Many thanks to SWLing Post contributor, Adrian Korol, who shares the following press release from RAE and LRA 36 [note that this article has been machine-translated into English]:

New Short Wave Transmitter for LRA 36 Radio Nacional Arcangel San Gabriel

The Minister of Defense, Jorge Taiana, together with the Joint Antarctic Commander, Brigadier General Edgar Calandín, the President of Radio and Television Argentina SE (RTA), Licencida Rosario Lufrano, and the Executive Director of Radio Nacional, Alejandro Pont Lezica, participated in the act where this Monday (4-09) a short wave transmitter was donated for the radio LRA 36 “Arcángel San Gabriel”, located at the Esperanza Antarctic Base.

This incorporation -which occurs within the framework of a new collaboration between the Joint Antarctic Command (COCOANTAR) and the state company RTA- will allow the station to significantly improve the coverage and quality of the Antarctic station’s signal, maintain its international reach and fulfill the objective of disseminating the work of the Armed Forces and scientific personnel who work in the Antarctic bases of our country.

“This is an act of generosity, of solidarity with all Argentines, it is also an act of patriotism, because it will make it possible for all of us to listen to radio broadcasts from Antarctica,” said Taiana.

For the minister, “it is important for all Argentines to have in their heads and in their hearts that there is a part of our territory that is there, and that there are Argentines doing their work with great effort for a year on that continent.” , he claimed.

For his part, Lufrano said: “We have always said that our National Radio and Public TV should be broadcast by land, by sea, by air, and we are complying. It was an honor to be able to take these teams to the ARA “Almirante Irízar” icebreaker so that they could listen to all our programming, listen to our radio and watch our public television”.

In his turn, Brigadier General Calandín expressed: “For us it is a joy, an enormous satisfaction, to be able to show in this simple and emotional ceremony everything that Radio Nacional and the Ministry of Defense have done to contribute to the sovereign exercise of the antartida”.
And he pointed out: “This transmitter will allow us to arrive in a reliable way with everything that Argentine culture means in the Argentine sovereign exercise in Antarctica, it is a transcendental fact for us.”

During the act, the presence of Javier Gratz stood out, who is the donor of the transmitter equipment in a personal capacity, who informed that it is manufactured entirely in Argentina.

The donation was preceded by technical survey work carried out at the Esperanza Joint Antarctic Base by Alejandro Petrecca (Head of the National Radio Transmitter Plant in Pacheco) and Engineer Alejandro Alvarez, and driven by the relationship they maintain with RAE (Radio Difusión Argentina Abroad) entina.

The act was transmitted by AM 870 in duplex with the network of federal stations. The director of Radio Nacional, Alejandro Pont Lezica; the director of Rae, Adrián Korol; Marcelo Ayala (who traveled to Antarctica to make the federal panorama at the beginning of this year) and the conductors Mario Giorgi and Federica Pais.

LRA36 began its transmissions on October 20, 1979 and since then it has been operating uninterruptedly from the Esperanza Base in Argentine Antarctica. It is one of the most sought after stations by shortwave listening fans from all over the world, and its signal has been received in places as far away as Alasa, Iceland, Japan, most European countries, and Latin America.

It is the only public station that transmits from Antarctica in Short Wave, a characteristic that, added to the fact that it is operated by personnel from the Joint Antarctic Command, makes it unique in relation to the other National Radio stations, such as RAE.

During 2022, for the first time, the entire LRA36 team was made up entirely of women, a characteristic that is also repeated this year and, as anticipated on the spot, will be maintained in 2024.

Various areas of Radio Nacional, teachers from the National University of Quilmes, RAE and the Antarctic Joint Command participate in the training of those who make up the radio and the general coordination under the coordination of Mr. Juan Carlos Benavente.

Being a station with international scope, the objective of its programming is to disseminate Argentine culture, but above all the work of our Armed Forces in the Antarctic bases, scientific research work and, fundamentally, the support of the historical claim of sovereignty Argentina on the Antarctic territory.

The transmitter will have a power of 10 kW and it is expected to be working effectively during the 2024 Antarctic Campaign.

A bill that has sailed through the legislative process and is headed to the state Senate floor is a clever attempt to invent an alternative narrative for the perpetually floundering Los Angeles County Probation Department. Assembly Bill 695 would create a grant program that once funded would provide state money to L.A. County — and only L.A. County — for juvenile justice infrastructure.

The creative story line being used is that crumbling juvenile halls created dangerous conditions for youths and staff. Facilities were supposedly so outdated that state regulators had to step in and declare them unsuitable to house youths. The county Board of Supervisors wouldn’t act, so two county labor unions — the probation officers and the probation managers — took the initiative and drafted a bill to make state money available for replacement facilities and desperately needed refurbishment. Should the bill pass, grants will be made, new facilities will be built, and the Probation Department’s troubles will be over.

Closing facilities and transferring troubled youths to other buildings will not change the irresponsible management that is endangering and killing people.

May 14, 2023

This version of events is the unions’ self-serving fantasy, but it was sufficiently compelling that freshman Assemblymember Blanca Pacheco (D-Downey) sponsored the bill and three of the five L.A. County supervisors — Hilda Solis, Janice Hahn and Kathryn Barger — support it.

The actual story is a lot more troubling. It’s too long and aggravating to recount in full, but it’s important to know the recent highlights, beginning when probation officers refused to report to work, creating a staffing crisis at the juvenile halls.

In just its first month of operation since reopening, L.A. County’s Los Padrinos Juvenile Hall has seen a brawl and a chaotic escape attempt.

Aug. 27, 2023

Dishonesty was endemic. Probation workers who did come to work faked written certification that they had checked on juveniles in their rooms every 15 minutes as required by law. Their supervisors faked certification that they reviewed videos that were supposed to prove the checks took place. Inadequate staffing meant no daytime activities for juveniles except TV and video games, and nighttime lockdowns that compelled juveniles to relieve themselves in their rooms because they would not be let out to use toilets.

The Los Angeles County Probation Department has a history of sitting on state funds meant to be distributed to youth development programs, undermining its core purpose.

Aug. 6, 2023

There were several fentanyl overdoses, and one teenager died. There were fights and beatings. The two juvenile halls were finally stripped of their certification to operate, and the county reopened a shuttered third juvenile hall and transferred most of the young wards there. Then a gun was found inside, and there were a riot and an escape, and continuing failure of officers to report to work.

More money for better buildings won’t solve these problems. It’s true that Central Juvenile Hall, now finally closed for good, is more than a century old, but the problem was never the hardware — the buildings, the equipment, the vehicles. The problem is the software — the people and their willingness to do the jobs for which they are paid. And a Probation Department culture that makes crisis a daily routine.

Los Angeles County juvenile halls have been cited for unconstitutionally poor conditions for decades. It’s too late to save them.

April 14, 2023

A decade ago, L.A. County used a state grant to remake Camp Vernon Kilpatrick, formerly a grim barracks in the Malibu hills that was famous for its sports programs. With more than $50 million in state money, L.A. County turned the grounds into Campus Kilpatrick, an impressive modern facility with a “homelike environment” and a design intended to foster intensive therapy and mentoring.

The new approach was dubbed the “L.A. Model” of juvenile rehabilitation. But it never materialized because the department staff didn’t fully sign on to the program. The grant bought great buildings that were essentially failures.

The common denominator is the board’s micromanagement. Less meddling may well have been part of an effective prescription had it come forward in recent years.

May 2, 2023

AB 695 would do the same, providing grants for more revamped probation hardware without improving the software. It’s as if the Probation Department were an airline whose flight crews don’t show up for work, and their union proposes to solve the problem of canceled flights by getting the taxpayers to buy the airline more jets.

Supervisors Lindsey Horvath and Holly Mitchell, to their credit, oppose the bill. In a letter to the Senate Public Safety Committee, they correctly note that the Probation Department caseload has dropped so precipitously that only six of 19 juvenile halls and camps remain open. The county hasn’t sufficiently downsized the payroll to account for the diminished task, yet the department still can’t get enough people to come to work.

L.A. County does need a juvenile justice bill, but one that would authorize rehabilitation services within its new Department of Youth Development, so that it can lawfully take on work that the Probation Department has shown itself to be inept at handling. The fact that the supervisors have not yet found an author to carry such a bill raises a question as to whether they are serious about their “care first” vision of juvenile justice. If so, they should get themselves a real bill that would do the job and urge the Senate to reject AB 695.

Amazon Vendor Central is one of three ways brands can sell their products on Amazon. Like any of the selling options, there are pros and cons to consider when deciding how you want to structure your Amazon business.

Before we discuss our tips for seeing success on Vendor Central, let’s dive into the basics of the program to understand whether it’s the best option for your brand.

What is Amazon Vendor Central?

Amazon Vendor Central is a platform that allows sellers to sell their products directly to Amazon, who in turn will ship and sell the items themselves.

This model is a great fit for brands who are looking to reduce overhead costs by eliminating the need for in-house warehousing, shipping, and customer service. With Vendor Central, Amazon handles all of these steps in the process.

Amazon Vendor Central vs. Amazon Seller Central

The next logical question many brands ask is the difference between Amazon Vendor Central and Amazon Seller Central.

In short, the main difference between the two is that with Vendor Central you sell to Amazon, and with Seller Central you sell the product yourself on Amazon. In the chart below, you can see a detailed outline of the exact differences between these two options.

With Seller Central, you have two options — selling the product to the end consumer yourself (2P) or selling the product to a 3P Seller who then ships and sells the product to the consumer (3P).

Amazon Hybrid Strategy

If you’re having a tough time deciding whether Vendor Central or Seller Central is right for your brand, you’re not alone. Limited space in Amazon’s fulfillment centers, MAP violations, and suppressed buy box are just some of the reasons brands are looking for a different solution.

Enacting a hybrid selling model on Amazon is one way to best service your customers and maintain profitability.

With the Amazon hybrid selling approach, a brand maintains both a 1P relationship through a Vendor Central Account with Amazon, and a presence on the 3P marketplace platform through their own brand’s seller account or through an authorized 3P preferred reseller, such as Blue Wheel.

Tips for Success on Vendor Central

Now that we’ve established what Vendor Central is, we can dive into some specific tips for success on the platform to help grow your brand’s presence!

Optimize Your Listings

Catalog management is one of the first steps to take when looking to increase sales on Amazon, whether you’re 1P or 3P. By optimizing your titles, product descriptions, A+ content, and product images, you’ll increase not only SEO but also customer satisfaction — because there’s nothing worse than receiving a product that is different from the description.

But catalog management is also an extensive process of classifying, aggregating, and managing product-related data to ensure consistency and accuracy across all sales channels. It’s a complex process that results in a seamless customer experience.

Increase Advertising Efforts

It’s well-known by now that Amazon Advertising is one of the most effective ways to grow your sales on the marketplace. Appearing first on the organic SERP is great — until your competitors buy that ad space and appear ahead of you. In order to maintain authority for your search terms, you need to advertise.

By establishing a strong brand identity on the platform, you’ll increase your product’s visibility, stand out from your competitors, and attract new customers. Taking a data-driven approach to Amazon Advertising is key to see success.

Enforce Brand Protection

Lastly, make sure you are protecting your brand on Amazon. If you’ve enacted a hybrid strategy, this is still critically important!

Remove unauthorized sellers to maintain control of your brand. Additionally, enforce MAP (minimum advertised price) policies to ensure your authorized resellers aren’t undercutting you. This will improve your chances of getting the buy box back on your listings as well!

If you’re tired of navigating the world of Vendor Central, partner with Blue Wheel as your . .

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Banana-Nut Pancakes (dairy products complimentary).

Pancakes are an acquainted view on the morning meal table, however my faves are these banana- and also- nut- jam-packed flap jacks. Offer them warm with real syrup – grocery store pancake syrup is absolutely nothing greater than synthetically seasoned sugar syrup. While I have actually designed this as a dairy-free dish, it functions just as well with normal milk, so please attempt this dish whatever your lactose resistance degree.

Return: 12 to 14 pancakes.

Canola oil, for frying pan.
2 mugs entire wheat bread flour.
1 tsp cooking powder.
1/4 tsp cooking soft drink.
1/4 tsp great sea salt.
1 1/4 mugs plus 2 tbsps soy milk.
3/4 mug mashed bananas (regarding 2 big really ripe bananas).
3 tbsps canola oil.
1/2 mug (2 ounces) sliced pecans.

Pre-heat a frying pan or huge frying pan over tool warmth till a spray of water sprinkled externally instantly develops dance beads. Or pre-heat an electrical fry pan to 350 levels F. Using a folded up paper towel, gently clean the frying pan with oil. Pre-heat the stove to 200 levels F.

In a tool dish, blend the flour, cooking powder, cooking soft drink, and also saltuntil incorporated as well as make a well in the.

In an additional tool dish, blend all the soy milk, the bananas, eggs, as well as oil up until well integrated. Put right into the well as well as mix till smooth; there can be a couple of little swellings continuing to be.

Utilizing 1/4 mug batter for each pancake, put the batter onto the frying pan. Prepare up until the sides of the pancake show up completely dry as well as the bottom is gold brownish, regarding 3 mins. Turn the pancakes as well as chef up until the various other side is gold, regarding 2 mins.

Offer cozy.

Offer them warm with real maple syrup – grocery store pancake syrup is absolutely nothing even more than synthetically seasoned sugar syrup. Utilizing 1/4 mug batter for each pancake, put the batter onto the frying pan. Prepare up until the sides of the pancake show up completely dry and also the bottom is gold brownish, regarding 3 mins. This pancake batter acts in different ways from others: The top might not come to be covered with little bubbles, which generally show the pancake is prepared to transform. Turn the pancakes and also chef till the various other side is gold, regarding 2 mins.

Solar energy

Solar power is ending up being extra as well as a lot more preferred as the oil rates maintain boosting. Solar power is a wonderful option that will certainly conserve us cash as well as perhaps also conserve our planet.
Solar energy is understood to be among the cleanest renewable resources around today. Just how solar energy jobs is by photovoltaic or pv floor tiles. These floor tiles alter the sunshine right into a type of electrical energy that can be made use of to warm our houses as well as also aid our yards expand.
What takes place in significance is that the floor tiles are developed with items of adverse and also favorable silicon that is location under a slim item of glass. This present is passed to the location in which we desire to warmth via as inverter which transforms it to electric power for us to utilize in our houses. This solar power can be utilized simply like power for lights, tvs, and also of program warmth.
Lots of people in the United States are currently including solar energy to various locations of their houses, such as cellars, sun parlors, or their whole residence. These photovoltaic panels are not really difficult to mount yet might be costly according to exactly how huge the panels require to be and also exactly how they require to be set up.
Numerous states throughout the United States are currently offering tax obligation rewards to people and also business that want to hidden to solar energy. You can learn even more by chatting with the U.S. Department of Agriculture or the U.S. Department of Energy.
Numerous institutions throughout the nationwide are currently transforming to solar power. Numerous institutions have really high electrical expenses, due to the electrical energy that is being made use of on an everyday basis to light the course areas, maintain our kids cozy throughout the wintertime as well as cool throughout the summer season.

Solar power is coming to be much more and also extra preferred as the oil rates maintain raising. Solar power is a fantastic option that will certainly conserve us cash and also possibly also conserve our planet.
Just how solar power jobs is by solar floor tiles. Numerous colleges throughout the nationwide are currently transforming to solar power.

Grilled Pineapple with Kahlua Dulce de Leche
2 (14 ounce) canisters sweetened compressed milk
1/3 mug Kahlua
1/4 tsp vanilla remove
1/2 ripe pineapple, ideally gold
Vanilla gelato, as required

Preheat stove to 425 levels F.

Pour sweetened condensed milk compressed a 9-inch deep-dish pie plate and cover and also foil. Place pie plate in a toasting frying pan as well as include adequate warm water to pan to get to midway up side of pie plate.

Include Kahlua and also vanilla remove to cooled down caramelized milk, blend up until smooth, and also period with salt. (The dulce de leche might be made 2 days in advance as well as cooled, covered.).

Grill the pineapple wedges, cut-side down, up until simply charred, concerning 2 mins on each cut side. Separate the wedges amongst plates, placed an inside story of ice lotion following to each, and also drizzle with the dulce de leche.

Cut the pineapple lengthwise via fallen leaves, maintaining leaves connected, right into 4 wedges. Grill the pineapple wedges, cut-side down, up until simply charred, concerning 2 mins on each cut side. Separate the wedges amongst plates, placed an inside story of ice lotion following to each, as well as drizzle with the dulce de leche.

Men walk through a lush plantation between Ecuador ’s balmy Pacific coast and its majestic Andes, lopping hundreds of bunches of green bananas from groaning plants twice their height.

Workers haul the bunches to an assembly line, where the bananas are washed, weighed and plastered with stickers for European buyers. Owner Franklin Torres is monitoring all activity on a recent morning to make sure the fruit meets international beauty standards – and ever more important, is packed for shipment free of cocaine.

Torres is hypervigilant because Ecuador is increasingly at the confluence of two global trades: bananas and cocaine.

The South American country is the world’s largest exporter of bananas, shipping about 6.5 million metric tonnes (7.2 tons) a year by sea. It is also wedged between the world’s largest cocaine producers, Peru and Colombia, and drug traffickers find containers filled with bananas the perfect vehicle to smuggle their product.

Drug traffickers’ infiltration of the industry that is responsible for about 30 per cent of the world’s bananas has contributed to unprecedented violence across this once-peaceful nation. Shootings, homicides, kidnappings and extortions have become part of daily life, particularly in the Pacific port city and banana-shipping hub of Guayaquil.

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“This is everyone’s responsibility: the person who transports it, the person who buys it, the person who consumes it,” vendor Dalia Chang, 59, a lifelong resident of Guayaquil, said of the cocaine trade. “They all share responsibility. They have ruined our country.”

The country, which is not a major cocaine producer, was especially rattled when a presidential candidate known for his tough stance on organized crime and corruption – Fernado Villavicencio – was fatally shot at the end of an August 9 campaign rally. He had accused the Ecuadorean Los Choneros gang and its imprisoned leader, whom he linked to Mexico’s Sinaloa cartel, of threatening him and his campaign team days before the assassination.

In addition to its proximity to cocaine production, cartels from Mexico, Colombia and the Balkans have settled in Ecuador because it uses the US dollar and has weak laws and institutions, along with a network of long-established gangs like Los Choneros that are eager for work.

Authorities say Ecuador also gained prominence in the global cocaine trade after political changes in Colombia last decade. Coca bush fields in Colombia have been moving closer to the border with Ecuador due to the breakup of criminal groups after the 2016 demobilization of the rebel group Revolutionary Armed Forces of Colombia, better-known by their Spanish acronym FARC.

A record 2,304 tonnes of cocaine was manufactured in 2021 around the world, mostly in Colombia, Peru and Bolivia. That year, nearly a third of the cocaine seized by customs authorities in Western and Central Europe came from Ecuador, double the amount reported in 2018, according to a United Nations report citing data from the World Customs Organization. Large drug busts have become more frequent and within the past month, European authorities have made record-setting busts after inspecting containers carrying bananas from Ecuador.

Authorities on August 25 announced Spain’s biggest cocaine haul yet: 9.5 tonnes hidden among cardboard boxes of bananas from Ecuador in a refrigerated container. Dutch officials also made their country’s largest-ever cocaine seizure last month – nearly 8 tonnes – in a container of Ecuadorean bananas. Authorities in Greece and Italy also announced seizures of cocaine hidden in Ecuadorean bananas this year.

Bananas headed to Europe are boxed at plantations, loaded into trucks that take them to massive warehouses in and around Guayaquil and transferred to maritime containers driven to an area port.

Then the ships head northeast to the Panama Canal, cross to the Caribbean Sea, and go east across the Atlantic.

Knowingly or not, banana growers, exporters, shipping corporations, port operators, private security companies, customs agents, agriculture officials, police, and buyers offer opportunities that drug traffickers have exploited.

Some traffickers have created front companies to mimic legitimate banana exporters, while others have acquired legitimate businesses, including plantations. They have found companies willing to be complicit in trafficking. They also have paid off, threatened or kidnapped truck drivers and other workers to help get cocaine into shipments.

Other traffickers have corrupted or intimidated police, customs agents, security guards and port workers to assist with — or ignore — tampering with containers at the ports.

Drug trafficking has contributed to the number of violent deaths in Ecuador, which doubled from 2021 to 2022, when 4,600 died, the most ever recorded in a year. The country is on track to break the annual record again, with 3,568 violent deaths tallied in the first half of 2023.

In Guayaquil, where maritime shipping containers are part of the landscape, people live in fear these days. Pedestrians don’t dare take their phones out of their pockets. Convenience stores have floor-to-ceiling metal bars that prevent customers from entering from the sidewalk. Restaurants that survived the pandemic close early.

Along with the rise in homicides, the amount of cocaine seized at the country’s ports has increased, too, reaching 77.4 tonnes last year. That is more than three times the amount seized in 2020.

National Police General Pablo Ramírez, Ecuador’s national director of anti-drug investigations, attributed the change to increased smuggling, not better enforcement.

Police data also show that of last year’s total, a record 47.5 tonnes of cocaine were found in shipments of bananas, even though the fruit’s exports dropped 6.4 per cent, compared to 2021.

No more than 30 per cent of containers is currently inspected at Ecuadorean ports, a process done manually or with drug-sniffing dogs. President Guillermo Lasso’s government says it wants to use scanners on entire containers. Twelve of those machines were supposed to be operating already, but Ramírez said that has not happened yet.

Ramírez said he expects all ports to have operational scanners by mid-2024. He said two ports have tested the scanners to smooth out internal procedures and train the people who will be working with the machines.

The operator of the largest port in Guayaquil, Contecon Guayaquil SA, turned down Associated Press requests for an interview and access to the port to see existing security procedures. In response to written questions about the measures, spokeswoman Alexandra Pacheco said in a statement that the operator entered into an agreement with the National Police in 2022 to, among other things, “reinforce operations in the port”. She added that the operator plans to spend about US$15 million on the scanners.

Jose Hidalgo, executive director of the Association of Banana Exporters of Ecuador, said the industry faces greater exposure to trafficking than other commodity exports because of the volume of containers that it uses.

“It is because of bananas that there are so many ports,” Hidalgo said. “It opens routes to other export products.”

He explained that exporters spend about US$100 million annually on security measures, which include surveillance cameras at plantations, GPS monitoring of trucks, and the identification of land routes that require police patrols to keep criminals away.

Nonetheless, some exporters have been accused of being complicit or directly involved in trafficking cocaine.

Torres, the plantation owner, would like to see that type of exporter kicked out of the industry. But there is no regulation that can be used to revoke a company’s banana-exporting permission when the business is tied repeatedly to drug trafficking.

“It bothers me so much,” Torres said. “My people work with bananas, they don’t work with drugs. It’s a flagship product, the best in the world, and to see it tainted like that is unfortunate.”

In recent years, the safety and efficacy of AAV as a vector for gene therapy have been validated by clinical trials of AAV-mediated liver-targeted therapy for hemophilia A and B [36,37,38]; consequently, in vivo clinical applications of AAV-based vectors for liver-targeted gene therapy are rapidly increasing. Liver-targeted gene therapies for FD, namely FLT190 [39] and ST-920 [40], are both in phase 1/2 clinical research, and both have achieved good safety and efficacy. Therefore, we attempted to design a highly efficient AAV2/8-hGLA to establish a proof-of-concept for the clinical development of liver-targeted gene therapy for FD so as to improve the therapeutic effect and reduce the dose of AAV vector, thereby reducing the cost of gene therapy.

We validated the therapeutic effect of liver-targeted AAV2/8-hGLA in FD mice, which was produced using the HEK293 scientific research-scale manufacturing process. The results showed that α-Gal A activity significantly increased in FD mouse plasma and tissue. Indeed, the α-Gal A activity in the FD mouse livers was significantly higher than that in the FD group under the treatment of 0.75E + 12 vg/kg AAV2/8-hGLA (p = 0.0001), which resulted in a large amount of α-Gal A being secreted into plasma. The α-Gal A activity in the 0.75E + 12 vg/kg AAV2/8-hGLA–treated FD mouse plasma was approximately 53-fold higher than that of the FD group. The above results showed that AAV2/8-hGLA treatment results in high expression of α-Gal A in the livers of FD mice. The entire experiment was performed in the absence of immunosuppression, and following a single tail vein injection, the plasma enzyme activity in FD mice significantly increased in a dose-dependent manner, reaching a plateau in the second week and then maintaining sustained and stable plasma enzyme activity for 38 weeks.

After 12 weeks of AAV2/8-hGLA treatment, we found no significant difference in the plasma levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in FD mice treated with different doses of AAV2/8-hGLA (data not shown). The highest dose of AAV2/8-hGLA was 5E + 12 vg/kg, but no adverse reaction was observed in this group, confirming the safety of AAV2/8-hGLA. Compared with the FD group, at the dose of 0.75E + 12 vg/kg, the content and enzyme activity of α-Gal A in the tissues of FD mice were significantly increased in a dose-dependent manner. Reports in the literature show that as long as 5%–10% of α-Gal A enzyme activity remains, Gb3 and Lyso-Gb3 storage substrates can be effectively removed [41], indicating that the α-Gal A expressed in the AAV2/8-hGLA–treated FD mouse liver is effectively secreted into the plasma and taken up in FD mouse tissues, especially by the heart and kidney. Studies conducted earlier have shown that female FD mice exhibit reduced efficiency of transduction and transgene expression of AAV in comparison to males [25, 32, 39]. The effect may be caused by sex hormones, however, other studies pointed out that, unlike mice, androgens did not affect the transgene expression of AAV in nonhuman primates (NHPs) [42]. Thus, it is still necessary to determine if there exists a disparity in AAV2/8-hGLA transgenic expression between male and female FD mice. Although Gla KO mice have metabolomics profiles of FD, they do not exhibit the abnormal behavioral phenotype during our entire experiment cycle (from weeks 12 to 24). Despite this, a variety of preclinical pharmacokinetic and pharmacodynamic studies have been conducted on such models to evaluate enzyme replacement, substrate reduction, and gene therapy strategies for FD [43,44,45,46].

FD occurs as a result of the increased accumulation of Gb3 and Lyso-Gb3 in lysosomes, which leads to various complications [1]. Indeed, the storage content of substrates in patients is the current gold standard for the diagnosis and treatment of FD [47]. Our study showed that the content of Gb3 and Lyso-Gb3 in the plasma, urine, and tissues (heart, kidney, liver, and spleen) of FD mice treated with AAV2/8-hGLA was significantly reduced and tended to normalize compared with the FD group. Previous studies have shown that cardiac and renal failure are the most common and life-threatening complications in the end stage of most patients with FD disease; the main reason for the renal complications observed in FD is the poor clearance of Gb3, while the reason for the cardiac complications is the accumulation of Lyso-Gb3 [48]. Therefore, it is important to pay special attention to the clearance of renal Gb3 and cardiac Lyso-Gb3 when developing new therapies. Previous studies on FD have shown that poor clearance of substrate storage in the kidney is an important factor restricting the therapeutic effect on FD. However, our study showed that in FD mice treated with 0.75E + 12 vg/kg AAV2/8-hGLA, the residual Gb3 storage substrate in urine and kidney was < 10%, and the residual Lyso-Gb3 in heart was < 5%, indicating that AAV2/8-hGLA at a low dose had a good therapeutic effect in FD. And not only that the AAV2/8-hGLA treatment resulted in abundant α-Gal A presence in cardiomyocytes, renal tubular cells, and podocytes of FD mice. This could be taken as an indication that the Gb3 and Lyso-Gb3 could be efficiently cleared in these cells.

Notably, previous preclinical evaluations of AAV1- and AAV2-mediated gene therapy approaches in FD mice have shown suboptimal substrate clearance in key target tissues, particularly in the kidney [30,31,32]. Although the use of AAV8 vectors significantly improved transgene expression, normalization of renal Gb3 was only achieved when FD mice were treated at a young age (1 month old), before the onset of significant disease pathology [33]. The reason for this result may be related to the immune response, and the systemic expression of α-galactosidase using a constitutive promoter induces a strong immune response at low levels. The capsid, its genome, and transgene products are the main potentially immunogenic components of AAV vectors [49]. Other host-dependent and vector-dependent factors can modulate overall vector immunogenicity [49]. However, previous studies have shown that in the context of liver-directed gene transfer, transgene immunogenicity does not appear to be a problem compared with other tissues. Since the initial observation that mice expressing human factor IX in the liver are immune to transgenic products, several studies using AAV vectors in small and large animal models of genetic disease have shown that expression of the antigen in hepatocytes can promote strong antigen-specific immune tolerance [50,51,52]. Despite the success of AAV2/8-hGLA in clearing glycosphingolipids from peripheral organs, the enzyme generated is not expected to cross the blood–brain barrier (BBB). In the ensuing plan, we will develop a modified α-Gal A protein with a brain-targeting peptide in the future to achieve BBB transport and alleviate brain pathology and behavioral deficits [53].

Based on the results of previous studies, the infection efficiency and expression elements of the viral vector are the main factors affecting the therapeutic effect of gene therapy. In line with this, we aimed at improving the design of the transgene expression cassette to further enhance the AAV2/8-hGLA–mediated α-Gal A expression in plasma and tissues. As the GLA cDNA sequence used in AAV2/8-hGLA was optimized, further efforts have focused on engineering the AAV serotype to minimize the immune response elicited by the AAV vector. In vitro studies have demonstrated that AAV plasmid can effectively transfect liver cell and expression α-Gal A. In vivo studies have demonstrated that AAV2/8-hGLA can effectively transfect FD mouse livers, resulting in the expression and secretion of biologically active α-Gal A with high efficiency. Maximizing the efficacy of therapeutic AAV vectors is critical for clinical applications. Indeed, previous studies have shown that transduction of hepatocytes is more challenging in large animals, such as non-human primates (NHPs), compared with mice, with a 50- to 100-fold reduction in transduction efficiency [42, 54]. Furthermore, a more potent vector would allow to use lower vector doses, potentially allowing the vector to circumvent or minimize the anti-AAV capsid immune response associated with the administration of higher vector doses [54]. Notably, previous studies in transgenic mice overexpressing α-Gal A have demonstrated the safety of elevated systemic α-Gal A activity levels of up to 155- and 44-fold in the plasma and liver, respectively [55].

AAV2/8-hGLA (a safe and effective gene therapy for liver-specific expression) has several advantages over ERT (the current standard of care for FD): ① Liver-mediated AAV gene therapy has the potential to induce immune tolerance to the therapeutic target protein, while the formation of neutralizing antibodies against recombinases remains a limiting factor for the therapeutic effect of ERT. ② Gene therapy may become a “one-time treatment, life-long cure” treatment method for patients with FD, while ERT requires life-long treatment of patients, infusions of 2–4 h every 2 weeks, resulting in physical and psychological burdens for patients [19, 20, 56]. Furthermore, in mouse models of hemophilia B and Pompe disease, AAV-mediated gene therapy using preexisting antibodies against the respective therapeutic proteins resulted in sustained reduction or elimination of antibodies [43, 57, 58]. If true in humans, this would be a significant advantage, as it would allow the use of AAV gene therapy to treat patients with FD with preexisting α-Gal A antibodies (generated by ERT). The successful application of the AAV2/8 vector in FD mice has laid a theoretical foundation for the use of AAV2/8-mediated liver-specific expression gene therapy to treat LSDs. We will create a LSD gene therapy platform that we expect to reduce the dose and treatment cost of AAV vectors, thereby bringing hope for the realization of a one-time treatment for patients with LSDs.