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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., (© 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.)

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.

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.

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., (© 2017 John Wiley & Sons Ltd.)

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.

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., (Copyright 2002 European Society of Cardiology)

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.

Objective: QT interval prolongation and increased spatial QT dispersion are important factors increasing the risk in coronary heart disease. The authors studied the spontaneous beat-to-beat variability of ventricular repolarization (RT intervals) in normal subjects and in patients after myocardial infarction (MI) in order to define the determinants of abnormal temporal dispersion., Methods: Seventy-six patients with a history of MI (17 female, 59 male, aged 52 +/- 10 years) comprised the study group. Forty-seven patients had preserved left ventricular ejection fraction (EF > or = 40%, MI-A) and 29 patients had left ventricular dysfunction (EF < 40%, MI-B). Twenty healthy volunteers (6 female, 14 male, aged 25 +/- 5 years) were included as the control group. An ECG signal of 512 heartbeats was recorded in the supine position. After analogue-to-digital conversion (16 bit, 2 kHz), the fiducial points of the R wave and T wave were determined. The RR and RT variability (V) assessed in the time domain as the standard deviations of RR and RT (ms), as well as the coefficients of scatterplots of RR and RT intervals., Results: As expected, the standard deviation of RR was significantly reduced in MI patients. The magnitude of RTV in the time domain was similar in the controls and in both subgroups of MI patients. The complexity of heart rate variability (HRV) was slightly, but significantly, reduced in the MI-B group, but not significantly in the MI-A heart group. The complexity of RTV behaved in the opposite manner, being increased in both MI subgroups with the lower mean in the MI-B patients. The different behaviour of HRV and RTV was indicated by the increased ratio of RR/RT coefficients, which reached a significantly greater value in the MI-B group., Conclusion: The authors have described different patterns of scatterplot of short-term HRV and RTV in normal subjects, which confirmed that RTV is a less complex phenomenon than HRV. In patients after MI, the complexity of HRV diminishes, while the complexity of RTV increases. These opposing changes are more pronounced in patients with left ventricular dysfunction. A possible prognostic value of this feature is unknown and remains to be elucidated in future prospective studies.

Background and Study Aims: Cardiopulmonary complications have been reported during upper gastrointestinal endoscopy and endoscopic retrograde cholangiopancreatography (ERCP). The aim of this study was to evaluate the sympathovagal response to the stretching of the common bile duct caused by contrast medium injection during ERCP., Patients and Methods: The 16 patients included in the study were assigned to two groups according to the size of common bile duct. Group A consisted of patients with normal-sized bile ducts, while Group B patients had dilated common bile ducts. The heart rate variability (HRV) technique was used to assess the sympathovagal balance. The HRV data were gathered using the Holter technique, and frequency domain analysis revealed two main spectral components of HRV: low frequency (LF) and high frequency (HF)., Results: In patients with normal-sized bile ducts, contrast injection initiated a rapid increase in the power spectra (ms2) of both the HF and LF components. In patients with dilated common bile ducts, this phenomenon was not observed., Conclusions: During ERCP, contrast injection into a dilated common bile duct does not stimulate the autonomic nervous system in the same way as it does when the common bile duct is of normal size.

One of the coronary disease diagnostic methods of big sensitivity and specificity is perfusive effort scintrigraphy of myocardium by means of thallium-201. For few years the dipyridamole test has been applied instead of the effort test. Perfusive scintigraphy of myocardium after provocative treatment by means of dipyridamole and then selective coronary arteriography of coronary vessels, have been carried out at 25 patients with ischemia. These studies showed almost 100% of conformability with exposing the ischemia zones in scintigraphy and coronary arteriography in cases of coronary vessels contraction over 50%. The dipyridamole test also revealed the ischemzones in myocardium that are in agreement with coronary vascularization deficiency. That test can be utilized in revealing the coronary disease and is helpful in qualifying patients for coronary arteriography. At the same time studies that have been carried out prove that applying dipyridamole is absolutely contraindicated in treating of coronary disease of the organic background.

Sudden cardiac death caused by cardiac arrhythmia's or asystolies in patients with coronary heart disease can often be avoided if resuscitation is administered immediately, preferably before the patient loses consciousness. In cases when rapid help is not available usually death occurs. We have studied a method of cardiopulmonary resuscitation (CPR) which can be self--administered by patients trained in recognizing imminent arrival of life-threatening Morgani Adams Stokes (MAS) events. The recent study comprised the three methods of investigation in three separate groups of patients: the first group underwent invasive procedures (20 pts), the second non invasive Doppler studies (31 pts) and the third in-and outhospital clinical observations (115 pts). The results indicate that evoked coughing can effectively prevent fainting and maintaining consciousness until conventional CPR help becomes available.

Hemodynamic changes related to pregnancy in women with Congenital Heart Disease (ConHD) or Acquired Valvular Heart Disease (AVHD) require special medical care during pregnancy and especially immediately prior to, and after delivery. Following a five year retrospective study (1978-1982), the authors monitored prospectively over a period of 12 years (1983-1994), 378 pregnancies in 252 women with ConHD and AVHD. Based on the results of the initial clinical examination and monthly follow-up visits, a numerical risk score was assigned for each patient. The study showed good conformity between the predictive risk score values and the patient's condition in the course of pregnancy and delivery. Thus, this method represents a novel and potentially very useful clinical tool for management of pregnancy in patients with ConHD and AVHD.

64 years old female operated on for right auricular tumor, which manifested as right ventricular heart failure is presented. Although full preoperative diagnostics was performed the tumor appeared to be metastatic one with the primary focus situated in the liver (histologically ca hepatocellular). Necessity of cavography in case of right auricular tumor is stressed in the paper.