Structure and dynamics of the membrane attaching nitric oxide transporter nitrophorin 7

Nitrophorins represent a unique class of heme proteins that are able to perform the delicate transportation and release of the free-radical gaseous messenger nitric oxide (NO) in a pH-triggered manner. Besides its ability to bind to phospholipid membranes, the N-terminus contains an additional Leu-Pro-Gly stretch, which is a unique sequence trait, and the heme cavity is significantly altered with respect to other nitrophorins. These distinctive features encouraged us to solve the X-ray crystallographic structures of NP7 at low and high pH and bound with different heme ligands (nitric oxide, histamine, imidazole). The overall fold of the lipocalin motif is well preserved in the different X-ray structures and resembles the fold of other nitrophorins. However, a chain-like arrangement in the crystal lattice due to a number of head-to-tail electrostatic stabilizing interactions is found in NP7. Furthermore, the X-ray structures also reveal ligand-dependent changes in the orientation of the heme, as well as in specific interactions between the A-B and G-H loops, which are considered to be relevant for the biological function of nitrophorins. Fast and ultrafast laser triggered ligand rebinding experiments demonstrate the pH-dependent ligand migration within the cavities and the exit route. Finally, the topological distribution of pockets located around the heme as well as from inner cavities present at the rear of the protein provides a distinctive feature in NP7, so that while a loop gated exit mechanism to the solvent has been proposed for most nitrophorins, a more complex mechanism that involves several interconnected gas hosting cavities is proposed for NP7.

Effects of the sewage pollution in the structure and dynamics of the community of Cystoseira mediterranea (Fucales, Phaeophyceae)

We performed a study on the specific composition, structure, and dynamics of two Cystoseira mediterranea communities from the north-western Mediterranean submitted to different degrees of pollution. The structural complexity (species richness, specific distribution, and species and pattern diversity) and biomass production were lower in the polluted site. In this station, opportunistic algae (mainly Ulva rigida) loomed, and Mesophyllum lichenoides and some encrusting brown algae increased their cover. Other species (Jania rubens, and some Ceramiales) decreased their abundance when compared with the polluted site

POPULATION STRUCTURE AND PRODUCTION OF COPAIBA OLEORESIN BETWEEN VALLEYS AND HILLSIDES OF THE MINING AREA OFTROMBETAS RIVER - PARÁ1

ABSTRACT We aimed in this work to study natural populations of copaiba (Copaifera multijuga Hayne) on the Monte Branco mountain at Porto Trombetas-PA, in order to support sustainable management and the exploitation of oleoresin from copaiba. We studied the population structure of copaiba on hillsides and valleys of the south face of Monte Branco, within Saracá Taquera National Forest, where bauxite ore was extracted in the biennium 2013-2014 by Mineração Rio do Norte (MRN). We produced a 100% forest inventory of the specie and of oleoresin extraction in order to quantify the potential production of the remaining area. The density of copaiba individuals with DBH > 30 cm was 0.33 individuals per hectare in the hillside and 0.25 individuals per hectare in the valley. Both environments presented a density of 0.28 individuals per hectare. The average copaiba oleoresin yield was 0.661±0.334 liters in the hillside and 0.765±0.280 liters in the valley. The average value of both environments together (hillside and valley) was 0.714±0.218 liters. From all individuals with DBH over 30 cm, 38 (58%) produced some amount of oleoresin, averaging 1.113±0.562 liters in the hillside, 1.329±0.448 liters in the valley and 1.190±0.355 liters in both environments together. The results show the need for planning the use of the surroundings of the study area in order to reach the required volume of copaiba to make feasible the sustainable management of oleoresin extraction in the region.

Structure and function of the selectin ligand PSGL-1

P-selectin glycoprotein ligand-1 (PSGL-1) is a dimeric mucin-like 120-kDa glycoprotein on leukocyte surfaces that binds to P- and L-selectin and promotes cell adhesion in the inflammatory response. The extreme amino terminal extracellular domain of PSGL-1 is critical for these interactions, based on site-directed mutagenesis, blocking monoclonal antibodies, and biochemical analyses. The current hypothesis is that for high affinity interactions with P-selectin, PSGL-1 must contain O-glycans with a core-2 branched motif containing the sialyl Lewis x antigen (NeuAca2®3Galß1®4[Fuca1®3]GlcNAcß1®R). In addition, high affinity interactions require the co-expression of tyrosine sulfate on tyrosine residues near the critical O-glycan structure. This review addresses the biochemical evidence for this hypothesis and the evidence that PSGL-1 is an important in vivo ligand for cell adhesion.

Structure and function of the cystic fibrosis transmembrane conductance regulator

Cystic fibrosis (CF) is a lethal autosomal recessive genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR). Mutations in the CFTR gene may result in a defective processing of its protein and alter the function and regulation of this channel. Mutations are associated with different symptoms, including pancreatic insufficiency, bile duct obstruction, infertility in males, high sweat Cl-, intestinal obstruction, nasal polyp formation, chronic sinusitis, mucus dehydration, and chronic Pseudomonas aeruginosa and Staphylococcus aureus lung infection, responsible for 90% of the mortality of CF patients. The gene responsible for the cellular defect in CF was cloned in 1989 and its protein product CFTR is activated by an increase of intracellular cAMP. The CFTR contains two membrane domains, each with six transmembrane domain segments, two nucleotide-binding domains (NBDs), and a cytoplasmic domain. In this review we discuss the studies that have correlated the role of each CFTR domain in the protein function as a chloride channel and as a regulator of the outwardly rectifying Cl- channels (ORCCs).

The house that the Court built. The retained powers case law of the European Court of Justice and the constitutional structure of the Union

Euroopan unionin perustamissopimusten katsotaan muodostavan EU:n valtiosäännön, jonka rajat ovat kuitenkin perustamissopimusten määräysten väljän muotoilun sekä Euroopan unionin tuomioistuimen tulkintakäytännön valossa epätarkat. Etenkin kysymys unionin ja sen jäsenvaltioiden välisestä toimivallanjaosta on EU-oikeudellisen tutkimuksen klassikoita. Tarkastelen pro gradu -tutkielmassani unionin valtiosääntörakennetta ja unionioikeuden kokonaisvaikutusta jäsenvaltioiden toimivaltojen käyttöön erityisesti EU-tuomioistuimen ratkaisukäytännössään kehittämän niin kutsutun retained powers -doktriinin valossa. Kyseisen opin mukaan EU-oikeus asettaa vaatimuksia jäsenvaltioiden toimivaltojen käytölle myös niillä aloilla, joilla sääntelytoimivalta on jäänyt jäsenvaltioille eikä sitä ole jaettu unionin kanssa. Aiheen teoreettisen tarkastelun pohjalta analysoin Euroopaun unionin tuomioistuimen ratkaisukäytäntöä erityisesti yhtä säilytetyn toimivallan alaa, koulutusta, koskevissa tapauksissa pyrkien havaitsemaan typologioita tuomioistuimen ratkaisutoiminnassa. Tutkimus noudattaa EU-valtiosääntöoikeuden metodologiaa. Keskeisenä lähdemateriaalina on siten käytetty unionituomioistuimen ratkaisukäytäntöä, joka heijastaa perustamissopimuksia tarkemmin unionin valtiosääntörakennetta. Oikeuskäytännön analyysi ja tulkinta on suoritettu peilaten sitä vasten unionin kehitystä markkinaorientoituneesta organisaatiosta yleismaailmalliseksi poliittiseksi unioniksi. Tutkielmani loppupäätelmä on, että jäsenvaltiot ovat tietyissä rajoissa hyväksyneet unionituomioistuimen kehittämän doktriinin, ja unionituomioistuin on siten saanut aikaan tosiasiallisen muutoksen EU:n valtiosääntörakenteessa. Retained powers -doktriini on omiaan syventämään eurooppalaista integraatiota ja nostaa kysymyksiä toimivallanjaon merkityksestä unionioikeudessa, perustamissopimusten kyvystä heijastaa unionin valtiosääntörakennetta sekä unionituomioistuimen toiminnan poliittisesta luonteesta.

The effect of age on the structure and compostition of the cell walls of Choanephora cucurbitarum

The effect of age on the structure and composition of isolated and purified cell walls from cultures of Choanephora cucurbitarum was investigated by microchemical analyses, visible and infrared spectrophotometry, x-ray diffractometry and electron microscopy. Qualitative evaluation revealed the presence of lipids, proteins, neutral sugars, strong alkali soluble sugars, chitin, chitosan and uronic acids in the cell walls of both the 1 and 7 day old cultures. As the mycelium aged, there was a slight but statistically significant increase in the protein content, and a pronounced rise in the chitin and neutral sugar constituents of the cell walls. Conversely, the decrease in the chitosan content during this period had the net effect of altering the chitin: chitosan ratio from near unity in the younger cultures, to a 2:1 ratio in the 7 day old cell wall samples. Glutaraldehyde-osmium fixed thin sections of the 1 day old vegetative hyphae of £. curbitarum revealed the presence of a monolayered cell wall, which upon aging became bilayered. Replicas of acid hydrolysed cell walls demonstrated that both the 1 and 7 day old samples possessed an outer layer which was composed of finely granular amorphous material and randomly distributed microfibrils. The deposition of an inner secondary layer composed of parallel oriented microfibrils in the older hypha was correlated with an increase in the chitin content in the cell wall. The significance of these results with respect to the intimate relationship between composition and structure is discussed.

The effect of 17-B estradiol therapy on bone mineral density and structure of alveolar bone in the ovariectomized rat model of postmenopausal osteoporosis

The ovariectomized (OVX) rat, a preclinical model for studying postmenopausal bone loss, may also be used to study differences in alveolar bone (AB). The objectives of this study were to quantify the differences in AB following estrogen replacement therapy (ERT), and to investigate the relationship between AB structure and density, and trabecular bone at the femoral neck (FN) and third lumbar vertebral body (LB3). Estrogen treated rats had a higher bone volume fraction (BV/TV) at the AB region (9.8% P < 0.0001), FN (12% P < 0.0001), and LB3 (11.5% P < 0.0001) compared to the OVX group. BV/TV of the AB was positively correlated with the BV/TV at the FN (r = 0.69 P < 0.0001) and the LB3 (r = 0.75 P < 0.0001). The trabecular number (Tb.N), trabecular separation (Tb.Sp), and structure model index (SMI) were also positively correlated (P < 0.05) between the AB and FN (r = 0.42, 0.49, and 0.73, respectfully) and between the AB and LB3 (r = 0.44, 0.63, and 0.69, respectfully). Given the capacity of AB to respond to ERT, future preclinical drug/nutritional intervention studies aimed at improving skeletal health should include the AB as a region of interest (ROI).

Investigation of the structure and dynamics of the centromeric epigenetic mark

Le centromère est le site chromosomal où le kinetochore se forme, afin d’assurer une ségrégation fidèles des chromosomes et ainsi maintenir la ploïdie appropriée lors de la mitose. L’identité du centromere est héritée par un mécanisme épigénétique impliquant une variante de l’histone H3 nommée centromere protein-A (CENP-A), qui remplace l’histone H3 au niveau de la chromatine du centromère. Des erreurs de propagation de la chromatine du centromère peuvent mener à des problèmes de ségrégation des chromosomes, pouvant entraîner l’aneuploïdie, un phénomène fréquemment observé dans le cancer. De plus, une expression non-régulée de CENP-A a aussi été rapportée dans différentes tumeurs humaines. Ainsi, plusieurs études ont cherchées à élucider la structure et le rôle de la chromatine contenant CENP-A dans des cellules en prolifération. Toutefois, la nature moléculaire de CENP-A en tant que marqueur épigénétique ainsi que ces dynamiques à l'extérieur du cycle cellulaire demeurent des sujets débat. Dans cette thèse, une nouvelle méthode de comptage de molécules uniques à l'aide de la microscopie à réflexion totale interne de la fluorescence (TIRF) sera décrite, puis exploitée afin d'élucider la composition moléculaire des nucléosomes contenant CENP-A, extraits de cellules en prolifération. Nous démontrons que les nucléosomes contenant CENP-A marquent les centromères humains de façon épigénétique à travers le cycle cellulaire. De plus, nos données démontrent que la forme prénucléosomale de CENP-A, en association avec la protéine chaperon HJURP existe sous forme de monomère et de dimère, ce qui reflète une étape intermédiaire de l'assemblage de nucléosomes contenant CENP-A. Ensuite, des analyses quantitatives de centromères lors de différenciation myogénique, et dans différents tissus adultes révèlent des changements globaux qui maintiennent la marque épigénétique dans une forme inactive suite à la différentiation terminale. Ces changements incluent une réduction du nombre de points focaux de CENP-A, un réarrangement des points dans le noyau, ainsi qu'une réduction importante de la quantité de CENP-A. De plus, nous démontrons que lorsqu'une dédifférenciation cellulaire est induite puis le cycle cellulaire ré-entamé, le phénotype "différencié" décrit ci-haut est récupéré, et les centromères reprennent leur phénotype "prolifératif". En somme, cet oeuvre décrit la composition structurale sous-jacente à l'identité épigénétique des centromères de cellules humaines lors du cycle cellulaire, et met en lumière le rôle de CENP-A à l'extérieur du cycle cellulaire.

Dynamic and Thermodynamic Structure of the Monsoon Boundary Layer over India

The objective of the study is to examine the dynamic and thermodynamic structure and the variations that occur in the surface layer during the pre-monsoon, onset and post-monsoon periods over the Indian region. The variations caused during the occurrence of micro and mesoscale systems, structure and variation in the marine boundary layer over the Indian region is also investigated. The drag coefficient computed indirectly also shows variation during various seasons. The thermodynamic structure of the atmosphere shows variation during the various seasons. The onset monsoon causes lowering of the Lifting Condensation Levels. The outcome of the study is expected to provide a better understanding of the structure and variations in the boundary layer over India, which is useful for many applications especially for numerical modeling studies.

Geophysical Constraints on Structure and Tectonics of the Eastern Arabian Sea and the Adjoining West Coast of India with Special Reference to the Kerala Basin

The main objective of the present study is to model the gravity fields in terms of lithospheric structure below the western continental margin of India (WCMI) identify zones of crustal mass anomalies and attempt to infer the location of Ocean Continent transition in the Arabian Sea. In this study, the area starting from the western shield margin to the region covering the deep oceanic parts of the Arabian Sea which is bounded by Carlsberg and Cerg and Central Indian ridges in the south, eastern part of the Indus Cone in the west and falling between 630E and 800E longitudes, and 50N - 240N latitudes has been considered. The vast amount of seismic reflection and refraction data in the form of crustal velocities, basement configuration and crustal thicknesses available for the west coast as well as the eastern Arabian Sea has been utilized for this purpose

Studies on the Composition and Structure of Palm Oil Glycerides

During the last couple of decades, the oil palm has emerged as the second largest source of edible oil in the world. Recently oil palm has been introduced commercially in India to augment edible oil supply in the country. Currently, about 10,000 hectares are under oil palm cultivation in India, and it is envisaged to cover about 6 lakh hectares in the coming years. Though oil palm is a major commercial oil crop, not much basic information on the lipids of the fruit (the source of palm oil) is available even where oil palm is cultivated in a very large scale. Being a new crop to India, it is of paramount importance to understand the basic chemistry/biochemistry of the lipids, which in turn, may find practical applications in the area of processing and product development. The present investigation entitled "Studies on the Composition and Structure of Palm Oil Glycerides" was designed with a view to elucidate the lipid composition and structure under conditions such as fruit development and processing.