Expression analysis of notch signaling pathway molecules in SHED cultured in keratinocyte growth medium

Aim: To detect the expression of molecules associated with Notch signaling pathway in stem cells from human exfoliated deciduous teeth (SHED) cultured in specific differentiation medium, namely, keratinocyte growth medium (KGM). Methods: RNA was extracted from SHED harvested on day 1, 3 and 7. RNA was reverse-transcribed to obtain the cDNA and then proceeded with PCR using specific primers for the Notch signaling pathway molecules (Notch1, Jagged-1, Jagged-2 and, Hes1) as well as stem cell marker (Nanog). PCR products were electrophoresed on a 2% agarose gel and stained with SYBR green. Results: Notch-1 was highly expressed in SHED cultured in KGM and showed increase in density as the days progressed, while Jagged-1 showed a decrease. Jagged-2 on the other hand, showed a slight increase on day 3 followed by a decrease on day 7. However, Hes-1 was not expressed in SHED cultured in KGM. Nanog showed expression only on day 3 and gradually increased in expression on day 7. Conclusions: Notch signaling pathway associated molecules; Notch-1, Jagged-1, Jagged-2, and stem cell marker Nanog are expressed in SHED cultured in KGM which may be involved in the differentiation into epithelial-like cells in human dental pulp tissues. Keywords: receptors, notch; gene expression; stem cells; tooth, deciduous; culture media

E-content module for chemistry Massive Open Online Course (MOOC): development and students’ perceptions

This study aimed to develop an e-content module for Chemistry Massive Open Online Course (MOOC). It examined the validity, reliability and student’s perceptions on content, usability, design and effectiveness of the module. This is a design and development study and the e-content module was developed based on ADDIE instructional design model. Collaborative learning, Connectivism theory and Interaction Equivalency Theorem are the pillars for the module development. The content validity of the module was estimated by three experts using content validity evaluation form. Questionnaires were distributed to the students in order to determine the reliability (n = 23) and students’ perceptions on the module (n = 129). Findings revealed the e-content module has high content validity (CVI = 1.00) and good reliability index (α = 0.94). The mean scores for students’ perception on module content (M = 3.66, SD = 0.55), usability (M = 3.43, SD = 0.56), design (M = 3.41, SD = 0.59) and effectiveness (M = 3.47, SD = 0.56) constructs were high. This e-content module in Chemistry MOOC is hoped to be a good and useful online resource for both lecturers and students in the teaching and learning of Chemistry in higher education institutionPeer Reviewe

E-content module for Chemistry Massive Open Online Course (MOOC): Development and students’ perceptions

This study aimed to develop an e-content module for Chemistry Massive Open Online Course (MOOC). It examined the validity, reliability and student’s perceptions on content, usability, design and effectiveness of the module. This is a design and development study and the e-content module was developed based on ADDIE instructional design model. Collaborative learning, Connectivism theory and Interaction Equivalency Theorem are the pillars for the module development. The content validity of the module was estimated by three experts using content validity evaluation form. Questionnaires were distributed to the students in order to determine the reliability (n = 23) and students’ perceptions on the module (n = 129). Findings revealed the e-content module has high content validity (CVI = 1.00) and good reliability index (α = 0.94). The mean scores for students’ perception on module content (M = 3.66, SD = 0.55), usability (M = 3.43, SD = 0.56), design (M = 3.41, SD = 0.59) and effectiveness (M = 3.47, SD = 0.56) constructs were high. This e-content module in Chemistry MOOC is hoped to be a good and useful online resource for both students and lecturers in the teaching and learning of Chemistry in higher education institution

Expression analysis of Notch signaling pathway molecules in SHED cultured in keratinocyte growth medium

detect the expression of molecules associated with Notch signaling pathway in stem cells from human exfoliated deciduous teeth (SHED) cultured in specific differentiation medium, namely, keratinocyte growth medium (KGM). Methods: RNA was extracted from SHED harvested on day 1, 3 and 7. RNA was reverse-transcribed to obtain the cDNA and then proceeded with PCR using specific primers for the Notch signaling pathway molecules (Notch1, Jagged-1, Jagged-2 and, Hes1) as well as stem cell marker (Nanog). PCR products were electrophoresed on a 2% agarose gel and stained with SYBR green. Results: Notch-1 was highly expressed in SHED cultured in KGM and showed increase in density as the days progressed, while Jagged-1 showed a decrease. Jagged-2 on the other hand, showed a slight increase on day 3 followed by a decrease on day 7. However, Hes-1 was not expressed in SHED cultured in KGM. Nanog showed expression only on day 3 and gradually increased in expression on day 7. Conclusions: Notch signaling pathway associated molecules; Notch-1, Jagged-1, Jagged-2, and stem cell marker Nanog are expressed in SHED cultured in KGM which may be involved in the differentiation into epithelial-like cells in human dental pulp tissue

Expression of GBS virulence genes in high vaginal swabs of symptomatic pregnant women at Hospital Tengku Ampuan Afzan, Kuantan

During pregnancy, group B streptococcus (GBS) colonization is known as one of the risk factors for preterm birth and consequently neonatal infections. Previous in-vitro experiments using human cells and in vivo animal models have portrayed the important roles of these virulence factors including hemolytic pigment (CylE), hyaluronidase (HylB), serine-rich protein (Srr) and bacterial surface adhesion of GBS (BsaB) in mediating GBS colonization and intrauterine ascending infection, leading to preterm delivery. The aim of this study is to investigate the association between mRNA expression of women and preterm delivery. GBS isolates were obtained from high vaginal swabs of pregnant women(n=40) with gestational age less than 37 weeks and symptoms including preterm labour, preterm premature rupture of membrane (pPROM), vaginal discharge and vaginal bleeding. Socio-demographic details, obstetric history and delivery outcomes of these women were also enquired. RNA was extracted from these GBS isolates and RT-qPCR was performed to determine the relative mRNA expression of GBS virulence genes including CylE,HylB, Srr and BsaB. Socio-demographic details and obstetric history were not found to be associated with the delivery outcomes of these women. Women with preterm labour and pPROM who delivered prematurely were demonstrated with higher expression of HylB and Cyl Egenes, in comparison to women with term delivery. The expression of Srr and BsaB genes were both similar between symptomatic pregnant women who had term and preterm delivery. Theseresultssuggestthatfollowingvaginalcolonization,bothCylEandHylBgenespossiblycontributetointrauterineascendinginfectionandinflammation,leading to preterm delivery in humans.Thus, hemolytic pigment and hyaluronidase may be targeted for exploratory and pre-clinical stages of vaccine development, which is a good alternative to intrapartum antibiotic prophylaxis in order to prevent neonatal GBS infections

Expression of GBS virulence genes in high vaginal swabs of symptomatic pregnant women at Hospital Tengku Ampuan Afzan, Kuantan

During pregnancy, group B streptococcus (GBS) colonization is known as one of the risk factors for preterm birth and consequently neonatal infections. Previous in-vitro experiments using human cells and in vivo animal models have portrayed the important roles of these virulence factors including hemolytic pigment (CylE), hyaluronidase (HylB), serine-rich protein (Srr) and bacterial surface adhesion of GBS (BsaB) in mediating GBS colonization and intrauterine ascending infection, leading to preterm delivery. The aim of this study is to investigate the association between mRNA expression of women and preterm delivery. GBS isolates were obtained from high vaginal swabs of pregnant women(n=40) with gestational age less than 37 weeks and symptoms including preterm labour, preterm premature rupture of membrane (pPROM), vaginal discharge and vaginal bleeding. Socio-demographic details, obstetric history and delivery outcomes of these women were also enquired. RNA was extracted from these GBS isolates and RT-qPCR was performed to determine the relative mRNA expression of GBS virulence genes including CylE,HylB, Srr and BsaB. Socio-demographic details and obstetric history were not found to be associated with the delivery outcomes of these women. Women with preterm labour and pPROM who delivered prematurely were demonstrated with higher expression of HylB and Cyl Egenes, in comparison to women with term delivery. The expression of Srr and BsaB genes were both similar between symptomatic pregnant women who had term and preterm delivery. Theseresultssuggestthatfollowingvaginalcolonization,bothCylEandHylBgenespossiblycontributetointrauterineascendinginfectionandinflammation,leading to preterm delivery in humans.Thus, hemolytic pigment and hyaluronidase may be targeted for exploratory and pre-clinical stages of vaccine development, which is a good alternative to intrapartum antibiotic prophylaxis in order to prevent neonatal GBS infections