Knockdown of zebrafish crim1 results in a bent tail phenotype with defects in somite and vascular development

The Crim1 gene encodes a transmembrane protein containing six cysteine-rich repeats similar to those found in the BMP antagonist, chordin (chd). To investigate its physiological role, zebrafish crim1 was cloned and shown to be both maternally and zygotically expressed during zebrafish development in sites including the vasculature, intermediate cell mass. notochord, and otic vesicle. Bent or hooked tails with U-shaped somites were observed in 85% of morphants from 12 hpf. This was accompanied by a loss of muscle pioneer cells. While morpholino knockdown of crim1 showed some evidence of ventralisation, including expansion of the intermediate cell mass (ICM), reduction in head size bent tails and disruption to the somites and notochord, this did not mimic the classically ventralised phenotype, as assessed by the pattern of expression of the dorsal markers chordin, otx2 and the ventral markers eve1, pax2.1, tall and gata1 between 75% epiboly and six-somites. From 24 hpf, morphants displayed an expansion of the ventral mesoderm-derived ICM, as evidenced by expansion of tall. Imo2 and crim1 itself. Analysis of the crim1 morphant phenotype in Tg(fli:EGFP) fish showed a clear reduction in the endothelial cells forming the intersegmental vessels and a loss of the dorsal longitudinal anastomotic vessel (DLAV). Hence, the primary role of zebrafish crim1 is likely to be the regulation of somitic and vascular development. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

Glacial cold-water coral: ages, isotope concentrations and ratios

A set of 40 Uranium-series datings obtained on the reef-forming scleractinian cold-water corals Lophelia pertusa and Madrepora oculata revealed that during the past 400 kyr their occurrence in the Gulf of Cádiz (GoC) was almost exclusively restricted to glacial periods. This result strengthens the outcomes of former studies that coral growth in the temperate NE Atlantic encompassing the French, Iberian and Moroccan margins dominated during glacial periods, whereas in the higher latitudes (Irish and Norwegian margins) extended coral growth prevailed during interglacial periods. Thus it appears that the biogeographical limits for sustained cold-water coral growth along the NE Atlantic margin are strongly related to climate change. By focussing on the last glacial-interglacial cycle, this study shows that palaeo-productivity was increased during the last glacial. This was likely driven by the fertilisation effect of an increased input of aeolian dust and locally intensified upwelling. After the Younger Dryas cold event, the input of aeolian dust and productivity significantly decreased concurrent with an increase in water temperatures in the GoC. This primarily resulted in reduced food availability and caused a widespread demise of the formerly thriving coral ecosystems. Moreover, these climate induced changes most likely caused a latitudinal shift of areas withoptimum coral growth conditions towards the northern NE Atlantic where more suitable environmental conditions established with the onset of the Holocene.

(Table 2) Radiocarbon dating on cold-water corals collected in the Mediterranean Sea

This study presents newly obtained coral ages of the cold-water corals Lophelia pertusa and Madrepora oculata collected in the Alboran Sea and the Strait of Sicily (Urania Bank). These data were combined with all available Mediterranean Lophelia and Madrepora ages compiled from literature to conduct a basin-wide assessment of the spatial and temporal occurrence of these prominent framework-forming scleractinian species in the Mediterranean realm and to unravel the palaeo-environmental conditions that controlled their proliferation or decline. For the first time special focus was placed on a closer examination of potential differences occurring between the eastern and western Mediterranean sub-basins. Our results clearly demonstrate that cold-water corals occurred sparsely in the entire Mediterranean during the last glacial before becoming abundant during the Bølling-Allerød warm interval, pointing to a basin-wide, almost concurrent onset in (re-)colonisation after ~13.5 ka. This time coincides with a peak in meltwater discharge originating from the northern Mediterranean borderlands which caused a major reorganisation of the Mediterranean thermohaline circulation. During the Younger Dryas and Holocene, some striking differences in coral proliferation were identified between the sub-basins such as periods of highly prolific coral growth in the eastern Mediterranean Sea during the Younger Dryas and in the western basin during the Early Holocene, whereas a temporary pronounced coral decline during the Younger Dryas was exclusively affecting coral sites in the Alboran Sea. Comparison with environmental and oceanographic data revealed that the proliferation of the Mediterranean corals is linked with enhanced productivity conditions. Moreover, corals thrived in intermediate depths and showed a close relationship with intermediate water mass circulation in the Mediterranean sub-basins. For instance, reduced Levantine Intermediate Water formation hampered coral growth in the eastern Mediterranean Sea during sapropel S1 event as reduced Winter Intermediate Water formation did in the westernmost part of the Mediterranean (Alboran Sea) during the Mid-Holocene. Overall, this study clearly demonstrates the importance to consider region-specific environmental changes as well as species-specific environmental preferences in interpreting coral chronologies. Moreover, it highlights that the occurrence or decline of cold-water corals is not controlled by one key parameter but rather by a complex interplay of various environmental variables.

Short-term metabolic and growth responses of the cold-water coral lophelia pertusa to ocean acidification

Cold-water corals are amongst the most three-dimensionally complex deep-sea habitats known and are associated with high local biodiversity. Despite their importance as ecosystem engineers, little is known about how these organisms will respond to projected ocean acidification. Since preindustrial times, average ocean pH has already decreased from 8.2 to ~ 8.1. Predicted CO2 emissions will decrease this by up to another 0.3 pH units by the end of the century. This decrease in pH may have a wide range of impacts upon marine life, and in particular upon calcifiers such as cold-water corals. Lophelia pertusa is the most widespread cold-water coral (CWC) species, frequently found in the North Atlantic. Data here relate to a short term data set (21 days) on metabolism and net calcification rates of freshly collected L. pertusa from Mingulay Reef Complex, Scotland. These data from freshly collected L. pertusa from the Mingulay Reef Complex will help define the impact of ocean acidification upon the growth, physiology and structural integrity of this key reef framework forming species.

Ionisation and discharge in cloud-forming atmospheres of brown dwarfs and extrasolar planets

Brown dwarfs and giant gas extrasolar planets have cold atmospheres with rich chemical compositions from which mineral cloud particles form. Their properties, like particle sizes and material composition, vary with height, and the mineral cloud particles are charged due to triboelectric processes in such dynamic atmospheres. The dynamics of the atmospheric gas is driven by the irradiating host star and/or by the rotation of the objects that changes during its lifetime. Thermal gas ionisation in these ultra-cool but dense atmospheres allows electrostatic interactions and magnetic coupling of a substantial atmosphere volume. Combined with a strong magnetic field , a chromosphere and aurorae might form as suggested by radio and x-ray observations of brown dwarfs. Non-equilibrium processes like cosmic ray ionisation and discharge processes in clouds will increase the local pool of free electrons in the gas. Cosmic rays and lighting discharges also alter the composition of the local atmospheric gas such that tracer molecules might be identified. Cosmic rays affect the atmosphere through air showers in a certain volume which was modelled with a 3D Monte Carlo radiative transfer code to be able to visualise their spacial extent. Given a certain degree of thermal ionisation of the atmospheric gas, we suggest that electron attachment to charge mineral cloud particles is too inefficient to cause an electrostatic disruption of the cloud particles. Cloud particles will therefore not be destroyed by Coulomb explosion for the local temperature in the collisional dominated brown dwarf and giant gas planet atmospheres. However, the cloud particles are destroyed electrostatically in regions with strong gas ionisation. The potential size of such cloud holes would, however, be too small and might occur too far inside the cloud to mimic the effect of, e.g. magnetic field induced star spots.

Modeling chemistry in massive star forming regions with internal PDRs

Over the past decades star formation has been a very attractive field because knowledge of star formation leads to a better understanding of the formation of planets and thus of our solar system but also of the evolution of galaxies. Conditions leading to the formation of high-mass stars are still under investigation but an evolutionary scenario has been proposed: As a cold pre-stellar core collapses under gravitational force, the medium warms up until it reaches a temperature of 100 K and enters the hot molecular core (HMC) phase. The forming central proto-star accretes materials, increasing its mass and luminosity and eventually it becomes sufficiently evolved to emit UV photons which irradiate the surrounding environment forming a hyper compact (HC) and then a ultracompact (UC) HII region. At this stage, a very dense and very thin internal photon-dominated region (PDR) forms between the HII region and the molecular core. Information on the chemistry allows to trace the physical processes occurring in these different phases of star formation. Formation and destruction routes of molecules are influenced by the environment as reaction rates depend on the temperature and radiation field. Therefore, chemistry also allows the determination of the evolutionary stage of astrophysical objects through the use of chemical models including the time evolution of the temperature and radiation field. Because HMCs host a very rich chemistry with high abundances of complex organic molecules (COMs), several astrochemical models have been developed to study the gas phase chemistry as well as grain chemistry in these regions. In addition to HMCs models, models of PDRs have also been developed to study in particular photo-chemistry. So far, few studies have investigated internal PDRs and only in the presence of outflows cavities. Thus, these unique regions around HC/UCHII regions remain to be examined thoroughly. My PhD thesis focuses on the spatio-temporal chemical evolution in HC/UC HII regions with internal PDRs as well as in HMCs. The purpose of this study is first to understand the impact and effects of the radiation field, usually very strong in these regions, on the chemistry. Secondly, the goal is to study the emission of various tracers of HC/UCHII regions and compare it with HMCs models, where the UV radiation field does not impact the region as it is immediately attenuated by the medium. Ultimately we want to determine the age of a given region using chemistry in combination with radiative transfer.

Novel biomaterials for regeneration of periapical bone defects in endodontic and implant therapy

Endodontic-related periapical bone defects are a common occurrence in the global populations. Considering the number of root canal treatments performed annually, new strategies and new biomaterials for the management of these bone defects will be important and highlight the need for continued research and development in endodontic field. The present PhD thesis have several objectives and is divided into two main sections: one focused on in vitro and laboratory research and the other on clinical in vivo investigations. The first part, focused on laboratory and in vitro research, investigated 2 main topics: • the microbial communities of apical periodontitis to evaluate the predominant bacterial using 16sr DNA-targeted Nanopore sequencing; • the physical-chemical properties of innovative premixed calcium-silicate based bioceramic sealers for endodontic therapy; The second part, focused on in vivo clinical studies, investigated 2 main topics: • the clinical application of premixed calcium-silicate-based sealers. Ethical committee approval was obtained in 2 separate in vivo studies. The first one is a prospective cohort study with a two-year follow-up where the test group was compared with a control group (considered the gold standard). The second is a pilot prospective cohort study with a 12-month follow-up which set the foundation for a subsequent randomized investigation. Thanks to these investigations, we validated a new technique that innovatively associates a warm obturation technique with calcium-silicate-based sealers. Historically, these sealers were only used with cold techniques. This investigation highlights the possibility for wider utilization and improvements in endodontic techniques. • The outcome of 2 different types of implants characterized by different surface treatments and placed with different techniques. The marginal bone level and periodontal parameters were evaluated with a follow-up of 4 and 10 years. This Ph.D thesis is based on a compilation of published papers I have done during my three-year PhD program.

Leaf-, Panel- And Latex-expressed Sequenced Tags From The Rubber Tree (hevea Brasiliensis) Under Cold-stressed And Suboptimal Growing Conditions: The Development Of Gene-targeted Functional Markers For Stress Response.

Hevea brasiliensis is a native species of the Amazon Basin of South America and the primary source of natural rubber worldwide. Due to the occurrence of South American Leaf Blight disease in this area, rubber plantations have been extended to suboptimal regions. Rubber tree breeding is time-consuming and expensive, but molecular markers can serve as a tool for early evaluation, thus reducing time and costs. In this work, we constructed six different cDNA libraries with the aim of developing gene-targeted molecular markers for the rubber tree. A total of 8,263 reads were assembled, generating 5,025 unigenes that were analyzed; 912 expressed sequence tags (ESTs) represented new transcripts, and two sequences were highly up-regulated by cold stress. These unigenes were scanned for microsatellite (SSR) regions and single nucleotide polymorphisms (SNPs). In total, 169 novel EST-SSR markers were developed; 138 loci were polymorphic in the rubber tree, and 98 % presented transferability to six other Hevea species. Locus duplication was observed in H. brasiliensis and other species. Additionally, 43 SNP markers in 13 sequences that showed similarity to proteins involved in stress response, latex biosynthesis and developmental processes were characterized. cDNA libraries are a rich source of SSR and SNP markers and enable the identification of new transcripts. The new markers developed here will be a valuable resource for linkage mapping, QTL identification and other studies in the rubber tree and can also be used to evaluate the genetic variability of other Hevea species, which are valuable assets in rubber tree breeding.

O esporte paralímpico na formação do profissional em educação física : percepção de professores e acadêmicos = The paralympic sport in physical education professional forming: perceptions of teachers and students

Universidade Estadual de Campinas . Faculdade de Educação Física

Digital subtraction radiographic analysis of the combination of bioabsorbable membrane and bovine morphogenetic protein pool in human periodontal infrabony defects

OBJECTIVES: This study assessed the bone density gain and its relationship with the periodontal clinical parameters in a case series of a regenerative therapy procedure. MATERIAL AND METHODS: Using a split-mouth study design, 10 pairs of infrabony defects from 15 patients were treated with a pool of bovine bone morphogenetic proteins associated with collagen membrane (test sites) or collagen membrane only (control sites). The periodontal healing was clinically and radiographically monitored for six months. Standardized pre-surgical and 6-month postoperative radiographs were digitized for digital subtraction analysis, which showed relative bone density gain in both groups of 0.034 ± 0.423 and 0.105 ± 0.423 in the test and control group, respectively (p>0.05). RESULTS: As regards the area size of bone density change, the influence of the therapy was detected in 2.5 mm² in the test group and 2 mm² in the control group (p>0.05). Additionally, no correlation was observed between the favorable clinical results and the bone density gain measured by digital subtraction radiography (p>0.05). CONCLUSIONS: The findings of this study suggest that the clinical benefit of the regenerative therapy observed did not come with significant bone density gains. Long-term evaluation may lead to a different conclusions.

Repair of critical-size defects with autogenous periosteum-derived cells combined with bovine anorganic apatite/collagen: an experimental study in rat calvaria

The aim of this study was to evaluate the bone repair using autogenous periosteum-derived cells (PDC) and bovine anorganic apatite and collagen (HA-COL). PDC from Wistar rats (n=10) were seeded on HA-COL discs and subjected to osteoinduction during 6 days. Critical-size defects in rat calvarias were treated with blood clot (G1), autogenous bone (G2), HA-COL (G3) and HA-COL combined with PDC (G4) (n=40), and then analyzed 1 and 3 months after surgeries. Radiographic analysis exhibited no significant temporal change. G1 and G2 had discrete new marginal bone, but the radiopacity of graft materials in G2, G3 and G4 impaired the detection of osteogenesis. At 3 months, histopathological analysis showed the presence of ossification islets in G1, which was more evident in G2, homogeneous new bone around HA-COL in G3 and heterogeneous new bone around HA-COL in G4 in addition to moderate presence of foreign body cells in G3 and G4. Histomorphometric analysis showed no change in the volume density of xenograft (p>0.05) and bone volume density in G2 was twice greater than in G1 and G4 after 3 months (p<0.05), but similar to G3. The PDC did not increase bone formation in vivo, although the biomaterial alone showed biocompatibility and osteoconduction capacity.

Quality of sausage elaborated using minced Nile Tilapia submmitted to cold storage

Filleting yield of Nile tilapia Oreochromis niloticus (L.) is low (30%) and generates large amount of wastes that may turn into environmental and economic problem. However, these wastes can be used for the extraction of minced fish (MF) which can be used in the preparation of sausages. The objective of this study was to assess the quality of sausages prepared with 0, 20, 40, 60, 80 and 100% of MF from Nile tilapia filleting waste during storage at 0±0.3ºC. Alterations in the instrumental color (L*, a* and b*), lipid oxidation (TBARS), total volatile nitrogenous bases (TVB-N), pH, microbiological condition (pathogenic bacteria and aerobic psychrotrophic bacteria), and sensory attributes (color, odor, flavor, texture and overall acceptability) were evaluated for up to 40 days. The addition of MF to sausages increased TBARS values and decreases TVB-N, L*, a* and b* values. Acceptability of color attribute decreased with increasing MF; best flavor, texture and overall acceptability scores were registered for sausages containing 40 and 60% MF; best odor was registered for 100% MF. Pathogenic microorganisms were not detected, but decrease in pH and proliferation of aerobic psychrotrophic bacteria which, however, did not compromise sensory evaluation of sausages were registered throughout storage. Sausages prepared with MF from tilapia filleting waste have a shelf-life of 40 days when stored at 0±0.3ºC, and the maximum recommended MF inclusion to maintain good sensory quality is 60%.

Effect of cold fronts on the benthic macrofauna of exposed sandy beaches with contrasting morphodynamics

The aim of this study was to determine the short-term environmental changes caused by the simultaneous passage of a high energy event on two sandy beaches with different morphodynamic states and their influence on the richness, abundance and distribution of the benthic macrofauna. Two microtidal exposed sandy beaches with contrasting morphodynamics were simultaneously sampled before, during and after the passage of two cold fronts in Santa Catarina. The reflective beach showed a higher susceptibility to the increase in wave energy produced by the passage of cold fronts and was characterized by rapid and intense erosive processes in addition to a capacity for rapid restoration of the beach profile. As regards the dissipative beach, erosive processes operated more slowly and progressively, and it was characterized further by a reduced capacity for the recovery of its sub-aerial profile. Although the intensity of the environmental changes was distinct as between the morphodynamic extremes, changes in the composition, richness and abundance of macrobenthos induced by cold fronts were not evident for either of the beaches studied. On the other hand, alterations in the distribution pattern of the macrofauna were observed on the two beaches and were related to variations in sea level, position of the swash zone and moisture gradient, suggesting that short-term accommodations in the spatial structure of the macrobenthos occur in response to changes in environmental conditions in accordance with the temporal dynamics characteristic of each morphodynamic state.

Memantine prevents cardiomyocytes nuclear size reduction in the left ventricle of rats exposed to cold stress

OBJECTIVES: Memantine is an N-methyl-d-aspartate (NMDA) glutamate receptor antagonist used to treat Alzheimer's disease. Previous studies have suggested that receptor blockers act as neuroprotective agents; however, no study has specifically investigated the impact that these drugs have on the heart. We sought to evaluate the effects of memantine on nuclear size reduction in cardiac cells exposed to cold stress. METHOD: We used male EPM-Wistar rats (n=40) divided into 4 groups: 1) Matched control (CON); 2) Memantine-treated rats (MEM); 3) Rats undergoing induced hypothermia (IH) and 4) Rats undergoing induced hypothermia that were also treated with memantine (IHM). Animals in the MEM and IHM groups were treated by oral gavage administration of 20 mg/kg/day memantine over an eight-day period. Animals in the IH and IHM groups were submitted to 4 hours of hypothermia in a controlled environment with a temperature of - 8ºC on the last day of the study. RESULTS: The MEM group had the largest cardiomyocyte nuclear size (151 ± 3.5 μm³ vs. CON: 142 ± 2.3 μm³; p<0.05), while the IH group had the smallest mean value of nuclear size. The nuclear size of the IHM group was preserved (125 ± 2.9 μm³) compared to the IH group (108 ± 1.7 μm³; p<0.05). CONCLUSION: Memantine prevented the nuclear size reduction of cardiomyocytes in rats exposed to cold stress.