Abe International Symposium (4th Intl. Symp. on Oxidative Stress for Sustainable Development of Human Beings)

Many types of antioxidant supplements are available in the private market in Japan. However, it is difficult to know which type and how much to take, as it is possible to take too many of some vitamins. Since it is difficult for general consumers to make a choice, it is important to provide information based on scientific evidence. This study investigated the effects of continuous administration of a blended supplement, Twendee X (TwX) to aging mice. When 18-month-old C57BL/6 mice were given TwX for 1 month, behavioral tests showed that special cognition and short-term memory significantly improved compared to the age-matched controls. There were no significant differences in secreted neurotrophic factors, such as nerve growth factor and brain-derived neurotrophic factor in the brain. In treadmill durability tests before and after administration, the rate of increase in running distance after administration was significantly higher than that of the untreated group. These results suggest continuous intake of TwX may improve cognition and suppress age-related muscle decline. There is no problem with overdosing, so we think it's a good idea to take the blended supplement continuously.

Backgrounds: Oxidative stress is produced at the wound site, and excessive oxidative stress causes poor wound healing which can lead to dysfunction of the organ. The vocal fold is a vibratory mucosa in the voice box, and creates voice by high frequency vibration such as 100Hz to 800Hz. It can be injured by excessive voicing (vocal abuse), traume, and inflammation. It should be important to control oxidative stress during wound healing of the vocal fold to maintain the vibratory function keeping voice.

Materials and methods: Patients that underwent surgery to the vocal fold due to vocal fold lesions were enrolled in this study. They were seprated into 2 groups: a group treated by anti-oxidant, Twendee X, before and after the surgery (TWX group), and another group with no anti-oxidanta therapy (Control). Post-operative vocal functions were evaluated up to 3 months. The study was approved by institutional IRB.

Results: TWX group showed better vibratory properties with better phonatory function as compared to the control group.

Conclusion: TWX proved to be effective to improve wound healing of the vocal fold after surgery possibly due to reduction of post-operative oxidative stress.

Acne vulgaris is one of the most common skin diseases with varying etiologies. Acne vulgaris is most often seen on the face and back, and especially acne on the face is associated with a change in appearance, which may lead to psychogenic stress. Treatment includes cosmetic treatments such as chemical peels and phototherapy, but generally they are treated with topical medications, vitamins, or antibiotics for a certain period of time. Acne, including rough skin, is caused by oxidative stress due to fatigue, irregular lifestyle, and ultraviolet rays, which leads to the deterioration of the skin barrier function. SOD activity and malondialdehyde (MDA) concentrations in tissue and blood levels depend on the severity of the acne. Especially in severe acne, low SOD activity and high MDA levels have been reported, clearly indicating that oxidative stress is involved in acne vulgaris. Twendee X, an antioxidant supplement consisting of eight active ingredients of vitamins, amino acids, and CoQ10, has passed drug-level safety testing and is safe for use in children and adults. Since Twendee X significantly reduces blood oxidative stress in healthy individuals, it is feasible to provide oxidative stress care in acne vulgaris. In addition, each of the ingredients in Twendee X may have the potential to reduce the recurrence and aggravation of acne. We will report on the potential of Twendee X as an antioxidant treatment for acne vulgaris, using the results of a questionnaire we have conducted to date.

Free radicals continue to be produced in our body. Free radicals attack lipids and produce lipid hydroperoxide. Aggregated lipid peroxides are known to be a risk factor for developing various diseases such as arteriosclerosis and cancer. Antioxidant enzymes such as SOD and CAT that exist in the body are not sufficient to prevent these problems. Therefore, we consume antioxidants such as vitamins. Therefore, in this study, we reexamined the antioxidant activity of Twendee X, which is commonly sold as a multi-supplement [1]. In this experiment, electron spin resonance (ESR) was used to measure antioxidant activity. ESR is the only measurement method that can directly measure radicals. The results showed that Twendee X has very strong antioxidant activity. The ingredients contained in Twendee X are mainly water-soluble vitamins and amino acids. Despite this, it is surprising that Twendee X has the ability to scavenge hydroxyl radicals and superoxide radicals. This time, we will present a comparison of its antioxidant effect with other antioxidants. As future research progresses, we may be able to discover new combinations of vitamins and amino acids that strongly scavenge many types of radicals. Furthermore, if the relationship between individual radicals and disease and fatigue is clarified, it may become possible to create custom-made supplements.

This lecture will discuss the data showing that mammals, including the humans, have two major sources of melatonin that exhibit different functions. The best-known source of melatonin, herein referred to as Source #1, is the pineal gland. In this organ, melatonin production is circadian with maximal synthesis and release occurring during the daily dark period [1]. Of the total amount of melatonin produced in mammals, we speculate that less than 5% is produced by the pineal gland [2]. he regulation of the synthesis of pineal melatonin primarily involves the sympathetic innervation of the pinealocytes. Once synthesized, pineal melatonin is released into both the capillaries that perfuse the gland as well as into the cerebrospinal fluid (CSF) of the third ventricle, with both of these fluids exhibiting elevated levels of melatonin at night. The amplitude of the nocturnal rise in CSF melatonin is generally an order of magnitude greater than in the blood [3]. These melatonin rhythms have the primary function of influencing the circadian clock at the level of the suprachiasmatic nucleus (the CSF melatonin) and the clockwork in all peripheral organs (the blood melatonin) via receptor-mediated actions. A second source of melatonin (Source # 2) is produced in multiple tissues throughout the body, probably being synthesized in the mitochondria of these cells [4]. This constitutes the bulk of the melatonin produced in mammals and is concerned with metabolic regulation. Although this review emphasizes the action of melatonin from this source in determining redox homeostasis, it has other critical metabolic effects as well. The possible synthesis of melatonin in mitochondria is of particular interest since these organelles are a primary site of free radical generation [5].   Extrapineal melatonin synthesis does not exhibit a circadian rhythm and it is not released into the blood but acts locally in its cell of origin and possibly in a paracrine matter on adjacent cells [6]. The factors that control/influence melatonin synthesis in extrapineal cells have yet to be identified. We propose, however, that the concentration of melatonin in these cells is determined by the subcellular redox state and that it may be inducible under stressful conditions as is well documented in plant cells [7].

Dysphagia is a big issue for a large number of patients with cerebrovascular and other neurodegenerative diseases. Animal models are essential for understanding the pathophysiology of these conditions and developing effective treatments. In this study, we developed an animal model with attenuated pharyngeal constriction during swallowing using denervation of the pharyngeal branch of the vagus nerve. Our findings suggest that the pharyngeal area and pharyngeal transit duration during the pharyngeal stage of swallowing were increased compared to those in sham-operated and control animals. We investigated the potential of the anti-oxidant Twendee X in preventing oxidative stress caused by denervation-induced muscle damage, which could suppress muscle atrophy. Hence, we tested the effect of oral application of the Twendee X on swallowing function in the dysphagia model animals.

Our results indicate that Twendee X administration resulted in less increase in the pharyngeal area and pharyngeal transit duration compared to those in the dysphagia animal model. The thyropharyngeal muscles were also thicker than those in the nerve-sectioned animals. Overall, our findings suggest that Twendee X may have a possible role in preventing oxidative stress by the denervation of the pharyngeal constrictor muscle, leading to the suppression of denervation-induced muscle atrophy. Further studies are necessary to ascertain the clinical effects of Twendee X on bulbar paralysis in stroke patients. This study provides important insights into the potential use of Twendee X as a treatment for dysphagia patients.

Background: In recent years, fragrance pollution triggered by common household items has become a global concern, contributing to the increasing prevalence of Multiple Chemical Sensitivity (MCS) worldwide [1]. Despite the rising prevalence, there is a lack of researchers and diagnostic criteria for MCS, hindering effective diagnosis and treatment.

Purpose: This study proposes several investigations and experimental methods to elucidate the factors contributing to MCS and discusses the current status of diagnostic criteria for MCS, which remain unidentified.

Methods: We estimated the number of individuals affected by MCS based on existing studies [2] and identified fragrance ingredients commonly used in everyday products. Additionally, we explored methods to visualize invisible fragrances and considered how physicians should diagnose MCS.

Conclusions: At present, more than 16 million people in Japan (about 1 in 7) are estimated to suffer from “MCS” or have “High Sensitivity” or “Semi-High Sensitivity” to Chemical Substances. Recent developments in microencapsulation technology suggests that sustained fragrance release may contribute to the increase in MCS prevalence by continuously emitting hazardous substances [3], similar to allergic reactions seen in individuals with pollen allergies. Therefore, MCS should be recognized as a condition that anyone can develop, similar to pollen allergies. To prevent MCS, essential measures such as refraining from releasing fragrances in shared spaces are indispensable.

Background: Multiple Chemical Sensitivity (MCS) is a rising concern worldwide, particularly in Japan, where the number of individuals with high chemical sensitivity has increased by 500% over the past decade, with the current prevalence estimated to be 1 in 7 people. The exposure to fragrances in households continues to rise, as fragrance chemicals are found in nearly every household product. Limonene, an ingredient common to 77% of fragrance products, converts to formaldehyde in the air, which potentially implicates it in MCS pathology due to the generation of oxidative stress.[1][2]

Purpose: This study aims to investigate the relationship between a fragrance ingredient, formaldehyde generation, oxidative stress, and MCS pathology.

Methods: Over 40 Japanese detergents and fabric softeners were assessed for common ingredients, with limonene identified as the most prevalent. Gas detection methods were employed to measure the amount of formaldehyde generated from limonene.

Results: Heating limonene to 37°C produced formaldehyde concentrations exceeding indoor air quality standards, when the concentration of limonene was around 400 ppm (in the range of an easily detectable to strong odor). The concentration of formaldehyde surpasses permissible regulatory indoor standards and could increase oxidative stress in airway tissue and the blood.[3] This toxic effect potentially suggests a pathological mechanism for triggering MCS symptoms.

Conclusions: These findings highlight the potential role of common fragrance ingredients in formaldehyde generation in households. The formaldehyde concentration reached exceeded indoor safe standards, which presents a necessity to investigate the relationship with MCS pathology further, mediated by changes in oxidative stress levels in airway tissue and blood.[4]

Mitochondria, a powerplant of the cell, have developed an elaborate communication network within the cell communication with the nucleus and other subcellular organelles using a broad array of signaling molecules, including the components of the TCA cycle, reactive oxygen species, and other messenger molecules.  This mitocellular communication ensures an orchestrated response to everchanging energy demands and energetic stress, ultimately preserving cell survival. Using small molecules mild mitochondrial complex I inhibitors, we demonstrated that activation of mitocellular communication promotes health and lifespan in chronologically aged wild-type mice and in wild-type mice fed with a high fat diet, a model of accelerated aging. Efficacy of this approach was demonstrated based on increased survival, improved energy homeostasis in brain and periphery, reduced oxidative stress,  multiple behavior and cognitive tests, and biochemistry and systems biology approaches. These methods allowed to identify key mechanisms essential for health- and life-extending  therapeutics. Most importantly, such an approach results in the activation of multiple neuroprotective mechanisms mimicking a polypharmacy approach that is necessary to treat complex human conditions. Consistent with the hypothesis that improved aging will result in a delay of the onset of age-related neurodegenerative diseases, we demonstrated that treatment with mitochondria-targeted molecules blocked the ongoing neurodegeneration and cognitive dysfunction in multiple mouse models of Alzheimer’s Disease. Taken together, our data suggest that activation of mitocellular communication could be achieved with mild mitochondrial complex I inhibitors. This approach could be beneficial to promote health and longevity restoring mitochondria function and energy balance in brain and periphery [1-3].

Depression and other neuropsychiatric diseases are brain disorders that affect daily life. They are triggered by stress from changes in environment, relationships, finances, chronic illness, and other life obstacles. The number of patients with depression is on the rise worldwide, and in recent years, due to the COVID-19 pandemic, there is concern about a further increase. Oxidative stress (OS) plays an important role in depressive disorders in recent studies, including decreased serum antioxidant levels in depressed patients. Increased reactive oxygen species (ROS) and OS-induced dysfunction are associated with the etiology and progression of depression. However, at present, depression is commonly treated with many antipsychotic drugs, and few therapies have targeted oxidative stress. Twendee X®︎ (TwX), an antioxidant combination supplement with dementia-preventive effects for mild cognitive impairment (MCI) in Japanese, provides mitochondrial protection, maintains neurogenesis in the hippocampal dentate gyrus, increases brain autophagy and telomeres, in addition to antioxidant properties that cannot be achieved with a single ingredient. It is composed of eight vitamins, amino acids, and CoQ10, and has passed the same safety standards required of pharmaceuticals. In addition to lowering blood oxidative stress, TwX has been reported to improve quality of life, including defecation status and sleep quality, by acting on the gut microbiota. These various effects suggest that TwX is promising in terms of providing a new treatment option for neuropsychiatric disorders such as depression. Antioxidant therapy using effective antioxidant supplements is promising in terms of diversifying treatment methods in the treatment of depression, even in view of its different positioning from pharmaceuticals.

Reactive oxygen species (ROS) and free radicals work to maintain homeostasis in the body, and excessive ROS can damage the body's proteins, lipids, and DNA. Oxidative stress (OS) is the term commonly used to describe the imbalance between the generation of free radicals in the body and the ability of cells to counteract them. Accumulation of OS is aging, and OS also represents an important role for physiological homeostasis. Deviations from sustained redox signaling homeostasis are also now known to cause disease. The important relationship between OS and various diseases has been established, and OS is now at the forefront of research to elucidate pathogenesis. Despite this, so-called antioxidant therapy for diseases is still not widely used.

There are many types of ROS and free radicals, and each type has different properties. The widespread use of antioxidant therapy requires a level of antioxidants that can counter these. It is not clear whether OS induced the disease or was secondary to tissue damage derived from the onset of the disease. Although the exact role of oxidants in disease pathogenesis is not always clear, OS has received significant attention as a factor in human disease and is the focus of extensive research. This field will contribute to the prevention and treatment of diseases in the future.

After the onset of ischemia in an organ, treatments allow blood to flow back into the organs (reperfusion). Typical reperfusion procedures include thrombotherapy for cerebral infarction and catheterization for myocardial infarction. When blood begins to re-enter an ischemic organ, a large amount of oxidative stress is generated from the damaged area. In the case of cerebral infarction, prolonged oxidative stress causes inflammation of the surrounding normal cranial nerve tissue, leading to functional impairment and vascular dementia. In the case of myocardial infarction, it is known that even if catheterization allows blood to return to the heart, a large amount of oxidative stress is generated from the damaged myocardium, resulting in heart failure and death 5 to 7 days after the infarction.

There are limited methods of anti-oxidant treatment for reperfusion. Although Edaravone (RADICUT BAG I.V. Infusion) is covered by health insurance in Japan for cerebral infarction, it is only allowed to be administered once within 24 hours after the onset of cerebral infarction due to its strong side effects. Antioxidant therapy has been tested in myocardial infarction, but no significant effects have been reported.

We have developed an antioxidant combination supplemental, Twendee X (TwX), and TwX has been reported that it can reduce cerebral infarction damage in a mouse model of cerebral infarction. A small number of cerebral infarction patients have reported the improvements of sequelae and reduction of the severity symptoms at the time of infarction. In myocardial infarction, one patient with ST-segment elevation myocardial infarction who had been taking TwX before the onset of the disease was discharged from the hospital on the fifth day without symptoms of heart failure after catheterization. As a safe antioxidant therapy, TwX may be useful in reperfusion disease.

The vocal fold vibrates in high frequency to create voice sound. The vocal fold has a sophisticated histological “layered structure” that enables such vibration. As the vibration causes fricative damage to the mucosa, excessive voicing can cause inflammation or injury to the mucosa. Chronic inflammation or repeated injury to the vocal fold occasionally induces scar formation in the mucosa, which can result in severe dysphonia, which is difficult to treat. Oxidative stress has been proven to be an important factor in aggravating the injury, which can lead to scarring. It is important to avoid excessive oxidative stress during the wound healing period. Excessive accumulation of reactive oxygen species (ROS) has been found in the injured vocal folds of rats during the early phase of wound healing. Antioxidants proved to be useful in preventing the accumulation of ROS during the period with less scar formation in the long-term results. Oxidative stress is also revealed to contribute to aging of the vocal fold, in which the mucosa becomes thin and stiff with a reduction in vibratory capacity. The aged voice can be characterized as weak and breathy. It has been confirmed that ROS gradually increases in rat vocal fold mucosa with age, which may cause further damage to the vocal fold. Antioxidants have also proved effective in avoiding aging of the vocal fold in rat models. Recently, human trials have shown significant effects of the antioxidant Twendee X for maintaining the voice of professional opera singers. In conclusion, it is suggested that oxidative stress has a great impact on the damage or deterioration of the vocal folds, and the use of antioxidants is effective for preventing damage of the vocal fold and maintaining the voice.

Vocalization is a complex laryngeal function that involves intricate neuronal networks in the brain. This function depends on vocal fold vibration, which requires adequate subglottic pressure, vocal fold adduction, and tension. However, excessive use of vocal folds can damage the tissue structure of the vocal folds, as well as the laryngeal and respiratory muscles, possibly due to oxidative stress. Therefore, we conducted a study investigating whether vocal loading could lead to functional deterioration of the vocal-related muscles. 

Thus, we achieved an animal model, in which excessive vocal fold use induces hoarseness, produced by repetitive forced vocalization triggered by electrical stimulation of the midbrain periaqueductal grey in guinea pigs.

To examine oxidative stress of the laryngeal and respiratory muscles of vocal-loaded animals, we then compared the formation of malondialdehyde protein adducts of the laryngeal and respiratory muscles for a representative vocal-loaded animal with a control animal. The intralaryngeal and expiratory respiratory muscles showed higher levels of malondialdehyde in a vocal-loaded animal.

While additional experiments are required to substantiate this hypothesis, these results may give a new perspective on evaluating vocal fatigue in individuals who use their voices excessively. They may also help identify potential interventions or treatments for vocal disorders.

Airway reflexes such as coughing, sneezing, and the expiration reflex are essential in preventing foreign body from staying in the airway. These defensive reflexes should be appropriately activated against foreign bodies entering both the upper and lower airways. However, excessive responses to airway stimulation can lead to further airway distress and result in complications such as an overactive cough reflex and sneezing.

Allergic airway diseases, such as asthma and allergic rhinitis (AR), are typically chronic and are occasionally characterized by excessive and prolonged Th2 responses to inhaled allergens. They are assumed to be linked to oxidative stress. Asthma is associated with decreased antioxidant defenses, such as superoxide dismutase, catalase, and glutathione. Patients with AR have systemically elevated oxidative stress and systemically elevated serum total antioxidant status levels. Concomitant use of nasal steroids and antihistamines significantly decreases total oxidative stress in AR patients. Significant improvement in clinical outcome was observed in subjects who received antioxidants along with intranasal steroid fluticasone furoate. Other treatments that have been reported to improve symptoms of respiratory allergic diseases by enhancing antioxidant status include hydrogen-rich saline, crocin, curcumin, and silymarin. 

Interleukin (IL)-4 and IL-13 are critical cytokines in the induction of the pathogenic Th2 responses. They induce periostin in the airway tract that is highly expressed in chronic inflammatory diseases―asthma, atopic dermatitis, eosinophilc chronic sinusitis/chronic rhinosinusitis with nasal polyp, and allergic conjunctivitis. 

In this presentation, we will briefly review previous studies regarding airway disorders linked to oxidative stress. We will also introduce our recent project regarding airway hyperresponsiveness and the involvement of periostin in respiratory allergic diseases using periostin-knockout mice and respiratory allergic models. Further studies are necessary to evaluate the possibility of anti-oxidative treatment for the hypersensitivity caused by allergic airway inflammation.

Amyotrophic lateral sclerosis (ALS) is a disease that causes muscle weakness in the extremities, muscle atrophy, and dysphagia due to motor neuron degeneration. It is a neurologically intractable disease that mainly develops in middle age or later, eventually leading to respiratory failure due to paralysis of respiratory muscles, resulting in death within 3-5 years. Familial ALS is found in approximately 10% of all ALS cases and was reported to be caused by a point mutation in the gene for Cu/Zn SOD (SOD1), an antioxidant enzyme. It is hypothesized that oxidative stress damage caused by SOD1 abnormalities is deeply involved in the pathogenesis of ALS, and that oxidative stress may play an important role in the progression and worsening of the disease in ALS. Recently, the use of Edaravone, a radical scavenger, was approved for treatment in Japan the first time. Edaravone was developed as a treatment for acute cerebral infarction and is useful for cranial nerve and blood vessel protection by inhibiting inflammation in the brain. It also significantly improves motor and cognitive deficits for Alzheimer's disease in neurodegenerative disorders, reduces Aβ/p-Tau accumulation, and alleviates neuronal loss, oxidative stress, and neuroinflammation. Similarly, Twendee X, a known antioxidant supplement, also significantly improves motor and cognitive impairment and reduces Aβ/p-Tau accumulation by inhibiting oxidative stress in the brain, protecting mitochondria, and maintaining neurogenesis and autophagy function. It is composed of eight active ingredients consisting of vitamins, amino acids, and CoQ10, and has passed drug-level safety testing. Studies have shown that Twendee X has the potential for symptomatic relief of systemic scleroderma, an intractable therapeutic disease. Twendee X is not a pharmaceutical product and can be used safely on a daily basis. Twendee X is expected to be one of the most promising antioxidant therapies for ALS. Treatment experience for one patient of ALS also presented.

Fatty acid liver disease is a growing health problem associated with the increasing prevalence of obesity and diabetes. Elevated free fatty acid (FFA) concentrations are linked with the onset of peripheral and hepatic insulin resistance and, while their precise action in the liver remains unclear, it leads to liver steatosis. Although steatosis represents a reversible state of excess intra-hepatic lipid, it is also associated with increased susceptibility to oxidative stress and inflammation thought to trigger its progression to irreversible liver injury characterized by steatohepatitis, cirrhosis and hepato-carcinoma. The current molecular mechanisms of this progression remain poorly understood. However, the “two hit” hypothesis represents the most commonly accepted model. In this model, steatosis represents the first “hit”, sensitizing cells to subsequent stress with a dysregulation of energy production and accumulation of ROS. The second “hit” may take many forms, including drugs, hypoxia or cytokines, eventually leading to inflammation or steatohepatitis. Few in vitro models exist that can recapitulate this progression and its dynamics. Twendee X (TwX) is a potent anti-oxidant and that it is capable of reducing H2O2-induced and acetaldehyde-induced oxidative stress in native HepG2 cells. We have established the FFA-induced lipid accumulation in HepG2 cells, as well as ascertained that the model induced significant oxidative stress and perturbed mitochondrial bioenergetics. We also established the effect of TwX treatment in dose response in both preventive- or curative-treatment designs, and further obtained evidence of intracellular signaling pathways involved both in the FFA-induced oxidative stress and in TwX activity in regulating and normalizing these pathways.

Sinusitis is a disease that is accompanied by a runny or stuffy nose, pain in the cheeks, between the eyes, and in the head, and olfactory disturbances These symptoms can lead to a decreased quality of life due to lack of concentration and discomfort. The main cause is viral or bacterial infections, such as the common cold. Since the sinuses are connected to the nasal cavity, it is known that infection can also cause inflammation of the sinuses. When the body is invaded by a virus or bacteria, it employs highly active reactive oxygen species (ROS) to eliminate them. This causes inflammation. Therefore, ROS are elevated in areas of inflammation, resulting in increased oxidative stress. In patients with chronic sinusitis, reduced glutathione and uric acid concentrations have been reported, suggesting that chronic sinusitis is likely to play a role in oxidative stress. Twendee X is an antioxidant supplement that contains a balanced blend of eight ingredients with strong antioxidant potential. It has passed drug-level safety testing and is safe for use by both children and adults. This study reviewed the relevance of oxidative stress in sinusitis and the results of a questionnaire on symptom changes in humans with sinusitis before and after taking Twendee X. The results suggest that intervention with antioxidant supplements could improve or prevent symptoms of sinusitis.

The telomere length is suggested and used as a biomarker of human aging simply due to previously telomeres has been suggested to predict longevity. Oxidative stress is presumably one of the major causes of telomere shortening, 

Our findings supported the idea of a possible correlation between the TL and biomarkers of oxidative stress in aging. The study has remarkable scope in medical science as the findings on correlation of TL with biomarkers of oxidative stress in aging are novel and they will help in further research against oxidative stress.

During aging, telomeres shorten classically due to cell turnover. Telomere length is mainly maintained by telomerase. This enzyme is present in the embryonic stem cells in high concentrations and declines with age. It is still unclear to what extent there is telomerase in adult stem cells, but considering these are the founder cells to the cells of all tissues in a body, understanding the telomere dynamics and expression of telomerase in adult stem cells is very important.

Telomere length has been implicated as one of the markers for aging related diseases and neoplastic transformation in both in vivo and in vitro studies. During carcinogenesis telomeres shorten due to high cell turnover and repeats are added by active telomerase or alternative lengthening of telomeres (ALT).  This gradual shortening is replication driven and does not necessarily explain the presence of ultra-short telomeres. 

Ultra-short telomeres are observed when there is a sudden shortening in telomeres not related with cell division and may arise from breaks in telomeres due to oxidative damage and replication slippage. Telomeres have important functions but do shorten through-out life, ultimately causing cellular problems. 

Our group has compared different methods that available to evaluate telomere length, with a special focus on the telomere length dynamics in different tissues, both the overall telomere length and telomere length of individual chromosomes in age related disorders. 

Thus, our results showed that telomere profiling may be use as an important clinical parameter and supported the idea of a possible correlation between the ultra-short telomeres as biomarkers of aging. Overall telomere science showed that single or a small group of ultra-short telomeres are more influential in senescence associated disease progression rather than shortening that reflected as average telomere length, therefore it is important to identify the presence and load of ultra-short telomeres in diseases.

Our results suggest the using Universal STELA is an accurate method for evaluation of extreme-short telomeres. Compared to golden standard well known TRF assay, that measures mean telomere length, U-STELA is developed to overcome several problems detecting abrupt telomere shortening in a single chromosome out of 92 chromosome ends same time. The novel approach in U-STELA is to anneal a linker or telorette to the G rich 3’ overhang of the telomere which is a product of restriction digestion after DNA isolation. Telorette enables stable PCR of telomeric regions without template slippage ensuring successful completion of PCR.

A collection of numerous casses and stories worldwide will be described on the effect of Twendee X and MTControl on the people’s life. They range from improving the quality of life of normal healthy people at various stages of their life and more importantly on people that suffer from various diseases before, during or after treatments. This collection help develop scientific placebo based studies on the effect of Twendee. The success stories are numerous and impressive.