Novel automated system for magnetic resonance imaging quantification of the inflamed synovial membrane volume in patients with juvenile idiopathic arthritis.

OBJECTIVE: To introduce a novel automated method for the quantification of the inflamed synovial membrane volume (SV) using magnetic resonance imaging (MRI), and to investigate its feasibility and validity in patients with juvenile idiopathic arthritis (JIA). METHODS: The tool was tested on 58 patients with JIA and wrist involvement. Thirty-six patients had a 1-year MRI followup. MRI of the clinically more affected wrist was performed using a 1.5T scanner and a Flex small coil. An algorithmic approach, based on supervised voxel classification for automatic estimation of SV in a 3-dimensional MRI, was developed. The SV was estimated as the number of positively classified voxels and then normalized by the patient's body surface (NSV). Validation procedures included the analysis of reliability, construct validity, responsiveness to change, discriminant validity, and the predictive value. RESULTS: The agreement between the automated estimation of NSV and the manual measurements was excellent (intraclass correlation coefficient 0.93, 95% confidence interval 0.79-0.98). The automatic NSV demonstrated good construct validity by yielding strong correlations with local signs of disease activity and a moderate correlation with global physician assessment of disease activity and with the Rheumatoid Arthritis Magnetic Resonance Imaging Scoring system synovitis score. NSV showed a strong responsiveness to clinical change (standardized response mean values >1) and satisfactory discriminant validity. High baseline NSV (>4.6) had high predictive value (100%) with respect to erosive progression. CONCLUSION: The proposed automated method allowed reliable quantification of NSV, which represents a promising imaging biomarker of disease activity in JIA. The automated system has the potential to improve the longitudinal assessment of JIA and to predict progressive joint destruction.

Spectral algorithms for supervised learning.

We discuss how a large class of regularization methods, collectively known as spectral regularization and originally designed for solving ill-posed inverse problems, gives rise to regularized learning algorithms. All of these algorithms are consistent kernel methods that can be easily implemented. The intuition behind their derivation is that the same principle allowing for the numerical stabilization of a matrix inversion problem is crucial to avoid overfitting. The various methods have a common derivation but different computational and theoretical properties. We describe examples of such algorithms, analyze their classification performance on several data sets and discuss their applicability to real-world problems.

Psychotic disorder and focal epilepsy in a left-handed patient with chromosome XYY abnormality.

OBJECTIVE: To discuss the relationship between XYY chromosome abnormality, psychiatric disorders and epilepsy. METHOD: Single case report. RESULTS: A 34-year-old man with 47, XYY karyotype and normal intelligence was followed-up at a neuropsychiatric clinic for over 30 years. He was first seen at age 3 years with a history of delayed motor and language development and an immunodeficiency syndrome. At age 8 years he developed refractory focal epilepsy, and in late adolescence he started to exhibit increasingly prominent obsessive thoughts, paranoid ideation, and aggressive sexual fantasies and behaviour. CONCLUSIONS: When interpreted within the context of previous literature reports, this case suggests a pathophysiological link between XYY chromosome abnormality, characteristic psychiatric symptoms and epilepsy disorder.

Late diagnosis in severe and mild intellectual disability in adulthood.

BACKGROUND: The diagnosis of intellectual disability (ID) is highly dependent on a comprehensive personal and family medical history, a complete physical examination and a careful developmental assessment of the patient. Our study intended to: (1) classify the aetiology of mild and severe ID in an adult population of 140 Italian subjects; (2) evaluate the frequency of associated medical conditions; (3) evaluate the age of diagnosis in both groups; and (4) underline the importance of aetiological diagnosis for adult ID patients also. METHODS: The study involved 140 consecutive adult Italian ID inpatients and outpatients neurologically investigated at the Neurological Institute C. Mondino of Pavia Service for Mental Retardation. A total of 80 patients had mild ID (MID group) (39 females, 41 males), mean age 34 years (range 19-61 years), mean IQ = 64 (range 51-75), and 60 had severe ID (SID group) (32 females, 28 males), mean age 30 years (range 19-69 years). They underwent a complete diagnostic work-up that comprised prenatal, perinatal and postnatal history, physical examinations, laboratory investigations, genetic survey and neuroradiological investigations to determine the aetiology of ID and to evaluate the presence of associated medical conditions. RESULTS: ID aetiology was classified as prenatal in 34% of the MID and 28% of the SID group. Perinatal and postnatal events were found in 6% of the MID and in 5% of the SID group. Associated medical conditions were found in 97 patients (47% MID and 26% SID). A genetic diagnosis was possible in 6% of patients above 20 years of age and in 5% of patients above 40 years. A diagnosis of cerebral dysgenesis was possible in 5% of patients above 20 years and 4% of patients above 40 years. CONCLUSIONS: A long interval between the diagnosis of ID and the aetiological definition can be observed in a significant percentage (24%) of our population, leading to unfortunate consequences of late diagnosis: late onset of a specific therapeutic program, genetic counselling that is frequently no more useful, and ineffective prenatal diagnosis, leading to the birth of other affected subjects (for familiar ID).

Synthesis and in vitro activity of D- and L-enantiomers of 5-(trifluoromethyl)uracil nucleoside derivatives.

Recently, beta-L-nucleoside analogues have emerged as a new class of sugar modified nucleosides with potential antiviral and/or antitumoral activity. As a part of our ongoing research on this topic, we decided to synthesize 5-CF3-beta-L-dUrd (7), the hitherto unknown L-enantiomer of Trifluridine, an antiherpetic drug approved by FDA but only used in topical applications due to concomitant cytotoxicity. 5-CF3-beta-L-dUrd (7) as well as some other related L-nucleoside derivatives were stereospecifically prepared and tested in vitro against viral (HSV-1 and HSV-2) and human thymidine kinases (TK).

5-(Trifluoromethyl)-beta-l-2'-deoxyuridine, the L-enantiomer of trifluorothymidine: stereospecific synthesis and antiherpetic evaluations.

As a part of our ongoing work on beta-L-nucleoside analogues as potential antiviral drugs, we have synthesized 5-(trifluoromethyl)-beta-L-2'-deoxyuridine (L-TFT), the hitherto unknown L-enantiomer of trifluorothymidine (CF(3)dUrd, TFT). We have also studied the effect of L-TFT on human and herpes simplex virus (HSV) type 1 and 2 thymidine kinases, and human thymidine phosphorylase, as well as its anti-HSV-1 and anti-HSV-2 activities in cell cultures. L-TFT has been found: (i) to inhibit HSV-1 TK with activity comparable to TFT, with no effect on human TK, (ii) to be phosphorylated by the viral enzyme with similar efficiency to TFT, (iii) to be resistant, in contrast to TFT, to hydrolysis by human thymidine phosphorylase. Unfortunately, when evaluated in cell cultures, L-TFT did not show any anti-HSV-1 and anti-HSV-2 activities.

[Suicidal behavior in the North Varese province: a epidemiological analysis]. / La condotta suicidaria nel nord della provincia di Varese: un'indagine epidemiologica.

OBJECTIVE: Analysis of figures and characteristics of suicide behaviour in the area of Varese and the neighbouring northern towns (Valceresio, the valleys around Luino and northern Verbano), with the purpose of finding out preventing measures. DESIGN: Epidemiological-descriptive survey. The 1995-1997 ISTAT death cards, included in the death files of the former USSL of Varese have been taken into consideration. Only those people who were resident at the suicide moment have been included in the survey. Afterwards the psychiatric service archives were consulted, to find any possible contact between the subjects in the ISTAT files and the psychiatric services themselves. With regard to these patients, some information such as suicide attempts, psychiatric pathology and first contact with the psychiatric services have been pointed out. SETTING: Arcisate, Cittiglio, Luino and Varese districts, where the Community Mental Health Services 1 and 2 of "Azienda Ospedaliera Universitaria Macchi" of Varese operate. MAIN OUTCOME MEASURES: The suicide rates in the studied area have been reckoned and, through direct standardization, the rates of the single districts have been compared. RESULTS: There were 78 suicides (24 females and 54 males): rate of 8.2 per 100,000. In accordance with the national trend, there is evidence of a general reduction of the suicide phenomenon, except among the youngest. Districts of Luino and Arcisate are geographic areas to examine in the time, because a wider sample could reveal a higher risk. Young and elderly people are the most affected among the males and, 55/64-year-old people, both males and females, are at higher risk. Suicide methods vary with the age. A high percentage of subjects are not married and with low education. Less than one third of the subjects had come into contact with the psychiatric service. CONCLUSIONS: These data allow a comparison with the national survey and an analysis of the suicide features in the study area. The purpose is to elaborate preventing strategies with multi-dimensional approach whose efficacy may be proved in the future by setting up a provincial observatory.

Riboflavin phosphorylation is the crucial event in riboflavin transport by isolated rat enterocytes.

In isolated rat enterocytes, both normoenergized (normal) and de-energized with rotenone, riboflavin intracellular metabolic processes, operating in association with a membrane-specific transport mechanism, were investigated. The contents of unlabeled (endogenous) and labeled (exogenous) flavins [riboflavin (RF), flavin mononucleotide (FMN), flavin adenindinucleotide (FAD)] were determined by HPLC before and after incubation with tritiated RF. In normoenergized enterocytes, total labeled RF content (i.e., total uptake, the sum of RF membrane transport and intracellular metabolism) increased steadily to a plateau after 20 min incubation; FMN and FAD contents reached a plateau between 3 and 20 min, whereas free RF content increased constantly. The phosphorylated forms prevailed over the free form ( approximately 60% of total flavins). In de-energized enterocytes, RF total uptake was significantly lower than in normoenergized enterocytes and reached a plateau after only 3 min incubation. FMN and FAD contents were significantly lower than in normoenergized enterocytes, and free RF represented the prevailing form of flavins (70% of total RF ). In both normoenergized and de-energized enterocytes, the contents of unlabeled total RF, FMN and FAD decreased significantly after 20 min incubation, whereas free RF increased significantly only in normoenergized enterocytes. After 20 min incubation, the RF structural analog 8-dimethyl-amino-8-demethyl-RF caused a significant decrease of all flavin contents, whereas 5'-deoxy-RF decreased only the total and free RF contents. Results directly confirmed the leading role of metabolic processes such as phosphorylation in RF transport by isolated small intestinal enterocytes.

Anti-(herpes simplex virus) activity of 4'-thio-2'-deoxyuridines: a biochemical investigation for viral and cellular target enzymes.

The antiviral activity of several nucleoside analogues is often limited by their rapid degradation by pyrimidine nucleoside phosphorylases. In an attempt to avoid this degradation, several modified nucleosides have been synthesized. A series of 4'-thio-2'-deoxyuridines exhibits an anti-[herpes simplex virus (HSV)] activity significantly higher (20-600 times) than that shown by the corresponding 4'-oxy counterpart. We investigated the mode of action of these compounds and we found that: (i) several 4'-thio-2'-deoxyuridines are phosphorylated to the mono- and di-phosphates by HSV-1 thymidine kinase (TK) more efficiently than their corresponding 4'-oxy counterpart; (ii) both are inhibitors of cellular thymidylate synthase; (iii) 4'-thio-2'-deoxyuridines are resistant to phosphorolysis by human thymidine phosphorylase; (iv) both 4'-oxy- and 4'-thio-2'-deoxyuridines are phosphorylated to deoxyribonucleotide triphosphate in HSV-1-infected cells and are incorporated into viral DNA; (v) 4'-thio-2'-deoxyuridines are better inhibitors than their 4'-oxy counterparts of [(3)H]thymidine incorporation in HSV-1-infected cells; (vi) 4'-thio-2'-deoxyuridines are not recognized by HSV-1 and human uracil-DNA glycosylases. Our data suggest that 4'-thio-2'-deoxyuridines, resistant to pyrimidine phosphorylase, can be preferentially or selectively phosphorylated by viral TK in HSV-infected cells, where they are further converted into triphosphate by cellular nucleotide kinases. Once incorporated into viral DNA, they are better inhibitors of viral DNA synthesis than their corresponding 4'-oxy counterpart, either because they are not recognized, and thus not removed, by viral uracil-DNA glycosylase, or because they preferentially interfere with viral DNA polymerase.

Transport of thiamin in rat renal brush border membrane vesicles.

BACKGROUND: Unlike intestinal absorption, renal transport of thiamin has received little attention. This study was designed to investigate the reabsorptive steps of thiamin transport in brush border membrane vesicles (BBMVs) from rat kidney proximal tubules using tritiated thiamin with a high specific activity. METHODS: BBMVs prepared from the cortex kidney of rats were suspended in different media, controlling the composition of the intravesicular fluid by prolonged equilibration at 4 degrees C in preincubation buffers of desired composition. Then they were held on ice until used, when they were warmed at 25 degrees C for the uptake experiments. The amount of radioactivity taken up by the vesicles was measured radiometrically after separation with a rapid-filtration procedure. RESULTS: The time course profile of thiamin uptake was Na+ independent; 53% of thiamin taken up was membrane bound. The concentration curve had a biphasic course that was nonlinear (saturable) at physiological concentrations (<1.25 micromol/L) and linear at higher ones. Thiamin uptake was stimulated several-fold by an outwardly directed H+ gradient (pHin 6:pHout 7.5), which caused a transient accumulation of thiamin inside BBMVs against a concentration gradient. The enhanced thiamin uptake was only due to the H+ gradient, which made thiamin binding virtually negligible compared with translocation, and maintained the biphasic course of the concentration curve. The saturable component, however, had kinetic constants 12-fold higher than those in the absence of gradient. Moreover, the saturable component was inhibited by nonlabeled thiamin and its structural analogues, by inhibitors of intestinal thiamin/H+, renal guanidine/H+, and Na+/H+ antiporters, while it remained unmodified by some typical organic cations transported in renal BBMVs. CONCLUSION: The results provide strong evidence for the presence in renal BBMVs of a thiamin/H+ antiport having a 1:1 stoichiometric ratio.

Oral self-mutilation in a patient with rhombencephalosynapsys.

Rhombencephalosynapsis (RS) is a rare cerebellar malformation. Its essential features are the absence of the incisura cerebelli posterior, fusion of the cerebellar hemispheres, the absence of the velum medullare anterius and nuclei fastigii, and fusion of the dentate nuclei, which are shifted towards the mid-line. Clinically, affected patients present with signs of cerebellar and motor disturbances. The present report describes a new patient affected by RS. The subject first presented at the age of 22 years because of a psychiatric symptomatology which was characterized by obsessive oral self-mutilation associated with an intellectual disability. Objective evaluation documented dysmorphic features, while neurological examination showed only a slight truncal ataxia. The subject's IQ was 74 on the Wechsler Scale (verbal IQ = 79, performance IQ = 74). Psychiatric evaluation with DSM-IV criteria documented an obsessive-compulsive personality disorder associated with emotional instability and oral self-mutilation. The typical picture of rhombencephalosynapsis was evident on magnetic resonance imaging. Both chromosomal analysis and routine biochemical investigations were normal. The relationship between oral self-injurious behaviour and cerebellar malformations is discussed with particular regard to the behavioural aspects of cerebellar congenital pathology in affective disorders and in autism.

Synthesis and molecular modeling of novel HSV1 uracil-DNA glycosylase inhibitors.

In a recent paper the first selective inhibitors of HSV1 uracil-DNA glycosylase (UDG) acting in the micromolar range have been reported. A 28.5 kDa catalytic fragment of HSV1 UDG has been crystallized in the presence of uracil, and the structure was recently solved. Starting with the optimized model of binding between 6-(4'-n-octylanilino)uracil (octAU) and UDG some new derivatives have been predicted to be active. In vitro studies with the novel synthetized compounds confirm the plausibility of the model and define the structure features for UDG inhibitors.

Interaction of beta-L-adenosine-5'-triphosphate (L-ATP) with human deoxycytidine kinase, human DNA primase and T4 DNA ligase: does the chance direct enzymatic enantioselectivity?

We demonstrate that L-ATP: 1) as well as its natural D-enantiomer, acts as a phosphate donor in the reaction catalysed by human deoxycytidine kinase; 2) inhibits human DNA-primase and the ATP-dependent T4 DNA ligase. Thus, the lack of enantioselectivity of the enzymes is more frequent than it was believed a few years ago and we suggest that it would depend on chance more than on an evolutionary strategy.

Molecular modeling and synthesis of inhibitors of herpes simplex virus type 1 uracil-DNA glycosylase.

We recently reported the properties of the first selective inhibitors of herpes simplex virus type 1 (HSV1) uracil-DNA glycosylase (UDG), an enzyme of DNA repair that has been proposed to be required for reactivation of the virus from latency. 6-(4-Octylanilino)uracil (octAU) was the most potent inhibitor among a series of 6-(4-alkylanilino)uracils, acting in the micromolar range and without effect against human UDG. A 28.5-kDa catalytic fragment of HSV1 UDG has been crystallized in the presence of uracil, and the structure was recently solved. We have used the coordinates of this structure in order to study interaction of our inhibitors with the enzyme, and a model of binding between octAU and UDG has been derived. Starting with the optimized model, the activity of several octAU analogues was predicted, and the values compared favorably with experimental results found for the synthetic compounds. Several hydrophilic derivatives were predicted and found to be active as UDG inhibitors. These compounds will be useful to determine if UDG, like the viral thymidine kinase, is required for reactivation of HSV1 from latency in nerve cells.

Contingent negative variation indicates phasic arousal for externally cued unilateral eye blink in human subjects.

The contingent negative variation (CNV) as a slow cortical potential was used to investigate cortical processing of externally cued, voluntary unilateral eye blink. Probands blinked as a response within a modified two-stimulus reaction time paradigm. Reaction time and amplitudes of CNV were determined. The activity of the orbicularis oculi muscles (OO) was registered by surface electromyography (EMG). Probands who performed unilateral eye blinks with accurately inhibiting contralateral OO activity showed a significantly higher negativity of the early CNV component compared with the bilateral eye blink condition. This effect was confined to the beginning of unilateral blinking performance. It is suggested that the unilateral eye blink is a challenging motor task, initially requiring an increased cortically driven arousal and attention as revealed by increased early CNV components.

Psychiatric comorbidity and psychosocial stress in patients with tension-type headache from headache centers in Italy. The Italian Collaborative Group for the Study of Psychopathological Factors in Primary Headaches.

A multicenter study was carried out in 10 Italian Headache Centers to investigate the prevalence of psychosocial stress and psychiatric disorders listed by the IHS classification as the "most likely causative factors" of tension-type headache (TTH). Two hundred and seventeen TTH adult outpatients consecutively recruited underwent a structured psychiatric interview (CIDI-c). The assessment of psychosocial stress events was carried out using an ad hoc questionnaire. The psychiatric disorders that we included in the three psychiatric items of the fourth digit of the IHS classification were depressive disorders for the item depression, anxiety disorders for the item anxiety, and somatoform disorders for the item headache as a delusion or an idea. Diagnoses were made according to DSM-III-R criteria. At least one psychosocial stress event or a psychiatric disorder was detected in 84.8% of the patients. Prevalence of psychiatric comorbidity was 52.5% for anxiety, 36.4% for depression, and 21.7% for headache as a delusion or an idea. Psychosocial stress was found in 29.5% of the patients and did not differ between patients with and without psychiatric comorbidity. Generalized anxiety disorder (83.3%) and dysthymia (45.6%) were the most frequent disorders within their respective psychiatric group. The high prevalence of psychiatric disorders observed in this wide sample of patients emphasizes the need for a systematic investigation of psychiatric comorbidity aimed at a more comprehensive and appropriate clinical management of TTH patients.

L-ATP is recognized by some cellular and viral enzymes: does chance drive enzymic enantioselectivity?

We demonstrate that l-ATP is recognized by some enzymes that are involved in the synthesis of nucleotides and nucleic acids. l-ATP, as well as its natural d-enantiomer, acts as a phosphate donor in the reaction catalysed by human deoxycytidine kinase, whereas it is not recognized by either enantioselective human thymidine kinase or non-enantioselective herpes virus thymidine kinase. l-ATP strongly inhibits (Ki 80 microM) the synthesis of RNA primers catalysed by DNA primase associated with human DNA polymerase alpha, whereas RNA synthesis catalysed by Escherichia coli RNA polymerase is completely unaffected. Moreover, l-ATP competitively inhibits ATP-dependent T4 DNA ligase (Ki 25 microM), suggesting that it interacts with the ATP-binding site of the enzyme. Kinetic studies demonstrated that l-ATP cannot be used as a cofactor in the ligase-catalysed joining reaction. On the other hand, l-AMP is used by T4 DNA ligase to catalyse the reverse reaction, even though a high level of intermediate circular nicked DNA molecules accumulates. Our results suggest that a lack of enantioselectivity of enzymes is more common than was believed a few years ago, and, given the absence of selective constraints against l-nucleosides in Nature, this may depend on chance more than on evolutionary strategy.

An Italian-Australian comparison of quality of life among people with intellectual disability living in the community.

This paper reports the measurement of both objective and subjective quality of life in samples of people either with intellectual disability or from the general adult population, drawn from Australia and Italy. Measures were made using the Comprehensive Quality of Life Scale. Generally, it was found that the scores for all groups were comparable. This finding is consistent with the provision of generally decent objective living conditions in both countries and subjective well-being homeostasis, wherein subjective quality of life is held within a predictable range. These results are discussed in the context of measurement difficulties imposed by the need for simple Likert scales and subjective data which are strongly negatively skewed.

Cerebellar dysgenesis and mental retardation associated with a complex chromosome rearrangement.

Cerebellar hypoplasia, mild mental retardation, skeletal abnormalities, and ataxia were present in a 40 years old patient with a complex chromosome rearrangement (CCR). Chromosomes 2, 5, 16, and 17 were involved in the CCR. For the definition of the eight breakpoints leading to the rearrangement FISH with whole chromosomes paintings and specific telomeric probes was employed. Gene disruption, positional effect variegation, and sub-microscopic deletions are all possible causes for the abnormal phenotype observed in the patient.

Neurological symptoms of the sick building syndrome: analysis of a questionnaire.

Sick building syndrome (SBS) is an excess of work-related irritations of the skin and mucous membranes and of symptoms such as headache and fatigue in those working in modern air-conditioned buildings. We aimed to analyse the neurological symptoms, especially headache, in workers with potential SBS. The most frequent symptoms were headache and dry eyes. Sex was a major factor of difference: women report more symptoms than men. A positive correlation emerged between the number of symptoms and the asthenia scale score. Only 11 (8.2% of the whole sample) and 37 (27.4%) workers met all the IHS criteria for migraine and tension-type headache respectively. At least one symptom of SBS was present in 92.6% of workers. A negative correlation emerges between air conditioning and headache during working hours. No correlation emerges between the workplace comfort indicator and SBS and asthenic symptoms while a negative correlation was found between migraine and tension-type headache and comfort in the workplace. SBS symptoms are very frequent among all workers but headache is the primary symptom.