Hybrid solid-state SPECT/CT left atrial innervation imaging for identification of left atrial ganglionated plexi: Technique and validation in patients with atrial fibrillation.

BACKGROUND: Ablating left atrial (LA) ganglionated plexi (GP), identified invasively by high-frequency stimulation (HFS) during pulmonary vein isolation (PVI), may reduce atrial fibrillation (AF) recurrence. 123I-metaiodobenzylguanidine (123I-mIBG) solid-state SPECT LA innervation imaging (LAII) has the spatial resolution to detect LAGP non-invasively but this has never been demonstrated in clinical practice. METHODS: 20 prospective patients with paroxysmal AF scheduled for PVI underwent 123I-mIBG LAII. High-resolution tomograms, reconstructed where possible using cardiorespiratory gating, were co-registered with pre-PVI cardiac CT. Location and reader confidence (1 [low] to 3 [high]) in discrete 123I-mIBG LA uptake areas (DUAs) were recorded and correlated with HFS. RESULTS: A total of 73 DUAs were identified, of which 59 (81%) were HFS positive (HFS +). HFS + likelihood increased with reader confidence (92% [score 3]). 64% of HFS-negative DUAs occurred over the lateral and inferior LA. Cardiorespiratory gating reduced the number of DUAs per patient (4 vs 7, P = .001) but improved: HFS + predictive value (76% vs 49%); reader confidence (2 vs 1, P = .02); and inter-observer, intra-observer, and inter-study agreement (κ = 0.84 vs 0.68; 0.82 vs 0.74; 0.64 vs 0.53 respectively). CONCLUSIONS: 123I-mIBG SPECT/CT LAII accurately and reproducibly identifies GPs verified by HFS, particularly when reconstructed with cardiorespiratory gating.

Prevalence of Polysubstance Abuse and Dual Diagnosis in Patients Admitted to Alcohol Rehabilitation Units for Alcohol-Related Problems in Italy: Changes in 15 Years.

AIMS: Alcohol abuse has long been known as a disease with social and economic burden to society. Given the complex nature of alcohol treatment, it is worthwhile to examine the change over time of patients admitted to residential alcohol abuse rehabilitation units. METHODS: The data were collected from two Italian projects on alcoholics performed in the mid-1990s (ASSALT) and in 2009 (CORRAL), respectively. Categorical variables were considered in terms of absolute and relative frequencies. Comparisons of relative frequencies between groups were assessed by means of Fisher's exact test. Mixed logistic regression models were fitted to CORRAL data to identify the predictors of the probability of being a polysubstance abuser or having a dual diagnosis. The association estimates were reported as adjusted odds ratios and relative 95% confidence intervals. RESULTS: Compared to the mid-1990s, in 2009 patients were older (P= 0.0003), with a higher level of education (P= 0.0204), with fewer comorbidities (liver disease except cirrhosis, P < 0.0001; polyneuropathy, P= 0.0001), more frequently polysubstance abusers (P < 0.0001), affected by dual diagnosis (P < 0.0001). In 2009, the probability of being a polysubstance abuser was higher in younger and in patients with dual psychiatric diagnosis. Female gender and polysubstance abuse were positively associated to the probability of being affected by dual psychiatric diagnosis. CONCLUSIONS: The increment of patients admitted to residential programs for alcohol dependence with polysubstance abuse and/or dual psychiatric diagnoses suggests the need to pay more attention to both psychological/psychiatric interventions and internal medicine/physical rehabilitation. SHORT SUMMARY: The results of this study suggest that further research is needed to identify the best treatment strategy that is safe and effective for the new population of alcoholics.

Production and characterization of natural rubber-Ca/P blends for biomedical purposes.

This study presents the development of natural rubber-Ca/P blends, as promising candidates for biomedical purposes. The specific objective was the incorporation of Ca/P into a natural rubber polymeric matrix. Ca/P crystalline phases were synthesized by the sol-gel method and the polymeric matrices were produced using natural rubber extracted from latex of the Hevea brasiliensis. The shape and size of natural rubber particles present in the NR membrane, as well as, the way the Ca/P powder grains aggregate in the polymeric matrix were investigated, giving information about the interactions between the Ca/P and the natural rubber particles. Confocal fluorescence scanning microscopy measurements allowed us to propose a structure where the Ca/P grains are surrounded by natural rubber particles. This structure may mediate Ca(2+) release for tissue regeneration. The system investigated may open new horizons for development of a bandage which provides the controlled-release of biomaterials.

[Alcohol-addiction inpatient: characteristics of patients and rehabilitation program]. / La residenzialità alcologica ospedaliera: caratteristiche dei pazienti e dei percorsi terapeutici/riabilitativi.

AIM: The treatment of alcohol addiction in Italy has had a progressive evolution of therapeutic structures for in-and outpatients. During the last 20 years there had been a crescent presence of short residential treatment facilities (1-6 months) characterized by a high level of medical and psychotherapeutic intervention. About two years ago 12 of them jointed together in an association called CORRAL (COordinamento of Residenzialità Riabilitive Alcologiche). The aim of this study was to describe the socio-demographic medical and other characteristics of the patients coming for this type of treatment and to describe the characteristics of the residential treatment itself including referring and aftercare. METHODS: Two thousand sixty-one hospitalized patients of the 12 rehabilitative alcohol units were examined by using a questionnaire and collecting various sociodemographic variables and clinical diagnosis of the patients. Even it was asked who had referred the patients, the characteristics of the residential treatment and of the planned aftercare. RESULTS: The present residential facilities are mainly distributed in the North of Italy. The typical patient is male, with a high school instruction and with a comorbidity regarding psychiatric and liver disorders. The majority of the patients were referred by the public ambulatory services for addictions (SerT/SerD). The characteristics of the residential treatment were medical, pharmacological and psychotherapeutic interventions with a high intensity and the presence of a general-purpose staff. CONCLUSIONS: This study outlined a model of residential rehabilitation of alcohol disorders characterized by short duration and a complex, intense therapeutic intervention mainly addressed to patients with a severe clinical condition and a low level of social problems. Further research should be useful to understand better which sort of patient characteristics obtain a better clinical result and for that even economically a major effectiveness from this type of residential treatment.

Cardiac 123I-MIBG scintigraphy in heart failure.

This review focuses on myocardial [(123)I]mIBG imaging in patients with cardiomyopathy and heart failure (HF). In particular, the potential role of myocardial (123)I-mIBG scintigraphy in the prediction of potentially fatal arrhythmic events is described. A growing body of evidence supports the use of cardiac sympathetic innervation imaging, specifically using (123)I-mIBG, to risk stratify patients with HF. Cardiac sympathetic imaging can help to improve understanding of the mechanisms responsible for increased sympathetic activity in HF, and how sympathetic over-activity exerts its deleterious actions. Eventually, this may result in more effective therapies and, thereby, better outcome for these patients. Assessment of cardiac sympathetic activity could also contribute to a more appropriate selection of individual patients for implantable cardioverter-defibrillators (ICD).

Factors affecting in-hospital heat-related mortality: a multi-city case-crossover analysis.

BACKGROUND: Several studies have identified strong effects of high temperatures on mortality at population level; however, individual vulnerability factors associated with heat-related in-hospital mortality are largely unknown. The objective of the study was to evaluate heat-related in-hospital mortality using a multi-city case-crossover analysis. METHODS: We studied residents of four Italian cities, aged 65+ years, who died during 1997-2004. For 94,944 individuals who died in hospital and were hospitalised two or more days before death, demographics, chronic conditions, primary diagnoses of last event and hospital wards were considered. A city-specific case-crossover analysis was performed to evaluate the association between apparent temperature and mortality. Pooled odds ratios (OR) of dying on a day with a temperature of 30 degrees C compared to a day with a temperature of 20 degrees C were estimated with a random-effects meta-analysis. RESULTS: We estimated an overall OR of 1.32 (95% confidence interval: 1.25, 1.39). Age, marital status and hospital ward were important risk indicators. Patients in general medicine were at higher risk than those in high and intensive care units. A history of psychiatric disorders and cerebrovascular diseases gave a higher vulnerability. Mortality was greater among patients hospitalised for heart failure, stroke and chronic pulmonary diseases. CONCLUSIONS: In-hospital mortality is strongly associated with high temperatures. A comfortable temperature in hospitals and increased attention to vulnerable patients during heatwaves, especially in general medicine, are necessary preventive measures.

Bdnf expression in rat skeletal muscle after acute or repeated exercise.

Brain derived growth factor (BDNF) gene of rat has a complex structure: at least four 5' untranslated exons regulated by different promoters and one 3' exon containing the encoding region. BDNF is expressed by skeletal muscles in an activity-dependent manner. In this study, BDNF mRNA was analysed by RT-PCR in the soleus muscle following a single (acute) session of running or a training of five days of running (repetitive exercise). Moreover, the expression of the exons was quantitatively analysed by real time RT-PCR. Finally, muscle BDNF protein level was evaluated by western blotting. BDNF mRNA was found to increase over the second day after acute exercise; on the other hand, two peaks (2 and 24 hours after the last session, respectively) in BDNF mRNA level were found after repetitive exercise, but it was similar to that of controls 6 hours after the last session. BDNF protein level progressively increased also after the mRNA went back to the basal level, so suggesting that it cumulates within the cell after acute exercise, whereas it followed the mRNA level time course after repetitive exercise. These results point to the following conclusions: BDNF mRNA is up-regulated by activity, but this response is delayed to the second day after acute exercise; repetitive exercise transiently depresses the expression of BDNF mRNA, so that the over-expression due to the previous day's exercise completely disappears 6 hours after the last exercise session.

[The best of nuclear cardiology and MRI in 2006]. / L'essentiel de 2006 en cardiologie nucléaire et IRM.

In this review, original clinical studies based on scintigraphy, PET or MRI imaging are reported, regarding the main areas of cardiology including ischemia and myocardial infarction, detection of coronary artery disease in various patients subsets, cardiomyopathy and heart failure. A special topic is dedicated to tracers and technical innovations.

Comprehensive complexity assessment as a key tool for the prediction of in-hospital mortality in heart failure of aged patients admitted to internal medicine wards.

Congestive heart failure (CHF) and cognitive impairment are both common problems in old age, associated with significant mortality, impaired quality of life and disability. This study evaluated patients with CHF, admitted to internal medicine and geriatric wards. We identified factors associated with a high risk of in-hospital mortality. Hospitalized CHF subjects with increased risk of in-hospital death present a clinical profile including: very old age, overt cognitive dysfunction, predisposition to falls, dependency, social-family problems, impairment in sphincter control and feeding ability, presence of bedsores, digoxin but not warfarin treatment, hypo-dysproteinemia and hypernatremia and mild renal impairment. We observed that patients admitted to our Internal Medicine Departments, in addition to CHF, present a high grade of complex therapeutic needs and that comorbidity, by itself, does not reflect complexity. Our data support the hypothesis that CHF has different patterns of severity and prognosis in young and in old or very old age groups.

Molecular characterisation of a Tuber borchii Smt3 gene.

Tbsmt3 gene from the ectomychorrizal fungus Tuber borchii was identified and sequenced. The Tbsmt3 gene encodes for a protein sharing significant amino acid homology with the yeast SMT3, a ubiquitin-like protein that is post-translationally attached to several proteins involved in many cellular processes. The comparison between the Tbsmt3 genomic and cDNA sequences established that the encoding sequence is interrupted by an intron of 312 bp. Southern blot analysis revealed only one copy of Tbsmt3 gene in the T. borchii genome. Tbsmt3 is expressed in all phases of T. borchii life cycle: mycelium, ectomycorrhiza and ascoma. However, the Tbsmt3 mRNA decreased during fruit body maturation.

[The best of nuclear cardiology and MRI in 2005]. / L'essentiel de 2005 en cardiologie nucléaire et IRM.

The literature concerning nuclear cardiology and cardiac MRI has been particularly rich in the fields of diagnosis, prognosis and therapeutic evaluation of coronary artery disease and cardiac failure. Almost 18 million 'conventional' myocardial scintigraphies (SPECT-single photon emission tomography, or TEM: tomography by monophotonic emission in French) are routinely performed worldwide each year. Nuclear cardiology represents the 3rd scientific domain of application for scintigraphy, after oncology and neurology. The advent of new conventional gamma cameras and PET (positron emission tomography) combined with CT will allow considerable improvement in the quality of investigation in obese or tri-truncal patients and women. We will limit ourselves to original clinical studies, based on scintigraphical techniques or magnetic resonance imaging, applied to the classic cardiological themes: myocardial infarction and ischaemia, cardiomyopathy and cardiac failure. We will also consider the new directions in nuclear cardiology regarding a new tracer and some innovative technology: rubidium-82 and TEP-CT.

Identification of differentially expressed cDNA clones in Tilia platyphyllos-Tuber borchii ectomycorrhizae using a differential screening approach.

No information is presently available on the molecular mechanisms that control the morphogenesis of the truffle, an ectomycorrhizal ascomycetous fungus of great economic interest not only for forestry and agronomy but also for the organoleptic properties of its hypogeous fruitbodies. A Tilia platyphyllos- Tuber borchii model system was used in order to identify genes induced or up-regulated during symbiosis, since their isolation is a prerequisite for the understanding of the molecular bases of mycorrhizal development and regulation. The strategy applied involved the construction of an ectomycorrhizal cDNA library and random selection of clones, followed by a differential screening procedure to analyse cDNA expression in uninfected roots, ectomycorrhizae and free-living mycelia. The results revealed that many genes - and more plant genes than fungal genes - are expressed at higher levels during the symbiotic phase. Several clones were also investigated in order to understand their biological function. This study represents the first attempt to extend our knowledge of the molecular mechanisms underlying the establishment of ectomycorrhiza in Tuber species.

Cloning and characterisation of a polyubiquitin gene from the ectomycorrhizal fungus Tuber borchii vittad.

Ubiquitin is a highly conserved 76-amino acid protein implicated in the function of quite different vital cellular processes. In the present study, we cloned and sequenced a polyubiquitin gene from Tuber borchii (Ubil) that is organised in four tandem repeats, with two C-terminal extension amino acids, serine and leucine. Two introns of 116 bp and 55 bp in length were detected in the first and second repeats, respectively. The Ubil gene is highly expressed in mycelium and is less expressed in the ripe fruiting body. Southern and Northern blot analyses revealed a second form of the ubiquitin gene.

Cloning, expression, and characterization of the hxk-1 Gene from the white truffle Tuber borchii vittad.: A first step toward understanding sugar metabolism.

Recent biochemical investigations of Tuber borchii Vittad. mycelium have demonstrated the presence of three distinct forms of hexokinase (HK(M1), HK(M2), and HKM3). In the investigation described here, a gene coding for hexokinase (hxk-1) from T. borchii was isolated and characterized. The hxk-1 gene is characterized by an ORF of 1494 nucleotides and codes for a polypeptide of 497 aa. The gene was overexpressed in Escherichia coli, and the recombinant protein was kinetically characterized. The K(cat) value for fructose is in agreement with the data reported for the hexokinase of Yarrowia lipolytica, the Km for ATP is not dependent on the sugar used, and the enzyme is not inhibited by trehalose 6-phosphate or glucose 6-phosphate. The biochemical characteristics confirm that this enzyme is a hexokinase, as suggested by the Pileup results, and it corresponds to the HKM1 isoform. This work represents the first characterization of the key enzyme of the glycolytic pathway and the related gene in a Tuber species.

One-step purification of a fully active hexahistidine-tagged human hexokinase type I overexpressed in Escherichia coli.

The conversion of glucose into glucose 6-phosphate (Glc 6-P)1 traps glucose in a chemical state in which it cannot leave the cell and hence commits glucose to metabolism. In human tissues there are at least three hexokinase isoenzymes responsible for hexose phosphorylation. These enzymes are constituted by a single polypeptide chain with a molecular weight of approximately 100 kDa. Among these isoenzymes, hexokinase type I is the most widely expressed in mammalian tissues and shows reversion of Glc 6-P inhibition by physiological levels of inorganic phosphate. In this work the hexokinase I from human brain was overexpressed in Escherichia coli, as a hexahistidine-tagged protein with the tag extending the C-terminal end. An average of 900 U per liter of culture was obtained. The expressed protein was one-step purified by metal chelate affinity chromatography performed in NTA-agarose column charged with Ni(2+) ions. In order to stabilize the enzymatic activity 0.5 M ammonium sulfate was added to elution buffer. The specific activity of purified hexokinase I was 67.8 U/mg. The recombinant enzyme shows kinetic properties in agreement with those described for the native enzyme, and thus it can be used for biophysical and biochemical investigation.

Regional myocardial metabolic rate of oxygen measured by O2-15 inhalation and positron emission tomography in patients with cardiomyopathy.

BACKGROUND: Positron emission tomography (PET) metabolic studies have investigated the pathways involved in fatty acid, glucose, and oxidative metabolism in cardiomyopathy and the impairments that occur in the damaged myocardium, but none have provided absolute quantitative variables. Recently, quantitative measurements of the metabolic rate of oxygen (MMRO2) and oxygen extraction fraction (OEF) using O2-15-labeled oxygen gas have been validated in animals and healthy volunteers. The purposes of the current study were to measure MMRO2 and OEF in cardiomyopathy with left ventricular (LV) dysfunction. METHODS: The authors selected 25 study participants: 16 patients (10 with ischemic and 6 with dilated) cardiomyopathy with LV dysfunction, and 9 healthy volunteers. As evaluated by echocardiography, LV ejection fraction (LVEF) was decreased in patients (35%+/-9% vs. 65%+/-5%, P<0.01). The PET protocol consisted of transmission, C O2-15 static, H2 O-15 dynamic, and O2-15 gas inhalation steady state scans. An entire myocardial region of interest was drawn to encompass the entire LV myocardium on three midventricular slices in each participant. RESULTS: Data showed in patients with dilated cardiomyopathy significant reductions of MMRO2 (0.051+/-0.02 ml x min(-1) x g(-1) vs. 0.108+/-0.02 ml x min(-1) x g(-1), p = 0.01) and OEF (0.55+/-0.15 vs. 0.71+/-0.08, P = 0.01) when compared with healthy volunteers. Furthermore, OEF decreased significantly in lateral and inferior walls. Significant correlations were observed among OEF and the rate-pressure product (RPP) (P = 0.02), LVEF (P<0.001), MMRO2 and RPP (P = 0.04), and LVEF (P = 0.05). In patients with ischemic cardiomyopathy, MMRO2 was significantly reduced (0.039+/-0.02 ml x min(-1) x g(-1) vs. 0.108+/-0.02 ml x min(-1) x g(-1); p = 0.005) but not OEF (0.63+/-0.2 vs. 0.71+/-0.08; P = NS), when compared with healthy volunteers. Significant correlations were observed among OEF and RPP (P = 0.03), LVEF (P = 0.002), MMRO2 and RPP (P<0.01), and LVEF (P = 0.03). CONCLUSIONS: These data show that O2-15 gas inhalation and PET allow myocardial MMRO2 and OEF to be measured in patients with cardiomyopathy.

Improvement of cardiac neuronal function after carvedilol treatment in dilated cardiomyopathy: a 123I-MIBG scintigraphic study.

UNLABELLED: Carvedilol can induce important clinical and hemodynamic improvements in patients with chronic heart failure resulting from severe left ventricular (LV) dysfunction. This study examines the impact of carvedilol on cardiac neuronal function using 123I-metaiodobenzylguanidine (MIBG) scintigraphy in dilated cardiomyopathy. METHODS: Twenty-two patients with chronic heart failure (19 men, 3 women; mean age, 54 y; age range, 34-64 y) assessed as New York Hospital Association (NYHA) class II or III and with initial resting radionuclide LV ejection fractions (LVEF) < 0.40 were enrolled in the study. Patients had long histories of symptomatic LV dysfunction despite optimal diuretics and angiotensin-converting enzyme inhibitor treatment. Over a 6-mo period, 50 mg/day carvedilol was administered to these patients. Planar 123I-MIBG scintigraphy provided measurements of cardiac neuronal uptake (as heart-to-mediastinum count activity ratio [HMR]), 4h after intravenous injection of 185 MBq MIBG. Hemodynamic, clinical, radionuclide LVEF and HMR data measured at the outset and after 6 mo of carvedilol were compared. RESULTS: Resting heart rate decreased from 81 +/- 13 to 71 +/- 9 bpm (P = 0.003). After carvedilol therapy NYHA functional classification for these patients improved from 2.6 +/- 0.5 to 2.3 +/-0.5 (P = 0.04), LVEF improved from 22% +/- 9% to 30% +/- 13% (P = 0.005), and HMR improved from 145% +/- 23% to 170% +/- 25% (P = 0.0001). CONCLUSION: Carvedilol induces improvements of clinical symptoms and cardiac neuronal and systolic functions in patients with dilated cardiomyopathy and chronic optimal treatment.

Impairment of cardiac neuronal function in childhood dilated cardiomyopathy: an 123I-MIBG scintigraphic study.

UNLABELLED: Abnormalities of norepinephrine uptake have been found to reflect impairment of cardiac adrenergic neuronal function in adults with heart failure. To our knowledge, no data on childhood dilated cardiomyopathy (DCM) are available. The aim of this study was to assess the cardiac neuronal function using 123I-metaiodobenzylguanidine (MIBG) scintigraphy in children with idiopathic DCM. METHODS: We studied 26 patients (mean age, 44+/-50 mo) with DCM and left ventricular dysfunction and 12 control subjects (mean age, 49+/-65 mo) with normal left ventricular function. All subjects underwent planar cardiac imaging after intravenous injection of 20-75 MBq 123I-MIBG. A static anterior view was acquired 4 h after injection. The heart-to-mediastinum count ratio was measured as described previously. RESULTS: On the basis of a reduction of the heart-to-mediastinum count ratio, cardiac neuronal uptake of 123I-MIBG was significantly decreased in patients with DCM compared with cardiac uptake in control subjects (172%+/-34% versus 277%+/-14%; P<0.0001). A significant correlation was found between left ventricular ejection fraction and 123I-MIBG cardiac uptake in patients with DCM (y = 2.5x + 113.3; r = 0.80; P<0.0001). CONCLUSION: Cardiac adrenergic neuronal function is impaired in children with idiopathic DCM. 1231-MIBG cardiac scintigraphy is a useful tool to assess cardiac neuronal function in childhood DCM.