Smoking and hypertension: Effect of adenosine deaminase polymorphism.

Adenosine modulates cardiovascular functions reducing blood pressure and heart rate. Adenosine deaminase (ADA) by the irreversible deamination of adenosine to inosine contributes to the regulation of adenosine concentration in body fluids. We have studied the interaction between smoking and ADA genetic variability concerning their effects on blood pressure. We have studied 344 subjects admitted to the hospital for cardiovascular diseases. The genotypes of two polymorphic loci within the ADA gene were determined: ADA1 and ADA2. Both loci show two alleles: ADA1*1 and ADA1*2 in ADA1 locus and ADA2*1 and ADA2*2 in ADA2 locus. In the absence of smoking, the proportion of subjects with hypertension tends to be lower in carriers of the ADA1*2 allele. In smoking subjects, the pattern is reversed and the proportion of those with hypertension tends to be higher in carriers of the ADA1*2 allele. A similar pattern is observed for ADA2 locus. Smoking increases the proportion of subjects showing hypertension: such effect is more marked in those carrying the ADA1*2 allele as compared to subjects with ADA1*1/*1 genotype. The same pattern of association is observed for ADA2 locus. The two loci show an additive effect. The odds ratio for hypertension in smokers vs nonsmokers is 1.450 in subjects carrying ADA1*1/*1 and ADA2*1/*1 genotypes, while it is 11.200 in subjects carrying the *2 alleles in both loci. From a practical point, a view of our results suggest that smokers carrying both ADA1*2 and ADA2*2 alleles have a higher risk of hypertension.

Isolated left ventricular non compaction as possible cause of athletic training suspension: a preliminary study on screened athletes.

AIM: The aim of this paper was to determine the prevalence of isolated left ventricular noncomapction (ILVNC) in a sample of 150 athletes send by sports doctors to the Valmontone Hospital's Cardiology Division in a span of about three years, with particular interest in non-compacted segments evaluation. The prevention of cardiovascular complications occurring during sporting activity requires detection of pathologies most often clinically latent but whose first presentation can be sudden cardiac death. In Italy, the pre-participation screening program comprises family history and personal cardiac history, clinical examination and electrocardiography. Subjects with abnormalities are further investigated by stress test, echocardiography and laboratory investigations, and those with significant abnormalities are disqualified from sports training and competition. ILVNC results in multiple trabeculations in the left ventricular myocardium and it is postulated to be caused by intrauterine arrest of compaction of the myocardial fibres and meshwork, an important process in myocardial development. This cardiomyopathy should be considered one of the structural cardiac abnormalities responsible for sudden cardiac death. METHODS: There were 150 athletes seen in the Cardiology Division from 2007 to 2010 for an echocardiographic evaluation in order to clarify the nature of physical examination and/or electrocardiogram abnormalities. Echocardiographic diagnosis of ILVNC was based on criteria published by Jenni et al., and by Stölberger et al. RESULTS: Twenty-four of the 150 tested resulted positive for ILVNC (16.0%). This high prevalence is justified because it was a population originally selected because of electrocardiographic abnormalities. CONCLUSION: We believe that in case of unspecific ECG findings, it would be useful to perform echocardiographic examination in order to highlight structural defects. We also believe that it is very important to contemplate ILVNC between the causes of sudden death in young competitive athletes.

Coronary artery disease: evidence of interaction between PTPN22 and p53 genetic polymorphisms.

OBJECTIVES: We recently reported an association between the PTPN22 genetic polymorphism and coronary artery disease (CAD) in nondiabetic subjects. Since recent studies suggest that p53 may be involved in coronary atherosclerosis, we have investigated a possible interaction between PTPN22 and p53 codon 72 genetic polymorphisms regarding their effects on susceptibility to CAD in nondiabetic subjects. METHODS: The genotypes of p53 codon 72 and PTPN22 were determined by DNA analysis in 128 nondiabetic subjects with CAD, 122 healthy blood donors and 117 nondiabetic subjects with cardiovascular diseases without CAD. RESULTS: In subjects with the *Arg/*Arg genotype of p53 codon 72, no association was observed between CAD and PTPN22. However, this association was very strong in subjects carrying the *Pro allele of p53 codon 72. Subjects carrying both the *T allele of PTPN22 and the *Pro allele of p53 were overrepresented in CAD nondiabetic cases relative to the other two groups (p = 0.001). CONCLUSIONS: Since both p53 and PTPN22 are involved in autoimmune inflammation, an interaction between the two systems appears biologically plausible. In the analysis of multifactorial disorders, the simultaneous analysis of multiple genes functionally related to diseases will provide a more productive approach than studies of single genetic factors performed from a Mendelian perspective.

Atherosclerosis and PTPN22: a study in coronary artery disease.

OBJECTIVES: Recently, it has been shown that PTPN22 genetic polymorphism is associated with phenotypes related to the risk of atherosclerosis. In the present note, we have searched for a possible association of PTPN22 polymorphism with coronary artery disease (CAD). METHODS: One hundred and thirty-four non-diabetic subjects admitted to hospital for CAD and 174 healthy subjects (blood donors) were studied. PTPN22 genotypes were determined by DNA analysis. Statistical analyses were performed by SPSS programs. RESULTS: In CAD patients, the proportion of carriers of the *T allele of PTPN22 is significantly higher compared to healthy controls (OR 2.66; 95% CI 1.07-6.72). CONCLUSIONS: The present observation confirms the association of PTPN22 phenotype with atherosclerosis and suggests a role of immune mechanism in the pathogenesis of CAD.

ACP1 genetic polymorphism and coronary artery disease: an association study.

OBJECTIVES: Assuming an immune component in the pathogenesis of atherosclerosis, we have investigated a possible association between coronary artery disease (CAD) and the acid phosphatase locus 1 (ACP1) genetic polymorphism, which has previously been found to be associated with immune disorders. METHODS: 226 subjects admitted to the hospital for CAD, 358 consecutive newborn infants, 279 adult subjects with type 2 diabetes without CAD and 137 adults without diabetes and without CAD from the Caucasian population of Rome were studied. The ACP1 genotype was determined by DNA analysis. Statistical analyses were performed using the SPSS package. RESULTS: CAD females showed an excess of ACP1 *A/*C and *B/*C genotypes and a deficiency of ACP1 *B/*B genotype compared to controls, while CAD males did not show significant differences. Among diabetic women the proportion of *C allele carriers was much greater in those with CAD than in those without CAD. This difference was much less evident in nondiabetic women. CONCLUSION: ACP1 may be involved in susceptibility to CAD. Since ACP1 has been found to be associated with immunological diseases, our observation reinforces the notion of an immune component in the pathogenesis of atherosclerosis.

Immunodetection of human atherosclerotic plaque with 125I-labeled monoclonal antifibrin antibodies.

To test the affinity of a new F(ab')2 monoclonal antibody (TRF1) against human fragment D dimer of cross-linked fibrin for atherosclerotic plaques free of detectable thrombi, 6 atherosclerotic segments of carotid and femoral artery, and as a control 5 segments of atherosclerosis-free internal mammary artery, were drawn from 11 male patients undergoing bypass surgery. All segments were carefully washed in order to remove possible endoluminal thrombi, and cut to obtain pairs of intimal fragments of similar weight, containing either plaques (n = 16), or fatty streaks (n = 12), or normal endothelium (n = 20). Each fragment underwent a direct binding test to TRF1, or to a non-specific antibody, both labeled with 125I. The activity in each fragment was measured after 3 h of incubation at 37 degrees C, and after washing the fragments every hour for 3 h. TRF1 binding (as percentage of initial activity) was significantly higher (P < 0.001) in atherosclerotic than in normal fragments (26% +/- 11.5%, vs. 9.2% +/- 3.9% in fatty streaks, and 1.9% +/- 0.6% in normal endothelium), and indirect immunofluorescence confirmed TRF1 uptake within the plaque wall. By contrast, the non-specific antibody did not show any significant binding. These preliminary results demonstrate the high specific affinity of TRF1 for atherosclerotic plaques, probably due to the hemorheologic phenomena that activate platelets and provoke the formation of fragment D dimers of cross-linked fibrin on the plaque surface.

Further studies on acid phosphatase in obese subjects.

Low activity genetic variants of acid phosphatase (ACP1) are positively associated with extreme body mass deviations in obese subjects. The same pattern has been found in non-diabetic children, in diabetic pregnant women, and in non-diabetic adult subjects. Low activity variants of ACP1 also show a positive association with family history of obesity, supporting the hypothesis of an enhancing action of these variants on expressivity of obesity.

In vivo study of a technetium labelled anti-EGFr MoAB.

Epithelial Growth Factor receptors (EGFr) are normally present in all the epithelial cells, but their overexpression is closely related to presence of cancer. We have raised EGF-competitive antibody against EGFr and have labelled it with 131I and technetium. The ability of this antibody to bind to A431 cells to be internalized has been tested on A431 cells cultures. Its ability to give scintigraphic images of epithelial tumors has been tested on nu/nu balb c mice xenografted with A431 cells. The labelled antibody is well internalized by cultured cells. Xenografted tumors are clearly imaged both by 131I and 99mTc anti EGFr Mo/Ab. 99mTc labelling is very interesting. The tumor/background ratio was 0.72 +/- 0.2 for 99mTc and 0.40 +/- 0.6 for 131I labelling. Moreover very high uptake of 99mTc MoAb was obtained 2 hours after injection whereas the 131I antibody required 24 hours.

Nitroglycerin-induced changes in myocardial sestamibi uptake to detect tissue viability: radionuclide comparison before and after revascularization.

BACKGROUND: Nitroglycerin (NTG) is known to increase the blood supply to the myocardium, and would thus increase the delivery of a perfusional tracer such as sestamibi (MIBI) to the tissue. The latter, in turn, would take up and concentrate the tracer to a greater extent than in basal conditions only if energy-dependent mechanisms were still available-that is, only if the cells were still viable. METHODS: We evaluated the changes that intravenous administration of NTG induced on the uptake of MIBI by akinetic myocardial areas, using tomographic perfusional imaging in 23 patients with previously ascertained anterior myocardial infarction who were undergoing myocardial revascularization procedures. Changes in uptake were compared with echocardiographic and perfusional changes occurring after operation. RESULTS: The improvement of MIBI uptake after NTG correctly identified 12 of the 16 patients (75%) showing postoperative wall motion improvement; they comprised 12 of the 14 (86%) patients with NTG-induced increase in MIBI uptake who showed improved wall motion after operation. A close correlation (r = 0.88, P < 0.001) was found between the increase in myocardial MIBI uptake induced by NTG infusion and that induced by revascularization. The presence of collaterals to the akinetic area was associated with a significantly (P < 0.01) greater increase in MIBI uptake both during NTG infusion and after operation. CONCLUSIONS: The results of this study suggest that MIBI perfusional myocardial scintigraphy during infusion of NTG is capable of detecting viable but chronically hypoperfused myocardium, predicting postoperative wall motion and perfusional improvement, and reflecting the postoperative pattern of perfusion. The best results were achieved in patients with evidence of collateral circulation supplying the infarcted area.

99Tcm-MIBI uptake in green plants.

Uptake of 99mTcm-sestamibi by biological structures depends on delivery and concentration by electrochemical gradients through the biological membranes and can be simply studied using a green plant model in which photosynthesis tightly modulates water and solute regional flow. Photosynthesis creates electrochemical gradients inside chloroplasts and mitochondria. Moreover, it is the driving force for the movement of water and solutes through induction of pore opening which causes capture of CO2 and loss of water vapour. Thus osmotic pressure increases thereby drawing water from the roots. Hypoestes sanguinolenta was used as an experimental model. This plant displays green zones (with several chloroplasts) and red zones (where they are absent). To detect the uptake differences between these zones we used a new, high-resolution gamma camera. Our results show that (a) 99mTcm-sestamibi is actively transported with water and ions by xylem to leaves where it may diffuse at cellular levels; (b) activation of photosynthesis by light strongly influences the total uptake and the selective compartmentation in green zones; and (c) the green plant's particular physiology tremendously enhances the differences between 99Tcm-sestamibi and 201Tl uptake. We suggest that viable cells, able to create and maintain electrochemical gradients, selectively take up 99Tcm-sestamibi.

Anti D dimer monoclonal antibodies: a possible scintigraphic agent for immunodetection of thrombi.

The aim of this work was to produce a MoAb able to react with clots but not with fibrinogen. Monoclonal antibodies directed towards DD dimers, against specific products of plasmic digestion of cross-linked fibrin, were obtained. One of these antibodies, F 60/43/8, showed a 1.79 x 10(9) l mol-1 binding constant in spite of the presence of fibrinogen at a 4000 times greater concentration than the cross-linked fibrin. In vitro studies with 125I-F(ab')2 of F 60/43/8 showed that 34-80% of the radioactivity can be found in human clots, in the presence of physiologic concentrations of fibrinogen, and that 96-h washing does not remove the labelled F(ab')2 from the clot. 131I-F(ab')2 was injected into rabbits in which a clot had formed in an artery (six rabbits) or in a vein (six rabbits) of the left ear. Scintigraphic images of the clot were always obtained. In conclusion, the results of this work suggest that F 60/43/8 may be used as a specific antibody for the radioimmunodetection of thrombi.

Myocardial scintigraphy with 99mTc-sestamibi in children with Kawasaki disease.

Kawasaki disease (KD) is a systemic vasculitis syndrome of early childhood. It involves particularly the coronary arteries and may cause aneurysms and thrombotic occlusions. Echocardiography is the most useful method of detecting coronary aneurysms. Nevertheless, obstructive lesions are difficult to evaluate and often need invasive coronary angiography. An important feature of this disease is the possibility of finding coronary pathology several years after the onset. This characteristic makes KD an important cause of coronary artery disease (CAD) in young adults. Thus patients with KD and previously diseased coronary arteries should be kept under long-term control. However, coronary angiography is invasive and cannot be performed repeatedly, especially in young patients. As an alternative, thallium 201 scintigraphy has been employed, but its low-energy photons are suboptimal for standard gamma-camera imaging, particularly in children aged less than three years. To verify the usefulness of a noninvasive assessment of myocardial perfusion, the authors used rest and dipyridamole 99mTc-Sestamibi scan in 15 children (ranging from one to six years of age) with Kawasaki's cardiac involvement. Coronary aneurysms have been demonstrated by echocardiography in 12 patients; 8 patients were also submitted to cardiac catheterization. The sensitivity of 99mTc-Sestamibi imaging for detection of overall coronary lesions was 88% and the specificity was 93%. These data suggest that rest/dipyridamole 99mTc-Sestamibi scintigraphy is an accurate and noninvasive method for the detection and follow-up of Kawasaki's cardiac damage even in patients aged one year.

Dipyridamole technetium-99m Sestamibi imaging in the diagnosis of syndrome X.

In a middle-aged woman with anginal chest pain and a normal-appearing angiogram, dypiridamole technetium-99m Sestamibi scintigraphy, a noninvasive method, provided the diagnosis of syndrome X and was used in follow-up to monitor the course of disease.

99mTc SESTAMIBI scintigraphic evaluation of skeletal muscle disease in patients with systemic sclerosis: diagnostic reliability and comparison with cardiac function and perfusion.

The diagnosis of skeletal muscle involvement in patients with systemic sclerosis (SSc) is usually based on clinical, laboratory, electromyographic, and bioptic evidence of muscle disorder, whereas SSc cardiac disease is well established by nuclear medicine techniques (radionuclide ventriculography and myocardial scintigraphy). Previous reports have retrospectively hypothesized a possible relationship between cardiac and muscle involvement in scleroderma patients. In order to improve overall diagnostic accuracy in the qualitative/quantitative assessment of skeletal muscle involvement in these patients and to compare these results with those obtained at the cardiac level, diethylenetriaminepentaacetic acid (DTPA)-99mTc radionuclide ventriculography and 99mTc SESTAMIBI myocardial and muscular scintigraphic examinations were performed in 10 SSc patients and in five healthy subjects. Muscular radioactivity, as assessed at thigh and calf levels by means of a segmental score, was significantly decreased in SSc patients in comparison with healthy subjects (global score value 15.6+/-2.2 vs 22.7+/-1.6, p<0.001), as well as right ventricular ejection fraction (RVEF, 34.3%+/-5.3 vs 53.6%+/-4.2, p<0.001) and myocardial segmental perfusion (global score value, 19.6+/-2 vs 25.9+/-1.1, p<0.01). The results show a high frequency of skeletal muscle involvement in patients with SSc. Moreover, scleroderma patients with muscle disorders, as evidenced by scintigraphy, show a comparable occurrence of cardiac involvement, even in the absence of clinical signs of cardiac dysfunction.

[Role of rectal exploration, suprapubic and transrectal prostatic ultrasonography, and PSA in the diagnosis and follow-up of prostatic carcinoma]. / Il ruolo dell'esplorazione rettale, dell'ecografia prostatica sovrapubica e transrettale e del PSA nella diagnosi e follow-up del carcinoma prostatico.

BACKGROUND AND AIM: The aim of this study was to evaluate the advantages and limits of the various examinations, namely rectal exploration, suprapubic and transrectal scan and PSA, used in the diagnosis and follow-up of prostatic carcinoma. METHODS: The study was carried out in 21 cases of histologically confirmed prostatic carcinoma in patients aged between 57 and 82 years old (mean age: 69.5) referred to the authors' attention between January 1990 and August 1993. RESULTS: With regard to the diagnosis, rectal exploration showed a sensitivity of 80.9%, suprapubic scan 95.2%, transrectal scan and PSA 100%. During the follow-up, patients were divided into operated (9) and non-operated (12) groups. Of the 9 patients undergoing radical prostatectomy, 5 showed residual locoregional disease; of the other 4 who had undergone a complete removal of the gland, one subsequently reported local recidivation. In those patients with residual disease, rectal exploration showed a postoperative sensitivity of 20%, nil sensitivity in the case of local recidivation and 100% specificity in successfully operated patients. Suprapubic scan showed a sensitivity of 60% in patients with residual disease, nil sensitivity in the case of local recidivation and 100% specificity in successfully operated patients. Transrectal scan and PSA revealed 100% sensitivity and specificity in all cases. These patients who were not operated owing to the presence of metastases at the time of diagnosis were divided into those who responded to hormone and chemotherapy (3 total responses, 6 partial responses) and patients who did not respond to this type of treatment (3 non-responders). In the cases of total response, all the tests used obtained 100% specificity. Serum levels of PSA were higher than the threshold value owing to the persistence of metastases. In the cases of partial response to treatment, rectal exploration revealed 50% sensitivity, suprapubic scan 83%, and transrectal scan and PSA 100%. Sensitivity to the four methods used was 100% in all non-responders. CONCLUSIONS: From the results obtained it can be affirmed that the diagnosis of prostate pathology should start with rectal exploration and in the event that this method suggests the probable benignity of the lesion, the diagnostic process should conclude with a suprapubic scan. If rectal examination or suprapubic scan reveal a suspected malignancy, it is essential to perform a transrectal scan or PSA assay which has a high level of sensitivity and specificity for values over 10 ng/ml. During follow-up the only tests which show a high level of sensitivity are transrectal scan and PSA, whereas suprapubic scan and rectal exploration are not reliable in view of the high percentage of false negatives observed. The follow-up of those patients who were not operated and responded totally or partially to treatment must be carried out exclusively using transrectal scan and PSA assay. Suprapubic scan enables the evolution of the neoplasia to be followed over time in those patients who did not respond to treatment.

Muscular uptake of Tc-99m MIBI and TI-201 in Duchenne muscular dystrophy.

Lack of dystrophin, a protein localized to the inner surface of the sarcolemma of the muscle fiber, is the cause of Duchenne type muscular dystrophy. Plasma membrane damage of the muscular fiber occurs, followed by Ca++ influx into the fibers. There is severe mitochondrial damage in dystrophic but still viable fibers. Five children aged 5-7 years were studied with MRI, TI-201, and Tc-99m sestamibi scintigraphy of the thighs. These three methods showed that the sartorius is the least damaged muscle in Duchenne type muscular dystrophy. MRI showed mild damage of adductors and quadriceps; TI-201 scintigraphy showed a marked reduction of radioactivity in the same muscles; Tc-99m sestamibi uptake occurred only in the sartorius muscle; the quadriceps was not imaged and adductors showed a faint image. A decrease of water in muscular fibers as well as fatty fibrous substitution, occurs after death of the fibers, whereas plasma membrane and mitochondrial damage reduced the uptake of tracers when the fiber is still viable. The interesting mismatch between sestamibi and TI-201 can be explained by considering that the cellular mechanism of uptake and retention of Tc-99m sestamibi involves both plasma membrane and mitochondria, whereas the uptake of TI-201 is only affected by plasma membrane damage.

Preliminary evaluation of the usefulness of Tc-99m (V) DMSA in pancreatic neuroendocrine tumors.

The authors describe the possible application of Tc-99m (V) DMSA scintigraphy in pancreatic neuroendocrine tumors. In consideration of the common embryonic origin of these tumors and other neoplastic diseases (medullary thyroid carcinoma, pheocromocytoma, neuroblastoma) that have been well studied with radionuclide imaging, five cases of pancreatic neuroendocrine tumors (two insulinomas, one vipoma, and two unclassified neuroendocrine tumors) were successfully visualized with Tc-99m (V) DMSA scintigraphy, thus giving an overall "imaging confirmation" of the histologic and/or cytologic results in terms of primary and metastatic localization. The authors point out the importance of their results, obtained in a simple and repeatable manner, and suggest a real comparison in this setting between this approach and octreotide scintigraphy.

Reliability of visual and quantitative hepatobiliary scintigraphy in the follow-up of patients who have undergone cholecystectomy and transduodenal sphincteroplasty.

A noninvasive scintigraphic technique to assess the efficacy of a surgical procedure (e.g., cholecystectomy and transduodenal sphincteroplasty) depends on the development of reliable and accurate qualitative or quantitative diagnostic criteria that allow early recognition of the occurrence and site of complications. For this purpose, the authors divided biliary flow into a four-step progression process and analyzed transit times from the peripheral vein to the gallbladder, common bile duct, and duodenum and the transit time from the common bile duct to the duodenum. These quantitative parameters were assessed in nine healthy volunteers and 31 asymptomatic patients who had previous cholecystectomy to validate their reliability. The results indicate that the four-step Tc-99m HIDA progression analysis provides a reliable, noninvasive evaluation of biliary flow, so that it can be applied to patients who have had cholecystectomy.

[Biliary scintigraphy vs. ultrasonography in the etiological diagnosis of acute pancreatitis]. / Biliscintigraphie contre échographie dans le diagnostic étiologique de la pancréatite aiguë.

Etiology of acute pancreatitis has important implications in the treatment of the disease as gallstones pancreatitis requires the correction of the underlying biliary disease. The usefulness of ultrasonography in the detection of stones in emergency has been questioned, and HIDA biliscintigraphy has been reported to be a possible indicator of biliary pancreatitis. This study compares the value of HIDA colesscintigraphy and ultrasonography in the etiologic diagnosis of 35 patients admitted and treated for acute pancreatitis in our Institution. All patients underwent ERCP for the confirmation of the findings. Cholescintigraphy showed no visualisation of the gallbladder, suggesting biliary tract stones, in 25 patients. In all of them ERCP confirmed the presence of gallbladder and/or common bile duct stones, and endoscopic sphincterotomy and later elective cholescystectomy was performed. Ultrasonography failed to demonstrate biliary stones in 11 of those patients. HIDA cholescintigraphy showed a sensitivity and a negative predictive value of 1 vs 0.56 and 0.45 for ultrasonography. From the results of our study it can be concluded that HIDA biliscintigraphy is more reliable than ultrasonography in the discrimination of biliary vs non-biliary acute pancreatitis in emergency.

Is there a role of p53 codon 72 polymorphism in the susceptibility to type 2 diabetes in overweight subjects? A study in patients with cardiovascular diseases.

Two hundred and eighty six subjects with cardiovascular diseases and 147 healthy newborns were studied. P53 codon 72 polymorphism was determined by DNA analysis. The association between BMI and diabetes depends on p53 polymorphism: Odds Ratio shows a high significant association between BMI and diabetes in *Arg/*Arg subjects (p=0.00001). No significant association is observed in *Pro allele carriers (p=0.203).