Fluid flow induced calcium response in bone cell network

In our previous work, bone cell networks with controlled spacing and functional intercellular gap junctions had been successfully established by using microcontact printing and self assembled monolayers technologies [Guo, X. E., E. Takai, X. Jiang, Q. Xu, G. M. Whitesides, J. T. Yardley, C. T. Hung, E. M. Chow, T. Hantschel, and K. D. Costa. Mol. Cell. Biomech. 3:95-107, 2006]. The present study investigated the calcium response and the underlying signaling pathways in patterned bone cell networks exposed to a steady fluid flow. The glass slides with cell networks were separated into eight groups for treatment with specific pharmacological agents that inhibit pathways significant in bone cell calcium signaling. The calcium transients of the network were recorded and quantitatively evaluated with a set of network parameters. The results showed that 18 alpha-GA (gap junction blocker), suramin (ATP inhibitor), and thapsigargin (depleting intracellular calcium stores) significantly reduced the occurrence of multiple calcium peaks, which were visually obvious in the untreated group. The number of responsive peaks also decreased slightly yet significantly when either the COX-2/PGE(2) or the NOS/nitric oxide pathway was disrupted. Different from all other groups, cells treated with 18 alpha-GA maintained a high concentration of intracellular calcium following the first peak. In the absence of calcium in the culture medium, the intracellular calcium concentration decreased slowly with fluid flow without any calcium transients observed. These findings have identified important factors in the flow mediated calcium signaling of bone cells within a patterned network.

Observations on the mating behavior, spawn mass and larval development of Hydatina physis Linne, 1758 (Cephalaspidea: Hydatinidae) from Karachi

Reproduction of Hydatina physis was studied in a population from Karachi, Pakistan, including mating and egg laying behavior, spawn characteristics and development.Individuals first appear in the field in October and remain until March. The spawning occurs from mid-November till mid-February with a peak in December. During this period the individuals were also observed pairing. In captivity, mating lasts for 30 minutes, second mating occurs two days later. Oviposition occurs in a very interesting and unusual manner. The mother turns "up-side-down" with its food fully expanded and the shell completely hidden underneath, the expanded foot serves as protective cover to the eggs. Eggs are deposited in a complexly folded mass with a short stem and an adhesive disc. Capsules, arranged in a single layer, contain 4-6 eggs each of wich is 70 um in diameter. Development is planktotrophic and veligers hatch after 14 days at a temperature of 26-28 degrees Celsius.

Vibration-assisted bone-graft compaction in impaction bone grafting of the femur

The complications of impaction bone grafting in revision hip replacement includes fracture of he femur and subsidence of the prosthesis. In this in vitro study we aimed to investigate whether the use of vibration, combined with a perforated tamp during the compaction of morsellised allograft would reduce peak loads and hoop strains in the femur as a surrogate marker of the risk of fracture and whether it would also improve graft compaction and prosthetic stability. We found that the peak loads and hoop strains transmitted to the femoral cortex during graft compaction and subsidence of the stem in subsequent mechanical testing were reduced. This innovative technique has the potential to reduce the risk of intra-operative fracture and to improve graft compaction and therefore prosthetic stability. © 2007 British Editorial Society of Bone and Joint Surgery.

Land-based mass production of prawn (Penaeus monodon Fabricius) spawners

The first spawnings were obtained 12 days after ablation with 4 spawners yielding 784,000 eggs and a harvest of 250,000 P SUB-10 fry. Survival of females after 1 month was approximately 30%. Mortalities were mostly due to handling stress during the regular ovarian samplings as well as disease frm the accumulated excess feeds on the bottom of the tank. Male survival could not be recorded because of transfers to other tanks and addition of new stocks. Development seemed to peak 3 weeks after ablation. The average number of eggs per ablated spawner was 120,000. However, many of the partially spawned females were removed from the spawning tanks the following day so that remaining eggs released in the next 2 to 3 days could not be recorded. Estimate of the average number of eggs per ablated spawner is 120,000-150,000 in contrast to 500,000 per wild spawner. However, the low production cost more than compensates for the difference. Fry reared in the Wet Laboratory were used for experiments, mostly on feeding. Therefore, survival at harvest is not to be taken as a reflection of stock quality. Although fewer in number, larvae from ablated prawns are as healthy in terms of vigor in swimming and feeding as those from wild females. Most mortalities are due to inability to molt caused by lower water temperatures and inadequate feeding.

Computer assisted peak recognition for polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) in environmental samples

A software has been developed for the peak recognition of 136 polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) after high resolution gas chromatography coupled with mass spectrometry (HRGC/HRMS). Based on the retention times of C-13 labelled 2,3,7,8-substituted PCDD/F internal standards, the retention times of all PCDD and PCDF can be calibrated automatically and accurately. Therefore, it is very convenient to identify the peaks by comparing the retention of samples and the calibrated retention times of their chromatograms. Hence, this approach is very significant because it is impossible to obtain always a standard chromatogram and PCDD/F analysis are very expensive and time consuming. The calibration results can be transferred to Excel for calculation. The approach is a first step to store costly and environmentally relevant data for future application.

(Ga,Mn,N) compounds growth with mass-analyzed low energy dual ion beam deposition

The (Ga,Mn,N) samples were grown by the implantation of low-energy Mn ions into GaN/Al2O3 substrate at different elevated substrate temperatures with mass-analyzed low-energy dual ion beam deposition system. Auger electron spectroscopy depth profile of samples grown at different substrate temperatures indicates that the Mn ions reach deeper in samples with higher substrate temperatures. Clear X-ray diffraction peak from (Ga,Mn)N is observed in samples grown at the higher substrate temperature. It indicates that under optimized substrate temperature and annealing conditions the solid solution (Ga,Mn)N phase in samples was formed with the same lattice structure as GaN and different lattice constant. (C) 2003 Elsevier Science B.V. All rights reserved.

GdxSi grown with mass-analyzed low energy dual ion beam epitaxy technique

Semiconducting gadolinium silicide GdxSi samples were prepared by mass-analyzed low-energy dual ion beam epitaxy technique. Auger electron spectroscopy depth profiles indicate that the gadolinium ions are implanted into the single-crystal silicon substrate and formed 20 nm thick GdxSi film. X-ray double-crystal diffraction measurement shows that there is no new phase formed. The XPS spectra show that one type of silicon peaks whose binding energy is between that of silicide and silicon dioxide, and the gadolinium peak of binding energy is between that of metal Gd and Gd2O3. All of these results indicate that an amorphous semiconductor is formed. (C) 2002 Elsevier Science B.V. All rights reserved.

Interactions of mitoxantrone with duplex and triplex DNA studied by electrospray ionization mass spectrometry

We have examined interactions between mitoxantrone (MXT) and DNA duplexes or triplexes with different base compositions by using electrospray ionization mass spectrometry (ESI-MS), respectively. MXT interacts preferentially with DNA duplexes compared to the triplexes. In the mass spectrum of the duplex-MXT mixture, the complex peaks dominated in the ratios of duplex/MXT of 1:1, 1:2 and 1:3, and the 1:2 duplex/MXT peak was the most abundant. In contrast, only 1:1 triplex-MXT complexes were observed in the mass spectrum of the triplex-MXT mixture, and the most intensive peak was a free triplex ion without MXT.

Microstructures and mechanical properties of Mg-8Gd-0.6Zr-xNd (x=0, 1, 2 and 3 mass%) alloys

Mg-8Gd-0.6Zr-xNd (x = 0, 1, 2 and 3 mass%) alloys were prepared by metal mould casting method, and the microstructures, age hardening responses and mechanical properties have been investigated. The microhardness of the as-cast alloys is increased with increasing Nd content. The age hardening behavior and mechanical properties are enhanced significantly by adding Nd element. The peak ageing hardness of the Mg-8Gd-0.6Zr-3Nd alloy is 103, it is about 1.3 times more than that of the Mg-8Gd-0.6Zr alloy. The aged Mg-8Gd-0.6Zr-3Nd alloy exhibits maximum ultimate tensile strength and yield strength, and the values are 271 and 205 MPa at room temperature, 205 MPa and 150 MPa at 250 degrees C, respectively. Which are about 2 times higher than those of Mg-8Gd-0.6Zr alloy. The improved hardness and strength are mainly attributed to the fine dispersiveness Of Mg5RE and Mg12RE precipitates in the alloy.

Characterization of non-covalent complexes of rutin with cyclodextrins by electrospray ionization tandem mass spectrometry

Electrospray ionization tandem mass spectrometry (ESI-MSn) and the phase solubility method were used to characterize the gas-phase and solution-phase non-covalent complexes between rutin (R) and alpha-, beta- and gamma-cyclodextrins (CDs). The direct correlation between mass spectrometric results and solution-phase behavior is thus revealed. The order of the 1:1 association constants (K-c) of the complexes between R and the three CDs in solution calculated from solubility diagrams is in good agreement with the order of their relative peak intensities and relative collision-induced dissociation (CID) energies of the complexes under the same ESI-MSn condition in both the positive and negative ion modes. Not only the binding stoichiometry but also the relative stabilities and even binding sites of the CD-R complexes can be elucidated by ESI-MSn. The diagnostic fragmentation of CD-R complexes, with a significant contribution of covalent fragmentation of rutin leaving the quercetin (Q) moiety attached to the CDs, provides convincing evidence for the formation of inclusion complexes between R and CDs. The diagnostic fragment ions can be partly confirmed by the complexes between Q and CDs. The gas-phase stability order of the deprotonated CD-R complexes is beta-CD-R > alpha-CD-R > gamma-CD/R; beta-CD seems to bind R more strongly than the other CDs.

Characterization of triterpenoidic saponin mixture in crude extracts from leaves of Acanthopanax senticosus Harms by saponin structural correlation and mass spectrometry

Eighteen triterpenoidic saponins in crude extracts from leaves of Acanthopanax senticous Harms have been investigated by electrospray ionization multi-stage tandem mass spectrometry and high-resolution mass spectrometry. In ESI-MS spectra, predominant [M + Na](+) ions in the positive ion mode have been observed for molecular mass information. Meanwhile, specific structural correlations between these ions are firstly found. The 18 peaks (ions) can be classified into three groups (group D, E, and F with mass increase) with each group including six peaks. There is a mass difference of 132 Da between group D and E for each corresponding peak in turn (for example peak 1 to peak 7), indicating one more pentose residue was attached to saponins in group E than those corresponding in group D. The mass difference of 146 Da between group E and F implies one more deoxy-hexose attached to saponins in group F than those corresponding in group E. The structural correlations of the corresponding ions are confirmed by tandem mass spectrometry and high-re solution mass spectrometry. These structural features can not only facilitate the rapid characterization of the native known saponins in crude plant extracts, thus avoiding tedious derivation and separation of saponins, but also help find novel compounds of the same type in a specific medicinal plant.

Microstructures and tensile properties of Mg-8Gd-0.6Zr-xNd-yY (x+y=3, mass%) alloys

Mg-8Gd-0.6Zr-xNd-yY (mass%) alloys which containing different Nd:Y mass ratio of 3:0, 2:1, 1:2 and 0:3 with a constant x + y = 3 were prepared by metal mould casting method, and the microstructure, aging behaviour and tensile properties have been investigated. The fibrous eutectic areas along the boundaries enlarge clearly in the as-cast alloys containing Y element, and the fine grain boundaries and dispersed precipitation are observed in the aged alloys. The Mg-8Gd-0.6Zr-2Nd-Y alloy exhibits notably age-hardening behaviour and the highest mechanical property. The ultimate tensile strength and yield strength of Mg-8Gd-0.6Zr-2Nd-Y alloy in the peak aged hardness are 293 and 221 MPa at room temperature, 248 and 191 MPa at 230 degrees C. The improvement of age-hardening response and tensile properties is mainly attributed to the quadrate-like stable Mg5RE precipitate, which forms readily and orderly in aged Mg-8Gd-0.6Zr-2Nd-Y alloy.

Studies on the non-covalent complexes between oleanolic acid and cyclodextrins using electrospray ionization tandem mass spectrometry

Non-covalent inclusion complexes formed between an anti-inflammatory drug, oleanolic acid (OA), and alpha-, beta- and gamma-cyclodextrins (CDs) were investigated by means of solubility studies and electrospray ionization tandem mass spectrometry (ESI-MSn). The order of calculated association constants (K-1:1) of complexes between OA and different CDs in solution is in good agreement with the order of their relative peak intensities and the relative CID energies of the complexes under the same ESI-MSn conditions. These results indicate a direct correlation between the behaviors of solution- and gas-phase complexes. ESI-MS can thus be used to evaluate solution-phase non-covalent complexes successfully. The experimental results show that the most stable 1:1 inclusion complexes between three CDs and OA can be formed, but 2:1 CD-OA complexes can be formed with beta- and gamma-CDs. Multi-component complexes of alpha-CD-OA-beta-CD (1:1:1), alpha-CD-OA-gamma-CD (1:1:1) and beta-CD-OA-gamma-CD (1:1:1) were found in equimolar CD mixtures with excess OA. The formation of 2:1 and multi-component 1:1:1 non-covalent CD-OA complexes indicates that beta- and gamma-CD are able to form sandwich-type inclusion non-covalent complexes with OA. The above results can be partly supported by the relative sizes of OA and CD cavities by molecular modeling calculations.

Capillary liquid chromatographic-high-resolution mass spectrometric analysis of ribonucleotides

A method of capillary HPLC-high-resolution MS was developed for the trace analysis of ATP, GTP, dATP and dGTP Dimethylhexylamine (DMHA) was used as ion-pairing agent for the HPLC retention and separation of the nucleotides and positive ion electrospray time-of-flight MS was used for the detection. The application of capillary HPLC allowed minimal usage of DMHA while providing excellent peak retention and resolution, which significantly reduced the ion suppression in electrospray ionization-MS analysis and thus increased the sensitivity. Adduct ions of nucleotides and DMHA were used as quantitative ions in order to achieve the best sensitivity. DMHA concentration at 5 mM in the aqueous mobile phase at pH 7 was found to be the optimal conditions for the C Is capillary column. The method was applied to determine ATP level in cultured C6 glioma cells that were treated with toxic concentrations of Zn. The results showed that the cellular ATP level decreased from 2.7 pmol/cell (<10% cell death) in average control cell samples to 0.36 pmol/cell as the concentration of Zn increased to 120 mg/l (>35% cell death) in culture medium.

Analysis of chemical constituents of essential oil in purple magnolia leaves by gas chromatography-mass spectrometry

The essential oil in purple magnolia leaves was extracted by steam distillation approaches. The oil obtained was dried with anhydrous magnesium sulfate. According to the analysis of gas chromatography/mass spectrometry, more than 40 peaks were separated and 32 compounds were identified. The identified constituents represent 95% of the peak area of the essential oil. The main compounds were germacrene-D, santolina triene, caryophyllene, 1,3,7-octatriene, 3,7-dimethyl, and camphene, etc.