Contribution to the study of diseases in Italy caused by Coxsackie B group of viruses I. Clinical and virological aspects of an outbreak of Bornholm disease

Summary The virological findings obtained during an outbreak of Bornholm disease which took place in the summer 1957 in Cermignano, in the province of Teramo, Italy, are described. During this study it was possible to isolate 15 strains of a Goxsackie B2 type virus. 3 strains were isolated simultaneously from rectal and pharyn geal swabs, while 9 strains were isolated from pharyngeal swabs only.     

Attentional, emotional and hormonal data in subjects of different ages

The differences in attentional style among subjects of different ages and the influence of emotionality on the attentional components were studied for a limited experimental period. Variation in the hormonal data and its relation to behavioural parameters were also evaluated. The subjects enrolled in the study were divided into four age groups (A 18–29, B 30–45, C 46–59, D 60–77 years). The attentional tests involved different types of attention: alert, go/no-go, divided attention and working memory. Emotionality was assessed on the basis of skin conductance, heart rate and frontalis muscle tone. Testosterone (T), free testosterone (fT), non-specifically bound testosterone (NST), sex hormone binding globulin (sHBG), oestradiol, cortisol and adrenocorticotrophic hormone were determined in the plasma. The data were analysed to identify endocrine and behavioural differences related to sex and age. The results showed an influence of age on reaction time (RT) and RT variability. This was particularly evident for groups C and D with respect to A in the simple (alert) and complex RT tests (go/no-go and working memory). Divided attention, with the highest RT, showed a clear distinction between group A and the other groups. The difference in frontalis electromyography (EMG) (test vs control) increased with age, while the autonomic responses (skin conductance and heart rate) did not vary. In most attentional tests, the age-related reduction of RT was associated with increased T, fT and NST and decreased cortisol.     

Peripheral cytokine release in Alzheimer patients: correlation with disease severity

Various studies suggested that inflammation is involved in the pathogenesis of Alzheimer's disease (AD). We investigated cytokine release from LPS-stimulated blood cells of 32 AD patients, with different disease severity, compared to 16 age-related controls. A significant decrease of IL-1beta and IL-6 secretion was observed in severely demented patients; TNF-alpha release was also decreased, but not significantly. By contrast, mild and moderate patients showed a cytokine release similar to controls. IL-1beta, IL-6 and TNF-alpha secretion was negatively correlated with the severity of dementia, quantified by the MMSE. Our data suggest that alterations of the immune profile are associated with AD progression.     

Antigenic and immunogenic mimicry of the HER2/neu oncoprotein by phage-displayed peptides

To recover peptides that antigenically and immunogenically mimic the p185^HER2 oncoprotein, we selected the phage-peptide libraries pVIII-9aa and pVIII-9aa. Cys using murine monoclonal antibodies (mAb) MGr2 and MGr6, directed against two distinct epitopes of the p185^HER2 extracellular domain. Phagedisplayed peptides containing consensus amino acid motifs were recovered and shown to compete specifically for mAb binding on tumor cells that overexpress p185^HER2. The deduced amino acid sequence of the peptides suggests that both epitopes defined by the mAb on p185^HER2 are discontinuous and that hydrophobic interactions are involved in binding with the mAb. A phage clone displaying the GPLDSLFAQ peptide elicited a specific immune response against the p185^HER2 in BALB/c mice, demonstrating that this phage-displayed peptide represents an immunological equivalent of the MGr2 epitope on p185^HER2 and might be used as a substitute for this oncoprotein in in vitro and in vivo immunological studies.     

"Affinity maturation" of ligands for HCV-specific serum antibodies

We have previously screened a phage-displayed random peptide library using sera from patients and identified ligands binding to antibodies specifically associated with the hepatitis C virus infection. The ability of these peptides to detect HCV-specific antibodies was improved through an in vitro procedure which mimics the natural process of antibody affinity maturation operating in secondary immune response. Libraries were generated by mutating the sequence of the original peptide through a protocol that efficiently introduced substitution, insertion and deletion mutations on a single or population of clones. Screening these libraries isolated mutants that displayed increased specific reactivity with a broader range of sera from HCV-infected patients. Several variants of the original peptide were identified which discriminate between the various components of the specific polyclonal response. This methodology to select artificial ligands from RPL using sera and to enhance their diagnostic properties by affinity maturation makes the development of a diagnostic assay to detect disease-associated antibodies feasible, without requiring the natural antigen.     

Detection of hidden rhythms in surface EMG signals with a non-linear time-series tool

The analysis of the surface electromyographic (sEMG) signal is particularly attractive because it provides relatively easy access to those physiological processes that allow the muscle to generate force and movement. In this paper, one of the possible applications of recurrence plot strategy to the analysis of sEMG is described. Recurrence Quantification Analysis (RQA) is an efficient time-series analysis tool pertaining to the class of non-linear dynamics time-domain processing. We analysed sEMG recorded on the biceps brachii during isometric contraction both at constant (CF) and non constant force (NCF). For comparison purposes, experimental data were analysed over epochs of 1 s so that the hypothesis of sEMG stationarity could be accepted. The analysis concerned one of the most widely used frequency parameters (namely the median frequency, MDF) and one parameter (i.e., the percent determinism %DET) extracted using the non-linear technique. Our main results are: (i) the gross average evaluated for all subjects on %DET data shows a comparable variation with respect to MDF throughout the course of CF experiments; (ii) %DET seems able to detect motor unit (MU) synchronisation; (iii) during non constant force experiments, %DET is more effective than MDF in detecting sEMG changes determined by brisk transients of force output.     

nm23 and p53 expression

Breast cancer is a common malignancy and the modalities and the aggressiveness of treatment strategies are variable and depend on information regarding the biological characteristics and behavior of an individual tumor. Therefore, to improve overall survival it is important to identify and select lymph node negative patients at high risk who would benefit from adjuvant therapy. Besides prognostic factors such as lymph node status, hormone receptor status and histopathologic parameters, nm23 antimetastatic gene and the p53 protein were studied in 32 breast cancer patients. Positive staining for nm23 was inversely associated with lymph node involvement in 82.4% of the cases. Most of the non-diploid lesions (70.8%) showed a high protein expression. Positive immunostaining for p53 was present in 28.1% of the cases and was strongly correlated with prognostic indicators such as necrosis and histologic grading. Tumor grade, DNA ploidy and lymph node metastasis were not significantly correlated with p53 protein expression. In this study, all the prognostic indicators studied, satisfactorily explain the important characteristics of the biologic behaviour of breast cancer, but the detection of lymph node metastasis is still the most accurate prognostic factor utilized for a predictive role in disease recurrence.