Mutations in the gene encoding peroxisomal alpha-methylacyl-CoA racemase cause adult-onset sensory motor neuropathy

Sensory motor neuropathy is associated with various inherited disorders including Charcot-Marie-Tooth disease, X-linked adrenoleukodystrophy/adrenomyeloneuropathy and Refsum disease. In the latter two, the neuropathy is thought to result from the accumulation of specific fatty acids. We describe here three patients with elevated plasma concentrations of pristanic acid (a branched-chain fatty acid) and C27-bile-acid intermediates. Two of the patients suffered from adult-onset sensory motor neuropathy. One patient also had pigmentary retinopathy, suggesting Refsum disease, whereas the other patient had upper motor neuron signs in the legs, suggesting adrenomyeloneuropathy. The third patient was a child without neuropathy. In all three patients we discovered a deficiency of alpha-methylacyl-CoA racemase (AMACR). This enzyme is responsible for the conversion of pristanoyl-CoA and C27-bile acyl-CoAs to their (S)-stereoisomers, which are the only stereoisomers that can be degraded via peroxisomal beta-oxidation. Sequence analysis of AMACR cDNA from the patients identified two different mutations that are likely to cause disease, based on analysis in Escherichia coli. Our findings have implications for the diagnosis of adult-onset neuropathies of unknown aetiology.     

Systematic variation in gene expression patterns in human cancer cell lines

We used cDNA microarrays to explore the variation in expression of approximately 8,000 unique genes among the 60 cell lines used in the National Cancer Institute's screen for anti-cancer drugs. Classification of the cell lines based solely on the observed patterns of gene expression revealed a correspondence to the ostensible origins of the tumours from which the cell lines were derived. The consistent relationship between the gene expression patterns and the tissue of origin allowed us to recognize outliers whose previous classification appeared incorrect. Specific features of the gene expression patterns appeared to be related to physiological properties of the cell lines, such as their doubling time in culture, drug metabolism or the interferon response. Comparison of gene expression patterns in the cell lines to those observed in normal breast tissue or in breast tumour specimens revealed features of the expression patterns in the tumours that had recognizable counterparts in specific cell lines, reflecting the tumour, stromal and inflammatory components of the tumour tissue. These results provided a novel molecular characterization of this important group of human cell lines and their relationships to tumours in vivo.     

Insertion of Inhbb into the Inhba locus rescues the Inhba-null phenotype and reveals new activin functions

The activins (dimers of betaA or betaB subunits, encoded by the genes Inhba and Inhbb, respectively) are TGF-beta superfamily members that have roles in reproduction and development. Whereas mice homozygous for the Inhba-null allele demonstrate disruption of whisker, palate and tooth development, leading to neonatal lethality, homozygous Inhbb-null mice are viable, fertile and have eye defects. To determine if these phenotypes were due to spatiotemporal expression differences of the ligands or disruption of specific ligand-receptor interactions, we replaced the region of Inhba encoding the mature protein with Inhbb, creating the allele Inhbatm2Zuk (hereafter designated InhbaBK). Although the craniofacial phenotypes of the Inhba-null mutation were rescued by the InhbaBK allele, somatic, testicular, genital and hair growth were grossly affected and influenced by the dosage and bioactivity of the allele. Thus, functional compensation within the TGF-beta superfamily can occur if the replacement gene is expressed appropriately. The novel phenotypes in these mice further illustrate the usefulness of insertion strategies for defining protein function.     

Phylogenetic relationships of native and introduced Bemisia tabaci (Homoptera:Aleyrodidae) from China and India based on mtCOI DNA sequencing and host plant comparisons

Phylogenetic relationships for Bemisia tabaci were reconstructed by analysis of a ～780 bp fragment of the mitochondrial cytochrome oxidase I (mtCOI) gene with an emphasis on geographic range and distribution among eight eudicot plant families that are common hosts of B. tabaci worldwide to elucidate key phylogeographic linkages between populations extant in China (n=31) and India (n=34). Bootstrap values for the Maximum Parsimony tree were highly robust for all major nodes involving the major Asian clade, subgroups, and sister groups within, at 92%—100%. Between-clade distances for the Southeast Asia and three other major clades, e.g. from sub-Sahara Africa, North Africa-Mediterranean, and the Americas, were approximately >16% divergent. Two major Asian subgroups (I, II) were resolved, which represented populations indigenous to the region, comprising two (I a, I b) and five (II a—e) sister groups, respectively, which diverged by 11%. Two distinct populations from sunflower in Hyderabad grouped separately within the two Asian subgroups. All other populations grouped uniquely within Asian subgroup II or I. The “B” biotype was identified in 23 collections from China at 97.3%—99.5% nucleotide identity with “B” biotype reference sequences; it was not identified in collections from India. The majority of haplotypes were associated with 3—4 plant families, with one exception that for sister group IId (sesame, India), it might be monophagous. Thus, B. tabaci from the southeastern and near eastern regions of the Asian continent comprise of a large number of ancestral, richly divergent, mostly polyphagous populations. This region is therefore hypothesized to constitute an important Old World center of diversification for the B. tabaci complex, together with sub-Saharan Africa.     

Molecular characterization and expression of the gene encoding human erythroid-potentiating activity

Erythropoietin is the primary physiological regulator of erythropoiesis; however, in vitro studies have identified another class of mediators which appear to be important in stimulating erythroid progenitors. These factors have generally been referred to as burst-promoting activities (BPA), because they stimulate the growth of early erythroid progenitors referred to as burst-forming units-erythroid (BFU-E) which give rise to colonies of up to thousands of haemoglobinized cells. We recently reported purification of a burst-promoting activity from medium conditioned by the Mo T-lymphoblast cell line infected with human T-cell lymphotropic virus type II (HTLV-II). This purified glycoprotein of relative molecular mass (Mr) 28,000 also stimulates colony formation by more mature erythroid precursors (CFU-E) and is therefore referred to as erythroid-potentiating activity (EPA). Purified EPA specifically stimulates human and murine cells of the erythroid lineage, unlike murine interleukin-3 (IL-3) which stimulates precursor cells from all haematopoietic lineages. We report here the isolation of a complementary DNA molecular clone encoding EPA and its use in producing EPA in COS (monkey) cells and CHO (Chinese hamster ovary) cells. We also define the organization of the EPA gene in human DNA.     

Inter-simple sequence repeat (ISSR) variation in three populations of Gaura neomexicana ssp. coloradensis (Onagraceae), F.E. Warren Air Force Base, Cheyenne, Wyoming

We investigated genetic variation within and between 3 populations of Gaura neomexicana ssp. coloradensis , a federally listed threatened species, using inter-simple sequence repeats. The data matrix included 24 individuals scored for 88 bands, with 3.4% missing data. Cluster analysis showed that members of the 3 populations are very similar and are intermixed in the phenogram. Principal coordinate analysis indicated that members of the Crow Creek population are different from the Diamond Creek and ""unnamed drainage"" populations. This could be the result of historic differences, current changes in vegetation at the Crow Creek site, or an artifact of limited sampling. In any case, for management purposes it is best to preserve the dwindling population at Crow Creek as well as the thriving populations at the other 2 sites.