Recent Advances in Methods for Circulating Tumor Cell Detection.

Circulating tumor cells (CTCs) are released from primary tumors and transported through the body via blood or lymphatic vessels before settling to form micrometastases under suitable conditions. Accordingly, several studies have identified CTCs as a negative prognostic factor for survival in many types of cancer. CTCs also reflect the current heterogeneity and genetic and biological state of tumors; so, their study can provide valuable insights into tumor progression, cell senescence, and cancer dormancy. Diverse methods with differing specificity, utility, costs, and sensitivity have been developed for isolating and characterizing CTCs. Additionally, novel techniques with the potential to overcome the limitations of existing ones are being developed. This primary literature review describes the current and emerging methods for enriching, detecting, isolating, and characterizing CTCs.

Circulating tumor nucleic acids: biology, release mechanisms, and clinical relevance.

BACKGROUND: Despite advances in early detection and therapies, cancer is still one of the most common causes of death worldwide. Since each tumor is unique, there is a need to implement personalized care and develop robust tools for monitoring treatment response to assess drug efficacy and prevent disease relapse. MAIN BODY: Recent developments in liquid biopsies have enabled real-time noninvasive monitoring of tumor burden through the detection of molecules shed by tumors in the blood. These molecules include circulating tumor nucleic acids (ctNAs), comprising cell-free DNA or RNA molecules passively and/or actively released from tumor cells. Often highlighted for their diagnostic, predictive, and prognostic potential, these biomarkers possess valuable information about tumor characteristics and evolution. While circulating tumor DNA (ctDNA) has been in the spotlight for the last decade, less is known about circulating tumor RNA (ctRNA). There are unanswered questions about why some tumors shed high amounts of ctNAs while others have undetectable levels. Also, there are gaps in our understanding of associations between tumor evolution and ctNA characteristics and shedding kinetics. In this review, we summarize current knowledge about ctNA biology and release mechanisms and put this information into the context of tumor evolution and clinical utility. CONCLUSIONS: A deeper understanding of the biology of ctDNA and ctRNA may inform the use of liquid biopsies in personalized medicine to improve cancer patient outcomes.

Tuneable in-situ nanoCT workflow using FIB/SEM.

Inspired by the standard computed tomography, a new method of 3D X-ray imaging embedded in FIB-SEM microscope is proposed. The unique combination of TEM-like specimen stage enabling in lens STEM detection (referred to as CompuStage), nanomanipulator (referred to as EasyLift) facilitating in-situ sample transfer from bulk sample to TEM-like stage and pixelated in-situ Timepix X-ray detector in Helios G4 FX FIB-SEM system offers an unprecedented workflow. Motivated by common circular CT scan known from microCT world, the object under study is placed on CompuStage rod which enables two possible rotation (in TEM/SEM terminology called tilt) movements - α-tilt - rotation of the CompuStage rod around its axis, and ß-tilt - rotation around axis perpendicular to CompuStage rod. ß-tilt rotation enables a circular movement of the sample while α-tilt sets the correct position of sample with respect to target and detector. Thin metal lamella of suitable material welded to EasyLift manipulator needle is used as an X-ray target. The final target-sample geometry - position, distance - can be fine-tuned using position control of CompuStage and EasyLift and in-situ monitored by SEM. Both sample and target can also be easily prepared in-situ. Radiographs are recorded by Timepix detector with inherent noise-free operation and energy filtration. For the 3D reconstruction standard microCT reconstruction algorithm is used with the procedure adjusted for the format and quality of nanoCT images. The experiments were carried out on Helios G4 FX DualBeam using titanium and tungsten targets and various semiconductor samples. The ultimate resolution of the proposed method in orders of tens of nanometers was achieved both by the possibility of close target to sample positioning and of adjustment of primary beam energy down to low energies reducing the interaction volume in the target. Since the lower energy radiation is well suited for life-science, the method was also tested on several bio-samples using silver target. The silver target, thanks to its massive low energy Lα line, allowed to distinguish subtle structures in the resin embedded stained mouse brain and also to observe and reconstruct canaliculi in the mouse bone (earlier reported by Dierolf et al. 2010, Nature 467 436).

A comprehensive interdisciplinary view at the Return to Sport after COVID-19 infection.

The COVID-19 pandemic has affected the whole world. It applies to all age and social groups. It is no different with athletes. So far, we cannot say for sure what the long-term consequences of SARS-CoV-2 infection are. Recent evidence, however, suggests that we should be very careful when returning to sports. After self-isolation, the athlete should undergo a Preparticipation Physical Examination and then pay attention to the gradual dosing of the load to prevent complications. Lifestyle changes and care for the mental health of athletes are also necessary during the illness. In this work, we present a comprehensive methodology for returning to sports after COVID-19 for medical and coaching teams caring for athletes divided according to the course of the disease. In scientific literature, similar algorithms are called "Return to Play" or "Return to Sport". Creating an exact algorithm can make the Return to Play process more efficient and safer. However, increased attention still needs to be paid to certain organ systems and specific symptoms that could indicate long-term consequences to the new type of coronavirus.

Urine fetuin-A values in relation to the presence of urolithiasis.

OBJECTIVE: To investigate the relationship of urine fetuin-A and other promotors and inhibitors of urine crystalization with urolithiasis, as fetuin-A inhibits the precipitation of hydroxyapatite from supersaturated solutions of calcium and phosphate in vitro but no information on urine fetuin-A in patients with urolithiasis is available. PATIENTS AND METHODS: In all, 39 patients with urolithiasis and 22 individuals with no urolithiasis or probands with undetected stones were involved. All patients underwent kidney ultrasonography and X-ray examination, and body mass index (BMI) was calculated. Serum creatinine, parathyroid hormone, calcium, magnesium, anorganic phosphate, uric acid and urine creatinine, albumin, alpha(1)-microglobulin, sulphate, oxalate, citrate and fetuin-A (ELISA) were determined. RESULTS: The patients with urolithiasis had lower urine fetuin-A levels (median 4.9 vs 0.77 mg/day; P < 0.01) and citraturia levels (1.7 vs 5.1 mmol/day; P = 0.02); and higher calciuria (6.5 vs 5.2 mmol/day) and oxaluria (0.47 vs 0.25; P = 0.04). Patients with fetuin-A levels in the lowest quartile had an odds ratio of 36 compared with individuals in the highest quartile. The sensitivity of the urine fetuin-A level for urolithiasis was 97.4% and specificity was 100% (area under the curve 0.99; 95% confidence interval 0.94-1.0) using a urine fetuin-A threshold of

Assessment of the influence of exercise on heart rate variability in anxiety patients.

OBJECTIVES: The aim of the study was to determine the status of the autonomic nervous system (ANS) in anxiety disorder patients and to evaluate possible exercise intervention in order to improve the ANS and overall psychiatric status of patients. PATIENTS AND METHODS: The ANS function was monitored via means of heart rate variability (HRV) changes during a 6-week hospitalization at the Psychiatric Clinic of University Hospital in Olomouc. The status of ANS was monitored by the new evaluation method of spectral analysis (SA) of HRV. The research involved 43 anxiety patients (29 women and 14 men) who underwent six weeks of intensive psychotherapy; part of the test group also participated in regular aerobic exercise. RESULTS: In the sense of autonomic dysfunction we can not give clear-cut answers to the question whether the autonomic dysfunction could be one of the predictors of anxiety disorder, although our results suggest lower vagal representation in the spectra. Unlike in the healthy population the exercise had a positive impact only on a limited number of patients. Regular endurance exercise on a stationary bicycle had a positive impact on the ANS efficiency only in patients with primary ANS activity reduction. In patients with normal ANS efficiency the exercise intervention had no effect; in some cases we even found activity reduction during the hospitalization period. However, the exercise had a positive effect on the course and outcome of the applied therapy in all patients. CONCLUSIONS: ANS disturbances have not been exhibited in some patients suffering from anxiety disorder. Due to the positive impact of regular physical activity on cardiovascular, metabolic, neural, and psychological changes in organism, monitored exercise should be implemented into therapeutic programs for patients with anxiety disorders.

Short-term spectral analysis of heart rate variability during supine-standing-supine test in patients with paroxysmal atrial fibrillation.

The aim of the study was to assess the sympathovagal balance in group of 27 patients without significant structural heart disease after an attack of atrial fibrillation. The investigation was performed using spectral analysis of heart rate variability during examination under conditions of different orthostatic loads in single phases, called the supine-standing- supine test. The findings were compared with a group of healthy persons. These revealed a significantly decreased total spectral power (430.7 vs 1558.0 ms(2) supine1; 477.6 vs 1042,5 ms(2) standing; 567.5 vs 1948.5 ms(2) supine2), and spectral power of the high frequency spectral component (140.8 vs 619.3 ms(2) supine1; 96.2 vs 203.3 ms(2) standing; 186.3 vs 739.4 ms(2) supine2) in the studied group of patients in comparison with the control group.

Assessment of spectral analysis of heart rate variability in patients with history of atrial fibrillation by means of age-dependent parameters.

Heart rate variability evaluation is a useful diagnostic tool for autonomic nervous balance assessment. The role of the autonomic nervous system in aetiology of atrial fibrillation is sometimes clear as a trigger from a patient's history, but mostly it acts as a modulating factor which is not easy to detect. The present study demonstrates results of spectral analysis of short-term heart rate variability during ortho-clinostatic tests processed by means of age-dependent parameters. An original telemetric system and a unique method for heart rate variability assessment, developed by the Faculty of Physical Culture, were applied for the first time to examine patients with the history of atrial fibrillation.