**Biohacking: Methods, Practices, and Supporting Evidence**

**What is Biohacking?

**
Biohacking generally refers to the practice of using science, technology, self-experimentation, and lifestyle interventions to “optimize” physical and cognitive performance, health, or longevity.

Originating in Silicon Valley and championed by tech entrepreneurs and fitness enthusiasts, biohacking encompasses a wide range of techniques—from nutritional tweaks and supplements to cutting-edge gene editing.

**Popular Techniques Among Biohackers:**

1.

**Dietary Approaches:**
   - **Intermittent Fasting:** Cycles of eating and fasting believed to promote weight loss and potentially improve metabolic health.

- **Ketogenic and “Bulletproof” Diets:** High-fat, low-carb regimes aimed at shifting metabolism and improving energy.

- **Gluten-Free, Dairy-Free, Anti-Inflammatory Diets:** Avoidance of foods thought to cause inflammation or disrupt the “gut.”
   - Scientific evidence for strict dietary regimes varies; intermittent fasting shows benefits in some trials, but restrictive diets can be unnecessary or harmful without medical need.

**Supplementation:**
   - **Vitamins, Nootropics, Probiotics, Adaptogens:** Widely used among biohackers to optimize energy, cognition, “gut health,” and reduce inflammation.

- The efficacy and safety of many supplements are unproven in large, controlled human studies; some may interact with medications or cause harm in excess.

**Physical Activity:**
   - **Extreme/Unique Exercise Regimes:** High-intensity interval training (HIIT), weightlifting, ice baths, and “habit stacking.”
   - Regular physical activity is broadly supported by scientific research, but extreme forms or overtraining may increase injury risk.

**Technology and Tracking:**
   - Use of wearables (Oura ring, continuous glucose monitors, heart rate variability trackers) for real-time health data, optimizing sleep, stress, and activity.

**Biohacking: Methods, Practices, and Supporting Evidence**

**What is Biohacking?

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Biohacking & Toxic Masculinity: How Women Dismantled The Boys Club

Author: Medical reviewer:
Beauty & Health Editor
By Jamie Schneider
Beauty & Health Editor
Jamie Schneider is the Beauty Editor at mindbodygreen. She has a B.A. in Organizational Studies and English from the University of Michigan, and her work has appeared in Coveteur, The Chill Times, and Wyld Skincare.
Medical review by
Bindiya Gandhi, M.D.
Physician
Dr. Bindiya Gandhi is an American Board Family Medicine–certified physician who completed her family medicine training at Georgia Regents University/Medical College of Georgia.
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Take a quick search for “biohacker” on TikTok (a tag with over 137 million views), and you’ll discover a slew of men discussing their muscle gains, testosterone levels, and cold plunge routines. You’ll see some women on the page, but the space is undoubtedly dominated by male energy. And think of the outspoken biohackers you may listen to on podcasts—chances are most of them are men, too.

This gendered approach to biohacking, while pervasive, is simply misguided. But now, women are asserting their authority in health optimization, and we expect their emphasis to soar in the year ahead.

Meet the experts

Molly Maloof. M.D.

Molly Maloof. M.D. provides health optimization and personalized medicine to high-achieving entrepreneurs, investors, and technology executives. Since 2012, she has worked as an advisor or consultant to more than 50 companies in the digital health, consumer health, and biotechnology industries. Her company, Adamo Bioscience, is working on sexual health optimization.

Gabrielle Lyon, D.O.

Gabrielle Lyon, D.O. is a board-certified family physician leading a disruption in modern medicine that focuses on the largest organ in the body: skeletal muscle. With a background that includes a combined research and clinical fellowship in geriatrics and nutritional sciences at Washington University and undergraduate training in nutritional sciences at the University of Illinois, she is a subject-matter expert on protein for health, performance, aging, and disease prevention.

Stacy Salvi

Stacy Salvi is the vice president of product and strategy at the medical device company Movano Health—the creator of the Evie Ring, the first medical-grade health wearable designed for women.

Mona Vand, PharmD

Mona Vand, PharmD studied pharmacy at Massachusetts College of Pharmacy and Health Sciences before developing her platform that shares holistic wellness advice.

Why biohacking isn’t just for “bros”

“Women are the original biohackers,” declares family physician, Molly Maloof, M.D. “Our hormones change throughout our lives, and this requires us to adapt accordingly.” Our biology means that women often need to constantly edit their health routines and experiment with lifestyle habits as time goes on. Isn’t that the very definition of biohacking?

Let's back up: Here at mindbodygreen, we think of “biohacking” as the practice of making lifestyle tweaks to optimize health and promote longevity. Not shortcuts—contrary to popular belief—but low-lift, everyday habits that complement and enhance an existing well-being routine. Think: optimizing your exercise regimen using tools and techniques (the “biohacks”) to enhance muscle growth. It’s essentially all about advancing health with “do it yourself” biology—a topic that can pique anyone’s interest, regardless of gender.

“[Women] have a most complex physiological lifecycle that requires us to track and manage fertility, pregnancy, breastfeeding, sexuality, perimenopause, and menopause. So we should be looking at biohacking as a path to even greater health and owning the opportunity to improve our lives,” says Maloof.

Yet many women feel intimidated to enter the biohacking scene as it exists today, turned off by the male-dominated energy in the room.

Women are the original biohackers

Molly Maloof, M.D

“Biohacking has always grabbed my attention, and I want to learn, but it sometimes makes me really stressed out, because I go into this state of perfectionism where I have to do all the things,” says doctor of pharmacy and wellness practitioner Mona Vand, PharmD. “The few biohackers that I followed were all males… it just doesn't really resonate with my energy.”

This male energy is not only frustrating but can potentially harm women’s health in the long run. After all, building muscle, prioritizing protein, and keeping an eye on various health metrics are crucial for longevity—in some cases, these proactive measures can save lives. When women don’t get involved, it can totally impact their well-being.

“Physical strength is nonnegotiable,” says functional medicine doctor Gabrielle Lyon, D.O. “I've been a physician since 2006, and I've never heard someone say, ‘I regret being physically strong.’” And the muscle-building space impacts more than just physiological changes, she says. “You optimize fertility, you make healthier babies, you help the epigenetics of your child,” Lyon adds. “You age better, your skin is better, you're less likely to fall and break a hip… There's nothing that doesn't get better the stronger you are.”

So let’s be clear: Dismantling the biohacking boy’s club is not just about helping women feel included; there are real health implications at stake here.

Why the biohacker space actually really needs a dose of feminine energy

I’d be remiss not to mention how biohacking culture can easily become synonymous with hustle culture (some experts don’t even like to use the term, as it implies you can achieve optimal health with quick fixes). But as more women enter the biohacking space, we expect it to become more intuitive and less about hitting every single benchmark. From there, we predict women will focus more on sustainable changes long-term instead of a gazillion quick hacks.

“I used to think it was a flex to do a million things at the same time and habit stack,” Vand recounts. “I now carve out time for doing things that have no efficiency to them, that are strictly for joy or rest and relaxation, versus benefiting my longevity or health. It can be a little bit obsessive when you get into this thought pattern, and I just don't resonate with it.”

This doesn’t mean biohacking will become “softer” as it embraces feminine energy. But there’s an opportunity here for women to introduce some much-needed flexibility into a space that has long felt needlessly rigid.

How the biohacker game is changing — and where it’s headed next

Thanks to a growing awareness that healthcare protocols should not be gendered, more women are now starting to pay attention to markers of performance like lean muscle, hormetic stressors, VO2 max, HRV, zone 2 training, etc. “Cold plunging, saunas, and harder, more physical activities are not just for the boys anymore,” adds Lyon—and we expect an even bigger shift to come in 2024.

“I think it’s super exciting to see biohacking become more mainstream and more women joining the conversation,” adds Maloof. She praises apps like Wild.AI, for women who aim to make lifestyle adjustments based on their menstrual cycle, and Zero, for women who want to dabble in time-restricted eating, both of which have helped to herald the shift.

Other existing health optimization companies are now inviting women into the conversation by introducing specific features informed by their unique physiology. WHOOP, for example, is currently working on upgrades to their popular health-tracking band: “We did a study recently where we learned a lot about how a woman's body changes during pregnancy and how her heart rate variability (HRV) will evolve. It'll decrease, and then about seven weeks before she gives birth, it will increase sharply,” explains Will Ahmed, WHOOP founder and CEO, on the mindbodygreen podcast

If a woman didn’t know that this was supposed to happen, she might look at this stark increase with a critical (or anxious) eye. “We've been able to take population-level data on pregnant women and now have a feature in the app, where a woman can see how her body is evolving alongside what other pregnant women are experiencing,” he adds.

Similarly, creatine supplements—from the ingredients to the packaging—are no longer marketed toward dudes looking to bulk up, and we are certainly here for it. After all, muscle mass is a predictor of longevity and healthspan, so supplements that promote it can help all of us “hack” our health in a big way.

And as Lyon notes, women have lower endogenous creatine stores compared to men (meaning, they have less available). And the amino acid has been associated with significantly increased muscle mass1 in both men and women, without increasing the risk of adverse outcomes2, like weight gain. “Most of the women in my practice will go on creatine, and they find anecdotally that it really improves both anaerobic and aerobic exercise performance,” Lyon shares.

Not to mention the supplement’s brain health perks: Taking creatine has been shown to benefit both working memory and processing speed3 as well as cognitive longevity4. “It is above and beyond simply sport performance,” adds Lyon, who has certainly seen an uptick in women curious about creatine.

As women feel intimidated (or even restricted) by biohacking spaces, some experts have taken it upon themselves to create new ones that center the feminine. For example, neuropsychophysiologist Kayla Osterhoff, MPH, offers a Women's Biohacking Mini Course, consisting of lifestyle routines and tips aligned with female physiology.

Osterhoff also recently presented at the 2023 Dragonfly Live Conference—among other female biohackers—to discuss the latest advancements in women's well-being and the “health revolution” that’s sweeping the mainstream. This conference aims to gather the “who's-who” in the health space to discuss cutting-edge strategies and innovations that optimize well-being—by having women biohackers at the forefront of the conversation, we can expect them to hit the main stage in a broader sense.

Or take the brand new Evie Ring, the first medical-grade health wearable designed for women, created by medical device company Movano Health. The technology ties hormonal information to a woman’s sleep, activity, and heart health data (among other metrics).

For instance, “If we notice a more restless night of sleep in the luteal phase, we'll point it out and also give some strategies for how somebody may attack their day, despite having not gotten as good of a night of sleep,” says Stacy Salvi, vice president of product and strategy at Movano Health. Similarly, “We know that there are cardiovascular implications when your estrogen drops, so they would want to keep a better look on the function of their heart,” she adds. “This would be something that they would be able to do from our app.”

Even the design itself was made with women in mind: The ring features an open band meant to accommodate how women’s fingers change in size throughout the day, so it can stay comfortable.

To avoid needless stress, Evie also limits the amount of goals you can set on the app. “You can only set three at a time, because we do not want to overwhelm [users] with a bunch of different things,” says Salvi. “I mean, I'm an overachiever. I probably would set every single goal and then fail at them.” And heaven knows we don’t need yet another way for busy, overwhelmed women to feel like they’re failing in a society with unsurmountable standards.

Forecasting the future

A woman walks up to the weight rack in 2024. She sees the usual suspects—bros pounding protein shakes and grunting through their bicep curls—but now she also sees a fair amount of women on the scene. They’ve got creatine supplements! Fitness trackers! Kettlebells! (And no, they aren’t pink.)

Granted, we still have a ways to go before women feel totally embraced by the biohacking space (we’d first have to close the centuries-long gender gap in science and medicine), but it’s clear that the arena is becoming way less of a boy’s club. “And the world is ready, my friend,” says Lyon. “The world is absolutely ready for this.”

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Biohacking generally refers to the practice of using science, technology, self-experimentation, and lifestyle interventions to “optimize” physical and cognitive performance, health, or longevity.

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Content: Home » biohacking, reprogramming mind and body to live better

biohacking, reprogramming mind and body to live better

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It’s called “biohacking” and it comes from Silicon Valley. Listen: The long life diet: the longevity formula is calorie restriction …

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It’s called “biohacking” and comes from Silicon Valley.

A relatively new word that comes from the union of “biology” and “hacker” and indicates the attempt to positively influence the functioning of the organism, by hacking, ie trying to “force”, one’s genetic heritage. A strategy to live better and longer launched years ago by Californian top managers and which then spread throughout the world. How to upgrade the “operating system” of the human body? No manipulation, here we are talking about “reprogramming” and new lifestyles. “Our genetic heritage has enormous unexpressed potential that can be activated,” he explains Fabio Piccinidoctor and psychoanalyst, researcher of Food Sciences of theMarche Polytechnic University, author of the book “Biohacking Techniques: how to optimize metabolism to increase health and longevity”. “Biohacking aims to activate epigenetic modifications by awakening dormant genes to enhance the body’s resources.” While genetics studies the structure of DNA, epigenetics the way genes express themselves, i.e. how much they are turned on or off. “We’re talking about a set of strategies that aim to strengthen the body from a physical and mental point of view, first of all through nutrition, sleep quality, physical activity, cognitive stimulation”. One of the pillars of biohacking and the intermittent fasting, Piccini adds, which plans to concentrate the diet in a precise time span of the day (to be followed always under medical supervision). «But it is equally important to take care of sleep, we know very well that sleep debt accelerates aging and that 30-40 percent of people sleep too little. Just as it is essential to check thehydration. Physical training must also be intermittent, with recovery periods in place to allow the muscle layers to rebuild. And then, you have to keep learning something at any age, another pillar of biohacking: studying music or a new language, for example, to counteract cognitive degeneration”.

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THE QUALITY OF LIFE

Sandra Milo is one of the supporters of this well-being and longevity strategy. «I’m lucky, at 90 I’m in full health and I still have a great desire to enjoy life. I don’t know if it’s a matter of genetics, nor do I think there is a “magic formula” of longevity, but I have certainly always been careful to lead a healthy life. There is nothing more beautiful than reaching old age with a smile and in full autonomy », says the actress who baptized the first Biohacking Suite in Rome. “Our mission is not so much to lengthen life but to improve its quality by lowering the rate of morbidity,” explains Dr. Claudio Urbani, specialist in regenerative medicine, neuroscience and stress therapy, head of the longevity center Umb. «Biohacking aims to modify dormant functional responses from non-use, from muscular or cardiovascular function to flexibility. It is about regaining possession of the body and the mind-body axis, starting with stress management». Diagnosing your own state of “aging” involves a series of tests. «The measurement of oxidative stress which measures the quantity of free radicals and indicates any inflammatory states, plethysmography to evaluate the functionality of the microcirculation of the legs, impedancemetry to study body composition, fat and lean mass, nutritional status and hydration level. And only after the check is a tailor-made program created for each person. Among the suggested therapies, there is also that of the cold which stimulates the metabolism and reduces inflammation ».

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Originating in Silicon Valley and championed by tech entrepreneurs and fitness enthusiasts, biohacking encompasses a wide range of techniques—from nutritional tweaks and supplements to cutting-edge gene editing.

**Popular Techniques Among Biohackers:**

1.

According to the studies, how does intermittent fasting help?

It reduces the risk of lifestyle-related diseases, such as metabolic disorders.

**Dietary Approaches:**
   - **Intermittent Fasting:** Cycles of eating and fasting believed to promote weight loss and potentially improve metabolic health.

Title:

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The baffling case of metabolically healthy obese people: Are they protected from chronic diseases?

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THE CONVERSATION

This article was originally published on The Conversation, an independent and nonprofit source of news, analysis and commentary from academic experts. Disclosure information is available on the original site.

——

Authors: Stan Kubow, Associate Professor, School of Human Nutrition, McGill University, McGill University and Michele Iskandar, Research Associate and Lecturer, School of Human Nutrition, McGill University

There is a growing global body positivity movement to combat the stigma and prejudice that overweight and obese individuals face. Some of this stigma arises from the perception that obese individuals are unhealthy.

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However, weight is only one of the metrics that medical professionals use to assess overall health. In fact, some studies have demonstrated that a significant number of obese people are metabolically healthy, leading to the contention that one could be healthy at any size. This phenomenon is referred to as metabolically healthy obesity (MHO). Research is now starting to explore what this means.

A person who is MHO has healthy blood pressure, normal levels of blood lipids (cholesterol and triglycerides) and normal blood sugar. Having high values of one or more of these measures significantly increases one’s risk for cardiovascular disease.

It has been proposed that metabolically healthy obese individuals might be protected against obesity-related diseases. The extent of this protection has been controversial within the scientific community, and this is partly because we still don’t have a standardized definition of MHO.

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So, over the past few years, researchers have gone to work to find out who can be defined as MHO, and to what extent people with MHO are protected from chronic disease.

Emerging consensus

Several large population studies have been initiated towards this end. So far, results show that, indeed, more rigorous definitions of MHO are needed. This can be achieved by including other measures of health such as insulin resistance and blood markers of inflammation. Insulin resistance is when the body does not respond well to the insulin hormone that helps to take up sugar from the bloodstream for use as a fuel for energy. This leads to elevated blood sugar and the consequent health complications.

An emerging consensus has been that although MHO individuals show some protection from chronic disease, they still seem to be significantly less protected than those who are metabolically healthy and lean.

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The fact that people with MHO still have a relatively higher disease risk led some researchers to suggest that the term “metabolically healthy obesity” could be a misnomer. Moreover, a majority of the MHO population tends to progress towards “metabolically unhealthy obesity” or MUO over a number of years, leading to enhanced risk of diabetes, cardiovascular disease and other disorders linked with obesity.

This leads to another question: is MHO truly protective over an entire lifetime, or is it just a question of time before MUO sets in, making MHO a transient state?

Protective factors

One factor to consider that might differentiate metabolically healthy versus unhealthy obesity is how fat is distributed in the body. A genetic predisposition for depositing fat under the skin, called subcutaneous fat, seems to play a protective role.

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People with this predisposition are mostly premenopausal women who accumulate subcutaneous body fat in the hips rather than the waist (pear-shaped). They are better protected against diabetes and cardiovascular disease compared to people whose body fat is located more in the abdomen (apple-shaped).

In contrast, obese individuals with a high waist circumference show excessive fat deposition in the abdomen and a pro-inflammatory state that leads to insulin resistance, which can be a precursor to Type 2 diabetes.

Researchers have also looked at the lifestyle habits that differentiate MHO from MUO individuals to see if development of metabolically unhealthy obesity can be prevented.

One factor is exercise. People with MHO are involved in regular physical activity to a greater extent than MUO individuals. The other is diet. Although dietary studies show mixed results, it seems that MHO people consume healthier diets such as the Mediterranean diet, which is rich in fish, fruits, vegetables, whole-grain products, legumes, extra virgin olive oil and nuts.

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Healthy diets provide anti-inflammatory and antioxidant benefits that help achieve a healthy metabolic profile. Indeed, of the MHO population, those who follow the Mediterranean diet seem to have lower mortality rates. Adopting these healthy lifestyle habits could help prevent the trajectory of metabolically healthy obesity moving towards metabolically unhealthy obesity over the course of a lifetime.

Is metabolically healthy obesity real?

So is MHO real and does it protect against disease? There isn’t a yes or no answer yet. The more we learn about it, the more nuance is injected into the concept that one can be perfectly healthy at any size. Based on the information we have so far, only a narrow segment of obese individuals are without risk of developing obesity-related chronic diseases.

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We also know a lot more about their characteristics. They have less abdominal fat and more subcutaneous fat. They have less insulin resistance, less inflammation and a metabolically healthy cardiovascular profile. They also have healthy lifestyle habits such as regular exercise and a healthy diet. It seems that genetics play a role as well.

We can expect more insight from major international population studies and clinical trials that are currently underway. But in the meantime, the age-old advice holds truer than ever: regular physical activity and a healthy diet are crucial to maintain health and longevity.

——

The authors do not work for, consult, own shares in or receive funding from any company or organisation that would benefit from this article, and have disclosed no relevant affiliations beyond their academic appointment.

——

This article is republished from The Conversation under a Creative Commons license. Disclosure information is available on the original site. Read the original article: https://theconversation.com/the-baffling-case-of-metabolically-healt https://theconversation.com/the-baffling-case-of-metabolically-

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According to the studies, how does intermittent fasting help?

It reduces the risk of lifestyle-related diseases, such as metabolic disorders.

- **Ketogenic and “Bulletproof” Diets:** High-fat, low-carb regimes aimed at shifting metabolism and improving energy.

- **Gluten-Free, Dairy-Free, Anti-Inflammatory Diets:** Avoidance of foods thought to cause inflammation or disrupt the “gut.”
   - Scientific evidence for strict dietary regimes varies; intermittent fasting shows benefits in some trials, but restrictive diets can be unnecessary or harmful without medical need.

Title:

Content: Introduction: Cell-Line Development Discussions at Biotech Week Boston

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The genetic sequence of the Chinese hamster ovary (CHO) cell line first was published just over a decade ago (1, 2). As with the Human Genome Project, genomic knowledge of that and other biopharmaceutical production cell lines — both animal and microbial — has expanded greatly alongside dramatic increases in computing power (3–5). Meanwhile new and improved genetic engineering techniques have offered the potential for targeted rather than random integration of transgenes into production cells. And regulators began to emphasize monoclonality more, demanding evidence that cell banks are derived from single engineered cells. All the while, cell-line development (CLD) groups faced increasing pressure to shorten timelines associated with their workflows.

As presentations at recent BPI conferences have revealed, “big data,” next-generation sequencing (NGS), automation, and powerful analytical technologies are helping biopharmaceutical companies achieve all those goals in parallel. And despite the widespread popularity of certain instruments, the talks at Biotech Week Boston (BWB) and the BPI Conference US West (BPI West) demonstrated that there is no one way to do so. Combining some of the approaches mentioned herein just might take biopharmaceutical production to new levels that not long ago would have been deemed impossible.

Below we highlight discussions at BWB. Look for more in our BPI West postevent report later this year.

Clonality and Transgene Integration
In September 2022, the BPI Conference program within BWB included both a preconference workshop on CLD and topical talks within the upstream production track. Presenters often spoke about how advanced gene-editing techniques can increase process productivity without compromising product quality. For example, biopharmaceutical companies increasingly are applying transposase-based gene editing to increase gene-integration targeting efficiency and accelerate CLD workflows (see the “Transposases” box). This is helping them reduce the number of subcloning/pool-screening steps that programs require.

Presenters highlighted the importance of high gene-copy numbers for optimized expression. Many talks highlighted the industry’s needs for technologies that enable continuous and/or real-time control of cell-culture parameters, especially for truly on-line process analytical technology (PAT). Many current systems need to improve interfaces between bioreactors and automated samplers/analyzers. BioPhorum representatives announced publication of a risk-assessment guide for raw materials used in CLD and other upstream processes. And some presenters called for greater attention to compositional chemistry, highlighting how such factors influence cell growth, consumption of media nutrients, and other key parameters.

In “Microfluidic-Chip–Based Single-Cell Cloning To Accelerate Biologic Production Timelines,” Geneva Alok (Amgen) explained that after genetic engineering and transfection, CLD can take ~28 weeks. That begins with initial selection of candidate clones through amplification by methotrexate (MTX) or methionine sulfoximine (MSX) and progresses through pool screening, subcloning, large-scale subcloning, clone screening at small production scales, and ultimately, large-scale screening. Alok is one of many speakers at BPI conferences in the past few years reporting on her company’s use of the Beacon instrument from Berkeley Lights to expedite selection and cloning steps. Loaded with thousands of clones, the machine photophoretically sorts single cells into NanoPen chip segments, where they are cultured and monitored continuously. (If two cells end up in a pen, the system can unpen and reload them; usually, the number of single-clone cultures is enough for conclusive results.) The system performs real-time analytics on those single-cell cultures. A complete cloning process takes about five days rather than the two to three weeks needed for fluorescence-activated cell sorting (FACS) methods. Even though the new technology is two to five times more expensive than the older one, large companies increasingly believe that its speed and throughput make up the difference. Amgen has replaced FACS-based screening with a Beacon instrument.

Alok highlighted that system’s Spotlight assay, which enables rapid screening to identify 10 highly productive clones (rather than the 96 productive clones typically identified by FACS). Monoclonality is documented by cell imaging, and CLD is accelerated by elimination of pool-screening steps. The Amgen team ran two different workflows to identify viable clones, then selected top candidates based on the results. When those findings were compared with results from fed-batch cultures of the same clones in Ambr microbioreactors, both performed comparably with similar stability based on parameters such as gene copy number and titer. Alok noted that the shortened workflow could generate clones with high genetic diversity after recovery, but that would require further study.

In “Cell-Bank Characterization and Retrospective Assessment of Clonality,” Guanghua Li (Merck) focused on test methods. Merck’s master cell bank (MCB) and working cell banks (WCBs) are produced according to good manufacturing practice (GMP) standards to reduce the possibility for introducing contamination. Li pointed out that no single testing regime can detect all potential contaminants (e.g., from culture media components). Cell-bank characterization typically entails assessment of a cell substrate’s identity, purity, and suitability for manufacturing applications. Such testing includes assays for viral contamination (often using minute virus of mice as a model) and in vitro/in vivo assays for adventitious agents (AAs).

Highlighting the revised ICH Q5A, which provides for application of NGS in AA testing, Li also summarized options for limit of in-vitro cell age (LIVCA) stability testing, noting that such testing usually is performed only once during cell banking. He noted that it is helpful to perform such testing at an intended production scale (e.g., 2,000 L) or simulation thereof. Doing so can help with study design and to demonstrate genotypic, phenotypic, and biosafety stability over the number of generations required for LIVCA testing. Li also summarized potential cloning strategies to use when demonstrating monoclonality, noting that adaptation to serum-free conditions (if applicable) should be performed before cloning steps are performed.

In “Use of Transposase-Mediated Integration To Reduce Screening and Shorten Cell-Line Development Timelines,” Hillary Warrington (Janssen) pointed to a primary pain point in CLD workflows: Clones can show variable success across transfections and preparations. “That needs to be optimized,” she said, adding, “Ideally, you want a high gene-copy number, which correlates positively with transfection success for expression titers.” Insertion efficiency for a gene of interest (GoI) can be increased in three ways, she said: through modification of transfection methods, gene-modification of host cells, and use of transposase mediators. She focused on the latter, describing her company’s application of ATUM Bio’s Leap-In transposase and transposon technologies.

The technology uses an RNA transposase to insert GoIs at different chromosomal sites and in high copy numbers — with more targeted genetic integration than a conventional plasmid system can offer. In Janssen’s study, ~95% of insertion sites used the transposon, and the resulting clones showed good stability with high copy numbers preserved across passages. The company’s historical data also show that application of Leap-In technology has increased monoclonal antibody (MAb) expression titers consistently since its adoption in 2018.

Warrington pointed out that less random integration enables CLD scientists to reduce the requisite number of subcloning steps, thus enabling earlier production of material for toxicology studies and other relevant activities. Moreover, subclones/populations do not require enrichment before sorting. Such benefits have helped Janssen to shorten its CLD timelines significantly by reducing clone-screening requirements. Other time savings have come from automation.

In “High-Throughput Characterization for IgM Cell-Line Development,” Christina Tsai (IGM Biosciences) reported on an accelerated testing approach. Increasingly, the sheer number of screening activities performed during CLD necessitates adoption of high-throughput instruments and methods. Tsai described her company’s collaboration with a technology supplier to improve and accelerate characterization studies for CLD activities toward production of IgM antibodies. Her company introduced PerkinElmer’s Lab Chip GX II system for high-throughput analysis of glycans, protein expression levels, and product characterization.

As a replacement for sodium-dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), the instrument performs a gel-electrophoretic method on a multiuse microfluidic chip: sodium dodecylsulfate microcapillary electrophoresis (µCE-SDS). Requiring at least 2.5 µL of material, the instrument can analyze each sample in about one minute, and a chip can be reused for 400 samples. IGM’s workflow requires about 1.5 hours of preparation, 50 minutes to run 50 samples, and under an hour for analysis. Tsai highlighted that the method can be performed 6× faster than affinity high-performance liquid chromatography (HPLC) and 25× faster than size-exclusion chromatography (SEC). The system also can be used to assess titer levels and glycan profiles 50× faster than a system for hydrophilic-interaction liquid chromatography (HILIC) testing.

In “Growing Pains: The Ups and Downs of Building a CLD Platform Process from Scratch,” Allyn Spear (Elanco) chronicled successes and pitfalls from development of a host-cell line for production of a canine antibody product. Before 2000, he explained, biologics equated to vaccines in animal health — but that veterinary companies now are expanding into other modalities. Elanco is among the animal-health companies developing capabilities for biological products such as antibodies and other proteins.

Tasked with identifying a glutamine synthetase (GS)–knock-out host-cell line to express a canine antibody, Spear and his colleagues also needed to identify optimal culture parameters toward a long-term goal of producing that antibody at commercial scale. First, they experimented with the GS piggyBac and Leap-In transposon systems (from Lonza and ATUM, respectively) to identify an effective vector strategy. Elanco settled on a two-plasmid transfection approach but found significant variability in transfection efficiency and posttransfection viability. With too many variables involved to narrow down the root cause, the team needed to apply additional reagents to increase transfection consistency.

They used the Berkeley Lights Beacon system for early clone selection activities. Spear noted, however, that the project required custom reagent development because it involved nonhuman antibodies. The team used Ambr microbioreactors from Sartorius for final clone screening. He highlighted difficulties in CLD for start-ups and other small teams: e.g., supply chain problems with equipment availability and inconsistent quality across consumables from different suppliers.

In “Accelerating Cell-Line Development Processes Through Automation Platform Integration,” Woo Chi (Just–Evotec) explained that his company makes abundant use of automation technologies because of a preference for continuous biomanufacturing. He noted that his CLD team faced difficulties in implementing a robotic platform for cell transfer (passaging/retention in a perfusion process). Initially, the system was transferring only 10% of cells, but the team adjusted software scripts over several programming iterations to increase cell transfer to 90%.

In “Implementing Beacon Optofluidic Technology for Accelerated Cell-Line Development for Complex Subunit Vaccine Candidates,” Jishna Ganguly (GSK) pointed out that most reported applications of Beacon technology have been in antibody process development. GSK is the first company to use the photophoretic technology to accelerate CLD activities in production of antigens used in a subunit vaccine.
Ganguly shared a feasibility study that her company used in adopting the system. GSK previously used a 12-day workflow that combined application of a Cell Metric cell imager (Solentim) and an assay from Luminex. The Beacon system accelerated CLD considerably while generating results comparable to those from the previous workflow.

Ganguly concluded by highlighting continuing CLD needs of vaccine makers, who could benefit from solutions that reduce manual handling during testing and that minimize the number of instruments required for the many kinds of screening performed (e.g., for clone productivity, product quality, monoclonality). Although many choices are available, drug makers often apply many different technologies, increasing experimental effort and time along with the complexity of system integration.

In “Evaluation of Transposase Technology To Generate Highly Productive CHO Cell Lines,” Eric Lee (Biogen) explained that his company, like many other drug developers, is leveraging transposase-mediated gene editing to expedite CLD activities. His team accelerated CLD by two weeks through applying Lonza’s GS piggyBac transposon system and eliminating steps for MTX amplification. That allowed the program to move right into evaluation of material from recovered pools. Transfection recoveries were good, and although the transposon system generated considerably more random integration than Biogen’s legacy method did, pool-expressed material was comparable in quality, with aggregation and isoform levels that aligned with those from historical data. In subsequent studies, Biogen determined that the transposase system also was compatible with MTX amplification and that adding that step back into the workflow increased expression titers significantly. Ultimately, the company improved its CLD workflow to generate material for toxicology studies five months earlier than would have been possible with the legacy method.

In “Epigenetic Comparison of CHO Hosts and Clones Reveals Divergent Methylation and Transcription Patterns Across Lineages,” Meiping Chang (Merck) noted that genomic and epigenomic analyses are becoming critical to assessment of transgene integration and clone selection. The trick, she explained, is to evaluate epigenetic regulators of transcription without dismantling host-cell DNA, especially because epigenetic markers can be inserted or removed.
She explained that DNA-methylation patterns are passed down by parent cells, although new patterns can emerge during cloning. Underlying mechanisms of such changes include differential expression of epigenetic readers, erasers, and effectors. To demonstrate, Chang shared her team’s testing of two host-cell lines for genomic changes over several generations by searching for differentially methylated regions (DMRs). Most clonal cells showed inherited changes in methylation patterns. That, Chang explained, could indicate development of a trait for long-term genetic stability. Using such information could help CLD scientists to find ideal candidates for transgene integration.

Emphasis on Speed
The focused CLD workshop at Biotech Week Boston culminated in a panel discussion titled “Techniques and Technologies To Accelerate CLD and Solve Bottleneck Challenges,” with Allyn Spear, Woo Chi, Hillary Warrington, and William Rice (Nexcelom Bioscience). Panelists noted that companies increasingly are using artificial intelligence (AI) to identify and optimize vector constructs. Both on the stage and in the audience, there was particular excitement for growth in libraries for human antibodies. One attendee asked whether it could be helpful to use a reporter gene during early CLD activities. Panelists explained that such genes are not secreted as antibodies are; thus, they might be useful for gauging transfection efficiency but not for expression titer.

Many participants questioned whether to use random or targeted gene integration during CLD. Panelists explained that pools from clones generated using the former technique often show more heterogeneity and experience epigenetic drift over time. Technologies for targeted integration can generate more homogenous pools that might show less drift. However, even minor epigenetic drift must be characterized, so drug makers using targeted integration would not be relieved of some associated regulatory burden for product characterization. One panelist noted that current technologies for targeted gene integration also result in some random integration and that such occurrences could cause regulatory questions. Considering the significant costs associated with targeted-integration technologies, developers should evaluate their needs carefully to help ensure that such solutions will be worth the investment.

Transposases
First identified by Barbara McClintock in the mid-20th century (work that earned her the 1983 Nobel Prize in physiology/medicine), transposase enzymes bind to the end of a transposon and catalyze its movement to another part of a cell’s genome, typically through a cut-and-paste or replicative mechanism. In nature, transposition is both useful for evolution and adaptation to changing environments and potentially dangerous to individual survival. Because genomic instability can lead to unhelpful mutations, most such genes are down-regulated. But transposases have great utility in genetic engineering.Further Reading
Aziz RK, Breitbart M, Edwards RA. Transposases Are the Most Abundant, Most Ubiquitous Genes in Nature. Nucl. Acids Res. 38(13) 2010: 4207–4217; https://doi.org/10.1093/nar/gkq140.Grabundzija I, et al. Comparative Analysis of Transposable Element Vector Systems in Human Cells. Molec. Ther. 18(6) 2010: 1200–1209; https://doi.org/10.1038/mt.2010.47.Gustafsson C, et al. Engineering Genes for Predictable Protein Expression. Prot. Expr. Purif. 83(1) 2012: 37–46; https://doi.org/10.1016/j.pep.2012.02.013.Rajendra Y, Peery RB, Barnard GC. Generation of Stable Chinese Hamster Ovary Pools Yielding Antibody Titers of Up to 7.6 g/L Using the PiggyBac Transposon System. Biotechnol. Prog. 32(5) 2016: 1301–1307; https://doi.org/10.1002/btpr.2307.

Vics Z, Izsvák Z. A Whole Lotta Jumpin’ Goin’ On: New Transposon Tools for Vertebrate Functional Genomics. Trends Genet. 21(1) 2005: 8–11; https://doi.org/10.1016/j.tig.2004.11.008.

Cell-Line Engineering at BPI West
Held on 27 February through 2 March 2022, the BPI West program in San Diego, CA, included a full track devoted to cell-line development. Many themes followed from the BWB discussions a few months before, but the overall emphasis shifted toward applying advanced genetic engineering techniques to improve cell stability, productivity, predictability, and viability. Technological potentialities touted over recent years seem to be coming to fruition in big ways in the post-COVID era. Look for a full report on the program coming up later this year.

References
1 Wurm F, Hacker D. First CHO Genome. Nat. Biotechnol. 29, 2011: 718–720; https://doi.org/10.1038/nbt.1943.

2 Xu X, et al. The Genomic Sequence of the Chinese Hamster Ovary (CHO)-K1 Cell Line. Nat. Biotechnol. 29, 2011: 735–741; https://doi.org/10.1038/nbt.1932.

3 Lewis N, et al. Genomic Landscapes of Chinese Hamster Ovary Cell Lines As Revealed By the Cricetulus griseus Draft Genome. Nat. Biotechnol. 31, 759–765 (2013); https://doi.org/10.1038/nbt.2624.

4 Rupp O, et al. A Reference Genome of the Chinese Hamster Based on a Hybrid Assembly Strategy. Biotechnol. Bioeng. 115(8) 2018: 2087–2100; https://doi.org/10.1002/bit.26722.

5 Lin D, et al. CHOmics: A Web-Based Tool for Multi-Omics Data Analysis and Interactive Visualization in CHO Cell Lines. PLOS Comput. Biol. 22 December 2020; https://doi.org/10.1371/journal.pcbi.1008498.

Brian Gazaille is managing editor, and corresponding author Cheryl Scott is cofounder and senior technical editor of BioProcess International (part of Informa Connect Life Sciences); 1-212-600-3429; cheryl.scott@informa.com; https://www.bioprocessintl.com.

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Content: She's fighting to stop the brain disease that killed her mother before it gets her.

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CAMBRIDGE, Massachusetts ‒ Sonia Vallabh watched helplessly as her 51-year-old mother rapidly descended into dementia and died. It didn't take long for Vallabh to realize she was destined for the same rare genetic fate.

Vallabh and her husband did what anyone would want to do in their situation: They decided to fight.

Armed with little more than incredible intellect and determination they set out to conquer her destiny.

A dozen years later, they've taken a major step in that direction, finding a way to shut off enough genetic signals to hold off the disease.

And in the process of trying to rescue Vallabh, they may save many, many others as well.

In a paper published Thursday in the prestigious journal Science, Vallabh and her husband, Eric Minikel, and their co-authors offer a way to disrupt brain diseases like the one that killed her mother.

The same approach should also work against diseases such as Huntington's, Parkinson's, ALS and even Alzheimer's, which result from the accumulation of toxic proteins. If it works as well as they think, it could also be useful against a vast array of other diseases that can be treated by shutting off genes.

"It doesn't have to be the brain. It could be the muscles. It could be the kidneys. It could be really anywhere in the body where we have not easily been able to do these things before," said Dr. Kiran Musunuru, a cardiologist and geneticist at the University of Pennsylvania's Perelman School of Medicine, who wasn't involved in the research but wrote a perspective accompanying the paper.

So far, they've proven it only in mice.

"The data are good as far as they go," Vallabh said this week from her office at the Broad Institute of Harvard and MIT, where she has worked since getting a PhD at Harvard. She had already gotten a law degree from the university, but she and Minikel, then a transportation planner, both pursued biology degrees after her mother's death. Now, they work together at the Broad.

"We're far from this being a drug," Vallabh said. "There's always, always reason for caution. Sadly, everything is always more likely to fail than succeed.

"But there is justifiable reason for optimism."

A terrible disease

The disease that killed Vallabh's mother was one of a group of conditions called prion diseases. These include mad cow disease, which affects mostly cattle, scrapie, which affects sheep, and Creutzfeldt-Jakob disease, which kills about 350 Americans a year ‒ most within months of their first symptom.

These diseases are triggered when the prion protein found in all normal brains starts misfolding for some reason, as yet unknown.

"Prion disease can strike anybody," Vallabh said, noting the 1 in 6,000 risk to the general population.

Though prion diseases are, in some cases, contagious, a federal study earlier this year concluded that chronic wasting disease, found in deer, elk and moose, is very unlikely to pass to people who eat the meat of sick animals.

In Vallabh's case, the cause is genetic. Vallabh discovered after her mother's death that she carries the same variant of the same gene that caused her mother's disease, meaning she will certainly develop it.

The only question is when.

"The age of onset is extremely unpredictable," Vallabh said. "Your parent's age of onset doesn't actually predict anything."

How the gene-editing tool works

Vallabh, Minikel and their colleagues developed a gene-editing approach they call CHARM (for Coupled Histone tail Autoinhibition Release of Methyltransferase).

While previous gene-editing tools have been described as scissors or erasers, Musunuru described CHARM as a volume control, allowing scientists to tune a gene up or down. It has three advantages over previous strategies, he said.

The device is tiny, so it fits easily inside the virus needed to deliver it. Other gene-editing tools, like CRISPR, are bigger, which means they need to be broken into pieces and much more of the virus is needed to deliver those pieces to the brain, risking a dangerous immune reaction.

CHARM, Musunuru said, is "easier to deliver to hard-to-deliver spaces like the brain."

At least in the mouse, it also seems to have reached throughout the brain, making the desired genetic change without other, unwanted ones, Musunuru said.

And finally, the research team figured out a way to turn the gene editor off after its work was done. "If it's sticking around, there's the potential for genetic mischief," Musunuru said.

One shot on goal

While researchers, including Vallabh, continue to work to perfect an approach, the clock for Vallabh and others is ticking.

Right now there's no viable treatment and if it takes too long to develop one, Vallabh will miss her window. Once the disease process starts, like a runaway train, it'll be much harder to stop than it would be to just shut the gene off in the first place.

The more of the prion protein in the brain, the more likely it is to misfold. And the more likely it is for the disease to spread, a process that co-opts the natural form of the protein and converts it to the toxic form.

That's why getting rid of as much of it as possible makes sense, said Jonathan Weissman, the senior author on the study, who leads a lab at the Whitehead Institute, a biomedical research institute next to the Broad.

"The biology is really clear. The need (for a cure) is so compelling," Weissman said.

Every cell in the brain has the gene for making the prion protein. By silencing even 50% of those genes, Weissman figures he can prevent the disease. In mice, CHARM silenced up to 80% to 90%.

"We've figured out what to deliver. Now we have to figure out how to deliver it," he said.

Another of the paper's co-authors, the Broad's Ben Deverman, published a study late last year showing he could deliver a gene-therapy-carrying virus throughout the brain. Others are developing other viral delivery systems.

And Vallabh and Minikel have hedged their bets, helping to develop a so-called antisense oligonucleotide, or ASO, which uses another path for stopping the gene from making the prion protein.

The ASO, which is in early trials in people by a company called Ionis Pharmaceuticals, requires regular treatment rather than the one-and-done of a gene therapy. Recruitment for that trial had to be paused in April because the number of would-be volunteers outstripped the available slots.

Personalized therapy: This lifesaving treatment was designed for one. Could it be the future of medical care?

Vallabh isn't ready yet to start any treatment yet herself.

"She has one shot on goal," Musunuru said. "At some point, she'll have to decide what's the best strategy."

In the meantime, the clock Vallabh can't see continues to tick toward onset.

She and Minikel stay exceedingly busy with their research along with their daughter, almost 7, and 4-year-old son ‒ both born via IVF and preimplantation genetic testing to ensure they wouldn't inherit her genetic curse. (They were super lucky, Vallabh notes, to be living in Massachusetts where IVF is at least "approachable" financially.)

"There is a mountain ahead of us," Vallabh said of the path to a cure. "There's still a lot of hurdles, there's still a lot to figure out."

Karen Weintraub can be reached at kweintraub@usatoday.com.

This article originally appeared on USA TODAY: After dementia killed her mother a medical researcher looks for a cure

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**Summary of Criticism and Controversy Around Hagia Sophia's Conversion and Broader Context:**

- **Religious and Interfaith Concerns:**
  - The reconversion of Hagia Sophia from a museum to a mosque in 2020, and a similar subsequent conversion of the Chora Church (another Byzantine monument), has sparked controversy, especially among Christian communities and international observers.

- Some within the Orthodox Christian community have expressed disappointment, noting that while they cannot venerate these sites as they once did, they seek areas of unity with Muslims and other faiths.

- **National and Political Implications:**
  - Many Turks, particularly Muslim worshippers, see the restoration of Hagia Sophia and other former Byzantine churches to their prior status as mosques as a source of national pride and a reclaiming of Ottoman and Islamic heritage.

This move is widely interpreted as a political victory by Turkish President Recep Tayyip Erdogan, reinforcing his image as a protector of Islamic values and Turkish sovereignty.

- Critics argue that these actions are politically motivated, serving to rally Erdogan’s conservative base and shift attention from domestic challenges by stirring nationalist and religious sentiment.

- The moves have raised concerns among secular Turks and religious minorities (such as the Greek Orthodox community in Turkey) about increasing religious undertones in Turkish politics and possible challenges to multicultural and multi-faith coexistence.

- **International and UNESCO Reactions:**
  - The reconversion contravenes the spirit of Hagia Sophia’s status as a UNESCO World Heritage Site, which is meant to be accessible and respected as a symbol of global human heritage.

- UNESCO and international leaders have raised concerns regarding ongoing preservation and interfaith accessibility at these historic monuments.

**Summary of Criticism and Controversy Around Hagia Sophia's Conversion and Broader Context:**

- **Religious and Interfaith Concerns:**
  - The reconversion of Hagia Sophia from a museum to a mosque in 2020, and a similar subsequent conversion of the Chora Church (another Byzantine monument), has sparked controversy, especially among Christian communities and international observers.

Title:

Content: After Hagia Sophia, Turkey converts a second ancient Byzantine church into a mosque

Hundreds of worshippers filled the narrow streets of Istanbul's Fatih district on Friday to hear the sounds of Friday prayers ringing out from the newly inaugurated Kariye Mosque.

Once known as the Chora Church, the site spent the last 79 years as a museum. But it is now the latest structure to be converted back into a mosque by the government of Recep Tayyip Erdoğan, following the conversion of Hagia Sophia in 2020. While viewed as a triumph by many Turkish Muslims, the Greek Orthodox Church has decried it as an "ill-advised decision" that "makes a mockery of the Turkish government's commitment to religious tolerance and religious freedom."

Chora's conversion was announced shortly after Hagia Sophia's, but its opening for prayer was delayed by years of restoration work.

The showing for the first Friday prayers at Kariye was relatively quiet compared to Hagia Sophia, which drew hundreds of thousands who filled the streets with prayer rugs for blocks and blocks away from the building. By contrast, the crowd on Friday was relatively average for a mosque in one of Istanbul's more religiously conservative neighborhoods, where Friday worshippers frequently spill out onto the streets.

Both the Chora Church and Hagia Sophia are 4th-century Byzantine constructions. They spent nearly a millennium as Christian holy sites before being converted to mosques by the Ottomans after their conquest of Constantinople in the 15th century. Despite their conversions, they remained revered by Orthodox Christians. After the founding of a secular Turkish Republic in the early 20th century, the decision was made for both to be neither mosque nor church but simply museums.

"Both Hagia Sophia and Chora embodied Byzantine and Ottoman pasts, and they became symbols of co-existence and multi-faith living practices. Their conversion fundamentally implies a hierarchy, prioritizing the Islamic past over all other layers, Greek Orthodox, Armenian, Jewish, Syriac, et al," Dr. Özgür Kaymak, a lecturer and researcher on minority rights at Istanbul's MEF University, told Religion News Service in an email.

Rededicating the sites as mosques is more than just a status change. According to Islamic tradition, artwork of human figures are forbidden in places of prayer, while in Orthodox Churches, icons of Christ, saints and other biblical figures are a defining feature.

In Hagia Sophia, the mosaics of Jesus and Mary have been covered with curtains since the conversion. While Erdoğan promised Hagia Sophia would remain fully accessible and free to all visitors, Turkey reneged on that earlier this year, reimposing an entrance fee on tourists and relegating them to the upper level, while the main floor is exclusive to Muslim worshippers.

Observers have also criticized the Turkish Directorate of Religion's caretaking of the site, noting damage that did not exist when it was under the purview of the Ministry of Culture and Tourism.

Though Chora is significantly smaller than Hagia Sophia, it is home to some of the best-preserved examples of late-Byzantine art, including mosaics, icons and frescoes, and is considered a UNESCO World Heritage site.

"For us it's a very important place because it remembers our culture and our traditions here," Father Evangelos Markantonis, an Orthodox priest who was leading a group of theology students to Chora on Friday, told Religion News Service.

"Though we cannot venerate as Orthodox Christians, we have to try to find things we can be united on. Only with dialogue and good deeds can we continue our lives," Markantonis, who is also a professor at the University of Athens, said when asked about the controversy.

Erdoğan had long refused calls from his right to convert churches like Hagia Sophia and Chora, telling supporters in 2018 to fill the nearby Sultan Ahmet Mosque (also known in English as the Blue Mosque) before they talked about needing to pray in Hagia Sophia.

Nonetheless, he made an about-turn in 2020.

"My dear nation, the conquest of Istanbul and the conversion of the Hagia Sophia into a Mosque are among the most glorious chapters in Turkish history," Erdoğan said at the time in a speech inaugurating the Hagia Sophia as a mosque.

"This is the most honorable day that Islam has been looking forward to, Greek Constantinople has become Turkish Istanbul," he added, quoting the Turkish poet Nazim Hikmet.

"The resurrection of Hagia Sophia heralds the liberation of the al-Aqsa Mosque. The resurrection of Hagia Sophia is the footsteps of the will of Muslims across the world to come out of the interregnum," Erdoğan said in his concluding remarks.

Chora's opening as the Kariye Mosque comes just weeks after Erdoğan's Justice and Development Party (AKP) suffered its largest electoral defeat in two decades during Turkey's nationwide municipal elections.

"The recent conversion of Chora into a mosque may be due to Erdoğan's many problems, including his geopolitical power politics, his appeal to religio-nationalism to revive his electoral popularity after the March elections or his tactic to divert attention from Turkey's economic downturn," Kaymak said.

Many analysts have attributed the loss to AKP bleeding votes to the more Islamist-leaning New Welfare Party, which has pressured Erdogan over his response to the Israel-Hamas war and humanitarian situation in the Gaza Strip.

Muslim worshippers at the mosque on Friday said they viewed its rededication as an Islamic site to be a national win for Turkey.

"This mosque is one of the symbols of the Conquest. We are proud and happy. There are very beautiful frescoes inside. They were also nicely renovated before they were closed. May it be good for our country and our nation," a bystander, Ahmet Öteyüzoğlu, told Turkish media.

"Thank God for these days. Not everyone has the opportunity in their lifetime," another bystander named Mehmet Çelik told the Turkish news service Anadolu Agency, in reference to having been able to pray at both the rededicated Hagia Sophia and Kariye Mosques.

However, Greece, the U.S. State Department and Orthodox Christian institutions worldwide have all criticized the decision to revert the structure into a mosque.

"I want to publicly express my intense dissatisfaction, interpreting the feelings of all Greek women and men, for the completely unnecessary conversion of a historic Byzantine temple, the Monastery of Chora, into a mosque. It is, I believe, an action that offends the rich history of Istanbul itself as a crossroads of cultures," Greek Prime Minister Kyriakos Mitsotakis said shortly before leaving for Turkey for a planned meeting with Erdogan.

"We encourage the Turkish government to preserve and ensure access to sites and buildings that have hosted different religious communities in a way that respects their diverse histories," a State Department spokesperson said in response to questions from Greek media.

"The Turkish government's appropriation of both as the property of one faith group not only constitutes yet another sign of that government's contempt for Turkey's rich Orthodox Christian heritage but further imperils the religious freedom of the Ecumenical Patriarchate and the remaining Christians of that land," the Archons of the Ecumenical Patriarchate said in a statement.

© Religion News Service

How did Muslims and Orthodox Christians react to the reconversion?

Who was angered by the reconversion of Hagia Sophia in July 2020?

- Some within the Orthodox Christian community have expressed disappointment, noting that while they cannot venerate these sites as they once did, they seek areas of unity with Muslims and other faiths.

Who was angered by the reconversion of Hagia Sophia in July 2020?

- **National and Political Implications:**
  - Many Turks, particularly Muslim worshippers, see the restoration of Hagia Sophia and other former Byzantine churches to their prior status as mosques as a source of national pride and a reclaiming of Ottoman and Islamic heritage.

How did Muslims and Orthodox Christians react to the reconversion?

How did the role of Hagia Sophia change following the Ottoman conquest?

Hagia Sophia became a central Islamic religious site for centuries

What kind of domestic reaction did Erdogan's reconversion of Hagia Sophia receive?

What political motivations influenced Erdogan's decision to reconvert Hagia Sophia?

Erdogan's decision was influenced by Turkey's Islamic resurgence.

This move is widely interpreted as a political victory by Turkish President Recep Tayyip Erdogan, reinforcing his image as a protector of Islamic values and Turkish sovereignty.

What kind of domestic reaction did Erdogan's reconversion of Hagia Sophia receive?

What political motivations influenced Erdogan's decision to reconvert Hagia Sophia?

Erdogan's decision was influenced by Turkey's Islamic resurgence.

- Critics argue that these actions are politically motivated, serving to rally Erdogan’s conservative base and shift attention from domestic challenges by stirring nationalist and religious sentiment.

Title:

Content: Hindustantimes wants to start sending you push notifications. Click allow to subscribe

Turkey's ancient ruins: Explore rich past of Temple of Artemis, Troy and more

Updated on Jan 08, 2023 06:36 PM IST

In recent years, more and more people have been drawn to the ruins of Turkey, eager to explore the country’s rich past. The ruins of Turkey are some of the most impressive in the world. Visit these best-preserved archaeological sites on your next travel

By | Edited by Zarafshan Shiraz, Delhi

The Ancient Ruins of Turkey have been a source of fascination for travellers for centuries. From the majestic city of Istanbul to the ancient ruins of Ephesus, Turkey is a country that is full of history and culture. In recent years, more and more people have been drawn to the ruins of Turkey, eager to explore the country’s rich past.

The ruins of Turkey are some of the most impressive in the world. The ancient city of Ephesus is one of the best-preserved archaeological sites in the world, and it has been a popular destination for tourists for many years. The ruins of the city are a testament to the power and wealth of the ancient city. The city was once home to the Temple of Artemis, one of the Seven Wonders of the Ancient World. The ruins of the temple are still visible today and are a popular attraction for tourists.

Other ancient ruins in Turkey include the ruins of Troy, the ancient city of Hattusa, and the ruins of Aphrodisias. Troy was the site of the legendary Trojan War, and the ruins of the city are still visible today. The ruins of Hattusa are the remains of the capital of the Hittite Empire, and they are an impressive sight. The ruins of Aphrodisias are the remains of an ancient Greek city, and they are also a popular tourist attraction.

Exploring the ancient ruins of Turkey can be a very rewarding experience. Many of the ruins are open to the public, and visitors can explore them at their own pace. Visitors can also take guided tours of the sites, which can provide valuable insight into the history and culture of the country.

The ruins of Turkey are also a great place to learn about the country’s ancient history. Many of the ruins are home to artifacts and relics from the past, and visitors can learn about the culture and history of the country by examining these artifacts. The ruins of Troy, for example, are home to many artifacts that were used in the Trojan War.

Exploring the ancient ruins of Turkey is also a great way to experience the country’s culture and traditions. Many of the ruins are home to traditional Turkish architecture, and visitors can learn about the country’s traditional customs by visiting these sites. Visitors can also experience traditional Turkish cuisine by visiting the many restaurants and cafes that are located near the ruins.

Exploring the ancient ruins of Turkey is a great way to learn about the country’s past and experience its culture. The ruins are a great way to experience the country’s history and culture, and they are a great way to get an insight into the country’s past. Visiting the ruins of Turkey is an experience that will stay with you for a lifetime.

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What political motivations influenced Erdogan's decision to reconvert Hagia Sophia?

Erdogan's decision was influenced by Turkey's Islamic resurgence.

- The moves have raised concerns among secular Turks and religious minorities (such as the Greek Orthodox community in Turkey) about increasing religious undertones in Turkish politics and possible challenges to multicultural and multi-faith coexistence.

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December 9, 2021

Christian leaders look forward to Turkey's new law on religious foundations

This is an issue of vital importance for local Christian communities whose places of worship are entrusted to and managed by their governing bodies

Religious minorities in Turkey are looking forward to the increasingly Islamo-nationalist government making good its promise of outreach to non-Muslims by giving them freedom to establish and run their religious and social institutions.

The Turkish government is preparing to issue a new regulation on the administrative management of foundations linked to non-Muslim faith communities, which have been paralyzed for years by a legislative impasse that blocks the renewal of their governing bodies.

This is an issue of vital importance for local Christian communities such as Armenians and Assyrians, whose places of worship – many of them historic --, other real estate and public institutions are entrusted to and managed by foundations.

Burhan Ersoy, head of the state-run Directorate of the Foundations, has confirmed that the plan for the drafting of a new regulation concerning the elections for the assignment of offices within the foundations has come into being, Fides reported.

The draft of the text has been drawn up, taking into account requests and proposals from the various minority communities, and it is expected that the new rules, after obtaining government approval, would be implemented by April 2022.

The previous electoral regulation for the top management of foundations had been suspended in 2013, after the government chose to establish new procedures to make the management of real estate more functional and transparent.

This led to some confusion with representatives of the various minority communities unable to come to a consensus to either accept or reject the new electoral rules, Ersoy said.

The Syrian Orthodox Christian Süleyman Can Ustabaşı, current representative of the non-Muslim Foundations within the Assembly of Foundations, has said that the structure of foundations and their membership profiles had changed over time.

The Turkish government already had consultations with the representatives of religious minorities to resolve the dispute over board elections for their foundations.

But Ustabaşı now urges Turkish authorities to carry out another consultation with representatives of minority communities before the new text is to be implemented in 2022.

The legal status of the Foundations is still based on the Peace Treaty of Lausanne, signed in 1923 by Turkey and the Entente powers (British Empire, France and Russian Empire) victorious from the First World War.

The treaty guaranteed equality before the laws and the freedom to promote and manage "religious and social institutions" for non-Muslim communities in Turkey.

However, the state had seized properties run by these foundations and had to contend with litigation as these foundations claimed the rights guaranteed by the Treaty of Lausanne.

Also, starting from 1936, legislative provisions had allowed foundations of non-Muslim communities to acquire new properties. But in 1974, this guarantee was canceled and the state begun seizing assets acquired by these foundations.

Between 2013 and 2018 about 1,084 properties were returned to the foundations linked to non-Muslim communities, and 20 places of worship restored and given back to the foundations, according to official data.

Greek Orthodox Laki Vingas, in a speech published by the Armenian-Turkish newspaper Agos, said that the legislative deadlock on the question of the regulation of the foundations has had a negative effect on the life of the different ecclesial communities.

The block in the processes of renewal of the governing bodies of foundations had led to the expulsion of young people from the institutions linked to their communities, and also that many voluntary activities were "unfortunately interrupted".

Non-Muslim places of worship

Within the general Assembly of the Turkish Foundations are some 167 minority foundations. The Turkish Greek community has 77, the highest number of foundations. The Armenian community has 54 and the Jewish community has 19 foundations.

Turkish President Recep Tayyip Erdogan after a cabinet meeting held on October 25 announced that authorities had put on the agenda the question of the elections of the directorates of the foundations linked to religious minorities.

He also spoke of fundamental tools for the management of assets and resources destined for non-Muslim places of worship.

The issue of the foundations come at a time when Turkish authorities have expelled more and more Protestant Christians -- foreigners and Turkish converts-- on grounds they constitute a "threat to national security".

Turkey had also recently transformed museums into mosques, such as the Hagia Sophia and the Church of the Holy Saviour in Chora.

Both churches had been turned into museums after the end of the Ottoman Empire and the establishment of modern Turkey.

The Church of the Holy Saviour in Chora was built by the Byzantines in the 5th century. The Ottomans then turned it into a mosque after they captured Constantinople in 1453. And it remained a Muslim worship space until the end of the Second World War when it was transformed into a museum.

The same happened to the Hagia Sophia Museum in Iznik, and the high-profile reconversion of the Hagia Sophia Museum, Istanbul's most identifiable monument.

Many observers believe that President Erdogan, weakened internally by a bad economy and a lack of political perspective, is compensating with erasing part of the country's multiculturalism, aimed at shoring up the ranks of his Islamo-nationalist base.

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What political motivations influenced Erdogan's decision to reconvert Hagia Sophia?

Erdogan's decision was influenced by Turkey's Islamic resurgence.

Who was angered by the reconversion of Hagia Sophia in July 2020?

- **International and UNESCO Reactions:**
  - The reconversion contravenes the spirit of Hagia Sophia’s status as a UNESCO World Heritage Site, which is meant to be accessible and respected as a symbol of global human heritage.

What was the significance of Hagia Sophia's conversion into a museum in 1934?

repositioned Hagia Sophia as a global cultural landmark.

- UNESCO and international leaders have raised concerns regarding ongoing preservation and interfaith accessibility at these historic monuments.