Oral Signs and HLA-DQB1∗02 Haplotypes in the Celiac Paediatric Patient: A Preliminary Study.

Celiac disease (CD) diagnosis can be extremely challenging in the case of atypical patterns. In this context, oral signs seem to play a decisive role in arousing suspicion of these forms of the disease. At the same time, the different expressions of the HLA-DQB1∗02 allele apparently seem to facilitate the interpretation of signs and highlighted symptoms. The aim of this work was to verify whether it is possible to identify a correlation between the development of oral signs and different DQ2 haplotypes in celiac pediatric patients. 44 celiac patients with a medium age of 9.9 were studied. Oral examinations were performed in order to identify recurrent aphthous stomatitis (RAS) and dental enamel defects (DED). The diagnosis of DED resulted as being related to allele expression (P value = 0.042) while it was impossible to find a similar correlation with RAS. When both oral signs were considered, there was an increase in correlation with HLA-DQB1∗02 expression (P value = 0.018). The obtained results identified both the fundamental role that dentists can play in early diagnosis of CD, as well as the possible role of HLA haplotype analysis in arousing suspicion of atypical forms of the disease.

Applying proteomic technology to clinical virology.

Developing antiviral drugs, vaccines and diagnostic markers is still the most ambitious challenge in clinical virology. In the past few decades, data from high-throughput technologies have allowed for the rapid development of new antiviral therapeutic strategies, thus making a profound impact on translational research. Most of the current preclinical studies in virology are aimed at evaluating the dynamic composition and localization of the protein platforms involved in various host-virus interactions. Among the different possible approaches, mass spectrometry-based proteomics is increasingly being used to define the protein composition in subcellular compartments, quantify differential protein expression among samples, characterize protein complexes, and analyse protein post-translational modifications. Here, we review the current knowledge of the most useful proteomic approaches in the study of viral persistence and pathogenicity, with a particular focus on recent advances in hepatitis C research.

Cryptic chromosome rearrangements in five patients, with normal and/or abnormal karyotypes, associated with mental retardation, autism and/or epilepsy, detected by BAC genome array-CGH.

This report describes the usefulness of the BAC genome array-CGH platform in the detection of cryptic rearrangements. We examined ten patients with normal and/or abnormal karyotypes and dysmorphic features, associated with mental retardation, autism and/or epilepsy. This approach led us to discover further cryptic chromosomal rearrangements, not previously detected by conventional cytogenetic procedures, and allowed us to better delineate genotype/phenotype correlation. Our experience shows the validity of the BAC platform as a reliable method for genome-wide screening of chromosomal aberrations in patient with idiopathic mental retardation and/or in association with autism and epilepsy.

A rapid sample method for HLA haplotype typization. A preliminary study on celiac patients.

AIM: The aim of the present work was to determine the human leukocyte (HLA) haplotype in 64 Sardinian patients affected with celiac disease, using a rapid and easy to apply sampling method that permits samples from blood drawing to be stored more easily. Numerous studies have demonstrated how the HLA system plays a very important role in immune system regulation, determining a link between this gene and a high number of pathologies including celiac disease. In fact a genetic susceptibility exists in celiac sprue, linked to HLA-DQB1*0201 and -DQB1*0302 genes which represent sierologic groups -DQ2 and -DQ8 whose early identification could be fundamental in obtaining a diagnosis of celiac disease. METHODS: To realize this study a collecting method of samples was developed through the brushing of oral mucosa, which is extremely less traumatic than the classic sampling method using blood drawing, and which also allows a long conservation period before sample analysis. Samples were later analyzed with Van Embden's DNA extraction method to extract the patient's DNA, on which we executed the Polymerase Chain Reaction (PCR). To obtain the HLA haplotype from each patient we used 8 specific primers that amplified the HLA-DQB1 allele in low-resolution. RESULTS: Out of the 64 patients we found 26 HLA-DQB1*02 homozygotes, 28 HLA-DQB1*02 heterozygotes and 10 negative samples for the HLA-DQB1*02 allele, thus confirming what had emerged from previous blood draws. CONCLUSION: These results show how the method we developed using oral brushing is a sure method to obtain samples for determining the HLA haplotype in extra-hospital areas. This could allow the use of this method to obtain early diagnosis for chronic pathologies linked to the HLA groups and for recognizing this genotype in extensive population studies.

Bronchoalveolar glutathione and nitrite/nitrate in idiopathic pulmonary fibrosis and sarcoidosis.

BACKGROUND AND AIM OF THE WORK: Bronchoalveolar lavage (BAL) is useful in diagnosis and management of interstitial lung diseases. Glutathione (GSH) represents an important defence molecule against reactive oxygen species produced during inflammation, which underlies both idiopathic pulmonary fibrosis (IPF) and sarcoidosis. Nitric oxide has been suggested as a marker of airway inflammation. The aim of this study was to examine the content of GSH and NO stable metabolites, nitrite/nitrate, in the BAL of a group of patients with IPF and sarcoidosis. METHODS: BAL was performed in 13 patients with newly diagnosed IPF, 13 patients with sarcoidosis and 4 controls. Total GSH and nitrite/nitrate were measured. BAL was repeated, in 5 patients with IPF, after 6 and 12 months and GSH was measured again. RESULTS: IPF patients had significantly lower levels of total GSH compared to sarcoidosis patients or controls (129 +/- 22 vs. 324 +/- 40 and 383 +/- 23 mM; p < 0.001) in the epithelial lining fluid (ELF). In patients with IPF (n = 5), total GSH levels in the ELF increased after 6 and 12 months (T0 = 98 +/- 19; T6 = 219 +/- 22; T12 = 301 +/- 34 microM; p < 0.05) following therapy with immunosuppressive drugs such as corticosteroids, methotrexate, cyclosporin A. Levels of nitrite/nitrate were found significantly elevated in IPF (n = 8) and sarcoidosis (n = 10) patients compared to controls (230 +/- 37 and 216 +/- 36 vs. 86 +/- 11 microM; p < 0.05). CONCLUSION: These observations support the role of inflammation and antioxidant defences in interstitial lung diseases and may provide insights into the pathogenesis of oxidant-induced interstitial disease and their therapy.

[Microcystic lateral parodontal cyst. Report of a case]. / Cisti parodontale laterale microcistica. Descrizione di un caso clinico.

The lateral periodontal cyst (LPC) is an odontogenetic lesion of the maxillary bone representing about 0.8% of maxillary cystic lesions. It starts in the lateral periodontal area of a vital erupted tooth. Clinical symptoms are generally absent. Its diagnosis usually can be made on the basis of an occasional radiographic examination which shows an osteolytic monoloculate lesion with defined outlines. The botryoid cyst is a variation of LPC and is radiographically different and more aggressive than the common LPC. A clinical case of LPC which allows to make histopathological and clinical considerations on these cystic lesions is described.

Central nervous system involvement in systemic lupus erythematosus: cerebral imaging and serological profile in patients with and without overt neuropsychiatric manifestations.

The aim of this study was to evaluate morphological and functional abnormalities by cerebral imaging in a series of systemic lupus erythematosus (SLE) patients with and without overt central nervous system (CNS) manifestations, and to detect possible relationships with clinical parameters and a large panel of autoantibodies, including those reactive against neurotypic and gliotypic antigens. 68 patients with SLE were investigated in a cross-sectional study which included clinical evaluation of symptoms, cerebral magnetic resonance imaging (MRI) and brain single photon emission tomography (SPECT) analysis, electroencephalography (EEG), and serological tests for antibodies directed against nuclear, cytoplasmic neuronal and glial cell-related antigens. The results of this study showed: (1) a significant positive association of (a) anti-glial fibrillary acidic protein (GFAP) serum antibodies with neuropsychiatric (NP) manifestations and (b) anti-serin proteinase 3 (anti-PR3/c-ANCA) serum antibodies with pathological cerebral SPECT; (2) the presence of significantly higher values of (a) SLICC organ damage index in patients with abnormal MRI and (b) SLAM activity index in patients with abnormal SPECT; and (3) the association of (a) abnormal MRI with nonactive NP manifestations and (b) combined abnormality of brain SPECT and MRI with the occurrence of overall overt NP manifestations and with those of the organic/major type. Neuropsychiatric manifestations, namely those of the organic/major type, appeared to be significantly associated to the presence of a serum antibody against GFAP, a gliotypic antigen. There was also evidence of an association between SPECT abnormality and the presence of anti-PR3 (c-ANCA). Furthermore, brain imaging by MRI and SPECT applied to SLE patients appears to express CNS involvement significantly related to specific categories of NP manifestations. The abnormalities detected by the two tests seem to be preferentially associated with different activity phases of the NP disorder or of the lupus disease.

Lack of melatonin effect on hydrogen peroxide induced bronchoconstriction in isolated and perfused rat lung.

The effect of melatonin on hydrogen peroxide- induced broncho-and vasoconstriction was examined in vivo in the model of the isolated, perfused and ventilated lung. The administration of hydrogen peroxide (500 microM) to the perfusate caused a marked decrease in lung compliance, conductance and flow rate. The administration of melatonin (500 microM) to the perfusate 20 min before and during the hydroperoxide exposure did not cause any change in lung function. Exposure of lung microsomes to hydrogen peroxide (1-100 microM) did not induce any significant increase in malonaldehyde (MDA), an index of lipid peroxidation, and it was not affected by treatment with melatonin (500 microM). On the other hand, brain microsomes exposed to hydrogen peroxide (1-100 microM) give rise to increased levels of MDA, which were decreased by pre-treatment with melatonin (500 microM). The results suggest that melatonin may exert an antioxidant effect in conditions were lipid peroxidation is occurring. Its use may not be relevant in conditions where the mechanisms of the reactive oxygen species damage appears to be lipid peroxidation independent, such as the case of hydrogen peroxide induced broncho- and vasoconstriction.

Correlation of 99mTc-sestamibi SPECT myocardial perfusion imaging with absolute coronary flow reserve by a new noninvasive Doppler method in patients with stenoses of the left anterior descending coronary artery. A preliminary report.

BACKGROUND: Myocardial perfusion assessed by a pharmacological scintigraphic test is a time-honored method to assess the functional significance of a coronary stenosis. There is no study that has compared coronary flow reserve assessed by adenosine/base contrast-enhanced transthoracic echo Doppler in harmonic mode with myocardial perfusion by adenosine/base 99mTc-sestamibi SPECT. The aim of this study was to compare the absolute coronary flow reserve measured in the left anterior descending coronary artery using a new noninvasive method (transthoracic harmonic Doppler) with adenosine/base 99mTc-sestamibi SPECT. METHODS: Twenty-two patients scheduled for coronary angiography underwent evaluation of basal and hyperemic flow in the left anterior descending coronary artery using transthoracic harmonic Doppler. Simultaneously (during the same adenosine infusion) the patients were injected with 99mTc-sestamibi (10 mCi). Baseline scintigraphic study was obtained 4 hours later (99mTc-sestamibi, 30 mCi). RESULTS: Angiography showed in the left anterior descending coronary artery no or not significant stenosis (< 40% lumen narrowing) in 14 patients (Group I) and a severe stenosis (> 75%) in the remaining 8 patients (Group II). Coronary flow reserve dichotomized as normal (> or = 2.0) or abnormal (< 2.0) gave concordant results with SPECT. Overall, in fact, 18 studies (81%) were in agreement (k = 0.48, p = 0.02). In addition, coronary flow reserve proved to be a useful complementary tool to SPECT in patients with severe multivessel disease (and global hypoperfusion; 2 cases) and left bundle branch block (1 case). CONCLUSIONS: Coronary flow reserve assessed by transthoracic harmonic Doppler gives concordant results with SPECT perfusion scintigraphy. The diagnostic potential of SPECT may be increased by the noninvasive assessment of coronary flow reserve in the left anterior descending coronary artery.

Validation of a new noninvasive method (contrast-enhanced transthoracic second harmonic echo Doppler) for the evaluation of coronary flow reserve: comparison with intracoronary Doppler flow wire.

OBJECTIVES: We tested the hypothesis that coronary flow reserve (CFR) in the left anterior descending coronary artery (LAD) as assessed by a new noninvasive method (contrast-enhanced transthoracic second harmonic echo Doppler) is in agreement with CFR measurements assessed by intracoronary Doppler flow wire. BACKGROUND: Contrast-enhanced transthoracic second harmonic echo Doppler is a novel noninvasive method to detect blood flow velocity and reserve in the LAD. However, it has not yet been validated versus a gold-standard method. METHODS: Twenty-five patients undergoing CFR assessment in the LAD by Doppler flow wire were also evaluated by contrast-enhanced transthoracic Doppler to record blood flow in the distal LAD at rest and during hyperemia obtained by adenosine i.v. infusion. In five patients CFR was evaluated twice (before and after angioplasty). RESULTS: As a result of the combined use of i.v. contrast and second harmonic Doppler technology, feasibility in assessing coronary flow reserve equaled 100%. The agreement between the two methods was high. In fact, in all but five patients the maximum difference between the two CFR measurements was 0.38. Overall, the prediction (95%) interval of individual differences was -0.69 to +0.72. Reproducibility of CFR measurements was also high. The limits of the agreement (95%) between the two measurements were -0.32 to +0.32. CONCLUSIONS: Coronary flow reserve in the LAD as assessed by contrast-enhanced transthoracic echo Doppler along with harmonic mode concurs very closely with Doppler flow wire CFR measurements. This new noninvasive method allows feasible, reliable and reproducible assessment of CFR in the LAD.

Contrast-enhanced transthoracic second harmonic echo Doppler with adenosine: a noninvasive, rapid and effective method for coronary flow reserve assessment.

OBJECTIVES: The purpose of this study is to evaluate the feasibility in detecting blood flow in the left anterior descending coronary artery (LAD) using transthoracic color Doppler (CD) imaging (in both second harmonic and fundamental mode) along with contrast enhancement and to verify if this new noninvasive method along with adenosine is safe, rapid and effective in assessing coronary flow reserve (CFR). BACKGROUND: Feasibility of contrast-enhanced transthoracic Doppler recording (in both second harmonic and fundamental mode) of blood flow velocity in the LAD has not been assessed. Adenosine has a greater vasodilator potency and more favorable kinetics than dipyridamole and thus it can be more suitable for assessing CFR in conjunction with this method. METHODS: Sixty-one patients with angiographically patent LAD underwent CD (both in fundamental and harmonic mode) as well as color-guided pulsed wave (PW) Doppler recording of blood flow velocity in the distal LAD before and after intravenous contrast injection. A second group of patients (n = 77), undergoing coronary angiography, was submitted to transthoracic contrast-enhanced PW Doppler recording of blood flow velocity in the LAD using harmonic CD as a guide, at rest and during adenosine-induced hyperemia. RESULTS: Harmonic CD along with echo contrast consistently improved blood flow detection in the LAD; the success rate in detecting flow of optimal quality was 88% with this approach, whereas it was 11% and 16% with CD in fundamental mode, respectively, before and after contrast. Pulsed wave Doppler results paralleled those of harmonic CD (p < 0.001 contrast harmonic vs. fundamental). In the second group of patients coronary angiography revealed 0% to <40% stenosis in 24 patients (group I), > or =40% to < or =75% in 17 patients (group II) and >75% stenosis in 34 patients (group III). There was a significant difference in CFR among the three groups of patients; CFR for peak diastolic velocity was (mean +/- SD): 2.88+/-0.7 (group I), 2.09+/-0.5 (group II) and 1.51+/-0.5 cm/s (group II) (p < 0.05 group I vs. both group II and group III; p < 0.05 group II vs. group III). The whole examination took less than 10 min. CONCLUSIONS: Contrast-enhanced second harmonic Doppler recording of blood velocity in the LAD is highly feasible and in combination with adenosine it is a rapid, safe and effective method for assessing CFR ratio.

New noninvasive method for coronary flow reserve assessment: contrast-enhanced transthoracic second harmonic echo Doppler.

BACKGROUND: We tested the hypothesis that blood flow velocity could be recorded in the left anterior descending coronary artery (LAD) during transthoracic echocardiography by use of second harmonic echo Doppler modality along with contrast enhancement (intravenous Levovist) at rest and after pharmacologically induced maximal vasodilation to assess coronary flow reserve (CFR) with a totally noninvasive approach. METHODS AND RESULTS: Fifty-six consecutive patients undergoing coronary angiography underwent transthoracic contrast-enhanced pulsed-wave Doppler recording of blood flow velocity in the LAD by use of harmonic color Doppler as a guide at rest and after maximal vasodilation by dipyridamole infusion. Contrast enhancement with the harmonic mode greatly improved the success rate of recording adequate pulsed-wave Doppler signal in the LAD. CFR was (mean+/-SD) 1.54+/-0.7 in patients with (group 1) and 2. 79+/-0.9 in patients without (group 2) significant LAD stenosis (lumen narrowing >70%) (P<0.001); sensitivity and specificity in detecting significant LAD stenosis were 86% and 90%, respectively. There was close agreement between CFRs determined by this new method and intracoronary Doppler flow wire. CONCLUSIONS: Contrast-enhanced transthoracic echo Doppler with the harmonic mode is a feasible and promising technique for assessing CFR in a totally noninvasive way.

Involvement of capsaicin-sensitive nerves in paraquat-induced mortality.

Paraquat (PQ), a broad spectrum herbicide, produces severe lung inflammation and necrosis resulting in pulmonary fibrosis and respiratory failure. Tachykinins are peptides released by sensory C fibers and have the ability of influencing respiratory functions and cellular proliferation. To examine whether the damage caused by PQ involves tachykinins, rats were depleted in their content of tachykinins by systemic treatment with capsaicin prior to PQ exposure. The animal subjected to this treatment showed a 3-fold higher viability compared to those treated with PQ alone (75 vs 27%). Depletion of reduced glutathione (GSH) is associated with oxidative stress produced by reactive oxygen intermediates during PQ metabolism. This is considered to be critical in the pathogenesis of lung damage by PQ. PQ treatment induced a significant depletion of GSH during the first days and a similar effect was also observed in the group of capsaicin-pretreated rats. Four weeks after PQ treatment the levels of GSH were similar to controls in rat pretreated or not with capsaicin plus PQ. This may indicate that the reduced levels of GSH may be associated to the toxicity observed in the acute phase, but not of importance in the final PQ-induced mortality. Neutral endopeptidase (NEP) is an enzyme considered to be critical in controlling the levels of tachykinins. Exposure of crude membrane preparations of rat lung to PQ resulted in a dose-dependent inhibition of NEP activity. Since NEP inactivation may occur in lung following a PQ exposure in vivo, the results indicate that during PQ intoxication a more sustained activity of tachykinins may be present, producing effects such as cell proliferation, fluid extravasation and bronchoconstriction. In conclusion, this finding supports the hypothesis that neuropeptides released from capsaicin-sensitive nerves could be involved in the modulation of PQ-induced lung damage.

Early detection of the anthracycline-induced cardiotoxicity. A non-invasive haemodynamic study.

Anthracyclines are the most frequent cause of iatrogenic congestive heart failure ranging from acute reversible minor, irreversible reduction in the left ventricular ejection fraction and death despite preventive measures. Sensitive methods are needed to detect earliest preclinical cardiotoxicity along with the development of new protective agents. Thirty breast cancer patients were randomly treated with q 21 120 mg/m2 Epirubicin (EPI) x 3, alone (10 patients), or + ICRF-187 (1000 mg/m2) (10 patients) or + C0Q10 (50 mg/day) (10 patients) and monitored by Thoracic Electrical Bioimpedance (TEB) cardiography before (T0) and at the end of chemotherapy (T1), then at 1, 3, 6 months of follow up (F1, F2, F3). a) The group treated with EPI alone showed, between F1-F2, a significant (p < 0.05) decrease in Stroke Index (S1). Acceleration Index (ACI) and a significant (p < 0.05) increase in Systemic Vascular Resistance Index (SVRI), while between F2 and F3 it showed a significant (p < 0.05) recovery in S1 and ACI. b) The group treated with EPI + ICRF-187 showed, between F1 and F2 a significant decrease in S1 and ACI (p < 0.05, p < 0.01 respectively) and a significant (p < 0.05) increase in SVRI: between F2-F3 ACI had a significant (p < 0.05) recovery: c) The group treated with EPI +C0Q10 showed no modification in Sl, ACI, and SVRI during the study. The ejection Fraction (EF) remained unchanged during the study in all the groups. C0Q10 seems to prevent early decreases in cardiac performance and contractiling, thus avoiding an SVRI increase, while ICRF-187 did not. Since ICRF-187 acts by binding iron, we deem that the earliest cardiac involvement, may occur before iron overload; therefore the role of ICRF-187 and C0Q10 in acute or chronic heart toxicity was correlated with high-dose anthracycline and needs to be further investigated.

[Hemodynamic assessment at rest and during dynamic physical exercise in young subjects with and without hypertensive parents]. / Comportamento dei parametri emocardiodinamici a riposo e durante escercizio fisico dinamico in giovani soggetti con e senza familiarità ipertensiva.

The aim of this research was to identify any early cardiovascular changes that may be predictive of future hypertension in young subjects with family history of hypertension. The study was conducted on 25 offspring of hypertensive parents, mean age 17 years (22 with hypertension only in 1 parent and 3 with both hypertensive parents) and 20 offspring of both normotensive parents, matched by age. Subjects were divided into children (7-13 years) and young adults (19 years on). All subjects underwent three office blood pressure measurements with a mercury sphygmomanometer. On the third control, BoMed thoracic electrical bioimpedance at rest and during upright bicycle exercise was performed. Physical characteristics were similar in subjects matched by age in the two groups. Systolic blood pressure was similar in offspring of normotensives and hypertensives, both at rest and during exercise; diastolic blood pressure was greater in offspring of hypertensive parents at rest (73.1 +/- 10.5 vs 63.5 +/- 7.1 mmHg, p < 0.05), during the first minutes of exercise and during the recovery phase (p < 0.05). Moreover, at the third blood pressure measurement at rest, diastolic blood pressure decreased, with respect to the first measurement, only in children and young adult offspring of normotensive parents, while systolic blood pressure decreased in the two groups of child subjects. No differences in heart rate were observed, both at rest and during physical exercise, between offspring of normotensives and hypertensives. Left ventricular end-diastolic volume, stroke volume, ejection fraction, cardiac output and systemic vascular resistance at rest and their response to decubitus changes and exercise were normal and similar in offspring of normotensive and hypertensive parents both in children and young adults. In conclusion, a different behavior of diastolic blood pressure was found in offspring of hypertensive parents compared to that of normotensive parents, both in children and, to a higher degree, in young adults. This may be an expression of early vascular change in subjects with a genetic predisposition to hypertension.

Abnormalities of magnetic resonance imaging of the central nervous system in patients with systemic lupus erythematosus correlate with disease severity.

Forty randomly selected patients with systemic lupus erythematosus (SLE) were studied by clinical and serologic parameters and magnetic resonance imaging (MRI). Abnormal MRI was found in 15/40 patients (37.5%): all 15 cases showed multiple widespread small-sized areas of increased signal in T2 in the white matter; in one of these patients MRI also displayed a large area with a reduced signal in T1 and an increased signal in T2 involving both the white and the gray matter. Among the 15 patients with abnormal MRI, only 7 had neuropsychiatric symptoms. The presence of MRI changes was highest in patients with organic type symptoms and was associated to the highest disease severity scores. A long-term follow up of asymptomatic patients would be useful to establish whether the application of MRI is appropriate for the assessment of CNS involvement in SLE.

Multiple cerebral hydatid disease: case report with magnetic resonance imaging study.

A case of multiple hydatid cysts of the brain is reported. Computed tomographic and magnetic resonance imaging scanning revealed a thick and calcified pericystium. These findings allowed appropriate surgical planning, with a marked recovery of the neurological deficits and absence of relapse in the follow-up period.

[Diagnosis of hepatic hemangioma with simplified double 99mTc labelling]. / La diagnosi di emangioma epatico mediante doppia marcatura semplificata con 99mTc.

The authors report on their experience with liver hemangioma (LH) diagnosis by means of a simplified method--that is, the simultaneous, in vivo, double labelling of liver reticuloendothelial system (RES) and of red blood cells (RBC) by 99mTc. Twenty-eight patients with US diagnosis of suspected LH and 15 controls were examined after sequential iv injection of SnCl2, of 99mTc-mucolloid albumin and, after liver scintigraphy, of 99mTc-pertechnetate to conclude in vivo RBC labelling. All patients underwent CT and, if necessary, CT-guided biopsy. Focal colloid defects filled after RBC labelling were shown in 20/22 patients with unquestionable LH. No colloid defects were shown in 6/28 cases (expansive process). 15/15 controls showed unchanged non-filling defects after double labelling. Finally, the authors point out that, in the diagnosis of LH, sequential double labelling of liver RES and RBC appears to be a quicker scintigraphic technique than conventional ones. Moreover, this technique has the same high specificity and sensitivity as more time-consuming ones.