scholarly journals ARG1 could be expressed by human, but not represent for repair

2020 ◽  
Author(s):  
Chenyu Chu ◽  
Chen Hu ◽  
Shengan Rung ◽  
Yufei Wang ◽  
Yili Qu ◽  
...  
Keyword(s):  
Bone Tissue ◽  
Quantitative Polymerase Chain Reaction ◽  
Healing Process ◽  
Tissue Samples ◽  
Protein Levels ◽  
Arginase 1 ◽  
Tissue Healing ◽  
Human Bone Tissue ◽  
Polymerase Chain ◽  
Clinical Healing

AbstractArginase 1 (ARG1), typically expressed under stress or stimulation rather than in the healthy condition in humans. However, more and more studies claim ARG1 could be expressed by human and represents healing process. Including THP-1 monocyte shift to M2 macrophages rendered a higher expression of ARG1 than that with bone-graft has an ideal effect on anti-inflammation, resulting in tissue healing. With the higher expression of ARG1 seemingly induced by the constructed material and represented by the anti-inflammatory situation. For these reasons, this study focuses on whether traditional markers of macrophages could be express by human or not during clinical healing process. Through harvesting bone tissue samples during bone healing process and confirm the expression of markers at transcriptome and protein levels. The results showed the expression of ARG1 relatively low by bulk-RNA sequencing, quantitative polymerase chain reaction (qPCR), immunofluorescence (IF) and flow cytometry or could not be detected during process of healing of healthy donor. Taken together, results indicate that the ARG1 is not chosen for a regenerative marker in human bone tissue.

scholarly journals Active liver X receptor signaling in phagocytes in multiple sclerosis lesions

2017 ◽  
Vol 24 (3) ◽  
pp. 279-289 ◽  
Author(s):  
Jo Mailleux ◽  
Tim Vanmierlo ◽  
Jeroen FJ Bogie ◽  
Elien Wouters ◽  
Dieter Lütjohann ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Monocyte ◽  
Liver X Receptor ◽  
Spectrometric Analysis ◽  
Protein Levels ◽  
Myelin Phagocytosis ◽  
Polymerase Chain ◽  
Cholesterol Precursors ◽  
Chromatographic Mass

Objective: We sought to determine the liver X receptor (LXR) ligands present in human macrophages after myelin phagocytosis and whether LXRs are activated in multiple sclerosis (MS) lesions. Methods: We used real-time quantitative polymerase chain reaction (PCR) and immunohistochemistry to determine expression of LXRs and their response genes in human phagocytes after myelin phagocytosis and in active MS lesions. We used gas chromatographic/mass spectrometric analysis to determine LXR-activating oxysterols and cholesterol precursors present and formed in myelin and myelin-incubated cells, respectively. Results: Myelin induced LXR response genes ABCA1 and ABCG1 in human monocyte-derived macrophages. In active MS lesions, we found that both gene expression and protein levels of ABCA1 and apolipoprotein E ( APOE) are upregulated in foamy phagocytes. Moreover, we found that the LXR ligand 27-hydroxycholesterol (27OHC) is significantly increased in human monocyte-derived macrophages after myelin uptake. Conclusion: LXR response genes are upregulated in phagocytes present in active MS lesions, indicating that LXRs are activated in actively demyelinating phagocytes. In addition, we have shown that myelin contains LXR ligands and that 27OHC is generated in human monocyte-derived macrophages after myelin processing. This suggests that LXRs in phagocytes in active MS lesions are activated at least partially by (oxy)sterols present in myelin and the generation thereof during myelin processing.

scholarly journals Decreased Expression of GPER1 Gene and Protein in Goiter

2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Raquel Weber ◽  
Ana Paula Santin Bertoni ◽  
Laura Walter Bessestil ◽  
Ilma Simoni Brum ◽  
Tania Weber Furlanetto
Keyword(s):  
Estrogen Receptor ◽  
Quantitative Polymerase Chain Reaction ◽  
Thyroid Tissue ◽  
Chain Reaction ◽  
Normal Thyroid ◽  
Protein Levels ◽  
Protein Expressions ◽  
Polymerase Chain ◽  
G Protein Coupled

Goiter is more common in women, suggesting that estrogen could be involved in its physiopathology. The presence of classical estrogen receptors (ERαand ERβ) has been described in thyroid tissue, suggesting a direct effect of estrogen on the gland. A nonclassic estrogen receptor, the G-protein-coupled estrogen receptor (GPER1), has been described recently in several tissues. However, in goiter, the presence of this receptor has not been studied yet. We investigated GPER1 gene and protein expressions in normal thyroid and goiter using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot, respectively. In normal thyroid (n=16) and goiter (n=19), GPER1 gene was expressed in all samples, while GPER1 protein was expressed in all samples of normal thyroid (n=15) but in only 72% of goiter samples (n=13). When comparing GPER1 gene and protein levels in both conditions, gene expression and protein levels were higher in normal thyroid than in goiter, suggesting a role of this receptor in this condition. Further studies are needed to elucidate the role of GPER1 in normal thyroid and goiter.

scholarly journals Total Versus Free Placental Growth Factor Levels in the Pathogenesis of Preeclampsia

Hypertension ◽  
2020 ◽  
Vol 76 (3) ◽  
pp. 875-883
Author(s):  
Edouard Lecarpentier ◽  
Zsuzsanna K. Zsengellér ◽  
Saira Salahuddin ◽  
Ambart E. Covarrubias ◽  
Agnes Lo ◽  
...  
Keyword(s):  
Growth Factor ◽  
Ex Vivo ◽  
Quantitative Polymerase Chain Reaction ◽  
Placental Growth Factor ◽  
Protein Levels ◽  
Key Characteristics ◽  
Human Blood Samples ◽  
Polymerase Chain ◽  
Low Levels ◽  
Independent Cohort

Elevated circulating sFLT-1 (soluble fms-like tyrosine kinase) and low levels of its ligand, PlGF (placental growth factor), are key characteristics of preeclampsia. However, it is unclear if the low levels of plasma PlGF noted during preeclampsia are due to decreased placental production of PlGF or due to binding of PlGF by increased circulating sFLT-1. Here, we describe a biochemical procedure to dissociate PlGF-sFLT-1 complex ex vivo and when used in conjunction with an immunoassay platform, demonstrate a method to measure total and free PlGF in human blood samples. Using this method, we noted that plasma free PlGF levels were significantly lower in preeclampsia (N=22) than in nonhypertensive controls (N=24; mean, 314 versus 686 pg/mL, P <0.05), but total PlGF levels were not different (mean, 822 versus 800 pg/mL, P =0.49). In contrast, total sFLT-1 levels were significantly higher in preeclampsia than in nonhypertensive controls (mean, 16 957 versus 3029 pg/mL, P <0.01) and sFLT-1 levels correlated with bound PlGF levels (bound PlGF=total PlGF−free PlGF) in these samples ( r 2 =0.68). We confirmed these findings in an independent cohort of subjects (N=49). Furthermore, we did not detect any difference in PlGF mRNA by quantitative polymerase chain reaction or in PlGF protein expression by immunohistochemistry in preeclamptic placentas when compared with nonhypertensive controls. In contrast, sFLT-1 mRNA and protein levels were upregulated in placentas from women with preeclampsia. Taken together with prior studies, our results provide evidence that decrease in circulating PlGF noted during preeclampsia is largely mediated by excess circulating sFLT-1.

scholarly journals OX40+ T lymphocytes and IFN-γ are associated with American tegumentary leishmaniasis pathogenesis

2012 ◽  
Vol 87 (6) ◽  
pp. 851-855
Author(s):  
Patrícia Luciana Batista Domingos ◽  
Agostinho Gonçalves Viana ◽  
Carlos Alberto de Carvalho Fraga ◽  
Paulo Rogério Ferreti Bonan
Keyword(s):  
Immune Response ◽  
Cutaneous Leishmaniasis ◽  
Healing Process ◽  
Public Health Problem ◽  
Leishmania Species ◽  
Tissue Samples ◽  
Protein Levels ◽  
Skin Ulcers ◽  
American Tegumentary Leishmaniasis ◽  
Ifn Γ

BACKGROUND: Leishmaniases are zoonoses considered a public health problem, representing a complex group of diseases with a broad clinical spectrum and epidemiological diversity. Leishmaniasis is caused by several species of protozoa of the genus Leishmania. The evolution of the pathology and the resolution of the leishmaniasis are dependent mainly on the Leishmania species involved, although the cytokine profile plays an important role in the development of the immune response. OBJECTIVES: The purpose of our study was to evaluate the immune response of patients affected by lesions of cutaneous leishmaniasis by immunostaining of the OX40, CD20, IFN-γ and IL-4 proteins. METHODS: The tissue samples were collected from indolent skin ulcers confirmed as cutaneous leishmaniasis of 41 patients aged between six and 90 years. The lesions were submitted to OX40, CD20, INF-γ and IL-4 immunolabeling. RESULTS: We observed a statistically significant higher expression of IFN-γ compared with IL-4 (p=0.009). Besides, OX40 had higher expression when compared with CD20 (p<0.001). CONCLUSION: The present study indicates that the immune response in lesions of cutaneous leishmaniasis is associated with a healing process, which can be explained by the higher expression of IFN-γ when compared with IL4 protein levels.

A comprehensive search for microbial DNA and RNA in sarcoidosis tissue samples by quantitative polymerase chain reaction

2017 ◽  
Vol 49 (8) ◽  
pp. 625-627
Author(s):  
Fumio Terasaki ◽  
Hitomi Fukumoto ◽  
Ryo Kawata ◽  
Yoshinobu Hirose ◽  
Shu-ichi Fujita ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Dna And Rna ◽  
Comprehensive Search ◽  
Polymerase Chain ◽  
Microbial Dna

scholarly journals Gene Expression Changes in Femoral Head Necrosis of Human Bone Tissue

2011 ◽  
Vol 31 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Bernadett Balla ◽  
Csaba Pintér ◽  
János P. Kósa ◽  
János Podani ◽  
István Takács ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Femoral Head ◽  
Bone Tissue ◽  
Expression Profile ◽  
Human Bone ◽  
Taqman Probe ◽  
Local Circulation ◽  
Tissue Samples ◽  
Human Bone Tissue

Osteonecrosis of the femoral head (ONFH) is the result of an interruption of the local circulation and the injury of vascular supply of bone. Multiple factors have been implicated in the development of the disease. However the mechanism of ischemia and necrosis in non-traumatic ONFH is not clear. The aim of our investigation was to identify genes that are differently expressed in ONFH vs. non-ONFH human bone and to describe the relationships between these genes using multivariate data analysis. Six bone tissue samples from ONFH male patients and 8 bone tissue samples from non-ONFH men were examined. The expression differences of selected 117 genes were analyzed by TaqMan probe-based quantitative real-time RT-PCR system. The significance test indicated marked differences in the expression of nine genes between ONFH and non-ONFH individuals. These altered genes code for collagen molecules, an extracellular matrix digesting metalloproteinase, a transcription factor, an adhesion molecule, and a growth factor. Canonical variates analysis demonstrated that ONFH and non-ONFH bone tissues can be distinguished by the multiple expression profile analysis of numerous genes controlled via canonical TGFB pathway as well as genes coding for extracellular matrix composing collagen type molecules. The markedly altered gene expression profile observed in the ONFH of human bone tissue may provide further insight into the pathogenetic process of osteonecrotic degeneration of bone.

scholarly journals Effects of active acromegaly on bone mRNA and microRNA expression patterns

2018 ◽  
Vol 178 (4) ◽  
pp. 353-364 ◽  
Author(s):  
Zhanna Belaya ◽  
Tatiana Grebennikova ◽  
Galina Melnichenko ◽  
Alexey Nikitin ◽  
Alexander Solodovnikov ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Bone Tissue ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Sella Turcica ◽  
Compensatory Mechanisms ◽  
Tissue Samples ◽  
Active Acromegaly ◽  
Cell Commitment

Objective To evaluate the response of bone to chronic long-term growth hormone (GH) and insulin-like growth factor-1 (IGF1) excess by measuring the expression of selected mRNA and microRNA (miR) in bone tissue samples of patients with active acromegaly. Design Case–control study. Methods Bone tissue samples were obtained during transsphenoidal adenomectomy from the sphenoid bone (sella turcica) from 14 patients with clinically and biochemically confirmed acromegaly and 10 patients with clinically non-functioning pituitary adenoma (NFPA) matched by sex and age. Expression of genes involved in the regulation of bone remodeling was studied using quantitative polymerase chain reaction (qPCR). Results Of the genes involved in osteoblast and osteoclast activity, only alkaline phosphatase (ALP) mRNA was 50% downregulated in patients with acromegaly. GH excess caused increased expression of the Wnt signaling antagonists (DKK1) and agonists (WNT10B) and changes in the levels of miR involved in mesenchymal stem cell commitment to chondrocytes (miR-199a-5p) or adipocytes (miR-27-5p, miR-125b-5p, miR-34a-5p, miR-188-3p) P < 0.05; q < 0.1. Relevant compensatory mechanisms were found through the changes in miR involved in osteoblastogenesis (miR-210-5p, miR-135a-5p, miR-211, miR-23a-3p, miR-204-5p), but the expression of TWIST1 was 50% downregulated and RUNX2 was unchanged. Conclusions Acromegaly had minimal effects on tested mRNAs specific to osteoblast or osteoclast function except for downregulated ALP expression. The expressions of miR known to be involved in mesenchymal stem cell commitment and downregulated TWIST1 expression suggest acromegaly has a negative effect on osteoblastogenesis.

Expression and cellular distribution of zona pellucida glycoproteins in canine oocytes before and after in vitro maturation

Zygote ◽  
2014 ◽  
Vol 23 (6) ◽  
pp. 863-873 ◽  
Author(s):  
Bartosz Kempisty ◽  
Hanna Piotrowska ◽  
Dorota Bukowska ◽  
Magdalena Woźna ◽  
Sylwia Ciesiółka ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Quantitative Polymerase Chain Reaction ◽  
Microscopic Analysis ◽  
Maturation Stage ◽  
Protein Distribution ◽  
Protein Levels ◽  
Peripheral Area ◽  
Before And After ◽  
Polymerase Chain

SummaryThis study was aimed at investigating zona pellucida glycoproteins (ZP) ZP2, ZP3 mRNA expression as well as ZP3, ZP4 (ZPB) protein distribution before and after in vitro maturation (IVM) in canine oocytes. The cumulus–oocyte complexes (COCs) were recovered from 27 anoestrous mongrel bitches and matured for 72 h in TCM199 medium. The canine COCs were analysed before and after IVM. Using real-time quantitative polymerase chain reaction (RQ-PCR), both groups of oocytes were analysed for detection of ZP2 and ZP3 mRNA profiles as well as using confocal microscopic analysis for observation of ZP3 and ZP4 protein distribution. In post-IVM canine oocytes an increase in transcript content of ZP2 and ZP3 genes as well as a decrease in ZP3 and ZP4 protein levels were observed when compared with pre-IVM oocytes. Moreover, the ZP4 protein before IVM was significantly distributed in the peripheral area of cytoplasm, whereas after IVM it was localized rather than in the entire cytoplasm. In contrast, the ZP3 protein was found both before and after IVM was distributed in the peripheral area of the cytoplasm. In conclusion, we suggest that the expression of ZP2 and ZP3 genes is associated with the maturation stage of canine oocytes, as higher mRNAs levels were found after IVM. However, a decreased expression of ZP3 and ZP4 proteins after IVM suggests maturation-dependent down-regulation of these protein translations, which may result in disturbed fertilization.

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