Distinct subclonal tumour responses to therapy revealed by circulating cell-free DNA.

BACKGROUND: The application of precision medicine in oncology requires in-depth characterisation of a patient's tumours and the dynamics of their responses to treatment. PATIENTS AND METHODS: We used next-generation sequencing of circulating cell-free DNA (cfDNA) to monitor the response of a KIT p.L576P-mutant metastatic vaginal mucosal melanoma to sequential targeted, immuno- and chemotherapy. RESULTS: Despite a KIT mutation, the response to imatinib was mixed. Unfortunately, tumours were not accessible for molecular analysis. To study the mechanism underlying the mixed clinical response, we carried out whole-exome sequencing and targeted longitudinal analysis of cfDNA. This revealed two tumour subclones; one with a KIT mutation that responded to imatinib and a second KIT-wild-type subclone that did not respond to imatinib. Notably, the subclones also responded differently to immunotherapy. However, both subclones responded to carboplatin/paclitaxel, and although the KIT-wild-type subclone progressed after chemotherapy, it responded to subsequent re-administration of paclitaxel. CONCLUSION: We show that cfDNA can reveal tumour evolution and subclonal responses to therapy even when biopsies are not available.

Sublingual allergen immunotherapy in HIV-positive patients.

HIV infection is a relative contraindication for allergic immunotherapy (AIT). In the last decade, highly active antiretroviral therapy (HAART) has improved the immune function and life expectancy in HIV-infected patients whose respiratory allergic incidence is similar to the general population. We evaluated the safety and clinical effectiveness of sublingual immunotherapy in a group of grass pollen-allergic HAART-treated HIV-positive patients. Thirteen patients received sublingual immunotherapy (SLIT) tablet (Oralair, Stallergenes©) and symptomatic therapy and were compared with nine patients receiving symptomatic therapy alone. Clinical benefits were evaluated by the analysis of total combined score (TCS), sum of symptom-medication score, and a quality of life (QoL) questionnaire. HIV viral load and peripheral TCD4 lymphocytes were analyzed at the beginning and at the end of the study. Clinical efficacy data showed a significant improvement in SLIT-treated patients compared to controls (TCS: P = 0.0001; QoL: P = 0.03). We did not observe any significant alteration of TCD4 cell counts and viral load (VL) in both groups. Our preliminary data showed that SLIT therapy in viro-immunological controlled HAART treated HIV positive patients was efficacious, safe and well tolerated.

Shrinking of silicon nanocrystals embedded in an amorphous silicon oxide matrix during rapid thermal annealing in a forming gas atmosphere.

We report the effect of hydrogen on the crystallization process of silicon nanocrystals embedded in a silicon oxide matrix. We show that hydrogen gas during annealing leads to a lower sub-band gap absorption, indicating passivation of defects created during annealing. Samples annealed in pure nitrogen show expected trends according to crystallization theory. Samples annealed in forming gas, however, deviate from this trend. Their crystallinity decreases for increased annealing time. Furthermore, we observe a decrease in the mean nanocrystal size and the size distribution broadens, indicating that hydrogen causes a size reduction of the silicon nanocrystals.

Circulating tumour cells as tumour biomarkers in melanoma: detection methods and clinical relevance.

Circulating tumour cells (CTCs) are cells of solid tumour origin detectable in the peripheral blood. Their occurrence is considered a prerequisite step for establishing distant metastases. Metastatic melanoma was the first malignancy in which CTCs were detected and numerous studies have been published on CTC detection in melanoma at various stages of disease. In spite of this, there is no general consensus as to the clinical utility of CTCs in melanoma, largely due to conflicting results from heterogeneous studies and discrepancies in methods of detection between studies. In this review, we examine the possible clinical significance of CTCs in cutaneous, mucosal and ocular melanoma, focusing on detection methods and prognostic value of CTC detection.

A case of anaphylaxis to Pollinex® Quattro MPL-4.

We described the first case reported in literature of anaphylactic shock after administration of pollen extract vaccine chemically modified (allergoid) adsorbed onto L-Tyrosine depot adjuvanted with monophosphoryl lipid A (Pollinex® Quattro MPL-4).

The presence of circulating tumor cells (CTCs) correlates with lymph node metastasis in nonresectable squamous cell carcinoma of the head and neck region (SCCHN).

BACKGROUND: The mechanisms regulating tumor cell dissemination in locally advanced squamous cell carcinoma of the head and neck region (SCCHN) are largely unresolved. We assessed the frequency of circulating tumor cells (CTCs), their association with clinicopathologic parameters and their kinetics during radiochemotherapy. PATIENTS AND METHODS: Peripheral blood samples from 42 patients with locally advanced SCCHN were included. CTCs were detected using flow cytometric analysis of CD45-epithelial cell adhesion molecule+cytokeratin+ cells and results were validated by nested RT-PCR analysis of circulating epidermal growth factor receptor transcripts. The association between the presence of CTCs and T stage, tumor volume, N stage and human papillomavirus status was evaluated. The influence of radiochemotherapy on CTC numbers was determined. RESULTS: CTCs were detected in 18 of 42 SCCHN patients (43%), with a mean ± standard deviation of 1.7 ± 0.9 CTCs per 3.75 ml blood. We observed no significant correlation between the presence of CTCs and T stage or tumor volume. However, a nodal stage of N2b or higher was associated with higher frequency of CTCs. Though concurrent radiochemotherapy reduced their frequency, CTCs persisted during treatment in 20% of cases. CONCLUSIONS: Detection of CTCs correlates with regional metastasis in inoperable SCCHN. Further follow-up is needed to evaluate the prognostic significance of CTC detection, in addition to clinical staging of lymph nodes, for regional or distant recurrence.

Expression of the stem cell marker nestin in peripheral blood of patients with melanoma.

BACKGROUND: There is continued interest in markers indicative of circulating melanoma cells. Nestin is a neuroepithelial intermediate filament protein that was found to be expressed in melanoma and in various cancer stem cells. OBJECTIVES: We investigated expression of nestin in peripheral blood of patients with melanoma. METHODS: We analysed nestin expression by flow cytometry and by quantitative reverse transcription-polymerase chain reaction both in tissues (n = 23) and in blood samples (n = 102) from patients with American Joint Committee on Cancer stage III-IV melanoma. Forty-six negative controls were also added. RESULTS: Flow cytometry did not reveal nestin-expressing cells in peripheral blood of healthy volunteers. In patients with melanoma, however, nestin protein was expressed in a proportion of melanoma cells enriched from peripheral blood by immunomagnetic sorting. In melanoma tissue samples a significant correlation was found between mRNAs coding for nestin and tyrosinase (P = 0.001) and melan-A (P = 0.002), whereas in blood a significant correlation was observed only for tyrosinase (P = 0.015), but not for melan-A (P = 0.53). Nestin expression was higher in stage IV patients compared with stage III/IV with no evidence of disease, in patients with high tumour burden, and was positively correlated to expression of tyrosinase and melan-A. CONCLUSIONS: Nestin was found to be an additional marker of interest for circulating melanoma cells. Prospective studies should investigate its potential added informative value in comparison with markers already in use for melanoma cell detection.

Immunological effects of omalizumab in chronic urticaria: a case report.

We describe a case of chronic idiopathic urticaria in which symptoms improved dramatically after treatment with omalizumab. This drug, which is approved for the treatment of asthma, has been studied in other allergic conditions and a number of reports have described its efficacy as an immunomodulator in chronic and physical urticaria. Immunopathologic mechanisms are poorly understood. In chronic autoimmune urticaria, it has been postulated that this monoclonal antibody against immunoglobulin (Ig) E might reduce FcepsilonRI expression on the surface of basophils, thus preventing IgG antibody-mediated crosslinking and the release of mast cell mediators. We analyzed activation and homing molecules of B cells and type 1 and type 2 cytokine production by T cells and document a new immunomodulator mechanism characterized by a reduction in B-cell activation and homing and in tumor necrosis factor-alpha and interleukin 4 production and an increase in interferon-gamma synthesis.

Three cases of delirium after "ecstasy" ingestion.

Three cases of delirium experienced by three young friends after recreational use of "ecstasy" are reported--a syndrome which, to the best of the authors' knowledge, has not been previously observed in MDMA abusers. Special attention is given to the etiological factors and clinical features of the adverse reaction.

Multicenter phase III randomized trial of polychemotherapy (CVD regimen) versus the same chemotherapy (CT) plus subcutaneous interleukin-2 and interferon-alpha2b in metastatic melanoma.

BACKGROUND: The addition of cytokines to chemotherapy (CT) has obtained encouraging but contradictory results in metastatic melanoma. In this phase III trial, we compared the effects of CT [cisplatin, vindesine and dacarbazine (CVD)] with those of concurrent biochemotherapy (bioCT) consisting of CVD plus interleukin-2 and interferon-alpha2b. PATIENTS AND METHODS: A total of 151 untreated metastatic melanoma patients were randomized, 75 on arm A (cisplatin 30 mg/m2 on days 1-3, vindesine 2.5 mg/m2 on day 1 and dacarbazine 250 mg/m2 on days 1-3), and 76 on arm B (same CVD scheme plus interferon-alpha2b on days 1-5 and interleukin-2 on days 1-5 and 8-15, both administered subcutaneously), either recycled every 3 weeks. Response was assessed every two cycles. RESULTS: Ten percent of the patients were alive at a median of 52 months from start of therapy. We observed a response rate (RR) of 21% on arm A versus 33% on arm B; three patients (4%) given bioCT had complete responses (CRs). Median time to progression (TTP) was identical; median overall survival (OS) time was 12 months on arm A and 11 months on arm B. CONCLUSIONS: BioCT is not better than CT alone; the trend in favor of the bioCT in terms of RR did not translate into better TTP or OS. Therefore, bioCT cannot be recommended as standard first-line therapy for metastatic melanoma.

Fast transient infrared studies in material science: development of a novel low dead-volume, high temperature DRIFTS cell.

A prototype DRIFTS flow reaction chamber was designed and developed in order to find analytical application in the study of heterogeneous catalysts operating at high temperatures under fast transient gas feed conditions. Minimisation of dead-volumes allows gas replacement in 8-10s at 10mLmin(-1) total flow. To overcome problems related to the reactivity of the cell walls under alternating oxidizing/reducing gases, the cell was built with Inconel 600trade mark, which was tested to be very inert even at high temperatures. The sample holder, which was developed to closely resemble a micro plug-flow reactor, poses some problems in terms of heat transfer to the outer body of the cell (limiting then the maximum reachable temperature) and of the correct measurement of the actual sample temperature. These problems were solved with a careful re-design of the upper part of the cell. The second prototype thus derived is able to reach temperatures up to 803K and allows gas replacement in less than 4s at 10mLmin(-1). The cell is inserted in a MCT-FT-IR, which allows to collect high quality spectra with a 1s time-resolution. The downstream flow can be analysed by a quadrupole mass spectrometer equipped with an enclosed source and by a commercial GC. The performances of this prototype cell are presented showing some tests carried out with ceria-zirconia (Ce(x)Zr(1-x)O(2)) catalysts for CO abatement under real operando conditions.

[Automatic voice recognition for radiologic reports. Experiment with a prototype in an operational environment]. / Il riconoscimento automatico della voce per la refertazione radiologica. Esperimento di un prototipo in ambiente operativo.

Large vocabulary speech recognition, its techniques and its software and hardware technology, are being developed, aimed at providing the office user with a tool that could significantly improve both quantity and quality of his work: the dictation machine, which allows memos and documents to be input using voice and a microphone instead of fingers and a keyboard. The IBM Rome Science Center, together with the IBM Research Division, has built a prototype recognizer that accepts sentences in natural language from a 20,000-word Italian vocabulary. The unit runs on a personal computer equipped with a special hardware capable of giving all the necessary computing power. The first laboratory experiments yielded very interesting results and pointed out such system characteristics to make its use possible in operational environments. To this purpose, the dictation of medical reports was considered as a suitable application. In cooperation with the 2nd Radiology Department of S. Maria della Misericordia Hospital (Udine, Italy), a system was experimented by radiology department doctors during their everyday work. The doctors were able to directly dictate their reports to the unit. The text appeared immediately on the screen, and eventual errors could be corrected either by voice or by using the keyboard. At the end of report dictation, the doctors could both print and archive the text. The report could also be forwarded to hospital information system, when the latter was available. Our results have been very encouraging: the system proved to be robust, simple to use, and accurate (over 95% average recognition rate). The experiment was precious for suggestions and comments, and its results are useful for system evolution towards improved system management and efficiency.

New-onset delusions and hallucinations in patients infected with HIV.

OBJECTIVE: To assess the relationship between HIV-associated psychotic symptoms (i.e., delusions, hallucinations) and demographic, psychopathological and medical variables by comparing patients with and without cerebral opportunistic infections or metabolic encephalopathy. DESIGN: Cross-sectional study. PATIENTS: 26 patients admitted to hospital with HIV and new-onset psychotic symptoms, defined according to DSM-III-R criteria. OUTCOME MEASURES: A semistructured psychiatric interview using the Psychopathology Assessment Scale (AMDP-4) of the Association for Methodology and Documentation in Psychiatry system. Comprehensive medical assessments, including laboratory tests and computed tomographic scans, were also performed. RESULTS: Patients with cerebral opportunistic infections or metabolic encephalopathy (i.e., "secondary" psychosis, n = 13) were more likely to show disorders of consciousness, disorders of orientation and disturbances of attention and memory than those with no evidence of HIV-related cerebral disease (i.e., "primary" psychosis, n = 13); 10 patients (77%) with cerebral opportunistic infections or metabolic encephalopathy and only 1 (8%) patient without (p < 0.001) were diagnosed with delirium. These associations were stronger for the "secondary" patients with no focal brain lesions than for those with lesions. CONCLUSIONS: These findings suggest that "organic" symptoms of psychosis in those infected with HIV are related to the systemic and cerebral complications of HIV infection rather than to the psychotic process itself.