Hypoxia Inhibits Hypertrophic Differentiation and Endochondral Ossification in Explanted Tibiae

Purpose: Hypertrophic differentiation of growth plate chondrocytes induces angiogenesis which alleviates hypoxia normally present in cartilage. In the current study, we aim to determine whether alleviation of hypoxia is merely a downstream effect of hypertrophic differentiation as previously described or whether alleviation of hypoxia and consequent changes in oxygen tension mediated signaling events also plays an active role in regulating the hypertrophic differentiation process itself. Materials and Methods: Fetal mouse tibiae (E17.5) explants were cultured up to 21 days under normoxic or hypoxic conditions (21% and 2.5% oxygen respectively). Tibiae were analyzed on growth kinetics, histology, gene expression and protein secretion. Results: The oxygen level had a strong influence on the development of explanted fetal tibiae. Compared to hypoxia, normoxia increased the length of the tibiae, length of the hypertrophic zone, calcification of the cartilage and mRNA levels of hypertrophic differentiation-related genes e.g. MMP9, MMP13, RUNX2, COL10A1 and ALPL. Compared to normoxia, hypoxia increased the size of the cartilaginous epiphysis, length of the resting zone, calcification of the bone and mRNA levels of hyaline cartilage-related genes e.g. ACAN, COL2A1 and SOX9. additionally, hypoxia enhanced the mRNA and protein expression of the secreted articular cartilage markers GREM1, FRZB and DKK1, which are able to inhibit hypertrophic differentiation. Conclusions: Collectively our data suggests that oxygen levels play an active role in the regulation of hypertrophic differentiation of hyaline chondrocytes. Normoxia stimulates hypertrophic differentiation evidenced by the expression of hypertrophic differentiation related genes. In contrast, hypoxia suppresses hypertrophic differentiation of chondrocytes, which might be at least partially explained by the induction of GREM1, FRZB and DKK1 expressio

Kinetics of TNF-alpha and IFN-gamma mRNA expression in islets and spleen of NOD mice

Insulin-dependent diabetes mellitus is caused by autoimmune destruction of pancreatic ß cells. Non-obese diabetic (NOD) mice spontaneously develop diabetes similar to the human disease. Cytokines produced by islet-infiltrating mononuclear cells may be directly cytotoxic and can be involved in islet destruction coordinated by CD4+ and CD8+ cells. We utilized a semiquantitative RT-PCR assay to analyze in vitro the mRNA expression of TNF-alpha and IFN-gamma cytokine genes in isolated islets (N = 100) and spleen cells (5 x 10(5) cells) from female NOD mice during the development of diabetes and from female CBA-j mice as a related control strain that does not develop diabetes. Cytokine mRNAs were measured at 2, 4, 8, 14 and 28 weeks of age from the onset of insulitis to the development of overt diabetes. An increase in IFN-gamma expression in islets was observed for females aged 28 weeks (149 ± 29 arbitrary units (AU), P<0.05, Student t-test) with advanced destructive insulitis when compared with CBA-j mice, while TNF-alpha was expressed in both NOD and CBA-j female islets at the same level at all ages studied. In contrast, TNF-alpha in spleen was expressed at higher levels in NOD females at 14 weeks (99 ± 8 AU, P<0.05) and 28 weeks (144 ± 17 AU, P<0.05) of age when compared to CBA-j mice. The data suggest that IFN-gamma and TNF-alpha expression in pancreatic islets of female NOD mice is associated with ß cell destruction and overt diabetes.1347135

Failure of sexing by developmental arrest of bovine embryos in vitro produced with H-Y antisera

Embriões bovinos produzidos in vitro, em estádio de mórula, foram cultivados em meio contendo anticorpos anti H-Y de alto título proveniente de ratos por 24h e, após este tempo, classificados em dois grupos: 1) embriões inibidos em estádio de mórula (classificados como machos) e 2) embriões que se desenvolveram e formaram a blastocele (classificados como fêmeas). O sexo de 311 embriões, distribuídos em três grupos de concentração dos anticorpos, 3%, 5% ou 7%, foi identificado pela reação em cadeia da polimerase. Não houve desvio da proporção entre machos e fêmeas (P>0,05) nos grupos em que se utilizaram os anticorpos anti H-Y, quando comparadas ao grupo-controle, sem adição de anticorpos anti H-Y. Diferentemente dos resultados obtidos utilizando-se embriões bovinos produzidos in vivo, a sexagem com anticorpos anti H-Y de alto título em embriões produzidos in vitro não propiciou sucesso.In vitro produced bovine embryos at morula stage were cultured in medium containing high titer of rat H-Y antisera for 24h. The embryos were classified in two groups: 1) embryos arrested at morula stage (classified as males); and 2) embryos that developed and formed a blastocoele (classified as female). The sex of 311 embryos, divided in three groups of concentration of H-Y antisera, 3%, 5% or 7%, was identified by polimerase chain reaction. The results showed no difference (P>0.05) on sexual deviation in groups in which the H-Y antisera was added, in relation to control group, in which no H-Y antisera was added. In contrast with results obtained with in vivo produced bovine embryos, the sexing of in vitro produced bovine embryos with high H-Y antisera titer did not succed.CNP

Prediction of apparent digestibility of hays from natural pastures of the Northeast region of Portugal

In the Northeast region of Portugal hays from natural pastures are traditionally used as the main feed during the period of drought and the evaluation of its nutritive value is essential. The aim of this study was to predict the apparent organic matter digestibility (OMD) of 21 hays from natural pastures based on chemical composition, pepsin-cellulase solubility of organic matter (OMS) and in vitro organic matter digestibility. Chemical composition of hays showed a wide range of values and acid detergent lignin (ADL) was the component that presented the highest variation (CV = 18.4%). The OMD varied between 516 and 658 g/kg OM. The best single OMD predictor was OMS (RSD = 2.8%, R2 = 0.52, P<0.001). Using multiple regression to predict OMD, the variables included in the model were OMS and ADL (OMD = 21.51 + 0.94 OMS – 0.83 ADL; RSD = 2.75%; R2 = 0.54; P<0.0001). According to principal component analysis (PCA) hays were divided in 3 groups and the multiple regression established for the larger group of hays (n=10) was: OMD = 4.13 + 0.85 OMS – 1.03 ADL; RSD = 2.13%, R2 = 0.77, P<0.0001. The OMS method was superior to chemical composition and to the in vitro rumen fluid method in predicting OMD of hays from natural pastures. Results from PCA suggested that it may be useful to group these hays according to its chemical composition to accurately predict OMD

Magnetic And Superconducting Properties Of Ru Sr2 Gd1.5 Ce0.5 Cu2 O10-δ Samples: Dependence On The Oxygen Content And Aging Effects

The magnetic and superconducting properties of Ru Sr2 Gd1.5 Ce0.5 Cu2 O10-δ polycrystalline samples with different oxygen-doping levels are presented. A strong suppression of the superconducting temperature (Tc), as well as a reduction in the superconducting fraction, occurs as the oxygen content is reduced by annealing the samples in oxygen-deprived atmospheres. Drastic changes in the electrical resistivity are observed above Tc, possibly associated with oxygen removal, mainly from grain boundaries. However, the magnetic ordering is relatively less affected by the changes in oxygen content of the samples. The spin-glass transition is enhanced and shifted to higher temperatures with the reduction in oxygen content. This could be correlated with an increase in the spin disorder and frustration for the oxygen-depleted samples. Also, the same oxygen-vacancy-induced disorder could explain the reduction in the fraction of the sample showing antiferromagnetic order. We also report significant changes in the measured properties of the samples as a function of time. © 2005 The American Physical Society.7113Felner, I., Asaf, U., Levi, Y., Millo, O., (1997) Phys. Rev. B, 55, p. 3374. , PRBMDO 0163-1829 10.1103/PhysRevB.55.R3374Bernhard, C., Tallon, J.L., Niedermayer, Ch., Blasius, Th., Golnik, A., Brücher, E., Kremer, R.K., Ansaldo, E.J., (1999) Phys. Rev. B, 59, p. 14099. , PRBMDO. 0163-1829. 10.1103/PhysRevB.59.14099Awana, V.P.S., Karppinen, M., Yamauchi, H., (2003) Studies of High Tc Superconductors, 46, p. 77. , edited by A. V. Narlikar (Nova Science Publishers, New YorkMatvejeff, M., Awana, V.P.S., Jang, L.-Y., Liu, R.S., Yamauchi, H., Karppinen, M., (2003) Physica C, 392-396, p. 87. , PHYCE6 0921-4534Cardoso, C.A., Araujo-Moreira, F.M., Awana, V.P.S., Kishan, H., Takayama-Muromachi, E., De Lima, O.F., (2004) Physica C, 405, p. 212. , PHYCE6 0921-4534Cardoso, C.A., Araujo-Moreira, F.M., Awana, V.P.S., Takayama-Muromachi, E., De Lima, O.F., Yamauchi, H., Karppinen, M., (2003) Phys. Rev. B, 67, p. 020407. , PRBMDO 0163-1829 10.1103/PhysRevB.67.020407Shi, L., Li, G., Fan, X.J., Feng, S.J., Li, X.-G., (2003) Physica C, 399, p. 69. , PHYCE6 0921-4534Felner, I., Asaf, U., Ritter, F., Klamut, P.W., Dabrowski, B., (2001) Physica C, 364-365, p. 368. , PHYCE6 0921-4534Felner, I., Asaf, U., Galstyan, E., (2002) Phys. Rev. B, 66, p. 024503. , PRBMDO 0163-1829 10.1103/PhysRevB.66.024503Felner, I., Asaf, U., Levi, Y., Millo, O., (2000) Physica C, 334, p. 141. , PHYCE6 0921-4534 10.1016/S0921-4534(00)00250-1Awana, V.P.S., Ansari, M.A., Gupta, A., Saxena, R.B., Kishan, H., Buddhikot, D., Malik, S.K., (2004) Phys. Rev. B, 70, p. 104520. , PRBMDO 0163-1829 10.1103/PhysRevB.70.104520Yoshizawa, H., Mitsuda, S., Aruga, H., Ito, A., (1987) Phys. Rev. Lett., 59, p. 2364. , PRLTAO 0031-9007 10.1103/PhysRevLett.59.2364Binder, K., Young, A.P., (1986) Rev. Mod. Phys., 58, p. 801. , RMPHAT 0034-6861 10.1103/RevModPhys.58.801Mydosh, J.A., (1993) Spin Glasses An Experimental Introduction, , Taylor & Francis, LondonFelner, I., Galstyan, E., Herber, R.H., Nowik, I., (2004) Phys. Rev. B, 70, p. 094504. , PRBMDO 0163-1829 10.1103/PhysRevB.70.094504Shengelaya, A., Khasanov, R., Eschenko, D.G., Felner, I., Asaf, U., Savić, I.M., Keller, H., Müller, K.A., (2004) Phys. Rev. B, 69, p. 024517. , PRBMDO. 0163-1829. 10.1103/PhysRevB.69.024517Xue, Y.Y., Cao, D.H., Lorenz, B., Chu, C.W., (2002) Phys. Rev. B, 65, p. 020511. , PRBMDO 0163-1829 10.1103/PhysRevB.65.02051

Structural And Magnetic Study Of Labacocu O5+δ

The structure and magnetic properties of the compound LaBaCuCo O5+δ have been studied for the non-stoichiometric oxygen concentration δ≈0.6. The structure is pseudo-cubic with a tripled perovskite unit cell. The crystal structure was determined by a combined Rietveld fit to neutron and synchrotron x-ray powder diffraction data in the orthorhombic Pmmm space group, with cell parameters a=3.9223 (3) Å, b=3.9360 (3) Å, c=11.7073 (8) Å, and V=180.74 (2) Å3 (room temperature). Antiferromagnetic ordering of Cu and Co magnetic moments is observed below 205 (4) K. The magnetic structure with cell aM =2a, bM =2b, and cM =2c, could be described with the Shubnikov space group Fmm m′. The magnetic moments of both equivalent Cu/Co sites were determined at 50 and 170 K to be 0.83 (3) μB and 0.58 (3) μB, respectively, consistent with one unpaired electron per atom. The fit of the intensities to a simple mean field magnetic model appeared to be insufficient to account for the variation of moments at temperatures close to TN while a three dimensional Heisenberg model could improve the fit. Susceptibility measurements between 4 and 350 K also show irreversibility below 150 K. The local environments of Cu and Co were studied by extended x-ray absorption fine structure spectroscopy at both absorption edges. Cu atoms adopt an elongated octahedral or square-based pyramidal oxygen environment which suggests mainly the presence of Cu(II) in the structure. Co adopts different local environments, depending on the electronic and spin states. © 2005 The American Physical Society.7114Er-Rakho, L., Michel, C., Lacorre, P., Raveau, B., (1988) J. 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High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo

Cell-based cartilage repair strategies such as matrix-induced autologous chondrocyte implantation (MACI) could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous matrix deposition and remodelling. To address this issue, we designed a micro-mould to enable controlled high-throughput formation of micro-aggregates. Morphology, stability, gene expression profiles and chondrogenic potential of micro-aggregates of human and bovine chondrocytes were evaluated and compared to single-cells cultured in micro-wells and in 3D after encapsulation in Dextran-Tyramine (Dex-TA) hydrogels in vitro and in vivo. We successfully formed micro-aggregates of human and bovine chondrocytes with highly controlled size, stability and viability within 24 hours. Micro-aggregates of 100 cells presented a superior balance in Collagen type I and Collagen type II gene expression over single cells and micro-aggregates of 50 and 200 cells. Matrix metalloproteinases 1, 9 and 13 mRNA levels were decreased in micro-aggregates compared to single-cells. Histological and biochemical analysis demonstrated enhanced matrix deposition in constructs seeded with micro-aggregates cultured in vitro and in vivo, compared to single-cell seeded constructs. Whole genome microarray analysis and single gene expression profiles using human chondrocytes confirmed increased expression of cartilage-related genes when chondrocytes were cultured in micro-aggregates. In conclusion, we succeeded in controlled high-throughput formation of micro-aggregates of chondrocytes. Compared to single cell-seeded constructs, seeding of constructs with micro-aggregates greatly improved neo-cartilage formation. Therefore, micro-aggregation prior to chondrocyte implantation in current MACI procedures, may effectively accelerate hyaline cartilage formation

Lytic skull lesion by Leishmaniasis at Makat-Tampu during the Inca Empire: XV-XVI centuries, Rímac Valley, Peru

Paleopathological evidence of mucosal alteration deforming human population of ancient Peru suggests the presence of mucosal leishmaniasis (LM) in the agricultural population living near the endemic area between XV-XVI centuries. The pathological anatomy studies was divided in two phases: 1) pattern of mucosal form in human crania was defined among selected patients, from the Research Center Hospital Evandro Chagas (Fiocruz), with clinical history and destruction of the facial bones, principally oro-nasal cavity. Seven cases were selected. All caused by Leishmania (V.) braziliensis: 6 men and 1 woman with age up to 35 years old. X-ray pictures and axial tomography of the cranium were performed in order to define the pathologic pattern in the bones; 2) archaeological material was analyzed and compared in 241 skulls from the Department of Physical Anthropology of the National Museum of Anthropology, Archaeology and History, Lima, Peru. Material was well preserved and we identified five cases (4 men and 1 woman) with age up to 35 years old presenting naso-palatine destruction, compatible with mucosal form defined by clinical casuistic. It proceeded from the Inca cemetery of Makat-Tampu, Rimac Valley. The rate of mucosal lesions (2.07%) compatible with ML may suggest that there was a high prevalence of L. braziliensis complex infection in the pre-hispanic times and confirm our hypothesis. This study was based on biocultural focus, aiming to reconstruct the quotidian life of the agricultural population of the Rimac valley during the Inca occupation.Evidências paleopatológicas de destruição da mucosa facial em antigas populações humanas do Perú sugerem a presença de Leishmaniose Mucosa (LM) em grupos agrícolas que viveram próximos à área endêmica de LTA entre os séculos XV-XVI. Este estudo foi dividido em duas fases: 1) Através de lâminas de raios X e tomografia axial do crânio, foi definido o padrão de forma mucosa em crânios humanos em sete pacientes (seis homens e uma mulher com mais de 35 anos de idade), selecionados do Centro de Pesquisa Hospital Evandro Chagas (Fiocruz), com história clínica e destruição do maciço facial, principalmente a cavidade oro-nasal. Todos causados por Leishmania (V.) braziliensis', 2) Um material arqueológico bem preservado, constituído de 241 crânios, procedente do cemitério inca de Makat-tampu, vale do Baixo Rímac, foi analisado e comparado. Este se encontra no Departamento de Antropologia Física do Museu Nacional de Antropologia, Arqueologia e História, Lima, Perú. Segundo a observação e descrição anátomopatológica, foram identificados cinco casos (quatro homens e uma mulher acima de 30 anos de idade) com destruição naso-palatina compatíveis com LM. A freqüência desta lesão mucosa (2.07%) pode sugerir que havia uma alta prevalência de infecção pelo complexo L. braziliensis em períodos pré- hispânicos e confirmou nossas hipóteses

Durability assessment of external thermal insulation composite systems in urban and maritime environments

External Thermal Insulation Composite Systems (ETICS) are multilayer solutions which provide an enhanced thermal performance to the building envelope. However, significant anomalies can be detected on ETICS facades, in some cases shortly after the application of these systems. This study intends to evaluate and compare the durability of six commercially available ETICS after two years of outdoor exposure at both urban and maritime conditions in Portugal. The systems were characterized by means of non-destructive testing (i.e., visual and microscopic assessment, water transport properties, thermal conductivity, surface roughness), thus allowing to evaluate the performance loss throughout natural aging. The bio-susceptibility and aesthetic properties (color and gloss) were also investigated. Results showed that the performance and durability of the complete system is significantly affected by the rendering system formulation. The lime-based specimens obtained the highest rate of mold development after one year of aging in a maritime environment, becoming considerably darker and with lower surface gloss. Fungal analysis of this darkish stained area indicated the presence of mold species of the genera Alternaria, Didymella, Cladosporium and Epicoccum, and yeasts of the genera Vishniacozyma and Cystobasidium. An increase of both capillary water absorption and water vapor permeability was also registered for the aged lime-based specimens. Acrylic-based systems obtained lower capillary water absorption after aging and greater dirt deposition on their surfaces, especially in urban conditions. These systems had also higher color variation and surface gloss decrease and slightly higher mold growth, when compared with those aged in a maritime environment. Finally, no mold growth was detected on the silicate-based specimens after two years of aging. However, these specimens obtained higher capillary water absorption and lower vapor permeability after aging, possibly leading to moisture accumulation within the system. Results contribute towards the development of ETICS with enhanced performance and durability.info:eu-repo/semantics/publishedVersio