Clock And Cycle: Wnt Signaling

The circadian clock controls daily life activities; the cell cycle is what allows cells to proliferate.  The two are linked via cell signaling, and in the intestines Wnt signaling is, unsurprisingly, important.  This has applications for modulation of the cell cycle for therapeutics.  Abstract:

Like two dancers, the circadian clock and cell cycle are biological oscillators engaged in bidirectional communication, resulting in circadian clock-gated cell division cycles in species ranging from cyanobacteria to mammals. The identified mechanisms for this phenomenon have expanded beyond intracellular molecular coupling components to include intercellular connections. However, detailed molecular mechanisms, dynamics, and physiological functions of the circadian clock and cell cycle as coupled oscillators remain largely unknown. In this review, we discuss current understanding of this connection in light of recent findings that have uncovered intercellular coupling between the circadian clock in Paneth cells and the cell cycle in intestinal stem cells via WNT signaling. This extends the impact of circadian rhythms regulating the timing of cell divisions beyond the intracellular domain of homogenous cell populations into dynamic, multicellular systems. In-depth understanding of the molecular links and dynamics of these two oscillators will identify potential targets and temporal regimens for effective chronotherapy.

Wnt Inhibitor And Small Cell Lung Cancer

A Wnt signaling inhibitor has some activity against small cell lung cancer.  Abstract:

Small cell lung cancer (SCLC) is the most aggressive type of lung cancer due to a fast tumor doubling time and early hematogenous spread. Advances in the treatment of non-small cell lung cancer using targeted therapies having been made, but no targeted drugs for SCLC have been approved. The Wnt signaling pathway is associated with tumor progression and metastasis; therefore, the inhibition of Wnt/β-catenin signaling is a strategy for anticancer drugs. Tankyrase 1 (TNKS1) is overexpressed in a number of types of cancer and XAV939 is a small molecule inhibitor of TNKS1 which may inhibit tumor growth. The present study aimed to investigate the potential molecular mechanisms underlying XAV939-induced suppression of the viability of SCLC cells. MTT assays were used to determine the viability-inhibition rate of cells and to identify the drug concentration which optimally inhibited cell viability. Flow cytometry was used to determine whether XAV939 induced apoptosis of SCLC cells, and to analyze the effect of the drug on the cell cycle. The results of the present study identified that XAV939 inhibited the viability of NCI-H446 cells in a dose-dependent manner, but cisplatin inhibited NCI-H446 cell viability in a time- and dose-dependent manner. The combination of XAV939 and cisplatin exhibited a slightly more pronounced inhibition of cell viability at an increased dose of XAV939. In addition, XAV939 markedly induced cell apoptosis of the SCLC cell line H446 by increasing the proportion of cells in the G0/G1 phase, leading to inhibition of the cell cycle. The results of the present study indicated that XAV939 inhibited the viability of the NCI-H446 SCLC cell line by inducing cell apoptosis through the Wnt signaling pathway. Therefore, XAV939 may be useful for the treatment of SCLC.

Mushroom soup

Hello, everyone!

Today's weekend cooking ended up being an easy mushroom soup. Why? Because I had made a salad of beans and corn that no one was eating and the mushrooms in the supermarket were on sale.

 
Mushroom soup

Ingredients:
1 sweet red pepper, cut into small cubes
1 15-oz can of corn (buy the one without added sugar)
1 15-oz black beans
1 medium size sweet onion, cut into small cubes
20-30 oz any mushrooms, washed and cut into small cubes
salt, turmeric, black pepper, cayenne pepper to taste
2 Tbsp olive oil
1-2 Tbsp wheat flour


Directions:
In a pan, heat 2 Tbsp of olive oil and add the mushrooms. Cover with a lid and let the mushrooms simmer in their own liquid. Stir from time to time; the mushrooms should be ready in about 10 minutes. In a soup pot, combined the pepper, onion, corn, beans and onion. Boil these in 4 cups of water. Once the onion is soft, add the flour and stir well. Add the mushrooms and more water to the desired density. Bring to boil and simmer for 10 minutes. Before turning off the heat, season with salt, turmeric, black pepper and cayenne pepper (optional).

Protein Glycosylation Targeting Immunotherapy

Another approach for anti-cancer immunotherapy, abstract:

Protein glycosylation provides proteomic diversity in regulating protein localization, stability, and activity; it remains largely unknown whether the sugar moiety contributes to immunosuppression. In the study of immune receptor glycosylation, we showed that EGF induces programmed death ligand 1 (PD-L1) and receptor programmed cell death protein 1 (PD-1) interaction, requiring β-1,3-N-acetylglucosaminyl transferase (B3GNT3) expression in triple-negative breast cancer. Downregulation of B3GNT3 enhances cytotoxic T cell-mediated anti-tumor immunity. A monoclonal antibody targeting glycosylated PD-L1 (gPD-L1) blocks PD-L1/PD-1 interaction and promotes PD-L1 internalization and degradation. In addition to immune reactivation, drug-conjugated gPD-L1 antibody induces a potent cell-killing effect as well as a bystander-killing effect on adjacent cancer cells lacking PD-L1 expression without any detectable toxicity. Our work suggests targeting protein glycosylation as a potential strategy to enhance immune checkpoint therapy.

Minoclonal Antibodies Against Clostridium Difficile

While probiotics may be a good choice for Clostridium difficile prevention, antibiotics may be required for infection.  Antibiotic resistance is becoming a problem.  Here is an interesting study on monoclonal antibody therapy for this difficult infection.  Abstract:

Background Clostridium difficile is the most common cause of infectious diarrhea in hospitalized patients. Recurrences are common after antibiotic therapy. Actoxumab and bezlotoxumab are human monoclonal antibodies against C. difficile toxins A and B, respectively. Methods We conducted two double-blind, randomized, placebo-controlled, phase 3 trials, MODIFY I and MODIFY II, involving 2655 adults receiving oral standard-of-care antibiotics for primary or recurrent C. difficile infection. Participants received an infusion of bezlotoxumab (10 mg per kilogram of body weight), actoxumab plus bezlotoxumab (10 mg per kilogram each), or placebo; actoxumab alone (10 mg per kilogram) was given in MODIFY I but discontinued after a planned interim analysis. The primary end point was recurrent infection (new episode after initial clinical cure) within 12 weeks after infusion in the modified intention-to-treat population. Results In both trials, the rate of recurrent C. difficile infection was significantly lower with bezlotoxumab alone than with placebo (MODIFY I: 17% [67 of 386] vs. 28% [109 of 395]; adjusted difference, -10.1 percentage points; 95% confidence interval [CI], -15.9 to -4.3; P<0.001; MODIFY II: 16% [62 of 395] vs. 26% [97 of 378]; adjusted difference, -9.9 percentage points; 95% CI, -15.5 to -4.3; P<0.001) and was significantly lower with actoxumab plus bezlotoxumab than with placebo (MODIFY I: 16% [61 of 383] vs. 28% [109 of 395]; adjusted difference, -11.6 percentage points; 95% CI, -17.4 to -5.9; P<0.001; MODIFY II: 15% [58 of 390] vs. 26% [97 of 378]; adjusted difference, -10.7 percentage points; 95% CI, -16.4 to -5.1; P<0.001). In prespecified subgroup analyses (combined data set), rates of recurrent infection were lower in both groups that received bezlotoxumab than in the placebo group in subpopulations at high risk for recurrent infection or for an adverse outcome. The rates of initial clinical cure were 80% with bezlotoxumab alone, 73% with actoxumab plus bezlotoxumab, and 80% with placebo; the rates of sustained cure (initial clinical cure without recurrent infection in 12 weeks) were 64%, 58%, and 54%, respectively. The rates of adverse events were similar among these groups; the most common events were diarrhea and nausea. Conclusions Among participants receiving antibiotic treatment for primary or recurrent C. difficile infection, bezlotoxumab was associated with a substantially lower rate of recurrent infection than placebo and had a safety profile similar to that of placebo. The addition of actoxumab did not improve efficacy. (Funded by Merck; MODIFY I and MODIFY II ClinicalTrials.gov numbers, NCT01241552 and NCT01513239 .).

Bezlotoxumab seems to be a promising therapeutic agent against this problem infection.

Modeling Chemical Communication

Metabolite-protein interactions can be used to model chemical communication in the body. Abstract:

Metabolite-protein interactions control a variety of cellular processes, thereby playing a major role in maintaining cellular homeostasis. Metabolites comprise the largest fraction of molecules in cells, but our knowledge of the metabolite-protein interactome lags behind our understanding of protein-protein or protein-DNA interactomes. Here, we present a chemoproteomic workflow for the systematic identification of metabolite-protein interactions directly in their native environment. The approach identified a network of known and novel interactions and binding sites in Escherichia coli, and we demonstrated the functional relevance of a number of newly identified interactions. Our data enabled identification of new enzyme-substrate relationships and cases of metabolite-induced remodeling of protein complexes. Our metabolite-protein interactome consists of 1,678 interactions and 7,345 putative binding sites. Our data reveal functional and structural principles of chemical communication, shed light on the prevalence and mechanisms of enzyme promiscuity, and enable extraction of quantitative parameters of metabolite binding on a proteome-wide scale.

Zesty salad for everyday

Hello, everyone!

I have been experimenting with my work lunches and decided that I needed as much spice and flavor as possible (to counteract the dullness of the work environment :)).

I love spicy-hot, check out this hot-hot walnut recipe. Therefore, I concocted a new salad mix that pleases my taste buds. I hope it pleases yours as well!
 
All major ingredients come from a can; therefore, the salad can be made in any season.

The salad is also fiber-rich, with no added sugar!


Zesty everyday salad

Ingredients
15.5-oz can corn kernels (make sure sugar is not added)
15.5-oz can black beans
20 oz pineapple chinks (no added sugar)
1 small sweet onion
hot pepper flakes (to taste)
hot sauce (to taste)
salt (to taste)


Directions
Dice the onion into small cubes (as small as possible). "Massage" (squeeze) the diced onion with some salt until the onion starts releasing its juice and it is less pungent. Drain the pineapple from its juice and cut the chunks into smaller pieces. Drain the beans and corn kernels, and combine these with the squeezed onion and pineapple. Mix with as much hot sauce and hot red pepper flakes as you like. Serve!


I keep my salad in a glass jar in the refrigerator for up to five days.

More On Tumor Immune Evasion

One mechanism of tumor immune evasion (at least in mice) is discussed here, abstract:

Conventional type 1 dendritic cells (cDC1) are critical for antitumor immunity, and their abundance within tumors is associated with immune-mediated rejection and the success of immunotherapy. Here, we show that cDC1 accumulation in mouse tumors often depends on natural killer (NK) cells that produce the cDC1 chemoattractants CCL5 and XCL1. Similarly, in human cancers, intratumoral CCL5, XCL1, and XCL2 transcripts closely correlate with gene signatures of both NK cells and cDC1 and are associated with increased overall patient survival. Notably, tumor production of prostaglandin E2 (PGE2) leads to evasion of the NK cell-cDC1 axis in part by impairing NK cell viability and chemokine production, as well as by causing downregulation of chemokine receptor expression in cDC1. Our findings reveal a cellular and molecular checkpoint for intratumoral cDC1 recruitment that is targeted by tumor-derived PGE2 for immune evasion and that could be exploited for cancer therapy.

Nervous System Immune Cells

There are distinct populations of central nervous system immune cells comparing states of health, aging, and disease Alzheimer's disease and multiple sclerosis). This may assist in devising new therapeutic strategies against those diseases as well as giving insights into the human aging process. Abstract:

Individual reports suggest that the central nervous system (CNS) contains multiple immune cell types with diverse roles in tissue homeostasis, immune defense, and neurological diseases. It has been challenging to map leukocytes across the entire brain, and in particular in pathology, where phenotypic changes and influx of blood-derived cells prevent a clear distinction between reactive leukocyte populations. Here, we applied high-dimensional single-cell mass and fluorescence cytometry, in parallel with genetic fate mapping systems, to identify, locate, and characterize multiple distinct immune populations within the mammalian CNS. Using this approach, we revealed that microglia, several subsets of border-associated macrophages and dendritic cells coexist in the CNS at steady state and exhibit disease-specific transformations in the immune microenvironment during aging and in models of Alzheimer's disease and multiple sclerosis. Together, these data and the described framework provide a resource for the study of disease mechanisms, potential biomarkers, and therapeutic targets in CNS disease. 

Gourmet cheese: make your own

Hello to all cheese lovers! I am one of you. 

Below is a recipe for all who would love to experiment with different flavors and herbs in their cheese.

 
Re-invented gourmet cheese

Ingredients
1:1 ratio of cream cheese and any cheddar cheese
1 Tbsp apple cider vinegar for every 8 oz of cheese (in this example recipe, I used 4 oz cream cheese and 4 oz cheddar cheese)
salt to taste
any herbs and spices of your choice: basil, hot pepper flakes, garlic powder, paprika, etc.

Directions
In a deep soup dish, combine all ingredients. Cover with another soup dish. For the amount of cheese indicated above (i.e., 8 oz), microwave for one minute. Carefully remove the dishes from the microwave and stir. Cover again with the second dish and microwave for another minute. Stir and pour on a piece of parchment paper. 
 

The spread of melted cheese on a piece of parchment paper

Smooth the melted cheese with a knife and sprinkle with herbs, if desired. Let it cool (you may need to refrigerate for faster cooling). Slice into pieces and serve.

I have tried variations of this cheese, including adding spoonfuls of crumbled feta after microwaving the cream cheese and cheddar. The result is delicious!

Let me know how you "customize" your cheese.

Bon appetit!  

By the way, the cheese is a wonderful addition to any frightfully cold winter day:

Icicles, as seen from my living room

Forever young, part II

Do you want to prevent premature aging and postpone for as long as possible today’s chronic diseases? Then you may try to modify your diet.  Max Lugevere’s YouTube channel is mostly dedicated to this topic. 

Max has also a website with more information on the topic.
 
Here are some of his YouTube videos:
 
Preventing Alzheimer's disease with FOOD

12 Easy and Powerful Holiday Health Hacks 

By watching some of these presentations, I learned that I should eliminate the carbohydrates from my breakfast. Why? Because carbohydrates spike up the insulin levels. Insulin, combined with the daily highest levels of cortisol (these are the levels when we wake up), “re-distributes” the weight from muscle to fat. I am still thinking how to reconcile this advice with my carbohydrate-rich breakfast!
 
For all of us, who want to be “forever” young and die in a nearly perfect condition from a painless cause, here is what Max recommends: Can We Reverse Aging?  The brief presentation is a good message, except for the statement that we can lengthen our telomeres, and therefore, reverse the aging process. In fact, the only thing we can accomplish in our normal cells is to not accelerate the telomere shortening (and therefore, speed up the aging process). 

Forget about telomere lengthening! Only cancer cells lengthen their telomeres.  But then, there is so much in common between the process of “keeping forever young” and cancer development! Think about it. Cancer cells are immortal; in the labs, we still grow cancer cells from individuals long dead.

By the way, if you are still unclear about what the telomeres are, I have previously written on this topic:

“Our genes are made of DNA, and this DNA forms the chromosomes in the cells ... Each of our chromosomes is protected at both ends with sequences called telomeres. As our cells duplicate (divide), the telomeres shorten. Shortening of the telomeres is equivalent to aging …, once the telomeres are of critical length, our cells stop dividing (this period is called senescence). However, changes (mutations) in a few genes may allow the cells to continue dividing. During such divisions, the chromosomes without their protective telomeres are subject to damage. The loss of chromosomal integrity may lead to the birth of cells with abnormal growth, and eventually, to cancer.”

Only our adult stem cells (which are normal cells), have an enzyme called telomerase. When functional, this enzyme counteracts to a certain extent the telomere shortening. All other normal cells seem to lack even this enzyme, therefore, the telomere length in such cells is not maintained.

There are certain conditions that may diminish the enzyme’s activity, and therefore, speed up the aging process. For instance, stress hormones slow down the activity of telomerase, making it more ineffective in preventing the telomere shortening. 

The fact that psychological stress shortens the telomeres faster than expected for a particular age was demonstrated with two groups of mothers: mothers of healthy children (low-stress group) and mothers of children with chronic illnesses (high-stress group). The study established that compared to the low-stress group, the high-stress group of mothers exhibited shortening of the telomeres that was equivalent to 9 – 17 additional years of age. Therefore, high stress may bring people closer to the catastrophic event when the chromosome integrity is endangered. 

Anyway, no normal cells can increase the length of its telomeres. At least, it has not been reported yet.

 To learn more about longevity, watch the lecture of Peter Attia, Reverse engineered approach to human longevity. 

Dr. Attia is taking a more reasonable approach to the dietary recommendations. When talking about healthy diets, he discusses the specific outcomes (biomarkers) we should aim at. He does not harp on us about any specific diets.

From all the confusing messages about diet/health I have heard, I could extract two conclusions:

1. If you already enjoy a healthy, balanced diet and your biomarkers are at the expected levels (see Attia’s talk), just go on with your current diet. If, however, you find it difficult to change your SAD (standard American diet), then try to limit your calorie intake or go on intermittent fasting (e.g., eat for eight hours a day and fast for 16 hours).

2. Do resistance training. I found this video. Probably, I may try the exercises soon.

By the way, the shortest description of a healthy diet is this: "Eat only the food that you great-grandma would have recognized as food". Well, this would exclude the majority of any supermarket offerings. And yes, it means that you need to start cooking.

Blood-Brain Barrier And Neurodegeneration

The blood-brain barrier is important for maintaining the health and proper function of the central nervous system, and this barrier breaks down in neurodegenerative disease.  Abstract:

The blood-brain barrier (BBB) is a continuous endothelial membrane within brain microvessels that has sealed cell-to-cell contacts and is sheathed by mural vascular cells and perivascular astrocyte end-feet. The BBB protects neurons from factors present in the systemic circulation and maintains the highly regulated CNS internal milieu, which is required for proper synaptic and neuronal functioning. BBB disruption allows influx into the brain of neurotoxic blood-derived debris, cells and microbial pathogens and is associated with inflammatory and immune responses, which can initiate multiple pathways of neurodegeneration. This Review discusses neuroimaging studies in the living human brain and post-mortem tissue as well as biomarker studies demonstrating BBB breakdown in Alzheimer disease, Parkinson disease, Huntington disease, amyotrophic lateral sclerosis, multiple sclerosis, HIV-1-associated dementia and chronic traumatic encephalopathy. The pathogenic mechanisms by which BBB breakdown leads to neuronal injury, synaptic dysfunction, loss of neuronal connectivity and neurodegeneration are described. The importance of a healthy BBB for therapeutic drug delivery and the adverse effects of disease-initiated, pathological BBB breakdown in relation to brain delivery of neuropharmaceuticals are briefly discussed. Finally, future directions, gaps in the field and opportunities to control the course of neurological diseases by targeting the BBB are presented.

Parkinson's Disease: Mitochondria Parked at the ER Hit the Snooze Button

A study evaluates the role of Parkinson’s disease genes in the sleep disturbances that accompany the disease.  Abstract:

Parkinson's disease patients report sleep disturbances well ahead of motor symptoms. In this issue of Neuron, Valadas et al. (2018) report that the disease genes pink1 and parkin exert novel, cell-type-specific effects to modulate ER-mitochondria contacts, neuropeptidergic transmission, and sleep patterns.