Quantitative analysis of optic nerve damage in idiopathic intracranial hypertension (IIH) at diagnosis.

Optic neuropathy secondary to idiopathic intracranial hypertension (IIH) may be a severe complication which must be early identified, adequately monitored and treated to avoid blindness. The aim of this study was to quantify optic nerve involvement at time of diagnosis in a prospectively series of IIH investigated at a single Institution and to identify objective parameters for early diagnosis and follow-up. 38 consecutive patients (9 men, 29 females, mean age 39.8 years) with IIH underwent a complete neuro-ophthalmological evaluation including standardized automated perimetry as functional measurement of optic neuropathy and spectral domain optical coherence tomography (SD-OCT) measurements to grade papilledema or optic nerve atrophy. An overall diagnosis of optic nerve involvement was made in 50 out of 76 eyes (66 %); ophthalmoscopic signs of papilledema were identified in 35 eyes (46 %) while optic disc pallor was found in 13 (17 %). In all patients mean visual field deviation (MD, dB) was -7.2 (range 5.3-33.2). SD-OCT measurements of peripapillary retinal nerve fiber layer thickness (PRNFLT) and of macular ganglion cell complex thickness (MGCCT) obtained in 40 eyes (20 subjects) showed normal PRNFLT in 12 eyes (30 %), increased in 16 (40 %) and reduced in 12 eyes (30 %); normal MGCCT in 26 eyes (65 %), reduced in 14 (35 %). In all eyes average RNFLT was increased (mean 130 µm, range 219-59) and average MGCCT was decreased compared to normal values (mean 89.5 µm, range 198-65). Increased PRNFLT was associated with reduced MGCCT in 4 eyes (10 %) indicating early retrograde optic nerve damage. Decreased PRNFLT was associated with decreased MGCCT in 10 eyes (83 %). These results indicate that, in IIH patients, signs of optic neuropathy can be identified in more than half of cases, even without papilledema evidenced on ophthalmoscopic examination. Moreover, an SD-OCT analysis, which can be definitively useful to quantify optic nerve edema or atrophy, can show damage of retinal ganglion cells in an early phase of the disease.

Headache prevalence and clinical features in patients with idiopathic intracranial hypertension (IIH).

Headache is a key symptom of idiopathic intracranial hypertension (IIH). Operational diagnostic criteria for "Headache attributed to IIH" are included in the international classification of headache disorders, the ICHD-2. The association of IIH with obesity was established by several reports. We investigate the prevalence of headache and its main clinical features in a clinical sample of IIH patients. The possible correlations between the presence of headache and body mass index (BMI) and intracranial pressure (ICP) levels were studied in a consecutive clinical series of patients, in whom diagnosis of IIH was confirmed by exclusion of secondary forms and by the evidence of increased ICP. Differences for age, BMI, and ICP between patients with and without headache and between males and females were assessed with Mann-Whitney U test. Spearman's correlation analysis was used to assess relationships between age, BMI, and ICP. P value < 0.05 was used to set statistical significance. 40 patients entered the study (9 males, 31 females; mean age 39, 8 years, SD 13.2). Headache was reported by 75 % patients. Those characteristics which are included in the present international diagnostic criteria for "Headache attributed to IIH" were reported by a remarkable proportion of the studied patients, but not by all. On the other hand, some headache features usually attributed to migraine forms, and which are not among the required criteria were present in some patients: pulsating quality and unilateral distribution of pain in around 20 %, and migrainous associated symptoms in more than 40 % of the sample. According to statistical analyses, no differences were found for age, BMI, and ICP between patients with and without headache. Our results confirmed the strong association between headache and IIH. Although no significant correlations between some of the key features of IIH were found in this study, we suggest that further studies on larger series--possibly with a longitudinal evaluation--are needed, to help clinicians in categorizing different subgroups among IIH patients as well as in identifying the main factors influencing the prognosis of this disorder.

Immunohistochemical detection of the p27 capsid protein of caprine arthritis-encephalitis virus (CAEV) in bone-marrow cells of seropositive goats.

Bone-marrow samples were collected from 48 CAEV-seropositive, symptomless goats (30 kids, 18 adults). The samples were formalin-fixed and processed for histological examination. In addition, all samples were examined immunohistochemically with a monoclonal antibody (1A7) against the p27 capsid protein of maedi-visna virus, an antibody which cross-reacts with the Ca-p27 of CAEV. Samples from 16 goats (10/30 kids, 6/18 adults) showed positive immunolabelling of bone-marrow stromal cells (fibrocytes, endothelial cells and adipocytes) and of scattered macrophages, whereas haematopoietic cells were negative. The detection of viral Ca-p27 protein in bone-marrow fibrocytes was consistent with previous in-vitro studies which indicated that such cells are semi-permissive for CAEV infection. It is speculated that bone-marrow stromal cells represent a viral reservoir in symptomless animals.

Fusion of the distal sesamoid and distal phalanx in a yearling colt.

This paper describes a rare malformation of the distal portion of the left foredigit of a 15-month-old half-bred colt which was severely lame. Radiological, pathological and tomographic studies revealed hypoplasia of the metacarpophalangeal skeleton, absence of the distal sesamoid and deformation of the distal phalanx. Morphological analysis of the lesion and computerised measurement of the width of the articular surface of the distal phalanx suggested an early fusion between the distal sesamoid and distal phalanx.

Conserved structure and promoter sequence similarity in the mouse and human genes encoding the zinc finger factor BERF-1/BFCOL1/ZBP-89.

We have characterized the genomic structure of the mouse Zfp148 gene encoding Beta-Enolase Repressor Factor-1 (BERF-1), a Kruppel-like zinc finger protein involved in the transcriptional regulation of several genes, which is also termed ZBP-89, BFCOL1. The cloned Zfp148 gene spans 110 kb of genomic DNA encompassing the 5'-end region, 9 exons, 8 introns, and the 3'-untranslated region. The promoter region displays the typical features of a housekeeping gene: a high G+C content and the absence of canonical TATA and CAAT boxes consistent with the multiple transcription initiation sites determined by primary extension analysis. Computer-assisted search in the human genome database allowed us to determine that the same genomic structure with identical intron-exon organization is conserved in the human homologue ZNF 148. Functional analysis of the 5'-flanking sequence of the mouse gene indicated that the region from nucleotide -205 to +144, relative to the major transcription start site, contains cis-regulatory elements that promote basal expression. Such sequences and the overall promoter architecture are highly conserved in the human gene. Furthermore, we show that the complex transcription pattern of the Zfp148 gene might be due to a combination of alternative splicing and differential polyadenylation sites utilization.

Using a panel of monoclonal antibodies to detect Maedi virus (MV) in chronic pulmonary distress of sheep.

A selected panel of six monoclonal antibodies (mAbs) against Maedi-Visna virus (MVV), recognising the core proteins (p27 and p15) and the envelope protein (gp105) of MVV, was tested using different unmasking techniques on paraffin embedded lung samples of a seropositive sheep. Only three mAbs were chosen, according to their strong reactivity. mAbs 1A7, 1B6 and 4B3 were employed in an immunohistochemical trial focused on the diagnosis of the lungs of 26 sheep with progressive pulmonary distress. These mAbs demonstrated MVV in 21 out of 26 cases including lymphoid interstitial pneumonia (LIP) and pulmonary adenomatosis. In only nine cases did all three mAbs react positively with the same sample. The sensitivity of immunohistochemical diagnosis of Maedi pneumonia can be increased by using mAbs 1A7, 4B3 and 1B6 together; that is a panel of mAbs direct against the envelope (gp105) and capsid (p27) viral proteins. The positive signal was focal and confined to the cytoplasm of bronchoalveolar epithelial cells and alveolar-interstitial macrophages. The results suggest that this panel of mAbs is useful to confirm severe LIP lesions such as Maedi pneumonia, to demonstrate Maedi infections in mild LIP, to demonstrate MVV in mixed pulmonary changes, and to investigate the pathogenesis of Maedi-Visna.

Phosphorylation-dependent regulation of skeletogenesis in sea urchin micromere-derived cells and embryos.

Sea urchin embryo micromeres when isolated and cultured in vitro differentiate to produce spicules. Although several authors have used this model, almost nothing is known about the signaling pathways responsible for initiating skeletogenesis. In order to investigate the potential involvement of phosphorylation events in spiculogenesis, the effect of inhibitors of protein kinases and phosphatases on skeleton formation was studied. Results obtained using both cultured micromeres and embryos revealed that protein tyrosine kinase and phosphatase inhibitors blocked skeleton formation, but not serine/threonine phosphatase inhibitors. The inhibitors showed a dose-dependent effect and when removed from micromere or embryo culture, spicule formation resumed. Inhibition of tyrosine phosphatases resulted in an increase in the tyrosine phosphorylation level of two major proteins and a modest decrease in the expression of the mRNA coding for type I fibrillar collagen. These findings strongly suggest that tyrosine phosphorylation and dephosphorylation is required for micromere differentiation and for normal skeletogenesis during sea urchin embryo development.

Some aspects of humoral and cellular immunity in naturally occuring feline infectious peritonitis.

Haematology, antibody titers and serum protein electrophoresis from 48 cats (34 effusive and 14 noneffusive forms) affected with feline infectious peritonitis (FIP) were studied and compared with those of 20 healthy cats. In the effusive form, antibody titers and protein electrophoresis in the effusions were analyzed. The distribution of the immune cells and of the virus in FIP lesions were also investigated immunohistochemically with the avidin-biotin complex (ABC) method, using antibodies against the FIP virus (FIPV), myelomonocytic (MAC387) and lymphoid (CD3, CD4 and CD8 for T-cells and IgM and IgG for B-cells) antigens. Seropositive animals (antibody titer>1:100) were present among both the FIP infected cats (73%) and the healthy cats (70%). Cats with effusive FIP had neutrophilic leukocytosis (P>0.05), lymphopenia (P<0.01) and eosinopenia (P<0.001). In both effusive and noneffusive forms decreased albumin/globulin ratio (P<0.001) with hypoalbuminemia (P<0.001), hyperglobulinemia (P<0.001) and increased alpha2- (P<0.05), beta- (P<0.05) and gamma-globulins (P<0.001) were found. Hypergammaglobulinemia was not related to the antibody titers, suggesting the presence of other proteins with gamma-motility (e.g. complement fractions). The electrophoretic pattern of the effusions was always similar to that of the corresponding serum. Antibody titers higher than those of the corresponding serum were often detected in the effusions. Immunohistochemical findings were not related to the antibody titers, but they were related to the histological aspect of the lesions. In cellular foci of FIP lesions many virus-infected macrophages and few lymphocytes, mainly CD4+, were found. Extracellular viral and myelomonocytic antigens were also detectable in the foci with intercellular necrosis. Only few FIPV-infected cells were present at the periphery of the larger necrotic foci: in these lesions MAC387+ cells were mainly neutrophils, with many MAC387 macrophages, probably due to their activated state; a small number of lymphocytes, with an increasing percentage of CD8+ cells was present. Lymphocytes were more abundant when cellular foci and FIP-infected macrophages were centered around neoformed vessels. IgM and IgG exposing B-cells were always few and scattered. In conclusion the simultaneous analysis of body fluids and of the cellular composition of the lesions showed a complex immune status, on which type III and type IV hypersensitivity could coexist.

Tetracycline inhibits the nitric oxide synthase activity induced by endotoxin in cultured murine macrophages.

Here we investigate the effects of tetracycline base and of a semi-synthetic tetracycline derivative, doxycycline, on the induction of inducible nitric oxide synthase and, hence, on the production of nitric oxide (NO) by lipopolysaccharide in J774 macrophage cultured in vitro. The treatment of J774 line with tetracycline base (6.25-250 microM) or doxycycline (5-50 microM) dose-dependently decreased the lipopolysaccharide-stimulated (1 microg/ml) inducible NO synthase activity and, consequently, nitrite formation. For instance, the inhibition was 70% for tetracycline base at 250 microM and 68% for doxycycline at 50 microM. The inhibitory effect of tetracyclines was due neither to a reduction in the viability of the cells, studied as colorimetric 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromide (MTT) reduction assay, nor to an indiscriminate inhibition of total protein synthesis, but to a specific decrease in inducible NO synthase protein content in the cells, as attested by the significant reduction of the expression of inducible NO synthase, assayed by sodium-dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. However, no effect of tetracyclines on inducible NO synthase mRNA accumulation could be demonstrated in lipopolysaccharide-stimulated macrophage line, suggesting that the inhibitory effect of tetracyclines on NO synthesis involves post-transcriptional events. The reduction in lipopolysaccharide-stimulated nitrite accumulation produced by tetracyclines was significantly less when they were applied 6 h after lipopolysaccharide and absent 12 h after lipopolysaccharide, indicating that tetracyclines modify an early event in inducible NO synthase activation operating after mRNA transcription. The findings presented in this study indicate that the modulation of NO synthesis is another possible pathway by which tetracyclines may function as anti-inflammatory compounds.

Type IV hypersensitivity in the pathogenesis of FIPV-induced lesions.

In focal lesions of feline infectious peritonitis (FIP), the cells involved in the delayed-type hypersensitivity were identified in formalin-fixed paraffin-embedded and frozen samples taken from 35 affected cats. The clinical diagnosis of FIP was confirmed by necropsy, histology and direct immunofluorescence against the coronaviruses on cryostatic sections. The immune cells were detected immunohistochemically by the Avidin-Biotin-Complex (ABC) method using either polyclonal antibodies against lymphoid antigens (CD3) or monoclonal antibodies against lymphoid (PAN-T, CD4, CD8) and myeloid antigens (MAC387). Better identification of T cells and macrophages was found on formalin-fixed paraffin-embedded sections than on cryostatic ones, while T lymphocyte subpopulations could be differentiated only in cryostatic sections. Type IV hypersensitivity was detected in focal feline infectious peritonitis virus (FIPV)-induced lesions from progressive activation of T lymphocytes, mainly CD4+, and the presence of granulocytes and macrophages. The FIPV-induced lesions could be studied as examples of granulomas caused by unconventional antigens, such as viruses or immune complexes.

Modulation of nitric oxide production by tetracyclines and chemically modified tetracyclines.

Chemically modified tetracyclines (CMTs) dose-dependently decreased inducible nitric oxide synthase (iNOS) and, consequently, nitric oxide (NO) formation by the lipopolysaccharide (LPS)-stimulated J774 line. The inhibitory effect was due to a specific reduction in the iNOS protein content in the cells, as attested by Western blot analysis and by the inhibition of iNOS mRNA accumulation. Furthermore, CMTs cause a dose-dependent increase in cell death in the J774 line mediated by the NO-independent apoptotic mechanism.

The gene encoding the transcriptional repressor BERF-1 maps to a region of conserved synteny on mouse chromosome 16 and human chromosome 3 and a related pseudogene maps to mouse chromosome 8.

We have recently identified and characterized a Kruppel-like zinc finger protein (BERF-1), that functions as a repressor of beta enolase gene transcription. By interspecific backcross analysis the gene encoding BERF-1 was localized 4.7 cM proximal to the Mtv6 locus on mouse chromosome 16, and an isolated pseudogene was localized to mouse chromosome 8, about 5.3 cM distal to the D8Mit4 marker. Nucleotide sequence identity and chomosome location indicate that the gene encoding BERF-1 is the mouse homologue (Zfp148) of ZNF148 localized to human chromosome 3q21, a common translocation site in acute myeloid leukemia patients.

[Pulmonary changes in the early phase of contagious bovine pleuropneumonia. Histologic and immunohistochemical aspects]. / Alterazioni polmonari della Pleuropolmonite Contagiosa Bovina (PPCB) in fase esordiente. Aspetti istologici ed immunoistochimici.

Recent pulmonary lesions at slaughter in bovines affected by Mycoplasma mycoides during the years 1990-1993 in Northern Italy are reported. Several histologic and immunocytochemical examinations showed that: the earliest lesions of CBPP are foci of desquamative bronchiolo-alveolitis, the highest concentration of the antigen is found in the cytoplasm of desquamated alveolar macrophages. From our results, we can exclude that CBPP begins with interstitial lesions as some authors have hypothesized. On the contrary, the hypothesis, formulated at the beginning of the century, that CBPP begins with a desquamative bronchiolo-alveolitis seems to be more convincing. Our results give some help to construct a correct pathogenesis of the disease.

HLA-B8,DR3 phenotype and lymphocyte responses to phytohaemagglutinin.

Several reports have shown that HLA-B8,DR3 positive subjects may display some changes in immune parameters when compared with HLA-B8,DR3 negative ones and are prone to develop several immunological diseases. In the present study we have analysed the proliferative response to phytohaemagglutin (PHA) in HLA-typed healthy subjects. A twin method was also employed to assess the role of genetic and environmental factors in the regulation of the response to the mitogen. It was not possible to demonstrate any difference in proliferative response to optimal doses of PHA between groups of subjects carrying or not carrying the HLA-B8,DR3 phenotype. When suboptimal responses were studied, however, the results showed that lymphocyte responses were significantly decreased in HLA-B8,DR3 positive subjects compared with the negative ones. Moreover, the experiments performed with twins demonstrated that environmental factors were more important than genetic factors in the proliferative response to mitogen. The fact that the HLA-B8,DR3 phenotype affects the suboptimal response to PHA although environmental factors are more important than genetic factors in the response to the mitogen seems of some interest. However, these results could be consistent with the high incidence of autoimmune disorders among HLA-B8,DR3 positive individuals.

HLA-Cw4 association with acute lymphoblastic leukaemia in Sicilian patients.

The HLA frequencies of 50 Sicilian patients affected with acute lymphoblastic leukaemia (ALL) were examined. The frequency of Cw4 antigen was significantly increased in patients. Thus results obtained in our homogeneous population confirm in part previous reports suggesting that Cw4-related genetic factors might be involved in the susceptibility to aetiological or pathogenetic mechanisms which play a role in some haematological malignancies.

Histopathologic observations on Sertoli cell tumors in dog.

Thirty-three cases of Sertoli cell tumor, occurring in dogs of different breeds and ages, were studied histologically. Ectopic testes showed a particularly high tendency to develop the tumor. Metastases were not detected in regularly autopsied dogs nor reported or later ascertained for the bioptic consignments. According to Nielsen and Lein's classification (1974) our findings were divided as follows: 27 intratubular Sertoli cell tumors, 19 with stromal invasion and 8 without invasion; 2 diffuse tumors; 4 multiple primary tumors (3 Sertoli-seminoma cell tumor and 1 Sertoli-Leydig cell tumor). The above classification is discussed and proposed tentatively for revision.