Small Ubiquitin-Like Modifier Protein 3 Enhances the Solubilization of Human Bone Morphogenetic Protein 2 in E. coli.

Small ubiquitin-like modifier (SUMO) fusion technology is widely used in the production of heterologous proteins from prokaryotic system to aid in protein solubilization and refolding. Due to an extensive clinical application of human bone morphogenetic protein 2 (hBMP2) in bone augmentation, total RNA was isolated from human gingival tissue and mature gene was amplified through RT-PCR, cloned (pET21a), sequence analyzed, and submitted to GenBank (Accession no. KF250425). To obtain soluble expression, SUMO3 was tagged at the N-terminus of hBMP2 gene (pET21a/SUMO3-hBMP2), transferred in BL21 codon+, and ~ 40% soluble expression was obtained on induction with IPTG. The dimerized hBMP2 was confirmed with Western blot, native PAGE analysis, and purified by fast protein liquid chromatography with 0.5 M NaCl elution. The cleavage of SUMO3 tag from hBMP2 converted it to an insoluble form. Computational 3D structural analysis of the SUMO3-hBMP2 was performed and optimized by molecular dynamic simulation. Protein-protein interaction of SUMO3-hBMP2 with BMP2 receptor was carried out using HADDOCK and inferred stable interaction. The alkaline phosphatase assay of SUMO3-hBMP2 on C2C12 cells showed maximum 200-ng/ml dose-dependent activity. We conclude that SUMO3-tagged hBMP2 is more suited for generation of soluble form of the protein and addition of SUMO3 tag does not affect the functional activity of hBMP2.

Haematological parameters and recurrent aphthous stomatitis.

OBJECTIVE: To find out the relationship between recurrent aphthous stomatitis (RAS) with deficiencies of haemoglobin, haematocrit, serum vitamin B12, serum Ferritin and red blood cells (RBC) Folate level. STUDY DESIGN: An analytical cross-sectional study. PLACE AND DURATION OF STUDY: Department of Oral Health Sciences, Shaikh Zayed Federal Postgraduate Medical Complex, Lahore, from February to July 2008. METHODOLOGY: Sixty consecutive subjects with active RAS were taken as the aphthous group; 60 age and gender matched subjects without RAS were as the Non-Aphthous group. Five milliliter blood was taken from both groups to evaluate the levels of serum B12, and RBC Folate through radio immuno assay and serum ferritin with enzyme linked immuno-sorbent assay tests. Complete blood count was carried out to determine the level of haemoglobin and haematocrit in both groups. Proportion of subjects with lower values was compared using 2 text of proportions with significance at p < 0.05. RESULTS: Serum Ferritin (p = 0.001), haematocrit (p < 0.001), RBC Folate (p < 0.001) and serum B12 (p < 0.001) were significantly lower in the RAS group. Combined deficiency state (haemoglobin, serum Ferritin, haematocrit, RBC Folate and serum B12) was identified in 13% (n = 8) RAS patients. CONCLUSION: Frequency of haematinic deficiencies was high in RAS patients. Serum B12 and RBC Folate were significantly low in aphthous group.

Outcome of labour in nullipara at term with unengaged vertex.

BACKGROUND: Primigravidas with unengaged foetal head at term should be regarded as high risk cases. It is seen that nuglliparous women with the floating foetal head demonstrate higher rates of caesarean section than those with dipping or engaged head in early labour. These cases should be regarded as high risk and identified early and should be referred to a tertiary care centre where good facilities for operative delivery are available. The present study was undertaken to determine the outcome of labour in nulliparous women with unengaged head and compare it with those who present with engaged head at term as a case control study. The objective of the study was to enumerate the outcome of labour in nulliparous women presenting with unengaged head at term and compare it with those who present with engaged head in early active labour. METHODS: It was a case control study, completed in 11 months, conducted in Department of Obstetrics and Gynaecology, Sir Ganga Ram Hospital, Lahore. 300 women were selected by convenience sampling. 150 nulliparous women who presented with unengaged head in early active labour were taken as cases, out which 2 women left against medical advice with their medical records so data was missing leaving 148 patients in this group. While 150 nulliparous women who presented with engaged head were taken as controls. RESULTS: C-section rates was significantly found to be more in unengaged group being 16.89%, compared with 5.33% in engaged group (p = 0.000). Most of C-sections were carried out due to failed progress of labour (48%). Patients with unengaged foetal head had significantly lower APGAR scores at 1 (p < 0.002) and 5 min (0.003) and higher mean birth weights (p = 0.002).Cases also had significantly longer 1st (p = 0.0001) and 2nd stage (p = 0.004) of labour. CONCLUSION: Engaged vertex at the onset of active labour is associated with a lower risk of caesarean delivery in nulliparous women. Patients with unengaged vertex are at higher risk for caesarean delivery due to arrest disorders. In addition to the higher caesarean rate patients with unengaged vertex are at risk of having lower foetal APGAR scores and higher mean birth weights of newborns.