Long-term Follow-up of Complicated Crown Fracture With Fragment Reattachment: Two Case Reports.

Two cases of complicated crown fracture of the maxillary incisors were restored using the fragment reattachment technique. Root canal treatment was performed, and the fractured fragment was bonded to the tooth structure using a dentin adhesive system and a flowable composite resin, followed by the insertion of a fiber post using dual-cured resin cement. Reattached fragments have shown reliable prognosis without inflammatory signs around bonded junctions after long-term follow-up.

Structural characterization of rice starch in rice cake modified by Thermus scotoductus 4-alpha-glucanotransferase (TS alpha GTase).

Rice cake was produced with a thermostable 4-alpha-glucanotransferase from Thermus scotoductus (TS alpha GTase). Starch molecular fine structure, texture, and retrogradation for the enzymatically prepared rice cake were investigated and compared to those for control rice cake. The amylose content in TS alpha GTase-treated rice cakes decreased, whereas branched and linear malto-oligosaccharides ranging from maltose to maltoheptaose increased slightly. The average molecular weight of the enzyme-treated rice starch in rice cake decreased as amylopectin macromolecules were cleaved and reorganized into small amylopectin clusters. The number of shorter side chains (degree of polymerization [DP] < 9) increased, whereas the number of longer side chains (DP > 10) decreased through the disproportionation reaction of TS alpha GTase. After 24 h of storage at 4 degrees C, the enzyme-treated samples demonstrated significantly lower melting enthalpy of retrograded starch (0.4 mJ/mg) compared to that of the control (1.4 mJ/mg). The results indicated that TS alpha GTase treatment effectively inhibited starch retrogradation in rice cakes. It is suggested that the reduction of amylose content, the rearrangement of amylopectin, and the production of malto-oligosaccharides caused by TS alpha GTase treatment are responsible for the ineffective molecular reassociation of rice starch in rice cake.

Congenital disorder of glycosylation Ic in patients of Indian origin.

Congenital disorder of glycosylation type Ic (CDG-Ic) is caused by mutations in ALG6, encoding an alpha 1,3-glucosyltransferase. The most frequent mutation found in this gene (C998T resulting in an A333V substitution) has until now been found only in patients of European origin. Here we describe the first occurrence of this CDG-Ic mutation in patients of Indian origin. Of three Indian patients described in this study, patient 1 was homozygous and patient 2 heterozygous for the A333V mutation. In patient 2 we also found a new mutation, IVS3+2_3insT, just 3bp away from the previously described IVS3+5G>A substitution; both mutations resulted in exon 3 skipping. We screened a panel of >350 genomic DNA samples from an ethnically diverse American population to determine the frequency of the A333V mutation. None of the samples carried this mutation, indicating the frequency of patients carrying this homozygous mutation should be <1 in 5x10(5). The discovery of the common CDG-Ic mutation A333V in an Indian population raises questions as to its ethnic origin.

Mammalian glycosyltransferase expression allows sialoglycoprotein production by baculovirus-infected insect cells.

The baculovirus-insect cell expression system is widely used to produce recombinant mammalian glycoproteins, but the glycosylated end products are rarely authentic. This is because insect cells are typically unable to produce glycoprotein glycans containing terminal sialic acid residues. In this study, we examined the influence of two mammalian glycosyltransferases on N-glycoprotein sialylation by the baculovirus-insect cell system. This was accomplished by using a novel baculovirus vector designed to express a mammalian alpha2,6-sialyltransferase early in infection and a new insect cell line stably transformed to constitutively express a mammalian beta1,4-galactosyltransferase. Various biochemical assays showed that a foreign glycoprotein was sialylated by this virus-host combination, but not by a control virus-host combination, which lacked the mammalian glycosyltransferase genes. Thus, this study demonstrates that the baculovirus-insect cell expression system can be metabolically engineered for N-glycoprotein sialylation by the addition of two mammalian glycosyltransferase genes.