Melanoma NOS1 expression promotes dysfunctional IFN signaling.

In multiple forms of cancer, constitutive activation of type I IFN signaling is a critical consequence of immune surveillance against cancer; however, PBMCs isolated from cancer patients exhibit depressed STAT1 phosphorylation in response to IFN-α, suggesting IFN signaling dysfunction. Here, we demonstrated in a coculture system that melanoma cells differentially impairs the IFN-α response in PBMCs and that the inhibitory potential of a particular melanoma cell correlates with NOS1 expression. Comparison of gene transcription and array comparative genomic hybridization (aCGH) between melanoma cells from different patients indicated that suppression of IFN-α signaling correlates with an amplification of the NOS1 locus within segment 12q22-24. Evaluation of NOS1 levels in melanomas and IFN responsiveness of purified PBMCs from patients indicated a negative correlation between NOS1 expression in melanomas and the responsiveness of PBMCs to IFN-α. Furthermore, in an explorative study, NOS1 expression in melanoma metastases was negatively associated with patient response to adoptive T cell therapy. This study provides a link between cancer cell phenotype and IFN signal dysfunction in circulating immune cells

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]

Discriminating between “drizzle or rain” and sea salt aerosols in Cloudnet for measurements over the Barbados Cloud Observatory

The highly sensitive Ka-band cloud radar at the Barbados Cloud Observatory (BCO) frequently reveals radar reflectivity signals below −50 dBZ within the convective sub-cloud layer. These so-called haze echoes are signals from hygroscopically grown sea salt aerosols. Within the Cloudnet target classification scheme, haze echoes are generally misclassified as precipitation (target class: “drizzle or rain”). We present a technique to discriminate between “drizzle or rain” and sea salt aerosols in Cloudnet that is applicable to marine Cloudnet sites. The method is based on deriving heuristic probability functions utilizing a combination of cloud radar reflectivity factor, radar mean Doppler velocity, and the ceilometer attenuated backscatter coefficient. The method is crucial for investigating the occurrence of precipitation and significantly improves the Cloudnet target classification scheme for measurements at the BCO. The results are validated against the amount of precipitation detected by the Virga-Sniffer tool. We analyze data for measurements at BCO covering 2 years (July 2021–July 2023). A first-ever statistical analysis of the Cloudnet target classification product including the new “haze echo” target over 2 years at the BCO is presented. In the atmospheric column above the BCO, “drizzle or rain” is on average more frequent during the dry season compared to the wet season due to the higher occurrence of warm clouds contributing to the amount of precipitation. Haze echoes are identified about 4 times more often during the dry season compared to the wet season. The frequency of occurrence of “drizzle or rain” in Cloudnet caused by misclassified haze echoes is overestimated by up to 16 %. Supported by the Cloudnet statistics and the results obtained from the Virga-Sniffer tool, 48 % of detected warm clouds in the dry and wet season precipitate. The proportion of precipitation evaporating fully before reaching the ground (virga) is higher during the dry season.</p

Impact of wildfire smoke on Arctic cirrus formation – Part 1: Analysis of MOSAiC 2019–2020 observations

The potential impact of wildfire smoke on Arctic cirrus formation is discussed based on lidar and radar observations during the winter half year of the 1-year MOSAiC (Multidisciplinary drifting Observatory for the Study of Arctic Climate) expedition. Aerosol and ice cloud observations were performed aboard the icebreaker Polarstern at latitudes > 85° N. Aged Siberian wildfire smoke polluted the tropopause region over the central Arctic during the entire winter half year of 2019-2020. The smoke particle surface area concentration at the tropopause was of the order of 5-15 μm2cm-3 and indicated considerably enhanced levels of aerosol pollution for more than 6 months. Numerous cirrus systems with cloud-top temperatures between -60 and -75 °C developed in the polluted upper troposphere. We analyzed all MOSAiC winter cirrus layers with respect to their geometrical and optical properties and a subgroup of 20 cirrus events with respect to their ice water content (IWC) and ice crystal number concentration (ICNC). In individual ice fallstreaks that are connected to individual ice nucleation events, ICNCs typically ranged from 1 to 10 crystals L-1 but were frequently also as high as 20-50 L-1; however, observations > 100 L-1 were rare. Three observational facts corroborate our hypothesis that smoke significantly influenced Arctic cirrus formation: (1) the occurrence of a long-lasting, persistent smoke pollution layer in the upper troposphere so that favorable conditions for heterogeneous ice nucleation on smoke particles were always given and, at the same time, homogeneous freezing of background aerosol was probably widely suppressed; (2) the high smoke particle surface area concentrations, which were high enough to significantly trigger ice nucleation on smoke particles (as shown in Part 2, the companion paper to this article; ); and (3) the frequently found maximum cirrus ice saturation ratios of 1.3-1.5, which point to the dominance of heterogeneous ice nucleation processes, initiated by inefficient ice-nucleating particles (INPs), as expected when aged smoke particles (i.e., organic aerosol particles) serve as INPs. The studies are continued in the simulation portion of this work (Part 2; Ansmann et al., 2025)

Identification of gene polymorphisms of human DNA topoisomerase I in the National Cancer Institute panel of human tumour cell lines

Topoisomerase 1 (Top1), a nuclear enzyme involved in DNA relaxation, is the target of several anticancer drugs. TOP1 mutations occur in camptothecin-resistant tumour cell lines. We explored, in the NCI panel of 60 human tumour cell lines, whether polymorphic variations in the TOP1 gene could explain differences in drug sensitivity. The 21 exons of the gene were fully studied as well as five intronic domains that had previously been shown to harbour single nucleotide polymorphisms (SNPs) or mutations. PCR products covering the whole exonic sequences or the relevant intronic domains were subjected to denaturing high-performance liquid chromatography. Nucleotide variations were then determined by sequencing. Discrimination between intronic common and variant homozygous samples was performed using a restriction fragment length polymorphism technique. Only one exonic mutation was detected, at the heterozygous state; it occurs in exon 19 of a colon cancer cell line (HCT-15) and consists of a G>A transition at position 75, resulting in a Met675Ile change. The intronic sequences studied harboured the SNPs expected with allelic frequencies between 20 and 40%. Three major haplotypes, generating 92% of the 10 genotypes encountered, were defined as containing none of the intronic SNPs, or three of them, or all of them. No significant relationship was evidenced between Top1 expression and the TOP1 polymorphisms studied. However, when comparing the cytotoxicity of 138 drugs as a function of the genotypes, several drug groups, namely Top1 inhibitors, antifolates and taxanes, had significantly different IC50s as a function of the distribution of the intronic SNPs of the TOP1 gene

Dielectrophoresis has Broad Applicability to Marker-Free Isolation of Tumor Cells from Blood by Microfluidic Systems

The number of circulating tumor cells (CTCs) found in blood is known to be a prognostic marker for recurrence of primary tumors, however, most current methods for isolating CTCs rely on cell surface markers that are not universally expressed by CTCs. Dielectrophoresis (DEP) can discriminate and manipulate cancer cells in microfluidic systems and has been proposed as a molecular marker-independent approach for isolating CTCs from blood. To investigate the potential applicability of DEP to different cancer types, the dielectric and density properties of the NCI-60 panel of tumor cell types have been measured by dielectrophoretic field-flow fractionation (DEP-FFF) and compared with like properties of the subpopulations of normal peripheral blood cells. We show that all of the NCI-60 cell types, regardless of tissue of origin, exhibit dielectric properties that facilitate their isolation from blood by DEP. Cell types derived from solid tumors that grew in adherent cultures exhibited dielectric properties that were strikingly different from those of peripheral blood cell subpopulations while leukemia-derived lines that grew in non-adherent cultures exhibited dielectric properties that were closer to those of peripheral blood cell types. Our results suggest that DEP methods have wide applicability for the surface-marker independent isolation of viable CTCs from blood as well as for the concentration of leukemia cells from blood. (C) 2013 American Institute of Physics. [http://dx.doi.org/10.1063/1.4774307]Cancer Prevention and Research Institute of Texas (CPRIT) RP100934Kleberg Center for Molecular MarkersEntertainment Industry Foundation SU2C-AACR-DT0209NCI CA016672Biomedical Engineerin

Active centromere and chromosome identification in fixed cell lines

BACKGROUND: The centromere plays a crucial role in ensuring the fidelity of chromosome segregation during cell divisions. However, in cancer and constitutional disorders, the presence of more than one active centromere on a chromosome may be a contributing factor to chromosome instability and could also have predictive value in disease progression, making the detection of properly functioning centromeres important. Thus far, antibodies that are widely used for functional centromere detection mainly work on freshly harvested cells whereas most cytogenetic samples are stored long-term in methanol-acetic acid fixative. Hence, we aimed to identify antibodies that would recognise active centromere antigens on methanol-acetic acid fixed cells. RESULTS: A panel of active centromere protein antibodies was tested and we found that a rabbit monoclonal antibody against human CENP-C recognises the active centromeres of cells fixed in methanol-acetic acid. We then tested and compared combinations of established methods namely centromere fluorescence in situ hybridisation (cenFISH), centromere protein immunofluorescence (CENP-IF) and multicolour FISH (mFISH), and showed the usefulness of CENP-IF together with cenFISH followed by mFISH (CENP-IF-cenFISH-mFISH) with the aforementioned anti-CENP-C antibody. We further demonstrated the utility of our method in two cancer cell lines with high proportion of centromere defects namely neocentromere and functional dicentric. CONCLUSIONS: We propose the incorporation of the CENP-IF-cenFISH-mFISH method using a commercially available rabbit monoclonal anti-CENP-C into established methods such as dicentric chromosome assay (DCA), prenatal karyotype screening in addition to constitutional and cancer karyotyping. This method will provide a more accurate assessment of centromere abnormality status in chromosome instability disorders