Improved efficiencies in the generation of multigene-modified pigs by recloning and using sows as the recipient

Zygote ◽  
2021 ◽  
pp. 1-8
Author(s):  
Jongki Cho ◽  
Ghangyong Kim ◽  
Ahmad Yar Qamar ◽  
Xun Fang ◽  
Pantu Kumar Roy ◽  
...  
Keyword(s):  
Production Efficiency ◽  
Developmental Competence ◽  
Cell Group ◽  
Fetal Cell ◽  
Endothelial Protein C Receptor ◽  
Transgenic Pigs ◽  
Increase Production Efficiency ◽  
Fetal Cells ◽  
Donor Cells ◽  
Cell Groups

Summary This study was performed to improve production efficiency at the level of recipient pig and donor nuclei of transgenic cloned pigs used for xenotransplantation. To generate transgenic pigs, human endothelial protein C receptor (hEPCR) and human thrombomodulin (hTM) genes were introduced using the F2A expression vector into GalT –/– /hCD55 + porcine neonatal ear fibroblasts used as donor cells and cloned embryos were transferred to the sows and gilts. Cloned fetal kidney cells were also used as donor cells for recloning to increase production efficiency. Pregnancy and parturition rates after embryo transfer and preimplantation developmental competence were compared between cloned embryos derived from adult and fetal cells. Significantly higher parturition rates were shown in the group of sows (50.0 vs. 4.1%), natural oestrus (20.8 vs. 0%), and ovulated ovary (16.7 vs. 5.6%) compared with gilt, induced and non-ovulated, respectively (P < 0.05). When using gilts as recipients, final parturitions occurred in only the fetal cell groups and significantly higher blastocyst rates (15.1% vs. 21.3%) were seen (P < 0.05). Additionally, gene expression levels related to pluripotency were significantly higher in the fetal cell group (P < 0.05). In conclusion, sows can be recommended as recipients due to their higher efficiency in the generation of transgenic cloned pigs and cloned fetal cells also can be recommended as donor cells through correct nuclear reprogramming.

Effects of donor sex and different cell treatments on development of yak–bovine interspecies nuclear transfer embryos

2008 ◽  
Vol 5 (1) ◽  
pp. 55-60
Author(s):  
Liu Ying ◽  
Zhu Shi-En ◽  
Li Rong ◽  
Wang Li-Li ◽  
Wang Hai-Ping ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Nuclear Transfer ◽  
Developmental Competence ◽  
Cell Group ◽  
Serum Starvation ◽  
Cleavage Rate ◽  
Cell Cycle Synchronization ◽  
Donor Cells ◽  
Significant Difference ◽  
Interspecies Nuclear Transfer

AbstractThe purpose of this study was to evaluate the effects of donor sex, treatments of cell cycle synchronization and donor nuclei obtained from fresh or frozen–thawed conditions on developmental competence of yak–bovine interspecies nuclear transfer embryos. Bovine (Bos taurus) oocytes were used as recipients and yak (Bos grunniens) ear fibroblast cells were used as donors. Results indicated that the development rate of male blastocysts was higher than that of female (56.6% versus 39.5%, P<0.05), whereas cleavage and total cell number showed no difference between the two groups. No significant difference was observed in the development and quality of blastocysts with donor cells treated by serum starvation or contact inhibition, and there was no significant difference in embryo development with fresh or frozen–thawed donor cells, whereas the cleavage rate in the group of frozen–thawed cells was significantly lower than that of the fresh cell group (54.5% versus 78.2%, P<0.05). The results demonstrated that donor sex could impact the developmental competence of yak–bovine interspecies nuclear transfer embryos, whereas different treatments of cell cycle synchronization and freezing had little influence.

scholarly journals Development of Immunohistochemical Methods for Casein Detection in Meat Products

Foods ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 28
Author(s):  
Ludmila Kalčáková ◽  
Matej Pospiech ◽  
Bohuslava Tremlová ◽  
Zdeňka Javůrková ◽  
Irina Chernukha
Keyword(s):  
Milk Protein ◽  
Production Efficiency ◽  
Raw Materials ◽  
Meat Products ◽  
Milk Proteins ◽  
Increase Production Efficiency ◽  
Allergenic Potential ◽  
Significant Difference ◽  
Dairy Ingredients ◽  
Immunohistochemical Methods

To increase production efficiency of meat products, milk protein additives are often used. Despite a number of advantages, use of dairy ingredients involves a certain risk, namely the allergenic potential of milk proteins. A number of methods have been developed to detect milk-origin raw materials in foodstuffs, including immunological reference methods. This study presents newly developed immunohistochemical (IHC) methods for casein detection in meat products. Casein was successfully detected directly in meat products where sensitivity was determined at 1.21 and specificity at 0.28. The results obtained from the IHC were compared with the Enzyme-Linked Immuno Sorbent Assay (ELISA) and there was no statistically significant difference between the IHC and ELISA methods (p > 0.05). The correspondence between the methods was 72% in total. The highest correspondence was reached in frankfurters (90%), the lowest in canned pâté (44%).

scholarly journals Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells

2021 ◽  
Vol 22 (3) ◽  
pp. 1386
Author(s):  
Jerzy Wiater ◽  
Marcin Samiec ◽  
Maria Skrzyszowska ◽  
Daniel Lipiński
Keyword(s):  
Genetic Resource ◽  
Ex Vivo ◽  
Trichostatin A ◽  
Expression Profiles ◽  
Fibroblast Cells ◽  
Control Groups ◽  
Transgenic Pigs ◽  
Cell Tissue ◽  
Donor Cells ◽  
Lower Expression

This study was conducted to explore whether trichostatin A-assisted epigenomic modulation (TSA-EM) can affect the expression of not only recombinant human α1,2-fucosyltransferase (rhα1,2-FT) and α-galactosidase A (rhα-Gal A) immune system enzymes but also Galα1→3Gal epitopes in ex vivo proliferating adult cutaneous fibroblast cells (ACFCs) derived from hFUT2×hGLA bi-transgenic pigs that had been produced for the needs of future xenotransplantation efforts. The ACFC lines were treated with 50 nM TSA for 24 h and then the expression profiles of rhα1,2-FT and rhα-Gal A enzymes were analyzed by Western blot and immunofluorescence. The expression profiles of the Galα1→3Gal epitope were determined by lectin blotting and lectin fluorescence. The ACFCs derived from non-transgenic (nTG) pigs were served as the negative (TSA−) and positive (TSA+) control groups. For both hFUT2×hGLA and nTG samples, the expression levels of α1,2-FT and α-Gal A proteins in TSA+ cells were more than twofold higher in comparison to TSA− cells. Moreover, a much lower expression of the Galα1→3Gal epitopes was shown in TSA− hFUT2×hGLA cells as compared to the TSA− nTG group. Interestingly, the levels of Galα1→3Gal expression in TSA-treated hFUT2×hGLA and nTG ACFCs were significantly higher than those noticed for their TSA-untreated counterparts. Summing up, ex vivo protection of effectively selected bi-transgenic ACFC lines, in which TSA-dependent epigenetic transformation triggered the enhancements in reprogrammability and subsequent expression of hFUT2 and hGLA transgenes and their corresponding transcripts, allows for cryopreservation of nuclear donor cells, nuclear-transferred female gametes, and resultant porcine cloned embryos. The latter can be used as a cryogenically conserved genetic resource of biological materials suitable for generation of bi-transgenic cloned offspring in pigs that is targeted at biomedical research in the field of cell/tissue xenotransplantation.

scholarly journals Pigment Production under Cold Stress in the Green Microalga Chlamydomonas reinhardtii

Agriculture ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 564
Author(s):  
Supakorn Potijun ◽  
Chonlada Yaisamlee ◽  
Anchalee Sirikhachornkit
Keyword(s):  
Cold Stress ◽  
Chlamydomonas Reinhardtii ◽  
Production Efficiency ◽  
Cell Physiology ◽  
Efficient Production ◽  
Increase Production Efficiency ◽  
Whole Cells ◽  
Stress Acclimation ◽  
Chlorophyll Accumulation ◽  
In The Wild

Microalgae have long been used for the commercial production of natural colorants such as carotenoids and chlorophyll. Due to the rising demand for carotenoids and other natural products from microalgae, strategies to increase production efficiency are urgently needed. The production of microalgal biorefineries has been limited to countries with moderate climates. For countries with cooler climates and less daylight, methodologies for the efficient production of microalgal biorefineries need to be investigated. Algal strains that can be safely consumed as whole cells are also attractive alternatives for developing as carotenoid supplements, which can also contain other compounds with health benefits. Using such strains helps to eliminate the need for hazardous solvents for extraction and several other complicated steps. In this study, the mesophilic green alga Chlamydomonas reinhardtii was employed to study the effects of cold stress on cell physiology and the production of pigments and storage compounds. The results showed that temperatures between 10 and 20 °C induced carotenoid and chlorophyll accumulation in the wild-type strain of C. reinhardtii. Interestingly, the increased level of carotenoids suggested that they might play a crucial role in cold stress acclimation. A temperature of 15 °C resulted in the highest carotenoid and chlorophyll productivity. At this temperature, carotenoid and chlorophyll productivity was 2 times and 1.3 times higher than at 25 °C, respectively. Subjecting a mutant defective in lutein and zeaxanthin accumulation to cold stress revealed that these two carotenoids are not essential for cold stress survival. Therefore, cold temperature could be used as a strategy to induce and increase the productivity of pigments in C. reinhardtii.

A Friction Stir Welding Platform of Thin Plates

2012 ◽  
Vol 628 ◽  
pp. 206-210 ◽  
Author(s):  
Jia Liang Zhang ◽  
Bei Zhi Li ◽  
Xin Chao Zhang ◽  
Qing Xia Wang
Keyword(s):  
Friction Stir Welding ◽  
Virtual Instrument ◽  
Production Efficiency ◽  
Cost Effective ◽  
Field Analysis ◽  
Sensor Data ◽  
Thin Plates ◽  
Increase Production Efficiency ◽  
Friction Stir ◽  
Welding Processes

Friction stir welding processes involve many variables. Engineers and operators often find it difficult to effectively design or control it. The objective of this work is to develop a friction stir welding platform of thin plates to improve welding quality and to increase production efficiency. The study is conducted by using finite element modeling and temperature field analysis technology to obtain optimization parameters, and using virtual instrument, multi-sensor data fusion to monitor the force of the stirring spindle. Experiment results show that the developed platform can reach the requirements of processing quality and is cost-effective.

Identification of Basolateral Amygdala Projection Cells and Interneurons Using Extracellular Recordings

2006 ◽  
Vol 96 (6) ◽  
pp. 3257-3265 ◽  
Author(s):  
Ekaterina Likhtik ◽  
Joe Guillaume Pelletier ◽  
Andrei T. Popescu ◽  
Denis Paré
Keyword(s):  
Firing Rate ◽  
Basolateral Amygdala ◽  
Cell Group ◽  
Small Subset ◽  
Cortical Projection ◽  
Antidromic Activation ◽  
Extracellular Recordings ◽  
Firing Rates ◽  
Cell Groups ◽  
Exact Position

This study tested whether firing rate and spike shape could be used to distinguish projection cells from interneurons in extracellular recordings of basolateral amygdala (BLA) neurons. To this end, we recorded BLA neurons in isoflurane-anesthetized animals with tungsten microelectrodes. Projection cells were identified by antidromic activation from cortical projection sites of the BLA. Although most projection cells fired spontaneously at low rates (<1 Hz), an important subset fired at higher rates (up to 6.8 Hz). In fact, the distribution of firing rates in projection cells and unidentified BLA neurons overlapped extensively, even though the latter cell group presumably contains a higher proportion of interneurons. The only difference between the two distributions was a small subset (5.1%) of unidentified neurons with unusually high firing rates (9–16 Hz). Similarly, distributions of spike durations in both cell groups were indistinguishable, although most of the fast-firing neurons had spike durations at the low end of the distribution. However, we observed that spike durations depended on the exact position of the electrode with respect to the recorded cell, varying by as much as 0.7 ms. Thus neither firing rate nor spike waveform allowed for unequivocal separation of projection cells from interneurons. Nevertheless, we propose the use of two firing rate cutoffs to obtain relatively pure samples of projection cells and interneurons: ≤1 Hz for projection cells and ≥7 Hz for fast-spiking interneurons. Supplemented with spike-duration cutoffs of ≥0.7 ms for projection cells and ≤0.5 ms for interneurons, this approach should keep instances of misclassifications to a minimum.

scholarly journals Specifically Binding of D-Amino Neuraminic Acid to Ganglioside Studied in Prostate Cancer Cells

2021 ◽  
Author(s):  
Kenan Demir ◽  
Huseyin Aktug ◽  
Gurkan Yigitturk ◽  
Eda Acikgoz ◽  
Gunnur Guler ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Enzyme Inhibitor ◽  
Cancer Cell Line ◽  
Prostate Cancer Cell Line ◽  
Cell Group ◽  
Synthesis Inhibitor ◽  
Cell Groups ◽  
Negative Effect ◽  
Synthase Inhibitor

Abstract Aims: The aim of this study was to investigate the cellular binding site of human KDN (2-keto-3-deoxy-D-glycero-D-galacto-nononic acid). The KDN molecule is a member of the sialic acid family, and its expression increases in cancer cells. Although KDN has been shown to bind to GM3 (Monosialodihexosyl Ganglioside) in trout sperm.Methods and Results: Prostate cancer cell line (DU145) was used in this study. Each experimental group was divided into 3 groups: control, GCS (Glucosylceramide synthase) enzyme inhibitor Genz-123346 treated, and GM3 synthesis inhibitor Triptolide treated. Each group was stained using the immunocytochemical method for GM3, GD3 (Disialosyllactosylceramide) and KDN. The FTIR (Fourier Transform Infrared Spectroscopy) analysis was performed to elucidate the cellular changes upon treatment. The non-treated number 1 cell group stained positive with all of GM3, GD3 and KDN, the GCS enzyme blocked with Genz-123346 number 2 cell groups stained positive with only KDN. Furthermore, GD3 Synthase inhibitor Triptolide treated number 3 cell group stained positive with GM3 and KDN. Measurements with FTIR showed apoptotic features with Triptolide while Genz-123346 had no negative effect on the cell viability. The decrease in sugar constructions was revealed and the results that we obtained with staining were reinforced.Conclusions: Determining the location of bounded KDN is important in selecting new targets for cancer treatment researches. It has been shown that KDN is not inhibited by both GM3 inhibition and GD3 inhibition, and thus, KDN might also bind to different places, be specific, and not only attached to any of gangliosides of the GM or GD series.

scholarly journals Perbaikan Kemasan Kerupuk Singkong (Samiler) di UKM Karya Lestari Jaya Tulungagung

2020 ◽  
Vol 4 (1) ◽  
pp. 9
Author(s):  
Dadang Hermawan ◽  
Andy Hardianto ◽  
Arief Rizki Fadhillah
Keyword(s):  
Manihot Esculenta ◽  
Production Efficiency ◽  
Production Capacity ◽  
Raw Material ◽  
Manihot Esculenta Crantz ◽  
Increase Production Efficiency ◽  
Packaging Process ◽  
The People ◽  
Continuous Band ◽  
Break Even Point

Cassava or cassava (Manihot esculenta Crantz) is one of Indonesia's local carbohydrate sources which ranks third largest after rice and corn. One of the uses of cassava is cassava crackers (samiler). Cassava crackers (samiler) are very popular snacks among the people, made from cassava, thin round in shape and taste delicious. Production Capacity of Karya Lestari Jaya's UKM in producing cassava crackers (samiler) in 1 day is ± 20 kg of material or 5 kg of dried cassava crackers (samiler). In one month cassava cracker orders (samiler) can reach ± 80-120 kg. The production process of cassava crackers (samiler) goes through several stages, including: raw material process, printing process, steam process, drying process and packaging process. The problem that can be identified in the field of production is the limited equipment in the packaging process and weighing the samiler cassava crackers. Karya Lestari Jaya's UKM partners in calculating product prices per package have not met the break even point (BEP) standard, so it cannot be maximized in business management. From the above conditions, the method and results of this service are providing technology transfer by providing vaccum sealer machines, continuous band sealers, and digital scales as well as training in the use of tools and training in Break Even Point (BEP) so as to increase production efficiency of cassava crackers (samiler). ABSTRAKSingkong atau ubikayu (Manihot esculenta Crantz) merupakan salah satu sumber  karbohidrat lokal Indonesia yang menduduki urutan ketiga terbesar setelah padi dan jagung. Salah satu pemanfaatan singkong adalah kerupuk singkong (samiler).  Kerupuk singkong (samiler) adalah makanan ringan yang sangat populer dikalangan masyarakat, terbuat dari singkong , berbentuk bundar tipis dan rasanya gurih. Kapasitas Produksi UKM Karya Lestari Jaya dalam memproduksi kerupuk singkong (samiler) dalam 1 hari sebanyak ± 20 kg bahan atau 5 kg kerupuk singkong (samiler) kering. Dalam satu bulan pesanan kerupuk singkong (samiler) dapat mencapai ± 80-120 kg. Proses Produksi kerupuk singkong (samiler) melalui beberapa tahapan, antara lain : proses bahan baku, proses pencetakan, proses steam, proses penjemuran dan proses pengemasan. Permasalahan yang dapat diidentifikasi dalam bidang produksi adalah keterbatasan peralatan dalam proses packaging dan penimbangan kerupuk singkong samiler. Mitra UKM Karya Lestari Jaya dalam melakukan perhitungan harga produk per kemasan belum memenuhi standar break even point (BEP), sehingga tidak dapat maksimal dalam pengelolaan usaha. Dari kondisi diatas maka metode dan hasil dari pengabdian ini adalah memberikan transfer teknologi dengan memberikan Mesin vaccum sealer, continuous band sealer, dan timbangan digital serta melakukan pelatihan penggunaan alat dan pelatihan Break Even Point (BEP) sehingga meningkatkan efisiensi produksi Kerupuk singkong (samiler).

Efficient ruminant production from grassland

2009 ◽  
Vol 2009 ◽  
pp. 248-248
Author(s):  
N D Scollan ◽  
J Moorby
Keyword(s):  
Nitrous Oxide ◽  
Production Efficiency ◽  
Anthropogenic Sources ◽  
Nitrous Oxide Emissions ◽  
Increase Production Efficiency ◽  
Plant Nitrogen ◽  
Rumen Microbes ◽  
Ruminant Meat ◽  
Choice Of Species ◽  
The Uk

Ruminant production in the UK is largely dependent on grasslands: approximately 52% of UK land is (improved) grassland and rough grazing. Herbage production from improved grassland can be over five times higher than that of indigenous swards (Davies et al., 1984) and these pastures account for the majority of ruminant (meat and milk) production from grassland the UK. They are also amenable to manipulation through the choice of species, variety and mixture of forages sown. Efficiency of production can be influenced at the levels of both the plant and the animal and by the nutrition and genetics of both. This paper examines strategies to increase production efficiency based upon optimising rumen fermentation linked to plant breeding approaches. In this way efficiency can be improved without recourse to diet manipulation with supplements, which is generally impractical at grazing. Although the rumen microbial population allows the ruminant animal to extract energy from otherwise unusable sources (i.e. fibres), the rumen is also the source of greatest inefficiencies in the use of nitrogen and energy. Degraded plant nitrogen that is not captured by the rumen microbes tends to be absorbed as ammonia, much of which is excreted as urea and contributes to ammonia and nitrous oxide emissions. Similarly, methane release from the rumen represents a waste of energy that could otherwise be used for production. Globally, these two processes result in ruminant livestock accounting for approximately a third of the methane emissions (Beauchemin et al., 2008) and half of the nitrous oxide emissions (de Klein and Eckard, 2008) from anthropogenic sources.

scholarly journals Management Information Needs of Fish Farmers in Egbeda Local Government Area of Oyo State

1970 ◽  
Vol 22 (1) ◽  
pp. 139-150
Author(s):  
Otobong Micheal Udo ◽  
L.A. Akinbile ◽  
Samuel Ikechukwu Chinkata
Keyword(s):  
Local Government ◽  
Information Needs ◽  
Production Efficiency ◽  
Management Practices ◽  
Water Quality Management ◽  
Fish Farming ◽  
Local Government Area ◽  
Management Information ◽  
Chi Square ◽  
Increase Production Efficiency

This study investigated the management information needs of fish farmers in Egbeda Local Government Area of Oyo State. Structured questionnaire was used to obtain data from one hundred and ten fish farmers randomly selected from four urban and seven rural wards of the area. Data obtained were analyzed using descriptive (frequency, percentage,) and inferential (chi-square and PPMC) statistics. Results show that respondents had average age of 43years, majority were male (64.8%), had primary school as highest educational level (52.4%), had 2-5 ponds, use earthen ponds (55.2%) and had 5-7 years fish farming experience. Information was most needed in hormone identification (x=2.83), fish marketing (x=2.80) and water quality management (x=2.71). Respondents indicated high cost of feed and poaching as a major challenge in fish farming. Respondents’ age (r = -0.233, p = 0.017); management practices (r = 0.209, p =0.032); number of ponds (r = -0.412, p = 0.000) were correlated with management information needs. The study therefore recommends the need for extension agencies to provide robust and timely information in the areas of need highlighted by the fish farmers in order to increase production efficiency and output of the fish farmers.Keywords: Information needs, Fish farmers, Aquaculture

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