Dissecting the genetic basis of neurodevelopmental disorders and demyelinating neuropathies

The understanding of the pathophysiology of most rare, complex neurological disorders has been elusive, especially in the case of complex demyelinating neuropathies and neurodevelopmental disorders. In my work, I learnt to employ two main techniques that will help advance the search for better understanding of neurodevelopmental disorders: next generation sequencing and functional validation of rare genetic variants. The main aim of my research was to establish the genetic diagnosis in several patients affected by complex syndromes such as peripheral neuropathy with central nervous system involvement (Chapter 3), neurodevelopmental disorders (Chapter 4) and epilepsy (Chapter 5). The phenotypic and genotypic correlations of identified gene variants were investigated in these chapters and is a profound theme in my project. To achieve this, an integrated approach combining next generation sequencing (NGS) technology, homozygosity mapping, array genotyping, traditional Sanger sequencing and functional experiments was undertaken. Firstly, I describe the work performed in an attempt to identify the causative gene in a cohort of young children presented with an early-onset hereditary form of chronic inflammatory demyelinating polyneuropathy with a central and peripheral involvement. My key findings were that: i) neurofascin is the first gene causally responsible for an inherited disorder that resembles CIDP, ii) this is the largest clinical cohort to date of patients with NFASC mutations with 10 individuals, and iii) the functional evidence implicate the major protein isoforms, which were also shown to be the main targets for the autoantibodies in CIDP pathogenesis. Secondly, I describe the work done on various neurodevelopmental disorder (NDD) genes, with particular focus on a newly identified gene presenting with a complex neurodevelopmental phenotype comprised of developmental delay, epilepsy, and/or a demyelinating neuropathy. My key findings were that: i) NARS1, a cytoplasmic aminoacyl-tRNA synthetase enzyme can be causative for this disorder by either a de-novo heterozygous or a biallelic inheritance mode, ii) functional investigations showed reduced aminoacylation activity in the disease-associated biallelic mutations using fibroblasts and iNPCs transcriptomics, suggesting that the majority of NARS1 mutations cause a loss of function of the protein by reduced expression and disruption of dimer formation suggesting a loss-of-function mechanism, and iii) increased yeast growth in the disease-associated heterozygous mutations showing near normal protein expression are suggestive of a gain-of-function mechanism. Finally, I describe the work done on two relative new genes (PIGS and TARS1) in an attempt to expand the patient phenotypic spectrum, as well as an interesting candidate gene (SLITRK3) linked with epilepsy. I present my understanding for disease-gene discovery that will enable me and other members of the neurogenetics field to identify disease-mechanisms and address important gaps of translational research into rare neurological diseases such as those described in this thesis

Life Cycle and Development of Diptera

Diptera, or true flies, are of considerable economic importance, as these flies have a valuable role as scavengers, parasitoids and predators of other insects, pollinators, food for predators, bio-indicators of water quality, and tools for scientific research. In nine chapters, this book examines various aspects of flies of the order Diptera as well as some types of mosquitos and midges. Topics covered include taxonomy, phylogeny, life cycle, feeding habits, population control strategies, and more. A unique chapter on forensic entomology is particularly interesting. Beautifully illustrated and expertly researched, this volume will appeal to entomologists, biologists, and naturalists

Antioxidant and DPPH-Scavenging Activities of Compounds and Ethanolic Extract of the Leaf and Twigs of Caesalpinia bonduc L. Roxb.

Antioxidant effects of ethanolic extract of Caesalpinia bonduc and its isolated bioactive compounds were evaluated in vitro. The compounds included two new cassanediterpenes, 1α,7α-diacetoxy-5α,6β-dihydroxyl-cass-14(15)-epoxy-16,12-olide (1)and 12α-ethoxyl-1α,14β-diacetoxy-2α,5α-dihydroxyl cass-13(15)-en-16,12-olide(2); and others, bonducellin (3), 7,4’-dihydroxy-3,11-dehydrohomoisoflavanone (4), daucosterol (5), luteolin (6), quercetin-3-methyl ether (7) and kaempferol-3-O-α-L-rhamnopyranosyl-(1Ç2)-β-D-xylopyranoside (8). The antioxidant properties of the extract and compounds were assessed by the measurement of the total phenolic content, ascorbic acid content, total antioxidant capacity and 1-1-diphenyl-2-picryl hydrazyl (DPPH) and hydrogen peroxide radicals scavenging activities.Compounds 3, 6, 7 and ethanolic extract had DPPH scavenging activities with IC50 values of 186, 75, 17 and 102 μg/ml respectively when compared to vitamin C with 15 μg/ml. On the other hand, no significant results were obtained for hydrogen peroxide radical. In addition, compound 7 has the highest phenolic content of 0.81±0.01 mg/ml of gallic acid equivalent while compound 8 showed the highest total antioxidant capacity with 254.31±3.54 and 199.82±2.78 μg/ml gallic and ascorbic acid equivalent respectively. Compound 4 and ethanolic extract showed a high ascorbic acid content of 2.26±0.01 and 6.78±0.03 mg/ml respectively.The results obtained showed the antioxidant activity of the ethanolic extract of C. bonduc and deduced that this activity was mediated by its isolated bioactive compounds

IMPACT OF GRAZING ON SOIL ORGANIC MATTER AND PHYSICAL PROPERTIES OF A FLUVISOL IN NORTWEST SERBIA

The effects of long-term (>20 yr) grazing on the selected physical properties of a non carbonated silty-clay Fluvisols were studied in the region of the Kolubara Valley, Northwest Serbia. Two adjacent land-use types (native deciduous forest and natural pasture soils converted from forests for more than 20 years) were chosen for the study. Disturbed and undisturbed soil samples were collected from three sites at each of the two different land-use types from the depths of 0–15, 15–30 and 30–45 cm. In relation to the soil under native forest, soil organic matter content, total porosity and air-filled porosity were significantly reduced after long-term of grazing. The bulk density (0.99–1.48 g cm–3) and the saturated hydraulic conductivity (6.9.10–2–3.2.10–4 cm s–1) were significantly lower in forest compared to the adjacent pasture (ex-forest) soil (1.49–1.55 g cm–3 and 3.4.10–4–5.5.10–4 cm s–1, respectively). In addition, forest had significantly lower dry mean weight diameter (7.0–9.2 mm) and greater wet mean weight diameter (2.0–2.6 mm) for 0–45 cm depth compared with the pasture (8.8–9.4 mm and 1.8–2.3 mm, respectively). The decrease of soil organic matter content and reduction in aggregate stability under long-term grazing rendered the soil more susceptible to compaction. In conclusion, the results of this study indicate that removal of permanent vegetation in the conversion process from forest areas to pasture land may lead to loss of soil productivity and serious soil degradation. Obviously, there is a need for greater attention to developing sustainable land use practices in management of these ecosystems to prevent further degradation of pasture soils in the region

Association of agronomic and forage quality traits in red clover (Trifolium pratense L.)

Red clover (Trifolium pretense L.) is a source of highly nutritional voluminous forage for livestock feed. The objectives of this investigation were to: I) annotate associations of agronomic and forage quality traits of red clover accessions; II) group accessions in relation to analyzed traits. The plant material used included 46 cultivars and local populations, which originated from 17 countries, representing part of the red clover collection from the Institute of Field and Vegetable Crops in Novi Sad, Serbia. The field trial was sown during two growing seasons in Novi Sad. The agronomic traits (plant height-PH, internodes number per stem-IN, green mass yield-GMY, dry matter yield-DMY) were evaluated from the second cut in the nursery of the second year of life. Forage quality traits analyzed were content of neutral detergent fibers (NDF), acid detergent fiber (ADF), and crude protein (CP). In regard to very strong associations of PH, GMY and DMY, and to moderately strong associations of PH and IN, it could be anticipated that the indirect selection for higher plants with higher IN can lead to higher GMY and DMY. The breeding of red clover for NDF and ADF and elevated CP might be successful, but can reduce herbage yield (due to very weak positive and negative associations with GMY and DMY). The accessions were grouped to the six clusters which can facilitate selection and breeding for different agronomic and quality objectives

Bowdoin Orient v.125, no.1-25 (1994-1995)

https://digitalcommons.bowdoin.edu/bowdoinorient-1990s/1006/thumbnail.jp

Variation of Soil Structure in the Foot and Toe Slopes of Mt. Vukan, East-central Serbia

This paper presents the variation of soil structure along the foot and toe slopes of Mt. Vukan, East-Central Serbia. The analysis of aggregate size distribution and structure indices were conducted by means of soil units, characteristic soil horizons and elevation differences along the study area. Soils of Great Field located at different elevations were found to have significant variation in ASD and soil structure indices. Topsoil horizon of Eutric Cambisols have higher MWD after dry sieving, but at the same time it has the highest variation in MWD after wet sieving, indicating low water stability, which is opposite to the coefficient of aggregability. We share an opinion that change in MWD better depicts soils structure stability to water. The results of correlation analysis indicated that clay content is correlated more to structure indices compared with SOM content. SOM is significantly correlated with ASD and soil structure indices only in Calcomelansols, whereas the significant correlation of clay content and soil structure is more evident in Eutric Cambisols and Non-calcaric Chernozems, compared with other soil units. Soil structure variation along the lowest chain of Catena might be strong, and that it has to be analyzed from the point of view of soil unit and their corresponding soil horizons