Since the 1970s, about 30 cases of partial or complete trisomy 17p have been presented in the literature. Partial trisomies of the short arm of chromosome 17 are somewhat more common, but complete trisomy is quite rare. Most of these cases were described in infants and newborns; and to our knowledge only 3 cases of trisomy 17p have been detected intrauterine. Phenotypic features of trisomy 17p in fetuses are intrauterine growth retardation, ventriculomegaly, cleft lip and cleft palate, micrognathia, horseshoe kidneys, single umbilical artery, and congenital heart defects. The sonographic and foetopathologic findings of a pregnancy trisomy 17p11.2-pter with the deletion of the terminal portion of the chromosome 6 due to paternal balanced translocation are described in this case report.
Abstract Brown adipocytes, abundant in deep-neck (DN) area in humans, are thermogenic with anti-obesity potential. FTO pro-obesity rs1421085 T-to-C SNP shifts differentiation program towards white adipocytes in subcutaneous fat. Human adipose-derived stromal cells were obtained from subcutaneous neck (SC) and DN fat of 9 donors, of which 3-3 carried risk-free (T/T), heterozygous or obesity-risk (C/C) FTO genotypes. They were differentiated to white and brown (long-term PPARγ stimulation) adipocytes, then global RNA sequencing was performed and differentially expressed genes (DEGs) were compared. DN and SC progenitors had similar adipocyte differentiation potential but differed in DEGs. DN adipocytes displayed higher browning features according to ProFAT or BATLAS scores and characteristic DEG patterns revealing associated pathways which were highly expressed (thermogenesis, interferon, cytokine, retinoic acid, with UCP1 and BMP4 as prominent network stabilizers) or downregulated (particularly extracellular matrix remodelling) compared to SC ones. Part of DEGs in either DN or SC browning was PPARγ-dependent. Presence of the FTO obesity-risk allele suppressed the expression of mitochondrial and thermogenesis genes with a striking resemblance between affected pathways and those appearing in ProFAT and BATLAS, underlining the importance of metabolic and mitochondrial pathways in thermogenesis. Among overlapping regulatory influences which determine browning and thermogenic potential of neck adipocytes, FTO genetic background has a so far not recognized prominence.
Abstract We have previously observed phenotypic and developmental changes upon the ectopic expression of the RUNX3 or the ZBTB46 transcription factors in mouse embryonic stem cell (ESC) derived progenitors. In this study we evaluated the gene expression profiles of the RUNX3- and the ZBTB46-instructed murine ESCs with RNA-Seq testing two next generation sequencing (NGS) technologies. We compared the DNA nanoball (DNB) based MGI DNBSEQ G400 sequencer with the bridge-PCR based Illumina NextSeq 500 instrument. Moreover, we also compared two types of MGI sequencing reagents (Standard- versus Hot-MPS) with the DNBSEQ G400. Importantly, very similar gene expression profile and greatly overlapping RUNX3 and ZBTB46 regulated gene sets were detected with both platforms. Moreover, almost identical gene expression pattern was obtained with the Hot-MPS reagent compared to the Standard-MPS chemistry. This transcriptomic analysis also facilitated the identification of RUNX3 and ZBTB46 regulated genes. For example, we found that Gzmd , Gdf6 and Ccr7 genes were robustly upregulated upon the forced expression of Runx3 , on the other hand, Gpx2 , Tdpoz4 and Arg2 were induced upon the ectopic expression of Zbtb46 . Together these findings demonstrate that the DNBSEQ G400 system is also suitable for global transcript profiling and target gene selection with lower cost.
Abstract We previously described a novel in vitro culture technique for dedifferentiated human adult skin melanocytes. Melanocytes cultured in a defined, cholera toxin and PMA free medium became bipolar, unpigmented, and highly proliferative. Furthermore, TRP‐1 and c‐Kit expression disappeared and EGFR receptor and nestin expression were induced in the cells. Here, we further characterized the phenotype of these dedifferentiated cells and by comparing them to mature pigmented melanocytes we detected crucial steps in their phenotype change. Our data suggest that normal adult melanocytes easily dedifferentiate into pluripotent stem cells given the right environment. This dedifferentiation process described here for normal melanocyte is very similar to what has been described for melanoma cells, indicating that phenotype switching driven by environmental factors is a general characteristic of melanocytes that can occur independent of malignant transformation.
Background: In sepsis, platelet activation by lipopolysaccharide (LPS) via TLR4 can result in microvascular thrombosis. However, megakaryocytes (MKs) also express TLRs, thus severe infection can modulate thrombopoiesis. Both MKs and platelets are rich in microRNAs (miRNAs) to regulate messenger RNA (mRNA) function and different cellular events. Murine MKs were previously described to produce platelets with altered mRNA profile in septic mice. Aims: We characterized sepsis‐induced expression of mRNAs with their regulatory miRNAs in ex vivo and in vitro stimulated platelets as well as in MK cell cultures. The contribution of Dicer function to abnormal miRNA levels was also investigated. Methods: Leukocyte‐depleted platelets of 22 septic and 26 healthy individuals were analyzed for P2RY12 and SELP (P‐selectin) mRNAs. TaqMan Open Array was performed for miRNAs, and RT‐qPCR were used for verification. The effects of sepsis were also studied in isolated platelets and MEG‐01 cells in vitro after treatment with recombinant TNF‐α (100 ng/mL) or LPS (O55:B5, 100 ng/mL) with lipoprotein binding protein (100 ng/mL) and soluble CD14 (150 ng/mL) for 1–24 h. Dicer level was observed by western blotting and fluorescence microscope, while its function was tested by calpain inhibitor (calpeptin, 10 μmol/L) in septic MKs and silencing by Dicer siRNA (40 pmol) in MEG‐01 cells in comparison to control samples with NEG‐01 siRNA for P2RY12 and SELP expression. To prove the effect of LPS, TLR4 expression was observed on platelets and MKs by flow cytometry. Induced inflammatory conditions were evaluated by miR‐155 expression and IL‐1β/IL‐6 mRNA levels by RT‐qPCR and via nuclear translocation of p65 subunit of NF‐κB pathway by fluorescence microscope. Results: There was augmented platelet activation as surface and soluble P‐selectin were increased in septic patients ( P < 0.001). Platelets and MKs upregulated P2RY12 and SELP gene expression ( P < 0.01) accompanied with elevated inflammation‐specific miR‐155 and decreased miR‐223/26b. Additionally, 64 other platelet miRNAs (e.g. miR‐150, let‐7e) indicated ≥2‐fold decrease, while 37 (e.g. miR‐191) were increased at the same degree in sepsis vs. controls. Septic conditions resulted in substantial p65 translocation into nuclei causing at least 2‐fold elevated miR‐155 and IL‐1β or IL‐6 mRNA levels ( P < 0.05) in platelets and MKs in the ex vivo and in vitro experiments via TLR4 receptors that could be detected on the surface of platelets (3–7%) and MEG‐01 cells (8–12%). After a transient induction of Dicer and miRNAs at 1 h, reduced Dicer mRNA and protein levels were seen in LPS/TNF‐α treated platelets/MKs after 4–24 h causing attenuated miR‐223/26b. Calpain inhibition restored miRNA levels, while downregulation of Dicer generated higher P2RY12 and SELP levels in MEG‐01. Summary/Conclusion: Alteration in Dicer‐dependent miRNAs contribute to elevated mRNA levels in MKs and reactive platelets during sepsis. This study is supported by the GINOP‐2.3.2–15–2016–00043‐IRONHEART project.
Acne vulgaris provides a unique disease setting in which a prominent skin inflammation is coupled with the overproduction of lipid-rich sebum.Our goal was to evaluate the expression of barrier molecules in papular acne skin samples obtained from untreated patients and compare those to the results of healthy and of papulopustular rosacea-involved ones at the mRNA and protein levels. In addition, we aimed to assess the effects of various sebum composing lipids on the expression of proteins involved in barrier formation in keratinocytes.Available microarray data sets of papular acne and papulopustular rosacea-affected skin samples were re-analysed with a focus on epidermal barrier-related pathways. Immunohistochemistry was performed to detect barrier molecules in the interfollicular regions of human acne and healthy skin samples. Protein levels of barrier-related genes were measured by western blot in samples of HaCaT keratinocytes treated with selected lipids.Meta-analysis of whole transcriptome data sets revealed that barrier-related pathways are significantly affected in acne vulgaris skin samples. While an altered expression of key molecules in maintaining barrier functions such as filaggrin, keratin 1, involucrin, desmoglein 1, kallikrein 5 and 7, was also observed at the protein levels, our data demonstrated that sebum composing lipids may selectively modify the levels of epidermal barrier-related molecules.Our results suggest that although not as prominently as in the dry papulopustular rosacea skin, the epidermal barrier in the interfollicular region may be damaged also in the lipid-rich skin samples of papular acne. Furthermore, our findings indicating diverse regulatory effects of various sebum lipids on the expression of barrier molecules in keratinocytes suggest, that they may influence the moisturization of the skin as well. Altogether, our findings could have implications in the development of sebum-modulating anti-acne therapies and even in the care of symptom-free skin.
