Characterisation of a human bilirubin UDP-glucuronosyltransferase stably expressed in hamster lung fibroblast cell cultures

AbstractA cDNA encoding a human bilirubin UDP-glucuronosyltransferase has been isolated and stably expressed in Chinese hamster V79 lung fibroblast cell line. Western blotting of cell homogenates with anti-UGT antibody revealed a highly expressed protein of approx. 55.5 kDa in size. The expressed enzyme specifically catalysed the formation of bilirubin mono- and diglucuronides, and also catalysed the glucuronidation of two phenolic compounds, which are good substrates for other human UGT isoenzymes, at low rates

Characterization of Mononucleated Human Peripheral Blood Cells

Unspecialized cells that can renew themselves and give rise to multiple differentiated cell types are termed stem cells. The objective of this study was to characterize and investigate, through molecular and biochemical analyses, the stemness of cells derived from isolated mononucleated cells that originated from peripheral blood. The isolated mononucleated cells were separated according to their physical characteristics (adherent and suspension), after 4 to 7 days into a 14-day culturing period in complete medium. Our results revealed that adherent and suspension cells were positive for mesenchymal stem cell (MSC) and hematopoietic stem cell (HSC) markers, respectively. Differentiation of adherent cells into osteoblasts was associated with expression of the OPN gene and increasing ALP enzyme activity, while differentiation of suspension cells into osteoclasts was associated with expression of the TRAP gene and increasing TRAP enzyme activity. In conclusion, molecular and biochemical analyses showed that mononucleated cells consist of MSC (adherent) and HSC (suspension), and both cell types are able to differentiate into specialized cells from their respective lineage: osteoblast (MSC) and osteoclast (HSC)

Stem Cell Heterogeneity of Mononucleated Cells from Murine Peripheral Blood: Molecular Analysis

The main purpose of this paper was to determine the heterogeneity of primary isolated mononucleated cells that originated from the peripheral blood system by observing molecular markers. The isolated cells were cultured in complete medium for 4 to 7 days prior to the separation of different cell types, that is, adherent and suspension. Following a total culture time of 14 days, adherent cells activated the Cd105 gene while suspension cells activated the Sca-1 gene. Both progenitor markers, Cbfa-1 and Ostf-1, were inactivated in both suspension and adherent cells after 14-day culture compared to cells cultured 3 days in designated differentiation medium. In conclusion, molecular analyses showed that primary mononucleated cells are heterogeneous, consisting of hematopoietic stem cells (suspension) and mesenchymal stem cells (adherent) while both cells contained no progenitor cells

Molecular Markers of Dental Pulp Tissue during Orthodontic Tooth Movement: A Pilot Study

Three specific orthodontic tooth movement genes, that is, FCRL1, HSPG2, and LAMB2 were detected at upper first premolar (with appliance) dental pulp tissue by using GeneFishing technique as compared to lower first premolar (without appliance). These three differentially expressed genes have the potential as molecular markers during orthodontic tooth movement by looking at molecular changes of pulp tissue

Penyerapan akar gigi apeks luaran hasil rawatan ortodontik pada enam dan 12 bulan

Penyerapan akar gigi apeks luaran (PAAL) adalah salah satu kesan negatif semasa rawatan ortodontik selain daripada gigi yang mengalami trauma. Objektif kajian ini adalah untuk melihat hubungan antara sejarah trauma dan kejadian PAAL serta membandingkan tahap keterukan PAAL antara gigi trauma dan tanpa trauma selepas enam dan 12 bulan rawatan ortodontik. Sampel kajian merupakan gigi insisor tengah maksila daripada 23 subjek (8 lelaki dan 15 wanita berumur 12 hingga 26 tahun) dengan 19 mempunyai trauma (tanpa penyerapan akar gigi) dan 27 tanpa trauma. Rawatan ortodontik dilakukan dengan menggunakan dawai arkus NiTi 0.014”, 0.018” dan 0.018” × 0.025” pada enam bulan pertama. Selepas enam bulan, rawatan ortodontik diteruskan dengan menggunakan dawai arkus keluli tahan karat saiz 0.019” × 0.025” sehingga rawatan ortodontik mencapai satu tahun. PAAL gigi diukur melalui imej tomografi berkomputer pancaran-kon (CBCT) yang diambil sebelum (X0), selepas enam bulan (X6) dan selepas 12 bulan (X12) rawatan ortodontik. Penyerapan akar dikira dengan menolak panjang gigi pada X6 dan X12 dengan panjang gigi pada X0. Kejadian PAAL dalam kumpulan trauma dan tanpa trauma masing-masing adalah 89.5% dan 77.8% (penyerapan akar rendah dari 2 mm) selepas enam bulan rawatan ortodontik. Selepas 12 bulan rawatan ortodontik, semua gigi menunjukkan PAAL. Kejadian PAAL antara gigi trauma dan tanpa trauma tidak menunjukkan perbezaan yang signifikan (p>0.05). Dalam kajian ini, gigi yang mengalami trauma serta tanpa trauma membentuk PAAL pada kadar yang sama selepas enam dan 12 bulan rawatan orthodontik. Oleh itu, adalah selamat untuk melakukan rawatan ortodontik kepada pesakit yang mempunyai sejarah trauma pada gigi

Stability of lactate dehydrogenase, aspartate aminotransferase, alkaline phosphatase and tartrate resistant acid phosphatase in human saliva and gingival crevicular fluid in the presence of protease inhibitor

The stability of aspartate aminotransferase (AST), lactate dehydrogenase (LDH), tartrate resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP) activities from saliva and gingival crevicular fluid (GCF) with and without the addition of protease inhibitor (PI) at room temperature (RT; 25°C), 4°C and -20°C were investigated. AST, LDH, TRAP and ALP activities in saliva and GCF (n=9) with and without the addition of PI were assayed at 0 (control), 12, 24, 48, 72 h, one and two weeks. A paired t-test showed there were a significant differences (p<0.05) between LDH and TRAP activities in saliva, in the presence and without PI at all temperatures. ALP activity exhibited a significant difference in activity (p<0.05) in the presence and without PI at RT while no significant differences were observed at 4ºC and -20ºC. A significant difference (p<0.05) was observed in AST, LDH and TRAP activities (GCF) at RT and 4ºC in the presence and without PI. We conclude that PI is essential for maintaining stable enzyme activities in saliva and GCF

Intrinsic anticarcinogenic effects of Piper sarmentosum ethanolic extract on a human hepatoma cell line

<p>Abstract</p> <p>Background</p> <p><it>Piper sarmentosum</it>, locally known as kaduk is belonging to the family of Piperaceae. It is our interest to evaluate their effect on human hepatoma cell line (HepG2) for the potential of anticarcinogenic activity.</p> <p>Results</p> <p>The anticarcinogenic activity of an ethanolic extract from <it>Piper sarmentosum </it>in HepG2 and non-malignant Chang's liver cell lines has been previously determined using (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) (MTT) assays, where the IC₅₀value was used as a parameter for cytotoxicity. The ethanolic extract that showed anticarcinogenic properties in HepG2 cells had an IC₅₀of 12.5 μg mL^-1, while IC₅₀values in the non-malignant Chang's liver cell line were greater than 30 μg mL^-1. Apoptotic morphological changes in HepG2 cells were observed using an inverted microscope and showed chromatin condensation, cell shrinkage and apoptotic bodies following May-Grunwald-Giemsa's staining. The percentage of apoptotic cells in the overall population (apoptotic index) showed a continuously significant increase (p < 0.05) in 12.5 μg mL^-1ethanolic extract-treated cells at 24, 48 and 72 hours compared to controls (untreated cells). Following acridine orange and ethidium bromide staining, treatment with 10, 12 and 14 μg mL^-1of ethanolic extracts caused typical apoptotic morphological changes in HepG2 cells. Molecular analysis of DNA fragmentation was used to examine intrinsic apoptosis induced by the ethanolic extracts. These results showed a typical intrinsic apoptotic characterisation, which included fragmentation of nuclear DNA in ethanolic extract-treated HepG2 cells. However, the non-malignant Chang's liver cell line produced no DNA fragmentation. In addition, the DNA genome was similarly intact for both the untreated non-malignant Chang's liver and HepG2 cell lines.</p> <p>Conclusion</p> <p>Therefore, our results suggest that the ethanolic extract from <it>P. sarmentosum </it>induced anticarcinogenic activity through an intrinsic apoptosis pathway in HepG2 cells <it>in vitro</it>.</p

In Vitro Chondrogenesis Transformation Study of Mouse Dental Pulp Stem Cells

A major challenge in the application of mesenchymal stem cells in cartilage reconstruction is that whether the cells are able to differentiate into fully mature chondrocytes before grafting. The aim of this study was to isolate mouse dental pulp stem cells (DPSC) and differentiate them into chondrocytes. For this investigation, morphological, molecular, and biochemical analyses for differentiated cells were used. To induce the chondrocyte differentiation, DPSC were cultured in chondrogenic medium (ZenBio, Inc.). Based on morphological analyses using toluidine blue staining, proteoglycan products appear in DPSC after 21 days of chondrocyte induction. Biochemical analyses in differentiated group showed that alkaline phosphatase activity was significantly increased at day 14 as compared to control (P &lt; 0.05). Cell viability analyses during the differentiation to chondrocytes also showed that these cells were viable during differentiation. However, after the 14th day of differentiation, there was a significant decrease (P &lt; 0.05) in the viability proportion among differentiated cells as compared to the control cells. In RT-PCR molecular analyses, mouse DPSC expressed Cd146 and Cd166 which indicated that these cells belong to mesenchymal stem cells. Coll I and Coll II markers showed high expression after 14 and 21 days, respectively. In conclusion, this study showed that DPSC successfully differentiated into chondrocytes

In Vitro

A major challenge in the application of mesenchymal stem cells in cartilage reconstruction is that whether the cells are able to differentiate into fully mature chondrocytes before grafting. The aim of this study was to isolate mouse dental pulp stem cells (DPSC) and differentiate them into chondrocytes. For this investigation, morphological, molecular, and biochemical analyses for differentiated cells were used. To induce the chondrocyte differentiation, DPSC were cultured in chondrogenic medium (Zen-Bio, Inc.). Based on morphological analyses using toluidine blue staining, proteoglycan products appear in DPSC after 21 days of chondrocyte induction. Biochemical analyses in differentiated group showed that alkaline phosphatase activity was significantly increased at day 14 as compared to control (P<0.05). Cell viability analyses during the differentiation to chondrocytes also showed that these cells were viable during differentiation. However, after the 14th day of differentiation, there was a significant decrease (P<0.05) in the viability proportion among differentiated cells as compared to the control cells. In RT-PCR molecular analyses, mouse DPSC expressed Cd146 and Cd166 which indicated that these cells belong to mesenchymal stem cells. Coll I and Coll II markers showed high expression after 14 and 21 days, respectively. In conclusion, this study showed that DPSC successfully differentiated into chondrocytes