Ultrastructure study of skin fibroblasts in patients with Ehlers-Danlos Syndrome (EDS): preliminary results.

AIM OF THE STUDY: To investigate, in vivo and in vitro, the fibroblast-to-myofibroblast transition in patients with hypermobile Ehlers-Danlos Syndrome (EDS). To analyze the dermis of patients with classical form of EDS (cEDS) and with hEDS, to identify qualitative and/or quantitative differences in ECM component and ultrastructural changes in collagen. MATERIALS AND METHODS: Seven subjects, aged over 18, two with cEDS and five with hEDS underwent two skin biopsy. One sample was prepared for transmission electron microscopy (TEM), the other for immunofluorescence. The diameter of collagen fibers was measured with TEM. Fibrils were analyzed in four patients: the two with cEDS and two with hEDS. For each patient, the diameter of n=250 collagen fibrils was measured. αSMA was used as specific marker for myofibroblast to highlight their presence in vivo in the skin of patients with hEDS. RESULT: IF observation could not assess an increased expression of αSMA in hEDS patients, which showed no statistical difference compared to classic form patients. The major result from the analysis of TEM images is the clear difference in ECM composition between the two forms of EDS: ECM in hEDS is optically more dense and more prominently composed of elastic fibers. CONCLUSION: Our study provides the following important evidence: 1) the absence in vivo of dermal fibroblasts in patients with hEDS, demonstrated by αSMA negativity; 2) the presence of statistically significant changes in the diameter of collagen fibrils between the classic and the hypermobile forms.

Clinical and biochemical response to neridronate treatment in a patient with osteoporosis-pseudoglioma syndrome (OPPG).

Osteoporosis-pseudoglioma syndrome (OPPG) is a rare autosomal recessive syndrome characterized by juvenile-onset osteoporosis and ocular abnormalities due to a low-density lipoprotein receptor-related protein 5 (LRP5) gene mutation. Treatment with bisphosphonates, particularly with pamidronate and risedronate, has been reported to be of some efficacy in this condition. We report on a patient with OPPG due to an LRP5 gene mutation, who showed an encouraging response after a 36-month period of neridronate therapy. We report a case of a patient treated with bisphosphonates. Bisphosphonates should be administered in OPPG patients as a first-line therapy during early childhood.

Serum creatine kinase isoenzymes in children with osteogenesis imperfecta.

This study evaluates serum creatine kinase isoenzyme activity in children with osteogenesis imperfecta to determine its usefulness as a biochemical marker during treatment with bisphosphonate. The changes of creatine kinase (CK) isoenzyme activity during and after discontinuation therapy were observed. These results could be useful in addressing over-treatment risk prevention. INTRODUCTION: The brain isoenzyme of creatine kinase (CKbb) is highly expressed in mature osteoclasts during osteoclastogenesis, thus plays an important role in bone resorption. We previously identified high serum CKbb levels in 18 children with osteogenesis imperfect (OI) type 1 treated for 1 year with bisphosphonate (neridronate). In the present study, serum CK isoenzymes were evaluated in the same children with continuous versus discontinued neridronate treatment over a further 2-year follow-up period. METHODS: This study included 18 children with OI type 1, 12 with continued (group A) and 6 with ceased (group B) neridronate treatment. Auxological data, serum biochemical markers of bone metabolism, bone mineral density z-score, and serum total CK and isoenzyme activities were determined in both groups. RESULTS: Serum CKbb was progressively and significantly increased in group A (p < 0.004) but rapidly decreased to undetectable levels in group B. In both groups, the cardiac muscle creatine kinase isoenzyme (CKmb) showed a marked decrease, while serum C-terminal telopeptide (CTx) levels were almost unchanged. CONCLUSIONS: This study provides evidence of the cumulative effect of neridronate administration in increasing serum CKbb levels and the reversible effect after its discontinuation. This approach could be employed for verifying the usefulness of serum CKbb as a biochemical marker in patients receiving prolonged bisphosphonate treatment. Moreover, the decreased serum CKmb levels suggest a systemic effect of these drugs.

Cutaneous toxoplasmosis in an immunosuppressed dog / Toxoplasmose cutânea em um cão imunosuprimido

A seven-year-old female spayed Schnauzer was presented with cutaneous ulcerated nodular lesions shortly after the beginning of an immunosuppressive treatment for immune-mediated hemolytic disease. Cytology was performed and a great number of neutrophils and banana-shaped organisms were observed. Biopsy showed a neutrophilic and histiocytic dermatitis and panniculitis with myriads of intralesional bradyzoites cysts and tachyzoites. PCR analysis was positive for Toxoplasma gondii and negative for Neospora caninum. Immunohistochemistry confirmed intralesional T. gondii antigens. This study reports a rare case of cutaneous toxoplasmosis in an immunosuppressed dog...

Uma cadela Schnauzer, castrada, de sete anos apresentou lesões cutâneas nodulares ulceradas pouco tempo após início de tratamento imunossupressor para doença hemolítica imunomediada. Foi realizado exame citológico, e um grande número de neutrófilos e estruturas em forma de banana foi observado. Biópsia mostrou dermatite e paniculite neutrofílica e histiocítica com miríades de taquizoítos e cistos de bradizoítos intralesionais. PCR foi positivo para Toxoplasma gondii e negativo para Neospora caninum. Imuno-histoquímica confirmou antígenos de T. gondii intralesionais. Este trabalho relata um caso raro de toxoplasmose cutânea em um cão imunossuprimido...

Radioimmunological detection of anti-transglutaminase autoantibodies in human saliva: a useful test to monitor coeliac disease follow-up.

BACKGROUND: Serum radioimmunoassay (RIA) tissue transglutaminase autoantibodies (tTG-Abs) proved to be a sensitive test also during coeliac disease (CD) follow-up. We demonstrated that RIA tTG-Abs could be detected in human saliva. AIM: To evaluate salivary RIA tTG-Abs in coeliac children on gluten-free diet (GFD). METHODS: Saliva and serum samples from 109 coeliac children were evaluated at diagnosis (group 1: 71 females, median age 9.4 years) and 58 of them on GFD: 36 after 3-6 months (group 2a), 34 at 9 months or more (group 2b). Two gender- and age-matched control groups: 89 gastroenterological patients (group 3) and 49 healthy subjects (group 4) participated in the study. Saliva and serum tTG-Abs were detected by RIA and compared with serum tTG-Abs ELISA and IgA anti-endomysium antibodies (EMA). RESULTS: Salivary RIA tTG-Abs were found in 94.5%, 66.7% and 50.0% of groups 1, 2a and 2b CD patients and in 98.2%, 72.2% and 50.0% of corresponding serum samples, respectively. tTG-Abs decreased with GFD progression and a correlation was found between saliva and serum titres (r = 0.75, P = 0.0001). During the CD follow-up, salivary and serum RIA sensitivities were comparable, and higher with respect to EMA and ELISA. CONCLUSIONS: This study demonstrates that it is possible to detect salivary tTG-Abs with high sensitivity not only at CD diagnosis, but also during GFD.

Gastric emptying time in cyclic vomiting syndrome in children.

Pathogenesis of cyclic vomiting syndrome (CVS) is still uncertain. Aim of our study has been to verify the role of gastric emptying time in children affected by CVS. We studied 9 children with CVS who underwent measurement of gastric emptying time by real-time ultrasonography after administration of a mixed, solid-liquid meal. In all the children gastric emptying time resulted in normal range. Our data suggest that mobility abnormalities of the stomach don't play a key role in the pathogenesis of CVS.

Dominant oncogenes, tumor suppressors, and radiosensitivity.

A variety of conflicting results appeared in the literature concerning the effect of dominant oncogenes on the sensitivity to irradiation and to anticancer agents in a number of cell lines of human and animal origin. In this report we provide evidence supporting the hypothesis that the tumor suppressor gene p53 and the apoptosis suppressor gene bcl2 modulate the effect of dominant oncogenes and that the effect of dominant oncogenes on resistance or sensitivity is dependent on the balance between the expression of p53 and bcl2.

Bcl-2 overexpression abolishes early calcium waving preceding apoptosis in NIH-3T3 murine fibroblasts.

Overexpression of the apoptosis protection gene bcl-2 abolished the earliest response to serum withdrawal in NIH-3T3 murine fibroblasts, i.e., the abrupt cytoplasmic free calcium drop. This phenomenon, also observed in a myeloid cell line, led us to propose this ionic waving as an early apoptotic signal. Its abolition by bcl-2 overexpression suggests that this gene plays a role also on early events "priming" apoptosis.

Apoptosis induction in 32D cells by IL-3 withdrawal is preceded by a drop in the intracellular calcium level.

The 32D murine myeloid cell line is dependent on interleukin-3 (IL-3) for growth in vitro. We show here that IL-3 induces a slow increase in endocellular calcium, which is sizeable two hours after the addition. Withdrawal of the cytokine induces apoptosis, whose classic late events are evident after 16/18 hours, and are preceded by a calcium drop during the first 2/3 hours after IL-3 subtraction. Calcium drop is here proposed as a trigger of the apoptotic process, in agreement with other recently reported findings.