Disseminated cancer cells detected by immunocytology in lymph nodes of NSCLC patients are highly prognostic and undergo parallel molecular evolution.

In melanoma, immunocytology (IC) after sentinel lymph node disaggregation not only enables better quantification of disseminated cancer cells (DCCs) than routine histopathology (HP) but also provides a unique opportunity to detect, isolate, and analyse these earliest harbingers of metachronous metastasis. Here, we explored lymph node IC in non-small cell lung cancer (NSCLC). For 122 NSCLC patients, 220 lymph nodes (LNs) were split in half and prepared for IC and HP. When both methods were compared, IC identified 22% positive patients as opposed to 4.5% by HP, revealing a much higher sensitivity of IC (p < 0.001). Assessment of all available 2,952 LNs of the same patients by HP uncovered additional patients escaping detection of lymphatic tumour spread by IC alone, consistent with the concept of skip metastasis. A combined lymph node status of IC and complete HP on a larger cohort of patients outperformed all risk factors in multivariable analysis for prognosis (p < 0.001; RR = 2.290; CI 1.407-3.728). Moreover, isolation of DCCs and single-cell molecular characterization revealed that (1) LN-DCCs differ from primary tumours in terms of copy number alterations and selected mutations and (2) critical alterations are acquired during colony formation within LNs. We conclude that LN-IC in NSCLC patients when combined with HP improves diagnostic precision, has the potential to reduce total workload, and facilitates molecular characterization of lymphatically spread cancer cells, which may become key for the selection and development of novel systemic therapies. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Wind Tunnel Performance Tests of the Propellers with Different Pitch for the Electric Propulsion System.

The geometry of a propeller is closely related to its aerodynamic performance. One of the geometric parameters of a propeller is pitch. This parameter determines the distance by which the propeller moves forward during one revolution. The challenge is to select a propeller geometry for electric propulsion in order to achieve the best possible performance. This paper presents the experimental results of the aerodynamic performance of the set of propellers with different pitch values. The tests were performed in a closed-circuit subsonic wind tunnel using a six-component force balance. The analyzed propellers were 12-inch diameter twin-blade propellers that were driven by a BLDC (brushless direct current) electric motor. The tests were performed under forced airflow conditions. The thrust and torque produced by the propeller were measured using a strain gauge. The analysis was performed for different values of the advance ratio which is the ratio of freestream fluid speed to propeller tip speed. Additionally, a set of electrical parameters was recorded using the created measurement system. The propeller performance was evaluated by a dimensional analysis. This method enables calculation of dimensionless coefficients which are useful for comparing performance data for propellers.

Measurements of Aerodynamic Interference of a Hybrid Aircraft with Multirotor Propulsion.

This article deals with the phenomenon of aerodynamic interference occurring in the innovative hybrid system of multirotor aircraft propulsion. The approach to aerodynamics requires a determination of the impact of active sources of lift and thrust upon the aircraft aerodynamic characteristics. The hybrid propulsion unit, composed of a conventional multirotor source of thrust as well as lift in the form of the main rotor and a pusher, was equipped with an additional propeller drive unit. The tests were conducted in a continuous-flow low speed wind tunnel with an open measuring space, 1.5 m in diameter and 2.0 m long. Force testing made it possible to develop aerodynamic characteristics as well as defining aerodynamic characteristics and defining the field of speed for the considered design configurations. Our investigations enabled us to analyze the results in terms of a mutual impact of particular components of the research object and the area of impact of active elements present in a common flow.

Measurement of the Flow Field Generated by Multicopter Propellers.

This paper presents the results of research on the airflow around a multirotor aircraft. The research consisted of the analysis of the velocity field using particle image velocimetry. Based on the tests carried out in a wind tunnel, the distribution of the velocity and its components in the vertical plane passing through the propeller axis were determined for several values of the angle of attack of the tested object for two values of airflow velocity inside the tunnel, i.e., vwt = 0 m/s and vwt = 12.5 m/s. Determining the velocity value as a function of the coordinates of the adopted reference system allowed for defining the range of impact of the horizontal propellers and the fuselage of the research object itself. The tests allowed for quantitative and qualitative analyses of the airflow through the horizontal rotor. Particular attention was paid to the impact of the airflow and the angle of attack on the obtained velocity field distributions.

HER2 FISH results in breast cancers with increased CEN17 signals using alternative chromosome 17 probes - reclassifying cases in the equivocal category.

AIMS: HER2 testing of invasive breast cancer by in-situ hybridization guides therapy decisions. Probing HER2 and centromere of chromosome 17 (cen17) simultaneously is supposed to reveal both a potential HER2 gene amplification and polysomy 17. However, a considerable number of breast cancer patients with quasi polysomy 17 are considered 'equivocal', which is diagnostically meaningless. Moreover, patients with equivocal/false polysomic tumours are prevented from a potentially beneficial anti-HER2 treatment. Here we evaluated the RAI1, D17S122 and TP53 hybridization markers to indicate true polysomy reliably and to reclassify equivocal samples accurately as HER2-positive. METHODS AND RESULTS: Samples with (n = 82) and without (n = 52) increased cen17 copy numbers and 78 evidently HER2-amplified specimens were analysed using dual and triple marker hybridization probes. Selected putative polysomic samples were subjected to array-based comparative genomic hybridization (aCGH). We found that 37.8% samples with putative polysomy 17 did not show any gain in RAI1, D17S122 or TP53. Accordingly, aCGH revealed evidence for the presence of HER2/cen17 co-amplification rather than for true polysomy in those cases. Reflex testing using alternate 17p markers reclassified up to 56.3% equivocal cases as HER2-positive and the combined assessment of a 17p and 17q locus allows the differentiation of true versus false polysomy. CONCLUSIONS: An increased cen17 copy number does not necessarily reflect polysomy, and reflex testing facilitates the reclassification of 'equivocals'. Nevertheless, the prognostic and predictive value of a HER2/cen17 co-amplification versus HER2 gene amplification alone must still be evaluated prospectively.

Quantitative measurement of melanoma spread in sentinel lymph nodes and survival.

BACKGROUND: Sentinel lymph node spread is a crucial factor in melanoma outcome. We aimed to define the impact of minimal cancer spread and of increasing numbers of disseminated cancer cells on melanoma-specific survival. METHODS AND FINDINGS: We analyzed 1,834 sentinel nodes from 1,027 patients with ultrasound node-negative melanoma who underwent sentinel node biopsy between February 8, 2000, and June 19, 2008, by histopathology including immunohistochemistry and quantitative immunocytology. For immunocytology we recorded the number of disseminated cancer cells (DCCs) per million lymph node cells (DCC density [DCCD]) after disaggregation and immunostaining for the melanocytic marker gp100. None of the control lymph nodes from non-melanoma patients (n = 52) harbored gp100-positive cells. We analyzed gp100-positive cells from melanoma patients by comparative genomic hybridization and found, in 45 of 46 patients tested, gp100-positive cells displaying genomic alterations. At a median follow-up of 49 mo (range 3-123 mo), 138 patients (13.4%) had died from melanoma. Increased DCCD was associated with increased risk for death due to melanoma (univariable analysis; p<0.001; hazard ratio 1.81, 95% CI 1.61-2.01, for a 10-fold increase in DCCD + 1). Even patients with a positive DCCD ≤3 had an increased risk of dying from melanoma compared to patients with DCCD = 0 (p = 0.04; hazard ratio 1.63, 95% CI 1.02-2.58). Upon multivariable testing DCCD was a stronger predictor of death than histopathology. The final model included thickness, DCCD, and ulceration (all p<0.001) as the most relevant prognostic factors, was internally validated by bootstrapping, and provided superior survival prediction compared to the current American Joint Committee on Cancer staging categories. CONCLUSIONS: Cancer cell dissemination to the sentinel node is a quantitative risk factor for melanoma death. A model based on the combined quantitative effects of DCCD, tumor thickness, and ulceration predicted outcome best, particularly at longer follow-up. If these results are validated in an independent study, establishing quantitative immunocytology in histopathological laboratories may be useful clinically.

Array-Based Comparative Genomic Hybridization for the Detection of Copy Number Alterations in Single Cells.

Comprehensive genome-wide analyses of single cells represent an important tool for clinical applications, such as pre-implantation diagnostic and prenatal diagnosis, as well as for cancer research purpose. For the latter, studies of tumor heterogeneity, circulating tumor cells (CTCs), and disseminated cancer cells (DCCs) require the analysis of single-cell genomes. Here we describe a reliable and robust array-based comparative genomic hybridization (aCGH) protocol based on Ampli 1™ whole genome amplification that allows the detection of copy number alterations (CNAs) in single cancer cells as small as 100 kb.

Ceramic-Based Piezoelectric Material for Energy Harvesting Using Hybrid Excitation.

This paper analyzes the energy efficiency of a Micro Fiber Composite (MFC) piezoelectric system. It is based on a smart Lead Zirconate Titanate material that consists of a monolithic PZT (piezoelectric ceramic) wafer, which is a ceramic-based piezoelectric material. An experimental test rig consisting of a wind tunnel and a developed measurement system was used to conduct the experiment. The developed test rig allowed changing the air velocity around the tested bluff body and the frequency of forced vibrations as well as recording the output voltage signal and linear acceleration of the tested object. The mechanical vibrations and the air flow were used to find the optimal performance of the piezoelectric energy harvesting system. The performance of the proposed piezoelectric wind energy harvester was tested for the same design, but of different masses. The geometry of the hybrid bluff body is a combination of cuboid and cylindrical shapes. The results of testing five bluff bodies for a range of wind tunnel air flow velocities from 4 to 15 m/s with additional vibration excitation frequencies from 0 to 10 Hz are presented. The conducted tests revealed the areas of the highest voltage output under specific excitation conditions that enable supplying low-power sensors with harvested energy.

Retrospective cell lineage reconstruction in humans by using short tandem repeats.

Cell lineage analysis aims to uncover the developmental history of an organism back to its cell of origin. Recently, novel in vivo methods utilizing genome editing enabled important insights into the cell lineages of animals. In contrast, human cell lineage remains restricted to retrospective approaches, which still lack resolution and cost-efficient solutions. Here, we demonstrate a scalable platform based on short tandem repeats targeted by duplex molecular inversion probes. With this human cell lineage tracing method, we accurately reproduced a known lineage of DU145 cells and reconstructed lineages of healthy and metastatic single cells from a melanoma patient who matched the anatomical reference while adding further refinements. This platform allowed us to faithfully recapitulate lineages of developmental tissue formation in healthy cells. In summary, our lineage discovery platform can profile informative somatic mutations efficiently and provides solid lineage reconstructions even in challenging low-mutation-rate healthy single cells.

Interleukin-6 trans-signaling is a candidate mechanism to drive progression of human DCCs during clinical latency.

Although thousands of breast cancer cells disseminate and home to bone marrow until primary surgery, usually less than a handful will succeed in establishing manifest metastases months to years later. To identify signals that support survival or outgrowth in patients, we profile rare bone marrow-derived disseminated cancer cells (DCCs) long before manifestation of metastasis and identify IL6/PI3K-signaling as candidate pathway for DCC activation. Surprisingly, and similar to mammary epithelial cells, DCCs lack membranous IL6 receptor expression and mechanistic dissection reveals IL6 trans-signaling to regulate a stem-like state of mammary epithelial cells via gp130. Responsiveness to IL6 trans-signals is found to be niche-dependent as bone marrow stromal and endosteal cells down-regulate gp130 in premalignant mammary epithelial cells as opposed to vascular niche cells. PIK3CA activation renders cells independent from IL6 trans-signaling. Consistent with a bottleneck function of microenvironmental DCC control, we find PIK3CA mutations highly associated with late-stage metastatic cells while being extremely rare in early DCCs. Our data suggest that the initial steps of metastasis formation are often not cancer cell-autonomous, but also depend on microenvironmental signals.

Targeted transcript quantification in single disseminated cancer cells after whole transcriptome amplification.

Gene expression analysis of rare or heterogeneous cell populations such as disseminated cancer cells (DCCs) requires a sensitive method allowing reliable analysis of single cells. Therefore, we developed and explored the feasibility of a quantitative PCR (qPCR) assay to analyze single-cell cDNA pre-amplified using a previously established whole transcriptome amplification (WTA) protocol. We carefully selected and optimized multiple steps of the protocol, e.g. re-amplification of WTA products, quantification of amplified cDNA yields and final qPCR quantification, to identify the most reliable and accurate workflow for quantitation of gene expression of the ERBB2 gene in DCCs. We found that absolute quantification outperforms relative quantification. We then validated the performance of our method on single cells of established breast cancer cell lines displaying distinct levels of HER2 protein. The different protein levels were faithfully reflected by transcript expression across the tested cell lines thereby proving the accuracy of our approach. Finally, we applied our method to breast cancer DCCs of a patient undergoing anti-HER2-directed therapy. Here, we were able to measure ERBB2 expression levels in all HER2-protein-positive DCCs. In summary, we developed a reliable single-cell qPCR assay applicable to measure distinct levels of ERBB2 in DCCs.

In Vivo Detection of Circulating Tumor Cells in High-Risk Non-Metastatic Prostate Cancer Patients Undergoing Radiotherapy.

High-risk non-metastatic prostate cancer (PCa) has the potential to progress into lethal disease. Treatment options are manifold but, given a lack of surrogate biomarkers, it remains unclear which treatment offers the best results. Several studies have reported circulating tumor cells (CTCs) to be a prognostic biomarker in metastatic PCa. However, few reports on CTCs in high-risk non-metastatic PCa are available. Herein, we evaluated CTC detection in high-risk non-metastatic PCa patients using the in vivo CellCollector CANCER01 (DC01) and CellSearch system. CTC counts were analyzed and compared before and after radiotherapy (two sampling time points) in 51 high-risk non-metastatic PCa patients and were further compared according to isolation technique; further, CTC counts were correlated to clinical features. Use of DC01 resulted in a significantly higher percentage of CTC-positive samples compared to CellSearch (33.7% vs. 18.6%; p = 0.024) and yielded significantly higher CTC numbers (range: 0-15 vs. 0-5; p = 0.006). Matched pair analysis of samples between two sampling time points showed no difference in CTC counts determined by both techniques. CTC counts were not correlated with clinicopathological features. In vivo enrichment using DC01 has the potential to detect CTC at a higher efficiency compared to CellSearch, suggesting that CTC is a suitable biomarker in high-risk non-metastatic PCa.

Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization.

Rare target cells can be isolated from a high background of non-target cells using antibodies specific for surface proteins of target cells. A recently developed method uses a medical wire functionalized with anti-epithelial cell adhesion molecule (EpCAM) antibodies for in vivo isolation of circulating tumor cells (CTCs)1. A patient-matched cohort in non-metastatic prostate cancer showed that the in vivo isolation technique resulted in a higher percentage of patients positive for CTCs as well as higher CTC counts as compared to the current gold standard in CTC enumeration. As cells cannot be recovered from current medical devices, a new functionalized wire (referred to as Device) was manufactured allowing capture and subsequent detachment of cells by enzymatic treatment. Cells are allowed to attach to the Device, visualized on a microscope and detached using enzymatic treatment. Recovered cells are cytocentrifuged onto membrane-coated slides and harvested individually by means of laser microdissection or micromanipulation. Single-cell samples are then subjected to single-cell whole genome amplification allowing multiple downstream analysis including screening and target-specific approaches. The procedure of isolation and recovery yields high quality DNA from single cells and does not impair subsequent whole genome amplification (WGA). A single cell's amplified DNA can be forwarded to screening and/or targeted analysis such as array comparative genome hybridization (array-CGH) or sequencing. The device allows ex vivo isolation from artificial rare cell samples (i.e. 500 target cells spiked into 5 mL of peripheral blood). Whereas detachment rates of cells are acceptable (50 - 90%), the recovery rate of detached cells onto slides spans a wide range dependent on the cell line used (<10 - >50%) and needs some further attention. This device is not cleared for the use in patients.

Genetic alterations driving metastatic colony formation are acquired outside of the primary tumour in melanoma.

Mouse models indicate that metastatic dissemination occurs extremely early; however, the timing in human cancers is unknown. We therefore determined the time point of metastatic seeding relative to tumour thickness and genomic alterations in melanoma. Here, we find that lymphatic dissemination occurs shortly after dermal invasion of the primary lesion at a median thickness of ~0.5 mm and that typical driver changes, including BRAF mutation and gained or lost regions comprising genes like MET or CDKNA2, are acquired within the lymph node at the time of colony formation. These changes define a colonisation signature that was linked to xenograft formation in immunodeficient mice and death from melanoma. Thus, melanoma cells leave primary tumours early and evolve at different sites in parallel. We propose a model of metastatic melanoma dormancy, evolution and colonisation that will inform direct monitoring of adjuvant therapy targets.

Catch and Release: rare cell analysis from a functionalised medical wire.

Enumeration and especially molecular characterization of circulating tumour cells (CTCs) holds great promise for cancer management. We tested a modified type of an in vivo enrichment device (Catch&Release) for its ability to bind and detach cancer cells for the purpose of single-cell molecular downstream analysis in vitro. The evaluation showed that single-cell analysis using array comparative genome hybridization (array-CGH) and next generation sequencing (NGS) is feasible. We found array-CGH to be less noisy when whole genome amplification (WGA) was performed with Ampli1 as compared to GenomePlex (DLRS values 0.65 vs. 1.39). Moreover, Ampli1-processed cells allowed detection of smaller aberrations (median 14.0 vs. 49.9 Mb). Single-cell NGS data obtained from Ampli1-processed samples showed the expected non-synonymous mutations (deletion/SNP) according to bulk DNA. We conclude that clinical application of this refined in vivo enrichment device allows CTC enumeration and characterization, thus, representing a promising tool for personalized medicine.

Deterministic Whole-Genome Amplification of Single Cells.

This chapter describes a single-cell whole genome amplification method (WGA) that has been originally published under the name "Single Cell Comparative Genomic Hybridization (SCOMP)" (Klein et al., Proc Natl Acad Sci U S A 96(8):4494-4499, 1999). The method has recently become available commercially under the name "Ampli1(™) WGA Kit." It is a PCR-based technique for whole genome amplification (WGA) allowing comprehensive and quite uniform amplification of DNA from low quantities of input DNA material, in particular single cells. The method is based on a ligation-mediated adaptor linker PCR approach. In contrast to other PCR-based WGA approaches, both the primer design and mechanism underlying the fragmentation of genome are nonrandom, enabling high priming efficiency and deterministic fragmentation of template DNA. This is particularly important for the design of (diagnostic) assays targeting specific loci. Here, we describe the WGA protocol for amplification of single-cell genomes designed to provide high-quality material in quantity sufficient for a number of locus-specific and genome-wide downstream assays [e.g., targeted Sanger sequencing, restriction fragment length polymorphism (RFLP), quantitative PCR (qPCR), and array comparative genomic hybridization (CGH)].

Principles of Whole-Genome Amplification.

Modern molecular biology relies on large amounts of high-quality genomic DNA. However, in a number of clinical or biological applications this requirement cannot be met, as starting material is either limited (e.g., preimplantation genetic diagnosis (PGD) or analysis of minimal residual cancer) or of insufficient quality (e.g., formalin-fixed paraffin-embedded tissue samples or forensics). As a consequence, in order to obtain sufficient amounts of material to analyze these demanding samples by state-of-the-art modern molecular assays, genomic DNA has to be amplified. This chapter summarizes available technologies for whole-genome amplification (WGA), bridging the last 25 years from the first developments to currently applied methods. We will especially elaborate on research application, as well as inherent advantages and limitations of various WGA technologies.

Laser Microdissection of FFPE Tissue Areas and Subsequent Whole Genome Amplification by Ampli1™.

Laser microdissection (LMD) and whole genome amplification (WGA) are valuable tools to isolate, purify, and genetically analyze cancer cells from tissue sections. In this chapter, we describe a workflow for microdissecting small regions of interest from cancer tissue, i.e. formalin-fixed paraffin-embedded (FFPE) and cryo-conserved specimens, and subsequent whole genome amplification by a deterministic WGA approach (Ampli1™ WGA).

Time and frequency analysis of beat-to-beat R-T interval variability in patients with ischaemic left ventricular dysfunction providing evidence for non-neural control of ventricular repolarisation.

BACKGROUND: Determinants of temporal lability in ventricular repolarisation are not fully recognised. We aimed to analyse the sources of RT variability by comparing normal subjects and patients after myocardial infarction (MI) with either depressed or preserved left ventricular (LV) function. METHODS: One hundred and nine patients (27 women, 82 men, aged 51 +/- 9 years) were divided into three groups: 24 patients (pts) with an uncomplicated angiographically proven coronary heart disease (CHD-group), 59 post-MI pts with preserved LV function (LVEF > 40%, PMI-N-group) and 26 post-MI pts with depressed LV function (LVEF < 40%, PMI-L-group). An ECG signal of low-noise 512 heartbeats was recorded using a computer-assisted amplifier (16 bit, 2 kHz). The onset and offset of the R-wave and T-wave were determined automatically. The magnitude of R-R and R-T variability was measured as the standard deviation of all intervals (SD-RR and SD-RT, ms, respectively). Their relationship was quantified by the correlation coefficient r(RT/RR). Power spectral density of RR or RT variability was estimated with the FFT (Welch's averaged periodogram, Hanning window) and frequency relation was quantified using a squared coherence spectrum (SCS). For all spectral and cross-spectral measurements two frequency ranges were considered: high (0.15-0.50 Hz, HF) and low (0.04-0.15 Hz, LF). Spectral power and SCS of RR and RT variability for both ranges (HF(RR), LF(RR), HF(RT), LF(RT), SCS(HF), SCS(LF)), and the ratios LF/HF(RR) and LF/HF(RT) were drawn for comparisons. The central frequency of HF(RR) was considered as the frequency of respiration (f(resp), Hz). RESULTS: In the PMI-L group the SD-RT was significantly greater compared to the remaining groups and accounted for almost 10% of the SDRR. Also, the coefficient r(RT/RR) was weakest in this group. The spectral indices of RR variability were similar in all groups, while the greatest value of the HFRT was observed in the PMI-L group. The SCS(LF) was insignificant in this group, contrary to the CHD and PMI-N groups. Additionally, there were significant negative relationships between f(resp) and spectral indices of RT variability in PMI-patients with depressed LV function. CONCLUSION: A greater beat-to-beat variation in RT interval duration along with increased power of its HF component indicates an important role of respiration in ventricular repolarisation control, while reduced time- and frequency RT-RR relationships seem to relate to an impaired process of ventricular duration adaptation.

Age-adjustment of HRV measures and its prognostic value for risk assessment in patients late after myocardial infarction.

THE AIMS OF OUR STUDY WERE: (1) to establish the normal limit of the heart rate variability (HRV) indices in a healthy population and in its four age-related subgroups, including a new HRV index, HRV fraction; and (2) to analyse the frequency and predictive value of abnormally low HRV in a population of post-infarction patients in respect to the cut-points chosen (raw or age-adjusted). METHODS: Normal population of 296 healthy subjects (81 f, 215 m, aged 47+/-10 years) and post-infarction population of 298 patients (>3 months after acute MI, 65 f, 233 m, aged 56+/-10) were examined. The normal population was divided into 4 subsets based on age at entry: <35, 35-44, 45-54 and >54 years. Based on a 24 h ECG the standard HRV analysis was performed to obtain the following indices: mean RR interval, SDNN and SDANN. A new index of HRV, HRV fraction (HRVF, %) was calculated based on a numerical processing of the RR intervals return map. All patients were followed for 24 months. The endpoints of the study were death (of any reason) and cardiac death. RESULTS: Means and normal limits for SDNN, SDANN and HRVF were: 147+/-36 ms [95% CI 89-220], 136+/-36 ms [79-212] and 53+/-9% [35-68]. The HRV values below the lower normal limit (LNL) were observed in 20-25% of post-MI patients. During a 2 year follow-up there were 36 deaths (total mortality 12.1%), while cardiac mortality was 9.1% (27 cases). The prognostic value of the analysed indices was similar (sens approximately 53-61%, spec approximately 79-84%, PPV 22-26%, NPV 93-94%) irrespective of the cut-points chosen (calculated either for the entire population or age-related). Multivariate Cox regression analysis showed that a decrease of any index below the LNL was associated with a approximately 2.5 and approximately 4-6 times greater risk for death and cardiac death, irrespective of the cut-points chosen. CONCLUSIONS: The age-dependence of the HRV indices does not seem to significantly influence their prognostic value. Thus, a single cut-point of a particular HRV index, based on the entire population, is sufficient to be treated as a risk predictor. In the late phase of myocardial infarction the value of any global HRV index lying below the lower normal limit indicates independently an increased risk of death, especially cardiac death. The new index (HRV fraction) seems to be a promising substitute for currently used standard indices.