Upregulation of soluble and membrane-bound human leukocyte antigen G expression is primarily observed in the milder histopathological stages of chronic hepatitis C virus infection

Chronic hepatitis C virus (HCV) infection is a worldwide health problem that may evolve to cirrhosis and hepatocellular carcinoma. Incompletely understood immune system mechanisms have been associated with impaired viral clearance. The nonclassical class I human leukocyte antigen G (HLA-G) molecule may downregulate immune system cell functions exhibiting well-recognized tolerogenic properties. HCV genotype was analyzed in chronic HCV-infected patients. Because HLA-G expression may be induced by certain viruses, we evaluated the presence of HLA-G in the liver microenvironment obtained from 89 biopsies of patients harboring chronic HCV infection and stratified according to clinical and histopathological features. Overall, data indicated that HCV genotype 1 was predominant, especially subgenotype 1a, with a prevalence of 87%. HLA-G expression was observed in 45(51%) liver specimens, and it was more frequent in milder stages of chronic hepatitis (67.4%) than in moderate (27.8%; p = 0.009) and severe (36.0%; p = 0.021) stages of the disease. Altogether, these results suggest that the expression of HLA-G in the context of HCV is a complex process modulated by many factors, which may contribute to an immunologic environment favoring viral persistence. However, because the milder forms predominantly expressed HLA-G, a protective role of this molecule may not be excluded. (C) 2012 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.

Prevalence of non-organ-specific autoantibodies in a rural community from northeastern Brazil: a population-based study

Non-organ-specific autoantibodies (NOSA) are well-recognized diagnostic markers of autoimmune hepatitis (All-l) and primary biliary cirrhosis (PBC), but can also be observed in patients with viral hepatitis as well as in healthy subjects. The aim of this study was to evaluate the prevalence of NOSA in subjects living in a rural community in Brazil and to correlate their occurrence with the presence of liver disease. Seven hundred twenty-five apparently healthy subjects were randomly selected for assessment of antinuclear (ANA), anti-smooth muscle (SMA), antimitochondrial (AMA), anti-liver/kidney microsome type 1, and anti-liver cytosol type 1 antibodies. Subjects with those NOSA were evaluated for the presence of AIH, PBC, and viral hepatitis. Reactivities for all NOSA, SMA, ANA, and AMA were detected, respectively, in 14, 10, 4, and 0.1% of subjects, with a mean titer of 1:40. NOSA-positive subjects were significantly older and more frequently females. No correlation was observed between the occurrence of NOSA and PBC. AIH, or viral hepatitis. The prevalence of NOSA in Brazilians was 14%. They were usually low titer. NOSA were more frequently observed in females and older subjects and their presence was not correlated with the presence of AIH, PBC, or viral hepatitis. (C) 2012 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.

Organ-specific autoantibodies and autoimmune diseases in juvenile systemic lupus erythematosus and juvenile dermatomyositis patients

Objectives To our knowledge, no study assessed simultaneously a variety of organ-specific autoantibodies and the prevalence of organ-specific autoimmune diseases in juvenile systemic lupus erythematosus (ISLE) and juvenile dermatomyositis (JDM). Therefore, the purpose of this study was to evaluate organ-specific autoantibodies and autoimmune diseases in JSLE and JDM patients. Methods Forty-one JSLE and 41 JDM patients were investigated for autoantibodies associated with autoimmune hepatitis, primary biliary cirrhosis, type I diabetes mellitus (TIDM, autoimmune thyroiditis (AT), autoimmune gastritis and coeliac disease (CD). Patients with positive antibodies were investigated for the respective organ-specific autoimmune diseases. Results Mean age at diagnosis was higher in ISLE compared to JDM patients (10.3 +/- 3.4 vs. 7.3 +/- 3.1 years, p=0.0001). The frequencies of organ-specific autoantibodies were similar in JSLE and JDM patients (p>0.05). Of note, a high prevalence of TIDM and AT autoantibodies was observed in both groups (20% vs. 15%, p=0.77 and 24% vs. 15%, p=0.41; respectively). Higher frequencies of ANA (93% vs. 59%, p=0.0006), anti-dsDNA (61% vs. 2%, p<0.0001), anti-Ro, anti-Sm, anti-RNP, anti-La and IgG-aCL were observed in JSLE (p<0.05). Organ-specific autoimmune diseases were evidenced only in ISLE patients (24% vs. 0%, p=0.13). Two ISLE patients had TIDM associated with Hashimoto thyroiditis and another had subclinical thyroiditis. Another JSLE patient had CD diagnosis based on iron deficiency anaemia, anti-endomysial antibody, duodenal biopsy compatible to CD and response to a gluten-free diet. Conclusions Organ-specific diseases were observed solely in ISLE patients and required specific therapy. The presence of these antibodies recommends the evaluation of organ-specific diseases and a rigorous follow-up.

Coffee and Caffeine Protect against Liver Injury Induced by Thioacetamide in Male Wistar Rats

Coffee intake has been inversely related to the incidence of liver diseases, although there are controversies on whether these beneficial effects on human health are because of caffeine or other specific components in this popular beverage. Thus, this study evaluated the protective effects of coffee or caffeine intake on liver injury induced by repeated thioacetamide (TAA) administration in male Wistar rats. Rats were randomized into five groups: one untreated group (G1) and four groups (G2G5) treated with the hepatotoxicant TAA (200 similar to mg/kg b.w., i.p.) twice a week for 8 similar to weeks. Concomitantly, rats received tap water (G1 and G2), conventional coffee (G3), decaffeinated coffee (G4) or 0.1% caffeine (G5). After 8 similar to weeks of treatment, rats were killed and blood and liver samples were collected. Conventional and decaffeinated coffee and caffeine intake significantly reduced serum levels of alanine aminotransferase (ALT) (p similar to<similar to 0.001) and oxidized glutathione (p similar to<similar to 0.05), fibrosis/inflammation scores (p similar to<similar to 0.001), collagen volume fraction (p similar to<similar to 0.01) and transforming growth factor beta-1 (TGF-beta 1) protein expression (p similar to=similar to 0.001) in the liver from TAA-treated groups. In addition, conventional coffee and caffeine intake significantly reduced proliferating cellular nuclear antigen (PCNA) S-phase indexes (p similar to<similar to 0.001), but only conventional coffee reduced cleaved caspase-3 indexes (p similar to<similar to 0.001), active metalloproteinase 2 (p similar to=similar to 0.004) and the number of glutathione S-transferase placental form (GST-P)-positive preneoplastic lesions (p similar to<similar to 0.05) in the liver from TAA-treated groups. In conclusion, conventional coffee and 0.1% caffeine intake presented better beneficial effects than decaffeinated coffee against liver injury induced by TAA in male Wistar rats.

Plasmatic higher levels of homocysteine in Non-alcoholic fatty liver disease (NAFLD)

Background Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease, which includes a spectrum of hepatic pathology such as simple steatosis, steatohepatitis, fibrosis and cirrhosis. The increased serum levels of homocysteine (Hcy) may be associated with hepatic fat accumulation. Genetic mutations in the folate route may only mildly impair Hcy metabolism. The aim of this study was to investigate the relation between liver steatosis with plasma homocysteine level and MTHFR C677T and A1298C polymorphisms in Brazilian patients with NAFLD. Methods Thirty-five patients diagnosed with NAFLD by liver biopsy and forty-five healthy controls neither age nor sex matched were genotyped for C677T and A1298C MTHFR polymorphisms using PCR-RFLP and PCR-ASA, respectively, and Hcy was determined by HPLC. All patients were negative for markers of Wilson’s, hemochromatosis and autoimmune diseases. Their daily alcohol intake was less than 100 g/week. A set of metabolic and serum lipid markers were also measured at the time of liver biopsies. Results The plasma Hcy level was higher in NAFLD patients compared to the control group (p = 0.0341). No statistical difference for genotypes 677C/T (p = 0.110) and 1298A/C (p = 0.343) in patients with NAFLD and control subjects was observed. The genotypes distribution was in Hardy-Weinberg equilibrium (677C/T p = 0.694 and 1298 A/C p = 0.188). The group of patients and controls showed a statistically significant difference (p < 0.001) for BMI and HOMA_IR, similarly to HDL cholesterol levels (p < 0,006), AST, ALT, γGT, AP and triglycerides levels (p < 0.001). A negative correlation was observed between levels of vitamin B12 and Hcy concentration (p = 0.005). Conclusion Our results indicate that plasma Hcy was higher in NAFLD than controls. The MTHFR C677T and A1298C polymorphisms did not differ significantly between groups, despite the 677TT homozygous frequency was higher in patients (17.14%) than in controls (677TT = 4.44%) (p > 0.05). The suggested genetic susceptibility to the MTHFR C677T and A1298C should be confirmed in large population based studies.

Hérnias abdominais e inguinais em pacientes cirróticos: qual é a melhor conduta?

INTRODUÇÃO: Tradicionalmente os procedimentos abdominais eletivos em pacientes cirróticos têm sido amplamente desencorajados graças à elevada morbi-mortalidade consequente às complicações da cirrose, descritas por diversos autores. Outros serviços, em contrapartida, obtiveram resultados distintos, advogando a favor de cirurgia eletiva. MÉTODOS: Uma revisão de artigos utilizando-se a palavras "abdominal wall hernia" e "cirrhotic patients" foi realizada na base de dados PubMed. Dos resultados obtidos, 28 artigos foram considerados para elaboração desta revisão. RESULTADOS: Pôde-se observar que a incidência de hérnias em parede abdominal é relativamente elevada em pacientes cirróticos, sendo que muitas delas têm evolução desfavorável e requerem tratamento cirúrgico específico. Com o advento do sistema de alocação de órgãos baseados no escore de MELD, muitos centros estão repensando suas condutas em situações como esta, dado que muitos dos pacientes em questão encontram-se em lista de espera para transplante hepático. Dessa forma a cirurgia eletiva tem conquistado maior papel no manejo desta condição com intuito de diminuir morbi-mortalidade nesses pacientes. Além disso, a qualidade de vida mostrou-se um importante fator a ser considerado, estando muito prejudicada nesta condição. CONCLUSÃO: Poucos estudos com grandes amostragens foram conduzidos até o momento e não há consenso sobre qual conduta é a mais indicada levando em consideração taxas de morbi-mortalidade.

Case for diagnosis

Infantile digital fibromatosis or Reye's tumor is a benign fibroproliferative tumor, the etiopathogenesis of which has yet to be fully clarified. It typically presents at birth or in the first year of life and is characterized by a firm, flesh colored or erythematous nodule or nodules located on the digits. These lesions tend to regress spontaneously.

Resultados da ressecção de hemangiomas infantis nasais em fase proliferativa: abordagem segura para os tumores centrais da face

INTRODUÇÃO: O hemangioma infantil é o tumor benigno mais comum da infância, predominando na região cervicofacial. É caracterizado por apresentar 3 fases distintas, observando-se frequentemente regressão espontânea dessas lesões. No entanto, sequelas residuais ou deformidades das estruturas anatômicas em crescimento podem ocorrer. A abordagem cirúrgica precoce e definitiva é indicada, em decorrência da localização dos hemangiomas nasais e seu potencial desfigurante, visando à obtenção de bons resultados estéticos e preservação anatômica. O objetivo do presente estudo foi analisar os resultados da abordagem cirúrgica definitiva para hemangiomas proliferativos nasais, com base em uma avaliação objetiva. MÉTODO: No período de 1997 a 2009, 20 pacientes portadores de hemangiomas nasais em fase proliferativa foram submetidos a tratamento cirúrgico. As lesões foram avaliadas segundo local de acometimento e tratamento realizado. Foram analisados índices de complicações e necessidade de procedimentos adicionais. Os resultados estéticos foram avaliados por avaliadores independentes. RESULTADOS: As lesões estavam localizadas na ponta nasal em 50% dos pacientes; no dorso, em 20%; em todas as subunidades, em 15%; nas áreas paranasais, em 10%; e na unidade alar, em 5%. A ressecção foi total em 60% dos pacientes e subtotal em 40%. O período médio de acompanhamento foi de 42,6 meses. A média de procedimentos cirúrgicos por paciente foi de 1,3 + 0,7. Nenhuma complicação importante foi observada. Os resultados foram positivamente avaliados quanto a redução do volume da lesão e melhora do contorno facial, corroborando a conduta proposta. CONCLUSÕES: No manejo dos hemangiomas nasais, o tratamento cirúrgico definitivo pode ser considerado uma alternativa segura e eficaz, com baixas taxas de complicação.

Niemann-Pick disease type C symptomatology: an expert-based clinical description

Niemann-Pick disease type C (NP-C) is a rare, progressive, irreversible disease leading to disabling neurological manifestations and premature death. The estimated disease incidence is 1:120,000 live births, but this likely represents an underestimate, as the disease may be under-diagnosed due to its highly heterogeneous presentation. NP-C is characterised by visceral, neurological and psychiatric manifestations that are not specific to the disease and that can be found in other conditions. The aim of this review is to provide non-specialists with an expert-based, detailed description of NP-C signs and symptoms, including how they present in patients and how they can be assessed. Early disease detection should rely on seeking a combination of signs and symptoms, rather than isolated findings. Examples of combinations which are strongly suggestive of NP-C include: splenomegaly and vertical supranuclear gaze palsy (VSGP); splenomegaly and clumsiness; splenomegaly and schizophrenia-like psychosis; psychotic symptoms and cognitive decline; and ataxia with dystonia, dysarthria/dysphagia and cognitive decline. VSGP is a hallmark of NP-C and becomes highly specific of the disease when it occurs in combination with other manifestations (e.g. splenomegaly, ataxia). In young infants (<2 years), abnormal saccades may first manifest as slowing and shortening of upward saccades, long before gaze palsy onset. While visceral manifestations tend to predominate during the perinatal and infantile period (2 months–6 years of age), neurological and psychiatric involvement is more prominent during the juvenile/adult period (>6 years of age). Psychosis in NP-C is atypical and variably responsive to treatment. Progressive cognitive decline, which always occurs in patients with NP-C, manifests as memory and executive impairment in juvenile/adult patients. Disease prognosis mainly correlates with the age at onset of the neurological signs, with early-onset forms progressing faster. Therefore, a detailed and descriptive picture of NP-C signs and symptoms may help improve disease detection and early diagnosis, so that therapy with miglustat (Zavesca®), the only available treatment approved to date, can be started as soon as neurological symptoms appear, in order to slow disease progression.

Trasmissione post-natale del citomegalovirus attraverso il latte materno al neonato VLBW

Introduction Postnatal human cytomegalovirus (CMV) infection is usually asymptomatic in term babies, while preterm infants are more susceptible to symptomatic CMV infection. Breastfeeding plays a dominant role in the epidemiology of transmission of postnatal CMV infection, but the risk factors of symptomatic CMV infection in preterm infants are unknown. Patients and Methods Between December 2003 and August 2006, eighty Very Low Birth Weight (VLBW) preterm infants (gestational age ≤ 32 weeks and birth weight < 1500 g), admitted to the Neonatal Intensive Care Unit of St Orsola-Malpighi General Hospital, Bologna were recruited. All of them were breastfed for at least one month. During the first week of life, serological test for CMV was performed on maternal blood. Furthermore, urinary CMV culture was performed in all the infants in order to exclude a congenital CMV infection. Urine samples from each infant were collected and processed for CMV culture once a week. Once every 15 days a blood sample was taken from each infant to evaluate the complete blood count, the hepatic function and the C reactive protein. In addition, samples of fresh breast milk were processed weekly for CMV culture. A genetic analysis of virus variant was performed in the urine of the infected infants and in their mother’s milk to confirm the origin of infection. Results We evaluated 80 VLBW infants and their 68 mothers. Fifty-three mothers (78%) were positive for CMV IgG antibodies, and 15 (22%) were seronegative. In the seronegative group, CMV was never isolated in breast milk, and none of the 18 infants developed viruria; in the seropositive group, CMV was isolated in 21 out of 53 (40%) mother’s milk. CMV was detected in the urine samples of 9 out of 26 (35%) preterm infants, who were born from 21 virolactia positive mothers. Six of these infants had clinically asymptomatic CMV infection, while 3 showed a sepsis-like illness with bradycardia, tachypnea and repeated desaturations. Eight out of nine infants showed abnormal hematologic values. The detection of neutropenia was strictly related to CMV infection (8/9 infected infants vs 17/53 non infected infants, P<.005), such as the detection of an increase in conjugated bilirubin (3/9 infected infants vs 2/53 non infected infants, P<.05). The degree of neutropenia was not different between the two groups (infected/non infected). The use of hemoderivatives (plasma and/or IgM–enriched immunoglobulin) in order to treat a suspected/certain infection in newborn with GE< 28 ws was seen as protective against CMV infection (1/4 infected infants vs 18/20 non infected infants [GE<28 ws]; P<.05). Furthermore, bronchopulmonary dysplasia (defined both as oxygen-dependency at 30 days of life and 36 ws of postmenstrual age) correlated with symptomatic infection (3/3 symptomatic vs 0/6 asymptomatic: P<.05). Conclusion Our data suggest that CMV infection transmitted to preterm newborn through human milk is always asymptomatic when newborns are clinically stable. Otherwise, the infection can worsen a preexisting disease such as bronchopulmonary dysplasia. Human milk offers many nutritional and psychological advantages to preterm newborns: according to our data, there’s no reason to contraindicate it neither to pasteurize the milk of all the mothers of preterm infants who are CMV seropositive.

Il ruolo dell'ecografia prenatale nell'infezione congenita da Citomegalovirus

OBJECTIVE: To assess the effectiveness of ultrasound in the antenatal prediction of symptomatic congenital cytomegalovirus infection. STUDY DESIGN: The sonograms of 650 fetuses from mothers with primary cytomegalovirus infection were correlated to fetal/neonatal outcome. Infection status was disclosed by viral urine isolation at birth or CMV tissue inclusions at autopsy. Classification of symptomatic disease was based on postnatal clinical/laboratory findings or macroscopic evidence of tissue damage at autopsy. RESULTS: Ultrasound abnormalities were found in 51/600 (8.5%) mothers with primary infection and in 23/154 congenitally infected fetuses (14.9%). Symptomatic congenital infection resulted in 18/23 and 68/131 cases with or without abnormal sonographic findings, respectively. Positive predictive values of ultrasound versus symptomatic congenital infection was 35.3% relating to all fetuses/infants from mothers with primary infection and 78.3% relating to fetuses/infants with congenital infection. CONCLUSION: When fetal infection status is unknown, ultrasound abnormalities only predict symptomatic congenital infection in a third of cases.

Le cardiomiopatie nell'infanzia: quadri eziologici ed evolutivi nei primi anni di vita

Premessa: nell’aprile 2006, l’American Heart Association ha approvato la nuova definizione e classificazione delle cardiomiopatie (B. J. Maron e coll. 2006), riconoscendole come un eterogeneo gruppo di malattie associate a disfunzione meccanica e/o elettrica riconducibili ad un ampia variabilità di cause. La distinzione tra le varie forme si basa non più sui processi etiopatogenetici che ne sono alla base, ma sulla modalità di presentazione clinica della malattia. Si distinguono così le forme primarie, a prevalente od esclusivo interessamento cardiaco, dalle forme secondarie in cui la cardiomiopatia rientra nell’ambito di un disordine sistemico dove sono evidenziabili anche disturbi extracardiaci. La nostra attenzione è, nel presente studio, focalizzata sull’analisi delle cardiomiopatie diagnosticate nei primi anni di vita in cui si registra una più alta incidenza di forme secondarie rispetto all’adulto, riservando un particolare riguardo verso quelle forme associate a disordini metabolici. Nello specifico, il nostro obiettivo è quello di sottolineare l’influenza di una diagnosi precoce sull’evoluzione della malattia. Materiali e metodi: abbiamo eseguito uno studio descrittivo in base ad un’analisi retrospettiva di tutti i pazienti giunti all’osservazione del Centro di Cardiologia e Cardiochirurgia Pediatrica e dell’ Età Evolutiva del Policlinico S. Orsola- Malpighi di Bologna, dal 1990 al 2006, con diagnosi di cardiomiopatia riscontrata nei primi due anni di vita. Complessivamente sono stati studiati 40 pazienti di cui 20 con cardiomiopatia ipertrofica, 18 con cardiomiopatia dilatativa e 2 con cardiomiopatia restrittiva con un’età media alla diagnosi di 4,5 mesi (range:0-24 mesi). Per i pazienti descritti a partire dal 2002, 23 in totale, sono state eseguite le seguenti indagini metaboliche: emogasanalisi, dosaggio della carnitina, metabolismo degli acidi grassi liberi (pre e post pasto), aminoacidemia quantitativa (pre e post pasto), acidi organici, mucopolisaccaridi ed oligosaccaridi urinari, acilcarnitine. Gli stessi pazienti sono stati inoltre sottoposti a prelievo bioptico di muscolo scheletrico per l’analisi ultrastrutturale, e per l’analisi dell’attività enzimatica della catena respiratoria mitocondriale. Nella stessa seduta veniva effettuata la biopsia cutanea per l’eventuale valutazione di deficit enzimatici nei fibroblasti. Risultati: l’età media alla diagnosi era di 132 giorni (range: 0-540 giorni) per le cardiomiopatie ipertrofiche, 90 giorni per le dilatative (range: 0-210 giorni) mentre le 2 bambine con cardiomiopatia restrittiva avevano 18 e 24 mesi al momento della diagnosi. Le indagini metaboliche eseguite sui 23 pazienti ci hanno permesso di individuare 5 bambini con malattia metabolica (di cui 2 deficit severi della catena respiratoria mitocondriale, 1 con insufficienza della β- ossidazione per alterazione delle acilcarnitine , 1 con sindrome di Barth e 1 con malattia di Pompe) e un caso di cardiomiopatia dilatativa associata a rachitismo carenziale. Di questi, 4 sono deceduti e uno è stato perduto al follow-up mentre la forma associata a rachitismo ha mostrato un netto miglioramento della funzionalità cardiaca dopo appropriata terapia con vitamina D e calcio. In tutti la malattia era stata diagnosticata entro l’anno di vita. Ciò concorda con gli studi documentati in letteratura che associano le malattie metaboliche ad un esordio precoce e ad una prognosi infausta. Da un punto di vista morfologico, un’evoluzione severa si associava alla forma dilatativa, ed in particolare a quella con aspetto non compaction del ventricolo sinistro, rispetto alla ipertrofica e, tra le ipertrofiche, alle forme con ostruzione all’efflusso ventricolare. Conclusioni: in accordo con quanto riscontrato in letteratura, abbiamo visto come le cardiomiopatie associate a forme secondarie, ed in particolare a disordini metabolici, sono di più frequente riscontro nella prima infanzia rispetto alle età successive e, per questo, l’esordio molto precoce di una cardiomiopatia deve essere sempre sospettata come l’espressione di una malattia sistemica. Abbiamo osservato, inoltre, una stretta correlazione tra l’età del bambino alla diagnosi e l’evoluzione della cardiomiopatia, registrando un peggioramento della prognosi in funzione della precocità della manifestazione clinica. In particolare la diagnosi eseguita in epoca prenatale si associava, nella maggior parte dei casi, ad un’evoluzione severa, comportandosi come una variabile indipendente da altri fattori prognostici. Riteniamo, quindi, opportuno sottoporre tutti i bambini con diagnosi di cardiomiopatia effettuata nei primi anni di vita ad uno screening metabolico completo volto ad individuare quelle forme per le quali sia possibile intraprendere una terapia specifica o, al contrario, escludere disordini che possano controindicare, o meno, l’esecuzione di un trapianto cardiaco qualora se ne presenti la necessità clinica.

Significato prognostico dell'ecografia cerebrale nei neonati con infezione congenita da citomegalovirus

Background: Congenital cytomegalovirus (CMV) infection may lead to cerebral injury and neurodevelopmental delay. Cranial computed tomography (CT) is currently the standard imaging technique for predicting the outcome of CMV infected patients. Ultrasound (US) is a safe means to assess the extent of cerebral injury due to CMV infection in neonates, and unlike CT, is readily available at the bedside. Aim: To report the accuracy of US in predicting neurodevelopmental and sensorineural outcome in patients with congenital CMV infection. Study design: 57 newborns with congenital CMV infection underwent brain US and were followed prospectively for motor skills, developmental quotient and hearing function. Results: An abnormal US was found in 12/57 newborns. At least one sequela (Developmental Quotient < 85, motor delay, sensorineural hearing loss) was present in 10/11 surviving children with abnormal US (1 patient died in the neonatal period) vs 3/45 newborns with normal US (OR for death or poor outcome: 154, CI 17.3-1219.6, p<0.001, positive predictive value 91.7%, negative predictive value 93.3%). Conclusion: A good correlation is shown between ultrasound abnormalities and the prediction of outcome, suggesting that US may be used to study and follow CMV infected neonates. Our findings await confirmation in a larger population.

Mass spectrometry-based protein profiling strategies for biomarker discovery in liver and inflammatory bowel diseases

The study of protein expression profiles for biomarker discovery in serum and in mammalian cell populations needs the continuous improvement and combination of proteins/peptides separation techniques, mass spectrometry, statistical and bioinformatic approaches. In this thesis work two different mass spectrometry-based protein profiling strategies have been developed and applied to liver and inflammatory bowel diseases (IBDs) for the discovery of new biomarkers. The first of them, based on bulk solid-phase extraction combined with matrix-assisted laser desorption/ionization - Time of Flight mass spectrometry (MALDI-TOF MS) and chemometric analysis of serum samples, was applied to the study of serum protein expression profiles both in IBDs (Crohn’s disease and ulcerative colitis) and in liver diseases (cirrhosis, hepatocellular carcinoma, viral hepatitis). The approach allowed the enrichment of serum proteins/peptides due to the high interaction surface between analytes and solid phase and the high recovery due to the elution step performed directly on the MALDI-target plate. Furthermore the use of chemometric algorithm for the selection of the variables with higher discriminant power permitted to evaluate patterns of 20-30 proteins involved in the differentiation and classification of serum samples from healthy donors and diseased patients. These proteins profiles permit to discriminate among the pathologies with an optimum classification and prediction abilities. In particular in the study of inflammatory bowel diseases, after the analysis using C18 of 129 serum samples from healthy donors and Crohn’s disease, ulcerative colitis and inflammatory controls patients, a 90.7% of classification ability and a 72.9% prediction ability were obtained. In the study of liver diseases (hepatocellular carcinoma, viral hepatitis and cirrhosis) a 80.6% of prediction ability was achieved using IDA-Cu(II) as extraction procedure. The identification of the selected proteins by MALDITOF/ TOF MS analysis or by their selective enrichment followed by enzymatic digestion and MS/MS analysis may give useful information in order to identify new biomarkers involved in the diseases. The second mass spectrometry-based protein profiling strategy developed was based on a label-free liquid chromatography electrospray ionization quadrupole - time of flight differential analysis approach (LC ESI-QTOF MS), combined with targeted MS/MS analysis of only identified differences. The strategy was used for biomarker discovery in IBDs, and in particular of Crohn’s disease. The enriched serum peptidome and the subcellular fractions of intestinal epithelial cells (IECs) from healthy donors and Crohn’s disease patients were analysed. The combining of the low molecular weight serum proteins enrichment step and the LCMS approach allowed to evaluate a pattern of peptides derived from specific exoprotease activity in the coagulation and complement activation pathways. Among these peptides, particularly interesting was the discovery of clusters of peptides from fibrinopeptide A, Apolipoprotein E and A4, and complement C3 and C4. Further studies need to be performed to evaluate the specificity of these clusters and validate the results, in order to develop a rapid serum diagnostic test. The analysis by label-free LC ESI-QTOF MS differential analysis of the subcellular fractions of IECs from Crohn’s disease patients and healthy donors permitted to find many proteins that could be involved in the inflammation process. Among them heat shock protein 70, tryptase alpha-1 precursor and proteins whose upregulation can be explained by the increased activity of IECs in Crohn’s disease were identified. Follow-up studies for the validation of the results and the in-depth investigation of the inflammation pathways involved in the disease will be performed. Both the developed mass spectrometry-based protein profiling strategies have been proved to be useful tools for the discovery of disease biomarkers that need to be validated in further studies.

Memoria e rappresentazione della violenza storica nella letteratura del secondo Novecento

Partendo dal problema del rapporto fra la ricostruzione di un evento storico e lo statuto del testo che lo ricostruisce, la tesi si concentra nella lettura di opere riguardanti la Seconda guerra mondiale. Sono in questo senso cruciali due opere autobiografiche trattate nella prima parte del lavoro, Rue Labat Rue Ordener di Sarah Kofman (1993) e Kindheitsmuster di Christa Wolf (1976). In questi due testi la dottoranda prova a recuperare da una parte la rimemorazione letteraria di due esperienze infantili della guerra insieme opposte e complementari dal punto di vista del posizionamento della testimonianza. Il testo della Wolf ibrida la narrazione finzionale e la memoria dell’evento storico vissuto nel ricordo di una bambina tedesca, il testo della Kofman recupera in una maniera quasi psicanalitica la memoria di una bambina ebrea vittima inconsapevole e quesi incosciente della Shoah. Di fronte a due topoi apparentemente già piu volte ripercorsi nella letteratura critica del trauma postconflitto, la tesi in questione cerca di individuare il costituirsi quasi inevitabile di un soggetto identitario che osserva, vive e successivamente recupera e racconta il trauma. Se alcune posizioni della moderna storiografia e della contemporanea riflessione sulla scrittura storiografica, sottolineano la forza e l’importanza dell’elemento narrativo all’interno della ricostruzione storica e dell’analisi dei documenti, questa tesi sembra indicare una via possibile di studio per la letteratura comparata. Una via, cioè, che non si soffermi sui fattori e sui criteri veridizionali del testo, criteri e fattori che devono restare oggetto di studio per gli storici , ma che piuttosto indaghi sul nesso ineludibile e fondativo che la scrittura stessa svela e pone in essere: il trauma, l’irrompere dell’evento storico nell’individuo diventa elemento costitutivo della propria identità, elemento al quale è difficile dare una posizione stabile ma che allo stesso tempo non si può evitare di raccontare, di mettere in discorso. Nella narrazione letteraria di eventi storici esiste dunque un surplus di senso che sta tutto nella costituzione di una posizione dalla quale raccontare, di un punto di vista. Il punto di vista (come ci ricorda De Certeau ne Les lieux des autres in quel saggio dedicato ai cannibali di Montaigne,che viene poi ripreso senza essere mai citato da Ginzburg ne Il filo e le tracce) non è mai dato a priori nel discorso, è il risultato di un conflitto e di una lotta. Il testo che rende conto e recupera la memoria di un passato storico, in particolare di un passato storico conflittuale, di una guerra, di una violenza, per quanto presenti un punto di vista preciso e posizionato, per quanto possa apparire un frutto di determinate strategie testuali e di determinati obiettivi pragmatici, è pur sempre una narrazione il cui soggetto porta in sé, identitariamente, le ferite e i traumi dell’evento storico. Nei casi di Wolf e Kofman abbiamo quindi un rispecchiamento reciproco che fra il tentativo di una ricostruzione della memoria infantile e il recupero dell’elemento intersoggettivo e storico si apre alla scrittura e alla narrazione. La posizione del soggetto che ha vissuto l’irrompere del dramma storico nel discorso lo costituisce e lo delega a essere colui che parla e colui che vede. In un qualche modo la Storia per quanto possa essere creatrice di eventi e per quanto possa trasformare l’esistenza del soggetto non è essa stessa percepibile finché non si posiziona attraverso il soggetto trasformato e modificato all’interno del discorso. In questa continua ricerca di un equilibrio possibile fra realtà e discorso si pone il problema dell’essere soggetto in mezzo ad altri soggetti. E questo in un duplice aspetto: nell’aspetto della rappresentazione dell’altro, cioè nel problema di come la memoria riorganizzi e ricrei i soggetti in gioco nell’evento storico; e poi nella rappresentazione di se stesso per gli altri, nella rappresentazione cioe del punto di vista. Se nel romanzo autobiografico di Kofman tutta la storia veniva a ricondursi alla narrazione privata del soggetto che, come in una seduta psicanalitica recupera e insieme si libera del proprio conflitto interiore, della propria memoria offesa; se nel romanzo di Wolf si cercava un equilibrio fra una soggettivita infantile ormai distante in terza persona e una soggettività rammemorante che prendeva posizione nel romanzo nella seconda persona; la cerniera sia epistemologica sia narratologica fra la prima e la seconda parte della tesi pare essere Elsa Morante e il suo romanzo La Storia. L’opera della Morante sembra infatti farsi pieno carico della responsabilità di non poter piu ridurre la narrazione del trauma alla semplice presa in carico del soggetto autobiografico. Il soggetto che, per dirla ancora con De Certeau, può esprimere il proprio punto di vista perche in qualche modo si è salvato dalla temperie della storia, non si pone nel discorso come punto di inizio e di fine di qualsiasi percezione del trauma, ma si incarna in uno o più personaggi che in un qualche modo rappresentino l’irrapresentabile e l’irrapresentato. La storia diventa quindi elemento non costitutivo di un'identità capace di ri-raccontarsi o almeno non solo, diventa fattore costitutivo di un’identità capace di raccontare l’altro, anzi gli altri, tutti coloro che il conflitto, la violenza ha in un qualche modo cancellato. Così accade all’infanzia tradita e offesa del piccolo Useppe, che viene soffocato non solo nella sua possibilita di svilupparsi, di essere punto di vista del discorso, ma anche nella possibilita di essere osservatore vivo dell’evento; cosi accade a Ida, donna e madre, che la Storia lentamente e inesorabilmente spersonalizza riducendola a essere soggetto passivo e vittima degli eventi. Ecco quindi che la strada aperta dalla Morante permette alla memoria di proiettarsi in una narrazione comune e di condividere e suddividere la posizione centrale del soggetto in una costellazione differente di soggetti. A questo punto si apre, attraverso le tecniche della storia orale e la loro narrativizzazione, una strategia di recupero della memoria evidenziata nell’ultima parte della tesi. La strada intrapresa da autori come i Wu Ming e come Andrea Levy ne è un esempio. Sganciati per evidenti ragioni biografiche e anagrafiche (sono tutti nati ben dopo la fine del secondo conflitto mondiale) da qualsiasi tentazione autobiografica, i primi intraprendono una vera e propria ridistribuzione dei punti di vista sulla storia. In romanzi come Manituana si viene a perdere, almeno a un primo e forse piu superficiale livello, qualsiasi imposizione fissa del punto di vista. Una molteplicità di soggetti si prende carico di raccontare la storia da differenti posizioni, ma la apparente molteplicità degli sguardi non si riduce a una frammentazione dell’etica del racconto quanto piuttosto alla volontà di fare emergere tra gli altri anche il punto di vista dello sconfitto e dell’inerme. Al passato oscuro della violenza storica si contrappone in un qualche modo la messa in discorso del soggetto che cerca attraverso la costituzione non solo di un soggetto ma di una pluralitàdi voci , di ritrovare un’ armonia, di riassimilare la propria memoria condividendola nel testo letterario.