Friday, March 22, 2024

New translation device aims to help TSA agents, travelers communicate at PHL - NBC Philadelphia - Translation

Going through the TSA checkpoint at the airport can be a stressful experience. Now, imagine that on top of that stress, you can't understand what the officers are saying to you.

Well, a new translation tool that fits in the palm of your hand is being tested at the Philadelphia International Airport with the hope of helping travelers who are not only limited speaking English but who are deaf or hard of hearing and those who are blind or have low vision.

Léelo en español aquí

"I meet people on a daily basis who do not speak English here at the airport," TSA officer Julio Rodriguez said as he explained why this new device is so critical.

A TSA officer or a traveler can speak into the device in their native tongue and it will translate that message into the selected language.

The device not only audibly reads the message out loud, but it also displays the message on its screen.

The hand-held unit on the left has translated “Please take off your shoes” in Spanish. The unit on the right has it translated into Vietnamese. (TSA photo)

These devices are part of a TSA pilot program being tested out at ten airports across the country, including PHL.

"We need to be able to communicate with our travelers," Christine Assili, of the TSA, said.

It's a device that's smaller than a smartphone, but it can translate more than 80 languages.

“Many times there’s miscommunication which of course when there is miscommunication there is misunderstanding and that’s the last thing we want for the traveling public," Assili explained.

The TSA says they've seen some challenges while testing them out including mistranslations that make them rephrase what they're trying to say, but they've also seen the benefit.

“I think with this device it will make it more convenient for us and easier for the passengers," TSA officer Youssef Sabiri told NBC10.

The devices are wireless and run off of WiFi or data connection. TSA officials say that the devices do not need to be connected to anything so they can be moved to wherever they're needed.

Last year, 28 million people traveled through the airport and 3.6 million of those landed on international flights.

“It’s a great place just to hear the energy and the languages...but also even better to be able to help folks along their journey," Denise Bailey, of the Philadelphia International Airport, said.

The devices join several other language access programs at PHL.

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Branched chemically modified poly(A) tails enhance the translation capacity of mRNA - Nature.com - Translation

Abstract

Although messenger RNA (mRNA) has proved effective as a vaccine, its potential as a general therapeutic modality is limited by its instability and low translation capacity. To increase the duration and level of protein expression from mRNA, we designed and synthesized topologically and chemically modified mRNAs with multiple synthetic poly(A) tails. Here we demonstrate that the optimized multitailed mRNA yielded ~4.7–19.5-fold higher luminescence signals than the control mRNA from 24 to 72 h post transfection in cellulo and 14 days detectable signal versus <7 days signal from the control in vivo. We further achieve efficient multiplexed genome editing of the clinically relevant genes Pcsk9 and Angptl3 in mouse liver at a minimal mRNA dosage. Taken together, these results provide a generalizable approach to synthesize capped branched mRNA with markedly enhanced translation capacity.

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Fig. 1: Conceptualization of chimeric mRNA bearing multiple poly(A) tails.
Fig. 2: Mechanistic characterization of mRNA stabilization by branched poly(A) tails.
Fig. 3: Multitail mRNA exhibited prolonged protein expression in vivo.
Fig. 4: Multiplexed genome editing in vivo using stabilized Cas9 mRNA.

Data availability

NGS data were deposited to the NCBI Sequence Read Archive database under the accession code PRJNA107297178. All data supporting the findings of the presented study are listed in the article and Supplementary Information is available upon reasonable request.

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Acknowledgements

We thank the MIT Department of Chemistry Instrument Facility for providing instrument access and N. Ye (MIT) for assistance on protein purification. We thank H. Zeng (Broad Institute) for his help in designing the STARmap/RIBOmap probes. We also thank other members of X.W.’s laboratory for helpful discussion throughout the project. X.W. acknowledges the support from the Searle Scholars Program, Thomas D. and Virginia W. Cabot Professorship, E. Scolnick Professorship, Ono Pharma Breakthrough Science Initiative Award, Merkin Institute Fellowship, and NIH DP2 New Innovator Award (1DP2GM146245-01). A.H. is a National Science Foundation Graduate Research Fellow. A.H. and D.R.L. were supported by NIH U01AI142756, R35GM118062, RM1HG009490 and the Howard Hughes Medical Institute (HHMI). This article is subject to HHMI’s Open Access to Publications policy. HHMI lab heads have previously granted a nonexclusive CC BY 4.0 license to the public and a sublicensable license to HHMI in their research articles. Pursuant to those licenses, the author-accepted manuscript of this article can be made freely available under a CC BY 4.0 license immediately upon publication.

Author information

Authors and Affiliations

Authors

Contributions

X.W. conceived the project. H.C. and X.W. designed experiments. H.C., D.L., J.G., A.A., Y.Z., J.T., J.R., A.H., F.K. and M.W. performed the experiments. H.C., J.G., S.L., A.H. and J.H. performed data analysis. X.W. supervised the work. H.C. and X.W. wrote the paper with input from all authors.

Corresponding author

Correspondence to Xiao Wang.

Ethics declarations

Competing interests

X.W., H.C., A.A. and J.G. are inventors on patent applications related to branched RNA. X.W. is a scientific cofounder, consultant and equity holder of Stellaromics and Convergence Therapeutics. D.R.L. is a cofounder, consultant and equity holder of Beam Therapeutics, Prime Medicine, Pairwise Plants, Chroma Medicine, Exo Therapeutics and Nvelop Therapeutics. The other authors declare no competing interests.

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Nature Biotechnology thanks the anonymous reviewers for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 Screening of oligonucleotide crosslinking chemistry.

(a) Summary of oligonucleotide chemical conjugation methods. Screening was performed using 15-nt poly-deoxyadenosine model substrates at micromolar concentrations. Modification handles were incorporated through oligonucleotide solid phase synthesis, followed by amine-NHS labeling and HPLC purification. (b) Gel electrophoresis of crude thiol-ene/yne oligonucleotide conjugation of 15-nt model substrates containing only one conjugation handle. (c) Gel electrophoresis of crude CuAAC and IEDDA 30-nt oligonucleotides bearing three alkyne/trans-cyclooctene handles reacting with 30-nt azide/tetrazine modified oligo. (d) Preliminary dual luciferase assay using branched mRNA prepared from CuAAC product mixture without HPLC purification using different equivalents of branched oligos versus stem oligo. Branching oligos contained modifications on the last six bases with either natural (5′ to 3′) or reversed (3′ to 5′) directionality. Time-course assay was performed as outlined in Fig. 1. n = 3 independent transfections for each construct. Mean ± s.e.m. P values were calculated by ordinary two-sided one-way ANOVA (comparison of means across time points). Gels are representative of at least two experiments.

Source data

Extended Data Fig. 2 Synthesis and characterization of topologically augmented branched mRNA.

(a-c) Representative HPLC purification and gel electrophoresis characterizations of branched oligonucleotides (oligo) containing 1-3 branching poly(A) tails. Fractions containing the desired products (boxed) were pooled and isolated. (d) Schematics of preparation of firefly luciferase (FLuc) mRNA constructs lacking poly(A) tail before ligation. FLuc mRNA without template encoded poly(A) tail was ligated to a scramble (non-polyA) stem oligo with 3′ end modifications (six phosphorothioate/2MOE at 3′ and terminal dideoxycytidine modifications) and internal alkyne (5-octadiynyl deoxyuridine or OU), with or without conjugation to 5′ azide labeled scramble or poly(A) branches with 3′ end modifications (the same as the stem oligo). (e) RNase H characterization of branched mRNA-oligo conjugates with branched poly(A) or scramble tails. The branching topology was confirmed by further band shift on TBU gel. (f) Branched poly(A) sequence, rather than chemical modifications alone, conferred enhanced protein expression over time. Relative FLuc luminescence was normalized to the scramble OU only oligo ligated mRNA at indicated time points. n = 3 independent transfections for each construct. Mean ± s.e.m. P values were calculated by two-sided unpaired t-test (with Welch’s correction) against the scramble OU + scramble azide construct at corresponding time points. (g) RNase H characterization of branched mRNA-oligo conjugates with multiple ploy(A) tails. mRNA with full-length hemoglobin UTRs and template encoded 100A-tail was ligated to 0 (mock ligation), 30 A, 60 A, or oligos with one, two, or three branched poly(A) tails and characterized by RNase H assay. The branching topology was confirmed by further band shift on TBU gel. Gels are representative of at least two experiments.

Source data

Extended Data Fig. 3 Dissecting the effects of branched poly(A) tails on mRNA stability and translation efficiency.

(a) Representative STARmap/RIBOmap images were acquired under the same confocal imaging settings from three independent experiments for each condition. STARmap versus RIBOmap characterization of different modified Firefly luciferase (FLuc) mRNA constructs and STARmap characterization of internal control Renilla luciferase (RLuc) mRNA were performed as outlined in Fig. 2. DAPI (blue), nuclei; FLuc amplicons (magenta); RLuc amplicons (yellow). Colocalized FLuc/RLuc amplicons in STARmap (white dots) were lipid transfection vesicles and were excluded from downstream quantification. Scale bar = 100 µm. (b-c) Violin plots of single-cell quantification of FLuc STARmap/RIBOmap amplicons. P values were calculated by ordinary two-sided two-way ANOVA. (d) Decay kinetics of internal control RLuc luminescence outlined in Fig. 2c normalized to luminescence at 8 hrs post transfection. n = 3 independent transfections. P values were calculated by ordinary two-sided two-way ANOVA (with Geisser-Greenhouse correction, decay across time points).

Source data

Extended Data Fig. 4 Branched poly(A) tails bind PABPC1 protein.

(a) Electrophoretic mobility shift assay of three branched poly(A) oligo with varying concentrations of GST-tagged recominant human PABPC1 protein (PABPC1-GST). The stem oligo was labeled with Alexa Fluore 546 at 5′end. Both stem and branching poly(A)’s were 30 nt and modified with PS-2MOE at the last six bases and ddC at the 3′ end. (b) Gel shift assay of the same modified poly(A) oligo without branching. Gels are representative of at least two experiments.

Source data

Extended Data Fig. 5 Chemical and topological augmentation stabilized poly(A) tails against nuclease degradation in vitro.

(a-d) Evaluation of chemically and topologically modified poly(A) tails in CAF1-CCR4 deadenylation assay. Four different Alexa-546 labeled poly(A) oligos were subjected to deadenylation using recombinant CAF1/CCR4 protein complex over the course of 110 min: linear unmodified 60 A oligo (a), end modified (six PS-2MOE at 3′ and terminal dideoxycytidine, the same set of modifications for all other oligos) 30 A oligo (b), end modified 30 A stem oligo + unmodified 30 A branches (c), and end modified stabilized 30 A stem oligo + end modified 30 A branches (d). (e-h) Evaluation of chemically and topologically modified poly(A) tails in HeLa cell lysate. Alexa-546 labeled poly(A) oligos (the same four constructs used in a-d) were subjected to digestion in HeLa cytosolic lysate for 220 min: linear unmodified 60 A oligo (e), end modified 30 A oligo (f), end modified 30 A stem oligo + unmodified 30 A branches (g), and end modified 30 A stem oligo + end modified 30 A branches (h). Gels were representative of two experiments.

Source data

Extended Data Fig. 6 Multi-tailed mRNA depends on the canonical eIF4-eIF3 translation initiation mechanism.

(a) Timeline of the knockdown (KD) experiments. HeLa cells were treated with siRNA cocktails targeting the corresponding eIFs 48 hrs post siRNA transfection, cells were reseeded and transfected with mRNA followed by protein quantification after 6 hrs. Successful knockdowns were confirmed by western blots and RT-qPCR at 48 hrs. (b,c,e) KD experiments for eIF4E/eIF4G/eIF3D. (d) Comparison of multi-tailed mRNA to regular mRNA in 4EGI-1 treated in vitro translation assay using rabbit reticulocyte lysate (RRL). Luciferase expressions were normalized to the wild-type condition for each construct. Mean± s.e.m. n = 3. P values were calculated by two-sided unpaired t-test for intra-construct comparison (KD vs WT) and by two-sided one-way ANOVA for cross-construct comparison. Western blots were representative of two experiments.

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Extended Data Fig. 7 Comparison of UTR optimized circRNA and multi-tailed mRNA using secreted NanoLuc reporter.

(a) Gel electrophoresis of circRNA encoding secreted Nanoluc (IL6-Nluc) generated through IVT, backsplicing, and enriched by RNase R treatment. Successful circularization was confirmed by RNase R resistance and slower mobility on TBU gel compared to corresponding intron-free linear RNA. (b) Comparison of UTR optimized circRNA and multi-tailed mRNA using IL6-NLuc 1 day after transfection with indicated amounts of RNA. Mock lig./mod-only/multi-tail mRNAs contained optimized UTRs, full m1ψ replacement and 100 A through IVT with mock ligation (mock lig.), ligated to modification-only oligo (mod-only), or modified multi-tailed oligo (multi-tail). CircRNAs were designed to contain optimized HRV IRES (with proximal loop Apt-eIF4G insertion) and 3′-PABP binding motif without addition of modified nucleotides. Cells were cultured in phenol red-free media. On each day, media was completely harvested and renewed with 150 μL fresh media and 5 μL of old media was diluted and used for luciferase assay. n = 3 independent transfections in each biological condition. Mean ± s.e.m. (c) IL6-Nluc signal over 14 days. n = 3 independent transfections in each biological condition. Mean ± s.e.m.

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Extended Data Fig. 8 Paired STARmap/RIBOmap effectively characterize the quantity and translatability of mRNA constructs delivered by LNP to murine liver.

(a) Schematics for evaluating two-gene STARmap/RIBOmap experiments in different lobes of murine liver. 2:1 NLuc/FLuc mRNA were co-encapsulated in LNP and administered by retro-orbital (R.O.) injection. 6 hrs post mRNA injection, the mouse liver was harvested in five lobes (caudate, left/right medial and lateral lobes). Each lobe was sections into two adjacent 10-μm slices, with one slice profiled by NLuc STARmap and FLuc STARmap and the other slice profiled by NLuc RIBOmap and Fluc STARmap. Whole sections were profiled in 291×291 μm2 tiles. (b, c) Quantification of numbers of NLuc and FLuc amplicons in each tile. n = 3379/3199 tiles. Slopes (NLuc/FLuc) were calculated by linear regression (least-square, Q = 5% for outlier rejection).

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Extended Data Fig. 9 Delivery of CRISPR editing using chemically and topologically modified mRNA.

(a) Comparison of Cas9 expression by western blot 48 hrs post transfection in HEK293T-uGFP cells. NLS, nuclear localization signal. Western blots were representative of two experiments. (b) FACS gating strategy for uGFP positive HEK293T cells. (c) Exemplary single-cell uGFP quantification for cells treated with unligated Cas9 mRNA only (Cas9-only) without sgRNAs, or with mock lig./mod-only/multi-tail Cas9 mRNA co-transfected with two sgRNAs. FACS were performed 72 hrs post mRNA transfection. n = 11845/10983/12334/12131 single cells. P value was calculated by two-sided one-way ANOVA. (d-h) Serum levels of Pcsk9 protein (d), Angptl3 protein (e), free cholesterol (f), total cholesterol (g), and triglyceride (h) over 4 weeks normalized to the Cas9-only group. Mean ± s.e.m. n = 4, biological replicates.

Source data

Supplementary information

Reporting Summary

Supplementary Table 1

Compiled sequences of genes/oligos/probes used in the study.

Source data

Source Data Figs. 1–4 and Extended Data Figs. 1–9

Compiled statistical source data for all figures presented.

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Chen, H., Liu, D., Guo, J. et al. Branched chemically modified poly(A) tails enhance the translation capacity of mRNA. Nat Biotechnol (2024). https://ift.tt/yUZ2p9t

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Thursday, March 21, 2024

Leeds lingo: Your ultimate urban dictionary for understanding Leeds student slang - The Tab - Dictionary

Your rundown of terms like Bakery, Monty B and RPP

With almost two semesters done and dusted, it’s probably fair to say that those of you that came to Leeds in September have about settled down.

You know where the libraries are – although you might still be plucking up the courage to enter Brotherton main circle – you’ve got the bus routes pretty much nailed down – that is, just the 56 – and you’ve probably found yourself a favourite coffee shop to go and study in – let me guess, it’s the Laidlaw or Business School Caffe Nero.

You’ve likely also started to come across, and maybe even begin using, some of the local Leeds Uni language: an ‘I’m going to Eddy B today’ here, and a ‘fancy a Bakery?’ there. However, for the freshers, and even us ancient final years, who are still left scratching heads at some of the terms flying around campus, we’ve compiled a handy guide to all the student lingo you need to know.

1. Eddy B

Let’s start nice and straightforward – Eddy B is short for Edward Boyle, one of the libraries on campus. It’s probably the most social out of all the libraries (and by social I mean loud) so best to avoid if you need to get any serious work done.

2. Town

Again, not too difficult, but if anyone says they’re going to town, they mean Leeds city centre. It’s just that ‘town’ makes people feel like they’re locals.

3. BNOC

A term that most people secretly wish could be used to describe them, BNOC stands for Big Name On Campus. In reality, it’s a bit of a pipe dream given there are tens of thousands of students in Leeds, but it’s still nice to aspire towards.

4. Monty B

Another B – this time to refer to the student halls, Montague Burton, near Woodhouse. We can kind of see the rationale behind this one, Montague Burton is a bit of a mouthful, after all.

5. Bakery

Technically, ‘Bakery’ does refer to a bakery, but more specifically it stands for Bakery164, the sandwich and coffee shop opposite the Parkinson Building. Getting your lunch here is not for the weak of heart because it’s always absolutely heaving, not just with students from Leeds Uni, but also those from the nearby sixth form college.

6. Dev

If there’s one thing Leeds students love, it’s giving accommodations unnecessarily short names. Here, Dev is short for Devonshire Hall, which is found just below James Baillie (JB – no, I’m not joking).

7. RPP

One that makes sense, RPP stands for Royal Park Pub, a student local in the middle of Hyde Park. Its £2 pints on Tuesdays and Friday karaoke nights have made it so that people will genuinely queue two hours in the cold for a chance to get in, and who can blame them.

8. Rah

It might have become an internet joke, but ‘rah’ is also definitely a term I’ve heard used seriously on campus. At this point, I’m not sure anyone knows exactly what it’s supposed to mean, but broadly speaking it’s a posh word used to express disbelief or excitability, depending on the situation.

9. Baccy

Look, I couldn’t not follow rah with baccy – the Hyde Park girlies have claimed it as their ironic (and sometimes unironic) tagline. For the non smokers out there, however, baccy is short for tobacco.

10. Brude

Another pub, Brude stands for Brudenell Social Club, a top tier establishment – I’m not biased – in the centre of Hyde Park. Aside from a good atmosphere, Brude also has a games room full of pool tables, perfect for a chill one with your friends, and a stacked line up of artists who’ll you find performing in one of its two event spaces every week.

11. Book club

Now someone could well be talking about an actual book club, but it’s more likely that they’re talking about Hyde Park Book Club, a favourite bar with Leeds students. It has a cosy candlelit main room for in the colder months and its outdoor seating becomes a sun trap in the summer, so if you’ve not already, we’d recommend checking it out.

12. Charlie Mo

Here is where I start to get caught out – Charlie Mo apparently refers to Charles Morris, one of the accommodations on campus. Maybe I’m getting too old but this is definitely a new one that’s started doing the rounds since my first year.

13. Henny P

What did I say – Leeds students are a sucker for an abbreviation. I could just about allow Charlie Mo, but Henny P for Henry Price? A line needs to be drawn somewhere and this might it.

Featured image via Google Maps

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Shohei Ohtani’s personal interpreter fired, accused of “massive theft” to pay off gambling debts - SoraNews24 - Translation

It’s a wild ending for one of the highest-profile pairings in the translation/interpretation world.

For the last few years, it’s been hard to think of a sweeter gig in the translation/interpretation field than Ippei Mizuhara’s job. Mizuhara has been in the business for a long time, but a key turning point in his career came in 2013, when he was working as an interpreter for a foreign player with Japanese professional baseball’s Hokkaido Nippon-Ham Fighters. That’s when Mizuhara happened to meet the Fighters’ highly anticipated rookie, a teenager named Shohei Ohtani.

Ohtani would become an instant all-star, and a few years later a Japanese Series champion. That success got him a ticket to head to the U.S. and play in Major League Baseball, and he took Mizuhara with him to serve as his personal interpreter. Shortly thereafter, Ohtani would become the most famous baseball player on the planet, the most accomplished two-way pitching/hitting phenomenon since Babe Ruth.

Having just signed a huge, long-term contract with the Los Angeles Dodgers, and constant endorsement offers coming in, Ohtani is financially set for life, and by extension so should Mizuhara have been, since Ohtani, like many Japanese pro athletes who achieve stardom overseas, prefers to speak through an interpreter for all of his interviews and public appearances. While there’s plenty of specialized jargon needed to discuss sports, it’s a less linguistically demanding topic than, say, high-end science or political debate. As an added perk, Mizuhara seemed to enjoy an easy-going friendship with Ohtani, if the atmosphere of their on-camera at-work interactions are anything to go by.

▼ Mizuhara interpreting for Ohtani at a game for the Anaheim Ducks, who play ice hockey right across the freeway from the freeway from the ballpark where Ohtani previously played for the Angels

But like with any profession, there are certain norms that have to be abided by in the interpretation field, and one of those is “Don’t steal millions of dollars of your client’s private funds to pay off your gambling debts.” Unfortunately, that’s what Mizuhara has been accused of doing, and he’s now reportedly been fired from his position as Ohtani’s interpreter.

According to multiple U.S. media outlets, Mizuhara became involved with an illegal southern California bookmaker named Mathew Bowyer. The interpreter racked up millions of dollars in gambling losses placing bets on soccer and other sporting events, though not baseball games, he asserts, saying “I never bet on baseball. That’s 100 percent. I knew that rule…We have a meeting about that in spring training.”

While Mizuhara’s gambling losses and the identity of who he owed money to aren’t in dispute, what happened next is murky. In speaking with American sports news network ESPN on Tuesday, Mizuhara said that since 2021, his gambling debt had grown to US$4.5 million. When Ohtani learned of this, Mizuhara said “Obviously, he wasn’t happy about it and said he would help me out to make sure I never do this again. He decided to pay it off for me.” A series of wire transfers were then made to Bowyer from Ohtani’s bank account, totaling at least one million dollars and described in the transaction documentation as “loans.” In the Tuesday interview, Mizuhara claimed that Ohtani himself made the transfers directly through his computer, not wanting to give the money to Mizuhara to pass off to Bowyer out of concern that the interpreter might gamble it away first.

However, the following day, when ESPN approached a representative for Ohtani for comments regarding Mizuhara’s story, the baseball player’s legal team instead refuted Mizuhara’s claim that Ohtani had personally made the transfers, or that he had even known about them, saying “In the course of responding to recent media inquiries, we discovered that Shohei has been the victim of a massive theft and we are turning the matter over to the authorities.” An attorney for Bowyer, who is currently under federal investigation, also denied any direct contact between Ohtani and the bookmaker, saying “Mathew Bowyer never met, spoke with, or texted, or had contact in any way with Shohei Ohtani.”

Following these revelations, Mizuhara once again spoke with ESPN on Wednesday, withdrawing several of his claims, and now says that Ohtani never knew about his gambling debts nor agreed to help repay them.

Mizuhara described his gambling issues as an addiction, and there’s arguably some sympathy to be had for someone who, desperate for money and being so close to such a wealthy individual who he also perhaps saw as a personal friend, figured it wouldn’t hurt to take a little, especially if he believed he was going to be able to pay it back before anyone noticed it was missing On the other hand, it seems like if you feel close enough to someone to pluck a million or more from their bank account, you’d also feel close enough to ask. Mizuhara’s annual salary was also somewhere in the range of US$300,000 to US$500,000, according to Yahoo! Sports, which seems like it’s well above the threshold of earning enough to be responsible for your own finances.

Mizuhara’s departure is a sudden one, as he made the trip to Seoul to interpret for Ohtani during the Dodgers’ season opener that took place just this week. Technically, he appears to have been employed by the Dodgers organization, as reports are that it’s the team who’s fired him, though he was clearly hand-picked for the position by Ohtani himself. Who’ll be taking over interpretation duties for the star remains to be seen, but odds are there are going to be some new protocols put in place to separate his interpreter from his bank account.

Sources: ESPN, Los Angeles Times, Yahoo! Sports
Top image: Pakutaso
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