Delivery of active agents from chewing gum for improved remineralization.

Most surrogate measures of caries were developed to test products containing fluoride, typically at relatively high and closely controlled oral concentrations. However, since the primary mechanism for the remineralization of early enamel caries lesions by chewing gum is through stimulation of saliva, delivering Ca and Pi to the demineralized enamel lesion, established methods may lack the sensitivity to detect the additional benefit of an active agent without the strong remineralizing potential of fluoride. Issues related to the release of active agents from the gum matrix, dilution in the saliva, and limited oral retention time, along with taste, safety, regulatory, and cost concerns, impose further limitations. This paper reviews the efficacy of some active agents used in chewing gum for improved remineralization and includes results from in situ testing of calcium-containing gums, including calcium lactate, tetracalcium phosphate/dicalcium phosphate anhydrous, calcium citrate/encapsulated phosphate, and a calcium lactate/sodium phosphate blend. Despite promising in vitro data from these agents, they did not provide consistently superior results from in situ testing. There is a need to develop better predictive in vitro models for chewing gum, as well as improved sensitivity of in situ models to discriminate relatively small amounts of remineralization against a background of high biological variability.

Muscarinic receptor- and phorbol ester-stimulated phosphorylation of protein kinase C substrates in adult and neonatal cortical slices.

The neuron-specific protein B-50 (GAP-43) is a major presynaptic substrate for protein kinase C (PKC). Phosphorylation of B-50 by PKC at serine-41 is functionally related to signal transduction in association with process outgrowth and neurotransmitter release. Thus, it is important to characterize the factors which modulate phosphorylation of B-50 by PKC. Phosphoinositide (PI)-coupled muscarinic acetylcholine receptor (mAchR) activation would be expected to increase PKC activity through production of the second messenger, diacylglycerol. To test the hypothesis that activation of mAchR also increases phosphorylation of B-50, protein phosphorylation has been examined in cerebral cortical slices in response to the cholinergic agonist, carbachol (Cch) in comparison to the phorbol ester, 4beta-phorbol 12, 13-dibutyrate (PDB), a known activator of PKC. At short times of incubation with 1 mM Cch, a concentration which maximally activates PI metabolism, increased phosphorylation of a group of synaptosomal proteins, including B-50 and myristoylated, alanine-rich C kinase substrate (MARCKS), was observed. This increase was approximately half of that obtained in response to 1 microM PDB. Differing patterns of protein phosphorylation were observed in neonatal and adult slices: neonatal samples contained more MARCKS and a PKC substrate with a Mr of 46 kDa. Phosphorylation of B-50 and MARCKS was sensitive to Cch in both cases. Immunoblotting demonstrated less m1 acetylcholine receptor (the predominant mAchR subtype coupled to PI metabolism in the cortex) in neonatal, as compared to adult, synaptosomal fractions. These results are consistent with a coupling between mAchR-stimulated PI metabolism and PKC-mediated protein phosphorylation that is developmentally regulated.

Pondering the procephalon: the segmental origin of the labrum.

With accumulating evidence for the appendicular nature of the labrum, the question of its actual segmental origin remains. Two existing insect head segmentation models, the linear and S-models, are reviewed, and a new model introduced. The L-/Bent-Y model proposes that the labrum is a fusion of the appendage endites of the intercalary segment and that the stomodeum is tightly integrated into this segment. This model appears to explain a wider variety of insect head segmentation phenomena. Embryological, histological, neurological and molecular evidence supporting the new model is reviewed.

Homeotic evidence for the appendicular origin of the labrum in Tribolium castaneum.

The ontogeny of the insect labrum, or upper lip, has been debated for nearly a century. Recent molecular data suggest a segmental appendage origin of this structure. Here we report the first arthropod mutation associated with a homeotic transformation of the labrum. Antennagalea-5 (Ag(5)) transforms both antennal and labral structures to resemble those of gnathal appendages in Tribolium castaneum. This labral transformation suggests that the labrum is a fused structure composed of two pairs of appendage endites, and is serially homologous to the gnathal appendages.

Genetic analysis of the homeotic gene complex (HOM-C) in the beetle Tribolium castaneum.

Our laboratories have undertaken both genetic and molecular studies of the homeotic gene complex (HOM-C) of the beetle Tribolium castaneum, and this paper discusses results from our genetic analyses. We describe here the adult phenotypes and complementation behavior of over 50 new mutations. Many of these homeotic phenotypes resemble those of Drosophila melanogaster, but few precisely parallel the segmental transformations seen in this fly. Analysis of putative loss-of-function mutations affecting the head and thorax suggests that the maxillopedia and Cephalothorax genes most closely resemble proboscipedia and Sex combs reduced of Drosophila. In the abdomen, putative loss-of-function alleles of Abdominal affect a domain corresponding to those of the combined abdominal-A and Abdominal-B genes of Drosophila. In contrast to the situation in flies, Abdominal loss-of-function variants in Tribolium cause anteriorward transformations in A3-A5a, but posteriorward transformations in A5p-A7. The implications of the differences in developmental strategies evolved in Tribolium vs Drosophila are discussed.

Chromosome extraction and revision of linkage group 2 in Tribolium castaneum.

We used a balancer chromosome to recover ethylmethanesulfonate-induced recessive mutations in a targeted region of the genome of the red flour beetle (Tribolium castaneum) by the technique of chromosome extraction. The experiments reported herein constitute the first successful application of this powerful technique in the order Coleoptera. Using the balancer chromosome maxillopedia-Dachs3 (mxpDch-3), we recovered seven recessive visible variants representing seven distinct loci and several dozen recessive lethal variants representing at least five distinct loci after screening 1,607 EMS-mutagenized chromosomes. A subset of the mxpDch-3-extracted mutations were positioned on the map of the second linkage group by a series of two-, three-, and four-point crosses. The orientation of the homeotic gene complex (HOM-C) on this linkage group was also determined. With the advent of better and more varied balancer chromosomes and the concomitant improvement of chromosome extraction procedures for genetic analysis of T. castaneum, saturation mutagenesis of targeted regions of the genome is now feasible in this species.