Sodium Butyrate Enhancement of Protein Expression in Hela Cells: Follow-Up of a Chimeric Reporter Gene Behaviour for Process Development

2005 ◽  
pp. 625-627
Author(s):  
C. Bassens ◽  
P. Rigaux ◽  
V. Hendrick ◽  
M. Cherlet ◽  
K. Sato ◽  
...  
Keyword(s):  
Protein Expression ◽  
Reporter Gene ◽  
Sodium Butyrate ◽  
Hela Cells ◽  
Process Development

Expression of recombinant protein in cho and hela cells and its follow-up using EGF reporter gene

2006 ◽  
pp. 55-59
Author(s):  
V. Hendrick ◽  
D. Ribeiro Sousa ◽  
A. R. dos Santos Pedregal ◽  
C. Bassens ◽  
P. Rigaux ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Reporter Gene ◽  
Hela Cells

scholarly journals Inhibition of p38 Mitogen-Activated Protein Kinase Impairs Mayaro Virus Replication in Human Dermal Fibroblasts and HeLa Cells

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1156
Author(s):  
Madelaine Sugasti-Salazar ◽  
Yessica Y. Llamas-González ◽  
Dalkiria Campos ◽  
José González-Santamaría
Keyword(s):  
Protein Expression ◽  
Hela Cells ◽  
Plaque Assay ◽  
Dermal Fibroblasts ◽  
Dependent Manner ◽  
Human Dermal Fibroblasts ◽  
Mitogen Activated Protein ◽  
Mayaro Virus ◽  
The Impact ◽  
Dose Dependent

Mayaro virus (MAYV) hijacks the host’s cell machinery to effectively replicate. The mitogen-activated protein kinases (MAPKs) p38, JNK, and ERK1/2 have emerged as crucial cellular factors implicated in different stages of the viral cycle. However, whether MAYV uses these MAPKs to competently replicate has not yet been determined. The aim of this study was to evaluate the impact of MAPK inhibition on MAYV replication using primary human dermal fibroblasts (HDFs) and HeLa cells. Viral yields in supernatants from MAYV-infected cells treated or untreated with inhibitors SB203580, SP600125, U0126, or Losmapimod were quantified using plaque assay. Additionally, viral protein expression was analyzed using immunoblot and immunofluorescence. Knockdown of p38⍺/p38β isoforms was performed in HDFs using the PROTACs molecule NR-7h. Our data demonstrated that HDFs are highly susceptible to MAYV infection. SB203580, a p38 inhibitor, reduced MAYV replication in a dose-dependent manner in both HDFs and HeLa cells. Additionally, SB203580 significantly decreased viral E1 protein expression. Similarly, knockdown or inhibition of p38⍺/p38β isoforms with NR-7h or Losmapimod, respectively, affected MAYV replication in a dose-dependent manner. Collectively, these findings suggest that p38 could play an important role in MAYV replication and could serve as a therapeutic target to control MAYV infection.

scholarly journals C-MYC Protein Expression and High Ki-67 Proliferative Index are Predictives of Disease Relapse in Diffuse Large B Cell Lymphoma

2021 ◽  
Vol 6 (1) ◽  
pp. 15-20
Author(s):  
Mahmoud Tag El-Hussien ◽  
Nadia Mokhtar ◽  
Eman Naguib Khorshed
Keyword(s):  
Protein Expression ◽  
B Cell ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Statistical Prediction ◽  
Proliferative Index ◽  
Ki 67 ◽  
Large B Cell Lymphoma ◽  
Large B Cell

Objective: To evaluate the status of C-MYC protein expression and Ki-67 proliferative index and to clarify their role in predicting relapse of diffuse large B cell lymphoma (DLBL). Materials and Methods: A retrospective study conducted on 50 cases diagnosed as DLBL in a 3 years’ time period from January 2014 till December 2016, collected from the archive of Pathology Departments of the National Cancer Institute Cairo - Egypt, Misr University for Science and Technology and private labs of authors. The diagnosis of DLBL for all cases, both nodal and extranodal, was confirmed by histopathologic examination and immunophenotyping. Automated immunohistochemical staining using antibodies against C-MYC protein and MIB-1 was used to evaluate the C-MYC expression in tumor cells and to assess their proliferative ability by calculating Ki-67 labelling index. The relation between the percentage of C-MYC protein expression, Ki-67 proliferative index, clinical data and the relapse status during the follow up period were analyzed. Results: A total of 50 cases of DLBL in both nodal and extra-nodal sites were included. Twenty-three cases (46%) were expressing the C-MYC protein, and 29 cases (58%) showed high Ki-67 proliferative index. Twenty-two cases (44%) relapsed during the follow-up period. Positive C-MYC protein expression was significantly associated with high Ki-67 proliferative index. C-MYC protein expression and high Ki-67 proliferative index were independently associated with disease relapses in 81.8% and 86.4% of cases respectively. Cases with combined C-MYC protein expression and high Ki-67 proliferative index showed statistical prediction of relapse in 81.8% of cases. Conclusion: C-MYC protein expression and high Ki-67 proliferative index were independently associated with relapse of diffuse large B cell lymphoma. Furthermore, the combined positive C-MYC protein expression and high Ki-67 proliferative index is better than a single positive test in predicting relapses among DLBL patients.

scholarly journals Deciding the operation type according to mismatch repair status among hereditary nonpolyposis colorectal cancer patients: should a tailored approach be applied, or does one size fit all?

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Chun-Kai Liao ◽  
Yueh-Chen Lin ◽  
Yu-Jen Hsu ◽  
Yih-Jong Chern ◽  
Jeng-Fu You ◽  
...  
Keyword(s):  
Overall Survival ◽  
Protein Expression ◽  
Mismatch Repair ◽  
Multivariate Analyses ◽  
Extent Of Resection ◽  
Significant Difference ◽  
Tailored Approach ◽  
Mismatch Repair Status ◽  
Colorectal Cancer Patients

Abstract Background Although extended colectomy (EC) was recommended for HNPCC patients, previous studies did not show significantly improved overall survival. Immunohistochemical (IHC) stain of mismatch repair (MMR) gene protein expression is now a feasible and reliable test clinically. Therefore, we tried to investigate whether we could use MMR IHC stain to select operation types in HNPCC patients. Patients and methods Between 1995 and 2013, 186 HNPCC patients were collected. Status of MMR protein expression, perioperative clinic-pathological variables and post-operative follow up status were analyzed by multivariate analyses. Results Sixty-five percent (121 of 186) patients of these HNPCC patients demonstrated loss of at least one MMR protein. There were several significant differences existing between deficient MMR (dMMR) and proficient MMR (pMMR) subgroups in terms of clinic-pathological characteristics. With the average follow-up duration of 93.9 months, we observed significantly high risk of developing metachronous CRC between SC and EC subgroups (crude rate 8.5% vs. 0%, p = 0.035). However, no significant difference was observed among the presence of extra-colonic tumors (12.4% vs. 5.8%, p = 0.284). The positive and negative prediction rate of metachronous CRC in dMMR subgroup was 12.8 and 87.2% while 1.9 and 98.1% in the pMMR subgroup. Survival outcomes were significantly affected by MMR status and resection types by multivariate analysis. Significantly better OS in dMMR subgroup (HR = 0.479, 95% CI: 0.257–0.894, p = 0.021) comparing with pMMR subgroup was observed. However, significant improved DFS (HR = 0.367, 95% CI: 0.172–.0787, p = 0.010) but not significant for OS (HR = 0.510, 95% CI: 0.219–1.150, p = 0.103) for EC subgroup compared with SC subgroup. Differences existing among different subgroups by combing extent of resection and MMR status. In dMMR subgroup, SC, compared with EC, demonstrated significantly worse DFS by multivariate analyses (HR = 3.526, 95% CI: 1.346–9.236, p = 0.010) but not for OS (HR = 2.387, 95% CI: 0.788–7.229, p = 0.124), however, no significantly differences of OS and DFS in pMMR subgroup between SC and EC were found. Conclusions Significantly better overall survival and higher rate of metachronous CRC exist in dMMR subgroup of HNPCC patients comparing with pMMR subgroup. Extended colectomy significantly improved DFS and was thus recommended for dMMR subgroup but not pMMR subgroup of HNPCC patients.

Abstract P38: The Great Lakes Regional Stroke Network Swallow Screen Project

2011 ◽  
Vol 4 (suppl_1) ◽  
Author(s):  
Angela Hedworth ◽  
Silvina B Tonarelli ◽  
Gabriela Vazquez
Keyword(s):  
Great Lakes ◽  
Aspiration Pneumonia ◽  
Process Development ◽  
Work Group ◽  
National Level ◽  
Evidence Base ◽  
Screening Tools ◽  
Screen Tool ◽  
Program Coordinators

Background and purpose A formal dysphagia protocol including a swallow screen can decrease risk of aspiration pneumonia in stroke patients. To date the varying assortment of tools and protocols available is overwhelming. The aim of this project was to assess and summarize the current swallow screen tools and processes utilized by hospitals in the Great Lakes Region and to determine if participating in a work group impacted their swallow screen process. Methods A work group was created to evaluate screening tools and processes. The group consisted of nurses, speech language pathologists, stroke program coordinators and neuroscience administrators from over 35 hospitals in the Midwest. One year after the group was formed, a follow up evaluation was sent to participants to determine any changes. Results Initial findings revealed a high degree of variance between facilities' tools and processes. Water was used in 86%, 60% included a substance thicker than water, and 6% specifically state the use of straws. The follow-up evaluation had a 74% (n=26) response rate. 48% of those respondents reported modifications to their swallow screen tool as a result of participating in the work group; 33% adopted of a new tool from the literature; 48% reported changing their swallow screen process; and 56% changed their swallow screen training. 44% do not monitor the correlation between swallow screening and aspiration pneumonia. Conclusions Respondents reported the most beneficial aspects of this project was the opportunity to see other swallow screens, providing an evidence base for screens in use, and prompting an internal evaluation of their screen/process. Despite The Joint Commission no longer requiring dysphagia in its data collection, all reported they will continue to collect that data. All facilities shared the same challenges with compliance, on-going education, and documentation. Opportunities for improvement were identified: additional analysis and synthesis of screening tools and methodology at the national level; better understanding and process development between speech language pathology and nursing to standardize a universal swallow screen tool.

scholarly journals Discovery of increased epidermal DNAH10 expression after regeneration of dermis in a randomized with-in person trial — reflections on psoriatic inflammation

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Heli Lagus ◽  
Mariliis Klaas ◽  
Susanna Juteau ◽  
Outi Elomaa ◽  
Juha Kere ◽  
...  
Keyword(s):  
Protein Expression ◽  
Psoriatic Skin ◽  
Dermal Matrix ◽  
Thickness Skin ◽  
Dermal Regeneration ◽  
Unaffected Skin ◽  
One Year ◽  
To Receive

AbstractBecause molecular memories of past inflammatory events can persist in epidermal cells, we evaluated the long-term epidermal protein expression landscapes after dermal regeneration and in psoriatic inflammation. We first characterized the effects of two dermal regeneration strategies on transplants of indicator split-thickness skin grafts (STSGs) in ten adult patients with deep burns covering more than 20% of their body surface area. After fascial excision, three adjacent areas within the wound were randomized to receive a permanent dermal matrix, a temporary granulation-tissue-inducing dressing or no dermal component as control. Control areas were covered with STSG immediately, and treated areas after two-weeks of dermis formation. Epidermis-dermis-targeted proteomics of one-year-follow-up samples were performed for protein expression profiling. Epidermal expression of axonemal dynein heavy chain 10 (DNAH10) was increased 20-fold in samples having had regenerating dermis vs control. Given the dermal inflammatory component found in our dermal regeneration samples as well as in early psoriatic lesions, we hypothesized that DNAH10 protein expression also would be affected in psoriatic skin samples. We discovered increased DNAH10 expression in inflammatory lesions when compared to unaffected skin. Our results associate DNAH10 expression with cell proliferation and inflammation as well as with the epidermal memory resulting from the previous regenerative signals of dermis. This study (ISRCTN14499986) was funded by the Finnish Ministry of Defense and by government subsidies for medical research.

scholarly journals Aplastic anemia and paroxysmal nocturnal hemoglobinuria: search for a pathogenetic link

Blood ◽  
1995 ◽  
Vol 85 (5) ◽  
pp. 1354-1363 ◽  
Author(s):  
A Griscelli-Bennaceur ◽  
E Gluckman ◽  
ML Scrobohaci ◽  
P Jonveaux ◽  
T Vu ◽  
...  
Keyword(s):  
Protein Expression ◽  
Aplastic Anemia ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Antithrombin Iii ◽  
Clinical Symptoms ◽  
Flow Cytometric Analysis ◽  
Normal Subjects ◽  
Initial Study ◽  
Pnh Clone

The association of paroxysmal nocturnal hemoglobinuria (PNH) and aplastic anemia (AA) raises the yet unresolved questions as to whether these two disorders are different forms of the same disease. We compared two groups of patients with respect to cytogenetic features, glycosylphosphatidylinositol (GPI)-linked protein expression, protein C/protein S/thrombomodulin/antithrombin III activity, and PIG-A gene expression. The first group consisted of eight patients with PNH (defined as positive Ham and sucrose tests at diagnosis), and the second, 37 patients with AA. Twelve patients with AA later developed a PNH clone. Monoclonal antibodies used to study GPI-linked protein expression (CD14 [on monocytes], CD16 [on neutrophils], CD48 [on lymphocytes and monocytes], CD67 [on neutrophils and eosinophils], and, more recently, CD55, CD58, and CD59 [on erythrocytes]) were also tested on a cohort of 20 normal subjects and five patients with constitutional AA. Ham and sucrose tests were performed on the same day as flow- cytometric analysis. Six of 12 patients with AA, who secondarily developed a PNH clone, had clinical symptoms, while all eight patients with PNH had pancytopenia and/or thrombosis and/or hemolytic anemia. Cytogenetic features were normal in all but two patients. Proteins C and S, thrombomodulin, and antithrombin III levels were within the normal range in patients with PNH and in those with AA (with or without a PNH clone). In patients with PNH, CD16 and CD67 expression were deficient in 78% to 98% of the cells and CD14 in 76% to 100%. By comparison, a GPI-linked defect was detected in 13 patients with AA, affecting a mean of 32% and 33% of CD16/CD67 and CD14 cell populations, respectively. Two of three tested patients with PNH and 1 of 12 patients with AA had a defect in the CD48 lymphocyte population. In a follow-up study of our patient cohort, we used the GPI-linked molecules on granulocytes and monocytes investigated earlier and added the study of CD55, CD58, and CD59 on erythrocytes. Two patients with PNH and 14 with AA were studied for 6 to 13 months after the initial study. Among patients with AA, four in whom no GPI-anchoring defect was detected in the first study had no defect in follow-up studies of all blood-cell subsets (including erythrocytes). Analysis of granulocytes, monocytes, and erythrocytes was performed in 7 of 13 AA patients in whom affected monocytes and granulocytes were previously detected. A GPI-anchoring defect was detected on erythrocytes in five of six.(ABSTRACT TRUNCATED AT 400 WORDS)

Induction of alkaline phosphatase activity in HeLa cells by sodium butyrate

In Vitro ◽  
1979 ◽  
Vol 15 (11) ◽  
pp. 861-864 ◽  
Author(s):  
Walker Wharton ◽  
Cathryn A. Hart ◽  
Barry Goz
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Sodium Butyrate ◽  
Hela Cells

Platelet Microrna-mRNA Co-Expression Profiles Correlate with Platelet Reactivity

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2016-2016
Author(s):  
Srikanth Nagalla ◽  
Chad Shaw ◽  
Xianguo Kong ◽  
Lin Ma ◽  
Altaf A. Kondkar ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Protein Expression ◽  
Reporter Gene ◽  
Microarray Analysis ◽  
Individual Variation ◽  
Healthy Subjects ◽  
Platelet Reactivity ◽  
Human Platelets ◽  
Differentially Expressed ◽  
Cell Physiology

Abstract Abstract 2016 Background: There is extreme inter-individual variation in platelet reactivity, which likely impacts the variation in both risk and clinical outcome of ischemic vascular disease since platelet hyperreactivity has prospectively been shown to be a risk for recurrent coronary syndromes. Although heritability strongly influences the inter-individual variation in platelet reactivity, there is a lack of understanding of the molecular and genetic mechanisms responsible for this variability. To understand some of these mechanisms, we have previously performed mRNA microarray analysis on platelets of subjects with differing levels of platelet reactivity. We showed a differentially expressed (DE) transcript (VAMP8) was associated with platelet reactivity. Intriguingly, we identified a possible role for microRNA (miRNA)-96 in the regulation of VAMP8 mRNA and protein expression. MiRNAs regulate numerous aspects of normal cell physiology and cause disease by altering protein expression, and recent data demonstrate a role for miRNAs in both normal and diseased human megakaryocytopoiesis. Although others and we have observed miRNAs in platelets, their biology is largely unexplored. Aims: To test whether platelet miRNA levels were associated with platelet reactivity in 19 healthy subjects. Because we had previously obtained platelet mRNA profile data on these 19 subjects, we also had a unique opportunity to test for relationships between differentially expressed miRNAs and target DE mRNAs. Methods: MiRNA microarray analysis was performed on leukocyte depleted platelets from 19 healthy subjects with marked variability in platelet responsiveness. Bioinformatics approaches were used analyze the miRNA data in platelets. Subsequently transfection experiments in cell lines to assess miRNA knockdown of target gene products and reporter gene assays were used for functional assessment of miRNA binding to 3’UTRs of the target genes. Results: We found that human platelets express 284 miRNAs, some at very high levels. Unsupervised hierarchical clustering of miRNA profiles resulted in two groups of subjects that appeared to cluster by platelet aggregation phenotypes. Seventy-four miRNAs were differentially expressed between subjects grouped according to platelet aggregation to epinephrine, a subset of which predicted the platelet reactivity response. We profiled miRNAs from HEL and Meg-01 cells and found a strong correlation between normal human platelets and both HEL cells and Meg-01 cells. Using whole-genome mRNA expression data on these same 19 subjects, we computationally generated a high-priority list of miRNA-mRNA pairs where the differentially expressed platelet miRNAs had binding sites in 3’UTRs of differentially expressed mRNAs, and the levels were negatively correlated. From this list, three miRNA-mRNA pairs (miR-200b:PRKAR2B, miR-495:KLHL5 and miR-107:CLOCK) were selected, and all three miRNAs knocked down the protein expression of the target mRNA. Co-transfection experiments using reporter gene constructs engineered to contain the candidate mRNA 3’UTR and corresponding miRNA demonstrated that the miRNA of interest directly targeted the 3’UTR of the candidate mRNA. Conclusions: Results from this study demonstrated (1) platelet miRNAs are able to repress expression of platelet proteins; (2) platelet miRNA profiles are associated with and may predict platelet reactivity and (3) bioinformatic approaches can successfully identify functional miRNAs in platelets. Our findings suggest that selected platelet miRNAs may have potential as biomarkers for vascular thrombosis. It will be important to consider the repertoire and levels of miRNAs when attempting to elucidate the molecular mechanisms responsible for inter-individual variation in platelet reactivity. Disclosures: No relevant conflicts of interest to declare.

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