A comparison of two DNA extraction approaches in the detection of Borrelia burgdorferi sensu lato from live Ixodes ricinus ticks by PCR and reverse line blotting

We tested two approaches to extract Borrelia DNA from live Ixodes ricinus ticks before polymerase chain reaction (PCR) and reverse line blotting (RLB): DNA extraction of one half of the tick after incubation in BSK medium and DNA extraction of the other half of the tick directly, using ammonium hydroxide. Among 2079 ticks, 31.2% (n=649) were found to be Borrelia-infected by PCR-RLB test using at least one of the DNA extraction methods. Five hundred four ticks (24.2%) were found infected after incubation in BSK and 481 (23.1%) after direct DNA extraction from the tick. The difference was not significant. However, these prevalences were significantly lower when only one method was applied (23.1% and 24.2%) compared to the prevalence obtained by the use of both methods (31.2%). In 313 infected ticks discordant results were obtained, i.e., one half of the tick was found to be infected whereas the other half was uninfected. Among these ticks, B. garinii and B. burgdorferi sensu stricto (ss) were significantly more frequently identified in the half tick incubated in BSK. No significant differences were observed for B. burgdorferi ss, B. valaisiana, and for undetermined Borrelia species.     

Live/real time three-dimensional transthoracic echocardiographic assessment of tricuspid valve pathology: incremental value over the two-dimensional technique

Twenty-nine patients with different tricuspid valve (TV) pathologies were studied by both two-dimensional transthoracic (2DTTE) and live/real time three-dimensional transthoracic echocardiography (3DTTE). A major contribution of 3DTTE over 2DTTE was the en face visualization of all three leaflets of the TV in all patients. This allowed accurate assessment of TV orifice area in patients with TV stenosis and carcinoid disease. Loss of TV leaflet tissue, defects in TV leaflets and size of TV systolic non-coaptation could also be delineated and resulted in identifying the mechanism of tricuspid regurgitation (TR) in patients with Ebstein's anomaly and rheumatic heart disease. Prolapse of TV leaflets could also be well visualized and enabled us to develop a schema for systematic assessment of individual segment prolapse which could help in surgical planning. The exact sites of chordae rupture in patients with flail TV as well as right ventricular papillary muscle rupture could be well seen by 3DTTE. 3DTTE also permitted sectioning of various TV masses for more specific diagnosis of their nature. In addition, color Doppler 3DTTE provided an estimate of quantitative evaluation of TR severity, since the exact shape and size of the vena contracta could be accurately assessed. In conclusion, our preliminary experience with 3DTTE has demonstrated substantial incremental value over 2DTTE in the assessment of various TV pathologies.     

Tangerine tomatoes increase total and tetra-cis-lycopene isomer concentrations more than red tomatoes in healthy adult humans

Lycopene, or the foods that contain it, may prevent prostate cancer. Studies suggest that some cis-lycopene isomers are more bioavailable than the trans-lycopene isomer. We hypothesized that tangerine tomatoes, which predominantly contain the tetra-cis isomer, should be a good source of bioavailable lycopene. We fed lunches containing 300 g tangerine or red tomato sauce per day to 21 healthy adults in a double-blind crossover design. We collected blood at baseline and after each treatment and washout period. We measured tetra-cis, other cis, and trans lycopene, as well as other carotenoids, by reversed-phase high-performance liquid chromatography. Both tomato sauces increased lycopene concentrations in blood, but the tangerine tomato sauce caused a greater increase of total and tetra-cis-lycopene. The cis isomer(s) may also have facilitated absorption of the trans-lycopene isomer. Indices of oxidative damage decreased as serum lycopene concentrations increased. Our results suggest that total lycopene concentrations can be increased by substituting tetra-cis-lycopene-rich tangerine tomatoes for common red tomatoes in the diet.     

Severe Paraneoplastic Hyponatremia and Hypoosmolality in a Patient with Small-Cell Lung Carcinoma: Syndrome of Inappropriate Antidiuretic Hormone Secretion Versus Atrial Natriuretic Peptide or Both?

It is well documented in literature that a majority of small-cell lung cancers are associated with paraneoplastic phenomena. We report the case of a 63-year-old man diagnosed with small-cell lung carcinoma, in whom a severe hyponatremia and renal sodium loss with inappropriate antidiuresis were also found during a routine laboratory testing. Syndrome of inappropriate antidiuretic hormone secretion was first suspected in this patient, but another complex pathogenetic mechanism involving atrial natriuretic peptides could be associated, potentiating the deflation of the plasma sodium level. In our patient, the plasma-atrial natriuretic peptide base level, determined with a sensitive radioimmunoassay, was above the normal range (183 pg/mL; normal range, 50 pg/mL, +/- 10 pg/mL), and the antidiuretic hormone plasma level had an oscillatory pattern, varying between 5.5 pg/mL and 7 pg/mL (normal range, 0-4.7 pg/mL). We discuss the pathogenesis and clinical aspects of this association and the therapeutic options for these types of patients.     

The AMPK agonist AICAR inhibits the growth of EGFRvIII-expressing glioblastomas by inhibiting lipogenesis

The EGFR/PI3K/Akt/mTOR signaling pathway is activated in many cancers including glioblastoma, yet mTOR inhibitors have largely failed to show efficacy in the clinic. Rapamycin promotes feedback activation of Akt in some patients, potentially underlying clinical resistance and raising the need for alternative approaches to block mTOR signaling. AMPK is a metabolic checkpoint that integrates growth factor signaling with cellular metabolism, in part by negatively regulating mTOR. We used pharmacological and genetic approaches to determine whether AMPK activation could block glioblastoma growth and cellular metabolism, and we examined the contribution of EGFR signaling in determining response in vitro and in vivo. The AMPK-agonist AICAR, and activated AMPK adenovirus, inhibited mTOR signaling and blocked the growth of glioblastoma cells expressing the activated EGFR mutant, EGFRvIII. Across a spectrum of EGFR-activated cancer cell lines, AICAR was more effective than rapamycin at blocking tumor cell proliferation, despite less efficient inhibition of mTORC1 signaling. Unexpectedly, addition of the metabolic products of cholesterol and fatty acid synthesis rescued the growth inhibitory effect of AICAR, whereas inhibition of these lipogenic enzymes mimicked AMPK activation, thus demonstrating that AMPK blocked tumor cell proliferation primarily through inhibition of cholesterol and fatty acid synthesis. Most importantly, AICAR treatment in mice significantly inhibited the growth and glycolysis (as measured by ¹⁸fluoro-2-deoxyglucose microPET) of glioblastoma xenografts engineered to express EGFRvIII, but not their parental counterparts. These results suggest a mechanism by which AICAR inhibits the proliferation of EGFRvIII expressing glioblastomas and point toward a potential therapeutic strategy for targeting EGFR-activated cancers.     

Noninvasive prediction of tumor responses to gemcitabine using positron emission tomography

Gemcitabine (2′,2′-difluorodeoxycytidine, dFdC) and cytosine arabinoside (cytarabine, ara-C) represent a class of nucleoside analogs used in cancer chemotherapy. Administered as prodrugs, dFdC and ara-C are transported across cell membranes and are converted to cytotoxic derivatives through consecutive phosphorylation steps catalyzed by endogenous nucleoside kinases. Deoxycytidine kinase (DCK) controls the rate-limiting step in the activation cascade of dFdC and ara-C. DCK activity varies significantly among individuals and across different tumor types and is a critical determinant of tumor responses to these prodrugs. Current assays to measure DCK expression and activity require biopsy samples and are prone to sampling errors. Noninvasive methods that can detect DCK activity in tumor lesions throughout the body could circumvent these limitations. Here, we demonstrate an approach to detecting DCK activity in vivo by using positron emission tomography (PET) and ¹⁸F-labeled 1-(2′-deoxy-2′-fluoroarabinofuranosyl) cytosine] ([¹⁸F]FAC), a PET probe recently developed by our group. We show that [¹⁸F]FAC is a DCK substrate with an affinity similar to that of dFdC. In vitro, accumulation of [¹⁸F]FAC in murine and human leukemia cell lines is critically dependent on DCK activity and correlates with dFdC sensitivity. In mice, [¹⁸F]FAC accumulates selectively in DCK-positive vs. DCK-negative tumors, and [¹⁸F]FAC microPET scans can predict responses to dFdC. We suggest that [¹⁸F]FAC PET might be useful for guiding treatment decisions in certain cancers by enabling individualized chemotherapy.     

Ventricular mechanical dyssynchrony and resynchronization therapy in heart failure: a new indication for Fourier analysis of gated blood-pool radionuclide ventriculography

In patients with decreased left ventricular ejection fraction and conduction disease, ventricular mechanical dyssynchrony has been demonstrated. To date, resynchronization by biventricular pacing is increasingly used since it improves ventricular function and exercise capacity in patients with heart failure. To optimize and evaluate the effect of resynchronization therapy and to identify patients who may benefit from biventricular pacing the assessment of left ventricular synchronicity is essential. Therefore, a non-invasive and reproducible technique to obtain information on ventricular synchrony is clinically valuable. In this review, the technical background and the role of phase analysis of gated blood-pool nuclear ventriculography in the assessment of ventricular mechanical synchrony, especially in heart failure patients subjected to biventricular pacing, will be discussed.