The Two mRNAS Encoding Rat Intestinal Alkaline Phosphatase Represent Two Distinct Nucleotide Sequences

1992 ◽  
Vol 38 (12) ◽  
pp. 2506-2509 ◽  
Author(s):  
M J Engle ◽  
D H Alpers
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Duodenal Mucosa ◽  
Differential Regulation ◽  
Untranslated Regions ◽  
Terminal Sequence ◽  
Intestinal Alkaline Phosphatase ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Fat Feeding

Abstract Rat duodenal mucosa contains two mRNAs (2.7 and 3.0 kb) encoding intestinal alkaline phosphatase (IAP), but the mechanism for their production has not been clear. By means of the polymerase chain reaction (PCR), we isolated a fragment that identifies a second rat IAP (rIAP-II), differing from the rIAP-I sequence in the coding and 3' untranslated regions and encoding a COOH-terminal sequence predicted to be hydrophilic. By means of probes unique to each sequence, rIAP-I identified the 2.7-kb mRNA, and rIAP-II the 3.0-kb mRNA. The different structures and differential regulation of the mRNAs after fat feeding demonstrate the presence of two rat IAP transcripts.

scholarly journals Transient benign hyperphosphatasemia due to COVID-19: the first case report

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Tugba Erat ◽  
Müge Atar ◽  
Tugba Kontbay
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Case Report ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Benign Condition ◽  
Case Presentation ◽  
Normal Ranges ◽  
First Case ◽  
Polymerase Chain

AbstractObjectivesCoronavirus disease (COVID-19) rapidly spread worldwide in a few months and was declared as a worldwide pandemic by WHO in March 2020. Transient benign hyperphosphatasemia (THI) is a benign condition associated with marked elevation of alkaline phosphatase (ALP) without any other kidney, bone, and liver pathologies.Case presentationHerein, we report a previously healthy 16-month-old female patient who developed a secondary transient benign hyperphosphatasemia associated with SARS-CoV-2. Patient whole family’s SARS-CoV-2 real-time reverse transcription-polymerase chain reaction (RT-PCR) results were positive. Since THI is a diagnosis of exclusion, other reasons that may cause ALP elevation should be ruled out. ALP activity decreased and turned to normal ranges within the following month. THI has been reported to be in association with various conditions. Its relationship with many viruses has been reported previously.ConclusionsIf ALP elevation is detected in patients with COVID 19 due to the increasing number of infections, THI should be considered if there is no other accompanying pathology.

scholarly journals Vitamin D Enhanced the Osteogenic Differentiation of Cell Spheroids Composed of Bone Marrow Stem Cells

Medicina ◽  
2021 ◽  
Vol 57 (11) ◽  
pp. 1271
Author(s):  
Hyun-Jin Lee ◽  
Young-Min Song ◽  
Seunghoon Baek ◽  
Yoon-Hee Park ◽  
Jun-Beom Park
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Vitamin D ◽  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Cellular Viability ◽  
Chain Reaction ◽  
Cell Spheroids ◽  
Polymerase Chain

Background and Objectives: Vitamin D is a bone modulator widely used in regenerative medicine. This study aimed to analyze the effects of vitamin D on the osteogenic differentiation and mineralization of human mesenchymal stem cells. Materials and Methods: Spheroids were fabricated using human bone marrow-derived stem cells, and were cultured in the presence of vitamin D at concentrations of 0, 0.1, 1, 10, and 100 nM. Stem cell spheroids were fabricated and the morphological evaluation was conducted on days 1, 3, 7 and 14. Determination of qualitative cellular viability was performed with Live/Dead Kit assay on days 1 and 7. Quantitative cellular viability was evaluated with Cell Counting Kit-8 on days 1, 3, 7, and 14. To analyze the osteogenic differentiation of cell spheroids, alkaline phosphatase activity assays were performed with commercially available kit on days 7 and 14. Real-time polymerase chain reaction was used to determine the expression levels of RUNX2, BSP, OCN, and COL1A1 on days 7 and 14. Results: The stem cells produced well-formed spheroids, and addition of vitamin D did not result in any noticeable changes in the shape. The addition of vitamin D did not significantly change the diameter of the spheroids at 0, 0.1, 1, 10, or 100 nM concentrations. Quantitative cell viability results from days 1, 3, 7 and 14 showed no significant difference between groups (p > 0.05). There was significantly higher alkaline phosphatase activity in the 0.1 nM group when compared with the control group on day 14 (p < 0.05). Real-time polymerase chain reaction results demonstrated that the mRNA expression levels of RUNX2, OCN, and COL1A1 were significantly increased when vitamin D was added to the culture. Conclusions: Based on these findings, we concluded that vitamin D could be applied to the increased osteogenicity of stem cell spheroids.

Osteogenic Activity of MG63 Cells on Hydroxyapatite/Collagen Nanocompsosite under Pressure/perfusion Culture

2007 ◽  
Vol 330-332 ◽  
pp. 317-320 ◽  
Author(s):  
Teruaki Yoshida ◽  
Masanori Kikuchi ◽  
Yoshihisa Koyama ◽  
Kazuo Takakuda
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Perfusion Culture ◽  
Collagen Sponge ◽  
Perfusion Technique ◽  
Chain Reaction ◽  
Osteogenic Activity ◽  
Self Organized ◽  
Mg63 Cells ◽  
Polymerase Chain

The purpose of this study is to analyze osteogenic activity of human osteoblastic MG63 cells on the bone-like self-organized hydroxyapatite/collagen (HAp/Col) nanocomposite sponge cultured by a pressure/perfusion technique using collagen sponge as a control. Histological analyses, alkaline phosphatase (AlkP) protein analysis and real-time polymerase chain reaction (PCR) analyses for AlkP and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to evaluate the osteogenic activity of MG63. The MG63 cells were attached well and showed good proliferation on the HAp/Col sponge as well as in the control. The MG63 cells on HAp/Col sponge demonstrated higher osteogenic activity than those on the control. The results suggested that the HAp/Col sponge is expected to be a good scaffold for bone tissue engineering.

scholarly journals Two alkaline phosphatase genes are expressed during early development in the mouse embryo

Development ◽  
1990 ◽  
Vol 110 (2) ◽  
pp. 555-564 ◽  
Author(s):  
A.C. Hahnel ◽  
D.A. Rappolee ◽  
J.L. Millan ◽  
T. Manes ◽  
C.A. Ziomek ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Germ Cells ◽  
Inner Cell Mass ◽  
Primordial Germ Cells ◽  
Cell Mass ◽  
Mouse Embryos ◽  
Preimplantation Development ◽  
Chain Reaction ◽  
Polymerase Chain

Alkaline phosphatase (AP) activity is stage specific in mouse embryos and may be associated with compaction and separation of trophectoderm from inner cell mass in preimplantation development. We previously sequenced a cDNA and two mouse AP genes that could contribute to the AP activity in embryos. Oligonucleotide primers were constructed from the three sequences and used in the reverse transcription-polymerase chain reaction technique to establish that two of the three AP isozymes are transcribed during preimplantation development. The predominant transcript (E-AP) is from a gene highly homologous to the human tissue-specific APs, but different from the mouse intestinal AP. Tissue non-specific (TN) AP also is transcribed, but there is approximately 10 times less TN-AP than E-AP transcript. The TN-AP isozyme is the predominant transcript of 7 to 14 day embryos and primordial germ cells. A switch in predominance from E-AP to TN-AP must occur during early postimplantation development. This study establishes a framework for experiments to determine the functions of the two isozymes during preimplantation development.

Improved specificity ofTrypanosoma cruziidentification by polymerase chain reaction using an oligonucleotide derived from the amino-terminal sequence of a Tc24 protein

Parasitology ◽  
1995 ◽  
Vol 111 (5) ◽  
pp. 581-590 ◽  
Author(s):  
A. Taibi ◽  
A. Guevara-Espinoza ◽  
R. Schöneck ◽  
B. Yahiaoui ◽  
A. Ouaissi
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Value ◽  
Terminal Sequence ◽  
Chain Reaction ◽  
Blood Samples ◽  
Specific Diagnosis ◽  
Amino Terminal ◽  
Polymerase Chain ◽  
Amino Terminal Domain ◽  
Amino Terminal Sequence

SUMMARYIn the present study, the diagnostic value ofTrypanosoma cruzirecombinant protein (Tc24) was examined. Although antibodies against Tc24 were detected during natural and experimentalT. cruziinfections, specificity studies revealed that sera fromT. rangeli-infected mice also recognized to some extent Tc24 protein. In addition, sera from Tc24-immunized mice reacted against a 21 kDa polypeptide inT. rangeliextracts. Detailed analysis of the antibody response against 20—40 peptide localized in the Tc24 amino-terminal domain suggests that this sequence is not expressed byT. rangeli21 kDa antigen. Therefore, the PCR reaction using oligonucleotides corresponding to a 20–26 peptide clearly demonstrated the specificity of the oligoprobes forT. cruziidentification. Positive signals were also found when using blood samples fromT. cruzi-infected mice. Taken together, these results suggest that the PCR-based 20–26 assay may be useful in the specific diagnosis of Chagas' disease.

scholarly journals Enhanced osteogenic differentiation of alendronate-conjugated nanodiamonds for potential osteoporosis treatment

2021 ◽  
Vol 25 (1) ◽  
Author(s):  
Guk Young Ahn ◽  
Sung-Eun Kim ◽  
Tae Hoon Yun ◽  
Inseong Choi ◽  
Daewon Park ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
Osteoporosis Treatment ◽  
Bone Diseases ◽  
Calcium Deposition ◽  
Therapeutic Effects ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background Alendronate (Alen) is promising material used for bone-targeted drug delivery due to its high bone affinity and therapeutic effects on bone diseases. In addition, Alen can enhance the osteogenic differentiation of osteoblastic cell. Recently, nanodiamonds (NDs) with hardness, non-toxicity, and excellent biocompatibility are employed as promising materials for carrier systems and osteogenic differentiation. Therefore, we prepared Alen-conjugated NDs (Alen-NDs) and evaluated their osteogenic differentiation performances. Methods Alen-NDs were synthesized using DMTMM as a coupling reagent. Morphological change of Mouse calvaria-derived preosteoblast (MC3T3-E1) treated with Alen-NDs was observed using the confocal microscope. The osteogenic differentiation was confirmed by cell proliferation, alkaline phosphatase (ALP), calcium deposition, and real-time polymerase chain reaction assay. Results Alen-NDs were prepared to evaluate their effect on the proliferation and differentiation of osteoblastic MC3T3-E1 cells. The Alen-NDs had a size of about 100 nm, and no cytotoxicity at less than 100 μg/mL of concentration. The treatment of NDs and Alen-NDs reduced the proliferation rate of MC3T3-E1 cells without cell death. Confocal microscopy images confirmed that the treatment of NDs and Alen-NDs changed the cellular morphology from a fibroblastic shape to a cuboidal shape. Flow cytometry, alkaline phosphatase (ALP) activity, calcium deposition, and real-time polymerase chain reaction (RT-PCR) confirmed the higher differentiation of MC3T3-E1 cells treated by Alen-NDs, compared to the groups treated by osteogenic medium and NDs. The higher concentration of Alen-ND treated in MC3T3-E1 resulted in a higher differentiation level. Conclusions Alen-NDs can be used as potential therapeutic agents for osteoporosis treatment by inducing osteogenic differentiation.

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