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Reduced plasma adiponectin has been associated with abnormal lipid profile, reduced left ventricle (LV) function, and the extent of coronary atherosclerosis in coronary artery disease. The aim of this study was to assess these relationships in patients with dilated cardiomyopathy (DCM) without overt heart failure. Plasma adiponectin was measured in 55 DCM patients (age, 59 ± 12 years; male, 36; body mass index [BMI], 26.9 ± 0.49 kg/m²; LV ejection fraction, 39.8% ± 1.3%; New York Heart Association class I-II) and in 40 age- and BMI-matched healthy controls. In a subset of 25 patients, myocardial blood flow (MBF) was measured at rest and during intravenous dipyridamole (0.56 mg/kg in 4 minutes) by positron emission tomography and ¹³N-ammonia as a flow tracer. Adiponectin was 6.6 ± 0.34 μg/mL in controls and 10.9 ± 0.85 μg/mL in DCM patients (P < .001), where it was related inversely with BMI (P = .009) and directly with brain natriuretic peptide (P = .017), high-density lipoprotein (HDL) cholesterol (P = .002), and MBF dipyridamole (P = .020). Adiponectin lesser than median value in patients was associated with higher total to HDL cholesterol ratio (4.8 ± 0.24 vs 3.9 ± 0.18, P = .009) and lower MBF reserve (1.76 ± 0.16 vs 2.43 ± 0.19, P = .01). These results could suggest that down-regulation of the adiponectin levels and reduced HDL cholesterol have a key role in causing impaired coronary function and myocardial perfusion in DCM., (© 2011 Elsevier Inc. All rights reserved.)

C-type natriuretic peptide (CNP) plasma levels are extremely low and a pre-analytical phase is necessary to assay plasma CNP concentrations. Amino-terminal CNP (NT-proCNP) circulates at higher concentrations than CNP, allowing a direct assay and the use of smaller amounts of plasma. Aim of this study was to evaluate the analytical performance of a direct NT-proCNP assay and to measure its plasma levels in heart failure (CHF), diabetes and chirrosis patients. NT-proCNP and CNP were measured in 130 CHF, 19 patients with diabetes, 24 with hepatic cirrhosis and 73 controls. Plasma NT-proCNP was higher in all the clinical conditions studied (controls:45.5 ± 1.84 pg/ml, CHF:67.09 ± 7.36, diabetes:51.5 ± 5.75 cirrhosis:78.4 ± 19.9; p = 0.034, p = 0.04 controls vs. CHF and cirrhosis, respectively) and in CHF NT-proCNP concentration showed a significant increase as a function of clinical severity. By comparison of ROC curves, CNP assay resulted better associated with disease than NT-proCNP assay in all the different clinical conditions probably due to different release and clearance. The determination of NT-proCNP adds a piece of information to better understanding the molecular mechanisms at the basis of CNP action in different diseases. Due to its higher analytical feasibility, this determination could become widespread in clinical biochemistry laboratories and serve as a complementary marker of disease conditions., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Background: Redox aminothiols have been reported to modulate the activity of recombinant metalloproteinases (MMP). The aim of the present study was to investigate the effects of myocardial redox state on the activities of MMP-2 and -9 implicated in cardiac remodeling in end-stage heart failure patients supported by left ventricular assist device (LVAD)., Methods and Results: During heart transplant (HT) surgery, myocardial specimens (MS) from right ventricular walls and LV walls were obtained from 7 LVAD recipients (LVAD group, MS n=35) and from 7 stable HT candidates on medical therapy (MT group, MS n=35). Myocardial MMP-2 and -9 activities and expression, tissue inhibitor of MMP (TIMP)-1 and -4, transforming growth factor (TGF)-β1 and aminothiol concentrations were measured. MMP-2 and -9 activities were evaluated also by incubating MS with different amounts of reduced and oxidized glutathione (GSH). MMP-2 and -9 activities and expression were lower in the LVAD group, whereas myocardial TIMP-1 and -4 concentrations were comparable to those of MT patients. Higher GSH and TGF-β1 concentrations were found in LVAD-recipients. Only GSH concentrations were inversely related to MMP-2 and -9 activities. In vitro, GSH had an inhibitory effect on MMP-2 and -9 activities., Conclusions: LVAD recipients show reduced myocardial MMP-2 and -9 activities and expression when compared to medically treated patients. Changes of myocardial redox state, predominantly GSH-dependent, appear to modulate MMP-2 and -9 activities by an inhibitory effect dependent on thiol content. These data support a role of GSH cycle in modulating the extracellular matrix in end-stage heart failure patients supported by LVAD.

The cardiac resynchronization therapy (CRT), based on correction of electro-mechanical dyssynchrony by biventricular pacing in patients with severe chronic HF unresponsive to optimal medical treatment and left ventricular conduction disturbances, has been developed. The determination of plasma adrenomedullin (ADM) levels before implantation could provide important additional information to reduce the high percentage (30%) of patients not responding to treatment despite the use of increasingly sophisticated methods for selecting candidates. The case described illustrates the importance of basal ADM plasma levels in predicting the clinical and functional improvement after treatment with CRT.

Aim: the aim of this study was to determine neuropsychological performance (possibly predictive of academic difficulties) and its relationship with cognitive development and maternal education in healthy preterm children of preschool age and age-matched comparison children born at term., Method: a total of 35 infants who were born at less than 33 weeks' gestational age and who were free from major neurosensory disability (16 males, 19 females; mean gestational age 29.4wk, SD 2.2wk; mean birthweight 1257g, SD 327g) and 50 term-born comparison children (25 males, 25 females; mean birthweight 3459g, SD 585g) were assessed at 4 years of age. Cognition was measured using the Griffiths Mental Development scales while neuropsychological abilities (language, short-term memory, visual-motor and constructive spatial abilities, and visual processing) were assessed using standardized tests. Multivariable regression analysis was used to explore the effects of preterm birth and sociodemographic factors on cognition, and to adjust neuropsychological scores for cognitive level and maternal education., Results: the mean total Griffiths score was significantly lower in preterm than in term children (97.4 vs 103.4; p<0.001). Factors associated with higher Griffiths score were maternal university education (β=6.2; 95% confidence interval [CI] 0.7-11.7) and having older siblings or a twin (β=4.0; 95% CI 0.5-7.6). At neuropsychological assessment, preterm children scored significantly lower than term comparison children in all tests except lexical production (Boston Naming Test) and visual-processing accuracy. After adjustment for cognitive level and maternal education, differences remained statistically significant for verbal fluency (p<0.05) and comprehension, short-term memory, and spatial abilities (p<0.01)., Interpretation: neuropsychological follow-up is also recommended for healthy very preterm children to identify strengths and challenges before school entry, and to plan interventions aimed at maximizing academic success.

Background: Adiponectin, present in different multimeric forms in circulation, is increasingly used in clinical settings as a cardiometabolic marker. The development of several commercially available enzyme-linked immunosorbent assays (ELISA) has allowed for the widespread measurement of adiponectin in research as well as in clinical practice. The comparative performance of these assays is thus an issue of major relevance., Methods: The analytical performance of four different ELISAs (LINCO, B-Bridge, SPIbio and ALPCO) was evaluated. Samples from 102 cardiac patients and 40 healthy subjects were tested using LINCO and ALPCO. The latter is able to measure the concentrations of multimers. For the multimer assay, an error propagation study was performed. A subset of subjects was tested by all four ELISAs for comparison purposes., Results: Bland-Altman plots revealed differences between the results of the four ELISAs, mainly for ALPCO. However, a strong correlation between the different ELISAs (ALPCO, r=0.83; B-Bridge, r=0.98; SPIbio, r=0.91 vs. LINCO) was observed. Total adiponectin measured by LINCO showed a better association with cardiac disease [receiver-operating characteristic (ROC) curves] than total and high molecular weight adiponectin measured by ALPCO., Conclusions: The results of this study contribute to a better understanding of the methodological features of the several ELISAs, and help in the evaluation and comparison of the relative results.

In patients with dilated cardiomyopathy (DCM) abnormal myocardial blood flow (MBF) has been associated with coronary microvascular dysfunction. The aim of this study was to test the hypothesis that osteopontin (OPN) plasma levels could be associated with the activation of the renin-aldosterone system (RAS) in these patients and be involved in mediating myocardial and coronary damage. In 66 patients with idiopathic left ventricular dysfunction of variable severity the plasma levels of OPN were correlated with biomarkers of systemic metabolism, RAS activation, myocardial dysfunction and with clinical indexes of left ventricle (LV) function and perfusion obtained by 2D-echocardiography and PET. As compared to controls, patients showed a significant increase of inflammatory markers (OPN: 508+/-30.8ng/ml vs. 426.9+/-16.4, p<0.05 and interleukin (IL)-6: 1.71+/-0.29pg/ml vs. 0.38+/-0.03pg/ml, p<0.001) and of indexes of cardiac damage. OPN levels were significantly correlated with the extent of microvascular dysfunction (MBF at rest: p=0.01; during dipyridamole: p=0.0003) and with plasma renin activity (PRA) (r=0.26, p=0.04). Both in patients with milder or more severe LV dysfunction lower MBF values were associated with higher OPN levels and PRA. These results suggest a interdependent role of RAS and vascular inflammation in cardiomyopathy., (2009 Elsevier Ltd. All rights reserved.)

We evaluated language development in deaf Italian preschoolers with hearing parents, taking into account the duration of formal language experience (i.e., the time elapsed since wearing a hearing aid and beginning language education) and different methods of language education. Twenty deaf children were matched with 20 hearing children for age and with another 20 hearing children for duration of experience. Deaf children showed a significant delay in both vocabulary and grammar when compared to same-age hearing children yet a similar development compared to hearing children matched for duration of formal language experience. The delay in linguistic development could be attributable to shorter formal language experience and not to deafness itself. Deaf children exposed to spoken language accompanied by signs tended to understand and produce more words than children exposed only to spoken language. We suggest that deaf children be evaluated based on their linguistic experience and cognitive and communicative potential.

In humans, the Apelin gene is located on chromosome Xq25-26.1 and it encodes a 77-aminoacid prepropeptide. Considerable sequence homology exists across different mammalian species. Apelin is emerging as an important regulator of cardiovascular homeostasis but, at present, few data from humans are available and further studies are necessary to better define its role in cardiovascular pathophysiology. The role and function of Apelin in cardiovascular system could be reliably investigated in experimental models devoid of confounding effects reflecting only the natural history of the disease. The pig constitutes a model largely used in experimental pathology where it has a central role in "in vivo" clinical settings. Sus Scrofa genoma is not completely sequenced and Apelin gene is still lacking. Aim of this study was to sequence the Apelin in Sus Scrofa for future applications to molecular biology studies. Using the guanidinium thyocyanate-phenol-chloroform method, we extracted total RNA from samples obtained from heart of mouse and from atrium and ventricle of normal pigs. Pig Apelin mRNA was sequenced using polymerase chain reaction primers designed from mouse consensus sequences. A partial sequence of Sus Scrofa Apelin mRNA, 1-201 pb, was submitted to GenBank (accession number FJ362603). The bands obtained from pig cardiac tissue shared a 99% sequence identity with Mus musculus and 90% with Rattus norvegicus. The knowledge of Apelin sequence can be an useful starting point for future studies devoted to better understand the possible alterations of Apelin mRNA expression in different cardiac diseases.

High-frequency pacing of the left ventricle (LV) free wall causes a dyssynchronous pattern of contraction that leads to progressive heart failure (HF) with pronounced differences in regional contractility. Aim of this study was to evaluate possible changes in brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) mRNA expression in the anterior/anterior lateral region (pacing site, PS) as compared to the infero-septal region (opposite site, OS) and to explore possible association between the contractiling pattern and biomarker expression. Cardiac tissue was collected from minipigs with pacing-induced HF (n=8) and without (control, n=6). The samples were selectively harvested from the anterior left ventricular (LV) wall, PS, and from an area remote to the pacing-site, OS. BNP and CNP mRNA expression was evaluated by semi-quantitative polymerase chain reaction (PCR). A significant difference in BNP expression was found in the PS between HF animals and controls (BNP/GAPDH: 0.65+/-0.11 vs. 0.35+/-0.04, p=0.02), but not in the OS (BNP/GAPDH: 0.36+/-0.05, ns vs. controls). CNP expression was not different compared to controls, although higher levels were observed in the PS and in the OS with respect to the controls (CNP/GAPDH: controls 0.089+/-0.036, PS 0.289+/-0.23, OS 0.54+/-0.16). This finding was in tune with an increase of CNP tissue concentration (controls: 0.69+/-0.13; PS=1.56+/-0.19; OS=1.70+/-0.42 pg/mg protein; p=0.039 controls vs. OS). Higher BNP mRNA expression in the PS is consistent with a reduction in contractile function in this region, while higher CNP mRNA expression in the OS suggests the presence of concomitant endothelial dysfunction in the remote region.

Background: Overexpression of heat shock proteins (Hsps) is associated to myocardial protection and it has been suggested that they could be a marker of cardiac preservation in conditions such as extracorporeal circulation. Aim of this study was to evaluate if cardioplegic arrest can modify the expression of Hsps in the heart and if this alteration is associated to cardiac preservation., Method: The levels of Hsp 27, Hsp 60, and both the constitutive and the inducible form of Hsp 70 were measured in the cardiac tissue from right atrium of pediatric patients before and after aortic cross-clamping (ACC) during cardiopulmonary bypass surgery for correction of congenital heart disease (n=20). The quantitative evaluation of Hsps was made by Western blotting analysis after tissue extraction and protein separation. Hsp 72 mRNA expression was also evaluated in pre- and post-ACC samples of eight subjects by semiquantitative RT-PCR. Peripheral levels of Troponin I, Myoglobin, LDH, CK, CK-MB were measured in basal conditions and at 12 and 24h after cardiosurgery as markers of heart damage., Results: The cardioplegic arrest did not significantly modify the mean levels of all the Hsps measured. Hsp 72 levels increased after cardioplegia in the 40% of the patients and all Hsps in the 28% of subjects. The patients whose levels of Hsps are increased after cardioplegia are associated with lower post-surgery concentrations of all the markers of cardiac injury., Conclusions: This observation suggests a relationship between the increase of Hsps and the reduction of cardiac injury.

1. Lactase, sucrase, maltase, trehalase and alkaline phosphatase activities of rat proximal jejunum were measured in 3, 6, 9, 12, 18 and 24-month-old rats fed with diets differing in their fatty acid composition. 2. A drop of 47-53% of the specific enzyme activity was observed with disaccharidases against a decrease of 71% for alkaline phosphatase in the 24-month-old rats compared to the 3-month-old rats. 3. Changes in dietary fatty acid composition, either in the saturated or monounsaturated ratio, or in the polyunsaturated fatty acid composition, did not significantly interfere with this aging effect.

1-14C linoleic acid intestinal absorption and simultaneous biochemical events were followed up on rats under vascular perfusion and on main mesenteric lymphatic duct fistulated rats. 1-14C linoleic acid was introduced in the duodenum alone in doses from 1.2 to 90 mumol or in the presence of oleic acid and monopalmitin (30/30/30 mumol/mumol/mumol). Mesenteric portal venous blood and chyle, respectively, were collected continuously for 1 and 6 h after the infusions. Blood-labeled lipid recovery varied from 4.7 to 2.2% of the 14C linoleic acid infused as the 14C linoleic acid dose infused increased, and dropped to 1.8% with the mixed lipid infusate. Lymph-labeled lipid recovery increased from 25.7 to 31.8% of the 14C linoleic acid infused as the dose infused increased, and rose to 48.1% with the mixed lipid infusate. The oxidation of 1-14C linoleic acid remained low: 0.8-3% of the infused radioactivity. A desaturation and elongation of 14C linoleic acid into 14C arachidonic acid was detected and discussed. We can conclude that the linoleic lymph absorption pathway remained preferential in our experimental conditions, simultaneous to a low rate of oxidation and an eventual ability for the enterocyte to convert this essential fatty acid arachidonic acid.

The biochemical and ultrastructural aspects of intestinall fat absorption and chylomicron formation have been studied in rats treated with actidione-cycloheximide. Rats were prepared with a catheter in the main mesenteric lymph duct. The next day, primor rapeseed oil containing 14C triolein was infused in the duodenum of control rats and actidione-cycloheximide treated rats four hours before experiments. In each case, the quantity of lipids absorbed was determined after six hours, and the lipids present in the mucosa were studied as were those present in the lymph collected throughout the absorption period. Furthermore, the ultrastructural study of the mucosa and the morphological study of the lymph lipoprotein particles were carried out using electron microscopy. Disappearance of intraluminal lipids and mucosal lipids appeared slightly impaired after actidione-cycloheximide treatment, but only a very small proportion of the infused lipids were recovered in the intestinal lymph (0.05% compared to 3% in control rats, Table I). The amount of esterified lipids found in the mucosa, after six hours, was clearly lower (49%) when compared with those of control rats (79%) (Table II). This suggests that, following inhibition of protein synthesis by actidione-cycloheximide, lymph fat absorption was much impaired. An alteration in cellular structure appeared in microscopic observations of treated rat enterocytes (Fig. 2 and 3) compared with the control (Fig. 1). Many whorl formations and autophagic vacuoles were included in these cytotoxic effects during lipid absorption. Treated animal enterocyte lipid particles did not have the organized structure of those in the controls. While lymph particles of control rats revealed a regular progression in size (Fig. 4A, B), lymph particles of treated-rats showed a few large chylomicrons (equal or far larger than in the controls) among small particles (Fig. 4 C, D, E); furthermore, evidence of coalescence might be observed (Fig. 4 F). All these observations emphasize the prevailing role of the different protein structures of the enterocytes during lipid intestinal absorption, particularly for chylomicron organization after long-chain fatty acid uptake.

Studies of the development of sentence comprehension strategies in English have indicated that, at first, children tend to rely on pragmatic and semantic strategies, whereas, later on, they rely primarily on word order to determine the basic grammatical relations. However, before making strong conclusions regarding the role of semantics in comprehension, it is necessary to distinguish between (1) extragrammatical knowledge of world events and (2) abstract semantic distinctions that may be an integral part of the parsing process. An earlier study of American and Italian adults indicated that use of such abstract semantic strategies may be a core strategy for parsing in Italian. In the present study, we compared sentence interpretation in American and Italian children between the ages of 2 and 5. From the earliest stages, children showed sensitivity to the relative information value of the various cues in their native language; Italians relied primarily on semantic cues, whereas American children relied on word order. In general, the data did not support claims regarding the existence of universal hypotheses about language structure. There was also evidence that the failure of these young children to make full use of certain interpretive cues resulted from their inability to appreciate the discourse functions of these cues.

Rats were fed on a 20% fat diet; 30 min. or 2 hrs later intestinal jejunal cells were fixed in situ; samples were taken and processed for subsequent study by electron microscopy. Chylomicron secretion was followed from the enterocyte to the lymphatic capillary lumen. Chylomicrons entered the lamina propria through breaks in the basal membrane at intercellular space level. Their penetration into the lymphatic capillary lumen occurred through the open junctions of the endothelial cells normally kept closed by desmosomal formations.