Angiotensin II: immunohistochemical study in Sardinian pterygium.

The angiotensin II (Ang II) is the principal effector peptide of the RAS system. It has a pleiotropic effect and, beside its physiological role, it has the property to stimulate angiogenesis and activate multiple signalling pathways related to cell proliferation. The purpose of the study was to determinate the Ang II expression and localization in Sardinian pterygium and normal conjunctiva by immunohistochemistry, and its possible involvement in the development and progression of the disease. Twenty-three pterygiums and eleven normal conjunctiva specimens obtained from Sardinian patients, were processed for paraffin embedding and assessed for the immunohistochemical revelation of Ang II. Significant Ang II expression was identified in pterygium and conjuntica. Particularly, thirteen pterygium specimens (n=13) displayed exclusively moderate to strong nuclear staining; some specimens (n=5) showed exclusively a moderate cytoplasmatic immunoreactivity, and few specimens (n=2) displayed moderate to strong immunoreactivity in both cytoplasm and nucleus. Statistical significance difference in respect of nuclear and cytoplasmatic localization was observed between normal conjunctiva and pterygium (P=0.038).The results showed a predominant intranuclear localization of Ang II in pterygium epithelial cells, in spite of conjunctiva that mainly showed cytoplasmatic localization. In view of these results, we hypothesized a possible gene expression modulator role played by Ang II in pterygium.

Bilateral cavitations of ganglionic eminence: a fetal MR imaging sign of halted brain development.

SUMMARY: Ganglionic eminence is the main transitory proliferative structure of the ventral telencephalon in human fetal brain and it contributes for at least 35% to the population of cortical interneurons; however data on the human GE anomalies are scarce. We report 5 fetal MR imaging observations with bilateral symmetric cavitations in their GE regions resembling an inverted open C shape and separating the GE itself form the deeper parenchyma. Imaging, neuropathology, and follow-up features suggested a malformative origin. All cases had in common characteristics of lissencephaly with agenesis or severe hypoplasia of corpus callosum of probable different genetic basis. From our preliminary observation, it seems that GE cavitations are part of conditions which are also accompanied by severe cerebral structure derangement.

In vivo detection of cortical abnormalities in BCNU-treated rats, model of cortical dysplasia, using manganese-enhanced magnetic resonance imaging.

The 1-3-bis-chloroethyl-nitrosurea (BCNU)-treated rats represent a good model of cortical dysplasia (CD), as proved by the presence of some histological alterations similar to those observed in human CD, including cortical thinning, laminar disorganization, and heterotopia. The cortical cytoarchitectonics of BCNU-treated rats has been widely investigated by means of histological procedures, immunocytochemistry, and in situ hybridization techniques, implying the sacrifice of the animals. With the aim of identifying brain alterations in vivo to have the possibility of performing longitudinal studies, we used both conventional T(2)-weighted magnetic resonance imaging (MRI) and manganese-enhanced MRI (MEMRI). Though the T(2)-weighted MRI showed the gross anatomical landmarks of BCNU-treated rats, only following Mn(2+) administration T(1)-weighted MRI did reveal the brain cytoarchitectonics both of control and BCNU-treated rats. In particular, changes in MEMRI signal depicted the laminar architecture of control rats while BCNU-treated cortex showed no appreciable changes in MEMRI contrast, consistent with their abnormal cortical lamination. Furthermore, in the treated animals MEMRI revealed hyperintense signals corresponding to heterotopia, as shown by the comparison between MEMRI images and Thionin staining and calbindin immunocytochemistry from the same animals. The qualitative findings obtained with MEMRI were semi-quantitatively confirmed by image segmentation of grey matter. Overall, these data show that MEMRI can be used as a non-invasive technique to investigate cortical alterations in animal models of CD in vivo, giving the possibility to perform longitudinal studies, such as electrophysiological recordings or behavioural investigations.

Combined 7-T MRI and histopathologic study of normal and dysplastic samples from patients with TLE.

OBJECTIVES: The purpose of the study was to investigate the abnormalities of cortical lamination observed in temporal lobe specimens obtained during surgery for intractable temporal lobe epilepsy (TLE) with hippocampal sclerosis. Specifically, we aimed to 1) correlate high-field ex vivo MRI with histopathologic analysis and 2) evaluate the effect of tissue fixation on image contrast. METHODS: A cohort of 13 specimens was considered. T2-weighted imaging and relaxometry were performed during and after fixation using a 7-T experimental scanner. After imaging, the specimens were studied with histopathologic (Black Gold myelin fiber staining) and immunohistochemical (NeuN neuronal staining) methods in order to explore the correspondence between MRI and histopathologic features. RESULTS: The principal findings of this study are that 1) superior MRI contrast is obtained among the cortical layers using completely fixed specimens as opposed to recently excised tissue, 2) the intensity of the T2-weighted MRI signal is lowest (hypointensity) at the site of highest fiber concentration and cellular density, and highest (hyperintensity) when the density of fibers and cells is lowest, and 3) the MRI signal is altered in presence of abnormal cortical lamination (focal cortical dysplasia type IA). CONCLUSIONS: High resolution ex vivo MRI enables the study of intracortical organization in normal and pathologic areas. Comparisons between MRI, NeuN, and Black Gold indicate that the differences apparent in T2-weighted images are mainly related to fiber concentration, although neuronal density might also play a role.

Reliability of the first-trimester cardiac scan by ultrasound-trained obstetricians with high-frequency transabdominal probes in fetuses with increased nuchal translucency.

OBJECTIVE: To examine prospectively the reliability of ultrasound-trained obstetricians performing a first-trimester fetal cardiac scan with high-frequency transabdominal probes, by confirming normal or abnormal heart anatomy, in pregnancies referred for increased nuchal translucency thickness (NT). METHODS: Trained obstetric operators assessed the fetal heart in 133 fetuses with increased NT (> 95th centile) at 11-14 weeks of gestation. A high-frequency transabdominal probe was used to confirm or refute normal cardiac anatomy rather than to establish a specific diagnosis. Following this preliminary screening by the ultrasound-trained obstetrician, specialized fetal echocardiographers rescanned the fetal heart in order to confirm the accuracy of the obstetric operators' findings and to establish a diagnosis in abnormal cases. Fetal cardiologists repeated the examinations at 20 and 32 weeks of pregnancy. Postnatal follow-up lasted 2 years. Twelve fetuses with normal karyotype and normal anatomy were lost to follow-up. RESULTS: A total of 121 fetuses with increased NT between 11 and 14 weeks' gestation were studied. Congenital heart disease (CHD) was detected in 20/121 (16.5%) fetuses. In addition, there were three with mild ventricular disproportion, the right ventricle being larger than the left, considered as a minor non-specific cardiac abnormality. CHD was associated with chromosomal anomalies in 12/20 (60%) cases. Among the 121 fetuses, there was agreement between ultrasound-trained obstetricians and fetal cardiologists in 116 (95.9%) of the cases, and the ultrasound-trained obstetricians correctly identified 18 cases with major cardiac defects. However, there was disagreement in five cases: two with small ventricular septal defects and three with ventricular disproportion. CONCLUSIONS: Our results provide evidence that obstetricians, trained to study the heart in the second trimester, can also differentiate reliably between normal and abnormal heart findings in the first trimester, when using a high-frequency transabdominal ultrasound probe.

Finding of conjunctival melanocytic pigmented lesions within pterygium.

AIMS: Conjunctival pigmented lesions have characteristic clinical and histopathological appearances. Melanocytic pigmented lesions commonly occur in the conjunctiva, although they have not been previously reported in pterygium, a common lesion which originates from conjunctiva. Our aim was to evaluate the possibility of an association between pterygium and conjunctival melanocytic pigmented lesions. METHODS AND RESULTS: A total of 80 samples of pterygium excised from Ecuadorian patients in 2002 were collected. Clinical data were available regarding age, sex, race and place of residence. Histological sections were evaluated for the presence of melanocytic pigmented lesions. Nine cases of conjunctival melanocytic, pigmented lesions within pterygium were found and were classified according to the histopathological criteria previously published for pigmented lesions of the conjunctiva, as naevi and primary acquired melanosis (PAM) with varying degrees of atypia. Five of the nine cases showed primary acquired melanosis without atypia, while two cases had atypia; one case showed features of compound naevus and one lesion was designated as subepithelial naevus. CONCLUSIONS: Our findings suggest that conjunctival melanocytic, pigmented lesions occasionally occur in pterygium. All surgically removed pterygia should undergo careful histopathological examination.

Multiexponential T2-relaxation analysis in cerebrally damaged rats in the absence and presence of a gadolinium contrast agent.

An analysis of the multiexponential relaxation of transverse nuclear magnetization with and without a gadolinium-based paramagnetic contrast agent in spontaneously hypertensive stroke-prone rats (SHR-SP) and in the rat model of ischemia induced by middle cerebral artery occlusion is described. From the multiexponential relaxation, the presence of two T(2) relaxation times in the range of 0.03-0.5 s, T(2A) (shortest) and T(2B) (longest), with very different relative weights (respectively, A and B), is evidenced. In our models of cerebral damage, the changes in A and B were more evident than those in T(2A) and T(2B). The two T(2) values were interpreted as belonging to water molecules in two different compartments; therefore, the difference between the damaged and normal regions revealed by means of standard T(2)-weighted images is suggested to be due to a different water distribution in the two compartments, rather than different T(2)'s. The T(2) relaxation in the SHR-SP stroke model is analyzed for the first time using a multiexponential method. The power of a detailed analysis of MRI relaxation times is confirmed by the correspondence between the revealed changes in T(2A), T(2B), A and B, and the known T(2)W and DWI results about blood-brain barrier functionality.

Unfavorable effect of photodynamic therapy for late subretinal neovascularization with chorioretinal anastomoses associatedwith idiopathic multiple serous detachments of the retinal pigment epithelium.

PURPOSE: To describe a case of a 46-year-old woman with an asymptomatic history of unilateral multiple serous detachments of the retinal pigment epithelium (PED) in the right eye, treated with photodynamic therapy (PDT) with verteporfin for recent onset of subfoveal choroidal neovascularization (CNV) with chorioretinal anastomoses (CRA). RESULTS: Fluorescein and indocyanine green (ICG) angiography, performed with a Heidelberg scanning laser ophthalmoscope (SLO), demonstrated a predominantly classic foveal choroidal neovascular membrane associated with a PED and 1 one retinal and 2 two venous chorioretinal anastomoses. The left fundus was normal. PDT therapy was performed according to standard techniques. Three PDT treatments were performed at an interval of 3 months. Three months after the second PDT, visual acuity dropped to 20/200, with an en-largement of the neovascular network. One month after the third treatment, visual acuity deteriorated further and the CRA appeared enlarged, associated with a dense fibrotic re-action in the centere of the lesion. CONCLUSIONS: This clinical observation demonstrates that idiopathic serous detachments of the retinal pigment epithelium may represent predisposing changes to CNV development, and in the case CNV is associated with CRA, PDT may be unsuccessful. (Eur J Ophthalmol 2004; 14: 568-71).

Unfavorable effect of photodynamic therapy for late subretinal neovascularization with chorioretinal anastomoses associated with idiopathic multiple serous detachments of the retinal pigment epithelium.

PURPOSE: To describe a case of a 46-year-old woman with an asymptomatic history of unilateral multiple serous detachments of the retinal pigment epithelium (PED) in the right eye, treated with photodynamic therapy (PDT) with verteporfin for recent onset of subfoveal choroidal neovascularization (CNV) with chorioretinal anastomoses (CRA). METHODS: Case report. RESULTS: Fluorescein and indocyanine green (ICG) angiography, performed with a Heidelberg scanning laser ophthalmoscope (SLO), demonstrated a predominantly classic foveal choroidal neovascular membrane associated with a PED and 1 one retinal and 2 two venous chorioretinal anastomoses. The left fundus was normal. PDT therapy was performed according to standard techniques. Three PDT treatments were performed at an interval of 3 months. Three months after the second PDT, visual acuity dropped to 20/200, with an enlargement of the neovascular network. One month after the third treatment, visual acuity deteriorated further and the CRA appeared enlarged, associated with a dense fibrotic reaction in the centere of the lesion. CONCLUSIONS: This clinical observation demonstrates that idiopathic serous detachments of the retinal pigment epithelium may represent predisposing changes to CNV development, and in the case CNV is associated with CRA, PDT may be unsuccessful.

Immunohistochemical study of human pterygium.

The purpose of this study has been to evaluate the immunohistochemical characteristics of human pterygial tissues in order to ascertain the possible contribution of an immunological mechanism in the pathogenesis of pterygium and to investigate the presence in the pterygial tissues of some melanoma-associated antigens, in order to evaluate if there may be a small possibility of correlation of the two diseases. Human biopsy specimens of pterygium were obtained by surgery for pterygium excision. Tissue segments were fixed and processed for paraffin embedding. Microtome sections were treated for the immunohistochemical demonstration of IgA, IgM, IgG, CD3, CD20, CD68, HLA-DR, Protein S100, HMB45, and Melan A using the avidin-biotin peroxidase method or the streptavidin biotin-alkaline phosphatase method. The findings suggest that all the effector components of the mucosal immune system are present in the human pterygium and, among the most sensitive markers for melanoma, only S100 shows immunoreactivity. An immunopathogenetic mechanism seems to be responsible for the pathogenesis of pterygium, perhaps being caused by pre-existing conjunctivitis or microtrauma in combination with the patient's predisposition. No correlation between pterygium and melanoma was found.

The presence of a local immune system in the upper blind and lower part of the human nasolacrimal duct.

The nasolacrimal duct is exposed to exogenous agents, including potentially harmful microorganisms, coming from the eye surface by the lacrimal sac, and from the nasal cavity by the inferior meatus of the nose. The upper blind and lower part of the human nasolacrimal duct were examined immunohistochemically to ascertain the presence and localization of immunoglobulin-producing cells and the epithelial expression of IgA, IgM, and IgG in order to verify the possible antimicrobial properties of this duct. IgA-, IgM-, and IgG-positive immunocompetent cells were recognizable in the lamina propria of the upper blind and lower part of the human nasolacrimal duct, while an evident immunoreactivity for sIgA, IgM, and IgG was demonstrated in the cytoplasm of the apical epithelial cells. The results suggest that all the effector components of the mucosal immune system are present in that area of the human nasal mucosa next to the opening of the nasolacrimal duct as well as in the human lacrimal sac.

Blue-yellow perimetry in patients with ocular hypertone.

The authors report the data of the blue-yellow (B-Y) perimetry compared with the Standard perimetry in normal subjects with endocular hypertension or with initial glaucoma. With the aim of evaluating the relationship with chromatic sense deficits, precociously found in glaucoma, the F-M 100 Hue test and Lanthony D 15 Desaturé test were done. Checks were made of refraction, visual acuity, pupil diameter and assumption of medications. Sensitivity reduction in eyes with initial glaucoma is highly significant with the B-Y perimetry. Pupil diameter reduction is quite uninfluential while the chromatic sense shows some quantitative and qualitative deficits.

The influence of diabetes mellitus on primary open angle glaucoma perimetry.

We have carried out a study into retinal sensitivity alterations in the course of primary open angle glaucoma to see if their appearance and evolution might be influenced by concomitant diabetes mellitus. The visual field examination (Perimeter Octopus 500 EZ, programme G1) indicated prevalent sensitivity defects in the superior hemifield, both in glaucoma only subjects and in those with diabetes as well. As to the inferior hemifield, a greater, statistically significant, retinal sensitivity defect was found in the inferior temporal quadrant of the left eye in the group of diabetics.

Genetic mapping of autosomal dominant primary open-angle glaucoma (POAG) in Sardinia.

Primary open-angle glaucoma (POAG) can be subdivided into two groups according to age of onset: (1) the more prevalent middle to late-age-onset chronic open-angle glaucoma (COAG) diagnosed after age 40, and (2) the less common form, juvenile open-angle glaucoma (JOAG), which occurs between 3 years of age and early adulthood. Susceptibility to either COAG or JOAG has been found to be inherited. The discovery of several genetic markers spanning the region 1q21-q24 in genetic linkage with autosomal dominant juvenile open-angle glaucoma (adJOAG) represents a major breakthrough towards the localisation of gene(s) responsible for the disease. Linkage analysis is a powerful means of distinguishing disease loci in large families with dominant disease. However the size of the group of families may represent a crucial factor for the linkage analysis. Sardinia is an island with a relatively isolated ethnic group showing a relatively high frequency of ad JOAG and COAG (Fossarello et al, 1994) and it is genetically more homogeneous than most Western populations. Therefore it represents an ideal ethnic group to search for linkage. We identified 18 families affected by POAG in which the disease appears to be inherited as autosomic dominant trait. In all families but two, occurrence of both JOAG and COAG in the same kindred was observed. Identification of adPOAG locus was performed by linkage analysis using 9 microsatellite markers spanning the region 1q21-q24. No significant linkage was observed. Our findings provide further evidence for genetic heterogeneity in autosomal dominant primary open angle glaucoma, even in a geographic area where a relatively homogeneous genetic background exists.

Evaluation of efficacy and tolerability of Nilutamide and Buserelin in the treatment of advanced prostate cancer.

Several different medical strategies have been proposed for the treatment of advanced prostatic cancer: androgen withdrawal by surgical castration on indirect suppression of androgen production by estrogen or estrogen-like substances, antiandrogen compounds or LH-RH analogues. The Authors evaluated in detail tolerability and efficacy of a combination therapy of a LHRH analogue (Buserelin) and a pure antiandrogen (Nilutamide) in a group of 15 patients with advanced prostate cancer (stage D) followed over a period of six months.