THE IMPORTANCE OF MHC-INDEPENDENT NATURAL KILLER CELL RECEPTORS IN THE IMMUNOREGULATION OF MURINE PREGNANCY

Numerous leukocyte populations are essential for pregnancy success. Uterine natural killer (uNK) cells are chief amongst these leukocytes and represent a unique lineage with limited cytotoxicity but abundant angiokine production. They possess a distinct phenotype of activating and inhibitory receptors that recognize major histocompatibility complex (MHC) molecules, such as the killer immunoglobulin like receptors (KIRs; mouse Ly49), and MHC-independent activating receptors, including the aryl hydrocarbon receptor (AHR) and natural cytotoxicity receptor 1 (NCR1). While the roles of MHC-dependent receptors are widely addressed in pregnancy, MHC-independent receptors are relatively unstudied. This thesis investigated the roles of MHC-independent receptors in promotion of mouse pregnancy and characterized early leukocyte interactions in the presence and absence of NCR1. It was hypothesized that loss of MHC-independent receptors impairs uNK cell development resulting in aberrations in leukocyte function and decidual vasculature. Implantation sites from Ahr-/- and Ncr1Gfp/Gfp mice were assessed using whole mount in situ immunohistochemistry (WM-IHC) and histochemical techniques. Leukocyte interactions identified during preliminary WM-IHC studies were confirmed as immune synapses. The novel identification of immune synapses in early mouse pregnancy compelled further examination of leukocyte conjugates in wildtype C57BL/6 and Ncr1Gfp/Gfp mice. In Ahr-/- and Ncr1Gfp/Gfp mice, receptor loss resulted in reduced uNK cell diameters, impaired decidual vasculature, and failures in spiral artery remodeling. Ahr-/- mice had severe fertility deficits whereas Ncr1Gfp/Gfp mice had increased fetal resorption indicating differing receptor requirements in pregnancy success. NCR1 loss primarily affected uNK cell maturation and function as identified by alterations in granule ultrastructure, lytic protein expression, and angiokine production. Leukocyte conjugates were frequent in early C57BL/6 decidua basalis and included uNK cells conjugating first with antigen presenting cells and then with T cells. Overall conjugate formation was reduced in the absence of NCR1, but specific uNK cell conjugations were unaffected by receptor loss. While KIR-MHC interactions are associated with numerous pregnancy complications in humans, the role of other uNK cell receptors are not well characterized. These results illustrate the importance of MHC-independent receptors in uNK cell activation during early pregnancy in mice and encourage further studies of pregnancy complications that may occur independently of maternal KIR-MHC contributions.

Neutrophilic Cathepsin C Is Maturated by a Multistep Proteolytic Process and Secreted by Activated Cells during Inflammatory Lung Diseases

The cysteine protease cathepsin C (CatC) activates granule-associated proinflammatory serine proteases in hematopoietic precursor cells. Its early inhibition in the bone marrow is regarded as a new therapeutic strategy for treating proteolysis-driven chronic inflammatory diseases, but its complete inhibition is elusive in vivo Controlling the activity of CatC may be achieved by directly inhibiting its activity with a specific inhibitor or/and by preventing its maturation. We have investigated immunochemically and kinetically the occurrence of CatC and its proform in human hematopoietic precursor cells and in differentiated mature immune cells in lung secretions. The maturation of proCatC obeys a multistep mechanism that can be entirely managed by CatS in neutrophilic precursor cells. CatS inhibition by a cell-permeable inhibitor abrogated the release of the heavy and light chains from proCatC and blocked ∼80% of CatC activity. Under these conditions the activity of neutrophil serine proteases, however, was not abolished in precursor cell cultures. In patients with neutrophilic lung inflammation, mature CatC is found in large amounts in sputa. It is secreted by activated neutrophils as confirmed through lipopolysaccharide administration in a nonhuman primate model. CatS inhibitors currently in clinical trials are expected to decrease the activity of neutrophilic CatC without affecting those of elastase-like serine proteases.

Investigation on reproduction process in two species of crab Eriphia sebana and Ocypode saratan in the intertidal zone of Chabahar area

The present study with headline investigation on reproduction in two species of Crab Eriphia sebana and Ocypode saratan was carried out in the intertidal zone of Chabahar in thirteen month from December 2004 to December 2005. Checked samplings have been taken, 45 number Crab monthly from any four stations by manual or use trap. During this study the following subjects were measured: Temperature range and salinity, Measurable coast granule, Determination of sex ratio, Relations carapace width with carapace length, Carapace width with total body weight, Gonad weight, gonadosomatic index, condition factor, gastrosomatic index, investigation content in stomach, LM50, Growth parameters, plenty distribution length and width and gonad weight and total body weight. Studied on measurable coast granule were expressed that Ocypode saratan in Desalination station, were nest in soils equable sand and this quantity were confirmed in Pozm station. Sex ratio were assign in desalination area and Pozm M: F 0/44:0/56 and in Tiss and Chabahar M: F 0/45:0/55. Carapace length and carapace width (cm) and body weight (g) Furthest were designated in Ocypode saratan within carapace width sequential: female: 5/42-6/15-105/13 and male: 5/53-6/25-108/91 and in Eriphia sebana within Tiss area sequential: female: 5/12-5/94-110/21 and male 5/14-60/01-114/37. Have been linear relationship between carapace length and carapace width and equaled CW = a CL + b. Weight growth in two species were be modal and equaled BW= aCLb and increased Crab weight by built up carapace width. Maximum gonad weight in Ocypode saratan within desalination area in female have been outcome 3/39 and in male 0/84g and in Eriphia sebana extreme within Tiss during may in female were be 4/18 and in male 1/1g. Stomach content in Eriphia sebana were involved a black until half-purplish liquid and yellowish in Ocypode saratan. Stomach contents identifiable were being in four groups: Molluscoid, Crustacean, Plankton and Fish. Carapace width during the first year of maturation have been LM50:3/77 in Desalination area and LM50:3/92 in Pozm for Ocypode saratan and LM50:4/26 in Tiss and LM50:4/62 in Chabahar. Ability spawning in Eriphia sebana within Tiss has been CW=4/17cm and in Ocypode saratan within Desalination area CW=4/23cm. Maximun value of Loo for Eriphia sebana was equal 59/67 and growth factor K=0/68 within Tiss and Loo =61/64 , K=0/65 for Ocypode saratan within Desalination area. Maximun GSI and GI have been within Desalination area and Tiss and minimum within Pozm and Chabahar. The maturity stages of two species were classifed into six stages. Review on GSI, CF have been showed that relation with temperature and salinity and definer in two species have been spawned in two period that Maximun in spring premier than autumn.

3D Porous Architecture of Stacks of β-TCP Granules Compared with That of Trabecular Bone: A microCT, Vector Analysis, and Compression Study

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Structural and ultrastructural characteristics of interrenal gland and chromaffin cell of matrinxa, Brycon cephalus Gunther 1869 (Teleostei-Characidae)

This work presents the structure and ultrastructure of the interrenal gland and chromaffin cells, as well as the morphology of the head kidney of Brycon cephalus, the head kidney is composed of fused bilateral lobes located anterior to the swim bladder and ventrolateral to the spinal column, the parenchyma revealed lympho-haematopoietic tissue, melano-macrophage centres, interrenal gland and chromaffin cells. The interrenal gland consisted of cords or strands of cells grouped around the posterior cardinal vein and their branches. Chromaffin cells are found in small groups, closely associated with the interrenal gland and/or under the endothelium of the posterior cardinal vein. So far, the ultrastructural analysis has revealed only one interrenal cell type which contained abundant smooth endoplasmic reticulum and numerous mitochondria with tubulo-vesicular cristae, characteristic of steroid-producing cells. Two types of chromaffin cells were observed. The first type was characterized by the presence of vesicles with round, strongly electron-dense granules, which were eccentrically located, Such cells were interpreted as noradrenaline cells, Meanwhile, cells which contained smaller vesicles and electron-lucent granules, with a small halo separating the granule from the vesicular limiting membrane, were identified as adrenaline cells.

Sexagem histológica e desempenho de Oreochromis niloticus testando diâmetros de ração de acordo com o aparato bucal

Estudou-se o tamanho da boca de larvas de tilápia-do-nilo e testou-se o efeito de diferentes granulometrias da ração sobre o ganho de peso, comprimento e sobrevivência das larvas aos 30 e 60 dias de arraçoamento. Avaliou-se também o método de sexagem por meio de microscopia de luz aos 35 dias de idade. A medida da boca das larvas apresentou valores médios de 918,2±152,9μm aos cinco dias de idade. de acordo com esse dado, testaram-se três granulometrias: 0,25, 0,35 e 0,50mm. Aos 30 e aos 60 dias de arraçoamento, 10% das larvas foram medidas, pesadas e contadas para cálculo da taxa de sobrevivência. O tamanho dos grânulos testados não afetou o desempenho das larvas de tilápia nilótica com alimentação iniciada aos cinco dias pós-eclosão. Quanto à sexagem histológica aos 35 dias de idade, as gônadas apresentaram-se, em sua maioria, indiferenciadas. Recomenda-se que essa análise deva ser realizada de acordo com o tamanho dos animais e não com a idade.

Structural and functional interaction between polyamine related molecules and biological membranes

Changes induced by PA on nucleic acid (NA) conformation and synthesis is proven to be a major reason for PA essentiality (1-3). However, PA interactions with other polyanions, for instance polyanionic membrane lipid bilayers and glyosaminoglycans have received less attention (3-4). The functional importance of these interactions still is an obscure but interesting area of cell and molecular biology, especially in mammalian cells for which specific PA transport systems are not fully characterized (5). In mammals, activity and turnover of the polyamine (PA) synthesis key enzyme is controlled by a set of proteins: Antizymes (OAZ1-3) and antizyme inhibitors (AZIN1 and 2). It is demonstrated that AOZ modulate polyamine uptake (6), and that PA transport to mitochondria is linked to the respiratory chain state and modulates mitochondrial permeability transition (7). Antizyme expression variants have been located in mitochondria, being proposed as a proapoptotic factor (7-8). AZIN 2 is only expressed in a reduced set of tissues that includes mast cells, where it is associated to mast cell granules membrane (9). This fact, together to the abnormalities observed in bone marrow derived mast cell granules when they are differentiated under restricted PA synthesis conditions (10 and unpublished results), point out to important roles of PA and their related proteins in structure and function of mast cell granules. We will also present novel biophysical results on tripartite interactions of PA that remark the interest of the characterization of PA interactions with lipid bilayers for biomedicine and biotechnology. Thus, the information reported in this paper integrates previously reported information with our still unpublished results, all indicating that PA and their related proteins also are important factors for structure and dynamics of biological membranes and their associated functions essential in human physiology; for instance, solute interchange with the environment (uptake and secretion), oxidative metabolism and apoptosis. The importance of these involved processes for human homeostasis claim for further research efforts. 1. Ruiz-Chica J, Medina MA, Sánchez-Jiménez F and Ramírez FJ (2001) Fourier Transform Raman study of the structural specificities on the interaction between DNA and biogenic polyamines. Biophysical J. 80:443-454. 2. Lightfoot HL, Hall J (2014) Endogenous polyamine function--the RNA perspective. Nucleic Acids Res. 42:11275-11290. 3. Igarashi K, Kashiwagi K (2010) Modulation of cellular function by polyamines. Int J Biochem Cell Biol. 42:39-51. 4. Finger S, Schwieger C, Arouri A, Kerth A, Blume A (2014) Interaction of linear polyamines with negatively charged phospholipids: the effect of polyamine charge distance. Biol Chem. 395:769-778. 5. Poulin R, Casero RA, Soulet D. (2012) Recent advances in the molecular biology of metazoan polyamine transport. Amino Acids. 42:711-723. 6. Kahana C (2009) Regulation of cellular polyamine levels and cellular proliferation by antizyme and antizyme inhibitor. Essays Biochem. 4:47-61. 7. Agostinelli E, Marques MP, Calheiros R, Gil FP, Tempera G, Viceconte N, Battaglia V, Grancara S, Toninello A (2010) Polyamines: fundamental characters in chemistry and biology. Amino Acids 38:393-403. 8. Liu GY, Liao YF, Hsu PC, Chang WH, Hsieh MC, Lin CY, Hour TC, Kao MC, Tsay GJ, Hung HC (2006) Antizyme, a natural ornithine decarboxylase inhibitor, induces apoptosis of haematopoietic cells through mitochondrial membrane depolarization and caspases' cascade. Apoptosis 11:1773-1788. 9. Kanerva K, Lappalainen J, Mäkitie LT, Virolainen S, Kovanen PT, Andersson LC (2009). Expression of antizyme inhibitor 2 in mast cells and role of polyamines as selective regulators of serotonin secretion. PLoS One 31:e6858. 10. García-Faroldi G, Rodríguez CE, Urdiales JL, Pérez-Pomares JM, Dávila JC, Pejler G, Sánchez-Jiménez F, Fajardo I (2010) Polyamines are present in mast cell secretory granules and are important for granule homeostasis. PLoS One 30:e15071.

Evaluation of double treated starches using thermal tools.

Starch is the main polysaccharide found in cereals, composed by amylose and amylopectin. Corn is the principal source of starches worldwide. Starches treatment, through physical, chemical and/or biological methods, can improve the applications range. Acid modification in alcoholic solution promotes minimally degradation in the granule. Ball mill is one physical method poorly explored. The aim was to treat the starches using HCl 0.5 mol L-1 for 1 hour in 100 ml of aqueous, ethanol or methanol solutions with subsequent ball milling processes. One sample was selected as native sample. The four others, one native sample and three acid modified samples, were treated by physical process with the oscillating ball mill. The DTG-60H equipment was used for the TG and DTA analysis. The TG curves showed three mass losses related to dehydration, decomposition and oxidation. The native sample without physical modification showed major resistance to total degradation. This occurs because the physical modification cleaves hydrogen bonds, leaving a weakened granule. The TGDTA results showed that the mass loss in the 2nd event was minor in the hydrolyzed samples compared with native samples. The acid modification can provide starch higher resistance to degradation up to 340 °C. These results showed that chemical and physical treatment changed the thermal behaviors of the starches.