flea control pesticides. Your pets are also at risk when you control or treat areas of your yard for pests like weeds, and can end up dead themselves if they happen to happen to consume a mouse or rat that has been poisoned.
Children are particularly vulnerable to the risks of pesticide use. Because the human brain is not completely formed until around 12 years old, childhood exposure to pesticides can impact the development of the central nervous system. Because children have not developed their immune systems, detoxifying mechanisms or nervous systems completely, simple things like playing in the grass, putting things into their mouth and even playing on floors can increase the effects of pesticide exposure.
One of the most important reasons we should actually avoid pesticides (and their use) is that they often kill off beneficial species as well as the pests they were intended for. Two of the most beneficial species on the planet are heavily affected by pesticide use, and those are bees and worms.
Bees of course, naturally pollinate plants. Without them, we don’t have plants. Not only do they pollinate natural plants, they pollinate farm crops, and forage plants which livestock rely on to survive. Bees are quite possibly the most important creature on the planet, and without them, we will have no food crops, and our oxygen will run out. Pesticides that are harmful to bees are in fact putting the lives of every human on earth at risk.
Worms maintain soil health and reduce the number of pests naturally, and without them, farming would be almost pointless. Worms help to maximise the amount of water and that filters into soil. They break down organic matter into natural fertilizers. Pesticides in the soil often kill off worms, making the farming job harder than it should be.
Despite what pesticides have been generated for, if we don’t avoid them, our future will be impacted. No matter what continent or country you live in, pesticides are readily available, and only you can make the choices to avoid them as much as you possibly can.

domain: agronomy


Trampoline Parks Are Dangerous. Last night, Steve Stoler did a story for WFAA about how dangerous and unregulated trampoline parks are. Kind of a “no duh.” My 14-year-old is still recovering from a broken ankle sustained at Jump Street. After the City Council shuts down Uber in Dallas, I suggest they turn their attention to trampolines.
Man Drowns in Golf Course Pond. A maintenance worker was trapped under a mower and drowned in a pond at The Courses at Watters Creek, in Plano. This was the second drowning at the golf course this summer. It’s obvious what needs to be done: the City Council needs to rewrite Dallas’ code to outlaw golf course ponds. I know the Dallas City Council has no authority over Plano. But it should.
Face-planting Skateboarder Is at Large. Flower Mound police are looking for a skateboarder who tried to jump a gate at a city park but didn’t make it and instead broke the gate on his way to landing on his face. You can watch video of the skateboarder failing. Note to Dallas City Hall: skateboards and gates ought to be outlawed.


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To crack Adobe Photoshop, you first need to locate a program that is called a keygen. Keygens are software that will generate a valid serial number for the software. You can download one from GetMxy.com . After downloading the keygen, run it and generate a valid serial number for Adobe Photoshop. Once the serial number is generated, you can then launch Adobe Photoshop and enter the serial number to unlock the full version of the software. Adobe Photoshop is now fully functional.
To crack Adobe Photoshop, use the steps in the tutorial above. First, download Adobe Photoshop Serial Key Generator . This is a software that will generate a serial number for you. Next, locate the patch file and copy it to your computer. After this, you need to open Adobe Photoshop. You can then enter the serial number to unlock the full version of the software. Finally, you can start using the software to do whatever you want.
A small transition of the arrow from a vertical to a horizontal line marks when you click “Reset” at the bottom of the dialog. The layer controls have been replaced by a single button in the upper-left of the screen (“Create New Layer”). You can collapse the panels into the “Arrange” or “Zoom” panels, which are at the top and bottom of the dialog, respectively.
Click the “High Resolution.eps” link on the right to view or download the mockups at larger sizes. You can even use them as backgrounds for your website, or for other purposes where high-quality images are required.
Our best selling mockup is a combination of Leather and High-Resolution EPS files. All elements are super easy to insert right into your final website. With multiple color schemes and detailed designs these mockups have this sneaker stash website covered. Sketch your website.
All of these changes happen along the path of history. Adobe was one of the first photo editing makers, but it’s now largely just a commodity. Photoshop has been around that long, and it shows. I’ll be updating this photo review once it’s finished, so keep checking back.
Whether you need a standalone photo editor or one that works with Photoshop, Lightroom, or Photoshop Touch, Adobe Photoshop Elements 2018 is the program for you. It’s a great balance between power and simplicity. The program is equipped with everything you need to do any kind of photo editing, including RAW conversion. The result is that with Elements, you have no excuse for not getting a good photo.
The number of different photos and image editing software programs is a lot. This includes both Windows programs and the Mac operating system as well. Adobe Photoshop and other software programs are the most used programs in image editing.
If you know what you’re doing, editing and retouching photos with Photoshop can be rewarding and fun. However, if you’re new to the process, you may feel overwhelmed by the endless amount of options and preferences that are available in the program. With that in mind, we’ve created this step-by-step guide to help you use Photoshop productively.
A powerful adobe photoshop desktop app was available. And we’re excited to announce that in addition, Photoshop for Mac is now free for one year. We’re bringing new features to Photoshop, including an updated interface, a powerful content-aware fill tool, and a powerful new tool to help us better understand the content of an image, Photoshop GPU technology, and support for macOS Mojave. The free version of Photoshop for Mac will be available as an update to everyone.
With every new technology we witness an endless stream of new applications, some of which prove to be revolutionary and useful. Today I’m going to speak about Adobe Photoshop CC as an image editing software for illustrators and designers.
What It Does: The Dodge & Burn filter technique is used to make lighter colors darker, or dark colors lighter, producing a more multifaceted, depth-filled image. With the Smudge tool, you can blend the color of one area with that of the surrounding area, producing a new color gradient. Photoshop is one tool in a large, complex collection of editing equipment, and sometimes people don’t even realize how important they are to what they do. For example, the lighting adjustments and the HDR adjustments (different things in post-processing programs).
Adobe Photoshop has a new version on this year, after a very long wait. The new version of this software is called Photoshop CC and its official release is on 30 june,2018. Because this is a complete course it is very important to learn and adopt the new features of this software in order to save your time and efforts to learn and apply.
Photoshop is a standard tool that allows editing and manipulation of images and pixel-based graphics elements. Adobe Photoshop is a vector graphic program usually associated with the creation of visual display effects. While Photoshop is certainly that, it can also be used to enhance images, expose image for quality and correct color or exposure. Photoshop has a wide range of area in which photographs are processed, from transforming an image from its original to that of an abstract, or adding a texture or distant. It is considered a standard workflow for graphic and illustration designers, including photographers wanting to manipulate a photograph in digital.
Photoshop features some of the tools to create a picture without any use of models and photographs, such as sketching and painting. It has a set of tools and collections of effects for the image editing. They are canvas and area tools, which can be applied to other features. For instance, you can draw without the need to use a pencil and paper, or paint a cover. The pixel is a measurement of any display, computer, or print format. The pixel is divided very small areas; usually, the pixel is one dot. Photoshop is not just a drawing tool, but it also has areas effects such as the color effects, pattern effects, gradient and brush effects, strokes, blend modes, smart objects, filters, retouching and more tools for a more productive process.
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Other new features in Photoshop for 2021 include the Preview Skill as a Photoshop default, which layers art you’ve previously created as preview layers in the background of your photo. Paint Layer Features returns, giving you the ability to adjust settings directly in Photoshop, without having to switch back and forth between the layers panel. This is great if you want to add filters or adjustment layers to an image, but don’t want to leave Photoshop. You can toggle the Preview Skill for the current editing session with the eye symbol on the top left, and the option to switch to the Preview Skill when no sessions are open (it seems like this feature will be fully supported in Eyes, presumably an upcoming version of the Photoshop for Laptops app ).
The addition of the ability to create and save Creative Cloud Libraries (CCLs) gives you the ability to share your entire library of custom brushes, patterns and textures with other users. CCLs can be 200MB, 1GB or 10GB in size. Additionally, Photoshop Elements 2021 adds support for online and offline usage. You can use Photoshop Elements online only and don’t need to ever download your entire library of customized items – a significant increase for the convenience of having all your brushes and patterns accessible, regardless of whether you are logged on or offline in the app.
The most useful features from Camera Raw in versions 9.0 and above are also heading to Photoshop. The most significant of those is the new Adjustment Brush, which you can use to directly adjust layers with tools such as Inner Shadow or Clipping – and even mask areas to dramatically change them.
Adobe Premier Pro is an advanced feature-packed video editing application for integrated graphics and media creation. It includes the best integrated media cameras and innovative tools that enable you to complete your work in a streamlined professional way.
The Adobe Experience Manager (AEM) workflow platform is the leading enterprise content management (ECM) solution that ensures content is securely, reliably, and seamlessly managed for people, processes, and business initiatives. AEM framework technology is extensible, making it easy and cost-effective to create websites, online stores, social network sites, mobile, and other content-rich applications.
It’s hard to throw away Photoshop Elements. Still many of the best features of Photoshop, like local adjustments, layers, the ability to customize Toolbars and Guides, and even a few adjustment tools are favorites of Photoshop Elements. To enable easier access to your favorite Photoshop features and find shortcuts to get things done faster, you’ll find a few shortcut features available in Photoshop Elements 2015 that aren’t always apparent. Once you’ve started using Photoshop Elements, you’ll find many new features including new brushes, frames, and tools, together with a new dark user interface. Photoshop Elements is great for beginners and even some advanced users, so don’t throw out the old and in.
While most people don’t think about using light boxes as a way to display artwork, they’re a great way to ensure a professional, studio-like feel. Photoshop Lightroom 4 includes a new Lightbox feature that lets artists use a light box to display artwork for clients, as well as a unique online view. You can even take images directly from Lightroom and use it for a show. You can also give feedback on your website, and you can choose from the Lightroom community and online plan to have Lightroom create a job order for you.
Adobe Photoshop is a powerful graphics editing software, used to create digital images from scratch. A photograph is not just a flat drawing; the photograph is actually a series of tiny dots that define the color, tone and brightness of the image. Adobe Photoshop is used to define and create these tiny dots. Most of the images on the web are created using Adobe Photoshop. Just like most Photoshop features all the editions of Photoshop can be used to create customised, unique and remarkable images.
Photoshop is only one product among many offered by Adobe. This is the flagship of their graphic design products and the definitive solution to any creative imaging challenges. Neatly enough, it is a recursive acronym for “Photoshop Extended”. It is very famous for its many attractive features that will help users in editing and correcting images.
As a matter of fact, Photoshop is the best designer’s tool, which is used to modify images in various ways. The user interface, which is easy to handle, is an essential part of Photoshop. This digital tool is perfect for simple images that need correcting but still sophisticated for really tough tasks.
There are many different types of photo editing tools such as photo cropping, photo enhancing, and photo retouching. One of the main functions of these tools is where you can edit and add effects to images.
After the introduction of the free version of the software called Photoshuper.net, it gained a lot of popularity among software users. The free version of Adobe Photoshop is easy to learn and simple to use. It has improved many of Photoshop’s features, making them much easier to use. The software can also be used as a creative tool which can be used by all types of professionals.
Praise for the program has long been fairly spotty. A recent study by Gartner ranked Adobe as the undisputed world leader for the fourth year running. Microsoft’s “The only program you can’t do without,” however, is far more positive. “I dare you to find a professional photographer or other graphics specialist who thinks any other piece of software delivers more versatility and power than Photoshop CS6”, on the company’s website.
Although it has been designed for personal use, this version is aimed at serious gaming enthusiasts. In the vein of the work of the Game Theory Explained series, this book analyzes using these games to learn about game theory, strategic and fairness games, and better ways of playing games.
Game theory provides the foundation for games, to make sure they are trustworthy and fair. With that knowledge, people can play games without fear they will be cheated or defrauded. We can even learn to win more effective strategies!
The Game Theorists books are aimed at the readers who wish to learn about games in a fun, simple way. They probe the inner workings, hidden motivations, and games at a deep strategic level to help the reader achieve victory.
With every new version, Adobe may introduce some new feature in its product, but there are some tools and features which make a great difference for users. Here are some tools and features which are proved as the best of Photoshop:
The most important tool to any designer is the original brush tool. In Photoshop version 13, Adobe added multiple brush modes like Blob Brush or Liquify. You can now change the brush size in any direction, and the brush mode will be accurate in terms of its size. No matter what you have had in old Photoshop, it is now easy for you to enhance your image in Adobe Photoshop.
The Photoshop CC 2019 for Desktop and Extended Features is the perfect program for managing large or medium-sized operations. A user can easily navigate around Photoshop CC 2019. The first impression of the program is its well-rounded interface and its speedy performance. To find more options such as in the panel options, click on the window that you would like to edit. To get more tab features, select the workspace, and choose the desired one. It will automatically open the program in a tab. It’s extremely convenient to work on the program. You can also edit files with the help of a requirement to reduce the file size.
The new features of Photoshop CC 2019 is something worth to know. You can get all the new features in one graphic tool. The new release of Photoshop comes with two new features, called the Steam location and Siri. The use of the Steam location is a new feature in Photoshop CC 2019. It is a space on your desktop or laptop screen where your most-recently-opened files will appear. You even can access this document folder through Photoshop. There are several features that you can do on the Steam location folder. A user can share this folder on social media or even the web. It will be a wonderful addition to all of the PSD files in the folder.
The power of the tool is quite fascinating and consequential. It is a graphic editing software which makes editing of images done in a very easy and efficient way. Furthermore, it can be used to create different kinds of images like paintings, photography and videos.
Adobe Photoshop Portable /Photoshop Express: In the early days, Adobe Photoshop’s vision was to open the doors to creative leadership for the next generation of photographers. And to help do that, the company produced Photoshop Portable, its first portable version of Photoshop.
If you struggle with the concept of “Photoshop” as an actual program (akin to something like “Word”), then I have good news for you. Photoshop Elements is the solution offered by Adobe, and while you will still use the program for its main features, so will I. In fact, you’ll find me using it just like you use any normal photo editing software.
If you have struggled with basic photo editing for years, then Photoshop Elements should be right up your alley. It makes a lot of the fun Photoshop effects more accessible to the nonprofessional, but you’ll still need to do some tweaking to get the best results. As with all the Creative Cloud applications, Photoshop Elements 2019 is a member of the Creative Cloud family, and you’ll be able to make the most of your new subscription by adding Subscription Services and the App Gallery. And for that price, it’s an absolute steal.
If you learn this straight from the one of Photoshop’s most respected authors, you can learn the basics of Photoshop in just a few short hours—and maybe even get a little more done than you would in similar spans of time working with other software. It’s worth it. K.S. Wolff takes a painstakingly clear-eyed approach to this second edition of the bestselling book—and while there may be some new things in here, they’re not exactly earthshaking. But if you’re looking to turn your doodles into a paper cut-out or a digital painting, this is the book for you.
Blend Modes – These were introduced with the version CS6 with the open filter name. This tool easily creates a wonderful interplay of sorts between your images. The blend mode is known to create one of the best tools that are all about changing colors. Not only that, it helps you in creating different styles. It is possible to make your photos more crisp, bright, and vibrant with the help of this tool.
Camera Raw – This was bad the previous version but is now better.
Color Correction – This tool changes colors and tones to the one of your choice. Not only that, you can change the brightness, contrast, exposure, and saturation. It can be a blurry face or an over-exposed photo. It can use the healing brush, clone stamp, or adjustment layers to bring out the correct details of your work.
Sometimes, the second version of any product is so much better than the first that it’s almost revolutionary. As a result, Adobe Photoshop CS2:
- Went from being an expensive and not a popular tool to being a popular and affordable tool for all types of photographers
- Went from being a complex and tough user interface tool to being really easy to use, even for beginners
Adobe has been continually pushing the envelope in image editing software, and its developers clearly know where and how to make improvements. As was the case with Photoshop, the transition to a tablet-based software was accompanied by a shift from using a mouse to a stylus.

domain: design


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This file is not currently available to purchase on Tradebit
Sold by music on Tradebit The world's largest download marketplace 2,956,626 satisfied buyers
MP3 hi-five - ROCK: 60's Rock
rock and roll with a gulf coast twang
11 MP3 Songs
ROCK: 60''s Rock, ROCK: Surf Rock
Listening to this record, you can tell how much we wanted to capture the sound of our live shows. We were playing really well on a Midwest tour and felt up to the challenge of recording while on the road. We had a gig in Chicago one night and decided to record our set a few times before the show. The end result is a gritty rock and roll record made the old-fashioned way.
"The hi-five album draws on many familiar styles of classic rock. It is something of what the Shins did with Brian Wilson, but with the Beatles. The album ranges from an acoustic neo-folk in Jack Johnson''s style, with "family name" and "bros. kennedy," which glorify tradition, to more upbeat rock, with wandering guitar solos and amp effects similar to the Pixies"
The Review, Houston TX
"I just did a re-listen of hi-five. These are some of the great songs of our times. "border lights," "stranger," and all the others...What I am
getting from these songs is positive vibrations . . . I get a sense of
"joie de vivre" from your music. "Elan vital," if you will. "
This file is sold by music, an independent seller on Tradebit.

domain: artistic


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|Sizemore to Co-Chair Committee|
FOR IMMEDIATE RELEASE
CONTACT: Ruwena Healy
Marketing 24/7, Inc.
SIZEMORE TO CO-CHAIR COMMITTEE FOR
AMERICAN BAR ASSOCIATION
BIRMINGHAM, Ala. – July 21, 2011 – Alton Sizemore, a forensic accountant and Director of Investigations with Forensic/Strategic Solutions, PC, has been named as co-chair of the American Bar Association’s Southeast Regional Litigation Support Committee within its Criminal Justice Section. Sizemore will serve as co-chair with Keith Keller of Duff & Phelps. Together, they will serve the ABA during its fiscal year from July 2011 to June 2012.
The Southeast Regional Litigation Support Committee falls under the Associates Committee of the Criminal Justice Section of the ABA. Its goals are to increase associate membership, increase activities designed to achieve that goal by way of conferences and other educational opportunities, and serve the interests of the associate members in the litigation support field.
“Being involved in this committee deepens my understanding of the legal profession's needs," Sizemore said. "It also helps lawyers and those in litigation support roles better understand each other's services. This happens through attending conferences and other means of education.”
About Alton Sizemore, Jr., CPA, CFE, CFF
Alton Sizemore, Jr.specializes in the areas of fraud examination and investigative financial consulting with a focus on the banking industry. He works closely with plaintiff and defense attorneys, governmental agencies, government inspector generals, and corporate boards,.
Sizemore joined Forensic/Strategic Solutions after retiring from the FBI with 25 years of service. Sizemore served as Assistant Special Agent in Charge of the Birmingham division of the FBI, managing programs for Counterterrorism, Foreign Counterintelligence, Cyber Crime, Public Corruption, Civil Rights, White Collar Crime, Violent Crimes/Major Offenders, and Drug Fraud. His broad range of experience with the FBI includes criminal investigations in white-collar crime in many areas, including business, government, medical, and banking.
Sizemore is a member of numerous professional organizations including the Metropolitan Criminal Justice Executive’s Association (MCJEA), the American Institute of Certified Public Accountants (AICPA), the Alabama Society of CPA’s, and the Association of Certified Fraud Examiners (ACFE).
He is a sought after speaker as well as an instructor for the Association of Certified Fraud Examiners and the American Institute of CPAs. He presents programs on numerous topics, including identifying fraudulent financial transactions, investigating workplace embezzlement and what to expect when investigated by the FBI.
About Forensic/Strategic Solutions, PC
Forensic/Strategic Solutions, P.C. is a national financial investigation firm that combines fraud examination with investigative financial consulting services in order to illustrate and present complex financial data in courts of law and other forums. Our deep expertise in data analytic techniques and litigation consulting results in fact-based, clearly-illustrated data, best positioning our clients to achieve favorable outcomes. Forensic/Strategic Solutions was founded in Birmingham, Alabama in 1992.

domain: finance


Lexus Working On EVs With A Full-Manual Transmission | V3Cars
At the Kenshiki Forum 2022, Lexus announced that they are working on an electric SUV with software-based manual transmission. However, unlike an automatic transmission with manual control through shifter lever or paddle shifters, the Lexus EV will offer a full manual transmission driving experience. It’ll have a proper H-pattern shifter along with clutch pedal, a tachometer and sounds in the cabin to simulate driving a manual transmission.
The only caveat here is that it’s entirely software based. There’s no mechanical linkage between the shifter lever and the drivetrain although there are gates to give the feeling of slotting the shifter in the desired gear. Lexus chief engineer claims that the torque from the motor changes based on the gear selection. This way, the experience is similar to driving a manual transmission. Lexus even say that you can access different maps to get a completely different driving experience. It’s even possible to simulate the torque delivery of completely different cars and get a different driving experience in a single car.
Note: Check your Car EMI with our - Car Loan EMI Calculator
The chief engineer of Lexus, Takashi Watanabe, said that the manual electric vehicle is beyond the concept phase and Lexus are seriously considering to start delivering it in a production vehicle. Currently, they’re testing it on the UX 300e electric SUV.
Watanabe added, “From the outside, this vehicle is as quiet as any other BEV. But the driver is able to experience all the sensations of a manual transmission vehicle. It is a software-based system, so it can be programmed to reproduce the driving experience of different vehicle types, letting the driver choose their preferred mapping.”
Besides a manual transmission, Lexus are also testing software-based steer-by-wire and brake-by-wire systems for mass production. Eventually, we can expect these features to trickle down to the more affordable Toyota vehicles.
Currently, no electric vehicle comes with a manual transmission from the factory. Only the Porsche Taycan and Audi E-Tron come with 2-speed automatic transmission to maximise efficiency and high-speed performance.
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It doesn’t matter if you have been in your current position for several years or a few months. We all get the same dreaded feeling when we have to ask someone for help.
We begin doing the tear ourselves down thing. I should know this. Why can’t I figure this out? I am so stupid to think I could do this job. If I ask for help, I look dumb. My co-workers and boss will really think I can’t do my job. It is awful how we do this to ourselves.
In our mind, we don’t want to look stupid, so we don’t ask for help. We go ahead and complete the entire project to the best of our ability and it is all wrong. It is going to be presented to a client……wait a minute, let’s rethink this plan.
So you have to complete the project that is going to be presented to a client. Do you really want to be the one responsible for a messed up project? I don’t think so. It’s difficult to admit when you don’t know something. We think we should know how to do everything in the scope of the position we fill.
We need to accept and understand that it is okay to ask for help. It doesn’t make you appear or look dumb. Being able to admit that you need help is the first step, and the next step is asking for help. People have much more respect for you knowing that you are willing to ask for help. Many times you can just ask a co-worker to help you and they will. This will also make them feel comfortable asking you for help in the future should they need it.
It is important that you feel comfortable enough that you can go to your boss and ask for help. It is part of the boss’s responsibility to assist you when you need help with something. They are there to provide training when needed. Remember, they are the boss, managing and teaching is part their job. If they are a good boss, they will encourage you to be the best that you can be and assist you in whatever way they can so you can meet your goals. A good boss empowers the employees to be able to work independently and be able to work in a team setting, too. You can always ask your boss for help. Don’t be afraid to ask. Trust me, they will be angry if you needed help and didn’t ask them. I’m sure the boss has had times when he or she needed help, too. No one person knows “everything”.
Just remember, don’t be afraid to ask for help. It’s part of the learning process. We all have needed and asked for help at some point in our career.
So just make it easy on yourself and get over it. If you need help just ask!
Your comments are encouraged and appreciated.
Image courtesy of Danilo Rizzuti, thank you.

domain: sociology


text:
Microglial motility in Alzheimer’s disease and after Aβ42 immunotherapy: a human post-mortem study
Acta Neuropathologica Communications volume 7, Article number: 174 (2019)
Microglial function is highly dependent on cell motility, with baseline motility required for homeostatic surveillance activity and directed motility to migrate towards a source of injury. Experimental evidence suggests impaired microglial motility in Alzheimer’s disease (AD) and therefore we have investigated whether the expression of proteins associated with motility is altered in AD and affected by the Aβ immunotherapy using post-mortem brain tissue of 32 controls, 44 AD cases, and 16 AD cases from our unique group of patients immunised against Aβ42 (iAD).
Sections of brain were immunolabelled and quantified for (i) the motility-related microglial proteins Iba1, cofilin 1 (CFL1), coronin-1a (CORO1A) and P2RY12, and (ii) pan-Aβ, Aβ42 and phosphorylated tau (ptau). The neuroinflammatory environment was characterised using Meso Scale Discovery multiplex assays. The expression of all four motility-related proteins was unmodified in AD compared with controls, whereas Iba1 and P2RY12, the homeostatic markers, were increased in the iAD group compared with AD. Iba1 and P2RY12 showed significant positive correlations with Aβ in controls but not in the AD or iAD groups. Pro- and anti-inflammatory proteins were increased in AD, whereas immunotherapy appears to result in a slightly less pro-inflammatory environment.
Our findings suggest that as Aβ appears during the ageing process, the homeostatic Iba1 and P2RY12 –positive microglia respond to Aβ, but this response is absent in AD. Aβ-immunisation promoted increased Iba1 and P2RY12 expression, likely reflecting increased baseline microglial motility but without restoring the profile observed in controls.
Evidence from genetics, experimental and human post-mortem studies  supports the involvement of microglia, the resident immune cells of the brain, in the pathological cascade that leads to the development of Alzheimer’s disease (AD). However, their role in the disease is still not fully understood . In the healthy brain, microglia survey the parenchyma, constituting its first line of defence against pathogens or injury [10, 34], and interact with neurons helping to shape synaptic communication [29, 52,53,54, 56].
One of the most prominent characteristics of microglia is their motility, which is essential for the cells to perform their functions. Two types of motility are present in microglial cells: baseline motility, which consists of the extension, retraction, and movement of the microglial processes [18, 37], allowing microglia to survey their environment, clear cellular debris, interact with neurons and synapses, and remodel the extracellular matrix . If, while performing this baseline motility, a microglial cell encounters molecules that could be indicative of infection or tissue damage, the cell will then switch to directed motility. Directed motility consists of a targeted extension of processes towards the source of injury . At the molecular level, microglial motility depends on two mechanisms: i) reorganisation of the actin cytoskeleton and ii) purinergic signalling .
The actin cytoskeleton consists of a dynamic filament network, regulated by a complex machinery that involves several proteins. Some of the most important actin-related proteins in the microglial cytoskeleton are the coronins, the cofilins and the ionised calcium-binding adapter molecule (Iba1) [1, 46]. The coronin gene family contains six members with roles as regulators of actin dependent processes, such as cell motility and vesicle trafficking [40, 41]. Coronins localise to sites of dynamic actin assembly, regulating filament organisation via interactions between actin and the Arp2/3 protein complex. This results in the building of branches in the actin filament network . Coronin 1a (CORO1A) plays a large role in the normal immune system as the most prominent coronin in cells of haematopoietic origin , and has been previously identified in human microglia . Cofilins, and particularly cofilin 1 (CFL1), are actin-binding proteins that depolymerise and separate actin filaments . Lastly, Iba1 is a cross-linking protein, the role of which is to organise actin filaments into networks. In microglia, Iba1 is crucial for the formation of actin bundles, , an event necessary for the construction of lamellipodia, filopodia, and membrane ruffles. These structures are essential for microglial migration and phagocytosis. Due to its relatively homogeneous distribution along the cell body and processes, Iba1 has been widely used as a marker to identify microglia in humans and experimental models. Purinergic signalling plays an important role in microglial motility. The presence of extracellular nucleosides (ATP, ADP, UTP, UDP), released as a result of neuronal death, triggers the activation of P2Y receptors. The ADP receptor P2RY12 is now regarded as a microglia-specific marker, mainly associated with directed motility, driving chemotaxis through a mechanism that involves the activation of potassium (K+) channels . Therefore, both actin reorganisation and purinergic signalling are essential to the two forms of microglial motility.
Alterations of microglial motility have been documented in AD, with (i) microglial clusters identified around Aβ plaques indicating movement of microglia as part of their response to Aβ aggregation [16, 57], (ii) in AD mouse models, impairment of directed motility and phagocytic activity , and (iii) morphological changes in plaque-associated cells . Aβ-immunotherapy is one of the major current therapeutic approaches being investigated in AD . The first clinical trial of Aβ immunotherapy in humans involved active immunisation with a full-length synthetic Aβ42 compound named AN1792 . Our neuropathological study of these patients has shown a markedly reduced plaque load associated with microglial changes [57, 58]. However, these changes were not associated with reduced cognitive decline [17, 33].
In this study, we have explored whether the expression of proteins associated with motility is altered in AD and affected by the Aβ immunotherapy using post-mortem brain tissue of controls, AD cases, and our unique group of AD cases immunised against Aβ42.
Material and methods
Brain tissue from 76 donors was sourced from the South West Dementia Brain Bank comprising 44 AD cases and 32 controls. Alzheimer’s cases had a clinical diagnosis of sporadic AD made during life and satisfied post-mortem neuropathological consensus criteria for AD . Cases with any other significant brain pathologies such as stroke, tumour, or traumatic brain injury were excluded from the study. Controls were aged-matched cases, with no history of neurological or psychiatric disease or symptoms of cognitive impairment. Additionally, 16 immunised AD subjects (iAD) who participated in the AN1792 clinical trial were also included in the study. The characteristics of the groups are presented in Table 1 and individual details in the Additional file 1: Tables S1, S2 and S3.
Cerebral cortex from the inferior parietal lobule was investigated in all cases. This is one of the areas recommended for neuropathological assessment by the CERAD (Consortium to Establish a registry for Alzheimer’s Disease)  as it is significantly affected by AD pathology. Formalin-fixed paraffin embedded tissue was used for the immunodetection of neuropathological and neuroinflammatory markers. Frozen tissue, available for 31 controls, 35 AD cases and 11 iAD cases was used for detection of inflammation-related proteins by Meso Scale Discovery multiplex assays.
Immunohistochemistry was performed on 4 μm paraffin-embedded sections, targeting the markers of AD pathology pan-Aβ (clone 4G8, BioLegend), Aβ42 (clone 21F12, Elan Pharmaceuticals Ltd) and the phosphorylated (p) tau (clone AT8, ThermoScientific) protein, as well as the microglial motility-related proteins Iba1 (rabbit polyclonal, Wako Chemicals), CFL1 (polyclonal rabbit, ThermoScientific), CORO1A (rabbit polyclonal, LifeSpan Biosciences) and P2RY12 (rabbit polyclonal, Sigma Aldrich). Bound antibodies were visualized using the avidin–biotin–peroxidase complex method (Vectastain Elite, Vector Laboratories) with 3,3′-diaminobenzidine as chromogen and 0.05% hydrogen peroxide as substrate (Vector Laboratories). All sections were counterstained with haematoxylin, then dehydrated and mounted in Pertex (Histolab Products AB). The staining was performed in two batches with each batch containing cases from all groups (Control, AD, iAD). All experiments included a negative control slide incubated in buffer with no primary antibody and a positive control slide containing a specific tissue type known to express the protein of interest (e.g. tonsil).
Quantification of staining
Quantification was blinded to the case designation and performed on the grey matter in the same sulcus of the inferior parietal lobule for all cases. For each antibody, the slides were scanned together at a magnification of × 20 in an automated slide scanner microscope Olympus VS110 (Olympus America Inc.) and visualised with Olympus VS-Desktop software using a specialised add-in that allows the extraction of the region of interest (ROI) of 0.25mm2. For each slide, 30 ROIs were selected in a zig-zag pattern within the grey matter, ensuring coverage of all cortical layers. Extracted ROIs were analysed with the ImageJ software (version 1.49u, Wayne Rasband, NIH, USA) using an automated macro specific to each antibody. The area fraction labelled by the antibody in each ROI was obtained by quantifying the presence or absence of the staining in each pixel, expressed as protein load (%).
Inflammatory proteins were measured on the V-Plex Meso Scale Discovery (MSD) electrochemiluminescence multi-spot assay platform (MesoScale Diagnostics, Rockville USA). 100 mg of frozen grey matter were homogenised at a tissue concentration of 20% w/v in RIPA lysis buffer (Thermo Fisher Scientific), supplemented with protease inhibitors (Sigma Aldrich) and phosphatase inhibitors (Thermo Fisher Scientific). Total protein concentration in the supernatant was measured by BCA Protein Assay Kit (Thermo Fisher Scientific). 25 μl of brain homogenate (1:2 dilution) were used for the V-PLEX human proinflammatory panel 1 and cytokine panel 1, and each plate was read on a Meso Quickplex SQ120 (Meso Scale Discovery) according to manufacturer’s instructions. Absolute target protein levels (pg/ml) were obtained and normalised with respect to the total protein concentration, as obtained by the BCA assay. Three frozen homogenates from a multiple sclerosis brain containing either chronic inactive, acute or chronic active lesions were included as positive controls.
Statistical analysis was performed using the IBM SPSS v24 statistical software package (SPSS Inc. Chicago IL). The normality of distribution of each marker across the groups was assessed by the Shapiro-Wilk test. Data were not normally distributed, and thus non-parametric tests were used. Comparisons among study groups were performed using the Kruskal-Wallis test. The Spearman’s test was performed to assess correlations between variables – to explore the relationship (i) between microglial motility proteins and key features of AD pathology (Aβ and tau loads), and (ii) between the microglial-motility proteins. To account for multiple testing, the Benjamini-Hochberg procedure to control for the false discovery rate (FDR) was used as post-hoc correction. For all tests, an adjusted P value < 0.05 was considered significant.
Aβ and tau pathology
Aβ and ptau immunoreactivity was quantified in order to characterise the cases with respect to AD pathology and to permit correlations with the microglial motility-related proteins. A significantly increased pan-Aβ load was observed in AD compared to control cases (P < 0.001), whereas it was significantly decreased in the iAD group compared to AD cases (P < 0.001), as expected. No significant difference was observed between control and iAD cases (Fig. 1a). Aβ42 load was significantly increased in AD vs control cases (P = 0.006) and decreased in iAD vs AD cases (P < 0.001), as expected. Interestingly, Aβ42 load, was also significantly decreased in iAD compared to control cases (P = 0.021) (Fig. 1b). Ptau was significantly increased in AD vs control cases (P < 0.001) and decreased in iAD vs AD cases (P = 0.026), with no difference between the iAD and control groups (Fig. 1c).
Microglial motility-related proteins
Immunohistochemistry for the four proteins related to microglial motility was investigated in the cortical grey matter. Iba1, CFL1 and CORO1A immunolabelled microglia and perivascular macrophages and in AD microglial clusters around amyloid plaques. While P2RY12 was specific to microglia with no labelling of perivascular macrophages (Fig. 2).
Quantification of these four proteins showed no significant differences between AD and the control group: Iba1: control = 0.34% (IQR: 0.08–0.63%) vs AD = 0.29%(IQR: 0.13–0.68%), P = 0.788; CFL1: control = 0.15% (IQR: 0.03–0.038%) vs AD = 0.08% (IQR: 0.04–0.022%), P = 0.314; P2RY12: control = 0.00% (IQR: 0.00–0.55%) vs AD = 0.09% (IQR: 0.00–1.09%), P = 0.290; CORO1A: control = 0.21% (IQR: 0.10–0.52%) vs AD = 0.17% (IQR: 0.03–0.33%), P = 0.406. However, there was a significant increase for Iba1 and P2RY12 in iAD compared to AD cases: Iba1: AD = 0.29% (IQR: 0.13–0.68%) vs iAD = 0.87% (IQR: 0.40–1.11%), P = 0.036; P2RY12: AD = 0.09% (IQR: 0.00–1.09%) vs iAD = 0.60% (IQR: 0.50–1.08%), P = 0.041 (Fig. 3). No significant difference was found in CFL1 and CORO1A loads between the AD and iAD groups: CFL1: AD = 0.08% (IQR: 0.04–0.022%) vs iAD 0.13% (IQR: 0.06–0.41%), P = 0.314; CORO1A: AD = 0.17% (IQR: 0.03–0.33%) vs iAD = 0.20% (IQR: 0.03–0.61%), P = 0.406).
We then explored whether the expression of the microglial motility-related proteins was correlated with the proteins that aggregate in AD. In the control group, correlations were detected for Iba1 and P2RY12 with (i) pan-Aβ (rs = 0.440, P = 0.046 and rs = 0.443, P = 0.042; respectively) and (ii) Aβ42 (rs = 0.691, P < 0.001 and rs = 0.629, P < 0.001; respectively) (Table 2). No correlations were observed with pan-Aβ and Aβ42 in the AD or iAD groups, and with ptau for the three groups.
We also assessed whether the different mechanisms involved in motility (actin-related or purinergic signalling) were related. In the control group, Iba1 was correlated with CFL1 (rs = 0.502, P = 0.025) and P2RY12 (rs = 0.572, P = 0.005). In the AD cases, Iba1 was related to CFL1 (rs = 0.559, P < 0.001), CORO1A (rs = 0.436, P = 0.014) and P2RY12 (rs = 0.558, P < 0.001). CFL1 was also correlated with CORO1A (rs = 0.530, P = 0.002) and P2RY12 (rs = 0.542, P = 0.001), and CORO1A with P2RY12 (rs = 0.357; P = 0.049). No correlation was detected in the iAD group (Table 3).
To explore the relationship between microglial motility and the neuroinflammatory environment, the MSD platform was used to measure the levels of IFNγ, IL1β, IL2, IL4, IL6, IL8, IL10, IL12p70, IL13, TNFα, IL1α, IL5, IL7, IL12/IL23p40, IL15, IL16, IL17A, GM-CSF, TNFβ and VEGF in the three groups. Significant increases in the AD group compared to the control group were observed for: IFNγ (P < 0.001), IL10 (P = 0.003), IL12p70 (P < 0.001), IL13 (P < 0.001), IL2 (P = 0.002), IL4 (P = 0.007), TNFα (P < 0.001), IL15 (P = 0.024), IL16 (P = 0.020) and TNFβ (P = 0.009). In the iAD group compared to AD, IL13 (P = 0.017), IL8 (P = 0.023) and VEGF (P < 0.001) were significantly increased; whereas IL7 was significantly decreased (P = 0.012). Of note, of the proteins that were significantly different between AD and iAD, IL13 and VEGF were also significantly increased with respect to the control levels (IL13: P < 0.001; VEGF: P = 0.012), while IL7 was significantly decreased (P = 0.003) (Table 4).
Evidence from animal studies shows that microglia are highly motile cells [14, 16, 34, 49], with motility impaired in AD models [6, 23]. This highlights motility as a functional aspect of microglia relevant to disease pathogenesis. Our aim was to characterise the expression of motility-related microglial proteins in the human brain and to evaluate its changes in AD and after Aβ immunotherapy. In the present study, we observed increased expression of the microglial homeostatic markers Iba1 and P2RY12 in the immunised group, which was associated with a less pro-inflammatory environment than observed in AD. We acknowledge that this approach has limitations, partly inherent to any post-mortem study, which reside in the absence of exploration of the temporal relationship between the different markers investigated, and with the analysis limited to the late-stage consequences of AD. Additionally, other proteins such as actin, the Arp2/3 complex, or the integrins also relevant for motility have not been evaluated here, as our investigation focuses on microglial-specific motility-related proteins. The strength of the study is in the examination of AD itself, rather than an experimental model of the disease which does not represent the whole extent of the complexity of the human disease. Specifically, most rodent models lack tau pathology, the generation and propagation of which microglia are proposed to have an important role. The novelty of our study resides in the combined quantitative assessment of several markers associated with microglial motility in relation to the neuroinflammatory environment, and the accumulation of Aβ and ptau. This study also takes advantage of the exceptional chance to explore the effect of Aβ42-immunotherapy which constitutes an important topic of study since there is currently substantial resource being invested in this therapeutic approach.
Aβ and tau pathology
As expected, the expression of the key features of AD neuropathology, Aβ and ptau, was increased in AD. The effect of the immunotherapy on AD has been previously reported by us [4, 32, 33, 42]. However in this study, we were able to compare the effects of the treatment with both controls and unimmunised AD. Interestingly, we observed that Aβ42 immunotherapy reduced the overall Aβ levels to those of controls, indicating an effect not restricted to the Aβ42 form. This is consistent with our studies on the same immunised cohort using antibodies specific for the Aβ40, Aβ42 and Aβ43 species and reporting significant decrease of the three species [21, 32].
Of note, its effect on the Aβ42 form seems to be more pronounced, with a reduction to significantly lower levels than those of controls. This highlights a possible higher affinity of the immunisation towards the Aβ42 species, perhaps due to the fact that full-length Aβ42 was the immunogen originally used in the AN1792 trial, even though earlier studies showed that the antibodies produced by the patients were primarily N-terminal specific . This implies that the antibodies are able to target all different species of Aβ. The ptau load was reduced to the level of the controls, confirming the effect of Aβ-immunotherapy on the tau pathology [4, 33]. Indeed, our previous studies on the immunised cohort showed that ptau load was reduced after Aβ immunisation, in association with a lower expression of the tau phosphorylating enzyme GSK-3b . Ptau load is mainly constituted of tau-containing dystrophic neurite clusters and ptau in neuronal processes , rather than tangles. Nevertheless, despite the decrease in ptau load as assessed by quantification in specific localised neuroanatomical regions of cortex, most of the iAD subjects still had progressed to Braak stages V or VI, as evaluated by the anatomical distribution of neurofibrillary tangles throughout the brain areas. We interpret these findings as indicating that localised removal of Aβ following Aβ immunotherapy results in a corresponding removal of ptau in that location, consistent with the amyloid cascade hypothesis. However, it does not stop the spread of ptau through the brain, consistent with the evidence for prion-like spread of ptau [4, 33].
Microglial motility-related proteins
In AD, expression of all four motility-related proteins examined was not modified compared with controls, consistent with another post-mortem AD study . In this paper, they evaluated expression of Iba1 and P2RY12 and also found no difference in the expression of both proteins between the control and AD groups. Interestingly, in that study, when the AD cases were split according to the Braak stage, a decrease for both Iba1 and P2RY12 proteins was observed in the late-stage of the disease (Braak stage V/VI) but only in the hippocampal area, a region heavily affected by neuronal loss and tau accumulation . In our study, even though most of the AD cases correspond to Braak stages V and VI, we did not observe decrease in these proteins, which could be indicative of regional differences between the hippocampus and the cerebral cortex due to the exacerbated degree of neuronal death and tau aggregation in the hippocampal region in late stages of the disease. With respect to CFL1, our observation confirms previous reports of no difference in CFL1 expression in AD . The presence of CORO1A has been observed in human microglia , but to our knowledge has never been assessed in AD.
After Aβ immunotherapy in AD, a 3-fold increase in Iba1 expression and a 6.6-fold increase in P2RY12 expression was observed. Data on anti-Aβ antibody titres measured in the blood during life some years earlier were available  and did not correlate with the microglial analyses. Iba1 is expressed by microglia in both physiological and disease conditions, suggesting that Iba1 participates in both baseline and directed motility. In vitro studies have characterised the role of Iba1 in relation to membrane ruffling, and the formation of phagocytic cups is Iba1-dependent . Therefore, the higher Iba1 expression observed is in accordance with the reported increased phagocytic activity associated with Aβ clearance  as the consequence of immunotherapy stimulating microglial phagocytosis towards Aβ. Thus, after immunotherapy, Iba1 expression is likely to reflect the motility changes associated with phagocytosis. Similarly to Iba1, P2RY12 is detected on all microglial cells and a role of P2RY12 in directed motility is also well-documented. The release of ATP to the extracellular space, caused by cell death, and its subsequent hydrolysis to ADP activates P2RY12 . Studies have shown that microglia from mice lacking P2RY12 expression show normal baseline motility but impaired directed motility [15, 26]. Moreover, it was observed that P2RY12 expression in ramified microglia was markedly decreased as cells turn to the reactive/amoeboid form [15, 38]. This suggests a role of P2RY12 in directed motility taking place at an early stage of the cell activation. Thus, P2RY12 expression after immunotherapy might suggest an increase both in baseline and directed motility. This is also consistent with the neuroinflammatory environment observed in the immunised group, which is slightly leaning towards a more anti-inflammatory profile, considering evidence that P2RY12 is downregulated under pro-inflammatory conditions and upregulated in an anti-inflammatory context [22, 30]. Of note, a previous study by our group in the immunised group showed that neuronal loss is correlated with Iba1-positive microglia and phagocytic CD68-positive microglia associated with pro-apoptotic neurons, but without accelerated cognitive decline compared to non-immunised AD patients . This suggests that immunotherapy may enhance microglial elimination of damaged and dysfunctional neurons. Considering the role of P2RY12 as a purinergic receptor upregulated with the release of ATP that occurs during cell damage, it could be hypothesized that the increased P2RY12 expression in the immunised cohort reflects microglial increased response, as a result of the treatment, towards damaged neurons.
Motility-related proteins and the pathology
Relationships between the microglial motility-related proteins and Aβ and ptau, key pathological features of AD, were explored. In controls, Iba1 and P2RY12 were associated with Aβ, with these associations lost in AD and not restored by the immunotherapy. It is recognised that with ageing, Aβ accumulates independently of the dementia status . Therefore, the relationship of Iba1 and P2RY12 with Aβ in the absence of dementia may reflect a healthy chemotactic microglial response towards Aβ, via the directed motility mechanism, as previously reported in the Cognitive Function and Ageing Studies group . In AD, these correlations were lost, possibly due to either (i) Aβ over-accumulation to a level at which microglia are overwhelmed and no longer capable of responding accordingly, (ii) microglia attracted to plaques and been immobilised there , or (iii) microglia being “distracted” by the tau pathology . However, the absence of correlation of the motility-related proteins with Aβ or ptau in the AD group is in favour of a microglial response towards other pathological changes, such as the synaptic/neuronal loss. Of note, the correlations were not restored after immunotherapy despite Aβ levels as lower as in the controls and this despite the increased Iba1 and P2RY12 expression. No relationship was observed with CFL1 or CORO1A.
Since no correlations were found between the microglial motility-related markers and ptau, this implies that the motility response driven by the proteins studied is independent of tau, or at least they do not directly interact with tau.
Interactions between motility-related proteins
Correlations between the microglial motility-related proteins were also investigated. In controls, Iba1 correlated with CFL1 and P2RY12, reflecting an interaction between the actin-related motility mechanisms and the purinergic signalling pathway in physiological conditions. Interestingly, in AD all markers were related to each other, implying that the pathogenesis elicited a coordinated response of the microglial motility proteins as part of the pathogenesis spreading. In the immunised group, no relation was observed despite the increased expression of the homeostatic markers Iba1 and P2RY12. This implies that immunotherapy did not restore the physiological profile of microglial protein interactions.
Inflammation-related proteins were evaluated to characterise the microenvironment. In AD, 10 were increased, including the pro-inflammatory proteins IFNγ, TNFα, TNFβ, IL12p70, IL2, IL15 and IL16, and the anti-inflammatory proteins IL4, IL10 and IL13. This reflects that both pathways of activation coexist in human AD as previously reported [9, 55].
After Aβ immunotherapy, only four markers were modified when compared with unimmunised AD, with increased expression of the pro-inflammatory chemokine IL8, the anti-inflammatory cytokine IL13, the vascular endothelial growth factor (VEGF), and a decrease in the pro-inflammatory cytokine IL7. IL13 and IL7 expression tends to reflect a slightly less pro--inflammatory environment after the treatment. VEGF expression was the most affected by immunotherapy with a 5-fold increase. This might be the result of (i) increased soluble Aβ species following plaque solubilisation  leading to neurotoxicity and angiogenesis via a VEGF-mediated mechanism , or (ii) reflected the vascular side effects reported after immunotherapy, known as amyloid-related imaging abnormalities (ARIA) including microhaemorrhages and oedema [5, 43]. Interestingly, both VEGF and IL8 are involved in angiogenesis . It is worth noting that, from the four markers that differed between AD and controls, IL13 and VEGF were also significantly increased with respect to controls, while IL7 was decreased with respect to control, emphasizing that Aβ-immunotherapy, even though inducing a slightly more anti-inflammatory environment, did not restore the normal state.
In conclusion, our study supports the experimental observation that microglial motility is impaired in AD, as evidenced by the changes observed in the interactions of the motility-related proteins with each other and with Aβ. As part of the AD pathogenic process, microglia move from baseline to directed-motility, involving all investigated proteins (Iba1, CFL1, CORO1A and P2RY12). Aβ immunotherapy seemed to result in increased microglial baseline motility (suggested by increased Iba1 and P2RY12 expression, which are considered “homeostatic” microglial markers), potentially due to decreased Aβ and/or tau, but without restoration of the physiological status of microglia.
Availability of data and materials
The data used and/or analysed during the current study are available from the corresponding author on reasonable request.
Amyloid-related imaging abnormalities
Bicinchoninic acid assay
Glycogen Synthase Kinase
Ionized calcium-binding adaptor molecule 1
Intercellular adhesion molecule
Purinergic receptor P2Y12
Region of interest
Transforming growth factor
Tumour necrosis factor
Vascular endothelial growth factor
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We would like to thank Dr. Laura Palmer at the South West Dementia Brain Bank (SWDBB), which is supported by BRACE (Bristol Research into Alzheimer’s and Care of the Elderly) and provided the control and AD cases. Multiple sclerosis samples were supplied by Dr. Djordje Gveric from the Parkinson’s UK Brain Bank funded by Parkinson’s UK, a charity registered in England and Wales (258197) and in Scotland (SC037554). We thank Elan Pharmaceuticals for the access to phase I AN1792 treatment study data and are grateful to all patients and carers who took part in the study. Our thanks also include the staff from Cardiff, Swindon, Oxford, Bath and Bristol Centres involved in the AN1792 study. We acknowledge the Histochemistry Research Unit and Biomedical Imaging Unit of the Faculty of Medicine, University of Southampton, that facilitated tissue processing, staining and analysis.
DKFB is funded by a CONACyT scholarship (270152 / 438262) and a University of Southampton scholarship (VCFoM/BocheD/TBC/170). Alzheimer’s Research UK (ART/PG2006/4, ART-EXT2010–1, ARUK-EG2015A-4) supported the collection of the immunised AD cases.
Ethics approval and consent to participate
The study was covered by the following ethical approvals: (i) the South West Dementia Brain bank (NRES Committee South West Central Bristol, REC reference: 08/H0106/28 + 5 for the controls and AD cases; (ii) the Southampton and South West Hampshire Local Research Ethics Committees (reference: LRC 075/03/w) for the iAD cases; (iii) The Multiple Sclerosis Society of Great Britain and Northern Ireland and Parkinson’s UK (reference 08/MRE09/31 + 5) for frozen tissue used as positive controls for the cytokine assay.
Consent for publication
The authors declare that they have no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Table S1. Demographic, clinical and post-mortem characteristics of the control group. Table S2. Demographic, clinical and post-mortem characteristics of the Alzheimer’s disease group. Table S3. Demographic, clinical and post-mortem characteristics of the immunised Alzheimer’s disease group.
About this article
Cite this article
Franco-Bocanegra, D.K., George, B., Lau, L.C. et al. Microglial motility in Alzheimer’s disease and after Aβ42 immunotherapy: a human post-mortem study. acta neuropathol commun 7, 174 (2019). https://doi.org/10.1186/s40478-019-0828-x
- Human microglia
- Cell motility
- Alzheimer’s disease

domain: biology


Women’s Self-Defense Class in Traverse City Teaches Personal Survival Skills
The American Taekwondo Association Martial Arts in Traverse City is hosting a class to make you feel safe in your surroundings.
A women’s self-defense course is taking place tonight at six!
Self-defense is an essential tool to have and ATA Martial Arts in Traverse City is getting you ready.
“Everybody has a possibility of being attacked,” Chris Pline, Owner of ATA Martial Arts in Traverse City.
Chris knows the importance of a women’s self-defense seminar and he welcomes anyone to the class tonight.
“It’s important the community knows how to defend themselves in case anything happens,” Pline says.
Amanda Corwin is a member of the gym and has done multiple self-defense courses here.
“It’s one of my favorites because some women feel intimidated to do a class with a man so with the women’s self-defense you get a chance to just go against people you might feel comfortable with first,” says Corwin.
They run through real life scenarios at this class with one of them being a hostage situation.
“I feel more confident walking to my car and I know the proper way to present myself in public as well,” Kelly Bator, gym member.
Learning the techniques and skills of personal survival to avoid being a victim.
“I never walked the streets feeling like something was going happen but now I definitely don’t I just walk around with self-awareness and a feeling like don’t mess with me,” Kelly Watrous, gym member.
“You never know when somethings going to happen. It’s better to have taken a course and have something in your back pocket that you can use that hopefully you’ll never have to use versus getting into a situation and freeze and not know what to do,” says Watrous.
For more information on this course head to the ATA Martial Arts website for the registration link!


text:
She scribbled violently over the piece of paper, frustrated and annoyed by her own frustration. She started shaking and her fingers were cold, the rest of her body aching and sweaty. She bit her lip so hard that she nearly drew blood, but stopped herself and tried to calm her quickening breath. There was absolutely no need for this reaction, and she knew it. It was just one of those days that she hated; where everything started bubbling up inside of her, resisting her own resistance.
"Done Rhyli?" A passing teacher asked her, picking up the piece of paper that lay face down on her desk.
"Mmhmm." She murmured idly. To everyone around she looked perfectly calm, normal and controlled.
That was the truth, as anyone else was concerned. Even she herself believed it at times, but now she threw all of that away, except the mask that hid the anxiety building up inside her.
She ran a hand through her blonde hair, silently savoring the pain that bit at her scalp as her fingers caught a few knots. She needed to calm down, and she knew that, but knowing it and accomplishing it were two completely different things. She closed her eyes and began to doodle in her notebook. She took a shaky breath and began to calm herself, for now.
She wasn't some psychotic depressed 15-year-old. She didn't cut, or mutilate herself in any way. She didn't drink alcohol, she didn't do drugs, and she didn't even smoke. To the outside world she was the nearly perfect teenage girl, or at least at first. She was mouthy, and a smart ass, and took great pride in that, though she knew she shouldn't. Her motto was, "If you want my respect you need to be the first to give it." Certain people, her father for example, did not agree with this. She mouthed off to teachers at school, and amused herself by deliberately doodling over her notes when she knew the teachers hated it. And, the best part about it, she didn't care.
Her outfit matched her attitude. She dressed punk, with the bracelets and chains on her baggy jeans. An armband adorned her right arm, with little designs across it. A thick black and silver belt barely held her pants up and she wore a simple black tank top and jacket overtop. It wasn't outrageous, but it was enough to make the statement she wanted.
She was just a typical, 15-year-old girl in high school, dealing with typical high school drama with her typical high school friends. Only, this wasn't entirely true. Her 'typical' friends, each had issues of their own, and each and every one of these friends had her phone number, which they each used frequently, one in particular more than most.
Rhyli dealt with most of these problems easily. Normally her friends were just too worked up to think clearly. Other times though, her friends' problems built up over her own. She didn't really have too much trouble with her own problems, or at least she didn't think she did. Her father and she fought frequently, and often times it got to the point where she could only leave. She'd pick up her bag, always holding her CD player and her wallet in case she needed it, and then she'd grab her dog's leash, using the always-dependable excuse "I'm taking the dog for a walk." And so she'd walk; Sometimes for a few minutes, sometimes for hours, more frequently the latter of the two options.
Today was just one of the days where her thoughts built up and the stress was tugging at her, causing her to lose her concentration and thus getting frustrated, wanting only to throw something across the room to express her frustration, but that clearly wasn't an option.
She walked the halls now, a bag slung over her shoulder and a few things gathered in her arms. She looked down, struggling with the clasp of her bag as she attempted to put her things into it. Suddenly then, a pair of hands grasped her shoulders and an unfamiliar face bent to meet hers, startling her.
"Who ARE you?" He asked as though this very thing had been bothering him for weeks.
"I'm...uh..." She stammered, trying to think. "A little freaked out..." She managed to say, looking at him like he was insane.
"Right, well then, you're sitting with me at lunch today. Third table from the back, center. Ok?" He said quickly, straightening up.
"Uh." She blinked rapidly.
"Ok, good!" He said, suddenly acting as though he was in a very chipper mood. "See you then." He called over his shoulder as he walked away.
"What?" She said to no one, extremely confused by this point.
The only thing she got out of that encounter was that a punk kid, who just happened to be rather attractive, had just declared that she was eating lunch from him. "Third table from the back, center." She repeated to herself, on autopilot to her next class.
During the next two classes she had to endure, she kept running over that scene in her head. From what she remembered, the boy was older than she was, probably a grade or two higher, he was punk, with an amazing necklace with two nails crossed and worn as a choker, and he was pretty damn hot, but so were a lot of guy she knew.
As she stood in line for her lunch, she glanced over toward the table the boy had indicated, and saw him gazing up at her, smiling. She nearly choked as she turned back towards her friends, debating whether or not to sit with him.
"Um, Chris…" She said to her friend, still thinking. "I uh…"
"She's sitting with us today." She heard him behind her now, grasping her shoulders. She looked up to see him gesturing towards the table.
He steered her toward his table, while she walked somewhat nervously with him. He pushed her toward the bench. "Sit" he commanded.
She gave him an annoyed look, but sat.
"So, this is lil' miss freaked out huh, Kai?" the boy beside her asked. He was wearing more color than she knew could be combined in a single outfit. Tie-dye tee-shirt, green pants, and his hair was dyed a bright purple. He even had a dozen or so earrings, all different colors.
"Yeah, and you're the king of color." She snapped, and then turned back to the first boy she'd met. "Rhyli."
"Malakai." he replied, nodding and smirking at the way she'd spoken toward his friend.
"Kaiiiii!" The other boy whined beside her. "She made fun of my pretty colors!"
Rhyli laughed along with the rest of the guys. There were four other guys, besides Malakai.
The first was Liam. He was about 17, had long black hair, and brown eyes, pierced ears and looked almost gothic. He was pretty quiet and seemed almost depressing, but after a while she realized he really was a happy person.
Next was Cole. He was 16, had longish brown hair and brown eyes, and looked like the most normal out of everyone, except that he had so many piercing it was hard to talk without staring at them.
Then there was Brody, the oldest at 18 with long curly brown hair and eyes. He dressed gothic, wore spiked bracelets and a chain necklace, and had his ears and lip pierced. He really was the moody one of the bunch. He automatically seemed not to like Rhyli, but soon he stopped his glaring and kept talking like the rest of them.
And next was 'The king of color' publicly known as R.B., which, coincidentally stood for Rainbow boy. Malakai stated that no one really knew his real hair color and R.B. refused to tell, saying it was purple, and clashed with his bright green eyes. He was the youngest and most immature and obnoxious of them.
They all sat and chatted for a while. She was comfortable as she ever could be with people she'd just met.
"So, you've told me about everyone else Kai." She said, smirking. "But all I know about you is that you have a really odd way of making a good first impression."
They laughed at this together, and Brody spoke first. "All you need to know about Kai is that he's pathetic and hasn't had a good girlfriend in years, and thus has gone way too long without getting laid."
They all laughed, but Rhyli only smirked. "Is that what I'm here for then?" She asked coolly.
"No way." Kai was the first to answer this one. "You were just hangin' with the wrong people, that's all. I thought you needed some relief from your girly psycho crowd over there."
She didn't deny this, but she secretly agreed with him. She just shrugged. "Okay then."
"What about you Rhyli?" Brody spoke again, looking suddenly interested.
"What about me?" She countered.
"What's with the psycho crowd you were hangin with?" He continued.
"Don't call them that." She said seriously. "And there's nothing 'with' them. They're just the people I ended up with after I got up here. Never really made an effort to find anyone else to hang with."
"And what's with this 'I'll kick your ass' reputation you got goin' round?" Cole asked, raising a pierced eyebrow.
"'Scuse me?" She asked.
"What're you a feisty one or what? I hear you'll kick anyone's ass if they piss ya off enough." He continued.
"Yeah?" She asked, shrugging. "So what?"
"It's true then?" Brody asked. "Lil' girly'll kick our asses."
"Call me 'lil' girly' again and I'll let you know." She said, staring him down. She had a way of doing that. Something in her eyes told everyone that she wasn't joking. It confused teachers, but it let anyone else know she wasn't kidding.
Malakai shook his head, smirking. "Blonde hair, blue eyes, gone punk instead of preppy." He laughed.
"Problem?" She asked, giving him the same look.
"Never." He said, smirking at her.
She let down her mask then for a moment. She noticed then that he had bright blue eyes that stuck out from behind his long black straggly hair.
"I take that back." He said suddenly. She raised an eyebrow. "Blue-green eyes."
She blinked and then smiled a little. "Okay then."
The next few days carried on about the same, for a while Malakai still had to steer her to the table, but she eventually started going on her own. She'd laughed when Kai stopped, in mid-rising to go get her, when she started walking towards him already.
The days were going well, despite the fact that her friends were starting to complain that she'd deserted them. She was calmer, and happier. She didn't have anything to hide from them like she did from her other friends. Or, that's what she was thinking until the night that her father came home, annoyed for some reason that she didn't bother finding out.
"Why the hell aren't you in the kitchen helping your mother clean?" He'd snapped when he'd seen her online, chatting with Malakai. "gtg." She typed quickly, and then signed off.
"I cleaned it this morning—"She began to explain.
"It doesn't matter if you cleaned it, it's filthy! Get your ass in there and help your mother!"
"Dad, mom said that someone else will do it—"
"Stop your backtalk and get it there!"
"No! Dad would you listen already?" they'd walked into the kitchen now, screaming at each other.
And then, Slap. Rhyli staggered and lost her balance, falling hard into the pantry door.
"Don't touch me!" She screamed without thinking, reaching a hand up to her cheek.
He hit her again. "I said don't you backtalk me!" and again. "Now clean the fucking kitchen!"
She regained her balance then and walked out the door, grabbing her bag and the leash. Her dog followed her quickly, seeing the leash and making it out the door before Rhyli left without a word, only the click of the door shutting behind her father's back. She never cried, never shed a single tear, and she never let herself get out of control.

domain: literature


Natasha Chansa Repudiate Joining XYZ Entertainments
Having started music at a young age, Natasha Chansa has been signed with Zed Arts and following up all her releases she always tags along her label, well things went sore after the act posted a picture posing with the XYZ Heavyweights inline with a caption “Zambian music at its finest , XYZ Entertainment Featuring Zed Arts Records , Tag some of your favorites from this picture !“.
Everything was lit, until a music fan gave out a conclusion stating, Princess Natasha Chansa should join the XYZ Family.
Check out the thread below.
|Rich Bizzy - Enjoyment
|Yo Maps – On The Low
|Macky 2 ft. Dimple Williams – System
|Kay Figo ft. Macky 2 – Taima Taiwa
|Yo Maps – Kamusango
|Y-Celeb ft. Chile Breezy - Alimpala
|Jemax ft. Y Celeb - Bwaka Musango
|Drifta Trek ft. Neo - Before
|Rich Bizzy ft. Nez long – One Life
|Namadingo - Sakaka
|Yo Maps ft. Slap Dee - Komando
|Bobby East ft. Kantu – Toxic