The "Brescia panel" (Claudin-4 and BRCA-associated protein 1) in the differential diagnosis of mesotheliomas with epithelioid features versus metastatic carcinomas.

BACKGROUND: The distinction between mesothelioma with epithelioid features and metastatic carcinoma may be challenging, particularly on cytology. A novel 2-hit Claudin-4 and BRCA-associated protein 1 (BAP1) panel was investigated. METHODS: The objective of this study was to determine the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the panel on cytology from pleural effusions and matched biopsies, including 49 malignant mesotheliomas on cytology with 43 matched biopsies, 49 normal/reactive mesothelial proliferations, and 49 pleural metastatic carcinomas from different primaries with 21 matched pleural biopsies. The diagnostic role of the 4 categories obtained by crossing the immunostaining results was analyzed. RESULTS: Claudin-4 strongly stained all metastatic carcinomas and tested completely negative in normal mesothelium, benign reactive mesothelial hyperplasia, and malignant mesothelioma. All normal and benign mesothelial proliferations and all carcinomas except 1 were immunoreactive for BAP1, whereas BAP1 loss was observed in 88% of malignant mesotheliomas. The expression of Claudin-4 alone excluded all benign and malignant mesothelial growth, consistently characterizing all metastatic carcinomas. Double negativity was evident in all malignant mesotheliomas, and double positivity was observed in all metastatic carcinomas. BAP1-positive/Claudin-4-negative status was observed only in malignant mesotheliomas and benign mesothelial proliferations. A single metastatic anal squamous cell carcinoma had BAP1-negative/Claudin-4-positive staining. CONCLUSIONS: Claudin-4 expression was completely specific and sensitive for metastatic carcinoma, excluding mesothelial proliferations. BAP1 staining characterized 98% of metastatic carcinomas and 100% of benign mesothelial proliferations, whereas negativity was observed almost exclusively in mesotheliomas. This 2-hit panel is probably the best compromise for differentiating malignant mesothelioma and metastatic carcinoma on either cytology or biopsy specimens.

Genetic variation in APOE, GRN, and TP53 are phenotype modifiers in frontotemporal dementia.

Frontotemporal dementia (FTD) is a clinical, genetic, and pathologic heterogeneous group of neurodegenerative diseases. In this study, we investigated the role of APOƐ4, rs5848 in GRN, and rs1042522 in TP53 gene as disease risk factors and/or phenotype modifiers in 440 FTD patients, including 175 C9orf72 expansion carriers. We found that the C9orf72 expansion carriers showing an earlier age at onset (p < 0.001). Among the clinical groups, the FTD-MND (motoneuron disease) showed the lowest survival (hazard ratio [HR] = 4.12), and the progressive nonfluent aphasia group showed the highest onset age (p = 0.03). In our cohort, the rs1042522 in TP53 was associated with disease onset (p = 0.02) and survival (HR = 1.73) and rs5848 GRN with a significantly shorter survival in CC homozygous patients (HR = 1.98). The frequency of APOƐ4 carriers was significantly increased in the C9orf72 noncarriers (p = 0.022). Although validation of our findings is necessary, our results suggest that TP53, GRN, and APOE genes may act as phenotype modifiers in FTD and should be considered in future clinical trials.

From beta amyloid to altered proteostasis in Alzheimer's disease.

Alzheimer's disease (AD) is an age related neurodegenerative disorder causing severe disability and important socio-economic burden, but with no cure available to date. To disentangle this puzzling disease genetic studies represented an important way for the comprehension of pathogenic mechanisms. Abnormal processing and accumulation of amyloid-ß peptide (Aß) has been considered the main cause and trigger factor of the disease. The amyloid cascade theory has fallen into crisis because the failure of several anti-amyloid drugs trials and because of the simple equation AD = abnormal Aß deposition is not always the case. We now know that multiple neurodegenerative diseases share common pathogenic mechanisms leading to accumulation of misfolded protein species. Genome Wide Association studies (GWAS) led to the identification of large numbers of DNA common variants (SNPs) distributed on different chromosomes and modulating the Alzheimer's risk. GWAS genes fall into several common pathways such as immune system and neuroinflammation, lipid metabolism, synaptic dysfunction and endocytosis, all of them addressing to novel routes for different pathogenic mechanisms. Other hints could be derived from epidemiological and experimental studies showing some lifestyles may have a major role in the pathogenesis of many age-associated diseases by modifying cell metabolism, proteostasis and microglia mediated neuroinflammation.

Association of the variant Cys139Arg at GRN gene to the clinical spectrum of frontotemporal lobar degeneration.

BACKGROUND: Progranulin protein (PGRN) is a cysteine-rich growth factor encoded by the progranulin gene (GRN). PGRN mutations were identified in patients with frontotemporal lobar degeneration (FTLD) and recently its role as risk factor has been described in patients with probable Alzheimer's disease (AD). To date, more than 100 genetic variants in GRN gene have been described and the pathogenic nature is still unclear for almost 36% of them. OBJECTIVE: Here, we describe three clinical cases carrying the PGRN variation Cys139Arg in order to increase the knowledge on the association of this variant to the clinical spectrum of FTLD. METHODS: The genetic analysis was performed using high resolution melting analysis. The Human Progranulin ELISA Kit was used in order to determine PGRN expression levels in the plasma samples. RESULTS: The three patients carrying the genetic variation showed three final different clinical diagnosis, respectively behavioral frontotemporal dementia, semantic dementia, and corticobasal syndrome, thus underlining the clinical heterogeneity typically associated with GRN mutations. All cases shared similar plasma PGRN levels that resulted intermediate between those measured in controls and in GRN null mutation carriers, showing a partial reduction of the protein in plasma. Moreover, according to the bioinformatics software, the Cys139Arg variation causes a decreased stability of the structure of the protein. CONCLUSION: We describe three new patients affected by neurological syndromes included in the clinical spectrum of FTLD carrying the Cys139Arg genetic variant, thus suggesting a possible implication in the pathogenesis of FTLD.

First deep intronic mutation in the NOTCH3 gene in a family with late-onset CADASIL.

CADASIL is the most prominent inherited form of vascular dementia. The main clinical features include migraine with aura, stroke, mood disturbances, and cognitive decline, with a mid-life (30s-60s) adult onset. Genetic testing is the gold standard for the diagnosis. CADASIL is caused mostly by missense mutations in the NOTCH3 gene, invariably involving a cysteine residue. Only a couple of splice site mutations have been reported. In a few pathologically defined patients, genetic mutations remain unidentified. We report a family with late-onset CADASIL phenotype carrying a novel intronic deletion in the NOTCH3 gene (c.341-26_24delAAC). Transcript analysis revealed a splicing alteration, with the complete intron 3 retention. The insertion was in-frame and encoded an extra 25 amino acids, including 1 cysteine. This is the first report of an aberrant splicing event of the NOTCH3 gene associated with a mutation far away from the canonical splice site. Our finding suggests that the assays used to evaluate splicing should be mandatory in the diagnostic setting of genetically undefined CADASIL cases.

Epidemiology and genetics of frontotemporal dementia: a door-to-door survey in southern Italy.

The objectives of this study were to estimate frontotemporal dementia (FTD) prevalence, identify FTD-related mutations, and correlate FTD phenotype with mutations in a southern Italian population. The study population consisted of subjects ≥ 50 years of age residing in the Community of Biv. on January 1, 2004, and a door-to-door 2-phase design was used. Genetic and biochemical analyses were done on samples collected from 32 patients. Prevalence rates were 0.6 for Alzheimer's disease, 0.4 for vascular dementia (VD), 3.5 for FTD, 0.2 for Parkinson dementia, and 1.2 for unspecified dementia. Three GRN (1 known and 2 novel) mutations with reduced plasma protein levels were found associated to 3 distinct phenotypes (behavioral, affective, and delirious type). We report an unusually high FTD prevalence in the investigated population, but a low prevalence of Alzheimer's disease. We confirm the heterogeneity of FTD phenotype associated with different GRN mutations.

Novel PSEN1 and PGRN mutations in early-onset familial frontotemporal dementia.

BACKGROUND: Frontotemporal dementia is a clinically and genetically heterogeneous syndrome. Mutations in two genes, Microtubule Associated Protein Tau (MAPT) and Progranulin (PGRN), and rarely Presenilin mutations, have been causally linked to this disorder. OBJECTIVE: To investigate the presence of PGRN, PSEN1, PSEN2 and APP mutations in a group of familial early-onset frontotemporal dementia (f-EOFTD) patients negative for MAPT gene mutations. SUBJECTS AND METHODS: We prospectively studied 17 unrelated subjects diagnosed with f-EOFTD (one case neuropathologically confirmed as FTD-Ub+). Among these subjects eight belonged to eight autosomal dominant families unrelated to each other, and nine had at least one first degree relative affected by dementia. RESULTS: We identified two novel heterozygous mutations in two unrelated patients, Cys139Arg in the PGRN gene and Val412Ile in the PSEN1 gene. CONCLUSIONS: Early-onset f-FTD remains a heterogeneous disorder from a genetic point of view. PGRN mutation frequency was low in our sample. The presence of a novel PSEN1 mutation suggests that presenilin molecular studies should be performed when screening for MAPT and PGRN genes is negative.

A acupuntura como terapêutica da síndrome do intestino irritável / Acupuncture as treatment for irritable bowel syndrome

Introdução: A Síndrome do Intestino Irritável (SII) é uma das mais comuns desordens gastrointestinais dasociedade ocidental, afetando cerca de 15% da população, especialmente adultos jovens. Tratamentos efetivostêm permanecidos indefinidos, já que muitos pacientes não conseguem o alívio adequado para os seus sintomascom o uso das terapias convencionais.Discussão: Tratamentos alternativos, como a acupuntura, estão tornando-se cada vez mais populares. Dentre as doenças do trato gastrointestinal, o uso mais comum dessas terapias é na SII. A resposta-placebo nos casos de SII pode chegar a 80% em ensaios simples,com uma resposta média de aproximadamente 40%, sendo que os preditores de uma resposta-placebo nãosão conhecidos e requerem análise de dados individuais tanto quanto a metanálise de ensaios publicados. Schneider et al. (Gut 2006;55:649-4), no maior estudo controlado e randomizado sobre o assunto, não achou diferença entre acupuntura e falsa acupuntura em relação aos principais desfechos pesquisados. Conclusão: Os efeitos da acupuntura na SII parecemser devidos ao efeito placebo. A razão para essa alta resposta pode ser atribuída às intensas particularidadespessoais durante o tratamento com acupuntura, combinada com a capacidade do paciente de relaxar emdeterminados ambientes calmos.

Introduction: Irritable bowel syndrome (IBS) is one of the most common disorders of western society; hence15% of its population is affected, especially young adults. Effective treatment has been remaining undefined,since many patients fail to reach adequate relief to their symptoms through conventional therapies.Discussion: Complementary therapies like acupuncture are growing in popularity. The most common use ofthese therapies in gastrointestinal diseases is the IBS. Placebo-effect efficacy in IBS can reach 80% in simplestudies, with an average response of nearly 40%. The predictors for placebo-effect efficacy are unknown and need analysis of individual data as much as of methanalysis of published studies. Schneider et al. (Gut 2006;55:649-4), at the largest controlled randomized study on the subject, found no difference between real and false acupuncture on the researched outcomes. Conclusion: Effects of acupuncture in IBS seem tobe due to placebo-effect. The reason for such high response can be given to the personal particularities of acupuncturetherapy, combined with the patient’s capacity of relaxation on calm environments.

Relação entre o tamanho da hérnia hiatal e a gravidade da esofagite de refluxo / Relationship between hiatal hernia size and reflux esophagitis severity

Objetivo: Avaliar por parâmetros endoscópicos a relação do tamanho da hérnia hiatal (HH) com a gravidade da esofagite de refluxo (ER). Métodos: Foram revistas 3.001 endoscopias digestivas altas. Utilizou-se como critério de inclusão a presença concomitante de HH e ER. Para o diagnóstico endoscópico da HH considerou-se a elevação da linha Z a 2cm ou mais acima do pinçamento diafragmático. Para a caracterização macroscópica da ER, foram adotadas as classificações de Los Angeles e Savary-Miller. Resultados: Os critérios de inclusão foram preenchidos por 408 pacientes. Os resultados foram reunidos em cinco grupos distintos segundo os critérios de Los Angeles e Savary-Miller para a ER. Comparando-se as médias dos tamanhos das hérnias hiatais, não houve diferença estatisticamente significante entre os grupos um (ER não erosiva) e dois (LA A/ SM I), que evidenciaram ER leve e HHs de pequeno volume (médias 2,6 e 2,8cm, respectivamente). Entre os grupos três (LA BI SM II) e quatro (LA CI SM III), que apresentaram ER moderada e HHs de volume moderado (médias 3,2 e 3,6cm, respectivamente), também não houve diferença estatística. O grupo cinco (LA DI SM IV), estatisticamente diferente dos demais (p < 0,001), apresentou HHs volumosas (média de 4,9cm) e ER grave. Conclusão: As ERs mais graves apresentaram HHs mais volumosas, sendo seu tamanho diretamente proporcional ao da gravidade da ER.

Hérnia Hiatal, Esôfago de Barrett e a Gravidade da Esofagite de Refluxo / Hiatal Hernia, BarrettÆs Esophagus and Reflux Esophagitis Severity

Contexto e Objetivo: Relação entre a hérnia hiatal e a doença do refluxo gastroesofágico (DRGE) ainda não é completamente compreendida. Não se tem certeza quanto à hérnia hiatal ser um fator de risco, causa ou perpetuação da DRGE. O objetivo do presente estudo é associar complicações endoscópicas da DRGE, como a esofagite de refluxo e o Esôfago de Barrett, à presença de hérnia hiatal. Método: Foram revisados 3001 resultados de endoscopias digestivas altas, de 2000 a 2005, sendo incluídos neste estudo 2817 desses prontuários. Foi utilizado neste estudo o diagnóstico endoscópico de esofagite de refluxo, hérnia hiatal e de Esôfago de Barrett. A análise estatística foi feita pelo teste do qui-quadrado. Resultados: Foi encontrada hérnia hiatal em 59,4% dos pacientes com esofagite de refluxo não-erosiva (OR = 10,2), 66,9% dos pacientes com esofagite LA A/SM I (OR = 13,8), 60,3% dos pacientes com LA B/SM II (OR = 10,5), 91,7% dos pacientes com LA C/SM III (OR = 76,8), 50% dos pacientes com LA D/SM IV (OR = 6,9) e 60,3% dos pacientes com Esôfago de Barrett (OR = 5,0). Todas as associações foram estatisticamente significativas (p < 0,001). Conclusões: A presença de hérnia hiatal em pacientes com DRGE foi um fator importante para o achado de esofagite de refluxo e de Esôfago de Barrett, aumentando substancialmente o risco de desenvolvimento dessas complicações da DRGE.

Context and Objective: Relationship between hiatal hernia and gastroesophageal reflux disease (GERD) has not yet been completely understood. ItÆs not fully known if hiatal hernia is a risk, causal or perpetuation factor for GERD. The aim of the present study was to successfully associate endoscopical complications of GERD, such as reflux esophagitis and BarrettÆs Esophagus, to the presence of hiatal hernia.Methods: It was revised 3001 result charts of upper gastrointestinal endoscopy, from 2000 to 2005, being included 2817 of these charts. It was used in this study endoscopical diagnosis of reflux esophagitis, hiatal hernia and BarrettÆs Esophagus. Statistics were given by chi-square test.Results: Hiatal hernia was found in 59,4% of patients with non-erosive reflux esophagitis (OR = 10,2), in 66,9% of LA A/SM I esophagitis patients (OR = 13,8), in 60,3% of LA B/SM II patients (OR = 10,5), in 91,7% of LA C/SM IV patients (OR = 76,8), in 50% of LA D/SM IV patients (OR = 6,9) and in 60,3% of BarrettÆs Esophagus/SM V patients (OR = 5,0). All associations were of statistic significance (p < 0.001). Conclusions: The presence of hiatal hernia in GERD in the present study was a fundamental factor in the finding of reflux esophagitis and BarrettÆs Esophagus, increasing substantially the risk for the development of such GERD complications.

Circulating levels of soluble receptor for advanced glycation end products in Alzheimer disease and vascular dementia.

BACKGROUND: The receptor for advanced glycation end products (RAGE) is a cell surface receptor that has been implicated in vascular disease and neurodegeneration. Low levels of its secreted isoform, soluble RAGE (sRAGE), have been regarded as a putative risk factor for atherosclerosis. In addition, administration of sRAGE has been shown to reduce development of cerebral beta-amyloidosis in an Alzheimer disease mouse model. OBJECTIVE: To investigate the role of sRAGE as a biological marker for Alzheimer disease and vascular dementia. DESIGN: Cross-sectional study of 152 patients with a clinical diagnosis of Alzheimer disease, 91 with vascular dementia and 161 control subjects. MAIN OUTCOME MEASURE: Plasma levels of sRAGE. RESULTS: Levels of sRAGE were significantly reduced in the plasma of patients with Alzheimer disease compared with that for those with either vascular dementia (P<.05) or with controls (P<.001). CONCLUSIONS: Patients with Alzheimer disease have reduced levels of sRAGE in plasma compared with patients with vascular dementia and controls. The striking reduction of circulating sRAGE in Alzheimer disease further supports a role for the RAGE axis in this clinical entity and requires further investigation.

Real-time RT-PCR of tyrosine hydroxylase to detect bone marrow involvement in advanced neuroblastoma.

Neuroblastoma (NB) is characterised by the secretion of catecholamines in approximately 95% of patients. Tyrosine hydroxylase is the rate-limiting enzyme in catecholamine biosynthesis pathway. Expression of the tyrosine hydroxylase gene (TH) is regulated in a tissue-specific manner during neonatal development and differentiation, therefore TH mRNA expression is a specific tumour marker for NB. Here we present a real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay using TaqMan technology for detection and quantification of TH mRNA in bone marrow (BM) NB patients. The degree of TH expression was derived from the ratio of the mRNAs of this gene and the reference gene, beta-actin. A ratio greater than 3x10(-2) was considered as positive for TH mRNA presence. Samples were also examined for TH mRNA by first and nested RT-PCR. Seventeen BM samples from 4 patients with disseminated NB (3 stage IV and 1 stage IVs) were evaluated at diagnosis and during treatment. We found a variable degree of TH expression ranging from 0.0344 to 26.3370 in 12/17 positive samples, while no TH mRNA (value lower than 3x10(-2)) was detected in 5/17 samples obtained after consolidation therapy. Our results show a moderate concordance between different qualitative RT-PCR methods and real-time RT-PCR. The real-time RT-PCR results seem to fit better with the natural short-term clinical follow-up of the evaluated patients, with respect to qualitative methods. Real-time TH RT-PCR could therefore be of clinical value for the assessment of a patient's prognosis by monitoring minimal residual disease (MRD).