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About aus vs ind
The following is a special story about aus vs ind, which attracted a lot of discussion and attention from netizens in the United States on 20 20, 2023..
aus vs ind 背景
Australia were at their dominant best, beat India in all facets: Zaheer ... - Cricbuzz India beat Australia India won by an innings and 132 runs - Australia ... - ESPNcricinfo IND vs AUS: 'We Are Very Much Used to it Now,' Says Rohit Sharma on Jasprit Bumrah's Absence - News18 IND vs AUS 3rd Test: Date, Time, Tickets Price, How to Book Tickets Online - The Quint What India, Australia, Sri Lanka need to qualify for the World Test Championship final - Wisden India consider 'brutal' call to dump struggling star and unleash 'red hot' sensation against Aussies - Fox Sports Match Preview - Australia vs India, Australia in India 2022/23, 2nd Test - ESPNcricinfo Third Test between Australia and India moved from Himalayan venue - The Guardian IND vs AUS: 'We are very lucky to have Rohit take over leadership from someone like Virat' says Dravid - The Indian Express IND vs AUS 1st Test Live Streaming Details: When and where to watch Border-Gavaskar Trophy live streaming in India? - The Indian Express Australia vs India: Last 10 test series records, stats, and results - Business Insider India Ind vs Aus Tests | Hayden offers to help solve Aussie batting woes against Indian spinners - The Hindu Recent Match Report - Australia vs India 4th Test 2022/23 - ESPNcricinfo IND vs AUS | Twitter reacts as Australia bulldoze India by 10 wickets to level series - SportsCafe Recent Match Report - Australia vs India 1st ODI 2022/23 - ESPNcricinfo India vs Australia highlights: India 163 all out; Australia need 76 runs to win 3rd Test - Times of India Women's T20 World Cup live stream: How to watch Australia vs. India match on TV and live stream - DraftKings Nation Head defies doubters AGAIN with latest feat as draw confirms Aussies, India WTC final - Fox Sports IND vs AUS: Revealed: Why Glenn Maxwell Is Not A Part Of ... - Cricket Addictor IND Vs AUS, 1st ODI: MCA Invites Superstar Rajinikanth To Watch Match At Wankhede Stadium - ABP Live India vs Australia, 2nd ODI Highlights: AUS thrash IND to level series - Hindustan Times IND vs AUS: 'We changed the way we bowled' Kasprowicz looks back at Australia's last Test triumph in India - The Indian Express IND vs AUS: "Virat Kohli Isn't Comfortable Against ..." : Former ... - Cricket Addictor AUS-W vs IND-W Live Score - ICC Womens T20 World Cup 2023 ... - Sportskeeda AUS vs IND ODI online tickets booking: Check process to book India vs Australia ODI match tickets online - WION Tour game not needed before India series, insists Australia head coach - The Indian Express IND vs AUS: 'Thanks for all the great battles' – Australia team gifts Cheteshwar Pujara a special jersey on his 100th Test - The Indian Express India vs Australia: ICC Announces Ratings For Nagpur, Delhi Pitches - NDTV Sports Steven Smith to captain Australia in ODI series against India - ESPNcricinfo Ganguly dishes out savage IND-AUS outcome even before reporter finishes question - Hindustan Times India vs Australia: My plan hasn't changed for 13 years, says Mitchell Starc after taking 5-wicket haul - India Today Recent Match Report - Australia vs India 2nd Test 2022/23 - ESPNcricinfo Australia and India go toe-to-toe in thrilling semi-final | Women's ... - ICC Cricket India's continuing dominance vs Australia in Test series: The big stats - Indiatimes.com IND vs AUS: R Ashwin reveals the conversation he had with Virat Kohli after Indore Test - The Indian Express India vs Australia: Todd Murphy's family travelled halfway around world to watch him make debut; five-for was worth jet lag, says father - The Indian Express IND vs AUS: I think the right decision was made, says Nathan Lyon on Virat Kohli's controversial dismissal - The Indian Express IND vs AUS series: India vs Australia test & ODI series schedule, venue, time, squad & live streaming details - WION EXPLAINED: How Can Team India Qualify For ICC World Test Championship Final After IND VS AUS 3rd Test? Chec... - Zee News 2nd Test: Ravindra Jadeja takes 10 as India beat Australia by 6 wickets to retain Border-Gavaskar Trophy - Times of India Lack of lower-order runs compound Australia's batting woes - ESPNcricinfo AUS vs IND Commentary - India vs Australia 2023 Updates ... - Sportskeeda Match Preview - Australia vs India, Australia in India 2022, 2nd T20I - ESPNcricinfo IND vs AUS: Swept away in 90 minutes: Harrowed Australia capitulate to the guile of Jadeja, Ashwin; India win by 6 wickets - The Indian Express Match Preview - India vs Australia, Australia in India 2022/23, 3rd Test - ESPNcricinfo India vs Australia: Who has won the most Border-Gavaskar trophies ... - Sporting News How hosts 'lost the spirit of Test cricket' as desperate bid for glory backfires — India View - Fox Sports Highlights, IND vs AUS 2nd ODI 2023: Australia defeat India by 10 wickets - Times of India IND vs AUS: WATCH - India In Deep Trouble As Shubman Gill ... - Cricket Addictor IND vs AUS: Watch- Teammates And Fans Stunned As Ravindra ... - Cricket Addictor India vs Australia: What time does play start for first Test in Australia? - Sporting News 'What a joke': Cricket world blasts farce - news.com.au India vs Australia: Withdrawals a blessing in disguise for Australia? - Indiatimes.com India vs Australia 1st Test: Result, analysis and talking points - Sporting News IND vs AUS: SKY opens up chase, Virat Kohli closes it - The Indian Express Even Steve Waugh's Australia, Clive Lloyd's West Indies can't match India's 15-series winning streak at home - The Indian Express Ashwin-Jadeja: Yin and Yang, AC-DC, a deadly pair that cannot be unhyphenated - The Indian Express Live scorecard: India vs Australia, third Test from Holkar Stadium in Indore - ABC News India vs Australia Test match time, live stream, TV channel, lineups ... - Sporting News Aus vs Ind 4th Test | Shreyas Iyer suffers back pain again, taken for scans - The Hindu When is the India vs Australia Test cricket series? Details, schedule ... - Sporting News Australia's selection woes for Indore Test: Warner and Hazlewood out, Head to open with Khawaja, Green and Starc available - The Indian Express Live scorecard: India vs Australia second Test at the Arun Jaitley Stadium in Delhi - ABC News Cummins returns home due to family health issue - ESPNcricinfo World Test Championship 2023: India's WTC Final chances hinge on series outcome - Indiatimes.com India get the better of Australia, one flick at a time - ESPNcricinfo Kohli shatters Tendulkar's world record to rewrite history in 2nd Test vs AUS - Hindustan Times IND vs AUS ODI Squad:: No change in Team India for last two Tests - Indiatimes.com IND vs AUS: Virat Kohli steals the show by showcasing India's pride ... - Twelfth Man Times From Test No. 2 to going home: the bizarre handling of Ashton Agar - ESPNcricinfo India vs Australia Warm-Up Match Highlights: India beat Australia by six runs - The Indian Express Ellyse Perry pulls off incredible diving stop in Australia vs India semi ... - Sky Sports India beat Australia India won by 6 wickets (with 1 ball remaining ... - ESPNcricinfo Match Preview - Australia vs India, Australia in India 2022/23, 1st Test - ESPNcricinfo India vs Australia: What time does the second Test start in Australia? - Sporting News WTC 2021-23 points table: India nears final spot, Australia drops PCT with Delhi loss; qualification scenarios - Sportstar IND vs AUS: Gautam Gambhir warns India against complacency, reminds how 'Dravid- Laxman' epic stand outplayed Aussies - The Indian Express Full Scorecard of India vs Australia 2nd ODI 2022/23 - Score Report - ESPNcricinfo India vs Australia, 4th Test, Mar 09, Australia tour of India, 2023 - Cricbuzz 'I've been on 2 tours…': Shastri's jaw-dropping IND vs AUS scoreline prediction - Hindustan Times IND vs AUS: Has David Warner played his last Test in India? - The Indian Express IND vs AUS: Why is David Warner Not Playing Today's India vs ... - India Fantasy Australia beat India Australia won by 9 wickets - India vs Australia ... - ESPNcricinfo 'Our stadiums are pathetic': India vs Australia Test match venue draws harsh criticism; ex-CEA urges makeover - Business Today IND vs AUS: KL Rahul deserves more chances as an opener before an in-form Shubman Gill takes his place - The Indian Express Cricket loses it for 'catch of the century' - news.com.au India beat Australia in second Test to take 2-0 series lead as Ravindra Jadeja plays starring role - Sky Sports Marsh's BRUTAL display, Smith's 'catch of the century' contender as records fall in ruthless Aussie win - Fox Sports India beat Australia India won by 5 wickets (with 61 balls remaining ... - ESPNcricinfo IND vs AUS: Disappointed and shell-shocked: Border, Hayden slam Australia's disastrous batting display - The Indian Express IND vs AUS: "The Guys And The Team Are Very Much Used To It" – Rohit Sharma On Jasprit Bumrah's Unavailability - Cricket Addictor Fans Overwhelmed As India Beat Australia By 6 Wickets In 2nd Test ... - NDTV Sports Steve Smith's brutal comment says it all as India humiliated - Fox Sports Australia levels ODI series with India after Mitch Starc takes five wickets in Visakhapatnam - ABC News India v Australia: third Test, day one – as it happened - The Guardian IND vs AUS: 5 Indian Players Who Can Retire From Tests After The ... - Cricket Addictor India vs Australia: Confirmed squads, predicted lineups for 2023 ... - Sporting News 2nd ODI in Numbers: Multiple records broken as Australia sink India - Indiatimes.com India vs Australia: How misreading the pitch resulted in Australian batsmen making the wrong choice of shot – the sweep and slog sweep - The Indian Express Full Scorecard of Australia vs India 1st ODI 2022/23 - Score Report - ESPNcricinfo


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Individuals, because of less active way of livings, are getting weight continuously, and also there seems to be no option for it. If you are just one of these individuals and also you're having a hard time to reduce weight, after that it is necessary that you recognize the concept of inner temperature equilibrium. This implies that your body is frequently attempting to maintain its inner temperature to melt even more calories than it eats at the mobile level.
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By taking these teas, you improve your total wellness and also wellness by eliminating kept waste from your system. This can cause weight reduction as contaminants are eliminated throughout the body, bring about a decrease in calorie intake. These recipes give an excellent start to your weight loss trip with Alpilean.

subset: commoncrawl


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My ticket number with your support department on this is QMI-110-76512
Why is there no option to schedule updates from the deep freeze console?
We are trying to update the deep freeze client on 15,000+ computers accessed via 46 deep freeze console servers.
We cannot run this update by hand via the console during the day as the upgrade requires a reboot and our teachers, admin and students are using the computers so we want to run a scheduled update over the weekend.
We cannot update the client via a batch file as it will not pull the client install file from a network location, only the local computer that is being updated.
Our only option is to log into each 46 of our deep freeze consoles over the weekend and manually wake up the 15,000+ computers and update the clients?
I find it hard to believe that a piece of 20 year old software requires a manual update.


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The foreigner who intends to seek protection from the Italian state can apply because he is fleeing persecution, torture or war, even if he entered Italy illegally and is without documents.
The applicant must justify in the application the circumstances of persecution or serious harm that motivated his flight.
The agents of this persecution or serious harm can be the State, parties or organizations that control the State or a part of its territory or non-state subjects if the State, or whoever controls it, does not want to provide protection to the victim of persecution or serious harm. .
The application must be submitted to the border police or the police headquarters, which does not necessarily have to be the border police, but may be the one in which the applicant intends to be domiciled.
The Police Headquarters issues a document certifying the request and the date of the appointment for the minutes.
Via Venezia 115, Capriccio (PD), 35010 – REGISTERED OFFICE
Via Montà 29, Padova, 35138 – OPERATIVE OFFICE
+39 389 8745065
Hospitality: email@example.com
Foundation: firstname.lastname@example.com
Work: firstname.lastname@example.com
Pec: email@example.com


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this bothers me for a while, that there is no Track for what you Spend your Crystals. (atleast Royal Crystals because they are Realmoney)
So now i got the new Prime Loot, and why ever, a message appeared in Chat when i open the Cart:
Which says +4000 Royal Crystals.
I was wondering "wut, Prime give crystals?!?!". Unlucky it wasnt. That what just my current Owned Crystals.
But my Crystals just stick around waiting for Skins. So i didnt really keep track of them by myself, so i wasnt even sure if that is the correct amount.
After doing research on my Skins and Ark Pass. The amount was Correct.
But i really wish we can get a History ingame. Atleast something like last 180days.
Steam Purches can do It.
League of Legends can do it. And i dont play much more f2p. But i think most games has some kind of Buy History.
Thank you


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Cooperation project FriVer+ startedOn 01.01.2023 the project "European Peace Treaties of the Pre-Modern Period in Data" started at the IEG, in short "FriVer+", funded by the NFDI consortium "Text+".
This project aims to transform the data, in accordance with the FAIR principles, that is made available on the website Europäische Friedensverträge der Vormoderne online. This website provides information about a total of 1800 European peace treaties that originated between 1450 and 1789. The data, comprising digital images of the peace treaties as well as metadata and, in a more limited number of cases, transcriptions and even formal editions of their text, is currently stored in a relational database. Via queries executed on the website the data can be retrieved from this database, but access to the raw data is less straightforward than it ideally should be nowadays. In order to enhance the discoverability of and access to this data, it will be exported from the database and transformed into XML in accordance with the TEI schema. Widely used by the academic community, the use of this schema as well as the inclusion of the transformed data in a public and freely available repository will ensure that the data can be easily used and re-used by scholars from different disciplinary backgrounds, thereby fostering the study of (early modern) peace treaties.
The project, which is generously funded by the NFDI consortium Text +, runs in 2023.
Picture credits: First extension of the First Confederation of the Rhine, Frankfurt am Main, 31.08.1660 (detail), source: Niedersächsisches Landesarchiv Hannover - Arcinsys Digitalisat-Betrachter (niedersachsen.de), NLA HA Cal. Or. 31 No. 430 (Public Domain).


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European Convention
Competent authority in partner country
Individual European competent authorities
Minimum financial contribution
(As a % of the cost of production)
Finance only co-production permitted?
Yes - minimum of 10%, maximum of 25% (Subject to approval by competent authorities)
Third party producers from other countries permitted?
Non party producers allowed?
Non-party allowed with prior agreement from competent authorities
Where the film is made
Filming may be carried out in any country, subject to the requisite amount of points in Appendix II of the European Convention Up to 30% of the total budget may be spent.


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Phytoremediation / Phytomining Are under threat! Rio Tinto – RBG Kew Workshop (July 2001) AIMS: • Current status of metallophytes? • Conservation of Biodiversity? • Applications and risks? • Access to resources and benefits? ... Microsoft PowerPoint Mining_and_Metallophytes
advantages and disadvantages of phytomining Introduction: purification of copper by electrolysis metal extraction reduction of How copper can be extracted from ores by phytomining phytoextraction and bioleaching using 'rock eating' bacteria!
This page contains PowerPoints for the NEW AQA 2016 GCSE course. The entire triple science course is covered in these slideshows. The separate science or higher tier only content is also clearly labelled, meaning that the slideshows can easily be adapted for double science and lower ability groups as well.
The advantages and disadvantages of vibration jaw crusher. The advantages and disadvantages of vibration jaw crusher. Font Size: Views: 852 . Vibration jaw crusher is the essential mineral crushing a crushing equipment, is widely used in mining, building materials, chemical industry, road.
Jan 18, 2015· For big presentations with Slides I'm sure I'd export a backup PowerPoint and PDF file to host locally, but from what I can tell offline access shouldn't be a problem anymore for Google Drive. ... There are lots of advantages, from always having your files backed up in the cloud to being able to keep working if your main computer dies.
Teaching with PowerPoint PowerPoint, when effectively planned and used, can enhance instruction. People are divided on the effectiveness of this ubiquitous software—some say that PowerPoint is wonderful while others bemoan its pervasiveness. No matter which side you take, there are effective ways to use PowerPoint which can be
BENEFITS DRAWBACKS Calcium Carbonate Add water Heat Add more water filter Add CO2 KEY WORDS: DISPLACEMENT ORE BLAST FURNACE ALLOY SMELTING BIOLEACHING PHYTOMINING ASSESSMENT: Put these metals in their order of reactivity Carbon, Magnesium, Copper, Iron Potassium Less reactive metals are displaced by carbon.
Phytoremediation is an emerging technology that has overshadowed earlier physical and chemical technologies used for soil reclamation, particularly because of its economic, industrial and commercial benefits. However, certain factors influence the fluctuating economics of phytoremediation.
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Physics 5 Forces. This PowerPoint covers the NEW 2016 (AQA) GCSE unit called Forces. It covers: vector and scalar quantities, contact and noncontact forces, weight and mass, weight being proportional to mass, centre of mass, resultant force and how it affects motion, drawing a resultant force using scale drawings, vector diagrams, work done,...
Discuss noneconomic benefits, including those related to regulatory requirements. Benefits Realization List each expected benefit, whether it can be measured, and the proposed method for measurement. If the method for measurement does not currently exist, its' development should be included as a project requirement. Measurable Benefits
Full Answer. Mountaintopremoval mining, for example, allows mining companies to extract a significant amount of coal at a far lower cost, but its destructiveness has led to protections for the environment. Traditional forms of mining can be unsafe for workers. Fires and collapses can be deadly, and the health effects of mining can be devastating.
Copper can also be extracted by phytomining. Plants take up copper from the soil. The plants are then burnt and the ashes are put in sulphuric acid. Scrap iron is added to the copper sulphate to displace it. Iron + copper sulphate Copper + iron sulphate. Copper can also be extracted by bioleaching. Bacteria feed on copper ores to get copper ...
62 Advantages and Disadvantages of PowerPoint in Lectures to Science Students two conformations of a molecular is hard to explain using chalk writing, but it could be easily demonstrated by two figures in PowerPoint. For another example, when explaining physicochemical process "sintering", a series


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Managing client relationships is an essential component of running a successful accounting firm. This is especially true for firms that deal with tax compliances and deadlines, as timely communication and efficient processes can make all the difference in ensuring client satisfaction and meeting regulatory requirements. In this article, we will explore some best practices for managing client relationships and communication for a CA firm with lots of tax compliances and deadlines.
Set Clear Expectations
Setting clear expectations with your clients is essential to managing your relationship with them. This includes setting clear timelines for when you will deliver work, as well as the level of communication you will have with them throughout the engagement. Being transparent about what you can and cannot do is crucial in ensuring clients feel heard and valued.
Communication Is Key
Effective communication is vital in maintaining strong client relationships. This includes being proactive in reaching out to clients and providing regular updates on the status of their work. Additionally, you should be available to address any questions or concerns they may have in a timely manner. Use tools like email, phone, and video conferencing to stay connected and establish an open line of communication with your clients.
Automate Processes
Automation can be a significant asset in managing client relationships and communication for a CA firm with a high volume of tax compliances and deadlines. For example, you can use software to automate the creation and delivery of invoices, tax returns, and other documents. This will help to ensure that your clients receive their work in a timely and efficient manner.
Invest in Technology
Investing in technology is another effective way to improve communication and streamline processes. There are numerous software solutions available that can help you manage client data, automate workflows, and improve collaboration between team members. Utilizing these technologies can help you stay organized, improve efficiency, and ultimately provide a better experience for your clients.
Stay Up-to-Date with Tax Regulations
Staying up-to-date with tax regulations is essential for maintaining client relationships and meeting regulatory requirements. This means staying informed about changes in tax laws and regulations, and making any necessary adjustments to your processes to ensure compliance. You can stay informed through industry publications, webinars, and other educational resources.
Establish Trust
Establishing trust is crucial in managing client relationships. This means being transparent, honest, and ethical in all your dealings with clients. It also means providing high-quality work that meets or exceeds their expectations. You can establish trust by being proactive in addressing any issues or concerns that arise and working collaboratively to find solutions.
Personalize the Experience
Personalizing the experience for your clients can go a long way in building strong relationships. This means taking the time to understand their unique needs and preferences, and tailoring your services accordingly. For example, you may choose to offer different levels of service depending on the complexity of the client's tax situation or provide customized reporting that meets their specific needs.
Provide Exceptional Customer Service
Providing exceptional customer service is essential in managing client relationships. This means going above and beyond to meet the needs of your clients, even if it requires extra effort on your part. For example, you may choose to offer extended hours during tax season or provide additional support to clients who are struggling to meet a deadline.
Use Client Feedback to Improve
Using client feedback to improve your services is an effective way to strengthen client relationships. This means soliciting feedback regularly and using it to make improvements to your processes and services. You can do this through surveys, focus groups, or one-on-one discussions with clients.
Celebrate Successes
Celebrating successes with your clients can help to build strong relationships and foster loyalty. This means recognizing their achievements, such as successfully navigating a tax audit or meeting a critical deadline, and celebrating these successes with them. By demonstrating that
you appreciate and value their business, you can build a deeper connection with your clients and establish a culture of collaboration and partnership.
Be Proactive
Proactivity is critical in managing client relationships, especially for a CA firm with lots of tax compliances and deadlines. This means being proactive in identifying potential issues and addressing them before they become significant problems. For example, you may choose to implement a quarterly review process to identify any areas of concern or provide regular check-ins with clients to ensure that everything is going smoothly.
Develop a System for Managing Deadlines
Deadlines are a critical aspect of tax compliance, and it is essential to have a system in place to manage them effectively. This includes identifying all relevant deadlines and establishing a process for ensuring that they are met. It also means having a backup plan in case of unexpected delays or disruptions.
Foster a Culture of Collaboration
Fostering a culture of collaboration is essential in managing client relationships for a CA firm with lots of tax compliances and deadlines. This means promoting open communication and encouraging teamwork among your staff. It also means recognizing the unique strengths and talents of each team member and leveraging them to deliver exceptional client service.
Provide Regular Training and Professional Development
Providing regular training and professional development opportunities for your staff can help them stay up-to-date with changes in tax laws and regulations. This, in turn, will help you deliver better service to your clients and meet regulatory requirements. It also demonstrates your commitment to your staff's growth and development, which can help to foster loyalty and improve retention.
Embrace Continuous Improvement
Embracing continuous improvement is essential in managing client relationships for a CA firm with lots of tax compliances and deadlines. This means regularly reviewing your processes and services to identify areas for improvement. It also means being open to feedback from clients and staff and using it to make necessary adjustments. By embracing a culture of continuous improvement, you can ensure that you are delivering the best possible service to your clients and staying ahead of the competition.
In conclusion, managing client relationships and communication for a CA firm with lots of tax compliances and deadlines requires a proactive and strategic approach. By setting clear expectations, communicating effectively, automating processes, investing in technology, staying up-to-date with tax regulations, establishing trust, personalizing the experience, providing exceptional customer service, using client feedback to improve, celebrating successes, being proactive, developing a system for managing deadlines, fostering a culture of collaboration, providing regular training and professional development, and embracing continuous improvement, you can build strong client relationships, deliver exceptional service, and grow your business.


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Iranian woman 'beaten' by police for 'improper hijab' dies after coma: State media
Mahsa Amini, a 22-year-old Iranian woman who earlier this week fell into a coma shortly after being arrested and allegedly "beaten" by Iran's morality police for "improper hijab" has died, the semi-official Fars news agency reported on Friday.
Amini had earlier this week traveled from Iran's Kurdistan province to the capital Tehran to visit relatives when she was arrested by Iran's morality police, known as "Gasht-e Ershad," for "improper hijab" – that is, not fully covering her hair.
Iran's police had said Amini "suffered a heart problem" while in detention – an assertion activists reject. They say Amini was beaten by police officers.
"Tehran's police announced that Mahsa Amini 'suddenly suffered from a heart problem' – as if a 22-year-old woman has a heart attack that causes a coma and disfigurement naturally. Iranian media are publishing this nonsense as fact," Mahsa Alimardani, a digital-rights researcher with the human rights organization ARTICLE 19, wrote on Twitter.
Hijab, which was made mandatory for women in Iran shortly after the country's 1979 revolution, is considered a red line for Iran's theocratic rulers. Women who break the strict dress code risk being harassed and arrested by Iran's morality police.
Based on the dress code, women are required to fully cover their hair in public and wear long, loose-fitting clothes.
Amini's story has sparked outrage on social media, both from inside and outside Iran.
"What does the supreme leader [Ali Khamenei], who rightly denounced the US police in the death of George Floyd, say about the Iranian police's treatment of Mahsa Amini?" Mahmoud Sadeghi, a former member of Iran's parliament, wrote on Twitter on Friday.
One person with the Twitter handle @rezahajilou pointed out that Amini's right ear looked bloody in a widely shared image of her lying in a hospital bed.
"Mahsa Amini's ears have bled. Most likely, her skull was broken and her ears were bleeding due to a severe beating. The heart attack story is a lie. We are dealing with a terrible crime," the person tweeted.


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Betty Janiece Presser
July 9, 1942-March 9, 2023
Betty Janiece Presser, 80, of Greenfield passed away on Thursday, March 9, 2023 at Westminster Village North in Indianapolis.
She was born July 9, 1942 in Anderson, Indiana to Harold L. and Lillian L. (Woods) Wiseman.
Betty married Ernest Richard Presser on August 16, 1964 at Ford Street United Methodist Church in Lapel.
She was a 1960 graduate of Lapel High School. She earned her bachelor's degree from Purdue University, graduating in 1964.
She was a home economics teacher at East Peoria Community High School for 20+ years. She was a member of the East Peoria Community High School Home Ec Club. She was an avid reader and enjoyed traveling and spending time with her family and friends. She always had a smile on her face.
She was a member of Mossville United Methodist Church in Peoria, Illinois and Ford Street United Methodist Church in Lapel. She also attended Trinity Park United Methodist Church in Greenfield.
Betty is survived by her husband of 58 years, Ernie Presser; her children, David (Jennifer) Presser of Greenfield and Susan (Keith) Nachtigall of Alexandria, Virginia; a brother, Dennis (Susan) Wiseman of South Carolina; and a grandson, Jacob Presser.
She was preceded in death by her parents.
A funeral service will be held at 11:30 a.m. on Wednesday, March 15, 2023 at Hersberger-Bozell Funeral Home, 1010 North Main Street, Lapel, Indiana 46051 with the Rev. Larry Van Camp officiating. Burial will follow in Brookside Cemetery in Lapel.
Visitation will be from 9:30-11:30 a.m. on Wednesday at the funeral home.
Memorial contributions may be made to the Susan G. Komen Foundation for Breast Cancer Research, P.O. Box 6069, Dept. 95, Indianapolis, Indiana 46206-6069 or to the Harold and Lillian Wiseman Educational Scholarship at Coastal Carolina University, c/o: Financial Aid/Scholarships, P.O. Box 261954, Conway, South Carolina 29528-6054.


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Edgar Wright has been delighting geeks around the World for years with his films. From his first cinematic outing with Shaun of the Dead to the dizzying heights of Scott Pilgrim vs. the World. His career has been awash with movie success. But before Wright made it big in Hollywood he working in the British TV industry. His break came when he worked on Spaced. This off-beat sitcom followed a bunch of 20-something Londoners as they bluffed their way through adulthood. It was here that Edgar Wright formed a strong friendship with Simon Pegg and Nick Frost. These actors went on to collaborate with him in 3 of his movies.
Edgar Wright's journey into Hollywood
Kissing goodbye to his British roots, Edgar Wright has now well and truly made a name for himself as a top Hollywood director. Baby Driver, his latest offering, is due to hit cinemas later this year. This film will see Wright take on a different directorial style, having opted for a crime thriller. He has previously delighted fans with comedies, along with a side serving of science fiction. This release will be a much grittier and serious piece of work. Baby Driver centers on a getaway driver who becomes involved with a dangerous gangster. As things progress this young driver is drawn into a robbery, a mission that shows little promise.
Not sure you can wait to get involved in this action-packed thriller? Well, worry no more because the release date has just been moved up. The film had previously been due for release in August. During the CinemaCon celebrations in Las Vegas, earlier in the week, Sony confirmed the release date would be June 28th. Edgar Wright followed up this news by confirming the release date on Twitter. Certainly a date for the diary of any of Wright's fans. Baby Driver will certainly be one to watch at the summer box office.
Orlando Bloom and David Harbour Share the Gran Turismo movie's first trailer
Well, this movie has certainly come together quickly, just like the automobiles the franchise is centered around. In truth, Gran Turismo adaptation directed by Neil Blomkamp has already wrapped up filming and is in post-production ahead of its global theatrical release on August 11, 2023. Do you want to see a brief teaser? To view the trailer, click above.
Orlando Bloom and David Harbour provide some comments in this 60-second clip, which also includes a few brief film snippets. In essence, this is based on the real-life experiences of Jann Mardenborough, who won the 2011 GT Academy competition and later found success as a racing car driver.
In the few photos that were displayed, the cinematography seemed amazing, therefore it is obvious that this would look stunning on a large screen. During Sony's CES press conference, Blomkamp briefly discussed how he is employing the company's cutting-edge cameras to not only get stunning close-ups from within the car's cockpit but also to imitate some of the game's angles, as shown in the trailer.
Geek Culture
"An imposing, spectacular, supersized movie," according to the review of Avatar: The Way of Water
James Cameron was king of the bigger, better, and more contentious sequel blockbuster before he was crowned King of the World. Avatar: The Way of Water comes near enough to retain that reputation, even if his eagerly anticipated return to Pandora can't rival Aliens or T2 for targeted tanker-weight efficiency. And it certainly knocks the flying fish off of Piranha II.
Will it become a $2 billion club member as Cameron suggests it must? We'll see, but it's undeniably flawed yet full of flavor (to paraphrase Guillermo del Toro(opens in new tab)). "MOVIE-MOVIE" is a sometimes strange, always magnificent sensory hit with a thematic thrust that is pleasingly genuine with an undercurrent of soft feeling. Untangling some of the story lines might require numerous viewings and three more movies. But Cameron is the best person to make the case for going to the movies again and again.
Cameron doesn't spend much time setting the scene because the majority of people have already been to Pandora. The first scene quickly parachutes into Pandora's rainforest, where Jake Sully (Sam Worthington) and Neytiri (Zoe Saldana), who have turned completely Na'vi, are now raising their growing family. They have three biological children: Tuktirey, Neteyam (Jamie Flatters), and Lo'ak (Trinity Bliss). Then there are the adoptees: Spider (Jack Champion), a feral human orphan orphaned by war, and Kiri (Sigourney Weaver), a type of offspring of Grace's avatar (from the original Avatar).
Jake feels that defending his family gives him meaning. The Sully family seeks safety among Pandora's sea clans when the evil Colonel Quaritch (Stephen Lang) reappears in (completely explained) "Recombinant" avatar form seeking "payback" for his demise. Jake hovers firmly over his tearaway brood in this instance. But how long can they evade Quaritch's new blue marines? And isn't knowing how to manage risk an important learning curve?
While Cameron's bare-bones setup showcases his pulp punch as a writer, it also demonstrates his astounding skill as a world-builder. This time, Cameron doesn't give much opportunity to pause and take in Pandora's plant life. The lush jungle suddenly seems inhabited and alive. However, the RDA (Resources Development Administration) has grander plans for Pandora, even though life on Earth is hardly sustainable at this point. Their base of operations is a small metropolis with cutting-edge technology like robotic spider "swarm assemblers" that can create structures in a matter of days. Cameron makes sure you can sense the destruction left behind when humanity arrive on Pandora.
The Way of Water definitely has the WOW factor in terms of CG. Thirteen years later, Avatar's spectacle-cinema upgrade has been surpassed. Hair and skin gleam; flames and dust particles transfix. The great revelation this time is the reef, which is home to the Metkayina clan, just as Avatar took time to introduce viewers to Pandora's funky wonderland. The aquatic realm is vivid, sensual, and tranquil. The sense of weightlessness immersed in the waves reveals a new, sensitive grace in Cameron's direction as the 3D visuals shimmer in time with Simon Franglen's ringing score. He instills respect for the ocean in addition to entranced love because the waves are both seductive and hazardous. And the sensation of anguish is overwhelming when their residents are mistreated.
Cameron expertly balances thematic, narrative, emotional, and character strands while dazzles your eyes. Jake's instinctive need to save his children creates danger as a thematic pattern; from the opening monologue on, Cameron treats the theme like a dorsal fin to cling to through stormy story waters. This is somewhat reminiscent of Finding Nemo.
Casting-wise, Saldana and Kate Winslet (as Ronal, the co-leader of the Metkayina tribe) are a little too much in the background, but Worthington shines as the former Na'vi trainee turned training-on-the-job father. Weaver bridges the age gap between actor and character by touchingly projecting Kiri's feelings of exclusion and sulky eye-rolls through the mo-cap. Dalton, one of the fantastic young actors, gives bonding scenes with the whale-like Tulkun heart when they otherwise may have seemed a bit Free Willy. Additionally, Champion dispatches the Newt-like Spider, whose subplot expands on Cameron's family-related ideas.
Although it's unfortunate that his toxic spiel ("science pukes," etc.) sounds familiar, the returning Lang adds explosive wrath. Cameron doesn't spend any time brushing up on Avatar, but he occasionally uses well-known beats. The Sully clan's water-training reworks Jake's previous Na'vi training, while marine animals like the "ilus" are reimaginings of the "ikrans" from Avatar. When a character moans, "Can't believe I'm tied up again," you wonder if a little editing could have been advised. Cameron even repeats himself a little bit within the movie.
Another minor issue with the plot's stop-start nature is how some characters' difficulties seem to go away for long lengths of time. Cameron, though, harnesses prior career highs into a blast of full-bore, high-stakes extravaganza at the film's climactic point to remind you who's in charge. The Abyss' strange wonder, Aliens' kid danger, Titanic's aquatic horror show, and T2's technology are all there and have been enhanced for tension, action, and emotion. Some loose tale threads leave more questions than answers when the fire is out. However, there are three scheduled follow-ups. Even after three hours and more, Cameron's return leaves you wanting more.
Geek Culture
Here is the second advertisement for The Last of Us on HBO
A brand-new trailer for HBO's live-action take on The Last of Us from PlayStation has just been released.
The program, which will debut on HBO Max on January 15, 2023, and on Sky in the UK the following day, will star Pedro Pascal and Bella Ramsey as its main characters.
The Last of Us is the first TV show produced by PlayStation Productions, a division of Sony Interactive Entertainment created to create film and television adaptations of its own game franchises.
The first game's events will be covered, and there may even be The Last of Us Part 2-related material, according to Craig Mazin, the creator of Chernobyl, and Neil Druckmann, vice president of Naughty Dog.
The show's main protagonists, Joel and Ellie, will be portrayed by Pedro Pascal and Bella Ramsey. Sarah, Joel's daughter, is portrayed by Thandie Newton's daughter Nico Parker (Dumbo), while Tommy, Joel's brother, is portrayed by Gabriel Luna (Agents of Shield).
The Last of Us' debut trailer was released by HBO in September.
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Just two weeks before Christmas!
Throughout the year, I receive inspiration from everyone who supports my writing efforts by reading my books and reading my posts on this website and also on the Vitruvian Man and Tucson Poverello House websites. Your comments, reviews and feedback are invaluable to me. Your words keep me energized, particularly when my batteries run low. As a gesture of my appreciation, I am pleased to offer these two gifts
- The Christmas Story from Gaspar, Another Tale of the Christ as a PDF download and
- The Christmas Story from Gaspar, Another Tale of the Christ as an audio presentation produced by my friend and business associate Alistair McKenzie.
A Special thanks to Alistair for producing this audio AND for writing an original Christmas song "We All Know" featured at the conclusion of the narrative. His song captures the essence of the entire story of Gaspar in a very special way.
You have made my life better. I hope these small tokens of gratitude make your Christmas a bit more meaningful.
Your Grateful Friend,
Gene "E.S." Kraay
The Christmas Story PDF


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In my opinion, the Maroon/Burgundy colorway is one of the best colors to wear on sneakers all year round. For the most part, every time a shoe releases in that colorway, it manages to have pretty good success, which is why I'm pretty sure that this Nike Sock Dart will be one of the best colorways ever released on the model.
Rumored to be dropping this holiday season, the Nike Sock Dart Maroon comes dressed different shades of Maroon cover the marled knit fabric on the forefoot as well as the textile heel and collar. Even the signature midfoot strap comes in tonal fashion to help give the shoe a streamlined look and feel. A White speckled midsole and Black outsole provide the only form of contrast. Thoughts on this new colorway? Let us know in the comments below.
via: Nike Blog
Author's Take
I like it. Big fan of the colorway. If I had to pick a color to pick up on the Sock Dart, it would be these.
The Air Jordan 9 Low Pantone Is Coming To Summer Party Near You
The post First Look At The Nike Sock Dart Maroon appeared first on KicksOnFire.com.
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Independence Anesthesia Services (division of Icon Medical) https://www.ias.health/wp-content/uploads/2021/09/JB-Square-200x200-1.png https://www.ias.health https://www.ias.health true
Locum CRNA - Southern Pennsylvania - 114997
Carlisle, PA 17015 US
2023-03-05 2023-04-20 Type of Job: Temporary Job Number: 114997 Provider Type: Certified Registered Nurse Ane
Job Description
Locum CRNA needed in Southern Pennsylvania
- Start Date: ASAP - ongoing
- Coverage Needs Details: Providers must commit to a min of 3 shifts per week to be considered
- Shift: M-F | 7a-3p | but will consider providers who are open to the traditional CRNA schedules
- Cases: full-service general types of anesthesia | no pediatrics, no trauma, no cardiac | labor and delivery but low census
- Other specialties onsite: general, ob, gyn, gi
- EMR: Epic
- Patient Mix: 95% Adult | 5% Peds
- Time to Credential: 60-90 days
- Never worry about your travel accommodations again! In addition to our in-house travel team, IAS is proud to partner with Christopherson Travel for additional after-hours support.
- All Independence providers will have access to comprehensive benefits including healthcare coverage through the Independent Contractors Benefits Association (ICBA).
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1Biomedical Engineering, School of Engineering and Natural Sciences, Ankara Yildirim Beyazit University, Ankara, Türkiye
2MERLAB Application and Research Center, Ankara Yildirim Beyazit University, Ankara, Türkiye
Associate Editor: C. T. Yavuz
Beilstein J. Nanotechnol. 2023, 14, 280–290. https://doi.org/10.3762/bjnano.14.25
Received 25 Sep 2022, Accepted 13 Feb 2023, Published 28 Feb 2023
Achieving scalable and economic methods for manufacturing ordered structures of nanoparticles is an ongoing challenge. Ordered structures of SiO2 nanoparticles have gained increased attention due to the great potential they offer in filtering, separation, drug delivery, optics, electronics, and catalysis. Biomolecules, such as peptides and proteins, have been demonstrated to be useful in the synthesis and self-assembly of inorganic nanostructures. Herein, we describe a simple Stöber-based method wherein both the synthesis and the self-assembly of SiO2 nanoparticles can be facilitated by a silica-binding peptide (SiBP). We demonstrate that the SiBP acts as a multirole agent when used alone or in combination with a strong base catalyst (NH3). When used alone, SiBP catalyzes the hydrolysis of precursor molecules in a dose-dependent manner and produces 17–20 nm SiO2 particles organized in colloidal gels. When used in combination with NH3, the SiBP produces smaller and more uniformly distributed submicrometer particles. The SiBP also improves the long-range self-assembly of the as-grown particles into an opal-like structure by changing the surface charge, without any need for further modification or processing of the particles. The results presented here provide a biomimetic route to the single-step synthesis and assembly of SiO2 nanoparticles into colloidal gels or opal-like structures.
Keywords: biocatalysis; biomimetics; nanoparticle; peptide; self-assembly; silica
Ordered structures of nanoparticles have gained increased attention due to the great potential they offer in filtering, separation, drug delivery, optics, electronics, and catalysis [1-5]. Nanoparticles with ordered 3D structures, such as supra-particles or super lattices, can possess properties that are not observed in the bulk material or in individual nanoparticles [6,7]. Manufacturing such structures in a well-defined, controllable, and scalable manner is an ongoing challenge. One of the most common strategies to this end are top-down lithographic techniques. While these techniques can yield well-defined and controllable structures, high cost, labor-intensiveness, resolution limits, and high throughput time limit the scalability [8]. Self-assembly allows to circumvent some of the constraints of the top-down techniques to obtain ordered 2D or 3D nanostructures. Self-assembly, however, presents challenges of its own. One major challenge is the difficulty in manipulating nanoparticles due to size-related constraints. The self-assembly of nanoparticles is mainly governed through intermolecular interactions [9]. The high nanoparticle/volume fractions required for large-scale applications may result in electrostatic repulsion or molecular crowding-like effects, preventing efficient assembly of the particles. Therefore, tailoring intermolecular interactions between nanoparticles by modifying the particle surfaces or through external influences such as temperature, pH value, templates, and magnetic or flow fields, is important to achieve ordered nanostructures [9-14]. Although these methods can increase the efficiency of the self-assembly, they can also complicate the fabrication process further, sometimes even more than the top-down approaches. Therefore, there is still a need for simple methods to synthesize monodisperse nanoparticles and to modify the surface properties to fully exploit the advantages offered by self-assembly.
Biomolecules, such as peptides and proteins, have been demonstrated to be useful in the synthesis and self-assembly of inorganic nanostructures [15,16]. Herein, we have investigated the utility of a silica-binding peptide (SiBP) in the single-step synthesis and self-assembly of SiO2 nanoparticles into ordered 3D structures. The SiBP is a member of the "solid-binding peptides" family. Solid-binding peptides are designed to have strong and often specific binding affinity to solid surfaces [17,18]. Because of their interactions with solid surfaces, these peptides have been shown to be able to functionalize nanostructures, catalyze the formation of nanostructures, and modify the nucleation, growth and self-assembly processes [19-24]. For this study, we have selected a second-generation SiBP, which was developed using a bioinformatics knowledge-based all-against-all comparison of first-generation SiBPs identified via phage display [25]. The SiBP used in the study was selected because of its high affinity to SiO2 and the presence of nucleophilic (serine) and basic (arginine) amino acids. The hypotheses of this study were as follows: (1) The basic serine and arginine residues in the SiBP can facilitate hydrolysis of the precursor molecules and, thus, catalyze the synthesis of SiO2 particles. (2) The affinity of the SiBP to SiO2 can narrow down the size distribution of the particles through a capping agent-like effect. (3) The SiBP can increase the efficiency of the self-assembly by modifying the net surface charge of the particle.
To test these hypotheses, we have synthesized SiO2 particles with the Stöber method using the SiBP as the only catalyst or in combination with NH3. The reaction kinetics were monitored via measuring the optical density (OD) with UV–vis spectroscopy and the conversion of substrate via gas chromatography coupled with mass spectroscopy (GC–MS). Size and net surface charge distribution of the particles were determined with dynamic light scattering (DLS). The efficiency of the self-assembly was evaluated with scanning electron microscopy (SEM), UV–vis spectroscopy, and qualitative visual demonstration.
Reaction kinetics were studied via OD measurements of the particles and GC analysis of tetraethyl orthosilicate (TEOS). OD profiles of the particle formation are shown in Figure 1a, and representative GC spectra of the control group are shown in Figure 1b. No particle formation or TEOS hydrolysis was observed in the negative control (no catalyst) group within the measurement timeframe. As seen from the OD profiles (Figure 1a) and TEOS conversion rates (Figure 1c), distinct profiles were observed when SiBP was added alone or in combination with NH3.
SiBP was able to hydrolyze TEOS and produce SiO2 particles when used alone. The increase in peptide concentration resulted in a dose-dependent increase in the reaction rate and yield (Figure 1d). Despite the same amount of precursors added to all reactions, the yields of the reactions with SiBP alone at plateau were lower compared to reactions containing NH3. Possible reasons of this observation will be discussed below.
No significant differences were observed regarding particle size or morphology depending on the SiBP concentration when SiBP was used alone as catalyst (Figure 2). At all concentrations, 17–20 nm particles were obtained with polydispersity indices (PDI) of 0.027, 0.050, and 0.010 for 0.04, 0.40, and 1 mM SiBP, respectively, indicating highly monodisperse (PDI < 0.080) particles. At all concentrations, the particles were organized into a branched fibrillar network, which is characteristic to the gel state of colloidal SiO2 (Figure 2) [26]. Colloidal gels are formed when colloidally suspended particles form a branched fibrillar network of particle strands through interparticle attractions. Under alkaline conditions, electrostatic repulsion between the SiO2 particles prevents formation of interconnecting particle strands. However, the presence of a cationic emulsifier allows for stable interparticle interactions and coagulation of the particles into interconnected particle strands [26]. Our findings indicate that, when used alone, the positively charged SiBP can also act as a cationic emulsifier resulting in the branched fibrillar networks observed by SEM.
In this aspect, when used alone, the SiBP mimicked the in vitro behavior of biosilicification-related proteins (BSRPs), such as silicateins and silaffins. BSRPs facilitate the formation of inorganic silica structures in marine organisms [27,28]. It has been reported that isolated silicateins and silaffins or certain repeating motifs of these proteins can facilitate silica precipitation in vitro [29-32]. The catalytic activity of these proteins is thought to be similar to the serine–histidine–aspartic acid (SHD) catalytic triad [33,34]. In this model, a hydrogen bond between serine and histidine increases the nucleophilicity of serine. Aspartic acid stabilizes the favorable orientation of histidine. Then a nucleophilic attack by serine on the Si–O bond of the precursor molecule results in a Ser–O–Si(OR)3 transitory complex. The hydrolysis is completed by the addition of water, separating the protein and the hydrolyzed precursor molecule, and the release of ethanol.
Although SiBP contains an N-terminal serine and two arginine residues, it does not contain histidine, aspartic acid, or another residue that can act as a H bond acceptor for serine. However, serine residues can form hydrogen bonds among themselves. Therefore, one can speculate that a hydrogen bond formed between the serine residues of two peptide molecules can increase the nucleophilicity. If this is the case, the nucleophilic attack of serine can facilitate hydrolysis of TEOS. However, a second and more likely speculation is that the SiBP mediates the hydrolysis through arginine residues. Arginine is a very strong proton acceptor with a side chain pKa of 13.80 [35]. Therefore, locally increased concentrations of OH− by arginine could facilitate the hydrolysis of silicon alkoxides, since OH− is also a potent nucleophile. Future studies where serine and arginine residues of the SiBP are substituted could help to elucidate the nature of the catalytic activity of SiBP.
As mentioned above, the yield of the reactions with SiBP alone were lower compared to reactions including NH3 despite the same amount of initial precursor molecules (Figure 1c). A probable reason of this observation is the high affinity of the peptide to SiO2 [25]. As colloidally stable silica particles start to form, the peptide starts to adhere to the surface of the particles, effectively being removed from the solution before all the precursor molecules available are hydrolyzed.
When NH3 was added, the SiBP had a negligible effect on the reaction rate and the yield (Figure 1c), indicating that SiBP contributed very little to the catalytic process. This can also be the result of high affinity of SiBP to SiO2. Since particle formation occurs relatively fast when NH3 is added, it is possible that SiBP binding to the particles results in the negligible effect on reaction rate and yield.
All groups containing NH3 yielded spherical submicrometer particles characteristic to the Stöber method (Figure 3a–d). However, a decrease in average particle size and size distribution was observed with increasing SiBP concentrations (Figure 3e,f). The PDI for the NH3 + 1 mM SiBP was lower compared to other groups containing NH3, indicating a narrower size distribution. However, the other groups also yielded monodisperse distributions with PDIs below 0.080 (Figure 3a–d). The second hypothesis of the study is that because of its high affinity to SiO2, SiBP can form a dense layer on the particle surface that improves the stability of nanoparticles [36]. These observations support this hypothesis, as smaller and more narrowly distributed particles were obtained with higher SiBP concentrations. Capping agents are not only used to regulate growth and size of colloidal nanoparticles, but also to control the physicochemical or biological characteristics. As will be demonstrated below, in addition to the size of the particles, the SiBP also changes the surface charge of the particles, resulting in improved self-assembly.
An interesting and distinct OD profile was observed in the NH3 + 1 mM SiBP reaction, which started with a fast increase reaching to a peak higher than the plateau of the other groups after approximately 40 min. This was followed by a steep decrease falling back to the plateau of the other groups after approximately 20 min (Figure 1a). This was at first thought to be the result of an error in experimental methods or measurement. The reactions were repeated with freshly prepared solutions and different brands of 96-well plates, but the distinct profile was observed in each repetition. To further investigate this interesting profile, samples were collected from the NH3 + 1 mM SiBP reaction and the NH3 only reaction in the middle of the steep increase (20 min), at the peak point (45 min), and in the middle of the steep decrease (55 min) (Figure 4a). SEM analysis showed that very different particle formation regimes occurred in the presence and absence of the SiBP. After 20 min, NH3 + 1 mM SiBP yielded a high amount of monodisperse particles of approx. 10 nm (Figure 4b), while NH3 alone yielded polydisperse particles of 10–40 nm (Figure 4c). After 45 min, NH3 + 1 mM SiBP yielded a mixture of irregularly shaped particles (Figure 4d; arrows), clusters of small 10 nm particles (Figure 4d; asterisk), and larger particles between 30 and 50 nm, while NH3 alone yielded spherical particles of 20 to 50 nm (Figure 4e). After 55 min, NH3 + 1 mM SiBP yielded particles of 60–100 nm. Small particle clusters or irregular particles were not observed (Figure 4f), while NH3 alone yielded spherical particles of 100 to 130 nm (Figure 4g).
Based on the OD and SEM observations, SiBP seems to drastically change the particle formation/growth regime above a threshold concentration. In the reactions of NH3 alone, NH3 + 0.04 mM SiBP, and NH3 + 0.4 mM SiBP, the growth regime followed the classical aggregative growth and monomer addition model [37]. According to this model, at the early stages of the reaction, the dominant regime is homogeneous nucleation of SiO2 particles. The growth continues by coalescence and Ostwald ripening of the particles. Growth by coalescence and Ostwald ripening is a fundamental process that plays a dominant role in nanoparticle formation. In coalescence, two or more particles combine to form a larger particle, whereas in Ostwald ripening, small particles dissolve in a solution and redeposit to form large masses. The process is mainly driven by the differences in chemical potential and surface energy between particles with different size and shape. In SiO2 synthesis, smaller particles with high surface energy dissolve via cleavage of siloxane bonds on the surface. The released silicic acid is then deposited onto particles with larger radius. Evidence of coalescence was observed in our study as well (shown by the arrows in Figure 3 and Figure 4). At later stages of the reaction, when the precursor concentration drops below the nucleation threshold, the dominant regime becomes growth by monomer addition to the surface of the particles.
At 1 mM concentration, however, the SiBP alters this profile and delays the aggregation of early approx. 10 nm particles into larger particles (Figure 4b). The most likely mechanism for this is the SiBP binding on the surface of the particles and, in turn, stabilizing the early particles. The resulting high number of small particles with very large surface area results in the high OD at 45 min, which is higher than the plateau of the positive control group (Figure 4a). When the fraction of the peptide to primary particles exceeds a critical value, the primary particles rapidly aggregate into larger particles, which results in the rapid drop in the OD (Figure 4a,d). Further studies are underway to take advantage of this interesting effect in synthesizing colloidally stable approx. 10 nm SiO2 particles at high volume fractions.
The effect of the SiBP on the self-assembly of the as-grown particles was investigated via SEM and UV–vis spectroscopy. Single-layer and multilayer assemblies were investigated by using different dilutions of the as-synthesized particles.
SEM imaging showed that the particles from the NH3 + 1 mM SiBP reaction assembled into ordered single-layer (Figure 5a) or multilayer (Figure 5c) opal-like structures. Opal is a naturally occurring mineraloid with silica as the principal chemical constituent. The optical behavior of iridescent opal is a result of the regularly stratified structure of silica particles, in which the alternate layers differ in refractive index. The periodic difference in the refractive index creates photonic band gaps, in which certain wavelengths of the light cannot propagate, depending on the size of the periodic structures and the differences in the refractive indices. These structures are referred to as photonic crystals and can be manufactured synthetically for various optical applications. Similar optical behavior of the opal-like structures formed in this study will be demonstrated and discussed below.
The fast Fourier transform of the SEM images revealed a hexagonal close-packed structure (insets in Figure 5a,c). In fact, the tendency of the particles formed with NH3 + 1 mM SiBP to assemble into ordered structures was visible on samples not prepared by vertical deposition but simply by dripping on a surface and vacuum drying (Figure 3d). The particles formed in the reaction with NH3 alone assembled into less ordered single-layer (Figure 5b) or multilayer (Figure 5d) structures.
In UV–vis spectra of the self-assembled particles, a broad peak was observed around 611 nm with NH3 only, while with NH3 + 1 mM SiBP, a narrower and stronger peak was observed around 457 nm (Figure 5e). The position and the width of the absorbance peaks in the UV–vis spectroscopy are determined by the Bragg diffraction of the light in photonic crystal structures and depend on the particle size, extent of the periodicity (i.e., quality of the assembly), and the angle of the incident light. The position of the absorbance peak can be calculated using Bragg's law (Equation 1):
where λ is the wavelength of the peak absorbance, d is the diameter of the particles, f is the packing factor (0.74 for hcp structures), n is the refractive index (1.46 for SiO2 [38] and 1 for air), and θ is the angle of the incident light. Using this formula and the average particle sizes obtained from the particle size analysis, the absorbance peak was calculated to be 593 nm for the NH3-only reaction and 453 nm for the NH3 + 1 mM SiBP reaction. The slight difference between the calculated and measured Bragg maxima is likely due to the hydrodynamic radius measured in particle size analysis being slightly larger than the actual size of the particles. Considering this, Figure 5e shows that the calculated and measured values agree well. The broader and weaker peak observed with the NH3-only group is the result of the less ordered assembly and broader particle size distribution. This is better observed when the measured λ values are fitted to the theoretical Bragg reflection maximum (see Supporting Information File 1, Figure S4).
The improvement in the self-assembly by the SiBP is likely achieved by reducing the negative surface charge of the SiO2 particles. The surface of the SiO2 particles is negatively charged above pH 2–3 [39,40]. Therefore, the as-synthesized particles exhibit little cohesion because of electrostatic repulsion. Usually, a post-assembly sintering process or long waiting periods [41] are required to improve the assembly. When the positively charged SiBP binds to the particle surface, it reduces the negative charge of the particle surface (Figure 5f). The particles synthesized with NH3 only had a ζ potential of −44.90 mV, while the particles synthesized with NH3 + 1 mM SiBP had a ζ potential of −30.87 mV. At the meniscus where the solvent dries out, the decreased surface charge reduces the distance between the particles, enabling them to assemble into more ordered and close-packed structures.
Angular dependence of the Bragg reflections and the uniformity of the assembled particles from the NH3 + 1 mM SiBP group was qualitatively investigated by assembling the particles on the inner surface of a cylindrical beaker by letting the reaction solution evaporate under vacuum. Following the curvature of the surface, different colors were observed depending on the angle of the incident light (Figure 6a). When the beaker was rotated under the same angle of incident light, the observed colors did not change, indicating a long-range uniform assembly. Side-by-side visual comparison of the uniformity of the self-assembly from the NH3 alone and NH3 + 1 mM SiBP reactions are demonstrated in Figure 6b.
We investigated the utility of a silica-binding peptide (SiBP) on SiO2 nanoparticle formation and self-assembly. We demonstrated that the SiBP can function in multiple ways in the synthesis and assembly processes (see Supporting Information File 1, Figure S5).
The first hypothesis of the study was accepted as when used alone, the SiBP was able to catalyze the hydrolysis of precursor molecules and precipitate approx. 20 nm SiO2 particles organized in a branched fiber network. We propose that the catalytic activity is mediated either by the serine residue or the arginine residues, or a combination of both.
The second hypothesis of the study was partially accepted as when used alone, the SiBP had no effect on particle size regardless of the peptide concentration. However, when used in combination with NH3, the SiBP resulted in smaller and more uniform particles with increased peptide concentration.
The third hypothesis of the study was accepted as the SiBP improved the self-assembly behavior by adhering on the particle surface and reducing the negative surface charge.
The results presented here provide a single-step biomimetic route to synthesis and assembly of SiO2 nanoparticles into gels or opal-like ordered structures.
The SiBP was commercially synthesized (Genscript, NJ, USA) at <98% purity via 9-fluorenylmethoxycarbonyl (F-moc) chemistry methods. The amino acid sequence and physicochemical properties of the SiBP are shown in Table 1.
Table 1: Amino acid sequence and physicochemical properties of the SiBP.
|Amino acid sequence
|Molecular weight (Da)
|Isoelectric point
|Grand average of hydropathy
|Net charge
The particles were synthesized based on the method described by Stöber and co-workers [42]. Tetraethyl orthosilicate (TEOS) (Alfa Aesar, Ward Hill, MA, USA) was used as the precursor, a 1:1 mixture of ethanol/deionized water was used as the solvent, and NH3 (Thermo Fischer Scientific Inc, Waltham, MA, USA) was used as the catalyst. Briefly, the Stöber process involves hydrolysis of an alkoxysilane precursor, such as TEOS, in alcohol (typically methanol or ethanol) in the presence of a catalyst. This is followed by ethanol or water condensation polymerization of the hydrolyzed precursor. Once colloidally stable nuclei form, growth ensues through monomer addition or coalescence and Ostwald ripening. The reactions involved in the Stöber process are described as follows:
where Equation 2 is the ionization of the ammonia, Equation 3 is the hydrolysis of the alkoxysilane, Equation 4 is polymerization via alcohol condensation, and Equation 5 is polymerization via water condensation.
The reaction solvent was prepared by mixing equal volumes of gradient grade (≥99.9%) ethanol (Sigma-Aldrich, MO, USA) and deionized water (or a solution of the peptide prepared in deionized water). Then TEOS (≥99.0%) (Sigma-Aldrich, MO, USA) was added to a final concentration of 0.45 M and mixed rigorously for 10 min. The reaction solutions were transferred to a 96-well plate in triplicates for each group. The reactions were started by adding NH3. The reactant contents for different groups are given in Table 2. In groups 1, 2, and 3, no NH3 was added to investigate whether SiBP had a catalytic activity by itself to precipitate SiO2 particles. In groups 4, 5, and 6, 0.2 mM NH3 (prepared from 25% ammonium hydroxide (Merck & Co., NJ, USA)) was added in combination with different concentrations of the peptide to examine whether the peptide had a synergistic effect on the formation of SiO2 particles.
Table 2: Reaction contents for the different conditions studied.
|0.2 mM NH3
|(−) control
|(+) control
|0.2 mM NH3
|group 1
|0.04 mM SiBP
|group 2
|0.40 mM SiBP
|group 3
|1.00 mM SiBP
|group 4
|0.04 mM SiBP + 0.2 mM NH3
|group 5
|0.40 mM SiBP + 0.2 mM NH3
|group 6
|1.00 mM SiBP + 0.2 mM NH3
The reaction rates were monitored by measuring the optical density (OD) of the particles at 445 nm (Varioskan Flash micro-plate reader, Thermo Fischer Scientific Inc, Waltham, MA, USA) and by monitoring the hydrolysis of TEOS via gas chromatography coupled with mass spectrometry (GC–MS) (TSQ Duo Triple Quadrupole, Thermo Fischer Scientific Inc., Waltham, MA, USA). OD measurements were done at room temperature and constant mixing at 300 rpm. GC–MS analyses were done with a fused silica capillary column a (Restek 14623, Thermo Fischer Scientific Inc, Waltham, MA, USA), 150 °C injection temperature, 250 °C detection temperature, 1 µL injection volume, and 1 mL/min He flow rate. Dionex Chromeleon 7.2 software (Thermo Fischer Scientific Inc., Waltham, MA, USA) was used for the quantitative analysis of GC–MS data. The reaction rate of TEOS hydrolysis was calculated according to first-order reaction kinetics [43].
Measurements were performed using a Zetasizer Nano ZS90 (Malvern Panalytical Ltd, Malvern, UK). As-synthesized particles were diluted to 1:10 in the reaction solvent (1:1 ethanol/deionized water) and dispersed by sonication for 10 min. Readings were taken at room temperature. Malvern DTS software v.5.10 (Malvern Panalytical Ltd, Malvern, UK) was used for data processing and analysis. Particle size distributions are reported based on intensity weighted averages and expressed as mean diameter ± SD. Each reading was repeated three times and readings were done with three runs (3 min/run).
Colloidal particles were synthesized in the same way as described in Table 2 with a final volume of 10 mL. The particles were assembled by a vertical deposition method. For SEM analysis, the particles were assembled on regular microscope cover slides. For UV–vis spectroscopy analysis, the particles were assembled on quartz slides. One end of the slide was attached to a NE-1002X micro-fluidics automatic syringe pump (New Era Pump Systems Inc., Farmingdale, NY, USA) and the other end was vertically dipped into the as-synthesized colloidal solution of particles (see Supporting Information File 1, Figure S3) The syringe pump was run at a speed of 1.50 µm/s until the covers were moved completely out of the colloidal solution.
To analyze as-synthesized particles, 50 µL aliquots of the reaction solutions were placed on standard microscope cover slides. The excess liquid was removed by absorbing on a clean absorbent paper. To analyze self-assembled particles, cover slides described in the previous section were used. The slides were dried under vacuum and then adhered onto an aluminum sample holder using carbon tape. The samples were coated with platinum for 30 s using an EM ACE200 vacuum coater (Leica Microsystems GmbH, Wetzlar, Germany). All SEM analyses were performed using a SU5000 SEM (Hitachi, Japan) at 10 kV accelerating voltage.
The particles were assembled on quartz slides by vertical deposition as described before. UV–vis absorbance analysis was made using a T80+ UV–vis spectrophotometer (PG Instruments Ltd., Leicestershire, UK).
Supporting Information features mass spectra of the reaction with NH3 only at different time points, mean OD values collected from three separate syntheses, a schematic representation of the vertical deposition method used in the study, the fit of measured λ values to the theoretical Bragg reflection maxima, and a schematic representation of the multiple roles of the SiBP.
|Supporting Information File 1: Additional experimental data.
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