Darkness of Heart: A Uriel DArcangelo Short Story

The changes in the biochemical compositions and enzymatic activities of rotifer Brachionus plicatilis and Artemia, enriched and stored at 4 degrees C temperature, were determined. The total starvation period was 16 h and samples were taken at the end of the 8th and 16th hours. In present study, the rotifer and nauplii catabolized a large proportion of the protein during the enrichment period. Lipid contents of both live preys increased during the enrichment period and decreased in nauplii and metanauplii throughout the starvation period but lipid content of the rotifer remained relatively constant during the starvation period.

The changes observed in the amino acid compositions of Artemia and the rotifer were statistically significant P rotifer was observed during the enrichment period. A sharp increase in the amounts of docosahexaenoic acid DHA during the enrichment of the rotifer and Artemia nauplii was observed.

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However, the amount of DHA throughout the starvation period decreased in Artemia metanauplii but not in Artemia nauplii. Significant differences in tryptic, leucine aminopeptidase N LAP , and alkaline phosphatase AP enzyme activities of Artemia and rotifer were observed during the enrichment and starvation period P Use of Probiotic Bacillus spp. This study was to evaluate the effect of a preparation of Bacillus probiotic Bacillus licheniformis and B. All treatments were performed in triplicate. Chemical parameters of rearing water viz.

Shrimp larvae were fed Artemia nauplii and rotifers at and individuals per shrimp larvae four times a day, respectively. Lowest total length, weight and survival rate 7. We concluded that Bacillus probiotic can improve growth and survival rate of Pacific white shrimp larvae without conceivably undesirable effects. The reproductive toxicity on the rotifer Brachionus plicatilis induced by BDE and studies on the effective mechanism based on antioxidant defense system changes. The results showed the following: In fact, BDE exerted reproductive inhibition effects in a time- and concentration-dependent manner.

The obtained life tables indicated that BDE at a high concentration prolonged the generation time, whereas low and moderate concentrations of BDE had the opposite effects. BDE at a medium concentration significantly decreased the life expectancy and net reproductive rate P plicatilis. The GST activity was induced significantly by the low concentration of BDE and inhibited by the highest concentration tested. The results indicated that BDE causes reproductive toxicity in B. Intracellular haemolytic agents of Heterocapsa circularisquama exhibit toxic effects on H.

A harmful dinoflagellate, Heterocapsa circularisquama, is highly toxic to shellfish and the zooplankton rotifer Brachionus plicatilis. A previous study found that H. Detailed analysis of the haemolytic activity of H. Interestingly, the ultrasonic-ruptured cells of H. Analysis of samples of the cell contents supernatant and cell fragments precipitate prepared from the ruptured H. Ethanol extract prepared from H.

Analysis on a column of Sephadex LH revealed that light-dependent haemolytic activity and inhibitory activity on cell-mediated light-independent haemolytic activity existed in two separate fractions f-2 and f-3 , suggesting that both. The effects of salinity, pH, and dissolved organic matter on acute copper toxicity to the rotifer , Brachionus plicatilis "L" strain.

This paper presents data from original research for use in the development of a marine biotic ligand model and, ultimately, copper criteria for the protection of estuarine and marine organisms and their uses.

Sacrificed Lives

Ten h static acute unfed copper toxicity tests using the euryhaline rotifer Brachionus plicatilis "L" strain were performed to assess the effects of salinity, pH, and dissolved organic matter measured as dissolved organic carbon; DOC on median lethal dissolved copper concentrations LC Cryptic species are increasingly being recognized in many organisms.

A number of Brachionus cryptic species have been recognized among hatchery strains. In this study, we present a simple, one-step genetic method to detect the presence of those Brachionus sp. With the proposed technique, each of the B. Manjavacas, and Brachionus sp. Based on cytochrome c oxidase subunit I sequences, we reviewed all the available cryptic Brachionus sp. Two highly sensitive protocols were developed for using the primers. Many of the primers can be combined in the same PCR. With this technique, hatchery managers could easily determine their rotifer composition at the level of cryptic species and monitor their cultures more efficiently.

Four marine microalgal species Chlorella autotrophyca, Nannochloropsis gaditana, Tetraiselmis chuii, and Isochrysis aff. After that, a group of 10 neonate rotifers were fed with these four microalgal species. In all cases, cells with preaccumulated metal caused a delay of 1 or 2 days in populational development of rotifers increase in "lag phase". The microalgae that were not fed to rotifers disposed in parallel series did not significantly transfer metal to the medium after the first day. Glycosidases in Brachionus plicatilis Rotifera. Tests for glycosidases were performed in homogenates of Brachionus plicatilis.

Hydrolytic activity was detected with the following substrates: The pH dependence of the glycosidase activities was determined. The distribution of enzyme activities within fractions from the homogenate was studied in order to localize them within the cell.

Proteins from Brachionus homogenate were separated by SDS-gel electrophoresis and the positions of the following glycosidase activities were detected by assays performed on the gels estimated molecular weights in parentheses: Rotifers are an important live feed for first feeding larvae of many fish species.

The use of concentrated algae cells in the mass culture of the rotifer Brachionus plicatilis Brachionidae has opened new horizons for research on this organism. Four different microalgae pastes were prepared: All treatments showed similar contents of proteins and total lipids with respect to control culture a fresh culture of R. The fatty acid profile varied with respect to the control culture, mainly in the proportion of the essential polyunsaturated fatty acids PUFAs: The rotifers grew equally well when fed with centrifuged pastes or control culture maximum density: No significant effect of vitamin C was.

Ingestion of Brachionus plicatilis under different microalgae conditions. The effects of four microalgae, Chlorella vulgaris, Platymonas helgolandicavar, Isochrysis galbana, and Nitzschia closterium on the grazing and filtering rates of the marine rotifer , Brachionus plicatilis , were evaluated under laboratory conditions. The grazing rates in separate cultures of the four microalga were as follows: However, the filtering rates occurred in the following order: A mixed diets experiment revealed that P.

In addition, the grazing rate of B. Furthermore, the filtering rate increased slightly when the density of the microalgae was low, after which it declined as the microalgal density increased. The grazing rates of B. Additionally, the filtering rates during the growth phases were: The results of this study provide foundational information that can be used to explore the optimal culture conditions for rotifers and to promote the development of aquaculture.

Isolation and purification of glutathione S-transferases from Brachionus plicatilis and B. The enzyme glutathione S-transferase GST , a critical element in xenobiotic metabolism, was isolated from the marine rotifer Brachionus plicatilis and its freshwater congener B. Specific activity of crude homogenates was 56 nmol min-1 mg-1 protein, while that of affinity chromatography purified GST was Crude GST specific activity was nmol min-1 mg-1 protein and purified was 72, Rotifer GSTs are unusual because they are monomers whereas all other animals thus far investigated posses dimeric GSTs.

Rapid toxicity assessment using an in vivo enzyme test for Brachionus plicatilis Rotifera. A 1-hr in vivo enzyme inhibition assay based on esterase activity has good potential for marine toxicity assessment.

Search results

A test was developed for the rotifer Brachionus plicatilis based on the nonfluorescent substrate fluorescein diacetate FDA , which is metabolized by esterases to a fluorescent product. Enzyme inhibition, as determined by reduced fluorescence, can be scored visually or quantified using a fluorometer. The 1-hr esterase inhibition test has sensitivity comparable to that of hr rotifer acute tests for several compounds. The toxicity of six compounds was examined using the quantified assay.

The resulting IC20s were within a factor of 3 of the hour LC50s. IC20 values ranged from 0. Electrophoretic analysis of rotifer homogenates suggested that a single C esterase acetylesterase was responsible for FDA metabolism in B. Several other aquatic species are capable of metabolizing FDA, including Brachionus calyciflorus, Mysidopsis bahia, Menidia beryllina, Pimephales promelas, Ceriodaphnia dubia, Daphnia pulex, Artemia salina, and Ophryotrocha sp.

The esterase inhibition test is an attractive tool for assessing aquatic toxicity because of its speed, simplicity, sensitivity, and applicability to a broad range of aquatic species. Measurement of Survival Time in Brachionus Rotifers: Synchronization of Maternal Conditions. Rotifers are microscopic cosmopolitan zooplankton used as models in ecotoxicological and aging studies due to their several advantages such as short lifespan, ease of culture, and parthenogenesis that enables clonal culture. However, caution is required when measuring their survival time as it is affected by maternal age and maternal feeding conditions.

Here we provide a protocol for powerful and reproducible measurement of the survival time in Brachionus rotifers following a careful synchronization of culture conditions over several generations. Empirically, poor synchronization results in early mortality and a gradual decrease in survival rate, thus resulting in weak statistical power. Indeed, under such conditions, calorie restriction CR failed to significantly extend the lifespan of B. This protocol is probably useful for other invertebrate models, including the fruitfly Drosophila melanogaster and the nematode Caenorhabditis elegans, because maternal age effects have also been reported in these species.

The effect of suspended silt from dredging at Yangtze estuary on Brachionus plicatilis was investigated by clonal culture to construct life table and population accumulative culture. The intrinsic increasing rate of the rotifer population was greatly reduced under different concentrations 1, 3, 5, 7 and 9 mg. The suspended silt affected population survival rate, rather than its reproduction rate. In accumulative culture, the densities of female, males, parthenogenetic eggs, and resting eggs in the population were not affected by silt.

It was concluded that the presence of suspended silt should have a certain negative influence on the rotifer population in dredging Yangtze estuary. In a laboratory experiment, Moina mongolica and Brachionus plicatilis were polycultured at four relatively inoculating densities, i. Interspecific interference did exist between M. In the polycultured microcosms, depressed population density of M. Further experiment showed that the mortality of M. The relatively strong capacity of B. Toxic effects of combined effects of anthracene and UV radiation on Brachionus plicatilis.

Anthracene is a typical polycyclic aromatic hydrocarbon, with photo activity, can absorb ultraviolet light a series of chemical reactions, aquatic organisms in the ecosystem has a potential light induced toxicity. In this paper, the effects of anthracene and UV radiation on the light-induced toxicity of Brachionus plicatilis were studied. The main methods and experimental results were as follows: Therefore, the anthracite stress induced by UV radiation could more strongly interfere with the ant oxidative metabolism of B.

Speciation in ancient cryptic species complexes: Continental lake-dwelling zooplanktonic organisms have long been considered cosmopolitan species with little geographic variation in spite of the isolation of their habitats. Evidence of morphological cohesiveness and high dispersal capabilities support this interpretation. However, this view has been challenged recently as many such species have been shown either to comprise cryptic species complexes or to exhibit marked population genetic differentiation and strong phylogeographic structuring at a regional scale.

Here we investigate the molecular phylogeny of the cosmopolitan passively dispersing rotifer Brachionus plicatilis Rotifera: Monogononta species complex using nucleotide sequence variation from both nuclear ribosomal internal transcribed spacer 1, ITS1 and mitochondrial cytochrome c oxidase subunit I, COI genes. Analysis of rotifer resting eggs from 27 salt lakes in the Iberian Peninsula plus lakes from four continents revealed nine genetically divergent lineages.

The high level of sequence divergence, absence of hybridization, and extensive sympatry observed support the specific status of these lineages. Sequence divergence estimates indicate that the B. We discuss these results in relation to the ecology and genetics of aquatic invertebrates possessing dispersive resting propagules and address the apparent contradiction between zooplanktonic population structure and their morphological stasis. Ecological factors affecting gene flow in the Brachionus plicatilis complex Rotifera.

We investigated how adaptation to salinity and temperature acts as reproductive barriers in three sympatric species from the Brachionus plicatilis species complex. These species co-occur in a salt marsh in Spain, and a previous electrophoretic study of variation revealed no hybrids between them. A factorial experiment was designed to test for differences in population growth rates and patterns of bisexual reproduction.

The design combined representative strains from each species in different salinity and temperature conditions, representing the range over which these rotifers are found in their natural environment. We found differences in the growth response of the three species to both factors and in the pattern of bisexual reproduction. These differences help to explain patterns of succession observed in the field. We conclude that these ecological factors, together with mate recognition systems, account for the absence of gene flow in these sympatric species. Persistent genetic signatures of colonization in Brachionus manjavacas rotifers in the Iberian Peninsula.

Recent phylogeographical assessments have consistently shown that continental zooplankton display high levels of population subdivision, despite the high dispersal capacity of their diapausing propagules. As such, there is an apparent paradox between observed cosmopolitanism in the zooplankton that is associated with long-distance dispersal, and strong phylogeographical structures at a regional scale.

Such population dynamics, far from migration-drift equilibrium, have been shown in the rotifer species complex Brachionus plicatilis , a group of over a dozen species inhabiting salt lakes and coastal lagoons worldwide. Here we present the mitochondrial DNA phylogeography of one of these species, Brachionus manjavacas, in the Iberian Peninsula, where it often co-occurs with the morphologically similar species B.

We obtained sequences from individuals from diapausing eggs and clonal cultures from 16 lakes in the Iberian Peninsula, and a Tunisian lake. Phylogenetic and nested clade analysis showed that clade A has a strong phylogeographical structure, with a strong similarity of phylogeographical patterns between B. These include i signatures of allopatric fragmentation between central and southern populations, and ii range expansions in the Iberian Peninsula, both likely to have occurred during the Pleistocene.

We find evidence for a glacial refugium in the Guadiana basin. Clades A and B co-occurred in several of these lakes because of range expansion and secondary contact between both clades. The co-occurrence between B. The strong correlation of genetic and geographical distance found suggests that historical events can lead to such correlation, mirroring the effects of.

In contrast to the highly conserved mitogenomic structure and organisation in most animals including rotifers , the two previously sequenced monogonont rotifer mitogenomes were fragmented into two chromosomes similar in size, each of which possessed one major non-coding region mNCR of about 4—5 Kbp. To further explore this phenomenon, we have sequenced and analysed the mitogenome of one of the most studied monogonont rotifers , Brachionus calyciflorus.

It is also composed of two circular chromosomes, but the chromosome-I is extremely large 27 bp; 3 mNCRs , whereas the chromosome-II is relatively small 9 bp; 1 mNCR. With the total size of 37 bp, it is one of the largest metazoan mitogenomes ever reported. In comparison to other monogononts, gene distribution between the two chromosomes and gene order are different and the number of mNCRs is doubled. Atp8 was not found common in rotifers , and Cytb was present in two copies the first report in rotifers.

A high number 99 of SNPs indicates fast evolution of the Cytb-1 copy. The four mNCRs 5. Publication of this sequence shall contribute to the understanding of the evolutionary history of the unique mitogenomic organisation in this group of rotifers. Postembryonic development of hard jaws trophi in a species belonging to the Brachionus plicatilis complex Rotifera, Monogononta: The presence of hard jaws trophi , with species-specific shape and size, is a taxonomic feature of Rotifera, a group of microscopic metazoans.

Since trophi are used to discriminate among species, it is important to know whether these structures change in taxonomically important ways during postembryonic development. Using both SEM and optical images, we analyzed more than individuals of a single clonal lineage of a monogonont rotifer , Brachionus plicatilis , in order to describe body and trophi development after hatching.

Body size, expressed as lorica width and length, was isometrically related to age of the animals only during preadult stages. Trophi size, expressed as length of the different parts, was unrelated to either age or body size. Therefore, trophi elements do not grow after hatching in B. Despite the dimensional invariance with age, some differences in trophi size among individuals of the same clone were recorded.

No difference in left-right asymmetry of the trophi was shown; the asymmetric elements of the trophi named rami consistently had the right ramus longer than the left. This constancy is in contrast to the reported trophi asymmetries in bdelloid rotifers , in which left-right asymmetries are not constant within clonal lineages. In conclusion, we suggest that also trophi size, constant within the analyzed clone, may be used as an additional taxonomic feature to help in the discrimination of taxa within the B.

Copyright c Wiley-Liss, Inc. Treatment of fish-processing wastewater by co-culture of Candida rugopelliculosa and Brachionus plicatilis. This research was conducted as a part of the continuous development of a novel technique for managing fish-processing wastewater by cultivating proteolytic yeast, Candida rugopelliculosa, as possible diet of the rotifer , Brachionus plicatilis.

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It was feasible to use Alaska Pollack processing wastewater as a growth medium for C. Maximum growth of C. Method of 4th order Runge-Kutta approximation was successfully applied to determine the Monod kinetics of C. The maximum microbial growth rates, mu max , and half saturation coefficient, K s , were determined to be 0. The microbial yield coefficient, Y, and microbial decay rate coefficient, k d , were determined to be 1. Macondo oil mixture represent: Extrapolating these results to the oil released by the Macondo well, suggests underestimation of increased toxicity from Corexit application.

We found small differences in sensitivity among species of the B. Long-term survival of hydrated resting eggs from Brachionus plicatilis. Several organisms display dormancy and developmental arrest at embryonic stages. Long-term survival in the dormant form is usually associated with desiccation, orthodox plant seeds and Artemia cysts being well documented examples.

Several aquatic invertebrates display dormancy during embryonic development and survive for tens or even hundreds of years in a hydrated form, raising the question of whether survival in the non-desiccated form of embryonic development depends on pathways similar to those occurring in desiccation tolerant forms. To address this question, Illumina short read sequencing was used to generate transcription profiles from the resting and amictic eggs of an aquatic invertebrate, the rotifer , Brachionus plicatilis.

These two types of egg have very different life histories, with the dormant or diapausing resting eggs, the result of the sexual cycle and amictic eggs, the non-dormant products of the asexual cycle. Significant transcriptional differences were found between the two types of egg, with amictic eggs rich in genes involved in the morphological development into a juvenile rotifer. In contrast, representatives of classical "stress" proteins: More importantly however, was the identification of transcripts for messenger ribonucleoprotein particles which stabilise RNA.

These inhibit translation and provide a valuable source of useful RNAs which can be rapidly activated on the exit from dormancy. Apoptotic genes were also present. Although apoptosis is inconsistent with maintenance of prolonged dormancy, an altered apoptotic pathway has been proposed for Artemia, and this may be the case with the rotifer. These data represent the first transcriptional profiling of molecular processes associated with dormancy in a non-desiccated form and indicate important similarities in the molecular pathways activated in resting eggs compared.

Background Several organisms display dormancy and developmental arrest at embryonic stages. Conclusions These data represent the first transcriptional profiling of molecular processes associated with dormancy in a non-desiccated form and indicate important similarities in. Effect of the nutritional status of semi-continuous microalgal cultures on the productivity and biochemical composition of Brachionus plicatilis. The rotifer Brachionus plicatilis was cultured using the microalga Isochrysis aff. The increase of renewal rate led to increasing nutrient and light availability in microalgal cultures, which caused differences in the biochemical composition of microalgal biomass.

Growth rate, individual dry weight, organic content, and biomass productivity of rotifer cultures increased in response to higher growth rate in T-ISO cultures. Consequently, biomass productivity of rotifer cultures increased more than twofold with the increase of renewal rate of T-ISO cultures. Rotifer organic content underwent the same trend of total dry weight. Selecting the most appropriate conditions for microalgal culture can therefore enhance the nutritive quality of microalgal biomass, resulting in a better performance of filter feeders and their nutrient content, and may constitute a useful tool to improve the rearing of fish larvae and other aquaculture organisms that require live feed in some or all the stages of their life cycle.

Acute toxicity testing method was used to study effects of different density of Prorocentrum donghaiense, Heterosigma akashiwo and Alexandrium tamarense on mortality rates and population growth parameter of Brachionus plicatilis under controlled experimental conditions. Results showed that 24 h LC50 values of Prorocentrum donghaiense, Heterosigma akashiwo and Alexandrium tamarense treatment to mortality rate of Brachionus plicatilis were 3. Marked density effects were presented when three species of red tide microalga showed their toxicity to Brachionus plicatilis.

Inhibitory effects of three species of microalga on Brachionus plicatilis were enhanced with increasing of microalgal density. While marine invertebrate cytochrome P CYP genes and their roles in detoxification mechanisms have been studied, little information is available regarding freshwater rotifer CYPs and their functions. The 31 Bc-CYP genes with a few tandem duplications were clustered into CYP 2, 3, 4, mitochondrial, and 46 clans with two marine rotifers Brachionus plicatilis and Brachionus koreanus. Complete mitochondrial genome of the monogonont rotifer , Brachionus koreanus Rotifera, Brachionidae.

The complete mitochondrial genome was obtained from the assembled genome data sequenced by next generation sequencing NGS technology from the monogonont rotifer Brachionus koreanus. The mitochondrial genome of B. The gene contents of B.

However, gene orders of B. They also showed anti-G bias Influence of temperature and conductivity on the life-history characteristics of a pampean strain of Brachionus plicatilis. In the present work, we provide the first approach about the life-history of Brachionus plicatilis in South America. We tested with laboratory experiments the response of the pampean strain of B. We evaluated the effects of eight conductivity values from 1 to 17 mS.

The results demonstrated that the increase in population-growth rate and the peak of abundance occurred at the highest temperature and at medium conductivity. Minimum values were obtained at the lowest temperature and conductivities analyzed, but the final density attained was nevertheless similar to those recorded in the pampean environments at the optimum conductivity and during the spring and summer seasons. Males, mictic females, and resting eggs were observed at the minimum and maximum conductivities, revealing the strategy of this species for maintaining dominance in environments with fluctuating salinity.

The experiments also indicated the possible behavior of this relevant member of the zooplankton community within a scenario of increasing temperature and salinity related to the climate changes occurring in the pampean region. Genome size evolution at the speciation level: Studies on genome size variation in animals are rarely done at lower taxonomic levels, e. Yet, such variation might provide important clues on the tempo and mode of genome size evolution. In this study we used the flow-cytometry method to study the evolution of genome size in the rotifer Brachionus plicatilis , a cryptic species complex consisting of at least 14 closely related species.

We found an unexpectedly high variation in this species complex, with genome sizes ranging approximately seven-fold haploid '1C' genome sizes: In one genealogical species, called B. Overall, genome size was significantly correlated to egg size and body size, even though the latter became non-significant after controlling for phylogenetic non-independence. Our study suggests that substantial genome size variation can build up early during speciation, potentially even among isolated populations.

An alternative, but not mutually exclusive interpretation might be that reproductive isolation tends to build up unusually slow in this species complex. The cryptic species complex Brachionus plicatilis Rotifera. Background Studies on genome size variation in animals are rarely done at lower taxonomic levels, e.

Results We found an unexpectedly high variation in this species complex, with genome sizes ranging approximately seven-fold haploid '1C' genome sizes: Conclusions Our study suggests that substantial genome size variation can build up early during speciation, potentially even among isolated populations. Effect of periodical starvation on the life history of Brachionus plicatilis O.

To estimate the changes in the life history of the rotifer Brachionus plicatilis O. Rotifers were fed for h daily, and were then starved until the next day. Control animals were fed throughout their lifespan. Starved rotifers matured and produced their first offspring at an older age than the control animals.

The periodical starvation resulted in a decrease in the lifetime fecundity to less than half that of the non-starved control. The reproductive period and lifespan were times longer in the starved animals than in the control animals. The negative relationship between lifespan and lifetime fecundity is interpreted as a trade-off in an alternative life-history strategy of rotifers under starved conditions. The great decrease in fecundity and extension of lifespan enables rotifers to compensate to keep the population in equilibrium.

Swimming speed alteration of Artemia sp. In this study, we investigated the possibility to improve a new behavioural bioassay Swimming Speed Alteration test-SSA test using larvae of marine cyst-forming organisms: Swimming speed was investigated as a behavioural end-point for application in ecotoxicology studies. A first experiment to analyse the linear swimming speed of the two organisms was performed to verify the applicability of the video-camera tracking system, here referred to as Swimming Behavioural Recorder SBR.

A second experiment was performed, exposing organisms to different toxic compounds zinc pyrithione, Macrotrol MT, and Eserine. Swimming speed alteration was analyzed together with mortality. The results of the first experiment indicate that SBR is a suitable tool to detect linear swimming speed of the two organisms, since the values have been obtained in accordance with other studies using the same organisms 3. Toxicity test results clearly indicate that swimming speed of Artemia sp. Indeed, alterations in swimming speed have been detected at toxic compound concentrations as low as less then 0.

In conclusion, the SSA test with B. The mate recognition protein gene mediates reproductive isolation and speciation in the Brachionus plicatilis cryptic species complex. Background Chemically mediated prezygotic barriers to reproduction likely play an important role in speciation. In facultatively sexual monogonont rotifers from the Brachionus plicatilis cryptic species complex, mate recognition of females by males is mediated by the Mate Recognition Protein MRP , a globular glycoprotein on the surface of females, encoded by the mmr-b gene family.

In this study, we sequenced mmr-b copies from 27 isolates representing 11 phylotypes of the B. Results Isolates of the B. The repeats within a gene copy are generally more similar than are gene copies among phylotypes, suggesting concerted evolution. Compared to housekeeping genes from the same isolates, mmr-b has accumulated only half as many synonymous differences but twice as many non-synonymous differences.

Most of the amino acid differences between repeats appear to occur on the outer face of the protein, and these often result in changes in predicted patterns of phosphorylation. However, we found no evidence of positive selection driving these differences. Isolates with the most divergent copies were unable to mate with other isolates and rarely self-crossed.

Overall the degree of mate recognition was significantly correlated with the genetic distance of mmr-b. Conclusions Discrimination of compatible mates in the B. Microstructured implant surfaces created by grit blasting and acid etching titanium Ti support osseointegration. This effect is further enhanced by storing in aqueous solution to retain hydrophilicity, but this also leads to surface nanostructure formation.

The purpose of this study was to assess the contributions of nanostructures on the improved osteogenic response of osteoblast lineage cells to hydrophilic microstructured Ti. Contact angle measurements indicated only modSLA was hydrophilic.

Confocal laser microscopy revealed minor differences in mean surface roughness. These results suggest that when the appropriate microstructure is present, hydrophilicity may play a greater role in stimulating MSC and NHOst osteoblastic differentiation and maturation than the presence of nanostructures generated during storage in an aqueous environment. Generation and characterization of two immortalized human osteoblastic cell lines useful for epigenetic studies.

Different model systems using osteoblastic cell lines have been developed to help understand the process of bone formation. Here, we report the establishment of two human osteoblastic cell lines obtained from primary cultures upon transduction of immortalizing genes. The resulting cell lines had no major differences to their parental lines in their gene expression profiles. Similar to primary osteoblastic cells, osteocalcin transcription increased following 1,dihydroxyvitamin D 3 treatment and the immortalized cells formed a mineralized matrix, as detected by Alizarin Red staining.

Moreover, these human cell lines responded by upregulating ALPL gene expression after treatment with the demethylating agent 5-aza-2'-deoxycytidine AzadC , as shown before for primary osteoblasts. More importantly, we show that these cells respond to demethylating treatment, as shown by the increase in SOST mRNA levels, the gene encoding sclerostin, upon treatment of the recipient mice with AzadC.

Altogether our results show that these immortalized cell lines constitute a particularly useful model system to obtain further insight into bone homeostasis, and particularly into the epigenetic mechanisms regulating sclerostin production. Osteoinductive effect of bone bank allografts on human osteoblasts in culture. Incorporation of a human bone allograft requires osteoclast activity and growth of recipient osteoblasts. The aim of this work was to study the effects produced by autoclavated and degrees C frozen bone allografts on osteoblast proliferation and synthesis of interleukin 6 IL6 , activator of bone resorption, aminoterminal propeptide of procollagen I PINP , marker of bone matrix formation, and osteoprotegerin OPG , inhibitor of osteoclast activity and differentiation.

Allografts were obtained from human femoral heads. Human osteoblasts were cultured in the presence problem group or in the absence control group of allografts during 15 days. Allografts produced a decrease in osteoblast proliferation in the first week of the experiment, and an increase in IL6 mRNA, both at 3 h and 2 days, and an increase in the IL6 released to the culture medium the second day of the experiment. We found a decrease in OPG released to the culture on the 2nd and fourth days. These results suggest an increase in bone resorption and a decrease in bone formation in the first week of the experiment.

In the second week, allografts produced an increase in osteoblast proliferation and PINP release to the culture medium, indicating an increase in bone formation; an increase in OPG released to the culture medium, which would indicate a decrease in bone resorption; and a decrease in IL6, indicating a decrease in bone resorption stimulation.

These results demonstrate that autoclavated and degrees C frozen bone allografts produce in bone environment changes that regulate their own incorporation to the recipient bone. Zirconium Zr is an element commonly used in dental and orthopedic implants either as zirconia ZrO2 or in metal alloys. It can also be incorporated into calcium silicate-based ceramics.

However, the effects of in vitro culture of human osteoblasts HOBs with soluble ionic forms of Zr have not been determined. Incubation of human osteoblast cultures with Zr ions increased the proliferation of human osteoblasts and also gene expression of genetic markers of osteoblast differentiation. In conclusion, Zr ions appear able to induce both the proliferation and the differentiation of primary human osteoblasts. Expression of extracellular calcium-sensing receptor in human osteoblastic MG cell line. The existence of the CaR in osteoblasts remains controversial, however, since some studies have failed to document its expression in the same osteoblast -like cell lines.

The goals of the present study were twofold. Polyelectrolyte multilayer PEMU coatings built layer by layer with alternating pairs of polyelectrolytes can be tuned to improve cell interactions with surfaces and may be useful as biocompatible coatings to improve fixation between implants and tissues. Characterization of cadmium uptake and cytotoxicity in human osteoblast -like MG cells. Since bone mass is maintained constant by the balance between osteoclastic bone resorption and osteoblastic bone formation, alterations in osteoblast proliferation and differentiation may disturb the equilibrium of bone remodeling.

Exposure to cadmium Cd has been associated with the alteration of bone metabolism and the development of osteoporosis. Because little information is available about the direct effects of Cd on osteoblastic cells, we have characterized in vitro the cellular accumulation and cytotoxicity of Cd in human osteoblastic cells. In order to characterize the mechanisms of Cd uptake, experiments have been conducted under well-defined metal speciation conditions in chloride and nitrate transport media. The cellular accumulation and cytotoxicity of Cd increased in the absence of extracellular calcium Ca , suggesting that Cd may enter the cells in part through Ca channels.

However, neither the cellular accumulation nor the cytotoxicity of Cd was modified by voltage-dependent Ca channel VDCC modulators or potassium-induced depolarization. Moreover, exposure conditions activating or inhibiting capacitative Ca entry CCE failed to modify the cellular accumulation and cytotoxicity of Cd, which excludes the involvement of canonical transient receptor potential TRPC channels. The inhibition of Cd uptake by Mg and Ca was not additive.

The cells were identified by cell morphology, calcium nodule formation and alkaline phosphatase ALP activity assay. The osteoblasts obtained were positive for ALP staining and could form calcium nodules in vitro. The drugs also increased the relative proliferation index of the osteoblasts , and high concentration of the drugs resulted in greater number of the mineral nodules in the osteoblasts P osteoblasts in vitro.

Detachment strength of human osteoblasts cultured on hydroxyapatite with various surface roughness. Contribution of integrin subunits. Hydroxyapatite HA has been widely used as a bone substitute in dental, maxillofacial and orthopaedic surgery and as osteoconductive bone substitute or precoating of pedicle screws and cages in spine surgery. The aim of the present study was to investigate the osteoblastic adhesion strength on HA substrata with different surface topography and biochemistry pre-adsorption of fibronectin after blocking of specific integrin subunits with monoclonal antibodies.

Human bone marrow derived osteoblasts were cultured on HA disc-shaped substrata which were sintered and polished resulting in two surface roughness grades. For attachment evaluation, cells were incubated with monoclonal antibodies and seeded for 2 h on the substrata. Cell detachment strength was determined using a rotating disc device.

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Cell detachment strength was surface roughness, fibronectin preadsorption and intergin subunit sensitive. Adenosine Triphosphate stimulates differentiation and mineralization in human osteoblast -like Saos-2 cells. In the last years adenosine triphosphate ATP and subsequent purinergic system activation through P2 receptors were investigated highlighting their pivotal role in bone tissue biology. In osteoblasts ATP can regulate several activities like cell proliferation, cell death, cell differentiation and matrix mineralization.

Since controversial results exist, in this study we analyzed the ATP effects on differentiation and mineralization in human osteoblast -like Saos-2 cells. We showed for the first time the altered functional activity of ATP receptors. Moreover, we found that ATP can affect mineralization in a biphasic manner, at low concentrations ATP always increases mineral deposition while at high concentrations it always reduces mineral deposition.

In conclusion, we show the osteogenic effect of ATP on both early and late stage activities like differentiation and mineralization, for the first time in human osteoblastic cells. Apoptosis may determine the release of skeletal alkaline phosphatase activity from human osteoblast -line cells. Although quantitative measurement of skeletal alkaline phosphatase sALP activity in serum can provide an index of the rate of bone formation, the metabolic process that determines the release of sALP - from the surface of osteoblasts , into circulation-is unknown. The current studies were intended to examine the hypothesis that the release of sALP from human osteoblasts is a consequence of apoptotic cell death.

We measured the release of sALP activity from human osteosarcoma SaOS-2 cells and normal human bone cells, under basal conditions and in response to agents that increased apoptosis TNF-a, okadiac acid and agents that inhibit apoptosis IGF-I, calpain, and caspase inhibitors. Apoptosis was determined by the presence of nucleosomes histone-associated DNA in the cytoplasm of the cells by using a commercial kit. Together, these data demonstrate that the.

Human osteoblast cells were isolated from bone tissue and characterized by immunofluorescence Staining before and after purification using magnetic bead system. The results have shown that the human osteoblast cells were successfully purified and were suitable for subsequent cell culturing process. Expression of bone-specific genes was taken place after the cells were incubated in composite extract solutions. Furthermore, osteoblast cells were able to mineralize the matrix next to composite samples.

Scanning electron microscopy demonstrated that cells had normal morphology on the scaffold. Patients with HIV infection have decreased numbers of osteoblasts , decreased bone mineral density and increased risk of fracture compared to uninfected patients; however, the molecular mechanisms behind these associations remain unclear. We questioned whether Gp, a component of the envelope protein of HIV capable of inducing apoptosis in many cell types, is able to induce cell death in bone-forming osteoblasts. We show that treatment of immortalized osteoblast -like cells and primary human osteoblasts with exogenous Gp in vitro at physiologic concentrations does not result in apoptosis.

Instead, in the osteoblast -like U2OS cell line, cells expressing CXCR4, a receptor for Gp, had increased proliferation when treated with Gp compared to control P human osteoblasts and likely does not directly contribute to osteoporosis in infected patients by this mechanism. Key role of the expression of bone morphogenetic proteins in increasing the osteogenic activity of osteoblast -like cells exposed to shock waves and seeded on bioactive glass-ceramic scaffolds for bone tissue engineering.

In this work, the role of shock wave-induced increase of bone morphogenetic proteins in modulating the osteogenic properties of osteoblast -like cells seeded on a bioactive scaffold was investigated using gremlin as a bone morphogenetic protein antagonist. Bone-like glass-ceramic scaffolds, based on a silicate experimental bioactive glass developed at the Politecnico di Torino, were produced by the sponge replication method and used as porous substrates for cell culture. Human MG cells, exposed to shock waves and seeded on the scaffolds, were treated with gremlin every two days and analysed after 20 days for the expression of osteoblast differentiation markers.

Shock waves have been shown to induce osteogenic activity mediated by increased expression of alkaline phosphatase, osteocalcin, type I collagen, BMP-4 and BMP Cells exposed to shock waves plus gremlin showed increased growth in comparison with cells treated with shock waves alone and, conversely, mRNA contents of alkaline phosphatase and osteocalcin were significantly lower. Therefore, the shock wave-mediated increased expression of bone morphogenetic protein in MG cells seeded on the scaffolds is essential in improving osteogenic activity; blocking bone morphogenetic protein via gremlin completely prevents the increase of alkaline phosphatase and osteocalcin.

The results confirmed that the combination of glass-ceramic scaffolds and shock waves exposure could be used to significantly improve osteogenesis opening new perspectives for bone regenerative medicine. Therapeutic touch affects DNA synthesis and mineralization of human osteoblasts in culture. Complementary and alternative medicine CAM techniques are commonly used in hospitals and private medical facilities; however, the effectiveness of many of these practices has not been thoroughly studied in a scientific manner.

Dolores Krieger and Dora Kunz, Therapeutic Touch is one of these CAM practices and is a highly disciplined five-step process by which a practitioner can generate energy through their hands to promote healing. There are numerous clinical studies on the effects of TT but few in vitro studies. Our purpose was to determine if Therapeutic Touch had any effect on osteoblast proliferation, differentiation, and mineralization in vitro.

TT was performed twice a week for 10 min each on human osteoblasts HOBs and on an osteosarcoma-derived cell line, SaOs In conclusion, Therapeutic Touch appears to increase human osteoblast DNA synthesis, differentiation and mineralization, and decrease differentiation and mineralization in a human osteosarcoma-derived cell line. Published by Wiley Periodicals, Inc. Development of biomimetic nanocomposites as bone extracellular matrix for human osteoblastic cells. Here, we have developed biomimetic nanocomposites containing chitosan, poly vinyl alcohol and nano-hydroxyapatite-zinc oxide as bone extracellular matrix for human osteoblastic cells and characterized by Fourier transform infrared spectroscopy, powder X-ray diffraction.

Scanning electron microscopy images revealed interconnected macroporous structures. These values are similar to human cancellous bone. These nanocomposites also showed superior water uptake, antimicrobial and biodegradable properties than the previously reported results. Compatibility with human blood and pH was observed, indicating nontoxicity of these materials to the human body. Moreover, proliferation of osteoblastic MG cells onto the nanocomposites was also observed without having any negative effect.

Monolayer formation of human osteoblastic cells on vertically aligned multiwalled carbon nanotube scaffolds. Monolayer formation of SaOS-2 human osteoblast -like cells was observed on VACNT vertically aligned multiwalled carbon nanotubes scaffolds without purification or functionalization. The VACNT were produced by a microwave plasma chemical vapour deposition on titanium surfaces with nickel or iron as catalyst. Cell viability and morphology studies were evaluated by LDH lactate dehydrogenase release assay and SEM scanning electron microscopy , respectively.

The non-toxicity and the flat spreading with monolayer formation of the SaOs-2 on VACNT scaffolds surface indicate that they can be used for biomedical applications. Tadalafil modulates aromatase activity and androgen receptor expression in a human osteoblastic cell in vitro model. We aimed to characterize, in a cellular model system in vitro, the potential modulation of aromatase and sex steroid hormone receptors upon exposure to tadalafil TAD. Human osteoblast -like cells SAOS-2 were chosen as an in vitro model system since osteoblasts are target of steroid hormones.

Cells were tested for viability upon TAD exposure, which increased cell proliferation. These findings might represent the first evidence of translational actions of PDE5i on AR, which leads to hypothesize a growing relevance of this molecule in men with prostate cancer long-term treated with TAD for sexual rehabilitation. Recently, it was shown that LF also regulates bone cell activity, suggesting its therapeutic effect on postmenopausal bone loss.

However, a minimal amount is known regarding the effects of recombinant human LF rhLF supplementation on bone status in young healthy infants. We found osteoblast cell differentiation was significantly promoted in vitro. In order to investigate the effects of rhLF on bone status in vivo, we used a piglet model, which is a useful model for human infants. Piglets were supplemented with rhLF milk for 30 days.

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Bone formation markers, Serum calcium concentration, bone mineral density BMD , bone mineral content BMC , tibia bone strength, and the overall metabolite profile analysis showed that rhLF was advantageous to the bone growth in piglets. These findings suggest that rhLF supplementation benefits neonate bone health by modulating bone formation.

Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering. The nature of the extracellular matrix ECM is crucial in regulating cell functions via cell-matrix interactions, cytoskeletal organization, and integrin-mediated signaling. For bone tissue engineering, the ECM should also be considered in terms of its function in mediating cell adhesion to biomaterials. This study examined ECM production, cytoskeletal organization, and adhesion of primary human osteoblastic cells on biodegradable matrices applicable for tissue engineering, namely polylactic-co-glycolic acid We hypothesized that the osteocompatible, biodegradable polymer surfaces promote the production of bone-specific ECM proteins in a manner dependent on polymer composition.

We first examined whether the PLAGA and PLA matrices could support human osteoblastic cell growth by measuring cell adhesion at 3, 6 and 12h post-plating. Immunofluorescence revealed an extracellular distribution for CO and FN, whereas OP and ON were found both intracellularly as well as extracellularly on the polymer. In summary, we found that osteoblasts plated on PLAGA adhered better to the substrate, produced higher levels of ECM molecules, and showed greater cytoskeletal. The biomaterials used to maintain or replace functions in the human body consist mainly of metals, ceramics or polymers.

In orthopedic surgery, metallic materials, especially titanium and its alloys, are the most common, due to their excellent mechanical properties, corrosion resistance, and biocompatibility. Aside from the established Ti6Al4V alloy, shape memory materials such as nickel-titanium NiTi have risen in importance, but are also discussed because of the adverse effects of nickel ions.

These might be reduced by specific surface modifications. In the present in vitro study, the osteoblastic cell line MG as well as primary human osteoblasts , fibroblasts, and macrophages were cultured on titanium alloys forged Ti6Al4V, additive manufactured Ti6Al4V, NiTi, and Diamond-Like-Carbon DLC -coated NiTi to verify their specific biocompatibility and inflammatory potential.

DLC-coated NiTi appeared as a suitable surface for cell growth, showing the highest collagen production. None of the implant materials caused a strong inflammatory response. In general, no distinct cell-specific response could be observed for the materials and surface coating used. In summary, all tested titanium alloys seem to be biologically appropriate for application in orthopedic surgery.

Data supporting attempted caveolae-mediated phagocytosis of surface-fixed micro-pillars by human osteoblasts. Caveolin-1 Cav-1 as major structural component of caveolae [2] plays an important role in the phagocytic process of micro-particles and -pillars. Data related to the experiments in [1] with siRNA-mediated knockdown are presented here as well as micro-particle control experiments, tubulin analysis on the micro-pillared topography and initial cell interaction with the micro-pillars. Rooibos flavonoids, orientin and luteolin, stimulate mineralization in human osteoblasts through the Wnt pathway.

Several epidemiological studies have shown that tea consumption is associated with higher bone mineral density in women. Flavonoids in tea are recognized as potential estrogen mimics and may positively influence bone metabolism in estrogen-deficient women. Luteolin and orientin, flavonoids from rooibos tea, are of particular interest as rooibos tea contains no caffeine that can be detrimental to bone health. This study analyzed changes in mineral content when luteolin or orientin was added to a human osteoblast cell line and the potential mechanisms involved. Measurements included alkaline phosphatase ALP activity, cell mitochondrial activity, toxicity, and changes in regulatory proteins involved in osteoblast metabolism.

Mineral was significantly elevated in Saos2 cells treated with orientin 0. Orientin and luteolin, flavonoids in rooibos tea, enhance mineral content in Saos2 cells. These findings provide guidance for doses to be studied in well-established animal models. Proteinase-activated receptor PAR -2 activation impacts bone resorptive properties of human osteoarthritic subchondral bone osteoblasts.

In osteoarthritis OA , the subchondral bone undergoes a remodelling process involving several factors synthesized by osteoblasts. In this study, we investigated the expression, production, modulation, and role of PAR-2 in human OA subchondral bone osteoblasts. PAR-2 expression and production p human OA subchondral bone osteoblasts were compared to normal.

Interestingly, bone resorptive activity was also significantly enhanced following PAR-2 activation. This study is the first to demonstrate that PAR-2 activation plays a role in OA subchondral bone resorption via an up-regulation of major bone remodelling factors.

Proliferation and osteoblastic differentiation of human bone marrow-derived stromal cells on akermanite-bioactive ceramics. In the present study, the effects of a calcium magnesium silicate bioactive ceramic akermanite on proliferation and osteoblastic differentiation of human bone marrow stromal cells hBMSC have been investigated and compared with the classical ceramic beta-tricalcium phosphate, beta-TCP. Proliferation and osteoblastic differentiation were evaluated on day 1, 4, 7 and Furthermore, this stimulatory effect was observed not only in osteogenic medium, but also in normal growth medium without osteogenic reagents such as l-ascorbic acid, glycerophosphate and dexamethasone.

This result suggests that akermanite can promote osteoblastic differentiation of hBMSC in vitro even without osteogenic reagents, and may be used as a bioactive material for bone regeneration and tissue engineering applications. Suppression of osteoblastic phenotypes and modulation of pro- and anti-apoptotic features in normal human osteoblastic cells under a vector-averaged gravity condition. Spaceflight and bed rest induce loss of bone mass.

A number of in vivo and in vitro studies have been conducted to clarify the mechanisms, however, the results have been conflicting. The purpose of this study was to investigate the effects of gravity unloading on proliferation, phenotypes, and apoptosis of normal human osteoblastic cells in the presence of 1alpha,dihydroxyvitamin D3. We used a vector-averaged gravity condition generated by clinostat rotation to simulate gravity unloading. Clinostat rotation did not affect the cell proliferation.

Reduction of osteoblast responsiveness to 1alpha,dihydroxyvitamin D3 might be involved in osteopenia that is induced by gravity unloading. Biphasic positive effect of formononetin on metabolic activity of human normal and osteoarthritic subchondral osteoblasts. Osteoarthritis is a multifactorial disease characterized by loss of articular cartilage and subchondral plate thickening.

Therefore, biochemical analysis of the underlying bone tissue has provided important information for treatment of osteoarthritis. In this study, we determined the potential role of formononetin, a phytoestrogen isolated from Astragalus membranaceus to alter the expression of metabolic markers and cytokine production of human normal osteoblasts Obs and osteoarthritis subchondral osteoblasts OA Obs. Human OA Obs and normal Obs were cultured for 3days, 7days or 14days in the present medium only or were treated with various doses of formononetin.

The remodeling effect of formononetin on osteogenic markers and cytokines of inflammatory mediators was more striking in OA Obs as well. Taken together, these results could suggest that formononetin has biphasic positive effects on normal Obs and OA Obs by modifying their biological synthetic capacities. Published by Elsevier B.

Magnesium alloys have been identified as a new generation material of orthopaedic implants. For the first time, the direct, active, influence of human primary osteoblasts on magnesium-based materials pure magnesium, Mg-2Ag and MgGd alloys is studied for up to 14 days. Several parameters such as composition of the degradation interface directly beneath the cells are analysed with a scanning electron microscope equipped with energy dispersive X-ray and focused ion beam.

Furthermore, influence of the materials on cell metabolism is examined via different parameters like active mineralisation process. The results are highlighting the influences of the selected alloying element on the initial cells metabolic activity. Diabetes mellitus DM is a major risk factor for periodontal disease and affects various cellular functions. The aim of this study is to investigate whether hyperglycemia affects periodontal tissue regeneration, using human PDLSCs and high-glucose medium as a model of DM.

PDLSCs were obtained from healthy adult human mandibular third molars. Cell proliferation, osteoblastic differentiation, and proinflammatory cytokine expression were investigated by culturing PDLSCs in media supplemented with four different glucose concentrations representative of control patients 5.

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The results of this study demonstrate that hyperglycemia inhibits human PDLSC proliferation and osteoblastic differentiation. Transduction of Oct6 or Oct9 gene concomitant with Myc family gene induced osteoblast -like phenotypic conversion in normal human fibroblasts. Osteoblasts play essential roles in bone formation and regeneration, while they have low proliferation potential. Recently we established a procedure to directly convert human fibroblasts into osteoblasts dOBs. The Oct9 plus N-myc was the best combination to induce direct conversion of human fibroblasts into osteoblast -like cells.

The present findings may greatly contribute to the elucidation of the roles of the Oct and Myc proteins in osteoblast direct reprogramming. The results may also lead to establishment of novel regenerative therapy for various bone resorption diseases. Inhibition of fatty acid biosynthesis prevents adipocyte lipotoxicity on human osteoblasts in vitro. Abstract Although increased bone marrow fat in age-related bone loss has been associated with lower trabecular mass, the underlying mechanism responsible remains unknown.

We hypothesized that marrow adipocytes exert a lipotoxic effect on osteoblast function and survival through the reversible biosynthesis of fatty acids FA into the bone marrow microenvironment. We have used a two-chamber system to co-culture normal human osteoblasts NHOst with differentiating pre-adipocytes in the absence or presence of an inhibitor of FA synthase cerulenin and separated by an insert that allowed unidirectional trafficking of soluble factors only and prevented direct cell—cell contact.

Supernatants were assayed for the presence of FA using mass spectophotometry. After 3 weeks in co-culture, NHOst showed significantly lower levels of differentiation and function based on lower mineralization and expression of alkaline phosphatase, osterix, osteocalcin and Runx2. These toxic effects were inhibited by addition of cerulenin.

Furthermore, culture of NHOst with either adipocyte-conditioned media alone in the absence of adipocytes themselves or with the addition of the most predominant FA stearate or palmitate produced similar toxic results. Finally, Runx2 nuclear binding was affected by addition of either adipocyte conditioned media or FA into the osteogenic media. We conclude that the presence of FA within the marrow milieu can contribute to the age-related changes in bone mass and can be prevented by the inhibition of FA synthase. Ectopic bone formation in nude rats using human osteoblasts seeded poly 3 hydroxybutyrate embroidery and hydroxyapatite-collagen tapes constructs.

The aim of this study was to evaluate the ectopic bone formation using tissue engineered cell-seeded constructs with two different scaffolds and primary human maxillary osteoblasts in nude rats over an implantation period of up to 96 days. A total of 72 implants were placed into the back muscles of 18 rats. The osteoblastic character of the cells was confirmed by immunocytochemistry and RT-PCR for osteocalcin. Histological analysis demonstrated that all cell-seeded constructs induced ectopic bone formation after 24, 48 and 96 days of implantation. There was more mineralized tissue in PHB constructs than in HAP-collagen tapes at day 24; p osteoblastic character of the cell-seeded constructs after implantation time.

No bone formation and no osteocalcin expression were found in the control groups. However, the amount of bone formed decreased with increasing length of implantation. Interleukin-7 is a critical cytokine for lymphoid development and a direct inhibitor of in vitro osteoclastogenesis in murine bone marrow cultures.

To explore the role of IL-7 in bone, we generated transgenic mouse lines bearing the 2. In addition we crossed these mice with IL-7 deficient mice to determine if the alterations in lymphopoiesis, bone mass and osteoclast formation observed in the IL-7 KO mice could be rescued by osteoblast -specific overexpression of IL Furthermore, targeted overexpression of IL-7 in osteoblasts rescued the osteopenic bone phenotype and B cell development of IL-7 KO mice but did not have an effect on T lymphopoiesis, which occurs in the periphery.

These results likely reflect both a direct inhibitory effects of IL-7 on osteoclastogenesis in vivo and gender specific differences in responses to IL Eluted zinc ions stimulate osteoblast differentiation and mineralization in human dental pulp stem cells for bone tissue engineering.

Zinc is an essential element for proliferation, differentiation and survival in various cell types. In a previous study, we found that zinc ions released from zinc-modified titanium surfaces eluted zinc ions; EZ stimulate cell viability, osteoblast marker gene expression and calcium deposition in human bone marrow-derived mesenchymal cells hBMCs. The aim of the present study was to investigate the effects of EZ on osteoblast differentiation among dental pulp stem cells DPSCs in vitro. Osteoblast and osteoclast marker gene expression was evaluated by real-time PCR.

We also evaluated alkaline phosphatase ALP staining and calcium deposition. Collectively, these results demonstrate the potential for clinical application to prospective treatment of bone diseases. It is well established that bone forming cells osteoblasts secrete proteins with autocrine, paracrine, and endocrine function. However, the identity and functional role for the majority of these secreted and differentially expressed proteins during the osteoblast OB differentiation process, is not fully established.

To address these questions, we quantified the temporal dynamics of the human stromal mesenchymal, skeletal stem cell hMSC secretome during ex vivo OB differentiation using stable isotope labeling by amino acids in cell culture SILAC. In addition, we employed pulsed SILAC labeling to distinguish genuine secreted proteins from intracellular contaminants.

We identified potentially secreted proteins that were quantified at 5 time-points during days ex vivo OB differentiation including 41 proteins known to be involved in OB functions. Among these, proteins exhibited more than 2-fold up or down-regulation. The pulsed SILAC method revealed a strong correlation between the fraction of isotope labeling and the subset of proteins known to be secreted and involved in OB differentiation.

Furthermore, we studied the biological effects of one of these proteins, the hormone stanniocalcin 2 STC2 and demonstrated its autocrine effects in enhancing osteoblastic differentiation of hMSC. Our study demonstrates that the secretome of osteoblastic cells is more complex than previously reported and supports the emerging evidence that osteoblastic cells secrete proteins with endocrine functions and regulate cellular processes beyond bone formation.

Effects of orbital spaceflight on human osteoblastic cell physiology and gene expression. During long-term spaceflight, astronauts lose bone, in part due to a reduction in bone formation. It is not clear, however, whether the force imparted by gravity has direct effects on bone cells. To examine the response of bone forming cells to weightlessness, human fetal osteoblastic hFOB cells were cultured during the 17 day STS space shuttle mission.

Fashion Victims Vol 2 red vinyl 12". Italian twosome Tale of Us last collaborated with Vaal - London-based daughter of Sting - way back in While that EP contained a couple of much-played tech-house rollers, this latest two-tracker explores different influences. A-side "Monument" is particularly alluring, thanks largely to rising and falling synthesizer arpeggio lines, momentum-shifting percussion, foreboding electronics and a spine-tingling vocal from Vaal.

Flipside "The Hangar" is an altogether more forthright affair. Underpinned by a relentless kick-drum pattern, it's a moody, angular and mind-altering concoction that looks towards industrial techno for inspiration. Becoming N one white vinyl 12". The Full Stream Ahead: Nicolas Jaar's Other People label returns after a brief hiatus with Vaghe Stelle, an Italian producer how is known for his cybernetic approach to electronic music. The mini-LP is a degree view of ambient, noise and electronica, taking the listener to far and distant corners continuously.

Unlike plenty of other abstract electronic releases, Vaghe Stelle has painted a world of his own, one which resembles more of a sci-fi soundtrack than a stand-alone music release. The melodies, pauses, samples, and voices all blend seamlessly to form a new sound that literally sounds a million light years from here.

The Seventh Order white vinyl 12". If you check their discography, there's plenty of hidden gems to uncover. On this record, the enigmatic Valved receives a furious remix treatment from two Juno HQ favourites. The first is a chest-bursting monster from the depths of the drum machines, while the second is a slice sparse and experimental neo-funk with an otherworldly succession of popping kicks and crystal-like background atmospherics.

Expressed heavyweight vinyl 12". Riemann Angels hand-numbered 7" limited to 60 copies. OHM gram vinyl 12". After years spent supporting the underground IDM scene digitally, Glasgow label Ambidextrous makes the leap to vinyl with this killer compilation of ear-catching deep techno and electronica. Christ brings a bubbling range of synth tones to "Rom" before Norken and Nyquist drop some brooding electro tones over rolling beats on"Od Detot". Solipsism has a more sassy house sound to impart, while Nyquist goes into full electro mode on his own.

On the flip, Analogue Audio Association have some edgy acid to throw down, Cyan brings a touch of boogie flex to the record and Mich Chillage rounds the record off with emotive outboard electronics of a reflective nature. Pillow Person Remixes gram vinyl 12". Acid Waxa drew plenty of positive heat for carrying Roy Of The Ravers amongst many other respected braindancers, but now Hot Chip drummer Sarah Jones is getting the remix treatment on the label for her Pillow Person project, with some wild results.

It's great to see Bogdan Raczynski back in action and bringing some gently wonked, emotional acid meanderings to "On Your Way", while Lechuga Zafiro makes an art out of aping footwork, and more specifically "Footcrab" while making it sound like someone just stubbed their toe and got stuck in a loop. Beginning with the 12 minute epic "Flying Kites" which channels early '90s British bleep IDM, until that fast hitting groove hits at just the right time towards the end.

Lush ambient piece "Keep Me Close" works as an effective intermission of sorts on this grainy and saturated dream state captured to VHS. Finally, Venturini displays a more aggressive side as seen on his previous release with the gnarly and slow burning industrial electronics of "Trace Of Smoke". New Army Of Androgynes 12". Black Toms EP 12". Evan Michael , Rumore. Having released over fifty records since their foundation back in , Dark Entries use the widened exposure afforded by that excellent Patrick Cowley compilation released in the last quarter of as a springboard to launch a new dedicated 12" series.

Retaining their archival approach, the first release focuses on the short-lived Italian act Victrola; formed as a four piece combo in Messina, Victrola slimmed down to the synthesizer and guitar-based duo Antonio "Eze" Cuscina and Carlo Smeriglio and moved to the fertile music scene growing in early 80s Florence. In , the pair issued their one record-shaped contribution to the early 80s Italian synth scene in the shape of Maritime Tatami, a two-track 12?

Recorded using the Roland TR and TR at a time when these models had only been made available, so this reissue of Maritime Tatami from Dark Entries offers a chance for people to assess a slice of analogue experimentation at its most nascent. Le Flux De La Nature 10". Voices From The Hell 12". Lee Fraged , Juno Recommends Leftfield , 3. Values 12" in die-cut sleeve. Autumn Equinox Edition limited clear vinyl 12".

KAOS London affiliated label Khemia presents the sounds as heard at their wild and legendary parties on their second release, which follows up great previous editions featuring the links of In Aeternam Vale, Rrose and Orphx.