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Category Archives: Nanotechnology

How Can Nanotechnology be Used in Wheat Biofortification? – AZoM

The growing world population needs a secure supply of food. Biofortifying staple foods is one way to solve challenges in the food industry. A paper recently published in Biology has considered the use of nanotechnology to fortify wheat, a key global crop.

Study:Insight into the Prospects for Nanotechnology in Wheat BiofortificationImage Credit:maxbelchenko/Shutterstock.com

Biofortification is the process of deliberately increasing the nutritional value of a food by agronomic methods, breeding, and biotechnology. It differs from conventional fortification in that it aims to increase the nutritional value of a plant during its growth rather than manually adding vitamins, minerals, and trace elements during processing.

Biofortification has important implications for reaching populations that have no or limited access to traditional supplements and fortification methods. It provides public health benefits with minimal risk to the health of populations and individuals.

Different strategies used for wheat biofortification. The left side of the figure explains the wheat biofortification strategies other than nanobiofortification and their drawbacks. The right side of the figure provides a summary of nanobiofortification including its target, entry points of nanofertilizers in plants, and the advantages of nanomaterials (Feiron, Znzinc, CuCopper, NPKnitrogen, phosphorus and potassium).

Wheat is consumed by a sizable proportion of the worlds population. Increasing its nutritional value will tackle issues with malnutrition in growing populations. In recent decades, several biofortification strategies have been attempted with varying degrees of success. Conventional breeding, agronomic biofortification, and transgenic approaches have all been employed to provide nutritional benefits.

Whilst these approaches have had their successes, there have also been major drawbacks to them. There are regulatory barriers to the field, even though the biofortification methods themselves are well-developed. Genetic biofortification is limited by restrictions in the available targeted biological gene pool as well as being time-consuming. Agronomic biofortification suffers from problems with available fertilizers. Conventional plant breeding takes time.

Nanobiofortification has gained traction in recent years as a viable alternative to contemporary genetic and agronomic methods. Using nanotechnology can circumvent time issues, resource requirements, and environmental risks.

One main benefit is the targeted delivery of fertilizer in the required amount, increasing nutrient uptake and preventing the leaching of potentially harmful inorganic fertilizer into fragile ecosystems. Nanomaterials used for this purpose vary in efficiency due to size, composition, chemical features, and the plant they are targeted for.

An overview of the different methods used for Wheat Nanobiofortification, types of applied nanomaterials, and their outcome in the form of fortified nutrients. (Abbreviations in the figure: NPs- nanoparticles; ZnOzinc oxide; dexdextran; Fe2O3ferric oxide; Fe3O4ferrosoferric oxide; NPKnitrogen, phosphorus, potassium; CNPchitosan nanoparticles; Bboron, Feiron, Znzinc; Sisilicon; Pphosphorus; Cucopper).

Nanotechnology is utilized in biofortification in numerous ways. Nanobiofortification systems can introduce nutrients into soil in a controlled release rate. Their ability to provide wide coverage and efficient absorption support plant development and environmental safety, compared to agronomic fertilizers.

Currently, three different types of nanomaterials are being employed for biofortification: nanoscale coating fertilizers, nanoscale additives fertilizers, and nanoscale fertilizers (NFs.) Nanomaterials used in these fertilizers can be classified as polymeric nanomaterials, ceramic nanomaterials, and metallic nanomaterials.

The nanomaterials used in nanofertilizers can be grouped into macronutrient fertilizers, micronutrient fertilizers, plant growth-stimulating NFs, and nanomaterial-enhanced fertilizers. This grouping is due to the nutritional benefits the nanoscale systems confer upon the target plant. The research provided many perspectives on nanobiofortification methods, some of which will now be discussed.

Several micronutrients are essential for wheat, including zinc, the deficiency of which causes necrosis, stunted growth, decreased seed quality, and decreased yield. Traditionally micronutrient enhanced composite NPK fertilizers do not always provide wheat with the necessary concentration of micronutrients.

The small size and large surface area of nanoparticles significantly increase the bioavailability of the micronutrients. Zn and Fe enhanced NFs are of particular focus due to the deficiency of these micronutrients in the world population.

Important macronutrients including calcium, magnesium, and nitrogen are encapsulated or inserted into macronutrient NFs. The addition of these macronutrients to NPK fertilizers in the amounts necessary increases the amount of fertilizer needed, which presents a serious ecological risk. Nanotechnology-based delivery systems are more efficient at delivering macronutrients as well as being more sustainable.

There are different routes by which nutritionally enriched nanomaterials can be introduced into wheat plants. Seed priming, soil fertilization, and foliar fertilization can all be employed. Studies have been conducted into all three routes, demonstrating the benefits of NPs.

In seed priming, seeds are treated before planting. Seed priming with nanoparticles improves metabolism and delivers nutrients more effectively than conventional methods. Both soil and foliar fertilization show increased nutrient uptake and yield using NPs.

Biofortification with nanoparticles shows huge promise for the future. Optimized delivery can save costs and allow so-called Precision farming. Whilst NFs have well-discussed health benefits, controlling their supply and accumulation needs to be carefully monitored, as their long-term health and environmental risks are not properly understood.

Although there are some reports of toxicity in nanomaterials used for agricultural purposes, they remain one of the best solutions to the challenges facing modern agriculture and feeding a growing world population.

Kahn, M.K et al. (2021) Insight into the Prospects for Nanotechnology in Wheat Biofortification [online] Biology 10(11) 1123 | mdpi.com. Available at:https://www.mdpi.com/2079-7737/10/11/1123/htm

Disclaimer: The views expressed here are those of the author expressed in their private capacity and do not necessarily represent the views of AZoM.com Limited T/A AZoNetwork the owner and operator of this website. This disclaimer forms part of the Terms and conditions of use of this website.

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How Can Nanotechnology be Used in Wheat Biofortification? - AZoM

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Spanish nanotechnology could be used on new Euro banknotes – Euro Weekly News

According to the Spanish National Research Council (CSIC), the next batch of Euro banknotes could be made with new materials and nanotechnology developed in Spain. Presented at the CSIC headquarters in Madrid, they claim that the objective of the new notes is to improve their safety and durability, while increasing the quality and sustainability.

A research team from the Madrid Institute of Materials Science (ICMM-CSIC), is responsible for this development, and it works in collaboration with the Bank of Spain. Funding for this comes from the Eurosystem the monetary authority of the Eurozone.

The Bank of Spain has been advising and collaborating in the CSIC research so that the results of the project can be applied to euro banknotes. This project, which began in October 2019, is due to conclude its first phase in 2022, and is subject to strict confidentiality requirements.

Tasked with safeguarding the integrity and security of euro banknotes is The European Central Bank. With this objective, the European Central Bank, and all national central banks, such as the Bank of Spain, try to incorporate increasingly advanced technologies to achieve better banknotes.

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Spanish nanotechnology could be used on new Euro banknotes - Euro Weekly News

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New publication in Nature Nanotechnology on safe engineering of therapeutic cells with photothermal nanofibers – ELIS

Our group has demonstrated the possibility to use photothermal nanofibers and laser irradiation as a new methodology to deliver gene editing effector molecules in living cells. We have demonstrated broad applicability in different therapeutic cells, such as T cells and stem cells, with a variety of effector molecules, such as siRNA and CRISPR/Cas9 technology. Importantly, we were able to show that the quality of the therapeutic cells is significantly better as compared to commercially available technologies, leading to better therapeutic potency of the final cell product.

More information can be found in the original publication (https://www.nature.com/articles/s41565-021-00976-3) or in this blog article: https://bioengineeringcommunity.nature.com/posts/safe-engineering-of-therapeutic-cells-with-photothermal-nanofibers

Xiong R., Hua D., Van Hoeck J., Berdecka D., Lger L., De Munter S., Fraire J., Raes L., Harizaj A., Sauvage F., Goetgeluk G., Pille M., Aalders J., Belza J., Van Acker T., Fernandez E.B., Si T., Vanhaecke F., De Vos W., Vandekerckhove B., van Hengel J., Raemdonck K., Huang C., De Smedt S.C., Braeckmans K. Photothermal nanofibers enable safe engineering of therapeutic cells. Nat. Nanotechnol. DOI: 10.1038/s41565-021-00976-3 (2021).

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Seven spooky things that people say are in the COVID-19 vaccines but definitely arent – Austin American-Statesman

Samantha Putterman| PolitiFact.com

Just in time for Halloween, claims about whats lurking in the COVID-19 vaccines are getting wilder and spookier.

Theres the "transhumanism nanotechnology" ingredient that will apparently turn all of us into copies of the Terminator. Theres also "graphene oxide," a material that some claimed will straight up kill you. Why didnt Michael Myers think of that?

Rest assured, weve fact-checked this, and its all fake gore.

Heres a look at seven scary things that are not in the vaccines and, as a treat, a list of the rather helpful things that are.

Aluminum. They use it in pickup trucks, food containers and antiperspirants. But in the COVID-19 vaccines?No.

The U.S. Centers for Disease Control and Prevention reports that small amounts of aluminum specifically aluminum salts have been used in vaccines since the 1930s as an adjuvant, which helps elicit a stronger immune response from the body.

But none of the three COVID-19 vaccines currently being used in the U.S. Pfizer, Moderna and Johnson & Johnson, all of which have publicly accessible ingredient lists contain any aluminum.

As for other vaccines,research has showntheir levels of aluminum are so low that they can't easily be absorbed by the body, let alone the brain. There has been no evidence of the aluminum in vaccines causing illness or developmental disorders.

While your cell phone can help Big Brother, or your mother, locate you via 5G networks,the vaccines cannot.

Yet some social media users likened the bubbles of fat in the vaccine to the sort of tracking microchips implanted under a pets skin. Thats a lot of creepy nonsense.

There is no evidence that the COVID-19 vaccines contain technology similar to pet microchips. The lipid nanoparticles used in some of the vaccines are called "nanoparticles" because they are very, very small. They have nothing to do with 5G networks or tracking technology.

This baseless conspiracy theory says that the shots include a technology that changes "what it is to be human."The vaccines dont contain any such thing.

"None of the vaccines contain nanotechnology of any sort, let alone 'transhumanism nanotechnology, which isnt even a thing," Mark Lynas, a visiting fellow at the Alliance for Science and Cornell University, told PolitiFact.

"Nano," as we said earlier, is a term widely used to describe things that are very tiny, and scientists use the prefix more specifically to refer to things on the scale of individual atoms.

A chilling, grainy black-and-white image being shared on social media has been described as a "Trypanosoma Parasite" purportedly observed in Pfizers COVID-19 vaccine. Several variants of the parasite, internet users claimed, are lethal and are one of many causes of acquired immune deficiency syndrome or AIDS.

This is erroneous, on all counts.

Dr. Bobbi Pritt, the director of clinical parasitology at the Mayo Clinic, told us the blurry image likely represents an out-of-focus non-cellular component of the vaccine and doesnt show a Trypanosoma cruzi or any other parasite.

As for the claim that this particular parasite causes AIDS, thats also wrong, she said.

"The only thing that causes AIDS is an infection with the human immunodeficiency virus," she said, "and this cannot be acquired through the Pfizer vaccine."

A popular video claimed that the Pfizer vaccine contains "particles that could germinate and cause illness" and that vitamin supplements could stop this from happening. You can bet someone is selling those supplements online.

Theres no truth to this one either. The ingredients for Pfizer vaccines are chemical components not living organisms.

"Contamination with spores or other microbial material can theoretically happen during production of any biologic, including vaccines," said Volker Mai, associate professor in the epidemiology department at the University of Florida. "However, quality control is extensive and monitoring occurs continuously. Thus, it is highly unlikely that any contaminated batch would make it into the market."

A vaccine turning you into the Marvel villain Magneto? That sounds terrifying (unless thats what you wanted.) But dont worry, the COVID-19 vaccineswont make you more attractive to magnets.

Social media users have shared videos that appear to show magnets sticking to peoples arms where they say they were injected, and claimed this as proof the shots have microchips in them.

But medical experts called the claim utter nonsense.

Al Edwards, an associate professor in biomedical technology at the University of Reading in England,told Newsweekthat because vaccines ingredients are some of the same things that are in the human body, "there is simply no way that injecting a tiny fragment of this material" could make it respond to a magnet. "Most food is made of similar molecules, and eating food doesnt make people magnetic," he said.

A similar claim cites a video showing what looks like small balls connecting and growing on their own. The disturbing clip was described as the COVID-19 vaccines reaction once it hits the bloodstream.

Thats wrong.The video was actually from a 2015 science experiment by the Stanford Complexity Group, an initiative to bringcomplexity scienceto a wider audience, that shows self-organizing wires, which is still weird.

One more time for the people in the back: None of the COVID-19 vaccines contain microchips or metals.

An incendiary video that speaks of murder claims that Pfizers COVID-19 vaccine is dangerously packed with something called graphene oxide.

A Pfizer spokesperson told PolitiFact that while graphene oxide a material made by the oxidation of graphite is used in some vaccines, it is not used at Pfizer andis not in its COVID-19 vaccine.None of the listed ingredients is another name for graphene oxide, and the material doesnt appear in ingredient lists for the Moderna and Johnson & Johnson vaccines.

The real ingredients are much less likely to keep you up at night.

All three manufacturers of the approved vaccines in the U.S.,Pfizer,ModernaandJohnson & Johnson, have shared their ingredients.

For Moderna and Pfizer, the active ingredient is messenger RNA, which carries genetic information about the coronavirus to the body's cells to help them recognize and produce antibodies against it. Theres no evidence that mRNA is dangerous, and scientists say the material is broken down by the body within days.

Pfizers inactive ingredients include lipids, salts, sugar and saline solution.

The lipids encase the mRNA, the salts help keep the pH, or acidity, of the vaccine close to that of a persons body; and the sugar safeguards the lipids when theyre frozen and stops them from sticking together,according to MITs Technology Review, which spoke with experts to help decode the contents.

Before injection, the vaccine is mixed with the saline solution, just as many intravenously delivered medicines are, the report said.

Modernas ingredient list is very similar, and includes the lipids, salts and sugar, with a slight difference, including the solution for the injection, that may explain the different storage needs for each.

The Johnson & Johnson vaccine uses a disabled adenovirus, rather than mRNA, to deliver instructions to produce the coronavirus spike proteins and activate the immune system. The other ingredients are alcohol, citric acid, salts and sugars.

Afull list of ingredientsfor all the vaccines is available on the CDC website.

PolitiFact,The COVID-19 vaccines do not contain aluminum, March, 22, 2021

PolitiFact,No, COVID-19 vaccines do not contain nanoparticles that will allow you to be tracked via 5G networks, March 12, 2021

PolitiFact,Transhumanism nanotechnology COVID-19 vaccine conspiracy theory is Pants on Fire, Oct. 21, 2021

PolitiFact,No, this isnt a picture of a parasite in the Pfizer COVID-19 vaccine, Oct. 13, 2021

PolitiFact,No living organisms in the Pfizer vaccine, Oct. 15, 2021

PolitiFact,No, this isnt a video of a COVID-19 vaccine, Oct. 21, 2021

PolitiFact,No evidence of graphene oxide thats toxic in Pfizer COVID-19 vaccine, July 8, 2021

PolitiFact,No, these magnet videos dont prove the COVID-19 vaccines contain microchips, May 17, 2021

U.S. Food & Drug Administration,VACCINE INFORMATION FACT SHEET FOR PFIZER-BIONTECH COVID-19 VACCINE, Updated Oct. 20, 2021

U.S. Food & Drug Administration,FACT SHEET FOR MODERNA COVID-19 VACCINE, Updated Oct. 20, 2021

U.S. Food & Drug Administration,FACT SHEET FOR THE JANSSEN COVID-19 VACCINE, Oct. 20, 2021

MIT Technology Review,What are the ingredients of Pfizers covid-19 vaccine?, Dec. 9, 2020

U.S. Centers for Disease Control and Prevention,Appendix C: Ingredients included in COVID-19 vaccines, Updated Oct. 27, 2021

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Seven spooky things that people say are in the COVID-19 vaccines but definitely arent - Austin American-Statesman

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Advancing the Synthesis of Freestanding Nanostructures with 3D Printing – AZoNano

An international team of scientists based at Graz University, Austria, and the Center for Nanophase Materials Sciences in Oak Ridge National Laboratory, and the University of Tennessee in the United States recently published the most wide-ranging measurements of nanowires evolution and features seen to date. The team hopes their paper will form the basis for future research into synthesizing freestanding nanostructures with 3D printing.

Image Credit:TriMech/Shutterstock.com

A nanostructure is a structure that is too small to be seen with a conventional microscope but bigger than the molecular scale. Nanostructures have at least one dimension on the nanoscale, between 0.1 and 100 nm. (One nanometer nm is one-billionth of a meter, or 110 m).

Nanotextured surfaces like thin films have only one dimension on the nanoscale, thickness. The nanowires used for nanoscale 3D printing have two dimensions, but their length is much greater. Spherical nanoparticles have three dimensions on the nanoscale.

In each case, the nanostructures structure, surface, and materials behave according to the peculiar laws of quantum mechanics along whichever dimension or dimensions are measured on the nanoscale.

This means that we know very little about how nanostructures interact with the world, mechanically, chemically, or electromagnetically, compared to objects and structures on bigger scales of size.

We know even less about nanowires, according to the papers authors.

Nanowires were an early nanostructure of interest to nanotechnology researchers, who dubbed them quantum wires due to their strange quantum mechanical behaviors. They are structures whose diameter is on the nanoscale and whose length is at least 1,000 times greater than their diameter or width.

Carbon nanotubes somewhat overtook nanowires in terms of exciting nanostructures in two dimensions in recent years. They are made by rolling thin films into a hollow tube and have excellent mechanical strength and semiconductor properties.

These Nanostructures Are Hacking NaturePlay

Video Credit: Seeker/YouTube.com

The precise measurement of nanowires in the recent study, published in Additive Manufacturing, is important for making calculations and simulations of nanostructures. It is especially relevant to feed into machine learning and human design seeking to optimize nanowires applications.

Nanowires are synthesized with an additive manufacturing process, Focused Electron Beam Induced Deposition (FEBID). FEBID is a way to 3D print nanowires, enabling the direct-write fabrication of complex 3D objects onto practically any substrate material.

Understanding the precise shape, reactions, evolution, and other properties of nanowires is essential to understanding the physical properties of freestanding nanostructures 3D printed with FEBID technology.

Additive manufacturing builds 3D objects up, usually layer by layer, from a computer models instructions.

Additive manufacturing processes use material more efficiently, provide greater design freedom, and achieve higher degrees of precision than traditional subtractive manufacturing methods, which involve cutting a shape out of a block of material.

It is also is much faster than traditional manufacturing, avoiding the need for molds and guides and enabling manufacturers to switch from one product or component to another without reconfiguring anything.

Applying the concepts of 3D printing to nanotechnology brings similar advantages. 3D printing nanostructures can be faster, less wasteful, and more precise than other top-down fabrication methods.

3D printing is the only technology capable of synthesizing many nanostructures, primarily freestanding nanostructures. Brittle ceramics can be shaped into high-order 3D structures only with additive manufacturing, which may predict the future of compact sensor design, energy scavengers, and diagnostic devices.

Another miniaturized 3D printing technology that will be key in the future of manufacturing at the nanoscale is focused electron beam induced deposition (FEBID). FEBID creates freestanding nanostructures out of nanowires, and the precise measurements of nanowires in the recent research will contribute to FEBIDs ongoing commercial adoption.

FEBID is a direct-writing technique that can be used to 3D print freestanding nanostructures. The technique involves adsorbing a precursor material on a substrate surface and then dissociating it in focus on an electron beam.

Also known as electron beam induced deposition (EBID), FEBID uses the electron beam to decompose gaseous molecules and depose non-volatile fragments onto the substrate. The beam comes from a scanning electron microscope, leading to the exceedingly high degrees of spatial accuracy that make the technique suitable for nanotechnology.

Researchers developing the technique have been able to create the worlds smallest magnet, fractal nanotrees, nanoloops, and superconducting nanowires using FEBID so far. Having been under close scrutiny for the last decade or so, FEBID is now at a state of maturity that means it can start to see applications in research or commercial fields.

With the precise measurements of nanowires in the Additive Manufacturing paper, FEBID can now continue on its path towards wider adoption.

Remaining challenges remain, including improving methods for controlling metal content in FEBID structures and increasing the deposition yield. Future work in this field is likely to produce unexpected results for fundamental or application-driven researchers alike.

Continue reading: Carbon Nanotube Nanocomposite Ink for Additive Manufacturing.

Winkler, R. et al. (2021) Shape evolution and growth mechanisms of 3D-printed nanowires.Additive Manufacturing. Available at: https://doi.org/10.1016/j.addma.2021.102076

Berger, M. (2014) Nanotechnology and 3D-printing.Nanowerk. Available at: https://www.nanowerk.com/spotlight/spotid=37541.php

Huth, M. (2011) Focused Electron Beam Induced Deposition Principles and Applications.Beilstein-Institut. Available at: https://www.beilstein-institut.de/download/267/pdf_huth.pdf

Huth, M. et al. (2012)Focused electron beam induced deposition: A perspective.Beilstein J Nanotechnology. Available at: https://doi.org/10.3762/bjnano.3.70

Disclaimer: The views expressed here are those of the author expressed in their private capacity and do not necessarily represent the views of AZoM.com Limited T/A AZoNetwork the owner and operator of this website. This disclaimer forms part of the Terms and conditions of use of this website.

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Exotic magnetic states on the nanoscale – Nanowerk

Nov 05, 2021(Nanowerk News) An international research team, led by scientists from the EMPA (Zurich) and the International Iberian Nanotechnology Laboratory, which has researchers from the UPV/EHU, has succeeded in building chains of quantum magnets made of nanographene that capture the essence of one of the core models of quantum magnetism. The team's results (Nature, "Observation of fractional edge excitations in nanographene spin chains") have implications for understanding quantum magnetism on the nanoscale and may open the door to building quantum computers.Artistic rendering of a triangulene quantum spin chain adsorbed on a gold surface and probed with the sharp tip of a scanning tunneling microscope. While each triangulene unit (triangular nanographenes) has a total spin of 1, quantum correlations in the chain lead to spin fractionalization, such that the terminal triangulene units exhibit a spin of ½. (Image: EMPA)We are all used to the idea that simple units combine to form more complex structures. So atoms combine to form molecules, which in turn combine to form cells, and cells form tissue, ultimately giving rise to living beings.In the quantum world, this process may take place in reverse in such a way that the interaction between complex particles results in simpler particles. So, under certain circumstances, the interaction between electrons, indivisible particles with electric charge e, gives rise to the emergence of particles with charge e/3. This phenomenon is known as fractionalization.Quantum magic: fractionalizing spinsAll elementary particles have intrinsic properties such as mass or charge that are intuitive to us, and others such as spin, which can be visualised like a compass. However, unlike normal compasses, which can point in any direction, the spin of quantum systems is quantized, and can only assume a discrete set of values. For example, we say that the spin of an electron is ½ and can only take two values. Particles with spin 1 can take three values.In the 1980s the British physicist Duncan Haldane built a mathematical model for spin 1 particles in which the fractionalization of the spins took place. So when a one-dimensional chain of indivisible spin 1 particles interacted with their neighbours, they gave rise to the emergence of spin ½ particles on the edges of the chain.Like the magic trick in which the magician saws a person in half and pulls the two halves apart, the Haldane model allows spins 1 to be fractionalized and separated. It is one of the core models of quantum magnetism, and his work earned him the Nobel Prize in 2016.One-dimensional chains of magnetic molecules assembled from grapheneExperimental confirmation of this prediction was challenging for various reasons, chief among them being the fact that one-dimensional materials do not exist. Indirect evidence of the phenomenon of spin fractionalization in organometallic materials containing chains of magnetic atoms existed, but direct observation remained elusive.Now, however, that observation has been made by an international team of researchers, including the Ikerbasque researcher David Jacob of the Department of Polymers and Advanced Materials: Physics, Chemistry and Technology at the UPV/EHU; he has collaborated in this work with the INL, the University of Alicante, the EMPA in Zurich and the University of Dresden.To accomplish this difficult feat, the researchers combined organic chemistry techniques with ultra-high vacuum surface science in order to synthesise graphene molecules with spin 1 that form unidimensional chains. Using a tunnelling microscope, the team of researchers were able to study, with atomic resolution, the quantum states of the chain adsorbed on a gold surface, compare them with those predicted by the theory, and establish that the system did in fact behave like the Haldane model.In particular, in chains with a sufficiently high number of magnetic molecules, the researchers found Kondo resonances at the tips of the chain, a phenomenon that occurs when spin ½ particles interact with the electrons in a conductor such as gold.From one-dimensional chains to two-dimensional networks and quantum computersThe researchers say that this work "shows the potential for using nanographenes to form two-dimensional networks of nanomagnets, enabling predictions analogous to Haldane's to be confirmed, such as, for example, the existence of quantum states that would allow quantum computation to be carried out".

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