Characterization of an in vivo model of neuroinflammation and evaluation of the anti-inflammatory and neuroprotective effects of curcumin as a potential lead compound for the development of new agents useful to treat neuroinflammatory disorders

Neuroinflammation is a complex and multifactorial response of the central nervous system (CNS) to trauma, infection and neurodegenerative diseases orchestrated by specialized immune cells (microglial and astrocytes). In particular, microglia, the main resident immune cells of the CNS, undergo rapid “activation” in response to noxious stimuli, releasing a plethora of inflammatory and potentially neurotoxic soluble factors, such as cytokines [e.g., interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α], chemokines and reactive oxygen and nitrogen species (e.g., nitric oxide). Although an efficient microglial immune response is necessary and critical for proper resolution of pathological events, it is clear that an excessive activation of these cells contributes to neuronal cell damage in neurodegenerative and psychiatric disorders. However, the molecular mechanism(s) by which these cells exert their deleterious effects on neurons remain poorly understood. Recent evidence suggests that microglial cells can be “activated” in response to a systemic inflammatory stimulus. For example, a single peripheral administration of the bacterial endotoxin lipopolysaccharide (LPS), the main component of Gram-negative bacteria walls, in adult mice, can induce microglial activation and an inflammatory state in the CNS that persists long after peripheral stimulus has decline. Identification of molecules which prevent or down-regulate microglial inflammatory responses or direct microglia towards a protective anti-inflammatory phenotype could prove efficacious in neurodegenerative diseases in which inflammation is implicated. Recently, increasing interest has focused on identifying natural compounds with potential inhibitory effects on microglial activation and subsequent inflammatory processes. Among these compounds, curcumin (diferuloylmethane), the main bioactive component isolated from the rhizome of the turmeric plant (Curcuma longa) with multiple pharmacological effects, including anti-inflammatory activities, possesses neuroprotective properties against many neurodegenerative conditions. The principal focus of this doctoral project has been the study of the possible anti-inflammatory effect of curcumin in an in vivo model of neuroinflammation based on a single systemic LPS injection. Young adult mice were intraperitoneally injected with a single dose of LPS (0.5 or 5 mg/kg) or vehicle and then tested for “sickness behavior” (e.g., changes in body weight and food intake), mRNA (real-time RT-PCR) and protein (enzyme-linked immunosorbent assay, ELISA) expression of pro-inflammatory mediators and microglia morphological changes (immune-staining with the microglial marker ionized calcium binding adaptor molecule 1) in different brain areas. Both LPS doses induced a significant decrease in food intake and body weight within the first 4 days, followed by a gradual recovery to control values; however, only 5 mg/kg LPS significantly increased TNF-α, IL-1β, IL-6, COX-2 and iNOS gene expression 2 h post-injection. Pre-treatment with 50 mg/kg curcumin (orally administered by gavage for 2 consecutive days before LPS injection) facilitated the recovery from sickness behavior (anorexia and weight loss), suppressed LPS-induced microglial morphological changes and increased of mRNA levels of TNF-α, IL-1β and COX-2 in all brain areas, while limiting expression of IL-6 and iNOS to more selected brain regions. Possible neuroprotective properties of curcumin were also investigated. In particular, mRNA expression of brain-derived neurotrophic factor (BDNF), known to play a key role in the regulation of neuronal function, as well as in learning and memory processes. Its reduced expression, described in numerous neurodegenerative disorders, contributes to the onset of structural abnormalities and functional damage in the CNS. LPS treatment reduced mRNA levels of BDNF in all brain areas analyzed up to 7 days after treatment. Pre-treatment with 50 mg/kg curcumin restored BDNF gene expression 24 hours after LPS injection, while a higher dose of curcumin (100 mg/kg) completely restored the expression of BDNF 2 hours after the inflammatory stimulus. Finally, behavioral tests demonstrated that LPS induced motor and memory impairments, that persisted even when the inflammatory process had been solved. Moreover, curcumin reversed motor deficits induced by LPS. Taken together, these data show that curcumin can prevent neuroinflammation by modulating the expression of brain pro-inflammatory mediators in vivo and also suggest a potential role of curcumin as a neuroprotective molecule able to restore BDNF levels and behavioral impairments under inflammatory conditions. In conclusion, curcumin represents a promising lead compound to discover new drug candidates, with improved therapeutic efficacy in the treatment of neurodegenerative and age-related diseases with an inflammatory etiology

Human adenocarcinoma cell line sensitivity to essential oil phytocomplexes from pistacia species: A multivariate approach

Principal component analysis (PCA) multivariate analysis was applied to study the cytotoxic activity of essential oils from various species of the Pistacia genus on human tumor cell lines. In particular, the cytotoxic activity of essential oils obtained from P. lentiscus, P. lentiscus var. chia (mastic gum), P. terebinthus, P. vera, and P. integerrima, was screened on three human adenocarcinoma cell lines: MCF-7 (breast), 2008 (ovarian), and LoVo (colon). The results indicate that all the Pistacia phytocomplexes, with the exception of mastic gum oil, induce cytotoxic effects on one or more of the three cell lines. PCA highlighted the presence of different cooperating clusters of bioactive molecules. Cluster variability among species, and even within the same species, could explain some of the differences seen among samples suggesting the presence of both common and species-specific mechanisms. Single molecules from one of the most significant clusters were tested, but only bornyl-acetate presented cytotoxic activity, although at much higher concentrations (IC50 = 138.5 \ub5g/mL) than those present in the essential oils, indicating that understanding of the full biological effect requires a holistic vision of the phytocomplexes with all its constituents

Curcumin Prevents Acute Neuroinflammation and Long-Term Memory Impairment Induced by Systemic Lipopolysaccharide in Mice

Systemic lipopolysaccharide (LPS) induces an acute inflammatory response in the central nervous system (CNS) (\u201cneuroinflammation\u201d) characterized by altered functions of microglial cells, the major resident immune cells of the CNS, and an increased inflammatory profile that can result in long-term neuronal cell damage and severe behavioral and cognitive consequences. Curcumin, a natural compound, exerts CNS anti-inflammatory and neuroprotective functions mainly after chronic treatment. However, its effect after acute treatment has not been well investigated. In the present study, we provide evidence that 50 mg/kg of curcumin, orally administered for 2 consecutive days before a single intraperitoneal injection of a high dose of LPS (5 mg/kg) in young adult mice prevents the CNS immune response. Curcumin, able to enter brain tissue in biologically relevant concentrations, reduced acute and transient microglia activation, pro-inflammatory mediator production, and the behavioral symptoms of sickness. In addition, short-term treatment with curcumin, administered at the time of LPS challenge, anticipated the recovery from memory impairments observed 1 month after the inflammatory stimulus, when mice had completely recovered from the acute neuroinflammation. Together, these results suggest that the preventive effect of curcumin in inhibiting the acute effects of neuroinflammation could be of value in reducing the long-term consequences of brain inflammation, including cognitive deficits such as memory dysfunction

Increased NK Cell Count in Multiple Sclerosis Patients Treated With Dimethyl Fumarate: A 2-Year Longitudinal Study

Background: Dimethyl fumarate (DMF) is a disease-modifying drug for relapsing-remitting multiple sclerosis. Among others, DMF impedes immune activation by shifting the balance between inflammatory and regulatory cell types and by inducing apoptosis-triggered lymphopenia. Although the decrease in lymphocyte count is an early effect of the drug in several patients, the long-term impact on lymphocyte subsets is largely unknown.Methods: We performed a 2-years observational study on total lymphocyte count and subsets thereof by flow cytometry of peripheral blood of 38 multiple sclerosis patients in treatment with DMF. Data were collected at the beginning and after 3, 6, 12, and 24 months of therapy.Results: Total lymphocyte count decreased in relation to time of exposure to DMF. Mean absolute B cell count decreased by 34.1%(p < 0.001) within the first 3months of therapy and then remained stable over time. Mean absolute CD3(+) T cells count decrement reached 47.5% after 12 months of treatment ( p < 0.001). NK cells count showed a heterogeneous trend, increasing by 85.9%( p < 0.001) after 2 years of treatment. CD4(+) T cells and CD8(+) T cells substantially decreased, with a significant increase of CD4(+)/CD8(+) ratio during the first year of therapy.Conclusions: NK cells showed a heterogeneous behavior during DMF treatment with a significant increase over time. Since NK cells may also have a regulatory effect on immune system modulation, their increase during DMF treatment might play a role in the efficacy and safety of the drug

The role of ceus in the evaluation of thyroid cancer : From diagnosis to local staging

Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Ultrasound often represents the first diagnostic step for thyroid nodule evaluation in clinical practice, but baseline US alone is not always effective enough to achieve thyroid nodule characterization. In the last decades new ultrasound techniques, such as CEUS, have been introduced to evaluate thyroid parenchyma as recommended by EFSUMB guidelines, for use in clinical research field, although its role is not yet clear. Several papers show the potential utility of CEUS in the differential diagnosis of benign and malignant thyroid nodules and in the analysis of lymph node involvement in neoplastic pathology. Therefore, we carried out an evaluation of the literature concerning the role of CEUS in three specific areas: the characterization of the thyroid nodule, the evaluation of minimally invasive treatment and loco‐regional staging of the lymph node in proven thyroid cancer. According to evidence reported, CEUS can also play an operative role in nodular thyroid pathology as it is able to guide ablation procedures on thyroid nodule and metastatic lymph nodes, to assess the radicality of surgery, to evaluate disease relapse at the level of the margins of ablated regions and to monitor the clinical evolution of necrotic areas in immediate post‐treatment setting.publishersversionPeer reviewe

Characterization of an in vivo model of neuroinflammation and evaluation of the anti-inflammatory and neuroprotective effects of curcumin as a potential lead compound for the development of new agents useful to treat neuroinflammatory disorders

Neuroinflammation is a complex and multifactorial response of the central nervous system (CNS) to trauma, infection and neurodegenerative diseases orchestrated by specialized immune cells (microglial and astrocytes). In particular, microglia, the main resident immune cells of the CNS, undergo rapid “activation” in response to noxious stimuli, releasing a plethora of inflammatory and potentially neurotoxic soluble factors, such as cytokines [e.g., interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α], chemokines and reactive oxygen and nitrogen species (e.g., nitric oxide). Although an efficient microglial immune response is necessary and critical for proper resolution of pathological events, it is clear that an excessive activation of these cells contributes to neuronal cell damage in neurodegenerative and psychiatric disorders. However, the molecular mechanism(s) by which these cells exert their deleterious effects on neurons remain poorly understood. Recent evidence suggests that microglial cells can be “activated” in response to a systemic inflammatory stimulus. For example, a single peripheral administration of the bacterial endotoxin lipopolysaccharide (LPS), the main component of Gram-negative bacteria walls, in adult mice, can induce microglial activation and an inflammatory state in the CNS that persists long after peripheral stimulus has decline. Identification of molecules which prevent or down-regulate microglial inflammatory responses or direct microglia towards a protective anti-inflammatory phenotype could prove efficacious in neurodegenerative diseases in which inflammation is implicated. Recently, increasing interest has focused on identifying natural compounds with potential inhibitory effects on microglial activation and subsequent inflammatory processes. Among these compounds, curcumin (diferuloylmethane), the main bioactive component isolated from the rhizome of the turmeric plant (Curcuma longa) with multiple pharmacological effects, including anti-inflammatory activities, possesses neuroprotective properties against many neurodegenerative conditions. The principal focus of this doctoral project has been the study of the possible anti-inflammatory effect of curcumin in an in vivo model of neuroinflammation based on a single systemic LPS injection. Young adult mice were intraperitoneally injected with a single dose of LPS (0.5 or 5 mg/kg) or vehicle and then tested for “sickness behavior” (e.g., changes in body weight and food intake), mRNA (real-time RT-PCR) and protein (enzyme-linked immunosorbent assay, ELISA) expression of pro-inflammatory mediators and microglia morphological changes (immune-staining with the microglial marker ionized calcium binding adaptor molecule 1) in different brain areas. Both LPS doses induced a significant decrease in food intake and body weight within the first 4 days, followed by a gradual recovery to control values; however, only 5 mg/kg LPS significantly increased TNF-α, IL-1β, IL-6, COX-2 and iNOS gene expression 2 h post-injection. Pre-treatment with 50 mg/kg curcumin (orally administered by gavage for 2 consecutive days before LPS injection) facilitated the recovery from sickness behavior (anorexia and weight loss), suppressed LPS-induced microglial morphological changes and increased of mRNA levels of TNF-α, IL-1β and COX-2 in all brain areas, while limiting expression of IL-6 and iNOS to more selected brain regions. Possible neuroprotective properties of curcumin were also investigated. In particular, mRNA expression of brain-derived neurotrophic factor (BDNF), known to play a key role in the regulation of neuronal function, as well as in learning and memory processes. Its reduced expression, described in numerous neurodegenerative disorders, contributes to the onset of structural abnormalities and functional damage in the CNS. LPS treatment reduced mRNA levels of BDNF in all brain areas analyzed up to 7 days after treatment. Pre-treatment with 50 mg/kg curcumin restored BDNF gene expression 24 hours after LPS injection, while a higher dose of curcumin (100 mg/kg) completely restored the expression of BDNF 2 hours after the inflammatory stimulus. Finally, behavioral tests demonstrated that LPS induced motor and memory impairments, that persisted even when the inflammatory process had been solved. Moreover, curcumin reversed motor deficits induced by LPS. Taken together, these data show that curcumin can prevent neuroinflammation by modulating the expression of brain pro-inflammatory mediators in vivo and also suggest a potential role of curcumin as a neuroprotective molecule able to restore BDNF levels and behavioral impairments under inflammatory conditions. In conclusion, curcumin represents a promising lead compound to discover new drug candidates, with improved therapeutic efficacy in the treatment of neurodegenerative and age-related diseases with an inflammatory etiology.La neuroinfiammazione è una risposta complessa e multifattoriale a diversi tipi di stimoli nocivi come le infezioni virali, batteriche o traumi. é noto che in questo processo le cellule microgliali giochino in ruolo chiavenel bilancio tra la risoluzione del processo infiammatorio o la progressione di questo ferso un profilo neurodegenerativo. Nel nostro lavoro abbiamo valutato gli effetti antiinfiammatori e neuroprotettivi di curcumina in un modello in vivo di neuroinfiammazione creato mediante una singola iniezione di lipopolisaccaride (LPS) intraperitoneale ad una dose di 5 mg/Kg. L'effetto di curcumina è stato valutato sul profilo citochinico, su molecole proinfimmatorie come COX-2 e iNOS su molecole neuroprotettive come il BDNF mediante saggi di RT-PCR, immunoistochimica per evidenziare l'effetto di curcumina sull'attivazione microgliale e sulla sopravvivenza dei neuroni dopaminergici in substantia nigra, attraverso l'uso di test comportamentali ai fini di valutare l'effetto protettivo di curcumina sull'attività locomotoria e la memoria dichiarativa. In cocnlusione, la curcumina è risultata essere un valido composto con proprietà antiinfiammatorie e neuroprotettive e rappresenta una molecola chiave per lo sviuppo di nuovi composti con aumentata efficacia terapeutica nel trattamento e la prevenzione di patologie neurodegenerative su base infiammatoria

Il sistema tioredossinico come target di nuovi complessi metallici nella terapia anticancro

Thioredoxin system acts as anticancer target for metal-based drug

A model of systemic inflammation to study neuroinflammation

Increasing evidence suggests that neurodegeneration occurs in part because the environment is affected during disease in a cascade of processes collectively termed neuroinflammation. This is a reactive response of the central nervous system against noxious elements that interfere with tissue homeostasis. Neuroinflammation is mediated by inflammatory molecules released by microglial cells. Understanding and controlling interactions between the immune system and microglial activation might represent the key to prevent or delay the onset of central nervous system diseases. This chapter details techniques to generate and characterize an in vivo model of neuroinflammation based on a single intraperitoneal injection of lipopolysaccharide, which can be used to understand the wide variety of cellular and molecular mechanisms of neuroinflammation, as well as to identify new therapies by testing the anti-inflammatory properties of synthetic and natural molecules

Un modello fisico-matematico del funzionamento della macchina di Stirling.

[no abstract available

Vitamin D Physiology, Deficiency, Genetic Influence, and the Effects of Daily vs. Bolus Doses of Vitamin D on Overall Health: A Clinical Approach

Vitamin D is a pleiotropic hormone that plays a vital role in regulating bone growth, maintaining calcium and phosphate homeostasis, modulating immune function, and a wide range of other pleiotrophic actions in humans, which have increased the attention for its clinical applications. Despite its importance, vitamin D deficiency is prevalent worldwide and is related to a range of pathophysiological conditions, including an increased risk of osteoporosis and chronic and autoimmune diseases. The recommended daily doses of vitamin D vary depending on genetics, age, sex, and health status, with specific doses recommended for infants, children, adults, and those at increased risk of deficiency or specific health conditions. Maintaining adequate vitamin D levels is essential for optimal health, and together with sun exposure, appropriate supplementation strategies can help achieve this goal. Vitamin D supplementation is commonly used to maintain adequate levels, and the optimal administration strategy, such as a daily dose vs. a bolus, is still being investigated. This review aims to understand vitamin D physiology and the impact of relevant vitamin D polymorphisms and to evaluate the role of a daily dose versus a bolus in maintaining optimal vitamin D levels and clinical health outcomes. It also provides suggested clinical guidelines for clinicians based on the most recent scientific evidence