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Introduction: The clinician's subjective experience can be a valuable element for diagnosis and treatment. A few factors have been recognized that affect it, such as the patient's personality, the severity of psychopathology, and diagnosis. Other factors, such as patient's and clinician's gender, have not been specifically investigated. The aim of this study is to explore the impact of gender differences on the clinician's subjective experience in a large sample of psychiatric patients., Methods: The study involved 61 psychiatrists and 960 patients attending several inpatient and outpatient psychiatric settings. The clinicians completed the Assessment of Clinician's Subjective Experience (ACSE) questionnaire after observing each patient for the first time., Results: In multivariate analysis, higher scores on the Difficulty in Attunement (p < 0.001), Engagement (p<0.05), and Impotence (p<0.01) scales were significantly associated with female clinician gender, whereas higher scores on the Tension and Disconfirmation scales were significantly associated with male clinician gender. The scores on all ACSE dimensions were also associated with the severity of psychopathology., Conclusion: The findings suggest that clinician's gender might affect a clinician's emotional response toward patients. Specific attention to this issue might be useful in clinical situations, not only in terms of promoting gender-balanced teams but also in terms of enhancing self-observation in clinicians evaluating patients for the first time., (© 2021 Dazzi et al.)

Introduction: The last decade has witnessed a resurgence of interest in the clinician's subjectivity and its role in the diagnostic assessment. Integrating the criteriological, third-person approach to patient evaluation and psychiatric diagnosis with other approaches that take into account the patient's subjective and intersubjective experience may bear particular importance in the assessment of very young patients. The ACSE (Assessment of Clinician's Subjective Experience) instrument may provide a practical way to probe the intersubjective field of the clinical examination; however, its reliability and validity in child and adolescent psychiatrists seeing very young patients is still to be determined., Methods: Thirty-three clinicians and 278 first-contact patients aged 12-17 years participated in this study. The clinicians completed the ACSE instrument and the Brief Psychiatric Rating Scale after seeing the patient, and the Profile of Mood State (POMS) just before seeing the patient and immediately after. The ACSE was completed again for 45 patients over a short (1-4 days) retest interval., Results: All ACSE scales showed high internal consistency and moderate to high temporal stability. Also, they displayed meaningful correlations with the changes in conceptually related POMS scales during the clinical examination., Discussion: The findings corroborate and extend previous work on adult patients and suggest that the ACSE provides a valid and reliable measure of the clinician's subjective experience in adolescent psychiatric practice, too. The instrument may prove to be useful to help identify patients in the early stages of psychosis, in whom subtle alterations of being with others may be the only detectable sign. Future studies are needed to determine the feasibility and usefulness of integrating the ACSE within current approaches to the evaluation of at-risk mental states., (© 2021 S. Karger AG, Basel.)

Introduction: The clinical encounter is still at the core of the psychiatric evaluation. Since the diagnostic process remains basically clinical in nature, several authors have addressed the complexity of the clinical reasoning process and highlighted the role played by intersubjective phenomena and clinician's feelings. Some recent studies have supported the view of a significant link between the clinician's subjective experience during the assessment and the diagnosis made. In a globalized world, this issue requires a careful reflection, since cultural differences may affect the intersubjective atmosphere of the encounter, which may indirectly influence the clinician's thinking., Methods: We used a previously validated instrument, named Assessment of Clinician's Subjective Experience (ACSE), to compare the clinician's subjective experience during the evaluation of Italian patients with the subjective experience of the same clinician during the assessment of foreign patients. The 2 patient groups (n = 42 each) were individually matched for known potential confounders (age, sex, categorical diagnosis, and clinical severity)., Results: We found no significant differences in mean scores on all ACSE dimensions (tension, difficulty in attune-ment, engagement, disconfirmation, and impotence), which suggests that cultural diversity did not substantially affect the clinician's subjective experience. However, the lack of information about the native country and linguistic proficiency of about a quarter of foreign patients may have limited the possibility to detect subtle or specific differences, especially with regard to the clinician's empathic attunement., Conclusions: Although further investigation is needed, our preliminary findings may have significant implications for the reflection upon the clinician's empathic experience as well as pragmatic consequences for the act of psychiatric diagnosis in the cross-cultural encounter., (© 2020 S. Karger AG, Basel.)

Background: A link between depression and insecure attachment has long been postulated. Although many studies examined the relationship between depressive symptoms and attachment, relatively few studies were performed on patients diagnosed with depression. Also, research on patients with bipolar disorder is scarce., Objective: We aimed at testing the association between attachment insecurity and unipolar and bipolar depression., Methods: We studied 21 patients with bipolar disorder, current episode depressed, and three age- and sex-matched groups, each consisting of 21 individuals: patients with major depressive disorder, recurrent episode; patients with epilepsy; non-clinical participants. The Experience in Close Relationships questionnaire was used to assess adult attachment style., Results: Patients with both bipolar and unipolar depression displayed significantly higher scores on attachment-related avoidance as compared with patients with epilepsy and non-clinical participants. Also, patients with bipolar depression scored significantly higher on attachment-related anxiety than all other groups. In both psychiatric groups, attachment dimensions were not significantly correlated with global clinical severity or severity of depression., Conclusion: Despite some study limitations, our results are consistent with some previous studies and provide support to Bowlby's seminal hypothesis that attachment insecurity may predispose to depression. Attachment theory may provide a valuable theoretical framework for future research and for guiding treatment., (© 2019 Bentham Science Publishers.)

This study examined associations between specific elements of therapeutic relationships and short-term psychodynamic psychotherapy (STPP) outcomes. Notably, we focused on therapists' subjective experiences during their first clinical interaction with patients, countertransference patterns and therapeutic alliance evaluated early in treatment, in relation to patient symptom changes at the end of STPP. Twenty clinicians completed the Comprehensive Psychopathological Rating Scale to evaluate patients' ( N =32) symptom severity at the beginning and end of STPP. They also completed the Assessment of Clinicians' Subjective Experience (ACSE) to assess their subjective experiences of their patients at the first clinical interview and the Therapist Response Questionnaire (TRQ) and Working Alliance Inventory to evaluate their countertransference reactions and therapeutic alliance at the sixth therapy session. The findings showed that the TRQ and ACSE scales correlated in a coherent way, with the exception of the TRQ helpless/inadequate pattern and ACSE impotence. Strong and more negative TRQ countertransference patterns and ACSE dimensions were significantly associated with lower quality of the therapeutic alliance. Finally, better STPP outcomes were positively associated with a good therapeutic alliance and negatively correlated with greater difficulty in attunement at the beginning of clinical assessment and therapists' stronger responses of helplessness, frustration, and disengagement during therapy. These results confirm the precious value of the clinical relationship, which represents a useful source of information for therapists when planning therapeutic interventions., Competing Interests: Conflict of interest: the authors declare no potential conflict of interest., (©Copyright A. Tanzilli et al., 2018.)

The current debate about the diagnostic significance of delusion revolves around two positions. The neurocognitive position conceives delusion as a non-specific, though polymorphic, symptom. The psychopathological position views features of delusion such as content and structure as having meaningful connections with diagnostic entities. This study aims at contributing to this debate by examining the association between delusional themes and diagnosis in a sample of 830 adult psychotic patients. All diagnoses were made by experienced psychiatrists according to DSM-IV or ICD-10 criteria, and in 348 patients were established with the SCID-I. All patients were administered the Brief Psychiatric Rating Scale (BPRS). In each patient, the presence of somatic delusions and delusions of guilt, grandiosity, and persecution was determined by examining the scores on relevant BPRS items. Delusions of guilt were almost pathognomonic for a psychotic depressive condition (psychotic major depression 40%; psychotic bipolar depression 30%; depressed schizoaffective disorder 8%; bipolar and schizoaffective mixed states 6 and 7%, respectively). Only 1% of patients with schizophrenia and no patient with delusional disorder or bipolar or schizoaffective manic state showed such delusions. The difference between unipolar and bipolar depression and the other diagnostic groups was highly significant. Delusions of grandiosity characterized mostly patients with manic symptoms (bipolar mania 20%; bipolar mixed states 19%; manic schizoaffective disorder 10%). They were observed significantly more often in bipolar mania than in schizophrenia (7%). Persecutory delusions were broadly distributed across diagnostic categories. However, they were significantly more frequent among patients with schizophrenia and delusional disorder compared with depressed and manic patients. Somatic delusions were also observed in all diagnostic groups, with no group standing out as distinct from the others in terms of an increased prevalence of somatic delusions. Our findings suggest a middle position in the debate between the neurocognitive and the psychopathological approaches. On the one hand, the widespread observation of persecutory delusions suggests the usefulness of searching for non-specific pathogenic mechanisms. On the other hand, the association between some delusional contents and psychiatric diagnosis suggests that a phenomenological analysis of the delusional experience may be a helpful tool for the clinician in the diagnostic process.

Despite the development and widespread diffusion of modern nosographic systems, the diagnosis of schizophrenia continues to raise several epistemological issues. To address these issues, a number of researchers are currently pursuing the possibility of an integration between reliable, objective approaches and the intersubjective perspective in the clinical encounter. In the present article, we discuss Rümke's popular concept of praecox feeling, as introduced in 1941 and re-elaborated over the following 20 years. Our aim was to thoroughly analyze the author's original formulation and to identify the connections between his thinking and certain psychopathological developments, epistemological issues, and research perspectives on schizophrenia. The praecox feeling is presented by Rümke as a sensitive diagnostic tool for schizophrenia that is rooted in the peculiar subjective experience of the clinician when encountering a schizophrenic patient. This experience seems to be characterized by two essential dimensions: a subjective one, which reflects the failure of a clinician's empathic effort due to a fundamental alteration of the intersubjective space, a phenomenon related to schizophrenic autism, and a gestaltic, objective one, which is grounded in the clinician's implicit typifying process as a consequence of collecting recurrent clinical observations over the course of his/her professional experience. According to Rümke, the diagnostic use of the praecox feeling should be limited to the acute phases of the schizophrenic process, as the clinician's experience of an intersubjective struggle is attenuated in interactions with older, chronic patients. The multifaceted nature of Rümke's proposal seems to have contributed to some theoretical critiques and to inconclusive results from empirical investigations, leading to a progressive devaluation of the scientific and diagnostic validity of praecox feeling. The present analysis of the original concept suggests that a renewed research interest in the role of the clinician's subjective experience with regard to the schizophrenic patient could be helpful., (© 2018 S. Karger AG, Basel.)

Classical psychopathology highly valued the interaction between clinician and patient, and recent findings have provided preliminary evidence of an association between categorical psychiatric diagnosis and the clinician's subjective experience during the first clinical assessment. To extend these findings, the present study examined the relationship between psychopathological dimensions and clinicians' subjective experiences. The study involved 45 clinicians and 783 patients in several psychiatric inpatient and outpatient units. When they saw a new patient, the clinicians completed the Assessment of Clinician's Subjective Experience questionnaire (ACSE) and the 24-item Brief Psychiatric Rating Scale (BPRS). Scores on five core psychopathological dimensions supported by meta-analytic evidence (Affect, Positive Symptoms, Negative Symptoms, Activation, Disorganization) were derived from the BPRS. Multivariate analysis revealed that each psychopathological dimension was characterized by a distinct pattern of independent associations with certain aspects of Clinician's Subjective Experience, as measured by the ACSE. This study provided preliminary evidence of significant and theoretically consistent relationships between major psychopathological dimensions and the psychiatrist's subjective experience during the first clinical evaluation. Improving the understanding of intersubjective processes may have important implications for theory, practice, research, and training., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Classical psychopathology greatly valued the interaction between clinician and patient, and assigned to the clinician's subjective experience a significant role in the diagnostic process. Psychoanalysis, too, ascribed a privileged position to the clinician's feelings and empathic participation in the assessment and deep understanding of the patient. This study aimed at testing the traditional, though still relatively unexplored empirically, tenet that particular diagnostic groups elicit distinct and diagnostically useful reactions from clinicians., Sampling and Methods: The study was performed in several psychiatric inpatient and outpatient units in Rome, Italy. The clinicians completed the Assessment of Clinician's Subjective Experience (ACSE) questionnaire and other standardized assessment instruments when they evaluated a previously unknown patient. All adult patients diagnosed with schizophrenia (n = 119), cluster B personality disorder (n = 114), manic or mixed bipolar I episode (n = 59), and unipolar depression or anxiety disorder (n = 130) were included in the study, for a total of 422 patients evaluated by 35 clinicians., Results: We found a significant and theoretically consistent relationship between the clinicians' pattern of subjective experience during the first visit and patients' clinical diagnoses. Patients with unipolar depression/anxiety showed significantly lower scores than the other groups on all ACSE scales except engagement; patients with schizophrenia displayed significantly higher scores than the other groups on difficulty in attunement, and significantly higher scores than patients with cluster B personality disorder on impotence. Compared with the other groups, the patients with cluster B personality disorder displayed significantly lower scores on engagement, and significantly higher scores on disconfirmation. In multivariate models controlling for patient's age and education, symptom severity, clinician's sex, duration of visit and setting, diagnosis remained a significant predictor of scores on all ACSE scales except for impotence., Conclusions: The main limitations of the study are its reliance on clinical diagnoses and the non-independence of assessments. Further studies based on diagnoses made by a third observer through standardized instruments are needed to provide a most stringent test of the hypothesis that different diagnoses are associated with distinct profiles of clinicians' subjective experience. This study provided intriguing, though preliminary, evidence that the clinician's subjective experience may play a useful role in the diagnostic process. Time may have come to reintroduce the concept of intersubjectivity at the core of the diagnostic process., (© 2016 S. Karger AG, Basel.)

Background: While psychiatric literature has shown renewed interest in fine psychopathological investigation, little study has been devoted to the clinician's subjective experience with the patient, which is highly valued by the phenomenological and psychodynamic traditions. We aimed at developing a valid and reliable instrument to measure such experience., Sampling and Methods: First, 104 self-report items were developed, based on daily clinical practice and references from the literature on clinician's subjective experience. Of these, 46 were retained after pilot testing and exclusion of items with poor psychometric properties. Thirteen psychiatrists and 527 first-contact patients participated in the validation study. Psychiatrists completed the 'Assessment of Clinician's Subjective Experience' (ACSE) instrument and the Brief Psychiatric Rating Scale (BPRS) after the visit and the Profile of Mood State (POMS) before and after it. The ACSE was completed again for 60 patients with stable BPRS scores over a short retest interval. Principal component analysis with orthogonal rotation was performed. The internal consistency and test-retest stability of ACSE factorially derived scales were calculated. Convergent validity was tested by examining the correlations between ACSE scores and change in POMS scores during the visit., Results: Five factors (interpreted as tension, difficulty of attunement, engagement, disconfirmation, impotence) accounting for 57% of total variance were extracted. All ACSE scales showed high internal consistency and stability, and correlated with conceptually related POMS scales., Conclusions: The pattern of subjective experience identified by the ACSE is consistent with classical psychopathological descriptions and previous related studies. Despite limitations such as the relatively small number of psychiatrists studied and the exclusively self-report nature of the instrument, this study supports the validity and reliability of the ACSE and suggests that it may be a valuable tool for training, research and possibly diagnostic purposes., (Copyright © 2013 S. Karger AG, Basel.)

Aim: In this work it is discussed whether and how Kretchmer's psychopathological reflections about sensitive delusion of reference can offer a relevant interpretative key for clinicians who face cases of acute persecutory psychosis. It is argued the utility and topicality of those psychopathological concepts that aren't commonly investigated, especially in the evaluation and management of onset paranoid psychosis in an emergency ward., Methods: We provide clinical vignettes of two young patients, admitted in the emergency psychiatry ward, who represent a concrete example of the dynamic-affective comprehension of delusional elaboration, through its embodiment in the individual biographical development., Results: An interpretative key, based on this specific conceptual frame, seems to provide an integrated intervention tool that aims both to a causal comprehension and demolition of the delusional solution in onset psychotic cases., Conclusions: The rediscovery of classical psychopathological concepts appears to be a necessary process, especially in the clinical management of onset psychotic disorders. Further and more accurate researches are, in any case, needed.

Aims: A robust psychopathological and psychotherapeutic tradition underscores the importance of the clinician's feelings in the assessment and therapeutic process. Our aim is to develop an instrument to evaluate psychiatrist' experience induced by each patient. This paper describes the development and preliminary validation of this instrument., Methods: Based on the relevant literature and our clinical experience, we developed a self-completed questionnaire consisting of 65 items covering various aspects of the clinician' emotional resonance. Ten psychiatrists completed our questionnaire immediately after first clinical interviews involving a total of 125 patients with various psychiatric diagnoses. Also, they completed the Profile of Mood State (POMS) immediately before and after these interviews. The factor structure and convergent validity of the questionnaire was examined., Results: We identified a six-factor structure. The factors showed acceptable internal consistency and were interpretable as different aspects of the clinician-patient interaction as experienced by the clinician. There was a consistent pattern of correlations between factor scores and changes in POMS scale scores during the clinical interviews., Discussion: The study results, though preliminary, suggest that clinicians' subjective reactions during their interactions with patients can be validly and reliably measured. Quantitative measurement of clinician's subjectivity is potentially useful in both clinical and research settings.

Laughter is a very common behaviour in everyday life, nevertheless scientific literature is lacking in studies which examine closely its nature. The study aims are: to summarise the present knowledge about laughter and its relation with depression and to make hypotheses on its possible therapeutic function. In the first part of the review the main data existing about encephalic structures involved in laughter genesis, which show participation of cortical and subcortical regions, are reported and the effects of laughter on the organism physiologic equilibrium, particularly on the neuroendocrine and immune systems, are described. In the second part, scientific evidence about the influence of depression on the ability to laugh are referred, which suggests that reduction of laughter frequency is a symptom of the disease and that its increase may be used as a marker of clinical improvement. Finally, the main assumptions supporting the hypothesis of the therapeutic action of laughter on depression are examined: first of all, it has been demonstrated that laughter is able to improve mood directly and to moderate negative consequences of stressful events on psychological well-being; in addition, it is possible that the stimulation of particular cerebral regions, involved in depression pathogenesis, and the normalisation of the hypothalamic pituitary adrenocortical system dysfunctions, both mediated by laughter, can counteract efficiently depressive symptoms; finally, the favourable effects of laughter on social relationships and physical health may have a role in influencing the ability of depressed patients to face the disease.

Intermediate filaments are frequently used in studies of developmental biology as markers of cell differentiation. To assess whether they can be useful to identify differentiating pancreatic endocrine cells, we examined the pattern of expression of nestin, cytokeratin 20, and vimentin on acetone-fixed cryosections of rat adult and developing pancreas. We also studied vimentin expression in mouse embryonic pancreas at E19. Cytokeratin 20 was found in all pancreatic epithelial cell lineages during the entire development of the rat gland and in the adult animals. Under our experimental conditions, therefore, cytokeratin 20 is not an exclusive marker of rat duct cells. Nestin was detected exclusively in stromal cells either in the adult or developing rat pancreas. Vimentin was observed within cells located in the primitive ducts of rat pancreas starting from E12.5. Their number rapidly increased, reaching its highest level in newborn animals. Vimentin was also spotted in alpha cells starting from E12.5 but disappeared soon after birth, likely identifying immature or recently differentiated alpha cells. In addition, vimentin was observed in duct and alpha cells of mouse developing pancreas showing that its expression in such cells is not an event restricted to the rat. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.

Nestin is considered a marker of neurogenic and myogenic precursor cells. Its arrangement is regulated by cyclin-dependent kinase 5 (CDK5), which is expressed in murine podocytes. We investigated nestin expression in human adult and fetal kidney as well as CDK5 presence in adult human podocytes. Confocal microscopy demonstrated that adult glomeruli display nestin immunoreactivity in vimentin-expressing cells with the podocyte morphology and not in cells bearing the endothelial marker CD31. Glomerular nestin-positive cells were CDK5 immunoreactive as well. Western blotting of the intermediate filament-enriched cytoskeletal fraction and coimmunoprecipitation of nestin with anti-CDK5 antibodies confirmed these results. Nestin was also detected in developing glomeruli within immature podocytes and a few other cells. Confocal microscopy of experiments conducted with antibodies against nestin and endothelial markers demonstrated that endothelial cells belonging to capillaries invading the lower cleft of S-shaped bodies and the immature glomeruli were nestin immunoreactive. Similar experiments carried out with antibodies raised against nestin and alpha-smooth muscle actin showed that the first mesangial cells that populate the developing glomeruli expressed nestin. In conclusion, nestin is expressed in the human kidney from the first steps of glomerulogenesis within podocytes, mesangial, and endothelial cells. This expression, restricted to podocytes in mature glomeruli, appears associated with CDK5.

Human adrenocortical cells have been shown to express cytokeratins and vimentin. Nestin is an intermediate filament protein that is mainly expressed in the developing nervous system and that has been recently reported in rat adrenal gland as well. Using immunohistochemical and biochemical approaches, the present study demonstrates that nestin is constantly expressed in situ in the cortex of normal human adrenal glands. Nestin expressing cells were prevalently located in the zona reticularis but some positive cells could be spotted in the zona fasciculata as well. Moreover, patches of nestin-positive cells have been constantly detected on sections of cortical adenomas. In contrast, adrenal carcinomas displayed a variable number of nestin-immunoreactive cells that in some cases were virtually absent. Samples of renal clear cell carcinoma metastasis in the adrenals were also examined which did not show nestin-immunoreactivity. We propose that a positive nestin-immunoreaction could be useful in differential diagnosis of clear cell tumors in adrenal glands.

The activation of the molecular cascade leading to Ca++ -induced differentiation in cultured epithelial cells might be provided by the establishment of intercellular junctions between cells. In the present paper, we tested the hypothesis that Ca++ concentration would determine morphological and biochemical changes in intercellular junctions of cultured human gingival cells. Triplicate samples of monolayer cultures of human oral gingival cells were grown with two different Ca++ concentrations (0.3 and 1.8 mM), and examined by transmission (TEM) and scanning (SEM) electron microscopy at different time periods. To determine the role of the E-cadherin/beta-catenin complex in intercellular junction formation, oral epithelial cell cultures were grown in 0.3 mM Ca++ in presence of a blocking antibody anti human E-cadherin, stained with antibodies anti human beta-catenin, and examined by confocal laser scanning microscopy (CLSM). By TEM and SEM, cells grown at physiologic Ca++ concentrations (i.e., 1.8 mM) showed a subjective increase of the size of microvilli and of the number of intercellular junctions, which was more evident after 3 days in culture. Desmosome-like junctions were observed in cells grown in 1.8 mM Ca++, not in cells grown in 0.3 mM. By CLSM, development of intercellular adhesion was marked by membranous localization of E-cadherin and beta-catenin within the first hours in both culture types. When cell-cell adhesion was prevented, cells showed round shape and no membranous localization of beta-catenin. Restoring cell adhesion brought about polygonal cell shape and membranous localization of beta-catenin. We can conclude that increased Ca++ concentration may determine biochemical and morphological changes at membranous level in human oral epithelial cells. These changes may facilitate the development of intercellular junctions.

In the past three decades, many studies have analyzed ultrastructural and molecular markers of differentiation in squamous stratified epithelial tissues. In these tissues, epithelial cells migrating from the basal layer to the upper layers undergo drastic changes, which involve membrane-associated proteins, DNA synthesis, phenotypic aspects, lipid composition, and cytoskeletal components. Cytoskeletal components include a large and heterogeneous group, including intermediate filaments, components of the cornified envelope, and of the stratum corneum. When grown in mono- and multilayer cell cultures, epithelial cells isolated from the oral mucosa may reproduce many of the biochemical and morphological aspects of epithelial tissue in vivo. In the present paper, we examine phenotypic changes, development of suprabasal layer, and Involucrin expression occurring in differentiating oral epithelial cells, based on literature review and original data.

Despite many studies on the topic, plasma cells found in human periapical chronic inflammatory lesions (granulomas) continue to present unresolved issues. In this study, we tried to assess quantitatively and qualitatively the nature of plasma cells of 4 human periapical granulomas. Samples were analyzed for relative amounts of IgG-, IgM-, IgA-, and IgE-positive plasma cells by immunohistochemistry, and for morphological changes by transmission electron microscopy (TEM). By immunohistochemistry, many plasma cells stained positively with anti-IgG and anti-IgM antibodies; fewer cells reacted with anti-IgE and anti-IgA. Russell Bodies, controversial aspects of plasma cell maturation, showed positive reactivity of the superficial layer only to antibodies against IgG and IgM. By TEM analysis, phenotypes of normal and dysfunctional plasma cells (Mott cells) were evident. Russell Bodies appeared as intra- or extracellular round vesicles, with an homogeneous internal core, and an external membrane, resembling rough endoplasmatic reticulum (RER). We can conclude that mucosal immune response is not the predominant type in the periapical lesions examined. Positive immunoreaction for IgG and IgM of Russell Bodies may be due to the residual RER membrane, whereas components of yet unidentified nature may occupy the internal core.

M-cells are believed to play a pivotal role in initiation of the immune response. These cells, located in the epithelia that overlie mucosal lymphoid follicles, are responsible for the active uptake of particulate antigens and for their translocation to the underlying lymphoid tissue. The identification of reliable markers for M-cells is therefore extremely important for the study of the initial steps that lead to the immune response. For this purpose, we studied cytokeratin 20 (CK20) expression in the epithelium of rabbit palatine tonsils by immunofluorescence, confocal microscopy, and Western blotting. CK20+ cells were observed in all rabbit palatine tonsils examined. By Western blotting, one CK20-immunoreactive band was identified at 46 kD on samples of proteins from the intermediate filament-enriched cytoskeletal fraction of tonsil epithelium. Double labeling of CK20+ cells with cell-specific markers confirmed that such cells were actually M-cells. Moreover, CK20+ M-cells displayed a mature phenotype (they formed pockets harboring lymphoid cells) and were functionally competent because they could take up particulate antigens from the pharyngeal lumen. We conclude that CK20 is an M-cell marker for rabbit palatine tonsils. Moreover, we can hypothesize the use of M-cells as a possible site for antigen delivery of particle-based vaccines., (Copyright The Histochemical Society, Inc.)

Biocompatibility of metals for dental use was tested using a three-dimensional model consisting of oral keratinocytes cultured on de-epidermised sub-mucosa. The toxicity of orthodontic metallic wire and soldering material was assessed through parameters such as the morphology and growth rate of the keratinocytes, as well as by classical histology, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The sharp composition of metallic wires and soldering materials was assessed by Auger Electron Spectroscopy (AES). The results of our experiment showed that the new model revealed inhibition of keratinocyte growth and stratification near soldering material, whereas mucosal cells were able to grow and layer out on dental wire. It is concluded that this experimental model, which simulates the oral environment, is useful for studying the effects of materials for dental use for its sensibility and reproducibility. Moreover it can provide morpho-functional information which cannot be achieved by traditional methods.

Quantitative and qualitative control of oral bacterial flora is a major issue in oral pathology and in the prophylaxis against cavities. Recent findings suggest that it is possible to induce local immune responses delivering antigens on palatine tonsils. M cells play an important role in the start of the immune response. These cells are located in the epithelia overlaying mucosal lymphoid follicles and are responsible for the uptake of particulate antigens. The identification of reliable markers for M cell is therefore extremely important. Since it has been reported that tonsillar immunization leads to the secretion of high levels of specific salivary antibody, we undertook a study to identify a marker for tonsillar M cells in order to plan strategies of oral immunization against oral pathogens. We studied cytokeratin 20 expression in rabbit tonsils by immunofluorescence and confocal microscopy. Cytokeratin 20 immunoreactive cells were observed in all samples examined. These cells were identified as M cells as they co-expressed vimentin, a well-known marker of rabbit M cells, and they actively uptook particulate material. It is therefore possible to hypothesize the use of tonsil M cells as a possible site for antigen delivery of particle-based vaccines against oral pathogens.

The sleep of adults and childrens is often disturbed from obstructive respiratory desorders evidenced from snoring. Scientific literature agrees in considering that as a dangerous pathology called roncopathy. Statistics show that about 50% of adult population over 50 yrs snores (exspecially males) and some of that has a dangerous period of prolonged and frequent obstructive sleep apneas. Results of polisomnographic tests show that 4% of total population suffer of apneas extremely severe and dangerous for life. Roncopathy is often associated to a lot of troubles: pulmonary, gastroenterologic, endocrinologic, of behaviour and especially cardiovascular and neurologic and this could explain the motif of high nocturnal percentage of myocardial infarction, ictus and sudden death. In this work we evidence the anatomical and functional etiology of snoring and sleep apneas and expose the surgical and medical therapeutic options up to day available.

Renin and angiotensinogen have been previously found in the rat pancreas, and angiotensin receptors have been located in the apical domain of duct cells. To evaluate the possibility that angiotensin II could be generated within the duct system, we decided to determine whether angiotensinogen is present in rat pancreatic juice and the angiotensinogen-immunoreactive pancreatic cell types that could be responsible for its production. Angiotensinogen was detected in significant amounts by Western blotting in pancreatic juice collected from several individual rats. Different isoforms between plasma and pancreatic juice angiotensinogens were demonstrated by isoelectric focusing. Immunocytochemical experiments revealed angiotensinogen-immunoreactive cells at the periphery of the islets of Langerhans, and confocal microscopy demonstrated that most angiotensinogen-immunoreactive cells were glucagon-secreting cells. Secretion of angiotensinogen did not follow the regulated secretory pathway since it was absent from the glucagon-containing granules. This was confirmed by electron microscopy immunocytochemistry. Duct and acinar cells did not express angiotensinogen at an immunocytochemical detectable level. The present findings indicated an exocrine secretion of angiotensinogen by glucagon-secreting cells and suggest that one of the final targets of the local pancreatic renin-angiotensin system may be the duct epithelium.

In order to evaluate the influence of specific factors on mercury (P-Hg) levels and antioxidant power (P-FRAP) in human plasma, 26 healthy donors were examined by a dentist, their plasma analyzed for Hg by atomic absorption spectrometry and for total antioxidant activity by FRAP method. Hg plasma concentration was found to be correlated with the number of amalgam fillings, suggesting that Hg released from fillings is a source of Hg in non-occupational exposed subjects. P-FRAP correlated negatively with P-Hg suggesting a pro-oxidant role of the Hg released from amalgam fillings. Though age by itself was not significantly correlated with P-FRAP, when considered together with P-Hg in multivariate analysis, it was found to be a major related cofactor. Multivariate analysis showed no influence of fish consumption or cigarette smoking on P-FRAP., (Copyright 2002 Elsevier Science B.V.)

In this study, we tested the hypothesis that a significant difference exists in the integrated optical density (IOD) of membrane and cytoplasmic antigens in monolayer keratinocytes cultures. Oral normal and two malignant (SCC15 and SCC25) keratinocyte cultures were stained with antibodies specific for E-cadherin, beta-catenin, beta-1 integrin, cytokeratin (CK) 14, CK19, CK10/11 and involucrin. The IOD recorded (n = 12) was analyzed for significant differences using a two-way analysis of variance (significance level set at alpha = 0.05); antibodies and cell cultures were grouped according to Tukey's Group Comparison post-test. The majority of normal cell cultures exhibited E-cadherin, beta-catenin, involucrin and beta-1 integrin IOD values significantly higher than the two SCC cell lines. No definite staining pattern distinguished normal and malignant cells in relation to cytokeratins 14, 19 and 10/11. Our observations suggest that IOD measures constitute a good predictor of antigen steady state levels in monolayer cell cultures. According to these observations, SCC cell lines and normal cells appear to differ in the expression of E-cadherin, beta-catenin, beta-1 integrin and involucrin, although some variability within normal cells can also be observed.

The Authors of this work propose to give an evaluation about actual relationship between posture and occlusion, trying of to give some indications about posture's meaning, organizing the relation about five points: Posture's definition Posture's history Posture's models of study Occlusal support en posture Diagnostic procedures.

During embryonic and foetal development, the masseter is formed from two successive generations of muscle fibers in a manner which is very similar to that which has been previously described for other skeletal muscles. This phenotype is characterised by the persistence of ontogenic myosin isoforms (embryonic and foetal myosin heavy chains, embryonic light chain) and by the presence of two distinct populations of fibers: small diameter fibers which coexpress the embryonic, foetal and fast isoforms of the myosin heavy chains but never express the slow isoform; large diameter fibers which express the slow myosin heavy chain either exclusively or in variable associations with the other isoforms. These characteristics of the human masseter muscle probably correspond not only to its embryological origin and its special innervation, but also to the functional constraints to which it is submitted after birth.

The constituents of the intermediate filament network of adrenal gland cells have not been deeply investigated in vivo. Adrenocortical cells have been reported to express cytokeratins and vimentin, but the intermediate filament components of the adrenomedullary cells are still unknown. Nestin is an intermediate filament protein that is mainly expressed in the developing nervous and muscle systems. It has been reported to be unable to form filaments by itself and it co-assembles with vimentin. Using immunocytochemical and biochemical approaches, the present study demonstrates that nestin is expressed in situ either in the cortex or in the medulla of adult rat adrenal glands. Nestin-negative cells prevalently form the zona glomerulosa whereas the zona fasciculata and the zona reticularis are mainly nestin-immunoreactive. Nestin-positive cells always express vimentin-like immunoreactivity but several cells apparently expressing only vimentin are detectable too. Nestin is also expressed by adrenomedullary cells that also display a faint vimentin-like immunoreactivity. We hypothesise that the inconstant detection of nestin in adrenocortical cells depends on their different functional moments. Moreover, even though our data do not allow to confirm vimentin in adrenomedullary cells, in situ detection of nestin in the adrenal medulla indirectly supports in vivo expression of vimentin in chromaffin cells.

The purpose of this study was to observe the morphological and histological changes on the root canal walls after Nd:YAG laser application. Twenty vital, recently extracted single-rooted human teeth were used for this study. Root canals were cleaned and shaped by a conventional step-back technique--by means of k files up to a 20 k-file type at working length--and subsequently shaped by Ni-Ti root-canal rotary instrumentation up to 30/06 and irrigated with 2.5% hypochlorite solution. Ten teeth (control group) were left unlased, while the other ten teeth were irradiated with Nd:YAG laser by means of a 320 microns fibre inserted in the root canal at 1 mm from the apex with a power of 1.5 Watt and a frequency of 15 pps for five seconds in retraction with rotating movements. The control specimen showed debris and smear layer on the root canal surface obscuring the dentin tubules. The root canal walls irradiated with Nd:YAG laser showed a clear glazed surface, some open dentinal tubules and some surface craters with cracks. Such results confirm that smear layer and debris are removable with Nd:YAG laser, however clearing all root canal walls is still difficult and, if the energy level and duration of application are inadequate, a certain degree of thermal damage and morphological changes in dentin structure are observable.

Meckel's cartilage plays an important role in the topographical organisation and in the differentiation of the facial structure during the embryonal and even much later during the foetal period. Our observations on serial sections carried out in two human foetuses aged 12 and 16 weeks indicate that the two dorsal (tympanic) and ventral (mandibular) branches of Meckel's cartilage are perfectly defined at 16 weeks. In the dorsal branch, the primordia of the incus and of head of the malleus are still composed on non-ossified cartilage. In the ventral branch, it is also possible to describe at 16 weeks three posterior, medial and anterior parts which are composed of cartilage. The initiating role played by the ventral part of Meckel's cartilage on the ossification of the mandible leads during the embryonal period to the formation of the mandibular primary growth center, which is therefore clearly defined in our first stage at 12 weeks. The partial fibrous evolution and the regression of the major part of the ventral branch of Meckel's cartilage only start after 16 weeks of intrauterine life.

The lymphatic network of the pancreas has been little investigated and recent studies have provided contrasting data. This research is aimed to supply the morphologic basis to outline the involvement of the lymphatic system in pancreatic pathology. Guinea pigs, rats, and mice were anesthetized with ether and sacrificed with the same anesthetic. Pieces of pancreas were processed for transmission electron microscopy. Semithin sections were observed by light microscopy and, after positive identification by transmission electron microscopy, lymphatics were followed with long series of consecutive sections to define their distribution. Lymphatics were detected in the pancreas of all the animals both in the inter and the intralobular sites. Closer relations with the exocrine parenchyma (ducts and acini) were observed in guinea pig pancreas. Remarkably, interesting relationships between lymphatics and endocrine tissue were observed in all the animals. Overall, however, the lymphatic network of rat pancreas was less develop and preferentially associated with blood vessels. The distribution of the pancreatic lymphatic network appears consistent with an active role in pancreatic pathology., (Copyright 2001 Wiley-Liss, Inc.)

In order to evaluate the influence of mercury (Hg) levels on antioxidant power in human plasma, 26 healthy people were evaluated by a dentist and their plasma analyzed for Hg content by atomic absorption and total antioxidant activity (TAA) by FRAP method. Hg plasma concentration correlated with number of amalgam restorations, suggesting that Hg released from fillings is a source of Hg in non-occupational exposed people. Fish consumption, in fact, showed no influence on Hg plasma levels, perhaps because Italian subjects examined in the present group used low quantity of fish at week or kinds of fish with light contamination. TAA negatively correlated with Hg plasma revealing a pro-oxidant role of Hg released from amalgam fillings.

At the moment, in most countries, there are laws in force which impose to the manufacturers well regulated testing in order to investigate and guarantee an acceptable biocompatibility of medical devices before their commercialization. Many international laboratories are committed to the definition of investigation methodologies and to the evaluation of biocompatibility in order to obtain research standards, capable to provide reproducible and comparable objective quantitative data. In every country, technical committees were put together for a standardization of methodological procedures, followed by European and international technical boards which proposed and codified methodologies and investigation approaches. UNI-EN-ISO laws contain all the results and constitute a reference point for any consideration on or evaluation of the biocompatibility of a medical device. Based on these laws, we evaluated the biocompatibility and determined the physical-mechanical characteristics of the new Venezia (Cabon S.p.A.) endodontic ZOE sealer. The Subcutaneous Implant Technique according Safavi et al. (in vivo test, ISO 10993: 1-6 Biological evaluation of medical and dental materials and devices) and Autian test of Emolysis on Rabbit Erythrocytes (in vitro test) allowed us to evaluate a good biocompatibility of the new product. Furthermore, its Setting and Working time, its radiopacity, Solubility and its Flow value completely satisfy the requirements of international standards (ISO/DIS 6876 Dental root Canal Sealing Materials). We can finally deduce that Venezia fulfil the ideal functional properties of an endodontic cements.

Dental amalgam (AMG) is the most diffused dental filling material. Since it is constituted for at least 40-45% of Hg, many questions have raised about its safe use. Hg particles from dental amalgam dissolve in saliva and, being ingested, they reach the blood stream through the intestinal mucosa. It has been demonstrated that amalgam fillings continuously release Hg vapour and that there is detectable Hg in expired and inspired air of amalgam owners. It is not yet fully accepted that AMG fillings represent the principal source of Hg for man and the aim of this study was to evaluate if the mercury level in saliva: 1) was higher within people bearing dental amalgam restorations than in people with no restorations; 2) was different between males or females; 3) increased in relation to the surface of amalgam restorations. The results showed a correlation between number of fillings and salivary Hg, between amalgam surface and salivary Hg. The Authors could finally assert that AMG fillings represented the principal source of salivary Hg in the subjects studied.

Dental amalgam fillings are known to release significant levels of mercury (Hg) in saliva which could represent a continuous source of oxidative damage to tissues. The present investigation was aimed at verifying this hypothesis by determining a possible correlation between salivary Hg levels and salivary total antioxidant activity (TAA), used as an index of oxidative stress. Samples of saliva from 34 healthy donors were analyzed for Hg content, through vapor atomic absorption spectrometry, and for TAA, by determining the ferric reducing ability ('FRAP' method). A significant correlation between Hg and the number of amalgam restorations or total amalgam surface was evident in both the male and female subjects. A significant negative correlation between TAA and Hg levels or number of amalgam restorations or amalgam surface was evident in females, indicating that small increases in salivary Hg were sufficient to produce a decrease in salivary TAA. On the other hand, no significant correlation was found in the males. The present study provides, for the first time, evidence of a pro-oxidant role of the amalgam Hg chronically released in saliva.

This study intended to verify, through microbiological techniques and TEM investigations, the killing of bacterial spores after treatment in steam autoclave, and to propose strictly morphological considerations about the target of this sterilisation process. Autoclave is the most common device for sterilising instruments in order to prevent cross infections in dental offices. The autoclave efficiency has been improved in the last years and part of this improvement is related to both a better and more correct use of the autoclave system and to the technological innovations introduced in the last generation of devices. However, associations as ADA or CDC suggest to regularly verify the process of 'autoclaving' through biological indicators (BI). The most commonly used BI are made of spores strips or suspensions of Bacillus Subtilis (pb 168) and Bacillus Stearothermophilus (ATCC 10149). They visually prove, changing colours on enzymatic base, the death of micro-organism and if the physical parameters, necessary for sterilisation, have been achieved. These two strains of endospore-forming bacteria were processed and prepared following two different techniques: Karnovsky fixed and epon embedded--phosphotungstic acid fixed for direct observation. The kind and the extent of analysed modifications are extremely various: from deep lacerations, which changed the spore structure, to little clefts which let the cytoplasm go out.