Surveillance of active human cytomegalovirus infection in hematopoietic stem cell transplantation (HLA sibling identical donor): search for optimal cutoff value by real-time PCR

<p>Abstract</p> <p>Background</p> <p>Human cytomegalovirus (CMV) infection still causes significant morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). Therefore, it is extremely important to diagnosis and monitor active CMV infection in HSCT patients, defining the CMV DNA levels of virus replication that warrant intervention with antiviral agents in order to accurately prevent CMV disease and further related complications.</p> <p>Methods</p> <p>During the first 150 days after allogeneic HSTC, thirty patients were monitored weekly for active CMV infection by <it>pp65 </it>antigenemia, nested-PCR and real-time PCR assays. Receiver operating characteristic (ROC) plot analysis was performed to determine a threshold value of the CMV DNA load by real-time PCR.</p> <p>Results</p> <p>Using ROC curves, the optimal cutoff value by real-time PCR was 418.4 copies/10⁴PBL (sensitivity, 71.4%; specificity, 89.7%). Twenty seven (90%) of the 30 analyzed patients had active CMV infection and two (6.7%) developed CMV disease. Eleven (40.7%) of these 27 patients had acute GVHD, 18 (66.7%) had opportunistic infection, 5 (18.5%) had chronic rejection and 11 (40.7%) died - one died of CMV disease associated with GVHD and bacterial infection.</p> <p>Conclusions</p> <p>The low incidence of CMV disease in HSCT recipients in our study attests to the efficacy of CMV surveillance based on clinical routine assay. The quantification of CMV DNA load using real-time PCR appears to be applicable to the clinical practice and an optimal cutoff value for guiding timely preemptive therapy should be clinically validated in future studies.</p

Withdrawal of maintenance therapy for cytomegalovirus retinitis in AIDS patients exhibiting immunological response to HAART

BACKGROUND: Before the introduction of highly active antiretroviral therapy (HAART), CMV retinitis was a common complication in patients with advanced HIV disease and the therapy was well established; it consisted of an induction phase to control the infection with ganciclovir, followed by a lifelong maintenance phase to avoid or delay relapses. METHODS: To determine the safety of CMV maintenance therapy withdrawal in patients with immune recovery after HAART, 35 patients with treated CMV retinitis, on maintenance therapy, with CD4+ cell count greater than 100 cells/mm³ for at least three months, but almost all patients presented these values for more than six months and viral load < 30000 copies/mL, were prospectively evaluated for the recurrence of CMV disease. Maintenance therapy was withdrawal at inclusion, and patients were monitored for at least 48 weeks by clinical and ophthalmologic evaluations, and by determination of CMV viremia markers (antigenemia-pp65), CD4+/CD8+ counts and plasma HIV RNA levels. Lymphoproliferative assays were performed on 26/35 patients. RESULTS: From 35 patients included, only one had confirmed reactivation of CMV retinitis, at day 120 of follow-up. No patient returned positive antigenemia tests. No correlation between lymphoproliferative assays and CD4+ counts was observed. CONCLUSION: CMV retinitis maintenance therapy discontinuation is safe for those patients with quantitative immune recovery after HAART

Comparative performance of medium voltage overhead distribution lines designs submitted to induced voltages

The aim of this paper is to present the results of the performance study of medium voltage overhead distribution lines against lightning discharges, in the way to define methodologies to reduce the system failures. The results are obtained within the partnership among the High Voltage Laboratory of the Federal University of Itajubá, AES Sul Utility Company and the University of Bologna. The resultant performance is presented in terms of expected faults for 100 km of line for a density of discharges to the ground (GDF) of 1 discharge/km2/year. Commentaries on the relative performance and comparisons of different construction configurations of overhead lines are presented. ©2007 IEEE

Quality control of poly(methyl methacrylate) to medical purpose by multiple headspace extraction

The quality of polymers to be used for medical purposes is evaluated by the concentration of residual compounds in the polymeric matrix, especially by the amount of residual monomer. Residual components of the polymerization of monomer can cause allergies and biological complications (stomatitis, dermatitis, cheilitis, and irritability), also collateral effects for the patient which are evident from the first five years after implant placement and could remain for thirty years more. In dentures, for example, these components are dissolved by the saliva and are fixed to adjacent tissue causing allergic reactions, including burning. Multiple headspace extraction gas chromatography (MHS-GC) has been found to be an analytical technique particularly suitable for quantification of residual monomer in process samples with complex matrix, including solids. The major advantage of MHS-GC is that there is no need to pre-treat the sample prior to analysis. In this work, the methodology used to determine the residual monomer of Methyl Methacrylate (MMA) in a polymer of Poly(Methyl Methacrylate) (PMMA) is presented. The PMMA was produced in a controlled pilot plant scale laboratory, with rigorous experimental conditions to be used for medical purposes (artificial bone). The method includes the formulation of a calibration curve which was obtained by injecting different masses (0 - 30 mg) of a standard MMA solution in the Headspace sample vials and treated at ten extraction step in the HS-GC system. The results showed that through the present method it is possible to recover 98% of MMA from a solid matrix of PMMA

miR-501 overexpression in renal carcinomas

Background: MicroRNAs (miR) are small, noncoding RNAs (20–23 nucleotides) that regulate gene expression. In particular, miRs are involved in many biological processes, including cellular differentiation, proliferation and death. In addition to their physiological functions, miRs are found to be aberrantly expressed in certain carcinomas and to play oncogenic or tumor suppressive roles in neoplastic cells (1). One in 75 people will develop renal cell carcinoma (RCC) during their lifetime being the seventh leading cause of cancer in men and eighth in women in the world. RCC is grouped in different types where the clear cell carcinoma (ccRCC) is the most frequent (2). Since little is known on the role of miRs in renal tumorigenesis, we screened, for some miRs expression, the most common renal carcinomas as clear cell (ccRCC), papillary (pRCC) and chromophobe (chRCC). We investigated miR-501, 196b and 202 that we found overexpressed in Polycystic kidney disease, a hyperproliferative renal disease. In addition, miR-501 was found predominantly expressed in mouse colon, lung and kidney. Methods: We selected and analyzed 41 tumor samples: 24 post-nephrectomy fresh frozen tissues (including 13 ccRCC, 7 pRCC, 1 chRCC, 1 lyposarcoma and 2 oncocytomas) and 17 paraffin-embedded samples (12 ccRCC, 4 pRCC, 1chRCC). Total RNA was extracted with RNeasy Plus Kit (fresh frozen tissues) or the RecoverAll® Total Nucleic Acid Isolation kit (paraffin-embedded tissues). Quantitative Real-time PCR for mature miRs was performed with TaqMan method. Levels of miR expression were calculated as DeltaDeltaCt (DDCt) method by using U6 snRNA as reference, and they were related to those of normal tissue as fold change (2-DDCt). Results: No variations were observed in the expression of miR 196b and 202 in RCC compared to normal tissues. Levels of miR-501 were instead expressed in a different manner in post-nephrectomy fresh-frozen tumors as well as in paraffin-embedded tissues compared to non neoplastic tissues. In four patients miR-501 expression was evaluated in both frozen and paraffin-embedded tissues to look for possible variability due to the different storage. Expression levels found in paraffin-embedded tissues were overlapping with those obtained in frozen tissues. Based on miR-501 expression, renal carcinomas were divided in four groups: 1. lover expression, 5 times. Overall, miR-501 expression was downregulated (group 1) in 12 patients (3 ccRCC, 7 pRCC, 1 chRCC and 1 oncocytoma); was normal in 10 patients (7 ccRCC, 2 pRCC and 1 lyposarcoma); weakly over-expressed in 7 samples (4 ccRCC, 1 pRCC, 1 oncocytoma and 1 chRCC); strongly over-expressed in 9 ccRCC samples. No correlation between miR-501 expression and tumor grading was observed. Conclusions: Our results demonstrate that in renal carcinomas the miR-501 was mainly over-expressed in ccRCC, while it was mainly unchanged or down-regulated in chRCC and pRCC. The overexpression of miR-501 in ccRCC, therefore, may contribute to features of this cancer playing a role in therapeutic response, metastasis development and survival. References: 1. Di Leva G, Croce CM: Roles of small RNAs in tumor formation. Trends Mol Med. 16(6):257-67, 2010. Review. 2. Brannon AR, Rathmell WK: Renal cell carcinoma: where will the state-of-the-art lead us? Curr Oncol Rep. 12(3):193-201, 2010. Review

Performance of Medium Voltage Overhead Distribution Lines Against Lightning Discharges

The lightning discharges are one of the main causes of interruptions of medium voltage overhead distribution lines, being the reason of great concern for the utility companies. Its destructive effects frequently extends to equipments and connected installations, with the possibility to cause personal injuries and material damages, beyond economic losses, due to out of income and the possibility of indemnities, penalties and fines. With the beginning of the deregulation of the electric energy supply, some actions were been taken by the utilities for the prevention and minimization of the damages associated to the lightning discharges. However, as the lightning discharges are random events, consequently, difficult to predict, the majority of these actions does not follow a study or a detailed analysis of the problem. By this way, in the majority of the cases the actions were taken based on the knowledge of the engineer in charge, or based in rules defined without any effective evidence, by means of studies or by laboratory tests. As a result, many of them besides of presenting high cost are not effective. Among others, the distribution network reliability depends directly on its exposition to the lightning discharges. To determine the exposition level of the line, the designer should know the number of discharges to the ground per unit of area per unit of time. The aim of this paper is to present the results of the performance study of medium voltage overhead distribution lines against lightning discharges, in the way to define methodologies to reduce the system failures. The results were obtained within the partnership among the High Voltage Laboratory of the Federal University of Itajub\ue1, AES Sul Utility Company and the University of Bologna. Direct discharges and induced surges were simulated into real networks to identify the major factor of influence for network failures. Then commentaries on the relative performance and comparisons of different construction configurations of overhead lines are presented. Once that the atmospheric discharges phenomena are random, this work considers that the parameter generation of the discharges follows the statistical data proposed by Anderson and Eriksson. The Monte Carlo Method is used for the incidence distribution of the discharges and the Electro Geometrical Model for the interception point of the discharge

Double inhibition of cAMP and mTOR signalling may potentiate the reduction of cell growth in ADPKD cells

BACKGROUND: ADPKD is a renal pathology caused by mutations of PKD1 and PKD2 genes, which encode for polycystin-1 (PC1) and polycystin-2 (PC2), respectively. PC1 plays an important role regulating several signal transducers, including cAMP and mTOR, which are involved in abnormal cell proliferation of ADPKD cells leading to the development and expansion of kidney cysts that are a typical hallmark of this disease. Therefore, the inhibition of both pathways could potentiate the reduction of cell proliferation enhancing benefits for ADPKD patients. METHODS: The inhibition of cAMP- and mTOR-related signalling was performed by Cl-IB-MECA, an agonist of A3 receptors, and rapamycin, respectively. Protein kinase activity was evaluated by immunoblot and cell growth was analyzed by direct cell counting. RESULTS: The activation of A3AR by the specific agonist Cl-IB-MECA causes a marked reduction of CREB, mTOR, and ERK phosphorylation in kidney tissues of Pkd1 flox/-: Ksp-Cre polycystic mice and reduces cell growth in ADPKD cell lines, but not affects the kidney weight. The combined sequential treatment with rapamycin and Cl-IB-MECA in ADPKD cells potentiates the reduction of cell proliferation compared with the individual compound by the inhibition of CREB, mTOR, and ERK kinase activity. Conversely, the simultaneous application of these drugs counteracts their effect on cell growth, because the inhibition of ERK kinase activity is lost. CONCLUSION: The double treatment with rapamycin and Cl-IB-MECA may have synergistic effects on the inhibition of cell proliferation in ADPKD cells suggesting that combined therapies could improve renal function in ADPKD patients

Clinical and epidemiological features of blood stream infection caused by Candida "psilosis" complex

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