Next-generation sequencing of iron-metabolism related genes in Portuguese patients with iron overload: novel pathogenic genetic variants

Objective: In Southern European countries up to one-third of the patients with hereditary hemochromatosis (HH) do not present the common HFE risk genotype. In order to investigate the molecular basis of these cases we have designed a gene panel for rapid and simultaneous analysis of 6 HH-related genes (HFE, TFR2, HJV, HAMP, SLC40A1 and FTL) by next-generation sequencing (NGS). Materials and Methods: Eighty-eight iron overload Portuguese patients, negative for the common HFE mutations, were analysed. A TruSeq Custom Amplicon kit (TSCA, by Illumina) was designed in order to generate 97 amplicons covering exons, intron/exon junctions and UTRs of the mentioned genes with a cumulative target sequence of 12115bp. Amplicons were sequenced in the MiSeq instrument (IIlumina) using 250bp paired-end reads. Sequences were aligned against human genome reference hg19 using alignment and variant caller algorithms in the MiSeq reporter software. Novel variants were validated by Sanger sequencing and their pathogenic significance were assessed by in silico studies. Results: We found a total of 55 different genetic variants. These include novel pathogenic missense and splicing variants (in HFE and TFR2), a very rare variant in IRE of FTL, a variant that originates a novel translation initiation codon in the HAMP gene, among others. Conclusion: The merging of TSCA methodology and NGS technology appears to be an appropriate tool for simultaneous and fast analysis of HH-related genes in a large number of samples. However, establishing the clinical relevance of NGS-detected variants for HH development remains a hard-working task, requiring further functional studies.

Description and geochemistry of DSDP Leg 55 Holes

One of the objectives of Leg 55 was to investigate the Tertiary history of sedimentation and environment on the Emperor Seamounts after their volcanic activity. For the three first sites, 430, 431, and 432, drilled on Ojin, Nintoku, and Yömei Seamounts, the Neogene sedimentary deposits are not well represented and are not typical pelagic sediments. Except for two holes (430A and 432), where we found calcareous oozes, the sediments are heterogeneous sands, gravels, and pebbly mudstones with a wide range in grain size and composition. Two phenomena characterize these deposits: the inheritance of volcaniclastic material and its alteration, and the authigenesis of secondary minerals including silicates, phosphates, and ferromanganese oxides formed under volcanic influence in a marine environment.

Evaluation of ultrafiltration to recover aqueous iron phosphating/degreasing bath /

"Contract no. CR-815829."

The Vermilion iron-bearing district of Minnesota, with an atlas;

"Résumé of literature": p. 55-128.

Iron concentration and fluxes in the water column during POLARSTERN cruises in the Southern Ocean

In large parts of the Southern Ocean, primary production is limited due to shortage of iron (Fe). We measured vertical Fe profiles in the western Weddell Sea, Weddell-Scotia Confluence, and Antarctic Circumpolar Current (ACC), showing that Fe is derived from benthic Fe diffusion and sediment resuspension in areas characterized by high turbulence due to rugged bottom topography. Our data together with literature data reveal an exponential decrease of dissolved Fe (DFe) concentrations with increasing distance from the continental shelves of the Antarctic Peninsula and the western Weddell Sea. This decrease can be observed 3500 km eastward of the Antarctic Peninsula area, downstream the ACC. We estimated DFe summer fluxes into the upper mixed layer of the Atlantic sector of the Southern Ocean and found that horizontal advection dominates DFe supply, representing 54 ± 15% of the total flux, with significant vertical advection second most important at 29 ± 13%. Horizontal and vertical diffusion are weak with 1 ± 2% and 1 ± 1%, respectively. The atmospheric contribution is insignificant close to the Antarctic continent but increases to 15 ± 10% in the remotest waters (>1500 km offshore) of the ACC. Translating Southern Ocean carbon fixation by primary producers into biogenic Fe fixation shows a twofold excess of new DFe input close to the Antarctic continent and a one-third shortage in the open ocean. Fe recycling, with an estimated 'fe' ratio of 0.59, is the likely pathway to balance new DFe supply and Fe fixation.

Susceptibility, d13C and Iron of Site 171-1051

Current models of the global carbon cycle lack natural mechanisms to explain known large, transient shifts in past records of the stable carbon-isotope ratio (delta13C) of carbon reservoirs. The injection into the atmosphere of ~1,200-2,000 gigatons of carbon, as methane from the decomposition of sedimentary methane hydrates, has been proposed to explain a delta13C anomaly associated with high-latitude warming and changes in marine and terrestrial biota near the Palaeocene-Eocene boundary, about 55 million years ago. These events may thus be considered as a natural 'experiment' on the effects of transient greenhouse warming. Here we use physical, chemical and spectral analyses of a sediment core from the western North Atlantic Ocean to show that two-thirds of the carbon-isotope anomaly occurred within no more than a few thousand years, indicating that carbon was catastrophically released into the ocean and atmosphere. Both the delta13C anomaly and biotic changes began between 54.93 and 54.98 million years ago, and are synchronous in oceans and on land. The longevity of the delta13C anomaly suggests that the residence time of carbon in the Palaeocene global carbon cycle was ~120 thousand years, which is similar to the modelled response after a massive input of methane. Our results suggest that large natural perturbations to the global carbon cycle have occurred in the past-probably by abrupt failure of sedimentary carbon reservoirs-at rates that are similar to those induced today by human activity.

Conventional and 40Ar/39Ar K-Ar ages of volcanic rocks at DSDP Leg 55 Holes

Conventional K-Ar, 40Ar/39Ar total fusion, and 40Ar/39Ar incremental heating data on hawaiite and tholeiitic basalt samples from Ojin (Site 430), alkalic basalt samples from Nintoku (Site 432), and alkalic and tholeiitic basalt samples from Suiko (Site 433) seamounts in the Emperor Seamount chain give the following best ages for these volcanoes: Ojin = 55.2 ± 0.7 m.y., Nintoku = 56.2 ± 0.6 m.y., and Suiko = 64.7 ± 1.1 m.y. These new data bring to 27 the number of dated volcanoes in the Hawaiian-Emperor volcanic chain. The new dates prove that the age progression from Kilauea Volcano on Hawaii (0 m.y.) through the Hawaiian-Emperor bend (- 43 m.y.) to Koko Seamount (48.1 m.y.) in the southernmost Emperor Seamounts continues more than halfway up the Emperor chain to Suiko Seamount. The age versus distance data for the Hawaiian-Emperor chain are consistent with the kinematic hot-spot hypothesis, which predicts that the volcanoes are progressively older west and north away from the active volcanoes of Kilauea and Mauna Loa. The data are consistent with an average volcanic propagation velocity of either 8 cm/year from Suiko to Kilauea or of 6 cm/year from Suiko to Midway followed by a velocity of 9 cm/year from Midway to Kilauea, but it appears that the change in direction that formed the Hawaiian- Emperor bend probably was not accompanied by a major change in velocity.