Neuronal mTORC1 inhibition promotes longevity without suppressing anabolic growth and reproduction in C. elegans.

mTORC1 (mechanistic target of rapamycin complex 1) is a metabolic sensor that promotes growth when nutrients are abundant. Ubiquitous inhibition of mTORC1 extends lifespan in multiple organisms but also disrupts several anabolic processes resulting in stunted growth, slowed development, reduced fertility, and disrupted metabolism. However, it is unclear if these pleiotropic effects of mTORC1 inhibition can be uncoupled from longevity. Here, we utilize the auxin-inducible degradation (AID) system to restrict mTORC1 inhibition to C. elegans neurons. We find that neuron-specific degradation of RAGA-1, an upstream activator of mTORC1, or LET-363, the ortholog of mammalian mTOR, is sufficient to extend lifespan in C. elegans. Unlike raga-1 loss of function genetic mutations or somatic AID of RAGA-1, neuronal AID of RAGA-1 robustly extends lifespan without impairing body size, developmental rate, brood size, or neuronal function. Moreover, while degradation of RAGA-1 in all somatic tissues alters the expression of thousands of genes, demonstrating the widespread effects of mTORC1 inhibition, degradation of RAGA-1 in neurons only results in around 200 differentially expressed genes with a specific enrichment in metabolism and stress response. Notably, our work demonstrates that targeting mTORC1 specifically in the nervous system in C. elegans uncouples longevity from growth and reproductive impairments, and that many canonical effects of low mTORC1 activity are not required to promote healthy aging. These data challenge previously held ideas about the mechanisms of mTORC1 lifespan extension and underscore the potential of promoting longevity by neuron-specific mTORC1 modulation.

Predictors of cardiac arrest in severe accidental hypothermia.

STUDY OBJECTIVE: To indicate predictors of witnessed hypothermic cardiac arrest. METHODS: We conducted a retrospective analysis of 182 patients with severe accidental hypothermia (i.e., with core body temperature of ≤28 °C) who presented with preserved spontaneous circulation at first contact with medical services. We divided the study population into two groups: patients who suffered hypothermic cardiac arrest (HCA) at any time between encounter with medical service and restoration of normothermia, and those who did not sustain HCA. The analyzed outcome was the occurrence of cardiac arrest prior to achieving normothermia. Hemodynamic and biochemical parameters were analyzed with regard to their association with the outcome. RESULTS: Fifty-two (29%) patients suffered HCA. In a univariable analysis, four variables were significantly associated with the outcome, namely heart rate (p < 0.001), systolic blood pressure (p = 0.03), ventricular arrhythmia (p = 0.001), and arterial oxygen partial pressure (p = 0.002). In the multivariable logistic regression the best model predicting HCA included heart rate, PaO2, and Base Excess (AUROC = 0.78). In prehospital settings, when blood gas analysis is not available, other multivariable model including heart rate and occurrence of ventricular arrhythmia (AUROC = 0.74) can be used. In this study population, threshold values of heart rate of 43/min, temperature-corrected PaO2 of 72 mmHg, and uncorrected PaO2 of 109 mmHg, presented satisfactory sensitivity and specificity for HCA prediction. CONCLUSIONS: In patients with severe accidental hypothermia, the occurrence of HCA is associated with a lower heart rate, hypoxemia, ventricular arrhythmia, lower BE, and lower blood pressure. These parameters can be helpful in the early selection of high-risk patients and their allocation to extracorporeal rewarming facilities.

MicroRNA composition of plasma extracellular vesicles: a harbinger of late cardiotoxicity of doxorubicin.

BACKGROUND: The use of doxorubicin is associated with an increased risk of acute and long-term cardiomyopathy. Despite the constantly growing number of cancer survivors, little is known about the transcriptional mechanisms which progress in the time leading to a severe cardiac outcome. It is also unclear whether long-term transcriptomic alterations related to doxorubicin use are similar to transcriptomic patterns present in patients suffering from other cardiomyopathies. METHODS: We have sequenced miRNA from total plasma and extracellular vesicles (EVs) from 66 acute lymphoblastic leukemia (ALL) survivors and 61 healthy controls (254 samples in total). We then analyzed processes regulated by differentially expressed circulating miRNAs and cross-validated results with the data of patients with clinically manifested cardiomyopathies. RESULTS: We found that especially miRNAs contained within EVs may be informative in terms of cardiomyopathy development and may regulate pathways related to neurotrophin signaling, transforming growth factor beta (TGFß) or epidermal growth factor receptors (ErbB). We identified vesicular miR-144-3p and miR-423-3p as the most variable between groups and significantly correlated with echocardiographic parameters and, respectively, for plasma: let-7g-5p and miR-16-2-3p. Moreover, vesicular miR-144-3p correlates with the highest number of echocardiographic parameters and is differentially expressed in the circulation of patients with dilated cardiomyopathy. We also found that distribution of particular miRNAs between of plasma and EVs (proportion between compartments) e.g., miR-184 in ALL, is altered, suggesting changes within secretory and miRNA sorting mechanisms. CONCLUSIONS: Our results show that transcriptomic changes resulting from doxorubicin induced myocardial injury are reflected in circulating miRNA levels and precede development of the late onset cardiomyopathy phenotype. Among miRNAs related to cardiac function, we found vesicular miR-144-3p and miR-423-3p, as well as let-7g-5p and miR-16-2-3p contained in the total plasma. Selection of source for such studies (plasma or EVs) is of critical importance, as distribution of some miRNA between plasma and EVs is altered in ALL survivors, in comparison to healthy people, which suggests that doxorubicin-induced changes include miRNA sorting and export to extracellular space.

Impact of bisphenol-A and synthetic estradiol on brain, behavior, gonads and sex hormones in a sexually labile coral reef fish.

Endocrine disrupting chemicals, such as bisphenol A (BPA) and ethinylestradiol (EE2), are detected in the marine environment from plastic waste and wastewater effluent. However, their impact on reproduction in sexually labile coral reef fish is unknown. The objective of this study was to determine impacts of environmentally relevant concentrations of BPA and EE2 on behavior, brain gene expression, gonadal histology, sex hormone profile, and plasma vitellogenin (Vtg) levels in the anemonefish, Amphiprion ocellaris. A. ocellaris display post-maturational sex change from male to female in nature. Sexually immature, male fish were paired together and fed twice daily with normal food (control), food containing BPA (100 µg/kg), or EE2 (0.02 µg/kg) (n = 9 pairs/group). Aggression toward an intruder male was measured at 1, 3, and 6 months. Blood was collected at 3 and 6 months to measure estradiol (E2), 11-ketotestosterone (11-KT), and Vtg. At the end of the study, fish were euthanized to assess gonad morphology and to measure expression of known sexually dimorphic genes in the brain. Relative to control, BPA decreased aggression, altered brain transcript levels, increased non-vitellogenic and vitellogenic eggs in the gonad, reduced 11-KT, and increased plasma Vtg. In two BPA-treated pairs, both individuals had vitellogenic eggs, which does not naturally occur. EE2 reduced 11-KT in subordinate individuals and altered expression of one transcript in the brain toward the female profile. Results suggest BPA, and to a lesser extent EE2, pollution in coral reef ecosystems could interfere with normal reproductive physiology and behavior of the iconic sexually labile anemonefish.

Prognostic Factors for Nonasphyxia-Related Cardiac Arrest Patients Undergoing Extracorporeal Rewarming - HELP Registry Study.

OBJECTIVE: Extracorporeal rewarming is the treatment of choice for patients who had hypothermic cardiac arrest, allowing for best neurologic outcome. The authors' goal was to identify factors associated with survival in nonasphyxia-related hypothermic cardiac arrest patients undergoing extracorporeal rewarming. DESIGN: All 38 cardiac surgery departments in Poland were encouraged to report consecutive hypothermic cardiac arrest patients treated with extracorporeal life support. All variables collected were analyzed in order to compare survivor and nonsurvivor groups. The parameters available at the initiation of extracorporeal rewarming were considered as potential predictors of survival in a logistic regression model. The primary outcome was survival to discharge from the intensive care unit. The secondary outcome was neurologic status. SETTING: Multicenter retrospective study. PARTICIPANTS: Ninety-eight cases in the final analysis. INTERVENTIONS: All patients in nonasphyxia-related hypothermic cardiac arrest rewarmed with extracorporeal life support. MEASUREMENTS AND MAIN RESULTS: The survival rate was 53.1%, and 94.2% of survivors had favorable neurologic outcome. The lowest reported core temperature with cerebral performance category scale 1 was 11.8°C. A univariate analysis identified 3 variables associated with survival, namely: age, initial arterial pH, and lactate concentration. In a multivariate analysis, 2 independent predictors of survival were age (0.957; 95% confidence interval [CI] 0.924-0.991) and lactates (0.871; 95% CI 0.789-0.961). The area under the receiver operating characteristics curve for this fitted model was 0.71; 95% CI 0.602-0.817. CONCLUSIONS: Favorable survival with good neurologic outcome in nonasphyxiated hypothermic patients treated with extracorporeal life support was reported. Age and initial lactate level are independently associated with survival.

Decreased number of regulatory T lymphocytes is related to inflammation and number of CD8+ T cells expressing programmed cell death protein-1 in common variable immunodeficiency.

Common variable immunodeficiency (CVID) is a primary immunodeficiency disorder related to recurrent infections, as well as a range of non-infectious manifestations including autoimmune and inflammatory disorders. We hypothesized that patients with CVID and different clinical phenotypes would demonstrate alterations in lymphocyte T subsets, including T lymphocytes expressing programmed cell death protein 1 (PD-1), and regulatory T lymphocytes. We performed flow cytometry in two CVID groups: group 1 with infections only, and group 2 with infections and concomitant noninfectious manifestations. Patients were 18-59 years old (mean 35.8 years of age). Increased proportions of CD8+PD-1+ T cells and reduced regulatory T cells were associated with lymphadenopathy. Amount of regulatory T cells correlated with CD8+PD-1+ T lymphocytes (r = 0.54; p = 0.013), and with CRP (r = -0.64; p = 0.004). Forty percent of patients expressed manifestations in addition to infections (group 2), and they had reduction in number of regulatory T cells [8 (3-12) vs. 24 (11-26)/µl; p = 0.034), naive CD4+ T lymphocytes [36 (27-106) vs. 149 (81-283)/µl; p = 0.034], and elevated C-reactive protein (CRP) [5.33 (3.15-8.82) vs. 1 (1-2.16) mg/l; p = 0.003] in comparison to group 1. In conclusion, the amount of CD8+ T cells expressing PD-1 is associated with lymphadenopathy and number of regulatory T cells in patients with CVID. Patients with CVID and non-infectious complications have increased level of inflammation and alterations in regulatory T cells.

Active feminization of the preoptic area occurs independently of the gonads in Amphiprion ocellaris.

Sex differences in the anatomy and physiology of the vertebrate preoptic area (POA) arise during development, and influence sex-specific reproductive functions later in life. Relative to masculinization, mechanisms for feminization of the POA are not well understood. The purpose of this study was to induce sex change from male to female in the anemonefish Amphiprion ocellaris, and track the timing of changes in POA cytoarchitecture, composition of the gonads and circulating sex steroid levels. Reproductive males were paired together and then sampled after 3 weeks, 6 months, 1 year and 3 years. Results show that as males change sex into females, number of medium cells in the anterior POA (parvocellular region) approximately double to female levels over the course of several months to 1 year. Feminization of gonads, and plasma sex steroids occur independently, on a variable timescale, up to years after POA sex change has completed. Findings suggest the process of POA feminization is orchestrated by factors originating from within the brain as opposed to being cued from the gonads, consistent with the dominant hypothesis in mammals. Anemonefish provide an opportunity to explore active mechanisms responsible for female brain development in an individual with male gonads and circulating sex steroid levels.

Possible sensor applications of selected DNA-surfactant complexes.

Although much research has been performed on DNA complexes carrying long alkyl chains (C10, C16, and C18), there is no information about physicochemical characterization of synthesized composites with allyl imidazole-based ionic liquids and quaternary ammonium salts with n-butyl chains. Here, complexes were synthesized by ion-exchange reactions between sonicated DNA and three ionic liquids (ILs) formed from two imidazole-based compounds, 1-allyl-3-methylimidazolium bromide (Amim) or 1-butyl-3-methylimidazolium bromide (Bmim), and from the quaternary ammonium salt tetra-n-butylammonium bromide (TBAB). Signals in UV-Vis, IR, and CD spectra indicating inclusion of small molecules into the DNA structure confirmed the formation of DNA complexes. Both IR and CD spectra indicated that the B-form conformation of the DNA did not change after the formation of the complexes. Similarly, X-ray diffraction patterns revealed that the formation of IL-DNA complexes did not change the structure of native B-form DNA. Molecular weight (Mw) and radii of gyration (Rg) values of IL-DNA complex chains, established by high-performance size exclusion chromatography coupled with multiangle-laser light-scattering with a differential refractive index detector, were significantly lower than those values found for native DNA molecules due to DNA fragmentation by sonication during complex formation and the direct effects of the IL on the DNA. Scanning electron microscopy images indicate the formation of nanofibres in DNA-Amim and DNA-Bmim complexes, whereas the formation of nanowires was found in samples of DNA-TBAB complexes. Changes in optical properties confirmed by UV and photoluminescence make DNA-IL complexes potential candidates for biosensor application.

A mixture of cellulose production waste with municipal sewage as new material for an ecological management of wastes.

Chemical contaminants present in cellulose production waste and municipal sewage sludge condition the necessity to treat these wastes before they are introduced into the environment. Environmental use of the these wastes appears to be justified owing to the content of organic substances as well as the considerable fertilising value. The aim of the study was to assess chemical composition and ecotoxicity of cellulose production waste and municipal sewage sludge in terms of their eco-friendly application. The assessment was done based on the established soil environment according to regulations in Poland. The research on the chemical composition of the wastes was conducted under laboratory conditions, determining the content of macroelements, microelements and heavy metals with respect to using these wastes in the environment. Phytotoxkit and Microtox tests were performed in order to determine the ecotoxicity of the wastes in the aspect of their environmental use. It was established that, compared to the cellulose production waste, the municipal sewage sludge used in the incubation experiment had a higher content of macroelements and microelements. The content of heavy metals in the waste did not exceed the permissible content that conditions their use in agriculture and reclamation. Based on the results of bioassays it was established that the cellulose production waste was toxic, whereas the combination of cellulose production waste and sewage sludge was found to be of low toxicity to test organisms. The applied wastes and their mixtures were ranked according to phytotoxicity in the following order: cellulose production waste > cellulose production waste + soil > cellulose production waste + sewage sludge. The research results indicate that waste mixtures qualify for environmental use. With respect to the variable chemical composition of cellulose production waste and municipal sewage sludge as well as their ecotoxicity, it was established that each batch intended for environmental use must be subjected to chemical analysis and ecotoxicological tests.

Serum levels of unique miR-551-5p and endothelial-specific miR-126a-5p allow discrimination of patients in the early phase of acute pancreatitis.

OBJECTIVES: Vascular dysfunction is a severe complication which can cause organ ischemia and damage during acute pancreatitis (AP). Laboratory assessment of AP is based on several routine parameters and does not reflect endothelial dysfunction or organ injury. Recently, small non-protein-coding RNAs (miRNAs) have been introduced to laboratory diagnostics as new biomarkers or predictive parameters. Candidate miRNAs (hsa-miR-16-5p, -103a-3p, -122-5p, -126-5p, -148a-5p, -216a-5p, -375, and -551b-5b) were selected to check their possible clinical application in stratification of patients with different AP severity. METHODS: In this observational study, 62 patients with mild (MAP) and 26 with moderate and severe (SAP) acute pancreatitis were included. The control group consisted of 10 age and sex matched subjects. Circulating miRNAs were analyzed in serum using a quantitative real-time PCR method (q-RT-PCR) by means of 3'-locked-nucleic-acid primers. RESULTS: In SAP patients, a significant increase in most of the selected miRNAs (miR-126-5p, -148a-3p, -216a-5p and -551b-5p, and miR-375) was observed when compared to control subjects. In MAP patients, three miRNAs were significantly elevated: endothelial-specific miR-216a-5p, -551b-5p, as well as miR-375 that is highly abundant in pancreas. ROC analysis revealed that miR-126-p and miR-551b-5p can predict AP severity (AUC 0.748, sensitivity 60.0%, specificity 87.1%, p < 0.001) and (AUC 0.716; sensitivity 69.2%, specificity 72.6%, p < 0.001). miR-375 was not relevant (AUC 0.458; sensitivity 55.%, specificity 44.4%). CONCLUSIONS: A pancreatic miRNA signature can be useful for assessment of pancreatic injury in the acute phase of AP. Endothelial dysfunction during AP is reflected by levels of specific circulating miRNAs and may help in patient stratification.

Edible chitosan-alginate based coatings enriched with turmeric and oregano additives: Formulation, antimicrobial and non-cytotoxic properties.

In our study we developed the edible chitosan and alginate coatings with turmeric or oregano additives. The objective of the research was to evaluate the dose-dependent cytotoxicity of films. In cell line studies on HepG2 and BJ cells, they were shown to be non-cytotoxic materials (IC50% was not reached). For HepG2 increase in cell proliferation was observed for 3, 4, and 7 mg/mL of OS3 (124,79±9,21; 162,4±10,46; 165,37±18,44) after 72 h. In BJ cells, no significant decrease in proliferation was noted after 24- and 48-hour exposure to OS0 and OS1 (1-7 mg/ml). The addition of oregano (1% v/v) resulted in films with higher elongation at break and 40% higher tensile strength compared to the base (OS0) film. Use of additives significantly increased the thermal stability of the complexes (by an average of 10 °C). Coatings were tested on tofu and had proven potent antimicrobial properties.

Modifications of geldanamycin via CuAAC altering affinity to chaperone protein Hsp90 and cytotoxicity.

Functionalization of alkyne (1) and azide (2) derivatives of geldanamycin (GDM) via dipolar cycloaddition CuAAC yielded 35 new congeners (3-37) with C(17)-triazole arms bearing caps of different nature (basic vs. acidic, hydrophilic vs. hydrophobic). Confrontation of biological data (anticancer activity vs. toxicity in normal cells) with lipophilicity (clogP), dissociation constants (Kd) of complexes with Hsp90 and binding modes to Hsp90 revealed SAR in specific subgroups of GDM derivatives. The most potent GDM congeners 14-16, bearing C(17)-triazole-benzyl-halogen arms exhibited the most optimal clogP values of 2.7-3.1 at favourable binding to Hsp90 (KdHsp90 at µM level). The anticancer activity of 14-16 (IC50 = 0.23-0.41 µM) is higher than those of GDM (IC50 = 0.58-0.64 µM) and actinomycin D (ActD, IC50 = 0.62-0.71 µM) in SKBR-3, SKOV-3 and PC-3 cell lines, with a comparable cytotoxicity in healthy cells. The relationship between structure and attractive anticancer potency (IC50 = 0.53-0.74 µM) is also observed for congeners with C(17)-triazole-saccharide or C(17)-triazole-unsaturated arms. In the former, the absolute configuration at C(4) (ᴅ-glucose vs. ᴅ-galactose) whereas in the latter the length of the unsaturated arm influences the cytotoxic effects due to different binding strength (Kd, ΔE) and modes with Hsp90. Among all triazole congeners of GDM that are biologically attractive and exhibit lower toxicity in normal cells than GDM and ActD, the derivative 22, bearing the C(17)-triazole-cinnamyl arm, shows the lowest Kd (Hsp90), optimal clogP = 2.82, the best pro-apoptotic properties in SKBR-3 and SKOV-3 and the best selectivity indices (SI). For the most potent GDM derivatives with C(17)-triazole arm, the docking studies have suggested the importance of the intermolecular stabilization between the arm and the D57 or Y61 of Hsp90.

Extracorporeal Rewarming Is Associated With Increased Survival Rate in Severely Hypothermic Patients With Preserved Spontaneous Circulation.

Treatment recommendations for rewarming patients in severe accidental hypothermia with preserved spontaneous circulation have a weak evidence due to the absence of randomized clinical trials. We aimed to compare the outcomes of extracorporeal versus less-invasive rewarming of severely hypothermic patients with preserved spontaneous circulation. We conducted a multicenter retrospective study. The patient population was compiled based on data from the HELP Registry, the International Hypothermia Registry, and a literature review. Adult patients with a core temperature <28°C and preserved spontaneous circulation were included. Patients who underwent extracorporeal rewarming were compared with patients rewarmed with less-invasive methods, using a matched-pair analysis. The study population consisted of 50 patients rewarmed extracorporeally and 85 patients rewarmed with other, less-invasive methods. Variables significantly associated with survival included: lower age; outdoor cooling circumstances; higher blood pressure; higher PaCO 2 ; higher BE; higher HCO 3 ; and the absence of comorbidities. The survival rate was higher in patients rewarmed extracorporeally ( p = 0.049). The relative risk of death was twice as high in patients rewarmed less invasively. Based on our data, we conclude that patients in severe accidental hypothermia with circulatory instability can benefit from extracorporeal rewarming without an increased risk of complications.

The Influence of Aminoalcohols on ZnO Films' Structure.

Preparing structures with the sol-gel method often requires control of the basal plane of crystallites, crystallite structures, or the appearance of the voids. One of the critical factors in the formation of a layer are additives, such as aminoalcohols, which increase the control of the sol formation reaction. Since aminoalcohols differ in boiling points and alkalinity, their selection may play a significant role in the dynamics of structure formation. The main aim of this work is to examine the properties of ZnO layers grown using different aminoalcohols at different concentration rates. The layers were grown on various substrates, which would provide additional information on the behavior of the layers on a specific substrate, and the mixture was annealed at a relatively low temperature (400 °C). The research was conducted using monoethanolamine (MEA) and diethanolamine (DEA). The aminoalcohols were added to the solutions in equal concentrations. The microscopic image of the structure and the size of the crystallites were determined using micrographs. X-ray diffractometry and Raman spectroscopy were used for structural studies, phase analysis and to establish the purity of the obtained films. UV-vis absorption and photoluminescence were used to evaluate structural defects. This paper shows the influence of the stabilizer on the morphology of samples and the influence of the morphology and structure on the optical properties. The above comparison may allow the preparation of ZnO samples for specific applications.

Structural changes of different starches illuminated with linearly polarised visible light.

Aqueous suspensions (30% w/w) of spelt, amaranth and wheat starches were illuminated with linearly polarised visible light for 5, 15, 25 and 50 h. For native and illuminated samples, the weight-average molecular weight, the radii of gyration of the starch polysaccharide chains, and the distribution of the amylopectin structural units of the illuminated starches were determined. The susceptibility to α-amylolysis together with the iodine-binding properties and crystalline structure were studied for each of the starch samples. Illumination with linearly polarised visible light indicated changes in the crystalline structure of the polysaccharide chains and led to an increased enzymatic hydrolysis rate constant for the first stage of hydrolysis and final hydrolysis extent. Changes in the molecular structure of the starch indicate that illumination of the starches induced depolymerisation-repolymerisation reactions of starch polysaccharide chains. That rearrangements of the molecular starch structure depend on the illumination time and the botanical source of the starch.

Impact of rescue collapse on mortality rate in severe accidental hypothermia: A matched-pair analysis.

AIM: To assess the impact of the occurrence of cardiac arrest associated with initial management on the outcome of severely hypothermic patients who were rewarmed with Extracorporeal Life Support (ECLS). METHODS: We collected the individual data of patients in a state of severe accidental hypothermia who were found with spontaneous circulation and rewarmed with ECLS, from cardiac surgery departments. Patients were divided into two groups: those with a subsequent cardiac arrest (RC group); and those with the retained circulation (HT3 group), and compared by using a matched-pair analysis. The mortality rates and the neurological status in survivors were compared as the main outcomes. The difference in the risk of death between the HT3 and RC groups was calculated. RESULTS: A total of 124 patients were included into the study: 45 in the HT3 group and 79 in the RC group. The matched cohorts consisted of 45 HT3 patients and 45 RC patients. The mortality rate in both groups was 24% and 49% (p = 0.02) respectively; the relative risk of death was 2.0 (p = 0.02). ICU length of stay was significantly longer in the RC group (p < 0.001). Factors associated with survival in the HT3 group included patient age, rewarming rate, and blood BE; while in the RC group, patient age and lactate concentration. CONCLUSIONS: The occurrence of rescue collapse is linked to a doubling of the risk of death in severely hypothermic patients. Procedures which are known as potential triggers of rescue collapse should be performed with special attention, including in conscious patients.

Cranial Irradiation in Childhood Acute Lymphoblastic Leukemia Is Related to Subclinical Left Ventricular Dysfunction and Reduced Large Artery Compliance in Cancer Survivors.

Long-term survivors of acute lymphoblastic leukemia (ALL), the most common childhood malignancy, are at remarkably increased risk of heart failure (HF) in middle age, most likely due anthracycline cardiotoxicity. The role of cranial radiation therapy (CRT) in the development of left ventricular (LV) dysfunction, a predecessor of overt HF, remains unclear. Our aim was to compare LV function and systemic arterial properties according to past CRT in young adult survivors of anthracycline-treated ALL. We studied young adult survivors of childhood ALL at a median of 16 years from diagnosis treated with anthracycline-based chemotherapy, with (n = 12) or without (n = 30) CRT. In addition to fractional shortening (FS) and ejection fraction (EF), LV function was quantified by tissue Doppler imaging of the mitral annulus. Aortic strain/distensibility and arterial compliance were derived from echocardiography and simultaneously recorded pulse pressure. Despite similar FS and EF, peak mitral annular systolic velocity (median (interquartile range): 9.0 (7.5-10.0) vs. 10.0 (8.8-11.5) cm/s, p = 0.05), and early diastolic velocity (13.8 (13.0-14.8) vs. 15.5 (14.0-17.3), p = 0.01) were decreased after chemotherapy combined with CRT compared to chemotherapy without CRT. Systemic arterial compliance was lower in post-CRT subjects (1.0 (0.8-1.2 vs. 1.4 (1.1-1.7) mL/mmHg, p = 0.002). Aortic strain and distensibility were similar regardless of prior CRT. In conclusion, lower arterial compliance and subclinical LV dysfunction may be possible late consequences of past CRT in adult survivors of childhood ALL. Whether arterial stiffening is associated with future HF development in CRT-exposed ALL survivors remains to be investigated.

Evaluation of molecular diagnostic approaches for the detection of BRAF p.V600E mutations in papillary thyroid cancer: Clinical implications.

Differentiated papillary thyroid cancer (PTC) is the most common cancer of the endocrine system. PTC has a very good prognosis and a high 5 year survival rate; however, some patients are unresponsive to treatment, and their diagnosis eventually results in death. Recent efforts have focused on searching for prognostic and predictive factors that may enable treatment personalization and monitoring across the course of the disease. The presence of the BRAF mutation is considered to contribute to the risk of poor clinical course, according to American Thyroid Association (ATA) recommendations. The method used for genotyping can impact the predicted mutation frequency; however, ATA recommendations do not address this issue. We evaluated the molecular diagnostic (BRAF p.V600E mutation) results of 410 patients treated for PTC. We thoroughly analyzed the impact of three different BRAF mutation detection methods, Sanger Sequencing (Seq), allele-specific amplification PCR (ASA-PCR), and quantitative PCR (qPCR), on the frequency of mutation detection in 399 patients. Using Seq, we detected the BRAF mutation in 37% of patients; however, we were able to detect BRAF mutations in 57% and 60% of patients using the more sensitive ASA-PCR and qPCR technologies, respectively. Differences between methods were particularly marked in the thyroid papillary microcarcinoma group; BRAF p.V600E mutations were found in 37% of patients using Seq and 63% and 66% of patients using ASA-PCR and qPCR, respectively. We also evaluated how these different diagnostic methods were impacted by DNA quality. Applying methods with different sensitivities to the detection of BRAF p.V600E mutations may result in different results for the same patient; such data can influence stratification of patients into different risk groups, leading to alteration of treatment and follow-up schemes.

The usefulness of determining the presence of BRAF V600E mutation in fine-needle aspiration cytology in indeterminate cytological results.

INTRODUCTION: Fine-needle aspiration biopsy (FNAB) is regarded as the gold standard method for the diagnosis of thyroid nodules, but it has its limitations. Additional methods that would improve sensitivity and specificity in the diagnosis of thyroid cancer (TC), especially in indeterminate lesions. Molecular tests seem to be such a tool. BRAF V600E mutation (the most common in TC) can be detected in FNAB and can be potentially a very useful ancillary marker for FNAB practice. The aim of our study was to evaluate the usefulness of the detection of the BRAF V600E mutation in FNAC in the early diagnosis of TC in patients with indeterminate cytology. MATERIAL AND METHOD: 2290 FNAB were performed and 147 indeterminate results (group 3, 4, and 5 of the Bethesda system) were obtained. Material from these groups was submitted for molecular tests for the occurrence of BRAF V600E mutation. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the tests were calculated. RESULTS: Determining the presence of BRAF V600E mutation in FNAC material in groups 3 and 4 together and in group 5 is associated with sensitivity of TC diagnosis of 37.5% and 81.8%, respectively. In all cases the detection of BRAF V600E mutation was associated with histopathologically proving the presence of TC (specificity of the test - 100%). CONCLUSIONS: The presence of BRAF V600E mutation in FNAC material is always associated with the presence of TC. The usefulness of determining the presence of BRAF V600E in FNAC in cytological groups 3 and 4 is associated with low sensitivity in the diagnosis of thyroid cancer. Due to its high specificity BRAF V600E study may be useful in determining the scope of surgery in patients in cytological group 5.