The presence of visual gap affects the duration of stopping process

A milestone on which relies the voluntary control of behavior is the ability to shape our motor output to meet the needs of the context which we are continuously facing. Even though it is solidly established that contextual information influence movement generation few studies have so far explored their effects on inhibitory processes. We compared the inhibitory control of arm movements of ten healthy right-handed volunteers in a countermanding reaching paradigm with and without the presence of a temporal gap between the offset of the central target and the peripheral target appearance. We found that this perceptual gap reduces the reaction times of hand movements and, at the same time, increases the duration of the stop process, the stop signal reaction time. The two effects are not correlated implying that inhibition and execution of reaching movement are two independent processes influenced by a common factor: the disengagement of selective attention from the central target. Therefore our results support the idea of the existence of a link between spatial selective attention and inhibitory processes. G. Mirabella and P. Pani equally contributed to the present paper.     

Urinary tract reconstruction using the controlateral native ureter and a combined open-retroperitoneoscopic approach after renal transplantation

An alternative technique for urinary tract (UT) reconstruction is described in a renal transplant recipient who developed a severe stenosis of the graft ureter. This approach entails the retroperitoneoscopic preparation of the native ureter contralateral to the graft, followed by an open reconstruction of the UT. The ureter was dissected along its entire length to the level of the iliac vessels, with its associated mesentery still attached in order to preserve the vascular supply. The corresponding native kidney contralateral to the graft was endoscopically removed. A longitudinal sub-umbilical incision allowed the excision of the stenotic tract and the reconstruction of the UT by means of a manual end-to-end anastomosis between the new ureter and the graft pelvis. No post-operative complications occurred and renal function immediately resumed. The approach described represents an alternative solution for the surgical management of severe ureteric graft stenosis. We believe that the magnification of the anatomy granted by the endoscope during the dissection of the ureter and neighboring structures provides the gentle handling of the tissues and the remote dissection away from the ureter with the highest precision.     

Progression patterns and therapeutic sequencing following immune checkpoint inhibition for hepatocellular carcinoma: An international observational study

Background and Aims

Different approaches are available after the progression of disease (PD) to immune checkpoint inhibitors (ICIs) for hepatocellular carcinoma (HCC), including the continuation of ICI, treatment switching to tyrosine kinase inhibitors (TKIs) and cessation of anticancer therapy. We sought to characterise the relationship between radiological patterns of progression and survival post-ICI, also appraising treatment strategies.

Methods

We screened 604 HCC patients treated with ICIs, including only those who experienced PD by data cut-off. We evaluated post-progression survival (PPS) according to the treatment strategy at PD and verified its relationship with radiological patterns of progression: intrahepatic growth (IHG), new intrahepatic lesion (NIH), extrahepatic growth (EHG), new extrahepatic lesion (NEH) and new vascular invasion (nVI).

Results

Of 604 patients, 364 (60.3%) experienced PD during observation. Median PPS was 5.3 months (95% CI: 4.4–6.9; 271 events). At the data cut-off, 165 patients (45%) received no post-progression anticancer therapy; 64 patients (17.6%) continued ICI beyond PD. IHG (HR 1.64 [95% CI: 1.21–2.22]; p = .0013) and nVI (HR 2.15 [95% CI: 1.38–3.35]; p = .0007) were associated with shorter PPS. Multivariate models adjusted for progression patterns, treatment line and albumin-bilirubin grade and Eastern Cooperative Oncology Group performance status at PD confirmed receipt of ICI beyond PD with (HR 0.17, 95% CI: 0.09–0.32; p < .0001) or without subsequent TKI (HR 0.39, 95% CI: 0.26–0.58; p < .0001) as predictors of prolonged PPS versus no anticancer therapy.

Conclusions

ICI-TKI sequencing is a consolidated option in advanced HCC. nVI and IHG predict a poorer prognosis. Despite lack of recommendation, the continuation of ICI beyond progression in HCC is adopted clinically: future efforts should appraise which patients benefit from this approach.     

Dispersive liquid-liquid microextraction,an effective tool for the determination of synthetic cannabinoids in oral fluid by liquid chromatography–tandem mass spectrometry

In the present work, dispersive liquid-liquid microextraction (DLLME) was used to extract six synthetic cannabinoids (JWH-018, JWH-019, JWH-073, JWH-200, or WIN 55,225, JWH-250, and AM-694) from oral fluids. A rapid baseline separation of the analytes was achieved on a bidentate octadecyl silica hydride phase (Cogent Bidentate C₁₈; 4.6 mm × 250 mm, 4 μm) maintained at 37 °C, by eluting in isocratic conditions (water:acetonitrile (25:75, V/V)). Detection was performed using positive electrospray ionization–tandem mass spectrometry. The parameters affecting DLLME (pH and ionic strength of the aqueous phase, type and volume of the extractant and dispersive solvent, vortex and centrifugation time) were optimized for maximizing yields. In particular, using 0.5 mL of oral fluid, acetonitrile (1 mL), was identified as the best option, both as a solvent to precipitate proteins and as a dispersing solvent in the DLLME procedure. To select an extraction solvent, a low transition temperature mixture (LTTM; composed of sesamol and chlorine chloride with a molar ratio of 1:3) and dichloromethane were compared; the latter (100 μL) was proved to be a better extractant, with recoveries ranging from 73% to 101 % by vortexing for 2 min. The method was validated according to the guidelines of Food and Drug Administration bioanalytical methods: intra-day and inter-day precisions ranged between 4 % and 18 % depending on the spike level and analyte; limits of detection spanned from 2 to 18 ng/mL; matrix-matched calibration curves were characterized by determination coefficients greater than 0.9914. Finally, the extraction procedure was compared with previous methods and with innovative techniques, presenting superior reliability, rapidity, simplicity, inexpensiveness, and efficiency.     

A comparison between EASI system 12-lead ECGs and standard 12-lead ECGs for improved clinical nursing practice

Aims and objectives. This study was carried out to verify the accuracy of 12‐Lead ECG, obtained through a continuous ECG monitoring system with five cables positioned in EASI mode, to identify basic ECG alterations. Background. This study concerns continuous ECG monitoring systems in Coronary Care Units. Continuous ECG monitoring is an important device for nursing surveillance and is useful in decreasing adverse events. Design and method. Thirteen patients admitted consecutively to the Coronary Care Unit for Acute Myocardial Infarction underwent daily and simultaneous recording of a12‐lead ECG using both procedures: EASI ECG and STANDARD ECG. A sample of 1164 ECG leads acquired in EASI mode was compared with a sample of as many ECG leads acquired using the standard procedure with a traditional cardiograph. Results and conclusions. In the Coronary Care Unit, Continous ECG monitoring with five cables positioned in EASI mode is a valid alternative to the standard 12‐lead ECG for cardiac rhythm abnormalities detection and for acute myocardial ischemia and old myocardial infarction assessment. Therefore, the EASI system might be advantageous for long‐term patient monitoring. Relevance to clinical practice. The EASI system represents a valid device for the nursing surveillance of patients who need continuous ECG monitoring, improves clinical nursing practice in Coronary Care Units, supports the reduction of adverse events such as cardiac arrest and reduces the hospital costs.     

Chemotherapeutic drugs may be used to enhance the killing efficacy of human tumor antigen peptide-specific CTLs

The effects of anticancer chemotherapy on antigen-specific cytotoxic T lymphocytes (CTLs) are mostly unknown. We tested the effects of cytotoxic drugs such as 5-fluorouracil, gemcitabine, and oxaliplatin on the functional activity of antigen-specific CTL cultures derived from the peripheral blood mononuclear cells of human donors. We found that a biweekly drug-exposure of human HLA-A(*)02.01+ CTLs derived from bulk cultures led to completely different effects if occurring early (day second) or late (day thirteenth) after the in vitro stimulations with the cognate peptides. In the first case, there was a significant CTL inhibition, whereas in the second, there was a marked enhancement of the antigen-specific cytolytic activity. Results of immunocytofluorimetric studies and CTL/natural killer inhibition assays suggested that the latter effect could be related to a more selective drug-mediated inhibition of cohabitant T regulatory (reg) cells. These results were translated in an in vivo therapeutic mouse model where humanized HLA-A(*)02.01 transgenic mice inoculated with EL-4/humanized HLA-A(*)02.01 transgenic mice showed a prolonged survival and the greatest rate of cure when receiving a combined treatment with a thymidylate synthase-specific peptide vaccine and a multidrug chemotherapy regimen administered late after immunization. Tumor samples derived from this group of mice showed a reduced expression of the target thymidylate synthase antigen, a marked reduction of T(reg)s, and a noteworthy infiltration of C8+ T cells. These results may have clinical implications for the design of new translational anticancer regimens aimed at combining chemotherapy and immunotherapy.     

A polyepitope DNA vaccine targeted to Her-2/ErbB-2 elicits a broad range of human and murine CTL effectors to protect against tumor challenge

A cDNA vaccine (pVax1/pet-neu) was designed to encode 12 different Her-2/ErbB-2-derived, HLA-A*0201-restricted dominant and high-affinity heteroclitic cryptic epitopes. Vaccination with pVax1/pet-neu triggered multiple and ErbB-2-specific CTL responses in HLA-A*0201 transgenic HHD mice and in HLA-A*0201 healthy donors in vitro. Human and murine CTL specific for each one of the 12 ErbB-2 peptides recognized in vitro both human and murine tumor cells overexpressing endogenous ErbB-2. Furthermore, vaccination of HHD mice with pVax1/pet-neu significantly delayed the in vivo growth of challenged ErbB-2-expressing tumor (EL4/HHD/neu murine thymoma) more actively when compared with vaccination with the empty vector (pVax1) or vehicle alone. These data indicate that the pVax1/pet-neu cDNA vaccine coding for a poly-ErbB-2 epitope is able to generate simultaneous ErbB-2-specific antitumor responses against dominant and cryptic multiple epitopes.     

Evidence for a biphasic apoptotic pathway induced by melatonin in MCF-7 breast cancer cells

Abstract:  Previous investigations demonstrated that melatonin exerts an oncostatic action on estrogen-responsive breast cancer, both in vitro and in vivo. Nevertheless, the pro-apoptotic effect of melatonin is still a matter of debate. An experimental study was undertaken to focus on melatonin-related apoptosis and to identify the apoptotic pathways involved. Whole cell-count, flow-cytometry analysis and proteins involved in apoptotic pathways [p53, p73, murine double minute 2 (MDM2), caspases-9,-7,-6, cleaved-poly ADP ribose polymerase (PARP), Bcl-2, Bax and apoptotic inducing factor (AIF)] were investigated in human MCF-7 breast cancer cells treated with physiological (1 nM) concentration of melatonin. Melatonin exerts a significant growth-inhibitory effect on MCF-7 cells, becoming evident after 72 hr and thereafter increasing linearly up to 144 hr. In this model, the growth-inhibition is transforming growth factor beta 1 (TGFβ1)-dependent and it might be reversed by adding an anti-TGFβ1 antibody. Melatonin induces a significant rise in apoptotic rate, at both 24 and 96 hr. The anti-TGFβ1 antibody almost completely suppresses melatonin-related late apoptosis; however, early apoptosis is unaffected. Early programmed cell death is associated with a significant increase in the p53/MDM2 ratio and in AIF release, without modifications in caspase activity or cleaved-PARP levels. Activated caspases-9 and -7 and cleaved-PARP increased significantly at 96 hr, concomitantly with a down-regulation of the Bcl-2/Bax ratio. These data suggest that two distinct apoptotic processes are triggered by melatonin in MCF-7 cells: an early, TGFβ1 and caspase-independent response, and a late apoptotic TGFβ1-dependent process in which activated-caspase-7 is likely to be the terminal effector.