Genetic variants modify the effect of age on APOE methylation in the Genetics of Lipid Lowering Drugs and Diet Network study

Although apolipoprotein E (APOE) variants are associated with age-related diseases, the underlying mechanism is unknown and DNA methylation may be a potential one. With methylation data, measured by the Infinium Human Methylation 450 array, from 993 participants (age ranging from 18 to 87 years) in the Genetics of Lipid Lowering Drugs and Diet Network (GOLDN) study, and from Encyclopedia of DNA Elements (ENCODE) consortium, combined with published methylation datasets, we described the methylation pattern of 13 CpG sites within APOE locus, their correlations with gene expression across cell types, and their relationships with age, plasma lipids, and sequence variants. Based on methylation levels and the genetic regions, we categorized the 13 APOE CpG sites into three groups: Group 1 showed hypermethylation (> 50%) and were located in the promoter region, Group 2 exhibited hypomethylation (< 50%) and were located in the first two exons and introns, and Group 3 showed hypermethylation (> 50%) and were located in the exon 4. APOE methylation was negatively correlated with gene expression (minimum r = −0.66, P = 0.004). APOE methylation was significantly associated with age (minimum P = 2.06E-08) and plasma total cholesterol (minimum P = 3.53E-03). Finally, APOE methylation patterns differed across APOE ε variants (minimum P = 3.51E-05) and the promoter variant rs405509 (minimum P = 0.01), which further showed a significant interaction with age (P = 0.03). These findings suggest that methylation may be a potential mechanistic explanation for APOE functions related to aging and call for further molecular mechanistic studies.     

Description of a new genus and three new species of Otothyrinae (Siluriformes,Loricariidae)

The genus Hisonotus was resurrected as a member of the tribe Otothyrini (actually subfamily Otothyrinae). However, phylogenetic studies based on morphological and molecular data showed that Hisonotus is not monophyletic and independent lineages can be identified, such as the group composed of the species Hisonotus insperatus, Hisonotus luteofrenatus, Hisonotus oliveirai, Hisonotus paresi and Hisonotus piracanjuba, a lineage unrelated to that containing the type species of the genus Hisonotus (Hisonotus notatus). Herein, based in molecular and morphological data, a new genus is described to accommodate the lineage mentioned above, into which are also added three new species. This new genus can be distinguished from other genera of Otothyrinae by the following combination of characters: (1) a pair of rostral plates at the tip of the snout; (2) two large pre-nasal plates just posterior to the rostral plates; (3) a supra-opercular plate that receives the laterosensory canal from the compound pterotic before the preopercle; (4) a well developed membrane at anal opening in females; and (5) a V-shaped spinelet. A key to species of Curculionichthys is provided.     

Annotated type catalogue of the Megaspiridae,Orthalicidae, and Simpulopsidae (Mollusca,Gastropoda, Orthalicoidea) in the Natural History Museum,London

The type status is described for 65 taxa of the Orthalicoidea, classified within the families Megaspiridae (14), Orthalicidae (30), and Simpulopsidae (20); one taxon is considered a nomen inquirendum. Lectotypes are designated for the following taxa: Helix brephoides d’Orbigny, 1835; Simpulopsis cumingi Pfeiffer, 1861; Bulimulus (Protoglyptus) dejectus Fulton, 1907; Bulimus iris Pfeiffer, 1853. The type status of Bulimus salteri Sowerby III, 1890, and Strophocheilus (Eurytus) subirroratus da Costa, 1898 is now changed to lectotype according Art. 74.6 ICZN. The taxa Bulimus loxostomus Pfeiffer, 1853, Bulimus marmatensis Pfeiffer, 1855, Bulimus meobambensis Pfeiffer, 1855, and Orthalicus powissianus var. niveus Preston 1909 are now figured for the first time. The following taxa are now considered junior subjective synonyms: Bulimus marmatensis Pfeiffer, 1855 = Helix (Cochlogena) citrinovitrea Moricand, 1836; Vermiculatus Breure, 1978 = Bocourtia Rochebrune, 1882. New combinations are: Kuschelenia (Bocourtia) Rochebrune, 1882; Kuschelenia (Bocourtia) aequatoria (Pfeiffer, 1853); Kuschelenia (Bocourtia) anthisanensis (Pfeiffer, 1853); Kuschelenia (Bocourtia) aquila (Reeve, 1848); Kuschelenia (Bocourtia) badia (Sowerby I, 1835); Kuschelenia (Bocourtia) bicolor (Sowerby I, 1835); Kuschelenia (Bocourtia) caliginosa (Reeve, 1849); Kuschelenia (Bocourtia) coagulata (Reeve, 1849); Kuschelenia (Bocourtia) cotopaxiensis (Pfeiffer, 1853); Kuschelenia (Bocourtia) filaris (Pfeiffer, 1853); Kara indentata (da Costa, 1901); Clathrorthalicus magnificus (Pfeiffer, 1848); Simpulopsis (Eudioptus) marmartensis (Pfeiffer, 1855); Kuschelenia (Bocourtia) nucina (Reeve, 1850); Kuschelenia (Bocourtia) ochracea (Morelet, 1863); Kuschelenia (Bocourtia) peaki (Breure, 1978); Kuschelenia (Bocourtia) petiti (Pfeiffer, 1846); Clathrorthalicus phoebus (Pfeiffer, 1863); Kuschelenia (Bocourtia) polymorpha (d’Orbigny, 1835); Scholvienia porphyria (Pfeiffer, 1847); Kuschelenia (Bocourtia) purpurata (Reeve, 1849); Kuschelenia (Bocourtia) quechuarum Crawford, 1939; Quechua salteri (Sowerby III, 1890); Kuschelenia (Bocourtia) subfasciata Pfeiffer, 1853; Clathrorthalicus victor (Pfeiffer, 1854). In an addedum a lectotype is being designated for Bulimulus (Drymaeus) interruptus var. pallidus Preston, 1909. An index is included to all taxa mentioned in this paper and the preceding ones in this series (Breure and Ablett 2011, 2012, 2014).     

The complex translocation (9;14;14) involving IGH and CEBPE genes suggests a new subgroup in B-lineage acute lymphoblastic leukemia

Many subtypes of acute lymphoblastic leukemia (ALL) are associated with specific chromosomal rearrangements. The complex translocation t(9;14;14), a variant of the translocation (14;14)(q11;q32), is a rare but recurrent chromosomal abnormality involving the immunoglobulin heavy-chain (IGH) and CCAAT enhancer-binding protein (CEBPE) genes in B-lineage ALL (B-ALL) and may represent a new B-ALL subgroup. We report here the case of a 5-year-old girl with B-ALL, positive for CD19, CD38 and HLA-DR. A direct technique and G-banding were used for chromosomal analysis and fluorescentin situ hybridization (FISH) with BAC probes was used to investigate a possible rearrangement of the IGH andCEBPE genes. The karyotype exhibit the chromosomal aberration 46,XX,del(9)(p21),t(14;14)(q11;q32). FISH with dual-color break-apartIGH-specific and CEPBE-specific bacterial artificial chromosome (BAC) probes showed a complex t(9;14;14) associated with a deletion of cyclin-dependent kinase inhibitor 2A (CDKN2A) and paired box gene 5 (PAX5) at 9p21-13 and duplication of the fusion gene IGH-CEBPE.     

Putative Virulence Genes and Biofilm Production Among Typical Enteroaggregative Escherichia coli Isolates from Diarrhoeic Children in Kashmir and Andhra Pradesh

Fifty-eight typical EAEC isolates from children with diarrhoea were examined for HEp-2 cell adherence assay, presence of dispersin (aap), yersiniabactin (irp2), plasmid encoded toxins (pet), Shigella enterotoxin1 (set1A) and cryptic open reading frame (shf) putative virulence genes by polymerase chain reaction as well as for biofilm production. All the isolates showed aggregative adherence pattern on HEp-2 cells. All but five isolates (91.3 %) carried aap gene. While irp2, pet, set1A and shf genes were detected in 68.9, 5.1, 39.6, and 60.3 % isolates, respectively. Thirty-three (64.7 %) isolates out of 51 tested were found to produce biofilm which was found to be significantly associated only with set1A virulence gene (P = 0.025). Highest amount of biofilm was produced by a strain that possessed all the genes studied. Out of 14 isolates in which the most frequent gene combination (aap, irp2 and shf) was observed, only six produced biofilm. It is concluded that there is significant heterogeneity in putative virulence genes of EAEC isolates from diarrhoeic children and biofilm formation is associated with multiple genes.     

Two common and problematic leucochrysine species – Leucochrysa (Leucochrysa) varia (Schneider) and L. (L.) pretiosa (Banks) (Neuroptera,Chrysopidae): redescriptions and synonymies

We dedicate this article to the memory of Sergio de Freitas, FCAV-UNESP, Jaboticabal, São Paulo, Brazil (deceased, 2012). He was an active and enthusiastic Neuropterist and the cherished mentor and friend of Francisco Sosa.Leucochrysa McLachlan is the largest genus in the Chrysopidae, yet it has received relatively little taxonomic attention. We treat two problematic and common Leucochrysa species – Leucochrysa (Leucochrysa) varia (Schneider, 1851) and Leucochrysa (Leucochrysa) pretiosa (Banks, 1910). Both are highly variable in coloration and were described before the systematic importance of chrysopid genitalia was recognized. Recent studies show that these species occur within a large complex of cryptic species and that they have accumulated a number of taxonomic problems. We identify new synonymies for each of the species–for Leucochrysa (Leucochrysa) varia: Leucochrysa (Leucochrysa) ampla (Walker, 1853), Leucochrysa internata (Walker, 1853), and Leucochrysa (Leucochrysa) walkerina Navás, 1913; for Leucochrysa (Leucochrysa) pretiosa: Leucochrysa (Leucochrysa) erminea Banks, 1946. The synonymy of Leucochrysa delicata Navás, 1925 with Leucochrysa (Leucochrysa) pretiosa is stabilized by the designation of a neotype. The following species, which were previously synonymized with Leucochrysa (Leucochrysa) varia or Leucochrysa (Leucochrysa) pretiosa, are reinstated as valid: Leucochrysa (Leucochrysa) phaeocephala Navás, 1929, Leucochrysa (Leucochrysa) angrandi (Navás, 1911), and Leucochrysa (Leucochrysa) variata (Navás, 1913). To help stabilize Leucochrysa taxonomy, lectotypes are designated for Allochrysa pretiosa and Allochrysa variata. Finally, Leucochrysa vegana Navás, 1917 is considered a nomen dubium.     

Profundulus kreiseri,a new species of Profundulidae (Teleostei,Cyprinodontiformes) from northwestern?Honduras

A new species of Profundulus, Profundulus kreiseri (Cyprinodontiformes: Profundulidae), is described from the Chamelecón and Ulúa Rivers in the northwestern Honduran highlands. Based on a phylogenetic analysis using cytochrome b and the presence of synapomorphic characters (dark humeral spot, a scaled preorbital region and between 32-34 vertebrae), this new species is placed in the subgenus Profundulus, which also includes Profundulus (Profundulus) oaxacae, Profundulus (Profundulus) punctatus and Profundulus (Profundulus) guatemalensis. Profundulus kreiseri can be distinguished from other members of the subgenus Profundulus by having less than half of its caudal fin densely scaled. Profundulus kreiseri can further be differentiated from Profundulus (Profundulus) oaxacae and Profundulus (Profundulus) punctatus by the absence of rows of dark spots on its flanks. The new species can further be differentiated from Profundulus (Profundulus) guatemalensis by the presence of fewer caudal- and pectoral-fin rays. The new species is distinguished from congeners of the profundulid subgenus Tlaloc (viz., Profundulus (Tlaloc) hildebrandi, Profundulus (Tlaloc) labialis, Profundulus (Tlaloc) candalarius and Profundulus (Tlaloc) portillorum) by having a scaled preorbital region and a dark humeral spot. Profundulus kreiseri and Profundulus portillorum are the only two species of Profundulus that are endemic to the region south of the Motagua River drainage in southern Guatemala and northwestern Honduras.     

A Critical Component of Meiotic Drive in Neurospora Is Located Near a Chromosome Rearrangement

Neurospora fungi harbor a group of meiotic drive elements known as Spore killers (Sk). Spore killer-2 (Sk-2) and Spore killer-3 (Sk-3) are two Sk elements that map to a region of suppressed recombination. Although this recombination block is limited to crosses between Sk and Sk-sensitive (Sk^S) strains, its existence has hindered Sk characterization. Here we report the circumvention of this obstacle by combining a classical genetic screen with next-generation sequencing technology and three-point crossing assays. This approach has allowed us to identify a novel locus called rfk-1, mutation of which disrupts spore killing by Sk-2. We have mapped rfk-1 to a 45-kb region near the right border of the Sk-2 element, a location that also harbors an 11-kb insertion (Sk-2^INS1) and part of a >220-kb inversion (Sk-2^INV1). These are the first two chromosome rearrangements to be formally identified in a Neurospora Sk element, providing evidence that they are at least partially responsible for Sk-based recombination suppression. Additionally, the proximity of these chromosome rearrangements to rfk-1 (a critical component of the spore-killing mechanism) suggests that they have played a key role in the evolution of meiotic drive in Neurospora.     

A review of the threadfin breams of the genusNemipterus (Nemipteridae) from Japan and Taiwan,with description of a new species

Species of the fish genusNemipterus (Nemipteridae) from Japan and Taiwan are reviewed. A key, diagnoses and synonymies are provided for 9 species, including a new species:N. aurora sp. nov. (previously misidentified asN. delagoae Smith);N. bathybius Snyder;N. furcosus (Valenciennes) [mistakenly referred to asN. peronii (Valenciennes)];N. hexodon (Quoy et Gaimard);N. japonicus (Bloch);N. peronii (Valenciennes) [previously referred to asN. tolu (Valenciennes), a junior synonym];N. thosaporni Russell [previously misidentified asN. marginatus (Valenciennes)];N. virgatus (Houttuyn); andN. zysron (Bleeker) [previously referred to asN. metopias (Bleeker), a junior synonym].N. aurora is described from specimens from Taiwan, the Gulf of Thailand, East Malaysia and Indonesia. It appears to be closely allied toN. bipunctatus (Ehrenberg) (N. delagoae is a junior synonym), an Indian Ocean species, but the two species are readily distinguished on the basis of colour pattern of the dorsal and anal fins:N. aurora has a broad orange-yellow submedial stripe along the dorsal fin, and a lemon submedial stripe on the anal fin; whereas inN. bipunctatus the dorsal fin is uniformly rosy, and the anal fin has 2–4 wavy yellow stripes.     

Contribution to the knowledge of the Carabus (Archiplectes) satyrus Kurnakov, 1962, species complex in Abkhazia (Coleoptera,Carabidae, Carabini)

This study is based on a comparative analysis of extensive material of Carabus (Archiplectes) satyrus Kurnakov, 1962, its various forms and related taxa recently collected by the authors and some other collectors in Abkhazia. The status or specific affiliations of several subspecies are changed and a subspecies is described. Carabus (Archiplectes) besleticus Kurnakov, 1972, stat. n. is treated as a separate species housing six hitherto established subspecies in addition to the nominal type: Carabus (Archiplectes) besleticus mtsaranus Kurnakov, 1972, Carabus (Archiplectes) besleticus duripshensis Kurnakov, 1972, Carabus (Archiplectes) besleticus napraensis Belousov & Zamotajlov, 1993, Carabus (Archiplectes) besleticus dsychvensis Kurnakov, 1972, Carabus (Archiplectes) besleticus adzinbai Retezár, 2013, and Carabus (Archiplectes) besleticus resheviensis subsp. n. Carabus (Archiplectes) satyrus is treated as monotypical while the specific status of Carabus (Archiplectes) pseudopshuensis Zamotajlov, 1991, earlier proposed by Fominykh and Zamotajlov (2012), is confirmed based on the morphological and morphometric data.     

The heteromorph ammonite genus Ammonitoceras Dumas, 1876 (Ancyloceratidae) in the lower Aptian (Lower Cretaceous) of the Les Ferres Aptian Basin (southeastern France)

Ammonitoceras Dumas, 1876 is a genus of heteromorph ammonites characterized by the presence of a peculiar ontogenetic stage in its inner whorls: the Ammonitoceras stage. But in spite of its wide paleogeographic and biostratigraphic extension throughout the Aptian (Lower Cretaceous), this genus remains poorly known. In the present work we study specimens of Ammonitoceras from the lower Aptian Deshayesites multicostatus (Deshayesites deshayesi Zone) to Dufrenoyia furcata (Dufrenoyia furcata Zone) subzones of the Les Ferres Aptian Basin (southeastern France). The results are as follows: (1) representatives of Ammonitoceras from this area are regarded dimorphic with criocone macroconchs and ancylocone microconchs, (2) their ontogenetic sequence is described, (3) their intraspecific variability is significant and concerns the adult size and the duration of the ontogenetic stages, especially the Ammonitoceras stage, (4) two species are recognized: the earlier Ammonitoceras ucetiae Dumas, 1876, characterized by a brief Ammonitoceras stage on average, and the latter Ammonitoceras lahuseni (Sinzow, 1906), characterized by a longer Ammonitoceras stage on average. The sample of the Deshayesites grandis Subzone (Deshayesites deshayesi Zone) is composed of specimens too fragmentary to be identified at species level.     

The pleiohomeotic gene is required for maintaining expression of genes functioning in ventral appendage formation in Drosophila melanogaster

Polycomb group (PcG) proteins are negative regulators that maintain the expression of homeotic genes and affect cell proliferation. Pleiohomeotic (Pho) is a unique PcG member with a DNA-binding zinc finger motif and was proposed to recruit other PcG proteins to form a complex. The pho null mutants exhibited several mutant phenotypes such as the transformation of antennae to mesothoracic legs. We examined the effects of pho on the identification of ventral appendages and proximo-distal axis formation during postembryogenesis. In the antennal disc of the pho mutant, Antennapedia (Antp), which is a selector gene in determining leg identity, was ectopically expressed. The homothorax (hth), dachshund (dac) and Distal-less (Dll) genes involved in proximo-distal axis formation were also abnormally expressed in both the antennal and leg discs of the pho mutant. The engrailed (en) gene, which affects the formation of the anterior-posterior axis, was also misexpressed in the anterior compartment of antennal and leg discs. These mutant phenotypes were enhanced in the mutant background of Posterior sex combs (Psc) and pleiohomeotic-like (phol), which are another PcG genes. These results suggest that pho functions in maintaining expression of genes involved in the formation of ventral appendages and the proximo-distal axis.     

The phylogenetic relationships and generic limits of finches (Fringillidae)

Phylogenetic relationships among the true finches (Fringillidae) have been confounded by the recurrence of similar plumage patterns and use of similar feeding niches. Using a dense taxon sampling and a combination of nuclear and mitochondrial sequences we reconstructed a well resolved and strongly supported phylogenetic hypothesis for this family. We identified three well supported, subfamily level clades: the Holoarctic genus Fringilla (subfamly Fringillinae), the Neotropical Euphonia and Chlorophonia (subfamily Euphoniinae), and the more widespread subfamily Carduelinae for the remaining taxa. Although usually separated in a different family-group taxon (Drepanidinae), the Hawaiian honeycreepers are deeply nested within the Carduelinae and sister to a group of Asian Carpodacus. Other new relationships recovered by this analysis include the placement of the extinct Chaunoproctus ferreorostris as sister to some Asian Carpodacus, a clade combining greenfinches (Carduelis chloris and allies), Rhodospiza and Rhynchostruthus, and a well-supported clade with the aberrant Callacanthis and Pyrrhoplectes together with Carpodacus rubescens. Although part of the large Carduelis-Serinus complex, the poorly known Serinus estherae forms a distinct lineage without close relatives. The traditionally delimited genera Carduelis, Serinus, Carpodacus, Pinicola and Euphonia are polyphyletic or paraphyletic. Based on our results we propose a revised generic classification of finches and describe a new monotypic genus for Carpodacus rubescens.     

Two new species of the genus Anisomysis (Anisomysis) (Crustacea,Mysida, Mysidae) from coral reef waters in Thailand

Two new species of Anisomysis Hansen, 1910 (Mysida, Mysidae), Anisomysis (Anisomysis) spinaintus sp. n. and Anisomysis (Anisomysis) phuketensis sp. n., from coral-reef waters in Thailand are described. Anisomysis (Anisomysis) spinaintus, collected in the Chaolao Beach, Chanthaburi Province, is distinguished from the closely allied species Anisomysis (Anisomysis) incisa Tattersall, 1936, and Anisomysis (Anisomysis) hawaiiensis Murano, 1995, by the presence of 6–9 spines on the apical cleft of telson, which are absent in the latter two allied species. The new species can also be distinguished from Anisomysis (Anisomysis) aikawai Ii, 1964, by the presence of a deep telson cleft and a large number of spines on the lateral margin of telson. Anisomysis (Anisomysis) phuketensis sp. n., collected in Ko Lon, Phuket, is distinguished from the allied species Anisomysis (Anisomysis) robustispina Panampunnayil, 1984, by having a short telson and a pair of long spines on the apical part of the telson. Keys to the subgenera and species of Anisomysis, including the two new species, are presented.     

IKZF1 deletions associate with CRLF2 overexpression leading to a poor prognosis in B-cell precursor acute lymphoblastic leukaemia

Cytokine Receptor-Like Factor 2 (CRLF2) overexpression occurs in 5-15% of B-cell precursor acute lymphoblastic leukaemia (B-ALL). In ∼50% of these cases, the mechanisms underlying this dysregulation are unknown. IKAROS Family Zinc Finger 1 (IKZF1) is a possible candidate to play a role in this dysregulation since it binds to the CRLF2 promoter region and suppresses its expression. We hypothesised that IKZF1 loss of function, caused by deletions or its short isoforms expression, could be associated with CRLF2 overexpression in B-ALL. A total of 131 paediatric and adult patients and 7 B-ALL cell lines were analysed to investigate the presence of IKZF1 deletions and its splicing isoforms expression levels, the presence of CRLF2 rearrangements or mutations, CRLF2 expression and JAK2 mutations. Overall survival analyses were performed according to the CRLF2 and IKZF1 subgroups. Our analyses showed that 25.2% of patients exhibited CRLF2 overexpression (CRLF2-high). CRLF2-high was associated with the presence of IKZF1 deletions (IKZF1del, p = 0.001), particularly with those resulting in dominant-negative isoforms (p = 0.006). Moreover, CRLF2 expression was higher in paediatric samples with high loads of the short isoform IK4 (p = 0.011). It was also associated with the occurrence of the IKZF1 plus subgroup (p = 0.004). Furthermore, patients with CRLF2-high/IKZF1del had a poorer prognosis in the RELLA05 protocol (p = 0.067, 36.1 months, 95%CI 0.0-85.9) and adult cohort (p = 0.094, 29.7 months, 95%CI 11.8–47.5). In this study, we show that IKZF1 status is associated with CRLF2-high and dismal outcomes in B-ALL patients regardless of age.     

Resolution of the long-standing controversy over the type species of the genus Pseudotypotherium (Notoungulata,Typotheria, Mesotheriidae)

Mesotheres (Notoungulata: Typotheria) are among the most common mammals found in upper Miocene to Pliocene deposits of central Argentina, including the classic type Monte Hermoso locality, which defines the Montehermosan South American Land Mammal “Age”. Nevertheless, the correct name for the mesothere species from this site has been shrouded in uncertainty for well over a century due to questions of taxonomic priority, specimen provenance, and ontogenetic changes in dental formula. Since the mesotheres from Monte Hermoso were named, three distinct species have been formally considered as the type species of the genus: (1) Pseudotypotherium bravardi; (2) “Pseudotypotherium” maendrum; and (3) Pseudotypotherium exiguum. However, none of these species is a nominal species of the Pseudotypotherium genus; all three were originally referred to Typotherium. Article 67.2 of the International Code of Zoological Nomenclature (ICZN, 1999) indicates that only species considered as nominal species are eligible to set the type; in the case of Pseudotypotherium, these include: P. pulchrum, P. carlesi, P. hystatum, and P. carhuense. We conclude that Pseudotypotherium pulchrum F. Ameghino, 1904 (holotype MACN A 10299, Museo Argentino de Ciencias Naturales “Bernardino Rivadavia”, Ameghino Collection), is the type species of the mesotheriid notoungulate genus from Monte Hermoso. According to Article 68.2, F. Ameghino fixed the type by original designation in 1904 when he described P. pulchrum and included “n. g., n. sp.”. Two of the other species previously considered species P. (= T.) bravardi and P. (= T.) exiguum are invalid as type species according to Article 70.2, since their designations overlooked the previous type fixation. The third species (M. (= T.) maendrum) represents a different mesothere genus (Mesotherium) that only occurs in younger (Pleistocene) deposits. Our analysis puts an end to a historical debate that has been ongoing for more than a century regarding the identity of this well-represented late Miocene–Pliocene mesotheriine genus (Pseudotypotherium). This study provides a solid taxonomic foundation for future studies on intraspecific and ontogenetic variation of Pseudotypotherium pulchrum.     

Stereoselective Formation and Metabolism of 4-Hydroxy-Retinoic Acid Enantiomers by Cytochrome P450 Enzymes

All-trans-retinoic acid (atRA), the major active metabolite of vitamin A, plays a role in many biological processes, including maintenance of epithelia, immunity, and fertility and regulation of apoptosis and cell differentiation. atRA is metabolized mainly by CYP26A1, but other P450 enzymes such as CYP2C8 and CYP3As also contribute to atRA 4-hydroxylation. Although the primary metabolite of atRA, 4-OH-RA, possesses a chiral center, the stereochemical course of atRA 4-hydroxylation has not been studied previously. (4S)- and (4R)-OH-RA enantiomers were synthesized and separated by chiral column HPLC. CYP26A1 was found to form predominantly (4S)-OH-RA. This stereoselectivity was rationalized via docking of atRA in the active site of a CYP26A1 homology model. The docked structure showed a well defined niche for atRA within the active site and a specific orientation of the β-ionone ring above the plane of the heme consistent with stereoselective abstraction of the hydrogen atom from the pro-(S)-position. In contrast to CYP26A1, CYP3A4 formed the 4-OH-RA enantiomers in a 1:1 ratio and CYP3A5 preferentially formed (4R)-OH-RA. Interestingly, CYP3A7 and CYP2C8 preferentially formed (4S)-OH-RA from atRA. Both (4S)- and (4R)-OH-RA were substrates of CYP26A1 but (4S)-OH-RA was cleared 3-fold faster than (4R)-OH-RA. In addition, 4-oxo-RA was formed from (4R)-OH-RA but not from (4S)-OH-RA by CYP26A1. Overall, these findings show that (4S)-OH-RA is preferred over (4R)-OH-RA by the enzymes regulating atRA homeostasis. The stereoselectivity observed in CYP26A1 function will aid in better understanding of the active site features of the enzyme and the disposition of biologically active retinoids.