Thanks so much and good luck with 18:6 👍

You can follow this link and fill out an application to learn more about my coaching program: mikecola.typeform.com/to/xOjnQtp2 Best - Mike

Thanks so much.. Try increasing your protein I think it would be a good idea. my brother is a vegetarian and my best friend is vegan they both increase their protein and definitely felt a difference, muscle mass and strength.

@@MikeCola thanks for the advice😊 I had a terrible ankle injury two years ago and was on crutches for 11months, it’s still not quite right, have struggled regaining muscle.. have ordered some hemp powder and will make mindful decisions to up protein through food also 🤞🏻

🙌🙌

Glad you enjoyed it

Autophagy is a cellular process that involves the degradation and recycling of cellular components. Measuring autophagy can be complex because it involves multiple steps and can vary across different cell types and conditions. Here are some common methods used to measure autophagy: 1. **LC3 Conversion and Detection:** - **LC3-I to LC3-II Conversion:** LC3 (microtubule-associated protein 1A/1B-light chain 3) is a key protein involved in autophagy. During autophagy, LC3-I is converted to LC3-II, which is associated with the autophagosome membrane. This conversion can be detected using Western blot analysis. An increase in the LC3-II/LC3-I ratio is often used as an indicator of autophagy. - **Immunofluorescence Microscopy:** The localization of LC3 can be visualized using fluorescence microscopy. The formation of LC3-positive puncta (small dots in the cytoplasm) indicates the presence of autophagosomes, which can be quantified to measure autophagy levels. 2. **p62/SQSTM1 Degradation:** - p62/SQSTM1 is a protein that binds to ubiquitinated proteins and is selectively degraded by autophagy. A decrease in p62 levels, detected by Western blot or immunofluorescence, is often used as a marker of increased autophagy. 3. **Autophagosome and Autolysosome Counting:** - **Electron Microscopy:** This is a gold-standard method for directly visualizing autophagosomes and autolysosomes at the ultrastructural level. Counting these structures provides a direct measure of autophagy. - **Fluorescence and Confocal Microscopy:** Cells can be stained with fluorescent markers specific to autophagosomes and lysosomes. The colocalization of these markers can be analyzed to measure autophagic flux. 4. **Lysosomal Degradation Activity:** - The degradation activity of lysosomes can be measured using substrates that release a fluorescent signal upon cleavage by lysosomal enzymes. An increase in lysosomal degradation activity can be indicative of autophagic flux. 5. **Autophagic Flux Assays:** - Autophagic flux refers to the complete process of autophagy, including the formation of autophagosomes, their fusion with lysosomes, and the degradation of cargo. To measure flux, inhibitors like bafilomycin A1 (which blocks the fusion of autophagosomes with lysosomes) are used in combination with the detection methods above. By comparing conditions with and without the inhibitor, researchers can determine the rate of autophagic flux. 6. **GFP-LC3 or mCherry-GFP-LC3 Reporters:** - Transfection of cells with GFP-LC3 or mCherry-GFP-LC3 constructs allows for real-time monitoring of autophagy. GFP-LC3 transitions from a diffuse pattern to punctate structures as autophagosomes form. The mCherry-GFP-LC3 tandem fluorescent reporter allows for the distinction between autophagosomes and autolysosomes, as the GFP signal is quenched in the acidic lysosomal environment, while the mCherry signal remains. Each method has its advantages and limitations, and often, multiple approaches are used together to get a comprehensive understanding of autophagy in a given system.

About 100 grams

@MikeCola ah so you measure your food on a scale

@@MikeCola too much!

The container that I buy it it is 142 g so I just take 2/3

A little wine now and then 🍷🙌

Thanks 🙌

Glad it was helpful!

@@MikeCola very!

You are so welcome

Much appreciated!

Yes, that can work 👍

So sorry about your husband. I'm glad you're finding my videos helpful and you're losing weight 👍 stay with it 🙌

Glad it was helpful!

Thanks so much. I will, change the music. Thanks for the tip.

Maybe not the music but he needs to change?

You are so welcome

Thanks 👍

You can do it 🙌

Nice 🙌

You're welcome!

As long as you're taking enough protein nutrition, I think it'll be OK... if you start getting weaker in the gym, then I would take a break

P.S olive oil, nuts , dark chocolate calories may add up... healthy yes, calorie dense also yes.

Sorry, Kirill, we had technical difficulties... the picture and sound was going in and out. I'll repeat this livestream

@@MikeCola I understand, don't worry!

Thanks so much 😄