Gene Therapy Advance

An important methodological advance that can be of use for gene therapy is described here.  This is a modification of the CRISPR system that now allows targeted base editing of DNA with high efficiency and low off-target effects.  In practical terms, this would allow targeted correction of DNA mutations, assuming an efficient and safe delivery system could be devised.  These are exciting days for the development of potential gene therapy approaches.  Abstract:

The spontaneous deamination of cytosine is a major source of C•G to T•A transitions, which account for half of known human pathogenic point mutations. The ability to efficiently convert target A•T base pairs to G•C could therefore advance the study and treatment of genetic diseases. While the deamination of adenine yields inosine, which is treated as guanine by polymerases, no enzymes are known to deaminate adenine in DNA. Here we report adenine base editors (ABEs) that mediate conversion of A•T to G•C in genomic DNA. We evolved a tRNA adenosine deaminase to operate on DNA when fused to a catalytically impaired CRISPR-Cas9. Extensive directed evolution and protein engineering resulted in seventh-generation ABEs (e.g., ABE7.10), that convert target A•T to G•C base pairs efficiently (~50% in human cells) with very high product purity (typically ≥ 99.9%) and very low rates of indels (typically ≤ 0.1%). ABEs introduce point mutations more efficiently and cleanly than a current Cas9 nuclease-based method, induce less off-target genome modification than Cas9, and can install disease-correcting or disease-suppressing mutations in human cells. Together with our previous base editors, ABEs advance genome editing by enabling the direct, programmable introduction of all four transition mutations without double-stranded DNA cleavage.

"Mutation-Independent" Cancer Therapy: Targeting Mitochondria

By Kelvinsong - Own work, CC0, https://commons.wikimedia.org/w/index.php?curid=27715320

Mitochondria are the “powerhouses of the cell” and problems with mitochondria have been associated with cancer.  So, putting stress on mitochondria can mimic the effects of aberrant gene expression to stimulate breast cancer stem cell activity.  This effect can be inhibited by the antibiotic doxycycline, demonstrating that “mutation-independent” cancer therapy approaches can be used against cancer – approaches that target such phenomena such as mitochondrial function and reproduction, or some other aspects of cell phenotype, as opposed to targeting specific gene mutations and the aberrant products of such mutated genes.  Abstract:

Here, we used MCF7 cells as a model system to interrogate how MYC/RAS co-operativity contributes to metabolic flux and stemness in breast cancer cells. We compared the behavior of isogenic MCF7 cell lines transduced with c-Myc or H-Ras (G12V), either individually or in combination. Cancer stem cell (CSC) activity was measured using the mammosphere assay. c-Myc augmented both mammosphere formation and mitochondrial respiration, without any effects on glycolytic flux. In contrast, H-Ras (G12V) synergistically augmented both mammosphere formation and glycolysis, but only in combination with c-Myc, directly demonstrating MYC/RAS co-operativity. As c-Myc is known to exert its effects, in part, by stimulating mitochondrial biogenesis, we next examined the effects of another stimulus known to affect mitochondrial biogenesis, i.e. ROS production. To pharmacologically induce oxidative stress, we used Rotenone (a mitochondrial inhibitor) to target mitochondrial complex I. Treatment with Rotenone showed bi-phasic effects; low-dose Rotenone (1 to 2.5 nM) elevated mammosphere formation, while higher doses (10 to 100 nM) were inhibitory. Importantly, the stimulatory effects of Rotenone on CSC propagation were blocked using a mitochondrial-specific anti-oxidant, namely Mito-tempo. Thus, "mild" mitochondrial oxidative stress, originating at Complex I, was sufficient to pheno-copy the effects of c-Myc, effectively promoting CSC propagation. To validate the idea that mitochondrial biogenesis is required to stimulate CSC propagation, we employed Doxycycline, a well-established inhibitor of mitochondrial protein translation. Treatment with Doxycycline was indeed sufficient to block the stimulatory effects of H-Ras (G12V), c-Myc, and Rotenone on CSC propagation. As such, Doxycycline provides a strong rationale for designing new therapeutics to target mitochondrial biogenesis, suggesting a new "mutation-independent" approach to cancer therapy. In support of this notion, most currently successful anti-cancer agents therapeutically target "cell phenotypes", such as increased cell proliferation, rather than specific genetic mutations. Remarkably, we demonstrated that Doxycycline inhibits the effects of diverse oncogenic stimuli, of both i) genetic (MYC/RAS) and ii) environmental (Rotenone) origins. Finally, we discuss the advantages of our "Proteomics-to-Genomics (PTG)" approach for in silico validation of new biomarkers and novel drug targets. In this context, we developed a new Myc-based Mito-Signature consisting of 3 mitochondrial genes (HSPD1; COX5B; TIMM44) for effectively predicting tumor recurrence (HR=4.69; p=2.4e-08) and distant metastasis (HR=4.94; p=2.8e-07), in ER(+) in breast cancer patients. This gene signature could serve as a new companion diagnostic for the early prediction of treatment failure in patients receiving hormonal therapy.

The FDA And Vasopressin

The FDA got involved in trying to regulate marketed previously unapproved drugs, with results perhaps not unexpected to those of us who have taken a MBA-level course in economics.  From the (publicly available) paper linked above:

In May 2017, the US Food and Drug Administration (FDA) announced a Drug Competition Action Plan, designed to address competition and pricing in the generic market and improve access to prescription drugs. One of FDA’s stated goals is to reexamine “places where its rules—including standards and procedures related to generic drug approvals—are being used in ways that may create obstacles to generic access,” instead of ensuring the vigorous competition Congress intended. In this Viewpoint, we examine FDA’s 2006 Unapproved Drugs Initiative (UDI), designed to strengthen the agency’s regulatory oversight related to unapproved marketed drugs. Using an illustrative example, we discuss this initiative’s unintended consequences, as it appears to have created obstacles to generic drug access, likely increasing prescription drug costs…On November 14, 2014, Par received FDA approval for Vasostrict, and on December 15, 2014, FDA instructed all other suppliers of unapproved intravenous vasopressin to stop manufacturing their products by January 30, 2015, leaving only Par with a marketed product. Subsequently, the average wholesale price of intravenous vasopressin increased from $4.27 to $138.40 per vial in November 2016, a 3141% increase. In 2013, when there were multiple competing suppliers, total sales from intravenous vasopressin approximated $4 million. As of November 2016, Vasostrict achieved annualized sales of nearly $400 million. As a result of the high cost, reports have surfaced of vasopressin being removed from code carts, making it unavailable in life-threatening situations.

I’ll leave it to the reader to ponder whether the vast increase per vial price for vasopressin was a justifiable change, and whether the unavailability of this drug for endangered patients is somehow compatible with the FDA’s ostensible mission.

The FDA and Big Pharm: a match made in heaven…or hell.  You decide.  Look at the facts reported in the paper and the other link and ponder well.

Gut Microbiota And Metastasis

Remarkably, not only are primary colorectal cancers colonized by particular strains of bacteria from the gut microbiota, but distal metastases of those tumors have the same strains as well.  Therefore, tumor-associated bacteria “go along for the ride” when metastasis occurs; in particular, Fusobacterium nucleatum is associated with both the primary and metastatic tumors, illustrating stability of bacteria-cancer association.  Mouse xenografts of human primary colorectal cancers exhibited maintenance of the bacteria, and antibiotic treatment “reduced Fusobacterium load, cancer cell proliferation and overall tumor growth.”  One wonders if the bacterial actually promote metastasis as well, and are not just “along for the ride;” it would seem reasonable to suppose that at least the bacteria help the metastasis get established and grow at its new location.  All of this highlights the link between gut microbiota and cancer; note also that diet affects the microbiota, possibly in a clinically-relevant manner.  Abstract:

Colorectal cancers comprise a complex mixture of malignant cells, non-transformed cells and microorganisms. Fusobacterium nucleatum is among the most prevalent bacterial species in colorectal cancer tissues. Here we show that colonization of human colorectal cancers with Fusobacterium and its associated microbiome, including Bacteroides, Selenomonas and Prevotella species, is maintained in distal metastases, demonstrating microbiome stability between paired primary-metastatic tumors. In situ hybridization analysis revealed that Fusobacterium is predominantly associated with cancer cells in the metastatic lesions. Mouse xenografts of human primary colorectal adenocarcinomas were found to retain viable Fusobacterium and its associated microbiome through successive passages. Treatment of mice bearing a colon cancer xenograft with the antibiotic metronidazole reduced Fusobacterium load, cancer cell proliferation and overall tumor growth. These observations argue for further investigation of antimicrobial interventions as a potential treatment for patients with Fusobacterium-associated colorectal cancer.

Tired of mainstream media

I have re-started my "inside walking" in the evenings. 

In good weather, I always walk outside after dinner. However, once the weather deteriorates, and we switch to winter time, it is already dark and sometimes, extremely cold.

So, after a full workday and dinner, I have no other choice but to walk the periphery of my combined kitchen-living room space.  I walk minimum for 35 minutes every evening, and at the same time, I listen to podcasts. 

I have not turned on the TV for ages. I do not remember the last time I watched the news. If I happen to be close to a TV outside my home, I feel that its constant chatter tires me, bores me and even irritates me.

These days, I thought that I have already heard enough from the financial independence guys, so I started looking for other podcasts that would keep me entertained while walking. 

I was lucky to find something interesting. The podcasts are by men, and yet, the topics are of interest to me. The speakers are honest, which is refreshing, and all of them have great experiences to relate.

The Jocko Podcast is my first choice. Below is one great Jocko episode that I listened to just today.

Another recommendation is the podcast of Quintus Curtius. One of the episodes that hit close to home was this one:

Of course, I still follow up on any recent podcast by Joshua Sheats. He has been "my walking buddy" for more than a year now, and he still inspires me to go on.

Well, tomorrow is work as usual. I had four days of rest, and I feel slightly recharged.

Good luck to all working people out there, and I hope that you can find inspiration and thought-provoking topics on the podcasts of Jocko, Quintus Curtius and Joshua Sheats!

Why is sugar bad for you

I just found this great webpage with simple explanations of why sugar (50% glucose, 50% fructose) is bad for you. The information is unfortunately unknown to most practicing clinicians and to our medical students. This is why medical students and faculty still feast on this:
 





Educate yourself, do not rely on your doctor!

Deck the Hallways

I decided to resort to my backyard for Christmas 2017 decorations. Yesterday, the weather cooperated with my intention, and I foraged for everything that was still green. The result is below. 

More On Cancer Prevention And Lifestyle

Here is recent news on yet another study linking lifestyle to cancer risk.  These are all things you, as a reader of this blog, should be well aware of by now.  The importance of curbing these behaviors to reduce cancer risk is underscored by these additional data.

Researchers with the American Cancer Society looked at data on cancer incidence and deaths, finding that 42 percent of all cancer cases in the United States -– and nearly half of all cancer deaths – are linked to preventable risk factors like cigarette smoking, exposure to secondhand smoke, excess body weight, alcohol intake and dietary choices.

Cigarette smoking, in particular, was connected to far more cancer cases and deaths than any other single risk factor, accounting for 19 percent of all cancer cases and 28.8 percent of deaths. Overweight and obesity came in second, responsible for 7.8 percent of cases and 6.5 percent of deaths, while alcohol intake was the third most important factor, leading to 5.6 percent of cancer cases and 4 percent of deaths.

Although there are some sex-specific differences as to the relative importance of various risk factors, the bottom line is that these things – smoking/overweight/obese, bad diets, certain STDs, alcohol – are bad for everyone.  Be aware and adjust your life accordingly, to the extent you are able.

Four Approaches To Weight Loss

By FatM1ke - Central_Obesity_011.jpgCentral_Obesity_008.jpg, Public Domain, https://commons.wikimedia.org/w/index.php?curid=4412552

Here is a review paper on weight loss that focuses on four methods: (1) juicing/detoxification, (2) intermittent fasting, (3) paleo, and (4) high intensity training.  The first three methods all led to weight loss due to caloric restriction; however, the extreme “fad diets” of juicing or detoxification lead to weight gain when the normal diet is resumed. Paleo, if followed continuously, would seem to enable the weight to be kept off; the basic lesson is that a diet needs to be a sustainable, long-term commitment, not a short-term extreme such as, for example, juicing. Intermittent fasting also can work if followed on a regular basis.  I myself cut back on saturated fat, “junk” food and excess sugar, while increasing fruit, whole grains, and vegetables (with one “cheat day” in which I really don’t eat that unhealthy either).  Caloric restriction is what all have in common.  Interestingly, “high intensity training” worked as well, despite the recent paradigm that says that weight loss is overwhelmingly about diet and not exercise (the latter of which has a host of other benefits). Perhaps the type of exercise is important, and one wonders if all variables are taken into account; perhaps those who do “high intensity training” have healthier diets.  Abstract:

PURPOSE OF REVIEW:

The purpose of this paper is to review the epidemiology of obesity and the most recent literature on popular fad diets and exercise regimens that are used for weight loss. The weight loss plans that will be discussed in this article include juicing or detoxification diets, intermittent fasting, the paleo diet, and high intensity training.

RECENT FINDINGS:

Despite the growing popularity of fad diets and exercise plans for weight loss, there are limited studies that actually suggest these particular regimens are beneficial and lead to long-term weight loss. Juicing or detoxification diets tend to work because they lead to extremely low caloric intake for short periods of time, however tend to lead to weight gain once a normal diet is resumed. Both intermittent fasting and the paleo diet lead to weight loss because of overall decreased caloric intake as well. Lastly, studies on short bursts of high intensity training have shown remarkable weight loss and improvements in cardiovascular health. Review of the literature does suggest that some fad diets and exercise plans do lead to weight loss; however, the studies are quite limited and are all based on the concept of caloric restriction.

BUTTERNUT SQUASH DESSERT

It is creamy, sweet and unbelievably healthy!
The recipe came to be because I had some baked butternut squash frozen from a month ago, and it was Thanksgiving. Here it is:

Ingredients:
one butternut squash, cut in half and baked
one banana
one tsp vanilla
a dash of cinnamon
a few Tbsp almond milk

Instructions:
Peel the baked butternut squash and pulse in a blender together with the banana and some almond milk to the desired consistency. Mix in the vanilla and cinnamon, chill. Serve with toasted nuts on top.

Happy Thanksgiving 2017

By Jean Bungartz - This file has been extracted from another file: Hühner (Geflügel-Album, Jean Bungartz, 1885).pdfKarlsruhe: Badische Landesbibliothek, 2013http://digital.blb-karlsruhe.de/urn/urn:nbn:de:bsz:31-38086, Public Domain, https://commons.wikimedia.org/w/index.php?curid=27040412 

Not for the Thanksgiving meal table?

Who will eat who?  Just joking…but those are huge chickens. Happy Thanksgiving.

Cancer-Immune Set Point

Why do patients have variable responses to cancer immunotherapy?  Abstract:

Immunotherapy is proving to be an effective therapeutic approach in a variety of cancers. But despite the clinical success of antibodies against the immune regulators CTLA4 and PD-L1/PD-1, only a subset of people exhibit durable responses, suggesting that a broader view of cancer immunity is required. Immunity is influenced by a complex set of tumour, host and environmental factors that govern the strength and timing of the anticancer response. Clinical studies are beginning to define these factors as immune profiles that can predict responses to immunotherapy. In the context of the cancer-immunity cycle, such factors combine to represent the inherent immunological status - or 'cancer-immune set point' - of an individual.

It is always important to consider individual variation when judging therapeutic effectiveness or disease risk.  Different people have varying "immune profiles" - what the authors term the "cancer-immune set point" - and these different profiles will influence how each patient reacts to cancer immunotherapy.  Indeed, it is useful to speculate that these different set points affect how the patient's immune system reacts to a developing cancer in the first place - why in some individuals the early-stage cancer cells will be eliminated by the immune system and in other individuals the cancer cells are not recognized and grow to a full-fledged disease state.  Similarly, when you hear that "my grandfather smoked for 80 years and never got cancer and lived to be 96" or that "my grandma ate bacon-wrapped cheeseburgers three times a day for 70 years and died in her sleep at age 88" remember that there is individual variation and that we talk about disease risk - probabilities - not that lifestyle impacts disease to have a 100% deterministic effect on every person.  In like manner, immunotherapy may be generally effective, but it can not be expected to be equally effective for every individual.  Possibly, by understanding the mechanisms behind these differences, the effectiveness of immunotherapy can be enhanced even for relatively resistant individuals.

Thank yourself first and find your mission

Yesterday, I was listening to an interview with Tim Ferriss. He mentioned his idea that by inviting voluntary suffering to our lives, we are preparing ourselves for the times when we face involuntary suffering. 

I accept the concept, but I cannot practice it since my current life is already full of involuntary unpleasantness (not exactly suffering, I admit). However, when I have to deal with unpleasantness every second of my wakeful hours and even my dreams are molded by the daily horrors, I have no time to invite suffering on top of everything. When you put out fires all day, you do not start one. 

Tim’s advice is good for people of leisure who have time and money and seek some antidote to their cushy life. Instead of drugs, bring some edge to your life by experiencing deprivation, cold, and maybe hunger! I approve.

The other notable moment in the interview was the thought that most of us are reactive, rather than proactive in everyday life. As a result, creativity and building up our individuality are crushed. How many of us have time to embark on self-assigned and self-initiated projects every day or even at all? This reactive way of existence has been another problem of mine for the past months. I have not had time to do something that I have designed, thought about, craved, envisioned. I have been losing sight of my identity. 

I need to go back to some foundational work in my mind and decide on what my mission is. Yes, everyone of us should decide on what their mission is. The mission can be re-visited, re-evaluated and re-adjusted with time, but we need to remember what we are doing with our precious little time on this earth. 

On this Thanksgiving, I am thankful that I exist!  I am also grateful for having time to muse about the meaning of my existence. Is this too egotistical? Not at all. If you appreciate yourself and find your true identity, you would be able to help others in a meaningful way. If you are miserable, feel bad about yourself, the only thing you can bring in people's life is misery and disaster. So, celebrate yourself, your freedom and the precious days when you have the time to realize what you stand for.  Today, I will cook for many hours, but this will be a proactive expression of myself. As far as my work unpleasantness goes, I should remember this quote from this link: “Those that matter don’t mind, and those that mind don’t matter.” 


Recently, I had also found some solace in this great talk by Martin Rossman. 

Happy Thanksgiving to all!

Wnt Signaling And Multiple Sclerosis

By Quasar Jarosz at English Wikipedia, CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=7616130

The paper linked here asserts that downregulation of Wnt signaling (also involved in many types of cancer) is essential for proper neuronal differentiation, and that activated Wnt signaling can impair remyelination of neurons. Since demyelination – loss of the myelin sheath of neurons that is important for nerve signal conduction – leads to multiple sclerosis, this finding points the way for possible therapeutic interventions against that disease.  Abstract:

Multiple sclerosis (MS) is marked by neuroinflammation and demyelination with loss of oligodendrocytes in the central nervous system. The immune response is regulated by WNT/beta-catenin pathway in MS. Activated NF-kappaB, a major effector of neuroinflammation, and upregulated canonical WNT/beta-catenin pathway positively regulate each other. Demyelinating events present an upregulation of WNT/beta-catenin pathway, whereas proper myelinating phases show a downregulation of WNT/beta-catenin pathway essential for the promotion of oligodendrocytes precursors cells proliferation and differentiation. The activation of WNT/beta-catenin pathway results in differentiation failure and impairment in remyelination. However, PI3K/Akt pathway and TCF7L2, two downstream targets of WNT/beta-catenin pathway, are upregulated and promote proper remyelination. The interactions of these signaling pathways remain unclear. PPAR gamma activation can inhibit NF-kappaB, and can also downregulate the WNT/beta-catenin pathway. PPAR gamma and canonical WNT/beta-catenin pathway act in an opposite manner. PPAR gamma agonists appear as a promising treatment for the inhibition of demyelination and the promotion of proper remyelination through the control of both NF-kappaB activity and canonical WNT/beta-catenin pathway.

Taste Bud Wiring

Here is an interesting basic science paper about how the taste bud sensors are “wired” to detect the basic taste sensations.  Abstract:

In mammals, taste buds typically contain 50-100 tightly packed taste-receptor cells (TRCs), representing all five basic qualities: sweet, sour, bitter, salty and umami. Notably, mature taste cells have life spans of only 5-20 days and, consequently, are constantly replenished by differentiation of taste stem cells. Given the importance of establishing and maintaining appropriate connectivity between TRCs and their partner ganglion neurons (that is, ensuring that a labelled line from sweet TRCs connects to sweet neurons, bitter TRCs to bitter neurons, sour to sour, and so on), we examined how new connections are specified to retain fidelity of signal transmission. Here we show that bitter and sweet TRCs provide instructive signals to bitter and sweet target neurons via different guidance molecules (SEMA3A and SEMA7A). We demonstrate that targeted expression of SEMA3A or SEMA7A in different classes of TRCs produces peripheral taste systems with miswired sweet or bitter cells. Indeed, we engineered mice with bitter neurons that now responded to sweet tastants, sweet neurons that responded to bitter or sweet neurons responding to sour stimuli. Together, these results uncover the basic logic of the wiring of the taste system at the periphery, and illustrate how a labelled-line sensory circuit preserves signalling integrity despite rapid and stochastic turnover of receptor cells

Plant Extract Against Colon Cancer?

Anti-cancer benefits of a plant extract, mediated through inhibition of Wnt signaling. This may be useful for developing preventive or therapeutic approaches to colon cancer.  Abstract:

OBJECTIVE:

To investigate the effect of the ethanol extract of Scutellaria barbata D. Don (EESB) on colorectal cancer (CRC) growth and Wnt/β-catenin signaling pathway in vivo and in vitro.

METHODS:

In vivo experiment, CRC xenograft mouse model was constructed with injection of HT-29 cells. Following xenograft implantation, twenty mice were randomly divided into EESB-treated group (n=10) and control group (n=10) by a random number table, and were given with intra-gastric administration of 2 g/kg EESB or saline, 5 days a week for 16 days, respectively. At the end of experiment, tumors were removed and weighed by electronic scales. The proliferation biomarker Ki-67 of tumor was evaluated by immunohistochemistry (IHC) assay. In vitro study, HT-29 cells were treated with 0, 0.5, 1.5, 2.5 mg/mL EESB for 24 h. At the end of the treatment, the viability and survival of HT-29 cells were determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and colony formation assay, respectively. The mRNA expression of c-Myc, Survivin and adenomatous polyposis coli (APC) was examined by reverse transcription-polymerase chain reaction (RT-PCR) both in tumor tissues of CRC xenograft mice and HT-29 cells. Protein expression of c-Myc, Survivin, APC, and β-catenin as well as β-catenin phosphorylation level were evaluated by IHC assay or Western blotting.

RESULTS:

EESB significantly reduced tumor weight in CRC xenografts mice, compared with the control group (P<0.05). IHC assay showed that EESB significantly inhibited protein expression of Ki-67 in tumor tissues (P<0.05). MTT assay showed that EESB significantly reduced HT-29 cell viability in a dose-dependent manner (P<0.05). Colony formation assay showed that EESB dose-dependently decreased the survival of HT-29 cells (P<0.05). In addition, RT-PCR assay showed that EESB decreased the mRNA expression of c-Myc and Survivin and increased APC expression, both in tumor tissues of CRC xenograft mice and HT-29 cells (P<0.05). IHC assay or Western blotting showed that EESB decreased protein expression of β-catenin, c-Myc and Survivin, as well as increased APC expression and β-catenin phosphorylation in tumor tissues or HT-29 cells (P<0.05).

CONCLUSIONS:

EESB significantly reduced tumor growth in CRC xenografts mice, and inhibited the viability and survival of HT-29 cells. EESB could suppress the activation of the Wnt/β-catenin pathway, which might be one of the mechanisms whereby Scutellaria barbata D. Don exerts its anticancer activity.

Inflammation Memory

By The original uploader was Samir at English Wikipedia - Transferred from en.wikipedia to Commons., CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=2107303

Here is an interesting study that suggests a form of “inflammation memory” exists.  Therefore, in epithelial stem cells that have already been exposed to acute inflammation there is a molecular “memory” of the event, which enables a more rapid response for subsequent inflammatory exposures.   While this has some benefits in dealing with the immediate need for a response to certain stimuli, this increased sensitivity may contribute to autoimmune diseases and cancer.  From the preventive and therapeutic angle, it is therefore important to study.  Abstract:

The skin barrier is the body's first line of defence against environmental assaults, and is maintained by epithelial stem cells (EpSCs). Despite the vulnerability of EpSCs to inflammatory pressures, neither the primary response to inflammation nor its enduring consequences are well understood. Here we report a prolonged memory to acute inflammation that enables mouse EpSCs to hasten barrier restoration after subsequent tissue damage. This functional adaptation does not require skin-resident macrophages or T cells. Instead, EpSCs maintain chromosomal accessibility at key stress response genes that are activated by the primary stimulus. Upon a secondary challenge, genes governed by these domains are transcribed rapidly. Fuelling this memory is Aim2, which encodes an activator of the inflammasome. The absence of AIM2 or its downstream effectors, caspase-1 and interleukin-1β, erases the ability of EpSCs to recollect inflammation. Although EpSCs benefit from inflammatory tuning by heightening their responsiveness to subsequent stressors, this enhanced sensitivity probably increases their susceptibility to autoimmune and hyperproliferative disorders, including cancer.

Do you need to let go

Today I am sick, sitting in a chair next to the window and looking at the remaining tree leaves. They are all golden-brown and moving with the wind. Soon there will be only naked branches to look at.

I am also listening to Enya:

Enya's music is like a stream that purifies, flows through you and allows you to let go, let go of worries, pain, and pettiness.

Wool Underwear For Lower Back Pain

Public Domain, https://commons.wikimedia.org/w/index.php?curid=16818

Lower back pain is a common complaint; here we see a study suggesting that the use of wool underwear leads to "significant improvements in pain intensity, disability, and lower back flexibility." Temperature-related mechanism? Abstract:

The aim was to assess the effect of wool underwear use in patients with chronic non specific low back pain. The study employed two-group, experimental design. A total of 48 patients with chronic non specific low back pain were selected for the study. They were distributed into two groups: a control group and a treatment group. The 24 patients in each group were randomly selected and the compositions of the two groups. The patients in the treatment group wore woolen underwear during the experimental period of 2 month. All patients were assessed at the beginning the trial (pre-test) and the end of 8th (post-test) week. Data were collected using the visual analogue pain scale, Oswestry Disability Index and Schober test measurements. Patients in the treatment group reported significant improvements in their conditions including a reduction in pain levels and Oswestry Disability Index, and Schober test measurements increased (p<0.001). Patients with chronic non-specific low back pain who wore wool underwear experienced significant improvements in pain intensity, disability, and lower back flexibility.

Caveat emptor.  

Potential Stem Cell Therapeutic Engineering

This paper discusses some advances in genetic engineering of cells, which may be of use “to develop innovative stem cell-based therapeutics.”  The link between basic science and therapeutic advances remains strong.  Abstract:

Precise and efficient manipulation of genes is crucial for understanding the molecular mechanisms that govern human hematopoiesis and for developing novel therapies for diseases of the blood and immune system. Current methods do not enable precise engineering of complex genotypes that can be easily tracked in a mixed population of cells. We describe a method to multiplex homologous recombination (HR) in human hematopoietic stem and progenitor cells and primary human T cells by combining rAAV6 donor delivery and the CRISPR/Cas9 system delivered as ribonucleoproteins (RNPs). In addition, the use of reporter genes allows FACS-purification and tracking of cells that have had multiple alleles or loci modified by HR. We believe this method will enable broad applications not only to the study of human hematopoietic gene function and networks, but also to perform sophisticated synthetic biology to develop innovative engineered stem cell-based therapeutics.

Spinal Muscular Atrophy Gene Therapy Clinical Trial

By Kashmiri, based on earlier work by Domaina - Own work based on Autosomal dominant - en.svg and Autorecessive.jpg, CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=19018894

Spinal muscular atrophy is an extremely serious neurological disease of childhood that is inherited in an autosomal recessive manner.  There has recently been an encouraging gene therapy clinical trial for this disease.  Families of children with this disorder need to be made aware of these findings.  Note: "...a single intravenous infusion of adeno-associated viral vector containing DNA coding for SMN resulted in longer survival, superior achievement of motor milestones, and better motor function than in historical cohorts."  Abstract:

BACKGROUND:

Spinal muscular atrophy type 1 (SMA1) is a progressive, monogenic motor neuron disease with an onset during infancy that results in failure to achieve motor milestones and in death or the need for mechanical ventilation by 2 years of age. We studied functional replacement of the mutated gene encoding survival motor neuron 1 (SMN1) in this disease.

METHODS:

Fifteen patients with SMA1 received a single dose of intravenous adeno-associated virus serotype 9 carrying SMN complementary DNA encoding the missing SMN protein. Three of the patients received a low dose (6.7×1013 vg per kilogram of body weight), and 12 received a high dose (2.0×1014 vg per kilogram). The primary outcome was safety. The secondary outcome was the time until death or the need for permanent ventilatory assistance. In exploratory analyses, we compared scores on the CHOP INTEND (Children's Hospital of Philadelphia Infant Test of Neuromuscular Disorders) scale of motor function (ranging from 0 to 64, with higher scores indicating better function) in the two cohorts and motor milestones in the high-dose cohort with scores in studies of the natural history of the disease (historical cohorts).

RESULTS:

As of the data cutoff on August 7, 2017, all 15 patients were alive and event-free at 20 months of age, as compared with a rate of survival of 8% in a historical cohort. In the high-dose cohort, a rapid increase from baseline in the score on the CHOP INTEND scale followed gene delivery, with an increase of 9.8 points at 1 month and 15.4 points at 3 months, as compared with a decline in this score in a historical cohort. Of the 12 patients who had received the high dose, 11 sat unassisted, 9 rolled over, 11 fed orally and could speak, and 2 walked independently. Elevated serum aminotransferase levels occurred in 4 patients and were attenuated by prednisolone.

CONCLUSIONS:

In patients with SMA1, a single intravenous infusion of adeno-associated viral vector containing DNA coding for SMN resulted in longer survival, superior achievement of motor milestones, and better motor function than in historical cohorts. Further studies are necessary to confirm the safety and efficacy of this gene therapy. (Funded by AveXis and others; ClinicalTrials.gov number, NCT02122952 .).