Direct and indirect effects of psychological well-being and therapeutic alliance on therapy outcome in eating disorders.

Introduction: Outcome research in eating disorders (EDs) is commonly focused on psychopathological dysfunction. However, Ryff's model of psychological well-being (PWB) has shown promising-yet preliminary-results with ED patients. Additionally, despite substantial evidence highlighting the association between the therapeutic alliance and treatment outcome, findings in ED samples remain unclear. The present study aimed at exploring the direct effect of PWB dimensions and the early therapeutic alliance on ED patients' individual treatment responses, as well as the mediating role played by the early therapeutic alliance in the relationship between PWB dimensions and overall pre-post symptom change. Methods: A sample of N = 165 ED patients assigned female at birth, who were receiving treatment in a residential program, completed the Psychological Well-Being Scale at treatment intake and the Working Alliance Inventory after the first four psychotherapy sessions. Patients also completed the Outcome Questionnaire-45.2 at the same time point and during the week prior to discharge. Results: The PWB dimensions of autonomy, positive relations, and self-acceptance were associated with clinically significant change, while the dimensions of personal growth and self-acceptance were associated with reliable change. The early therapeutic alliance showed both direct and indirect effects on therapy outcome, predicting clinically significant and reliable symptom reduction. It also emerged as a significant mediator in the relationship between all PWB dimensions and overall symptomatic change. Conclusion: The identification of individual, adaptive characteristics in ED patients that might influence their development of an early therapeutic alliance may help therapists to predict relationship ruptures and tailor their interventions to enhance treatment effectiveness.

Clinical utility of the at-risk for psychosis state beyond transition: A multidimensional network analysis.

To be relevant to healthcare systems, the clinical high risk for psychosis (CHR-P) concept should denote a specific (i.e., unique) clinical population and provide useful information to guide the choice of intervention. The current study applied network analyses to examine the clinical specificities of CHR-P youths compared to general help-seekers and non-CHR-P youth. 146 CHR-P (mean age = 14.32 years) and 103 non-CHR-P (mean age = 12.58 years) help-seeking youth were recruited from a neuropsychiatric unit and assessed using the Structured Interview for Psychosis-Risk Syndromes, Children's Depression Inventory, Multidimensional Anxiety Scale for Children, Global Functioning: Social, Global Functioning: Role, and Wechsler Intelligence Scale for Children/Wechsler Adult Intelligence Scale. The first network structure comprised the entire help-seeking sample (i.e., help-seekers network), the second only CHR-P patients (i.e., CHR-P network), and the third only non-CHR-P patients (i.e., non-CHR-P network). In the help-seekers network, each variable presented at least one edge. In the CHR-P network, two isolated "archipelagos of symptoms" were identified: (a) a subgraph including functioning, anxiety, depressive, negative, disorganization, and general symptoms; and (b) a subgraph including positive symptoms and the intelligence quotient. In the non-CHR-P network, positive symptoms were negatively connected to functioning, disorganization, and negative symptoms. Positive symptoms were less connected in the CHR-P network, indicating a need for specific interventions alongside those treating comorbid disorders. The findings suggest specific clinical characteristics of CHR-P youth to guide the development of tailored interventions, thereby supporting the clinical utility of the CHR-P concept.

Problematic internet use in emerging adulthood to escape from maternal helicopter parenting: defensive functioning as a mediating mechanism.

The increasing use of the Internet has raised concerns about its problematic use, particularly among emerging adults who grew up in a highly digitalized world. Helicopter parenting, characterized by excessive involvement, overcontrol, and developmentally inappropriate behavior, has been identified as a potential factor contributing to problematic Internet use (PIU). Under these circumstances, considering that emerging adults navigate their adult lives and strive to reduce their sense of being in-between, implicit emotion regulation strategies, such as defense mechanisms, may help comprehend PIU. The present questionnaire-based study investigated the associations between maternal and paternal helicopter parenting and PIU through defensive functioning among a community sample of 401 cisgender emerging adults (71.82% females; 82.04% heterosexuals; Mage=24.85, SD=2.52) living in Italy. About one-fourth (25.19%) reported PIU. Greater maternal, B=0.904, SE=0.235, p<.001, but not paternal, B=0.343, SE=0.188, p=.068, helicopter parenting was significantly associated with PIU through a less mature defensive functioning. Conversely, neither maternal, B=1.158, SE=0.722, p=.109, nor paternal, B=0.355, SE=0.731, p=.628, helicopter parenting had a direct association with PIU. The results suggest the importance for psychotherapists to incorporate individuals' defense mechanisms and parent-child relationship history when designing tailored interventions for effective treatment of PIU. This emphasis is crucial because, in the context of a developmentally appropriate parenting style, relying on more mature defenses after psychotherapeutic intervention can lead to healthier adjustment among emerging adults.

Body Uneasiness and Dissatisfaction Among Lesbian, Gay, Bisexual, and Heterosexual Persons.

Introduction: While sexual minority people have been widely considered at risk for developing a range of body image concerns, evidence of body dissatisfaction and shame amongst LGB (lesbian, gay, and bisexual) individuals is mixed. This study investigated differences in body uneasiness, body dissatisfaction, and self-blaming/attacking attitudes between LGB and heterosexual individuals, as well as within LGB groups, while also examining the predictive role of body mass index (BMI). Methods: A sample of cisgender lesbian women (n = 163), gay men (n = 277), bisexual women (n = 135), bisexual men (n = 39), heterosexual women (n = 398), and heterosexual men (n = 219) completed an online survey assessing different aspects of body image between May and July 2020. Results: Gay and bisexual men reported greater body image disturbance and self-blaming attitudes relative to heterosexual men. In contrast, lesbian women reported lower body uneasiness than their bisexual and heterosexual counterparts, but greater self-hate. Moreover, lesbian and bisexual women showed more body dissatisfaction than gay men, and bisexual individuals reported more body uneasiness than individuals in other sexual minority subgroups. Higher BMI emerged as a significant predictor of body image concerns and dissatisfaction. Conclusions: Body image dimensions showed sexual identity-based differences. Determining the specific nuances of body image in LGB individuals can provide important information on potential risk factors that may impact mental health outcomes. Policy Implications: In-depth knowledge of body dissatisfaction and uneasiness in individuals with LGB identities may have critical implications for the development of personalized prevention and treatment strategies.

From symptoms to subjective and bodily experiences: the contribution of the Psychodynamic Diagnostic Manual (PDM-2) to diagnosis and treatment monitoring in eating disorders.

PURPOSE: Atheoretical and descriptive conceptualizations of eating disorders (EDs) have faced substantial criticism due to their limited ability to assess patients' subjective characteristics and experiences, as needed to determine the most appropriate treatment options. The present article provides an overview of the clinical and empirical literature supporting the potential contribution of the Psychodynamic Diagnostic Manual (PDM-2) to both diagnostic assessment and treatment monitoring. METHODS: Following a discussion of the most relevant shortcomings of current diagnostic models of EDs and a description of the rationale and structure of the PDM-2, evidence supporting the core PDM-2 dimensions of ED patients' subjective experiences (i.e., affective states, cognitive processes, relational patterns, somatic/bodily experiences and states) are examined, alongside their relevance to ED diagnosis and treatment. RESULTS: Overall, the reviewed studies support the diagnostic importance of these patterns of subjective experiences in EDs, highlighting their potential role as either predisposing or maintaining factors to target in psychotherapy. A growing body of multidisciplinary evidence also shows that bodily and somatic experiences are central to the diagnosis and clinical management of ED patients. Moreover, there is evidence that a PDM-based assessment may enable closer monitoring of patient progress during treatment, with regard to both subjective experiences and symptom patterns. CONCLUSIONS: The study suggests that current diagnostic frameworks for EDs would benefit from the addition of a person-centered perspective that considers not only symptoms, but also patients' full range of functioning-including their deep and surface-level emotional, cognitive, interpersonal, and social patterns-to improve patient-tailored interventions. LEVEL OF EVIDENCE: Level V, narrative review.

The Mediator Effect of Personality on the Relationship Between Symptomatic Impairment and Treatment Outcome in Eating Disorders.

Features of personality disorders (PDs) have been found to explain meaningful variance in the onset, maintenance, and symptomatic presentation of eating disorders (EDs), and a co-occurent personality pathology is commonly associated with poorer response to ED treatment. The "pathoplasty model" of the relationship between personality and EDs implies that, once both conditions are established, they are likely to interact in ways that modify therapy outcome; however, to date, no studies have explored overall personality functioning, and especially PD clusters, as a mediator of treatment outcome. The present study aimed at conjointly exploring the associations between personality functioning and PDs, respectively, with pre-treatment ED symptomatic impairment and therapy outcome; and the mediating role of personality variables. At treatment onset, a sample of 107 women with ED problems were evaluated using both the Structured Clinical Interview for DSM-5 (SCID-5-CV) and the Shedler-Westen Assessment Procedure-200 (SWAP-200)-a clinician-rated procedure to dimensionally assess personality. Participants were also asked to complete self-report questionnaires on overall ED symptomatology, symptoms of binge eating and purging behaviors, and therapy outcome. The findings showed that, over and above the categorical ED diagnosis, the SWAP-200 healthy personality functioning score mediated the relationship between baseline ED symptom severity and therapy outcome, as well as the association between baseline bulimic symptoms and treatment outcome; furthermore, SWAP-200 Cluster B PD scores mediated the link between baseline binge eating and purging symptoms and therapy outcome, whereas scores in Clusters A and C showed no significant effects. The findings suggest that personality-based outcome research may improve treatment effectiveness in this difficult-to-treat population.

Affective temperaments and depressive symptoms: The mediating role of attachment.

BACKGROUND: Studies showed that affective temperaments and attachment are associated with depressive symptoms, and that they bi-directionally influence each-other. The aim of this study is to explore mechanisms underlying the relationship between the affective temperaments (i.e., depressive, cyclothymic, irritable, hyperthymic and anxious), interview-based attachment, and depressive symptoms. METHODS: A sample of 61 adolescents and young adults outpatients were asked to complete the Temperament Evaluation of Memphis, Pisa, and San Diego Autoquestionnaire (TEMPS-A) and the Hamilton Rating Scale for Depression (HAM-D). Attachment was assessed through the Adult Attachment Interview (AAI) and employing a dimensional approach to obtain continuous measures. Mediation models were performed with the affective temperaments as predictor, depressive symptoms as the criterion variable, and attachment dimensions as mediators. RESULTS: Findings showed significant direct effects between all the affective temperaments and depressive symptoms. Only the cyclothymic (ß = 0.22; SE = 0.1; 95% IC = 0.05, 0.42) and irritable (ß = 0.21; SE = 0.09; 95% IC = 0.04, 0.4) temperaments showed an indirect effect on depressive symptoms through secure-insecure attachment. Dismissing attachment did not predict either the affective temperaments nor depressive symptoms. Preoccupied attachment significantly predicts depressive symptoms and, when controlling for it, the hyperthymic temperament no longer directly associates with depressive symptoms. LIMITATIONS: The cross-sectional study design limit conclusion about causation and directionality. CONCLUSIONS: Secure attachment could be a protective factor for depressive symptoms for individuals with a cyclothymic or irritable temperament. Differently, the hyperthymic temperament loses its protective role toward depressive symptoms when accounting for preoccupied attachment.

Personality as a predictor of symptomatic change in a residential treatment setting for anorexia nervosa and bulimia nervosa.

PURPOSE: Although personality has been widely researched in patients with anorexia nervosa (AN) and bulimia nervosa (BN), the nature of this relationship has not yet been clearly articulated. The pathoplasty model theorizes that personality might shape symptomatic presentation and thus affect therapeutic outcomes, but more research is needed. The present study aimed at investigating the predictive value of a broad spectrum of personality traits in determining AN and BN treatment outcomes, considering both the statistical and clinical significance of the therapeutic change. METHODS: Eighty-four female patients with AN and BN treated in a residential program were evaluated at treatment onset using the Shedler-Westen Assessment Procedure-200-a clinician-rated measure of personality disorders and healthy personality functioning. At both intake and discharge, patients completed the Eating Disorder Inventory-3 to assess eating symptoms and the Outcome Questionnaire-45.2 to evaluate overall impairment. RESULTS: Considering overall ED symptomatic change, multiple regression analyses showed that, even when controlling for baseline symptoms and DSM-5 categories, schizoid (B = 0.41, p ≤ 0.01), avoidant (B = 0.31, p ≤ 0.05), and paranoid (B = 0.25, p ≤ 0.05) personality features predicted worse therapeutic outcomes. Similar results were found when applying the clinical significance approach, with the emotionally dysregulated factor as an additional negative predictor of significant/reliable change (B = - 0.09; p < 0.01). Healthy personality functioning predicted better therapeutic outcomes (B = - 0.34, p ≤ 0.001). CONCLUSIONS: Pathoplastic models and personality-based research in this clinical population have the potential to inform effective treatment strategies by targeting relevant individual factors. LEVEL OF EVIDENCE: Level III, longitudinal cohort study.

How to Improve Compliance with Protective Health Measures during the COVID-19 Outbreak: Testing a Moderated Mediation Model and Machine Learning Algorithms.

In the wake of the sudden spread of COVID-19, a large amount of the Italian population practiced incongruous behaviors with the protective health measures. The present study aimed at examining psychological and psychosocial variables that could predict behavioral compliance. An online survey was administered from 18-22 March 2020 to 2766 participants. Paired sample t-tests were run to compare efficacy perception with behavioral compliance. Mediation and moderated mediation models were constructed to explore the association between perceived efficacy and compliance, mediated by self-efficacy and moderated by risk perception and civic attitudes. Machine learning algorithms were trained to predict which individuals would be more likely to comply with protective measures. Results indicated significantly lower scores in behavioral compliance than efficacy perception. Risk perception and civic attitudes as moderators rendered the mediating effect of self-efficacy insignificant. Perceived efficacy on the adoption of recommended behaviors varied in accordance with risk perception and civic engagement. The 14 collected variables, entered as predictors in machine learning models, produced an ROC area in the range of 0.82-0.91 classifying individuals as high versus low compliance. Overall, these findings could be helpful in guiding age-tailored information/advertising campaigns in countries affected by COVID-19 and directing further research on behavioral compliance.

How to be a secure base: therapists' attachment representations and their link to attunement in psychotherapy.

This paper investigates if therapists' attachment classifications are linked to different types of therapists' attunement in session. We present coding procedures and validation of the Therapist Attunement Scales (TASc), a transcript-based instrument that assesses attunement in therapy and that we hypothesized would be associated with therapists' attachment classifications. Fifty therapists were administered the Adult Attachment Interview (AAI), and therapy sessions with each therapist were rated with the TASc. Therapists also completed the WAI-T to assess the therapeutic alliance and divergent validity of the TASc. Results indicate strong inter-rater reliability and stability across consecutive sessions of the TASc, as well as high convergent validity between one session rated with the TASc and the AAI rated independently (κ = .81). No significant associations were found with the WAI-T, which offers preliminary evidence of divergent validity of the TASc. These results suggest that therapists of different attachment classifications may attune to patients in distinct ways. These results also present the TASc as a valid measure of therapists' attachment in psychotherapy and as a promising tool for training and supervision.

Evaluating empirically valid and clinically meaningful change in intensive residential treatment for severe eating disorders at discharge and at a 6-month follow-up.

PURPOSE: The present study evaluated the statistical and clinical significance of symptomatic change at discharge and after 6 months of an intensive residential treatment for patients with eating disorders (ED), and explored the individual factors that may affect therapeutic outcomes. METHODS: A sample of 118 female ED patients were assessed at intake and discharge on the following dimensions: BMI, ED-specific symptoms, depressive features, and overall symptomatic distress. A subsample of 59 patients filled out the same questionnaires at a 6-month follow-up after discharge. RESULTS: Findings evidenced statistically significant changes in all outcome measures at both discharge and follow-up. Between 30.1 and 38.6% of patients at discharge and 35.2-54.2% at the 6-month follow-up showed clinically significant symptomatic change; additionally, 19.8-29.1% of patients at discharge and 22.9-38.3% at follow-up improved reliably. However, 34.9-39.8% remained unchanged and 2-4.8% worsened. At the 6-month follow-up, 21.3-25.9% showed no symptomatic change and 0-3.7% had deteriorated. Unchanged and deteriorated patients had an earlier age of ED onset and were more likely to suffer a comorbid personality pathology and to be following concurrent pharmacological treatment. CONCLUSIONS: Results suggested that intensive and multimodal residential treatment may be effective for the majority of ED patients, and that therapeutic outcomes tend to improve over time. Prevention strategies should focus on early onset subjects and those with concurrent personality pathology. LEVEL OF EVIDENCE: Level III, evidence obtained from a longitudinal cohort study.

Do therapists' subjective variables impact on psychodynamic psychotherapy outcomes? A systematic literature review.

BACKGROUND: Despite growing attention to the general therapist effects in a wide range of clinical settings, little is known about the individual, cross-situational, and therapy-nonspecific variables that impact on the differential effectiveness of clinicians. The current study is a systematic review of the evidence relating to the influence of therapist's subjective characteristics on outcomes of psychodynamic psychotherapies. METHOD: A multistage and systematic search of articles published between 1987 and 2017 identified 30 relevant studies, which were organized into 6 areas according to the specific therapist's variable considered. RESULTS: Therapists' interpersonal functioning and skills showed the strongest evidence of a direct effect on treatment outcomes. Furthermore, there were preliminary evidence that therapists' attachment styles, their interpersonal history with caregivers, and their self-concept might affect outcomes through interaction effects with other constructs, such as technical interventions, patient's pathology, and therapeutic alliance. The high variability between studies on therapists' overall reflective or introspective abilities and personality characteristics suggested the need for more systematic research in these areas, whereas therapists' values and attitudes showed small effects on therapeutic outcome. CONCLUSIONS: The present review clarifies how a deep examination of the contribution of therapists' subjective characteristics can help elucidate the complex association between relational and technical factors related to the outcome of psychodynamic treatments.

Countertransference when working with narcissistic personality disorder: An empirical investigation.

Narcissistic personality disorder (NPD) is one of the most challenging clinical syndromes to treat in psychotherapy, especially due to the difficulties of establishing a good enough therapist-patient relationship. Countertransference responses to NPD can be particularly intense, frustrating, and difficult to manage, as is often reported in the clinical literature though not clearly supported empirically. The aims of this study were to (a) investigate the relationship between patients' NPD and therapists' responses; (b) examine the associations between patient, clinician, therapy variables and clinicians' reactions during treatment of NPD patients; and (c) provide an empirically derived portrait of countertransference with NPD. A sample of psychiatrists and clinical psychologists (N = 67) completed the Therapist Response Questionnaire to identify patterns of countertransference, the Shedler-Westen Assessment Procedure-200, and the Global Assessment of Functioning Scale to assess the personality pathology and psychosocial functioning of a patient in their care. The results showed that NPD was positively associated with hostile/angry, criticized/devalued, helpless/inadequate, and disengaged countertransference and negatively associated with therapists' positive response, regardless of patients' personality and psychosocial functioning. NPD patients with stronger traits of cluster B personality pathology tended to elicit more negative and heterogeneous countertransference reactions than NPD patients without these features. The countertransference patterns with NPD patients were not strongly influenced by the variables of clinicians and therapy, with the exception of clinical experience. Overall, the portrait of therapists' reactions to NPD provided a clinically nuanced and empirically founded description strongly resembling theoretical-clinical accounts. The therapeutic implications of these findings were discussed. (PsycINFO Database Record

Does reflective functioning mediate the relationship between attachment and personality?

Mentalization, operationalized as reflective functioning (RF), can play a crucial role in the psychological mechanisms underlying personality functioning. This study aimed to: (a) study the association between RF, personality disorders (cluster level) and functioning; (b) investigate whether RF and personality functioning are influenced by (secure vs. insecure) attachment; and (c) explore the potential mediating effect of RF on the relationship between attachment and personality functioning. The Shedler-Westen Assessment Procedure (SWAP-200) was used to assess personality disorders and levels of psychological functioning in a clinical sample (N = 88). Attachment and RF were evaluated with the Adult Attachment Interview (AAI) and Reflective Functioning Scale (RFS). Findings showed that RF had significant negative associations with cluster A and B personality disorders, and a significant positive association with psychological functioning. Moreover, levels of RF and personality functioning were influenced by attachment patterns. Finally, RF completely mediated the relationship between (secure/insecure) attachment and adaptive psychological features, and thus accounted for differences in overall personality functioning. Lack of mentalization seemed strongly associated with vulnerabilities in personality functioning, especially in patients with cluster A and B personality disorders. These findings provide support for the development of therapeutic interventions to improve patients' RF.

Qualification of HSQC methods for quantitative composition of heparin and low molecular weight heparins.

An NMR HSQC method has recently been proposed for the quantitative determination of the mono- and disaccharide subunits of heparin and low molecular weight heparins (LMWH). The focus of the current study was the validation of this procedure to make the 2D-NMR method suitable for pharmaceutical quality control applications. Pre-validation work investigated the effects of several experimental parameters to assess robustness and to optimize critical factors. Important experimental parameters were pulse sequence selection, equilibration interval between pulse trains and temperature. These observations were needed so that the NMR method was sufficiently understood to enable continuous improvement. A standard validation study on heparin then examined linearity, repeatability, intermediate precision and limits of detection and quantitation; selected validation parameters were also determined for LMWH.

The Contribution of Sidney Blatt's Two-Polarities Model to the Psychodynamic Diagnostic Manual.

This article reviews the theoretical and empirical contributions of Blatt's two-polarities model of personality development and psychopathology to the second edition of the Psychodynamic Diagnostic Manual (PDM-2). First, we briefly provide an overview of the manual's main features and describe the guiding principles of the revision process. We then discuss in more detail how Blatt's model, which is focused on the dialectic interaction between anaclitic-introjective and relatedness vs self-definition dimensions in both normal and disrupted personality development, increases the PDM-2's theoretical and empirical comprehensiveness and clinical utility, especially concerning the classification and assessment of personality and overall mental functioning in adult populations. Finally, we address the implications for the therapeutic process, showing how anaclitic and introjective patients may be differentially responsive to different psychodynamic techniques (e.g., supportive or expressive interventions). Taken together, these considerations demonstrate the importance of a more theory-driven and empirically informed diagnostic system that embraces, in accordance with Sidney Blatt's empirically supported and psychoanalytically-oriented ideas, the complexities of human experience (both normal and pathological) and captures the subjective and underlying dynamics of psychological symptoms and syndromes.

Interactions and effects of BSA-functionalized single-walled carbon nanotubes on different cell lines.

Functionalized carbon nanotubes (CNTs) have shown great promise in several biomedical contexts, spanning from drug delivery to tissue regeneration. Thanks to their unique size-related properties, single-walled CNTs (SWCNTs) are particularly interesting in these fields. However, their use in nanomedicine requires a clear demonstration of their safety in terms of tissue damage, toxicity and pro-inflammatory response. Thus, a better understanding of the cytotoxicity mechanisms, the cellular interactions and the effects that these materials have on cell survival and on biological membranes is an important first step for an appropriate assessment of their biocompatibility. In this study we show how bovine serum albumin (BSA) is able to generate homogeneous and stable dispersions of SWCNTs (BSA-CNTs), suggesting their possible use in the biomedical field. On the other hand, this study wishes to shed more light on the impact and the interactions of protein-stabilized SWCNTs with two different cell types exploiting multidisciplinary techniques. We show that BSA-CNTs are efficiently taken up by cells. We also attempt to describe the effect that the interaction with cells has on the dielectric characteristics of the plasma membrane and ion flux using electrorotation. We then focus on the BSA-CNTs' acute toxicity using different cellular models. The novel aspect of this work is the evaluation of the membrane alterations that have been poorly investigated to date.

New macrocyclic analogs of the natural histone deacetylase inhibitor FK228; design, synthesis and preliminary biological evaluation.

Among the natural histone deacetylase inhibitors (HDACi), the bicyclic depsipeptide macrolactone FK228 stands out for its unique chemical structure and mechanism of action. In order to expand the chemical diversity, exploiting the FK228 peculiar structure, we have synthesized a collection of 24 simplified novel analogs. A first series consists of bicyclic macrolactones, where the carboxy terminus of the natural compound was substituted by peptidomimetic aminomethylphenylacetic acid derivatives. These analogs, 7a-i, showed submicromolar cytotoxic activity, even though very low inhibitory activity against HDAC enzymes, suggesting that most probably they behave with a mechanism different from the natural compound. One of the most active members in the group, 7g, was evaluated in vivo and exhibited significant antitumor activity. This evidence supports that the activity is unrelated to HDAC inhibition and these compounds represent a novel series of promising active agents. Another analog series consists of monocyclic macrolactones, 9a-c and 10a-d which lack the disulfide bridge and bear the protected sulfur on the linear external chain; they showed similar cytotoxic activities compared to the natural compound, but proved to be very sensitive to the nature of the sulfur protection. In fact, when the sulfur was protected by an 1-octanoyl residue, like in 9b, the product displayed a one digit nanomolar activity. The results provide evidence that our approach may be followed to develop novel series of FK228 analogs.

Diameter-dependent release of a cisplatin pro-drug from small and large functionalized carbon nanotubes.

The use of platinum-based chemotherapeutic drugs in cancer therapy still suffers from severe disadvantages, such as lack of appropriate selectivity for tumor tissues and insurgence of multi-drug resistance. Moreover, drug efficacy can be attenuated by several mechanisms such as premature drug inactivation, reduced drug uptake inside cells and increased drug efflux once internalized. The use of functionalized carbon nanotubes (CNTs) as chemotherapeutic drug delivery systems is a promising strategy to overcome such limitations due to their ability to enhance cellular internalization of poorly permeable drugs and thus increase the drug bioavailability at the diseased site, compared to the free drug. Furthermore, the possibility to encapsulate agents in the nanotubes' inner cavity can protect the drug from early inactivation and their external functionalizable surface is useful for selective targeting. In this study, a hydrophobic platinum(IV) complex was encapsulated within the inner space of two different diameter functionalized multi-walled CNTs (Pt(IV)@CNTs). The behavior of the complexes, compared to the free drug, was investigated on both HeLa human cancer cells and RAW 264.7 murine macrophages. Both CNT samples efficiently induced cell death in HeLa cancer cells 72 hours after the end of exposure to CNTs. Although the larger diameter CNTs were more cytotoxic on HeLa cells compared to both the free drug and the smaller diameter nanotubes, the latter allowed a prolonged release of the encapsulated drug, thus increasing its anticancer efficacy. In contrast, both Pt(IV)@CNT constructs were poorly cytotoxic on macrophages and induced negligible cell activation and no pro-inflammatory cytokine production. Both CNT samples were efficiently internalized by the two types of cells, as demonstrated by transmission electron microscopy observations and flow cytometry analysis. Finally, the platinum levels found in the cells after Pt(IV)@CNT exposure demonstrate that they can promote drug accumulation inside cells in comparison with treatment with the free complex. To conclude, our study shows that CNTs are promising nanocarriers to improve the accumulation of a chemotherapeutic drug and its slow release inside tumor cells, by tuning the CNT diameter, without inducing a high inflammatory response.

Unravelling structural information from complex mixtures utilizing correlation spectroscopy applied to HSQC spectra.

The first use of statistical correlation spectroscopy to extract chemical information from 2D-HSQC spectra, termed HSQC correlation spectroscopy (HSQCcos), is reported. HSQCcos is illustrated using heparin, a heterogeneous polysaccharide, whose diverse composition causes signals in HSQC spectra to disperse. HSQCcos has been used to probe the chain modifications that cause this effect and reveals hitherto unreported structural details. An interesting finding was that the signal for position 2 of trisulfated glucosamine [N-, 3-O-, and 6-O-sulfated] (A*) is bifurcated, owing to the presence of A* residues in both the "normal" antithrombin binding site and also at the nonreducing end of the molecule, which is reported in intact heparin for the first time. The method was also applied to investigating the environment around other rare sequences/disaccharides, suggesting that the disaccharide; 2-O-sulfated iduronic acid linked to 6-O-sulfated N-glucosamine, which contains a free amine at position 2, is adjacent to the heparin linkage region. HSQCcos can extract chemically related signals from information-rich spectra obtained from complex mixtures such as heparin.