A Novel Combination of Withaferin A and Sulforaphane have numerous anti-cancer effects

A Novel Combination of Withaferin A and Sulforaphane have numerous anti-cancer effects

http://www.bioportfolio.com/resources/pmarticle/1757328/A-Novel-Combination-of-Withaferin-A-and-Sulforaphane-Inhibits-Epigenetic-Machinery-Cellular.html?utm_source=dlvr.it&utm_medium=twitter

Excerpt: "With cancer often classified as a disease that has an important epigenetic component, natural compounds that have the ability to regulate the epigenome become ideal candidates for study. Humans have a complex diet, which illustrates the need to elucidate the mechanisms of interaction between these bioactive compounds in combination. 

 

The natural compounds withaferin A (WA), from the Indian winter cherry, and sulforaphane (SFN), from cruciferous vegetables, have numerous anti-cancer effects and some report their ability to regulate epigenetic processes. Our study is the first to investigate the combinatorial effects of low physiologically achievable concentrations of WA and SFN on breast cancer cell proliferation, histone deacetylase1 (HDAC1) and DNA methyltransferases (DNMTs). No adverse effects were observed on control cells at optimal concentrations. There was synergistic inhibition of cellular viability in MCF-7 cells and a greater induction of apoptosis with the combinatorial approach than with either compound administered alone in both MDA-MB-231 and MCF-7 cells. HDAC expression was down-regulated at multiple levels. Lastly, we determined the combined effects of these bioactive compounds on the pro-apoptotic BAX and anti-apoptotic BCL-2 and found decreases in BCL-2 and increases in BAX. 

Taken together, our findings demonstrate the ability of low concentrations of combinatorial WA and SFN to promote cancer cell death and regulate key epigenetic modifiers in human breast cancer cells."

My comment: "Serious science makes such discoveries almost daily when focusing on epigenetic regulatory mechanisms." Pseudoscientists claim that cancers are just bad luck. Most cancers are results from aberrant methylation patterns caused by oxidative stress, poor diet, bad life habits or viruses. This often leads to genetic mutations but they are not the actual reason for cancers. You can take care of your epigenome and impact your health, your childrens' health and your grandchildrens' health. Life is not driven by gene sequences, genes are driven by lifestyle. Don't get lost.

https://uknow.uky.edu/uk-healthcare/markey-cancer-center/plant-extract-traditional-indian-medicine-may-fight-blood-cancer

Bacteria have a sense of smell

Bacterial sense of smell - Incredible mechanism

https://www.technologynetworks.com/proteomics/news/scientists-investigate-bacterial-sense-of-smell-288948

Excerpt: "Scientists from MIPT, in collaboration with their colleagues from the Forschungszentrum Jülich, the Institut de Biologie Structurale (IBS) and European Synchrotron Radiation Facility (ESRF) in Grenoble, have proposed a universal mechanism for the “sense of smell” in bacteria. This was done by obtaining the structure of the NarQ protein from Escherichia coli (E. coli) – which belongs to a universal class of sensory histidine kinases that are responsible for transmitting signals to bacteria about their environment. The paper published in Science will help us understand how bacteria “communicate” with one another and form biofilms on sterile surfaces or inside the human body.

Drugs which affect bacteria’s “sense of smell” could potentially be used as substitutes for modern antibiotics. They do not kill bacteria; they simply give them signals that render them harmless to the human body. In theory, in this case it would be impossible for resistance to be developed.

The two components of a cell’s sense of smell

All cells are isolated from the environment by a dense membrane which virtually no chemicals are able to pass through. This enables the cell to keep its internal chemical conditions constant and function correctly. However, the membrane greatly limits the exchange of information with the environment. In order to find out what is happening on the outside, a cell uses special molecular machines – proteins. The proteins that are designed for communicating with the environment very often “live” in the membrane itself or close to it, and they are responsible for transmitting signals or chemicals into or out of the cell.

The most universal mechanism for bacteria to “sense” an environment are so-called two-component systems. Such systems consist of two proteins: a kinase, which receives the signal from outside of the cell and transmits it into the cell, and a response regulator, which receives the signal inside the cell and triggers subsequent reactions.

Molecular photography

A useful method of understanding how proteins function is to observe their structure at atomic-level accuracy. At present, most protein structures (more than 100,000) have been obtained using X-ray crystallography. This method involves observing the diffraction pattern of protein molecules ordered into a crystal lattice. However, this only gives the structure of one state of the protein, as in a photograph. If we can “photograph” the initial and final state of a process, we can make a guess as to how exactly the protein works when switching between these states.

Membrane “pistons” power a cell’s sense of smell

The authors of the study were able to obtain the structure of two states of the NarQ kinase from E. coli. This kinase “senses” the presence of nitrates in the environment and sends a corresponding signal through the cell membrane. As it turned out, the sensor in both of its two states is a dimer, i.e., two protein molecules work together to capture the nitrate. The first state is inactive – the protein is not bound to the nitrate ion and does not transmit a signal. The second state, on the other hand, is active, or signaling – in this state, the kinase transmits a signal into the cell to inform it that nitrates are present in the environment.

The protein structure in the active state was obtained for the most reliable “wild type” protein: without artificial mutations, which scientists often use to increase the stability of a protein. To obtain the structure in the inactive state, the authors mutated the site that the nitrate binds to. The stability of the protein was not affected; however, the nitrate no longer became bound to it, which gave the authors the opportunity to observe a kinase in a state of inactivity.

It was found that the signaling and inactive states differ only very slightly at the nitrate-binding site – by 0.5-1 angstroms, which is approximately one fifth of the size of the ion itself (1 angstrom is 10-10 meters). However, when this ion binds to the sensor, it causes huge changes in the protein: The helices of different monomers begin to move in different directions, like pistons. These “pistons” transmit the small change of 0.5-1 angstroms through the membrane, and their outer ends shift by approximately 2.5 angstroms in different directions. Inside the cell, in the HAMP domain, these shifts are converted into the rotation of two parts of NarQ relative to each other. Ultimately, the positions of the output helices change by as much as 7 angstroms, thus completing the signal transmission.

Aside from the structures in which the two proteins form a symmetrical pair, the scientists were able to produce a structure with an asymmetric position of the two proteins. In this state, the protein is “arranged” differently in the crystal and is bent strongly. However, the effect on the regulator remains virtually unchanged. This versatility of observed movement allows us to say that the signal transmission mechanism is universal and sensors of other chemical compounds, can operate via the same “piston-shift” mechanism.

“How signals are transmitted through the cell membrane is one of the most fundamental questions in modern biology. In this study, we showed in detail how a signal (in this case the binding of a nitrate) can be transmitted by hundreds of angstroms into the cells of bacteria and archaea, as well as fungi and plants. With a better understanding of the mechanisms of signal transmission, we can expect to learn how to manipulate such cells, and in particular, to try to weaken or neutralize the harmful effects of pathogenic microorganisms,” said Ivan Gushchin, the head of MIPT’s Laboratory of Structural Analysis and Engineering of Membrane Systems, commenting on the study." "

My comment: Evolutionists, which one evolved first, the kinase receptor or the response regulator? Do you think bacteria are using their sense of smell for randomly mutating their genes, or could there be a mechanism for genetic alterations? Of course there is. Rapid bacterial adaptation is based on clever signal transmission. This points to design and creation. Don't get lost.

Protein production needs perfect design

Engineering Protein Stability with Atomic Precision

https://www.technologynetworks.com/proteomics/news/engineering-protein-stability-with-atomic-precision-288982

Excerpt: "Proteins are the workhorses of biology. For example, they help convert light energy into sugar in plants, transport oxygen from our lungs to our muscles, and combine sugar and oxygen to release energy to make the muscles work. To perform these tasks, proteins must adopt specific 3D structures, called protein folds.

In chemical terms, proteins are polymers, or strings of amino acids, much like the beads of a necklace. There are 20 different chemistries of the amino-acid building blocks.

It is the combination of these along the protein string that determines how a protein folds up into its functional 3D shape. Despite decades of effort, scientists still don’t understand how biology achieves this protein-folding process, or, once folded, how protein structures are stabilised.

 

To address this problem, the Bristol team have combined two types of protein structure—called an a helix and a polyproline II helix—to make a stripped down, or simplified protein called a miniprotein.

This is basic science with the simple aim of seeing how small a stable protein structure can be. It is important, as natural proteins are usually very large and cumbersome structures, which are currently too complicated for chemists and biochemists to dissect and understand.

In the miniprotein, which the team call 'PPa', the two helices wrap around each other and their amino acids contact intimately in what are termed ‘knobs-into-holes’ interactions. This was expected, indeed the team designed PPa from scratch based on their understanding of these interactions.

Dr Emily Baker, who led the research in Professor Dek Woolfson’s laboratory, decided to change some of the amino acids in these knobs-into-holes interactions to non-natural amino acids, which the wonders of modern protein chemistry allow.

By doing this, Emily discovered that as well as the expected forces that hold proteins together, known as hydrophobic interactions, other more-subtle forces were at play in stabilising the miniprotein structure.

Chemists know these small forces as CH–p interactions, and they are found throughout the chemical world. When Drs Gail Bartlett and Kieran Hudson, also from the Bristol team, searched the thousands of natural protein structures available they found many examples of these CH–p interactions.

Moreover, the proteins that they occur in play roles in different biological process, many of which are associated with disease. This presents potential targets for new drugs, and the CH–p interactions may provide a valuable new route into developing these.

Dr Baker explained: "Our work has implications not only for understanding the basic science of protein folding and stability, but also for guiding the design and engineering of new proteins and drug molecules."

Professor Woolfson added: "This is precisely what the new Bristol BioDesign Institute is about. We aim to deliver the very best basic science. In this way, we will open unforeseen routes to translating fundamental science into biotechnology and biomedical applications."

My comment: Design of a 'simple' protein is an extremely challenging task for scientists. How come a random, unguided process could have been able to develop a single protein? Cellular parts and structures are made of perfectly designed proteins that also need to have been folded by atomic precision mechanisms by living cells. Life arises only from life itself. Random lottery is not able to produce complex proteins. It's not possible. That's why Intelligent Design and Creation. Don't get lost.

Checklist to confirm Biblical Creation and Intelligent Design

Checklist to confirm Biblical Creation and Intelligent Design - And to refute the theory of evolution

Things we are expected to observe whether Biblical creation and the global flood could be regarded as a potential theory for explaining the origin of life and the rich biodiversity. Here's the updated checklist.

 

1. No pre-cellular life forms have been observed. Check.
2. Unicellular life form has not been observed to evolve into multicellularity. Check.
3. Bacteria stay bacteria, dogs stay dogs etc. Check.
4. Large scale evolution has not been observed to happen. Check.
5. Organisms are able to rapidly adapt due to very sophisticated epigenetic mechanisms. Check.
6. Organisms may experience variation after adaptation. Check.
7. Variation modifies mating behaviour. Check.
8. Random genetic errors don't impact the biodiversity. Check.
9. There are a wide variety of efficient repair mechanisms in cells. Check.
10. A lot of rapidly buried organisms can be found in sediment layers. Check.
11. A lot of soft tissues can be found from bone samples from big animals like dinosaurs. Check.
12. Dinosaur bones still smell. Check.
13. There's an obvious lack of transitional fossils. Check.
14. Most fossils are exceptionally well preserved. Check.
15. Complex structures can be found from organisms in every fossil layer. Check.
16. The oldest human civilizations are only a few thousand years old. Check.

17. Human genome is rapidly deteriorating. There are ~200,000 disease-causing genetic defects in the DNA at population level. Over 10,000 new disease-causing mutations are discovered every year. 10% of people are living with a rare genetic disease.  Check.
18. A vast majority of human mutations have occurred during the last 5000 years. Check.

19. Scientists are in a hurry to develop gene editing techniques, such as CRISPR/Cas9. Check.

20. Numbers of mammalian chromosomes are only decreasing. Check.

21. There's still a missing link between humans and apes. Check.


There are a lot of reasons to believe in Biblical creation. Check.

Top six cancer fighting superfoods

Top six cancer fighting superfoods

http://www.health.com/health/gallery/0,,20430736,00.html

Excerpt: "To reduce your risk of cancer, look no further than your fridge. "All the studies on cancer and nutrition point to eating plant-based foods for their phytonutrients and other special compounds," says Richard Béliveau, PhD, chair in the prevention and treatment of cancer at the University of Québec at Montreal and author of Foods to Fight Cancer.

Aim for five to nine daily servings of all kinds of fruits and vegetables—especially these six superstars.

 

Tomatoes

This juicy fruit is the best dietary source of lycopene, a carotenoid that gives tomatoes their red hue, Béliveau says. And that's good news, because lycopene was found to stop endometrial cancer cell growth in a study in Nutrition and Cancer. Endometrial cancer causes nearly 8,000 deaths a year.

Helps fight: endometrial, lung, prostate, and stomach cancers

Your Rx: The biggest benefits come from cooked tomatoes (think pasta sauce!), since the heating process increases the amount of lycopene your body is able to absorb.

https://www.labroots.com/trending/cancer/6001/tomato-extracts-halt-stomach-cancer-cells

Broccoli

All cruciferous veggies (think cauliflower, cabbage, kale) contain cancer-fighting properties, but broccoli is the only one with a sizable amount of sulforaphane, a particularly potent compound that boosts the body's protective enzymes and flushes out cancer-causing chemicals, says Jed Fahey, ScD. A recent University of Michigan study on mice found that sulforaphane also targets cancer stem cells—those that aid in tumor growth.

Helps fight: breast, liver, lung, prostate, skin, stomach, and bladder cancers

Your Rx: The more broccoli, the better, research suggests—so add it wherever you can, from salads to omelets to the top of your pizza.

Berries

All berries are packed with cancer-fighting phytonutrients. But black raspberries, in particular, contain very high concentrations of phytochemicals called anthocyanins, which slow down the growth of premalignant cells and keep new blood vessels from forming (and potentially feeding a cancerous tumor), according to Gary D. Stoner, PhD, a professor of internal medicine at The Ohio State University College of Medicine.

Helps fight: colon, esophageal, oral, and skin cancers

Your Rx: Stoner uses a concentrated berry powder in his studies but says a half-cup serving of berries a day may help your health, too.

Walnuts

Their phytosterols (cholesterol-like molecules found in plants) have been shown to block estrogen receptors in breast cancer cells, possibly slowing the cells' growth, says Elaine Hardman, PhD, associate professor at Marshall University School of Medicine in Huntington, West Virginia.

Helps fight: breast and prostate cancers

Your Rx: Munching on an ounce of walnuts a day may yield the best benefits, Hardman's research found.

https://www.labroots.com/trending/cancer/6009/nutty-diet-fights-colon-cancer-return-study-suggests

Garlic

Phytochemicals in garlic have been found to halt the formation of nitrosamines, carcinogens formed in the stomach (and in the intestines, in certain conditions) when you consume nitrates, a common food preservative, Béliveau says. In fact, the Iowa Women's Health Study found that women
with the highest amounts of garlic in their diets had a 50 percent lower risk
of certain colon cancers than women who ate the least.

Helps fight: breast, colon, esophageal, and stomach cancers

Your Rx: Chop a clove of fresh, crushed garlic (crushing helps release beneficial enzymes), and sprinkle it into that lycopene-rich tomato sauce while it simmers.

Beans

A study out of Michigan State University found that black and navy beans significantly reduced colon cancer incidence in rats, in part because a diet rich in the legumes increased levels of the fatty acid butyrate, which in high concentrations has protective effects against cancer growth. Another study, in the journal Crop Science, found dried beans particularly effective in preventing breast cancer in rats.

Helps fight: breast and colon cancers

Your Rx: Add a serving—a half-cup—of legumes a few times a week (either from a can or dry beans that've been soaked and cooked) to your usual rotation of greens or other veggies.

My comment: Cancer is typically caused by aberrant methylation patterns resulted from poor diet and bad life habits. It's not only your epigenome that suffers due to weak nutrition, it's your offspring too. Take care of yourself.

Each of our cells has the same gene sequences

Traits are not determined by gene sequences - Epigenetic information layer is used for cellular differentiation

Each of our cells has the same gene sequences. However, there are hundreds of different cell types in our bodies. By which mechanism is a skin cell specialized into its own function? Why does a nerve cell have its own identity? Hundreds of different cell types are capable of producing hundreds of thousands of different proteins even though they have the same gene sequences. How is this possible?

There are several forms of biological information in our cells. Gene sequences constitute a digital body and a platform for other forms of biological information. For their successful differentiation, the cells need a specific information layer that functions on top of the genes. This kind of information is called epigenetic control of gene expression. If the epigenetic information layer is removed from the cell, it becomes a pluripotent stem cell that is in an open state to specialize in any task. Our genome has only about 19,600 protein encoding genes, but different proteins in our bodies are up to one million. Epigenetic control of gene expression and transcription makes this possible.

An epigenetic data structure consists mainly of chemical markers on top of genes. The most significant epigenetic marking of DNA is a methyl group (CH3) attached to a cytosine base. Also, methylation of DNA packaging proteins, i.e. histones, greatly affects the identity of the cell and the structure of the protein it produces. The number of different epigenetic markers on the DNA is not so high compared to RNA, whose epigenetic markers science has only started to understand in recent years. There are over a hundred different epigenetic markers of the RNA and they have a significant effect on protein production and embryonic development. Long and short non-coding RNA molecules form a significant group of epigenetic regulatory elements that participate in many vital regulatory tasks in our body.

The cell uses epigenetic information for many different functions. Genes can be suppressed, silenced or activated by methylating certain regions of them. The number of methyl groups in a gene and its different regions also plays a decisive role in the type and quality of the cell-produced protein. There are also genes in which only one addition or deletion of one methyl group has a significant influence on cell activity and identity. About 95% of plant genes are epigenetically suppressed, which explains their enormous potential for variation.

Perhaps the most interesting feature of epigenetic information is its analog nature. The team of McGill researchers, led by Professor Moshe Szyf of the Department of Pharmacology and Therapeutics, and Professor Ehab Abouheif, from the Department of Biology, found that the size of the Florida Carpenter Ants changed based on the amount of DNA methylation of a certain gene. Thus, the amount of methylation in the gene can act as a control knob, such as a volume regulator.

An example of epigenetic control of gene expression: Skin color.

If skin cells are programmed to produce white pigment instead of black, then which one is being changed, the gene sequences or the epigenome? The correct answer is: the epigenome.

This fact has been studied only by a few scientists, because it's too inconvenient fact for the theory of evolution. Here's one example:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2951359/

Unfortunately this group doesn't realize that point mutations don't modulate methylation patterns or levels on genes and histones. It's done by non coding RNA molecules.

Others have tried to find the reason for human skin color variation by examining sequence changes with poor results. They claim that light skin is determined by sequence changes in SLC45A2 gene, but do they tell that this claimed mutation is nearly absent in East Asia? So, they try to use another pseudoscientific explanation: Convergent evolution. This is the level of population genetics. It's false science.

There is no such a thing as mutation driven evolution or different human races. We all are created by loving God. Don't get misled.

European robin is able to maintain a quantum state longer than scientists in labs

European robin is able to maintain a quantum state longer than scientists in labs

https://www.newscientist.com/article/mg20927963-000-quantum-states-last-longer-in-birds-eyes/

Excerpt: "A process called the radical pair (RP) mechanism is believed to be behind the latter method. In this mechanism, light excites two electrons on one molecule and shunts one of them onto a second molecule. Although the two electrons are separated, their spins are linked through quantum entanglement.

The electrons eventually relax, destroying this quantum state. Before this happens, however, Earth’s magnetic field can alter the relative alignment of the electrons’ spins, which in turn alters the chemical properties of the molecules involved. A bird could then use the concentrations of chemicals at different points on its eye to deduce its orientation.

Intrigued by the idea that, if the RP mechanism is correct, a delicate quantum state can survive a busy place like the back of an eye, Erik Gauger of the University of Oxford and colleagues set out to find out how long the electrons remain entangled.

They turned to results from recent experiments on European robins, in which the captured birds were exposed to flip-flopping magnetic fields of different strengths during their migration season. The tests revealed that a magnetic field of 15 nanoTesla, less than one-thousandth the strength of Earth’s magnetic field, was enough to interfere with a bird’s sense of direction (Biophysical Journal, DOI: 10.1016/j.bpj.2008.11.072).

These oscillating magnetic fields will only disrupt the birds’ magnetic compass while the electrons remain entangled. As a weaker magnetic field takes longer to alter an electron’s spin, the team calculated that for such tiny fields to have such a strong impact on the birds’ compasses the electrons must remain entangled for at least 100 microseconds. Their work will appear in Physical Review Letters.

The longest-lived electrons in an artificial quantum system – a cage of carbon atoms with a nitrogen atom at its centre – survived for just 80 microseconds at comparable temperatures, the team points out. “Nature has, for whatever reason, been able to protect quantum coherence better than we can do with molecules that have been specially designed,” says team member Simon Benjamin of the Centre for Quantum Technologies in Singapore.

Thorsten Ritz of the University of California, Irvine, who helped perform the robin experiments, cautions that the RP mechanism has yet to be confirmed. But he is excited by the prospect of long-lived quantum states. “Maybe we can learn from nature how to mimic this,” he says."

My comment: Do they really think that an unguided process is able to develop such complex mechanisms that beat molecules designed by top scientists? European robin's quantum compass is an excellent example of Intelligent Design and Creation.

Epigenetic adaptation of stickleback to marine and freshwater conditions

Not random mutations but epigenetic adaptation of stickleback to marine and freshwater conditions

https://academic.oup.com/mbe/article-abstract/doi/10.1093/molbev/msx156/3813257/Genome-wide-DNA-methylation-profiling-reveals?redirectedFrom=fulltext

Excerpt from abstract: "The three-spined stickleback (Gasterosteus aculeatus) represents a convenient model to study microevolution - adaptation to a freshwater environment. While genetic adaptations to freshwater environments are well-studied, epigenetic adaptations have attracted little attention. In this work, we investigated the role of DNA methylation in the adaptation of the marine stickleback population to freshwater conditions. DNA methylation profiling was performed in marine and freshwater populations of sticklebacks, as well as in marine sticklebacks placed into a freshwater environment and freshwater sticklebacks placed into seawater. We showed that the DNA methylation profile after placing a marine stickleback into fresh water partially converged to that of a freshwater stickleback.
 

For six genes including ATP4A ion pump and NELL1, believed to be involved in skeletal ossification, we demonstrated similar changes in DNA methylation in both evolutionary and short-term adaptation. This suggested that an immediate epigenetic response to freshwater conditions can be maintained in freshwater population. Interestingly, we observed enhanced epigenetic plasticity in freshwater sticklebacks that may serve as a compensatory regulatory mechanism for the lack of genetic variation in the freshwater population.

For the first time, we demonstrated that genes encoding ion channels KCND3, CACNA1FB, ATP4A were differentially methylated between the marine and the freshwater populations. Other genes encoding ion channels were previously reported to be under selection in freshwater populations. Nevertheless, the genes that harbor genetic and epigenetic changes were not the same, suggesting that epigenetic adaptation is a complementary mechanism to selection of genetic variants favorable for freshwater environment."

My comment: Epigenetic adaptation is not a complementary mechanism for stickleback adaptations. All changes in organisms are based on these designed epigenetic mechanisms, not random mutations or selection. A cell without epigenetic information layers is an undifferentiated stem cell. There is no such a thing as mutation driven evolution. Don't get lost.

The most inconvenient findings that refute the theory of evolution

The most inconvenient findings that refute the theory of evolution

http://www.genesisalive.com/flood-artifacts.html

 

Nampa Image, 2 million charlie-years
Wright, Frederick, G. American Antiquarian 11:379-381 1889 and Scientific American, Nov. 9, 1889. An Image from Nampa, Idaho area was brought to the surface in a well drilling operation in 1889. The stone doll came from the 300-foot level of a well boring. “The record of the well shows that in reaching the stratum from which the image was brought up they had penetrated first about fifty feet of soil, then about fifteen feet of basalt, and afterwards passed through alternate beds of clay and quicksand down to a depth of about three hundred feet, when the sand pump began to bring up numerous clay balls, some of them more than two inches in diameter, densely coated with iron oxide." According to the United States Geological Survey the age of this strata "Plio-Pleistocene” of 2 million years ago using non-catastrophic measurements. It should be noted that the well log shows the image was found a bed of sand and far below a 15 foot lava layer of the Columbia basalts. These cover over 60,000 square miles of the Northwestern United States to a thickness measured in miles, which suggest they were laid down during the flood period. In this find we have evidence that a civilization existed before the catastrophe that covered the idol, confounding current Geological and Archeological origin's models but fully support a Biblical one.

  

Table Mountain CA, Gold mine finds, 30 million charlie-years
Wright, Frederick G. Science 20:275-27 1892
In February 1866, a skull was removed by one Mr. Mattison, in Calaveras County from a layer of gravel 130 feet below the surface. Later a human jaw, also inspected by J. D. Whitney, State geologist at that time who researched its location and authenticity. The jaw was given to a Dr. Snell by miners, who stated it came from the gravels beneath the lava cap at Table Mountain in Tuolumne County, CA. Noted geologists Mr. George H. Baker, Mr. King along with Professor Marsh, Professor Putnam and W. H. Dall attested to the fact that the Calaveras skull was found in place beneath a stream of gravel in the Table beneath this same stream of lava. Their position indicates they were over 30 million years old.

 

Nail in "Ancient" Sandstone, 408 million charlie-years
Brewster, David: Report of the British Association pt. 2, 51 1844
In a Scotland, at a sandstone quarry a report was made that a nail was found set in a block of sandstone. The relic was found in stone dated 360 and 408 million years old. The principal who made the find, David Brewster was a famous Scottish physicist and founder of the British Association for the Advancement of Science. Brewster stated: “The stone in Kingoodie quarry consists of alternate layers of hard stone and a soft clayey substance called ‘till’; the courses of stone vary from six inches to upwards of six feet in thickness. The particular block in which the nail was found, was nine inches thick, and in proceeding to clear the rough block for dressing, the point of the nail was found projecting about half an inch (quite eaten with rust) into the ‘till,’ the rest of the nail lying along the surface of the stone to within an inch of the head, which went right down into the body of the stone.”
  

Iron pot found in coal, 300 million charlie-years
Cremo, Michael A.; Thompson, Richard L. Forbidden Archealogy 2011. P. 806
Robert Nordling states; "While I was working in the Municipal Electric Plant in Thomas, Okla. in 1912, I came upon a solid chunk of coal which was too large to use. I broke it with a sledge hammer. This iron pot fell from the center, leaving the impression or mold of the pot in the piece of coal. Jim Stall (an employee of the company) witnessed the breaking of the coal, and saw the pot fall out. I traced the source of the coal, and found that it came from the Wilburton, Oklahoma, Mines.” Wilburton area coal is said to be over 300 million years old using uniform geologic tables. 

  

Shoes in Stone, 200 million charlie-years
Cremo, Michael A.; Thompson, Richard L. Forbidden Archealogy 2011. p. 807-8
“Mystery of the Petrified ‘Shoe Sole’ 5,000,000 Years Old,” by Dr. W. H. Ballou. DR. Ballou wrote: “Some time ago, while he was prospecting for fossils in Nevada, John T. Reid, a distinguished mining engineer and geologist, stopped suddenly and looked down in utter bewilderment and amazement at a rock near his feet. For there, a part of the rock itself, was what seemed to be a human footprint! Closer inspection showed that it was not a mark of a naked foot, but was, apparently, a shoe sole which had been turned into stone". The Triassic rock bearing the fossil shoe sole is now recognized as being more than 200 million years old. Mr. Reid took the specimen to the American Museum of Natural History where the expert staff seemed amazed, a report was written but when inquired of later it could not be found. The sole imprint was so clear a wear spot was visible a the heal pressure point. The artifact has been left a mystery to secular scientists, but not to those who hold the Bible as authoritative.
 

Sole print including trilobites, 500 million charlie-years
Corless William R. Handbook of Geological Enigma's Sourcebook Project 1980 p. 642
William Meister was collecting trilobites near Antelope Springs, Utah. Upon breaking open a clast rock the two parts fell open and there before his eyes were the impression of shoe soles impregnated with trilobites, an animal dated back 500 million years! Geologist after geologist has studied the imprints and rejected them due to the geologic location of trilobite's, which contradict the evolution chart of origins. One geologist familiar with the area visited the site and reported that surface rocks existed of the same nature of the one found, so Meister was exonerated from any kind of fraud. One geologist admitted that he "could not accept it", and added that no geologist would.

Fossil human-like footprints.
Corless Handbook of Geological Enigma's Sourcebook Project 1980 p. 668-9
Professor W. G. Burroughs, head of the department of geology at Berea College in Berea, Kentucky, reported in 1938 that he had identified a number of prints in the vicinity for years. Human and human mixed with animal prints have been known to be found in Australia, Dakota, El Salvador, Nicaragua, Massachusetts , Nevada, S Louis, Mo. Texas and Turkey.

Heidelberg Mandible
Dr. Otto Schoetensack, lecturer on Geology in the University of Heidelberg, often visited a fossil rich deposit in a gravel pit near the institution. After waiting and searching for twenty years, the owner of the pit, Herr J. Rosch, was able to inform him on October 21st, 1907, that his twenty years search had at last been realized. "Yesterday" he wrote, "the desired evidence was obtained, for 20 m. below the surface soil, and above the floor of my sand-pit, there was found the lower jaw of primitive man, in good preservation, and with all its teeth." Concerning the authenticity of the find there cannot be any doubt; the bed in which the mandible was covered by a series of deposits, amounting in all to 78 feet, over the mandible Dr. Schoetensack recognized twenty four different strata containing. They fall into three series - the uppermost, formed by recent loess a fine earth, a product of floods and drought; (2) the ancient loess a sandy loam, also a deposit from muddy waters; (3) the Mauer sands. In one of the lower strata of this series the mandible was found. In the lower strata, remains of the following extinct animals were found: " the lion, an extinct form of cat, a dog, two forms of bear, a species of bison, an early Pleistocene form of horse, and an early form of rhinoceros… "These are not primitive or simian characters, but the reverse; they are modifications confined, so far as we have yet discovered, to this peculiar variety or species of man "

"No revelation of prehistoric man could be more convincing than the discovery of the Heidelberg mandible. We have no shadow of doubt as to its authenticity or significance." So states Arthur Kieth in The Antiquity of Man. 1915. 233-4
 

Galley Hill, England
Keith, Arthur. The Antiquity of Man. 1915.
In 1888, workmen removing deposits at Galley Hill, near London, England, an exposed a fully modern human remains above a bed of chalk. The overlying layers were about 10 or 11 feet thick. Here was discovered a human skeleton firmly embedded in these deposits about 8 feet below the surface. A qualified observer determined this locale was not a burial site: “No doubt could possibly arise to the observation of an ordinary intelligent person of their deposition contemporaneously with that of the gravel. This undisturbed state of the stratum was so palpable to the workman that he said, ‘The man or animal was not buried by anybody.’” Numerous tools were also recovered from the Galley Hill site and surrounding area.

Clichy Skeleton In 1868
Cremo, Michael A.; Thompson, Richard L. (2011-01-30). The Hidden History of the Human Race
As documented to the Anthropological Society of Paris , one Eugene Bertrand claimed that he found parts of a human skull, along with a femur, tibia, and some foot bones, in a quarry near Avenue de Clichy. The bones were found 16+ feet beneath the surface. This layer was the same age in Geology terms as the Galley Hill skeleton was discovered. This would make the Clichy bones approximately 330,000 years old.

La Denise Skull Fragments, 2 million charlie-years 
La Denise, France In the 1840’s, pieces of human bone were discovered in the midst of volcanic strata, including a forehead section. Sir Arthur Keith wrote; “differs in no essential particular from the frontal bone of a modern skull.” The remains were found between two layers of lava. The first lava layer was from the Pliocene and the last from the Late Pleistocene, or up to 2 million years old. From Cremo, Michael A.; Thompson, Richard L. (2011-01-30). The Hidden History of the Human Race

 

Buenos Aires Skull. 1 million charlie-years

Cremo, Michael A.; Thompson, Richard L. (2011-01-30). The Hidden History of the Human Race.
A very strong case for anatomically modern humans existing in very early times comes from Argentina. In 1896, workers excavating a dry dock in Buenos Aires uncovered a skull in a layer referred to as “the upper-most portion of the Pre-Ensenadean stratum.” Placing the remains over 1 million years old as measured by evolution based timelines, far to old for the existence of modern man.

 

Foxhall Jaw, 2 million charlie-years
From Cremo, Michael A.; Thompson, Richard L. (2011-01-30). The Hidden History of the Human Race. In 1855, a human jaw was discovered at Foxhall, England, by workers digging in a quarry 16 feet below the surface. Collyer determined that this specimen was; “the oldest relic of the human animal in existence.” Artifacts were also uncovered in the same strata, over 2 million years dated by uniform means. Collyer was ecstatic about this find and presented it to the various authorities of the day including Thomas Huxley, Richard Owen and Charles Lyell. Huxley related that in his opinion the piece; “did not indicate it belonged to an extinct or aberrant race of mankind.” Others were skeptical. 

 

The Castenedolo finds, 4 million charlie-years
Keith, Arthur. The Antiquity of Man. 1915. Chap XIII
Late in 1860, one Giuseppe Ragazzoni, a geologist at Brescia, visited the area of Castenedolo, “Searching along a bank of coral for shells, there came into my hand the top portion of a cranium, completely filled with pieces of coral cemented with the blue-green clay characteristic of that formation. Astonished, I continued the search, and in addition to the top portion of the cranium I found other bones of the thorax and limbs, which quite apparently belonged to an individual of the human species.” Later he recovered more "modern" human fossils, to the astonishment of the world, including a man, women and two children. This finding was suppressed and has found little publicity as the layers, all water deposited, dated over 4 MY as determined by uniform geologic means. Giuseppe Sergi, famous anatomist of Rome, visited Ragazzoni in 1883 and verified the remains of the four individuals, an adult male, female, and two children. A noted authority of the day, Sergi believed the Castenedolo skeletons were authentic. As the skeptical reactions flowed in from others, he later said: “The tendency to reject, by reason of theoretical preconceptions, any discoveries that can demonstrate a human presence in the Tertiary is, I believe, a kind of scientific prejudice. Natural science should be stripped of this prejudice.” Later Sergi wrote: “By means of a despotic scientific prejudice, call it what you will, every discovery of human remains in the Pliocene has been discredited.”

The Honest end of Castenedolo
From Macalister’s Textbook of European Archaeology, 1921. Cambridge University Press.
Macalister mentions the Castenedolo finds “whatever we may think of them, have to be treated seriously.” Further he comments;“unearthed by a competent geologist, Ragazzoni and examined by a competent anatomist, Sergi.” He unknowingly predicts; “there must be something wrong somewhere.” “Now, if they really belonged to the stratum in which they were found,”…“this would imply an extraordinarily long standstill for evolution. It is much more likely that there is something amiss with the observations.” Finally and honestly asserting: “The acceptance of a Pliocene date for the Castenedolo skeletons would create so many insoluble problems that we can hardly hesitate in choosing between the alternatives of adopting or rejecting their authenticity.”

Savona Italy, Skeleton, 3 million charlie-years
Around 1850 while excavating for a Church in Savona, Italy, a fully modern human skeleton was unearthed 10 feet below the grade in a layer thought to be “millions” of years in geologic age -(geologists of the day put the age the layer over 3 million years old, again assuming uniform evolutionary geology was correct). The presenter of the find, Arthur Issel observed; “The body was discovered in an outstretched position, with the arms extending forward, the head slightly bent forward and down, the body very much elevated relative to the legs, like a man in the water." Animal bones were found scattered with the human remains in the same layer. As this skeleton appeared to be washed in by water, face down and trapped against the side of rock, this condition supports the find as but none other than a remnant the the great flood, 75 miles inland.

Human skull found at Stanford, Menlo Park, CA, 3 million charlie-years
Science, 69: suplii, Feb, 1 1929, from Corliss, William R. Handbook of Geological Enigma's Sourcebook Project 1980 p. 673
Discovering mastodon remains 22 feet below ground level near the campus, the observing scientists recalled a little reported earlier find of a modern human skull found nearby at same depth in the same strata. Three researchers are quoted discussing the implications. One, a Dr. J.W. Gidley is quoted as stating, "If this mastodon is of the late Miocene are early Pliocene as D. Blackwelder says it is, that sets it (the skull) back some two or three million years. And we have no evidence man has been here that long."

Switzerland, A human skeleton, 39 million charlie-years 
According to Gabriel de Mortillet, M. Quiquerez reported the discovery of a skeleton at Delémont in Switzerland in clays (water deposits) in the Eocene time – at least 39 million years old on the Geologic chart.

US Coal Man, 280 million charlie-years
Cremo, Michael A.; Thompson, Richard L. Forbidden Archaeology Torchlight Press 2011 p. 454
In December of 1862, The Geologist reported: “In Macoupin County, Illinois, the bones of a man were recently found on a coal-bed capped with two feet of slate rock, ninety feet below the surface of the earth. The bones, when found, were covered with a crust or coating of hard glossy matter, as black as coal itself, but when scraped away left the bones white and natural.” The coal in which the Macoupin County skeleton was a layer claimed to be over 280 million years

Reck’s Skeleton
In 1913, Professor Hans Reck of Berlin University, conducted investigations in the now famous graveyard at Olduvai Gorge in Tanzania. Seeing a protrusion, one of his companions noticed portions of a skeleton embedded in the rock. The workers labored to extract the remains out of the stone with hammers and chisels. This fully modern human was located in a layer far too "old". by evolution standards that have modern man coming on the scene 200,000 years ago.

In spite of all the evidence to the contrary, modern analysts confidently assert Reck’s Skeleton was of recent burial. It's rarely mentioned the records. The possibility of modern skeletons in such supposedly ancient strata is very destructive to modern scientific thought. Reck’s find is significant, it exposes the bias of the "science" of human origins, even 100 years past. Finds such as this undermine the long ages theory of Darwinian evolution, upon which modern anthropology, archeology and long-age geology origins theories are based.
Cremo, Michael A.; Thompson, Richard L. Forbidden Archaeology Torchlight Press 2011

New Mexico - Human Bones in Cavern
Bryan, William Alanson; Science, 70:39-41 1929
A cave was discovered by local treasure hunters on the lower slope of Bishops Cap by a Mr. Roscoe Conkling of El Paso in the 1920's. Intrigued by its soft floor, a party was formed and excavations began in the presence of a recognized authority able to witness any potential unearthing of items of importance. Digging down through the layers, animal bones now extinct began to be exposed and removed, then suddenly a human skull cap appeared at the 12 foot level. Further down the sediment layers a hard lens was encountered, and when broken through was found another skull and remains of numerous other animals including a camel at nearly 21 feet in depth. The scientist commented that this finding was of national significance, but for one reason or another, it never got into the textbooks.
Cremo, Michael A.; Thompson, Richard L. Forbidden Archaeology Torchlight Press 2011 p.444

For further studying:
- The Dorchester Pot, 534 million charlie-years old

- The Iron Bell, 300 million charlie-years old

- The South African Spheres, 2 800 million charlie-years old

- The Lanzhou Stone, 300 million charlie-years old

- Russian mechanical device found in volcanic rock, 400 million charlie-years old

- The Russian Ancient Screw, 300 million charlie-years old

- The ‘Guadeloupe Woman’, 28 million charlie-years old

- The London Hammer, 1 million charlie-years old

- The Zapata Track, 250 million charlie-years old

Cells compute ratios to control gene expression

Randomness has no role in biodiversity

https://phys.org/news/2017-05-cells-ratios-gene.html#nRlv

Excerpt: "In multicellular animals, cells communicate by emitting and receiving proteins, a process called signaling. One of the most common signaling pathways is the transforming growth factor b (Tgf-b) pathway, which functions in all animal species throughout their lifetimes and regulates numerous biological processes, such as instructing cells how to differentiate—whether a stem cell will become a muscle cell or a bone cell, for example.

But how do cells decipher those signals and use that information to guide gene expression? The answer, according to new research from the laboratory of Lea Goentoro, assistant professor of biology at Caltech: the cells perform simple division. In other words, they do math.

 

The cell's detection of the Tgf-b signal triggers a series of molecular interactions, culminating in changes in the abundance of a protein called Smad3. A kind of messenger, Smad3 is activated at the cell membrane when a cell encounters the Tgf-β signal, and ultimately ends up in the nucleus of the cell where it directs gene expression. Biologists have commonly believed that the degree to which gene expression is changed will depend on how much Smad protein is produced after exposure to Tgf-β.

In order to study how Smad responds to Tgf-b in real time, the researchers made movies of the signaling process in individual cells. They discovered that the pathway did not behave as biologists had previously thought. After exposure to the Tgf-b signal, the researchers found, Smad3 did indeed move to the nucleus, as expected. The surprise was that the abundance of Smad3 was significantly different in each cell. And yet, despite those wildly varying concentrations of Smad3, the level of Smad3 after the signal divided by the level before the signal was consistent in each cell.

This observation led the researchers to hypothesize that cells are somehow able to compute this ratio and that the gene response is proportional to this relative change (called a fold-change) rather than to the abundance of Smad3.

"This result is nonintuitive: it is easy to imagine how a cell can increase the expression of a target gene by increasing the abundance of Smad3," says Frick. "Thus, many of us had expected that the abundance of Smad3 would be tuned in response to the Tgf-β signal. In contrast, what these cells regulate is the ratio of the change in abundance of Smad3. The big question now is: How does a cell compute this ratio and then adjust gene expression accordingly?"

My comment: Cellular mechanisms are extremely complex. Signaling pathways, readers, writers and erasers point to design. Several forms of information (Digital, analog and meta-data) in the cell tell us about living machines that are much more complex than human designed computers. Randomness has no role in biodiversity. 

Why scientists are in a hurry to develop genetic repairing methods

If all of the people with rare genetic diseases lived in one country, it would be the world’s 3rd most populous country

https://globalgenes.org/rare-diseases-facts-statistics/

Statistics and Figures on Prevalence of Genetic and Rare Diseases

Although rare and genetic diseases, and many times the symptoms, are uncommon to most doctors, rare diseases as a whole represent a large medical challenge. Combine this with the lack of financial or market incentives to treat or cure rare diseases, and you have a serious public health problem.
Here are a few statistics and facts to illustrate the breadth of the rare disease problem worldwide:

- There are approximately 7,000 different types of rare diseases and disorders, with more being discovered each day
- 30 million people in the United States are living with rare diseases. This equates to 1 in 10 Americans or 10% of the U.S. population
- Similar to the United States, Europe has approximately 30 million people living with rare diseases. It is estimated that 350 million people worldwide suffer from rare diseases
- If all of the people with rare diseases lived in one country, it would be the world’s 3rd most populous country
- In the United States, a condition is considered “rare” it affects fewer than 200,000 persons combined in a particular rare disease group. International definitions on rare diseases vary. For example in the UK, a disease is considered rare if it affects fewer than 50,000 citizens per disease
- 80% of rare diseases are genetic in origin, and thus are present throughout a person’s life, even if symptoms do not immediately appear
- Approximately 50% of the people affected by rare diseases are children
- 30% of children with rare disease will not live to see their 5th birthday
- Rare diseases are responsible for 35% of deaths in the first year of life
- The prevalence distribution of rare diseases is skewed – 80% of all rare disease patients are affected by approximately 350 rare diseases
- According to the Kakkis EveryLife Foundation, 95% of rare diseases have not one single FDA approved drug treatment
- During the first 25 years of the Orphan Drug Act (passed in 1983), only 326 new drugs were approved by the FDA and brought to market for all rare disease patients combined
- According to the National Institutes of Health Office of Rare Disease Research, approximately 6% of the inquiries made to the Genetic and Rare Disease Information Center (GARD) are in reference to an undiagnosed disease
- Approximately 50% of rare diseases do not have a disease specific foundation supporting or researching their rare disease

My comment: If you see someone claiming about random mutations and evolution, please correct the claims politely and ask him/her to stop pseudoscience. There is no such a thing as a mutation driven evolution or natural selection. 

There are no mechanisms for macroevolution

Epigenetic markers strongly affect binding of transcription factors

http://www.genengnews.com/gen-news-highlights/epigenetic-marks-shun-some-transcription-factors-embrace-others/81254309?utm_medium=newsletter

Excerpt: "The same epigenetic marks can be read as “keep off” or “welcome,” depending on what DNA-binding protein, or transcription factor, is doing the reading. These marks, methylated cytosine and guanine dinucleotides (mCpGs), normally indicate which portions of the genome are inactive. But new findings from a systematic study of hundreds of transcription factors suggest that mCpGs may play a more subtle role in gene regulation.

In this new study, scientists based at Karolinska Institutet systematically analyzed the binding specificities of transcription factors to DNA that was marked by mCpGs, as well as to DNA that was unmarked by mCpGs. The observed that mCpGs can influence binding of most transcription factors to DNA—in some cases negatively and in others positively.

Interestingly, many of the transcription factors that prefer to bind to mCpG sites appear to be important to development. This finding may inform future analyses of the role of DNA methylation on cell differentiation, chromatin reprogramming, and transcriptional regulation.

The results pave the way for cracking the genetic code that controls the expression of genes, and will have broad implications for the understanding of development and disease. The availability of genomic information relevant to disease is expanding at an exponentially increasing rate.

"This study identifies how the modification of the DNA structure affects the binding of transcription factors, and this increases our understanding of how genes are regulated in cells and further aids us in deciphering the grammar written into DNA," noted Professor Taipale.""

Excerpt from the original study:

"One shortcoming is the lack of knowledge about DNA binding specificities (motifs) for hundreds of the estimated ~1600 human transcription factors. Another is how transcription factor binding is modulated by “epigenetics”—a contentious term that refers to heritable states of both cells and organisms, as well as the covalent chemical modifications of DNA and protein that often provide the underlying mechanism. DNA methylation at cytosine and guanine dinucleotides (mCG) satisfies most views of epigenetics, as it is inherited across cell divisions and functions in imprinting (parent-of-origin–dependent gene expression). On page 502 of this issue, Yin et al. provide a comprehensive look at the extent to which human transcription factor binding is affected by mCG, and make a striking finding: Many homeodomain transcription factors—perhaps the best characterized developmental regulators in biology can bind to specific methylated DNA sequences.

More generally, the study of Yin et al. contributes a unique perspective to evaluating how transcription factors bind DNA. Transcription factor motif modeling is critical for the study of global gene regulation, allowing us to predict potential binding sites in the genome. Given that so many transcription factors are affected by chemical modifications to DNA, we are now faced with a clear necessity to incorporate DNA methylation into motif models, and a new type of data from which to learn to read the genome the way transcription factors do."

My comment: This study confirms my previous claims about factors affecting phenotypes of organisms and observed changes in nature. Alterations in organisms are based on epigenetic factors and designed mechanisms mediated by nutrition, stress, climate and other environmental factors. Even one addition or deletion of one methyl group on a gene might have a significant influence on cell activity and identity. Random mutations or natural selection have no role in biodiversity. There are no mechanisms for large scale evolution. Everything points to God's design and creation.

P.S. Thanks for Jorge Rodriguez for sending me the original paper.

Forget about designer babies - You can screen for genetic diseases but not for traits

Forget about designer babies - You can screen for genetic diseases but not for traits

https://phys.org/news/2017-02-ethicists-day-heredity-disease.html

Excerpt: "Don't expect designer babies any time soon—but a major new ethics report leaves open the possibility of one day altering human heredity to fight genetic diseases, with stringent oversight, using new tools that precisely edit genes inside living cells.

What's called genome editing already is transforming biological research, and being used to develop treatments for patients struggling with a range of diseases.

The science is nowhere near ready for a huge next step that raises ethical questions—altering sperm, eggs or embryos so that babies don't inherit a disease that runs in the family, says a report Tuesday from the National Academy of Sciences and National Academy of Medicine.

But if scientists learn how to safely pass alterations of the genetic code to future generations, the panel said "germline" editing could be attempted under strict criteria, including that it targets a serious disease with no reasonable alternative and is conducted under rigorous oversight."

https://www.geneticliteracyproject.org/2017/05/03/mendel-meets-tinder-genepeeks-previde-screen-maps-babies-genetic-risks-conception/#.WQpOZYfLz_B.twitter

Excerpt: "Another limitation of gene variant databases such as ClinVar is the confusing “variants of uncertain significance” (VUS) that tell a patient little more than that yes, your gene variant isn’t the “normal” one, but no, it hasn’t been associated with symptoms. Yet. VUS are often of unknown impact because not enough cases have been reported to statistically establish a mutation-disease connection. And that’s where GenePeeks’s deeper dive comes in. “We don’t wait for a variant to show up in a patient, but validate our analysis on a gene-by-gene basis” considering the effect on the protein, Silver said."

My comment: Gene sequences don't determine heritable traits. That's why by modifying the gene sequences only, scientists can't affect human characteristics. Every cell inside you is having the exact same gene sequences. For successful cellular differentiation and for building thousands of different cell types there are other forms of biological information in the cell. This type of information is called epigenetic control of gene expression. Genetic mutations never lead to evolution or adaptation. They cause genetic degradation, disorders and diseases. Modern science is aware of 200,000 disease causing genetic mutations in the human DNA. 10,000 new genetic defects are discovered every year.

http://www.nature.com/news/past-5-000-years-prolific-for-changes-to-human-genome-1.11912#/b3

Excerpt: "Of 1.15 million single-nucleotide variants found among more than 15,000 protein-encoding genes, 73% in arose the past 5,000 years, the researchers report."

My comment: All data points to biblical creation. Don't get lost.