Molecular detection of Chlamydia abortus in a stranded Mediterranean striped dolphin Stenella coeruleoalba.

This study reports gross, histopathological, and molecular features of a Chlamydia abortus infection in a stranded female striped dolphin Stenella coeruleoalba from the Tyrrhenian coast of southern Italy. Post-mortem examination revealed liver congestion, splenic lymphoid depletion with capsular petechiae, and pneumonia. Histology revealed disseminated intravascular coagulation with vasculitis and congestion. Hepatocellular and acute myocardial degeneration were also observed. Basophilic, coccobacillary inclusions consistent with Chlamydia spp. were observed histologically in the type II pneumocytes, myocardial fibers, and hepatocytes, and in macrophages and plasma cells of liver, spleen, and prescapular lymph nodes. Chlamydial antigen was detected by immunofluorescence assay using genus-specific anti-Chlamydia antibodies. PCR assay revealed C. abortus in spleen, liver, heart, and lungs. C. abortus was the only pathogen detected. The main pathological changes suggest that Chlamydia infection may have been the cause of stranding and death of the striped dolphin. This case represents the first molecular detection of a member of the Chlamydiaceae in a marine mammal.

Modeling the modified drug release from curved shape drug delivery systems - Dome Matrix®.

The controlled drug release from hydrogel-based drug delivery systems is a topic of large interest for research in pharmacology. The mathematical modeling of the behavior of these systems is a tool of emerging relevance, since the simulations can be of use in the design of novel systems, in particular for complex shaped tablets. In this work a model, previously developed, was applied to complex-shaped oral drug delivery systems based on hydrogels (Dome Matrix®). Furthermore, the model was successfully adopted in the description of drug release from partially accessible Dome Matrix® systems (systems with some surfaces coated). In these simulations, the erosion rate was used asa fitting parameter, and its dependence upon the surface area/volume ratio and upon the local fluid dynamics was discussed. The model parameters were determined by comparison with the drug release profile from a cylindrical tablet, then the model was successfully used for the prediction of the drug release from a Dome Matrix® system, for simple module configuration and for module assembled (void and piled) configurations. It was also demonstrated that, given the same initial S/V ratio, the drug release is independent upon the shape of the tablets but it is only influenced by the S/V evolution. The model reveals itself able to describe the observed phenomena, and thus it can be of use for the design of oral drug delivery systems, even if complex shaped.

Pathology and molecular analysis of Hapalotrema mistroides (Digenea: Spirorchiidae) infecting a Mediterranean loggerhead turtle Caretta caretta.

Turtle blood flukes belonging to the family Spirorchiidae (Digenea) represent a major threat for sea turtle health and are considered the most important parasitic cause of turtle stranding and mortality worldwide. Despite the large diversity of spirorchiid species found globally, there are only 2 records for free-ranging Mediterranean sea turtles that date back to the late 1800s involving just Hapalotrema mistroides Monticelli, 1896. This study describes the first fatal confirmed case of spirorchiidiasis in a free-ranging Mediterranean loggerhead turtle Caretta caretta (Linnaeus) and, owing to the complexities of taxonomic identification of these parasites, provides the first molecular characterization and phylogenetic analysis of H. mistroides from the Mediterranean Sea. The loggerhead turtle showed cachexia and digestive disorders associated with severe damage to the pancreas and intestinal ganglia, caused by deposition of Hapalotrema eggs forming granulomas. Massive Hapalotrema egg emboli in several tissues and organs and encephalitis were the most probable contributions to the death of the turtle. The congruence between the phylogenetic analysis of both the ITS2 and 28S rDNA resolved the Italian and USA H. mistroides as the same species, confirming the parasite identification. The case here described clearly indicates that the blood flukes should be considered in the differential diagnosis of Mediterranean sea turtle diseases.

Possible role of oral ibandronate administration in Osteonecrosis of the Jaw: a case report.

We describe a case of Osteonecrosis of the Jaw (ONJ) that developed in a 65-year-old Caucasian woman with osteopenia and other risk factors who was receiving low doses of oral bisphosphonate therapy (ibandronate, 150 mg monthly). Computed tomography (CT), panoramic radiographs (OPT), 99mTc-Sn-MDP scintigraphy, and magnetic resonance imaging (MRI) were performed to study the diseased area; cytological examination also revealed the presence of suppurative material around the area of exposed bone. A diagnosis of bisphosphonate-related osteonecrosis of the jaw complicated by osteomyelitis was made. The patient was prescribed a drug protocol consisting of metronidazole 250 mg 2 times daily, chlorhexidine mouthwashes 3 times daily and chewing exercises for two months. Ibandronate was stopped and replaced with strontium ranelate. The symptoms improved and the patient is still under close follow-up. Assessment of the benefits versus risks is particularly necessary in patients with several risk factors to ascertain their eligibility for treatment with antiresorptive drugs and when this is not possible to choose alternative medications.

Sjögren's syndrome pathological neovascularization is regulated by VEGF-A-stimulated TACE-dependent crosstalk between VEGFR2 and NF-κB.

We explore the involvement of tumor necrosis factor α (TNF-α)-converting enzyme (TACE) in vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR2) (VEGF-A/VEGFR2)-mediated angiogenesis in Sjögren's syndrome (SS), one of the most common autoimmune rheumatic diseases. To test the hypothesis that SS autoantibodies (Abs) regulate VEGF-A/VEGFR2 expression by a TACE-dependent nuclear factor-κB (NF-κB) activation pathway, their effects on the expression and activation of the VEGF-A/TACE/VEGFR2/NF-κB pathway were determined in human salivary gland epithelial cells (SGEC). An enhanced angiogenesis in SS salivary gland biopsies was observed, associated with an increased VEGF-A expression and activation of VEGF-A/VEGFR2 signaling. Human cytokine array analysis of the pro-inflammatory and pro-angiogenic protein response in SGEC treated with SS Abs revealed an overexpression of multiple pro-angiogenic factors. TACE RNA knockdown, the use of anti-VEGF-A monoclonal antibody and the inhibition of NF-κB activity significantly abrogated the release of pro-angiogenic factors, demonstrating that VEGF-A/TACE/VEGFR2/NF-κB axis dysfunction may be contributory to pathogenesis and exacerbation of this autoimmune condition.

Use of microwaves for in-situ removal of pollutant compounds from solid matrices.

Thermal treatments are the most used methods to remediate contaminated solids. However, they may seriously damage the otherwise recoverable matrices, especially when mild operating conditions cannot be used. Microwaves recently raised as a powerful tool in industrial engineering for their ability, among other advantages, to offer a selected heating, thus allowing to treat and remove only the undesired components of a matrix. This work approaches the microwave assisted thermal treatments of waste from a physical-chemical point of view. Two recovering operations have been performed, respectively, on a soil contaminated by volatile organic compounds and on a ceramic filter spoiled by soot, using two specially designed prototypes, both realized on pre-pilot scale. The heat and mass transfer balances have then been analyzed in their more general form, and terms related to the use of microwaves outlined. Solutions of the differential equations have been applied to interpret the effects of microwaves on rate and efficiency of the remediation processes.

A quantum mechanical study of TiCl3 alpha, beta and gamma crystal phases: geometry, electronic structure and magnetism.

The electronic structure of different magnetic states of alpha, beta and gamma modifications of TiCl(3) has been computed employing the density functional theory with periodic boundary conditions and localized Gaussian basis sets. The analysis of the density of the electronic states (DOS) and of the spin density makes it possible to classify these halides as Mott-Hubbard insulators, where the band gap appears a result of large on-site Coulomb interaction. For each crystalline phase, the relative stability of different magnetic states has been analyzed in terms of exchange mechanisms. The electronic population data along with the spin density maps support the assumption of a d(1) Titanium ion in a distorted octahedral crystal field, notwithstanding the not fully ionic character of TiCl(3) modifications. Dispersion forces are particularly important for this material: a classical correction (of the type f(R)/R(6)) has been added to the DFT energies and gradients, providing a good agreement with structural data.

Osteoprotegerin and RANKL in the pathogenesis of osteoporosis in patients with thalassaemia major.

Osteoporosis represents an important cause of morbidity in thalassaemia major patients; the etiopathogenesis is multifactorial and includes expansion of the bone marrow, endocrine disorders, iron overload and genetic factors. Two cytokines, osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL), have recently been identified as important mediators in the pathogenesis of osteoporosis. In this study, the possible role of the OPG-RANKL system in the pathogenesis of osteoporosis in thalassemia major is assessed, as well as any correlations between the serum levels of OPG and RANKL and bone mineral density (BMD), 17 beta-estradiol and free testosterone and the relationship between T-score of BMD and OPG/RANKL ratio. In 31 thalassaemia major patients and a control group, the serum values of OPG and RANKL were assayed and correlated with BMD, as well as with the sex hormones values. All the thalassemic patients had reduced BMD and 35.5% presented osteoporosis. The thalassemic patients had significantly higher serum levels of OPG than the controls, while their higher RANKL levels, were at the threshold of significance. The OPG/RANKL ratio showed higher level respect to the controls. No statistically significant correlation was observed between the T-score and RANKL neither between the T-score and OPG nor between T-score and OPG/RANKL ratio. Instead, a statistically significant correlation was found between the T-score and free testosterone and between the T-score and 17 beta-estradiol. There was no correlation between the sex hormones and OPG and RANKL. The increased OPG values in thalassemic patients could be considered to compensate the increased bone turnover. The authors confirm hypogonadism as a primary etiopathogenetic factor in the reduced BMD observed in thalassaemia major patients.

Anti-Ro/SSA autoantibody-mediated regulation of extracellular matrix fibulins in human epithelial cells of the salivary gland.

OBJECTIVES: The fibulins are a family of extracellular matrix (ECM) molecules that regulate the organ shape along with other growth factors and stromal cells and have recently been shown to be involved in a variety of cellular functions including proliferation, migration, differentiation, and survival. Important changes in acinar and ductal morphology and function, together with pronounced ECM remodelling, are detectable in the labial salivary glands (LSGs) of patients with Sjögren's syndrome (SS). Here we report the in vitro expression of the recently identified ECM proteins fibulin-6 and fibulin-7 by human salivary gland epithelial cells (SGECs). The ability of anti-Ro/SSA autoantibodies (Abs) to modulate fibulin-6 and fibulin-7 expression was investigated. METHODS: Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR were used to analyse fibulin-6 and fibulin-7 mRNA expression. Confocal microscopy and fluorescence-activated cell sorting (FACS) were used to study expression of the proteins in primary human SGEC cultures, established from biopsies of minor LSGs, in both untreated control cells and anti-Ro/SSA Abs-treated cells. RESULTS: The methods used show the expression of fibulin-6 and fibulin-7 in SGECs. Treatment of cells with anti-Ro/SSA Abs results in a down-regulation of fibulin-6 mRNA expression whereas no significant differences were observed in fibulin-7 expression between untreated and treated cells. CONCLUSION: Dysregulation of fibulin expression in SGECs by anti-Ro/SSA Abs may contribute to disorganization of the ECM environment and thus cause injury to the salivary gland architecture and functionality observed in SS.

Hashimoto's thyroiditis in Melkersson-Rosenthal syndrome patient: casual association or related diseases?

Melkersson-Rosenthal syndrome (MRS) is a rare disorder of unknown etiology. MRS is classically defined as a triad of recurrent orofacial edema, relapsing paralysis of the facial nerve, and fissured tongue. The authors present the case of a 52-year-old woman with orofacial swelling and facial pain attacks. The patient reported to suffer of hypothyroidism and laboratory findings disclosed free triiodothyronine (FT(3)), free thyroxine (FT(4)), and thyrotropin (TSH) altered. Endocrinological consult led to the diagnosis of Hashimoto's thyroiditis. Antithyroper-oxidase antibodies (anti-TPO) were highly elevated and thyroid function tests had evidenced a clinically significant hypothyroidism. A link between MRS and immunological disorders such as sarcoidosis, Crohn's disease, unilateral anterior uveitis and multiple sclerosis was documented. The literature did not report any association between Hashimoto's thyroiditis and Melkersson-Rosenthal syndrome. The presence of the anti-TPO antibodies in the case reported here could suggest a possible correlation between immunological alteration characteristic of autoimmune thyroiditis and MRS.

[Hospital anxiety and depression]. / Ansia e depressione nelle ospedalizzazioni.

OBJECTIVE: The study investigate the presence of depressive disorders in patient who are taken in general hospital, to prevent and reduce the risk of developing a psychological pathology (anxious-depressive symptoms). METHODS: We used two tests: 1. General Health Questionnaire (GHQ-12) by Goldberg, a self-report questionnaire, consisting of 12 items, used to estimate the probability of detecting non-psychotic mental disorders and problems in every day's activity. 2. Personal Health Questionnaire (PHQ), a self-report questionnaire, consisting of 10 items. It is a new tool for detecting individuals with ICD-10 depressive disorders, used to estimate the probability to individuate the presence of major depression symptoms, and minor depression symptoms. Tests have been administered to 140 subjects (males' experimental group, 32 subjects; males' control group, 20 subjects; females' experimental group 58 subjects; females' control group, 30 subjects) from eight departments of university hospital, medical faculty, and social services. RESULTS: It can be noted that with GHQ-12, the presence of non-psychotic mental disorders does not come out: just the females' group, experimental and control, shows some problems with insomnia and stress. On the other hand, with PHQ, the presence of no great entity depressive symptoms comes out for all groups. In the experimental groups the quantity of depressive symptoms is greater than in the control groups. CONCLUSIONS: We have find the presence of a number of depressive symptoms into a hospitalized population. It is known that detecting such symptoms is important for protection and care of depressive disorders in hospitalized and non-hospitalized populations.

Siögren's syndrome: anti-Ro and anti-La autoantibodies trigger apoptotic mechanism in the human salivary gland cell line, A-253.

AIM: The Sjögren's syndrome (SS) is an autoimmune rheumatic disease that targets salivary and lacrimal glands, characterized by a high concentration of autoantibodies in the serum. The anti-Ro and anti-La autoantibodies are present in approximately 70-90% of the patients with primary SS and this presence is correlated to extraglandular manifestations. The objective of this work was to explore the cellular apoptotic pathway triggered by binding and penetration of anti-Ro and anti-La autoantibodies, isolated from the total IgG fraction of patients with primary SS, in the human salivary gland cell line A-253. METHODS: The sera were obtained from 13 healthy volunteers and 13 patients with primary SS. The IgG was obtained from sera through precipitation with ammonium sulfate and the anti-La and anti-Ro autoantibodies were purified using Sepharose 4B-Ro and Sepharose 4B-La affinity columns. The methods used to evaluate the apoptosis were: DNA fragmentation, immunofluorescence and immunoenzymatic tests. RESULTS: In the salivary gland cells, the anti-Ro and anti-La autoantibodies: 1) are able to penetrate; 2) induce DNA fragmentation and cleavage and activation of the effector caspase-3. In the same experimental condition, IgG purified from healthy sera did not have any apoptotic effect on the human salivary gland cell line. CONCLUSION: Anti-Ro and anti-La autoantibodies mediate the apoptosis the human salivary gland cells A-253 in a caspase-3 dependent manner.

Anti-Ro and anti-La autoantibodies induce TNF-alpha production by human salivary gland cells: an in vitro study.

In this report, we demonstrate that both TNFR1 and the TNFR2 are expressed on the salivary gland cell line A-253 cell membrane. Furthermore, cell treatment with anti-Ro and anti-La autoantibodies from Sjögren IgG determined TNF-alpha production, clarifying which could be the inducer of the extrinsic pathway of apoptosis in salivary gland cells.

Autoantibodies from Sjögren's syndrome induce activation of both the intrinsic and extrinsic apoptotic pathways in human salivary gland cell line A-253.

Sjögren's syndrome (SS) is an autoimmune rheumatic disease that targets salivary and lachrymal glands, characterized by a high concentration of serum autoantibodies directed against nuclear and cytoplasmic antigens. It is known that autoantibodies can enter viable cells and this phenomenon has functional consequences including activation of apoptotic process. The objective of this work was to explore whether autoantibodies contained in IgG purified from Sjögren sera trigger apoptotic process in an experimental model represented by the human salivary gland cell line A-253. To define if the intrinsic or extrinsic pathways are activated, we examined which caspases are critical for inducing cell death. The results have demonstrated that morphological changes and DNA laddering, consistent with apoptotic cell death, occurred in A-253 cells treated with IgG from Sjögren sera. Sjögren IgG induced cleavage and activation of the effector caspase-3 and degradation of the caspase-3 substrate poly(ADP-ribose)polymerase. Both the intrinsic and extrinsic apoptotic pathways were activated, since both caspase-8 and caspase-9 cleavages occurred. In conclusion, autoantibodies contained in IgG purified from Sjögren sera mediate apoptosis of the A-253 cell line in a caspase-dependent manner.

Study on the effectiveness of a nifedipine gel for treatment of Raynaud?s phenomenon.

The aim of this study is to evaluate the efficacy of a nifedipine gel in patients with primary or secondary Raynaud?s phenomenon. Photopletismography was the instrumental examination test used to evaluate recovery time (time necessary for recuperation of normal capillary circulation) in 17 patients with primary or secondary Raynaud?s phenomenon before and after the application of the gel. It emerged that of the 17 patients who used the gel, in 3 cases the recovery time was reduced, in 9 cases the recovery time was cancelled (no spasm occurred), in 5 cases the recovery time was not modified. Therefore, in more than 70 percent of patients the drug had a positive effect. Besides, 50 percent of the patients referred an improvement of the subjective symptomatology with reduction of cooling, torpidity, ache and paresthesias of the fingers. The results obtained, even if related to a restricted number of patients and to a brief interval of time, show the effectiveness of this drug in patients with primary or secondary Raynaud?s phenomenon. We believe that these results, presented here for the first time, are important for investigators involved in the study of Raynaud?s disease.

Receptor activator of NF(Kappa)B ligand/osteoprotegerin (RANKL/OPG) system and osteopontin (OPN) serum levels in a population of apulian postmenopausal women.

AIM: Osteoporosis is a bone disease, characterized by a reduction of bone resistance; in postmenopausal period bone metabolism is imbalanced. Several parameters have been proposed as markers of bone metabolism; the attention have been recently placed on the receptor of activator of NF(Kappa)B ligand receptor (RANKL) and osteoprotegerin (OPG), namely RANKL/OPG system. The aim of this paper is to evaluate changes in postmenopausal women in serum concentration of OPG, RANKL, and their ratio (i.e. RANKL/OPG ratio), osteopontin (OPN), bone-type alcaline phosphatase (BAP), serum-N-Telopeptide of type I collagen (serum-NTX); and their correlations with bone mineral density (BMD). METHODS: An Apulian population group of 163 native postmenopausal women were followed at the Osteoporosis Centre of Policlinico of Bari (Southenrn Italy). Patients were classified into 3 separate groups, according to T-score: osteoporotic, osteopenic and control group. Serum concentrations of OPG, RANKL, RANKL/OPG ratio, BAP and NTX have been calculated. RESULTS: No differences were found in OPG and BAP values. Significant correlations were found in the osteopenic group between OPG and RANKL (negative), and between RANKL and OPN or serum-NTX, OPN and serum-NTX or RANKL/OPG ratio, BAP and serum-NTX, serum-NTX and RANKL/OPG ratio (positive). In the other groups a significant correlation was observed between BAP and NTX. CONCLUSIONS: In postmenopausal women, important modifications of bone metabolism markers (i.e. RANKL, OPG and OPN) could be due to serious engagement of bone turnover, especially in the pre-osteoporotic phase. Low bone density in postmenopausal women should be identified as soon as possible, and urgent measures are needed to reverse the process. Parameters namely RANKL e OPG may become an important index for the evaluation of the activity of drugs against osteoporosis, old and new like AMG 162 (anti-RANKL action).

[The strange case of a patient affected by acromegaly with osteoporomalacia without hypogonadism]. / Lo strano caso di un paziente affetto da acromegalia con osteoporomalacia senza ipogonadismo.

Acromegaly is a rare disease that, in the majority of cases, is due to the presence of a benign growth hormone (GH)-producing tumor of the pituitary. Growth hormone has profound effects on linear bone growth, bone metabolism, and bone mass. In acromegaly, the skeletal effects of chronic GH excess have been mainly addressed by evaluating bone mineral density (BMD). Most data were obtained in patients with active acromegaly, and apparently high or normal BMD was observed in the absence of hypogonadism. The Autors describe a case of patient affected by acromegaly without hypogonadism with serious osteoporosis and biological signs of osteomalacia.