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Background: Periodontal care, which was completely suspended during the peak of the SARS-Cov-2 pandemic and was delayed and recurrently discontinued during the succeeding waves, must be safely provided in the COVID-19 era. Objectives: The study aimed to provide recommendations on periodontal practice, optimizing safety, ergonomics, and economic resources, and the management of periodontal patients, ensuring continuity, timing, and effectiveness of periodontal care in the COVID-19 era. Methods: Reported protocols for the dental practice in the context of the COVID-19 and current evidence on periodontitis treatment and prevention were reviewed. Results: Evidence-based recommendations on contamination control and ergonomic improvements for periodontal practice and the management of periodontal patients challenging COVID-19 and airborne infectious diseases have been provided. Conclusion: Due to the economic, ergonomic, and ethical concerns raised by limited periodontal care due to the SARS-Cov-2 pandemic, and awareness of other emerging airborne transmitted infections, the periodontal practice should integrate measures minimizing airborne cross-infections and optimizing time-space and economic resources. The management of periodontal patients in the COVID-19 era should comprise less complex treatments and more comprehensive and definitive approaches, reducing emergencies, session length and number, and, concurrently, extending the recall and maintenance intervals. Moreover, it should implement prevention strategies through teledentistry tools and apps, improving periodontal awareness and self-care, and also through the self-reporting of periodontitis and periodontal risk assessment tools, performing both “population-based” and “high-risk” surveillance of periodontitis. Finally, it should enhance inter-professional collaboration, through telehealth networks, especially targeting subjects at high-risk of both periodontitis and systemic disorders, each of the two variously linked to COVID-19 onset and worsening.

Complex periprosthetic cases are considered as challenges by clinicians. Clinical and radiographic parameters should be considered separately to make the right choice between an endodontically or periodontally compromised treated tooth and implant. Therefore, in order to decide whether the tooth is safe or not, data that have to be collected are specific parameters of both the patient and the clinician. In addition, the presence of periodontal, prosthetic, and orthodontic diseases requires patients to be set in multidisciplinary approach. The aim of this case report is to describe how the multidisciplinary approach could be the best way to manage difficult cases of implant-prosthetic rehabilitation. How to rehabilitate with fixed prosthesis on natural teeth and dental implants a smoker patient who presents with active periodontitis, multiple edentulous areas, dental malocclusion, and severe aesthetic problems was also described.

Purpose To assess the accuracy of a combined Scheimpflug camera–Placido disk device (Sirius, CSO, Italy) in evaluating corneal power changes after myopic photorefractive keratectomy (PRK). Methods Two hundred and thirty-seven eyes of 237 patients that underwent myopic PRK with a refractive error, measured as spherical equivalent, ranging from −10.75 D to −0.5 D (mean −4.63 ± 2.21 D), were enrolled in this study. Corneal power evaluation using Sirius were performed before, 1, 3 and 6 months after myopic PRK. Mean simulated keratometry (SimK) and mean pupil power (MPP) were measured. Correlations between changes in corneal power, measured with SimK and MPP, and variations in subjective refraction, calculated at corneal plane, were evaluated using Pearson test at every follow up; differences between preoperative and postoperative data were evaluated with the Student paired t-test. Results A good correlation has been detected between the variations in subjective refraction measured at corneal plane 1, 3 and 6 months after myopic PRK and both SimK (R2 = 0.8463; R2 = 0.8643; R2 = 0.7102, respectively) and MPP (R2 = 0.6622; R2 = 0.5561; R2 = 0.5522, respectively) but corneal power changes are statistically undervalued for both parameters (p Conclusions Even if our data should be confirmed in further studies, SimK and MPP provided by this new device do not seem to accurately reflect the changes in corneal power after myopic PRK.

Purpose: To analyze and compare intraocular pressure (IOP) values measured in healthy subjects (HS), keratoconus (KC) patients and patients that underwent myopic photorefractive keratectomy (REF), using Goldmann applanation tonometry (GAT), dynamic contour tonometry (DCT), ocular response analyzer (ORA) and Corvis ST (CST). Methods: The study included 76 eyes of 76HS, 15 eyes of 15 KC patients and 18 eyes of 18 subjects that underwent REF. Each participant underwent a complete ophthalmic evaluation, IOP measurement with GAT, DCT, ORA and CST. Results: HS showed a mean GAT value of 15.62 ± 2.33 mm Hg, a mean DCT value of 17.44 ± 2.51 mm Hg, a mean ORA value of 15.99 ± 3.58 mm Hg and a mean CST value of 17.24 ± 3.44 mm Hg. KC showed a mean GAT value of 15.07 ± 1.83 mm Hg, a mean DCT value of 17.01 ± 1.96 mm Hg, a mean ORA value of 13.58 ± 2.99 mm Hg and a mean CST value of 14.37 ± 1.89 mm Hg. REF showed a mean GAT value of 14.06 ± 1.51 mm Hg, a mean DCT value of 15.12 ± 2.34 mm Hg, a mean ORA value of 16.85 ± 2.4 mm Hg and a mean CST value of 15.57 ± 1.77 mm Hg. Conclusion: Our data suggest that ORA and GAT could be used interchangeably in HS; GAT, ORA and CST could be used interchangeably in KC patients and that GAT provides lower IOP values compared to the other devices in eyes previously submitted to myopic PRK. Purpose: To analyze and compare intraocular pressure (IOP) values measured in healthy subjects (HS), keratoconus (KC) patients and patients that underwent myopic photorefractive keratectomy (REF), using Goldmann applanation tonometry (GAT), dynamic contour tonometry (DCT), ocular response analyzer (ORA) and Corvis ST (CST). Methods: The study included 76 eyes of 76HS, 15 eyes of 15 KC patients and 18 eyes of 18 subjects that underwent REF. Each participant underwent a complete ophthalmic evaluation, IOP measurement with GAT, DCT, ORA and CST. Results: HS showed a mean GAT value of 15.62 ± 2.33 mm Hg, a mean DCT value of 17.44 ± 2.51 mm Hg, a mean ORA value of 15.99 ± 3.58 mm Hg and a mean CST value of 17.24 ± 3.44 mm Hg. KC showed a mean GAT value of 15.07 ± 1.83 mm Hg, a mean DCT value of 17.01 ± 1.96 mm Hg, a mean ORA value of 13.58 ± 2.99 mm Hg and a mean CST value of 14.37 ± 1.89 mm Hg. REF showed a mean GAT value of 14.06 ± 1.51 mm Hg, a mean DCT value of 15.12 ± 2.34 mm Hg, a mean ORA value of 16.85 ± 2.4 mm Hg and a mean CST value of 15.57 ± 1.77 mm Hg. Conclusion: Our data suggest that ORA and GAT could be used interchangeably in HS; GAT, ORA and CST could be used interchangeably in KC patients and that GAT provides lower IOP values compared to the other devices in eyes previously submitted to myopic PRK.

Summary Background Serum and IgG isolated from patients with the autoimmune blistering disease pemphigus vulgaris (PV) trigger complex intracellular pathways in keratinocytes, including alterations of the cell cycle and metabolism, which ultimately lead to cell–cell detachment (acantholysis). We have shown previously that one of the earliest pathogenic events in PV is the activation of protein kinases, including the PKR-like endoplasmic reticulum (ER) kinase PERK. Objectives In the present study we investigated in more detail the role of PERK in the pathogenesis of PV. Methods PERK levels were assessed by Western blotting and in-cell enzyme-linked immunosorbent assay, and PERK expression was silenced by siRNA technology. The effects of PV sera/IgG on keratinocyte cultures were investigated by flow cytometry, MTT and adhesion assays. Results We show that PERK is activated in keratinocytes exposed to PV serum, as demonstrated by an increase in phosphorylated PERK levels and phosphorylation of eIF2α. Decreased expression of PERK by siRNA reduced the effects of PV serum on the cell cycle and keratinocyte viability, two key events in PV pathophysiology. As impairment of metabolic activity in PV is partially due to non-IgG serum factors, we then investigated the activation of PERK in keratinocytes incubated with whole PV serum, purified PV IgG and IgG-depleted PV serum. The data demonstrated that PV sera depleted of IgG, but not PV IgG, triggered PERK phosphorylation and this correlated with a marked reduction of metabolic activity in keratinocytes exposed to IgG-free serum. Knockdown of PERK by siRNA abrogated the changes in the cell cycle and apoptosis induced by IgG-depleted PV serum. Finally, the reduction of metabolic activity observed in keratinocytes exposed to IgG-depleted PV serum was almost absent in PERK-deficient cells. Conclusions Taken together, the results demonstrate that activation of PERK participates in the reduction of metabolic activity and cell viability seen in PV and that this phenomenon depends on non-IgG factors. PERK activation may represent a novel signalling mechanism linking ER stress and acantholysis in PV.

Objectives: Pemphigus vulgaris (PV) is an autoimmune blistering disease affecting primarily oral mucosa and skin. Among the drugs used for the therapy of pemphigus, both methylprednisolone (MP) and pyridostigmine bromide (PBr) can prevent acantholysis in vitro. However, their putative therapeutic properties in regenerating PV-like lesions and promoting the healing process still remain to be demonstrated. To address this issue, here we have developed a model for studying the process of epithelial cleft regeneration in PV by artificially wounding keratinocyte monolayers. Materials and methods: The experimental model was established by scratching confluent monolayers to simulate the epithelial cleft; then, wound regeneration in the presence of submaximal concentrations of PV sera was studied by time-lapse microscopy, with or without the addition of MP and PBr in the culture medium. Results: Pemphigus vulgaris serum inhibited epithelial cleft repair of wounded monolayers. Indeed, in the presence of 10% (v/v) PV serum, keratinocytes reached only 2% confluence within 72 h vs an almost complete healing of controls. When administered together with PV sera, MP significantly (P

We have previously demonstrated that serum autoantibodies of patients with pemphigus vulgaris (PV) may affect desmoglein 3 (Dsg3)-mediated adhesion by decreasing its half-life and inducing Dsg3 cleavage. Here we sought to gain more insights into the role of Dsg3-targetting IgG in acantholysis. To do so, alterations of keratinocyte morphology and cell–cell adhesion strength were investigated in the presence of PV serum, PV IgG, and IgG purified from PV patients’ sera against linear epitopes of Dsg3 (anti-Dsg3-L IgG). Changes in Dsg3 protein levels were assessed by Western blotting. Results showed that both PV serum and PV IgG were able to induce acantholysis and decrease the total amount of Dsg3 in cell lysates. Polyclonal anti-Dsg3-L IgG displayed Dsg3-depleting activity solely when used at 1 μg/ml, i.e. under non-physiologic conditions. Furthermore, cell–cell detachment induced by PV IgG and anti-Dsg3-L IgG seemed to precede the loss of Dsg3 from keratinocytes, suggesting that depletion/degradation of Dsg3 represents a late event in acantholysis. Collectively, the data presented here demonstrate that PV IgG recognizing non-conformational epitopes of Dsg3 are pathogenic when administered on doses largely exceeding those found in PV sera.

Aphthous ulcers are the most common oral mucosal lesions in the general population. Several precipitating factors for aphthous ulcers are suggested to operate on subjects with genetic predisposition. Sometimes aphthous ulcers can be the sign of systemic diseases. Therefore, it is essential to establish a correct diagnosis to determine suitable therapy. There are several diseases potentially responsible for oral ulcers. Sometimes appearance of periodic oral ulcers coincides with periodic fever and other symptoms leading to the diagnosis of a rare childhood disease: PFAPA (periodic fever, aphthous stomatitis, pharyngitis and adenopathy) syndrome. PFAPA or Marshall's syndrome is characterized by abrupt onset of periodic episodes of high fever accompanied by aphthous stomatitis, pharyngitis and cervical adenitis, often associated with headache and / or abdominal or joint pain. Owing to the periodic onset of oral symptoms, often an oral physician or pediatric dentist may be the first healthcare worker to evaluate a child with clinical signs compatible with PFAPA syndrome. Children diagnosed with this condition require systematic oral follow-up to monitor for signs of ulceration.

A wide range of diseases may present with radiographic features of osteolysis. Periapical inflammation, cysts and benign tumours, bone malignancies, all of these conditions may show bone resorption on radiograph. Features of the surrounding bone, margins of the lesion, and biological behaviour including tendency to infiltration and root resorption, may represent important criteria for distinguishing benign tumours from their malign counterpart, although the radiographic aspect of the lesion is not always predictive. Therefore a critical differential diagnosis has to be reached to choose the best management. Here, we report a case of giant cell tumour (GCT) whose radiological features by computed tomography (CT) suggested the presence of bone malignancy, whereas the evaluation of a routine OPT scan comforted us about the benign nature of the lesion. A brief review of the literature on such a benign but locally aggressive neoplasm is also provided.

Primary oral melanoma (POM) is an uncommon malignant tumor that originates from the proliferation of melanocytes. Such tumors can be present at any location in the oral cavity; however, it affects more frequently the hard palate and the maxillary alveolar mucosa. POM is usually asymptomatic in the early stages and it presents normally as a pigmented patch or as a mass with a rapid growth rate. In the advanced stages, it can show ulceration, swelling, bleeding, rapid enlargement and loosening of teeth. Melanoma of the mouth is rare, most commonly occurring in the upper jaw of patients more than 65 years. Because of a frequent delay in diagnosis, the tumors are often diagnosed when they are deeper than the average cutaneous melanoma. The prognosis is extremely poor, especially in advanced stages. Therefore, pigmented lesions of undetermined origin should be routinely subjected to a biopsy examination. In this study, we aimed to present a review on primary malignancy.

Intercellular adhesion among keratinocytes is guaranteed by desmosomes. Disruption of desmosomal integrity leads to cell-cell detachment or acantholysis, as it classically occurs in pemphigus vulgaris (PV), an autoimmune blistering disease of skin and mucous membranes. While purified PV IgG seems to trigger intracellular signaling that crucially involves p38 MAPK, keratinocyte acantholysis induced by whole PV serum may recruit a number of additional signals. In this study, the Pro-Q® Diamond Phosphoprotein Assay was used to investigate the overall changes in protein phosphorylation levels in an in vitro model of PV. We showed that keratinocytes exposed to whole PV sera underwent at least three early and transient phosphorylation events. Two bands with apparent molecular masses of ∼35 and ∼45 kDa were found to be phosphorylated within 1 min after incubation with PV sera. A third band of about 80 kDa reached the peak of phosphorylation level after 3 hours. Morphologic evidence of cell shrinkage and acantholysis were late events and did not correlate temporally with kinase activation, suggesting that cytoskeleton reorganization is a downstream phenomenon. Interestingly, pharmacological abrogation of PV-specific protein phosphorylation was able to inhibit the cell-cell detachment, rounding up, and redistribution of Dsg3 in keratinocytes. Thus, at least three phosphorylation events are pathogenically involved in pemphigus acantholysis.

Aphthous ulcers are the most common oral mucosal lesions in the general population. These often are recurrent and periodic lesions that cause clinically significant morbidity. Many suggestions have been proposed but the etiology of recurrent aphthous stomatitis (RAS) is unknown. Several precipitating factors for aphthous ulcers appear to operate in subjects with genetic predisposition. An autoimmune or hypersensitivity mechanism is widely considered possible. Sometimes aphthous ulcers can be the sign of systemic diseases, so it is essential to establish a correct diagnosis to determine suitable therapy. Before initiating medications for aphthous lesions, clinicians should determine whether well-recognized causes are contributing to the disease and these factors should be corrected. Various treatment modalities are used, but no therapy is definitive. Topical medications, such as antimicrobial mouth-washes and topical corticosteroids (dexamethasone, triamcinolone, fluocinonide, or clobetasol), can achieve the primary goal to reduce pain and to improve healing time but do not improve recurrence or remission rates. Systemic medications can be tried if topical therapy is ineffective.

Pemphigus vulgaris (PV) blistering occurs as a result of the disruption of intercellular contacts among keratinocytes, or acantholysis. The hallmark of PV acantholysis in vitro is considered to be the retraction of keratin intermediate filaments (KIF) onto the nucleus, which parallels with loss of cell-cell adhesion and rounding up of keratinocytes. However, the fine morphological changes of keratinocytes as well as the fate of cell adhesion structures cannot be appreciated on immunofluorescence by the simple cytokeratin staining. In this paper, we show that acantholytic dysmorphisms are sharply investigated by using PV IgG as a primary antibody on metabolically quiescent living cells. Indeed, PV IgG recognise a wide spectrum of molecules and enabled us to monitor the main changes occurring in acantholytic keratinocytes, including cell shrinkage with the appearance of prickle-like processes, detachment of keratinocytes from one another and collapse of cytoskeleton-bound proteins along nuclear periphery. This method has wider applications as it could be useful for staining cell periphery of keratinocytes and changes in cell shape. Furthermore, images displayed clear and sharp contours because living cell microscopy allows to avoid antigen distortion due to cell manipulation, which usually precedes the immunolabelling. © 2007 Springer-Verlag.

Serum antibodies against desmoglein 1 (Dsg1) are known to induce the clinical and histological manifestations of pemphigus foliaceus (PF), autoimmune bullous disease targeting skin. The basic pathophysiological phenomenon of PF blistering is the disruption of epithelial integrity in the granular layer of the epidermis due to separation of keratinocytes from one another, or acantholysis. In this report we investigate the changes in subcellular distribution of Dsg1 in response to serum of patients with PF by using an in vitro model of PF. Immunofluorescence analysis on HaCaT cells indicates that non-clustered Dsg1 is markedly internalized after exposure to serum. However, binding of PF IgG to Dsg1-rich adhesion complexes (desmosomes) does not cause disruption of such structures nor depletion of clustered Dsg1, as revealed by colocalization of PF IgG and Dsg1 in a punctate staining on cell membrane 24 hours after treatment. Furthermore, morphological studies demonstrate that the dramatic alterations induced by PF sera are not the result of apoptotic programs. Taken together, our data strongly suggest that anti-Dsg1 antibodies from PF serum could cause the internalization of non-clustered Dsg1 and perturb the formation of new desmosomes but not directly disrupt Dsg1-containing junctions when stable contacts are already formed.

Pemphigus vulgaris (PV) is considered to be an autoimmune disease affecting skin and mucous membranes. Traditionally, PV autoantibodies are thought to recognize antigens located in the intercellular substance (ICS) of keratinocytes; antigens represented mainly by the desmosomal cadherin desmoglein 3 (Dsg3). Accordingly, titres of anti-ICS and anti-Dsg3 immunoglobulin G (IgG) are considered to be major laboratory criteria when making a diagnosis of PV. In this paper, we demonstrated for the first time that PV IgG bind antigen(s) expressed on the surface of peripheral blood mononuclear cells (PBMC), as revealed by immunofluorescence studies. This novel autoantigen is immunoprecipitated by PV IgG as a 130 000 molecular weight protein. However, Western blot analysis of the immunocomplexes failed to show reactivity with anti-Dsg3 monoclonal and polyclonal antibodies. Taken together, our data provide strong evidence that PV autoimmunity targets a 130 000 antigen other than Dsg3 on PBMC. This shifting from epidermis to blood cells may open new perspectives for a better understanding of pemphigus autoimmunity and more rational approaches to its treatment.

Pemphigus vulgaris (PV) is a disease of epidermal adhesion. Its pathogenesis is currently traced back to the action of autoantibodies against antigens located within the intercellular substance of keratinocytes, such as desmogleins and acetylcholine receptors. In the present paper, we sought to elucidate the non-IgG-mediated effects of PV sera on keratinocytes. Results showed that PV sera depleted of IgG were able to induce well-defined changes on keratinocyte morphology and metabolic activity. Indeed, PV IgG-free sera determined marked alterations on cell shape, accompanied by partial loss of keratinocyte–keratinocyte interactions within 48 h after treatment. Furthermore, PV IgG-depleted sera caused a sharp reduction of cell viability along with a less sustained weakening of intercellular adhesion strength. In light of the above findings, loss of cell–cell adhesion in PV occurs as a result of the cooperating action of both IgG and non-IgG-mediated mechanisms. These data have remarkable consequences on experimental models of PV and might open new “biological” approaches to its therapy. Thus, researchers are well advised that PV pathophysiology cannot be faithfully reproduced by leaving non-IgG serum factors out of consideration. J. Cell. Physiol. 212:563–567, 2007. © 2007 Wiley-Liss, Inc.

PURPOSE: To evaluate the correlation between corneal biomechanical and morphological data in healthy eyes. METHODS: A complete clinical eye examination of naïve eyes was followed by tomographic (Pentacam, Oculus, Wetzlar, Germany) and biomechanical (Corvis ST, Oculus, Wetzlar, Germany) evaluation. Linear regression between central corneal thickness (CCT), corneal volume (CV) and anterior corneal curvature measured with Sim'K (SK), versus corneal deformation parameters measured with Corvis ST have been run using SPSS software version 18.0. RESULTS: Seventy-six eyes of 76 healthy subjects (44 women and 32 men) with a mean age of 36.84±10.74 years and a mean refractive error of -0.55±1.68D (measured as spherical equivalent) were evaluated. Corneal deformation parameters were weakly correlated with corneal morphological parameters and with spherical equivalent. Although the correlations between deformation amplitude versus SK and between SK versus Velocity of Applanation 2, were higher than the others (R2=0.28 and 0.26 respectively), none of them was statistically significant (p>0.01). CONCLUSIONS: According with these findings, Corvis ST seems to be able to provide an analysis of corneal deformation independent from corneal morphological characteristics. If these data will be confirmed in further studies, this device could be useful in the management and screening of eyes with corneal diseases. Copyright © 2014 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

The aim of this paper is to describe a case of implant-prosthetic rehabilitation in a patient with periodontitis, focusing on the different timing of implant placement. After initial periodontal treatment, teeth with advanced mobility degree and severe bone resorption were extracted. At different healing time oral implants were placed in a prosthetic-guided position. After osseointegration period the implants were loaded and the results at one year of follow-up are presented. The aim of this paper is to describe a case of implant-prosthetic rehabilitation in a patient with periodontitis, focusing on the different timing of implant placement. After initial periodontal treatment, teeth with advanced mobility degree and severe bone resorptionwere extracted.At different healing timeoral implantswere placed in a prosthetic-guided position.After osseointegration period the implants were loaded and the results at one year of follow-up are presented.

Background: Pemphigus vulgaris is a life-threatening autoimmune blistering disease targeting skin and mucous membranes, characterized by disruption of keratinocytes’ adhesion termed acantholysis. Today multiple classes of targets are considered to play a role in the genesis of the acantholysis; of these, the classical pemphigus antigens, desmosomal cadherins (desmoglein 1 and 3) are the best characterized and considered as the most important. Additional antigens include the novel epithelial acetylcholine receptors (a9 and pemphaxin). Thus, acantholysis in pemphigus seems to result from a cooperative action of antibodies to different keratinocyte self-antigens, but the mechanisms by which epithelial cleft occurs are not yet clearly understood. In fact, the binding of the autoantibodies to these targets generates a plethora of biological effects due, on one hand, to their direct interference with adhesive function and, on the other, to more complex events involving intracellular pathways that modify proteases activity or calcium metabolism, leading to loss of cell–cell adhesion.

This experiment tested the involvement of the ventromedial hypothalamus (VMH) in the sympathetic and hyperthermic reactions induced by an intracerebroventricular (i.c.v.) injection of orexin A (1.5 nmol). In the first part of the experiment, the firing rate and cytochrome oxidase activity of the VMH neurons, and the colonic temperature were monitored in 12 urethane-anaesthetized rats before an i.c.v. injection of orexin and over a period of 2 h after the injection. Orexin induced an increase in the firing rate, colonic temperature and cytochrome oxidase activity. A group of 12 rats was used as a control: saline, but not orexin, was injected. No modifications in the firing rate, cytochrome oxidase reactivity and colonic temperature were noted. In the second part of the experiment, 12 rats were anaesthetized and lesioned bilaterally in the VMH with an injection of ibotenic acid. Sham lesions were carried out in 12 control rats. After 48 h, all animals were anaesthetized with ethyl-urethane. The firing rates of the sympathetic nerves to interscapular brown adipose tissue (IBAT), along with IBAT and colonic temperatures and heart rate were monitored before and over a period of 2 h after an i.c.v. injection of orexin or saline in the lesioned and sham-lesioned rats. Orexin increased the sympathetic firing rate, IBAT and colonic temperatures and heart rate in the sham-lesioned rats. These increases were reduced by lesion of VMH. Saline did not induce any modification. These findings indicate that the VMH is involved in the control of the orexin-induced hyperthermia.

A novel explanation of pemphigus vulgaris (PV) pathogenesis suggests that serum autoantibodies may affect desmoglein 3 (Dsg3)-mediated adhesion by triggering depletion of Dsg3 from desmosomes. Furthermore, abrogation of Dsg3 from the cell seems to depend on anti-Dsg3 pemphigus IgG. In this study we sought to gain more insights into the role of PV IgG recognizing non-conformational epitopes of Dsg3 (anti-Dsg3-L IgG) by semi-quantitative living cell immunofluorescence (LCIF) microscopy, in-cell ELISA and morphometric analysis of acantholysis. Our data demonstrate that PV serum and PV IgG can induce acantholysis and reduce the total amount of Dsg3 in cultured keratinocytes, whereas anti-Dsg3-L IgG fail to do so when administered at concentrations comparable to those present in pathogenic PV sera. However, the Dsg3-depleting activity of such polyclonal anti-Dsg3 IgG was acquired when used at 1μg/ml. Interestingly, both PV sera and IgG, including anti-Dsg3-L IgG, caused early depletion of surface Dsg3 while slightly affecting the total cell content of Dsg3 until late acantholysis. This raises a possibility that depletion of Dsg3 from cell membrane and reduction of the total cellular levels of Dsg3 represent distinct phenomena in PV acantholysis. Taken together, our data demonstrate that anti-Dsg3 PV IgG against linear epitopes of Dsg3 can induce acantholytic changes of keratinocytes in a dose- and time-dependent manner. Specifically, both morphological and biochemical changes suggestive of acantholysis are seen only at high IgG concentrations. We conclude that anti-Dsg3L IgG play a minor role in experimental PV under physiologic conditions.

Objective: We aimed to compare the efficacy of saliva substitutes and citric acid long-term therapy for oral dryness relief and unstimulated salivary flow in patients reporting drug-induced xerostomia. Study design: Fifty-four patients reporting drug-induced xerostomia were randomly subdivided into 3 groups and respectively administered artificial saliva, 3% citric acid, or distilled water in mouthwash 4 times a day for 30 days. Patients underwent measurement of unstimulated whole saliva before and after they finished therapy and were asked to note in a daily diary any symptomatologic changes 15 minutes and 1 hour after each daily intake of test solution. Results: Fifteen minutes after solution intake, 12 patients (67%) belonging to the artificial saliva group, 9 (50%) from the citric acid group, and 2 (11%) from the water group reported significant symptomatologic improvement. One hour after solution intake, 7 patients (39%) from the artificial saliva group, 10 (56%) from the citric acid group, and 0 from the water group noted significant symptomatologic improvement. None of the drugs tested affected unstimulated whole saliva flow. Conclusions: Both artificial saliva and citric acid provided immediate relief from oral dryness. Citric acid also provided a longer-lasting feeling of oral moistness at 1 hour after use owing to its protracted activity on salivary gland function. © 2011 Mosby, Inc. All rights reserved. Objective: We aimed to compare the efficacy of saliva substitutes and citric acid long-term therapy for oral dryness relief and unstimulated salivary flow in patients reporting drug-induced xerostomia. Study design: Fifty-four patients reporting drug-induced xerostomia were randomly subdivided into 3 groups and respectively administered artificial saliva, 3% citric acid, or distilled water in mouthwash 4 times a day for 30 days. Patients underwent measurement of unstimulated whole saliva before and after they finished therapy and were asked to note in a daily diary any symptomatologic changes 15 minutes and 1 hour after each daily intake of test solution. Results: Fifteen minutes after solution intake, 12 patients (67%) belonging to the artificial saliva group, 9 (50%) from the citric acid group, and 2 (11%) from the water group reported significant symptomatologic improvement. One hour after solution intake, 7 patients (39%) from the artificial saliva group, 10 (56%) from the citric acid group, and 0 from the water group noted significant symptomatologic improvement. None of the drugs tested affected unstimulated whole saliva flow. Conclusions: Both artificial saliva and citric acid provided immediate relief from oral dryness. Citric acid also provided a longer-lasting feeling of oral moistness at 1 hour after use owing to its protracted activity on salivary gland function. © 2011 Mosby, Inc. All rights reserved.

In confluent keratinocyte monolayers, desmosomal adhesion gradually becomes calcium-independent and this is associated with an increase in the strength of intercellular adhesion (hyper-adhesion). In this study, we investigated the functional and molecular significance of hyper-adhesion in a system challenged by autoimmune sera from patients with Pemphigus Vulgaris (PV), a disease primarily targeting desmosomal adhesion. The results show that keratinocytes with calcium-independent desmosomes are resistant to disruption of intercellular contacts (acantholysis) in experimental PV. Furthermore, both the desmosomal cadherins desmoglein (Dsg) 1 and Dsg3 and the adherens junction protein E-cadherin were decreased in confluent keratinocytes at Day 1, but not in hyper-adhesive cells (Day 6) after incubation with PV serum. Pharmacological induction of the hyper-adhesive state with the PKC inhibitor Go6976 reduced both the acantholysis rate and the processing of cell adhesion molecules induced by PV serum. When the establishment of the hyper-adhesive state was prevented by cell adhesion recognition (CAR) peptides that perturbed desmosomal interactions, Go6976 could still partially attenuate PV acantholysis. Taken together, these data demonstrate that keratinocyte hyper-adhesion decreases the morphological, functional and biochemical dys-cohesive effects of PV serum via mechanisms that involve, at least in part, the function of PKC. This suggests that reinforcing keratinocyte adhesion may be a promising way to inhibit the effects of this most debilitating disorder.

Hirsutism is the presence of terminal hair in women in a male-like pattern, which represents the clinical expression of an underlying excess of androgen (hyperandrogenism). Numerous conditions and serious diseases can result in high levels of circulating androgens, although the most common cause of hirsutism is polycystic ovary syndrome (PCOS). The following classic clinical features characterize PCOS: irregular menstrual periods, infertility, excess hair growth, and obesity. The diagnosis of this disease is related to the discovery of clinical signs that are confirmed by hematochemical examinations. We present a clinical case that is relatively unusual owing to the appearance of black hairs that are similar to nasal hair in the oral mucosa, which is an atypical location. This unusual case was considered to be oral hirsutism, and its presence constituted the essential element in the diagnosis of PCOS. The hypothesis proposed for this unusual condition could be correlated to the high rate of circulating testosterone that may have influenced and led to the development and appearance of the hair follicles in the oral mucosa. Hirsutism is the presence of terminal hair in women in a male-like pattern, which represents the clinical expression of an underlying excess of androgen (hyperandrogenism). Numerous conditions and serious diseases can result in high levels of circulating androgens, although the most common cause of hirsutism is polycystic ovary syndrome (PCOS). The following classic clinical features characterize PCOS: irregular menstrual periods, infertility, excess hair growth, and obesity. The diagnosis of this disease is related to the discovery of clinical signs that are confirmed by hematochemical examinations. We present a clinical case that is relatively unusual owing to the appearance of black hairs that are similar to nasal hair in the oral mucosa, which is an atypical location. This unusual case was considered to be oral hirsutism, and its presence constituted the essential element in the diagnosis of PCOS. The hypothesis proposed for this unusual condition could be correlated to the high rate of circulating testosterone that may have influenced and led to the development and appearance of the hair follicles in the oral mucosa. © 2009 Mosby, Inc. All rights reserved.

We have previously demonstrated that serum of patients with pemphigus vulgaris induces reduction of desmoglein 3 (Dsg3) half-life in keratinocytes (FEBS Lett 2006: 580: 3276). This phenomenon seems to occur as a consequence of the progressive depletion of Dsg3 from desmosomes. Here we reported that reduction of full-length Dsg3 may be due to its progressive cleavage, leading to the formation of two fragmentation products with apparent molecular masses of about 60 kDa (fragment 1) and 70 kDa (fragment 2), as revealed by Western blotting. Unexpectedly, analysis of fragmentation pattern suggested cleavage to occur intracellularly. Consistently, fragment 1 was shed and localized within the cytosol, as shown by living cell immunofluorescence microscopy. Total amounts of full-length plakoglobin and Dsg1 were apparently unchanged. Taken together, our findings provide evidence that proteolytic processing of Dsg3 can lead to depletion of Dsg3 from the cell.

We know that the Yin Yang 1 protein (YY1) overexpression is positively and strongly correlated with the degree of malignancy of bone tumors. Therefore, we questioned whether we could influence cell invasiveness by deleting YY1 in human osteosarcoma cells (SaOs-2), as tested in Matrigel-coated filters and metastasis implantation of such osteosarcoma cells in vivo, by serial analysis with nuclear magnetic resonance. Moreover, we focused our work on the chemokine receptor CXCR4 and its inhibition by T22 antibody, as well as on systemic (direct in vivo assay) and computer-assisted imaging of angiogenesis-related metastasis. Results showed that cell invasiveness and metastasis implantation by wild-type SaOs-2 cells, as evaluated by histology and immunohistochemistry, are associated with up-regulation of CXCR4 expression, which in turn was significantly reduced by T22. In addition, deletion of YY1 (siRNAYY1-SaOs-2) induced a significant decrease of cell invasion and metastasis growth. This phenomenon was associated with decreased vascular endothelial growth factor (VEGF)/angiogenesis and a complex rearrangement of the gene expression profile as evaluated by microarray analysis. In conclusion, YY1 and VEGF/CXCR4 seem to intervene in the pathogenesis of the malignant phenotype of osteosarcoma by acting on cell invasiveness and metastasis growth. [Cancer Res 2008;68(6):1797–808]

Background Adverse drug reactions are noxious and unintended responses to a medicinal product. Many drugs have the potential to induce adverse reactions in the mouth. The extent of such reactions is unknown; however, because a lot of them are asymptomatic, many are believed to go unnoticed. Adverse oral drug reactions are responsible for oral lesions and manifestations that can mime local or systemic disease. Their pathogenesis, especially of the mucosal reactions, is largely unknown and appears to involve complex interactions between the drug in question, other medications, the patient’s underlying disease, genetics and lifestyle factors. Aim In this study, we have listed the principal signs and symptoms of oral and perioral adverse drug reactions and the responsible drugs. Diagnosis for adverse drug reaction is not easy given also the limited utility of laboratory tests. The association between a drug and an adverse drug reaction is mostly based on the disappearance of the reactions following discontinuance of the offending drug. Sometimes, it is useful to perform rechallenge tests reintroducing the drug to establish cause and effect. Conclusions Knowledge of adverse drug-induced oral effects helps health professionals to better diagnose oral disease, administer drugs and improve patient compliance during drug therapy and may foster a more rational use of drugs.

The pathogenesis of pemphigus vulgaris (PV) is still poorly understood. Autoantibodies present in PV patients can promote detrimental effects by triggering altered transduction of signals, which results in a final acantholysis. To investigate mechanisms involved in PV, cultured keratinocytes were treated with PV serum. PV sera were able to promote the cell cycle progression, inducing the accumulation of cyclin-dependent kinase 2 (Cdk2). Microarray analysis on keratinocytes detected that PV serum induced important changes in genes coding for one and the same proteins with known biological functions involved in PV disease (560 differentially expressed genes were identified). Then, we used two different approaches to investigate the role of Cdk2. First, small interfering RNA depletion of Cdk2 prevented cell-cell detachment induced by PV sera. Second, pharmacological inhibition of Cdk2 activity through roscovitine prevented blister formation and acantholysis in the mouse model of the disease. In vivo PV serum was found to alter multiple different pathways by microarray analysis (1463 differentially expressed genes were identified). Major changes in gene expression induced by roscovitine were studied through comparison of effects of PV serum alone and in association with roscovitine. The most significantly enriched pathways were cell communication, gap junction, focal adhesion, adherens junction, and tight junction. Our data indicate that major Cdk2-dependent multiple gene regulatory events are present in PV. This alteration may influence the evolution of PV and its therapy. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc.

Pemphigus is a chronic, autoimmune disease involving the skin and Malpighian mucous membranes. Pemphigus leads to progressive blistering and subsequent erosions. This article describes the etiology and treatment of pemphigus.

Despite advances in imaging, understanding the underlying pathways, and clinical translation of animal models of disease there remains an urgent need for therapies that reduce brain damage after stroke and promote functional recovery in patients. Blocking oxidant radicals, reducing matrix metalloproteinase-induced neuronal damage, and use of stem cell therapy have been proposed and tested individually in prior studies. Here we provide a comprehensive integrative management approach to reducing damage and promoting recovery by combining biological therapies targeting these areas. In a rat model of transient cerebral ischemia (middle cerebral artery occlusion) gene delivery vectors were used to overexpress tissue inhibitor of matrix metalloproteinase 1 and 2 (TIMP1 and TIMP2) 3 days before ischemia. After occlusion, autologous bone marrow cells alone or in combination with agents to improve NO bioavailability were administered intraarterially. When infarct size, BrdU incorporation, and motor function recovery were determined in the treatment groups the largest beneficial effect was seen in rats receiving the triple combined therapy, surpassing effects of single or double therapies. Our study highlights the utility of combined drug, gene, and cell therapy in the treatment of stroke. © 2007 by The National Academy of Sciences of the USA.

Pemphigus vulgaris (PV) acantholysis represents a complex phenomenon wherein a number of factors cooperates. PV serum is known to modulate important cellular events, including kinase activity, transcriptional regulation, and proteinase expression. Indeed, transduction of signals to the cell triggered by PV serum may induce proteinase up-regulation potentially responsible for disruption of epidermal adhesion and, ultimately, blister formation. Here, we sought to investigate this hypothesis by using both in vivo and in vitro models of PV. Microarray analysis on mouse skin tissues suggested that the equilibrium between extracellular proteinases and their inhibitors moved towards enhanced proteolytic activity in PV neonatal mouse model, at least on the transcriptional level. Conversely, genes codifying cell adhesion proteins were dramatically down-regulated. The effects of PV serum on the protein level were then studied in vitro both in keratinocyte monolayers and skin organ cultures focusing on matrix metalloproteinase (MMP) 9 expression and activity. By means of Western blotting, zymography, and living cell immunofluorescence studies, we showed that MMP-9 was early overexpressed in keratinocytes exposed to PV serum, and subsequently secreted in the culture medium. However, we failed to demonstrate extracellular activation of MMP-9, since it was found in its 92 kDa inactive form in serum-free culture supernatants. Taken together, our data demonstrated that proteinase expression, particularly of MMP-9, is modulated by PV serum and associated with PV acantholysis.

The current knowledge on Pemphigus vulgaris (PV) pathophysiology suggests that blister formation relies on both PV IgG and non-IgG serum factors activity. PV autoimmunity seems to develop against both desmoglein 1/3 and acetylcholine receptors leading to transduction of signals to the cell mediated by phosphorilation events. Serum factors other than IgG also participate to PV acantholysis through apoptotic or cytokine-mediated mechanisms. Apart from the role played by each actor within the acantholysis, however, the current scenario arises important methodological issues. For example, the use of PV IgG or monoclonal anti-Dsg3 antibodies to experimentally reproduce the disease appears inadequate, as it does not take into account the role of non-IgG factors. On the basis of the above observations and those from our laboratories, here we propose that using whole sera from PV patients with active disease represents the most faithful manner to mimic the disease.

Within the last decade, a number of theories on the pathogenesis of pemphigus vulgaris (PV) have followed one another. Of these, plesminogen activation and desmoglein compensation hypotheses have been substantiated by a conspicuous body of evidence. A significant change of this scenario occurred with the discovery that autoimmunity in PV can target acetylcholine receptors and that PV serum elicits a pletora of intracellular signals. Since then, a myriad of explanations accounting for PV acantholysis have appeared in the literature. However, as revolutionary as they can be, the majority of organic theories seemed to be highly speculative. We have recently obtained evidence for a proteolytic cleavage of desmoglein 3 in an in vitro model of PV; furthermore, our previous findings suggested the possible involvement of proteases such as matrix metalloproteinase (MMP) 9 in PV acantholysis both in vitro and in vivo. Hence, in formulating the "specific proteolysis theory" we have kept the rationale and the well-established evidence of both plasminogen activation and desmoglein compensation hypotheses. However, the specific proteolysis theory proposed by us is not just a return to the past. On the basis of the current knowledge on MMP substrate specificity we propose that Dsg1 and Dsg3, along with other important cadherins which are likely to be proteolytically targeted in PV, could be cleaved by either ADAM or typical MMPs, respectively. Whether this view was confirmed by further investigations, these enzymes could be specifically targeted by selective drugs which would permit more rational approaches to the treatment of pemphigus. © 2007 Elsevier Ltd. All rights reserved.

The complexity of pemphigus acantholysis together with the weak expression of desmoglein 1 (Dsg1) in cultured keratinocytes have made the study on the pathogenic action of anti-Dsg1 antibodies quite difficult. The pathophysiology of the acantholytic phenomenon could depend on the reduction of Dsg1 adhesion function occurring after its massive internalization or decrease of its synthesis. Here, we have investigated this hypothesis by using sera of patients having antibodies against Dsg1 or monoclonal anti-Dsg1 antibodies to simulate pemphigus autoimmunity in Dsg1-rich keratinocytes. Similar to pemphigus foliaceus (PF) and vulgaris (PV) sera, monoclonal anti-Dsg1 antibodies induced transient internalization of Dsg1 and reduced the adhesion strength among keratinocytes. However, binding of IgG to Dsg1 did not determine its early depletion from the adhesion complexes but reduced the amount of Dsg1 found in the Triton X-100 soluble pool of proteins. Taken together, our results represent the first demonstration that anti-Dsg1 antibodies induce similar alterations on the subcellular distribution of Dsg1 irrespective of the disease where they come from. Furthermore, the present study provides insight into the mechanisms underlying epithelial blistering observed in the skin type of pemphigus.

Moderate physical exercise (PE) combined with metabolic treatment (MT) (antioxidants and l -arginine) are well known to reduce atherosclerotic lesion formation in hypercholesterolemic mice. However, the long-term beneficial effects on unstable atheroma remain poorly understood. We started early PE training in large groups of 6-week-old hypercholesterolemic mice (by graduated swimming) alone or in combination with nutritional supplementation (1.0% vitamin E added to the chow and 0.05% vitamin C and 6% l -arginine added to the drinking water). Inactive controls did not receive PE. The spontaneous development of atherosclerotic plaque rupture (associated with advanced atherosclerosis) and survival rates were evaluated. Moderate PE elicited an increase in plasma levels of nitric oxide. Early combined treatment with PE and MT in the hypercholesterolemic mice significantly reduced lesions (also detected noninvasively at 10 months) and spontaneous atherosclerotic plaque rupture and prolonged survival more effectively than each intervention alone. Thus, early concerted actions of MT and PE improve the natural history of atherosclerotic lesions and reduce the plaque instability in hypercholesterolemic mice.

Defects of cell–cell adhesion underlie disruption of epithelial integrity observed in patients with pemphigus vulgaris (PV), an autoimmune disease characterized by severe mucosal erosions and skin blisters. Pathogenic PV autoantibodies found in patients’ sera target desmoglein 3 (Dsg3), a major component of the desmosome, but how does this phenomenon affect Dsg-dependent adhesion and lead to acantholysis still remains controversial. Here, we show that PV serum determines a reduction of Dsg3 half-life in HaCaT keratinocytes, although the total amount of Dsg3 remains unchanged. Immunofluorescence studies suggest that PV IgG exert their effect prevalently by binding non-desmosomal Dsg3 without causing its massive internalization. Furthermore, PV IgG targeting desmosome-assembled Dsg3 do not induce depletion of Dsg3 from the adhesion sites. Conversely, incorporation of PV IgG-Dsg3 complexes into new forming desmosomes appears perturbed. With our study, the basic biochemical changes of Dsg3 in an in vitro model of PV have been defined.

Background: Gorham’s disease is a rare disorder characterized by spontaneous and progressive osteolysis of one or more skeletal bones. The radiographic findings associated with Gorham’s disease are particularly dramatic, as in some cases a complete resorption of the involved bone can occur, leading to the definition of phantom bone, vanishing bone, or disappearing bone disease. Material and methods: A 24-year-old female patient with a previous diagnosis of periodontal disease and progressive mandibular alveolar bone loss was referred to our Oral Medicine section. The initial radiographic picture showed infrabony defects and horizontal bone loss. Results: After further extensive local and systemic evaluation, including histopathological, laboratory and imagine techniques investigations, the patient was diagnosed to be affected by Gorham’s diease. Meanwhile the progression of the osteolytic process had caused the loosening of all the left mandibular teeth and a pathologic fracture. Appropriate medical therapy was successful in stabilizating the resorptive process, with no evidence of further progressive disease. Conclusions: When Gorham’s disease involves the mandible, the role of the periodontologist is extremely important in diagnosing promptly the disorder and preventing the functional and aesthetic consequences of advanced and extensive bone loss. Gorham’s disease should be included among the pathologic entities mimicking periodontal disease on radiograph, such as inflammatory disease (e.g. osteomyelitis), endocrine disease (e.g. hyperparathyroidism), intra-osseous malignancies or metastases, lymphoma, histiocytosis X, mainly eosinophilic granuloma, infective process (e.g. tuberculosis and actinomycosis), odontogenic tumours.

Patients with pemphigus vulgaris (PV) may develop autoimmunity against an array of non-desmoglein antigens, including keratinocyte acetylcholine receptors. The latter, along with yet unknown autoantigens, could participate to PV pathogenesis. Here, we report a case of PV developing autoimmunity against peripheral blood mononuclear cell (PBMC) antigens. The patient was poorly responsive to steroidal and immunosuppressive drugs. Although the role for this PBMC antigen has not been established, it could affect the behaviour of immune cells that regulate autoantibody production in PV. Deregulation of PBMC activity may therefore influence patient’s response to pharmacological therapy.

Summary Patients with pemphigus vulgaris (PV) who have both antidesmoglein (Dsg)1 and anti-Dsg3 antibodies usually develop flaccid blisters on skin and mucous membranes. We report a case of PV with crusting skin lesions resembling pemphigus erythematosus, the localized variant of pemphigus foliaceus (PF). Notably, the patient had high titres of anti-Dsg1 IgG, as assessed by ELISA. We then established an in vitro model of pemphigus, and found that patient's serum was able to induce suprabasilar acantholysis in mouse skin culture. However, epidermal splitting also occurred within the granular layer, suggesting that the pathogenic potential of such a high-titre anti-Dsg1 serum was intermediate between PV and PF. Thus, the levels of anti-Dsg1 antibodies could play a role in determining the clinical phenotype of pemphigus.

Pemphigus vulgaris (PV) is considered as an autoimmune disease against a tissue-restricted antigen, desmoglein 3, a 130 kDa glycoprotein expressed by keratinocytes of skin and mucous membranes. Therefore, a breakdown of peripheral tolerance is generally invoked to explain this horror autotoxicus. The availability of a self-antigen and the strength of antigenic stimulation represent critical points in the regulation of immune system homeostasis. Our study shows for the first time that the immunodominant fraction of the PV self-antigen is present in sera of healthy individuals and patients as a circulating 30 kDa fragment (sDsg3). These findings provide a good explanation for the N-terminal specificity of antibody production and peptide recognition in PV patients by B and T cell, respectively. Moreover, the presence of the sDsg3 in human sera could allow to reconsider pemphigus as a disease against a circulating antigen; once produced, PV-autoantibodies also recognize the 130 kDa epidermal antigen desmoglein 3 on keratinocyte surface (kDsg3), thus triggering the acantholysis and the clinical manifestations of pemphigus Pemphigus vulgaris (PV) is considered as an autoimmune disease against a tissue-restricted antigen, desmoglein 3, a 130 kDa glycoprotein expressed by keratinocytes of skin and mucous membranes. Therefore, a breakdown of peripheral tolerance is generally invoked to explain this horror autotoxicus. The availability of a self-antigen and the strength of antigenic stimulation represent critical points in the regulation of immune system homeostasis. Our study shows for the first time that the immunodominant fraction of the PV self-antigen is present in sera of healthy individuals and patients as a circulating 30 kDa fragment (sDsg3). These findings provide a good explanation for the N-terminal specificity of antibody production and peptide recognition in PV patients by B and T cell, respectively. Moreover, the presence of the sDsg3 in human sera could allow to reconsider pemphigus as a disease against a circulating antigen; once produced, PV-autoantibodies also recognize the 130 kDa epidermal antigen desmoglein 3 on keratinocyte surface (kDsg3), thus triggering the acantholysis and the clinical manifestations of pemphigus. Copyright © by Biolife.