Plasma total prion protein as a potential biomarker for neurodegenerative dementia: diagnostic accuracy in the spectrum of prion diseases.

AIMS: In the search for blood-based biomarkers of neurodegenerative diseases, we characterized the concentration of total prion protein (t-PrP) in the plasma of neurodegenerative dementias. We aimed to assess its accuracy in this differential diagnostic context. METHODS: Plasma t-PrP was measured in 520 individuals including healthy controls (HC) and patients diagnosed with neurological disease control (ND), Alzheimer's disease (AD), sporadic Creutzfeldt-Jakob disease (sCJD), frontotemporal dementia (FTD), Lewy body dementia (LBD) and vascular dementia (VaD). Additionally, t-PrP was quantified in genetic prion diseases and iatrogenic CJD. The accuracy of t-PrP discriminating the diagnostic groups was evaluated and correlated with demographic, genetic and clinical data in prion diseases. Markers of blood-brain barrier impairment were investigated in sCJD brains. RESULTS: Compared to HC and ND, elevated plasma t-PrP concentrations were detected in sCJD, followed by FTD, AD, VaD and LBD. In sCJD, t-PrP was associated neither with age nor sex, but with codon 129 PRNP genotype. Plasma t-PrP concentrations correlated with cerebrospinal fluid (CSF) markers of neuro-axonal damage, but not with CSF t-PrP. In genetic prion diseases, plasma t-PrP was elevated in all type of mutations investigated. In sCJD brain tissue, extravasation of immunoglobulin G and the presence of swollen astrocytic end-feet around the vessels suggested leakage of blood-brain barrier as a potential source of increased plasma t-PrP. CONCLUSIONS: Plasma t-PrP is elevated in prion diseases regardless of aetiology. This pilot study opens the possibility to consider plasma t-PrP as a promising blood-based biomarker in the diagnostic of prion disease.

Magnetic resonance spectroscopy in intracranial tumours of glial origin.

BACKGROUND AND PURPOSE: To determine in vivo magnetic resonance spectroscopy (MRS) characteristics of intracranial glial tumours and to assess MRS reliability in glioma grading and discrimination between different histopathological types of tumours. MATERIAL AND METHODS: Analysis of spectra of 26 patients with glioblastomas, 6 with fibrillary astrocytomas, 4 with anaplastic astrocytomas, 2 with pilocytic astrocytoma, 3 with oligodendrogliomas, 3 with anaplastic oligodendrogliomas and 17 control spectra taken from healthy hemispheres. RESULTS: All tumours' metabolite ratios, except for Cho/Cr in fibrillary astrocytomas (p = 0.06), were statistically significantly different from the control. The tumours showed decreased Naa and Cr contents and a high Cho signal. The Lac-Lip signal was high in grade III astrocytomas and glioblastomas. Reports that Cho/Cr ratio increases with glioma's grade whereas Naa/Cr decreases were not confirmed. Anaplastic astrocytomas compared to grade II astrocytomas had a statistically significantly greater mI/Cr ratio (p = 0.02). In pilocytic astrocytomas the Naa/Cr value (2.58 ± 0.39) was greater, whilst the Cho/Naa ratio was lower (2.14 ± 0.64) than in the other astrocytomas. The specific feature of oligodendrogliomas was the presence of glutamate/glutamine peak Glx. However, this peak was absent in two out of three anaplastic oligodendrogliomas. Characteristically, the latter tumours had a high Lac-Lip signal. CONCLUSIONS: MRS in vivo cannot be used as a reliable method for glioma grading. The method is useful in discrimination between WHO grade I and WHO grade II astrocytomas as well as oligodendrogliomas from other gliomas.

Differential diagnosis of intracranial meningiomas based on magnetic resonance spectroscopy.

BACKGROUND AND PURPOSE: To determine in vivo magnetic resonance spectroscopy (MRS) characteristics of intracranial meningiomas and to assess MRS reliability in meningioma grading and discrimination from tumours of similar radiological appearance, such as lymphomas, schwannomas and haemangiopericytomas. MATERIAL AND METHODS: Analysis of spectra of 14 patients with meningiomas, 6 with schwannomas, 2 with lymphomas, 2 with haemangiopericytomas and 17 control spectra taken from healthy hemispheres. RESULTS: All the patients with meningiomas had a high Cho signal (long TE). There were very low signals of Naa and Cr in the spectra of 10 patients. A reversed Ala doublet was seen only in 2 cases. Four patients had a negative Lac signal, whereas 3 had high Lac-Lip spectra. Twelve spectra showed high Cho signals (short TE). In one case the Cho signal was extremely low. All spectra displayed a very low Cr signal, but high Glx and Lac-Lip signals. Ala presence was found only in 3 patients. The mean Cho/Cr ratio (PRESS) was 5.97 (1.12 in normal brain, p < 0.05). Lac-Lip was present in all the meningiomas (STEAM). The Ala signal was seen only in 2 spectra with long TE and in 3 sequences of the short TE sequences. There were both ß/γ-Glx and α-Glx/glutathione signals in all 14 meningiomas. CONCLUSIONS: MRS is unable to discriminate low and high grade meningiomas. The method seems to be helpful in discriminating lymphomas (absent Glx signal), schwannomas (mI signal in the short TE sequences) and haemangiopericytomas (presence of mI band) from meningiomas.

Ultrastructural study of florid plaques in variant Creutzfeldt-Jakob disease: a comparison with amyloid plaques in kuru, sporadic Creutzfeldt-Jakob disease and Gerstmann-Sträussler-Scheinker disease.

BACKGROUND: Although the histological features of the amyloid plaques in variant Creutzfeldt-Jakob disease (vCJD) are distinct from those in other forms of prion disease [kuru, sporadic Creutzfeldt-Jakob disease (sCJD) and Gerstmann-Sträussler-Scheinker disease (GSS)], their ultrastructural features have only been described in a single case report. AIMS: To study vCJD plaques systematically and compare them with plaques in kuru, sCJD, GSS and Alzheimer disease (AD). METHODS: Amyloid plaques were studied by transmission electron microscopy and image analysis in five cases of vCJD, three cases of GSS, two cases of sCJD, one case of kuru and five cases of AD. Immunohistochemistry was performed on paraffin sections from one case of vCJD, two cases of GSS, one case of kuru and two cases of sCJD. RESULTS: The florid plaques in vCJD were either compact or more diffuse; in both forms, the radiating fibrils were organized into thick 'tongues', in contrast to kuru plaques. Dystrophic neurites (DNs) containing lysosomal electron-dense bodies or vesicles surrounded florid plaques. Microglial cells were found within florid plaques; occasional amyloid fibrils were identified in membrane-bound pockets of microglial cells. In vCJD, there was significant tau immunoreactivity in DNs around florid plaques while, in sCJD, GSS and kuru, minimal tau immunoreactivity was observed around plaques. CONCLUSIONS: The ultrastructure of the florid plaques and DNs in vCJD is more reminiscent of neuritic plaques in AD than kuru or multicentric plaques. These findings may reflect differences both in the strains of the transmissible agents responsible for these disorders and in host factors.

BCR expression is decreased in meningiomas showing loss of heterozygosity of 22q within a new minimal deletion region.

Neurofibromin 2 (NF2), located on chromosome arm 22q, has been established as a tumor suppressor gene involved in meningioma pathogenesis. In our study, we investigated 149 meningiomas to determine whether there are additional tumor suppressor genes localized on chromosome 22q, apart from NF2, that might be involved in meningioma pathogenesis. The LOH analysis on chromosome 22q identified two regions of deletion: the first one, which is limited to the NF2 gene locus, and the second one, which is outside this location. The new minimal deletion region (MDR) included the following genes: BCR (breakpoint cluster region), RAB36 (a member of RAS oncogene family), GNAZ [guanine nucleotide binding protein (G protein), alpha-z polypeptide], and RTDR1 (rhabdoid tumor deletion region gene 1). The expression levels of all these genes, including NF2, were subsequently analyzed by quantitative real-time polymerase chain reaction. We observed a significantly lowered expression level of NF2 in meningiomas with 22q loss of heterozygosity (LOH) within NF2 region compared to the one in meningiomas with 22q retention of heterozygosity (ROH, P<0.05). Similarly, BCR showed a significantly lowered expression in meningiomas with 22q LOH within the new MDR compared to cases with 22q ROH (P<0.05). Our data, together with the already published information considering BCR function suggest that BCR can be considered as a candidate tumor suppressor gene localized on chromosome 22q which may be involved in meningioma pathogenesis.

Molecular analysis of chromosome 1, 10 and 19 abnormalities in human oligodendroglial tumors: relationship between frequency of LOH grade, age and gender.

BACKGROUND: Loss of heterozygosity (LOH) on 1p and 19q is observed in most oligodendroglial tumors. LOH on 10q appears to be less common in these tumors as compared to other gliomas. PATIENTS AND METHODS: We reviewed 14 patients with oligodendroglial tumors (10 low-grade and 4 anaplastic oligodendroglioma) to evaluate the frequency of LOH on 1p, 10q and 19q and correlate it with tumor grade and patients' age and gender; 5 loci on 1p and 5 on 19q as well as 4 on 10q were analyzed for LOH using PCR techniques. RESULTS: LOH on 1p together with 19q was detected in 6 tumors, 1 tumor showed deletion of 19q accompanied with deletion on 10q. Deletion on 1p was associated with deletion of 19q (p < 0.005) and mutual associations among deletions at loci on 19q (p < 0.05) were found. Patients with LOH on 1p were younger on average than patients with retained heterozygosity (p = 0.05). Grade II oligodendrogliomas predominated among younger patients (p < 0.01) while grade III oligodendrogliomas predominated among women (p < 0.005). No association between LOH on 1p nor 19q and tumor grade or patients' gender was found. CONCLUSION: Our study provides several clinically interesting findings and further supports the hypothesis of chromosome 1p and 19q involvement in the oligodendroglial cancerogenesis.

GADD45A and EPB41 as tumor suppressor genes in meningioma pathogenesis.

Deletions of 1p occur in approximately 30% of meningiomas. Based on loss of heterozygosity (LOH) analysis, two regions on 1p have been suspected to be carriers of tumor suppressor genes. We chose the GADD45A and EPB41 genes as tumor suppressor candidates based on their function and chromosomal localization. We analyzed 19 cases of meningioma with LOH of 1p by means of sequencing of the GADD45A gene and Western blotting of the GADD45a protein. Twenty cases of meningioma without 1p LOH were also analyzed by Western blotting to find out if changes of the GADD45a protein expression occurred. Nineteen samples with 1p LOH (12 grade I; 7 grade II, WHO classification) and 20 samples without 1p LOH (18 grade I; 2 grade II) were also analyzed by means of real-time polymerase chain reaction to find abnormalities in EPB41 mRNA levels in meningioma. LOH analysis was performed using seven microsatellite markers: D1S508 (1p36.2), D1S199 (1p36.1) D1S2734 (1p36.1), D1S2720 (1p34), D1S197 (1p32), D1S162 (1p32), D1S429 (1p11). LOH analysis confirmed previously described localization of putative tumor suppressor genes on 1p and involvement in meningioma pathogenesis (1p36 and 1p32). The open reading frame of GADD45A and intron splicing sites showed neither mutations nor polymorphisms. GADD45a protein molecular weight and expression level were unaltered in meningiomas with and without 1p LOH. We conclude that the GADD45A gene is not involved in meningioma tumorigenesis. EPB41 gene expression was unchanged in all analyzed meningiomas. This suggests that involvement of the EPB41 gene (4.1R protein) in meningioma pathogenesis should be reconsidered.

Ultrastructural pathology of prion diseases revisited: brain biopsy studies.

We report here a detailed ultrastructural comparison of brain biopsies from 13 cases of Creutzfeldt-Jakob disease (CJD) and from one case of fatal familial insomnia (FFI). The latter disease has not heretofore benefited from ultrastructural study. In particular, we searched for tubulovesicular structures (TVS), 35-nm particles regarded as the only disease-specific structures at the level of thin-section electron microscopy. Our material consisted of brain biopsies obtained by open surgery from one FFI case from a new French family, one case of variant CJD (vCJD), nine cases of sporadic CJD (sCJD), two cases of iatrogenic (human growth hormone) CJD and one case of hereditary CJD (Val203Iso). The ultrastructural picture of the cerebral cortex of the FFI patient was virtually indistinguishable from that of CJD. TVS were found, albeit only after prolonged search. Typical spongiform change was observed, consisting of intracellular membrane-bound vacuoles containing secondary chambers (vacuoles within vacuoles) and amorphous material. Neuronal degeneration was widespread: some processes contained degenerating mitochondria and lysosomal electron-dense bodies and these met the criteria for neuroaxonal dystrophy. Other processes contained branching cisterns; still others were filled with electron-dense masses and amorphous vesicles. The overall ultrastructural appearance of variant CJD was similar to that of FFI cerebral cortex, except for a much higher number of cellular processes containing TVS. We detected TVS in the majority of sCJD cases that, in addition to typical spongiform change and robust astrocytic reaction, showed widespread neuritic and synaptic degeneration and autophagic vacuoles. We conclude that TVS are readily found in FFI, vCJD and sCJD and that widespread neuritic degeneration is a part of ultrastructural pathology in prion diseases.

Molecular heterogeneity of meningioma with INI1 mutation.

BACKGROUND: INI1 (hSNF5) mutations are linked to rhabdoid tumours, but mutations in meningiomas with hot spot mutations in position 377 have also been reported. AIMS: To analyse the INI1 gene in meningioma. METHODS: Exons 1, 4, 5, and 9 of the INI1 gene were analysed by the polymerase chain reaction and direct sequencing in 80 meningiomas. For all cases, western blotting of the INI1 protein was performed. RESULTS: Only one of the 80 samples showed a cytosine insertion in codon 376. This mutation changed the open reading frame in almost the whole exon 9 and resulted in a longer hSNF5 protein. Complex analysis of the above described tumour sample by western blotting, DNA sequencing, and loss of heterozygosity (LOH) analysis showed that this particular meningioma consisted of heterogeneic cellular components. One of these components had a mutated INI1 gene, whereas in the other component INI1 was intact. CONCLUSIONS: INI1 mutation is a rare event in the molecular pathology of meningiomas. It is possible for the INI1 gene to be mutated in only a proportion of meningioma cells.

Ultrastructural changes in the optic nerves of rodents with experimental Creutzfeldt-Jakob Disease (CJD), Gerstmann-Sträussler-Scheinker disease (GSS) or scrapie.

This report describes the ultrastructural changes in the optic nerves of (1) hamsters infected with the Echigo-1 strain of Creutzfeldt-Jakob disease (CJD), (2) hamsters infected with the 263K or 22C-H strain of scrapie, and (3) mice infected with the Fujisaki strain of Gerstmann-Sträussler-Scheinker disease (GSS). Vacuolation of myelinated fibres was present in the myelin sheaths, with splitting of myelin lamellae. These vacuoles contained typical secondary vacuoles and curled membrane fragments. Myelinated fibre vacuolation was also accompanied by an exuberant cellular reaction consisting of macrophages containing numerous mitochondria, abundant rough endoplasmic reticulum, and secondary lysosomes filled with digested myelin debris and other electron-dense material. Within macrophages, myelin fragments undergoing active digestion, lyre-like bodies and paracrystalline inclusions were frequently noted. Astrocytes and their processes were prominent; glial filaments and many mitochondria were readily detected. Proliferation of inner mesaxons was observed. Cross-sectional profiles of innumerable myelinated fibres contained membranous organelles continuous with the inner lamellae of the oligodendroglial cells. The proliferations of inner mesaxons formed whorls and loops, and intrusion of the membranous tongue of the inner mesaxon into the axoplasm was occasionally observed; dystrophic neurites were relatively numerous. In mice infected with the Fujisaki strain of GSS, fibres had undergone demyelination with stripping of the myelin lamellae, while others showed vesicular myelin degeneration.

[Pregnancy in patients with end-stage renal failure on maintenance dialysis: case reports]. / Ciaza u pacjentek przewlekle dializowanych z powodu terminalnej niewydolnosci nerek--opisy przypadków.

Pregnancy in women with end-stage renal failure on maintenance dialysis is rare, and the chance of successful delivery is relatively low. In this paper we present two cases of women who conceived just prior to initiation of renal replacement therapy and the pregnancy was terminated successfully already on chronic dialysis treatment. The special attention was paid on the necessity of multi-disciplinary collaboration and the need for changes in regular dialysis schedule as the conditions crucial for successful delivery. In summary, the review of current literature dealing with mentioned problem was done.

Prognostic significance of apoptosis in laryngeal cancer. A quantitative immunomorphological study.

The main purpose of the study was to investigate the prognostic significance of apoptosis of cancer cells and to examine the relationship between apoptosis and morphological markers of the host immune response in laryngeal cancer. Apoptotic tumour cells, detected by the TUNEL technique, expression of HLA DR (an antigen belonging to human leukocyte-associated antigens class II) in cancer cells, the number of tumour infiltrating T cells (CD45RO+ cells), B cells (CD20+ cells), macrophages (CD68+ cells) and mast cells as well as the mitotic index were investigated in 28 laryngeal cancers. Sections were studied morphometrically using image analysis. Significant inverse correlations between the number of apoptotic tumour cells and HLA DR-positive tumour cells as well as between the number of apoptotic tumour cells and the number of CD45RO+ cells at the tumour periphery were observed. Analysis of survival showed that patients with high rates of apoptosis had significantly worse prognosis as compared to patients with low apoptotic rates. Differences in HLA DR expression and numbers of CD45RO+ cells were also found between groups of patients with high and low numbers of apoptotic cells. Whether these findings are due to immunosuppressive effects of apoptosis needs further investigation.

[A rare case of neurofibroma in the parapharyngeal space, pharynx, skull base and neck area]. / Rzadki przypadek neurofibroma przestrzeni przygardlowej, gardla i podstawy czaszki.

Authors presented a rare case of neurofibroma involving skull base, parapharyngeal space and neck. A special emphasis was put on the preoperative diagnostic procedures and diagnostic complications as well as the complications due to the type of the operated tumor and its location.

HLA-DR expression is a significant prognostic factor in laryngeal cancer. A morphometric study.

HLA-DR antigen expression in tumour cells was investigated in 68 laryngeal carcinomas. The tissue sections were studied by a morphometric method using a computer image analysis system. Although the highest number of HLA-DR-positive cells was observed in the groups with the lowest clinical stage, these differences did not reach statistical significance. Analysis of survival showed a significantly different survival time of patients classified as HLA-DR positive (10% and more HLA-DR+tumour cells) compared to those classified as HLA-DR negative. There were no significant differences in survival times between patients with glottic and extraglottic cancers, or between patients with various grades of histological malignancy.

Quantitative analysis of HLA DR expression and lymphocytic infiltrate in laryngeal cancer including clinical and morphological correlations.

HLA DR antigen expression in tumor cells and lymphocytic infiltrates at the tumour periphery were investigated in 68 laryngeal carcinomas. The tissue sections were studied by a quantitative method using computer image analysis system. The highest number of CD45RO+ cells was observed in the groups of the lowest clinical stage. There were statistically significant differences between the groups in the number of T cells, whereas no differences were found in HLA DR+ cancer cell number, B cell number or mitosis count. Statistical analysis revealed a significant correlation between T-cell number at the periphery of the tumour and the number of cancer cells showing HLA DR expression. The results of the analysis, using Cox proportional hazard model with selected factors, showed that only HLA DR expression in cancer cells had a significant effect on survival, whereas the number of CD45RO+ and CD20+ cells, histological grade and mitosis count did not contribute further to prognostic information. To our knowledge, this is the first time that HLA DR expression and its correlation with lymphocytic infiltration have been shown by a quantitative method in laryngeal cancer.

Glomerular and interstitial renal findings in patients with liver cirrhosis with normal renal function. The histomorphometric study.

Seventeen kidney autopsy tissue sections from patients with cirrhosis of the liver were quantitatively examined and compared with fifteen cases of normal controls. The study was limited only to cases with normal renal function and without macroscopic renal changes. Morphometric investigations were performed by means of a computer image analysis system. The study revealed that total glomerular area, total number of glomerular cells per total glomerular area, mesangium (percentage of total glomerular area), and number of sclerotic glomeruli (percentage of total glomeruli) were significantly increased in cirrhotic patients. There was no significant increase in relative interstitial volume and in total glomerular cell per unit of glomerular area in comparison with controls. The results indicate that, despite the lack of renal clinical and laboratory manifestation, cirrhosis of the liver can be accompanied by a variety of glomerular disorders. These abnormalities consist of an enlargement of the glomeruli, an increase in glomerular cellularity proportional to the increase in glomerular size, thickening in mesangial matrix, and increased glomerulo-sclerotic lesions.

Acute myelofibrosis in children: report on two cases.

In our report, myelofibrosis in children is discussed and two cases of acutely developing myelofibrosis in association with acute megakaryoblastic leukaemia (M7) are presented. In the first case (girl, 34 months), it was acute myelofibrosis of hypocellular marrow. Diagnosis of M7 was confirmed by positive reaction of blasts from peripheral blood with CD42 and CD61 monoclonal antibodies. In the other child (girl, 5 years old), Ph1(+) chronic myeloid leukaemia diagnosed 22 months earlier transformed to M7. Similar to the first case, no marrow aspirate could be obtained and the diagnosis of M7 was made by the bone marrow histology that showed the presence of grossly fibrosed, hypercellular marrow with a large number of dysplastic, maturing megakaryocytes. Neither of the children had Down's syndrome. Although according to FAB classification both cases represent the same haematological entity, their clinical and histopathological presentations are very different.

[Endometriosis of the large intestine]. / Endometrioza jelita grubego.

Two cases of large intestine endometriosis are presented. The disease was diagnosed during histological examination of samples taken during surgery. Clinically one case was diagnosed as Crohn's disease, while the second as cancer of the large intestine. The authors suggest, that an extent of surgery for, tumours of the large intestine should be carefully planned, specially if the tissue specimen was not examined histologically earlier.