Identification and Detection of Phoma tracheiphila, Causal Agent of Citrus Mal Secco Disease, by Real-Time Polymerase Chain Reaction.

Phoma tracheiphila is the causal agent of a tracheomycotic disease of citrus called mal secco causing the dieback of twigs and branches. This pathogen is of quarantine concern; therefore, fast and reliable protocols are required to detect it promptly. A specific primer pair and a dual-labeled fluorogenic probe were used in a real-time polymerase chain reaction (PCR) with the Cepheid Smart Cycler II System (Transportable Device TD configuration) to detect this fungus in citrus samples. Real-time PCR assay was compared to modified conventional PCR assay. The sensitivity of the former was evaluated by testing P. tracheiphila DNA dilutions, and the minimum amount detectable was about 500 fg, whereas the linear quantification range was within 100 ng to 1 pg. Conventional PCR sensitivity was 10 pg. Conventional and real-time PCR successfully detected the fungus in woody samples of naturally infected lemon and artificially inoculated sour orange seedlings. Nevertheless, real-time PCR was about 10- to 20-fold more sensitive than conventional PCR, and preliminary results indicate that the former technique achieves quantitative monitoring of the fungus in tissues. Simple and rapid procedures to obtain suitable DNA samples from fungal cultures and citrus woody samples for PCR assays enable diagnosis to be completed in a short time.

Different patterns of atrial activation in idiopathic atrial fibrillation: simultaneous multisite atrial mapping in patients with paroxysmal and chronic atrial fibrillation.

OBJECTIVES: We aimed to evaluate: 1) the behavior of electrical activity simultaneously in different atrial regions during atrial fibrillation (AF); 2) the difference of atrial activation between paroxysmal and chronic AF; 3) the atrial refractoriness dispersion; and 4) the correlation between the effective refractory periods (ERPs) and the FF intervals. BACKGROUND: Little data exist on the electrophysiologic characteristics of the different atrial regions in patients with AF. A more detailed knowledge of the electrical activity during AF may provide further insights to improve treatment of AF. METHODS: Right and left atria were extensively mapped in 30 patients with idiopathic AF (18 paroxysmal and 12 chronic). In different atrial locations, we analyzed 1) the FF interval duration; and 2) the grade of organization and, in case of organized electrical activity, the direction of atrial activation. Furthermore, in patients with paroxysmal AF, we determined the atrial ERP, evaluated the ERP dispersion and assessed the presence of a correlation between the ERPs and the FF intervals. RESULTS: In patients with chronic AF, we observed a shortening of the FF intervals and a greater prevalence of disorganized activity in all the atrial sites examined. In patients with paroxysmal AF, a significant dispersion of refractoriness was observed. The right lateral wall showed longer FF intervals and more organized atrial activity and, unexpectedly, the shortest mean ERPs. In contrast, the septal area showed shorter FF intervals, greater disorganization and the longest mean ERPs. CONCLUSIONS: Electrical activity during AF showed a significant spatial inhomogeneity, which was more evident in patients with paroxysmal AF. The mean FF intervals did not correlate with the mean ERPs.

Accessory pathway potential recording in a case of permanent junctional reciprocating tachycardia with decremental conduction localized on the atrial site.

Permanent junctional reciprocating tachycardia (PJRT) is an uncommon form of atrioventricular reentrant tachycardia due to the presence of an accessory pathway characterized by slow and decremental retrograde conduction. We report a case of PJRT where we demonstrated the possibility of recording a distinct accessory pathway potential. Decremental retrograde conduction was evident using ventricular extrastimuli and it was also adenosine-sensitive. Delivering ventricular extrastimuli a prolongation of the accessory pathway potential-atrium interval was seen demonstrating that decremental conduction was located at the atrial insertion of the pathway. The accessory pathway was successfully ablated using the potential as the target of radiofrequency delivery. These electrophysiological findings seem to support the hypothesis that a nodal-like structure may be responsible for this arrhythmia.

[Determination, using gas chromatography, of acetonitrile and acrylonitrile in the air]. / Determinazione per via gas-cromatografica di acetonitrile e acrilonitrile nell'aria.

This paper deals with sampling and analysing acetonitrile and acrylonitrile pollutants in the work environments. The sampling procedure in the air is based on -15 degrees C cooled adsorbing solution (water/methanol 65:35 v/v). The quantitative analysis is made by gaschromatograph equipment with Porapak Q 50 divided by 80 mesh column.

Draft genome sequence of Pseudomonas corrugata, a phytopathogenic bacterium with potential industrial applications.

Pseudomonas corrugata was first described as the causal agent of a tomato disease called 'pith necrosis' yet it is considered as a biological resource in various fields such as biocontrol of plant diseases and production of industrially promising microbial biopolymers (mcl-PHA). Here we report the first draft genome sequence of this species.

Draft Genome Sequence of Pseudomonas mediterranea Strain CFBP 5447T, a Producer of Filmable Medium-Chain-Length Polyhydroxyalkanoates.

Pseudomonas mediterranea strain CFBP 5447(T) is a phytopathogenic bacterium isolated from tomato plants affected by pith necrosis disease. Moreover, its ability to produce medium-chain-length polyhydroxyalkanoates (mcl-PHAs) in culture from different carbon sources and valuable microbial products, such as cyclic lipopeptides, has been well documented. Here, we report the first draft genome sequence of this species.

Characterization of isolates of Citrus tristeza virus by sequential analyses of enzyme immunoassays and capillary electrophoresis-single-strand conformation polymorphisms.

Citrus tristeza virus (CTV) is the causal agent of tristeza disease, which is one of the most devastating diseases of citrus crops worldwide. This paper describes a method for the rapid detection and genotyping of naturally spreading CTV isolates. This method uses ELISA or dot-blot immunological tests to detect trees infected with CTV. The reaction wells or membrane spots for which there is a positive reaction are sequentially treated by (i) washing and elution of viral RNA from the trapped samples, (ii) one-step synthesis of cDNA and PCR and (iii) automated fluorescence-based capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) analysis of amplification products. Comparative CE-SSCP results are presented for CTV RNA extracted directly from infected leaves and ELISA plates or from membranes. In the analyses of all of these RNA samples, the p18, p27 and p23 CTV genes were targeted for amplification. Specific profiles of forward and reverse strands were obtained from a group of eight CTV isolates collected in Sicily, each with distinct biological characteristics, which were analyzed using the conventional two-step procedure (immunological detection followed by CE-SSCP molecular characterization after RNA isolation) or in a continuous process of ELISA/CE-SSCP or dot-blot/CE-SSCP starting from infected plant material. The combined method is simple, highly sensitive and reproducible, thus allowing the processing of numerous field samples for a variety of epidemiological needs. The sequential processing of an ELISA or dot-blot/ELISA followed by CE-SSCP is expected to allow the rapid detection of recent CTV infections along with the simultaneous characterization of the genetic diversity and structure of the population of newly invading CTV.

Isolated, anomalous origin of the left anterior descending coronary artery from the right coronary artery with angina pectoris.

The case of a 51-year-old woman with exertional angina pectoris and isolated anomalous origin of the left anterior descending coronary artery from the right coronary artery is reported. This anomalous artery was not narrowed, coursed in front of the pulmonary artery and did not present either anatomical derangement at the take off or intramyocardial course. The mechanism of ischaemia could not be identified.

[Silent ischemic cardiopathy: a study with dynamic electrocardiogram and ergometric test]. / Cardiopatia ischemica silente: studio mediante elettrocardiogramma dinamico e test ergometrico.

Ninety-two patients with effort angina were retrospectively studied to define the prevalence, the daily distribution and the prognostic value of silent ischemic attacks. All patients had positive Holter monitoring and exercise test; coronary angiography, performed in 75/92 patients, showed 1, 2 or 3-vessels disease. Six hundred ninety-three ischemic episodes, 481 (69.5%) silent and 212 (30.5%) symptomatic, were recorded by Holter monitoring, with the highest incidence in the morning; 74/92 patients (80%) showed silent ischemic attacks. Mean duration of the symptomatic and silent ischemic attacks was respectively 9.8 +/- 5.2 and 6.4 +/- 4.2 min (p less than 0.0001); mean ST-segment depression was respectively 2.8 +/- 1.2 and 2.3 +/- 0.8 mm (p less than 0.0001). During exercise testing 86 patients (93%) had both chest pain and ST-segment changes, 2 patients (2%) only angina and 4 patients (5%) only ST-segment depression. Mean heart rate at onset of ischemia was higher during exercise testing compared with Holter monitoring (119 +/- 20 vs 95 +/- 22 b/min; p less than 0.0001). No significant difference was shown between patients with and without silent ischemia about the prevalence of 1, 2 and 3-vessels disease; 1-year cardiovascular mortality in the 2 groups of patients was respectively 6.8% and 5.5% (p:NS). In patients with effort angina, silent ischemia has not a poor prognostic value; Holter monitoring is very useful to the correct assessment of these patients.

Comparison of technetium 99m-tetrofosmin and thallium-201 single photon emission computed tomographic imaging for the assessment of viable myocardium in patients with left ventricular dysfunction.

BACKGROUND: Tetrofosmin is a new technetium 99m-labeled myocardial perfusion agent that has demonstrated favorable imaging characteristics in recent clinical trials. However, it is not certain whether 99mTc-tetrofosmin compared with thallium 201 would underestimate myocardial viability in regions with left ventricular dysfunction. METHODS: To this end 15 patients (mean age 52+/-7 years) with coronary artery disease and left ventricular dysfunction (ejection fraction 35%+/-8%) documented on angiography underwent both quantitative rest-redistribution 201Tl and rest 99mTc-tetrofosmin single photon emission computed tomography imaging. RESULTS; Of 240 total segments on rest-redistribution 201Tl protocol 139 (58%) segments had irreversible 201Tl defects. Of these segments 79 (57%) had only mild to moderate reduction of 201Tl uptake (51% to 85% of normal uptake), whereas the remaining 60 (43%) had severely reduced tracer uptake (< or = 50% of normal uptake). On 99mTc-tetrofosmin protocol 180 (75%) segments had abnormal 99mTc-tetrofosmin uptake; of these segments 120 (67%) had mild to moderate reduction of 99mTc-tetrofosmin uptake, whereas 60 (33%) had severely reduced activity. Among hypokinetic regions concordance between 201Tl and 99mTc-tetrofosmin regarding myocardial viability with a cutoff point of 50% of peak activity was obtained in 28 (90%) of 31 segments (K' = 0.80), leaving only 3 of 31 regions discordant (p = NS). Similarly, among akinetic or dyskinetic regions concordance between the two tracers regarding myocardial viability was achieved in 54 (93%) regions (K' = 0.75), leaving only 4 of the 58 regions discordant (p = NS). CONCLUSIONS: These data show that when the severity of uptake was considered within abnormal segments, a similar amount of 201Tl viable regions were observed by 99mTc-tetrofosmin. Thus these two agents may provide comparable information about myocardial viability when quantitative analysis of defect severity is performed.