Erratum - Tri-Lineage Differentiation of NTERA2 Clone D1 Cells towards Neural, Hepatic and Osteogenic Lineages in Vitro.

The images in Fig. 4 were not presented correctly. The correct version of Fig. 4: see the last page of pdf. The original article was published in Folia Biologica (Praha) Volume 67, No. 5-6 (2021), 174-182. https://doi.org/10.14712/fb2021067050174.

The relationship between cognitive control and lexical conflict resolution in developmental dyslexia.

Previous studies have revealed that cognitive control functions contribute to the resolution of lexical interference. Both cognitive control (CC) deficits and reduced speed of lexical retrieval in Rapid Automatized Naming (RAN) tasks are characteristics of Developmental Dyslexia (DD), but it is still not fully understood how these deficits relate to each other and to reading problems. To examine this question, we tested adolescents with DD (n = 38), poor readers (PR; n = 25) and typical readers (TR; n = 33) matched on age and IQ, on CC functions with Stroop, Stop Signal, Simon, Backward Digit Span and n-back tasks and on lexical retrieval and lexical conflict resolution with RAN of pictures in semantically homogeneous vs. mixed trials. As expected, in the blocked RAN Task DD individuals showed longer naming times and a greater effect of lexical conflict resolution (indexed by difference scores of naming times in the homogeneous and mixed conditions) than TR participants. We also found significant group differences (TR = PR > DD) in CC measures. Naming time was associated with CC, while the lexical interference effect did not show any association with this set of abilities. These findings suggest that DD individuals show impairments in multiple cognitive functions, such as cognitive control, lexical retrieval and lexical conflict resolution. Our results also suggest that CC functions are involved in lexical retrieval, but we have not found evidence for their involvement in lexical conflict resolution processes.

Tri-Lineage Differentiation of NTERA2 Clone D1 Cells towards Neural, Hepatic and Osteogenic Lineages in Vitro.

Over the past decades, the in vitro use of pluripotent cell lines gained a crucial role in toxicology, preclinical drug testing and developmental biology. NTERA2 clone D1 cells were identified as pluripotent cells with high potential for neural differentiation. Although they are commonly used cellular sources in neuropharmacology and neurodevelopmental studies, their endodermal and mesodermal differentiation potential awaits further characterization. Here, we devised improved protocols for hepatogenic and osteogenic differentiation of NTERA2 clone D1 cells. Our in vitro differentiation assays showed significant up-regulation of multiple hepatogenic markers. We also observed robust mineralization and osteogenic marker expression of NTERA2 clone D1 cells upon in vitro osteogenic induction. These results suggest that NTERA2 clone D1 cells may be utilized as an in vitro model system to study various aspects of liver biology and osteogenesis. In addition, tri-lineage differentiation of NTERA2 clone D1 cells may serve as a simple experimental control system when validating pluripotency of other cell types.

Next-generation sequencing identifies novel mitochondrial variants in pituitary adenomas.

PURPOSE: Disrupted mitochondrial functions and genetic variants of mitochondrial DNA (mtDNA) have been observed in different human neoplasms. Next-generation sequencing (NGS) can be used to detect even low heteroplasmy-level mtDNA variants. We aimed to investigate the mitochondrial genome in pituitary adenomas by NGS. METHODS: We analysed 11 growth hormone producing and 33 non-functioning [22 gonadotroph and 11 hormone immunonegative] pituitary adenomas using VariantPro™ Mitochondrion Panel on Illumina MiSeq instrument. Revised Cambridge Reference Sequence (rCRS) of the mtDNA was used as reference. Heteroplasmy was determined using a 3% cutoff. RESULTS: 496 variants were identified in pituitary adenomas with overall low level of heteroplasmy (7.22%). On average, 35 variants were detected per sample. Samples harbouring the highest number of variants had the highest Ki-67 indices independently of histological subtypes. We identified eight variants (A11251G, T4216C, T16126C, C15452A, T14798C, A188G, G185A, and T16093C) with different prevalences among different histological groups. T16189C was found in 40% of non-recurrent adenomas, while it was not present in the recurrent ones. T14798C and T4216C were confirmed by Sanger sequencing in all 44 samples. 100% concordance was found between NGS and Sanger method. CONCLUSIONS: NGS is a reliable method for investigating mitochondrial genome and heteroplasmy in pituitary adenomas. Out of the 496 detected variants, 414 have not been previously reported in pituitary adenoma. The high number of mtDNA variants may contribute to adenoma genesis, and some variants (i.e., T16189C) might associate with benign behaviour.

Synthesis, Characterization and Photocatalytic Efficiency of ZnO/MWCNT Nanocomposites Prepared Under Different Solvent Conditions.

Here we report the application of zinc oxide (ZnO) coated multi-walled carbon nanotube (MWCNT) composites in the photocatalytic decomposition of acetaldehyde (AA). Zinc oxide nanoparticles were successfully coated on the multi-walled carbon nanotube via impregnation process using zinc acetate (Zn(CH3COO)2×2H2O) as precursor and sodium dodecyl sulfate (SDS) treated multiwalled carbon nanotube as raw material under different solvent conditions. The applied solvents during preparation were ethanol (EtOH) and water (H2O). As-prepared materials were characterized by thermal analysis (TG), X-ray diffraction (XRD), specific surface area measurement (BET) and transmission electron microscopy (TEM) techniques. Photocatalytic efficiencies of as-prepared composites were investigated in a stationary reactor equipped with UV lamp. Decomposition of acetaldehyde was followed by using gas chromatography (GC). Observations revealed that using impregnation method and different solvents the preparation of ZnO/MWCNT nanocomposites can be controlled easily. The highest degradation rate was achieved with the nanocomposite was synthetized using ethanol as solvent. The photocatalytic experiments revealed that the composite has higher photocatalytic activity than that of both the zinc oxide nanoparticles and the mechanical mixture of multi-walled carbon nanotube and zinc oxide.

Tracking down protein-protein interactions via a FRET-system using site-specific thiol-labeling.

Förster resonance energy transfer is among the most popular tools to follow protein-protein interactions. Although limited to certain cases, site-specific fluorescent labeling of proteins via natural functions by means of chemical manipulations can redeem laborious protein engineering techniques. Herein we report on the synthesis of a heterobifunctional tag and its use in site-specific protein labeling studies aiming at exploring protein-protein interactions. The oxadiazole-methylsulfonyl functionality serves as a thiol specific warhead that enables easy and selective installation of fluorescent labels through a bioorthogonal motif. Mitogen activated protein kinase (MAPK14) and its substrate mitogen activated protein kinase activated kinase (MAPKAP2) or its docking motif, a 22 amino acid-long peptide fragment, were labeled with a donor and an acceptor, respectively. Evolution of strong FRET signals upon protein-protein interactions supported the specific communication between the partners. Using an efficient FRET pair allowed the estimation of dissociation constants for protein-protein and peptide-protein interactions (145 nM and 240 nM, respectively).

Expression of G protein-coupled oestrogen receptor in melanoma and in pregnancy-associated melanoma.

BACKGROUND: The hormone sensitivity of melanoma and the role of 'classical' oestrogen receptor (ER) α and ß in tumour progression have been intensively studied with rather contradictory results. The presence of 'non-classical' G protein-coupled oestrogen receptor (GPER) has not been investigated on human melanoma tissues. OBJECTIVE: To analyse the expression of GPER, ERα and ERß in pregnancy-associated (PAM) and in non-pregnancy-associated (NPAM) melanomas in correlation with traditional prognostic markers and disease-free survival (DFS). METHODS: Receptor protein levels were tested using immunohistochemistry in 81 formalin-fixed paraffin-embedded melanoma tissues. PAMs (n = 38) were compared with age- and Breslow thickness-matched cases (n = 43) including non-pregnant women (NPAM-W) (n = 22) and men (NPAM-M) (n = 21). The association between receptor expression and DFS was analysed by uni- and multivariate Cox proportional hazards regression. RESULTS: G protein-coupled oestrogen receptor was detected both in PAMs and NPAMs. In 39 of the 41 (95.1%) GPER-positive melanomas, GPER and ERß were co-expressed. GPER/ERß-positive melanomas were significantly more common in PAM compared to NPAM (P = 0.0001) with no significant difference between genders (P = 0.4383). In PAMs, the distribution of GPER and ERß was similar (78.4% vs. 81.6%; P = 0.8504), while in NPAM, ERß was the representative ER (60.5% vs. 27.9%; P = 0.0010) without gender difference (59.1% vs. 61.9%). GPER-/ERß-positive melanomas were associated with lower Breslow thickness, lower mitotic rate and higher presence of peritumoral lymphocyte infiltration (PLI) compared to GPER-/ERß-negative cases (P = 0.0156, P = 0.0036 and P = 0.0001) predicting a better DFS (HR = 0.785, 95% CI 0.582-1.058). Despite the significantly higher frequency of GPER and ERß expression in PAM, no significant difference was found in DFS between PAM and NPAM. All but one case failed to show ERα expression. CONCLUSIONS: The presence of GPER and its simultaneous expression with ERß can serve as a new prognostic indicator in a significant subpopulation of melanoma patients.

A systematic study of protein labeling by fluorogenic probes using cysteine targeting vinyl sulfone-cyclooctyne tags.

Fluorescent tagging of proteins via accessible cysteine residues is of paramount importance. In this study, model proteins of interest (mitogen-activated protein kinases) were labeled successfully in native state on their free thiols by direct fluorescence derivatization, or in a sequential manner where conjugation of the site specific linker and the fluorophore is carried out in two steps. To this end we designed and prepared two novel chemical reporters carrying vinyl sulfone as Cys targeting function and cyclooctyne motifs, suitable for subsequent conjugation with fluorogenic azides via copper free strain-promoted azide-alkyne click chemistry. Direct and sequential labeling reaction steps were analyzed by native PAGE, capillary zone electrophoresis and tandem mass spectrometry. The efficiency of tagging was correlated with solvent accessibility of the Cys residues. Our results indicated that conjugation of native proteins by vinyl sulfone linkers was fast and thiol-selective. Subsequent click reaction with fluorogenic dyes generates intensive fluorescence signals and fulfills all requirements of bioorthogonality.

Correction: A systematic study of protein labeling by fluorogenic probes using cysteine targeting vinyl sulfone-cyclooctyne tags.

Correction for 'A systematic study of protein labeling by fluorogenic probes using cysteine targeting vinyl sulfone-cyclooctyne tags' by B. Söveges, et al., Org. Biomol. Chem., 2016, 14, 6071-6078.

The interface between HOPG and 1-butyl-3-methyl-imidazolium hexafluorophosphate.

The interface between highly oriented pyrolytic graphite (HOPG) and 1-butyl-3-metyl-imidazolium hexafluorophosphate (BMIPF6) has been studied using cyclic voltammetry, electrochemical impedance spectroscopy, immersion charge measurements and in situ scanning tunneling microscopy (in situ STM). The results are compared with those obtained with Au(100) in BMIPF6 (Phys. Chem. Chem. Phys., 2011, 13, 11627). The main result is that the high frequency capacitance spectra on the two systems are similar to each other, however at low frequencies some slow interfacial processes cause the appearance of a second capacitance arc on Au(100), which is absent for HOPG. The slow processes are attributed to the rearrangement of the Au surface structure and to the formation of ionic liquid adlayers--these are visualized by in situ STM.

Effects of tampons and menses on the composition and diversity of vaginal microbial communities over time.

OBJECTIVE: To investigate the influence of menses on the vaginal microbiota and determine whether tampons that differ in material composition influence these bacterial communities in different ways. DESIGN: A single-centre trial with randomised, complete block design. SETTING: Procter & Gamble facility. SAMPLE: Seven self-declared healthy, female volunteers of reproductive age. METHODS: Volunteers used a pad and two types of tampons during the study, one product exclusively each month for three sequential menstrual cycles. During menses and once each mid-cycle, vaginal bacterial community composition was characterised by cultivation-independent methods based on pyrosequencing of V1-V2 variable regions of 16S ribosomal RNA genes. MAIN OUTCOME MEASURES: Changes in the species composition, abundance and diversity in vaginal bacterial communities over time and between treatments. RESULTS: The vaginal microbiotas of all seven women were dominated by Lactobacillus spp. at mid-cycle, and the compositions of those communities were largely consistent between cycles. Community dynamic patterns during menses varied considerably and were more or less individualised. In three of the seven women the community diversity during pad use was significantly different from at least one tampon cycle. CONCLUSIONS: Changes in the composition of the vaginal microbiota during menses were common, but the magnitude of change varied between women. Despite these changes, most communities were capable of resuming a composition similar to previous mid-cycle sampling times following menstruation. Overall we conclude that the two tampons tested do not significantly impact the vaginal microbiota in different ways; however, larger studies should be performed to confirm these findings.

Partial absence of superior vena cava in an adult patient. Case report and literature review.

Absence of the superior vena cava (SVC) is a rare variety of vascular anomaly. The purpose of this report is to describe the computed tomography (CT) findings of the partial absence of the SVC without persistent left SVC in a patient with no evidence of congenital cardiovascular disease and no prior history of central venous instrumentation. A 77-year-old woman with a history of colon cancer underwent thoracoabdominal CT imaging because of abdominal pain of uncertain cause. No tumor recurrence was observed. A complicated"investigation" confirmed a thymoid cancer surgery back in 1976, which was accompanied by resection of the SVC and the left brachiocephalic vein because of their invasion. Owing to the absence of the SVC and bilateral brachiocephalic veins, caval hypertension developed in the patient, resulting in the dilation of cavo-caval anastomoses. In addition, new anastomoses were opened. The clinical significance and possible embryogenesis of this anomaly are discussed. The extremely rare condition of the partial absence of the SVC appeared with subcutaneous dilated, tortuous collaterals in an asymptomatic adult patient. This anomaly is becoming clinically more relevant with the increasing use of minimally invasive vascular surgery.

Bone marrow stromal cells inhibit mast cell function via a COX2-dependent mechanism.

BACKGROUND: Mast cells (MCs) have a central role in the induction of allergic inflammation, such as seen in asthma, and contribute to the severity of certain autoimmune diseases, such as rheumatoid arthritis. The MC thus represents an important inflammatory cell, and one which has resisted therapeutic attempts to alter its role in disease. OBJECTIVE: Because bone marrow-derived stromal cells (BMSC, also known as mesenchymal stem cells or MSCs) have been reported to alter allergic inflammation in vivo, we chose to study the interaction between mouse BMSC and mouse bone marrow-derived MCs. METHODS: MC degranulation, cytokine production and chemotaxis were evaluated in vitro following co-culture with BMSCs either in cell contact or a transwell. In addition, MC degranulation was assessed in vivo following administration of BMSCs in a model of passive cutaneous anaphylaxis and a peritoneal degranulation assay. Mechanisms of MC suppression by BMSCs were determined through use of inhibitors or antibodies to COX1, COX2, nitric oxide, indoleamine 2, 3-dioxygenase, EP1-4 receptors, TGF-ß and IL-10. Lastly, we utilized either BMSCs or MCs deficient in COX1, COX2 or EP1-4 receptors to confirm the mechanisms of inhibition of MC function by BMSCs. RESULTS: We discovered that BMSCs will effectively suppress specific MC functions in vitro as well as in vivo. When MCs are cocultured with BMSCs to allow cell-to-cell contact, BMSCs suppressed MC degranulation, pro-inflammatory cytokine production, chemokinesis and chemotaxis. Similarly, MC degranulation within mouse skin or the peritoneal cavity was suppressed following in vivo administration of BMSCs. Further, we found that these inhibitory effects were dependent on up-regulation of COX2 in BMSCs; and were facilitated through the activation of EP4 receptors on MCs. CONCLUSION AND CLINICAL RELEVANCE: These observations support the concept that BMSCs have the ability to suppress MC activation and therefore could be the basis for a novel cell based therapeutic approach in the treatment of MC driven inflammatory diseases.

Adhesive Management of Anterior Tooth Wear in Combination with the Dahl Concept-A 27-Month Observational Case Series.

Localized anterior maxillary tooth wear caused by erosion and attrition with loss of interocclusal space is difficult to manage. This observational case-series study reports six cases with worn anterior dentition treated with labial ceramic and palatal direct resin composite veneers at an increased vertical dimension of occlusion without restoration of unaffected posterior teeth. Thirty-six palatal direct veneers were made in six patients from a nanohybrid resin composite with the help of a wax-up-based template at an increased vertical dimension. After the complete re-establishment of posterior occlusion, 40 labial lithium-disilicate ceramic veneers were fabricated with a mock-up-guided method. The sandwich veneers were evaluated according to the United States Public Health Service (USPHS) criteria after a mean service time of 22.7 months. Re-establishment of posterior contacts as well as subjective patient satisfaction and function were evaluated. The overall success of the labial ceramic veneers was excellent. The quality of the palatal resin composite restorations was found to be good with predominantly "Alpha" scores. The marginal quality (11.1% and 33.3% of integrity and discoloration, respectively) and surface roughness (16.7%) showed small deteriorations indicated by "Beta" scores. The resin composite showed, in general, signs of wear facets which resulted in "Beta" scores in 44.4% of the cases. Posterior contacts re-established firmly within 4 weeks in all cases. Patient satisfaction with esthetics and function was high. The short-term outcome of this non-invasive treatment option is favorable and promising.

Controlling the Structure of Carbon Deposit by Nitrogen Doping Catalytic Chemical Vapor Deposition Synthesis.

Nitrogen doped multi-walled carbon nanotubes and other carbon nanoparticles were synthesized by catalytic chemical vapor deposition of tripropylamine and acetylene on CaCO3-supported cobalt catalyst (5 wt%), prepared by impregnation, and various precursors. Each synthesis was performed by using either the pure nitrogenous organic compound or its mixture with acetone. Transmission electron microscopy studies revealed a significant difference both in the yield and the diversity of the carbon deposits. Every synthesis resulted in bamboo-like nanotubes, and nearly all of them also in onion-like structures. Electron energy loss spectroscopy studies of the samples indicated the presence of nitrogen and calcium (caused by the catalyst support). High-resolution transmission electron microscopy and X-ray diffraction measurements were also performed to characterize the samples.

Unexpected roles for bone marrow stromal cells (or MSCs): a real promise for cellular, but not replacement, therapy.

Adult and embryonic stem cells have drawn a lot of attention in the last decade as new tools in regenerative medicine. A variety of such cells have been discovered and put forward as candidates for use in cell replacement therapy. Investigators hope that some, if not all, of our organs can be replaced or restored to function; that new livers, kidneys, and brain cells can be produced. Many reviews have already been written about stem cells and their potential use in regenerating tissues. In this study, we would like to call attention to a different application of a special group of adult stem cells, the stromal cells in the bone marrow (also called mesenchymal stem cells or MSCs). These cells have been discovered to modulate immune function. They can easily be expanded in culture and surprisingly, they also seem not to be immunogenic. Thus, they can be removed from donors, expanded, stored in freezers, and used as allogeneic transplants in a variety of diseases in everyday medicine.

[Significance of arterial stiffness parameters for predicting cardiovascular mortality in hemodialysis patients: a prospective cohort study]. / A különbözô érfali tágulékonysági paraméterek jelentôsége a cardiovascularis mortalitás elôrejelzésében hemodializált betegek között: prospektív kohorszvizsgálat.

UNLABELLED: Previous studies demonstrated that different parameters of arterial stiffness are related to cardiovascular mortality in hemodialysis patients. The relative prognostic value of these parameters has not previously been evaluated in one cohort. PATIENTS AND METHODS: Carotid-femoral pulse wave velocity, carotid augmentation index, carotid pulse pressure and carotid-brachial pulse pressure amplification were measured in 98 patients before and after hemodialysis. Patients were followed for a median of 29 months (1-34) and the association of these parameters with cardiovascular mortality was assessed using log-rank tests and Cox proportional hazards regressions. RESULTS: During follow-up, 40 patients died (mortality rate 20.7/100 patient-year), of which 25 died of cardiovascular causes. Increasing pre- and postdialysis pulse wave velocity tertiles and decreasing predialysis pulse pressure amplification tertiles were significantly related to cardiovascular mortality (p-values are 0.012 and 0.011 for pre- and postdialysis pulse wave velocity, and <0.001 and 0,321 for pre- and postdialysis pulse pressure amplification, respectively). Neither the carotid augmentation index nor carotid pulse pressure was related to cardiovascular mortality. In the Cox-regression, the adjusted hazard ratios for 1 m/s higher pre- and postdialysis pulse wave velocity were 1.24 (1.07-1.44) and 1.17 (1.06-1.28), respectively. The hazard ratio for 10% lower predialysis pulse pressure amplification was 1.41 (1.03-1.92). When included in the same model, both predialysis pulse wave velocity and pulse pressure amplification remained significantly associated with cardiovascular mortality (relative risk: 1.23 [1.07-1.42] and 1.39 [1.02-1.89]). CONCLUSION: Among different stiffness parameters, pulse wave velocity is consistently related to cardiovascular mortality, irrespective of the timing of measurement. Predialysis pulse pressure amplification seems to provide additional prognostic information.