204 Efficient editing of porcine parthenogenetic zygotes by electroporation of Cas9 ribonucleoprotein complexes

2019 ◽  
Vol 31 (1) ◽  
pp. 227
Author(s):  
F. L. Ongaratto ◽  
P. Rodriguez-Villamil ◽  
U. Ganbaatar ◽  
C. De Frutos ◽  
S. Solin ◽  
...  
Keyword(s):  
Large Scale ◽  
Gene Editing ◽  
The Other ◽  
Scale Production ◽  
Efficient System ◽  
Ribonucleoprotein Complexes ◽  
Group 4 ◽  
Group 2 ◽  
And Control ◽  
Group 1

Gene editing by microinjection is an efficient system to produce mutant livestock; however, microinjection is time-consuming and requires special skill, limiting its use for large-scale production of gene-edited animals. Therefore, the aim of this study was to develop a system to deliver guide (g)RNA/Cas9/ribonucleoprotein (RNP) by electroporation into parthenogenic porcine zygotes. For experiment 1, we delivered gRNA/Cas9 RNP (250ng μL−1 of each), targeting GATA4 using 2 electroporation conditions. Group 1 (n=130): 20V, 3ms, ×2 pulses, 1 repeat; group 2 (n=102): 20V, 1ms, ×2 pulses, 2 repeats; and Control (n=96): parthenogenic zygotes, no electroporation. For experiment 2, we delivered gRNA/Cas9 RNP (250ng μL−1 of each) targeting ROSA26 by electroporation with 4 conditions compared with delivery of RNP by microinjection: group 1 (n=17): 20V, 3ms, ×1 pulses, 1 repeat; group 2 (n=49): 20V, 3ms, ×3 pulses, 1 repeat; group 3 (n=64): 30V, 3ms, ×1 pulses, 1 repeat; group 4 (n=61): 30V, 3ms, ×3 pulses, 1 repeat; group 5 (n=120): zygotes microinjected with Cas9/ROSA26 sgRNA (25/25ng μL−1), and Control (n=76): parthenogenic zygotes, no electroporation. The electroporated zygotes were cultured in porcine zygote medium-3 (PZM-3) with controlled atmosphere, and development was evaluated on Day 2 (cleavage) and Day 7 (blastocyst rate). Gene editing was evaluated on embryos (blastocyst and morulas) by PCR and Sanger sequencing of amplicons including the RNP target site. Data were compared using chi-squared test, and differences were considered significant at P<0.05. Cleavage rates in experiment 1 were similar for the control (86/96; 89.5%), group 1 (94/102; 92.1%), and group 2 (119/130; 91.5%). Blastocyst rates were higher for the control (46/96; 47%) than for the other groups (P<0.01). However, for the treated groups, the blastocyst rates were similar, group 1 (19/102; 9.2%) and group 2 (12/130; 18.6%). Furthermore, the non-homologous end joining (NHEJ) efficiency was similar for groups 1 (14/18; 77.7%) and 2 (14/17; 82.3%). In experiment 2, the cleavage (53/76; 69%) and blastocyst rates (30/76; 39%) were significantly higher for the control than for the treated groups (P<0.01). Among the groups, the lower cleavage and blastocyst rates were for group 4 (20/61; 32.7% and 3/61; 4.9%, respectively) compared with the other electroporation and microinjection groups (P<0.03). However, NHEJ efficiency was higher for electroporation groups 2 (6/8; 75%), 3 (17/17; 100%), and 4 (2/2; 100%) compared with microinjection (2/15; 13%). In conclusion, electroporation of Cas9/RNP is an efficient alternative to microinjection for gene editing in porcine zygotes.

scholarly journals Treatment of ovarian cysts in sows – a field trial

2008 ◽  
Vol 52 (No. 9) ◽  
pp. 413-418 ◽  
Author(s):  
S. Cech ◽  
R. Dolezel
Keyword(s):  
The Other ◽  
Ovarian Cysts ◽  
Control Group ◽  
Initial Examination ◽  
Interval Group ◽  
Group 4 ◽  
Group 3 ◽  
Group 5 ◽  
Group 2 ◽  
Group 1

Different procedures of treatment of large follicular ovarian cysts in 177 sows using GnRH, hCG and PGF2α are evaluated in this study. Ovarian cysts were diagnosed by transcutaneous ultrasonography, which was a part of routine pregnancy diagnosis. No treatment was performed in the control group (Group 1, <i>n</i> = 29); the method of treatment used in the other groups immediately after the diagnosis was intramuscular administration of lecirelin in doses 50 µg (Group 2, <i>n</i> = 28), 100 µg (Group 3, <i>n</i> = 27) and 200 µg divided into 2 equal doses administered at a 12-hour interval (Group 4, <i>n<i> = 25) and of hCG in doses 1 500 IU (Group 5, <i>n</i> = 23), 3 000 IU (Group 6, <i>n</i> = 21), and 250 µg of cloprostenol (Group 7, <i>n</i> = 24). Insemination rate (IR), conception rate (CR) in inseminated sows, pregnancy rate (PR = recovery rate), treatment-insemination interval (TII) and treatment-pregnancy interval (TPI) within 42 days after the initial examination were evaluated. In addition PR in groups of sows divided according to parity (1–3, 4–6 and ≥ 7) were also evaluated. IR and PR were higher in Group 4 (84.0% and 44.0%) and lower in Group 1 (17.2% and 6.9%) in comparison with the other groups (<i>P</i> < 0.001 and <i>P</i> < 0.05). CR, TII and TPI did not differ between the experimental groups. PR were similar in sows with different parity. The study proved a positive response in sows with large follicular ovarian cysts to the treatment consisting of 2 administrations of 100 µg GnRH at a 12-hour interval.

scholarly journals P0314TREATMENT AND OUTCOMES OF IDIOPATHIC MEMBRANOUS NEPHROPATHY IN ELDERLY PATIENTS

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Nessrine Breik ◽  
Mouna Jerbi ◽  
Raja Aoudia ◽  
Soumaya Chargui ◽  
Hanen Guaied ◽  
...  
Keyword(s):  
Nephrotic Syndrome ◽  
Side Effects ◽  
Immunosuppressive Therapy ◽  
Older Patients ◽  
Symptomatic Treatment ◽  
The Elderly ◽  
The Other ◽  
Group 4 ◽  
Group 2 ◽  
Group 1

Abstract Background and Aims The increase in life expectancy has led to increasing numbers of elderly patients in all medical disciplines, particularly in nephrology. Idiopathic membranous nephropathy (IMN) is common in the elderly and can lead to significant morbidity and mortality because of the complication of nephrotic syndrome (NS) and immunosuppressive therapy. The aim of this study is to analyze the diagnostic, therapeutic and evolutionary approach of IMN in elderly patiets. Method We conducted a retrospective descriptive study in the nephrology department at Charles Nicolle hospital over a period of 44 years. All older patients (≥65 years) with histologically proven MN were included in this study. Data collected included demographic, clinical and biological parameters in each patient. Data were entered and analyzed using SPSS software. Results Twenty-eight patients were collected. The mean age was 67.03 years (65-78 years) with a male predominance (sex ratio: 2.3) and low socio-economic level in 82.5% of cases. Sixteen patients were smokers (57.14%), 5 ethyl patients (17.8%), diabetes was present in 3 patients (10.7%) and hypertension in 11 patients (39.28%). Two cases of neoplasm were present, namely one case of prostatic adenocarcinoma and one case of gallbladder adenocarcinoma, all were diagnosed and treated along one year and ten years respectively, before the diagnosis of MN. The circumstances of discovery were dominated by oedema in 27 cases (86.27%), hypertension in 11 cases (39.28%) and elevated creatinine level in 9 cases (32.14%). Deep venous thrombosis was the circumstance of discovery in one case. At the time of diagnosis, the clinico-biological picture was dominated by high systolic blood pressure in 21 cases (75%), anasarca in 7 cases (25%), proteinuria in all cases and hematuria in 20 cases (71.14%). Biology revealed nephrotic syndrome (NS) in all cases, hypercholesterolemia in 23 cases (82.14%), high serum creatinine in 14 cases (50%) with an average creatinine level of 127,95 µmol/l, anemia in 17 cases (60.7%) and anti-neutrophil cytoplasmic antibodies were positive in one case. MN was confirmed by a kidney biopsy in all cases. Symptomatic treatment was indicated in all patients. Immunosuppressive therapy was started early in 12 patients (40%) because of the severe NS and the deterioration of renal function. Eight patients (26.6%) received corticosteroids alone (group 1), three patients received corticosteroid with mycofenolate mofetil (group 2) and one patient received corticosteroid with ciclosporin (group3). Fourteen patients received only symptomatic treatment (group 4). We noted partial remission in group 3, however complete remission in 37.5%, 14.28% and 12.5% from respectively group 1, group 4 and group 2. The side effects of immunosuppressive therapy were infectious complication in 8 patients (28.5%), osteoporosis in 3 patients (10.7%) and Steroid diabetes in one case. The side effects of NS were thrombotic complication in 5 patients (17.85%). Two patients had presented coronary syndrome and two other patients had presented hepatitis C. Two cases of death were noted whose etiology was pulmonary infection in one patient and undeterminate etiology in the other case. Conclusion Treatment of IMN in older patients has unique challenges, reducing the need for renal replacement therapy. Most studies report that the elderly respond to therapy with rates comparable with younger patients but in the other hand elderly are more susceptible to side effects of immunosuppressive therapy.

scholarly journals Pregnancy rates in criollo breed mares after ultrasound-guided follicular aspiration

2014 ◽  
Vol 15 (3) ◽  
pp. 339-343
Author(s):  
Vivian Campos Laia Franco ◽  
Fabrício Desconsi Mozzaquatro ◽  
Rodrigo Arruda de Oliveira ◽  
Flávio Desessards De La Corte ◽  
Mara Iolanda Batistella Rubin ◽  
...  
Keyword(s):  
Pregnancy Rates ◽  
Control Group ◽  
Ultrasound Guided ◽  
Group 4 ◽  
Follicular Aspiration ◽  
Future Fertility ◽  
Group 3 ◽  
Group 2 ◽  
And Control ◽  
Group 1

There are few studies about transvaginal ultrasound-guided follicle aspiration in equine medicine regarding potential complications to future fertility of aspirated mares. In order to evaluate the effect of follicular aspiration on subsequent fertility in mares, two experiments were conducted. In Experiment I, fifteen Criollo mares were allocated to one of three groups according to the diameter of the aspirated follicle during estrus: 25-29mm (n=4; Group 1); 30-34mm (n=6; Group 2); > 35mm (n=5; Group 3) and control group (n=15; Group 4). In Experiment II, the follicular aspiration was attempted in twenty-five mares during diestrous, when at least four follicles (> 5mm) were seen in the transrectal ultrasonography of both ovaries. All visible follicles, between 4 and 8 mm, were aspirated. Thirty-one mares served as control. In Experiments I and II, the pregnancy rates in the following cycle after aspiration were 75.0% (Group 1), 83.3% (Group 2), 60.0% (Group 3), and 73.3% (Group 4 - Control); and 76.0% in the aspirated diestrous group and 77.4% in the control group (non aspirated), respectively. On both experiments, pregnancy rates were similar (P>0.05) in treated and control mares. The results of this study show that the conception rates of the first estrus period following follicular aspiration are not affected by the procedure.

Influence of Cavity Preparation, Light-curing Units, and Composite Filling on Intrapulpal Temperature Increase in an In Vitro Tooth Model

2014 ◽  
Vol 39 (5) ◽  
pp. E195-E205 ◽  
Author(s):  
SH Choi ◽  
JF Roulet ◽  
SD Heintze ◽  
SH Park
Keyword(s):  
Resin Composite ◽  
The Other ◽  
Temperature Measurements ◽  
Group 4 ◽  
Light Curing ◽  
Group 3 ◽  
Group 2 ◽  
Power Densities ◽  
Group 1

SUMMARY This study examined the effect of both the tooth substance and restorative filling materials on the increase in pulp chamber temperature when using light-curing units with different power densities. The tip of a temperature sensor was positioned on the pulpal dentinal wall of the buccal side of a maxillary premolar. Metal tubes were inserted in the palatal and buccal root of the tooth, one for water inflow and the other for water outflow. Polyethylene tubes were connected from the metal tubes to a pump to control the flow rate. For the unprepared tooth group (group 1), the tooth was light-cured from the buccal side using two light-curing units (three curing modes): the VIP Junior (QTH, BISCO, Schaumburg, IL, USA) and the Bluephase LED light-curing units (two modes: LEDlow and LEDhigh; Ivoclar Vivadent, Schaan, Liechtenstein). The power densities of each light-curing unit for the LEDlow, QTH, and LEDhigh modes were 785 mW/cm2, 891 mW/cm2, and 1447 mW/cm2, respectively. All light-curing units were activated for 60 seconds. For the prepared tooth group (group 2), a Class V cavity, 4.0 mm in width by 4.0 mm in height by 1.8 mm in depth in size, was prepared on the buccal surface of the same tooth for the temperature measurement. The light-curing and temperature measurements were performed using the same methods used in group 1. The cavity prepared in group 2 was filled with a resin composite (Tetric N Ceram A3 shade, Ivoclar Vivadent) (group 3) or a flowable composite (Tetric N Flow with A3 shade, Ivoclar Vivadent) (group 4). The light-curing and temperature measurements were performed for these groups using the same methods used for the other groups. The highest intrapulpal temperature (TMAX) was measured, and a comparison was conducted between the groups using two-way analysis of variance with a post hoc Tukey test at the 95% confidence level. The TMAX values were as follows: 38.4°C (group 1), 39.0°C (group 2), 39.8°C (group 3), and 40.3°C (group 4) for the LEDlow mode. For the QTH mode, the TMAX values were 40.1°C (group 1), 40.4°C (group 2), 40.9°C (group 3), and 41.4°C (group 4). For the LEDhigh mode, the TMAX values were 43.3°C (group 1), 44.5°C (group 2), 44.7°C (group 3), and 45.3°C (group 4). The statistical analysis revealed the following: the TMAX values were arranged by mode in the following manner: LEDlow &lt; QTH &lt; LEDhigh (p&lt;0.05) and group 1 &lt; group 2 ≤ group 3 ≤ group 4 (p&lt;0.05).

scholarly journals An analysis of pH, pO2 and pCO2 in the peritoneal fluid of dogs with ascites of various etiologies

2016 ◽  
Vol 19 (1) ◽  
pp. 141-145 ◽  
Author(s):  
K. Glińska-Suchocka ◽  
P. Sławuta ◽  
M. Jankowski ◽  
K. Kubiak ◽  
J. Spużak ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Peritoneal Fluid ◽  
Proteolytic Enzymes ◽  
Abdominal Cavity ◽  
The Other ◽  
Bacterial Peritonitis ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Abstract The aim of the study was to assess pH, pO2 and pCO2 in peritoneal fluid. The study was conducted on a group of 22 dogs with symptoms of ascites. Group 1 consisted of 4 dogs with adenocarcinoma, group 2 - of 6 dogs with glomerulonephritis, group 3 of 8 dogs with hepatic cirrhosis and group 4 of 4 dogs with bacterial peritonitis. An abdominal cavity puncture was performed in all dogs and the fluid was drawn into a heparinized syringe in order to assess pH, pO2 and pCO2 . The analysis of pH in the peritoneal fluid revealed statistically significant differences between group 4 and groups 1 (p=0.01), 2 (p=0.01), and 3 (p=0.01). The lowest pH value compared to the other studied groups was recorded in group 4. In group 4, the pO2 was the lowest compared to the other groups (group 1 p=0.01, group 2 p=0.01, group 3 p=0.01). The value of pCO2 was the highest in group 4 compared to groups 1, 2, and 3. The study found statistically significant differences in pH, pCO2 and pCO2 between group 4 (the group of dogs with bacterial peritonitis) and the other groups of dogs. This was probably linked to the pathogenesis of peritonitis. As a result of an inflammatory reaction within the peritoneal cavity, there is an increase in fibrin accumulations leading to a decreased oxygen supply causing the oxidative glucose metabolism to change into a non-oxidative glucose metabolism. This, in turn, causes a decrease in pH, acidosis, and a low oxidoreduction potential. It also impairs phagocytosis and activates proteolytic enzymes which create ideal conditions for the growth of anaerobic bacteria. The obtained results indicate that the pH, pO2 and pCO2 may be used to differentiate bacterial peritonitis from ascites of other etiologies.

Comparative Data about Estrus Induction and Pregnancy Rate on Lacaune Ewes in Non-breeding Season after Melatonin Implants and Intravaginal Progestagen

2021 ◽  
Author(s):  
D. Berean ◽  
O. Ergene ◽  
A. Blaga-Petrean ◽  
I. Bogdan ◽  
S. Ciupe ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Fixed Time ◽  
Control Group ◽  
Group 4 ◽  
Induction Rate ◽  
Group 3 ◽  
Group 2 ◽  
And Control ◽  
Control Group Group ◽  
Group 1

Backgroung: Seasonal anestrus in ewes reduces reproductive efficiency and hinders productivity. Exogenous hormones can be used to facilitate the induction and synchronization of estrus in the anestrus period. The aim of this study was to evaluate the pregnancy rate in ewes after the treatment with different hormones and fixed time artificial insemination. Methods: The ewes were randomly divided in 4 (n=4) groups: Melatonin group (Group 1), Melatonin and Medroxyprogesterone group (Group 2), Medroxyprogesterone group (Group 3) and control group (Group 4). Pregnancy rate, estrus induction rate and litter size were determined and compared. Result: The estrus induction rate in our study was 100% for the first 3 groups and 6.25% for the 4th group. The pregnancy rate was between 4.16% (group 4) and 95.83% (group 2). For the others groups the values were 89.58% for the first group and 91.67%, for the 3rd group. The prolificacy rate was between 1 (control group) and 1.71 (2nd group), with intermediate values for the other 2 groups: 1.21 for 1st group and 1.27 for the 3rd group.

scholarly journals EFEITO DOS PROCEDIMENTOS DE ACABAMENTO E POLIMENTO NA RUGOSIDADE SUPERFICIAL DA CERÂMICA NORITAKE®

2016 ◽  
Vol 7 (2) ◽  
Author(s):  
Mario Cezar Oliveira ◽  
Alex Correia Vieira ◽  
Adriana Castro Andrade ◽  
Indira Almeida Oliveira
Keyword(s):  
Surface Roughness ◽  
The Other ◽  
Control Group ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Diamond Paste ◽  
Group 1

The aim of this study was to evaluate the surface roughness of Noritake® ceramic after different methods of finishing and polishing. 40 specimens of Noritake® ceramic were randomly divided into 4 groups (n=10) and subjected to different methods of finishing and polishing: Group 1 (control) - glaze; Group 2 - abrasive rubbers Astropol (Ivoclar Vivadent), felt disc and diamond paste; Group 3 - abrasive rubbers (Edenta), felt disc and diamond paste; Group 4 - Sof-Lex discs (3M ESPE), felt disc and diamond paste. Subsequently, the surface roughness was measured with rugosimeter (Mitutoyo SJ 301). The control group (G1) had the lowest surface roughness (0.68 μm) that was different statistically to the other groups (p0.05). The different methods of finishing and polishing were not able to promote a surface as smooth as the application of glaze in the ceramics tested.

The Penetration of Ofloxacin into Human Aqueous Humor Given by Various Routes

1998 ◽  
Vol 8 (1) ◽  
pp. 33-36 ◽  
Author(s):  
S. Hanioglu-Kargi ◽  
N. Basci ◽  
H. Soysal ◽  
A. Bozkurt ◽  
E. Gürsel ◽  
...  
Keyword(s):  
Aqueous Humor ◽  
Bacterial Infections ◽  
High Performance ◽  
Cataract Extraction ◽  
The Other ◽  
Group 4 ◽  
Significant Difference ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

This study was designed to measure the concentration of ofloxacin in aqueous humor after topical, oral and intravenous administration in 50 patients undergoing cataract extraction. In Group 1, ofloxacin 0.3% eyedrops were topically instilled ten times and the aqueous humor concentration was 2.73 ± 0.88 μg/ml. In Group 2, ofloxacin 0.3% eyedrops were topically instilled six times and the aqueous humor concentration was 0.84 ± 0.61 μg/ml. Aqueous humor concentration 12 hours after 200 mg oral dose in Group 3, was 0.38 ± 0.12 μg/ml. In Group 4, patients were given ofloxacin as a single intravenous 200 mg dose and the aqueous humor concentration 2 hours after the end of infusion was 0.45 ± 0.11 μg/ml. Concentrations were determined by high performance liquid chromatography (HPLC) with fluorescence detection. There was a significant difference between Group 1 and the other groups, but not between Group 2 and Groups 3, 4. It was concluded that ofloxacin penetrates the corneal and the blood-aqueous barriers and can achieve good aqueous levels when given topically and systematically. Ofloxacin can be applied topically for external bacterial infections such as conjunctivitis and keratitis. Systematically administered ofloxacin reached higher levels than the MIC for some bacteria which cause endophthalmitis.

Transfer Effects of Learning at Specific Speeds on Performance over a Range of Speeds

1980 ◽  
Vol 50 (1) ◽  
pp. 83-89 ◽  
Author(s):  
Donald Siegel ◽  
Christine Davis
Keyword(s):  
Training Group ◽  
The Other ◽  
Transfer Effects ◽  
Group 4 ◽  
Pursuit Rotor ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

The effect of learning a novel skill at specific speeds on performance over a range of speeds was examined on the pursuit rotor. Three groups of subjects were given three days of training: Group 1 at 30 rpm, Group 2 at 60 rpm, and Group 3 at 30–45.60 rpm. Group 4, a control, practiced on a pegboard task during this period. On Days 4 and 5, all four groups were tested for transfer at 30, 45, and 60 rpm. For the most part, Group 3 appeared to perform equal to or better across the range of speeds than any of the other groups.

In-vivo evaluation of the effect of cyanoacrylate on prosthetic vascular graft infection – does cyanoacrylate increase the severity of infection?

VASA ◽  
2020 ◽  
Vol 49 (4) ◽  
pp. 281-284
Author(s):  
Atıf Yolgosteren ◽  
Gencehan Kumtepe ◽  
Melda Payaslioglu ◽  
Cuneyt Ozakin
Keyword(s):  
Vascular Graft ◽  
Graft Infection ◽  
Vascular Graft Infection ◽  
Group 4 ◽  
Significant Difference ◽  
Group 3 ◽  
Group 2 ◽  
Prosthetic Vascular Graft Infection ◽  
Group 1

Summary. Background: Prosthetic vascular graft infection (PVGI) is a complication with high mortality. Cyanoacrylate (CA) is an adhesive which has been used in a number of surgical procedures. In this in-vivo study, we aimed to evaluate the relationship between PVGI and CA. Materials and methods: Thirty-two rats were equally divided into four groups. Pouch was formed on back of rats until deep fascia. In group 1, vascular graft with polyethyleneterephthalate (PET) was placed into pouch. In group 2, MRSA strain with a density of 1 ml 0.5 MacFarland was injected into pouch. In group 3, 1 cm 2 vascular graft with PET piece was placed into pouch and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. In group 4, 1 cm 2 vascular graft with PET piece impregnated with N-butyl cyanoacrylate-based adhesive was placed and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. All rats were scarified in 96th hour, culture samples were taken where intervention was performed and were evaluated microbiologically. Bacteria reproducing in each group were numerically evaluated based on colony-forming unit (CFU/ml) and compared by taking their average. Results: MRSA reproduction of 0 CFU/ml in group 1, of 1410 CFU/ml in group 2, of 180 200 CFU/ml in group 3 and of 625 300 CFU/ml in group 4 was present. A statistically significant difference was present between group 1 and group 4 (p < 0.01), between group 2 and group 4 (p < 0.01), between group 3 and group 4 (p < 0.05). In terms of reproduction, no statistically significant difference was found in group 1, group 2, group 3 in themselves. Conclusions: We observed that the rate of infection increased in the cyanoacyrylate group where cyanoacrylate was used. We think that surgeon should be more careful in using CA in vascular surgery.

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