The neural substrate of spatial memory stabilization depends on the distribution of the training sessions.

SignificanceDistributed training has long been known to lead to more robust memory formation as compared to massed training. Using the water maze, a well-established task for assessing memory in laboratory rodents, we found that distributed and massed training differentially engage the dorsolateral and dorsomedial striatum, and optogenetic priming of dorsolateral striatum can artificially increase the robustness of massed training to the level of distributed training. Overall, our findings demonstrate that spatial memory consolidation engages different neural substrates depending on the training regimen, identifying a therapeutic avenue for memory enhancement.

BV2-derived extracellular vesicles modulate microglia inflammatory profile, neuronal plasticity, and behavioural performances in late adult mice.

BACKGROUND: During aging, both the brain and the immune system undergo a progressive impairment of physiological functions. Microglia, the immunocompetent cells of the central nervous system, shift towards a chronic mild inflammatory state that impacts brain homeostasis. Extracellular vesicles (EVs) released by microglia transport packages of molecular information that mirror the inflammatory status of donor cells and modulate the inflammatory phenotype of recipient microglia and other cell types. RESULTS: We demonstrated that intranasal administration of EVs derived from microglial-like BV2 cells to late adult mice (16-20 months of age) shifts microglia toward a "juvenile" morphology affecting their inflammatory profile. Mice treated with BV2-derived EVs have a reduction of anxiety-like behavior and an increased spatial learning, with sex-dependent differences. Further, BV2-derived EVs increased neuronal plasticity both in male and female mice. These findings suggest the involvement of microglial cells in vesicles-mediated anti-aging effect. CONCLUSIONS: Our data indicate that BV2-derived EVs could represent a resource to slow down age-dependent inflammation in the mouse brain.

Adult hippocampal neurogenesis and social behavioural deficits in the R451C Neuroligin3 mouse model of autism are reverted by the antidepressant fluoxetine.

Neuron generation persists throughout life in the hippocampus but is altered in animal models of neurological and neuropsychiatric diseases, suggesting that disease-associated decline in cognitive and emotional hippocampal-dependent behaviours might be functionally linked with dysregulation of postnatal neurogenesis. Depletion of the adult neural stem/progenitor cell (NSPCs) pool and neurogenic decline have been recently described in mice expressing synaptic susceptibility genes associated with autism spectrum disorder (ASDs). To gain further insight into mechanisms regulating neurogenesis in mice carrying mutations in synaptic genes related to monogenic ASDs, we used the R451C Neuroligin3 knock-in (Nlgn3 KI) mouse, which is characterized by structural brain abnormalities, deficits in synaptic functions and reduced sociability. We show that the number of adult-born neurons, but not the size of the NSPC pool, was reduced in the ventral dentate gyrus in knock-in mice. Notably, this neurogenic decline was rescued by daily injecting mice with 10 mg/Kg of the antidepressant fluoxetine for 20 consecutive days. Sustained treatment also improved KI mice's sociability and increased the number of c-Fos active adult-born neurons, compared with vehicle-injected KI mice. Our study uncovers neurogenesis-mediated alterations in the brain of R451C KI mouse, showing that the R451C Nlgn3 mutation leads to lasting, albeit pharmacologically reversible, changes in the brain, affecting neuron formation in the adult hippocampus. Our results suggest that fluoxetine can ameliorate social behaviour in KI mice, at least in part, by rescuing adult hippocampal neurogenesis, which may be relevant for the pharmacological treatment of ASDs.

Stress-induced strain and brain region-specific activation of LINE-1 transposons in adult mice.

Transposable elements (TEs) are conserved mobile genetic elements that are highly abundant in most eukaryotic genomes. Although the exact function of TEs is still largely unknown, it is increasingly clear that they are significantly modulated in response to stress in a wide range of organisms, either directly or indirectly through regulation of epigenetic silencing. We investigated the effect of repeated restraint stress (2 h a day, for 5 d) on transcription levels of LINE-1 (L1) retrotransposon in the brain of inbred BALB/c, DBA/2, C57BL/6N, and outbred CD1 mice. Repeated restraint stress induced strain and brain region-specific modulation of L1 activity. We observed a significant derepression of L1 transcription in the hippocampus (HIPP) of BALB/c mice and a significant downregulation in the hippocampus of C57BL/6N mice. No significant change in L1 expression was found in the other strains and brain regions. These findings indicate in mice the control of transposons expression as an additional mechanism in stress-induced pathophysiological responses, demonstrating that their regulation is highly dependent on the strain genetic background and the brain region. Lay summary Hippocampal expression of the transposon L1 is significantly altered by repeated restraint stress in mice. L1 modulation is not only region specific, but also strain dependent, suggesting that the genetic background is an important determinant of L1 response to environmental stimuli.

Active integration of glutamatergic input to the inferior olive generates bidirectional postsynaptic potentials.

KEY POINTS: We establish experimental preparations for optogenetic investigation of glutamatergic input to the inferior olive. Neurones in the principal olivary nucleus receive monosynaptic extra-somatic glutamatergic input from the neocortex. Glutamatergic inputs to neurones in the inferior olive generate bidirectional postsynaptic potentials (PSPs), with a fast excitatory component followed by a slower inhibitory component. Small conductance calcium-activated potassium (SK) channels are required for the slow inhibitory component of glutamatergic PSPs and oppose temporal summation of inputs at intervals ≤ 20 ms. Active integration of synaptic input within the inferior olive may play a central role in control of olivo-cerebellar climbing fibre signals. ABSTRACT: The inferior olive plays a critical role in motor coordination and learning by integrating diverse afferent signals to generate climbing fibre inputs to the cerebellar cortex. While it is well established that climbing fibre signals are important for motor coordination, the mechanisms by which neurones in the inferior olive integrate synaptic inputs and the roles of particular ion channels are unclear. Here, we test the hypothesis that neurones in the inferior olive actively integrate glutamatergic synaptic inputs. We demonstrate that optogenetically activated long-range synaptic inputs to the inferior olive, including projections from the motor cortex, generate rapid excitatory potentials followed by slower inhibitory potentials. Synaptic projections from the motor cortex preferentially target the principal olivary nucleus. We show that inhibitory and excitatory components of the bidirectional synaptic potentials are dependent upon AMPA (GluA) receptors, are GABAA independent, and originate from the same presynaptic axons. Consistent with models that predict active integration of synaptic inputs by inferior olive neurones, we find that the inhibitory component is reduced by blocking large conductance calcium-activated potassium channels with iberiotoxin, and is abolished by blocking small conductance calcium-activated potassium channels with apamin. Summation of excitatory components of synaptic responses to inputs at intervals ≤ 20 ms is increased by apamin, suggesting a role for the inhibitory component of glutamatergic responses in temporal integration. Our results indicate that neurones in the inferior olive implement novel rules for synaptic integration and suggest new principles for the contribution of inferior olive neurones to coordinated motor behaviours.

[The burden of informal caregivers of multiple sclerosis patients: a pilot study]. / Il Burden del caregiver informale dei malati di Sclerosi Multipla: Studio Pilota.

AIM: The main objective of the study was to measure caregiver burden among informal caregivers assisting people with multiple sclerosis (MS). BACKGROUND: Thirty percent of people with multiple sclerosis require some form of home assistance and 80% of such care is provided by informal caregivers. The quality of life of informal caregivers of patients with multiple sclerosis decreases with increasing physical and depressive symptoms of their loved ones. This can have effects on stress levels, physical and mental health, quality of life, and mortality of informal caregivers. DESIGN: A descriptive correlational study was performed in a sample of informal caregivers of MS patients. METHODS: The study enrolled 41 informal caregivers of MS patients at a regional reference center in Rome, Italy. Data collection took place in the period between May and September 2016. A paper questionnaire was used, comprising a socio-demographic section and a "Caregiver Burden Inventory" section. RESULTS: The subjective burden perceived by informal caregivers of patients with MS is moderate. Socio-demographic characteristics were not found to influence burden of caregivers. CONCLUSIONS: It is important for nursing professionals to consider and evaluate the burden of informal caregivers of MS patients as they are valuable allies in the management of these patients.

Efficacy of Radio Electric Asymmetric Conveyer Neuro Psycho Physical Optimization - Brain Wave Optimization-Gamma (REAC NPPO BWO-G) Treatment in Neurovegetative Dysfunction: A Case Report of Enhanced Cognitive Processing and Stress Resilience.

Neurovegetative dysfunction from chronic stress impairs cognition, emotional regulation, and quality of life, with limited relief from conventional therapies. The Radio Electric Asymmetric Conveyer (REAC) Neuro Psycho Physical Optimization - Brain Wave Optimization-Gamma (NPPO BWO-G) offers a novel non-invasive approach to restore autonomic balance through brain modulation. This case involves a 63-year-old businessman with atrial fibrillation, fatigue, cognitive decline, and sleep issues. Pre-treatment quantitative electroencephalogram (qEEG) showed low brain activity and excess delta rhythms. After REAC NPPO BWO-G sessions, the patient experienced improved brainwave patterns, cognitive clarity, stress management, and reduced fatigue. These results highlight its potential as a promising treatment for stress-related neurovegetative dysfunction, warranting further study.

Distinct roles of medial prefrontal cortex subregions in the consolidation and recall of remote spatial memories.

It is a common belief that memories, over time, become progressively independent of the hippocampus and are gradually stored in cortical areas. This view is mainly based on evidence showing that prefrontal cortex (PFC) manipulations impair the retrieval of remote memories, while hippocampal inhibition does not. More controversial is whether activity in the medial prefrontal cortex (mPFC) is required immediately after learning to initiate consolidation. Another question concerns functional differences among PFC subregions in forming and storing remote memories.To address these issues, we directly contrasted the effects of loss-of-function manipulations of the anterior cingulate cortex (aCC) and the ventromedial PFC, which includes the infralimbic (IL) and prelimbic (PL) cortices, before testing and immediately after training on the ability of CD1 mice to recall the hidden platform location in the Morris water maze. We injected an AAV carrying the hM4Di receptor into the PL-IL or aCC. Interestingly, pre-test administrations of clozapine-N-oxide (CNO) (3 mg/kg) revealed that the aCC, but not the PL-IL, was necessary to recall remote spatial information. Furthermore, systemic post-training administration of CNO impaired memory recall at remote, but not recent, time points in both groups. These findings revealed a functional dissociation between the two prefrontal areas, demonstrating that both the PL-IL and the aCC are involved in early consolidation of remote spatial memories, but only the aCC is engaged in their recall.Significant statement Contrasting the two main components of the prefrontal cortex, the anterior cingulate (aCC) and the ventro medial component, which includes the prelimbic and the infralimbic areas (PL-IL), we investigated their respective contributions to the recall of remote spatial memory and the consolidation of both recent and remote memories. The results demonstrated an interesting dissociation between these regions. Loss-of-function manipulation of the aCC, but not the PL-IL, impaired the recall of remote spatial memory. However, both subdivisions contributed to the consolidation of remote spatial memory. These findings provide new insight into the circuit dynamic involved in the formation of stable remote memories, suggesting a regulatory role of the PL-IL in selecting relevant information from storage.

Neurobiological modulation with REAC technology: enhancing pain, depression, anxiety, stress, and quality of life in post-polio syndrome subjects.

Post-polio syndrome (PPS) brings new challenges for polio survivors, including muscle decline, pain, depression, and diminished quality of life. This study explored the potential of REAC neuromodulatory treatments to ease pain, improve mood, and enhance quality of life in PPS patients. 17 individuals with PPS (average age 54.8) received three REAC treatments: Neuro Postural Optimization, Neuro Psycho Physical Optimization, and Neuro Psycho Physical Optimization-Cervico Brachial. Pain, depression, anxiety, stress, and quality of life were assessed before and after using established scales. REAC treatments significantly reduced pain across various dimensions, along with depression, anxiety, and stress levels. Additionally, patients reported improved physical and psychological quality of life. This study suggests REAC neuromodulatory treatments as a promising non-invasive option to improve pain, emotional well-being, and quality of life in individuals with PPS.

Polymer-lipid hybrid nanomedicines to deliver siRNA in and against glioblastoma cells.

Small interfering RNA (siRNA) holds great potential to treat many difficult-to-treat diseases, but its delivery remains the central challenge. This study aimed at investigating the suitability of polymer-lipid hybrid nanomedicines (HNMeds) as novel siRNA delivery platforms for locoregional therapy of glioblastoma. Two HNMed formulations were developed from poly(lactic-co-glycolic acid) polymer and a cationic lipid: 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 3ß-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol (DC-Chol). After characterization of the HNMeds, a model siRNA was complexed onto their surface to form HNMed/siRNA complexes. The physicochemical properties and siRNA binding ability of complexes were assessed over a range of nitrogen-to-phosphate (N/P) ratios to optimize the formulations. At the optimal N/P ratio of 10, complexes effectively bound siRNA and improved its protection from enzymatic degradation. Using the NIH3T3 mouse fibroblast cell line, DOTAP-based HNMeds were shown to possess higher cytocompatibility in vitro over the DC-Chol-based ones. As proof-of-concept, uptake and bioefficacy of formulations were also assessed in vitro on U87MG human glioblastoma cell line expressing luciferase gene. Complexes were able to deliver anti-luciferase siRNA and induce a remarkable suppression of gene expression. Noteworthy, the effect of DOTAP-based formulation was not only about three-times higher than DC-Chol-based one, but also comparable to lipofectamine model transfection reagent. These findings set the basis to exploit this nanosystem for silencing relevant GB-related genes in further in vitro and in vivo studies.

HCN1 channels in cerebellar Purkinje cells promote late stages of learning and constrain synaptic inhibition.

Neural computations rely on ion channels that modify neuronal responses to synaptic inputs. While single cell recordings suggest diverse and neurone type-specific computational functions for HCN1 channels, their behavioural roles in any single neurone type are not clear. Using a battery of behavioural assays, including analysis of motor learning in vestibulo-ocular reflex and rotarod tests, we find that deletion of HCN1 channels from cerebellar Purkinje cells selectively impairs late stages of motor learning. Because deletion of HCN1 modifies only a subset of behaviours involving Purkinje cells, we asked whether the channel also has functional specificity at a cellular level. We find that HCN1 channels in cerebellar Purkinje cells reduce the duration of inhibitory synaptic responses but, in the absence of membrane hyperpolarization, do not affect responses to excitatory inputs. Our results indicate that manipulation of subthreshold computation in a single neurone type causes specific modifications to behaviour.

Ventral striatal plasticity and spatial memory.

Spatial memory formation is a dynamic process requiring a series of cellular and molecular steps, such as gene expression and protein translation, leading to morphological changes that have been envisaged as the structural bases for the engram. Despite the role suggested for medial temporal lobe plasticity in spatial memory, recent behavioral observations implicate specific components of the striatal complex in spatial information processing. However, the potential occurrence of neural plasticity within this structure after spatial learning has never been investigated. In this study we demonstrate that blockade of cAMP response element binding protein-induced transcription or inhibition of protein synthesis or extracellular proteolytic activity in the ventral striatum impairs long-term spatial memory. These findings demonstrate that, in the ventral striatum, similarly to what happens in the hippocampus, several key molecular events crucial for the expression of neural plasticity are required in the early stages of spatial memory formation.

REAC Noninvasive Neurobiological Stimulation in Autism Spectrum Disorder for Alleviating Stress Impact.

Objectives: Autism spectrum disorder (ASD) symptoms can become more evident because of different factors. Among these, depression, anxiety, and stress play an important role. Additionally, several studies have revealed the impact of the COVID-19 pandemic on participants with ASD. In previous studies, two noninvasive neurobiological stimulation treatments with radio electric asymmetric conveyer (REAC) technology, called neuropostural optimization (NPO) and neuropsychophysical optimization (NPPO), were shown to be effective in improving the subjective response to environmental stressors in the general population and in ASD population. Based on the proven efficacy of REAC NPO and NPPOs treatments in alleviating anxiety, stress, and depression, the purpose of this study is to verify how these treatments can reduce the severity of ASD symptoms expression, which is aggravated by depression, anxiety, and stress. The treatments' effects were perceived by caregivers and assessed by the Autism Treatment Evaluation Checklist (ATEC). Methods: This study involved 46 children with a previous diagnosis of ASD made using the Autism Diagnostic Observation Schedule and Autism Diagnostic Interview-Revised. The participants received one session of NPO treatment and one NPPOs treatment cycle of 18 sessions, administered within approximately 3 weeks. The Autism Treatment Evaluation Checklist (ATEC) was used to evaluate the efficacy of the REAC treatments. ATEC allows to evaluate four clusters (speech or language communication; sociability; sensory or cognitive awareness; and health/physical/behavior) through a numerical scale that measures increasing levels of ASD severity. Results: The comparison between the scores of the ATEC administered pre- and post-REAC treatments highlighted an improvement of ASD symptoms in each of the four clusters of ATEC. Conclusions: The results confirm the usefulness of REAC treatments to optimize the individual response to environmental stressors and reduce the symptomatic expression and deficits present in ASD.

Specific patterns of neural activity in the hippocampus after massed or distributed spatial training.

Training with long inter-session intervals, termed distributed training, has long been known to be superior to training with short intervals, termed massed training. In the present study we compared c-Fos expression after massed and distributed training protocols in the Morris water maze to outline possible differences in the learning-induced pattern of neural activation in the dorsal CA1 in the two training conditions. The results demonstrate that training and time lags between learning opportunities had an impact on the pattern of neuronal activity in the dorsal CA1. Mice trained with the distributed protocol showed sustained neuronal activity in the postero-distal component of the dorsal CA1. In parallel, in trained mice we found more active cells that tended to constitute spatially restricted clusters, whose degree increased with the increase in the time lags between learning trials. Moreover, activated cell assemblies demonstrated increased stability in their spatial organization after distributed as compared to massed training or control condition. Finally, using a machine learning algorithm we found that differences in the number of c-Fos positive cells and their location in the dorsal CA1 could be predictive of the training protocol used. These results suggest that the topographic organization and the spatial location of learning activated cell assemblies might be critical to promote the increased stability of the memory trace induced by distributed training.

Improving Functional Abilities in Children and Adolescents with Autism Spectrum Disorder Using Non-Invasive REAC Neuro Psycho Physical Optimization Treatments: A PEDI-CAT Study.

Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder that affects communication, social interaction, and behavior. Non-invasive neuromodulation techniques, such as radioelectric asymmetric conveyer (REAC) technology, have gained attention for their potential to improve the endogenous bioelectric activity (EBA) and neurobiological processes underlying ASD. Neuro Postural Optimization (NPO) and Neuro Psycho Physical Optimization (NPPO) treatments are non-invasive and painless neuromodulation treatments that utilize REAC technology and have shown promising results in improving the symptoms of ASD. This study aimed to evaluate the effects of NPO and NPPO treatments on functional abilities in children and adolescents with ASD using the Pediatric Evaluation of Disability Inventory-Computer Adaptive Test (PEDI-CAT). The study consisted of 27 children and adolescents with ASD who underwent a single session of NPO followed by 18 sessions of NPPO treatment over a period of one week. The results showed significant improvements in the children's and adolescents' functional abilities across all domains of the PEDI-CAT. These findings suggest that NPO and NPPO may be effective treatments for improving functional abilities in children and adolescents with ASD.

REAC Neurobiological Modulation as a Precision Medicine Treatment for Fibromyalgia.

Fibromyalgia syndrome (FS) is a disorder characterized by widespread musculoskeletal pain and psychopathological symptoms, often associated with central pain modulation failure and dysfunctional adaptive responses to environmental stress. The Radio Electric Asymmetric Conveyer (REAC) technology is a neuromodulation technology. The aim of this study was to evaluate the effects of some REAC treatments on psychomotor responses and quality of life in 37 patients with FS. Tests were conducted before and after a single session of Neuro Postural Optimization and after a cycle of 18 sessions of Neuro Psycho Physical Optimization (NPPO), using evaluation of the functional dysmetria (FD) phenomenon, Sitting and Standing (SS), Time Up and Go (TUG) tests for motor evaluation, Fibromyalgia Impact Questionnaire (FIQ) for quality of life. The data were statistically analyzed, and the results showed a statistically significant improvement in motor response and quality of life parameters, including pain, as well as reduced FD measures in all participants. The study concludes that the neurobiological balance established by the REAC therapeutic protocols NPO and NPPO improved the dysfunctional adaptive state caused by environmental and exposomal stress in FS patients, leading to an improvement in psychomotor responses and quality of life. The findings suggest that REAC treatments could be an effective approach for FS patients, reducing the excessive use of analgesic drugs and improving daily activities.

Spatial memory, plasticity and nucleus accumbens.

Research on the function of the nucleus accumbens, the most ventral component of the striatal complex, has traditionally focused on locomotor activity, reward, motivation and addiction. However, based on the existence of projections to the nucleus accumbens from the allocortical regions involved in spatial navigation, it has been suggested that this structure plays a role in spatial learning and memory. Lesion and neuropharmacological studies confirm this view, also revealing the complex dynamics of the receptors involved in these processes. Moreover, the effects of post-training intra-nucleus accumbens drug administrations demonstrate the necessity of off-line neural activity within this structure in order to consolidate spatial memory. Blockade of molecular processes implicated in synaptic plasticity, such as cAMP response element-binding protein (CREB)-induced transcription or extracellular matrix remodeling, provides further experimental support to this hypothesis. These observations imply that experience-dependent synaptic plasticity responsible for long-term stabilization of spatial information might occur within the nucleus accumbens, similarly to what has been observed in the hippocampus. This suggests that a comprehensive understanding of spatial memory processing should be viewed in the context of a wider neural circuit.

Radio Electric Asymmetric Conveyer (REAC) Neurobiological Stimulation Treatments in Dysfunctional Motor Behavior in Flail Arm Syndrome: A Case Report.

Flail arm syndrome (FAS) is a variant of amyotrophic lateral sclerosis (ALS) that manifests itself with the progressive loss of motor control of the upper limbs starting from the proximal part. Both electrophysiological and magnetic resonance studies have shown that functional alterations in the subcortical structures, cerebellum, and cortex are present in this pathology. These alterations appear to play a significant component in determining cognitive, motor, and behavioral effects. To try to modulate these alterations, in this case report, we used three noninvasive and specific neuromodulation treatments of the Radio Electric Asymmetric Conveyer (REAC) technology. The Neuro Postural Optimization (NPO), the Neuro Psycho Physical Optimization (NPPO), and the Neuro Psycho Physical Optimization Cervico-Brachial (NPPO-CB) with the aim of improving motor control, depression, anxiety, and stress. At the end of the treatment cycle that lasted five consecutive days, the patient regained the ability to raise his arms, a capacity he had lost for several months. This case demonstrates that REAC neurobiological modulation treatments aimed at improving dysfunctional neuropsychomotor behavior (DNPMB) can be useful in highlighting and reducing these components, allowing for better evaluation of the real neurodegenerative damage and determination of a better quality of life for these patients.

Calcific Tendinitis of the Shoulder: A Neuro-Psychomotor Behavioral Diagnostic and Therapeutic Approach With Radioelectric Asymmetric Conveyer Neurobiological Stimulation Treatments.

Calcific tendinitis of the shoulder (CTS) is one of the pathological conditions that most often affects the shoulder and consists of a calcium deposit that settles within the tendon tissue of the rotator cuff. The scientific literature has long highlighted the impact of anxiety, stress, and depression on CTS. The goal of this case report is to highlight how the emotional state of patients and their neuro-psychomotor behavior induce a state of constant muscular tension which, through the physical phenomenon of piezoelectricity, causes calcium salts to precipitate and form calcifications. Therefore, stress, anxiety, and depression are likely factors underlying the etiopathogenesis of CTS. Consistent with this interpretation, this report presents five cases of CTS treated with three specific neurobiological stimulation treatments using the radioelectric asymmetric conveyer (REAC) technology, which has demonstrated its effectiveness on alterations in postural attitude intended as neuro-psychomotor behavior, anxiety, stress, and depression, as well as on autonomic and metabolic alterations of the tissues at a local level. The results presented suggest that this approach may be useful in the treatment and prevention of CTS.

Applications of the ROS-Responsive Thioketal Linker for the Production of Smart Nanomedicines.

Reactive oxygen species (ROS)-sensitive drug delivery systems (DDS) specifically responding to altered levels of ROS in the pathological microenvironment have emerged as an effective means to enhance the pharmaceutical efficacy of conventional nanomedicines, while simultaneously reducing side effects. In particular, the use of the biocompatible, biodegradable, and non-toxic ROS-responsive thioketal (TK) functional group in the design of smart DDS has grown exponentially in recent years. In the design of TK-based DDS, different technological uses of TK have been proposed to overcome the major limitations of conventional DDS counterparts including uncontrolled drug release and off-target effects. This review will focus on the different technological uses of TK-based biomaterials in smart nanomedicines by using it as a linker to connect a drug on the surface of nanoparticles, form prodrugs, as a core component of the DDS to directly control its structure, to control the opening of drug-releasing gates or to change the conformation of the nano-systems. A comprehensive view of the various uses of TK may allow researchers to exploit this reactive linker more consciously while designing nanomedicines to be more effective with improved disease-targeting ability, providing novel therapeutic opportunities in the treatment of many diseases.