u/Murky-Commercial-112

Chill, you will be fine.

As a teenager and colleg kid, I used to fear things that mattered like starting a job or losing a job but that was every once in a while. Now I am an adult and I face decision making very often and I always avoid fearful outcomes and I end up missing out. I haven't reached out to regreting my lifestyle yet since I am still 30 yo but I know I will come down to regret wasting my life in fear. Can therapy or other things really help or is this a genetic thing that doesn't go away?

This is to understand the hiring freeze thing before following up with the PI, is it loosened for postdocs or still in place?

I had 2 3-year postdoc offers in Europe, I turned them down for a one-year position in the USA. Before criticizing me, I was under the impression that the USA is the land of opportunities and I should stay there and fight to achieve greatness, I was burned out though. Six months into my very temporarily role, I am already consumed because of the uncertainty, visa and immigration chaos. Though my current PI is very pleased with my work, their hands are tied because of funding. I just can't start the job application over again. I also don't have any emotional support in all of this. The more stable life I turned down in Europe is visiting me as a nightmare every night.

I cannot wait anywmore lol, these 45 business days for pp feels like eternity

Any recommendations for leases or property managments

Any recommendations for leases or property managment

Just wondered about how long should one pursue the proposed endeaovr after subitting I-140, I-485 and obtaining green card to maintain legality. I was thinking things could happen, funding get cut or get laid off etc and one may need to change or shift their research direction so how could we keep everyhing legal and transparent?

Hi everyone,

I received an endorsement from the Royal Society for the UK Global Talent visa (Exceptional Promise route), but due to personal and career circumstances I did not submit the actual visa application right away.

It has now been more than 5 months since the endorsement letter was issued. I am awarethat the visa application need to be submitted within 3 months of endorsement.

Has anyone been in a similar situation?

Were you still able to apply directly for the visa Or Did you need to request a new endorsement?

If you reapplied for endorsement, was the second process easier/faster?

Would appreciate hearing real experiences or advice.

Thanks!

I finished graduate school at 30 in the United States, but the journey has been far more difficult than I imagined. I moved to a new city for a contract job where I know no one, and I’ve been carrying the constant pressure of uncertainty ever since. My visa status needs renewal in six months, and my current job is only guaranteed for the same amount of time. At the same time, I’ve been pursuing a residency pathway, investing enormous amounts of money, time, and hope into the process, only to face endless silence and waiting with no clear outcome.

Living in this state of uncertainty has left me emotionally exhausted. Every part of my future feels unstable, my career, immigration status, finances, and even the possibility of starting a family someday. It’s difficult to make long-term plans when I don’t know what tomorrow will look like. I feel isolated, overwhelmed, and worn down by the constant need to survive and hold everything together while waiting for answers that never seem to come.

You guys have any updates on that?

For people working at Brookhaven National Lab (BNL), where do most people choose to live, and what’s the lifestyle realistically like?

I’m especially curious about:

• typical rent prices in different areas
• commute times and traffic
• whether a car is basically required
• best towns to live
• social life and things to do outside work
• safety and overall quality of life
• Is NYC nearby?
• do people feel isolated living near BNL or is there enough going on locally?

How do places like Patchogue, Port Jefferson, Stony Brook, Farmingville, Ronkonkoma, etc. compare in terms of affordability, commute convenienc and apartment availability

Would really appreciate honest experiences, especially from grad students, postdocs, or younger staff.

I started my current postdoc with the understanding that funding was only for 1 year becuase of funding limitation. During that time, I interviewed for another postdoc at a bigger lab with higher pay, a stronger university, amazing city and overall better long-term career prospects.

Recently, my current advisor secured funding to extend my position another year, but I’m still leaning toward the new opportunity. For those in academia, would you leave in this situation? And how would you handle the conversation with your current and prospective PI professionally ?

Anxiety and worry about visa, jobs, money and future is intense. Negative stories of faliures are all over the internet, this all could drive one crazy. How one should navigate this to stay sane and live their life?

There is a conference that I will be attending and will fly to and from chicago ohare airport. I am on OPT with expired F1 stamp. Will there be any problem flying or is it risky at all?

It makes me very anxious that it seems that EP accept everyone and they end up getting mostly RFE and denials. I applied with them and PP on March 31st. Has anyone got approved with this firm? It's just so stressful to watch everyone's critics of this firm and the number of denials they get

For a car this age and these miles, what are the main things that are likely to fail and should check before purchase?

I just want a car that last me couple years without headache and maybe resell it if plans changed

The buyer is asking for $4k

Hi all,

I’m running into an issue with substrate infiltration in Nicotiana benthamiana and would really appreciate any troubleshooting suggestions.

Setup:

• I transiently express my gene of interest via Agrobacterium infiltration.
• After ~4 days of expression, I infiltrate an exogenous substrate into the leaves.
• I then extract with ethyl acetate and analyze by GC-MS.

Problem:

• I cannot detect either the infiltrated substrate or the expected product in the extract.
• This is surprising because:
  • The reaction works well in crude protein extract (in vitro).
  • My extraction method seems fine, I can detect products derived from endogenous Nicotiana substrates using the same protocol.

Observations:

• The plants look somewhat weak/stressed after 4 days post-Agro infiltration.
• It seems like the issue is specifically with uptake or stability of the exogenous substrate in planta, not the enzyme or extraction method.

What I’ve considered so far:

• Poor substrate uptake through leaf tissue
• Substrate degradation or metabolism by the plant
• Volatility or loss during extraction
• Tissue damage affecting metabolism

Questions:

1. Has anyone successfully infiltrated small-molecule substrates into N. benthamiana and detected them reliably?
2. Could plant stress (4 dpi post-Agro) significantly reduce uptake or metabolic activity?
3. Any tips on improving substrate delivery? (e.g., solvent, surfactants like Silwet, concentration limits)
4. Could the substrate be getting rapidly metabolized or volatilized before extraction?

Any insights would be really helpful. Thanks!

Hi all,

I’m running into an issue with substrate infiltration in Nicotiana benthamiana and would really appreciate any troubleshooting suggestions.

Setup:

• I transiently express my gene of interest via Agrobacterium infiltration.
• After ~4 days of expression, I infiltrate an exogenous substrate into the leaves.
• I then extract with ethyl acetate and analyze by GC-MS.

Problem:

• I cannot detect either the infiltrated substrate or the expected product in the extract.
• This is surprising because:
  • The reaction works well in crude protein extract (in vitro).
  • My extraction method seems fine, I can detect products derived from endogenous Nicotiana substrates using the same protocol.

Observations:

• The plants look somewhat weak/stressed after 4 days post-Agro infiltration.
• It seems like the issue is specifically with uptake or stability of the exogenous substrate in planta, not the enzyme or extraction method.

What I’ve considered so far:

• Poor substrate uptake through leaf tissue
• Substrate degradation or metabolism by the plant
• Volatility or loss during extraction
• Tissue damage affecting metabolism

Questions:

1. Has anyone successfully infiltrated small-molecule substrates into N. benthamiana and detected them reliably?
2. Could plant stress (4 dpi post-Agro) significantly reduce uptake or metabolic activity?
3. Any tips on improving substrate delivery? (e.g., solvent, surfactants like Silwet, concentration limits)
4. Could the substrate be getting rapidly metabolized or volatilized before extraction?

Any insights or papers/protocols would be really helpful. Thanks!