Structure of the imprinted mouse Snrpn gene and establishment of its parental-specific methylation pattern

The mouse Snrpn gene encodes the Smn protein, which is involved in RNA splicing. The gene maps to a region in the central part of chromosome 7 that is syntenic to the Prader–Willi/Angelman syndromes (PWS-AS) region on human chromosome 15q11-q13. The mouse gene, like its human counterpart, is imprinted and paternally expressed, primarily in brain and heart. We provide here a detailed description of the structural features and differential methylation pattern of the gene. We have identified a maternally methylated region at the 5′ end (DMR1), which correlates inversely with the Snrpn paternal expression. We also describe a region at the 3′ end of the gene (DMR2) that is preferentially methylated on the paternal allele. Analysis of Snrpn mRNA levels in a methylase-deficient mouse embryo revealed that maternal methylation of DMR1 may play a role in silencing the maternal allele. Yet both regions, DMR1 and DMR2, inherit the parental-specific methylation profile from the gametes. This methylation pattern is erased in 12.5-days postcoitum (dpc) primordial germ cells and reestablished during gametogenesis. DMR1 is remethylated during oogenesis, whereas DMR2 is remethylated during spermatogenesis. Once established, these methylation patterns are transmitted to the embryo and maintained, protected from methylation changes during embryogenesis and cell differentiation. Transfections of DMR1 and DMR2 into embryonic stem cells and injection into pronuclei of fertilized eggs reveal that embryonic cells lack the capacity to establish anew the differential methylation pattern of Snrpn. That all PWS patients lack DMR1, together with the overall high resemblance of the mouse gene to the human SNRPN, offers an excellent experimental tool to study the regional control of this imprinted chromosomal domain.

kappa-Opioid receptor in humans: cDNA and genomic cloning, chromosomal assignment, functional expression, pharmacology, and expression pattern in the central nervous system.

Using the mouse delta-opioid receptor cDNA as a probe, we have isolated genomic clones encoding the human mu- and kappa-opioid receptor genes. Their organization appears similar to that of the human delta receptor gene, with exon-intron boundaries located after putative transmembrane domains 1 and 4. The kappa gene was mapped at position q11-12 in human chromosome 8. A full-length cDNA encoding the human kappa-opioid receptor has been isolated. The cloned receptor expressed in COS cells presents a typical kappa 1 pharmacological profile and is negatively coupled to adenylate cyclase. The expression of kappa-opioid receptor mRNA in human brain, as estimated by reverse transcription-polymerase chain reaction, is consistent with the involvement of kappa-opioid receptors in pain perception, neuroendocrine physiology, affective behavior, and cognition. In situ hybridization studies performed on human fetal spinal cord demonstrate the presence of the transcript specifically in lamina II of the dorsal horn. Some divergences in structural, pharmacological, and anatomical properties are noted between the cloned human and rodent receptors.

Influence of different genotypes in the pattern of selenoprotein expression in response to Brazil nut supplementation

The micronutrient selenium is essential to human physiology. As the amino acid selenocysteine, it is inserted into selenoproteins with a wide range of functions including antioxidant capacity, thyroid hormone metabolism, improvement of immune system, brain function, fertility and reproduction. Low selenium status has been associated with increased risk for chronic diseases, such as cancer, type-2 diabetes and cardiovascular disease. In this context, several studies have been conducted in order to investigate if selenium supplementation could reduce the risk of such diseases. However, genetic variations may interfere in the response of individuals to a dietary intervention and must be considered as a important source of inter-individual variation. Therefore, this study was conducted was conducted to investigate the influence of genetic variations in selenoproteins genes on the response to an intervention with Brazil nuts, the richest source of selenium known in nature. The study included 130 healthy volunteers with both genders, aged 20 to 60 years old selected in University of São Paulo. They received nuts for 8 weeks, eating one nut a day, and did a washout period for more 8 weeks. All volunteers had a blood sampling collection every 4 weeks during 4 months, in a total of 5. The following analysis were done: anthropometric measurements, lipid profile, plasma malondialdehyde, plasma and erythrocyte Se, selenoprotein P, plasma and erythrocyte GPx activity, gene expression of GPX1, SEPP1, SELS and SEP15. The volunteers were also genotyped for SNPs rs1050450, rs3811699, rs1800699, rs713041, rs3877899, rs7579, rs34713741 and rs5845. Each unit of Brazil nut provided an average of 300 µg of selenium. All 130 volunteers completed the protocol. The concentrations of total cholesterol and glucose decreased after 8 weeks of supplementation. Moreover, HDL concentrations were higher for carriers of the variant T allele for GPX4_rs713041. The frequencies of the variant genotypes were 5,4% for rs1050450, rs3811699 e rs1088668, 10% for rs3877899, 19,2% for rs713041 e rs7579, 11,5% for rs5845 and 8,5% for rs34713741. The levels of the five biomarkers increased significantly after supplementation. In addition, erythrocyte GPx activity was influenced by rs1050450, rs713041 and rs5845; erythrocyte selenium was influenced by rs5845 and plasma selenium by rs3877899. Gene expression of GPX1, SEPP1 and SEP15 were higher after supplementation. The SNP rs1050450 influenced GPX1 mRNA expression and rs7579 influenced SEPP1 mRNA expression. Therefore, it can be concluded that the supplementation with one of Brazil nut for 8 weeks was efficient to reduce total cholesterol and glucose levels and to increase the concentrations of the main biomarkers of selenium status in healthy adults. Furthermore, our results suggest that GPX4_rs713041 might interfere on HDL concentrations and GPx1 activity, GPX1_rs1050450 might interfere on GPx1 activity, SEP15_rs5845 might interfere on GPx1 activity and erythrocyte selenium and SEPP1_3877899 might interfere on plasma Se levels. Therefore, the effect of genetic variations should be considered in future nutritional interventions evaluating the response to Brazil nut supplementation.

RNA-Seq expression profile of genes related to neurodegenerative disorders affecting the human retina

Human neurodegenerative diseases, such as Parkinson’s disease (PD) and the neuromuscular disorders called dystroglycanopathies (DGPs), cause retinal impairments. We have used RNA-Seq technology to catalog all known genes linked to PD and DGPs expressed in the human retina and quantitate their mRNA levels in terms of FPKM. We have also characterized their expression profiles in the retina by determining their exonic, intronic and exon-intron junction expression levels, as well as the alternative splicing pattern of particular genes. We believe these data could pave the way toward understanding the molecular bases of sight deficiencies associated with neurodegenerative disorders.

Pollen profile from sediment core KB1, Holzminden, Northern Germany

The Nachtigall clay pit near Holzminden, northern Germany, is located in a subrosional basin filled with 43 m of interglacial, interstadial and stadial deposits adjacent to the Weser River. The succession separates the Older Middle Terrace from the Younger Middle Terrace of the Weser River. Nachtigall core KB1 (1998) mainly contains silt and clay with intercalated peat layers. The layers of fen peat and intercalated humic silt are between 36 and 22.5 m depth. According to palynological studies, the peat layers and some humic silts were deposited during interglacial and interstadial periods marked by forest vegetation, termed Nachtigall 1 and Nachtigall 2. They are subdivided by a stadial, termed Albaxen. The peat of Nachtigall 1 is interrupted twice by silt and clay strata (Allochthonous Unit I, II) which are reworked sediments of older glacial periods, possibly of late Elsterian or early Holsteinian age. The palynological sequences of Nachtigall and Göttingen/Ottostrasse show the same pattern. Moreover, the contemporaneous pollen profiles of Nachtigall and Göttingen/Ottostrasse can be compared with the Velay pollen sequence (France). The Nachtigall core section 36-26.02 m corresponds to Bouchet 2 - Bonnefond - Bouchet 3 in Velay. The profiles of Velay and Nachtigall are independently correlated to the MIS-timescale and correspond to MIS 7c, 7b, and 7a. TIMS 230Th/U-dating shows ages ranging from 227 + 9/-8 to 201 + 15/-13 ka, which are in good agreement with the inferred MIS 7 age.

Sedimentological investigations of a sediment core profile from the continental slope of the South Orkney Islands

Sediment patterns such as texture, composition, and facies from three selected areas of the Antarctic continental margin of the Weddell Sea are discussed in relation to environmental variations of the Quaternary hydrosphere and kryosphere. Advance and retreat of ice shelves as well as oscillations in sea ice coverage are reflected by particular sediment facies. The distribution of ice-rafted detritus tracks the Antarctic Coastal Current, and the Weddell Sea Bottom water contour current can be recognized by its distinctive winnowing and erosion pattern. Distribution and abundance of biogenic sediment components are mainly controlled by duration of sea ice coverage reflecting the long-term climatic evolution.

Results of a sediment profile and water analyses from Lake Hancza in northeastern Poland

We investigated the sedimentary record of Lake Hancza (northeastern Poland) using a multi-proxy approach, focusing on early to mid-Holocene climatic and environmental changes. AMS 14C dating of terrestrial macrofossils and sedimentation rate estimates from occasional varve thickness measurements were used to establish a chronology. The onset of the Holocene at c. 11600 cal. a BP is marked by the decline of Lateglacial shrub vegetation and a shift from clastic-detrital deposition to an autochthonous sedimentation dominated by biochemical calcite precipitation. Between 10000 and 9000 cal. a BP, a further environmental and climatic improvement is indicated by the spread of deciduous forests, an increase in lake organic matter and a 1.7% rise in the oxygen isotope ratios of both endogenic calcite and ostracod valves. Rising d18O values were probably caused by a combination of hydrological and climatic factors. The persistence of relatively cold and dry climate conditions in northeastern Poland during the first one and a half millennia of the Holocene could be related to a regional eastern European atmospheric circulation pattern. Prevailing anticyclonic circulation linked to a high-pressure cell above the retreating Scandinavian Ice Sheet might have blocked the influence of warm and moist Westerlies and attenuated the early Holocene climatic amelioration in the Lake Hancza region until the final decay of the ice sheet.

The new regional pattern : industries and gardens, workshops and farms /

Includes bibliographical references.

A truck and bus tire use pattern study. Final report.

National Bureau of Standards, Tire Systems Section, Washington, D.C.

The new metal worker pattern book; a complete course of instruction in the modern methods of developing and cutting the patterns for sheet metal work, giving the principles underlying practically every problem that is likely to come up in practice and explaining the selection and use of drawing tools and linear and geometrical drawing so clearly that one, who has had no previous knowledge of arithmetic or drawing, may understand these essentials and apply them in using the 253 problems in the development of patterns by the parallel line, conical or flaring, and triangulation systems.

Mode of access: Internet.

Lockwood's dictionary of terms used in the practice of mechanical engineering, embracing those current in the drawing office, pattern shop, foundry, fitting, turning, smiths' and boiler shops, etc., etc., comprising upwards of seven thousand definitions,

Mode of access: Internet.