Globular glial tauopathy caused by MAPT P301T mutation: clinical and neuropathological findings.

OBJECTIVE: To describe the clinical, biochemical, and neuropathological findings of an autosomal dominant globular glial tauopathy caused by the P301T mutation at the MAPT gene. METHODS: Five patients from two unrelated pedigrees underwent clinical evaluation. Genetic analysis, brain pathological examination, and biochemical analysis of tau were performed. RESULTS: The patients studied were 3 men and 2 women with a mean age at onset of 52.2 years and mean disease duration of 5.2 years. Three patients presented a corticobasal syndrome, one patient an asymmetric pyramidal syndrome compatible with primary lateral sclerosis, and one patient a frontotemporal dementia. In both pedigrees (4 patients) Sanger sequencing showed the p.P301T mutation in exon 10 of the MAPT gene. Neuropathological findings consisted of atrophy of frontal and temporal lobes with marked spongiosis and astrogliosis, and abundant phosphorylated tau protein deposits in the frontal and temporal cortex, limbic area, basal ganglia, and brain stem. The most striking finding was the presence of oligodendroglial 4R phospho-tau globular positive inclusions in the white matter and cortex. Globose-type neurofibrillary neuronal tangles, and in particular astrocytic globular inclusions and coarse tufts, were present in the grey matter. Biochemical analysis of sarkosyl-insoluble fractions revealed two tau bands of 64 and 68 kDa and case-dependent bands of lower molecular weight. CONCLUSION: This is the first pathological and biochemical study of the MAPT p.P301T mutation showing variable clinical manifestation and neuropathological phenotype of globular glial tauopathy not only among different families but also within families.

Multiprotein deposits in neurodegenerative disorders: our experience in the tissue brain bank of Navarra.

The prevalence of neurodegenerative disorders increases dramatically with advancing age. Although in recent decades the study of many neurodegenerative disorders has evolved greatly, the concept of neurodegeneration still remains elusive. Although neurodegenerative disorders are classified according to the major components of protein deposits, coexpression of several abnormal proteins in the brain tissue is more common than that was previously thought. The aim of this report is to describe the type of protein deposits found in brains with neuropathological diagnosis of neurodegenerative disease. The report shows the experience obtained in the Brain Bank of Navarra (Spain). The target population for this retrospective descriptive study comprised 178 brains autopsied in the "Hospital of Navarra" in Pamplona between 1994 and 2004 and 201 brains donated to the Brain Bank of Pamplona between 2004 and 2009. The diagnosis of the 201 brains from the Brain Bank was 62 (30.8%) Alzheimer's disease (AD), 43 (21.3%) multiprotein deposit, 31 (15.4%) α-synucleinopathies, 31 (15.4%) frontotemporal lobar degeneration (FTLD), 17 (8.4%) tauopathies, 9 (4.4%) prion disease, 6 (2.9%) vascular dementia (VD), and 2 (0.9%) Huntington's disease. Among the 43 cases with multiprotein deposits, we found 35 brains with deposits of 3 proteins (tau, ß-amyloid, and α-synuclein). In these two series of brains, the high incidence of deposition of multiple proteins in neurodegenerative disorders is shown. Our results are in agreement with previous findings showing that tau, ß-amyloid, and α-synuclein are the proteins most frequently deposited together.

[The role of diffusion-weighted imaging in the evaluation of meningiomas: radio-pathologic correlation]. / Papel de la difusión en la evaluación de los meningiomas: correlación radiopatológica.

OBJECTIVE: To study the behavior of meningiomas in diffusion-weighted sequences and the correlation of these findings with the histological findings. MATERIAL AND METHODS: We prospectively included all patients operated on for meningiomas at our hospital during two years. We studied 30 meningiomas in 28 patients aged 31 to 85 years old. All patients underwent MRI prior to surgery, including diffusion-weighted sequences, in a 1.5 T scanner. We evaluated the signal intensity in T2-weighted images, diffusion-weighted images (b=1,000), and apparent diffusion coefficient (ADC) maps within the tumor and in the parietal white matter as a reference. In the histological study, cellularity, proliferation index, histological grade, and cerebral invasion were evaluated. RESULTS: Of the 30 meningiomas, 22 were World Health Organization (WHO) grade I and 8 were atypical or WHO grade II. The overall mean value of the ADC was 89.19+/-13.95x10(-3) mm2/s; the mean ADC value was 82+/-13.69x10(-3) mm2/s in the atypical group and 92.21+/-13.21x10(-3) mm2/s in the typical group. No statistically significant differences were found between the 2 groups. Two subtypes of typical meningiomas, secretory and angiomatous meningiomas, had the highest values in the ADC maps. In the histological analysis, there was a significant association between tumor cellularity and the signal in the ADC map. CONCLUSION: Meningiomas show moderately restricted diffusion. The signal on the ADC map is associated with tumor cellularity but we have not demonstrated its usefulness for predicting the histological grade.

A patient with MV2 subtype of sporadic Creutzfeldt-Jakob disease and atypical clinical presentation.

We report the case of a 71-year-old woman with progressive dementia over the course of 4 years, characterized by prominent pyramidal signs and by the lack of ataxia and other cerebellar signs. Creutzfeldt-Jakob disease (CJD) was not suspected during the patient's life. Autopsy brain tissue showed severe spongiform encephalopathy with kuru-like, but not florid, plaques in neocortex and cerebellum. Massive synaptic diffuse and plaque-like PrP(Sc) deposition was found in the cerebral cortex, striatum, cerebellum and brainstem. Genetic analysis revealed no PRNP gene mutations and methionine/valine heterozygosity (MV) at codon 129. The pathogenic scrapie prion protein (PrP(Sc)) pattern detected by Western blot was Type 2. However, this pattern showed a single unglycosylated band in contrast to the doublet described for MV2 subtype of sCJD with kuru plaques. In summary, this is an autopsy case report of a particular presentation of MV2 subtype of sCJD.

Danon disease: a novel Lamp-2 gene mutation in a family with four affected members.

This is a report of a family with four members affected with Danon disease and variable clinical presentations, including cardiomyopathy, skeletal muscle pathology, and hepatopathy. Analysis by electron microscopy of the quadriceps muscle from the proband and his brother showed abnormal mitochondria, and immunohistochemistry revealed no expression of LAMP-2 protein. This defect is due to a yet undescribed mutation located at the second nucleotide in the intron 8 of the Lamp-2 gene (c.1093+2 T>A) that generated exon 8 skipping confirmed at RNA level in the proband.

[Sleep disorders in prion diseases]. / Patología del sueño en las enfermedades priónicas.

Prion diseases are a group of encephalopathies with neurodegenerative changes caused by an altered protein named prion whose characteristic datum is transmissibility. In most cases they occur in a sporadic form although a group of them are familial associated with mutations in the gene of the prion protein. Genetic polymorphism seems to determine the different family variants. One of the most enigmatic and unusual is Fatal Familial Insomnia (FFI), a hereditary disorder characterised by loss of physiological sleep with oneiric stupor, autonomic and motor hyperactivity, and motor anomalies. The polysomnography of this entity reflects an inability to produce the physiological pattern of NREM and REM sleep, as well as hormonal and vegetative circadian fluctuations; the transition from wakefulness to sleep is markedly altered with the early disappearance sleep spindles. The hypothesis of the origin of these disorders is thalamic neuronal loss, especially in the anterior and dorsomedial nuclei, described in the neuropathology of these patients; besides PET reveals hypofunction of thalamic nuclei, centres responsible for controlling wakefulness-sleep. In Creutzfeldt-Jakob disease the wake-sleep disorders are not considered characteristic; nonetheless, frequent alterations have been found in the electroencephalographic registers of sleep. Besides thalamic neurodegeneration, there could be common etiopathogenic mechanisms in prion diseases in relation to the biological function of the prion protein.

High promoter hypermethylation frequency of p14/ARF in supratentorial PNET but not in medulloblastoma.

AIMS: Medulloblastoma (MB) is the most common primitive neuroectodermal tumour (PNET) of the central nervous system. Although supratentorial PNET (sPNET) and MB are histologically similar, their clinical behaviour differs, sPNET being more aggressive than MB. The aim of this study was to determine whether sPNET and MB are genetically different entities. METHODS AND RESULTS: We investigated 32 PNET primary tumour samples (23 MB and nine sPNET) and four PNET cell lines, for the presence of CDKN2A homozygous deletions at exon 1-alpha of p16/INK4 and exon 1-beta of p14/ARF, and promoter hypermethylation of both genes. No homozygous deletion of either p16/INK4 or p14/ARF was demonstrated in any of the PNET primary tumour samples. Methylation of p16/INK4 was found in one of six sPNET and in one of 23 MB, while p14/ARF methylation was observed in three of six sPNET and in three of 21 MB. No methylation of p16/INK4 or p14/ARF was found in any of the PNET cell lines analysed. The three MB cell lines did not show p16/INK4 expression, and only the MB Daoy cell line (homozygously deleted at CDKN2A) presented loss of p14/ARF expression. CONCLUSIONS: Our results in this limited series of central PNET show that p14/ARF is frequently involved in PNET carcinogenesis, with a higher frequency, but not statistically significant, for sPNET than for MB.

Genetic heterogeneity in supratentorial and infratentorial primitive neuroectodermal tumours of the central nervous system.

AIMS: Medulloblastoma (MB), a kind of infratentorial primitive neuroectodermal tumour (PNET), is the most frequent malignant brain tumour in childhood. In contrast, supratentorial PNET (sPNET) are very infrequent tumours, but they are histologically similar to MB, although they present a worse clinical outcome. We investigated the differences in genetic abnormalities between sPNET and MB. METHODS AND RESULTS: We analysed 20 central PNET (14 MB and six sPNET) by conventional comparative genomic hybridization (CGH) in order to determine whether a different genetic profile for each tumour exists. Isochromosome 17q was detected in four of the 14 MB cases, but not in any sPNET. Gains at 17q and 7 happened more frequently in MB, and those at 1q in sPNET. Losses at chromosome 10 were detected only in MB, while losses at 16p and 19p happened more frequently in sPNET. A new amplification site, on 4q12, was detected in two MB. CONCLUSIONS: Central PNET are a heterogeneous group of tumours from the genetic point of view. The present and previous data, together with further results from larger series, might contribute to the establishment of specific treatments for supratentorial and infratentorial PNET.

[Primary CNS Lymphoma: bibliographical review and experience at the Hospital of Navarre in the last 5 years (2000-2004)]. / Linfoma cerebral primario: revisión bibliográfica y experiencia en el Hospital de Navarra en los últimos 5 años (2000-2004).

Primary cerebral lymphoma (Primary CNS Lymphoma, PCNSL) is an aggressive non-Hodgkin lymphoma that originates in the central nervous system without evidence of lymphoma in any other localization at the time of diagnosis. Primary cerebral lymphomas are less well-known and are characterized than their homologues the systemic lymphomas, as they are an entity whose frequency was scarce until a few decades ago. However, the great rise in incidence that this pathology has undergone over the last three decades, and which is still unexplained, makes more studies necessary to better understand the etiopathology of this entity. Thanks to the new systems of treatment, the prognosis of this pathology has improved significantly in recent years. Nonetheless, treatment of primary cerebral lymphoma continues to give rise to numerous controversies at present due to its high neurotoxicity in patients over 60 years of age, a group of patients frequently affected by this pathology. To resolve these and other questions it is necessary to deep in the study of primary cerebral lymphoma and to carry out high quality clinical trials.

Improvement of the methylation specific PCR technical conditions for the detection of p16 promoter hypermethylation in small amounts of tumor DNA.

Methylation specific PCR (MSP) is a technique that enables the detection of hypermethylation at a CpG island. The objective of this study is the introduction of small modifications to the MSP technique to make it more suitable for the study of promoter hypermethylation at tumor suppressor genes whenever there is a shortage of material available for study. This commonly happens in the case of using archival material from the Pathology departments. Tumor DNA was extracted from a collection of 40 fresh-frozen soft tissue sarcomas and 19 paraffin-embedded PNETs (primitive neuroectodermal tumors). The MSP technique was performed to detect hypermethylation at the p16 promoter. Also, blood genomic DNA was mixed with herry sperm genomic DNA as a carrier, in nine different combinations, in order to test for the best conditions that could produce MSP bands even when low amounts of genomic tumor DNA is available for study. We demonstrate the benefit of using herry sperm carrier DNA up to 10 microg together with small quantities of tumor DNA. This result will facilitate the incorporation of paraffin-embedded samples for study of promoter hypermethylation at tumor suppressor genes. Other technical conditions for the MSP technique are also studied.

Single large-scale mitochondrial DNA deletion in a patient with encephalopathy, cardiomyopathy, and prominent intestinal pseudo-obstruction.

We studied a 62 year-old woman with a clinical phenotype characterized by encephalopathy, restrictive cardiomyopathy, and prominent intestinal pseudo-obstruction. Muscle morphology showed ragged red fibres with ultrastructurally abnormal mitochondrial whereas muscle respiratory chain was normal. Molecular genetics revealed the 'common deletion' in mtDNA, which represented 40% of total mtDNA. These data expand and confirm the wide clinical spectrum of mitochondrial disorders associated with single large-scale mtDNA deletions.

Immunopathogenesis of human gastrointestinal infection by Anisakis simplex.

BACKGROUND: Anisakis simplex is a parasite of fish, and in the case of human infestation, it should be considered as a possible cause of gastrointestinal disease, especially in countries where raw or undercooked fish is a frequent food. Clinical features of anisakiasis may simulate acute abdominal pain, such as that found in patients with gastric ulcers, appendicitis, and Crohn's disease. Furthermore, many cases of anisakiasis are diagnosed as eosinophilic gastroenteritis, which is a broad term for a specific disease. OBJECTIVE: The purpose of this study was to investigate the immunopathogenesis of human gastrointestinal infestation by A simplex. METHODS: Thirteen intestinal biopsy specimens from patients with anisakiasis were analyzed for the presence of messenger (m)RNA for different cytokines and inflammatory mediators by RT-PCR. Specific IgE, eosinophil cationic protein, eosinophil protein X, and tryptase levels were measured in each patient's serum. Also, cell cultures were set up with lymphocytes from some patients and stimulated in vitro with Anisakis and Ascaris antigens. RESULTS: We performed immunologic phenotyping in 13 patients. All patients underwent biopsy after emergency surgery caused by episodes of acute abdominal pain. In all cases inflammatory infiltrate composed of eosinophils and lymphocytes was found in the intestinal wall. We demonstrated that after infestation, a T(H2)-type immune response occurred. Also, major basic protein, nitric oxide, and eotaxin were found in the tissue, and eosinophil cationic protein and eosinophil protein X levels were elevated in sera. CONCLUSION: These data and in vitro lymphocyte cultures indicate that a T(H2) mechanism plays an important role in the inflammatory infiltrate produced by the anchorage of parasites in the gastrointestinal wall.

Eosinophilic gastroenteritis and Anisakis.

BACKGROUND: The differential diagnosis of eosinophilic gastroenteritis (EG) includes, among other diseases, parasitic infections such as anisakiasis, which has acquired worldwide importance. METHODS: We reviewed all patients referred to our allergy service who had been diagnosed as having primary EG to determine the possible role of Anisakis simplex in the etiopathology of the disease. All patients (n = 10) were studied and diagnosed as having primary EG between 1989 and 1996, inclusive. Two different groups of subjects were used as controls: group A (149 subjects without digestive disorder) and group B (10 subjects with digestive disorder different from EG). Cutaneous prick tests were performed with the main foods, aeroallergens, and commercial extract of A. simplex. Total and specific serum IgE was measured in all patients. Gastric or gut histologic specimens were re-examined in five cases. RESULTS: Peripheral eosinophilia was detected in 40% of the patients with EG, and sensitization to A. simplex was detected in 80% of these. In both control groups, the rate of sensitization to A. simplex was 10%. Sensitization to A. simplex in EG patients with respect to control groups A and B showed odds ratios (OR) of 36 and 40, respectively. In one case, serialization of the histologic section allowed us to observe a whole Anisakis larva. CONCLUSIONS: Immunologic methods to detect specific antibodies against Anisakis should be used routinely before diagnosing EG as primary disorder. Preventive measures are of capital importance.

[Lewy body disease: first figures on its frequency in Spain]. / Enfermedad con cuerpos de Lewy difusos: primeros datos de frecuencia en España.

Lewy body disease (LBD) as a separate nosologic entity causing dementia in the elderly is being firmly established. To know its prevalence and characterization, we reviewed 549 consecutive autopsied brains in our Department. The age of death was 60 years or older in 391 subjects. Immunohistochemical staining with ubiquitin antibodies facilitated the identification of LB. Their specific density was measured (number per 100xfield) following a protocol in the predilection neocortical sites, entorhinal cortex, hippocampal gyrus, diencephalon and brainstem. We assessed the clinical features according to LB findings. Twelve brains (2.1%) had neocortical LB. Nine of them were diagnosed as diffuse Lewy body disease (DLBD). One more brain had nigral and neocortical LB leading to a pathological diagnosis of PD. In the remaining 2 cases, the finding of neocortical LB seems to be either incidental or asymptomatic or preclinical. Cognitive decline was mild to moderate in all subjects which had neocortical LB in 4 or more areas. However, the density of these LB does not correlate with the severity of dementia. Dementia was associated with minor parkinsonian symptoms and psychiatric features in the most of patients with DLBD. Thirty eight cases of the 391 (9.7%) older than 60 years in these series had been clinically diagnosed as senile dementia. Using accepted neuropathologic criteria, diagnoses were AD (63.1%), DLBD (21.05%) and vascular dementia (13.1%). These observation suggest that on consultant diagnosis of senile dementia, DLBD must be taken in account.

[Preliminary results of the study of neuronal death and the expression of bcl-2 protein in Alzheimer's disease]. / Resultados preliminares del estudio de la muerte neuronal y de la expresión de la proteína bcl-2 en la enfermedad de Alzheimer.

Alzheimer's disease (AD) is a degenerative central nervous system disorder where beside the histopathologic features of senile plaques and neurofibrillary tangles there is an important neuronal loss. It has been suggested that this neuronal death occurs via an apoptotic mechanism. Recognition of apoptotic cells is possible by an in situ end-labeling technique which identify the 3'-OH termini of DNA strands breaks through the incorporation of labeled nucleotides with the enzyme terminal-deoxinucleotidyl transferase (Tdt). We have applied this technique and high densities of apoptotic cells were found in 5 AD brains compared to 5 age-matched normal samples. We studied by immunohistochemical analyses the expression of the antiapoptotic protein bcl-2. We have not found neuronal bcl-2 immunoreactivity and we found an increased expression of bcl-2 by astrocytes compared to controls, this fact may aid glial survival or may have a deleterious effect on neuronal viability.