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Medical records are a relevant source for real-world evidence. We introduced patient-reported outcomes (PROs) in clinical practice, demonstrating a significant quality-of-life improvement, compared to usual visit. In this secondary analysis, we describe the agreement between patients' and physicians' reports of 5 symptoms. Our hypothesis was that adoption of PROs questionnaire could significantly improve the agreement.Eligible patients were receiving active anti-cancer treatment. Patients in the control group underwent usual visits (group A), while patients of group B, before each visit, filled a PROs paper questionnaire, to provide information about symptoms and toxicities. No specific instructions were provided to physicians to integrate such information in medical records. Agreement between patient and physician evaluations was assessed by Cohen's κ, calculating under-reporting as proportion of toxicities reported by patients but not recorded by physicians.211 patients (412 visits) have been analyzed. For all symptoms, Cohen's κ was better for group B: emesis (0.25 group A vs. 0.36 group B), diarrhea (0.16 vs. 0.57), constipation (0.07 vs. 0.28), pain (0.22 vs. 0.42), fatigue (0.03 vs. 0.08). For all symptoms, although under-reporting was relevant in both groups, it was lower for group B: emesis (75.49% vs. 60.0%, p=0.031), diarrhea (82.89% vs. 50.0%, p0.001), constipation (92.11% vs. 69.57%, p0.001), pain (59.57% vs. 42.31%, p=0.01), fatigue (82.11% vs. 64.10%, p0.001).Adoption of paper PROs allowed a significant reduction in under-reporting of symptoms, but agreement remained suboptimal. Direct integration of electronic PROs could minimize the issue of under-reporting of medical records, increasing their accuracy.

In this study we exposed 1,2-dioleoyl-3-trimethylammonium propane (DOTAP)/DNA lipoplexes to different concentrations of human plasma (HP). Liquid chromatography-tandem mass spectrometry showed that the adsorbed protein corona is rich in vitronectin, a major ligand for α vβ 3 or α vβ 5 integrins. To investigate whether the protein corona could dictate a selective access to cells expressing vitronectin receptors, highly-metastatic MDA-MB-435S cells that express high levels of α vβ 3 and α vβ 5 integrins, were used as experimental model. Non-metastatic MCF7 cells that do not express α vβ 3 and express reduced levels of α vβ 5 integrins were used as a reference. After exposure to HP, the cellular uptake of lipoplexes in MDA-MB-435S cells was more than 2-fold larger than that of bare ones, while complexes could not enter MCF7 cells remaining largely accumulated at the plasma membrane. Collectively our data suggest that the protein corona can be used as a novel tool for cancer cell targeting.

Most lipid formulations need cholesterol for efficient transfection, but the precise motivation remains unclear. Here, we have investigated the effect of cholesterol on the transfection efficiency (TE) of cationic liposomes made of l, 2-dioleoyl-3-trimethylammonium-propane (DOTAP) and dioleoylphosphocholine (DOPC) in Chinese hamster ovary cells. The transfection mechanisms of cholesterol-containing lipoplexes have been investigated by TE, synchrotron small angle X-ray scattering (SAXS) and laser scanning confocal microscopy (LSCM) experiments. We prove that cholesterol-containing lipoplexes enter the cells using different endocytosis pathways. Formulations with high cholesterol content efficienlty escape from endosomes. These studies will contribute to the rational design of novel lipid nanocarriers with superior TE.

The authors examined relationships between quantified EEG (qEEG) variables and neuropsychological performance in 54 consecutive patients with probable Alzheimer's disease (AD). Patients were studied with qEEG and a neuropsychological battery that assessed memory, attention, verbal functions, set-shifting abilities, and procedural learning. More severe memory, attention, and verbal deficits were significantly correlated with lower alpha relative power, and increased theta relative power was significantly correlated with poor set-shifting abilities. No qEEG variables were significantly correlated with the procedural learning task. These correlations between deficits in specific cognitive domains and qEEG bands suggest that qEEG may have an important role in the investigation of the cognitive deficits in AD.

We examined the presence of quantified electroencephalographic (qEEG) differences between Parkinson's disease (PD) patients with and without dementia, and a group of age-comparable normal controls. While there were no significant differences in relative power in any of the qEEG bands between PD patients without dementia and normal controls, PD patients showed a significantly greater reactivity in the alpha band. On the other hand, PD patients with dementia showed significantly less alpha and more theta relative power than both the normal control and the PD without dementia groups. When PD patients with dementia were compared with a group of patients with Alzheimer's disease (AD) and a similar severity of dementia, no significant between-group differences were found in any of the qEEG bands. In conclusion, while our study demonstrated no significant qEEG differences between non-demented PD patients and normal controls, PD patients with dementia showed qEEG changes similar to those observed in patients with AD.

Safe and efficient intracellular delivery of genes or drugs is critically important in targeted cancer treatment and gene therapy applications. Ultrasound (US) has been demonstrated to alter the cell membrane permeability due to a biophysical mechanism (Sonoporation) and exploited as a promising non-invasive gene transfer method. The sonoporation process could induce the formation of transient pores without significantly affecting cell viability. This research is aimed at investigating some bioeffects due to Therapeutic Ultrasound (pulsed-1 MHz) which could allow to enhance drugs or genes delivery in a non tumoral cell line. We have used the NIH-3T3 cell line as model system and exposed it to US at two different distances from the source; the effects of this pulsed ultrasonic wave on cells were assessed by Fourier transform infrared (FT-IR) spectroscopic imaging analysis. This technique combined with a focal plane array (FPA) detector has been widely used to study the general biochemical changes in vitro; moreover, the development of FPA detectors and shortening of measurement times specifically for IR imaging, from several hours to few minutes, have made possible to image the distribution of molecular species in biological samples. We have also performed a cytokinesis-block micronucleus (CBMN) assay to reveal the presence or not of micronuclei (named Howell-Jolly bodies) formed during the cell division due to the DNA damage. The results of IR analysis combined with the cytogenetic analysis have shown that these experimental conditions can not cause DNA mutations in the NIH-3T3 cell line. Finally, the comparison between the spectral parameters of the average spectrum extracted from the spectral map and those of the set of all spectra from the spectral map could be limited by the presence of bad pixels inside the map.

We used Fourier Transibrm Infrared Spectroscopy (FTIR) combined with flow cytometry to study the apoptosis and necrosis processes in Jurkat, a lymphocyte cell lìne. The apoptosis was induced in the cells by a chemical agent, the actinomycin D, while the necrosis was induced lowering the pH value to 4.2. The apoptotic events were analysed by flow cytometry (using annexin V and propidium iodide) and contemporary monitored by FTIR spectroscopy at diflèrent times after the treatment. This comparison allowed us to find in the IR spectrum, between 3000 cm 1 and 2800 c--1, a "marker band" of the apoptosis corresponding to the exposure of phosphatidylserine on the outer leaflet of the membrane. A marker of a specific cellular process obtained by using a non-destructive technique such as FTIR spectroscopy, has a great signifìcance in the diagnostic medicine providing a tool for detecting pathologies in vivo.

In the present study, we investigated the effect of the interaction of electromagnetic radiation with human keratinocytes (HaCaT)at low frequency. HaCaT cells were exposed to a sinusoidal magnetic field at a density of 2 mT (50 Hz)These cells are a good model for studying interaction of non-ionising radiation, because they are totally open to electromagnetic waves and also because they are resistant to both mechanical and thermal stimuli. We performed scanning microscopy which showed modification in shape and morphology in exposed cells. This modification is related to differential actin distribution as revealed by phalloidin fluorescence analysis. Moreover, 50 Hz exposed cells show increased clonogenic capacity, as well as increased cellular growth as showed by clonogenicity assays and growth curves. Indirect immunofluorescence analysis using a fluorescent antibody against involucrin and b4 integrin differentiation and adhesion markers respectively, revealed an increase of involucrin expression and segregation of b4 integrin in the cell membrane in 50 Hz exposed cells, suggesting that a higher percentage of the exposed cells shows a modified pattern of adhesion and differentiation markers. We also present evidence that electromagnetic field (EMF) exposure of HaCaT cells can interfere with protein kinase activity. Our observations confirm the hypothesis that electromagnetic fields, at 50 Hz may modify cell membrane morphology and interfere with initiation of the signal cascade pathway, and cellular adhesion.

In this work we employed Atomic Force Microscopy (AFM) and Scanning Near-field Optical Microscopy (SNOM) techniques to study the effect of the interaction between human keratinocytes (HaCaT) and electromagnetic fields at low frequency. HaCaT cells were exposed to a sinusoidal magnetic field at a density of 50 Hz, 1mT. We performed AFM analysis showed modification in shape and morphology in exposed cells with an increase of the areas of adhesion between cells. This last evidence has been confirmed by SNOM indirect immunofluorescence analysis performed with a fluorescent antibody against the adhesion marker b4 integrin, which revealed an increase of b4 integrin segregation in the cell membrane of 50 Hz exposed cells, suggesting that a higher percentage of these cells shows a modified pattern of this adhesion marker

Air operating atomic force microscopy has been used to study human B lymphoblastoid (Raji) cells exposed to a 50 Hz 2 mT sinusoidal magnetic field (MF), up to 64 h. A statistical analysis performed on mean height and membrane roughness of the cells showed a loss of microvilli and several modifications of the overall shape and membrane surface at

Air operating atomic force microscopy (AFM) has been used to study human B lymphoblastoid (Raji) cells exposed to a 50 Hz 2mT sinusoidal magnetic field (MF), up to 64 hours. A statistical analysis performed on mean height and membrane roughness of the cells showed a loss of microvilli and several modifications of the overall shape and membrane surface at increasing MF exposure.

The authors examined the presence of specific quantified electroencephalographic (qEEG) changes in dementia patients with relatively lower frontal or parietal blood perfusion as demonstrated by SPECT. Over all brain regions, patients with relatively lower parietal perfusion showed significantly higher theta relative power than demented patients with relatively lower frontal perfusion or normal control subjects. Dementia patients with relatively lower frontal perfusion showed no differences from age-comparable normal control subjects in qEEG variables. These findings 1) suggest that usefulness of qEEG for the diagnosis of dementia is restricted to a subgroup of patients with the typical SPECT pattern of parietal blood hypoperfusion and 2) demonstrate that the qEEG changes typical of dementia are not related to perfusion deficits in frontal brain areas.

Fusion of Epstein-Barr virus (EBV) with Raji cells was measured after exposure of the virus to neutral or low pH, enzymatic modification of the viral spike glycoproteins, or chemical modification of the target membrane. The relief of octadecylrhodamine (R18) fluorescence self-quenching was used to monitor fusion. Fusion of EBV with Raji cells at pH 5.9 was significantly enhanced compared to that at neutral pH. Treatment of Raji cells with agents known to modify the surface net charge (trinitrobenzene sulfonic acid) totally prevented fusion at a neutral pH. Desialylation of EBV significantly reduced the extent of fusion with Raji cells. Our results demonstrate that EBV is rapidly internalized and then fuses with lymphoblastoid cells in the endocytic vesicles.

When nanoparticles (NPs) are exposed to biological fluids they adsorb biomolecules (mainly proteins) to form a protein corona. We have applied nanoliquid-cromatography mass spectroscopy (NanoLC-MS/MS) to quantitatively determine the proteins associated with several lipid NP formulations after incubation with human plasma (HP). By NanoLC-MS/MS we identified vitronectin as the most promising protein corona component for active targeting. To exploit the "protein corona effect" we investigated the interactions of l, 2-dioleoyl-3-trimethylammonium propane (DOTAP)/DNA cationic liposome/DNA complexes (lipoplexes) with target cells. By applying two-color fluorescence laser scanning confocal microscopy (LSCM) we demonstrate that vitronectin directs a receptor-mediated uptake of lipoplexes into target cells. This experiment sets the basis for a rational exploitation of the protein corona for targeted drug and gene delivery.

The EBV genome in latently infected lymphoid cells offers an opportunity to follow effects on the transcriptional and translational product clearly distinguishable from those of the host cell genome. Exposure of Akata cells, a human lymphoid cell line latently infected by the EBV genome, to a 50 Hz EMF resulted in an increased number of cells expressing the virus early antigens. This finding provides additional evidence that DNA can be modulated by a magnetic field.