Abstract 167: Mir-195-3p/-5p Decrease Cardiac Fibroblast Proliferation and the Transdifferentiation into Myofibroblasts

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Shutong Shen ◽  
Xiuzhi Wang ◽  
Dongjie Xu ◽  
Lichan Tao ◽  
Xiaoting Wu ◽  
...  
Keyword(s):  
Target Gene ◽  
Cardiac Fibrosis ◽  
Cardiac Fibroblast ◽  
Immunofluorescent Staining ◽  
Fibroblast Proliferation ◽  
Chain Reactions ◽  
Protein Levels ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain

Aims: MicroRNAs (miRNAs, miRs) contribute to many essential physiological and pathological processes including fibrosis. This study aims at investigating the role of miR-195-3p/-5p in cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts. Methods and results: In isolated primary neonatal cardiac fibrobasts (NRCFs), forced expression of miR-195-3p/-5p with agomiRs could attenuate fibrobast proliferation as determined by EdU and Ki67 staining while inhibition of miR-195-3p/-5p with antagomiRs could increase fibrobast proliferation. By quantitative reverse transcription polymerase chain reactions (RT-PCRs) and western blotting (WB), α-SMA (a marker of myofibroblast transdifferentiation) was found to be suppressed in the miR-195-3p/-5p agomiR-treated NRCFs at both mRNA and protein levels, while was increased in the miR-195-3p/-5p antagomiR-treated NRCFs. Moreover, Chek-1 was identified as a target gene of miR-195-3p/-5p responsible for cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts by RT-PCR and WB and immunofluorescent staining. Silencing of Chek-1 attenuates cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts as detected by α-SMA/EDU staining. In addition, Chek-1 mediated the effects of miR-195-3p/-5p in cardiac fibroblast proliferation and the transdifferentiation into myofibroblasts. Conclusion: Therefore, miR-195-3p/-5p might be promising therapeutic targets for cardiac fibrosis.

Molecular evidence of vector-borne pathogens coinfecting dogs from Poland

2011 ◽  
Vol 59 (2) ◽  
pp. 215-223 ◽  
Author(s):  
Anna Rymaszewska ◽  
Małgorzata Adamska
Keyword(s):  
Anaplasma Phagocytophilum ◽  
Molecular Evidence ◽  
Chain Reactions ◽  
Group Iii ◽  
Polymerase Chain Reactions ◽  
Group I ◽  
Vector Borne ◽  
Polymerase Chain ◽  
Pathogenic Agents

Ticks of the genusIxodesare vectors for many pathogens, includingBorrelia burgdorferisensu lato,Anaplasma phagocytophilumandRickettsiaspp., and may also serve as vectors forBartonellaspp. However, the role of ticks inBartonellatransmission requires additional studies. The aim of this study was to investigate whether coinfection with two or more vector-borne pathogens can occur in the following three groups of dogs: I — dogs with suspected borreliosis (N = 92), II — dogs considered healthy (N = 100), and III — dogs with diagnosed babesiosis (N = 50). Polymerase chain reactions were performed to detect DNA ofAnaplasma phagocytophilum, Rickettsiaspp. andBartonellaspp. in the blood of dogs. In dogs of Group I, the DNA of bothA. phagocytophilumandBartonellasp. was detected (14% and 1%, respectively). In eight dogs, coinfection was indicated:A. phagocytophilumorBartonellasp. withB. burgdorferis.l. (the presence of antibodies against and/or DNAB. burgdorferis.l.). In the case of five dogs positive forA. phagocytophilumDNA, no coinfection withB. burgdorferis.l. was shown. In Group II, the DNA ofA. phagocytophilumwas detected in four dogs. In Group III, no pathogenic agents possibly transmitted by ticks were confirmed. No DNA ofR. helveticawas detected in any of the groups studied.

scholarly journals High number of asymptomatic dogs as leptospiral carriers in an endemic area indicates a serious public health concern

2017 ◽  
Vol 145 (9) ◽  
pp. 1852-1854 ◽  
Author(s):  
R. SANT'ANNA ◽  
A. S. VIEIRA ◽  
J. GRAPIGLIA ◽  
W. LILENBAUM
Keyword(s):  
Public Health ◽  
Endemic Area ◽  
Urine Samples ◽  
Health Concern ◽  
Endemic Region ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Knowing That ◽  
Polymerase Chain

SUMMARYAsymptomatic dogs can be potential hosts of leptospirosis. However, the extension of this phenomenon in endemic areas has not yet been clearly defined. This study is aimed at evaluating the role of asymptomatic dogs as carriers of Leptospira in an endemic area of Brazil. A total of 131 male dogs without apparent leptospirosis symptoms were included in the study based on clinical and hematologic exams. Serum and urine samples were collected for microscopic agglutination tests (MAT) and polymerase chain reactions (PCR) targeted the LipL32 gene, respectively. Forty-two dogs (32·1%) presented seroreactivity (titres ⩾100). The serogroup Icterohaemorrhagiae was predominant, representing 92·7% of the seropositive samples. Overall, leptospiral DNA was detected on 26 urine samples (19·8%). PCR positivity was more common (28·6%) on seropositive dogs than on seronegative (15·7%) ones. Nevertheless, MAT was not correlated to PCR (P > 0·05). Age was not associated with seroreactivity, but dogs older than 5 years of age had 4·07 more chances (odds ratio) of being carriers (PCR positive) than younger ones. Although the fact of knowing that asymptomatic dogs can act as leptospiral carriers is not new, the extension of this fact is impressive in an endemic region, and its role and impact on public health cannot be neglected.

Identification of the NO Synthase isoforms Expressed in Human Neutrophil Granulocytes, Megakaryocytes and Platelets

1997 ◽  
Vol 77 (01) ◽  
pp. 163-167 ◽  
Author(s):  
Thomas Wallerath ◽  
Ingolf Gath ◽  
Walter E Aulitzky ◽  
Jennifer S Pollock ◽  
Hartmut Kleinert ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Human Platelets ◽  
No Synthase ◽  
Immunofluorescent Staining ◽  
Human Neutrophils ◽  
Neutrophil Granulocytes ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain

SummaryUsing Western blot and fluorescent immunocytochemistry, NOS III (or ecNOS) and NOS II (or iNOS), but no NOS I (or ncNOS), were identified in preparations of human platelets. Reverse-transcription polymerase chain reactions (RT-PCR) demonstrated NOS III mRNA, but no NOS II mRNA (which is short-lived) and no NOS I mRNA in platelets. Immunofluorescent staining of human bone marrow smears showed the presence of NOS III, but not NOS I in megakaryocytes. A subpopulation of megakaryocytes also expressed NOS II. In preparations of human neutrophils, immunocytochemistry demonstrated NOS I in all cells, whereas no NOS III was detected. The few NOS II positive cells were characterized as contaminating eosinophils. Similarly, in RT-PCR, transcripts for NOS I and NOS II, but not for NOS III, were identified. Thus, the constitutive NOS isoform in megakaryocytes and platelets is NOS III, whereas neutrophils express NOS I. Some megakaryocytes and eosinophils also express NOS II.

scholarly journals Kinship in Aegean Prehistory? Ancient DNA in Human Bones from Mainland Greece and Crete

2009 ◽  
Vol 104 ◽  
pp. 293-309 ◽  
Author(s):  
Abigail S. Bouwman ◽  
Keri A. Brown ◽  
Terence A. Brown ◽  
Elizabeth R. Chilvers ◽  
Robert Arnott ◽  
...  
Keyword(s):  
Bronze Age ◽  
Social Relationships ◽  
Ancient Dna ◽  
Eastern Mediterranean ◽  
High Status ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain ◽  
The Bronze Age

Attempts were made to detect ancient DNA (aDNA) in samples of 89 human skeletons from Neolithic and Bronze Age sites in Greece and Crete. Ancient DNA was absent in specimens from Nea Nicomedia, Lerna, Kato Zakro: Karaviádena, and Mycenae Grave Circle A. For each of three skeletons sampled from Antron Grave Circle B, polymerase chain reactions (PCRs) gave products for nuclear but not mitochondrial DNA, but the yield of DNA was low and inconsistent, with replicate PCRs failing to give reproducible results. At Kouphovouno evidence for mitochondrial and/or nuclear aDNA was obtained from eight of the 20 skeletons that were examined, while at Mycenae Grave Circle B evidence for mitochondrial aDNA was obtained for four of the 22 skeletons that were studied, and in two cases confirmed the evidence of close kinship that had already been suggested by facial reconstruction: this in turn raises interesting questions of social relationships and the role of high-status women in MBA/LBA society. We conclude that, although aDNA might be present in some Eastern Mediterranean skeletons from later centuries of the Bronze Age, it is not commonly found in material from this period and is likely to be absent from older material.Στη μελέτη αυτή έγιναν προσπάθειες να αναγνωριστεί αρχαίο DNA (aDNA) σε δείγματα ογδόντα εννέα ανθρώπινων σκελετών προερχομένων από θέσεις της Νεολιθικής περιόδου και της Εποχής του Χαλκού στην Ελλάδα και την Κρήτη. Αρχαίο DNA δεν εντοπίστηκε σε δείγματα από τη Νέα Νικομήδεια, τη Λέρνα, την Κάτω Ζάκρο (Καραβιάδενα) και τον Ταφικό Κύκλο Α των Μυκηνών. Για κάθε έναν από τους τρεις σκελετούς, οι οποίοι εξετάστηκαν από τον Ταφικό Κύκλο Β της Αντρώνας, οι αλυσιδωτές αντιδράσεις πολυμεράσης (PCRs) απέφεραν αποτελέσματα για πυρηνικό αλλά όχι μιτοχονδριακό DNA. Η παραγωγή DNA ήταν χαμηλή και αντιφατική, με τα αντίγραφα πολυμεράσης να αποτυγχάνουν να αποφέρουν αναπαραγώγιμα αποτελέσματα. Στο Κουφόβουνο οκτώ από τους είκοσι σκελετούς, που εξετάστηκαν, έδωσαν στοιχεία για μιτοχονδρνακό ή/και πυρηνικό DNA, ενώ στον Ταφικό Κύκλο Β των Μυκηνών ενδείξεις για μιτοχονδριακό DNA έδωσαν τέσσερεις από τους είκοσι δύο σκελετούς, που μελετήθηκαν. Σε δύο περιπτώσεις επιβεβαιώθηκε η ένδειξη στενής συγγένειας, κάτι το οποίο είχε ήδη προταθεί με την αποκατάσταση των προσώπων: το γεγονός αυτό εγείρει ενδιαφέροντα ερωτήματα σχετικά με τις κοινωνικές σχέσεις και το ρόλο γυναικών υψηλής κοινωνικής στάθμης στην κοινωνία της Μέσης και της Ύστερης Εποχής του Χαλκού. Συμπεραίνουμε ότι, αν και μπορεί να αναγνωριστεί DNA σε ορισμένους σκελετούς της Ανατολικής Μεσογείου των τελευταίων αιώνων της Εποχής του Χαλκού, δεν εντοπίζεται συχνά σε υλικό αυτής της εποχής και ενδεχομένως απουσιάζει από παλαιότερο υλνκό.

scholarly journals Atg16L1 as a Novel Biomarker and Autophagy Gene for Diabetic Retinopathy

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Xinxiao Gao ◽  
Yunhui Du ◽  
Wayne Bond Lau ◽  
Yu Li ◽  
Siquan Zhu ◽  
...  
Keyword(s):  
Diabetic Retinopathy ◽  
Real Time ◽  
Potential Role ◽  
Critical Role ◽  
Differentially Expressed ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain ◽  
Microarray Analyses

Objective. Accumulating evidence suggests the critical role of autophagy in the pathogenesis of diabetic retinopathy (DR). In the current study, we aim to identify autophagy genes involved in DR via microarray analyses. Methods. Gene microarrays were performed to identify differentially expressed lncRNAs/mRNAs between normal and DR retinas. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses of lncRNA-coexpressed mRNAs were used to determine the related pathological pathways and biological modules. Real-time polymerase chain reactions (PCR) were conducted to validate the microarray analyses. Results. A total of 2474 significantly dysregulated lncRNAs and 959 differentially expressed mRNAs were identified in the retina of DR. Based upon Signalnet analysis, Bcl2, Gabarapl2, Atg4c, and Atg16L1 participated the process of cell death in DR. Moreover, real-time PCR revealed significant upregulation of Atg16L1. Conclusion. This study indicated the importance and potential role of Atg16L1, one of the autophagy genes, as a biomarker in DR development and progression.

scholarly journals The two faces of mast cells in vitiligo pathogenesis

2021 ◽  
Author(s):  
Ichiro Katayama ◽  
Lingli Yang ◽  
Aya Takahashi ◽  
Fei Yang ◽  
Mari Wataya-Kaneda
Keyword(s):  
Mast Cells ◽  
Immunofluorescent Staining ◽  
Epidermal Keratinocytes ◽  
T Helper ◽  
T Helper 17 ◽  
Double Positive ◽  
Protein Levels ◽  
Polymerase Chain

Aim: Previously, we reported increased number of T helper 17 (Th17) cells in vitiligo. However, in our recent study, tryptase and interleukin (IL)17 double positive cells which identified by polyclonal anti-IL17 antibody with specificity for IL17A, B, D, F was observed, but these mast cells cannot be stained by monoclonal anti-IL17 antibody with specificity for IL17A. Therefore, this study was aimed to clarify the role of mast cells in induction and progression of vitiligo. Methods: Mast cells were stained with two antibodies against IL17 and one antibody against tryptase by immunofluorescent staining. Furthermore, immunoelectron microscopy (IEM) analyses were conducted using anti-tryptase. In vitro, cultured epidermal keratinocytes were treated with agents which released by mast cells. Expression levels of mRNA were analyzed by real-time polymerase chain reaction (PCR), expression of protein levels was analyzed by western blotting. Results: An increased number of tryptase positive mast cells was observed at the lesional skin of upper dermis in vitiligo and rhododendrol-induced leukoderma (RDIL). These mast cells showed prominent degranulation in vitiligo. Interestingly, the melanosome forming glycoprotein non-metastatic melanoma protein B (GPNMB) is downregulated in the lesional basal keratinocytes in vitiligo and mast cell tryptase contributes to this phenomenon. In addition, small interfering GPNMB RNA (siGPNMB RNA)-introduced keratinocytes increased melanocyte survival through stem cell factor (SCF) production in the melanocyte/keratinocyte co-culture system. Conclusions: Mast cells might be two-faced in vitiligo induction, progression, and recovery through the differential function of histamine and tryptase.

scholarly journals Single and Mixed Phytoplasma Infections in Phyllody- and Dwarf-Diseased Clover Plants in Lithuania

Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1061-1066 ◽  
Author(s):  
Juozas B. Staniulis ◽  
Robert E. Davis ◽  
Rasa Jomantiene ◽  
Audrone Kalvelyte ◽  
Ellen L. Dally
Keyword(s):  
16S Rdna ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Restriction Sites ◽  
Polymerase Chain ◽  
Dwarf Disease ◽  
Clover Phyllody

Naturally diseased plants of clover (Trifolium spp.) exhibiting symptoms of clover phyllody (virescence and phyllody of flowers) or of clover dwarf (abnormally small leaves, shortened internodes, proliferation of shoots, and dwarf growth habit) were observed in fields in Lithuania. Phytoplasma group-specific polymerase chain reactions (PCRs) and restriction fragment length polymorphism (RFLP) analysis of 16S rDNA revealed that the plants were infected by two mutually distinct phytoplasmas. Clover phyllody-diseased plants were infected by a subgroup 16SrI-C (subgroup I-C) phytoplasma (CPh-L) related to clover phyllody (CPh-C) phytoplasma in Canada. Clover dwarf-diseased plants were infected by both CPh-L and a phytoplasma (CYE-L) related to clover yellow edge (CYE-C) phytoplasma (subgroup 16SrIII-B = III-B) in Canada. A 1.8-kbp fragment of rRNA operon from CYE-L was amplified, cloned, and sequenced, and putative restriction sites mapped. This sequence shared high similarity (99.7%) with that of CYE-C and exhibited no differences from CYE-C in RFLP patterns of 16S rDNA; therefore, we tentatively classified CYE-L in subgroup 16SrIII-B (type strain, CYE = CYE-C phytoplasma) of the X-disease phytoplasma group. These findings extend the known geographical ranges of subgroup I-C and subgroup III-B taxa to the region of northern Europe including Lithuania and suggest a role of the subgroup III-B phytoplasma in clover dwarf disease.

Sign in / Sign up

Export Citation Format

Share Document