Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein(RNP)electroporation

Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising cost.Furthermore,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in cattle.In this point of view,electroporation in embryos has been risen as an alternative.Results First,editing efficiency of our electroporation methods were tested for embryos.Presence of mutation on embryo was confirmed by T7E1 assay.With first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,respectively.In case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for BLG.Afterwards,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing.Thirteen recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double KO.Ten surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both genes.Conclusions These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied.Finally,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.

Mechanical and electrical properties of In-Bi solder at Bi_(2212) superconductor interface with annealed Ag spray layers and Ag precoating layers

The electrical properties of solder contact layers between Cu-Ni shunt metal and tube type Bi2212 superconductor that is applied in superconducting fault current limiter were studied.The contact properties of the solders are improved not only by Ag precoating layers,but also by the pre-sprayed Ag layer and subsequent Ag precoating layers.The annealed Ag sprayed layers onto Bi2212 superconductor prior to Ag electroplating work as protecting layers for the superconductor from plating solutions.The contact angle of the electroplated Ag layer is 42.91° and decreases to 15.25° and 5.88° with Ag sprayed layer and additional Ag electroplated layers.The Ag sprayed layer with suitable annealing prior to Ag electroplating improves contact strength of the Ag electroplated layer by about 12% due to denser microstructure of the Ag electroplated layers.

Development of a 3D subcutaneous construct containing insulin-producing beta cells using bioprinting

Type 1 diabetes is caused by insulin deficiency due to the loss of beta cells in the islets of Langerhans.In severe cases,islet transplantation into the portal vein is performed.However,due to the loss of transplanted islets and the failure of islet function,the 5-year insulin independence rate of these patients is<50%.In this study,we developed a long-term,insulin-secreting,3 Dbioprinted construct implanted subcutaneously with the aim of preventing islet loss.The bioprinted construct was fabricated by the multi-layer bioprinting of beta-cell(mouse insulinoma-6:MIN-6)-encapsulated alginate bioink and poly(caprolactone)biodegradable polymer.A glucose response assay revealed that the bioprinted constructs proliferated and released insulin normally during the 4-week in vitro period.Bioprinted MIN-6 generated clusters with a diameter of 100-200μm,similar to the original pancreatic islets in the construct.In an in vivo study using type 1 diabetes mice,animals implanted with bioprinted constructs showed three times higher insulin secretion and controlled glucose levels at 8 weeks after implantation.Because the implanted,bioprinted constructs had a positive effect on insulin secretion in the experimental animals,the survival rate of the implanted group(75%)was three times higher than that of the non-implanted group(25%).The results suggest that the proposed,3 D-bioprinted,subcutaneous construct can be a better alternative to portal vein islet transplantation.

Behavioral inhibition in female college students with schizotypal traits: An event-related potential study

This study investigated behavioral inhibition in female college students with psychometrically defined schizotypal traits using a Go/NoGo task and event-related potentials (ERPs). The schizotypal-trait (n = 15) and normal control (n = 15) groups were selected based on scores of the Schizotypal Personality Questionnaire (SPQ). The Go/NoGo task consisted of Go (requires response) and NoGo (requires no response) conditions. In terms of response time and accuracy rate for the Go/NoGo task, the two groups did not differ significantly. In terms of ERPs, the control group showed greater N2 amplitudes in response to NoGo (NoGo-N2) than to Go stimuli (Go-N2), whereas the schizotypal-trait group showed no significant difference in NoGo-N2 and Go-N2 amplitudes. In addition, the schizotypal-trait group showed reduced NoGo-N2 amplitudes at the frontal site compared to controls, and an association between SPQ scores and NoGo-N2 amplitudes measured at the frontal site. The two groups did not differ in P3 amplitudes. Since the N2 reflects the detection of response conflict and behavioral inhibition, the present results indicate that nonclinical individuals with schizotypal traits have difficulties in detecting response conflict and behavioral inhibition.

nhalation of Sevoflurane and Desflurane Can Not Affect QT Interval, Corrected QT, Tp-Te/QT or Tp-Te/JT in Children

To the Editor： Sevoflurane and desflurane are tile most common general anesthetics used in children. Previous study has shown that these agents can affect the electrocardiogram （ECG） markers,i^i leading to fatal ventricular arrhythmia. The followings were identified as useful indices for predicting and estimating the occurrence of a fatal arrhythmia： the interval between the peak and the end of T wave （Tp-Te） the ratio of the Tp-Te interval to the QT interval （[Tp-Te/QT]）, and the JY interval （JT）, which is measured from the end of the QRS complex to Te.