Despite extensive studies, cancer remains one of the most dreadful diagnoses and biggest challenges for human health all over the world, representing a leading cause of death in the industrialized countries. Various chemotherapeutic drugs, such as doxorubicin, tamoxifen and paclitaxel, have been used for the treatment of tumors for more than half a century; however, there are still no curative options currently available in clinical settings and the severe adverse effects of these drugs threaten the well-being of the patients seriously. Current evidence suggests that further knowledge is urgently needed to clarify the unknown properties and molecular mechanisms of action of various chemopreventive molecules. Such substances refer to the agents which are used for reducing the risk of carcinogenesis, or delaying development or recurrences of malignant disorders.in silico tools, such as docking and QSAR (Quantitative structure-activity relationship) techniques, can be used to retrieve more comprehensive information about the targets and the action mechanisms of such molecules. However, there has been a vigorous need to explore the acute as well as the chronic toxicological effects of such chemopreventive molecules for further clinical implementation against diseases.Several small molecules either of biological origin or from synthetic chemistry, such as cordycepin, mitomycin C, doxorubicin and methotrexate, have demonstrated great efficacy towards a variety of cancers. These agents are found to suppress the proliferation of cancer cells by various mechanisms such as apoptotic cell death, cancer cell cycle arrest, inhibition of angiogenesis and metastasis, induction of ROS generation etc. Few Singh et al. about potential therapeutic significance of 4-(methylthio) butyl isothiocyanate (4-MTBITC) on modulation of glycolytic enzymes and hypoxia pathway in female rats. As increased glycolysis is known to be an indicator of malignancies  shows multi-pronged therapeutic potential of this extract against HPV infection and cervical cancer. Infection with high-risk HPV subtypes is generally accepted as a risk factor for cervical carcinogenesis . In a further article, Baba et al. review the dual role of TGF-β under different cellular conditions and its crosstalk with other signaling pathways in modulating cell fate. As a pleiotropic cytokine, TGF-β can act both as a tumor suppressor as well as tumor promoter depending on the context and stage of tumor progression , quercetin and grape seed procyanidins, among several other bioactive natural compounds, in the models of different cancer types. Hu et al. describe the contribution of lncRNAs and miRNAs in triple-negative breast cancer pathogenesis, proliferation, migration or malignancy. Finally, Rah et al. review the role of JAK/STAT pathway to improve the existing cancer therapies. Modulation of this signaling cascade might be important also in cancer prevention. We hope that you enjoy the reading of this article collection, composed in a topic that will probably be even more important in the coming years.This Research Topic aims to highlight the ongoing advancement in chemopreventive and therapeutical approaches, as well as the promising role of the above-mentioned agents in the context of cancer prevention and therapy. In particular, it encompasses the research of gnancies , inhibitogenesis , highliggression . Fined-t


As the world population is aging, intervertebral disc degeneration (IDD) is becoming a global health issue of increasing concern. A variety of disc degeneration diseases (DDDs) have been proven to be associated with IDD, and these illnesses have significant adverse effects on both individuals and society. The application of stem cells in regenerative medicine, such as blood and circulation, has been demonstrated by numerous studies. Similarly, stem cells have made exciting progress in the treatment of IDD. However, due to complex anatomical structures and functional requirements, traditional stem cell injection makes it difficult to meet people’s expectations. With the continuous development of tissue engineering and biomaterials, stem cell combined with biomaterials has far more prospects than before. This review aims to objectively and comprehensively summarize the development of stem cells combined with contemporary biomaterials and the difficulties that need to be overcome. The intervertebral disc (IVD) is the cartilaginous tissue that lies between the spinal segments and plays a crucial role in maintaining the normal physiological activity of the spine. With the increase in aging populations worldwide, intervertebral disc degeneration (IDD) is becoming a health problem that is difficult to ignore or compromise. IDD could lead to disc degeneration diseases (DDDs), such as compression of blood vessels or nerves, and disc herniation . DDDs ofThe specific causes of disc degeneration are not fully understood. The prevailing view is that IDD is a combination of factors, such as age, genetics, and diabetes . TypicalMany promising therapeutic approaches have been developed for the various mechanisms of the IDD, such as gene therapy, biological agents, and growth factors. Some of them have been in pre-clinical trials while others have been approved for clinical use . HoweverIVD is the largest avascular tissue in the human body, which is mainly composed of nucleus pulposus (NP), annulus fibrosus (AF) and cartilage endplate (CEP) . Due to 6 cells/ml, and it further decreases with age , poly (ethylene glycol), and poly (lactic-co-glycolic acid) copolymers. These materials can be industrially produced in large quantities and have strong mechanical properties, but their cytotoxicity and immunogenicity should be considered. Composite hydrogels can combine components with different properties to achieve more ideal performance. However, the involvement of different components bring new problems to the overall material design. The Several decellularized scaffolds have been developed. To better mimic the component composition of normal disc tissue. However, the scarcity of healthy donor sources that are suitable for humans limits their further development, and the use of animal-derived substitutes requires strict safety assessment. The microsphere system can control the release of substances in the delivery system in a more precise way, in addition to providing attachment sites for cells. Some of the common microsphere materials include natural materials such as gelatin, collagen and chitosan, as well as polylactic acid-glycolic acid, poly (L-lactic acid) and polycaprolactone. Unfortunately, it is difficult for the microspheres to provide mechanical support so other materials should be introduced, though this makes system design more difficult.In recent years, some artificial polymer materials significantly improved the performance of biomaterials, with a remarkable balance in mechanical strength and porosity. Increased understanding of the structure of IVD and continuous improvement of the material preparation technology has made it possible to imitate the structure of tissues to provide a more suitable environment for stem cells. Additionally, scaffolds that are loaded with drugs or cytokines are better applicable cell differentiation and expression of corresponding products.Most tissues and cells in the human body are undergoing constant renewal, and sufficient cells plays a crucial role in maintaining normal tissue function and metabolism. This rule also holds true for the IVD tissue. Again, it is worth noting that the low cell density, lack of a powerful source of regenerative cells, and the harsh microenvironment after IDD make the endogenous self-cell recruitment of IVD difficult. In degenerated IVD, cell loss was reflected by higher rates of senescence, apoptosis, and pyroptosis. Direct replacement of the missing cells seems to be the most straightforward and effective method, though there are still many limitations, such as the survival rate of the implanted cells and unnecessary cell dissipation. Stem cells combined with biomaterials offer new hope for stem cell-based endogenous repair.et al.  Earlier, faster, more economical screening techniques for IDD, as well as comprehensive and easy-to-accept patient education.Future research may focus on the following aspects: 1) A deeper understanding of the pathological mechanism of IDD and more effective therapeutic targets will provide more succinct directions for selecting drugs and preparing materials; 2) Further exploration of stem cell repair mechanisms, as well as efficient and stable differentiation induction methods; 3) Identification of materials with good properties that can withstand different mechanical requirements in various areas of IVD, in addition to providing a suitable microenvironment for cells; 4) Development of IDD models that closely imitate human IVD environment development and the corresponding


The aim of this report is to present a case of a patient who developed unusual systemic hypersensitivity reaction to a red-pigmented tattoo and to discuss diagnostic difficulties in case of systemic reactions to tattoo ink. The patient reported erythroderma on his arms and chest accompanied by plaque elevation of red parts of his most recently performed forearm tattoo as his primary symptoms. His health condition entailed a prolonged topical and intravenous immunosuppressive therapy, which proved ineffective. Over a year after emergence of initial symptoms, he presented to the Plastic Surgery Clinic with generalized erythroderma, systemic lymphadenopathy, elevation and granuloma formation in red tattoos on his forearm and complaints of fatigue and inability to participate fully in work-related and social activities. The patient underwent six staged excisions with direct closures, flap plasties and full-thickness skin grafts. Following completion of each surgical resection, the patient’s symptoms gradually subsided. We find this case illustrative of a clinical challenge that delayed hypersensitivity reactions to red tattoos may pose. Furthermore, we provide insights on management of hypersensitivity reactions. This report underlines the importance of social awareness of and public health approach to tattoo complications as key to successful prevention, identification and treatment of adverse reactions to tattoos. Tattooing is a millennia-old form of body modification performed by depositing pigment in the dermis layer of the skin. It may be perceived simply as a decorative marking on the body or serve as a way to enhance one’s sense of individuality, increase attractiveness or express affiliation to a subculture . The numThe aim of this report is to present a case of a patient who developed unusual systemic hypersensitivity reaction to a red-pigmented tattoo and to discuss diagnostic difficulties in case of systemic reactions to tattoo ink.A 36-year-old Caucasian male was referred to the Plastic Surgery Clinic by an allergist. He presented with skin changes  on red parts of tattoos on his right forearm and complained of fatigue and reduced quality of life. The patient had many extensive colorful tattoos on his chest and both upper limbs, which were performed a few years earlier, except for red, black and green tattoos on his right forearm, which were performed about 6 weeks prior to the emergence of systemic erythroderma and cutaneous changes . He had The patient’s medical history includes Hashimoto’s thyroiditis treated with levothyroxine, a period of self-administered testosterone misuse for muscle-mass building purposes, and hospitalization for pneumonia associated with COVID-19 infection during immunosuppressive treatment of erythroderma and skin changes. His family history includes hypothyroidism. He has no known history or family history of autoimmune conditions, such as sarcoidosis or dermatologic diseases.Bloodwork revealed mildly elevated C-reactive protein (CRP) (8.9 mg/L) and eosinophilia . Diagnostic tests that the patient underwent also included biopsy of an enlarged lymph node from his groin, which showed deposits of red pigment and lymphocytic infiltration. He also underwent multiple skin biopsies of the reactive tattoo area and nontattooed skin affected by pruritis at various stages of treatment. Nontattooed skin specimens yielded nonspecific findings, while the tattoo bioptates showed pigment deposits with intensive inflammatory cell infiltration and erosion throughout the epidermis and skin appendages. Biopsies did not show signs of lymphoproliferative diseases. He tested negatively for HIV, HBV, HCV, parasites, and did not have any remarkable findings in computer tomography imaging of the chest, abdomen and pelvis. Patch testing for metal allergy yielded negative results.Until the onset of skin lesions, the patient was a semiprofessional athlete, maintained good health condition and was subject to regular medical supervision. Medical interview revealed that the first symptom—erythroderma—appeared slightly over 1.5 months after tattoos on his left forearm were made and coincided with the time when the patient was delegated to work in a COVID-19 hospital as a professional soldier assisting medical staff. It was at the time of the first wave of the SARS-CoV-2 pandemic, before vaccinations against the disease were available. The position required him to wear protective clothing and expose his skin to substantial amounts of disinfectants, which had an irritating effect and caused outbreaks on the skin. Initial erythema on the forearms and torso progressed into painful erythematous, populous and squamous lesions on his head, neck, torso and limbs with pruritis, and hyperkeratosis of the soles of the feet and hands. Associating skin changes with the nature of his work prompted the patient to undertake testing for COVID-19, which yielded negative results. The patient resigned from work at the COVID-19 hospital to limit his exposure to irritants, yet the symptoms did not resolve. Over the course of the year, he looked for treatment at dermatology, internal medicine, hematology, and allergology departments to relieve his symptoms. Given the untypical, challenging clinical presentation and emergence during the COVID-19 pandemic, the diagnosis of chronic allergic tattoo reaction was not an easy one to reach, and, thus, multiple diagnoses were suspected, including atopic dermatitis, pityriasis rubra pilaris, follicular mucinosis and lymphoma.The patient underwent several courses of conservative treatment, which included oral and intravenous immunosuppressive therapy , UVA1 phototherapy, retinoids, antibiotics, protective and lubricating skin ointments, and oral antihistamine drugs. The symptoms partially improved on high doses of intravenous immunosuppressants but recurred with each attempt to reduce the dose. After 10 months of treatment with only minimal effects, an allergist managing the patient grew suspicious of the appearance of the tattooed skin on the patient’s right forearm and considered his symptoms might be attributed to hypersensitivity reaction to red pigment in the ink used to perform one of the tattoos. Due to his working diagnosis, he referred the patient to our Plastic Surgery Clinic, where, after examination of the patient and his medical history, the patient was scheduled for surgical excision of the tattooed skin of his left forearm . Steroid3/μL, 13%).The result of surgical intervention was a complete excision of all reactive parts of the tattoos within the course of six surgeries performed in approximately one-month intervals. The resection was accomplished with the use of direct closures with presuturing, local flap plasties and full-thickness skin grafts harvested from the patient’s groin area. After each excision session, the healing period was uneventful and, with each resection performed, the patient’s systemic symptoms improved, with a significant relief of the symptoms following final resection. At a follow-up visit, three months after completion of surgical treatment, the patient’s recovery had been progressing with no recurrence of erythroderma . MoreoveTattoos, as any form of introducing a foreign body into the skin, might cause adverse reactions. The response might be of diverse nature and, aside from fairly easy to manage acute complications, such as photosensitivity or infection, common and problematic in their nature are chronic tattoo allergic reactions. As in the case of our patient, the major culprit of delayed allergic response to tattoo is red ink. Historically, red pigment was largely produced with the use of metals, including highly toxic mercury derivatives. These have now been replaced by less dangerous cadmium and ferric compounds, as well as organic substances  . DespiteTreatment in allergic tattoo reactions typically starts with topical, oral or intralesional corticosteroids and antihistamines. Sometimes laser removal is taken into consideration. These methods, however, are often ineffective in treatment of inflamed, thickened dermis in pigment-laden, reactive tattoos . In partTattoo ink is a frequent cause of delayed hypersensitivity reaction and potential serious sequelae should be taken into consideration when deciding on getting a tattoo, particularly made with red ink. Our case is illustrative of serious and untypical complications red tattoos may give rise to. Care should be taken upon examination of systemic skin changes in tattooed patients, as failure to reach a proper diagnosis puts them at risk of prolonged therapy and its side effects, and may significantly impair their quality of life. Upon detection of a chronic allergic tattoo reaction, surgical excision should be taken into consideration as an effective symptom-relieving treatment. Additionally, the report underlines the importance of social and clinical awareness of tattoo complications, especially in the era of rising popularity of tattooing.


Corporate capture is the deliberately planned process whereby political decisions respond to a particular interest of a private nature, in detriment to the public interest. The result is an unjust regulation or the absence of regulation where this is necessary for the protection of the common good. Corporate capture of public health has to do with commercial determinants of health, such as alcohol, tobacco, sugar-sweetened beverages and ultra-processed foods. Capture-related actions are targeted at civil society, experts, public-health officials, civil servants and politicians. These actions range from material  to intellectual , social (control of information and communication) and/or cultural . The most common capture strategies are aimed at biasing scientific results, creating consumers from an early age, promoting a good image of corporations, questioning the legitimacy and appropriateness of governmental intervention aimed at regulating their activity, controlling professional education; and exerting pressure on governments and international bodies. To illustrate this phenomenon, we present findings from several examples of corporate capture of food policies in Europe. Preliminary results from publicly available information suggest that most of the mentioned tactics were used in Europe in order to block nutritional profiles, health warnings on food and effective food advertising regulations. We suggest implications for how European legislation can better protect European citizens, especially children, from vested interests aimed at promoting the consumption of unhealthy food and beverages.


Dental implants and related bone augmentation problems have seen major progress since early protocols were tested in the 1980s. However, major challenges still exist concerning hard tissue augmentation procedures. Indeed, more than half of the patients show bone defects and require augmentation solutions for implant installation.The bone augmentation procedures experience healing and maturation complexity because of the biology and physiology of soft and hard tissues, the patients’ medical risk factors, the grafting materials employed and the complexity of surgical techniques used to implant them .Therefore, conventional bone grafting materials can bring disadvantages, because of the brittleness of allografts, xenografts, and alloplasts, uneasy porous forms generation, and the inability to generate precise patient-specific structures to comply with the need for precision medicine. In the meantime, autografts cannot be easily shaped to meet bony defect demand .Additionally, the acceptance of bone augmentation materials can be affected by religious or life ethics—vegetarian or vegan—personal choices, as well as allergies, and refused when animal-derived products are proposed.The challenges for researchers are, therefore, to explore novel bone graft substitutes to be used as allografts, xenografts, and alloplasts, or therapy able to prevent bone resorption and/or support bone augmentation.Certainly, new biomaterials and their employment in the printing of 3D scaffolds are one the more interesting support structures for the future of implant dentistry.Biodegradable natural polymers in association with bioactive and/or mechanically strong materials showed the potential to combine bioactivity and modelling. The application of 3D-printed scaffolds in tissue regeneration, and particularly in bone and sinus augmentation, and socket preservation suggested promising outcomes [Behind or alongside 3D techniques, novel therapies from collaborative physical, biomolecular and pharmaceutical research could support and improve bone augmentation medical requests.Photobiomodulation has recently demonstrated its reliability in the field of regenerative medicine. The ability of light in the visible and near-infrared spectrum to stimulate endogenous cell targets and modulate its energetic metabolism influencing the stem cell and osteo-precursor agenda was recently reviewed . PromisiLastly, recent research points out the interest in dopamine behavior as an immunomodulatory mediator of several immune cells and its possible target role for drug therapy acting on osteo-inflammation and bone degeneration. Particularly, treatment with a dopamine-2-type receptor agonist is able to counteract inflammation and stimulate bone regeneration and support the transplantation of stem cell bone precursor efficiency .Collectively, the recent background on bone augmentation for implant dentistry reinforces the need to develop meaningful clinical endpoints supported through the collaboration of a wide field of scientific research in biology, pharmacology, physics, chemistry and engineering. The key role of translational medicine will be mandatory in light of compliance with precision medicine and supporting the needs of clinicians, private-practitioners, and patients.


Multiple myeloma (MM) is a prevalent hematological malignancy. Resource-constrained settings such as the Middle East are particularly burdened by the increasing trends in MM morbidity and mortality in addition to challenges in the management of MM. It thus becomes necessary to identify and address debatable areas of current practice and gaps in the management of MM in the Middle East. With a special focus on the Lebanese situation, the first-line treatment of the very elderly (> 80 years old) is discussed, in addition to the impact of relapse type  on maintenance therapy, the choice of first relapse therapy in relation to maintenance therapy, and the role of MRD in the MM treatment landscape. The need for realistic management guidelines accounting for local resources and expertise, in addition to the reflection of drug accessibility and cost on clinical practice are recognized. Multiple myeloma (MM) is one of the most prevalent hematological malignancies, with 176,404 new cases and 117,077 deaths reported globally in 2020 . MM trea. Dose reduction of novel drugs and schedule adjustment can be beneficial with similar outcomes in unfit patients. Furthermore, in the modern era, several studies have demonstrated the toxicity of long-term use of steroids in non-transplant eligible patients and emphasized the value of decreasing the dose and/or reducing the duration of steroid administration with lower toxicity and mortality [. With the risks of melphalan-based regimens in the very elderly, novel drugs should be considered, if available, either as part of treatment or maintenance therapy. This does not preclude the consideration of older drug regimens, which are also effective as maintenance for elderly MM patients [Current real-world evidence shows that triplet therapy followed by autologous hematopoietic stem cell transplantation, consolidation and maintenance for fit responder patients remains the backbone of MM treatment in Lebanon, while novel regimens are being explored whenever possible . Fit patortality –16. Withpatients . Front-lpatients . HoweverThe 2021 European Hematology Association (EHA)-European Society for Medical Oncology (ESMO) and the IMWG guidelines offered detailed recommendations for the management of relapsed MM patients , 20. In However, even with the availability of treatment recommendations, the question remains: which drugs are available and when should relapse therapy be initiated? Drug availability is an important factor and treatment guidelines such as the 2021 IMWG recommendations are progressively making provisions for drug access limitations in low- and middle-income countries . The 201Younger, fit patients who are eligible for transplant should be considered for ASCT, seeing as it remains one of the most effective and widely available therapies for MM. Tandem ASCT as consolidation can also be considered only for high-risk cytogenetic patients . MaintenMRD negativity has notable prognostic implications , and shoWhen treating MM, patient-related and treatment/disease-related risk factors should be carefully considered, and the management of patients should be individualized accordingly. Gaps in oncologist knowledge and education should be addressed to aid in the personalization of treatment and the integration of evidence-based recommendations into real-world practice. Official, clear and updated guidelines are needed to guide the management of MM patients in Lebanon. Treatment initiation in patients with biochemical relapse without end-organ involvement might be beneficial but remains debatable. The combination of novel agents with standard of care is promising for the induction, consolidation, and maintenance of MM patients. Nevertheless, accessibility and cost should be carefully considered, and realistic management guidelines are needed to optimize diagnosis and treatment efficacy according to local resources. Very elderly patients should not be denied treatment, and adjusted intensity regimens can be used in case of frailty. Achieving MRD negativity is an important treatment goal and carries significant prognostic value. The potential role of MRD-negative status as an outcome measure influencing treatment decisions in local populations, and the feasibility and cost-effectiveness of MRD assessment in everyday practice remains to be explored in the Middle East.


Cutting-edge technologies such as additive manufacturing (AM) have had an enormous impact in a multitude of sectors. The production of drug delivery systems and precise medicine are taken to another level with the application of AM in the pharmaceutical field. Using these super-trend production methodologies, it is possible to quickly produce personalized drug delivery systems that would be impossible to achieve using conventional manufacturing methodologies.Therefore, the reviews presented in this Special Issue inform readers of the different studies performed in recent years concerning the application of different printing methodologies for drug form production as well as different ink development. As one of the reviews published in this Special Issue, Dr. Billa and Dr. Khalid focus their manuscript on current trends in the application of AM to prepare personalized dosage forms on-demand, concentrating on coupling solid dispersion with fused deposition modelling 3D printing . On the This Special Issue was also furnished with interesting and innovative research papers that could lead to advances in the field of pharmaceutical/drug delivery development and production. A prime example is the study presented by Jung et al. (2021), where high-resolution and high-dimensional microneedles were successfully produced by adjusting the printing angle using a general SLA 3D printer. This technology could be applied to the manufacture of drug delivery tools and various microstructures .Additionally, an array of materials could be conjugated and optimized to accomplish improved drug delivery strategies for different kinds of applications, and these materials/structures must be well-characterized. In this regard, Tesk et al. conducted analyses of the biological, chemical, mechanical and thermal properties of photopolymerized poly(ethylene glycol) diacrylate (PEGDA) and specific copolymers with different photoinitiator concentrations before and after applying a post-treatment washing process. The main goal of this study was to provide readers with information concerning PEGDA multi-materials to contribute to the future development of tools ensuring safe and effective individual therapy for patients . To evalConcerning the production of different drug delivery structures using AM, this Special Issue presents an array of research papers focusing on different kinds of structures, different AM production methodologies, and different applications/targets. In this regard, fibres and nanofibres for different purposes were developed in both cases using electrospinning. In the first case, nitric-oxide polymeric fibres were developed to promote an antimicrobial effect , and eleAnother approach to a drug delivery structure could be a scaffold, which was the chosen methodology of Sandri et al. In this study, a medicated osteoinductive and bioresorbable bone graft was designed and investigated for its ability to control antibiotic drug release in situ. This represents an ideal solution to the eradication or prevention of infection, while simultaneously repairing bone defects. Overall, the produced scaffolds were safe and effective as a local delivery systems for an extended duration of therapy. Additionally, promising results for the prevention of bone-defect-related infections in orthopaedic surgeries were reported . GelinskResearch concerning the production of oral forms using AM is also presented in this Special Issue. One study focuses on the production of buccal films made of two synthetic polymers, gelatin-polyvinylpyrrolidone and gelatin-poly, and their characterization. The in silico population simulations indicated increased drug bioavailability and decreased inter-individual variations in the resulting pharmacokinetic profiles compared to immediate-release tablets . Saleh eCounterfeit products in pharmaceutical production is a major subject that is causing great concern. Therefore, a study developed by Quodbach et al. (2021) is presented, where a proposal to solve this problem using AM is made. To test this concept, the authors encoded binary digits (bits) on the surface of fused deposition modelling (FDM) 3D-printed geometries. The most commonly used polymers in pharma were tested, and the scanning and printing processes were evaluated. The main conclusions were that the addition of a colourant or active pharmaceutical ingredient could facilitate this detection process, and the process could also be transferred to 3D-printed pharmaceuticals. However, further improvements are necessary to increase robustness and allow the use of more materials .Moreover, the application of mathematical models in the development of drug delivery systems is also an advantageous approach since, in most cases, it is possible to decrease the costs involved in the drug delivery development process. As an example, Sousa et al. (2021) applied design of experiments (DoE) to quickly and less expensively explore and tailor the characteristics of chitosan/pDNA nanosystems to produce a DNA vaccine . ConcernOverall, the reports presented in this Special Issue highlight the importance of exploring production approaches based on the use of AM and mathematical models to achieve efficient materials, structures and devices to be applied in the drug delivery field. We, as Guest Editors, are deeply grateful to all the authors for their outstanding-quality research and the critical evaluations of their manuscripts.


Plants evolved an impressive arsenal of multifunctional specialized metabolites to cope with the novel environmental pressures imposed by the terrestrial habitat when moving from water. Here we examine the multifarious roles of flavonoids in plant terrestrialization. We reason on the environmental drivers, other than the increase in UV-B radiation, that were mostly responsible for the rise of flavonoid metabolism and how flavonoids helped plants in land conquest. We are reasonably based on a nutrient-deficiency hypothesis for the replacement of mycosporine-like amino acids, typical of streptophytic algae, with the flavonoid metabolism during the water-to-land transition. We suggest that flavonoids modulated auxin transport and signaling and promoted the symbiosis between plants and fungi , a central event for the conquest of land by plants. AM improved the ability of early plants to take up nutrients and water from highly impoverished soils. We offer evidence that flavonoids equipped early land plants with highly versatile “defense compounds”, essential for the new set of abiotic and biotic stressors imposed by the terrestrial environment. We conclude that flavonoids have been multifunctional since the appearance of plants on land, not only acting as UV filters but especially improving both nutrient acquisition and biotic stress defense. Many evolutionary innovations, i.e., the adaptive traits of living organisms , occurreFor instance, the two- (2D) to three-dimensional (3D) growth transition was a key innovation increasing the plant complexity ,12 and hThe outstanding ability of terrestrial plants to adapt to this “ever-changing” new environment has depended not only on the evolution of a pre-existing molecular toolkit but largely on the impressive chemical diversity originated by the rise of a huge number of specialized metabolic pathways ,33,34. TIn this brief review, we examine the multi-functionality of flavonoids, the ancient class of secondary metabolites synthesized through a branch-pathway of the general phenylpropanoid metabolism, which is unique to land plants ,42, in tOne of the most intriguing molecular innovations that accompanied the water-to-land transition of plants was the replacement of the mycosporine-like amino acids (MAAs) metabolism, typical of streptophytic algae, by the flavonoid biosynthetic pathway , a commoThe replacement of nitrogen-containing MAAs with carbon-based products synthesized by the “late” flavonoid pathway, particularly the flavonol branch, was, therefore, a molecular event of outstanding significance during plant terrestrialization ,48,64, aAs outlined above, one of the most severe challenges faced by early land plants was the drastic reduction in the availability of water and nutrients ,66. OnceAuxin is a major regulator of plant growth and developmental processes, and it is a central modulator/regulator in AM symbiosis  Figure a. IncreaThe strong dependence of AM on the auxin perception and signaling highlights the importance of endogenous regulators of this hormone, such as the flavonoids ,95. Actu2O2 and a range of protein kinases, including PID and mitogen-activated protein kinases (MAPKs) [Overall, this is consistent with the relatively old suggestions that flavonoids play a crucial role as developmental regulators ,110, whi (MAPKs) ,110. In Marchantia polymorpha and of quercetin derivatives present in Physcomitrium (previously Physcomitrella) [Our analysis leads us to hypothesize that not the increased UV-B irradiance, but the nutrient and water scarcities were the primary environmental drivers for the rise of flavonoid metabolism in substitution of MAAs during plant terrestrialization . This viitrella) ,115,116.itrella) , thus maitrella) ,117, mositrella) , and in itrella) ,119,120)M. polymorpha [P. patens [HY5 gene, is conserved in liverworts, mosses, and angiosperms [HY5 enhances the expression of transcriptional regulators of flavonol biosynthetic genes  [HY5 and its homolog HYH regulate not only the UV-B responses but also those triggered by blue and red light through flavonoid accumulation [As outlined in the previous sections, early plants were challenged against the concomitant action of multiple “new” environmental pressures when moving from water to land, high UV-B radiation being just one of these stress agents . The inelymorpha , and the. patens  also offiosperms ,124. Remd MYB13) ,126,127,d MYB13) ,120. Mormulation  reinforcThe location of flavonoids in mesophyll cells, in their chloroplasts, and even in the nuclei ,120 alsoWhen moving on land, plants were also challenged against a new set of biotic stress agents ,136. IndWe conclude that the replacement of MAAs with flavonoids during water-to-land transition equipped basal land plants with compounds capable of serving multifarious roles in response to the novel abiotic and biotic challenges imposed by the terrestrial habitat. Although the ability to perform more than one role is a common feature of most secondary/specialized metabolites, which follows nature’s strategy to catch as many flights with one clamp as possible, this is particularly relevant for flavonoids. Even though the evolution of flavonoid metabolism has probably expanded its functional roles in plant-environment interactions, we infer flavonoids have probably been multifunctional since the first appearance of plants on land.Plants were challenged against a wide range of novel environmental pressures when moving to land. The excellent capacity of plants to adapt to the harsh terrestrial habitat has depended on both the evolution of a pre-existing molecular toolkit and the rise of a huge number of specialized metabolites. Among the latter, the replacement of MAAs with flavonoids was an extraordinary molecular innovation, producing more than 9000 different flavonoid structures known to date. Modern land plants are capable of finely modulating flavonoid biosynthesis depending on the type and intensity of environmental stressors. In addition, the occurrence of flavonoids in different tissues and subcellular compartments well explains their ability to play multiple functions in response to environmental stimuli. In this review, we reasoned on the multiplicity of roles served by a nascent flavonoid metabolism during the “early steps” of plants in a land environment. Our reasoning stems from the notion that multi-functionality is a widespread property of most specialized metabolites, particularly flavonoids. Although these molecules undoubtedly allowed primitive plants to effectively deal with the most damaging solar wavelengths by acting as both UV filters and ROS scavengers, it is unlikely that these were their most prominent function in the process of terrestrialization. Indeed, we discussed that the benefits associated with the replacement of MAAs (excellent UV filters) with flavonoids mostly have involved the ability of these last to modulate phytohormone signaling; to assist the plant-fungus symbiosis, which is essential to plant development in the “new world” scarce of water and nutrients; besides offering effective defense against pest and pathogens.M. polymorpha and P. patens) have greatly advanced our knowledge about the molecular features of the evolution and the rise of specialized metabolic pathways that accompanied the plant terrestrialization. Despite the vast number of studies focused on angiosperms, very few have explored the effects of abiotic and biotic stressors on the biosynthesis of flavonoids in bryophytes, the earliest land plant group. Consequently, the functional roles of flavonoids in the interactions between early terrestrial plants and the environment often extrapolate from studies conducted on more complex plants. However, both the concentration in plant tissues and the inter- and intracellular location of flavonoids depends upon the evolution of the metabolism of these compounds and on the complexity of different land plants as well. This makes it difficult to assess the relative significance of the multifarious functions of flavonoids not only during the water-to-land transition but also in the success of plants in the conquest of strikingly different habitats. Therefore, this matter is critically important and merits further investigation.Recent studies on genome sequencing of both streptophyte algae and basal land plants (e.g.,


In an era of severe biodiversity loss, biological monitoring is becoming increasingly essential. The analysis of environmental DNA (eDNA) has emerged as a new approach that could revolutionize the biological monitoring of aquatic ecosystems. Over the past decade, macro-organismal eDNA analysis has undergone significant developments and is rapidly becoming established as the golden standard for non-destructive and non-invasive biological monitoring. In this review, I summarize the development of macro-organismal eDNA analysis to date and the techniques used in this field. I also discuss the future perspective of these analytical methods in combination with sophisticated analytical techniques for DNA research developed in the fields of molecular biology and molecular genetics, including genomics, epigenomics, and single-cell technologies. eDNA analysis, which to date has been used primarily for determining the distribution of organisms, is expected to develop into a tool for elucidating the physiological state and behaviour of organisms. The fusion of microbiology and macrobiology through an amalgamation of these technologies is anticipated to lead to the future development of an integrated biology. Populations are similarly undergoing marked declines, with losses in freshwater environments being considered the most serious.One of the most concerning global environmental issues confronting society today is the loss of biodiversity.Traditionally, the monitoring of organisms has involved the collection or observation of individuals and visual morphology-based identification. For the collection of aquatic species, methods such as fishing, electro-fishing, and the use of cast nets, gillnets, and trawls are routinely employed, whereas scuba diving and, more recently, underwater drones are used for visual observations. However, all these methods have the notable disadvantages of being labour-intensive and costly. Consequently, the monitoring of aquatic organisms to date has been far from comprehensive, whereas the loss of biodiversity continues at an unprecedented rate.sensu lato).In recent years, there has been a rapid development of environmental DNA (eDNA)-based analyses, in which information obtained by sampling DNA from environmental sources is used to identify the distribution and abundance of macro-organisms such as fish and amphibians. In this context, eDNA refers to all DNA in the environment, which, for example, includes both intra-organismal DNA of microorganisms and extra-organismal DNA derived from the faeces and mucus of macro-organisms in environmental media  naked DNA (dissolved DNA): DNA that has been released from cells and exists in a dissolved state in water; (ii) particle-bound DNA: DNA bound to particulate material, such as small mineral particles; (iii) intra-membrane DNA: DNA contained within cell organelles, intact cells, or parts of tissues. With respect to macro-organism-derived eDNA, particles trapped by filtration through filter paper with a mesh size of 0.45–1.5 μm are often used for analysis,Although analyses of macro-organism-derived eDNA typically target mitochondrial DNA for animals or chloroplast DNA for macroalgae and macro-plants, nuclear DNA is occasionally assessed.The origin of eDNA and the processes, by which eDNA released from organisms attains the aforementioned three states, are a particular focus of research; however, although a number of studies are ongoing, we currently lack a complete understanding. Taking fish as an example, the possible origins of eDNA include faeces, body surface mucus, sperm, eggs, and carcasses. However, there has been insufficient research to determine the proportional contributions of each of these sources to total eDNA.,Among the fundamental properties of eDNA, the most well studied is the process of degradation. In this regard, numerous studies have reported that eDNA undergoes exponential decay,,,The distribution of eDNA reflects the spatial distribution of its ‘owners’, and a number of studies have been conducted on the distribution range of extra-organismal DNA, albeit with contrasting findings. For example, studies on the downstream distribution of eDNA have tended to yield wide-ranging estimates from <100 mSince the publication in 2008 of the first paper describing estimates of organism distribution based on analyses of macro-organism-derived eDNA,Rana catesbeiana), an invasive species in Europe. Such methods are particularly useful for studying the distribution of rare or exotic species of interest, and have been used extensively, particularly in many of the early studies. Examples include the detection of endangered salmonids in US riversSpecies-specific eDNA analysis (eDNA barcoding) is an analytical method that focuses on a single species of interest. In the first documented example, Ficetola et al.,in silico confirmation and in vitro testing to ensure assay specificity. If, however, the database information proves insufficient, it is then necessary to determine the DNA sequences of the target species and its sympatric relative. Predictably, these processes are both time-consuming and costly, and when multiple species are targeted, these procedures need to be repeated for each target species. Having established these assays, however, the subsequent operations are relatively straightforward and inexpensive.The advantages of species-specific analysis are that it is simple, inexpensive, and relatively easy to prevent contamination. Most simply, the presence or absence of target species DNA can be determined by the presence or absence of an expected band using conventional polymerase chain reaction (PCR) and subsequent electrophoresis. In addition, as described below, this approach is compatible with quantitative methods, and it is relatively straightforward to determine the concentration of eDNA using real-time or digital PCR.,,,Trachurus japonicus) in the ocean. In this study, the numbers of Japanese jack mackerel in a bay estimated based on eDNA quantification and by echo sounder were almost identical, thereby demonstrating the feasibility of quantifying the absolute number of fish individuals using eDNA. However, the execution of this study required considerable effort, including modelling the flow field in the bay, using parameters of eDNA release and degradation rates obtained from tank experiments, as well as selecting a suitable season and an area with no appreciable variation in the size of the target fish species. This study accordingly highlights that whereas absolute abundance estimates based on eDNA quantification are achievable, we have yet to reach the stage at which we can readily determine the population density of a particular fish species by simply sampling water.As previously mentioned, it is possible to determine DNA concentrations for a given species in the environment using quantitative PCR methods. Positive correlations between eDNA concentrations in a particular water body and the abundance or biomass of organisms have been reported from the earliest days of macro-organism eDNA studies, not only in closed ecosystems such as lakes and ponds but also in open ecosystems such as lotic water bodies and oceans.,Exhaustive eDNA detection (eDNA metabarcoding) is a method that can be applied to comprehensively identify species in a certain ecosystem using universal primers to amplify the DNA of species belonging to a specific taxon, and then successively analysing the amplified sequences. This method, which at the earliest stages of development necessitated labour-intensive manual sequence reading,,The most advanced eDNA metabarcoding studies on macro-organisms conducted to date have been those performed for fish, for which a number of different gene regions have been used as markers for successful metabarcoding. Whereas early universal primers targeted CytB,Although apart from fish, comparatively few eDNA metabarcoding studies have been conducted for other taxa, in the case of amphibians, batra, a universal primer set for 12S rRNA, has been reported to significantly increase detection probability compared with sampling surveys,,,There have also been reports of the use of eDNA metabarcoding for the detection of terrestrial taxa,Although compared with quantitative PCR, metabarcoding is a less quantitative technique, efforts have been made to semi-quantify the analysis of eDNA in water. Ushio et al.,Cyprinus carpio) in Japan.Whereas the eDNA analysis described in the preceding section is primarily designed to achieve species-level resolution, if the resolution of analysis can be further enhanced, it would conceivably be possible to detect intraspecific polymorphisms, such as differences between local populations. In this regard, Uchii et al.Plecoglossus altivelis) from aquarium and natural river water.,A further example of higher resolution eDNA analysis is the detection of intraspecific polymorphisms using HTS targeting the rapidly evolving D-loop region of mitochondria, as exemplified by the successful detection of intraspecific polymorphisms in the ayu , the eDNA levels of which have been reported to increase in breeding habitats during the breeding season.,,,As externally fertilized animals, most fish species release large volumes of sperm into the environment during the breeding season, and given that sperm is among the sources of eDNA, it is predictable that the concentrations of such eDNA will increase during the breeding season. Examples include the hellbender  is believed to live for more than 100 yrs, a decline in the number of young individuals has been noted in recent years.In addition to assessments of organism status, methylation-related analyses may also enable age structure determination. In zebrafish, for example, it has been established that genome-wide methylation rates change with age,Earlier it was mentioned that the full length of the mitogenome is detectable in environmental water, and size fractionation revealed an abundance of eDNA larger than 10 μm in size.Recent technological advances have led to the rapid development of single-cell-based analysis methods, such as single-cell RNA-seq,In this review, I have touched upon the current status and future developments of eDNA analysis and related methods. eDNA-based monitoring has already become established as a ground-breaking novel technology, with significant implications for fields such as ecology. These developments have and continue to be made possible by techniques that have been developed in the fields of molecular biology and molecular genomics. It is anticipated that further advances will be founded on the integration of cutting-edge technologies that include more accurate long-read metabarcoding and environmental single-cell analysis. To date, microbiology and macrobiology have tended to develop independently with comparatively little cross-fertilization; however, technology-mediated fusion has the potential to bridge the gap between these discrete disciplines and thereby contribute to the development of an integrated biology.


The concept of cyberdelics emerged in the 1980s and 1990s as an umbrella term denoting the nexus connecting cybernetic  technologies and psychedelic (mind manifesting) drugs. Cyberdelic technologies, in particular the then newly emerging field of virtual reality, were touted by psychedelic cultural icons including Timothy Leary and Terence McKenna as auguring a new era of digital mind-expansion where psychedelic experiences will be recreated online inside virtual worlds. Cyberdelic culture waned in the 2000s. However, recent years have seen the return of the cyberdelic imaginary, following on the heels of a psychedelic resurgence and a renewed interest in virtual reality technologies and their use in therapy. Cyberdelic advocates speak of the necessity of creating transformative technologies that steer humanity away from mindless consumerism and distractedness, and towards expanded states of awe, presence, and transcendence. Nevertheless, much like psychedelics, cyberdelic technologies are seen as running against the grain of current sociocultural arrangements and economic models which threaten to quell their transformative potential. Research on psychedelics within the humanities over the past decade has emphasized the role of cultural set and setting: the significance of the cultural embeddedness of these psychoactive agents and the dependence of their effects on surrounding sociocultural conditions. Building on the notion of information technologies as mind-manifesting technologies, this paper sets out to consider what psychedelics can teach us about cyberdelics: how the principles of set and setting and current discussions within the psychedelic humanities can inform our understanding of the resurgence of interest in cyberdelic media, its prospects, and challenges. The theory that an alternative form of spiritual media can serve as an antidote to the maladies of alienated technological society and be key to solving global challenges may ring familiar to those acquainted with mid-twentieth-century psychedelic history. Back in the 1960s psychedelic advocates developed a vision of a society improved by the ingestion of psychoactives, which anthropologist Nicolas Langlitz terms a “political neurotheology” . PsychedAn updated version of the notion that technology may come to the rescue, enlighten and provide new horizons to a spiritually alienated humanity can be discerned in the concept of cyberdelics . CyberdeA distinct product of the West Coast and the ‘Californian Ideology’ that flaunted a mix of experiential libertarianism, entrepreneurialism, and techno-utopianism , the conThis perspective paper examines the cyberdelic idea: its cultural context, appeal, prospects, and challenges. By examining the story and ethos of cyberdelics as utopian, mind-manifesting technologies, it aims to illustrate the broader challenge of integrating mind-manifesting, psychedelic technologies within the context of contemporary high-octane, neo-liberal society. The endgame of this examination points to the inherent difficulty of modulating the nature of electronic (mind-manifesting) media within ingrained oppressive sociocultural conditions that favor dependent, compulsive patterns of media use. Within this context, cyberdelic media are in constant danger of being marginalized or repurposed to fit the values and norms of the surrounding society. The fate of these media therefore depends on their points of contact with broader sociocultural forces, including culture’s ability to allow for emergence of spaces of refuge from intense media intake, and its ability to imagine ways of being that are alternatives to the competitive, manipulation-oriented, consumerist matrix.The idea that digital technology may elevate human consciousness and assist humanity in realizing a loftier spiritual destiny may appear misplaced in the context of current discussions on the effects of media. The digital revolution, in particular its latest incarnations – the smartphone and social network revolutions – has been defamed in recent years. It is accused of wreaking havoc on human faculties of focus, concentration, empathy and connection, fomenting political polarization and unleashing an epidemic of mental illness . DigitalAnother dimension where digital technology’s effects are deemed to have been destructive is the domain of spirituality. Digital media are regularly described as robbing users of their sense of presence, bringing them into states of distraction, restlessness, fear of missing out (FOMO), anger, and envy that runs counter to spiritual ideals of presence, firmness and peace .Cyberdelic media present themselves as an alternative to the deleterious effects of contemporary digital media: media that cultivate harmony, awe, and presence rather than FOMO and dispersion, produce effects similar to those of the mystical-type experiences elicited by psychedelics, and are aligned with the aim of spiritual growth . The cybCyberdelic history has roots in late 1960s and early 1970s, at a time when psychedelics were having their cultural heyday, and the personal computer and internet revolutions were still nascent. At the time, the psychedelic movement cultivated a complex and often ambivalent relationship to technology. On the one hand, the movement was strongly aligned with the back-to-the-land commune movement, which cultivated naturalistic, frugal, low-tech, and even anti-tech ideals . SimultaIt was against this backdrop that computers began to be imagined as a form of mind-altering technology. In 1968 media savant Marshall McLuhan cryptically proclaimed that “the computer is the LSD of the business world, “hinting perhaps at the enhanced cognitive terrains opened up by computation , 83. In Over the next two decades, as digital technologies rose in prominence, psychedelics seemed to wane from the cultural horizon. Yet some digital visionaries held on to the hope that computers might hold the key to new mind-expanding breakthroughs. John Perry Barlow Grateful, a Grateful Dead lyricist turned digital ideologue, argued that virtual reality is the closest analogue to the psychedelic experience . VR pionTwentieth century cyberdelic culture peaked in the 1990s, alongside the internet revolution, and a small-scale psychedelic renaissance that emerged on the coasts of Goa and Ibiza, combining psychedelic compounds and electronic psytrance music designed to produce altered states of mind. However, the trend was short lived. While EDM and the internet revolution assumed a central cultural position, The VR winter of the 2000s, the fall of the Dot-com bubble, the corporatization of the web, the rise of digital surveillance, and other cultural factors such as the decline of the 1990s house/rave scene can all be surmised to have contributed to the waning of cyberdelic tide .Over the past two decades, a wave of techno-pessimism has all but completely reversed the socio-technical imaginaries around internet and information technologies. Digital technologies as such did not disappear from view, as can be recognized from the ascent of smartphones, social media and the cloud around this very time. Nevertheless, a radical shift has occurred in the ways such technologies are viewed and interpreted. Previously seen as technologies of individual empowerment aiding collective liberation, digital technologies came to be portrayed as inextricably enmeshed with the manufacture of addiction, corporate manipulation, political repression and a mental health epidemic .Rather than being seen as electronic psychedelics, digital technologies came to be viewed as analogous to narcotics and condemned as “electronic heroin”  and “digThen, in the mid-2010s something shifted again. Swept by the tides of a new psychedelic renaissance  and a viThe cyberdelic renaissance consists of a plethora of digital technologies purporting to support personal and spiritual growth. On one end of the spectrum, one finds relatively familiar, even prosaic, technologies such as smartphone apps that guide users through meditation and breathing exercises, or assist psychedelic users in the integration of psychedelic experiences . Other, The field’s growth has also been fueled by the rising interest in VR technologies and in VR therapy in particular. Recent years have seen growing interest in the use of VR in the preparation and integration phases of psychedelic therapy, and possibly during psychedelic sessions . The twoIsness purport to elicit empathy, ego dissolution and other Self-Transcendent Experiences (STEs) of the type induced by psychedelics by transporting users into a shared luminous virtual space where their beings are able to interact and even coalesce  makes evidently clear. However, the high-tech nature of the cyberdelic project is sometimes linked with a certain unease. Cyberdelic Society co-founder Carl Hayden Smith, for instance, proposes a human centered vision of hyper-humanism as an alternative to the technology-centered vision of transhumanism promulgated by techno-evangelists . Smith’sYet, beyond the question of supply chains, a more deeply felt suspicion of digital technology has been ingrained in contemporary society following revelations on the implication of such technologies in mass surveillance and election manipulation, and their disruptive economic, social and psychological effects . Recent Cyberdelic technologies offer hints that digital technology can be re-imagined. They provide evidence that computer technologies need not necessarily provoke FOMO, anxiety, and depression. Digital technologies can, under a different set of conditions, also produce experiences of awe, transcendence, empathy, and bliss. Still, the technological possibilities afforded by such media should not be conflated with social affordances, and herein lies the rub.Social Shaping of Technology (SST) theory argues that technological design and use are inextricably meshed with their surrounding society and culture. Technology theorists have incessantly pointed to the myriad ways in which technologies tend to assume the mores, values and beliefs of their time and place . ExaminiInterestingly, theories on the social shaping of technology resonate distinctly in one of the key concepts of psychedelic therapy and science: the idea of set and setting. Psychedelic compounds are regularly defined as mind-manifesting  non-specific, amplifying agents whose effects depend, first and foremostly, on their context of use . These apsychedelic; that is, they manifest the mind, “observes Erik Davis in his classic Techngnosis (Tying together SST theory and the idea of set and setting, it is worth noting that some authors have identified information technology as mind-manifesting technologies, namely exceptionally versatile and context-dependent. “Computers are literally hngnosis , 162. VRhngnosis , arguinghngnosis , 1, and hngnosis , 142. Cyhngnosis .Such proclamations regarding the context-dependency of digital technology provide valuable clues for the reasons digital technologies developed along narcotic, not cyberdelic, lines. As Yuval Noah Harrari opined regarding disruptive technologies of human-enhancement “If you have an arms race with these technologies then it’s very clear what kind of human abilities are going to optimized: it will be things like intelligence, it will be things like discipline … but other things like compassion, like artistic appreciation, like spirituality – these are things that are not very high on the list of armies and corporations. So the attempt to upgrade humans might actually result in downgraded humans.”.Digital media’s narcotic affordances of alienation and numbing should therefore be seen in the context in which they emerged. They fit the conditions of life in a competitive, hyper-individualist, neo-liberal culture characterized by boredom, a meaning crisis, and rising costs of living . Their rIs the quest for self-transforming and potentially society-transforming cyberdelic technologies inherently futile as long as society retains its current form? Folded in such discussions is one of the key questions in the theory of technology. Does technology shape society or does society shape technology?  While miThe challenges of cyberdelics therefore appear to mirror current conversations within the psychedelic humanities about the moral and political implications of the drugs . MedicalThe context of a cyberdelic resurgence is similarly crucial, though for different reasons. Products of deliberate engineered design, digital drugs are significantly more specific in their effects than the unruly psychedelics. The question here is whether a swing away from narcotic media and towards cyberdelic media is realistic given the socio-economic and cultural context of contemporary media use, and whether such media may earn their place in the competitive attention economy. While neo-liberal, free-market society continues to exert unabated pressures over a squeezed middle-class , the COVLike psychedelic neuro-theology, the cyberdelic project can be viewed as an attempt to invert the logic of media in order to foster alternative states of mind and shift the human condition characteristic of its era. For now, Cyberdelics remain an exotic form of alternative media which augurs exciting possibilities of undetermined prospects. Whether the project will ultimately come to be regarded as visionary or naïve will depend on the ways in which these mind-manifesting technologies come into contact with society’s cultural imagination and culture’s ability to imagine and support alternative ways of being outside the dopamine-addled consumerist matrix.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The author confirms being the sole contributor of this work and has approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Supercritical fluid extraction (SFE) has been recognized as the green and clean technique without any negative impact on the environment. Although this technique has shown high selectivity towards lipophilic bioactive compounds, very few case studies on the application of these extracts in final products and different food matrices were observed. Considering the recent developments in food science and the increasing application of supercritical extracts in meat products in the last decade (2012–2022), the aim of this manuscript was to provide a systematic review of the lipid extracts and bioactives successfully obtained by supercritical fluid extraction and their application in meat products as antioxidant and/or antimicrobial agents. Lipophilic bioactives from natural resources were explained in the first step, which was followed by the fundamentals of supercritical fluid extraction and application on recovery of these bioactives. Finally, the application of natural extracts and bioactives obtained by this technique as functional additives in meat and meat products were thoroughly discussed in order to review the state-of-the-art techniques and set the challenges for further studies. In 2020, modern meat market was USD 838 billion, and it is expected to further increase up to USD 1 trillion by 2025. During the same year, 328 million tons of meat were produced in the world, which is equal to 35 kg of meat/capita. Hence, meat market has large share in international food industry [Oxidative reactions (lipid and protein oxidation) and the growth and activity of microbial populations are the two main causes of deterioration in the quality of meat and meat products . Due to Depending on the initial hygiene and the preservation method used, the growth of microbial population (spoilage and pathogenic) can cause spoilage and accordingly various foodborne poisonings in meat and meat products . The groSalmonella spp., Escherichia coli O157:H7, Listeria monocytogenes) are primarily responsible for numerous foodborne diseases and food poisoning, which is why they are considered the most important limiting factor in the meat industry [The pathogenic bacteria  during processing, meat processors usually use numerous additives  . Howevern humans ,12. Thern humans . The natn humans ,8. The aMost research has focused on the use of medicinal and aromatic plants with proven antioxidant and antimicrobial potential ,15,16, bNutraceuticals and bioactive compounds can be isolated from natural resources by various extraction techniques before further use as natural additives in food matrices. Conventional extraction processes  can be effectively used for the recovery of lipophilic bioactive compounds. However, the use of these methods is decreasing, which is related to their limitations. The major problems with traditional extraction techniques are the use of expensive and toxic organic solvents , poor yield and quality of the final product, and huge consumption of time, energy and resources.The recent trend and challenges posed by green chemistry place high demands on chemical engineering technologies. The most important aspects have been defined by Chemat et al.  and are Salvia officinalis L.) herbal dust, where the SFE study started with the evaluation of the process kinetics [SFE has been particularly recognized as the green and clean technique with no negative impact on the environment. Moreover, this technique has shown high selectivity towards lipophilic bioactive compounds and the ability to obtain extracts with concentrated target compounds without traces of potentially harmful solvents. De Melo et al.  providedkinetics , determikinetics , which wkinetics , minced kinetics  and freskinetics .Considering the recent developments in food science and the increasing application of supercritical extracts, the aim of this manuscript was to provide a systematic review of the lipid extracts and bioactives successfully obtained by supercritical fluid extraction and their application in meat products as antioxidant and/or antimicrobial agents.Google.com, Bing.com, Yandex.com and Baidu.com. Time period was approximately set to 2012–2022 (accessed on 03.11.2021); however, more concern than time restraints was given to adequate coverage of the topics from the manuscript as types and origin of lyophilic bioactives, principles and parameters of SFE, and application of bioactives in meat products. This literature review focused on commonly used sources of scientific references as Web of Science, PubMed, Scopus, and ScienceDirect, together with usual internet search engines as Asteraceae, Lamiaceae, Lauraceae, Myrtaceae, Rutaceae and others  [The use of plant essential oils (EOs) has a long tradition in humankind, and they are usually obtained as secondary metabolites from various families of plants including itruses) ,28,29. Titruses) . ConductThe Eos and extracts are primarily composed of various groups of important compounds for meat industry due to their antimicrobial and antioxidative abilities, mainly referred to terpenoids , phenylpropanoids , carotenoids , tocochromanols (tocopherols and tocotrienols), and organosulfur compounds . Terpenoids are highly variable group of compounds also known as isoprenes, which are widely represented in plants while being formed from five carbon isoprene units. Polyphenols are secondary plant metabolites with minimum of one aromatic ring that has attached one (or more) hydroxyls groups . CarotenAs mentioned above, one of the main qualities for assessment of lipophilic bioactives for usage in meat industry pertains to their antimicrobial and antioxidant potentials . HoweverThymus serpyllum L.) applied to pork patties mainly contained terpenoids carvacrol (40%) and thymol (15–16%) with lesser quantities of α-terpineol, borneol and geraniol [Satureja montana L.) Eos that were used to extend the storage of fresh pork sausages mainly contained carvacrol (59%) with smaller amounts of thymol and polyphenolic 3,5-dimethylphenol. SFE extracts from the same plant also contained carvacrol (70.92% and 46.32%) where its content was strongly increased with higher pressure and temperature [Origanum vulgare) SFE extract contained tetrahydroxy-dimethoxyflavone derivative, rosmarinic acid, dihydroxy-trimethoxyflavone derivative, sakuranetin, velutin, luteolin-methyl ether, xanthomicrol, genkwanin, and diosmetin 7-O-rutinoside [Solanum betaceum Sendtn), due to its high content of polar polyphenols, is able to prevent lipid oxidation in cooked beef meat [Aside from terpenoids , polyphenolic compounds are another important group of major carriers of antioxidative activity in plant material. Main groups of polyphenols with desired qualities for meat industry include flavonoids, phenolic acids, stilbenes, coumarins, lignans, quinones, and curcuminoids . For insgeraniol . Furtherperature . Furthertinoside . It was eef meat . Rachis hypogea) and grapevines (Vitis vinifera), and Japanese knotweeds (Polygonum cuspidatum) is resveratrol. It is also present in red wine and mulberries (Morus). This chemical is also abundantly existent in grape pomace that is byproduct from winemaking industries , that possibly can increase meat quality  [Additional lipophilic polyphenol that originates from peanuts (el meat) . Similarel meat) .Bixa Orellana L.) [Solanum lycopersicum) or carrots (Daucus carota) and their byproducts. As expected, extracts with higher contents of carotenes will have lipophilic nature. With SFE of tomato byproduct it is possible to produce leoresin with high content of resin and essential oil that is rich in carotenoids and useful for the production of beef hamburgers [The next group of important lipophilic compounds originating from plant material are carotenoids (largely used as colourants and antioxidants) ,43. One lana L.) . Bixin alana L.) . It was Tocochromanols are important antioxidants and vitamins that are almost equally interesting for human health and food production. Vitamin E is one of the lipophilic tocos that is a fat-soluble molecule with eight different structures, where four of them are tocopherols and another four are tocotrienols . Raw matAllium sativum) and other allium members are rich with lipophilic organosulphur compounds with antioxidant and antimicrobial qualities [The Eos made from garlic  using organic solvents such as toluene, hexane, heptane, petroleum ether, dichloromethane, chloroform, ethanol, methanol, etc., is a traditionally accepted procedure for the recovery of various classes of lipophilic compounds. This process is usually carried out at lower temperatures than conventional hydrodistillation (HD), thus reducing the risk of possible chemical changes and/or degradation of target compounds due to elevated temperatures. Although non-polar organic solvents have good affinity and selectivity for lipophilic bioactives , concomitant lipophilic compounds such as waxes may be co-extracted, reducing the content of target compounds in the extracts. The use of organic solvents in SLE is associated with other major problems such as particularly high costs due to the consumption of expensive resources, poor environmental impact due to the discharge of the chemicals used, and the major risk of human toxicity that may occur during production or due to the traces of the solvents in extracts and final products.On the other hand, HD is the most commonly used method for isolating non-water-soluble compounds with high-boiling points such as Eos, which consists only of volatile compounds. This technique is the most widely used in the world due to its simplicity, it is considered the most cost-effective technique for the production of most Eos at a reasonable price in less developed countries . AlthougSFE has been developed as an environmentally friendly and clean technique that can overcome the above challenges associated with SLE and is an excellent alternative and a current method of choice for the isolation of lipophilic compounds. Over the last 40 years, this technique has become increasingly important in the food, pharmaceutical and cosmetics industries. The most important aspects of these improvements over SLE and HD are reduced solvent consumption, preservation and reuse of energy, reduced time, improved selectivity and total yield, and avoidance of degradation of bioactive compounds by high temperatures.2 has other desirable properties as it is non-flammable, non-explosive, available in high purity, relatively cheap, non-toxic  and inert [The supercritical state of matter is associated with a tremendous effect on its physico-chemical properties, which is the main principle of SFE. For example, the increase in pressure leads to an increase in the density of supercritical fluid, whereas some of its transport properties  are more similar to those gasses . The lownd inert , which mExtraction and separation are the two main steps of the SFE process. The solvent is first compressed to the working pressure and temperature, which allows its diffusion into the plant matrix and mass transfer from the solid to the liquid phase . The sepEfficiency, yield and selectivity of the SFE process depend largely on the choice of the main process parameters . Therefore, the application of SFE requires the use of a very specific high pressure process equipment with integrated control of the factors affecting the process. SFE parameters must be selected according to the physicochemical properties of the target compounds and the process should be frequently optimized for each case study to achieve maximum yield of the target compound(s). Solvent solubility is directly affected by pressure and temperature; therefore, they are often considered the most important SFE parameters. Increasing the solvent density and thus improving the solubility is associated with increasing the pressure. This leads directly to a higher total extraction yield, which in certain cases can represent a significant improvement in process efficiency. However, the increased solubility leads to the co-extraction of other lipophilic compounds. For example, an SFE extract of coriander obtained at 100 bar and 40°C would be rich in volatile terpenoids , whereas SFE at 300 bar and 40°C would tremendously improve the total yield due to co-extraction of fatty oil, resulting in low selectivity towards terpenoids . This le−3 of supercritical carbon dioxide, making it a suitable solvent for the isolation of lipid compounds. Understanding the thermodynamic and kinetic nature of the SFE process is critical to ensure high selectivity of extraction and reduce the possibility of co-extraction of non-target compounds [Contrarily to pressure, an increase in temperature has a negatively effect on solvent density, often resulting in poor extraction yields. However, temperature affects both solvent properties and vapor pressure of solutes, which could improve their solubility and lead to higher yield . Since tompounds . 2 with the addition of ethanol as a co-solvent for the preparation of anthocyanin-rich extracts, which have been discussed elsewhere [For example, EO terpenoid-rich extracts may often be contaminated with other non-volatile lipids that could affect the bioactivity of the extract and degrade its quality. It should be emphasized that the selectivity of supercritical carbon dioxide is limited to different classes of lipid compounds and is very limited for the recovery of polar and moderately polar bioactives. The selectivity and solubility of supercritical carbon dioxide can be modified by adding various co-solvents, which can be either polar (ethanol and methanol) or non-polar (hexane and methylene chloride). Since non-polar organic solvents have low selectivity, their use as co-solvents could have a positive effect on the total extraction yield. However, their use is not often recommended because it affects the green character of the SFE. On the other hand, ethanol is most commonly used as a co-solvent for SFE due to its low miscibility with carbon dioxide, lower toxicity and easy removal from the extract . This aplsewhere .In addition to pressure and temperature, solvent flow rate and extraction time are also studied as important parameters of SFE. Mass transfer parameters such as axial dispersion, convective mass transfer coefficient and accumulation of extracted compounds in the supercritical phase are highly influenced by the flow rate of solvent . IncreasTime is also an essential attribute of any technological process. Increasing the extraction time would usually lead to an asymptotic increase in the total extraction yield, and time is closely related to solvent consumption. Therefore, optimization of SFE can be performed by selecting the total extraction yield or the yield of target compounds as the responses . Most stThe application of SFE for the recovery of lipophilic bioactives is associated with severe drawbacks, as the processes must be performed at high pressure to maintain the solvent in a supercritical state. Compared with conventional SLE methods, this directly requires higher operating costs . SFE reqMentha piperita L. (mentha), Rosmarinus officinalis L. (rosemary), Lippia thymoides L. (lippia), and Origanum vulgare L. (oregano), leaves and flowering tops from Ocimum basilicum L. (basil), aerial parts of Satureja montana L. (winter savory), seeds of Coriandrum sativum L. (coriander) and Piper nigrum L. (pepper), rhizomes from Zingiber officinale L. (ginger), flowers of Lavandula angustifolia L. (English lavender), peels of Citrus sinensis L. (orange) and Citrus tangerine L. (tangerine) or fruits of Carum copticum L. (ajwain) and Malpighia Punicifolia L. (acerola)  .Salvia officinalis L. (sage) and Thymus serpyllum L. (wild thyme), which is not suitable for the production of filter tea bags due to the low-particle size of these fractions, is discarded as a by-product [The raw material for the extraction of lipophilic compounds may be plants grown specifically for essential oil production, or by-products of the tea or fruit processing industries. For example, orange and tangerine peels left over after juice processing could be used to produce extracts enriched in terpenoids and fatty acids ,77. Addi-product ,36. Howe2 pressure, which is one of the most important SFE parameters, was set at 71 bar to recover 2-furancarboxaldehyde,5-(hydroxymethyl) and other compounds from acerola [The applied SFE parameters for the recovery of lipophilic compounds can vary to a great extent. For example, CO acerola , whereas acerola . Tempera acerola . The hig acerola ,66. The time taken to extract the plant material may also vary. A total of 40 min (dynamic stage) was required to isolate an extract containing 25.38% β-caryophyllene, 15.64% limonene and 13.63% sabinene in volatile phase from pepper . On the Similar to the total extraction yield, the chemical composition of the lipophilic extracts correlates strongly with the plant material, but also with the SFE parameters. Carvacrol and/or thymol, which are cyclic monoterpenoids and derivatives of p-cymene, can be found in significant amounts in oregano leaves , ajwain Mattje et al.  reportedThe main studies conducted on the application of natural extracts obtained by SFE in meat and meat products are summarized in Echinacea spp. is a well-known medicinal plant from North America with strong antioxidant potential [Echinacea extracts were acceptable to consumers. otential . Gallo eotential  investigOriganum vulgare L.) is used in food industry as a flavouring agent and natural additive because of its flavouring properties, which are due to its essential oil. Oregano essential oil is mainly rich in terpenoid compounds , which have obvious preservative potential in various foods [Oregano  is known as a medicinal and aromatic plant with high content of bioactive compounds such as rosmarinic acid, carnosic acid and carnosol, which have significant antioxidant, antiviral, antibacterial and anti-inflammatory potential [Rosemary  to reduce cholesterol oxidation. The results of this study indicate that the addition of oregano and rosemary extracts (1 and 3 g/kg) effectively reduced cholesterol oxidation products  in fish patties during 14 days of cold storage . Salvia officinalis L.) has long been used as a medicinal and aromatic plant, for its specific flavour and its strong antioxidant and antimicrobial effects. The predominant compounds in sage essential oil are monoterpene ketones, although its strong antioxidant potential is mainly associated with the presence of diterpene polyphenols [Listeria monocytogenes and Escherichia coli) in ground pork [E. coli in ground meat during 8 days of cold storage for. In another study, Danilović et al. [L. monocytogenes and, consequently extend the shelf-life of ground meat by up to 6 days.Sage  has a high content of bioactive compounds, including gingerol and its derivatives, with a strong antioxidant potential [Ginger  is known as a medicinal plant with diverse pharmacological activities due to its chemical form. The most abundant compounds in the essential oil of winter savory are phenolic terpenoids, thymol, and carvacrol, which have potent antioxidant and antimicrobial potential [Winter savory  are characterized by the presence of various types of bioactive compounds  that have significant antioxidant, antitumor, antimicrobial, and hypoglycemic potential [Enterobacteriaceae). The strong preservative effect of these extracts could be related to their chemical form: high content of carvacrol and thymol.Extracts of wild thyme  is an indigenous fruit from South America with significant content of bioactive compounds, including carotenoids, anthocyanins and other phenolic compounds with important preventive, biological and therapeutic properties [2 and the addition of EtOH as a co-solvent. The use of EtOH as a co-solvent resulted in an increase in the content of polar phenolic compounds with strong antioxidant potential. Berries are predominantly rich in phenolic compounds with strong antioxidant potential, and therefore the use of various fruit extracts in meat products suggests good antimicrobial potential . Tamariloperties . Castro-operties  studied operties  found thMalpighia punicifolia), long used in folk medicine, has been studied in vitro for its antibacterial activity. Tremonte et al. [Brochothrix thermosphacta and Pseudomonas spp. in water buffalo steaks at concentrations of 0.0063 and 0.0125% and significantly reduced the initial cell count at the concentrations of 0.025 and 0.05% [The extract of acerola  peel is a major by-product of fruit processing with numerous health benefits: antioxidant, anticarcinogenic, antibacterial, anti-inflammatory, and antiatherosclerotic activities. The bioactive potential of pomegranate peels might be related to the high content of polyphenols, especially ellagitannins [2 was performed. Two fractions of pomegranate peels were prepared: a fraction with high phenolic content and a lipid fraction. The results of this study showed that the phenol and lipid rich fractions from pomegranate peels effectively reduced lipid and protein oxidation without negatively affecting the colour changes of the fish pies. It is also worth highlighting that both fractions have higher antioxidant potential than the synthetic antioxidant butylated hydroxytoluene. The obtained results suggest that pomegranate peel fractions could be used as an effective substitute for synthetic antioxidants in fish meat. Pomegranate  and from berry fruits  obtained by SFE in meat processing. The results presented in this review indicate that the extracts obtained by SFE can be successfully used as novel antioxidants and antimicrobial agents in various types of meat and meat products. In some studies, it was found that the use of SFE offered higher efficiency and better selectivity, and thus better antioxidant and antimicrobial potential of the newly formed extracts than conventional methods, including hydrodistillation and Soxhlet extraction. Further studies should evaluate the ability of the natural additives obtained by SFE as potential substitutes for sodium nitrite in cured meat products.Although the interest of the scientific community in the study of natural extracts obtained by SFE is constantly growing, there is still room for further improvement to expand the applications in the industrial sector. Therefore, further investigations and challenges can be divided into several directions. First, research should focus on the production of high-quality extracts rich in bioactive compounds  by SFE. In addition to innovating the SFE process and optimizing the extraction procedure, alternative plant resources  should be used as raw materials for the isolation of bioactives. Another important aspect in the production of these extracts is the encapsulation process to find a suitable way to protect the bioactive compounds from possible degradation and to ensure high quality of the final product in terms of odor, solubility of bioactive compounds and improved efficacy (better bioavailability). Finally, it is expected that the number of studies dealing with the application of extracts obtained by SFE as natural additives in meat products will increase and will be studied in different meat products, as well as in different food matrices.