The objective of this study was to determine the potential hazardous effects of sorting process by flowcytometry on the quality of boar spermatozoa by flowcytometer. Freshly collected boar semen was diluted and divided into two groups; control none sorted and sorted. Sperms in sorted group were processed with flowcytometer for cell sorting with 100 uM nozzle under the 20 psi pressure. Measurements on each parameter were made at two time points, 0hr (right after sorting) and 24 hr post sorting. Although there was a tendency of lower viability in sorted group than none sorted control group, the percentage of live cells in control(75.83+-6.92 & 59.53+-10.34) was not significantly different from sorted (59.7+-7.34 & 43.97+-3.76) at both 0 and 24 hr post sorting. However, sorted sperm showed significantly lower mitochondrial function compared to the control at both 0 h (79.37+-3.22 vs. 63.50+-10.05) and 24hr(67.27+-3.22 vs. 46.97+-5.37) time points (p<0.007). Sperm DNA fragmentation rate was significantly lower in control (22.0+-7.04) than that of sorted (32.27+-7.49) at 24 hr time point (p<0.0002). Taken together, these data suggested thatsorting process by flowcytometer may have influenced sperm motility rather than viability. Also high speed sperm sorting by flowcytometer has significant effects on DNA fragmentation on elapsed time after sorting.
A quantitative sequencing (QS) protocol that detects the frequencies of sodium channel mutations (M815I, T917I and L920F) responsible for knockdown resistance in permethrin-resistant head lice was tested as a population genotyping method. Genomic DNA fragments of the sodium channel α-subunit gene that encompass the three mutation sites were PCR-amplified from individual head lice with either resistant or susceptible genotypes, and combined together in various ratios to generate standard DNA template mixtures for QS. Following sequencing, the signal ratios between resistant and susceptible nucleotides were calculated and plotted against the corresponding resistance allele frequencies. Quadratic regression coefficients of the plots were close to 1, demonstrating that QS is highly reliable for the prediction of resistance allele frequencies. Prediction of resistance allele frequencies by QS in several globally collected lice samples including 12 Korean lice populations suggested that permethrin resistance varied substantially amongst different geographical regions. Three local populations of Korean lice were determined to have 9.8-36.7% resistance allele frequencies, indicating that an urgent resistance management is needed. QS should serve as a preliminary resistance monitoring tool for proper management strategies by allowing early resistance detection.
Four species in two genera of the tribe Pilophorini, the ant mimetic Plant bugs are revised from Nepal. One new species of genus Pilophorus Hann, P. sp. nov is proposed. Pilophorus typicus (Distant, 1909) and Sthenaridea piceonigra (Motschulsky, 1863) are new record to Nepal. Pilophours josifovianus Duwal and Yasunaga, 2008 previously described species from Nepal and all other three species are here with illustrations of male and female genitalia and their biological information.
Spectral line profiles of filaments/prominences to be observed by the Fast Imaging Solar Spectrograph (FISS) are studied. The main spectral lines of interests are Hα Ca II 8542, and Ca II K. FISS has a high spectral resolving power of 2 x 105, and supports simultaneous dual-band recording. This instrument will be installed at the 1.6m New Solar Telescope (NST) of Big Bear Solar Observatory, which has a high spatial resolution of 0.065" at 500nm. Adopting the cloud model of radiative transfer and using the model parameters inferred from pre-existing observations, we have simulated a set of spectral profiles of the lines that are emitted by a filament on the disk or a prominence at the limb. Taking into account the parameters of the instrument, we have estimated the photon count to be recorded by the CCD cameras, the signal-to-noise ratios, and so on. We have also found that FISS is suitable for the study of multi-velocity threads in filaments if the spectral profiles of Ca II lines are recorded together with Hα lines
Over the past two decades a number of methodologies have been developed to efficiently evaluate seismic performance of existing buildings, which are briefly reviewed here. Building upon previous methodologies and experience using them as well as new lessons learned from earthquakes, FEMA-356 Prestandard and Commentary for the Seismic Rehabilitation of Buildings, is the latest method which is reviewed in some what more in depth in this paper. FEMA-356 methodology uses performance-based engineering concept and provides the user with means to evaluate buildings for various rehabilitation objectives. Rehabilitation objectives are based on building performance levels - such as collapse prevention, life safety, immediate occupancy, and operational levels - and earthquake hazard levels. The methodology addresses building performance according to the performance of both the building structural elements and as well as nonstructural components. The structural elements and components are grouped to primary and secondary components. The primary components are those that provide the capacity of structure to resist collapse and secondary components are all other components that affect the lateral stiffness or distribution of forces in the structure. The primary and secondary components are evaluate dusing different acceptance. The hazard may be defined via published hazard maps and standard response spectrum fOTIns or via site-specific earthquake hazard assessment. The application of the FEMA-356 is demonstrated here by evaluating an existing concrete frame with steel roof building.
We have previously produced transgenic (TG) mice expressing the human lactoferrin (hLF), interleukin-10 (hIL-10), and thrombopoietin (hTPO) proteins in the milk. In this study, we examined whether simple crossbreeding between two kids of a single transgenic mouse can produce double transgenics co-expressing two human proteins.. The hLF male, and the hIL-10 male were crossbred with the hIL-10 and hTPO females, and the hTPO female, respectively. PCR analysis for genotyping showed 32%, 23% and 24% double transgenic rates for hLF/hIL-10, hLF/hTPO, and hIL-10/hTPO transgenes, respectively. We analyzed the expression levels of the human proteins from double transgenic mice and compared those with their single transgenic siblings. All double transgenic co-expressed two human proteins at comparable levels to singles', unless hTPO was not co-expressed: for hLF, 1.1 mg/ml in hLF/hIL-10, whereas 0.5 mg/ml in hLF/hTPO; for hIL-10, 4.1 mg/ml in hIL-10/hLF, whereas 1.4 mg/ml in hIL-10/hTPO. Ihe downregulation of hTPO to half level of singles' was observed in double transgenic mice. The possible reason why hTPO co-expressed might lead to down-regulation of another human protein was discussed. These results suggested that double transgenic generated by crossbreeding between two singles' could be useful system for bioreactor.
Most studies on transgenic bioreactors have focused on expression levels of interest genes. In this study we examined whether transgenic bioreactors would inherit expression level of the Oansgene to long-term generations independently of transgene sources. We employed three transgenic mice, which were separately reported, carrying different transgenes and copy numbers, 27 kb of hLF and 22 kb of hIL-10 genomic sequences, and 1.3 kb of hTPO cDNA, respectively. Three females of the transgenic lineages crossbred with a wild-type male up to 20 generations to test transgenic frequencies of their progenies and to determine expression levels of the transgenes. Ultimately, transmission rates of kLF, hIL-10, and hTPO were 64.3+-7.0, 59.3+-9.8, and 56.1+-9.7, respectively, appeared following Mendelian pattern of inheritance. Notably, we found that levels of expressions of hLF, hIL-10, and hTPO in milk were sustained to high numbers of generations. No transgene silencing of expression was observed in every generations of all transgenic mice. In conclusion, we suggest that once established animal bioreactors could consistently transmit the transgene to continual generations, without loss of expressional activity, independently of transgene sources.
The ability of cow dung ash without any pretreatment to remove color from textile dyes N Blue RGB, Green B and EOSIN YWS from aqueous solution has been investigated in this work. Cow dung ash, an ecofriendly and low cost adsorbent was prepared by burning cow dung cakes in the muffle furnace at 500℃. The adsorption was achieved under different pH and adsorbate concentration. The data was fitted to simple polynomial and the isotherms similar to Langmuir and Freundlich isotherms.