Arylquins target vimentin to trigger Par-4 secretion for tumor cell apoptosis.

The tumor suppressor protein prostate apoptosis response-4 (Par-4), which is secreted by normal cells, selectively induces apoptosis in cancer cells. We identified a 3-arylquinoline derivative, designated Arylquin 1, as a potent Par-4 secretagogue in cell cultures and mice. Mechanistically, Arylquin 1 binds vimentin, displaces Par-4 from vimentin for secretion and triggers the efficient paracrine apoptosis of diverse cancer cells. Thus, targeting vimentin with Par-4 secretagogues efficiently induces paracrine apoptosis of tumor cells.

Localization of alpha/beta and gamma/delta T lymphocytes in Cryptosporidium parvum-infected tissues in naive and immune calves.

The nature of the host's T-lymphocyte population within the intestinal villi following Cryptosporidium parvum infection was characterized with a bovine model of cryptosporidiosis. In naive animals, infection with C. parvum resulted in substantial increases in the numbers of alpha/beta T cells, both CD4+ (150%) and CD8+ (60%), and of gamma/delta T cells (70%) present within the intestinal villi of the infected ileum. In immune animals, the host T-lymphocyte response to a challenge infection with C. parvum was restricted to alpha/beta T cells. The number of CD4+ T cells within the Peyer's patch of the ileum increased dramatically; however, there was little change in the number or localization of CD4+ T cells within the intestinal villi. In contrast, the number of CD8+ T cells within the intestinal villi increased following a challenge infection. In addition, the CD8+ T cells were found to be intimately associated with the epithelial cells of the intestinal villi. The precise correlation between the accumulation of CD8+ T cells and the normal site of parasite development suggests an important role for CD8+ T cells in the immune animal.

Expression of the IV (reversed and/or heterotaxic) phenotype in SWV mice.

Approximately 50% of iv/iv mice have situs inversus (mirror image reversal of viscera) and 40% have heterotaxia (anomalous arrangement of viscera). The occurrence of heterotaxia is independent of situs. Using the cross-intercross breeding system to put the iv gene on the SWV background, an occasional presumed iv/+ mouse was found that had an IV (situs inversus and/or heterotaxic) phenotype. Testcrosses of these reversed animals indicated an iv/+ genotype. Since iv is linked tightly to Igh-C on chromosome 12, we inferred the genotype with a polymorphism of Igh-C demonstrated using the polymerase chain reaction (PCR). This confirmed them to be iv/+. The expression of the IV phenotype in animals heterozygous for the iv gene may be due to an interaction of iv with an autosomal recessive gene found in SWV. We have not found the IV phenotype in heterozygous iv/+ mice following placement of the iv gene on six other inbred strains. Rarely, we also found that presumed SWV +/+ mice had the IV phenotype. Test matings of these phenodeviants, corroborated by PCR, have confirmed them to be +/+. Although the phenotypes of the affected SWV +/+ and iv/+ mice resembled those found in iv/iv mice, the occurrence of situs inversus and heterotaxia were not independent of each other, and most of the SWV mice with the IV phenotype had heterotaxia with situs solitus. This infrequent dominant expression of the iv gene has so far only been seen when iv is on the SWV background. These findings are consistent with the idea that this phenomenon is due to the interaction of the iv gene with another gene found so far only in the SWV strain.

31P magnetic resonance spectroscopy in Alzheimer's and Pick's disease.

Previous studies have suggested a defect in phosphorus metabolism in Alzheimer's disease (AD) gray matter. We have studied phosphorus metabolites in both gray and white matter in autopsy specimens of nine subjects with late-stage AD, three with Pick's disease and seven age-matched controls. Phosphorus metabolites sugar phosphate (SU), phosphomonoester (PME), phosphodiester (PD), and inorganic phosphate (PI) were quantified as mole percentages in regional neocortical specimens using nuclear magnetic resonance spectroscopy. Senile plaque (SP) and neurofibrillary tangle (NFT) counts were determined in adjacent cortical sections. In the inferior parietal lobule gray and white matter, mole percentage normalized PME, and PD were significantly greater than control values in both AD and Pick's disease. A significant correlation was found between PD and NFT in AD parietal gray matter. Our data indicates that phosphorus metabolite alterations are present in two cortical degenerative diseases and are not likely to be specific for AD.

Situs inversus in the developing mouse: proteins affected by the iv mutation (genocopy) and the teratogen retinoic acid (phenocopy).

To decipher genes that are important in the determination of laterality, we compared two-dimensional protein gels from wild-type C57BL/6J mice and C57BL/6J mice that carried the iv mutation, which confers random determination of visceral situs. To span the time period(s) during which laterality determination occurs, we compared computer-analyzed two-dimensional protein gels from wild-type mouse embryos and iv/iv mouse embryos at 7.5, 8.0, and 8.5 days post-coitum. One polypeptide that was expressed only on day 8.0 of development and only in wild-type embryos represents a particular candidate for determination of laterality. Day 8.5 postcoitum represents the earliest time in murine development that laterality is manifest. Two-dimensional gels were compared from 8.5 day embryos that were C57BL/6J wild-type, C57BL/6J iv/iv, or C57BL/6J wild-type and exposed to the teratogen retinoic acid late on day 7. Reproducible alterations of protein synthesis were observed in both the iv genocopy and retinoic acid phenocopy, yielding abnormal laterality determination. The intersection of these peptide changes identifies a protein likely to play a role in the determination of laterality.

The murine situs inversus viscerum (iv) gene responsible for visceral asymmetry is linked tightly to the Igh-C cluster on chromosome 12.

The iv gene controls left-right determination during murine organogenesis. To map this gene, we analyzed backcross progeny produced by mating (C57BL/6J X MEV/Ty)F1-iv/+heterozygotes to C57BL/6J-iv homozygotes. Hybridization of a murine ecotropic virus probe and several homeotic box gene probes coupled with analysis of dominant visible markers enabled us to exclude the iv locus from much of the mouse genome. Spurred by a recent report that mapped the iv gene to mouse chromosome 12 which was not excluded by our previous work, we used the polymerase chain reaction on our larger cohort to determine that the iv gene is indeed linked tightly to the Igh-C locus on this chromosome: we observed 0/156 recombinants between the iv and Igh-C loci. Combining data from the two studies demonstrates that the murine iv gene is close (1/201 recombinants) to the Igh-C cluster on chromosome 12.

Genetics, chance, and morphogenesis.

We posit that chance plays a major role in the occurrence of many common malformations that cluster in families but recur less frequently than expected for simple Mendelian traits. Once the role of random effects is accepted, the segregation of such malformations may be explained on the basis of Mendelian transmission of a single abnormal gene that predisposes to, but does not always result in, the abnormal phenotype. We apply a stochastic (probabilistic) single-gene model to the occurrence of malformations in mouse and man. The stochastic single-gene model suggests the feasibility of isolating individual genes that determine morphogenesis and sets limits on the precision with which the recurrence of malformations can be predicted.

Isolation and characterization of N-methyl-N'-oxonicotinium ion, a new urinary metabolite of R-(+)-nicotine in the guinea pig.

Biotransformation of both R-(+)-nicotine and R-(+)-N-methyl-nicotinium acetate in male Hartley guinea pigs affords a new quaternary amine metabolite, which was isolated and purified from urine by preparative HPLC. The structural analysis of the metabolite was carried out using UV spectrophotometry, direct thermospray mass spectrometry, and Fourier transform 1H-NMR spectroscopy. The structure of the metabolite was confirmed by synthesis and shown to be a mixture of the cis-1'S,2'R-, and trans-1'R,2'R-diastereomers of N-methyl-N'-oxonicotinium ion, formed in the ratio 1.6:1.0, respectively. S-(-)-Nicotine under similar conditions does not undergo this biotransformation.

Detection of tetrafluoroputrescine in RBCs by fluorine-19 nuclear magnetic resonance spectroscopy.

Fluorine-19 nuclear magnetic resonance spectroscopy is used to detect the in vitro uptake of tetrafluoroputrescine, a novel putrescine analogue, in red blood cells.

Splenic erythropoiesis in polycythemic response of the rat to high-altitude exposure.

Intact rats exposed for 30 days to various levels of simulated altitude from 12,000 (3,658 m) to 20,000 ft (6,096 m) showed a sharp increase in circulating red blood cells in reticulocytes, and in spleen-to-body weight ratios above 15,000 ft (4.572 m). Nucleated erythrocytes in splenic section increased significantly at 18,000 ft (5,486 m), but not at 12,000 ft. Acute splenectomy 1 day before killing sharply reduced the reticulocyte counts at 18,000 and 20,000 ft, but the red cell counts were not reduced at any altitude by the operation. Indeed, at 18,000 ft the splenectomy significantly increased the degree of polycythemia. With altitude exposure the spleen but not the liver or the bone marrow showed an increased 59Fe uptake that was related to the degree of hypoxia. These results suggest that the rat spleen of the present strain carries the full load of the erythropoietic effort in response to a hypoxic stimulus, and that it may exert an inhibitory influence on any extraerythropoietic effort by the bone marrow. In intact rats returned from 18,000 ft to sea level, the reticulocytosis is reversed much more slowly than it is in splenectomized rats, suggesting the presence of a persistent stimulus initiated by hypoxia or a committed pool of reticulocyte precursors.

The susceptibility of inbred strains of hamsters to cadmium-induced embryotoxicity.

Prior-research has involved several aspects of cadmium-induced embryotoxicity in non-inbred hamsters, mice, and rats. This study compares the embryotoxic profile of cadmium in one non-inbred (LVG) and five inbred (CB, LHC, LSH, MHA, PD4) strains of hamsters. A single IV dose (2 mg/kg) of cadmium sulfate was injected into pregnant hamsters early on the 8th gestation day. Fifteenth gestation day fetuses from treated, and control animals were studied for the kinds and incidence of external, internal, and skeletal malformations, as well as the frequency of resorptions. All six hamster strains developed significant resorption rates and external, internal, and skeletal abnormalities, e.g., exencephaly, microphthalmia, cleft lip, cleft palate, renal agenesis, rib fusions, etc. Significant interstrain differences were detected in only three categories of embryonic damage, i.e., resorptions, microphthalmia, and renal agenesis. The data of this study was compared with that of another study in which the same hamster strains were exposed to the metallic teratogen, lead. While the manner by which cadmium damages the mammalian embryo is unknown at this time, several possibilities are presented.

Cadmium induced limb defects in mice: strain associated differences in sensitivity.

Cadmium (CdSO4) was given ip on day 9 at 12 or 24 mumol/kg to pregnant CD-1 (non-inbred) mice. Fetuses showed malformations of the limbs, face, trunk, and tail. There was a statistically significant relationship between the dose of cadmium and the malformation rate. Cadmium (12 mumol/kg ip on day 9) was then given to mice of six inbred strains three of which (A/J, BALB/cJ, and C57BL6J) carry a gene cdm for resistance to cadmium-induced testicular damage, and three strains (AKR/J, CBA/J, and DBA/2J) which do not. Paradoxically, the three strains resistant to cadmium induced testicular damage were significantly more sensitive to its teratogenic effects than were the other three strains. In all inbred strains most malformations involved the limbs. All forelimb defects found in inbred or non-inbred cadmium treated mice were postaxial and indistinguishable from those produced by acetazolamide in mice. The remarkable similarity of the cadmium- and acetazolamide-induced forelimb malformations may be a reflection of the limited number of ways that a rodent forelimb can react to a teratogenic insult. The hindlimb defects were all preaxial.

Carbon-13 nuclear magnetic resonance investigations into the interactions of bisulfite with pyrimidine nucleosides and nucleotides.

Carbon-13 NMR is utilized to demonstrate the attack of bisulfite anion on uridine, 5-fluorouridine, and uridine 5'-monophosphate. The attack produces a pair of diastereomeric adducts similar in structure to those seen in the uracil series. Intensity data from the equilibrium system give an estimate for the individual equilibrium constants. Thymidine and thymidine 5'-monophosphate show no evidence of nucleophilic attack by bisulfite. This evidence indicates that bisulfite addition to nucleosides and nucleotides models the enzymatic methylation of uridine by the enzyme thymidylate synthetase better than the uracil bisulfite system.

Craniomyeloschisis: a spontaneous mutation of the rat.

Craniomyeloschisis (proposed gene symbol cms) was a spontaneous mutation of the rat inherited as an autosomal recessive trait with complete penetrance. Homozygous offspring died at birth with failure of closure of the neural tube caudal to the midbrain. Neural crest derivatives were relatively normal. There were associated severe malformations of the axial skeleton including skull, vertebral column, and ribs. The pattern of malformations was quite uniform. Heterozygotes were anatomically normal. The mutation is extinct.

Random determination of a developmental process: reversal of normal visceral asymmetry in the mouse.

Situs inversus viscerum in the mouse has been shown to be inherited as an autosomal recessive trait (gene symbol iv) with reduced penetrance. It is hypothesized that the normal allele at the iv locus exhibits complete dominance and controls normal visceral asymmetry. Absence of this control allows the situs of visceral asymmetry to be determined in a random fashion. This hypothesis also appears to apply to the inheritance of situs inversus in man and to the experimental production of situs inversus.