Localized delivery of fibroblast growth factor-2 and brain-derived neurotrophic factor reduces spontaneous seizures in an epilepsy model.

A loss of neurons is observed in the hippocampus of many patients with epilepsies of temporal lobe origin. It has been hypothesized that damage limitation or repair, for example using neurotrophic factors (NTFs), may prevent the transformation of a normal tissue into epileptic (epileptogenesis). Here, we used viral vectors to locally supplement two NTFs, fibroblast growth factor-2 (FGF-2) and brain-derived neurotrophic factor (BDNF), when epileptogenic damage was already in place. These vectors were first characterized in vitro, where they increased proliferation of neural progenitors and favored their differentiation into neurons, and they were then tested in a model of status epilepticus-induced neurodegeneration and epileptogenesis. When injected in a lesioned hippocampus, FGF-2/BDNF expressing vectors increased neuronogenesis, embanked neuronal damage, and reduced epileptogenesis. It is concluded that reduction of damage reduces epileptogenesis and that supplementing specific NTFs in lesion areas represents a new approach to the therapy of neuronal damage and of its consequences.

Localized overexpression of FGF-2 and BDNF in hippocampus reduces mossy fiber sprouting and spontaneous seizures up to 4 weeks after pilocarpine-induced status epilepticus.

PURPOSE: We have recently reported that viral vector-mediated supplementation of fibroblast growth factor-2 (FGF-2) and brain-derived neurotrophic factor (BDNF) in a lesioned, epileptogenic rat hippocampus limits neuronal damage, favors neurogenesis, and reduces spontaneous recurrent seizures. To test if this treatment can also prevent hippocampal circuit reorganization, we examined here its effect on mossy fiber sprouting, the best studied form of axonal plasticity in epilepsy. METHODS: A herpes-based vector expressing FGF-2 and BDNF was injected into the rat hippocampus 3 days after an epileptogenic insult (pilocarpine-induced status epilepticus). Continuous video-electroencephalography (EEG) monitoring was initiated 7 days after status epilepticus, and animals were sacrificed at 28 days for analysis of cell loss (measured using NeuN immunofluorescence) and mossy fiber sprouting (measured using dynorphin A immunohistochemistry). KEY FINDINGS: The vector expressing FGF-2 and BDNF decreased both mossy fiber sprouting and the frequency and severity of spontaneous seizures. The effect on sprouting correlated strictly with the cell loss in the terminal fields of physiologic mossy fiber innervation (mossy cells in the dentate gyrus hilus and CA3 pyramidal neurons). SIGNIFICANCE: These data suggest that the supplementation of FGF-2 and BDNF in an epileptogenic hippocampus may prevent epileptogenesis by decreasing neuronal loss and mossy fiber sprouting, that is, reducing some forms of circuit reorganization.

Hippocampal FGF-2 and BDNF overexpression attenuates epileptogenesis-associated neuroinflammation and reduces spontaneous recurrent seizures.

Under certain experimental conditions, neurotrophic factors may reduce epileptogenesis. We have previously reported that local, intrahippocampal supplementation of fibroblast growth factor-2 (FGF-2) and brain-derived neurotrophic factor (BDNF) increases neurogenesis, reduces neuronal loss, and reduces the occurrence of spontaneous seizures in a model of damage-associated epilepsy. Here, we asked if these possibly anti-epileptogenic effects might involve anti-inflammatory mechanisms. Thus, we used a Herpes-based vector to supplement FGF-2 and BDNF in rat hippocampus after pilocarpine-induced status epilepticus that established an epileptogenic lesion. This model causes intense neuroinflammation, especially in the phase that precedes the occurrence of spontaneous seizures. The supplementation of FGF-2 and BDNF attenuated various parameters of inflammation, including astrocytosis, microcytosis and IL-1ß expression. The effect appeared to be most prominent on IL-1ß, whose expression was almost completely prevented. Further studies will be needed to elucidate the molecular mechanism(s) for these effects, and for that on IL-1ß in particular. Nonetheless, the concept that neurotrophic factors affect neuroinflammation in vivo may be highly relevant for the understanding of the epileptogenic process.

Vulvar melanocytic nevi: a dermoscopic investigation.

BACKGROUND: Very few dermoscopic investigations into pigmented genital lesions have been performed to date. To the best of our knowledge, no dermoscopical description of vulvar melanocytic nevi (MN) has yet been proposed. OBJECTIVE: Our aims were to analyze the dermoscopic features of vulvar MN and to review the literature of dermoscopy of the vulvar area. METHODS: Among 74 women presenting 84 vulvar pigmented lesions, 10 who received a visual diagnosis of MN were submitted to dermoscopic and histopathological analysis. RESULTS: Histology confirmed the clinical diagnosis of MN in 8 out of 10 cases, while the remaining 2 pigmented lesions were diagnosed as seborrheic keratosis. Vulvar MN were generally found to share similar dermoscopic patterns with their cutaneous equivalent. CONCLUSIONS: Although further observation will be necessary to provide additional data and establish key morphological criteria and/or algorithms useful for differentiating between melanocytic and nonmelanocytic vulvar lesions, dermoscopy should be considered an investigative tool, as clinical observation alone may not be sufficient for accurate diagnosis.

Fgf-2 overexpression increases excitability and seizure susceptibility but decreases seizure-induced cell loss.

Fibroblast growth factor 2 (FGF-2) has multiple, pleiotropic effects on the nervous system that include neurogenesis, neuroprotection and neuroplasticity. Thus, alteration in FGF-2 expression patterns may have a profound impact in brain function, both in normal physiology and in pathology. Here, we used FGF-2 transgenic mice (TgFGF2) to study the effects of endogenous FGF-2 overexpression on susceptibility to seizures and to the pathological consequences of seizures. TgFGF2 mice display increased FGF-2 expression in hippocampal pyramidal neurons and dentate granule cells. Increased density of glutamatergic synaptic vesicles was observed in the hippocampus of TgFGF2 mice, and electrophysiological data (input/output curves and patch-clamp recordings in CA1) confirmed an increase in excitatory inputs in CA1, suggesting the presence of a latent hyperexcitability. Indeed, TgFGF2 mice displayed increased susceptibility to kainate-induced seizures compared with wild-type (WT) littermates, in that latency to generalized seizure onset was reduced, whereas behavioral seizure scores and lethality were increased. Finally, WT and TgFGF2 mice with similar seizure scores were used for examining seizure-induced cellular consequences. Neurogenesis and mossy fiber sprouting were not significantly different between the two groups. In contrast, cell damage (assessed with Fluoro-Jade B, silver impregnation and anti-caspase 3 immunohistochemistry) was significantly lower in TgFGF2 mice, especially in the areas of overexpression (CA1 and CA3), indicating reduction of seizure-induced necrosis and apoptosis. These data suggest that FGF-2 may be implicated in seizure susceptibility and in seizure-induced plasticity, exerting different, and apparently contrasting effects: favoring ictogenesis but reducing seizure-induced cell death.

Different production of soluble HLA-G antigens by peripheral blood mononuclear cells in ulcerative colitis and Crohn's disease: a noninvasive diagnostic tool?

BACKGROUND: HLA-G antigens are nonclassical major histocompatibility complex (MHC) class I molecules characterized by tolerogenic and antiinflammatory properties. Recently, a different expression of HLA-G antigens has been observed between intestinal biopsies of ulcerative colitis (UC) and Crohn's disease (CD) patients. These data suggested a functional role for HLA-G molecules in the diseases and proposed the HLA-G modulation as a marker for the diagnosis of UC and CD. The soluble HLA-G antigens (sHLA-G) are circulating molecules mainly produced by activated peripheral blood CD14+ monocytes. METHODS: We tested, by specific enzyme-linked immunosorbent assay (ELISA), the sHLA-G molecule levels in the supernatants of unstimulated and bacterial lipopolysaccharide (LPS)-stimulated cultures of peripheral blood mononuclear cells (PBMC) from 30 healthy subjects, 10 CD, and 18 UC patients. The data were not influenced by treatment or disease activity. RESULTS: The results confirmed a different sHLA-G expression between the diseases, with a spontaneous secretion of sHLA-G in CD patients but not in UC and healthy subjects. Moreover, a lack of sHLA-G antigens has been reported in UC patient cultures after LPS activation but not in healthy subjects and CD patients. The defective sHLA-G production was related to an impaired IL-10 secretion in UC but not in CD. CONCLUSIONS: Overall, these results confirm the presence of a different biological characteristic between CD and UC patients and suggest sHLA-G production by PBMC as a noninvasive diagnostic tool in the early phases of the diseases.

Modeling and remodeling of human extraction sockets.

INTRODUCTION: The available studies on extraction wound repair in humans are affected by significant limitations and have failed to evaluate tissue alterations occurring in all compartments of the hard tissue defect. AIM: To monitor during a 6-month period the healing of human extraction sockets and include a semi-quantitative analysis of tissues and cell populations involved in various stages of the processes of modeling/remodeling. MATERIAL AND METHODS: Twenty-seven biopsies, representative of the early (2-4 weeks, n=10), intermediate (6-8 weeks, n=6), and late phase (12-24 weeks, n=11) of healing, were collected and analysed. RESULTS: Granulation tissue that was present in comparatively large amounts in the early healing phase of socket healing, was in the interval between the early and intermediate observation phase replaced with provisional matrix and woven bone. The density of vascular structures and macrophages slowly decreased from 2 to 4 weeks over time. The presence of osteoblasts peaked at 6-8 weeks and remained almost stable thereafter; a small number of osteoclasts were present in a few specimens at each observation interval. CONCLUSIONS: The present findings demonstrated that great variability exists in man with respect to hard tissue formation within extraction sockets. Thus, whereas a provisional connective tissue consistently forms within the first weeks of healing, the interval during which mineralized bone is laid down is much less predictable.

Tacrolimus 0.1% ointment: is it really effective in plasma cell vulvitis? Report of four cases.

BACKGROUND: Plasma cell vulvitis is a clinically and histologically well-characterized chronic disease that usually relapses after various topical therapies. Considering the inflammatory nature of the disease, the new topical calcineurin inhibitors have been also employed successfully in few cases of Zoon's balanitis, the corresponding male condition. OBJECTIVE: The aim of our study is to evaluate the effectiveness of tacrolimus ointment in a small group of plasma cell vulvitis sufferers. METHODS: 4 women affected by biopsy-proved plasma cell vulvitis were enrolled, after informed consent. The topical drug was applied twice daily for 6 weeks, then tapered on the basis of the clinical results. Symptoms and objective parameters were obtained periodically at the beginning, after 6 weeks and up to the end of the topical treatment. A final biopsy was performed in 3 out of our 4 patients. The follow-up is still ongoing. RESULTS: The comparative analysis of subjective, objective and histopathological data has shown discordant and less encouraging results than those reported for the corresponding male condition. CONCLUSION: At the moment, topical tacrolimus could be considered an alternative treatment for plasma cell vulvitis only in cases resistant to conventional therapies.

GBR and autogenous cortical bone particulate by bone scraper for alveolar ridge augmentation: a 2-case report.

Scientific literature describes autogenous bone as the gold standard among graft materials for alveolar reconstructive procedures. Alveolar ridge augmentation has been clinically achieved with different forms of autogenous bone, including autogenous cortical bone particulate (ACBP). However, few histologic studies demonstrating the biologic potential and healing dynamics following the use of ACBP are currently available. This case report presents 2 patients in whom atrophic edentulous alveolar crests were submitted to a vertical/lateral ridge augmentation prior to implant placement. The technique was performed through the use of a titanium-reinforced expanded polytetrafluoroethylene (e-PTFE) membrane with an ACBP graft obtained from the retromolar region with a specially designed bone scraper. Bone biopsy specimens were harvested at 9 months after graft placement. Analysis of the reconstructed bone revealed bone with a lamellar quality characterized by a mature osteonic structure. Sparse particles of grafted bone were evident in direct contact with the regenerated bone. Marrow spaces showed a normal stromal component with limited grafted particles.

[Primary biliary cirrhosis and autoimmune hepatitis overlap syndrome. An early case]. / Sindrome di overlap tra epatite autoimmune e cirrosi biliare primitiva. Un caso precoce.

A 21-year old asymptomatic woman had accidental report of increased transaminases. Serologic tests were negative, autoimmune profile was positive for anti-nuclear, antimitochondrial antibodies and rheumatoid factor. Histology of the liver biopsy showed severe necro-inflammatory activity both in biliary epithelium and in intralobular area, suggesting primary biliary cirrhosis/autoimmune hepatitis overlap syndrome.

Retrospective histopathologic reevaluation of 18 cases of plasma cell vulvitis.

OBJECTIVE: To determine some common histologic features in a series of plasma cell vulvitis (PCV) cases. STUDY DESIGN: In a retrospective study, 18 histologic sections, previously obtained by vulvar biopsy in women diagnosed as having PCV, were critically reevaluated by the same pathologist. RESULTS: We observed that plasma cells in the dermal infiltrate were frequently present in a high percentage. Hemosiderin deposition and epithelial atrophy were 2 other histologic parameters useful for the diagnosis of PCV. Lozenge-shaped keratinocytes were rarely observed. CONCLUSION: In our series of PCV the percentage of plasma cells seemed to be the most important parameter when making the diagnosis. In particular, when this percentage was > or = 50% it was a sufficient histologic parameter for a diagnosis of PCV. When the percentage was 25-50%, hemosiderin deposition and epithelial atrophy were additional histologic features helpful in diagnosing PCV. Under 25% plasma cells was nonspecific and related to the mucosal site of involvement.

Presence of detectable levels of soluble HLA-G molecules in CSF of relapsing-remitting multiple sclerosis: relationship with CSF soluble HLA-I and IL-10 concentrations and MRI findings.

We have investigated the presence of non-classical soluble HLA-G molecules (sHLA-G) in cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients and the possible relationships between CSF levels of sHLA-G, classical soluble HLA-I (sHLA-I) molecules, IL-10 amounts and Magnetic Resonance Imaging (MRI) findings were evaluated. We studied by ELISA technique the sHLA-I, sHLA-G and IL-10 levels in CSF of 50 relapsing-remitting (RR) MS patients stratified according to clinical and MRI evidence of disease activity. Thirty-six patients with other inflammatory neurological disorders (OIND) and 41 with non-inflammatory neurological disorders (NIND) were used as controls. CSF mean levels were significantly higher in MS and OIND than in NIND for sHLA-I (p<0.001) and in MS than in controls for sHLA-G (p<0.001), with no differences among the various groups for IL-10 mean concentrations. An increase in CSF sHLA-I was found in MS patients with Gd-enhancing lesions (p<0.01), while sHLA-G and IL-10 were more represented in MS patients without lesional activity on MRI scans (p<0.02). In MRI-inactive MS, CSF IL-10 mean concentrations were significantly greater in patients with CSF-detectable levels of sHLA-G than in those without any evidence of CSF sHLA-G expression (p<0.05). Our findings suggest that CSF classical sHLA-I and non-classical sHLA-G levels may modulate MS activity as assessed by MRI acting in opposite directions. The association observed between sHLA-G and IL-10 when Gd-enhancing lesion resolved indicates a potential immunoregulatory role for IL-10 in the control of MS disease activity by shifting the sHLA-I/sHLA-G balance towards sHLA-G response.

Bilateral orbital involvement in Erdheim-Chester disease.

Erdheim-Chester disease is an idiopathic condition characterized by a xanthogranulomatous process infiltrating the bones, lungs, heart, retroperitoneum and other tissues. This condition is often fatal. Ocular findings are rare. The authors report a case of bilateral xanthelasmas and bilateral massive orbital infiltration in a 61-year-old man with severe retroperitoneal fibrosis, renal and cardiovascular problems. The ophthalmic manifestations and differential diagnosis of this peculiar pathologic condition are discussed.

Eosinophilic cystitis and nephrogenic adenoma of the bladder: a rare association of 2 unusual findings in childhood.

Neither eosinophilic cystitis nor nephrogenic adenoma is often diagnosed in children, with few pediatric cases being reported in the literature. Although these maladies share the same predisposing conditions, namely, chronic irritation or injury to the urothelium and lower urinary tract and symptoms such as dysuria, hematuria, and urinary frequency, their concomitance is exceptional. Herein, we report the case of an 8-year-old boy with a previous history of bladder surgery presenting histologically diagnosed eosinophilic cystitis and nephrogenic adenoma.

Sweet's syndrome: a retrospective clinical, histopathological and immunohistochemical analysis of 11 cases.

The aim of this paper is to report our clinical experience of Sweet's syndrome, a severe dermatological disease which may be extremely important to recognize for the early diagnosis of a neoplastic disorder. Eleven patients affected by Sweet's syndrome, treated at the Department of Dermatology, University of Ferrara, Ferrara, Italy, during 1998 to 2004, were evaluated. A retrospective analysis was performed. Data on age, sex distribution, clinical data, histopathological and immunohistochemical findings and therapy were collected. We observed one patient with idiopathic form, 5 patients affected by the para-inflammatory variant and 5 para-neoplastic cases (with haemoproliferative diseases). The cases with the para-inflammatory form were affected by minor infectious manifestations. Prolonged follow-up is necessary to verify that a case of idiopathic variant is not really a paraneoplastic form. Based on immunohistochemical analysis, we cannot exclude that true histiocytes, immunoreactive for CD68/PGM, infiltrate the dermis in Sweet's syndrome lesions.

Primary lacrimal sac B-cell immunoblastic lymphoma simulating an acute dacryocystitis.

The case of a 72-year-old woman with diffuse large B-cell lymphoma of the lacrimal sac is reported. The patient was evaluated for the first time in our department for tearing of the right eye. One month later, a slightly aching mass appeared over the right lacrimal sac. An acute infectious etiology was suspected and antibiotic therapy was given. When she finally presented with a rapidly growing lesion, she underwent echography and computed tomography followed by incisional biopsy. Results of histopathologic and immunohistochemical evaluation showed a primary, diffuse, large B-cell non-Hodgkin lymphoma of the lacrimal sac. This case demonstrates how difficult the clinical diagnosis of tumors of the lacrimal sac may be in the early stages. The clinical signs, usually aspecific, may be misleading and the diagnosis delayed.