Closure of an unusual morphology patent ductus arteriosus with a covered stent in a dog.

Patent ductus arteriosus (PDA) with unusual morphology was diagnosed in a 3-year-old German shepherd presented for exercise intolerance. Two interventional PDA closure procedures with various Amplatzer devices and one surgical ligation were attempted to close the PDA. However, PDA closure failed with both methods due to the unusual morphology and the severe enlarged ductus ampulla. Therefore, the patient underwent an aortic covered stent placement, and successful closure of the PDA was achieved. Eight months after the procedure, the dog was free of clinical signs and no residual flow was identified through the PDA on transthoracic echocardiography. Moreover, computed tomography angiography demonstrated correct stent positioning, with no evidence of thrombus formation, damage to the stent, nor any aortic wall abnormalities. Aortic covered stent placement is a viable option in dogs with unusual PDA morphology.

Efficacy and safety of ezetimibe/simvastatin association on non-diabetic and diabetic patients with polygenic hypercholesterolemia or combined hyperlipidemia and previously intolerant to standard statin treatment.

BACKGROUND AND OBJECTIVE: One of the problems associated with reaching the low-density lipoprotein cholesterol (LDL-C) target during statin treatment is the emergence of laboratory or clinical side effects. The aim of our study was to evaluate the prevalence of statin-associated adverse events in diabetic and non-diabetic patients affected by polygenic hypercholesterolemia or combined hyperlipidemia and the efficacy and tolerability of treatment with ezetimibe/simvastatin 10/10 mg/day on the same subjects experiencing the adverse events. METHODS: Consecutively enrollment of patients affected by polygenic hypercholesterolemia or combined hyperlipidemia with or without type 2 diabetes mellitus. Each Centre used any of the available statins on the basis of current clinical judgement and monitored enrolled patients for adverse events during the following 2 years. Those patients with moderate adverse events suspended the current statin therapy for 1 month (washout period), and then were shifted to treatment with ezetimibe/simvastatin 10/10 mg/day and again monitored for adverse events in the following 6 months. We assessed body mass index, glycated haemoglobin, fasting plasma glucose, total cholesterol, LDL-C, high-density lipoprotein cholesterol, triglycerides, alanine aminotransferase, aspartate aminotransferase, creatinine phosphokinase and monitored adverse events such as asthenia and myalgia. RESULTS AND DISCUSSION: All 1170 Caucasian patients affected by polygenic hypercholesterolemia obtained a significant reduction in LDL-C during the observation period (P < 0*05), while those with combined hyperlipidemia also showed a reduction in TG plasma level (P < 0*05) and a significant increase in HDL-C (P < 0*05). Patients affected by polygenic hypercholesterolemia experiencing adverse event under statin treatment obtained a significantly lower reduction than those tolerating the treatment (P < 0*001). The prevalence of adverse events under statin treatment was 4*9% in non-diabetic patients with polygenic hypercholesterolemia, 8*6% in those with combined hyperlipidemia, 7*1% in diabetic patients with polygenic hypercholesterolemia and 7*6% in those with combined hyperlipidemia. Six months after the shift to treatment with ezetimibe/simvastatin 10/10 mg, all patients experienced a significant improvement in LDL-C, TG and HDL-C plasma level. No adverse event was registered during the ezetimibe/simvastatin 10/10 mg treatment period. It seems that previous side effects observed with statins did not re-appear with the administration of ezetimibe/simvastatin 10/10 mg/day. CONCLUSIONS: The efficacy and adverse effect profile of the ezetimibe and simvastatin combination appear to be good for both diabetic and nondiabetic patients, and in both conditions.

European official control of food: Determination of histamine in fish products by a HPLC-UV-DAD method.

The evaluation of histamine content in fish and fishery products, responsible for scombroid poisoning, is essential to guarantee the safety of food. EU regulation requires validated analytical methods to ensure the verification of compliance with food law in official control activity. To this aim a previous gradient RP-HPLC method with DAD detection was modified and validated, according to international guidelines. The reliability of results was tested by analysing fish reference materials within the participation in European proficiency tests. The method has been used for the analysis of real samples consisting of several fish-based products with considerable differences in matrix composition. This characteristic is of great relevance to be able of apply the method in the field of official control.

Double-blind comparison of perindopril and captopril in hypertension. Effects on left ventricular morphology and function.

Using digitized M-mode echocardiograms, we compared, in a double-blind study, the effects of 4 to 8 mg perindopril given once daily and 25 to 50 mg captopril given twice daily on the left ventricle (LV) in 20 hypertensive patients. Both treatments significantly (P less than .001) lowered blood pressure, reducing systemic vascular resistances. After 3 months both drugs induced a comparable percentage of reduction in LV mass, with an increase in the peak rate of LV relaxation and no changes in the peak rate of LV contraction. Our results demonstrate that perindopril once daily is an effective antihypertensive agent; it is also able, like captopril, to induce regression of LV hypertrophy, with improvement in diastolic performance and no deterioration in ventricular systolic function.

Improved methods for the preparation of high molecular weight DNA from large and small scale cultures of filamentous fungi.

Improved methods are described for the isolation of pure, high molecular weight DNA from small and large scale cultures of filamentous fungi. The methods depend on the extraction of DNA under conditions which prevent nuclease activity and contamination by carbohydrate. The small scale method depends on enzymatic digestion of the wall whereas the large scale method uses partial damage followed by autolysis. High yields of DNA are obtained by both methods and the DNA is suitable for restriction analysis. Southern Blotting, RFLP analysis, dot blotting and the production of gene libraries. The small scale method can be used for the simultaneous analysis of multiple cultures.

Oral fish cartilage polysaccharides in the treatment of photoageing: biophysical findings.

The topically applied cosmetic products can be helpful in improving the aged skin condition. The present study shows how oral fish-cartilage food supplementation can be helpful in improving the treatment of ageing skin. A total of 30 healthy women with signs of skin ageing were studied. Fifteen of the women were treated with a food supplement based on polysaccharides derived from the fish cartilage and a natural mix of antioxidants for 2 months and the other 15 with a placebo. Clinical evaluation and biophysical parameters related to skin function and wrinkle severity, such as silicone replica, skin thickness, mechanical properties, skin colour and capacitance, were measured. The results showed statistically significant changes in the active-treated group in comparison to the placebo. In particular, dermal thickness (treatment: from 1.13 to 1.23 mm; P < 0.001), skin wrinkling (treatment: from 9.5 to 3.5 R(a); P< 0.002), skin colour (treatment: brighter and less pigmented; P < 0.02) and viscoelasticity (treatment: from 0.70 to 0.97%; P < 0.02) showed considerable improvement. Most of these parameters are related to changes occurring within the dermal matrix, which is improved after the treatment, whereas most of the topically applied cosmetic products have a short-term effect on superficial structures. A combination treatment (oral and topical) can be more effective in reducing the signs of skin-ageing.

[Recent findings on the phenotype and function of T-lymphocytes in the human colostrum]. / Recenti acquisizioni sul fenotipo e la funzione dei T linfociti presenti nel colostro umano.

Recent studies in our laboratory have demonstrated that the great majority of human colostral T cells display the phenotypic and functional characteristics of memory T lymphocytes, e.g. were able to proliferate in response to anti-CD3 and anti-CD2 monoclonal antibodies, and to a lesser extent, to the lectin mitogen phytohaemagglutinin. In addition, their production of interferon-gamma after anti-CD3 and anti-CD2 stimuli was similar to that calculated in autologous blood lymphocyte cultures. More interestingly, the proportion of T lymphocytes bearing the gamma/delta T-cell receptor was found to be significantly higher in the mammary secretion than in autologous and heterologous blood samples. Furthermore, these cells were mostly delta-TCS-1+, thereby suggesting that they are actively motile cells capable of migrating from lymphoid to extra-lymphoid body tissues. The fact that the phenotypic pattern of colostral gamma/delta T cells is similar, if not identical, to that of the intestinal intraepithelial counterpart suggests that these cells might originate in the gut-associated lymphoid system and home selectively to the mammary gland late in pregnancy and throughout lactation. However, additional studies are needed to confirm whether milk T lymphocytes are actively involved in the adoptive lactation transmission of cellular immunity to the suckling infant.

Assessment of jugular endovascular malformations in chronic cerebrospinal venous insufficiency: colour-Doppler scanning and catheter venography compared with intravascular ultrasound.

OBJECTIVES: Chronic cerebrospinal venous insufficiency (CCSVI) is a malformative condition characterized by several anomalies of the azygos and/or internal jugular veins (IJVs). Recommended diagnosis of CCSVI is performed with colour-Doppler (CD) sonography. Though catheter venography (CV) is considered as the gold standard for determining vascular anatomy, its uniplanar point of view does not allow an overall evaluation of endoluminal structures. This limit could be addressed by intravascular ultrasound (IVUS). The aim of this report is to evaluate, in patients with multiple sclerosis (MS), the accuracy of CD sonography and CV versus IVUS in estimating the diameter and the cross-sectional area (CSA) of the IJVs and in detecting jugular endoluminal malformations (JEM). METHOD: Forty-five MS patients with CCSVI, diagnosed by CD sonography, were submitted to CV during IJVs angioplasty. Twenty-five subjects were also examined with IVUS. The IJVs maximum diameter (MAXD) and CSA were estimated. CD and CV data were compared with IVUS data with the Bland-Altman method. RESULTS: The mean difference in IJV MAXD recorded by CD and IVUS was -0.5 mm. The mean difference in IJV MAXD recorded by CV and IVUS was 3.36 mm. The mean difference in IJV CSA recorded by CD and IVUS was -11.2 mm(2). JEM recorded by IVUS were detected by CD sonography and CV with 88% and 32% accuracy, respectively. CONCLUSIONS: CV was significantly inferior to CD sonography and IVUS in detecting JEM. Differences between IVUS and CD sonography in detecting JEM and in quantifying jugular diameters were not significant. The IJV CSA was underestimated by CD sonography compared with IVUS. CD sonography was proven to be important in the anatomical characterization of CCSVI, providing useful information for correct intravascular treatment.

Epidemiological and virological characterization of a large community-wide outbreak of hepatitis A in southern Italy.

A large outbreak of hepatitis A virus (HAV) infection occurred in 2004 in Campania, a region of southern Italy, with 882 cases reported between 1 January and 1 August. The local public health authorities and the Italian National Institute of Health carried out investigations in order to characterize the agent, identify the source of infection and the route of transmission, and implement appropriate control measures. A web-based reporting system enhanced the flow of information between public health authorities, providing real-time epidemic curves and frequency distributions. The same 1B HAV genotype was found in 90% of sera from a subset of patients with acute disease, suggesting a local common source. A case-control study in the municipality with the highest attack rate showed that raw seafood consumption, in particular if illegally sold in water, was strongly associated with HAV illness. Samples of seafood systematically collected from retailers were found contaminated by HAV.

Temporal pattern of appearance and distribution of cholecystokinin-like peptides during development in Xenopus laevis.

Cholecystokinin (CCK) radioimmunoassay and immunocytochemistry were used to localize the expression of CCK-like peptides in the gut, brain, and skin of Xenopus laevis during development from fertilization through metamorphosis to the adult form. CCK-like peptides were first detected in the gut shortly before food was first observed in the lumen of the intestine. This CCK-like immunoreactivity (CCK-li) was localized in endocrine cells in the duodenal mucosa. Gut CCK-li decreased during metamorphosis coincident with gut histolysis. After metamorphosis, gut CCK-li appeared in the newly organized gastric antrum at Nieuwkoop-Faber stage 62-63 and reappeared in the reconstituted duodenum at stage 63-64. In both cases, CCK-li was localized in endocrine cells. In both the gastric antrum and the duodenum, the first appearance of CCK-li preceded the functional activity of its target tissues. CCK-li increased in the brain during brain primary differentiation and again during metamorphic climax coincidentally with the differentiation of the hypothalamo-hypophysial system. Brain CCK-li then declined during the transition from larva to adult. We conclude that the first presence of food in the lumen of the larval gut does not induce the appearance of CCK-li and that CCK may play a role in regulating the development of the gastrointestinal tract and brain in X. laevis.

Human milk lymphocytes bearing the gamma/delta T-cell receptor are mostly delta TCS1-positive cells.

Our laboratories previously reported that the proportion of lymphocytes bearing the gamma/delta T-cell receptor (TcR) is twofold greater in human milk lymphocyte suspensions than in autologous and heterologous blood samples. The present investigation shows that gamma/delta colostral T cells preferentially express non-covalently bound gamma/delta chains, whereas only a few display the disulphide-linked form of the TcR. This contrasts with the picture in the blood, where the distribution of these two nonoverlapping subpopulations is proportionally inverse. These findings are further evidence that the colonization of the mammary gland during lactation by immune system cells is the result of a selective homing process.

T-lymphocyte activation pathways in alcoholic liver disease.

Immune system derangement in cirrhotic patients with evidence of malnutrition is a well-recognized characteristic of chronic alcohol abuse. However, in vitro studies on cellular immune function performed with lectin mitogens have produced conflicting results. The recent development of more accurate immunological techniques for studying lymphocyte transformation, that use monoclonal antibodies directed against surface structures (CD3 and CD2) involved in antigen recognition, as well in adhesion functions, prompted us to study discrete in vitro T-cell hypo-responsiveness in a series of alcoholic liver disease (ALD) patients with no evidence of malnutrition or hepatic cirrhosis. The results indicated that the CD2 pathway is markedly defective in ALD T lymphocytes, accompanied by reduced interleukin-2 (IL-2) receptor expression upon in vitro activation. This defect cannot be reversed by the addition of recombinant IL-2 (rIL-2) or rIL-1. Faulty intracellular signal transduction by protein kinase C (PKC) and defective intracellular Ca2+ mobilization may be responsible for the CD2 pathway impairment. The addition of small amounts of phorbol 12-myristate, 13-acetate, but not Ca2+ ionophore A23187, is able to overcome the defect, thereby suggesting a direct PKC involvement. The hypothesis of a direct ethanol effect on transmembrane signal transduction systems is suggested by the demonstration of an expansion of circulating virgin (naive) T cells (CD3+/UCHL1-low) that binds tyrosine phosphatase (CD45RA antigen) on their surface.

Is the shortening of the QTc interval in Q-wave leads showing ST segment shift during exercise testing a new ECG marker of myocardial ischemia and viability in patients with previous anterior myocardial infarction?

PURPOSE: To evaluate whether the shortening of the QTc-interval, measured in Q-wave leads showing ST segment elevation during exercise testing may be a marker of stress-induced transmural ischemia (and indirectly of myocardial viability) in the infarct zone in patients with prior Q-wave anterior myocardial infarction. METHODS: We evaluated 15 consecutive patients (Group A) with previous anterior myocardial infarction presenting these peculiarities: 1) ST segment elevation over Q waves during exercise testing; 2) critical (> 75%) stenosis of LAD; 3) evidence by echocardiography and stress-redistribution-reinjection 201thallium myocardial scintigraphy (SRR201TIMS) of viable myocardium in the infarct zone (akinetic segments with normal echo-reflectivity plus > 7 mm end-diastolic wall thickness and significant 201thallium redistribution after reinjection). The study control group (Group B) consisted of 15 patients with previous myocardial infarction, critical stenosis of LAD and evidence of scarring by imaging techniques (increased echo-reflectivity associated with an end-diastolic wall thickness < 6 mm and no 201thallium redistribution in infarcted areas). The QTc interval was measured at rest and at peak stress in all leads, and particularly in infarct-related leads showing ST-T changes, and the lead-by-lead fractional difference percentage between the QTc intervals (delta QTc) was calculated. The delta QTc was measured again during exercise testing in 11 patients from Group A (Group A1) who showed significant contractility recovery three months after complete myocardial revascularization. A delta QTc shortening < -10% was considered "significant". RESULTS: In 14/15 patients from Group A, a significant delta QTc shortening was measured, while in 14/15 patients from Group B no significant delta QTc shortening was detected (sensitivity = 93.3%; specificity = 93.3%) (p < 0.0001). The mean delta QTc in Group A was -18.1 +/- 8.5%; the mean delta QTc in Group B was -4.2 +/- 7.8% (p < 0.0001). No patient from Group A1 showed a significant delta QTc shortening in Q-wave leads (mean delta QTc group A1 = +6.9 +/- 14.8%). CONCLUSIONS: delta QTc shortening in infarct-related leads during exercise testing is a simple ECG marker of transmural ischemia and, indirectly, of myocardial-viability. This sign is no more evident after myocardial revascularization and may be useful in identifying "hibernating-myocardium".

Lymphocytes bearing the T cell receptor gamma delta in human breast milk.

Lymphocytes bearing the T cell receptor gamma delta (TCR-gamma delta) were searched for in human early milk lymphocyte suspensions by two colour cytofluorimetric analysis. It was found that the proportion of TCR-gamma delta+ cells was twofold greater in colostrum than in either autologous or heterologous blood samples. Additional studies are needed to determine whether this particular subset of lymphocytes is involved in the lactation transmission of cellular immunity.

Human breast milk T lymphocytes display the phenotype and functional characteristics of memory T cells.

Naive (unsensitized) and memory (antigen-primed) T cells can be phenotypically distinguished on the basis of the high or low intensity with which they express a number of immunologically relevant lymphocyte membrane antigens, including CD45R, CDw29, UCHL1, LFA-1, LFA-3, CD2 and Pgp-1. Here we report that in contrast to the two major T cell subsets found in the blood, milk T lymphocytes are almost exclusively composed of the one which exhibits the CD45Rlow, CDw29, UCHL1, LFA-1high memory T cell phenotype. In addition, while milk and autologous blood cells expressed similar levels of CD3 surface antigens, CD2 and ICAM-1 expression was approximately twofold greater on the milk T lymphocytes. This agrees with the finding that whereas colostrum T cells respond poorly to PHA, they proliferate and produce interferon-gamma normally when stimulated with either the anti-CD3 or anti-CD2 monoclonal antibodies. The selective colonization of the mammary gland during lactation by a population of T lymphocytes which displays the phenotype and functional characteristics of memory T cells may be one of the mechanisms whereby the suckling infant benefits form its mother's immunological experience.

Influence of age and sex on left ventricular anatomy and function in normals.

Using digitized M-mode echograms, we evaluated the influence of sex on age-related changes of left ventricular (LV) anatomy and function in a normal population (75 males and 75 females, subdivided in age groups for each decade from 20 to 70 years). Aging is accompanied with an increase in septal and wall thickness in both males and females and in LV diameter only in males, with a progressive increase of LV mass more pronounced in males than in females. As regards LV function we found a progressive slowing of relaxation in females and of both contraction and relaxation in males, not related to changes in LV mass.

Lymphocytes bearing the gamma delta T cell receptor in acute Brucella melitensis infection.

A phenotypical analysis carried out by indirect immunofluorescence and two-color cytofluorometry showed that the number of lymphocytes bearing the gamma delta T cell receptor (TcR) heterodimer was dramatically increased in the blood of six children with Brucella melitensis infection. Most in vivo expanded gamma delta T cells reacted with a monoclonal antibody which identifies V delta 2 gene products and a significant proportion expressed CD25 and HLA-DR activation antigens. In addition, whereas only a few gamma delta T lymphocytes were CD8+, nearly all were CD4-. Highly enriched populations of both alpha beta and gamma delta T cells were obtained by negative immunoselection from three subjects with brucellosis sampled during convalescence. Despite the different form of their TcR, the proliferation of these two major T cell subsets in response to a mitogenic anti-CD3 monoclonal reagent (OKT3) was optimal. In contrast, alpha beta, but not gamma delta, T lymphocytes proliferated vigorously in response to the antigenic stimulus elicited by heat-killed Brucella. Further studies are, therefore, needed to determine whether the selective expansion of the gamma delta T cell subpopulation observed during the clinical course of the infection is driven by antigenic determinant(s) borne by the pathogen in vivo or is due to host-derived stimuli, such as autologous heat-shock proteins expressed on the surface of the infected cells.

T-cell response to phorbol ester PMA and calcium ionophore A23187 in Down's syndrome.

The proliferative response of purified T cells to anti-CD2 monoclonal antibodies (T112 plus T113) was found to be markedly reduced in 12 subjects with Down's syndrome (DS). The addition of phorbol ester PMA, which activates Ca2+/phospholipid-dependent enzyme protein kinase C, or calcium ionophore A23187, which increases intracytosolic free Ca2+ concentration, enhanced, but did not normalize, the defective anti-CD2-mediated T-cell mitogenesis. In contrast, the proliferation of resting lymphocytes from trisomic patients was comparable to that of the control cells when PMA and A23187 were used as co-blastogenic reagents. Because PMA and A23187 together bypass the early activation pathways and promote T-cell growth through the direct induction of membrane interleukin 2 (IL-2) receptor expression and IL-2 synthesis and secretion, it could reasonably be hypothesized that the faulty DS T-cell activation induced by antigen or mitogen is due to a deranged transmembrane signal transduction, rather than a defect in the later intracellular events.

[Factors influencing left ventricular function in arterial hypertension]. / Fattori che influenzano la funzione ventricolare sinistra nella ipertensione arteriosa.

Using digitized M-mode echograms we evaluated the role of preload, afterload, inotropic state and left ventricular (LV) mass on LV systolic and diastolic function in 2 groups of hypertensive patients: Group 1: 25 subjects (18 men, mean age 48 +/- 6 years) with normal LV mass (less than 230 g); Group 2: 25 subjects (20 men, mean age 50 +/- 8 years) with LV hypertrophy (wall hypertrophy with normal LV diameter). As control group, we evaluated 50 normal subjects, matched for age, sex and body surface area with hypertensives. LV mass was significantly (p less than 0.001) higher as respect to normals also in hypertensives with normal LV mass; indexes of LV systolic and diastolic function were similar in normals and in hypertensives with normal LV mass and significantly lower in subjects with LV hypertrophy. The end-systolic wall stress was not significantly different in the 2 groups of hypertensives. We evaluated the relative role of preload (end-diastolic LV diameter), afterload (end-systolic wall stress) inotropic state (systolic arterial pressure/end-systolic LV diameter) and LV mass on LV systolic and diastolic function using multiple regression analysis. As regards LV systolic function, the major determinant was the systolic pressure/end-systolic diameter ratio in normals, the end-systolic stress in hypertensives. As regards LV diastolic function, the major determinant was end-systolic stress in normals and hypertensives with normal LV mass, LV mass in hypertensives with myocardial hypertrophy. Preload seems not to influence LV function in normals and in hypertensives with normal LV diastolic diameter. The major determinant of LV systolic function is the inotropic state in normals and the afterload in hypertensives.(ABSTRACT TRUNCATED AT 250 WORDS)

Spermidine labels proteins during sea urchin embryogenesis.

We have previously described the presence of a protein containing intact, covalently bound spermidine during very early embryogenesis of the sea urchin (Strongylocentrotus purpuratus). Proteins containing other polyamine metabolites also appear as embryogenesis proceeds. These proteins which contain label derived from exogenous radioactive spermidine show a characteristic pattern which changes during the course of embryonic development. We document for the first time that hypusine, the polyamine metabolite which is a characteristic component of the eukaryotic protein translation initiation factor eIF-4D, is present in more than one species of macromolecule. In addition, N1-acetylspermidine has also been identified as a significant intracellular metabolite of spermidine during embryogenesis.