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Intensity modulated radiation therapy (IMRT) is increasingly employed in glioblastoma (GBM) treatment. The present work aimed to assess which clinical-dosimetric scenario could benefit the most from IMRT application, with respect to three-dimensional conformal radiation therapy (3D-CRT). The number of organs at risk (OARs) overlapping the planning target volume (PTV) was the parameter describing the clinical-dosimetric pattern. Based on the results, a dosimetric decision criterion to select the most appropriate treatment technique is provided. Seventeen previously irradiated patients were retrieved and re-planned with both 3D-CRT and IMRT. The prescribed dose was 60 Gy/30fx. The cases were divided into 4 groups (4 patients in each group). Each group represents the scenario where 0, 1, 2 or 3 OARs overlapped the target volume, respectively. Furthermore, in one case, 4 OARs overlapped the PTV. The techniques were compared also in terms of irradiated healthy brain tissue. The results were evaluated by paired t-test. IMRT always provided better target coverage (V95%) than 3D-CRT, regardless the clinical-dosimetric scenario: difference ranged from 0.82% (p= 0.4) for scenario 0 to 7.8% (p= 0.02) for scenario 3, passing through 2.54% (p= 0.18) and 5.93% (p= 0.08) for scenario 1 and 2, respectively. IMRT and 3D-CRT achieved comparable results in terms of dose homogeneity and conformity. Concerning the irradiation of serial-kind OARs, both techniques provided nearly identical results. A statistically significant dose reduction to the healthy brain in favor of IMRT was scored. IMRT seems a superior technique compared to 3D-CRT when there are multiple overlaps between OAR and PTV. In this scenario, IMRT allows for a better target coverage while maintaining equivalent OARs sparing and reducing healthy brain irradiation. The results from our patients dataset suggests that the overlap of three OARs can be used as a dosimetric criterion to select which patients should receive IMRT treatment.

Hypertrophic cardiomyopathy (HCM) is the most frequent autosomal dominant genetic heart muscle disease and the most common cause of sudden cardiac death in young people (under 30 y of age), who are often unaware of their underlying condition. Genetic screening is now considered a fundamental tool for clinical management of HCM families. However, the high genetic heterogeneity of HCM makes genetic screening very expensive. Here, we propose a new high-throughput genotyping method based on a HCM 96-well sequencing plate for the analysis of 8 of the most frequent HCM-causing sarcomeric genes by automating several processes required for direct sequencing, using a commercially available robotic systems and routinely used instruments. To assess the efficiency of the robot-assisted method, we have analyzed the entire coding sequence and flanking intronic sequences of the 8 sarcomeric genes in samples from 18 patients affected by HCM and their relatives, which revealed 9 different mutations, 3 of which were novel. The automated, robot-assisted assembling of polymerase chain reaction, purification of polymerase chain reaction products, and assembly of sequencing reactions resulted in a substantial saving of time, reagent costs, and reduction of human errors, and can therefore be proposed as a primary strategy for mutation identification in HCM genetic screening in many medical genetic laboratories.

A series of natural amino acid (alanine, valine, phenylalanine, and isoleucine) amides have been synthesized and fully characterized. They have been used as supporting ligands in the Ru(II)-catalyzed asymmetric transfer hydrogenation (ath) of acetophenone in the presence of i-PrOH/KOH. Secondary amides impart high reactivity to the corresponding Ru(II) complexes, with TOFs up to 1680 h^-1 and ee's up to 47%. The amino acid and the substituents of the amide nitrogen govern the activity and the enantioselectivity of the catalytic processes. The precatalysts obtained by reacting (l)-phenylalanine p-anisidineamide and (l)-valine o-anisidineamide with [Ru(p-cymene)Cl2]2 have been isolated and characterized as the half-sandwich complexes [(η⁶-p-cymene)Ru(κ²-N,N‘-aminoamidato)Cl] (10 and 11, respectively). An ESI-MS study conducted on the acetophenone reduction catalyzed by 11 has led to the detection of the organometallic intermediates involved in the catalytic cycle:  the precatalyst 11, the 16e^- complex [(η⁶-p-cymene)Ru(κ²-N,N‘-diamide)] 12, and the hydride [(η⁶-p-cymene)Ru(κ²-N,N‘-aminoamidato)H] 13.

Thirty-six samples, including fecal specimens, dry feedstuffs, raw and processed pork meat products, and dry fermented sausages, were collected from two production chains of swine meat commodities and analyzed for the presence of 11 antibiotic resistance (AR) genes. Specific PCR assays carried out on DNA extracted directly from the samples revealed a high incidence of the genes tet(K)(80.5%), ermB(66.7%), and tet(M)(66.7%). Feces and feedstuffs gave the largest number of positive amplifications. To elucidate the contribution of enterococci to the occurrence and spread of AR, 146 resistant enterococci were isolated, and their identity, genetic fingerprints, and AR gene profiles were determined by means of molecular techniques. Enterococcus faecalisand Enterococcus faeciumwere the predominant isolated species (43.8 and 38.4%, respectively); other Enterococcusspecies identified were E. durans(8.9%), E. hirae(2.7%), E. gallinarum(2.1%), E. mundtii(2.1%), and E. casseliflavus(2.1%). A number of isolates displayed a complex AR gene profile comprising up to four different resistance determinants. The genes tet(M)and ermBwere highly diffused, being present in 86.9 and 84.9%, respectively, of the isolates. The application of amplified fragment length polymorphism fingerprinting was particularly valuable to monitor the resistant enterococcal isolates along the production chain and to individuate steps in which contamination might occur. In fact, isolates of E. faecalisand E. faeciumshowing the same amplified fragment length polymorphism profile and AR gene pattern were detected in samples taken at different steps of the food chain suggesting three cases of bacterial clonal spread.

AbstractThe use of highly sensitive immunometric methods in clinical laboratories to assay anti-thyroid antibodies has progressively expanded in recent years but it is not known whether the new techniques have improved the analytical variability connected with the preceding methodologies. The Italian Society of Laboratory Medicine Study Group on Autoimmune Diseases conducted a collaborative study with the biomedical industry to evaluate the degree of standardization of the new analytical procedures. Twelve companies agreed to participate in the study on the search for anti-thyroglobulin (anti-Tg) and anti-thyroperoxidase (anti-TPO) antibodies in nine sera from patients with autoimmune thyroiditis, and in six sera from patients with non-autoimmune thyroid disease; ten immunometric and three immunofluorescence methods were employed.Agreement of qualitative results was close to 90% for anti-Tg and 97% for anti-TPO, with no important differences between the methods; variability of the quantitative results, expressed as CV% of absolute (in IU/ml) and relative (in cut-off concentration multiples) values was 93.9% and 102.3%, respectively, for anti-Tg, and 75.5% and 62.9%, respectively, for anti-TPO. These findings show that despite the progressive improvement in the analytical techniques, the variability between methods for the assay of anti-Tg and anti-TPO is still unexpectedly high, and probably due to several factors such as uncertainty in defining the positive cutoff concentration, absence of adequate international reference preparations, modality of autoantigen purification, and analytical variability in the assay procedures.

The influence of laparoscopic cholecystectomy (LC) on the mechanical properties of the respiratory system (RS) was examined using multiple regression analysis (MRA). Measurements of airway pressure (PaO) and flow (V′) were obtained from 32 patients at four distinct stages of the LC procedure: 1) Immediately before the application of pneumoperitoneum (PP) at supine position, 2) 5 min after the induction of PP at Trendelenburg position, 3) 5 min after the patient's position at reverse Trendelenburg, and 4) 5 min after the end of the surgical procedure with the patient again in supine position. Evaluated parameters were the RS elastance (Ers), resistance (Rrs), impedance (Zrs), the angle ϕindicating the balance between the elastic and resistive components of the impedance, as well as the end-expiratory elastic recoil pressure (EEP). Ers and Zrs increased considerably during PP and remained elevated immediately after abolishing PP. Rrs, on the contrary, returned to pre-operative levels right after the operation. Change of body position from Trendelenburg (T) to reverse Trendelenburg (rT) mainly induced a significant change in ϕ, thus indicating an increased dominance of the elastic component of Zrs on changing from T to rT. There was no evidence of increased End-Expiratory Pressure during PP.