FORGE Canada Consortium: Outcomes of a 2-Year National Rare-Disease Gene-Discovery Project

Inherited monogenic disease has an enormous impact on the well-being of children and their families. Over half of the children living with one of these conditions are without a molecular diagnosis because of the rarity of the disease, the marked clinical heterogeneity, and the reality that there are thousands of rare diseases for which causative mutations have yet to be identified. It is in this context that in 2010 a Canadian consortium was formed to rapidly identify mutations causing a wide spectrum of pediatric-onset rare diseases by using whole-exome sequencing. The FORGE (Finding of Rare Disease Genes) Canada Consortium brought together clinicians and scientists from 21 genetics centers and three science and technology innovation centers from across Canada. From nation-wide requests for proposals, 264 disorders were selected for study from the 371 submitted; disease-causing variants (including in 67 genes not previously associated with human disease; 41 of these have been genetically or functionally validated, and 26 are currently under study) were identified for 146 disorders over a 2-year period. Here, we present our experience with four strategies employed for gene discovery and discuss FORGE’s impact in a number of realms, from clinical diagnostics to the broadening of the phenotypic spectrum of many diseases to the biological insight gained into both disease states and normal human development. Lastly, on the basis of this experience, we discuss the way forward for rare-disease genetic discovery both in Canada and internationally.     

Amino acid substitutions in the human genome: evolutionary implications of single nucleotide polymorphisms

Functional differences between amino acids have long been of interest in understanding protein evolution. Several indices exist for comparing residues on the basis of their physicochemical properties and frequencies of occurrence in conserved protein alignments. Here we present a residue dissimilarity index based on coding single nucleotide polymorphisms (SNPs) in the human genome. The index represents an average, organism-wide set of differences between residues and provides important insight into evolutionary restraints on residue substitutions in the human genome. Unlike previous models, it is not restricted to highly conserved protein structures, nor confounded by evolutionary differences between species. Our results confirm earlier observations regarding residue mutabilities but also suggest that in addition to the established key properties, such as size and polarity, charge conservation may be an important and currently underestimated factor in protein evolution. We also estimate that less than 51% of amino acid substitutions occurring in the human genome are evolutionarily neutral.     

Immunohistochemical properties of nerve fibres supplying accessory male genital glands in the pig. A colocalisation study

 Immunohistochemical studies have been performed to investigate the occurrence and coexistence of two catecholamine-synthesising enzymes, tyrosine hydroxylase and dopamine-β-hydroxylase, and several neuropeptides, including neuropeptide Y, vasoactive intestinal polypeptide, Leu⁵-enkephalin, somatostatin, calcitonin gene-related peptide and substance P, in nerve fibres supplying porcine accessory genital glands, the seminal vesicles, prostate (body and the disseminated part) and bulbourethral glands. Three major populations of nerve fibres supplying non-vascular elements of the glands have been distinguished (from the largest to the smallest one): (1) noradrenergic fibres, the majority of which contain Leu⁵-enkephalin, neuropeptide Y or, to a lesser extent, somatostatin, (2) non-noradrenergic, putative cholinergic fibres containing vasoactive intestinal polypeptide, neuropeptide Y and/or somatostatin and, (3) non-noradrenergic, presumably sensory fibres, containing calcitonin gene-related peptide and substance P. Whilst the coexistence patterns within nerves supplying particular glands are similar, the density of innervation varies between the organs. The innervation of the seminal vesicles and prostatic body is more developed than that of the disseminated part of the prostate and bulbourethral glands. The majority of noradrenergic fibres related to blood vessels contain neuropeptide Y only, while the non-noradrenergic nerves contain mainly vasoactive intestinal polypeptide. The possible function and origin of particular nerve fibre populations are discussed. Accepted: 16 November 1998     

Erinnerungen an Ornithologen, die ich kannte

Zusammenfassung Auf Anregung des Journal-Herausgebers hat der Autor seinen am 28. September 1997 vor der 130. DO-G-Jahresversammlung in Neubrandenburg gehaltenen Vortrag für die nachfolgende Veröffentlichung niedergeschrieben. Er berichtet aus den Biographien mehrerer, zumeist ost- und westeuropäischer (bereits verstorbener) Ornithologen Mitte des 20. Jahrhunderts; sein Bemühen gilt insbesondere der Untersuchung des Einflusses der politisch-gesellschaftlichen Verhältnisse auf deren wissenschaftliche Tätigkeit sowie der Verstrickungen einiger dieser Wissenschaftler in die Politik. Spezielles Interesse schenkt der Autor der Person Prof. Günther Niethammers, eines der erfolgreichsten Ornithologen Deutschlands, zeitweise auch DO-G-Präsidenten, dessen Vergangenheit in jüngster Zeit mancherlei kritische Fragen aufgeworfen hat. Der Verfasser hat umfangreisches Informations- und Datenmaterial gesammelt und war bemüht, auf dieser Grundlage wahrheitsgerecht die Vorgänge der damaligen Zeit zu rekonstruieren.

---

Reflections on Ornithologists whom I used to know

Summary Written version of a presentation which was held during the Annual General Meeting of the German Ornithologists Society in 1997 in Neubrandenburg, and which received much acclaim. The author relates the personal histories of some 15 outstanding ornithologists (most from Eastern and Western Europe) now deceased who lived around the middle of the twentieth century. The scientific achievements of the people involved are barely touched upon; rather the tenoraccent of the presentation is an investigation of the impact of socio-political relationships on their scientific activities and the involvement of some of the scientists in politics. The author presents the facts not as accusations or indictments, but rather as a stimulus to the younger generation of scientists to consider the issues, in particular to think What would I have done if I had lived there or at that time?

     

De novo translocation t(Yq−; 15p+) in a malformed boy

Summary De novo translocation of an important portion of the long arm of the Y chromosome to the short arm of No. 15 was observed in a mentally defective boy exhibiting several abnormal features: cleft palate, contracted fingers, synostosis of the 4th and 5th metatarsal bones, narrow iliac wings. Endocrinological studies suggest extreme hypogonadism.     

Denervation of the porcine ovaries performed during the early luteal phase influenced morphology and function of the gonad

We studied both morphology and steroidogenic activity of ovaries in gilts after bilateral surgical denervation performed on day 3 of the estrous cycle. Blood samples were collected from day 4 of the first estrous cycle to day 11 of the subsequent cycle. Denervation resulted in a dramatic reduction in the number or in the disappearance of tyrosine hydroxylase/dopamine-beta-hydroxylase- and/or neuropeptide Y-immunoreactive nerve fibres. On day 11 of the second cycle, the number of follicles (3-6 mm in diameter) was lower (p<0.001) in the denervated ovaries, while corpora lutea were not present. Neurectomy also led to a decrease in the concentrations of progesterone, androstendione and 17beta-estradiol in the follicular fluid originated from small (1-3 mm in diameter) as well as medium-sized follicles (3-6 mm in diameter). Similar to follicular fluid, concentration of androstendione in the follicular wall of medium-sized follicles decreased in experimental gilts in comparison to that of control animals. In addition, plasma concentrations of LH and steroid hormones were lower in the control than in the experimental group. Our results show that denervation of ovaries during the early luteal phase of the estrous cycle in gilts resulted in the changes in both morphology and steroidogenic activity. These results confirm the important role of the peripheral nerves in the function of ovaries.     

Condensing and fluidizing effects of ganglioside GM1 on phospholipid films

Mixed monolayers of the ganglioside G_M1 and the lipid dipalmitoylphosphatidlycholine (DPPC) at air-water and solid-air interfaces were investigated using various biophysical techniques to ascertain the location and phase behavior of the ganglioside molecules in a mixed membrane. The effects induced by G_M1 on the mean molecular area of the binary mixtures and the phase behavior of DPPC were followed for G_M1 concentrations ranging from 5 to 70 mol %. Surface pressure isotherms and fluorescence microscopy imaging of domain formation indicate that at low concentrations of G_M1 (<25 mol %), the monolayer becomes continually more condensed than DPPC upon further addition of ganglioside. At higher G_M1 concentrations (>25 mol %), the mixed monolayer becomes more expanded or fluid-like. After deposition onto a solid substrate, atomic force microscopy imaging of these lipid monolayers showed that G_M1 and DPPC pack cooperatively in the condensed phase domain to form geometrically packed complexes that are more ordered than either individual component as evidenced by a more extended total height of the complex arising from a well-packed hydrocarbon tail region. Grazing incidence x-ray diffraction on the DPPC/G_M1 binary mixture provides evidence that ordering can emerge when two otherwise fluid components are mixed together. The addition of G_M1 to DPPC gives rise to a unit cell that differs from that of a pure DPPC monolayer. To determine the region of the G_M1 molecule that interacts with the DPPC molecule and causes condensation and subsequent expansion of the monolayer, surface pressure isotherms were obtained with molecules modeling the backbone or headgroup portions of the G_M1 molecule. The observed concentration-dependent condensing and fluidizing effects are specific to the rigid, sugar headgroup portion of the G_M1 molecule.     

Detection of Borrelia burgdorferi s.l., Anaplasma phagocytophilum and Babesia spp. in Dermacentor reticulatus ticks found within the city of Białystok,Poland—first data

Pathogens carried by ticks pose a threat to both human and animal health across the world. Typically associated with rural landscapes, ticks appear to adapt well to life in urban recreational areas. Although Dermacentor reticulatus is commonly found across Europe, data on the prevalence of pathogens in this tick species, in an urban environment, are very limited. PCR was used to examine 368 D. reticulatus individuals collected in the Zwierzyniecki Forest Nature Reserve in Bia?ystok, Poland. In total, 10.3% of ticks were infected, with Babesia spp. (9.2%), Anaplasma phagocytophilum (0.8%) and Borrelia burgdorferi sensu lato (0.3%). Rickettsia spp., Bartonella spp., and Coxiella burnetii were not detected. Sequence analysis for Babesia-positive samples identified 79.4% of them as Babesia canis, 8.8% as Babesia microti, 5.9% as Babesia spp., 2.9% as Babesia venatorum, and 2.9% as Babesia vogeli. Results obtained in this study indicate that D. reticulatus ticks found within the urban premises of the study area are infected with at least three pathogens and therefore are an important factor in public health risk for tick-borne diseases.