A Facile Access to Green Fluorescent Albumin Derivatives.

A Morita-Baylis-Hillman Adduct (MBHA) derivative bearing a triphenylamine moiety was found to react with human serum albumin (HSA) shifting its emission from the blue to the green-yellow thus leading to green fluorescent albumin (GFA) derivatives and enlarging the platform of probes for aggregation-induced fluorescent-based detection techniques. A possible interaction of MBHA derivative 7 with a lipophilic pocket within the HSA structure was suggested by docking studies. DLS experiments showed that the reaction with HSA induce a conformational change of the protein contributing to the aggregation process of GFA derivatives. The results of investigations on the biological properties suggested that GFA retained the ability of binding drug molecules such as warfarin and diazepam. Finally, cytotoxicity evaluation studies suggested that, although the MBHA derivative 7 at 0.1 µg/mL affected the percentage of cell viability in comparison to the negative control, it cannot be considered cytotoxic, whereas at all the other concentrations≥0.5 µg/mL resulted cytotoxic at different extent.

Investigation on Novel E/Z 2-Benzylideneindan-1-One-Based Photoswitches with AChE and MAO-B Dual Inhibitory Activity.

The multitarget therapeutic strategy, as opposed to the more traditional 'one disease-one target-one drug', may hold promise in treating multifactorial neurodegenerative syndromes, such as Alzheimer's disease (AD) and related dementias. Recently, combining a photopharmacology approach with the multitarget-directed ligand (MTDL) design strategy, we disclosed a novel donepezil-like compound, namely 2-(4-((diethylamino)methyl)benzylidene)-5-methoxy-2,3-dihydro-1H-inden-1-one (1a), which in the E isomeric form (and about tenfold less in the UV-B photo-induced isomer Z) showed the best activity as dual inhibitor of the AD-related targets acetylcholinesterase (AChE) and monoamine oxidase B (MAO-B). Herein, we investigated further photoisomerizable 2-benzylideneindan-1-one analogs 1b-h with the unconjugated tertiary amino moiety bearing alkyls of different bulkiness and lipophilicity. For each compound, the thermal stable E geometric isomer, along with the E/Z mixture as produced by UV-B light irradiation in the photostationary state (PSS, 75% Z), was investigated for the inhibition of human ChEs and MAOs. The pure E-isomer of the N-benzyl(ethyl)amino analog 1h achieved low nanomolar AChE and high nanomolar MAO-B inhibition potencies (IC50s 39 and 355 nM, respectively), whereas photoisomerization to the Z isomer (75% Z in the PSS mixture) resulted in a decrease (about 30%) of AChE inhibitory potency, and not in the MAO-B one. Molecular docking studies were performed to rationalize the different E/Z selectivity of 1h toward the two target enzymes.

Xanthopsin-Like Systems via Site-Specific Click-Functionalization of a Retinoic Acid Binding Protein.

The use of light-responsive proteins to control both living or synthetic cells, is at the core of the expanding fields of optogenetics and synthetic biology. It is thus apparent that a richer reaction toolbox for the preparation of such systems is of fundamental importance. Here, we provide a proof-of-principle demonstration that Morita-Baylis-Hillman adducts can be employed to perform a facile site-specific, irreversible and diastereoselective click-functionalization of a lysine residue buried into a lipophilic binding pocket and yielding an unnatural chromophore with an extended π-system. In doing so we effectively open the path to the in vitro preparation of a library of synthetic proteins structurally reminiscent of xanthopsin eubacterial photoreceptors. We argue that such a library, made of variable unnatural chromophores inserted in an easy-to-mutate and crystallize retinoic acid transporter, significantly expand the scope of the recently introduced rhodopsin mimics as both optogenetic and "lab-on-a-molecule" tools.

Design, Synthesis and Characterization of a Visible-Light-Sensitive Molecular Switch and Its PEGylation Towards a Self-Assembling Molecule.

HBDI-like chromophores represent a novel set of biomimetic switches mimicking the fluorophore of the green fluorescent protein that are currently studied with the hope to expand the molecular switch/motor toolbox. However, until now members capable of absorbing visible light in their neutral (i. e. non-anionic) form have not been reported. In this contribution we report the preparation of an HBDI-like chromophore based on a 3-phenylbenzofulvene scaffold capable of absorbing blue light and photoisomerizing on the picosecond timescale. More specifically, we show that double-bond photoisomerization occurs in both the E-to-Z and Z-to-E directions and that these can be controlled by irradiating with blue and UV light, respectively. Finally, as a preliminary applicative result, we report the incorporation of the chromophore in an amphiphilic molecule and demonstrate the formation of a visible-light-sensitive nanoaggregated state in water.

Investigating the relationship between orthorexia nervosa and autistic traits in a university population.

BACKGROUND: Orthorexia nervosa (ON) is an emerging condition featuring restrictive eating behaviors on the basis of subjective beliefs about food healthiness. Many authors have stressed the similarities between ON and anorexia nervosa (AN) in both cognitive and behavioral patterns. Despite that, while the link between AN and female autism presentations is well known in the literature, no study has yet investigated the relationship between ON and autism spectrum. This work aims to investigate the relationship between ON and autistic traits in a university population. METHODS: An e-mail invitation was sent to all the students and University workers of University of Pisa. Subjects were asked to fulfill the ORTO-15 and the Adult Autism Subthreshold spectrum (AdAS spectrum) questionnaires. RESULTS: A total of 2426 subjects joined the survey: 623 subjects (26.3%) reported a score associated with significant orthorexic symptoms according to ORTO-15 (ON group), while 1789 subjects (73.7%) did not report ON symptomatology and were considered as healthy controls (HC). The ON group scored significantly higher on almost all AdAS spectrum domains. Moreover, being female and scoring higher on AdAS spectrum were statistically predictive factors for the presence of ON symptomatology. Among AdAS spectrum domains, higher scores on AdAS spectrum inflexibility and adherence to routine and restricted interests and rumination domains, as well as lower scores on verbal communication domain, were statistically predictive of orthorexic symptoms. CONCLUSIONS: Our findings highlight an overlap between ON and autism spectrum psychopathology. Further studies are needed to clarify the relationship between restrictive eating disorders and female autism phenotypes.

PTSD trajectories across different mental disorders in the second year of the COVID-19 pandemic in Italy: a naturalistic, longitudinal, multicenter study.

The potentially traumatic impact of the COVID-19 pandemic in subjects with pre-existing mental disorders is still unclear, especially regarding its long-term consequences. The aim of this study was to prospectively assess post-traumatic stress disorder (PTSD) and post-traumatic stress symptoms (PTSS) in patients with mental disorders, during the 3rd wave of the infection (T0, March-April 2021) while strict containment measures were applied in Italy, and after 3 months (T1, June-July 2021), with reduced restrictive measures. A total sample of 527 subjects, with different DSM-5 diagnoses, was consecutively enrolled at nine Italian psychiatric outpatient services. Assessments at T0 included: the Trauma and Loss Spectrum-Self Report (TALS-SR), the Impact of Event Scale-Revised (IES-R) and the Work and Social Adjustment Scale (WSAS). These two latter were repeated at T1. Results showed that at T0, 43.6% of the sample reported symptoms of PTSD, with females (p = .004), younger subjects (p = .011), unemployed/students (p = .011), and living with their parental families (p = .017), resulting more affected. Differences in PTSD rates emerged across diagnostic groups ranging from 10% in patients with psychoses up to 59% in those with feeding and eating disorders. An improvement at T1 emerged in all diagnostic groups for the IES-R scores, while WSAS scores improved only in subjects with mood disorders. In conclusions, subjects with mental disorders presented relevant rates of PTSD and PTSS at 1-year into the pandemic. Further long-term studies are needed to follow-up the course of pandemic traumatic burden especially in patients with severe mental disorders.

Naturalistic 6-month antidepressants follow-up in patients with fibromyalgia: impact on somatic and mood spectrum symptoms.

OBJECTIVES: Literature shows high rates of comorbidity between fibromyalgia (FM) and mood disorders, especially major depressive disorder (MMD), reported in more than half of the cases. Consistently, patients with FM also present high rates of mood spectrum symptoms, despite scant data are still available on the relationship with antidepressant treatment outcomes. The present study was aimed at exploring the clinical outcome of patients with FM-MDD comorbidity naturalistically treated with antidepressant drugs, besides the relationships between mood spectrum symptoms and the treatment response. METHODS: A total sample of 40 patients with FM and MDD, who started a treatment with an antidepressant drug, was recruited at the Rheumatology Unit of the University of Pisa, Italy. Patients were evaluated at baseline and after 1 (T1) and 6 months (T2) of the treatment with an antidepressant drug. Assessments included: the Mood Spectrum-Self Report (MOODS-SR) for mood spectrum symptoms, the Short Form Health Survey (SF-36) for the global functioning and the Clinical Global Impression (CGI) for the clinical severity and improvement. All instruments were administered at baseline and the SF-36 and CGI were repeated at T1 and T2. RESULTS: Twenty-eight (70%) patients reported an improvement at the CGI at T2. At T1 and T2 the CGI item-1 and most of the SF-36 domain scores significantly improved with respect to the T0, with the exception of the "role physical" and "role emotional" subscales. Improved patients reported higher scores in the energy depressive MOODS-SR domain. Furthermore, correlations emerged between several MOODS-SR domains and the CGI or SF-36 subscales scores at T0. CONCLUSIONS: Our results corroborate previous findings on the role of antidepressant drugs in the management not only of MDD symptoms, but also of the painful component of FM. FM patients should be investigated for Mood Spectrum symptomatology considering its prominent role on the manifestations of the disorder and treatment outcome.

Do somatic symptoms relate to PTSD and gender after earthquake exposure? A cross-sectional study on young adult survivors in Italy.

OBJECTIVE: Increasing evidence confirms a strict relationship between mental disorders and physical health. Particularly, stressful life events and post-traumatic stress disorder (PTSD) have been closely correlated with various physical disorders and somatic symptoms, such as chronic pain, gastrointestinal disorders, and headaches. The aim of this study was to investigate the emergence of somatic symptoms in a sample of young adult survivors 21 months after exposure to the L'Aquila 2009 earthquake, with particular attention to PTSD and gender impact. METHODS: Four hundred and fifty high-school senior students (253 male and 197 female) exposed to the 2009 L'Aquila earthquake, 21 months earlier, were enrolled and evaluated by the Trauma and Loss Spectrum Self-Report (TALS-SR), for symptomatological PTSD, and the Mood Spectrum Self-Report-Lifetime Version (MOODS-SR) "rhythmicity and vegetative functions" domain, for somatic symptoms. RESULTS: Significantly higher rates of endorsement of the MOODS-SR somatic symptoms emerged in survivors with PTSD compared to those without. Females reported higher rates of endorsement of at least one MOODS-SR somatic symptom compared to males; however, a Decision Tree model and a two-way analysis of variance model confirmed a significant effect of PTSD only. A multivariate logistical regression showed a significant association between the presence of at least one MOOD-SR somatic symptom and re-experiencing and maladaptive coping TALS-SR domains. CONCLUSION: This study corroborates a relevant impact of symptomatological PTSD, across both the genders, on somatic symptoms occurring in young adults after months from exposure to a massive earthquake.

A 3-Month Naturalistic Follow-Up Treatment With Selective Serotonin Reuptake Inhibitors in Frequent Attenders of General Medical Practice: What Correlates With a Good Response?

ABSTRACT: Frequent attenders (FAs), defined as patients repeatedly attending general practitioners, frequently exhibit underdiagnosed psychiatric comorbidities, leading to the hypothesis that frequent attendance may be related to an undetected psychiatric burden. This study explores the role of psychiatric comorbidities and psychopharmacological treatment on the clinical outcomes of a cohort of FAs of the general medical practice in Italy. The study included 75 FAs assessed by the Structured Clinical Interview for DSM-5, Clinical Global Impression, Global Assessment Functioning, and Illness Behavior Inventory, administered at baseline (T0) and after 3 months (T1). Data were analyzed on the bases of the presence of any mental disorder and selective serotonin reuptake inhibitor (SSRI) treatment, with respect to other psychopharmacological treatments. Results showed better outcomes among patients with a mental disorder, particularly anxiety, depression, and somatic symptoms disorders, and when under SSRI treatment. Our findings corroborate the role of psychiatric comorbidity on frequent attendance in the context of general clinical practice with a positive outcome when receiving appropriate treatment with SSRI.

Relationships Between Somatic Symptoms and Panic-Agoraphobic Spectrum Among Frequent Attenders of the General Practice in Italy.

Frequent attenders (FAs) of general practitioners (GPs) often complain of nonspecific physical symptoms that are difficult to define according to typical medical syndromes criteria but could be acknowledged as atypical manifestations of mental disorders. We investigated the possible correlation between somatic symptoms and panic-agoraphobic spectrum symptoms in a sample of 75 FAs of GPs in Italy, with particular attention to the impact on functional impairment. Assessments included the Patient Health Questionnaire, Panic-Agoraphobic Spectrum-Self-Report (PAS-SR) lifetime version, Global Assessment of Functioning, and Clinical Global Impression. The PAS-SR total and domains scores were significantly higher among low-functioning FAs, especially anxious somatizations, hypochondriasis, anxious expectation, and reassurance orientation domains, suggesting this undetected symptom may determine the selective attention to the physical symptoms, illness-phobic/hypochondriac elaboration, and GP frequent attendance, often aimed at searching for reassurance, leading to severe impact on overall functioning and often inefficacious treatments.

Synthesis, biological evaluation and molecular modeling of novel selective COX-2 inhibitors: sulfide, sulfoxide, and sulfone derivatives of 1,5-diarylpyrrol-3-substituted scaffold.

A novel series of 1,5-diarylpyrrol-3-sulfur derivatives (10-12) was synthesized and characterized by NMR and mass spectroscopy and x-ray diffraction. The biological activity of these compounds was evaluated in in vitro and in vivo tests to assess their COX-2 inhibitory activity along with anti-inflammatory and antinociceptive effect. Results showed that the bioisosteric transformation of previously reported alkoxyethyl ethers (9a-c) into the corresponding alkyl thioethers (10a-c) still leads to selective and active compounds being the COX-2 inhibitory activity for most of them in the low nanomolar range. The oxidation products of 10a,b were also investigated and both couple of sulfoxides (11a,b) and sulfones (12a,b) showed an appreciable COX-2 inhibitory activity. Molecular modeling studies were performed to investigate the binding mode of the representative compounds 10b, 11b, and 12b into COX-2 enzyme and to explore the potential site of metabolism of 10a and 10b due to the different in vivo efficacy. Among the developed compounds, compound 10b showed a significant in vivo anti-inflammatory and antinociceptive activity paving the way to develop novel anti-inflammatory drugs.

18F-VC701-PET and MRI in the in vivo neuroinflammation assessment of a mouse model of multiple sclerosis.

BACKGROUND: Positron emission tomography (PET) using translocator protein (TSPO) ligands has been used to detect neuroinflammatory processes in neurological disorders, including multiple sclerosis (MS). The aim of this study was to evaluate neuroinflammation in a mouse MS model (EAE) using TSPO-PET with 18F-VC701, in combination with magnetic resonance imaging (MRI). METHODS: MOG35-55/CFA and pertussis toxin protocol was used to induce EAE in C57BL/6 mice. Disease progression was monitored daily, whereas MRI evaluation was performed at 1, 2, and 4 weeks post-induction. Microglia activation was assessed in vivo by 18F-VC701 PET at the time of maximum disease score and validated by radioligand ex vivo distribution and immunohistochemistry at 2 and 4 weeks post-immunization. RESULTS: In vivo and ex vivo analyses show that 18F-VC701 significantly accumulates within the central nervous system (CNS), particularly in the cortex, striatum, hippocampus, cerebellum, and cervical spinal cord of EAE compared to control mice, at 2 weeks post-immunization. MRI confirmed the presence of focal brain lesions at 2 weeks post-immunization in both T1-weighted and T2 images. Of note, MRI abnormalities attenuated in later post-immunization phase. Neuropathological analysis confirmed the presence of microglial activation in EAE mice, consistent with the in vivo increase of 18F-VC701 uptake. CONCLUSION: Increase of 18F-VC701 uptake in EAE mice is strongly associated with the presence of microglia activation in the acute phase of the disease. The combined use of TSPO-PET and MRI provided complementary evidence on the ongoing disease process, thus representing an attractive new tool to investigate neuronal damage and neuroinflammation at preclinical levels.

Design, Synthesis, and Dynamics of a Green Fluorescent Protein Fluorophore Mimic with an Ultrafast Switching Function.

While rotary molecular switches based on neutral and cationic organic π-systems have been reported, structurally homologous anionic switches providing complementary properties have not been prepared so far. Here we report the design and preparation of a molecular switch mimicking the anionic p-HBDI chromophore of the green fluorescent protein. The investigation of the mechanism and dynamics of the E/Z switching function is carried out both computationally and experimentally. The data consistently support axial rotary motion occurring on a sub-picosecond time scale. Transient spectroscopy and trajectory simulations show that the nonadiabatic decay process occurs in the vicinity of a conical intersection (CInt) between a charge transfer state and a covalent/diradical state. Comparison of our anionic p-HBDI-like switch with the previously reported cationic N-alkyl indanylidene pyrrolinium switch mimicking visual pigments reveals that these similar systems translocate, upon vertical excitation, a similar net charge in the same axial direction.

Phenylindenone isomers as divergent modulators of p38α MAP kinase.

Two new fluorophenylindenone derivatives were designed as potential p38α MAPK modulators by preserving the key interactions of the vicinal pyridine/fluorophenyl pharmacophore with the enzyme protein. Interestingly, these two fluorophenylindenone isomers showed divergent activities, with compound 6 behaving as an inhibitor and 5 as a putative activator. These results were rationalized by docking studies and molecular dynamics simulations in terms of stabilization of DFG loop, by compound 5 in a conformation more accessible to phosphorylation.

Radiosynthesis and Preliminary Biological Evaluation of [18F]VC701, a Radioligand for Translocator Protein.

Positron emission tomography (PET) can be used to monitor in vivo translocator protein (TSPO) expression by using specific radioligands. Recently, several [11C]PK11195 analogues have been synthesized to improve binding stability and brain availability. [18F]VC701 was synthesized and validated in CD healthy rats by biodistribution and inhibition analysis. Imaging studies were also conducted on animals injected unilaterally in the striatum with quinolinic acid (QA) to evaluate the TSPO ligand uptake in a neuroinflammation/neurodegenerative model. [18F]VC701 was synthesized with a good chemical and radiochemical purity and specific activity higher than 37 GBq/µmol. Kinetic studies performed on healthy animals showed the highest tracer biodistribution in TSPO-rich organs, and preadministration of cold PK11195 caused an overall radioactivity reduction. Metabolism studies showed the absence of radiometabolites in the rat brain of QA lesioned rats, and biodistribution analysis revealed a progressive increase in radioactivity ratios (lesioned to nonlesioned striatum) during time, reaching an approximate value of 5 4 hours after tracer injection. These results encourage further evaluation of this TSPO radioligand in other models of central and peripheral diseases.

Supramolecular glycodendrimer-based hybrid drugs.

Specific noncovalent interactions are commonly used by nature to modulate numerous processes including cell recognition, viral adhesion, and transmembrane communications. Here we report on the design, synthesis, and preliminary characterization of new supramolecular glycodendrimer-based hybrid drugs based on adamantyl-modified glycodendrimers of third, fourth, or fifth generation (mPPI-G3-AdaB, mPPI-G4-AdaB, and mPPI-G5-AdaB) and a new heterobifunctional ligand. This component was tailored to bind through noncovalent interactions both the multimeric natural 5-HT3 receptor (through an optimized arylpiperazine pharmacophore) and the adamantyl groups located on the glycodendrimer surfaces (through a ß-cyclodextrin residue) giving rise to biorelevant supramolecular constructs.

Cross-Linked Hyaluronan Derivatives in the Delivery of Phycocyanin.

An easy and viable crosslinking technology, based on the "click-chemistry" reaction copper(I)-catalyzed azide-alkyne 1,3-dipolar cycloaddition (click-crosslinking), was applied to graft copolymers of medium molecular weight (i.e., 270 kDa) hyaluronic acid (HA) grafted with ferulic acid (FA) residues bearing clickable propargyl groups, as well as caffeic acid derivatives bearing azido-terminated oligo(ethylene glycol) side chains. The obtained crosslinked materials were characterized from the point of view of their structure and aggregation liability to form hydrogels in a water environment. The most promising materials showed interesting loading capability regarding the antioxidant agent phycocyanin (PC). Two novel materials complexes (namely HA(270)-FA-TEGEC-CL-20/PC and HA(270)-FA-HEGEC-CL-20/PC) were obtained with a drug-to-material ratio of 1:2 (w/w). Zeta potential measurements of the new complexes (-1.23 mV for HA(270)-FA-TEGEC-CL-20/PC and -1.73 mV for HA(270)-FA-HEGEC-CL-20/PC) showed alterations compared to the zeta potential values of the materials on their own, suggesting the achievement of drug-material interactions. According to the in vitro dissolution studies carried out in different conditions, novel drug delivery systems (DDSs) were obtained with a variety of characteristics depending on the desired route of administration and, consequently, on the pH of the surrounding environment, thanks to the complexation of phycocyanin with these two new crosslinked materials. Both complexes showed excellent potential for providing a controlled/prolonged release of the active pharmaceutical ingredient (API). They also increased the amount of drug that reach the target location, enabling pH-dependent release. Importantly, as demonstrated by the DPPH free radical scavenging assay, the complexation process, involving freezing and freeze-drying, showed no adverse effects on the antioxidant activity of phycocyanin. This activity was preserved in the two novel materials and followed a concentration-dependent pattern similar to pure PC.

Reactivity of a Morita-Baylis-Hillman adduct derivative bearing a triphenylamine moiety with lysine models.

The reactivity of Morita-Baylis-Hillman Adduct (MBHA) derivative 7 was studied with different primary amine derivatives such as n-butylamine, Na-acetyl-L-lysine methyl ester, and a poly-(L-lysine) derivative as lysine models to obtain information about the possible reactions in complex protein environments. MBHA derivative 7 reacted with n-butylamine or Na-acetyl-L-lysine methyl ester producing monoadducts 9a or 9c, which showed bright emission features in the green region at 526-535 nm with photoluminescence quantum yield values in solutions of 73% and 51%, respectively. Based on these results, MBHA derivative 7 can be considered an interesting new fluorogenic probe potentially useful in the labelling of basic amino acid residues. Furthermore, similar to other MBHA derivatives, compound 7 showed the tendency to produce diadducts especially in polar solvents system where specific interactions between the extended aromatic moieties may play a major role.

Combining spontaneous polymerization and click reactions for the synthesis of polymer brushes: a "grafting onto" approach.

Two novel benzofulvene monomers bearing propargyl or allyl groups have been synthesized by means of readily accessible reactions, and were found to polymerize spontaneously by solvent removal, in the apparent absence of catalysts or initiators, to give the corresponding polybenzofulvene derivatives bearing clickable propargyl or allyl moieties. The clickable propargyl and allyl groups were exploited in appropriate click reactions to develop a powerful and versatile "grafting onto" synthetic methodology for obtaining tailored polymer brushes.

[Grief: from the physiological reaction to the psychopathology.] / Lutto: dalla reazione fisiologica al disturbo da lutto prolungato.

The death of a loved one is a universal experience, and marker of the human condition. Grief, the cognitive, emotional, and behavioral responses to bereavement, is both a ubiquitous and a unique psychological process. Thus, health providers often find themselves in a dilemma, caught between the need to alleviate an individual's distress and impairment, and the danger of overly "pathologizing" their grief reaction. This chapter reviews how acute grief reactions generally evolve over time, the clinical presentation of complicated grief, and finally, other psychiatric disorders that might develop or be precipitated in the aftermath of the death of a loved one, particularly prolonged grief disorder.