ABSTRACT
Perivitelline fluid, extracted from the fertilized eggs of horseshoe crabs, has been reported to play a vital role in supporting embryogenesis as well as cell proliferation. The present study aims to evaluate the effect of PVF on the expression of COL1A1 in human dental pulp stem cells (DPSCs). The cells were grouped into two; untreated (control) and treated with a single dose of PVF (0.019 mg/ml). Gene expression was quantified for COL1A1 on day 1, 3 and 7 using reverse transcriptase PCR. The expression of COL1A1 on day 3 of treated group with PVF was the highest though there was a decline of COL1A1 expression on day 7. Mann Whitney test was utilized to determine the significance of COL1A1 expression between treated and untreated groups. Significant difference in the expression of COL1A1 was observed between the treated and untreated groups on day 3 though there was no significance in the expression on day 7. The present study indicates that PVF may have the potential to increase cell proliferation in human DPSCs.
Subject(s)
Dental Pulp , Horseshoe Crabs , Stem CellsABSTRACT
Perivitelline fluid (PVF) of the horseshoe crab embryo has been reported to possess an important role during embryogenesis by promoting cell proliferation. This study aims to evaluate the effect of PVF on the expression of cell cycle regulatory genes from human dental pulp stem cells (DPSCs) between different cell passages viz. 4, 5, 6. The cells were treated with a single dose of PVF (26.89 mg/ml) PVF. Gene expression was quantified for CDKNA2A, PTEN, MDM2 and TP53 genes using reverse transcriptase PCR. CDKN2A and MDM2 expression for treated and untreated DPSCs, expressed a similar pattern of expression. The higher expression of CDKN2A showed that the treatment increased cell proliferation and prevented cell senescence. DPSCs with PVF treatment showed increased expression of MDM2 at passage 4 and drastically declined expression at passage 5 and slightly increased at passage 6. TP53 expression of DPSCs treated group showed a higher expression compared to untreated group. On the other hand, the expression of PTEN in DPSCs treated group started to increase from passage 5 to 6. However, on the whole, the PTEN expression was higher than the untreated group in all the passages studied here. The results showed that PVF could enhance cell cycle regulatory gene expression in DPSCs as indicated by the higher expression of all the genes considered in this study at different cell passages in the treated group compared to the untreated group. Mann Whitney test was utilized to determine the significance of cell cycle regulatory genes expression between treated and untreated group. Significant difference in expression of genes between the treated and untreated groups were found at all passages except for CDKN2A gene whereby, its expression was not significantly different at passage 5 though it did express slightly higher in PVF treated DPSCs.
ABSTRACT
The trilobite larvae of C. rotundicauda were tested to determine their colour preference and light sensitivity until their first moulting (25 days post hatching) under laboratory conditions. Maximum congregation size of the trilobite larvae was found in the white zone respectively where (n= 12) followed by yellow (n= 8) and orange (n= 8), which showed the larval preference for lighter zones. Morisita’s index calculation showed a clumped/aggregated distribution (yellow, blue, orange and white) and uniform/hyper dispersed distribution (green, red and black) for various colours tested. Trilobite larvae showed least preference for brighter regions while tested in the experiment [black; (n=4) and red; (n=5)]. Experiments done to determine the light sensitivity of trilobite larvae showed that the larvae had more preference towards ultraviolet lights. The maximum congregation size of 38.8 and 40.7% of the larvae was encountered under ultraviolet light, when the light sources were kept horizontal and vertical, respectively. Overall, results suggested that the trilobite larvae of C. rotundicauda, preferred light source of shorter wavelengths (UV light) and colours of lighter zone (white, yellow, orange), which might be due to their adaptation to their natural habitat for predator avoidance, prey selection and water quality.
Subject(s)
Animals , Color , Horseshoe Crabs/physiology , Larva/physiology , Photophobia , Ultraviolet RaysABSTRACT
Purpose:Study the changes of mitochondria membrane potential of HL 60 cell induced by horseshoe crab hemocyte peptides. Methods:HL 60 cells were treated with horseshoe crab hemocyte peptides 0,12.5,25,50,100 ?g/ml for 2,4,6,8,10,12,14h. HL 60 cells were stained with Rhodamine123 before mitochondria membrane potential was detected. Change of mitochondria membrane potential of HL 60 cell was analyzed by flow cytometry. Results:Mitochondria membrane potential of HL 60 cells was stable from 0 to 14h in control groups,however mitochondria membrane potential was decreased when HL 60 cells were treated with horseshoe crab hemocyte peptides 12.5～100 ?g/ml for 0 to 14h.Conclusions:Decrease of mitochondria membrane potential was dependant on concentrations of peptides and time of peptides treatment, it was relative between horseshoe crab hemocyte peptides inducing HL 60 cells apoptosis and decrease of mitochondria membrane potential.
ABSTRACT
The binding affinities of some ligands towards the sialic acid-specific lectin carcinoscorpin, from hemolymph of the horseshoe crab Carcinoscorpius rotundacauda have been determined by protein fluorescence quenching in presence of ligands. Among the ligands studied, the disaccharide O-(N-acetylneuraminyl)-(2→6)-2-acetamido-2-deoxy-Dgalactitol has the highest Ka(l.15 × 106 M–1) for carcinoscorpin. Studies on the effect of pH on Ka values of disaccharide suggests the possible involvement of amino acid residues having pKa values around 6.0 and 9.0 in the binding activity of carcinoscorpin. There were distinct changes in the accessibility of the fluorescent tryptophan residues of carcinoscorpin by ligand-binding as checked through potassium iodide quenching.