Native extracellular matrix probes to target patient- and tissue-specific cell-microenvironment interactions by force spectroscopy.

Atomic Force Microscopy (AFM) is successfully used for the quantitative investigation of the cellular mechanosensing of the microenvironment. To this purpose, several force spectroscopy approaches aim at measuring the adhesive forces between two living cells and also between a cell and an appropriate reproduction of the extracellular matrix (ECM), typically exploiting tips suitably functionalised with single components (e.g. collagen, fibronectin) of the ECM. However, these probes only poorly reproduce the complexity of the native cellular microenvironment and consequently of the biological interactions. We developed a novel approach to produce AFM probes that faithfully retain the structural and biochemical complexity of the ECM; this was achieved by attaching to an AFM cantilever a micrometric slice of native decellularised ECM, which was cut by laser microdissection. We demonstrate that these probes preserve the morphological, mechanical, and chemical heterogeneity of the ECM. Native ECM probes can be used in force spectroscopy experiments aimed at targeting cell-microenvironment interactions. Here, we demonstrate the feasibility of dissecting mechanotransductive cell-ECM interactions in the 10 pN range. As proof-of-principle, we tested a rat bladder ECM probe against the AY-27 rat bladder cancer cell line. On the one hand, we obtained reproducible results using different probes derived from the same ECM regions; on the other hand, we detected differences in the adhesion patterns of distinct bladder ECM regions (submucosa, detrusor, and adventitia), in line with the disparities in composition and biophysical properties of these ECM regions. Our results demonstrate that native ECM probes, produced from patient-specific regions of organs and tissues, can be used to investigate cell-microenvironment interactions and early mechanotransductive processes by force spectroscopy. This opens new possibilities in the field of personalised medicine.

Psychopharmacological treatment in borderline personality disorder: A pilot observational study in a real-world setting.

Psychotherapy is the cornerstone of treatment for borderline personality disorder (BPD) while pharmacotherapy should be considered only as an adjunctive intervention. In clinical practice, however, most of BPD patients only receive medication. The aim of the study is to first describe pharmacological treatment in BPD patients in Italy and secondly to evaluate if comorbidity or illness severity are associated with the prescription of different class compounds. Data on pharmacological treatment and clinical evaluation of 75 BPD patients were collected in 5 clinical settings. The association between comorbidity and medication was assessed. Moreover, we evaluated the association between pharmacotherapy and severity, defined by a cluster analysis aimed at detecting different groups of patients. Most of the participants (82.7%) were characterized by polypharmacy, with a mean of 2.4 medications per person. Interestingly, the prescription didn't seem to depend on/be based on the severity of the disorder and was only partially determined by the presence of comorbidity. In conclusion, our findings are similar to what described in other clinical studies, supporting the idea that medication management for BPD is only partially coherent with international guidelines. This pilot study confirms the need for more rigorous studies to gain greater understanding of this topic and diminish the gap between guidelines and the real clinical world.

Clinical predictors of drug response in obsessive-compulsive disorder.

The aim of this study was to evaluate which clinical variables might influence the antiobsessional response to proserotonergic drugs in a sample of patients with obsessive-compulsive disorder (OCD). One hundred fifty-nine patients with DSM-IV OCD underwent a 12-week standardized treatment with fluvoxamine, clomipramine, citalopram, or paroxetine. According to treatment response, defined as a reduction of the Yale-Brown Obsessive Compulsive Scale total score >35%, patients were divided into two groups. Ninety patients (56.6%) responded to treatment and 69 (43.4%) did not. Responders had a significantly higher frequency of positive family history for OCD (FH-OCD) in their first-degree relatives, whereas nonresponders had an earlier onset and a higher frequency of "poor insight" subtype and somatic obsessions. The predictive value of all these variables was tested by a stepwise logistic regression analysis that confirmed poor insight and FH-OCD to be the best predictors of poor and good drug treatment response, respectively. These preliminary findings need additional investigations toward a better definition of the genetic and biological heterogeneity of patients with OCD, and they underlie the importance of collecting the insight score and family history for psychiatric disorders in the pretreatment assessment.

Relationship between obsessive-compulsive personality disorder and obsessive-compulsive disorder.

This study investigated the presence of obsessive-compulsive personality disorder (OCPD) in a group of 277 patients (88 with obsessive-compulsive disorder [OCD], 58 with major depressive disorder [MDD], and 131 with panic disorder [Panic]) to test the specificity of the relationship between OCPD and OCD. OCPD is statistically significantly more frequent in patients with OCD than in those with Panic and MDD. The distribution of single criteria of OCPD in the three groups does not differ significantly. Discriminant analysis selects a list of items that provide a correct classification rate of 66% based on OCPD criteria selected by canonical function. OCD patients with and without OCPD do not differ in sex, age of onset, duration of illness, positive family history for Tics disorder/Tourette syndrome (TS), or morbidity risk for OCD.

Tryptophan depletion in obsessive-compulsive patients.

Twelve patients with obsessive-compulsive disorder were studied after the administration of a mixture of amino acids devoid of tryptophan (TRP) or a mixture containing all the essential amino acids, in a double-blind, crossover design. The TRP-free mixture caused a marked depletion of plasma TRP. After TRP decrease, mean ratings of obsessions and compulsions, measured by Visual Analogue Scales (VAS) ratings, did not worsen. In contrast with other reports in literature, TRP depletion also failed to alter mood in our subjects.

A molecular investigation suggests no relationship between obsessive-compulsive disorder and the dopamine D2 receptor.

Some studies suggest a relationship between obsessive-compulsive disorder (OCD) and Gilles de la Tourette's syndrome. The pathophysiology of the latter may involve the dopamine system. We screened three important exons of the dopamine D2 receptor (DRD2) gene for mutations in a group of OCD patients with or without tics. No structural changes were found, suggesting no relationship between DRD2 and OCD. Moreover, the frequency of the polymorphism in exon 6 was different from that found in schizophrenics.

Psychiatric disorders in the families of patients with obsessive-compulsive disorder.

The rate of comorbid diagnoses in a group of 92 patients with obsessive-compulsive disorder (OCD) was examined, with particular attention being paid to mood disorders. The family history method was used to study the frequency of psychiatric disorders in the patients' families and to analyze the characteristics of the familial loading for OCD and mood disorders. A comorbid diagnosis of mood disorder occurred in 35.9% of the patients. The morbidity risk for OCD in the patients' families accounted for 3.4%; when 21 patients with an age of onset under 14 were examined, the morbidity risk in first degree relatives reached 8.8%. This tendency did not appear to be true for mood disorders.

Psychoimmunoendocrine aspects of panic disorder.

Immunological, neuroendocrine and psychological parameters were examined in 14 psychophysically healthy subjects and in 17 panic disorder patients before and after a 30-day course of alprazolam therapy. T lymphocyte proliferation in response to the mitogen phytohemagglutinin, lymphocyte beta-endorphin (beta-EP) concentrations, plasma ACTH, cortisol and beta-EP levels were examined in basal conditions and after corticotropin-releasing hormone (CRH) stimulation. Cortisol inhibition by dexamethasone (DST) and basal growth hormone (GH) and prolactin levels were also examined. Depression, state or trait anxiety, anticipatory anxiety, agoraphobia, simple and social phobias, severity and frequency of panic attacks were monitored by rating scales. The immune study did not reveal any significant difference between patients and controls, or any effect of alprazolam therapy. The hormonal data for the two groups were similar, except for higher than normal basal ACTH and GH plasma levels, lower than normal ratios between the ACTH and cortisol responses to CRH, and blunted DST in some patients. All the impairments improved after alprazolam therapy, in parallel with decreases in anxiety and in severity and frequency of panic attacks.

DSM-III-R personality disorders in panic and obsessive-compulsive disorder: a comparison study.

Forty-eight patients with panic disorder/agoraphobia (PAD) and 30 with obsessive-compulsive disorder (OCD) were assessed for DSM-III-R axis II personality disorders (PD) and the presence of the same anxiety disorder in the relatives of probands (homotypic disorders). No specific personality disorder was present significantly more often in either of the two groups. Agoraphobia was not associated with higher rates of axis II disorders in PAD patients. Duration of illness did not influence the presence of a PD in patients of both groups. Secondary cases of the same anxiety disorder were significantly more common among first-degree relatives of PAD patients. A discriminant analysis performed on the most frequent personality traits of both groups provided a correct classification of cases of 97.4%. Our results do not support the hypothesis of PD as secondary to anxiety disorders and confirm previous findings of a lack of specificity between DSM-III-R axis II categories and OCD and PAD.

Morbidity risk for mood disorders in the families of borderline patients.

We analyzed the familial morbidity risk for mood disorders (MR) and the presence of a family history of alcoholism in a group of 58 patients with DSM-III borderline personality disorder (PD). The MR in the families of borderline subjects was not significantly different from that found in a control group of affective patients with other cluster II PD, or without PD. The MR in the families of borderline subjects who had never developed an affective episode was not significantly different from that found in the families of borderline PD with a history of mood disorders. Borderline subjects with mood disorders had higher rates of alcoholism in their families, mainly among parents. Our results support the hypothesis that borderline PD, even in absence of the codiagnosis of a mood disorder in the subject, may be a predictor of higher familial liability to mood disorders, although it may be more informative for the familial clustering of specific subgroups than for mood disorders as a whole.

Psychiatric disorders in the families of schizotypal subjects.

In a study of the families of 21 schizotypal patients, we found an increased morbidity risk for schizophrenia compared with that in the families of 21 nonschizotypal patients and 42 controls. The Axis I diagnoses did not influence the distribution of the morbidity risk in the families of the schizotypal patients. If the schizotypal subjects also had other personality disorders, the morbidity risk for schizophrenia among their relatives was lower, although not significantly.

Personality features related to generalized anxiety disorder.

Forty-six patients with generalized anxiety disorder (GAD) without any other coexisting axis I diagnoses were compared with 50 control subjects for assessment on axis II. No specific personality disorder (PD) was found to be significantly associated with chronic anxiety, although the majority of anxious patients showed significantly more maladaptative traits than controls. Discriminant analysis selected a list of items able to provide a correct classification rate of 91% based on personality features selected in canonical function. Factor analysis indicated that personality characteristics of expectation of damage were more closely related to GAD in our sample.