Oral health is an underestimated factor affecting overall human health and quality of life. Long-term enteral or parenteral nutritional treatment requires not only regular assessment of access routes, the patient’s nutritional status, and tolerance to the selected method of nutrition but also of oral health. This article discusses the connections between the influence of chewing function, salivation, and xerostomia on the health of the oral cavity of patients on long-term enteral and parenteral nutrition. In addition, the role of nurses in assessing oral health is presented as well as crucial elements of a comprehensive oral assessment in a nursing care plan. Patients receiving long-term enteral and parenteral nutrition have an increased risk of developing oral diseases. Increasing knowledge about the factors affecting oral health among nurses is crucial to provide appropriate care for patients requiring long-term nutritional treatment with omission of the natural route of food intake. Regular assessment of oral health by nurses should be an important aspect in long-term nutritional treatment recommendations. The assessment and care of the oral cavity has been an important element in the care of patients in intensive care units. Its effectiveness in preventing ventilation-associated pneumonia (VAP), aspiration pneumonia, or hospital-acquired pneumonia (HAP) has been confirmed by many clinical studies . A recenConcern for oral health is also very important in the care of patients receiving long-term enteral and parenteral nutrition. Enteral nutrition (EN) therapy has long been recognized as the standard in nutrition treatment and is an invaluable alternative in the case of significant difficulties in naturally consuming food through the mouth. The supply of nutrients to the gastrointestinal tract requires the placement of a feeding tube or catheter or creating a stoma. The greatest benefit of enteral feeding is the maintenance of gut stimulation and integrity . ParenteEnteral or parenteral nutritional treatment can be provided both in the short and long terms. The administration of enteral or parenteral nutrition for more than 6 months is defined as long-term medical nutritional therapy .The omission of the natural route of food intake not only deprives the patient of the pleasure of eating but is also associated with the risk of complications and accompanying unpleasant sensations. Impaired masticatory function, regardless of the cause, is an important factor causing oral health problems, and patients, as well as their caregivers, are often unaware of the importance of oral health as an important component of overall health . Lack ofExisting recommendations for long-term enteral and parenteral nutrition address many aspects of nutritional therapy, but none of the documents address oral hygiene or the systematic assessment of oral health ,10,11. SThe aim of this study was to raise awareness of the importance of assessing the oral health status of patients on long-term enteral or parenteral nutrition, to highlight the role of the nurse in this regard, and to discuss selected factors such as chewing stimulation, salivary secretion, and xerostomia, which can have a significant impact on oral health in this patient group.Oral health is an important part of a person’s physical and mental health. It has been shown to have a significant relationship with many chronic diseases and can affect dietary choices, ability to communicate, and functioning in society . Oral heIn the long-term parenteral nutrition patient population, oral health status is of significant importance and can affect overall outcomes. A prospective study with 52 home parenteral nutrition patients revealed that the accumulation of multiple risk factors, such as chronic malabsorption, dehydration, multidrug therapy, and symptoms of xerostomia, combined with nutritional therapy, increased the incidence of dental disease and oral infections. A total of 17 (33%) of the patients experienced a change in their previous pattern of dental attendance, with 14 (27%) attending less frequently and 3 (6%) attending more often . FurtherThe profile of patients requiring enteral and parenteral nutrition has changed over the years. They tend to be older patients, over 65 years of age, and have serious comorbidities . Aging aLactobacillus spp., Staphylococcus spp., Streptococcus spp., Bifidobacterium spp., or Gram-negative rod-shaped bacteria of the Enterobacterales. With time, the human bacterial flora begins to stabilize, and its composition depends on exogenous factors, such as living conditions and the type of food and endogenous factors, such as health or genetic conditions [The human oral cavity is not a homogeneous ecosystem. The oral microflora begins to form during early childhood and changes throughout life. In the early neonatal period, a large part of the oral microflora comes from the mother, e.g., nditions . BacteriThe flora that colonize the oral cavity include both commensal and pathogenic bacteria. Adequate oral hygiene and a balanced diet help to maintain the balance between microorganisms. Changing any factor may create an environment in which the balance of the number of microorganisms is disturbed, which results in the multiplication of pathogenic bacteria. Elderly patients with limited chewing and swallowing ability, ill-fitting dentures, or poor oral hygiene are particularly exposed to oral dysbiosis processes .Lactobacillus salivarius, which can inhibit the colonization of Helicobacter pylorii in the stomach. They suggested that administering L. salivarius can prevent dental caries and periodontitis [P. gingivalis can lead to the destruction of the intestinal barrier and trigger a systemic inflammatory response and aggravate other systemic diseases [Oral food intake plays an important role not only in nutrition but also in maintaining the microbiological balance in the complex oral and gut ecosystem. Katagiri et al. observed that the reinitiation of oral nutrition for poststroke patients significantly changed the composition of the oral microbiota, but not that of the gut microbiota. They found that there was a change in the interaction between microbiome species and an increase in the activity of dontitis . Parentedontitis . Microbidiseases .Chewing is the first and crucial step in digestion and requires the coordination of the tongue, facial muscles, jaw, and teeth. It is an important process not only because of food intake: the masticatory apparatus is significantly related to the process of speech production and has an impact on general health. Chewing stimulates the production of saliva, which indirectly influences digestive functioning . In the case of enterally and parenterally fed patients, maintaining the masticatory apparatus in good condition is of great importance. The inability to orally take food or insufficient food intake may result in higher rates of caries, decreased antimicrobial activity of the saliva, drying out of the oral mucosa, adherence of thick sputum to the palate, and neglecting oral hygiene during periods of underlying disease activity .Masticatory performance problems have been associated with loss of teeth, wearing removable dentures, reduced salivation, tongue disease, loss of muscle mass, and increased nutritional deficiencies . TherefoSaliva is a dilute aqueous solution that consists of organic and inorganic components and plays an important role in maintaining the microbiological balance in the mouth. The high water content of saliva helps to moisturize the food and makes it easier to chew. The secretion of saliva is controlled by the autonomic nervous system, and the amount of saliva produced depends on the type and intensity of stimulation. Under physiological conditions, an adult produces 0.5 to 1 L of saliva per day. The main stimulator of salivation is food, and much larger amounts are produced before, during, and after meals. During sleep, physiological saliva secretion is reduced . Based on the physiological characteristics of the functioning of the salivary glands, saliva has many important functions, such as protecting the teeth and soft tissues against mechanical, biological, and chemical factors. Continuous moistening of the oral cavity is of key importance for speech and taste and supports the processes of chewing and swallowing .Xerostomia is associated with reduced saliva secretion and a change in its chemical composition, resulting in a dry mouth. The classification includes xerostomia vera (primaria), which results from disturbances in the functioning of the salivary glands and manifests itself as a group of related symptoms; and xerostomia spuria (symptomatica), which is characterized by a subjective perception of dry mouth, without any objective signs of dysfunction of the salivary glands . Most people have experienced or experience this feeling every now and then, especially in stressful situations; however, this is usually a short-term discomfort. Chronic xerostomia is a much more serious problem, as it may affect speech, cause swallowing difficulties, taste disturbances, mechanical damage, or chronic oral infections, and result in the need for dentures, affecting the overall quality of life . Xerostomia is more commonly reported in people over 65 years of age. The occurrence of xerostomia is strongly associated with comorbidities, polypharmacy and disease treatment, smoking, or inflammation of the salivary glands. Persistent dry mouth may accompany diseases such as type I diabetes, hyperthyroidism, neurosis, schizophrenia, depression, Parkinson’s disease, Alzheimer’s disease, Addison–Biermer anemia, Sjögren’s syndrome, as well as hypercalcemia or avitaminosis . Chronic anxiety and stress also impair the functioning of the salivary glands . The long-term use of EN and PN indirectly influences the environment and condition of the oral cavity. Malnutrition and dehydration, common among patients with inflammatory bowel disease or bowel failure after extensive resection of the small intestine, also reduce salivation. Lee et al.  found thChronic dry mouth has an important psychological aspect, as it significantly affects the quality of life of patients. Therefore, the diagnosis of dry mouth requires a detailed clinical anamnesis, including comorbidities, medications taken, the course of therapies  of the underlying disease, and the assessment of objective symptoms reported by the patient . A summaMore than 100 questionnaires, available in the literature, have been developed for assessing the severity of dry mouth and salivary gland functioning. The most common questionnaires used for the subjective diagnosis of xerostomia are: Xerostomia Inventory (XI), Fox’s Questionnaire, Shortened Xerostomia Inventory (SXI), and Visual Analogue Scale (VAS) . AnotherIn the recent Global Oral Health Status Report, the WHO highlighted that close to 3.5 billion people worldwide suffer from oral diseases . AccordiUpdating interprofessional knowledge in the field of oral hygiene is an indispensable element of providing comprehensive nursing care for the patient, as is cooperation with the family in effective prophylaxis.The professional preparation of the patient and their caregivers to take action to promote oral health must be based on a comprehensive assessment of the patient’s situation and individual needs. The patient’s oral health assessment steps are shown in Therefore, the role of specialists other than dentists, such as nurses, is growing in the field of screening and early detection of oral health problems. Kohli et al.  reportedThe nursing assessment of oral health should be individualized. This is a very important approach in health care, where patient satisfaction determines the quality of care. Specifically, the nursing care plan for long-term enteral and parenteral nutrition patients should include an oral health assessment, which consists of the following areas and abilities: the patient’s health situation, oral health status, self-care ability, cognitive ability, patient/caregiver knowledge of factors that positively and negatively affect oral health, and preparation of an individualized oral care plan. The assessment categories and objectives are described in The oral health of patients on long-term enteral and parenteral nutrition requires proper attention. Patients’ ability to maintain their oral health may be limited. The lack of stimulation of the masticatory organ, dry mouth, and inadequate care can cause significant changes in the oral microflora and increase the risk of developing serious diseases in this area. Providing quality oral health care is a major challenge for the healthcare system. A better understanding of the factors affecting the oral health of patients requiring specialized nutritional treatment is elementary in nursing practice. Nurses, in providing care to different groups of patients, play a significant role in educating and promoting oral health. They can perform oral health assessments, train and manage the activities of care assistants, and educate patients and their families. The role of nurses in oral assessment is growing, but is still insufficient. In addition, caring about oral health is still a low priority in society. Nurse education in this area should be promoted, and the priority of care for maintaining oral health in patients with enteral and parenteral nutrition should be raised. Comprehensive care for enterally and parenterally fed patients must take into account oral care, especially when disease is active.Patients in the hospital and in primary care should be supported by nursing and caring staff trained to accurately recognize symptoms of poor oral health and implement appropriate measures.An oral health care assessment should be a standard examination in long-term nutritional therapy.The implementation of professional training for nurses in the field of oral cavity assessment and care would improve the activities promoting oral health among adult patients treated with enteral and parenteral nutrition and other groups of chronically ill patients.


Chronic wounds are becoming an increasingly common clinical problem due to an aging population and an increased incidence of diabetes, atherosclerosis, and venous insufficiency, which are the conditions that impair and delay the healing process. Patients with diabetes constitute a group of subjects in whom the healing process is particularly prolonged regardless of its initial etiology. Circulatory dysfunction, both at the microvascular and macrovascular levels, is a leading factor in delaying or precluding wound healing in diabetes. The prolonged period of wound healing increases the risk of complications such as the development of infection, including sepsis and even amputation. Currently, many substances applied topically or systemically are supposed to accelerate the process of wound regeneration and finally wound closure. The role of clinical trials and preclinical studies, including research based on animal models, is to create safe medicinal products and ensure the fastest possible healing. To achieve this goal and minimize the wide-ranging burdens associated with conducting clinical trials, a correct animal model is needed to replicate the wound conditions in patients with diabetes as closely as possible. The aim of the paper is to summarize the most important molecular pathways which are impaired in the hyperglycemic state in the context of designing an animal model of diabetic chronic wounds. The authors focus on research optimization, including economic aspects and model reproducibility, as well as the ethical dimension of minimizing the suffering of research subjects according to the 3 Rs principle . Epidemiological data clearly show that diabetes mellitus (DM) has long been a challenge for the representatives of the medical world and healthcare-financing entities. Estimates show that the number of DM cases and diabetic complications worldwide is increasing, including difficult-to-heal wounds, with the most representative example being diabetic foot syndrome (DFS). It is assessed that during the lifetime of a patient with DM, the probability of DFS occurrence ranges from 5 to 25%, according to various authors. DFS is found in 10 to 20% of patients with diabetes, and detailed data from the USA report even 25% . ConsideThe source of impaired wound healing in DM is related to several overlapping mechanisms, presented in Endotheliopathy plays a leading role due to impaired tissue metabolism in DM . It is r− and O2−. The formation of ROS products exacerbates endothelial cell dysfunction, thus mediating DNA damage [The polyol pathway leads to endotheliopathy through the decreased supply of NO in dysfunctional vessels by reducing NADPH in the cytosol. NADPH is necessary for the regeneration of glutathione, which has antioxidant properties, and also for the production of nitric oxide synthase (NOS) . In the A damage . IncreasA damage . Not onlA damage . These pA damage . A studyA damage . Animal Most of the processes responsible for the development of endotheliopathy, including increased polyol pathway activity, AGE formation, and the PKC pathway, simultaneously lead to the development of neuropathy . In addiBesides autonomic responses, unmyelinated C-type fibers are responsible for pain perception. Up to 70% of multifunctional C-fibers are present in the skin ,33. TheiImpaired wound healing in the course of ulceration results most often from an ongoing infectious process. A typical feature of chronic wounds is an increased concentration of metalloproteinases, which causes damage to the formation of granulation tissue . A chronThe nature of atherosclerotic lesions in DM results not only from their much faster development compared to the general population but also from a subsequent predilection for lesions to be located in the intima and the media . The dif2− when treated with high concentrations of glucose [An impaired activation of many overlapping processes is the molecular basis of such a phenotype of changes as the ones observed in DM. Diabetic endotheliopathy is the initial process that facilitates the formation of vascular stenosis that initiates atherosclerotic plaque formation. Endothelial dysfunction is initiated not only by hyperglycemia but also by other factors such as increased arterial pressure, causing turbulent flow in vessels, high concentration of oxidized LDL, high concentrations of free oxygen radicals as well as homocysteine, genetic factors, inflammatory changes resulting from viral or bacterial infections, and the response to proinflammatory cytokines . One of  glucose . This ph glucose . The rol glucose . The adhesion of platelets to damaged endothelial cells is one of the factors enabling the formation of atherosclerotic plaque and embolism. In DM, this process is enhanced by increased stimulation of growth factors, fibrin, thromboxane A2, integrins, and P-selectin. Activated platelets secrete numerous molecules which maintain local inflammation by stimulating endothelial cells. This leads to a constant mutual reaction between the endothelial cells and the atherosclerotic plaque, including the transformation of monocytes into macrophages by enhancing diapedesis . Moreover, according to the current views, this subclinical inflammation is responsible for initiating and maintaining the atherosclerotic process. Indeed, there is a relationship between the activity of the innate and adaptive immune responses and the propagation of atherosclerotic lesions, which is particularly seen in DM. The innate immune response is activated by the subendothelial accumulation of cholesterol deposits and foam cells (fat-laden cells with an M2 macrophage-like phenotype), oxidation, and glycation ,54,55. IIn DM, chronic inflammation is responsible for the long-lasting wound healing process, which is another factor stimulating the development of atherosclerotic lesions. The activation of Toll-like receptors (TLRs) in the damaged tissue that are part of the innate immune system is the phenomenon linking these two processes . During Pentraxins  constitute another group of factors that link the atherosclerotic process and the wound healing process. PTX3 is synthesized during neutrophil differentiation, it is accumulated in their mature follicles, and is ready for secretion under hypoxia in response to increased concentrations of proinflammatory cytokines such as TNF alpha, IL-1 beta, or the presence of lipopolysaccharides ,70,71. TAdditional factors linking the pathogenesis of atherosclerotic lesions and the healing process are cathepsins and matrix metalloproteinases (MMPs), whose dominant role is the degradation of extracellular matrix components. Under physiological conditions, they are responsible not only for the reconstruction of vascular wall structures as well as formation and destabilization of the atherosclerotic plaque, but also for the regeneration of superficial injuries of the epidermis or mucosa ,76,77. KFurthermore, cathepsins play an essential role in wound healing. Depending on their site of action, they affect not only cardiac remodeling (cathepsins B and S) but also bone (K) and adipose tissue (S) metabolism ,90. MoreThe formation of atherosclerotic lesions would not be possible without the influence of procoagulant factors. Activation of the plasmin system appears to be particularly important in DM ,96. PlasMast cells play a special role among inflammatory cells involved in the development of atherosclerosis. They mature in the vessel wall under the influence of stem cell factor (SCF). They produce and secrete proteolytic substances which play a significant role in vessel wall remodeling. These proteolytic enzymes modify the composition and function of LDL and HDL lipoproteins . In rodeMast cells, which secrete paracrine and autocrine proinflammatory cytokines such as tumor necrosis factor (TNF), IL-33, and IL-6, participate in the mechanism of insulin resistance by activating kinases . IL-37  and class B scavenger receptors (CD36). The role of these receptor types is to engulf modified lipids in the vessel wall for clearance. CD36 appears to be more important in activating monocytes and macrophages in the adipose tissue, liver, and arteries, leading to increased levels of soluble sCD36 in plasma and consequently maintaining the atherosclerotic process ,108. IncMoreover, expression of the transmembrane lectin-like oxLDL receptor 1 (LOX-1), which is responsible for trapping oxidized LDL, has been found in endothelial cells, vessel wall smooth muscle cells, platelets, and macrophages . The conWound progression and healing in DM in humans is a complicated process, as has previously been described in detail. As a result, the design of appropriate and potentially successful therapies requires optimal preclinical models of these processes. Several animal models have already been created, such as mouse, rat, hamster, and pig models (widely used models are given in In an established pig model (streptozotocin-injected Yorkshire pigs) of diabetic wound, the wounds healed within 18 days, which makes this model incoherent with the analogous process in humans . StreptoA study conducted by the authors of this review showed that the duration of hyperglycemia in rats might reflect the long-term uncompensated course of DM in humans . The resOf note, the chronic wound model described in the vast majority of studies is in fact an acute wound model. One example is the rabbit model, in which two of the three arteries supplying the area with blood were ligated in the rabbit’s ear . Next, aAnother concept for model selection is the anatomical recreation of the human skin model of a chronic wound in pigs . As in tMost current animal models are based on the formation of acute wounds and then subjecting the wounds to factors responsible for the formation of chronic wounds, i.e., ischemia  or DM. In the above study, a model imitating chronic wound conditions in patients with DM was presented . The preAnother modification in out abovementioned study was the use of bacterial lipopolysaccharide placed directly on the wound as an additional factor for improving the model to recreate the conditions in the human model. The presence of DM in humans is a widely recognized factor for an increased risk of wound infection through hyperglycemia and also macro- and microangiopathy, causing worse tissue perfusion, as well as autonomic neuropathy, which is the cause of excessive drying of the skin and, as a consequence, its poorer regeneration. In their meta-analysis, Biancari and Giordano put forward a thesis about the existence of a positive correlation between the worse metabolic control of DM manifested in objective laboratory tests as a higher percentage of glycated hemoglobin and the frequency of postoperative complications in the form of wound infection after cardiac surgery . For thiIn the above paper, special modifications were applied to the model after the pilot study . This inAnother concept for choosing a chronic model is the wound model presented by Reiss et al., who used a special Xanthan gum carrier applied to the wound. Recombinant collagenase  was suspended in Xanthan gum to a concentration parallel to that found in chronic wounds . In the As mentioned before, the species of animals present different advantages and disadvantages in an animal model of diabetic wounds. The selection of the appropriate strain is also important. For instance, Zucker Diabetic Fatty (ZDF) rats or Zucker-Diabetic-Sprague Dawley rats are more specific for investigation of type 2 DM than Wistar-Kyoto (WKY) rats (used in studies on hypertension) ,138,139.The risk of a wound in a diabetic patient lies not only in the fight against potential infection but above all in its long-term persistence, which, with a decreased immune barrier, exposes the patient to life-threatening complications such as sepsis. Patients with DM are also at greater risk of wound tissue resection due to severe infection or irreversible ischemia of the wound area. Since consequences such as high amputation significantly shorten the life of patients with DM, intensive treatment is needed as early as possible in the course of the disease to prevent its chronicity . Hypergl


GNAS mutations are the most prevalent cause of somatotroph tumors, the cause of half of all pathogenesis occurrences remains unclarified. However, recent findings including the pangenomic analysis, such as genome, transcriptome, and methylome approaches, and histological characteristics of pituitary tumors, the involvement of AIP and GPR101, the mechanisms of genomic instability, and possible involvement of miRNAs have gradually unveiled the whole landscape of underlying mechanisms of somatotroph tumors. In this review, we will focus on the recent advances in the pathogenesis of somatotroph tumors.Various genetic and epigenetic factors are involved in the pathogenesis of somatotroph tumors. Although GNAS are the most prevalent cause of somatotroph tumors, germline mutations in various genes  are also known as the cause of somatotroph tumors. Moreover, recent findings based on multiple perspectives of the pangenomic approach including genome, transcriptome, and methylome analyses, histological characterization, genomic instability, and possible involvement of miRNAs have gradually unveiled the whole landscape of the underlying mechanisms of somatotroph tumors. In this review, we will focus on the recent advances in genetic and epigenetic pathogenesis of somatotroph tumors.Acromegaly is caused by excessive secretion of GH and IGF-I mostly from somatotroph tumors. Various genetic and epigenetic factors are involved in the pathogenesis of somatotroph tumors. While somatic mutations of  Excessive secretion of GH and IGF-I prior to closure of the epiphyseal line results in giantism with marked increases in height, while increased secretion of these hormones post-closure results in acromegaly. Both gigantism and acromegaly are relatively rare, occurring in only 8 and 11 cases per million people per year, respectively ,3. The 2Densely granulated somatotroph tumors are usually diagnosed at a younger age and when they are smaller than sparsely granulated somatotroph tumors due to being highly hormonally active . DenselyGuanine nucleotide activating subunit (GNAS) gene and the additional portion can be explained by the handful of classified rare germline mutations ..GNAS mutGNAS mutation over the last 20 years. These evaluations have also facilitated the evaluation of various pathological mechanisms in these tumors, revealing that copy number variations and epigenetic abnormalities are also critical components in somatotroph tumors development. However, the cause of the majority of cases with somatotroph tumors remains unelucidated, highlighting the need for additional breakthrough studies. Recent advances in therapeutic strategies have enabled “a targeted therapy” based on the underlying mechanisms and responsible molecules. Therefore, it is critical to clarify the remaining causes to develop tailor-made therapies.Advances in genome analysis methods have led to the identification of several additional novel causative genes to


The Research Topic “Frailty: Risks and management” aims to investigate the recent advances in the risk assessment and management of frailty in older adults, a growing major public health burden due to population aging, through relevant articles proposed by research groups from different countries.The selected articles address the typical problems associated with this geriatric multifactorial condition from different perspectives and points of view, highlighting emerging epidemiological aspects alongside consolidated ones, analyzing risk factors specific to sex, reporting the potential role of new diagnostic tools and helping to propose the multidisciplinary approach as an essential resource in the care of elderly people.Vargas-Torres-Young et al. found that cognitive frailty and its specific components (cognitive impairment and modified Fried Phenotype criteria) were associated with higher risk of mortality, stimulating discussion on the role of interventions aimed at reversing this condition.Cognitive decline is extremely common among older adults, and cognitive frailty represents an emerging nosological entity that selectively associates it with frailty, as a potentially reversible syndrome . FocusinCoelho-Júnior and Uchida investigated the effects of resistance training programs on frailty status, physical performance, cognitive function and blood pressure in 60 Brazilian pre-frail and frail older adults, randomly allocated to low-speed and high-speed exercises. The results are particularly interesting because, although both resistance trainings reversed frailty status and enhanced physical performance, different patterns of improvement were observed between frailty degrees, with effects probably mediated by the heterogeneity of the aging process.Interventions aimed at treating frailty are the subject of intense debate, given that their effectiveness is conditioned by multiple factors. At this regard, Lv et al. enrolled 150 people aged ≥65 years from the First Hospital Affiliated to Nanjing Medical University who had undergone the anterior ultrasound of ulnar, vastus lateralis and anterior tibial muscles. The authors demonstrated that frailty phenotype (Fried's model)   was closty Index  and ultrArantes et al. propose that heart rate variability can constitute a potential marker of frailty, as epiphenomenon of changes in cardiac autonomic modulation. They provide an overview on the tools to monitor the heart rate variability and summarize the evidence on its association with frailty.In recent years, several pieces of literature have been focusing on the variability patterns of measurable variables as measures of the altered state of homeostatic mechanisms underlying the development of frailty, especially in cardiovascular medicine . In thisWang, Lv, et al., based on the results from 13,859 participants in the Chinese Longitudinal Healthy Longevity Study (CLHLS), reported a high prevalence of pre-frail and frail participants  and provided a comprehensive insight into the epidemiology of this syndrome and related adverse outcomes. In their second article  included in the present collection, a corresponding paper on the epidemiology of frailty, the authors presented a wide range of sex-specific contributors to frailty. Indeed, focusing on a group of 3,327 participants from the CLHLS, they reported risk factors that were common for both sexes and others more associated with the male or female sex. The protective effect of greater household income, higher level of physical activity and fresh fruit and vegetable consumption was shown for both sexes.In the era of personalized medicine, with a tailored preventive and therapeutic approach to older adults, studies on epidemiology that take into account the specific risk factors for frailty in individual countries and cultures are needed. Huang et al. found in their 2-year follow-up project that undertaking frequent intellectual activities  corresponded with decreased risk of frailty syndrome in older adults aged 60 years and more. When considering frailty risk factors from a wider perspective, iatrogenic harms come to the fore. In their prospective cohort study of hospitalized patients treated with intravenous infusions, Cao et al. reported that the risk of frozen shoulder within 1 year of hospital discharge was as high as 5.2%. The risk factors for its onset included longer time of intravenous infusion, longer hospital stay, older age and comorbidities.The two above-mentioned Chinese studies are accurately supplemented by the results of the prospective observational China Health and Retirement Longitudinal Study (CHARLS). Gao et al. on the relation between hunger in childhood and frailty in old age, seems particularly important. In their cross-sectional analysis of data obtained from the 2018 Chinese Longitudinal Healthy Longevity Survey, the authors showed that experience of childhood hunger was linked to frailty in late life, mediated by age and financial resources.The current global economic and political crises recall the key role of socio-economic support on the state of health of the various age groups of the population, with a medium-long term impact. In this context the research question of the research proposed by Crooke et al. presented a comprehensive scope of presbyopia as an opportunity for the timely detection of pre-frailty and frailty.Taking advantage of the multidisciplinary approach, which is strongly advocated for the proper management of the complexity of frailty syndrome, geriatricians should go hand in hand with ophthalmologists when working with middle-aged and older patients. The opinion paper included in our collection by Granata et al. propose a systematic review to evaluate the use of Clinical Frailty Scale, a screening tool based on clinical judgment (A key component of the aging process is represented by comorbidities, which, together with chronological age, constitute the main factors associated with frailty. judgment , for fraIn summary, appreciating the high impact of frailty on national health systems, the articles in this Research Topics collection provide a meaningful and up-to-date scenario on some key aspects of this syndrome, also pointing out interesting potential innovations and stimulating new concepts. The Research Topic “Frailty: Risks and management” represents an important contribution to the body of scientific evidence in the field of geriatric medicine and also reveal current research gaps, stimulating ideas for future research on the topic.LB and KP wrote and approved the editorial. All authors contributed to the article and approved the submitted version.


Over past years, the fabrication of Ti-based permanent implants for fracture fixation, joint replacement and bone or tooth substitution, has become a routine task. However, it has been found that some degradation phenomena occurring on the Ti surface limits the life or the efficiency of the artificial constructs. The task of avoiding such adverse effects, to prevent microbial colonization and to accelerate osteointegration, is being faced by a variety of approaches in order to adapt Ti surfaces to the needs of osseous tissues. Among the large set of biocompatible materials proposed as an interface between Ti and the hosting tissue, diamond has been proven to offer bioactive and mechanical properties able to match the specific requirements of osteoblasts. Advances in material science and implant engineering are now enabling us to produce micro- or nano-crystalline diamond coatings on a variety of differently shaped Ti constructs. The aim of this paper is to provide an overview of the research currently ongoing in the field of diamond-coated orthopedic Ti implants and to examine the evolution of the concepts that are accelerating the full transition of such technology from the laboratory to clinical applications. The rapid increase in the aged population is creating an ever increasing demand for the replacement of dysfunctional bone tissues with artificial constructs.Over the past decades, exciting advances in material science have enabled substantial improvements in prosthesis, fracture fixation and replacement components. Beyond efficacy, safety and reliability, the increased comfort of patients is also required. In this context, the intersection of material science and biomedicine is called upon to solve the pressing challenges of designing and producing ceramic, polymeric and metallic osteobiological grafts able to fully substitute for conventional allografts, autografts and xenografts . In partThe wide range of metallic devices designed to restore or replace damaged or diseased hard tissues include orthopedic implants  as well as components for dental restoration . Nowadays, the production of such tools used in bone and dental implantation surgeries far exceeds any other biomaterial applications. As an example, each year in the United States, over 800,000 knees and 350,000 hip replacements are made, with a total of 2,244,587 hip and knee arthroplasty procedures performed between 2012 and 2020. In 2021, a cumulative procedural volume growth of 18.3% occurred compared with the previous year [As regards to metals, the stringent criteria required by load-bearing applications limit the options to the few with mechanical properties similar to natural bone, i.e., stainless steel, cobalt-chromium, and titanium. Titanium (Ti) and its alloys are certainly the synthetic materials more frequently used for permanent implants. Ti is a metal with no allergenic or immunogenic effects and it is characterized by a strength-to-weight ratio that enables a variety of load-bearing applications . MoreoveTo allow integration of the implant into the host tissue, the cell’s metabolism requires a local microenvironment able to assure cellular communication, the transport of oxygen and nutrients, and the removal of wastes produced. These complex regenerative processes at the bone-to-implant interface depend also upon physical and mechanical cues. The cells receive mechanical stimuli from the artificial environment and translate them into biochemical signals. Such signals modulate several aspects of cellular behavior, including attachment, proliferation, and differentiation. The achievement of successful bone remodeling is currently driving research towards the development of innovative implant materials with engineered surfaces that mimic the natural extracellular matrix (ECM) ,7,8.From a biological point of view, even if Ti is not intrinsically an osteoinductive and osteoconductive material, the research of the last two decades have highlighted that effective osseointegration can be induced on Ti surfaces via proper modulation of the chemistry, topography and surface energy ,18,19,20However, decades of implantation surgery have provided evidence that the body-fluids induce in Ti several adverse processes, such as wear, corrosion, and eventually, tribocorrosion .These processes are particularly strong when Ti components are inserted into rather aggressive environments, such as the oral one ,23,24. Therefore, surface engineering of the Ti-based implants and the improvement of the interface between the implants and the host tissue have become a key task ,14.A great variety of mechanical, physical and chemical treatments have been proposed to modify the Ti surfaces and to control the finishing of the prostheses. These strategies range from sand-blasting using abrasive particles to etching induced by acids, high voltages or plasmas ,27,28,29Nevertheless, it has been found that osteopromotion is not the only biological process taking advantage of the micro and nano features present on modified Ti surfaces. In effect, the rough surfaces create an environment that is also favorable to bacterial adherence and biofilm formation ,31,32. IOver the last two decades, a commonly employed strategy to prevent infections of periprosthetic joints has been the coating of Ti biotic surfaces with antimicrobial layers . To thisHowever, several disadvantages have been highlighted in the various approaches aimed at preventing the spread of bacteria and biofilm formation. Among the more severe drawbacks, there are the adverse effects of antibiotics on tissue integration, development of antibiotic resistance, and cell damage caused by the nonantibiotic organic antimicrobial agents ,40. MoreThe review of literature reveals that, in general, the enhancement of bone mineralization and the control of bacteria colonization, the two main factors that assure a good integration of implants, have been addressed separately . This isFrom this perspective, ongoing research concerning the design of long-lasting implantable constructs is now focusing not only on simultaneous osseointegration improvement and biofouling contrast, but also on complying with the mechanical properties required for prolonged and intense use .The latest advances in the engineering of orthopedic Ti-coatings are being provided by the use of diamond, a material characterized by chemical stability, high hardness, corrosion resistance, antiwear properties, biocompatibility and cytocompatible behavior ,50,51.Due to their fascinating physicochemical properties, diamond-based materials have been progressively applied in several biomedical areas, from drug delivery to cellular imaging, and from labelling to assembling of multifunctional scaffolds for tissue engineering . Regarding this latter application, from the beginning of the 2000s it has been evidenced that diamond surfaces enhance and, above all, selectively control the osteoblast-like MG 63 cell growth ,54,55,563-coordinated carbon, either in the form of polycrystalline diamond films or micro/ nano-sized diamond grains, behaves simultaneously as a promoter of osseointegration and as an antibacterial agent [Studies on the colonization of artificial diamond surfaces by a series of cells, such as fibroblasts, osteoblasts and mesenchymal stem cells had evidenced that spal agent ,59,60,61al agent . Even ifal agent , surfaceal agent . It is tal agent .Moreover, to favor the bioactivity of the cells and to promote subsequent tissue integration, the mechanical properties of a material must match those of the tissue to which it is applied . This reInvestigations performed using different cell lines and substrates with various degrees of stiffness have demonstrated the dependence of cell morphology, motility, proliferation and differentiation on the elastic modulus of the substrate and on its adjustments ,65,66,67The good compatibility of diamonds with MG 63 cells can be rationalized by considering that the mechanical properties of artificial diamond layers, even if dependent on their structure ,69, matcTaken together, all of the studies strongly highlighted the potentialities of diamond as a favorable material for bone tissue–implant interfaces and evidenced that the cellular response to diamond is influenced by a complex interplay of texture, microstructure, surface chemistry and surface potential . As regards the bioactive behavior of the inserted foreign materials, one must consider the differences among the chemical-physical-biological environments surrounding the various implants ,72. DentThe new scenario opened by diamond has led us to explore all of the options available to realize diamond–titanium systems. The engineering of Ti-diamond components can be accomplished by a variety of methods, from the synthesis of crystalline coatings using CVD-based techniques to the embedding of diamond nanopowders onto the surfaces of Ti implants.A widely adopted technique for Ti coating by diamond is the Chemical Vapor Deposition (CVD), which produces a diamond phase using gaseous mixtures of carbon-rich reactants and hydrogen activated by hot-filaments or Micro-Wave (MW) plasmas . The CVDModulation of the layers’ texture and morphology can be achieved by varying the composition and flow rate of the gaseous phase. In particular, the addition of an inert gas during the CVD process increases the diamond nuclei density and enhances secondary nucleation, promoting the growth of nanosized grains . The morThe deposition of crystalline diamond layers can be accomplished also on non-planar substrates. A mandibular movement simulator has been employed to perform functionality tests on diamond-coated Ti components that imitate condyle–fossa pairs. As reported in , the coaThe CVD technique also enables nanocrystalline diamond to be coated onto fiber-shaped substrates, as SiOx micro-fibers . Mats ofAs a whole, the use of the CVD technique proved to be very effective for diamond coatings in implantology. This process of diamond deposition is, indeed, both resource-efficient and cost-efficient and enables a series of differently shaped substrates to be coated .The nanodiamond produced by detonation processes, commonly referred to as “detonation nanodiamond” (DND), was first explored in the 1980s . As repoMoreover the detonation nanodiamond is characterized by its ability to create a tunable surface through a number of easy functionalization processes ,85. As aThe coupling of DND with Ti is mainly achieved by inserting the powders into polymeric matrices and by using such composite materials for the coating of implants ,87. ThinThe additive manufacturing processes, that make it possible to create custom-made products by means of computer assisted technologies, are overcoming the limitations of standard techniques and are now finding wide applications in the field of implantology . A detailed description of the additive manufacturing processes used for fabrication of Ti bone implants, along with some outstanding case studies, is given by Popov et al. . In partDirect deposition and integration of nanodiamonds  or of poBeyond their intrinsic extraordinary properties, diamond coatings can be further morphologically and chemically modified in order to best address the specific demands of biological tissues.There are several ways to induce structural changes in the diamond surfaces and to produce substrates with a topography mimicking the main features of the cell microenvironment. Among a variety of settled techniques, the most efficient are molten salts etching , solid-pPlasma treatments represent a powerful strategy for modulating the topography of diamond layers. Arrays of vertically aligned nanowhiskers can be obtained in plasma-reactors by the H-etching of self-assembled detonation nanodiamonds . As showConversely, also surface smoothing can be achieved by proper plasma processes. As described in , the tre2-groups [Plasma processes are also able to modify the surface chemistry of the diamond layers. Indeed, it is possible to saturate the dangling bonds of diamond with H and O atoms, or with NH2-groups .It is interesting to note that modifying the surface chemistry induces changes in the value of the surface free energy and, therefore, heavily influences the biological response of the material. A series of in vitro studies evidenced strong differences in the behavior of cells contacting micropatterned diamond surfaces with specific chemical terminations.3-plasmas, for which a high free energy (62.9 mN m−1) was measured; and the worst occurred on H-treated surfaces with lower surface free energy (40.2 mN m−1). This study established a monotonic correlation between surface free energy and the wettability of diamond and validated the dependence of the osteogenic potentialities of diamond on its surface functionalization.A strong correlation between the surface free energy of diamond and osteoblast responses was evidenced in 2012 by Yang et al. . In thisAfter that, several other researches have been focused on the task of modulating the wettability of the coatings, developing plasma treatments able to generate diamond surfaces with either hydrophobic or hydrophilic characters, and tailoring the level of such properties . Figure More recently, Broz et al. , using r2-termined hydrophilic nanocrystalline diamonds for osseointegration and bone formation [Proteome studies of human osteoblast MG63 cells confirmed the potential of O- and NHormation ,107. Theormation . In addition, Fong et al. , in theiHydrophilicity is presently considered a key issue in implant engineering because it is related not only to high biocompatibility and to antibacterial effects, but also to a lower friction coefficient. This last factor, associated with wear behavior of the material, is important, especially for joint arthroplasty.Beyond the topographical features and the surface chemistry, conductivity and surface charge also influence the cell–substrate interactions, mediating the initial plasma protein binding and impacting the whole growth process ,108.Surface conduction has long been recognized as a fundamental requirement for the adhesion and growth of cardiac and nervous cells . HoweverTherefore, a fundamental task in both regenerative medicine and implantation surgery is the modification of the diamond’s insulating characteristics. In the field of implantology, charged surfaces offer the additional benefit of strongly reducing bacterial adhesion . Electrical conducting n-type diamond layers are obtained by default when the CVD-growth is carried out using a gaseous phase rich in Nitrogen (N), but a proper modulation of conductivity is commonly achieved by inserting Boron (B) inside the diamond lattice. The B-doping, which induces a p-type semiconductivity, is addressed by injecting B-containing gaseous compounds during the deposition process. Several studies have confirmed the capabilities of B- and N-doped diamonds to facilitate cellular adhesion and to contrast biofilm formation ,113,114.The possibility of inducing further functionalities, such as electrical conductivity, by adding a variety of foreign species to the diamond phase is an attractive side to CVD methodology . To overThese findings are in line with those obtained by Fox et al. , who proNo other field of material science faces such a diverse set of challenges as the one of biomedical materials. A look at the huge number of recent papers dealing with orthopedic and dental implantology evidences a rapid evolution of this research area, with a trend following the advances in regenerative medicine and material engineering.In a living organism, materials specifically designed to match the requirements of the biological environment can also be affected by the physicochemical reactions of tissues against a foreign body and can undergo severe degradation. This also applies to Ti, the leading material used in orthopedic implantology, which still had to face such difficulties from the beginning of its use in clinical practice. Among the Ti surface modifications proposed to prevent, or at least mitigate, adverse reactions, the most interesting could be the coating of Ti by the biocompatible and chemically inert diamond.Diamond coating offers inherent protection against the common degradation phenomena induced in Ti by body fluids. However, it has been discovered that the chemical protection of Ti provided by the diamond coating does not automatically guarantee favorable outcomes. In effect, the diamond-coated orthopedic and dental replacements produced first resulted in poor osseointegration and infection occurrence.The disappointing performance of the first diamond-coated Ti implants in promoting bone regeneration were rationalized when the progression of research highlighted that stiffness, micro or nano-roughness, and surface texture of the diamond could no longer be considered the only parameters triggering the attachment and proliferation of osteoblasts. Conversely, a deeper understanding of intercellular biochemical communications evidenced how a lot of additional features, including surface texture, topography and microstructure, surface chemistry, surface free energy and electroconductivity, contribute to the role of diamond as viable osteoblast–Ti interface. These indications pointed towards the need for precise engineering of the diamond surfaces to enhance osteointegration and reduce bacterial growth. However, looking through the literature, one can notice that, in general, the issues of osseointegration and that of biofouling mitigation have been addressed separately. Only in the last few years have proposed technological solutions been able to face these problems in a combined way. As briefly summarized in this review, a variety of chemical and physical approaches are being designed, nowadays, to adapt the features of the diamond coatings to use in Ti permanent implants. The control of the morphology, topography and surface texture of the diamond coatings is still an important task, which can be accomplished by varying the composition of the reactant vapor phase during the CVD processes or by performing specific pre-treatments of the Ti surfaces. In addition, the use of detonation nanodiamonds as the starting material enables the morphological features of the coatings to be controlled, either when the nanodiamond powders are inserted into the polymeric matrices or when a dip-coating methodology is used. Moreover, the controlled modification of diamond surfaces can be obtained via post-process protocols. Among the variety of chemical reactions developed to produce the surfaces and replicating, at best, the features of the extracellular matrix, plasma processing is certainly the leading technique for the micro- and nano-shaping of diamond layers. A further advantage of this technique is that proper plasma treatments make it possible to manipulate, beyond the topography, also the chemistry and therefore, the free surface energy of the coatings. This is a key point, because it is the value of this last parameter that manages the wettability and determines whether the applied coating will promote the attachment of cells to the implant. 2-) is a deeply pursued task, because the effects of the hydrophilicity and hydrophobicity of materials on osteoblast adhesion and integration have become increasingly evident. Rather, it is now confirmed that oxidized diamond surfaces with high hydrophilicity are strongly requested in implantology, not only for improving cellular response, but also for counteracting biofouling and for reducing friction between the contacting surfaces.Nowadays, the generation of a surface terminated by specific atoms (mainly H and O) or groups  and non load-bearing  implants.The combination of so many specificities requires the use of manufacturing approaches able to offer great flexibility in producing, engineering, and processing diamond coatings. It is evident that such a task has no single solution, and that any attempt to meet the challenging issues of osteogenesis, bacterial adhesion, and cell viability at the diamond-coated surfaces of implantable devices needs to put into play all of the techniques and protocols settled up until now and to envisage other hitherto unexplored approaches.


Aspergillus section Flavi. Aflatoxin contamination of food, feed, and agricultural commodities is a serious economic and public health concern not only due to hepatotoxic, teratogenic and carcinogenic effects of this toxin, but also for food spoilage and deterioration result from the growth of producing fungi on susceptible substrates. At present, aflatoxin contamination is a real challenge for developing countries with increasing reports of involvement from new parts of the world due to global warming problem. With expanding our knowledge about detrimental effects of aflatoxins on public health and economy, new technologies are rapidly developed to uncover novel aspects of these dangerous mycotoxins from the taxonomy, biochemistry, and evolution of producing fungi to effective strategies for pre- and post-harvest management of aflatoxin contamination. After a successful endeavor on aflatoxins in volume I , this Research Topic as volume II has highlighted recent advances in distribution and management of aflatoxins and aflatoxin-producing fungi in food, food products and agricultural commodities in relation to their impact on public health, economy and agriculture. Within this topic, five articles have been published that complemented our knowledge of the occurrence of aflatoxins, regulatory rules, diversity, and biocontrol of aflatoxin-producing fungi.Food safety issues have received major consideration not only for increasing numbers of natural and synthetic contaminants but also in relation to ever increasing world population and subsequent demand for food and food supplies. Mycotoxins produced by filamentous fungi are considered as hazardous food and feed contaminants with global distribution. Aflatoxins, the best-known mycotoxins ever, are secondary toxic metabolites produced by diverse groups of filamentous fungi mainly belonging to the Atehnkeng et al. evaluated the efficacy of “Aflasafe” as a biocontrol product in sub-Saharan Africa, where the agricultural crops especially maize are susceptible to aflatoxin contamination. Aflasafe is composed of a cocktail of atoxigenic Aspergillus flavus genotypes native to the region to mitigate aflatoxin contamination. The authors examined Aflasafe in nine maize fields for 3 years, under various conditions to evaluate its biocontrol effectiveness. After quantitation of aflatoxins in grains at harvest and after simulated poor storage, it was shown that biocontrol ability and the genotype viability decreased in a condition which did not support annual treatment. The authors showed that aflatoxin content of maize treated with Aflasafe up to 3 years reduced meaningfully in grains at harvest in comparison with untreated maize. It is interesting to point out that the genotypes were able to survive in the soil in a manner that efficiently reduces maize contamination for 3 years. In conclusion, the authors indicated that although efficient reduction in aflatoxin concentration requires annual applications of Aflasafe, carry-over established in their study can reduce both application frequencies and associated costs.In inclusive research on the reduction of aflatoxin contamination in maize, Ahmad et al. expanded the current knowledge about using natural inhibitors as a sustainable way to combat the spreading of aflatoxigenic fungi and contamination with aflatoxins. Since physical and chemical pre- and post-harvest control strategies of aflatoxin management are laborious and lead to increase in toxic fungicide residues on food and the environment to various extents, the authors discussed various practical and potential application of plants, algae and microorganisms including fungi and bacteria in growth inhibition of aflatoxigenic fungi and subsequent aflatoxin contamination. The role and mode of action of phenolic compounds, antioxidants, plant essential oils, and aromatic compounds such as terpenoids and flavonoids, mushrooms and micro-fungi metabolites especially those obtained from Lentinula edodes and Trametes versicolor, and finally antimicrobial peptides in inhibiting fungal growth and subsequent aflatoxin contamination was discussed. These natural substances either inhibit the growth of these toxigenic fungi and aflatoxin contamination in food, feeds, and food supplies or suppress aflatoxin biosynthesis and removing or degrading aflatoxins. The authors concluded that extensive studies are still needed to fully comprehend the mode of actions of these natural inhibitors at cellular and molecular levels via profiling of transcripts, proteins, and metabolites before they will be applicable for bulk production for industry application.In a comprehensive review on aflatoxin inhibitors, Singh et al. used chitosan nanoemulsion (CsNe) to encapsulate Aniba rosaeodora essential oil (AREO-CsNe). Using spectrometry analyses, the authors showed that AREO was successfully encapsulated into CsNe and control released in sufficient amounts. AREO-CsNe was shown to be able to completely inhibit A. flavus growth and AFB1 more efficiently than that of AREO alone. Regarding the AREO-CsNe mechanism of action, it is believed that the fungal plasma membrane is the main target due to impaired ergosterol biosynthesis and enhancement of fungal cell leakage in AREO-CsNe exposed fungus. In silico molecular docking studies also showed efficient interaction of linalool, the main component of A. rosaeodora essential oil with Ver-1 and Omt-A proteins, which are known as essential enzymes in AFB1 biosynthetic pathway. AREO-CsNe showed enhanced antioxidant activity and in a food model system of stored millets by using Setaria italica seeds, induced complete protection against both lipid peroxidation and AFB1 contamination in a 1 year period. In conclusion, the authors indicated that enhanced activity of AREO-CsNe over AREO makes it a promising candidate to preserve stored millets and possibly other foods or grains from fungal growth and AFB1 contamination.In a promising study on developing of a chitosan-based natural nanoemulsions as improved antifungal and anti-aflatoxigenic agents, Yunus et al. evaluated the importance of knowledge of Farmers and potential ways to control aflatoxin contamination of raw milk in Pakistan. The authors showed that while the majority of the farmers were familiar with the negative impression of fungal-contaminated feed on animal health, <50% of them were conscious of the possibility of transferring aflatoxins from contaminated feed to milk. They concluded that availability of certified aflatoxin-safe feedstuffs, subsidy on quality feeds for animal nutrition, raising Farmer's awareness and legislation and monitoring could effectively impact the fate of aflatoxin contamination of milk.Contamination of milk and other dairy products with aflatoxin is another major concern and public health problem. As a conclusion, data from the present Research Topic highlights the necessity of developing novel strategies for pre- and post-harvest control of aflatoxigenic fungi and aflatoxin contamination of food, feed, and agricultural commodities and expanded our current knowledge about the importance of fungal natural inhibitors and their novel improved formulations in aflatoxin control and prevention strategies.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


Here, we mainly focus on the construction of brain organoids and review the application of brain organoids in disease modelingand drug screening.Brain organoids are produced by the differentiation of pluripotent stem cells under three-dimensional culture conditions by adding neurodevelopment-related regulatory signals. They are similar to the cell composition and anatomical structure of the brain, and can reflect the developmental process of the brain, as well as their physiology, pathology, and pharmacology. Brain organoids are good models to study human brain development and brain-related diseases  Compared with traditional cell models, organoid models have more stable genetic and phenotypic characteristics during in vitro culture and more abundant cell types  cell culture product, which is an organ-specific product obtained by inoculating stem cells in matrigel or basement membrane extract under the action of a specific cytokine mixture . Organoill types .etc.The brain is the largest and most complex organ with complex neural activity . The powThe development of stem cell technology enables researchers to use human-induced pluripotent stem cells (hiPSC) to induce brain-like tissues and organs from a 3D perspective . Brain oNeurodevelopmental disease (NDD) refers to the abnormal development of the nervous system caused by hereditary or acquired etiology, resulting in brain dysfunction, including intellectual disability, autism spectrum disorder, attention deficit hyperactivity disorder, and other diseases. Analyzing the pathogenic mechanism of neurodevelopmental diseases has always been one of the key topics in neurobiology. But due to the ethics restrain, human and non-human primate brain tissue shortage, even though a small number of patient tissues can be obtained, it can only reflect the disease in the terminal stage, the occurrence and development of the disease cannot be analyzed. Therefore, the knowledge and understanding of human brain developmental diseases are mainly derived from studies on rodents. However, the complex structural and functional partitions unique to the human brain cannot be fully reproduced by animal models. The development of stem cell technology, especially the establishment of induced pluripotent stem cells, provides an ideal human cell model for decoding the pathogenesis of neurodevelopmental diseases. Reprogramming of patient somatic cells with disease genes into induced pluripotent stem cells (iPSCs) followed by differentiation into various types of nerve cells has been widely used in the study of various neurological diseases. However, neurological disease phenotypes are highly heterogeneous, including abnormalities in brain structure such as brain size, and problems with synaptic activity. For this complexity, traditional two-dimensional (2D) neural culture can only provide limited insights. Subsequently, a 3D brain organoid culture protocol was pioneered by improving the 2D neural culture method, opening new horizons for the study of neurodevelopmental diseases.in vivo microenvironment, and is difficult to reproduce the complex phenotypes of neurological diseases, but also breaks through the lack of human-specific genetic characteristics, brain regions, and functions, difficulties in comprehensively simulating the development of the human brain and the limitations of disease occurrence and development. It is an important tool to study human brain development and evolution in vitro, to explore the interaction of different brain regions, brain, and other organs, and carry out disease simulation and drug screening in vitro. This article will review the history of the establishment and development of brain organoids, introduce the progress of brain organoids in the exploration of brain development, neurological disease simulation, and drug screening, and analyze possible future directions.The generation of brain organoid technology not only makes up for the shortcomings of traditional 2D cell culture that cannot simulate the complex structure of brain tissue and the in vitro model used for brain research was the embryoid body (EB)-derived neural rosettes (NR). Neural rosette is a human neural tube-like structure that appears during the differentiation of hiPSCs into cerebral organoids and is the basis for the differentiation of various nerve cells. The further development of hiPSCs technology has promoted the generation of induced pluripotent stem cell neurospheres and three-dimensional neuroepithelial tissue, which can reflect the genetic characteristics of the human brain to a certain extent, but they do not form a complete and complex brain structure, and they lack the coordination between different sub-subsections of the brain.In 1981, researchers isolated embryonic stem cells from the inner cell mass of mouse blastocysts . In 1998However, traditional neuronal diseases are simulated using neurons differentiated and developed from human pluripotent stem cells (hPSCs), which is also a 2D culture-based system . Althougin vitro, and can better maintain human-specific genotypes and protein expression levels. With the advancement of bioengineering technology, the development of pluripotent stem cell-derived 3D brain organoid technology has become more mature, and its application prospects have become broader and broader. Taken together, pluripotent stem cell-derived 3D brain organoid technology can be used to break through the current bottleneck in the study of brain development and neurological diseases and has great potential as a model for brain development and neurological disease research.In 2013, Current research on neurological diseases mainly uses animal models. However, the genetic background, and physiological and pathological characteristics of model animals are different from those of humans. So it is difficult to simulate the occurrence and development of diseases in human brain truly and comprehensively. As an effective complement, human brain organoids provide new tools for modeling human brain development, psychiatry, and degenerative diseases, as well as for drug screening and gene therapy.etc. Another group found that the volume of brain organoids with RAB39B mutation increased, and the excessive proliferation of neural progenitor cells led to the thickening of the SOX2 positive ventricular zone (VZ), leading to differentiation disorder. Also, the lack of RAB39B causes the PI3K-AKT-mTOR signaling pathway excessively activated. And inhibiting the PI3K-AKT-mTOR signaling pathway can rescue the autism phenotype (Autism spectrum disorder (ASD) is a brain developmental disorder characterized by language impairment, social difficulties, and repetitive stereotyped behaviors . Marianihenotype .in vivo  deposition and neurofibrillary tangles  . Howeverin vivo . Anotherin vivo . This syrganoids . In addiin vitro culture models, whether monolayer culture or tumor sphere culture, may take a lot of time to establish and use exogenous epidermal growth factor, basic fibroblast growth factor, and/or serum to serially pass tumor cells in a clonal expansion fashion, which is not conducive to maintaining the various cell subtypes and key driver gene expression of the parent tumor, and lack of organoid histological features and interaction between tumor and normal tissue  is the most common and aggressive primary malignant brain tumor . Despitel tissue . An altel tissue . In addiin vitro, including brain organoids  and hexosaminidase B (HexB) activity . At the in vitro by 2D neural culture and 3D brain organoids, respectively (Fragile X syndrome (FXS) is an intellectual disability syndrome caused by mutations in the FMRP gene on the X chromosome . Researcectively . CompareLissencephaly, also known as the smooth brain, is a neuronal migration disorder . Among tAutosomal recessive primary microcephaly is a neurodevelopmental disorder caused by autosomal aberrations, which is mainly characterized by small brains, especially cerebral cortical changes . Among tin vitro model, have played an important role in the study of psychiatric disorders. Schizophrenia is a highly heritable mental disorder, and a study based on DNA whole-genome sequencing of schizophrenia patients found that 15q11.2 gene copy number variation is one of its risk factors (Brain organoids, as an emerging  factors . Yoon et factors . Other g factors .Brain organoids are expected to become an efficient platform for drug development, screening, and testing. Drug safety evaluation is an important reference for drugs to enter the clinical stage. Brain organoids are expected to become important models for drug screening and reduce the testing burden of clinical trials due to their high molecular and structural similarity to the source tissue. Zika virus research with brain organoid technology not only reveals the relationship between Zika virus outbreaks and the incidence of congenital microcephaly but also has great potential for drug testing, including the preparation of potential Zika virus antiviral drugs. In addition, brain organoids are also widely used in other common drug screenings. Some studies have used a brain organoid model constructed by neural progenitor cells, glial cells, and neurons to explore the effects of METH on the human brain . The stuin vitro organoid culture technology has experienced rapid development and has shown strong application value. In general, organoids can well mimic the corresponding tissues in patients at the gene level and morphological characteristics, are also suitable for high-throughput drug screening and provide a research model for personalized treatment of diseases.Since its establishment, in vitro, cells in the middle of the organoid undergo massive apoptosis due to a lack of oxygen and nutrients. Therefore, the establishment of an improved circulatory system of brain organoids is the general trend. In addition, current brain organoid technologies still suffer from significant limitations, especially in the functional assessment of neural network activity and cellular interactions, and neural circuit functions relevant to neurodevelopmental and neuropsychiatric pathologies (Human brain organoids have attracted great interest in recent years due to their potential to study human brain development and neuropathology without the constraints of animal models. The development of 3D brain organoid technology is less than 10 years and is still in its infancy. In terms of their cellular and molecular composition, the current architecture of brain organoids can mimic the second-trimester human fetal brain. However, since cerebral organoids lack vascular circulatory system, they mainly rely on free diffusion for oxygen exchange and nutrient uptake from the culture medium. When cultured for a long time hologies .In summary, although the current brain organoid culture system still has technical defects, it does not have the 3D structure and complex functions of human natural organs. However, it is undeniable that the 3D brain organoid model has brought great progress to human research on brain development and disease mechanisms . The con


Lysosomes are organelles containing acidic hydrolases that are responsible for lysosomal degradation and the maintenance of cellular homeostasis. They play an important role in autophagy, as well as in various cell death pathways, such as lysosomal and apoptotic death. Various agents, including drugs, can induce lysosomal membrane permeability, resulting in the translocation of acidic hydrolases into the cytoplasm, which promotes lysosomal-mediated death. This type of death may be of great importance in anti-cancer therapy, as both cancer cells with disturbed pathways leading to apoptosis and drug-resistant cells can undergo it. Important compounds that damage the lysosomal membrane include lysosomotropic compounds, antihistamines, immunosuppressants, DNA-damaging drugs, chemotherapeutics, photosensitizers and various plant compounds. An interesting approach in the treatment of cancer and the search for ways to overcome the chemoresistance of cancer cells may also be combining lysosomotropic compounds with targeted modulators of autophagy to induce cell death. These compounds may be an alternative in oncological treatment, and lysosomes may become a promising therapeutic target for many diseases, including cancer. Understanding the functional relationships between autophagy and apoptosis and the possibilities of their regulation, both in relation to normal and cancer cells, can be used to develop new and more effective anticancer therapies. Neoplastic diseases, despite significant advances in treatment methods, are still the second most common cause of death, right behind cardiovascular diseases. One of the main reasons for the failure of anticancer therapy is the phenomenon of multidrug resistance to the cytostatics used, often occurring in cancer cells as a result of various interactions . The proThus, the task of modern oncology is to search for new strategies to overcome the resistance of cancer cells to the therapies used, as well as to understand the mechanisms of this phenomenon.Lysosomal cell death, which occurs with the participation of the lysosomal system, may play an important role in anticancer therapy. The main reason for drawing attention to this type of cell death is the specific structure and role of lysosomes in both normal and malignant transformed cells, which makes lysosomes a unique target for the action of anticancer drugs. This work focuses on the mechanism of lysosomal death, based on the destabilization of the lysosomal membrane and on pharmacologically interesting compounds whose potential can be used to induce lysosomal membrane permeability. Compared to other types of cell death, such as apoptosis, lysosomal cell death can be very useful in eliminating the phenomenon of multidrug resistance, which is one of the main causes of failure when using anticancer therapies.+-ATPase family, which ensures the acidic pH inside the lysosome . Th. Th70]. As a consequence of the increased permeability of the lysosomal membrane in cancer cells, the death pathway may occur, which runs independently of executive caspases, during which cathepsins leak and cytosol acidification and caspase-2 activation occur .However, it has been shown that cancer cells can inhibit both the apoptotic and lysosomal pathways in order to differentiate and proliferate . An incrUnder conditions of cellular homeostasis, the degradation of molecules, cell organelles and cell fragments occur by macroautophagy involving the fusion of primary lysosomes containing a set of lysosomal enzymes (hydrolases) with autophagic vacuoles (autophagosomes). In contrast, sequestration of molecules or cell fragments into lysosomes occurs by microautophagy.LMP inducers cause a slight leakage of cysteine proteases (cathepsins) into the cytoplasm, which activate pathways leading to cell death. Complete disruption of the lysosomal membranes leads to a massive release of lysosome components into the cytoplasm, causing necrosis. LMP-inducing agents include lysosomotropic compounds, some antihistamines, immunosuppressants, DNA-damaging drugs, chemotherapeutics, photosensitizers, plant compounds . List of abbreviations: LMP—lysosomal membrane permeability, ROS—reactive oxygen species, MOMP—mitochondria outer membrane permeabilization.The available literature describes a number of agents that cause modulation of the lysosomal compartment, an increase in lysosomal enzyme activity (including cathepsins) and an increase in the lysosomal membrane permeability (LMP) including its damage, which can lead to the release of lysosomal proteases and their proapoptotic effect ; Table 1Lysosomotropic compounds have a high affinity for accumulating in lysosomes (lysosomotropism), and many of them have therapeutic activity (cationic amphiphilic drugs—CAD) . The CADDespite belonging to different pharmacological classes, the aforementioned compounds share the same physicochemical properties. Common to all CADs is their amphiphilic nature, which is due to their hydrophobic ring structure and hydrophilic side chain containing cationic amino groups . AccordiAn interesting group of substances are lysosomotropic detergents (LDs), which combine features of lysosomotropism with detergent activity . These aAn example of endogenous lysosomotropic agents is sphingosine, generated in the lysosomal membrane by the sequential action of acid sphingomyelinase (aSMase) and acid ceramidase, which convert sphingomyelin to ceramide and then to sphingosine . SphingoDamage to the lysosomal membrane may also result from the inhibition of acid sphingomyelinase (ASM), which, as demonstrated by Petersen’s team in 2013 , moves iThe strategy of targeting lysosomes by siramesine described in Dielschneider’s work may also be a new form of treatment for chronic lymphocytic leukemia (CLL cells). Literature data  show thaAnother group of compounds that affect the lysosomal compartment are some immunosuppressant drugs. One of these is FTY720, a sphingosine-1-phosphate (S1P) analog, which is used to treat multiple sclerosis. FTY720 has also shown anti-tumor activity, but the molecular mechanisms are not fully clear. A study by Min and Kwon in 2020 on glioma cells  showed FTY720 accumulating in the lysosome-induced permeabilization of the lysosomal membrane. On the other hand, the inhibition of LMP by HSP70 and the cathepsin inhibitors blocked FTY720-induced cell death, suggesting that this compound induces glioblastoma cell death after permeabilization of the lysosomal membrane .Research on the assessment of the potential anticancer mechanisms of antihistamines is part of the current canon in the fight against cancer, which consists in the recession of already approved drugs with proven safety and which translates into shortening the time to obtain a drug for oncological treatment. Lysosomotropism is a feature of some representatives of antihistamines, characterized by different physicochemical properties related to their cationic-amphiphilic nature (CAD antihistamines) .According to the study, astemizole, loratadine and ebastine have a high volume of distribution, which translates into the efficient distribution of the drugs into tissues, and because they are weak bases, they accumulate more effectively in acidic tumors than in healthy tissues with a neutral pH. The study authors also suggest that the use of the aforementioned CAD antihistamines in combination with chemotherapeutics can sensitize cells to chemotherapy and reverse multidrug resistance, thereby effectively enhancing the anti-tumor response. Therefore, it is important to expand studies to evaluate other representatives of the antihistamine group of drugs. In our study , we showThe research of Richardson’s team shows that the lysosomal pathway can also be selectively activated by metal chelating compounds from the thiosemicarbazone di-2-pyridylketones (DpT) group, which are characterized by a strong and selective anticancer effect . The afoCompounds that are used in cancer phototherapy also play an important role in lysosomal cell death; an example is N-aspartylchlorin , a photosensitizer with a very high affinity for cancer cells and which accumulates in lysosomes and contributes to the generation of singlet oxygen. The reactive oxygen species generated by the photodynamic process cause rapid damage to lysosomes, leading to the rapid release of cathepsins into the cytosol, the activation of the Bax protein that initiates the mitochondrial apoptosis pathway, and procaspases 3 and 9 ,135. A sCatharanthus roseus alkaloids) [Taxus brevifolia [Compounds that induce LMP include plant antimitotic drugs, i.e., vinblastine, vincristine, vinorelbine  ,136, pacevifolia ,91,137, evifolia , and somevifolia ,138, genevifolia  and azitevifolia .Factors damaging the lysosomal membrane are also other compounds of plant origin, such as resveratrol  and its Aloe barbadensis Mill., Aloe arborescens Mill.), as well as from Frangula alnus Mill., or from rhubarb , which is used in traditional Chinese medicine [Our studies also show that compounds modulating the activity of the lysosomal system are anthraquinones, compounds isolated from aloes of various species  are aloe-emodin  and emodCurrently, in anticancer therapy, attention is paid to combining various types of drugs with anticancer drugs in order to achieve better treatment results. This approach has the potential to reduce drug resistance while providing anti-cancer therapeutic benefits that include the reduction of tumor growth and metastatic potential, the arrest of mitotically active cells, the reduction of cancer stem cell populations, and the induction of apoptosis . Such anLysosomal membrane permeabilization has also been used to restore the sensitivity of lung cancer cells (A549cisR) with acquired chemo-resistance to cisplatin, and is commonly used in advanced lung cancer therapy . AccordiA combination of microtubule-disrupting drugs such as vincristine with lysosome-destabilizing compounds, an example of which is the described siramesine, may be of great importance for cancer therapies. The study showed that combining non-toxic concentrations of vincristine with siramesine effectively induced massive death of cells resistant to the alkaloid (MCF-7 line) .Combination therapy based on a lysosomal system has attracted attention in the context of the use of nintedanib (BIBF1120), which, as a small molecule tyrosine kinase inhibitor (TKI), inhibits the activity of receptor tyrosine kinases (TKs) that include vascular endothelial growth factor receptor (VEGFR 1–3), platelet-derived growth factor receptor (PDGFR-α i-β) and fibroblast growth factor receptor (FGFR 1–3) . BecauseStudies have shown that resistance to nintedanib treatment results from the fact that in lung cancer cells, this compound can be sequestered into lysosomes, where it becomes protonated and trapped. At the same time, it has been shown that the combination of the drug with bafilomycin A1 or chloroquine contributes to the reduction of its accumulation in lysosomes, which should be associated with the inhibition of acidic pH in lysosomes . NintedaIn combination therapy with siramesine, another tyrosine kinase inhibitor, lapatinib, also shows a positive effect in the treatment of prostate cancer. In this case, the intensification of cell death was the result of increased lysosomal membrane permeability, the enhanced production of ROS, and decreased mitochondrial membrane potential. The combined effect of lapatinib and siramesine contributed to the induction of apoptotic death, PARP cleavage and reduced expression of the anti-apoptotic protein Mcl-1 .One of the reasons for the failure of anticancer therapies is the drug resistance of cancer cells, which are characterized by their ability to inhibit programmed cell death by disrupting the apoptosis pathway. Numerous works show that for new therapies in oncological treatment, it is important to activate in cancer cells other apoptosis pathways leading to their death. Such a pathway may be lysosomal cell death. Currently, it is believed that the lysosomal system may be of great importance in chemotherapy because lysosomes in cancer cells are characterized by specific properties that can be used to target anticancer drugs inside them. Their high accumulation inside lysosomes can contribute to the strong sensitization of membranes, resulting in increased permeability, increased leakage of cathepsins into the cytoplasm, induction of apoptotic processes and, consequently, increased cytotoxic effects. From the point of view of modern cancer therapy, it is also extremely important to pay attention to combination therapies in which various types of drugs (including non-cancer drugs), as a consequence of their use with chemotherapeutics, increase cytotoxicity which translates into treatment effectiveness.Understanding the exact mechanisms that induce lysosomal membrane permeability, as well as the mechanisms of action of cathepsins at the cellular level as initiators of cell death, seems to be a highly relevant task for modern oncological treatments.


The skull base has always been regarded as a frontier by surgeons and radiation oncologists since it represents the interface between the intracranial and the extracranial compartment and hosts several critical anatomical structures with an extremely complex and close relationship.Anatomically, the skull base is not a homogeneous region, since it adjoins different regions ; simultaneously, it is crossed by delicate neurovascular structures from the intracranial to the extracranial region.Historically, the sub-specialties of skull base surgery developed from the disciplines involved in the management of the neighboring structures; contemporary anterior skull base surgery mainly derived from rhinology and lateral skull base surgery as an extension of otology, while posterior skull base surgery and the classical open trans-cranial approaches to the anterior and middle fossae were part of the traditional neurosurgical repertoire. As a result, skull base surgery is currently a multidisciplinary field in which neurosurgeons, otorhinolaryngologists, and maxillofacial and plastic-reconstructive surgeons often cooperate; thus, the skull base is no longer considered a barrier for each of these surgical disciplines but a common field of interest .From a pathological perspective, the skull base may be involved by a spectrum of biologically heterogenous neoplasms. In the anterior skull compartment, tumors may arise from the sinonasal epithelium (both mucosa and glands), as well as soft tissues, bone, and cartilage . Among tOver the past decades, skull base surgery has undergone dramatic development towards reduced invasiveness and enhanced surgical precision, which has occurred alongside the evolution of supporting technology. The advent of microsurgery, along with the endoscopic revolution at the beginning of the 21st century, allowed surgeons to gain access to extremely critical anatomical areas and structures, including the clivus, sella and para-sellar regions, cavernous sinus, petrosal temporal bone up to the petroclival junction, cerebellopontine angle, foramen magnum, cranial nerves, and internal carotid artery ,7,8,9.From a surgical perspective, the bony frame of the skull base is neither a barrier nor a constraint for the surgical approach. Rather, it represents a gateway to the critical structures located within the bone of the skull base, since most of the surgical approaches consist in drilling out the bone of the skull base until the target areas are reached. The recent development of virtual surgical planning protocols and image-guided surgery systems have rendered the “surgical corridor” paradigm even more crucial, thus supporting the accurate planning of the surgical trajectory towards the target area and facilitating the anatomical orientation ,12,13,14Along with the current endoscopic techniques and novel extended approaches , which cThe same anatomical and functional principles guiding surgery should also be applied to radiation oncology. In fact, targeting treatment to the involved areas  represents a major challenge for contemporary skull base radiation oncologists. For most resectable craniofacial malignancies, postoperative radiation has a well-established role. Intensity-modulated radiation therapy still represents the most frequently used radiation technique, although the evidence for innovative and potentially less invasive and/or more effective approaches, such as proton therapy and carbon ion irradiation, is promising. In particular, for radioresistant tumors, or in the case of gross residual disease after surgery, the use of proton and carbon ion therapy is now strongly recommended in view of the possibility of escalating the dose and biological effectiveness towards the target while minimizing the irradiation of neurological structures ,18 Figu. This coSystemic therapy represents another promising field of multidisciplinary treatment for skull base tumors, especially in an induction/neoadjuvant setting , with thIn addition to malignant tumors requiring highly demanding multidisciplinary approaches, benign skull base conditions also pose a challenge. Osseous diseases, such as fibrous dysplasia  and osteLastly, inflammatory diseases of the skull base deserve special attention. Osteomyelitis represents a rare but severe condition that is potentially associated with life-threatening complications . The gatJournal of Clinical Medicine entitled “State of the Art-Treatment of skull base diseases” aims to compose a comprehensive picture of the state of the art regarding the multidisciplinary management of skull base diseases in all its complexity, while simultaneously outlining the emerging trends and future development perspectives in this intriguing field of research.This Special Issue of the