Substrate specificity of the amino acid transporter PAT1.

The proton coupled amino acid transporter PAT1 expressed in intestine, brain, and other organs accepts L- and D-proline, glycine, and L-alanine but also pharmaceutically active amino acid derivatives such as 3-amino-1-propanesulfonic acid, L-azetidine-2-carboxylic acid, and cis-4-hydroxy-D-proline as substrates. We systematically analyzed the structural requirements for PAT1 substrates by testing 87 amino acids, proline homologs, indoles, and derivatives. Affinity data and effects on membrane potential were determined using Caco-2 cells. For aliphatic amino acids, a blocked carboxyl group, the distance between amino and carboxyl group, and the position of the hydroxyl group are affinity limiting factors. Methylation of the amino group enhances substrate affinity. Hetero atoms in the proline template are well tolerated. Aromatic alpha-amino acids display low affinity. PAT1 interacts strongly with heterocyclic aromatic acids containing an indole scaffold. The structural requirements of PAT1 substrates elucidated in this study will be useful for the development of prodrugs.

The craniofacial skeleton in anencephalic human fetuses. II. Calvarium.

A detailed study of the calvarium of twelve anencephalic and four normal human fetuses 26 to 40 weeks gestational age using gross dissection, alizarin red S staining, silver nitrate radiography and histology revealed dramatic alterations in the presence, form, location and relationship of the individual bones. In the larger dorsal cranial defects the interparietal portions of the occipital bone were relocated anteriorly to approximate the frontal bone. The occipital components were rotated anterolaterally and inferiorly with lack of fusion of the chondrocranium posterior to the foramen magnum. The squamae of the frontal bone were collapsed horizontally and reduced in size to lie peripheral to the anterior cranial fossa forming most of the orbital roofs. In anencephaly the bones derived from the chondrocranium were not as severely affected morphologically as those derived from the neurocranium. The sutures were narrow and smooth instead of wide and serrated as in the normally developing calvarium. In general the degree of maldevelopment was proportional to the extent of the dorsal cranial defect in anencephaly.

The craniofacial skeleton in anencephalic human fetuses. III. Facial skeleton.

A sample of 12 anencephalic fetuses with gestational ages ranging from 26 to 40 weeks and exhibiting varying degrees of severity of the dorsal cranial defect was compared to three normal fetuses of comparable gestational ages with regard to the morphology and positional relationships of the maxillofacial skeletal complex. Gross dissection, alizarin red S staining, radiographs, cephalometric tracings, and histologic techniques were utilized. It was found that some facial bones were severely affected in morphology, size, spatial and angular relationships. The manner in which these were altered suggests that their morphogenesis is an adaptation to the primary defect of the neurocranium.

The craniofacial skeleton in anencephalic human fetuses. I. Cranial floor.

Twelve anencephalic and four normal fetuses 26 to 40 weeks gestational age were compared by anatomic, radiographic and histologic methods in order to gain information concerning morphogenesis. In the anencephalics, alterations located within the body of the sphenoid bone led to a reduced cranial floor angle and a more vertical clivus. The reduced lateral extension of the lesser and greater wings of the sphenoid constricted the anterior and middle cranial fossae respectively. The posterior cranial fossa tended to have an increased transverse dimension related to the supraoccipital and exoccipital bone orientation. The increased anterior and inferior position of the lateral end of the petrous temporal ridge was positively correlated with the degree of dorsal schisis in the anencephalics. Alterations in the size, form, or duration of the neural functional matrix are suggested as the cause of changes in the cranial floor.