High performance of multiplex fluorescence in situ hybridization to simultaneous detection of BCL2 and BCL6 rearrangements: useful application in the characterization of DLBCLs

Chromosomal rearrangements involving BCL2, BCL6 and MYC are commonly found in the most frequent B cell lymphomas, namely follicular lymphomas (FLs) and diffuse large B-cell lymphomas (DLBCLs). Particularly, BCL2-rearrangement represents a diagnostic hallmark in FLs, whereas MYC translocation can occur simultaneously with BCL2 and/or BCL6 rearrangements, defining a specific category of DLBCLs with a poorer prognosis. In this study, we aim to validate the diagnostic performance of multiplex BCL2/BCL6 FISH approach in formalin-fixed paraffin-embedded FLs and DBCLs and cytological samples of DLBCL comparing to the classic set of single break-apart probes. We collected a series of lymphomas, including 85 DLBCLs, 45 FLs and 36 other B-cell lymphoma histotypes and 16 cytological samples of DLBCLs. MYC, BCL2 and BCL6 rearrangements were previously assessed by a classic FISH test using single break-apart probes. All samples were analysed by a multiplex FISH assay. In the FL series, 38 cases showed BCL2-R; in the DLBCLs series, MYC-R was detected in 21 out of 85 DLBCL patients, BCL2-R in 10 out of 85 and BCL6-R in 33 out of 85. In the DLBCL cytological series, MYC-R was detected in 4 out of 16, BCL2-R in 4 out of 16 and BCL6-R in 1 out of 16. Notably, in FFPE, 13 double-hit lymphomas (DHLs) and 3 triple-hit lymphomas (THLs) were detected; in the cytological series, only 3 DHL cases were observed. The dual BCL2/BCL6 FISH probe test results were fully concordant with the results obtained using classic BCL2 and BCL6 single break apart. Particularly, multiplex FISH to simultaneously detect BCL2-R and BCL6-R on a single slide could find a wide application in the characterisation of double- and triple-hit DLBCLs.

Development of the Vacuum Arc Chute for a 110 kV Single-Break Vacuum Circuit Breaker

Requirements for high-voltage vacuum circuit breakers for networks with a grounded neutral, which are stemming from the prospects of using vacuum circuit breakers in networks with the solidly grounded neutral are formulated. The arch chute design for a 110 kV single-break vacuum circuit breaker that takes into account the occurrence of repeated breakdowns is developed. The shielding system equalizes the voltage over the chute casing, decreases the electric field intensity at the “triple point”, protects the casing dielectric from becoming dusted with metallic contact parts erosion products, prevents the occurrence of cascade breakdown between the contact parts and central shield, and eliminates a possible decrease of the chute electric strength as it gradually works out its switching life. It is shown that the chute design includes measures to prevent the possibility of repeated breakdowns during disconnection up to class С1 according to GOST 52565-2006. It has been proven that for vacuum arc chutes for class С2 (for switching a capacitive load) it is necessary to develop a chute with two breaks between the contact parts or to connect two vacuum chutes in series. The shielding system adopted in the chute rules out the possibility of ceramic casing to become metallized, which decreases the chute electric strength as it gradually works out its switching life, and the vacuum arc transfer to beyond the inter-contact gap boundaries onto the central shield, the burnout of which results in that the vacuum arc chute loses its tightness and fully loses its functional properties.

Experimental Investigation of the Prestrike Characteristics of a Double-Break Vacuum Circuit Breaker under DC Voltages

The prestrike phenomenon in vacuum circuit breakers (VCBs) is interesting but complicated. Previous studies mainly focus on the prestrike phenomenon in single-break VCBs. However, experimental work on prestrike characteristics of double-break VCBs cannot be found in literature. This paper aims to experimentally determine the probabilistic characteristics of prestrike gaps in a double-break VCB consisting of two commercial vacuum interrupters (VIs) in series under direct current (DC) voltages. As a benchmark, single-break prestrike gaps were measured by short-circuiting one of the VIs in a double break. The experimental results show that the 50% prestrike gap d50 of each VI in a double break, which is calculated with the complementary Weibull distribution, was significantly reduced by 25% to 72.7% compared with that in a single break. Due to the voltage-sharing effect in the double-break VCB, scatters in prestrike gaps of each VI in a double break was smaller than that in a single break. However, without the sharing-voltage effect, d50 of the low-voltage side in the double break was 65% higher than that of the same VI in the single break, which could be caused by the asynchronous property of mechanical actuators, the difference of the inherent prestrike characteristics of each VI and the unequal voltage-sharing ratio of VIs.

Multiplex fluorescence in situ hybridisation to detect anaplastic lymphoma kinase and ROS proto-oncogene 1 receptor tyrosine kinase rearrangements in lung cancer cytological samples

AimsSeveral predictive biomarkers of response to specific inhibitors have become mandatory for the therapeutic choice in non-small-cell lung cancer (NSCLC). In most lung cancer patients, the biological materials available to morphological and molecular diagnosis are exclusively cytological samples and minimum tumour wastage is necessary. Multiplex fluorescence in situ hybridisation (mFISH) to detect simultaneously ALK-rearrangement and ROS1-rearrangement on a single slide could be useful in clinical practice to save cytological samples for further molecular analysis. In this study, we aim to validate diagnostic performance of multiplex ALK/ROS1 fluorescence in situ hybridisation (FISH) approach in lung adenocarcinoma cytological series compared with classic single break apart probes.MethodsWe collected a series of 61 lung adenocarcinoma cytological specimens enriched in tumours harbouring ALK-rearrangement and ROS1-rearrangement. ALK and ROS1 status were previously assessed by classic FISH test using single break apart probes and immunohistochemistry. Study population was composed of 6 ALK-positive, 2 ROS1-positive and 53 ALK/ROS1-wild type. All specimens were analysed by multiplex FISH assay using FlexISH ALK/ROS1 DistinguISH Probe Zytovision.ResultsThe dual ALK/ROS1 FISH probe test results were fully concordant with the results of previous single ALK and ROS1 FISH tests on two different slides. 6 ALK-positive and 2 ROS1-positive were confirmed through multiplex FISH test, without false-positive and false-negative results. Multiplex ALK/ROS1 FISH test results agreed with immunohistochemistry assay staining results.ConclusionMultiplex ALK/ROS1 FISH probe test is a useful tool to detect simultaneously ALK-rearrangement and ROS1-rearrangement on a single slide in cytological specimens with a small amount of biomaterial.