title: Nikita khnykin

text:
Nikita Khnykin was born in Norway in 2003 and started playing the piano at the age of nine. He is currently studying with Prof. Julia Mustonen-Dahlkvist at the Musikhögskolan Ingesund and he’s artist-in-residence at the Ingesund Piano Center in Sweden.
Since 2016, Nikita has also been a member of the talent program at Prof. Jiri Hlinkas Piano Academy in Bergen, a mentoring program where he regularly receives lessons from leading Norwegian musicians such as Leif Ove Andsnes and Håvard Gimse and attends master classes with world-class pianists such as Bertrand Chamayou, Richard Goode, Jan Jiracek von Arnim, Marianna Shirinyan and Polina Leschenko. He is also a scholarship holder of the International Music Academy in Liechtenstein and takes part in the intensive weeks and activities that take place there.
Nikita has won several important national and international competitions including several top prizes at the “Norwegian National Music Competition” where he won all sorts of prizes in 2020 such as the “Talent of the Year”, the “EMCY Prize” and the audience award. He also won the 2022 “Swedish National Music Competition Unga Solister”, the 2016 “Steinway Piano Competition” in Norway, the 2015 “Young Musician International Competition” in Estonia and the 2016 “Young Piano Competition” at the Prague Conservatory.
He has appeared as a soloist at concerts and festivals in Norway and abroad, such as the 2019 De Unges Konsert with the Bergen Philharmonic Orchestra conducted by Cathrine Winnes, where he received the Audience Award and a scholarship from Kavlifondet. Other highlights include several performances with the Norwegian Radio Orchestra (KORK) of Tchaikovsky's Piano Concerto No. 1 with Ari Rasilainen in 2018 and Prokofiev's Piano Concerto No. 3 in 2020, broadcast live on TV and radio, as well as solo performances with the Kaunas City Symphony Orchestra. Recent achievements include a tour with the Ingesund Piano Center in late March 2022 for Dubai Expo 2020, presenting works by Nordic composers to an international audience.
In fact, Nikita has also worked with the Oslo Philharmonic Orchestra, albeit as a composer. His work Poeme for orchestra was performed and recorded by the Oslo Philharmonic Orchestra in 2021, conducted by Christian Eggen.
A passionate chamber musician, Nikita has played in a variety of ensembles, from string duets to wind sextets. Together with the clarinet trio DaNiBi he was part of the Norwegian mentoring program Crescendo in 2016, in which he worked together with Konstantin Heidrich. Other important teachers are Lars Anders Tomter and Henning Kraggerud.
As a chamber musician, he has performed at the Kavli Awards, Bergen International Festival, Oslo Chamber Music Festival and numerous other high-profile occasions, such as for Queen Sonja of Norway. In 2022, he played Rachmaninoff's 2nd Piano Concerto with the Oslo Symphony Orchestra and was subsequently awarded the prestigious Levin Prize. In 2023, Nikita performed at the Bergen International Festival, the Trelleborg Piano Festival, the Lofoten International Chamber Music Festival and the Helsingborg Piano Festival, where he made his debut with the Helsingborg Symphony Orchestra.
Nikita received a handful of grants from The Kavli Trust (Kavlifondet), Ånun Lund Rejs Minnefond and Drømmestipendet.


tier: cub

shelf: civic-law

title: By Ayinke Adebayo

text:
By Ayinke Adebayo
Embattled Police Chief, Abba Kyari has claimed that the National Drug Law Enforcement Agency, NDLEA has been assisting drug dealers to bring in Cocaine into Nigeria.
Kyari made the claim in a N500 million fundamental right enforcement suit marked FHC/ABJ/CS/182/22.
The disgraced police officer accused the anti-drug agency of assisting drug peddlers to ship in Cocaine into the country. He also stated that he was ‘framed’ by a disgruntled officer of the agency after he went to demand compensation for the whistleblower that gave information that led to the arrest of a suspect.
An affidavit filed by Kyari’s brother, One Mamman Nur, rehashed the claim stating that drugs were being shipped in from Ethiopia with the ‘knowledge of NDLEA officers’.
Narrating the entire situation in the suit, Kyari stated; “That on that fateful day, the suspect as usual shipped drugs (cocaine) from Ethiopia to Enugu Airport. The officer of NDLEA cleared him within the airport, then officer of FIB-IRT,on intelligent information arrested the suspect.
“That before the informant accepted to work with the officers of FIB-IRT, they both had an agreement to compensate the informant.”
“That officers of FIB-IRT, having noticed that officers of NDLEA were the ones clearing the said suspect with his drugs (cocaine), they immediately took the suspect to NDLEA and also reported those officers involved in the act.”
“That instead of the NDLEA to handle the matter accordingly and bring their officers involved to book, they decided to frustrate the informant, thereby refusing to compensate the informant.”
“That it was at this juncture that the attention of the DCP Abba Kyari was called, who because the NDLEA officer involved is his friend, decided to intervene for the purpose of compensating the Informant.”
“That the NDLEA officer who refused to compensate the informant was the one who framed DCP Abba Kyari up on a trump-up allegation that he was trying to bribe him.”
“That the agent of the Respondent declared the Applicant wanted because of a mere allegation that he was trying to bribe an NDLEA officer without sufficient proof.”
“That the Nigerian Police Force, based on the allegation of the Respondent arrested the Applicant on February 12, 2022, and handed over to the NDLEA.”
Kyari’s bail hearing comes up on March 28th at a federal high court in Abuja.


shelf: science-engineering

title: Simplex modular cleanrooms relocates in the us

text:
Subzero Engineering, the global leader in turnkey engineering solutions for data centers, industrial cleanrooms and mission-critical environments, has announced a new 155,000 sqft facility in Salt Lake City, Utah.
The extensive premises enables the company to expand its capabilities while enabling the company to relocate its Simplex manufacturing facility into the same premises.
The new headquarters is set to transform the industry with its cutting-edge amenities. The new facility comprises 95,000 sqft of dedicated manufacturing space, 25,000 sqft for shipping, receiving and storage, and a further 35,000 sqft of office space.
This strategic move, coupled with recent expansions in the sales, engineering, manufacturing and installation teams, positions Subzero Engineering for remarkable growth in the coming years.
The new facility, located at 805 South 3600 West, Salt Lake City, Utah, represents a significant milestone for the company and the industry as a whole. This new facility will provide the space needed to support the combined teams of Subzero Engineering and Simplex Modular Cleanrooms, one of the most prominent names in modular cleanrooms, softwall curtains, strip doors, separation and process isolation, and their growing customer base. It will also encourage and develop the collaboration and teamwork of both teams to assist customers with custom projects, product enhancements, and new products while delivering support for all cleanroom and separation needs.
This new facility will provide the space needed to support the combined teams of Subzero Engineering and Simplex Modular Cleanrooms
With an entire area dedicated to research and development, the company will be in a pivotal position to progress innovation, design and productivity. This is expected to become a prominent resource, establishing the company as a key partner in the evolution of the industry.
Expansive Product Demo Room: The product demo room showcases the full range of the organisation’s data center and cleanroom products, allowing customers to comprehensively explore their options. The new product demonstration room is testament to the company's commitment to showcasing its cutting-edge offerings.
Cross-Product Knowledge Sharing: The new facility fosters collaboration and knowledge sharing between the data center and cleanroom product lines.
Research and Development: The dedicated Research and Development area will be the company’s epicenter of innovation, driving the development of cutting-edge solutions.
Enhanced Inventory and Materials Storage: The larger space allows for expanded inventory stock and improved materials storage capabilities, ensuring timely deliveries and better service.
Improved Customer Support: With the additional personnel and facilities, Subzero Engineering is committed to providing even better customer support and assistance.
Central Shipping Hub: Salt Lake City's strategic location as a central shipping hub will expedite shipping and receiving processes, benefiting customers nationwide.
Shane Kilfoil, President of Mission Critical Environments, covering both Subzero Engineering and Simplex product lines said: “We've gone from starting in a garage, to now having a 155,000 sqft facility and seven different production lines. We've also tripled our workforce to help keep up with demand over the past 18 months. We foresee that continuing — but we also recognise that not everything can be done just with people. So, we’re making active investments in machinery and automation that will help us to further keep up with demand and take the strain off our employees to allow them to focus on other more value-added offerings and solutions that we can give to the customers and end users.”


shelf: software-docs

title: Spot the ant stop the ant month 2017

text:
HAVE YOU TESTED YOUR YARD FOR LITTLE FIRE ANT? It’s soooo easy and even more important. Find them…before they find you!!
We need your help! ...you can pick up free test kits at these Oʻahu locations to collect and send in your ant samples:
Mānoa Public Library: 2716 Woodlawn Dr. Manoa, HI 96822 HOURS
Waikiki-Kapahulu Library: 400 Kapahulu Ave. Honolulu, HI 96815 HOURS
Waimānalo Library: 41-1320 Kalanianaeole Hwy. Waimanalo, HI 96795 HOURS
Pearl City Home Depot: 1021 Kamehameha Hwy, Pearl City, HI 96782 HOURS
Do you need a free test kit? NO…all you need are chopsticks and peanut butter: How To Test for LFA
Q: We’ve already got fire ants are everywhere…I always get stung at the beach and the park. What’s the big deal?
A: Most people on Oʻahu have experienced “Tropical Fire Ant”…but “Little Fire Ant” is a different beast. Tropical Fire Ants live in sunny, dry areas….while Little Fire Ants like it shady and damp. Also, Little Fire Ants will live in the trees and on the ground. People who have experienced areas heavily infested with little fire ant talk of “raining ants” and getting repeated stings on their head, face, arms and back. This is the situation we want to avoid on Oʻahu.
Q: I’ve never seen little fire ant at my place, why should I test?
A: Just because you don’t see them, doesn’t mean they’re not there. Little fire ant only spread about 60 feet per year (as long as we don’t move them). So, they could be hanging out in a shrub, going unnoticed for a long time. We want to find them BEFORE they spread all over your yard. If we can find them while their colonies are still small…it’s so much easier to eradicate them.
Q: What happens if I do have little fire ants?
A: Because they are still not widespread on Oʻahu, Hawaiʻi Dept. of Agriculture will help you get rid of them!
Q: Do I have to have go pick up one of these fancy test kits?
A: Nope!! All you need are some chopsticks, popsicle sticks, or coffee stirrers, a ziplock bag and some peanut butter (use lunch meat or mayo if you’re allergic to peanut butter). Testing is quick and easy in just 5 steps! For testing details, CLICK HERE
For more information on little fire ants in Hawaiʻi…visit: www.stoptheant.org


shelf: health

title: New 988 hotline is the 911 for mental health emergencies

text:
A new three-digit number that connects callers directly to the National Suicide Prevention Lifeline went live on Saturday nationwide.
The line will connect callers to trained counselors who will listen, provide support, and work to understand the mental health problems these callers are experiencing, and connect them to the right resources.
Today, an estimated 8% of all calls to 9-1-1 are related to a mental health crisis, according to data from Vibrant Emotional Health. The new 9-8-8 number will provide an easy-to-remember three-digit number people can call to receive the right mental health crisis support.
It’s no secret that people with mental illness don’t get the help they need when they experience a crisis. The National Suicide Prevention Lifeline has proven effective since its establishment in 2005. The three-digit number 9-8-8, similar to 9-1-1, will make it even easier for folks across the country to connect with help near them.
The Lifeline has a network of counselors at local crisis centers who answer the contacts the Lifeline receives every day. Numerous studies have shown that callers feel less suicidal, less depressed, less overwhelmed, and more hopeful after speaking with a Lifeline counselor.
In Alabama, the 9-8-8 number has provided an opportunity to create a commission whose sole purpose is to ensure crisis services are available and adequately funded to serve people in this state.
The new system is expected to reduce health-care spending with more cost-effective early intervention, reduce the use of law enforcement and other safety resources by diverting calls from people in mental-health crisis, meet a growing need for crisis intervention and help end the stigma people feel when seeking mental-health services.


shelf: finance

title: Read the article of July 18, 2023

text:
Read the article of July 18, 2023
There has been a sharp slowdown in recent months for the real estate market of second homes in Umbria. Paola Berlenghini, real estate agent member of the supervisory committee of the Umbria real estate exchange, describes the situation: "In the first 9 months of 2022 it confirmed a good performance. The brick symbolically represents stability precisely in periods of uncertainty and precariousness. The house claims its role as a safe haven par excellence. In recent months there has been a slowdown which has lasted until the second half of 2023 due to inflation which contributes to cooling the positive momentum that has affected the real estate sector in recent years". The forecasts for the prices of properties to be renovated in the immediate future: "On the one hand they are undergoing declines, on the other, thanks also to the redevelopment policies of the real estate assets and a limited offer of renovated houses, for the second half A revaluation of prices is expected in 2023, estimated by some to be a few percentage points for each leap in energy class achieved thanks to real estate regeneration". The market is now much more targeted: "We are looking for quality, efficient and renovated houses. The offer has dropped numerically and has improved qualitatively. Rents have seen a slight increase: for furnished two-bedroom apartments in the historic center , the average values fluctuate between 500 and 800 euros per month, while for houses in the countryside, with up to 3 bedrooms, furnished, between 800 and 1400 euros per month, in addition to the payment of all ancillary charges". Umbria always remains a destination of great interest for those who have to invest in a second home: "Man remains fascinated by nature and simple things, by houses not altered in their typicality and with the possibility of a view overlooking natural elements ". On the Umbria Chamber of Commerce website it is possible to monitor the quarterly price lists divided by Municipality.


shelf: education

title: Upgrade your dreams with gradding

text:
New Delhi (India), June 5: With 15k+ enrollments, 98% visa acceptance rate and tie-ups with over 700 institutes across the world, Gradding is among India’s top ed-tech company. It was founded in year 2012 and elevating itself with time and technology. That is how it is also among the India’s first AI-powered study abroad and test preparation platform where the aspirants get A-Z services, right from university selection to finding an ideal accommodation overseas.
Gradding is a one-stop solution for all the needs of aspiring students. It’s been a decade of Founder, Mamta Shekhawat along with her team helping the aspirants to mould their future. Let us know the founder’s views who has dedicated herself in making this dream come true.
Q1) First, Would You Tell What Services Gradding Provide?
Gradding stands for growing, responsive, accountable, dynamic, distinguished, illuminating, nurturing, genuine. Hence explained! It is expanding every day and is accountable for the services. The experts are experienced and certified. Moreover, their guidance helps the aspirant for their dream journey.
Coming towards services, we cater to all the needs right from institute and country selection to settling down in the host country. We also help the students to prepare for the English language tests like IELTS, PTE, TOEFL, and DET along with French and German languages with the certified trainers and mentors. Also, the accommodation assistance is given for a comfortable stay in the host country.
Q2) From Where Did You Get Inspiration to Start Such a Platform?
I started my career with prestigious MNCs like Hitachi & ITC in different roles across business sectors like employee engagement, business development, and talent management. Later, I joined Cognus Technology as the vice- president. Then I realized the need of a unique platform that caters to the aspirant’s problem. A place where students can invest trust over time and navigate through all the hurdles.
Q3) There Might be Instances When the Demands and Expectations of Aspirants are Much Higher Than What They Can Achieve. How Do You and Your Team Manage These Situations?
That one is a difficult thing to answer. However, aspirants indeed come to us with lots of expectations and we were established with the motive to cater all their needs under one roof. The major concern is the unwillingness of youth to indulge in research. They just hear something and run to us. For instance, Germany is popular for free education which is true but they have specific eligibility criteria like excellent academic records. But where the problem occurs, they just know the “FREE EDUCATION” but not the challenges behind it. Since Germany is a nation where English is applicable only in the academic world. I have seen many students returning to India from Germany just because they lack language skills.
That’s why we analyze the academic and financial profiles. Gather everything from their record to the country’s requirements and then we tell them the areas where they lack. In this way, their high demands are justified.
Q4) What if They Don’t Agree to the Solution Provided?
The various institutes frequently visit us. Recently, we had representatives from renowned universities like the University of Wales Trinity Saint David, Cardiff Metropolitan University, University of South Wales, and the University of Hampton. They analyzed the academic profile and gave personalized attention. These kinds of hybrid events help them to know better options and the future scope. More than 55 spot assessments were done. Also, 100+ study abroad aspirants attended this event.
Q5) What Message You Would like to Give to the Study Abroad Aspirants?
“Never think you’re late!” This is the message I would like to give to all the abroad aspirants along with all those who crushed their dreams due to various reasons. Your time starts as soon as you take a step forward in your career. Seek expert help and align your goal to mould the future in the desired direction.
If you have any objection to this press release content, kindly contact email@example.com to notify us. We will respond and rectify the situation in the next 24 hours.


shelf: home-craft

text:
A 10-Point Plan for (Without Being Overwhelmed)
Essential Insights right into Furnace Repair Work Services
When wintertime cools embeded in, having a dependable furnace makes all the difference in preserving heat and convenience in your house. Nonetheless, like any kind of other home appliance, furnaces can experience concerns gradually. Understanding the significance of furnace repair services can assist homeowners make more educated choices when confronted with heating troubles. In this post, we will delve into the vital elements of heating system repair work solutions, from indicators that your heater requires interest to picking the right service provider.
One crucial indication that your furnace calls for repair work is uncommon sounds. If your normally silent heating system begins to rattle, moan, or whistle, it’s a signal that something is amiss. These audios can point to issues such as loose parts, worn bearings, or other mechanical troubles that need professional focus. Additionally, if you notice inconsistent home heating or chilly places in your house, this variance often suggests that your heating system isn’t functioning efficiently and requires repair work.
One more element to take into consideration is climbing energy expenses. If your home heating expenses have actually suddenly surged without a corresponding boost in use, your furnace might be functioning harder than necessary. This inadequacy could stem from a range of concerns, including stopped up filters or ductwork issues. In such cases, looking for professional fixing services can not just restore your heating system’s effectiveness however likewise conserve you cash in the future.
Selecting the ideal heating system repair solution is important for guaranteeing your furnace runs smoothly. Try to find technicians that have a strong credibility, proper qualifications, and experience with your certain heating system design. Reading consumer reviews and asking for referrals can lead you to reliable companies. Furthermore, guarantee they supply a guarantee on parts and services, as this can provide satisfaction and assistance protect your investment.
In conclusion, being positive regarding heater maintenance and repairs can save you time, money, and discomfort during the colder months. By recognizing the indications of a struggling heating system and seeking expert assistance, you can ensure your home remains warm and inviting. Do not delay in attending to heating system issues; rather, rely on reliable repair service services to keep your heater in leading problem for several years to come.


shelf: history-culture

title: Story

text:
Synopsis/Plot : J Baby, the upcoming Tamil film starring actor Urvashi and Attakathi Dinesh in the lead roles.The film is written and directed by Suresh Mari in his directorial debut. The film is produced by filmmaker Pa Ranjith under his banner Neelam Productions.A story filled with emotions true to our home. J Baby will be a family drama with Urvashi and Dinesh expected to play mother and son duo. Apart from them, the film also stars Lollu Sabha Maran who was recently seen in Vadakkupatti Ramasamy alongside Santhanam. The film is expected to have comic, emotional and drama portions.
Has Good Moments, But Overly Stretched!
A relatable true story that is made as a watchable drama with a mix of family emotions and comedy. Long-drawn-out narration and old-fashioned presentation are the major issues, stronger technical aspects and a much crisper duration would have definitely made it a worthy film.(more)
Source: Ashwin Ram, MovieCrow
Urvashi anchors this inconsistent film with momentarily engaging moments
J Baby is proof that good acting performances as well as inconsistent and momentarily engaging scenes aren't enough to save a film.(more)
Source: Roopa, Times Of India
Urvashi is the mothership of this superficial film
the film had its tear-jerking moments with some audiences sniffling, but that momentary relatability cannot make a brilliant movie.(more)
Source: Priyanka sundar, Firstpost.com
Urvashi saves an otherwise overly melodramatic film
J Baby is more than a mother, more than a woman who showers love, more than an elderly person dependent on her kids during the last leg of her life, and more than a woman who keeps on giving. And we see that side of her only for sometime.(more)
Source: TNE, The New Indian Express


text:
Advanced Enterprise Content Management Powered by M-Files
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Looplex Content utilizes AI to automate the organization, classification, and management of your documents and emails. This smart system enhances knowledge and risk management, ensuring compliance, accuracy, and efficiency in handling vital content.
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Store frequently used clauses and claims in playbooks for easy insertion into documents. This feature streamlines the drafting process, ensures document consistency, and significantly reduces the time required to prepare legal documents.
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Looplex Content seamlessly integrates with popular tools like Microsoft Outlook, Office, and Google Docs. Whether created through Looplex Builder or manually within your productivity suite, all content is captured, centralized, and easily accessible.
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Version Control and Risk Mitigation
Ensure your team works from the most up-to-date documents, reducing the risk of miscommunication and errors. Looplex Content’s robust version control and tracking features keep teams aligned and minimize costly mistakes.
Customizable Workflows and Automation
Create tailored workflows to suit your organization’s specific needs. Automated workflows streamline approvals, collaboration, and document routing, improving efficiency and reducing manual errors.
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Secure sensitive information with granular access controls. Administrators can assign permissions at the document, folder, or metadata level, ensuring that only authorized personnel can view, edit, or share specific content. This ensures confidentiality and compliance with industry regulations.
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For more information or to request a demo of Looplex Content, contact our support team.


tier: polar

text:
Venus Express' Infrared Camera Goes Filming
- May 11, 2007
- European Space Agency
- An exciting new series of videos from ESA's Venus Express has been capturing atmospheric details of day and night areas simultaneously, at different altitudes.
An exciting new series of videos from ESA’s Venus Express has been capturing atmospheric details of day and night areas simultaneously, at different altitudes.
The south pole of Venus and the double-eyed storm permanently rule atmospheric phenomena in that area of the planet. They are key to understanding the global atmospheric dynamics on Venus and will contribute to a better comprehension of the global meteorology of the planet.
The south pole of the planet and its gigantic double vortex has been pictured as never before.
In the search for all possible clues on how to solve the global atmospheric ‘puzzle’, the team of scientists behind the Ultraviolet, Visible and Near-Infrared Mapping Spectrometer (VIRTIS) on board Venus Express, have tried something new – starting from the south pole.
They started by focusing on this target from the advantageous position of the orbit apocentre (the furthest distance of the spacecraft from the planet). This allows the instrument to keep the target in the field of view for longer than in other portions of the orbit, where the spacecraft travels faster. In this favourable position, scientists made efficient use of the multi-wavelength capability of VIRTIS.
By using wavelengths longer than 3 microns in the thermal infrared range, VIRTIS can obtain a combined view of the day and night sides simultaneously. This is more convenient since at shorter wavelengths, the difference between the thermal radiation emitted on the day and night sides is too high to observe both regions simultaneously without ‘blinding’ some channels of the camera.
“It is comparable to looking at bright, sun-illuminated snow and at a dark sky without having to change your glasses,” said Giuseppe Piccioni, VIRTIS co-Principal Investigator. “In addition, within this observation process, not only can we look at the dark and lit sides of the south pole at the same time, but we can also look into the atmosphere at different depths. What we are building is the most complete 3D data set of the Venusian atmosphere to date.”
The VIRTIS videos of the south polar vortex presented here are the result of combined observations at two different wavelengths (3.8 and 1.7 microns, respectively) used at the same time. The various images were taken over five orbits, during a time-span of about 8 hours per orbit.
The 3.8-micron channel was chosen because of its compatibility (in exposure time) with the 1.7-micron observations, as well as for its capability to provide information about the cloud deck at about 65 kilometres altitude over the planet. The 1.7-micron wavelength was chosen to probe the atmosphere below the clouds when looking at the night side.
It is clearly possible to see that the morphology of the vortex changes a lot during the 8-hour observation session and from one orbit to the next (one Venus Express orbit is 24 hours long).
It is interesting to note that due to ‘bad weather conditions’, by the time of the observations, the videos do not show the maximum achievable image contrast. In fact, the visibility of the polar structure was somewhat reduced by the local increase of the upper atmospheric haze.
“If the weather permits, by extending the time span of our future observations, we may have the chance to obtain even clearer and more detailed views of the polar vortex,” added Piccioni.
“With video sequences of this kind, combining all the pieces of information together, we can study the dynamics and the evolution of the vortex both in the short and the long term,” said Pierre Drossart, the other co-Principal Investigator on VIRTIS. “What we want to understand is the overall 3D thermal structure of the vortex, especially the vertical variation of the horizontal winds.”
The next step will be the correlation of this data and data collected in the next sessions, with fluid dynamics computer models. This will eventually help the scientists create the best possible atmospheric model of Venus to date.
Cite This Page:


title: Kubernetes an agile and secure option for smes to deploy the cloud 568264

text:
In just over a year, the COVID-19 pandemic brought about years of change in the way all companies in do business.
McKinsey says companies have accelerated the digitalisation of their customer and supply chain interactions and internal operations by three to four years. But with this rapid rate of digitalisation, businesses, especially SMEs, are now more susceptible to cybercrimes such as phishing attacks and malware.
In fact, the Asia Pacific region has seen an increased number of cyberattacks in the past year. Check Point research shows a 168% year-on-year increase in the number of cyberattacks in the region in May 2021, with an organisation in Asia Pacific typically suffering 1,245 attacks each week.
With enterprise IT workloads requiring scale and flexibility, Kubernetes is enabling enterprises to deploy a common cloud platform for business applications to be deployed across multiple clouds, delivering more agility, consistency, and reductions in costs and complexity.
There are three fundamental business capabilities that Kubernetes can drive in SMEs:
- Protecting SMEs from Ransomware
With ransomware attacks on the rise and rapidly increasing business stakes, data protection, particularly backup and disaster recovery, is more critical than ever. But ransomware is not only the catalyst. In the same time ransomware attacks have increased, adoption of Kubernetes and containers saw significant acceleration. According to Gartner, 75% of large enterprises in mature economies will have adopted containers by 2024.
These trends have brought us to where we are today with a modern computing paradigm driven by Kubernetes and containers. In Kubernetes deployments, the proper tooling for successful data management might make the difference between a thriving corporation and a complete organisational shutdown. Unfortunately, ransomware attack vectors in Kubernetes applications and environments are numerous and appealing to hackers.
Implementing the necessary procedures to prevent and overcome ransomware attacks is crucial to any business, especially SMEs. Kubernetes is the unifying fabric of modern computing. Using it to mitigate the most pressing data threats in today's risk landscape is one of the best defenses yet.
Furthermore, tools for backup and recovery functions should also promote automation and integrate seamlessly into existing workflows. Enabling immutability, creating backups with unique code paths, protecting backups for maximum effectiveness, and enabling seamless restores are part of a robust ransomware data protection strategy.
- Cut down manual processes
The second fundamental business capability of Kubernetes is its ability to allow businesses to eliminate tedious, time-consuming manual processes and, in turn, freeing up time for the company to focus on growth and innovation.
As Kubernetes and its ecosystem have evolved, automating specific workloads and processes has become much more achievable, such as setting up and installing applications, pod and node scaling, ongoing storage management, and automating security workflows. It also allows you to schedule and run containers on clusters of physical and virtual machines while automating many operational tasks, enabling enterprises to tap into the potential of containers in day-to-day work via an automated fashion.
Kubernetes natively provides a rich set of capabilities to allow traditional and modern workload types to scale dynamically, based on a consistent and declarative deployment model. This allows application developers and operators to define a profile for each workload that's equipped to scale based on demand and recover automatically, ensuring the integrity of the application and guaranteeing the highest levels of performance and reliability.
Overall, Kubernetes can help businesses cut infrastructure costs. It makes a container-based architecture feasible across a portfolio of large enterprise apps, which means businesses can pack together apps using minimal resources to get the most out of their cloud and hardware investments. Meanwhile, apps that need to expand can be placed across pods where they have room to grow.
- Enable Agility Across the IT Ecosystem
As our world becomes increasingly more connected, organisations must consider how business agility can be extended to capitalise on the opportunity to deploy applications and services closer to customer needs.
SMEs need to empower their teams of developers, platform operators, and security professionals with a framework that unites many technological platforms spanning private cloud, public cloud, and edge-based computing environments to create an environment of sustained agility.
Kubernetes is evolving to extend the same consistent API that many enterprises use today in their data centres or public cloud settings to new distributed and hyper-connected computing environments. This enables enterprises to extend their applications in a unified manner, leveraging the tech foundation Kubernetes as created.
Given the rising need for better ransomware practices, streamlined business processes, and business agility, SMEs and their IT personnel are increasingly favouring Kubernetes, which provides a plethora of time-saving, productive options for their data storage and management.
Anthony Spiteri is Senior Global Technologist at Veeam Software


title: Kansas family hoping for miracle for 5 year old with inoperable cancer

text:
OLATHE, Kan. — A rare form of pediatric cancer diagnosed in fewer than 300 children each year in the United States is impacting families in the Kansas City metro.
On Oct. 30, Todd and Stacey Sands of Olathe found out their 5-year-old son, Hudson, has diffuse intrinsic pontine glioma, or DIPG, an incurable and inoperable form of cancer.
"The reason they consider it inoperable is because it's intertwined with his brain stem. There's too many good and bad cells intertwined to get rid of the tumor operatively," Stacey said.
For weeks, Hudson had been experiencing headaches. His parents took him to Children's Mercy Hospital, where he was later diagnosed with Stage IV DIPG, the most aggressive form of the disease.
"We were told that his prognosis, it was a year at best," Stacey said.
According to St. Jude Children's Research Center, the survival rate for children with the disease remains very low. On average, the life expectancy from diagnosis is eight to 11 months.
The Michael Mosier Defeat DIPG Foundation says medical advances have greatly improved the survival rates for children diagnosed with most types of cancer, but not for DIPG.
"Only 4% of all of cancer research is spent on pediatric cancers, and a small portion of that percent is spent on brain tumors," Todd said.
Less than a month after his diagnosis, Hudson has already undergone two surgeries. The Sands family said they have spent countless hours researching their options.
"It's incredible the amount of research and effort that is placed on the parents,” Stacey said. “We got little to no direction on these clinical trials from the doctors that we've spoken with.”
With fewer than five clinical trials available, they are hopeful Hudson is eligible for one. Stacey said doctors will not tell the family which ones Hudson should try, because none have been proven to work.
The family plans to leave for Houston, Texas, to see if Hudson qualifies for the ONC201 trial.
"We're narrowing in on this ONC201 trial that is out there because it's the only thing that we can see that may give us a shred of hope," Stacey said.
If selected for the trial, Hudson would undergo daily radiation and medication for the next two months. His mother says there's an 80% chance the radiation will shrink the tumor, but over time, it could grow back even larger.
“It is likely that within a matter of weeks or months, it will come back, grow large and he won’t survive. But that’s why we are doing everything we can and praying for a miracle," Stacey said.
After the clinical trial, the Sands family plans to take Hudson to all of his favorite places.
A GoFundMe page was created for Hudson, with donations to go toward medical and travel expenses. So far, it has raised nearly $50,000 of its $100,000 goal. Any unused money will be donated to pediatric brain cancer research.
This story was originally published by McKenzie Nelson at KSHB.


title: Boost your business with dg zhen ren shi xun yg88 net

text:
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title: Sap young professionals program

text:
Get to know us and learn how Dubai Internet City supports tech businesses.
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title: Calories in baked pork chop

text:
Pork chop is equivalent to a beef steak and is an expensive portion of the animal. All kinds of pork chops are extracted from the animal’s loin area that extends from its hip to shoulders. The commonly seen chops in butcher shops are extracted from the animal’s loin and ribs. Other chop varieties include sirloin and shoulder chops. However, do you know how much calories are there in baked pork chop? Do read on.
Calories in Pork Chop
Do You Know Four Different Kinds of Pork Chop?
It is also known as shoulder chop, pork loin, pork shoulder steak, pork steak and blade steak. It comes from the animal’s shoulder region and it is a dark colored meat containing a lot of fat, connective tissue and some quantities of blade bones.
It is also known as sirloin steak. It is a cheap cut that comes from the hip region of the animal’s back loin. It contains backbone and hip and has a higher quantity of bone compared to that found in other chop varieties. Its meat comprises of different muscle groups. It has a good pork flavor.
It is also known as rib end-cut, rib pork chop and center-cut rib chop. It comes from the animal’s rib in the loin that runs from shoulder to the loin’s middle. It is a large portion of the lean loin region and doesn’t contain any tenderloin meat. A bone runs along on a side and at times, there’s a fat layer on its outside. More connective tissue and fat is found on rib chops cut off from the end of the blade. These chops are quite soft, have mild flavor and more fat compared to loin chops.
Also known as porterhouse and top-loin chop, a loin chop comes from the loin and hip portion from the animal's back. It may or may not have tenderloin pieces. It contains T shaped bone with one side containing tenderloin and the other loin. It is lean and has moderate pork flavor.
Low Calories and Healthy Pork Chop Recipe
Here is a nice recipe for you that has low calories in baked pork chop. Here are ingredients you will need.
Mix first three ingredients together and sprinkle over pork chops.
Heat nonstick skillet on medium to high heat and coat it using cooking spray. Add in pork chops and cook for 4-5 minutes on both sides or till it is done.
Mix the other ingredients and serve them with chops.


title: Gower

text:
Just a short drive from Swansea, the Gower Peninsula is more than just a pretty face. Now known as Gower National Landscape – and area of outstanding natural beauty, it was designated as the UK’s first Area of Outstanding Natural Beauty back in 1956.
The Gower Peninsula is a rich and varied environment which is as ecologically diverse as it is beautiful – from open, wild moors and dramatic limestone cliffs to golden, sandy beaches. Gower is a geological goldmine with inspiring landscapes and a captivating coastline.
Rhossili Bay has been voted into the top ten of beaches in Britain on numerous occasions, the sunsets are a must-see! Three Cliffs Bay’s iconic vista provides a photo stop for walkers on the Gower coast path. Cefn Bryn common is home to neolithic burial chamber King Arthur’s Stone and the intrigue continues with the peculiar cliff structure Culver Hole near Port Eynon.
With some of the most awesome waves in Britain, sandy Llangennith has been popular amongst surfers for decades. True adrenaline junkies can try a spot of coasteering – climb the cliffs and leap into the waves below, not for the faint hearted! Make sure you’re always accompanied by a professional– there are a number of activity providers who would be delighted to hear from you. And if that sounds a bit too much, you can always hop aboard boat and do a bit of seal and dolphin spotting from the sea.
Find out why Gower has a special place in so many hearts – check out Gower’s breath taking beaches, see what all the fuss is about and make a plan to visit us!
Coast and countryside
The 19 mile-long Peninsula starts at Mumbles and extends westwards. It’s famous for its jaw-dropping coastline and beaches (from vast Rhossili Bay to cosy and secluded Pwll Du), and is a favourite destination for walkers and surfers. Inland you’ll find sheltered woodland and rolling grasslands; country pubs and fine food.
It’s no surprise that walking in Gower is so popular, its home to some of the most beautiful sections of the Wales Coast Path (even though we are biased). Grab your boots and be prepared for a spectacular waterfront walk. It’s not all about sea and sand either – you’ll encounter lush woodlands and stunning countryside views along the way
Sample some of the local delicacies, such as Penclawdd Cockles from the Loughor estuary, succulent Salt Marsh Lamb and the famous Laverbread (its seaweed!).
Outstanding Natural Beauty
Covering 188 sq km, Gower was awarded its AONB status for its classic coastline (much of it is Heritage Coast) and its outstanding natural environment (33% is National Nature Reserve or a Site of Special Scientific Interest).
The Peninsula’s richly varied natural environment is renowned for its heathland, limestone grassland, fresh – and salt-water marshes, dunes and oak woodlands. Its mixed geology has given rise to a wide variety of scenery in a relatively small landscape area. Dramatic limestone cliffs, interspersed with sandy beaches and rocky shores, dominate its southern coast. In the north, the coast is low-lying with extensive salt marshes and dune systems.
Travel back in time
There are at least 1200 archaeological sites on Gower of different periods and types. These include caves, Iron Age forts, medieval castles, churches, a lighthouse and 19th Century parks. 73 of these are of national importance, with 124 listed buildings. The western part of Gower is included in the Register of Landscapes of Outstanding Historic Interest in Wales, for the rich evidence of a long sequence of land use and occupation from the prehistoric to industrial periods. This includes Iron Age forts and a surviving medieval open field system (known as the Vile, near Rhossili).


title: Osondu c nwokoro

text:
The Management is coordinated by Osondu C. Nwokoro who is a communications law, policy and strategy specialist and has been very actively involved in the development of the policy, legal and regulatory frameworks for the communications industry since the very early stages of its partial liberalization in 1996 through the attainment of full liberalization, up till the present. He had worked at the Nigerian Communications Commission where he was deeply involved in the formulation of the pioneering industry regulatory policies and guidelines and also, in the private sector as a long serving Regulatory Affairs Executive at Airtel Networks Limited. During these periods he served as a member of the Telecommunications Sector Reform Implementation Committee of the National Council on Privatization that wrote the National Telecommunications Policy of 2000 (NTP) and was also very actively involved in the drafting of the Nigeria Communications Act, 2003 which is the governing statutory instrument for the communications industry in Nigeria. He was also chairman of the regulatory sub-group of the National Committee on the review of the NTP, a member of the Presidential Committee on the National Broadband Plan (2020-2025) as well as the Expert Review Committee on the National Digital Economy Policy and Strategy, 2019. These experiences have afforded Osondu deep passion for and understanding of the issues around digital access in Nigeria.
Osondu is presently founding managing counsel of Greenfields Law, a Nigerian law firm with specialty in communications and technology, public policy and government affairs, stakeholder management and regulatory compliance. In these contexts, Greenfields Law undertakes assignments around policy and regulatory interventions for digital inclusion and has provided Technical Assistance to the UK Government Prosperity Fund – Nigeria on fostering Effective Right of Way (RoW) Administration and Issuance of Planning Permits for Masts and Towers in Nigeria, a landmark Study which presents pragmatic solutions to the Nigerian government for resolving the long-drawn-out issues around disparate Right of Way and site build permitting processes which militate against broadband access and digital inclusion. In addition to subsequently advising the Anambra State Government (ANSG) on the amendment of its Planning Law in the above respect, the firm has also recently coordinated another Study commissioned by the Foreign, Commonwealth and Development Office on digital access for persons living with disabilities and in underserved locations during the COVID-19 pandemic with reference to the adequacy of the governing policy, legal and regulatory prescriptions among other considerations.
Osondu is an independent adviser to the boards of a number of Nigerian agritech and IT hub companies and has been inducted as one of the One Hundred Leading Telecommunications and ICT Personalities in Nigeria by the Association of Telecommunications Companies of Nigeria (ATCON).


title: 3926

text:
One major hallmark of Alzheimer's disease (AD) is the massive loss of synapses that occurs at an early clinical stage of the disease. In this study, we characterize alterations in spine density and the expression of synapse-associated immediate early gene Arc (activity-regulated cytoskeleton-associated protein) in the hippocampal CA1 regions of two different amyloid precursor protein (APP) transgenic mouse lines before plaque development and their connection to performance in hippocampus-dependent memory tests. The density of mushroom-type spines was reduced by 34% in the basal dendrites proximal to the soma of CA1 pyramidal neurons in 5.5-month-old Tg2576 mice, carrying the Swedish mutation, compared with wild-type littermates. A similar reduction of 42% was confirmed in the same region of 8-month-old APP/Lo mice, carrying the London mutation. In this strain, the reduction extended to the distal dendritic spines (28%), although no differences were found in apical dendrites in either transgenic mouse line. Both transgenic mice lines presented a significant increase in Arc protein expression in CA1 compared with controls, suggesting rather an overactivity and increased spine turnover that was supported by a significant decrease in number of somatostatin-immunopositive inhibitory interneurons in the stratum oriens of CA1. Behaviorally, the transgenic mice showed decrease freezing in the fear contextual conditioning test and impairment in spatial memory assessed by Morris water maze test. These data indicate that cognitive impairment in APP transgenic mice is correlated with impairment of synaptic connectivity in hippocampal CA1, probably attributable to loss of inhibitory interneurons and subsequent hyperactivity.
Alzheimer's disease (AD) is an age-dependent neurodegenerative disorder that is characterized by a progressive cognitive decline and by several structural brain alterations, such as deposition of amyloid β (Aβ) peptides in parenchymal plaques and cerebral blood vessels, intraneuronal formation of neurofibrillar tangles, and loss of neuronal subpopulations (Terry et al., 1998). One major hallmark of AD is the massive loss of synapses that occurs at an early clinical stage of the disease (Masliah et al., 1991; Sze et al., 1997). Synaptic loss has been correlated with cognitive deficits in AD patients (DeKosky and Scheff, 1990; Terry et al., 1991; Lippa et al., 1992). Although a detailed morphological analysis of changes in synaptic connectivity in the AD brain over time is difficult and limited to the use of postmortem material, the generation of amyloid precursor protein (APP) transgenic (Tg) mice that overproduce Aβ has enabled a better understanding of the functional and morphological consequences of Aβ overproduction.
Dendritic spines are anatomical specializations on neuronal cells forming distinct compartments that isolate input from different synapses and are essential for excitatory synaptic transmission (Bonhoeffer and Yuste, 2002). As postsynaptic dendritic contacts, decreases in spine numbers might correlate with the loss of afferent synaptic terminals (Parnavelas et al., 1974; Gould et al., 1990; Cheng et al., 1997). Several in vitro studies have demonstrated that oligomeric aggregates of Aβ cause a reduction in the number and/or length of dendritic spines in hippocampal neurons (Calabrese et al., 2007; Lacor et al., 2007; Shankar et al., 2007). However, a similar reduction of dendritic spine density in the CA1 region of the hippocampus by in vivo-generated Aβ oligomers, e.g., in young APP transgenic animals, has not been shown. The aim of this study was to evaluate the impact of Aβ overexpression on the number of dendritic spines of CA1 pyramidal neurons in two transgenic APP mouse lines, before onset of plaque formation. The Swedish (Tg2576) and London (APP/Lo) mutations are recognized APP transgenic mouse models for preclinical and drug developmental studies (Hsiao et al., 1996; Moechars et al., 1999; Tanghe et al., 2010).
Although the spine density of pyramidal neurons was determined by the Golgi–Cox method (Gibb and Kolb, 1998), accompanying pathological effects were analyzed by staining the immediate-early gene product Arc (activity-regulated cytoskeleton-associated protein), also named Arg3.1 (activity-regulated gene of 3.1 kb) as a marker for synaptic functional and morphological changes, somatostatin (SOM) immunohistochemistry as a marker of inhibitory interneuron integrity, and memory tests as a marker of functional consequences of synaptic changes in same individuals. This is the first study showing morphological alterations in spine density in specific dendritic regions of CA1 pyramidal neurons with possible consequences on spine turnover and inhibitory interneurons, and ultimately cognitive behavior, in two different APP transgenic mouse lines before onset of plaque formation. Presence of pathology at early age suggests that soluble oligomeric rather than deposited fibrillar Aβ is responsible for these deficits.
Materials and Methods
Female Tg2576 mice (5.5 months old; Taconic Farms), APP/Lo mice (8 months old; bred in-house), and age-matched littermate wild-type control mice were housed individually in Macrolon type 2 cages under standard laboratory conditions, 21 ± 1°C air temperature, and 55 ± 15% humidity, with access to food and water ad libitum. Experimental procedures were approved by Abbott's Animal Welfare Officer and were performed in accordance with the European and German national guidelines as well as recommendations and policies of the U.S. National Institutes of Health Principles of Laboratory Animal Care (1996 edition). All experiments were performed during the light phase of the 12 h day/night cycle.
In house, we have validated the contextual fear conditioning test with Tg2576 mice, but for technical reasons, in APP/Lo mice, a modified Morris water maze has been proven to be most reliable (Moechars et al., 1999; Tangle et al., 2010). This strain selectivity for deficits in different tasks may be attributable to genetic background. Thus, we have applied a different test to each strain of mice as described below.
Tg2576 mice and contextual fear conditioning.
Mice were placed into the conditioning box (Hamilton Kinder), and the computer program was started. Mice were given 2 min to explore the conditioning chamber and then presented with a tone (70 dB, 15 s). The tone stimulus coterminated with a footshock (0.78 mA, 2 s). This stimuli pairing was repeated two times, with presentations spaced 2 min apart. Thirty seconds after the second shock, the mice were removed from the chamber. Mice were returned to the home cage and colony room. Before the first mouse and between mice, the chambers were wiped clean with isopropyl alcohol and allowed to dry before testing.
Twenty-four hours later, mice were placed back into the conditioning chamber for a 5 min contextual conditioning test without tone or shock presentation, in which the mice were scored for the presence or absence of freezing (lack of movement except for respiration). Mice were removed from the chamber after the 5 min test, returned to the home cage, and moved to another test room for the novel context test. After 1 h of habituation, mice were placed into the novel context box that had several differences from that of the test chamber: the lighting was reduced, chambers were made of clear Plexiglas and were larger in size than the contextual conditioning chambers, and the floor of the chambers were smooth, white plastic that were coated lightly with a 1% vanilla extract solution (reapplied between mice). The computer program consisted of 3 min without any stimulus presentation, followed by 3 min of the tone stimulus. Mice were scored for the presence or absence of freezing in the first 3 min to examine differences in freezing to the novel context and for the 3 min of tone presentation to assess the memory for the tone previously paired with the footshock.
APP/Lo mice and water maze.
A custom-made water pool (100 cm diameter) with white, nontransparent walls was located in a well-lit room with a number of distal extramaze cues. The water (25°C), which was made opaque with coffee creamer, was filled and drained daily up to ∼10 cm from the top of the pool edge. A clear Plexiglas escape platform (9 cm diameter) was located 0.5 cm below the water surface in the center of one of eight equally sized pool sectors (∼20 cm from the edge of the platform to the pool wall). The platform location was selected randomly for each mouse but was kept constant for each individual mouse throughout training. For each trial, a mouse was released from any of eight start boxes (filled with water and separated from the pool by the manually operated sliding door) and had 60 s to locate the escape platform, in which it was allowed to remain for 10 s. The mouse was then returned to its holding cage, which was located under an incandescent red heat lamp. If the platform was not found in the allotted time, the mouse was gently placed on the platform using an appropriately sized plastic spoon. The mice were given two trials per day for 8 d. Four mice were trained successively in a group enabling intertrial intervals to be 5 min. The mice were tracked in the pool by an Ethovision video tracking system (Noldus).
After behavioral testing, animals were deeply anesthetized by intraperitoneal injection with a mixture of 50 mg/ml ketamine and 10 mg/ml xylazine (200 μl/100 g body weight) and with 0.1 mg/ml atropine and perfused transcardially with 0.1 m PBS. Aware of the sensitivity and immediate effects of handling on the expression of Arc and spine formation/elimination, we kept the time interval between behavioral testing and the time when the animals were killed between 60 and 90 min in all animals.
Directly after perfusion, the right hemisphere was dissected out and processed for a modified Golgi–Cox staining as described by the manufacturer (Rapid Golgi; FD NeuroTechnologies). Coronal sections (240 μm) were obtained using a freezing-sliding microtome (Frigomobil 1206; Reichert-Jung), mounted on 2% gelatin-coated glass slides, and stored in Golgi solution in the dark at room temperature. After 2 weeks, sections were rinsed, dehydrated, cleared of xylene, and mounted onto gelatinized glass slides. Slides were coverslipped and allowed to dry before quantitative analysis. Several pyramidal neurons impregnated with the Golgi solution were readily identified in the dorsal hippocampal region by their characteristic triangular soma shape and numerous dendritic spines (Robinson and Kolb, 1997). At least four neurons per animal were three-dimensionally reconstructed by NeuroLucida Software (MicroBrightField). For spine quantification, a 100× oil-immersion objective was used to identify mushroom-like spines in dendrites longer than 10 μm. Straight branches were preferred to have a clear presentation of mushroom-like spines. At least three proximal (30–120 μm from soma) and three distal (220–340 μm from soma) dendritic segments were quantified in each neuron (Fig. 1). Spine densities were calculated as mean numbers of spines per micrometer per dendrite per neuron in individual mice per group. Spines located on proximal dendrites and spines located on distal dendrites were analyzed separately.
After brain removal, the left hemisphere was immersion fixed in 4% formaldehyde in PBS for ∼24 h at 4°C with subsequent transfer in 30% sucrose/phosphate buffer for at least 3 d. Thereafter, eight series of coronal sections were cut at 40 μm on a freezing microtome. Accordingly, sections were stained slide mounted for detection of Arc protein levels or free floating for somatostatin-positive cells quantification. Residual series were kept frozen in cryoprotectant medium for long-term storage.
Activity-regulated cytoskeleton-associated protein (Arc).
An automated staining system was used for Arc immunohistochemistry (Discovery; Roche Diagnostics). Briefly, after incubation for 30 min in 0.25% H2O2/ methanol solution and washing steps with PBS, sections were mounted on SuperFrost Plus slides (Menzel Glaeser), air dried, and subsequently transferred into the automated staining system. For all stainings, the diaminobenzidine (DAB) technique was used for chromogenic reaction (DAB Map kit; Roche Diagnostics). Briefly, after a fixation step with 4% formaldehyde in PBS/reaction buffer for 12 min, followed by a mild CC1 heat pretreatment (delivered from Roche Diagnostics), slides were incubated with primary antisera in antibody diluent (Roche Diagnostics) at room temperature for 6 h. The Arc antibody was used at a dilution of 1:200 (applied concentration, C-7; Santa Cruz Biotechnology). Thereafter, slides were exposed to the matching biotinylated secondary antibody in antibody diluent (Roche Diagnostics) for 30 min at 37°C. The secondary antibody was used at a dilution of 1:500 (applied concentration, donkey anti-mouse IgG; Jackson ImmunoResearch). All other steps (blocking steps, chromogenic DAB reaction, etc.), which are included in the DAB Map kit procedure, are standardized and controlled by the Discovery autostainer and cannot be influenced by the researcher. After finalization of automated staining, slides were washed in normal water, dehydrated in graded ethanols, cleared in XTRA-Solve (J. T. Baker), and coverslipped with UltraKitt (J. T. Baker). Samples with omission of primary antibody were included as negative controls in the same run and confirmed antibody specificity because in no case staining was seen. Furthermore, brain sections from various mice with known expression pattern were used as positive controls.
Free-floating coronal sections (40 μm thick) from Tg2576 and wild-type mice were pretreated with 3% H2O2/methanol in 0.1 m PBS, pH 7.4, for 30 min and with an avidin/biotin blocking kit (Vector Laboratories) for 30 min. Sections were rinsed two times with Tris-buffered saline Tween (TBST) buffer and then blocked for endogenous secondary antibody with 5% normal goat-serum in TBST. Thereafter, sections were incubated with a polyclonal rabbit antibody against somatostatin (Santa Cruz Biotechnology) diluted 1:1000 in 1% normal goat-serum/TBST overnight at room temperature. Sections were rinsed and incubated for 30 min at room temperature in a 1:500 dilution anti-rabbit IgG (Vector Laboratories). Thereafter, sections were incubated with Vectastain ABC Reagent for 30 min (Vector Laboratories) at room temperature, and the DAB technique was used for chromogenic reaction. After finalization of staining, slides were washed in normal water, dehydrated in graded ethanol, cleared in XTRA-Solve (J. T. Baker), and coverslipped with UltraKitt (J. T. Baker). Samples with omission of primary antibody were included as negative controls.
Immunohistochemistry image analysis.
For analysis, we used five to seven labeled sections per animal (40 μm thick, ∼320 μm apart, starting at approximately −2.8 mm posterior to bregma) (Paxinos and Franklin, 2001). For Arc image analysis, sections were digitized to grayscale pictures using a Carl Zeiss Axioplan 2 microscope and analyzed for optical densities using Image-Pro Plus software (Media Cybernetics). For somatostain-positive cell analysis, manual quantification was done by using the Neurolucida System (MicroBrightField). Depending on the dimension of the analyzed region, specific a priori-designed and calibrated areas of interest were used for determination of optical densities or number of positive cells within a given region. CA1 dorsal hippocampal region was identified by anatomical hallmarks and according to standard mouse stereotaxic brain atlas (Paxinos and Franklin, 2001). A square area of ∼1.875 μm2 (for both stratum pyramidale and stratum oriens of CA1) and 22.500 μm2 (stratum radiatum of CA1) was used for analysis of Arc staining. At least two measurements of the optical density were obtained from each region within a given section and averaged. Background optical density values, obtained from coverslipped glass slides without sections, were determined and set to 100% transmission. Additionally, all data of Arc optical density measurements were corrected for tissue background staining by subtracting optical density values from fiber tracts within the same section, e.g., corpus callosum. For somatostatin-positive cell counting, a frame covering the whole surface area of each region of interest (mean values for CA1 stratum oriens, 381,625 μm2; CA1 stratum lacunosum-moleculare, 299,812 μm2; dentate gyrus, 69,539 μm2) (Fig. 1) was marked. Then, the number of somatostatin-positive neurons was counted. An average of five to seven transverse sections (320 μm apart) were measured in each animal. The CA1 dorsal hippocampal region was defined using a 2.5× objective, and the number of neurons was counted using a 40×/1.3 oil differential interference contrast objective following the description made by Vela et al. (2003). Briefly, cell density values were obtained by dividing the number of immunoreactive neurons by the total area of the corresponding hippocampal region of interest (see above). The density of immunolabeled neurons from different hippocampal regions was averaged and expressed as the mean number of positive cells per square millimeter. It was not our intention to determine the total immunostained cell number in the whole mouse hippocampus. We are aware that, if this was the aim, Cavalieri point count in combination with three-dimensional optical dissector methods would be more appropriate (Ramos et al., 2006). All analysis was performed by a scientist blinded to the genotype status of the animal.
All data are presented as mean + SEM, and p < 0.05 was considered statistically significant. Contextual conditioning data was assessed by unpaired t test. For water maze data, measures of latency to find the platform were obtained for each trial and then averaged daily values were subjected to two-way ANOVA with repeated measures on time. Whenever applicable for repeated measures analysis, Mauchly's test of sphericity was applied and the degrees of freedom were corrected to more conservative values using the Huynh–Feldt's ε for any terms involving factors in which the sphericity assumption was violated. Statistical analysis of the spine density was performed with the mean total number of spines per micrometer per neuron per mouse for each group, which were compared by unpaired t test. For optical density (Arc) and cell count (somatostatin) analyses, an unpaired t test was used.
Contextual conditioning was assessed in Tg2576 mice by analyzing the time spent freezing in the conditioning context on the test day (day 2). Tg2576 mice spent significantly less time freezing than the wild-type controls (t = 4.023, df = 16; p < 0.01) (Fig. 2A). When tested in the novel context, wild-type and transgenic mice did not differ in the levels of non-associative freezing (p = 0.92) (Fig. 2A). The performance in the water maze was tested in APP/Lo mice. Over 7 d of training, mice improved their ability to find the submerged platform. During the last day of training, wild-type animals had shorter latencies to find the platform than those in APP/Lo mice (Fig. 2B) (genotype × time interaction, F(1,156) = 10.82, p < 0.001).
Density of synaptic spines in CA1 of the hippocampus
To study the anatomical basis of the cognitive deficit in the hippocampus-based tests, synaptic connections were quantified by counting mushroom-like spines in the dendritic region of hippocampal CA1. A total of 18–20 pyramidal neurons in each wild-type and transgenic mice were included for final analysis (n ≥ 6 animals per group). All of the neurons resided in the dorsal part of the CA1 hippocampal region and were completely stained along basal and apical dendrites. Segregation analysis was done to differentiate basal from apical dendrites in the same neuron (Fig. 1). In Tg2576 mice, the spine density in basal dendrites proximal to soma were reduced by 34% compared with wild-type littermates (t = 4.68, df = 8; p < 0.01). Basal dendrites distal to soma and apical dendrites both proximal and distal to the soma presented similar values between transgenic and control mouse (Fig. 3A).
In APP/Lo mice, the number of spines in basal dendrites proximal to soma were also reduced by 42% compared with wild-type littermates (t = 5.224, df = 9; p < 0.001). In addition, these mice had also 28% less spines in basal dendrites distal to soma (t = 2.877, df = 9; p < 0.05). Again, in apical dendrites, no significant differences were observed in either proximal or distal segments (Fig. 3B).
Immunohistochemical analysis of Arc protein
Based on the reduced spine density in hippocampal CA1, the expression of the neuronal activity-related synaptic protein Arc was studied. The region-specific reduction of synaptic spines suggested also a determination of Arc protein levels in three areas of interest within hippocampal CA1: stratum oriens (corresponding to basal dendrites), stratum pyramidale (soma of pyramidal cells), and stratum radiatum (corresponding to apical dendrites). Based on previous reports (Miyashita et al., 2009; Lonergan et al., 2010), we expected a robust Arc protein response at 60–90 min after behavioral testing. Moreover, we have reported previously an elevated number of Arc immunopositive granule cells in dentate gyrus 2 h after exposure to a novel environment in both wild-type animals and Tg2576 mice (Nolte el al., 2008). To test whether the behavioral test or already the genetic background has an effect on Arc expression in APP transgenic mice, we conducted an experiment in which APP/Lo and wild-type mice were handled for 4 weeks without behavioral test before being killed. We found that the untreated APP/Lo mice presented a significantly higher Arc protein level compared with wild type in stratum oriens, stratum radiatum (both p < 0.001), and stratum pyramidale (p < 0.05), indicating that regardless of environmental stimulation, APP transgenic mice present an increased baseline level of Arc protein expression (Fig. 4).
Interestingly, when animals were submitted to behavioral paradigms (i.e., fear conditioning or water maze), the same effect was observed. After the fear conditioning test, Tg2576 mice had a higher density of Arc immunostaining compared with wild-type littermates in all hippocampal regions (p < 0.0001) (Fig. 5A). In APP/Lo mice submitted to the water maze, Arc protein expression was also enhanced compared with wild-type controls, although less dramatically than in Tg2576 mice. The strongest increase compared with the wild-type littermates was found in stratum radiatum (p < 0.0001) and stratum oriens (p < 0.01), with a smaller but still significant increase in stratum pyramidale (p < 0.05) (Fig. 5B). Therefore, the increased Arc protein expression in APP transgenic mice seems to be attributable to a higher baseline level compared with wild-type controls rather than a stronger response to external stimulation.
Quantification of somatostatin-positive cells in Tg2576 mice
Among others, a possible explanation for an increased Arc staining and reduced spine density in stratum oriens of hippocampal CA1 would be the loss of inhibitory interneurons. Therefore, these interneurons were identified and quantified by somatostatin immunohistochemistry. The focus of analysis was laid on areas with considerable presence of SOM-positive cells. This subpopulation of GABAergic interneurons has a limited distribution in CA1 hippocampal region, in which highest density is found in stratum oriens, but none in stratum radiatum and very few in stratum lacunosum-moleculare (Vela et al., 2003; Ramos et al., 2006). Therefore, we included only stratum oriens and stratum lacunosum-moleculare and the dentate gyrus for comparison into analysis (Fig. 1). The Tg2576 mouse strain was used because it has shown the greatest differences in spine and Arc analysis. In these mice, the density of SOM-positive cells was significantly reduced compared with wild-type controls in stratum oriens (p < 0.05), but no significant differences were observed in stratum lacunosum-moleculare or in dentate gyrus (p > 0.1) (Fig. 6).
In this study, we analyzed several markers of synaptic function in the CA1 hippocampal region known for its vulnerability in AD patients (Van Hoesen et al., 1990; Arendt et al., 1998, 2001). In APP transgenic mice carrying the Swedish or London mutation of familial Alzheimer's disease, learning and memory impairments are detected at young age, even before amyloid plaque deposition, suggesting that overexpression of APP in the brain induces deficits in hippocampus-dependent functions (Hsiao et al., 1996; Dodart et al., 2000).
The role of Aβ in the temporal course of AD and its deposition in the brain in amyloid plaques has been well established (Tanzi and Bertram, 2005). Small soluble Aβ peptide aggregates (oligomers), which are formed before Aβ fibrils, have been linked recently to synaptic alterations in several in vitro and in vivo studies (Lambert et al., 1998; Walsh et al., 2002; Ji et al., 2003). To clearly separate effects of oligomeric Aβ from that of fibrillar deposited Aβ, we studied animals at an age when they are still free of plaques but have high levels of Aβ oligomers. When we quantified spine density in pyramidal neurons of CA1 and compared these data with Arc protein expression and SOM interneurons in the same hippocampal region, a complex pattern of changes was observed. A region-specific reduction of synaptic spines in basal dendrites proximal to the soma, accompanied by a reduction of SOM neurons and increased Arc expression, suggests that different inputs cause differential morphological rearrangements in pyramidal neurons of transgenic mice. These regional morphological rearrangements in spine density in two different transgenic mouse lines suggest a similar soluble Aβ-mediated mechanism of action at least at early stages of the pathology. Moreover, the loss of spines in basal dendrites in Tg2576 and APP/Lo mice correlated with behavioral impairments assessed in the same individuals. Jacobsen et al. (2006) reported a similar correlation of loss of spine number in dentate gyrus and deficit in contextual fear conditioning in 4-month-old Tg2576 mice. There are only few other reports describing spine density changes in the hippocampus of APP transgenic mice. Lanz et al. (2003) compared reduction of synaptic spines at different ages in PDAPP and Tg2576 mice lines. Contrary to our findings, these authors reported decreased number of spines along apical dendrites of pyramidal neurons in CA1 (Lanz et al., 2003). Differences in genetic background and gene inclusions may explain the different results. To date, no other reports have focused in CA1 pyramidal neurons from young AD transgenic mice models.
Why is the spine density in basal but not apical dendrites of hippocampal CA1 reduced in both APP transgenic mouse lines? There are several possible explanations for such selective loss of spines in young APP transgenic mice. First, soluble Aβ is generated at higher concentration in stratum oriens compared with stratum radiatum and stratum lacunosum-moleculare (Buxbaum et al., 1998; Su and Ni, 1998; Lazarov et al., 2002; Reilly et al., 2003) Second, a general Aβ overproduction may target specific cell types especially relevant for local network rearrangement, i.e., GABAergic interneurons. On the apical side, CA1 pyramidal neurons receive synaptic input from Schaffer collaterals proximal to the soma and from the entorhinal cortex (via the perforant path) on distal dendrites, whereas the basal dendrites receive most synaptic input from Schaffer collaterals (Spruston and McBain, 2007). Therefore, synaptic input from the same region (i.e., Schaffer collaterals from CA3) impinges on basal and apical proximal dendrites, but spine loss was only detected on basal dendrites. Therefore, axonal innervation from a different region is probably not the reason for the selective loss of spines in CA1 basal dendrites. Selective rearrangement of spine formation or selective elimination of spines on basal and apical dendrites has been reported previously by other authors. For instance, spatial training in a complex environment affects spines in CA1 pyramidal neurons by increasing its number on basal dendrites but not on apical dendrites (Moser et al., 1997). Normal aging resulted in decrease of the spine density on basal but not apical dendrites in C57BL/6 mice, one of the background strains of the APP transgenic mice of this study (van Bohlen et al., 2006). Reelin may play a role in this selectivity because reelin-insufficient heterozygous mice presented a spine reduction on basal dendrites of CA1 pyramidal neurons compared with wild-type littermates, but no significant changes were found in apical dendritic trees (Liu et al., 2001). Chemical treatment can also region-specifically induce spine reductions in CA1 pyramidal neurons of rodents. Neonatal rats exposed repetitively to low doses of paroxon (an organophosphate-type cholinesterase inhibitor) loose dendritic spine selectively in basal dendrites with no changes in apical dendrites of pyramidal neurons in CA1. These changes were accompanied by a reduction in cholinesterase activity in the whole hippocampus (Santos et al., 2004). Because more cholinergic input arrives in the stratum oriens in which the basal dendrites are located, the authors proposed that inhibition of acetylcholine esterase by paroxon leads to accumulation of acetylcholine in this region, thereby increasing the risk of toxic effects in basal versus apical dendrites (Santos et al., 2004). Moreover, these effects could be mediated by muscarinic receptor activation on GABAergic interneurons. Alterations in local GABAergic cells by this toxic compound were also hypothesized (Santos et al., 2004).
Conversely, our Arc data indicate rather a more general synaptic activation in both APP transgenic mice lines compared with their wild-type littermates. Arc is a well known neuronal marker for neuronal activity whose expression is induced during synaptic activity, and its mRNA is rapidly transported to dendritic processes for local translation (Tzingounis et al., 2006; Bramham et al., 2008). In the present study, we first tested whether baseline levels of Arc are different between wild-type and APP transgenic mice and then whether behavioral exposure will affect those levels. In APP/Lo mice that were not submitted to any behavioral test, Arc protein levels were significantly elevated compared with their wild-type littermates. Moreover, when mice were subjected to a hippocampus-dependent memory test shortly before they were killed, there was a similar pattern of enhanced Arc protein expression. Thus, APP transgenic mice seem to have a higher baseline level of Arc protein expression compared with their wild-type controls, indicating general neuronal overexcitation already at baseline condition. Conversely, increased Arc protein levels along all dendritic regions of CA1 (stratum oriens and stratum radiatum) may imply that synaptic spine reduction is not directly correlated with Arc protein levels. It is important to note that our analysis focused on mushroom-type spines, the most permanent spines with a slower turnover rate compared with filopodia (i.e., a lifespan of 7 d) (Yuste and Bonhoeffer, 2004). Therefore, rapid neuronal activation as detected by Arc immunostaining is probably quite distinct from the more permanent change of the number of mushroom-type spines. This leads to the second hypothesis, in which rearrangements in a whole local network might account for the selective spine loss found in basal proximal dendrites of CA1 pyramidal neurons. The most likely candidate for such a local network is that of the inhibitory interneurons. Arai et al. (1994) described significant changes during single theta bursts in CA1 basal and apical dendritic trees, and the basal dendrites undergo greater long-term potentiation than the apical dendrites (Abraham and Wickens, 1991; Leung et al., 1992). These authors suggest that such variations to be related not only to the more complex architecture in basal dendrites versus apical dendrites but also to the distribution of GABAergic interneurons in specific regions of the hippocampus (Jinno and Kosaka, 2000; Cope et al., 2002; Pawelzik et al., 2002). The idea of a local network dysfunction in the hippocampus of AD patients has been repeatedly suggested (for review, see Burgos-Ramos et al., 2008). Somatostatin, calretinin, and parvalbumin are all markers of GABAergic interneurons in the hippocampus, and they have been used to demonstrate impairment in aged rats by a reduced number of labeled neurons (Vela et al., 2003; Gavilan et al., 2007). Moreover, selective decreases in somatostatin-positive neurons were found in APP×PS1 mice (an AD model with enhanced pathology) at 6 months of age compared with wild-type mice but only in stratum oriens, corresponding to the region in which basal dendrites are located in our analysis (Ramos et al., 2006). Our analysis of somatostatin-positive cell density in Tg2576 mice, i.e., their selective reduction in stratum oriens, confirms and extends these previous reports.
In summary, we suggest that the observed reduction in dendritic spines on basal proximal dendrites is attributable to a loss of GABAergic interneuron afferents, likely causing glutamate and calcium-mediated overexcitation and local toxicity in stratum oriens. Targeting the basal dendritic region of the hippocampal CA1 and specifically protecting its interneuronal population might be a promising therapeutic strategy to overcome cognitive deficits and reduce side effects by more general therapies in AD patients.
This work was supported by the Deutscher Akademischer Austausch Dienst and Consejo Nacional de Ciencia y Technología (C.P.-C. and F.K.). We are grateful to B. Janson, A. Stanek, B. Theobald, and C. Schuster for their excellent help in tissue processing, (immuno-)histochemical stainings, and microscopical analyses.
- Correspondence should be addressed to Dr. Claudia Perez-Cruz, Neuroscience Research, Global Pharmaceutical Research and Development, Abbott, Knollstrasse, D-67061 Ludwigshafen, Germany. and