Retinoic acid receptor beta and angiopoietin-like protein 1 are involved in the regulation of human androgen biosynthesis

Androgens are essential for sexual development and reproduction. However, androgen regulation in health and disease is poorly understood. We showed that human adrenocortical H295R cells grown under starvation conditions acquire a hyperandrogenic steroid profile with changes in steroid metabolizing enzymes HSD3B2 and CYP17A1 essential for androgen production. Here we studied the regulatory mechanisms underlying androgen production in starved H295R cells. Microarray expression profiling of normal versus starved H295R cells revealed fourteen differentially expressed genes; HSD3B2, HSD3B1, CYP21A2, RARB, ASS1, CFI, ASCL1 and ENC1 play a role in steroid and energy metabolism and ANGPTL1, PLK2, DUSP6, DUSP10 and FREM2 are involved in signal transduction. We discovered two new gene networks around RARB and ANGPTL1, and show how they regulate androgen biosynthesis. Transcription factor RARB stimulated the promoters of genes involved in androgen production (StAR, CYP17A1 and HSD3B2) and enhanced androstenedione production. For HSD3B2 regulation RARB worked in cooperation with Nur77. Secretory protein ANGPTL1 modulated CYP17A1 and DUSP6 expression by inducing ERK1/2 phosphorylation. By contrast, our studies revealed no evidence for hormones or cell cycle involvement in regulating androgen biosynthesis. In summary, these studies establish a firm role for RARB and ANGPTL1 in the regulation of androgen production in H295R cells.

LRH-1 May Rescue SF-1 Deficiency for Steroidogenesis: An in vitro and in vivo Study

Steroidogenic factor 1 (NR5A1/SF-1) mutations usually manifest in 46,XY individuals with variable degrees of disordered sex development and in 46,XX women with ovarian insufficiency. So far, there is no genotype-phenotype correlation. The broad spectrum of phenotype with NR5A1 mutations may be due to a second hit in a gene with similar function to NR5A1/SF-1. Liver receptor homologue-1 (LRH-1/NR5A2) might be a good candidate. We performed in vitro studies for the interplay between SF-1, LRH-1 and DAX-1, expression profiles in human steroidogenic tissues, and NR5A2 genetic studies in a cohort (11 patients, 8 relatives, 11 families) harboring heterozygote NR5A1/SF-1 mutations. LRH-1 isoforms transactivate the CYP17A1 and HSD3B2 promoters similarly to SF-1 and compensate for SF-1 deficiency. DAX-1 inhibits SF-1- and LRH-1-mediated transactivation. LRH-1 is found expressed in human adult and fetal adrenals and testes. However, no NR5A2/LRH-1 mutations were detected in 14 individuals with heterozygote NR5A1/SF-1 mutations. These findings demonstrate that in vitro LRH-1 can act like SF-1 and compensate for its deficiency. Expression of LRH-1 in fetal testis suggests a role in male gonadal development. However, as we found no NR5A2/LRH-1 mutations, the 'second genetic hit' in SF-1 patients explaining the broad phenotypic variability remains elusive.

Natural history of growth hormone deficiency in a pediatric cohort.

BACKGROUND/AIMS Controversies still exist regarding the evaluation of growth hormone deficiency (GHD) in childhood at the end of growth. The aim of this study was to describe the natural history of GHD in a pediatric cohort. METHODS This is a retrospective study of a cohort of pediatric patients with GHD. Cases of acquired GHD were excluded. Univariate logistic regression was used to identify predictors of GHD persisting into adulthood. RESULTS Among 63 identified patients, 47 (75%) had partial GHD at diagnosis, while 16 (25%) had complete GHD, including 5 with multiple pituitary hormone deficiencies. At final height, 50 patients underwent repeat stimulation testing; 28 (56%) recovered and 22 (44%) remained growth hormone (GH) deficient. Predictors of persisting GHD were: complete GHD at diagnosis (OR 10.1, 95% CI 2.4-42.1), pituitary stalk defect or ectopic pituitary gland on magnetic resonance imaging (OR 6.5, 95% CI 1.1-37.1), greater height gain during GH treatment (OR 1.8, 95% CI 1.0-3.3), and IGF-1 level <-2 standard deviation scores (SDS) following treatment cessation (OR 19.3, 95% CI 3.6-103.1). In the multivariate analysis, only IGF-1 level <-2 SDS (OR 13.3, 95% CI 2.3-77.3) and complete GHD (OR 6.3, 95% CI 1.2-32.8) were associated with the outcome. CONCLUSION At final height, 56% of adolescents with GHD had recovered. Complete GHD at diagnosis, low IGF-1 levels following retesting, and pituitary malformation were strong predictors of persistence of GHD.

A 2-year multicentre, open-label, randomized, controlled study of growth hormone (Genotropin(®) ) treatment in very young children born small for gestational age: Early Growth and Neurodevelopment (EGN) Study

OBJECTIVE In Europe, growth hormone (GH) treatment for children born small for gestational age (SGA) can only be initiated after 4 years of age. However, younger age at treatment initiation is a predictor of favourable response. To assess the effect of GH treatment on early growth and cognitive functioning in very young (<30 months), short-stature children born SGA. DESIGN A 2-year, randomized controlled, multicentre study (NCT00627523; EGN study), in which patients received either GH treatment or no treatment for 24 months. PATIENTS Children aged 19-29 months diagnosed as SGA at birth, and for whom sufficient early growth data were available, were eligible. Patients were randomized (1:1) to GH treatment (Genotropin(®) , Pfizer Inc.) at a dose of 0·035 mg/kg/day by subcutaneous injection, or no treatment. MEASUREMENTS The primary objective was to assess the change from baseline in height standard deviation score (SDS) after 24 months of GH treatment. RESULTS Change from baseline in height SDS was significantly greater in the GH treatment vs control group at both month 12 (1·03 vs 0·14) and month 24 (1·63 vs 0·43; both P < 0·001). Growth velocity SDS was significantly higher in the GH treatment vs control group at 12 months (P < 0·001), but not at 24 months. There was no significant difference in mental or psychomotor development indices between the two groups. CONCLUSIONS GH treatment for 24 months in very young short-stature children born SGA resulted in a significant increase in height SDS compared with no treatment.

Quaternary and Paleogene calcareous nannofossils of ODP Leg 115

The occurrences of ten datum events for the Quaternary and top Pliocene nannofossils are identified at nine Leg 115 sites. A quantitative investigation of Paleogene nannofossils in 470 samples selected from 11 holes at 9 sites yielded 197 taxa, including one new species and 10 unidentified taxa that are likely to be new species. Regional differences in the timing of some biostratigraphically important events are recognized, and a set of datum events useful for biostratigra- phy in the tropical Indian Ocean is presented. Biogeographical differences are minor for Paleogene cores from the tropical sites (Sites 707-716); however, the Quaternary and late early Oligocene floras observed at the two subtropical sites (Sites 705 and 706) differ significantly from the corresponding floras of the tropical sites. Bathymetrically controlled dissolution is recognized by the reduction of species diversity in the Paleogene flora. Selective dissolution of nannofossils is also evidenced by the percentage reduction of three holococcolith taxa, Lanternithus minutus, Zygrhablithus bijugatus, and Holococcolith type A as well as by the increase of Coccolithus pelagicusand Cribrocentrum reticulatumin the deeper sites.

Late Cretaceous to Eocene calcareous nannofossil distribution near Gubbio, Italy

Using a modified sample preparation technique, we have been able to establish a detailed lower Campanian to upper Eocene nannofossil stratigraphy in the Bottaccione and Contessa Highway sections near Gubbio. Appearance and extinction levels of virtually all the commonly used calcareous nannofossil zonal markers have been recognized and can now be closely correlated with the planktonic foraminifera zonation and the magnetic reversal stratigraphy previously established in these sections. Comparisons with the nannofossil calibrations of the oceanic magnetic anomaly sequence in Deep Sea Drilling Project (DSDP) sites suggest that magmetic Subchrons C17N and C25N are missing in the Bottaccione section. The observed variability of the relative stratigraphic position of most plankton events is confirmed to less than one magnetic subchron. Absolute abundance, paleobiogeographic restriction, and differential preservation render some of the traditionally used biostratigraphic events less reliable than others.

Neogene calcareous nannofossil abundance and datums of ODP Site 186-1150 and 186-1151 sediments

Calcareous nannofossil assemblages were studied from Ocean Drilling Program Holes 1150A, 1150B, 1151A, 1151C, and 1151D in order to estimate the age of sediments drilled in the Japan Trench of the western Pacific Ocean. The abundance and species diversity of nannofossil flora are generally low but are sufficient to show that the sedimentary sequences range from Quaternary to Miocene in age (nannofossil Zones CN15-CN3). The abundance of Coccolithus pelagicus, a cold-water indicator, was studied from sediments younger than 3.83 Ma from both Holes 1150A and 1151A in order to elucidate past climate conditions. Between 3.83 and 2.82 Ma, the abundance of C. pelagicus was generally low, but abundance increased significantly after 2.82 Ma. In agreement with previous studies, this increase appears to be related to a change in the current system around the western Pacific Ocean and eastern Atlantic Ocean that occurred in response to the final elevation of the Isthmus of Panama.

Abundance of calcareous nannofossils in ODP Hole 122-761C (Table 1)

A biostratigraphically complete (all nannofossil biozones present) Cretaceous/Tertiary boundary was recovered at Site 761 on the Wombat Plateau, northwest Australian margin, during Ocean Drilling Program Leg 122. A quantitative study of calcareous nannofossils on closely spaced samples across the boundary reveals a rapid change in assemblages in a similar fashion to other Cretaceous/Tertiary boundary sites. Nannofossil species were placed into three categories: Tertiary, Cretaceous, and 'survivors'. The rapid sequential turnover in these assemblages is as follows: Cretaceous species are abruptly replaced at the boundary by opportunistic survivor species, which in turn are abruptly replaced by newly evolved Tertiary taxa. The uppermost Maestrichtian assemblage is distinctly mid-latitudinal with few Micula murus and rare to few Nephrolithus frequens. The nannofossil assemblage immediately above the boundary is dominated by an abundance bloom of Cyclagelosphaera spp. No Thoracosphaera or Braarudosphaera abundance blooms are present as at many other localities. The change from a survivor- to Tertiary-dominated assemblage is coincident with the CPla/CPlb nannofossil subzonal boundary, which is marked by the simultaneous first occurrence of several species including Cruciplacolithus tenuis and C. primus. The latter is found to first occur below C. tenuis in the most complete Cretaceous/Tertiary sections. A hiatus between Subzones CPla and CPlb is interpreted to explain this discrepancy.

Calcareous nannofossil species richness across the Paleocene-Eocene Thermal Maximum

The Paleocene-Eocene Thermal Maximum (PETM, ~5 million years ago) was an interval of global warming and ocean acidification attributed to rapid release and oxidation of buried carbon. We show that the onset of the PETM coincided with a prominent increase in the origination and extinction of calcareous phytoplankton. Yet major perturbation of the surface-water saturation state across the PETM was not detrimental to the survival of most calcareous nannoplankton taxa and did not impart a calcification or ecological bias to the pattern of evolutionary turnover. Instead, the rate of environmental change appears to have driven turnover, preferentially affecting rare taxa living close to their viable limits.

Diatom events and Neogene nannofossils of ODP Hole 201-1225A

Shipboard investigation of magnetostratigraphy and shore-based investigation of diatoms and calcareous nannofossils were used to identify datum events in sedimentary successions collected at Ocean Drilling Program (ODP) Leg 201 Site 1225. The goal was to extend the magnetic record previously studied at the same site, ODP Leg 138 Site 851, and provide a comprehensive age model for Site 1225. Two high-magnetic intensity zones at 0-70 and 200-255 meters below seafloor (mbsf) were correlated with lithologic Subunits IA and IC in Hole 1225A. Subunit IA (0-70 mbsf) contains the magnetic reversal record until the Cochiti Subchronozone (3.8 Ma) and has a sedimentation rate of 1.7 cm/k.y. This agrees with previous work done at Site 851. Subunit IC (200-255 mbsf) was not sampled at Site 851. Diatom and nannofossil biostratigraphy constrained this subunit, and we found it to contain the magnetic reversal record between Subchrons C4n.2r and C5n.2n (8.6-9.7 Ma), yielding a sedimentation rate of 2.7 cm/k.y. Biostratigraphy was used to establish the sedimentation rates within Subunits IB and ID (70-200 mbsf and 255-300 mbsf, respectively). These subunits had higher sedimentation rates (~3.4 cm/k.y.) and coincide with the late Miocene-early Pliocene biogenic bloom event (4.5-7 Ma) and the Miocene global cooling trend (10-15 Ma). High biogenic productivity associated with these subunits resulted in the pyritization of the magnetic signal. In lithologic Subunit ID, basement flow is another factor that may be altering the magnetic signal; however, the good correlation between the biostratigraphy and magnetostratigraphy indicates that the magnetic record was locked-in near the seafloor and suggests the age model is robust.

Cretaceous and Paleogene calcareous nannofossils from ODP Leg 123

Two of five holes drilled at two separate sites during Leg 123 of the Ocean Drilling Program intersected thick and relatively complete sections of Upper Cretaceous-Paleogene nannofossiliferous sediments. Although dominated by turbidite deposition in the upper part, Hole 765C contains a thick and relatively complete Albian-Oligocene section, including a particularly thick Aptian interval, with abundant and fairly well-preserved nannofossils. Several unconformities are confidently interpreted in this section that span much of the Santonian, late Campanian, Maestrichtian, late Eocene, and early Oligocene. Hole 766A contains a thick and relatively complete Albian-lower Eocene section having generally abundant and well-preserved nannofossils. Several unconformities also have been identified in this section that span much of the Coniacian, early Campanian, Maestrichtian, and late Eocene through early Pliocene. The chronostratigraphic position and length of all these unconformities may have considerable significance for reconstructing the sedimentary history and for interpreting the paleoceanography of this region. A particularly thick section of upper Paleocene-lower Eocene sediments, including a complete record across the Paleocene/Eocene boundary, also was cored in Hole 766A that contains abundant and diverse nannofossil assemblages. Although assemblages from this section were correlated successfully using a standard low-latitude zonation, difficulties were encountered that reduced biostratigraphic resolution. Several lines of evidence suggest a mid-latitude position for Site 766 during this time, including (1) high assemblage diversity characteristic of mid-latitude zones of upwelling and (2) absence of certain ecologically controlled markers found only in low latitudes.

Calcareous nannofossil stratigraphy, nannofossil events and carbon isotopic ratios of ODP Leg 183 sites

Cores from Sites 1135, 1136, and 1138 of Ocean Drilling Program Leg 183 to the Kerguelen Plateau (KP) provide the most complete Paleocene and Eocene sections yet recovered from the southern Indian Ocean. These nannofossil-foraminifer oozes and chalks provide an opportunity to study southern high-latitude biostratigraphic and paleoceanographic events, which is the primary subject of this paper. In addition, a stable isotope profile was established across the Cretaceous/Tertiary (K/T) boundary at Site 1138. An apparently complete K/T boundary was recovered at Site 1138 in terms of assemblage succession, isotopic signature, and reworking of older (Cretaceous) nannofossil taxa. There is a significant color change, a negative carbon isotope shift, and nannofossil turnover. The placement of the boundary based on these criteria, however, is not in agreement with the available shipboard paleomagnetic stratigraphy. We await shore-based paleomagnetic study to confirm or deny those preliminary results. The Paleocene nannofossil assemblage is, in general, characteristic of the high latitudes with abundant Chiasmolithus, Prinsius, and Toweius. Placed in context with other Southern Ocean sites, the biogeography of Hornibrookina indicates the presence of some type of water mass boundary over the KP during the earliest Paleocene. This boundary disappeared by the late Paleocene, however, when there was an influx of warm-water discoasters, sphenoliths, and fasciculiths. This not only indicates that during much of the late Paleocene water temperatures were relatively equable, but preliminary floral and stable isotope analyses also indicate that a relatively complete record of the late Paleocene Thermal Maximum event was recovered at Site 1135. It was only at the beginning of the middle Eocene that water temperatures began to decline and the nannofossil assemblage became dominated by cool-water species while discoaster and sphenolith abundances and diversity were dramatically reduced. One new taxonomic combination is proposed, Heliolithus robustus Arney, Ladner, and Wise.

Paleocene to middle Eocene calcareous nannofossils of Maude Rise, Weddell Sea

Cores from Sites 689 and 690 of Ocean Drilling Program Leg 113 provide the most continuous Paleocene and Eocene sequence yet recovered by deep sea drilling in the high latitudes of the Southern Ocean. The nannofossil-foraminifer oozes and chalks recovered from Maud Rise at 65°S in the Weddell Sea provide a unique opportunity for biostratigraphic study of extremely high southern latitude carbonate sediments. The presence of warm water index fossils such as the discoasters and species of the Tribrachiatus plexus facilitate the application of commonly used low latitude calcareous nannofossil biostratigraphic zonation schemes for the upper Paleocene and lower Eocene intervals. In the more complete section at Site 690, Okada and Bukry Zones CP1 through CP10 can be identified for the most part with the possible exception of Zone CP3. Several hiatuses are present in the sequence at Site 689 with the most notable being at the Cretaceous/Tertiary and Paleocene/Eocene boundaries. Though not extremely diverse, the assemblage of discoasters in the upper Paleocene and lower Eocene calcareous oozes is indicative of warm, relatively equable climates during that interval. A peak in discoaster diversity in uppermost Paleocene sediments (Zone CP8) corresponds to a negative shift in 5180 values. Associated coccolith assemblages are quite characteristic of high latitudes with abundant Chiasmolithus, Prinsius, and Toweius. Climatic cooling is indicated for middle Eocene sediments by assemblages that contain very abundant Reticulofenestra, lack common discoasters and sphenoliths and are much less diverse overall. Two new taxa are described, Biscutum? neocoronum n. sp. and Amithalithina sigmundii n. gen., n. sp.

Calcareous nannofossil stratigraphy and datums of sediments from ODP Leg 198 sites

During Ocean Drilling Program Leg 198, Sites 1207, 1208, 1212, 1213, and 1214 were drilled on Shatsky Rise, coring Lower to mid-Cretaceous successions of nannofossil chalk, porcellanite, and chert. Although recovery was poor, these sites yielded an outstanding record of calcareous nannoplankton, providing valuable data concerning the evolutionary succession and paleobiogeography of the largest Cretaceous marine habitat. Mid-Cretaceous sections (Aptian-Cenomanian) were recovered at all sites, and Site 1213 includes an apparently complete Berriasian-Hauterivian section. Biostratigraphic dating is problematic in places because of the absence or rarity of zonal fossils of both Boreal and Tethyan affinity. The majority of nannofossil assemblages are relatively typical of this age, but there are clear differences that set them apart from coeval epicontinental assemblages: for example, Lithraphidites carniolensis is common to abundant throughout and was most likely an oceanic-adapted taxon; the cold- to temperate-water species Crucibiscutum salebrosum, Repagulum parvidentatum, and Seribiscutum primitivum are entirely absent, indicating the persistence of tropical, warm surface water temperatures; and the warm-water species Hayesites irregularis is common. Most striking, however, is the virtual absence of Nannoconus and Micrantholithus, both taxa that were conspicuous and often common components of many Tethyan and Atlantic nannofloras. These forms were almost certainly neritic adapted and usually absent in deep open-ocean settings away from guyots and platforms. Other Tethyan taxa are also absent or rare and sporadically distributed (e.g., Calcicalathina oblongata, Conusphaera spp., Tubodiscus verenae, and Lithraphidites bollii), and factors related to neritic environments presumably controlled their distribution. Site 1213 also records extended Early Cretaceous ranges for species previously thought to have become extinct during the Late Jurassic (e.g., Axopodorhabdus cylindratus, Hexapodorhabdus cuvillieri, and Biscutum dorsetensis), suggesting these species became Pacific-restricted prior to their extinction. Watznaueria britannica may also have been a species with Pacific affinities before reexpansion of its biogeography in the early Aptian. One new genus (Mattiolia) and thirteen new species (Zeugrhabdotus clarus, Zeugrhabdotus petrizzoae, Helicolithus leckiei, Rhagodiscus amplus, Rhagodiscus robustus, Rhagodiscus sageri, Rhagodiscus adinfinitus, Tubodiscus bellii, Tubodiscus frankiae, Gartnerago ponticula, Haqius peltatus, Mattiolia furva, and Kokia stellata) are described from the Shatsky Rise Lower Cretaceous section.

Distribution of calcareous nannofossils in upper Cretaceous and Cenozoic sediments of the Kerguelen Plateau

ODP Leg 119 drilled 11 sites on the Kerguelen Plateau (southern Indian Ocean) and Prydz Bay (East Antarctica). Upper Pliocene through Quaternary sediments were recovered at Site 736 on the northern Kerguelen Plateau; calcareous nannofossils occurred in only a few samples. Over 700 m of middle Eocene through Quaternary sediments was cored at Site 737 on the northern Kerguelen Plateau, and calcareous nannofossils are abundant in the middle Eocene through the middle Miocene sediments. Nearly 500 m of sediments ranging from the lower Turanian to the Quaternary was recovered at Site 738 on the southern Kerguelen Plateau; calcareous nannofossils are abundant from the Miocene downward. Calcareous nannofossils are also abundant in the upper Eocene through Miocene section from Site 744 on the southern Kerguelen Plateau. Except for Core 119-746A-13H, the Neogene sequences drilled at deep-water Sites 745 and 746 off the southern Kerguelen Plateau are devoid of calcareous nannofossils. Occurrences of calcareous nannofossils were generally rare and sporadic at Sites 739 and 742 in Prydz Bay and suggest that the diamictite sequences recovered is as old as middle Eocene-early Oligocene age. Other sites drilled in Prydz Bay (Sites 740, 741, and 743) did not yield calcareous nannofossils. Species diversity of calcareous nannofossils was low (about a dozen) in the southern Indian Ocean in the Late Cretaceous. High-latitude nanno floral characteristics are apparent after the Cretaceous/Tertiary boundary extinctions. Cold climatic conditions limited Oligocene calcareous nannofossil assemblages to fewer than a dozen species, and extinctions of species generally were not compensated by originations of new species. Only a few species of calcareous nannofossils were found in the Miocene sequences, in which Coccolithuspelagicus and one or two species of Reticulofenestra exhibit extreme (0%-100%) fluctuations in assemblage dominance, and these fluctuations may reflect rapid fluctuations in the surface-water temperatures. Further deterioration of climate in the late Neogene essentially excluded calcareous nannoplankton from the Southern Ocean. Significantly warmer water conditions during part of the early-middle Pleistocene were inferred by a few lower-middle Pleistocene calcareous nannofossil species found on the Kerguelen Plateau. The calcareous nannofossil zonation of Roth (1978 doi:10.2973/dsdp.proc.44.134.1978) can be applied to the Upper Cretaceous section recovered at Site 738, and the zonation of Okada and Bukry (1980 doi:10.1016/0377-8398(80)90016-X) can be applied without much difficulty to the Paleocene to middle Eocene sequences from the Kerguelen Plateau. However, some conventional upper Paleogene markers are not useful for southern high latitudes, whereas a few nonconventional species events are useful for subdividing the upper Paleogene sequences. The latter species events include the first occurrence (FO) of Reticulofenestra reticulata, the FO and last occurrence (LO) of Reticulofenestra oamaruensis, the LO of Isthmolithus recurvus, and the LO of Chiasmolithus altus. As the Neogene sequences from the southern Indian Ocean contain only a few long-ranging, cold-water species, or are devoid of coccoliths, calcareous nannofossil zonations remain virtually unworkable for the Neogene in the high-latitude southern Indian Ocean as in other sectors of the Southern Ocean.