Cytokine and Growth Factor mRNA Expression Patterns Associated with the Hypercontracted, Hyperpigmented Healing Phenotype of Red Duroc Pigs: A Model of Abnormal Human Scar Development?

2005 ◽  
Vol 9 (4) ◽  
pp. 165-177 ◽  
Author(s):  
Corrie L. Gallant—Behm ◽  
Merle E. Olson ◽  
David A. Hart
Keyword(s):  
Mrna Expression ◽  
Expression Patterns ◽  
Hypertrophic Scars ◽  
Rt Pcr ◽  
Specific Primers ◽  
Juvenile Female ◽  
Biphasic Pattern ◽  
Porcine Gene ◽  
Molecular Expression ◽  
Rna And Dna

Background: Skin wounds in red Duroc pigs heal with the formation of hypercontractile, hyperpigmented scars, similar in some respects to human hypertrophic scars. ObjectiveThe goal of this study was to characterize the mRNA expression patterns for a subset of relevant cytokines, growth factors, receptors, and transcription factors involved in the red Duroc scarring phenotype. Methods: Full-thickness and deep dermal wounds were created on the backs of juvenile female red Duroc pigs. Samples were taken every two weeks postwounding and total RNA and DNA were extracted and quantified. RT-PCR was performed using porcine gene-specific primers for 15 relevant molecules. Results: The majority of molecules examined exhibited a biphasic pattern of expression, with peaks of expression at days 14 and 56 postinjury. Conclusions: The molecular expression pattern observed correlates well with the gross healing phenotype and matrix molecule expression patterns previously reported in red Duroc pigs. These findings enhance our understanding of the processes associated with fibroproliferative scar-formation.

scholarly journals Development of cDNA normalization system and preliminary transcription analysis of KCS genes in apple tissues

2011 ◽  
Vol 59 (3) ◽  
pp. 9-12 ◽  
Author(s):  
Zsolt Albert ◽  
Cs. Deák ◽  
A. Miskó ◽  
M. Tóth ◽  
I. Papp
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Expression Patterns ◽  
Housekeeping Genes ◽  
Apple Fruit ◽  
Rt Pcr ◽  
Specific Primers ◽  
Specific Expression ◽  
Transcription Analysis ◽  
Tissue Specific

Wax production is an important aspect of apple (Malus domestica Borkh.) fruit development from both theoretical and practical point of views. The complex molecular mechanism that controls wax biosynthesis is still widely unknown but many studies focused on this topic. We aimed to develop further the experimental framework of these efforts with a description of an improved reference genes expression system. Results in the literature show that similarities exist among the expression of some housekeeping genes of different plant species. Based on these considerations and on gene expression data from Arabidopsis thaliana, some genes in apple were assigned for analysis. EST sequences of apple were used to design specific primers for RT-PCR experiments. Isolation of intact RNA from different apple tissues and performing RT-PCR reaction were also key point in obtaining expression patterns. To monitor DNA contamination of the RNA samples, specific primers were used that amplify intron-containing sequences from the cDNA. We found that actin primers can be used for the detection of intron containing genomic DNA, and tubulin primers are good internal controls in RT-PCR experiments. We were able to make a difference between tissue-specific and tissue-independent gene-expression, furthermore we found tissue specific differences between the expression patterns of candidate genes, that are potentially involved in wax-biosynthesis. Our results show that KCS1 and KCS4 are overexpressed in the skin tissue, this could mean that these genes have skin-specific expression in apple fruit.

Novel alternative splice variant of IGF-1R and its mRNA expression patterns in BaMa and Landrace pigs

2019 ◽  
Author(s):  
Rui Yang ◽  
Lijie Dong ◽  
Songcai Liu ◽  
Yunyun Cheng ◽  
Wenzhen Wei ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Splice Variant ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Expression Level ◽  
Alternative Splice Variant ◽  
Nucleotide Polymorphism ◽  
Rt Pcr ◽  
Single Nucleotide ◽  
Muscle Tissues

The transcript variants of Insulin-like growth factor 1 receptor (IGF-1R) and their expression profiles had never been illuminated in pigs until now. Herein, we identified IGF-1R AS02 as a novel splice variant of IGF-1R gene by RT-PCR and analyzed its mRNA expression level by qRT-PCR in liver, cartilage and muscle tissues, while also detecting the single-nucleotide polymorphism (SNP) site near the splice site of the IGF-1R gene (in intron 19) of BaMa and Landrace pigs. Results demonstrated that the IGF-1R AS02 variant showed a significantly (P less than 0.05) higher expression level in cartilage than in muscle and liver across two pig breeds respectively. The expression level of the normal transcript (IGF-1R ISO01) of IGF-1R in cartilage was markedly lower than that in the other two tissues (P less than 0.05). In cartilage, IGF-1R ISO01 expression was higher in BaMa than in Landrace (P less than 0.05), while the expression level of IGF-1R AS02 was lower in BaMa than in Landrace (P less than 0.05). The SNP was detected in intron 19 of the IGF-1R gene of BaMa and Landrace pigs. These results contributed to facilitating a better understanding of IGF-1R gene in pigs.

scholarly journals Deoxynivalenol and Zearalenone: Different Mycotoxins with Different Toxic Effects in the Sertoli Cells of Equus asinus

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1898
Author(s):  
Jun-Lin Song ◽  
Guo-Liang Zhang
Keyword(s):  
Mrna Expression ◽  
Sertoli Cells ◽  
Biological Effects ◽  
Expression Patterns ◽  
Toxic Effects ◽  
Type B ◽  
Rt Pcr ◽  
Equus Asinus ◽  
The Impact

(1) Background: Deoxynivalenol (DON) and zearalenone (ZEA) are type B trichothecene mycotoxins that exert serious toxic effects on the reproduction of domestic animals. However, there is little information about the toxicity of mycotoxins on testis development in Equus asinus. This study investigated the biological effects of DON and ZEA exposure on Sertoli cells (SCs) of Equus asinus; (2) Methods: We administered 10 μM and 30 μM DON and ZEA to cells cultured in vitro; (3) Results: The results showed that 10 μM DON exposure remarkably changed pyroptosis-associated genes and that 30 μM ZEA exposure changed inflammation-associated genes in SCs. The mRNA expression of cancer-promoting genes was remarkably upregulated in the cells exposed to DON or 30 μM ZEA; in particular, DON and ZEA remarkably disturbed the expression of androgen and oestrogen secretion-related genes. Furthermore, quantitative RT-PCR, Western blot, and immunofluorescence analyses verified the different expression patterns of related genes in DON- and ZEA-exposed SCs; (4) Conclusions: Collectively, these results illustrated the impact of exposure to different toxins and concrete toxicity on the mRNA expression of SCs from Equus asinus in vitro.

Cyclooxygenase mRNA expression in human patellar tendon at rest and after exercise

2008 ◽  
Vol 294 (1) ◽  
pp. R192-R199 ◽  
Author(s):  
Todd A. Trappe ◽  
Chad C. Carroll ◽  
Bozena Jemiolo ◽  
Scott W. Trappe ◽  
Simon Døssing ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Mrna Expression ◽  
Patellar Tendon ◽  
Expression Patterns ◽  
Rt Pcr ◽  
Tendon Tissue ◽  
Intron 1 ◽  
Cox 2 ◽  
Cox 1 ◽  
Tendon Properties

Exercise has been shown to acutely elevate several metabolic processes in tendon tissue, including collagen turnover and blood flow, and chronically induce changes in tendon properties. Many of these acute metabolic responses to exercise are regulated by the cyclooxygenase (COX) enzymes. We measured the expression levels of COX-1 [variants 1 and 2 (COX-1v1 and COX-1v2)], COX-2, and the recently discovered intron 1-retaining COX-1 variants (COX-1b1, COX-1b2, and COX-1b3) at rest and after resistance exercise (RE). Patellar tendon biopsy samples were taken from six individuals (3 men and 3 women) before and 4 h after a bout of RE (3 sets of 10 repetitions at ∼70% of 1 repetition maximum) and from a separate group of six individuals (3 men and 3 women) before and 24 h after RE and analyzed by real-time RT-PCR. The COX-1 variants were the most abundant COX mRNAs before exercise and remained unchanged ( P > 0.05) after exercise. COX-2 was also expressed in tendon tissue at rest and was unchanged ( P > 0.05) after exercise. The intron 1-retaining COX-1 variants were not detectable in tendon tissue before or after exercise. COX-1 and COX-2 were expressed at much higher levels by the patellar tendon than by quadriceps skeletal muscle, although the overall COX mRNA expression patterns were similar in skeletal muscle and tendon (COX-1v2 > COX-1v1, P < 0.05; ratio of COX-1 to COX-2 ≅ 4:1). These results suggest that COX-1 and COX-2 are constitutively expressed at relatively high levels in human patellar tendon and are likely targets of COX-inhibiting drugs at rest and after physical activity.

Effect of CP-96,345 on the expression of adhesion molecules in acute pancreatitis in mice

2007 ◽  
Vol 292 (5) ◽  
pp. G1283-G1292 ◽  
Author(s):  
Hon Yen Lau ◽  
Madhav Bhatia
Keyword(s):  
Acute Pancreatitis ◽  
Mrna Expression ◽  
Adhesion Molecules ◽  
Expression Patterns ◽  
Rna Extraction ◽  
Leukocyte Recruitment ◽  
Rt Pcr ◽  
Treatment Groups ◽  
Neurokinin 1 ◽  
The Relationship

We investigated the effect of a specific neurokinin-1 receptor (NK1R) antagonist, CP-96,345, on the regulation of the expression of adhesion molecules ICAM-1, VCAM-1, E-selectin, and P-selectin as well as leukocyte recruitment during acute pancreatitis (AP). AP was induced in male Balb/C mice by 10 consecutive hourly intraperitoneal injections of caerulein. In the treatment groups, CP-96,345 was administered at 2.5 mg/kg ip either 30 min before or 1 h after the first caerulein injection. Animals were killed, and the lungs and pancreas were isolated for RNA extraction and RT-PCR or for immunohistochemical staining. mRNA expression of the four adhesion molecules was upregulated in the pancreas during AP. Treatment with CP-96,345 effectively reduced the mRNA expression of P-selectin and E-selectin but not ICAM-1 and VCAM-1. In the lung, ICAM-1, E-selectin, and P-selectin mRNA expression increased during AP. Antagonist treatment suppressed this elevation. Similar expression patterns were seen in the immunohistochemical stainings. Intravital microscopy of the pancreatic microcirculation revealed the effect of CP-96,345 on leukocyte recruitment. The present study provides important information on the relationship between NK1R activation and the regulation of adhesion molecules. Also, this study points to the differential regulation of inflammation in the pancreas and lung with AP.

scholarly journals Inflammatory Cytokine Expressions of the Subacromial Bursitis and Glenohumeral Joint Synovitis in the Patients with Full Thickness Rotator Cuff Tear

1970 ◽  
Vol 14 (2) ◽  
pp. 172-178 ◽  
Author(s):  
Sung Kyu Kim ◽  
Hyung Nam Kim ◽  
Eun Sun Moon ◽  
Keun Young Lim ◽  
Nam Young Cho ◽  
...  
Keyword(s):  
Rotator Cuff ◽  
Mrna Expression ◽  
Inflammatory Cytokines ◽  
Glenohumeral Joint ◽  
Expression Patterns ◽  
Tnf Alpha ◽  
Full Thickness ◽  
Rt Pcr ◽  
Polymerase Chain ◽  
Cox 1

PURPOSE: The purpose of this study was to compare expression patterns of inflammatory cytokines between subacromial bursitis and glenohumeral (GH) joint synovitis in full thickness rotator cuff (RC) tear patients with severe pain.MATERIALS AND METHODS: We were able to obtain enough tissue from nine subacromial bursitis and GH synovitis patients at the same time during surgery and evaluate them. We compared mRNA expression of inflammatory cytokines between the two groups using Reverse Transcription-Polymerase Chain Reaction (RT-PCR).RESULTS: Relative mean mRNA expression of IL-1beta, IL-6, TNF-alpha, COX-1, COX-2 and SDF-1 in the tissues of subacromial bursitis and GH synovitis patients did not show significant differences.CONCLUSION: GH synovitis may be another source of shoulder pain with subacromial bursitis in full thickness RC tear patients with severe pain.

scholarly journals Developmental Considerations of Sperm Protein 17 Gene Expression in Rheumatoid Arthritis Synoviocytes

2002 ◽  
Vol 9 (2) ◽  
pp. 97-102 ◽  
Author(s):  
Yuichi Takeoka ◽  
Thomas P. Kenny ◽  
Hisashi Yago ◽  
Mitsuru Naiki ◽  
M. Eric Gershwin ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Mrna Expression ◽  
Expression Patterns ◽  
Specific Protein ◽  
Differentially Expressed ◽  
Cell Binding ◽  
Specific Primers ◽  
Sperm Protein ◽  
Rheumatoid Arthritis Synoviocytes ◽  
Sperm Protein 17

Rheumatoid arthritis (RA) is an autoimmune disease characterized by proliferative synovial tissue. We used mRNA differential display and library subtraction to compare mRNA expression in RA and osteoarthritis (OA) synoviocytes. We initially compared the mRNA expression patterns in 1 female RA and 1 OA synovia and found a differentially expressed 350 bp transcript in the RA synoviocytes which was, by sequence analysis, 100% homologous to sperm protein 17 (Sp17). Moreover, the Sp17 transcript was found differentially expressed in a RA synovial library that was subtracted with an OA synovial library. Using specific primers for full length Sp17, a 1.1 kb transcript was amplified from the synoviocytes of 7 additional female RA patients, sequenced and found to 100% homologous to Sp17. Thus, we found the unexpected expression of Sp17, a thought to be gamete-specific protein, in the synoviocytes of 8/8 female RA patients in contrast to control OA synoviocytes. Interestingly, Sp17's structural relationship with cell-binding and recognition proteins, suggests that Sp17 may function in cell-cell recognition and signaling in the RA synoviocyte. Further, Sp17 could have a significant regulatory role in RA synoviocyte gene transcription and/or signal transduction. Thus, Sp17 could have an important role in RA synoviocyte proliferation or defective apoptosis. Finally, the presence of Sp17 in synoviocytes has interesting developmental considerations.

scholarly journals Seasonal and Sexual Variation in mRNA Expression of Selected Adipokine Genes Affecting Fat Deposition and Metabolism of the Emu (Dromaius Novaehollandiae)

2021 ◽  
Author(s):  
Ji Eun Kim ◽  
Darin Bennett ◽  
Kristina Wright ◽  
Kimberly M. Cheng
Keyword(s):  
Mrna Expression ◽  
Abdominal Fat ◽  
Rt Pcr ◽  
Specific Primers ◽  
Gene Markers ◽  
Reading Frame ◽  
Time Points ◽  
Genome Scanning ◽  
Dromaius Novaehollandiae ◽  
Anti Oxidant

Abstract Emus are farmed for fat production. Oil rendered from their back and abdominal fat pads has good anti-oxidant and anti-inflammatory properties and has ingredients that promote cell growth. Our objective is to examine the mRNA expression of 7 emu adipokine genes (eFABP4, eSCD1, eAdipoQ, eAdipoR1, eAdipoR2, Lept and eLepR) to identify gene markers that may help improve emu fat production. Back and abdominal fat tissues from 11 adult emus were biopsied at four time points (April, June, August and November). Total RNA was isolated and cDNA was synthesized. Gene specific primers were designed for partial cloning fragments to amplify the open reading frame of the 7 genes. Lept was not expressed in emu fat tissue. Nucleotides and amino acids sequences of the 6 expressed gene were compared with homologs from other species and phylogenetic relationships established. Seasonal mRNA expression of each gene was assessed by quantitative RT-PCR and differential expression analysed by the 2−ΔΔCT method. The temporal mRNA expression pattern of the genes and the fat gain (kg) between time points association with gene expression level were determined. More whole-genome scanning studies are needed to develop novel molecular markers that can be applied to improve fat production in emus.

Genexpressionsmuster von fibrinolytischen Faktoren des Urokinase-Plasmin-Systems bei Dermatoliposklerose

Phlebologie ◽  
1999 ◽  
Vol 28 (01) ◽  
pp. 1-6 ◽  
Author(s):  
Ch. Stetter ◽  
E. Schöpf ◽  
J. Norgauer ◽  
W. Vanscheidt ◽  
Y. Herouy
Keyword(s):  
Mrna Expression ◽  
De Novo ◽  
Ulcus Cruris ◽  
Ulcus Cruris Venosum ◽  
Rt Pcr ◽  
Fand Sich ◽  
Pai 1

ZusammenfassungDie Dermatoliposklerose (DLS) entwickelt sich als Folge einer progredienten primären Varikosis oder eines postthrombotischen Syndroms (PTS). Trotz bestehender Hinweise auf eine veränderte intravasale fibrinolytische Aktivität bei der chronisch-venösen Insuffizienz (CVI), wurden bisher fibrinolytische Faktoren im perivaskulären Gewebe nicht untersucht. Kürzlich zeigten wir, daß bei Dermatoliposklerose Matrix-Metalloproteinasen exprimiert und aktiviert werden. Da spezifische fibrinolytische Faktoren wichtige Haupteffektoren der Matrix-Metalloproteinasenaktivierung sind, untersuchten wir kürzlich die Genexpression der Plasminogenaktivatoren vom Urokinasetyp (uPA) und vom Gewebetyp (tPA), des Urokinase-Rezeptor (uPA-R) sowie der Plasminogenaktivator-Inhibitoren (PAI-1 und PAI-2) in Gewebsbiopsien von Patienten mit Dermatoliposklerose. Zum Nachweis verwandten wir dabei die Technik der reversen Transkription und Polymerase-Kettenreaktion (RT-PCR). Es fand sich in allen Hautproben (n = 21) eine signifikant erhöhte mRNA-Expression von uPA und uPA-R im Vergleich zu gesunder Haut (n = 12). Dagegen konnte kein signifikanter Unterschied für mRNA-Transkripte von tPA, PAI-1 und PAI-2 nachgewiesen werden. Die Dermatoliposklerose zeichnet sich somit durch erhöhte transkriptionelle Expression von uPA und uPA-R aus. Eine gesteigerte De-novo-Synthese von uPA und uPA-R könnte daher bei der Aktivierung von Matrix-Metalloproteinasen und entsprechend in der Pathogenese des Ulcus cruris venosum eine zentrale Rolle spielen.

scholarly journals Ex vivo mRNA expression of toll-like receptors during latent tuberculosis infection

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Birhan Alemnew ◽  
Soren T. Hoff ◽  
Tamrat Abebe ◽  
Markos Abebe ◽  
Abraham Aseffa ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Expression Patterns ◽  
Fold Change ◽  
Mrna Levels ◽  
Healthy Children ◽  
Toll Like Receptors ◽  
Relative Expression ◽  
Latently Infected ◽  
Latent Tb ◽  
Latent Tb Infection

Abstract Background Understanding immune mechanisms, particularly the role of innate immune markers during latent TB infection remains elusive. The main objective of this study was to evaluate mRNA gene expression patterns of toll-like receptors (TLRs) as correlates of immunity during latent TB infection and further infer their roles as potential diagnostic biomarkers. Methods Messenger RNA (mRNA) levels were analysed in a total of 64 samples collected from apparently healthy children and adolescents latently infected with tuberculosis (n = 32) or non-infected (n = 32). Relative expression in peripheral blood of selected genes encoding TLRs (TLR-1, TLR-2, TLR-4, TLR-6 and TLR-9) was determined with a quantitative real-time polymerase chain reaction (qRT-PCR) using specific primers and florescent labelled probes and a comparative threshold cycle method to define fold change. Data were analysed using Graph-Pad Prism 7.01 for Windows and a p-value less than 0.05 was considered statistically significant. Results An increased mean fold change in the relative expression of TLR-2 and TLR-6 mRNA was observed in LTBI groups relative to non-LTBI groups (p < 0.05), whereas a slight fold decrease was observed for TLR-1 gene. Conclusions An increased mRNA expression of TLR-2 and TLR-6 was observed in latently infected individuals relative to those non-infected, possibly indicating the roles these biomarkers play in sustenance of the steady state interaction between the dormant TB bacilli and host immunity.

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