Localization of neuroglobin in the brain of R6/2 mouse model of Huntington's disease.

Neuroglobin (Ngb) is expressed in the central and peripheral nervous system, cerebrospinal fluid, retina, and endocrine tissues where it is involved in binding O2 and other gasotransmitters. Several studies have highlighted its endogenous neuroprotective function. Huntington's disease (HD), a dominant hereditary disease, is characterized by the gradual loss of neurons in discrete areas of the central nervous system. We analyzed the expression of Ngb in the brain tissue of a mouse model of HD, in order to define the role of Ngb with respect to individual cell type vulnerability in HD and to gender and age of mice. Our results showed different expressions of Ngb among neurons of a specific region and between different brain regions. We evidenced a decreased intensity of Ngb at 13 weeks of age, compared to 7 weeks of age. The double immunofluorescence and fluorescence resonance energy transfer (FRET) experiments showed that the co-localization between Ngb and huntingtin at the subcellular level was not close enough to account for a direct interaction. We also observed a different expression of Ngb in the striatum, depending on the sex and age of animals. These findings provide the first experimental evidence for an adaptive response of Ngb in HD, suggesting that Ngb may exert neuroprotective effects in HD beyond its role in reducing sensitivity to oxidative stress.

Miglustat Reverts the Impairment of Synaptic Plasticity in a Mouse Model of NPC Disease.

Niemann-Pick type C disease is an autosomal recessive storage disorder, characterized by abnormal sequestration of unesterified cholesterol within the late endolysosomal compartment of cells and accumulation of gangliosides and other sphingolipids. Progressive neurological deterioration and insurgence of symptoms like ataxia, seizure, and cognitive decline until severe dementia are pathognomonic features of the disease. Here, we studied synaptic plasticity phenomena and evaluated ERKs activation in the hippocampus of BALB/c NPC1-/- mice, a well described animal model of the disease. Our results demonstrated an impairment of both induction and maintenance of long term synaptic potentiation in NPC1-/- mouse slices, associated with the lack of ERKs phosphorylation. We then investigated the effects of Miglustat, a recent approved drug for the treatment of NPCD. We found that in vivo Miglustat administration in NPC1-/- mice was able to rescue synaptic plasticity deficits, to restore ERKs activation and to counteract hyperexcitability. Overall, these data indicate that Miglustat may be effective for treating the neurological deficits associated with NPCD, such as seizures and dementia.

miR-34a regulates cell proliferation, morphology and function of newborn neurons resulting in improved behavioural outcomes.

miR-34a is involved in the regulation of the fate of different cell types. However, the mechanism by which it controls the differentiation programme of neural cells remains largely unknown. Here, we investigated the role of miR-34a in neurogenesis and maturation of developing neurons and identified Doublecortin as a new miR-34a target. We found that the overexpression of miR-34a in vitro significantly increases precursor proliferation and influences morphology and function of developing neurons. Indeed, miR-34a overexpressing neurons showed a decreased expression of several synaptic proteins and receptor subunits, a decrement of NMDA-evoked current density and, interestingly, a more efficient response to synaptic stimulus. In vivo, miR-34a overexpression showed stage-specific effects. In neural progenitors, miR-34a overexpression promoted cell proliferation, in migratory neuroblasts reduced the migration and in differentiating newborn neurons modulated process outgrowth and complexity. Importantly, we found that rats overexpressing miR-34a in the brain have better learning abilities and reduced emotionality.

Prognostic outcome of routine clinical noninvasive multidetector-row computed tomography coronary angiography in patients with suspected coronary artery disease: a 2-year follow-up study.

PURPOSE: The aim of the study was to assess the prognostic value of multidetector-row CT coronary angiography (MDCT-CA) in patients with suspected coronary artery disease (CAD) in a routine clinical context. MATERIALS AND METHODS: A total of 125 patients (82 men, age 57.4±10.3 years) with suspected CAD underwent MDCT-CA. All patients were assessed for cardiovascular risk factors, symptoms and coronary calcium score. A 2-year follow-up study for the occurrence of major adverse cardiac events was performed. RESULTS: According to the Morise pretest score, 76 patients (60.8%) were at intermediate risk. Patients with suspected CAD presented the following prognostic outcome (p<0.0001): in 41 patients with normal coronary arteries at MDCT-CA, the event rate was 0%; five of 49 patients with nonobstructive CAD had major cardiac events; two of 35 patients with obstructive CAD suffered cardiac death and 19 underwent revascularisation. At multivariate analysis, the presence of obstructive CAD is the only significant independent prognostic variable (hazard ratio, 10.1393; 95% confidence interval 3.2189-31.9379; p<0.0001). CONCLUSIONS: Routine clinical MDCT-CA provides an excellent prognostic value at 2-year follow-up in patients with normal coronary arteries. The cardiac event rate increases with CAD severity.

Prevalence of myocardial bridging and correlation with coronary atherosclerosis studied with 64-slice CT coronary angiography.

PURPOSE: This study aimed to assess the prevalence and characteristics of myocardial bridging in patients who underwent multislice computed tomography coronary angiography (MSCT-CA) and to evaluate the correlation between bridged coronary segments and atherosclerosis. MATERIALS AND METHODS: A total of 277 patients (mean age 60+/-11 years) we consecutively examined with 64-slice MSCT-CA for suspected or known coronary atherosclerosis were retrospectively reviewed for myocardial bridging. Segments proximal and distal to the bridging were evaluated for atherosclerotic plaque, as were the remaining coronary segments. RESULTS: Myocardial bridging was present in 82 patients (30%, mean age 59+/-12). Bridges were of variable length (<1 cm 58%; 1-2 cm 32%; >2 cm 10%) and depth (superficial 69%, intramyocardial 31%) and frequently localised in the mid-distal segment of the left anterior descending artery (95%). Myocardial bridging cannot be considered a significant risk factor for coronary atherosclerosis (odds ratio 0.49) compared with traditional cardiovascular risk factors. Coronary segments proximal to the bridge showed no atherosclerotic disease (33%), positive remodelling (27%), <50% stenosis (20%) or >50% stenosis (20%). We identified 12 noncalcified, 32 mixed and 17 calcified plaques. The distal segments were significantly less affected (p<0.0001). CONCLUSIONS: MSCT-CA is a reliable, noninvasive method that is able to depict myocardial bridging and associated atherosclerotic plaque in the proximal segments.

[Evaluation of operating theatre for the risk prevention]. / Valutazione della qualità della Sala Operatoria in funzione della prevenzione del rischio.

In this study we analyse factors that predispose to risk and we value the importance of quality and reliability into operating theatre. Patient safety result from ability to plan and manage organizations able to reduce probability of mistakes (Prevention) and to recover and contain their consequences (Protection). The principal motives of mistakes are: - deficiency of sharing procedures between different professional figures for risk prevention; - deficiency of an effective integration between professional figures in operating theatre; - deficiency of charitable precise run to guarantee the continuity of interventions on patient. A risk management program have to take care on this sentinel events and to set up a survey-information system to characterize risks and correction strategies. To prevent patient change, wrong side identification, not working devices or deficiency of surgery materials, it would be useful for operating theatre figures and for anesthetists to work out all together a procedure for admission in operating theatre and a pre-operating check list. The best way to carry out a safety and quality attendance based on standardized procedures and protocols, is to set up risk management firm units. It is necessary to activate and to adjust survey system and effective management training.

[The hospital: reality and proposable future]. / L'Ospedale: realtà e futuro proponibile.

In this work we run over hospital history from Middle ages to the present time. Hospitals were charitable institutions in fact they rise inside monasterys, churches or castles with an architecture in modum crucis as a symbolic expression of Christ crucifix. During Renaissance, new scientific Knowledges and the need of technical assistance in a suitable place, lead to a new kind of hospital included into the functional centres of the city were medical practice takes the places of spiritual assistance. In XVIII century hospital is like a "human diseases botanical garden" divided into departments with a circular structure in conformity with a functional model of control. To exceed the isolation of single room, rectangular rooms born with a radially arrangement. At the end of 700's born the pavilions structures typical of hospitals until half 900's when the "monobloc" take place. Today hospital becomes horizontal, include in the context of the city, with hall as a big hotel and with trading centres in accordance with Renzo Piano model and with a new vision of hospital as a welcome place were the patient is a guest to treat as a person of consequence.

Digital cineradiographic study of swallowing in patients with amyotrophic lateral sclerosis.

PURPOSE: This study was performed to evaluate the usefulness of digital cineradiography in detecting swallowing disorders in dysphagic patients affected by amyotrophic lateral sclerosis (ALS) with a view to planning an adequate therapeutic approach. MATERIAL AND METHODS: From January 2005 to September 2006, 23 patients (10 men and 13 women; mean age 41.3+/-8.6 years) affected by ALS were evaluated with digital cineradiography to assess the grade of dysphagia. All patients were classified using the Hillel ALS Severity Scale (ALSSS). All examinations were performed with radiocontrolled equipment provided with a digital C-arm. RESULTS: The cineradiographic technique enabled us to differentiate patients with disorders of the oral (17/23) and/or pharyngeal (19/23) swallowing phase from those without swallowing dysfunction (4/23). In 14/23 patients, passage of contrast medium into the upper airways was observed during swallowing, whereas in 5/23 cases, aspiration of contrast medium into the lower airways was recorded. CONCLUSIONS: The videofluoroscopic swallowing study has high diagnostic capabilities in the evaluation of swallowing disorders, as it is able to identify the degree and causes of impairment. In addition, the study proved useful for planning speech therapy and for follow-up in patients with ALS.

Heterogeneous delayed enhancement of hepatic parenchyma after intravenous infusion of sonographic contrast agent: a new hypothesis.

PURPOSE: The aim of this paper was to report a heterogeneous late-phase hepatic enhancement pattern observed after administration of a sonographic contrast agent and to present an aetiological hypothesis for the phenomenon. MATERIALS AND METHODS: A total of 1,729 (1,012 women and 717 men; age range 28-82; mean age 51) patients underwent contrast-enhanced sonography of the liver. The examination was performed with a low mechanical index (MI <0.09) after injection of sulphur-hexafluoride-filled microbubbles, using different sonographic equipment and different contrast-specific algorithms. RESULTS: Heterogeneous delayed liver enhancement was observed in six patients in the late phase (180 s), with the presence of multiple, partially confluent, hyperechoic areas peripheral to the portal vessels. The pattern appeared spontaneously between 1 and 4 h after the examination and was associated with the presence of an anomalous echogenicity in the superior mesenteric vein. No patient experienced adverse reactions. CONCLUSIONS: The phenomenon of heterogeneous hepatic enhancement may be related to gas from the intestinal microcirculation being transported to via the enteroportal circulation and becoming trapped in the hepatic sinusoids.

Early-onset progressive ataxia associated with the first CACNA1A mutation identified within the I-II loop.

Familial hemiplegic migraine type 1, spinocerebellar ataxia type 6 (SCA6) and episodic ataxia type 2 (EA2) are allelic disorders associated with mutations in the CACNA1A gene, which encodes the alpha1 subunit of the P/Q-type calcium channel (Ca(V)2.1). SCA6 and EA2 share a number of clinical features, such as prominent cerebellar involvement and good response to acetazolamide therapy. However, while SCA6 develops as a late-onset, progressive ataxia, EA2 has an earlier, and episodic, onset. We report on two sisters with a heterogeneous clinical phenotype. The first developed progressive cerebellar ataxia after age 30, without noticeable episodes of vertigo or headache. A 1 year trial with acetazolamide did not produce significant results. The other reported episodes of vertigo, headache and gait imbalance since late childhood, with good response to acetazolamide, before developing moderate chronic cerebellar ataxia. Brain MRI showed cerebellar atrophy, especially in the vermis, in both patients. Direct sequencing of CACNA1A identified a heterozygous 1360G>A mutation in exon 11 resulting in the substitution of alanine for threonine at residue 454 (p.Ala454Thr). This is the first description of a change residing in the cytoplasmic I-II loop associated with a clinical phenotype.

Reduced GABAB receptor subunit expression and paired-pulse depression in a genetic model of absence seizures.

Neocortical networks play a major role in the genesis of generalized spike-and-wave (SW) discharges associated with absence seizures in humans and in animal models, including genetically predisposed WAG/Rij rats. Here, we tested the hypothesis that alterations in GABA(B) receptors contribute to neocortical hyperexcitability in these animals. By using Real-Time PCR we found that mRNA levels for most GABA(B(1)) subunits are diminished in epileptic WAG/Rij neocortex as compared with age-matched non-epileptic controls (NEC), whereas GABA(B(2)) mRNA is unchanged. Next, we investigated the cellular distribution of GABA(B(1)) and GABA(B(2)) subunits by confocal microscopy and discovered that GABA(B(1)) subunits fail to localize in the distal dendrites of WAG/Rij neocortical pyramidal cells. Intracellular recordings from neocortical cells in an in vitro slice preparation demonstrated reduced paired-pulse depression of pharmacologically isolated excitatory and inhibitory responses in epileptic WAG/Rij rats as compared with NECs; moreover, paired-pulse depression in NEC slices was diminished by a GABA(B) receptor antagonist to a greater extent than in WAG/Rij rats further suggesting GABA(B) receptor dysfunction. In conclusion, our data identify changes in GABA(B) receptor subunit expression and distribution along with decreased paired-pulse depression in epileptic WAG/Rij rat neocortex. We propose that these alterations may contribute to neocortical hyperexcitability and thus to SW generation in absence epilepsy.

BIS - AAI and clinical measures during propofol target controlled infusion with Schnider's pharmacokinetic model.

AIM: The A-line autoregressive index (AAI) and the Bispectral Index Score (BIS) are two commercially available indexes of anesthetic depth widely used in clinical practice. The aim of the current study was to compare the accuracy of AAI, BIS, Schnider's predicted effect-site concentration of propofol (Ce propofol) to assess depth of anesthesia. METHODS: Forty-four patients scheduled for major elective abdominal surgery received target effect-site controlled infusion of propofol. Target effect-site (Ce propofol) was started at 1.5 mug/mL and increased every 4 min by 1.0 microg/mL until 5.5 microg/mL were achieved. At every step sedation level was estimated, using AAI, BIS, Observer's Assessment of Alertness/Sedation scale (OAA/S), loss of eyelash reflex and Ce propofol. RESULTS: We enrolled 44 patients, 20 males and 24 females, ASA I/II 18/26, 48+/-10 years, 68.2+/-9 kg, 165+/-7.1 cm, body mass index (BMI) 25+/-3.5. At increasing Ce propofol BIS-AAI values decreased progressively (BIS range 97-38) (AAI range 97-17). Values of BIS < or = 50, of AAI < or = 48 and of Ce propofol > or = 5.1 resulted in OAA/S=0, while values of BIS < or = 62, AAI < or = 53 and Ce propofol < or = 3.5 resulted in OAA/S=2. Loss of eyelash reflex occurred when values were BIS < or = 64 and AAI < or 61. CONCLUSION: BIS, AAI, propofol site effect concentration revealed information on sedation level and consciousness but no gold standard yet exists because of consistent overlap between ''conscious'' and ''not conscious'' states.

Characterization of benign hepatic tumors arising in fatty liver with SonoVue and pulse inversion US.

BACKGROUND: We describe the spectrum of contrast-enhancement patterns of benign hepatic tumors arising in fatty liver on contrast-enhanced ultrasound (US). METHODS: Sixteen patients (12 women and four men) with 27 benign hepatic tumors (17 hemangiomas, eight focal nodular hyperplasias, and two hepatocellular adenomas) arising in fatty liver underwent baseline and pulse inversion US after administration of SonoVue. Two experienced radiologists evaluated baseline echogenicity and dynamic enhancement pattern of each lesion in comparison with adjacent liver parenchyma. RESULTS: After administration of SonoVue, in the arterial phase 13 of 17 hemangiomas showed peripheral globular enhancement and one showed a rim of peripheral enhancement, followed by progressive centripetal fill-in, which was complete in 10 of 14 cases and incomplete in four. Three of 18 hemangiomas showed rapid and complete fill-in in the arterial phase. Eight of eight focal nodular hyperplasias became hyperechoic in comparison with adjacent liver parenchyma in the arterial phase and slightly hyperechoic or isoechoic in the portal venous and delayed phases. Both adenomas showed strong arterial contrast enhancement that became less intense in the portal venous and delayed phases. CONCLUSION: Contrast-enhanced US after administration of SonoVue enables depiction of typical contrast-enhancement patterns in most benign hepatic tumors arising in fatty liver, thus providing useful clues for characterization.

Pancreatic carcinoma versus chronic focal pancreatitis: contrast-enhanced power Doppler ultrasonography findings.

BACKGROUND: We assessed the potential usefulness of contrast-enhanced power Doppler ultrasonography (US) for differentiating pancreatic carcinoma from chronic focal pancreatitis. METHODS: Twenty-six consecutive patients with ductal carcinoma (n=16) and chronic focal pancreatitis (n=10) underwent power Doppler US examinations before and after intravenous injection of an air-based contrast agent (Levovist, Schering, Berlin, Germany). Final diagnosis was obtained by surgery in all patients. The following parameters before and after intravenous administration of contrast agent were evaluated for each lesion: number, morphology and course of the vessels within the lesion and time to maximum detectable enhancement. Number of the lesion vessels and time to maximum enhancement for each lesion were compared by with Student t test. RESULTS: After injection of contrast agent, nine of 16 (56.2%) carcinomas showed a larger total number of vessels (p<0.005) and faster enhancement (p<0.0001) compared with chronic focal pancreatitis. Irregular vessels with a serpiginous course that penetrated the tumor was predominant in carcinoma (eight of 16 cases), whereas focal pancreatitis presented mostly as avascular masses (five of 10 cases). CONCLUSION: Contrast-enhanced power Doppler US provides useful findings for differentiating pancreatic carcinoma from chronic focal pancreatitis.

Pigmented villonodular synovitis of the foot: MR findings.

PURPOSE: To evaluate the MRI findings in the various forms of pigmented villonodular synovitis (PVNS) of the foot. MATERIALS AND METHODS: Seven hundred and fifty-three MR studies of the foot performed at our institute between June 1994 and April 2000 were retrospectively reviewed for the presence of PVNS. Spin echo (SE) T1W, Gradient echo (GE) T2*W, and fat suppression (Short Time Inversion Recovery: STIR) images were acquired with a 0.5 T superconductive unit (Vectra, GE Medical System, Milwaukee, WI, USA) provided with a dedicated transmitter/receiver coil. The site and type of lesions, the signal intensity patterns, and the presence of associated changes were evaluated. RESULTS: On the basis of the MR images and the above parameters, six patients (3 men, 3 women, age range: 35-48 years) with PVNS were selected. Three out of six PVNS were nodular, whereas the remaining three were diffuse. Of the three nodular forms, one was found in the sub-talar joint and the remaining two antero-medially to the talus. Instead, all of the diffuse lesions were located on the metatarsus. Perilesional oedema was seen in all cases, although more obvious in the nodular forms, whereas bone involvement (osteochondral erosion) was observed only in the diffuse metatarsal PVNS. Intra-articular bloody effusion was never observed. The MRI findings were confirmed by surgery in all cases. DISCUSSION: The high contrast resolution and multiplanar capabilities of MRI allow the complete evaluation of the structures of the foot affected by PVNS, and of the extent of soft tissue (bursae, synovial or nervous structures), bone and articular involvement. Although not specific, the presence of haemosiderin results in characteristic MR findings, due to the shortening of both T1 and T2 relaxation times. GET2* images are particularly well suited to this PURPOSE: Furthermore, in our experience, FIR images added better depiction of associated swelling. CONCLUSIONS: According to our results, MRI is now the most reliable technique for identifying and classifying PVNS, and allows correct treatment planning and effective monitoring.

Analysis of radioactive pollution in the Gulf of Palermo.

PURPOSE: Sea water represents more than three quarters of the whole world. The radioactive fall out caused by nuclear experiments, radioactive drosses from nuclear power plants and other pollution sources, are able to contaminate the world ecosystems, and especially the sea water. The aim of this work is the evaluation of a possible radioactive pollution in the Gulf of Palermo. MATERIAL AND METHODS: The Gulf of Palermo was divided into 5 homogeneous hydrological parts, in which fish and water were collected. The dosage of 131I e del 137Cs was mainly assessed because they have a long half-life and their presence in the water can be due to nuclear accidents but also to a wrong removal of the waste materials of nuclear medicine procedures; the other radioisotopes assessed (144Ce, 106Rh, 140La) are due only to nuclear fall-out. Gamma spectrometry technique was used for the evaluation of the radioisotopes' concentration after an adequate treatment of the samples. RESULTS AND CONCLUSIONS: The concentration of the examined radionuclides was below the instrumental capability apart from the 137Cs, whose dose was increased, and can be due to nuclear accidents and explosions. The radionuclide levels detected do not determine a higher risk than normal for the population's health even regard the eatablity of ittical species.

[Sportsman hindfoot pain: role of magnetic resonance imaging]. / Tallodinia dello sportivo: ruolo della risonanza magnetica.

PURPOSE: To assess the outcome of Magnetic Resonance Imaging (MRI) in the diagnosis of sportsman hindfoot pain. MATERIALS AND METHODS: Fortythree professional athletes (31 men, 12 women, age range: 17-37 years) affected by hindfoot pain underwent MRI. Spin echo (SE) T1W, Gradient echo (GE) T2*W, and fat suppression (Short Time Inversion Recovery: STIR) images were acquired with a 0.5 T superconductive unit (Vectra, GE Medical System, Milwaukee, WI, USA). A dedicated extremities transmitter/receiver coil was used. The lesion site, the presence of anatomic variants (os trigonum, Haglund's deformity), and signal intensity changes were evaluated. RESULTS: In all cases MRI allowed the identification of the cause of the hindfoot pain, in relation to soft tissue (bursae, synovial or nervous structures), bone and articular diseases. Particularly, as regards soft tissue diseases, tendinous abnormalities and inflammatory bursal involvement were frequently found (77% of cases). Bone diseases (22% of cases with posterior talalgia alone), mostly involved the heel (60%), whereas cartilagineous diseases were present in 9% of cases. In 60% of cases an intra-articular osseous or cartilagineous displaced fragment coexisted, determinating joint locking during foot flexo-extension movements. In 38% of cases contemporary involvement of different articular structures was observed. DISCUSSION: Both MRI high contrast resolution and multiplanar capabilities allow the complete evaluation of hindfoot region. In our experience sagittal and axial planes were particularly well suited for the diagnosis and the assessment of disease extension. Furthermore, T2W (GET2*) and fat suppression (STIR) images allow high sensitivity even in early disease detection, when hyperemia or fluid collection occur. CONCLUSIONS: According to our results, it seems possible to state that nowadays MRI is the most reliable technique for identifying the causes of hindfoot pain, in order to provide a correct and effective pre-therapeutic planning.

Aggresome formation by anti-Ras intracellular scFv fragments. The fate of the antigen-antibody complex.

Diverting the antigen from its normal intracellular location to other compartments in an antibody-mediated way represents a mode of action for intracellular antibodies [Cardinale, A., Lener, M., Messina, S., Cattaneo, A. & Biocca, S. (1998) FEBS Lett., 439, 197-202; Lener, M., Horn, I.R., Cardinale, A., Messina, S., Nielsen, U.B., Rybak, S.M., Hoogenboom, H.R., Cattaneo, A. & Biocca, S. (2000) Eur J Biochem. 267, 1196-205]. In the case of p21Ras, the sequestration of the antigen in aggregated structures in the cytoplasm of transfected cells leads to the inhibition of its biological function. We have further investigated the intracellular fate of the antigen-antibody complex by analyzing the effect of proteasome inhibitors on the formation and the intracellular localization of the aggregates. Overexpression of anti-Ras scFv fragments or inhibition of proteasomes activity leads to the formation of large perinuclear aggresomes formed of ubiquitinated-scFv fragments in which p21Ras is sequestered and degraded in an antibody-mediated way. Disruption of microtubules by nocodazole completely abrogates the accumulation of scFv fragments in a single aggresome and induces the dispersion of these structures in the periphery of the cell. Cotransfection of the GFP-scFv with a myc-tagged ubiquitin and colocalization with specific anti-proteasome antibodies indicate the recruitment of exogenous ubiquitin and proteasomes to the newly formed aggresomes. Taken together these results suggest that the intracellular antigen-antibody complex is naturally addressed to the ubiquitin-proteasome pathway and that the mechanism of ubiquitination does not inhibit the antibody binding properties and the capacity to block the antigen function.