Immune biology of NSCLC revealed by single-cell technologies: implications for the development of biomarkers in patients treated with immunotherapy.

First-line immunotherapy in non-small-cell lung cancer largely improved patients' survival. PD-L1 testing is required before immune checkpoint inhibitor initiation. However, this biomarker fails to accurately predict patients' response. On the other hand, immunotherapy exposes patients to immune-related toxicity, the mechanisms of which are still unclear. Hence, there is an unmet need to develop clinically approved predictive biomarkers to better select patients who will benefit the most from immune checkpoint inhibitors and improve risk management. Single-cell technologies provide unprecedented insight into the tumor and its microenvironment, leading to the discovery of immune cells involved in immune checkpoint inhibitor response or toxicity. In this review, we will underscore the potential of the single-cell approach to identify candidate biomarkers improving non-small-cell lung cancer patients' care.

4He dose- and track-averaged linear energy transfer: Monte Carlo algorithms and experimental verification.

Objective.In the present hadrontherapy scenario, there is a growing interest in exploring the capabilities of different ion species other than protons and carbons. The possibility of using different ions paves the way for new radiotherapy approaches, such as the multi-ions treatment, where radiation could vary according to target volume, shape, depth and histologic characteristics of the tumor. For these reasons, in this paper, the study and understanding of biological-relevant quantities was extended for the case of4He ion.Approach.Geant4 Monte Carlo based algorithms for dose- and track-averaged LET (Linear Energy Transfer) calculations, were validated for4He ions and for the case of a mixed field characterised by the presence of secondary ions from both target and projectile fragmentation. The simulated dose and track averaged LETs were compared with the corresponding dose and frequency mean values of the lineal energy,yD¯andy¯F, derived from experimental microdosimetric spectra. Two microdosimetric experimental campaigns were carried out at the Italian eye proton therapy facility of the Laboratori Nazionali del Sud of Istituto Nazionale di Fisica Nucleare (INFN-LNS, Catania, I) using two different microdosimeters: the MicroPlus probe and the nano-TEPC (Tissue Equivalent Proportional Counter).Main results.A good agreement ofL¯dTotalandL¯tTotalwithy¯Dandy¯Texperimentally measured with both microdosimetric detectors MicroPlus and nano-TEPC in two configurations: full energy and modulated4He ion beam, was found.Significance.The results of this study certify the use of a very effective tool for the precise calculation of LET, given by a Monte Carlo approach which has the advantage of allowing detailed simulation and tracking of nuclear interactions, even in complex clinical scenarios.

Evidence of Vasculogenic Mimicry in a Palpebral Melanocytoma in a Dog.

A 7-year-old neutered female Doberman pinscher was presented with a palpebral nodule on the haired eyelid of the left eye. The nodule was removed surgically. Microscopically, the nodule was consistent with eyelid melanocytoma. The tumour was characterized by the presence of numerous lacunar and slit-like spaces filled by erythrocytes and interspersed throughout the neoplastic melanocytes. Immunohistochemically, these spaces were lined by cells expressing PNL2, but the cells were negative for factor VIII and CD31. [corrected] These findings were consistent with neoplastic melanocytes without endothelial cell participation. This feature was interpreted as 'vasculogenic mimicry', a mechanism of tumour angiogenesis that is well-recognized in human melanomas, but has not yet been reported in melanomas in animals.

Search for Event Rate Modulation in XENON100 Electronic Recoil Data.

We have searched for periodic variations of the electronic recoil event rate in the (2-6) keV energy range recorded between February 2011 and March 2012 with the XENON100 detector, adding up to 224.6 live days in total. Following a detailed study to establish the stability of the detector and its background contributions during this run, we performed an unbinned profile likelihood analysis to identify any periodicity up to 500 days. We find a global significance of less than 1σ for all periods, suggesting no statistically significant modulation in the data. While the local significance for an annual modulation is 2.8σ, the analysis of a multiple-scatter control sample and the phase of the modulation disfavor a dark matter interpretation. The DAMA/LIBRA annual modulation interpreted as a dark matter signature with axial-vector coupling of weakly interacting massive particles to electrons is excluded at 4.8σ.

Management of ischemic diabetic foot.

Diabetic foot pathology represent the more disabling complication of diabetes. More the 1 million of diabetes patients undergo a lower limb amputation per year; 85% of these amputation are preceded by un ulcer that can be avoided by a prevention program. Critical limb ischemia (CLI), the only independent cause of major amputation in diabetic population, can be correctly treated when an early diagnosis is made. Both endoluminal and surgical revascularization procedures can be applied in diabetes with high rate of success when performed by skilled operator. Infection of diabetic foot, in particular in patients suffering from peripheral artery disease (PVD), may rapidly evolves in severe local or systemic infection putting the patient at high risk of major amputation or death. Together with an early diagnosis of infection and ischemia it is mandatory to apply a correct medical and surgical treatment protocol with the aim to control infection and to improve blood perfusion to the foot. In case of infection surgical procedure should be applied first while revascularization procedure will follow soonest. Antibiotic therapy should be chosen considering different local biological pattern and different type of infection. Reconstructive surgery, the last step in treatment of any diabetic foot lesion, must obtain a functional residual foot or a stump that will allow the patient to go back walking soonest with residual good walking capacity.

Limits on spin-dependent WIMP-nucleon cross sections from 225 live days of XENON100 data.

We present new experimental constraints on the elastic, spin-dependent WIMP-nucleon cross section using recent data from the XENON100 experiment, operated in the Laboratori Nazionali del Gran Sasso in Italy. An analysis of 224.6 live days×34 kg of exposure acquired during 2011 and 2012 revealed no excess signal due to axial-vector WIMP interactions with 129Xe and 131Xe nuclei. This leads to the most stringent upper limits on WIMP-neutron cross sections for WIMP masses above 6 GeV/c², with a minimum cross section of 3.5×10(-40) cm² at a WIMP mass of 45 GeV/c², at 90% confidence level.

Dark matter results from 225 live days of XENON100 data.

We report on a search for particle dark matter with the XENON100 experiment, operated at the Laboratori Nazionali del Gran Sasso for 13 months during 2011 and 2012. XENON100 features an ultralow electromagnetic background of (5.3 ± 0.6) × 10(-3) events/(keV(ee) × kg × day) in the energy region of interest. A blind analysis of 224.6 live days × 34 kg exposure has yielded no evidence for dark matter interactions. The two candidate events observed in the predefined nuclear recoil energy range of 6.6-30.5 keV(nr) are consistent with the background expectation of (1.0 ± 0.2) events. A profile likelihood analysis using a 6.6-43.3 keV(nr) energy range sets the most stringent limit on the spin-independent elastic weakly interacting massive particle-nucleon scattering cross section for weakly interacting massive particle masses above 8 GeV/c(2), with a minimum of 2 × 10(-45) cm(2) at 55 GeV/c(2) and 90% confidence level.

Dark matter results from 100 live days of XENON100 data.

We present results from the direct search for dark matter with the XENON100 detector, installed underground at the Laboratori Nazionali del Gran Sasso of INFN, Italy. XENON100 is a two-phase time-projection chamber with a 62 kg liquid xenon target. Interaction vertex reconstruction in three dimensions with millimeter precision allows the selection of only the innermost 48 kg as the ultralow background fiducial target. In 100.9 live days of data, acquired between January and June 2010, no evidence for dark matter is found. Three candidate events were observed in the signal region with an expected background of (1.8 ± 0.6) events. This leads to the most stringent limit on dark matter interactions today, excluding spin-independent elastic weakly interacting massive particle (WIMP) nucleon scattering cross sections above 7.0 × 10(-45) cm(2) for a WIMP mass of 50 GeV/c(2) at 90% confidence level.

First dark matter results from the XENON100 experiment.

The XENON100 experiment, in operation at the Laboratori Nazionali del Gran Sasso in Italy, is designed to search for dark matter weakly interacting massive particles (WIMPs) scattering off 62 kg of liquid xenon in an ultralow background dual-phase time projection chamber. In this Letter, we present first dark matter results from the analysis of 11.17 live days of nonblind data, acquired in October and November 2009. In the selected fiducial target of 40 kg, and within the predefined signal region, we observe no events and hence exclude spin-independent WIMP-nucleon elastic scattering cross sections above 3.4 × 10⁻44 cm² for 55 GeV/c² WIMPs at 90% confidence level. Below 20 GeV/c², this result constrains the interpretation of the CoGeNT and DAMA signals as being due to spin-independent, elastic, light mass WIMP interactions.

Neck lesions mimicking thyroid pathology.

INTRODUCTION: Nodular lesions within the neck may origin from several structures. A misdiagnosed origin may expose the surgeon to inappropriate procedures. These lesions are paradoxically frequent in high specialised centre for endocrine surgery. PATIENTS AND METHODS: In the year 2006, three patients were first admitted to our department with a diagnosis of thyroid nodule (1) or lymphatic metastases of thyroid carcinoma (2). The first patient had ultrasound (US) and Tc-99-m scan orienting for thyroid nodule. The two other patients, presented with lateral neck lesion in ipsilateral sincronous and previous diagnosis of papillary thyroid carcinoma, respectively, with US and computed tomography scan confirmed lesion but with a FNA cytology negative for tumoural cells. RESULTS: All three patients underwent surgical exploration. In the first two cases, a whitish tender nodule (4 and 4.5cm), cleavable from surrounding structures, was removed with final histology of Schwannoma and Paraganglioma, respectively. Both patients experienced Bernard Horner Syndrome. In the last patients, a firm grey nodule of 5cm strictly adherent to muscular planes was removed with diagnosis of Castleman's Disease. CONCLUSIONS: Nodular neck lesions mimicking a thyroid pathology (thyroid nodules or metastatic lymph nodes) are rare but can represent a tough challenge for surgeons who might fall into incorrect surgical approaches, resulting in high morbidity. Pre-operative work-up would help the surgeon to obtain the correct diagnosis, thus, to follow the better surgical approach. Nevertheless, a careful approach would be used for that neurogenic tumour amenable of resection without jeopardising nervous structures.

Gas chromatography-mass spectrometry measurement of 6beta-OH-cortisol/cortisol ratio in human urine: a specific marker of enzymatic induction.

The urinary 6beta-OH-cortisol/cortisol ratio is a specific, non-invasive marker for evaluating inductive or inhibitory effects on cytochrome P450 3A activity. We propose a new quantitative gas chromatography-mass spectrometry with isotope dilution (GC-ID-MS) method for the simultaneous determination of urinary free cortisol (UFC) and 6beta-OH-cortisol (6beta-OHC). The method utilizes the following: (a) addition of internal standard (2H2 cortisol) to 1 ml of urine; (b) loading on to an Extrelut column and elution with dichloromethane; (c) derivatization to dimethoxime tri-(trimethyl-silyl)ether (MOX-TMS); (d) separation and identification by GC-ID-MS. The detection limit for cortisol was 22 pg injected (signal-to-noise ratio 10:1) and for 6beta-OH-cortisol 123 pg injected (signal-to-noise ratio 10:1). The intra-assay and the inter-assay imprecision were 4.69% and 7.4% for 6beta-OHC and 2.44% and 3.53% for cortisol, respectively. We used this method to analyze 57 morning urine samples of healthy volunteers and patients under different conditions. We found that chronic alcoholics had a significantly higher ratio of 6beta-OHC/UFC compared to controls (p<0.0001), whereas adults undergoing methadone therapy and patients with acute alcohol intoxication exhibited a significantly lower urinary 6beta-OHC/UFC ratio (p<0.05 and p<0.01, respectively). The proposed method allows a rapid and accurate assessment of the 6beta-OHC/UFC ratio.

[Sentinel lymph nodes in skin melanoma]. / Il linfonodo sentinella nel melanoma cutaneo.

BACKGROUND: Early node dissection offers increased survival in patients with node metastases only. The study of sentinel node (SN) using blue dye and radiolocalization permits to identify patients who could undergo lymphadenectomy. METHODS: At Department of Surgery of Macerata General Hospital 22 patients with melanoma of trunk or limbs at I and II stage were submitted to SN biopsy. RESULTS: SN was identified in all cases by combined approach. Dynamic lymphoscintigraphy permitted identification of SL when it was not the node nearest the tumor. Nodal metastases were found in 2 cases (9.1%) and the patients underwent regional lymphadenectomy. In 1 patient the SN was the only node with metastases. Both patients had high thickness melanomas. The measurement of radioactive exposition for operating room personnel and pathologist demonstrated that the technique is safe and without risks. CONCLUSIONS: Biopsy of SN is effective for identification of occult nodal metastases from cutaneous melanoma. Combined technique allows to localize SN in 100% of cases. There is not radio-exposition for operators.

[The value of fine-needle biopsy under ultrasonic control in the diagnosis of kidney tumors]. / Die Wertigkeit der ultraschallgeleiteten Feinnadelpunktion in der Diagnostik von Nierentumoren.

127 fine-needle aspiration biopsies obtained under ultrasonographic control from circumscribed kidney lesions of 102 patients (with histologically examined lesions) were selected from a total of 867 fine-needle aspiration biopsies performed in the years between 1978 and 1986. The result of the cytological examination was compared with the histological diagnosis of the lesion. A correct diagnosis of the nature of the lesion (concerning dignity) was achieved in 90.2% of the cases under optimal circumstances. No mistakenly positive diagnosis was made. A total of 88% of the histologically malignant tumors or 80.2% of the kidney cancers were correctly diagnosed already on the basis of the cytological examination. The fine-needle aspiration can be repeated when the obtained material is insufficient: A kidney cancer was diagnosed in this way 13 times among 22 patients. The cytological examination permitted a more precise tumor-type differentiation in the large majority of cases. These results demonstrate the value of this diagnostic tool that carries only an irrelevant risk.