scholarly journals Intrafollicular oocyte transfer in cattle – a technical report

2020 ◽  
Vol 89 (1) ◽  
pp. 11-17
Author(s):  
Michaela Andrlikova ◽  
Vladislav Bína ◽  
Vojtech Kos ◽  
Miloslava Lopatářová ◽  
Beata Markova ◽  
...  
Keyword(s):  
Preovulatory Follicle ◽  
Recovery Rates ◽  
Total Recovery ◽  
Embryo Recovery ◽  
Technical Report ◽  
New Instrument ◽  
Preovulatory Follicles ◽  
Phosphate Buffered Saline

The aim of the study was to develop and evaluate the functionality of a new equipment for intrafollicular oocyte transfer (IFOT) in dairy cattle. The new system for IFOT is composed of the applicator, the aspirator, and the injector. After aspiration of oocytes, the IFOT set is inserted into the working tube of the ultrasound transducer holder, the content of the applicator can be injected into the preovulatory follicle via transvaginal ultrasonography by one operator. The function of instruments used for IFOT was firstly verified in laboratory conditions. Slaughterhouse oocytes filled into the instruments were injected into Petri dishes. The highest recovery rates in vitro (97.5%) were achieved when the applicator was stored with the needle in a downward position. Synchronized Holstein heifers were used for in vivo test. Intrafollicular injection of saline (n = 9) was performed to find whether ovulation is affected by the injection. Then IFOTs of phosphate buffered saline with 20 oocytes (n = 21) were performed into the preovulatory follicles followed by 7-day-old embryo collection. Total ovulation rates were 86.7% (26/30). Total recovery rates (oocytes + embryos) were 23.1%, embryo recovery rates were 10.1%. The new instrument allows for the loading of oocytes and easy transportation to recipients, and also allows IFOT to be performed by one person in field conditions. The method does, however, need further investigation.

Improved in ovulo embryo culture for stenospermocarpic grapes (Vitis vinifera L.)

2003 ◽  
Vol 54 (9) ◽  
pp. 869 ◽  
Author(s):  
S. M. Liu ◽  
S. R. Sykes ◽  
P. R. Clingeleffer
Keyword(s):  
Embryo Rescue ◽  
Vitis Vinifera L ◽  
Embryo Survival ◽  
Recovery Rates ◽  
Continuous Growth ◽  
Embryo Recovery ◽  
Plantlet Formation ◽  
Seedless Grape

In ovulo embryo rescue techniques have been used to recover new hybrids from seedless × seedless grape crosses. This study was conducted to increase efficiency by investigating effects of genotype, medium, and ovule removal age on ovule elongation, embryo recovery, growth, and plantlet formation. Ovules from self-pollinated berries of seedless varieties Sunmuscat, Merbein Seedless, and Marroo Seedless were cultured at 30, 43, 60, and 70 days after flowering (DAF) in a range of media, some of which were supplemented with gibberellic acid (GA3) and indole-3-acetic acid (IAA). The effect of activated charcoal (AC) in media on rescued embryos was also investigated. Ovules exhibited continuous growth in vivo and in vitro. The most vigorous growth was observed for ovules cultured at 30 and 43 DAF, but more embryos were recovered from ovules cultured at 60 and 70 DAF. Ovule growth and embryo production in vitro were improved in Bouquet and Davis (BD) and Nitsch and Nitsch (NN) media. Supplementation with GA3 increased embryo recovery rates. Highest embryo recovery rates were 18.1%, 9.6%, and 12.2% for Sunmuscat, Merbein Seedless, and Marroo Seedless, respectively, when ovules were excised and cultured at 60 or 70 DAF in either BD or NN media. In vitro embryo survival and plantlet formation were higher for torpedo-shaped embryos, and improved greatly in 6-benzyladenine (BA)-supplemented woody plant (WP) medium containing 0.3% AC. Embryo recovery was improved by excising and culturing ovules at 60 DAF in BD or NN media and then by transferring embryos to WP medium supplemented with BA and AC.

Temperature gradients in vivo influence maturing male and female gametes in mammals: evidence from the cow

2017 ◽  
Vol 29 (12) ◽  
pp. 2301 ◽  
Author(s):  
R. H. F. Hunter ◽  
F. López-Gatius ◽  
O. López-Albors
Keyword(s):  
Follicular Fluid ◽  
Open Surgery ◽  
Temperature Gradients ◽  
Fluid Temperature ◽  
Preovulatory Follicle ◽  
Male And Female ◽  
Direct Measurements ◽  
Final Maturation ◽  
Preovulatory Follicles

Since 1980 several reports have indicated that temperatures vary between preovulatory follicles and other ovarian tissues in rabbit, cow, pig and human. However, these observations did not achieve prominence; they were regarded as artefacts due to the use of anaesthetics and open surgery (laparotomy). Recently, without resorting to anaesthesia or surgery, direct measurements of temperature in preovulatory follicles have been performed in the cow by means of a thermistor probe introduced into the antrum under ultrasonic guidance. Such follicles revealed a mean antral (follicular fluid) temperature 0.74°C and 1.54°C cooler than uterine surface and rectal temperatures respectively in ovulating cows, whereas no such temperature differences were detected in non-ovulating cows. Cows are predominantly monovular and preovulatory follicles attain a diameter of 15–22 mm or more. These features and the timescale of response to the preovulatory gonadotrophin surge make them a valuable model for the human preovulatory follicle. Temperature gradients are interpreted primarily in a context of final maturation of gametes immediately before the onset of fertilisation. Preovulatory follicular temperature in women could be assessed by a comparable approach and might become a valuable selection guide for oocyte viability.

scholarly journals Gonadotropin-Induced Expression of Messenger Ribonucleic Acid for Cyclooxygenase-2 and Production of Prostaglandins E and F2α in Bovine Preovulatory Follicles Are Regulated by the Progesterone Receptor

Endocrinology ◽  
2006 ◽  
Vol 147 (10) ◽  
pp. 4713-4722 ◽  
Author(s):  
P. J. Bridges ◽  
C. M. Komar ◽  
J. E. Fortune
Keyword(s):  
Granulosa Cells ◽  
Medroxyprogesterone Acetate ◽  
Cyclooxygenase 2 ◽  
Induced Expression ◽  
Messenger Ribonucleic Acid ◽  
Preovulatory Follicles ◽  
Gonadotropin Surge ◽  
Induced Changes

Follicular production of prostaglandins (PGs) is essential for ovulation, but the factors mediating gonadotropin-induced secretion of PGE and PGF2α remain largely unknown. We tested the hypothesis that gonadotropin-induced changes in progesterone and its receptor (PR) mediate the increase in periovulatory PGs. Heifers were treated with PGF2α and GnRH to induce luteolysis and the LH/FSH surge (ovulation occurs ∼30 h after GnRH). Because there are two increases in intrafollicular progesterone/PR mRNA during the bovine periovulatory period, we first examined the temporal pattern of PG production by follicles collected at 0, 3.5, 6, 12, 18, and 24 h after GnRH. Although PGs did not increase in the follicular fluid until 24 h after GnRH, acute secretion of PGs by follicle wall (theca + granulosa cells) was initiated by 18 h and had increased manyfold by 24 h after GnRH. In vitro, FSH and LH induced dramatic transient increases in PG production by follicle wall and granulosa, but not theca, cells isolated from preovulatory follicles (0 h after GnRH). PG accumulation peaked on d 2 of culture, mimicking the secretion pattern after a gonadotropin surge in vivo. In cultures of follicle wall and granulosa cells, the PR antagonist mifepristone (MIFE, 1 μm) inhibited LH-induced PG secretion and the progestin medroxyprogesterone acetate (1 or 10 μm), but not the glucocorticoid dexamethasone (1 or 10 μm), overcame the effect of MIFE on PGs. Semiquantitative RT-PCR revealed that MIFE inhibited LH-induced expression of cyclooxygenase-2 mRNA in granulosa cells in vitro. Again, treatment with medroxyprogesterone acetate overcame the effect of MIFE. Together these results provide strong evidence that periovulatory increases in cyclooxygenase-2 mRNA, PGE, and PGF2α are mediated by gonadotropin-induced increases in progesterone/PR, indicating that in some species there is an important functional relationship between these pathways in the ovulatory cascade.

scholarly journals Drug Release Kinetics of Electrospun PHB Meshes

Materials ◽  
2019 ◽  
Vol 12 (12) ◽  
pp. 1924 ◽  
Author(s):  
Vojtech Kundrat ◽  
Nicole Cernekova ◽  
Adriana Kovalcik ◽  
Vojtech Enev ◽  
Ivana Marova
Keyword(s):  
Release Kinetics ◽  
Entrapment Efficiency ◽  
Toxic Substances ◽  
Antimicrobial Efficiency ◽  
Vis Spectroscopy ◽  
Model Drug ◽  
Phosphate Buffered Saline ◽  
Kinetics Of

Microbial poly(3-hydroxybutyrate) (PHB) has several advantages including its biocompatibility and ability to degrade in vivo and in vitro without toxic substances. This paper investigates the feasibility of electrospun PHB meshes serving as drug delivery systems. The morphology of the electrospun samples was modified by varying the concentration of PHB in solution and the solvent composition. Scanning electron microscopy of the electrospun PHB scaffolds revealed the formation of different morphologies including porous, filamentous/beaded and fiber structures. Levofloxacin was used as the model drug for incorporation into PHB electrospun meshes. The entrapment efficiency was found to be dependent on the viscosity of the PHB solution used for electrospinning and ranged from 14.4–81.8%. The incorporation of levofloxacin in electrospun meshes was confirmed by Fourier-transform infrared spectroscopy and UV-VIS spectroscopy. The effect of the morphology of the electrospun meshes on the levofloxacin release profile was screened in vitro in phosphate-buffered saline solution. Depending upon the morphology, the electrospun meshes released about 14–20% of levofloxacin during the first 24 h. The percentage of drug released after 13 days increased up to 32.4% and was similar for all tested morphologies. The antimicrobial efficiency of all tested samples independent of the morphology, was confirmed by agar diffusion testing.

Contribution of LFA-1 and Mac-1 to CD18-dependent neutrophil emigration in a neonatal rabbit model

1996 ◽  
Vol 80 (6) ◽  
pp. 1984-1992 ◽  
Author(s):  
J. M. Graf ◽  
C. W. Smith ◽  
M. M. Mariscalco
Keyword(s):  
Monoclonal Antibody ◽  
Polymorphonuclear Leukocytes ◽  
Rabbit Model ◽  
Transendothelial Migration ◽  
Peritoneal Exudate ◽  
Adult Rabbit ◽  
Primary Mechanism ◽  
Phosphate Buffered Saline

Human neonatal polymorphonuclear leukocytes (PMNs) exhibit decreased mobility, adherence, and transendothelial migration in vitro compared with adult PMNs. These deficits, in part, are due to functional and quantitative defects in neonatal Mac-1 (CD11b/CD18), whereas LFA-1 (CD11a/CD18) function is similar to that found in adults (D.C.Anderson, O.Abbassi, T.K.Kishimoto, J.M.Koenig, L.V.McIntire, and C.W.Smith, J.Immunol. 146: 3372-3379, 1991; C. W. Smith, T. K. Kishimoto, O. Abbassi, B.J.Hughes, R.Rothlein, L.V.McIntire, E.Butcher, and D.C. Anderson, J. Clin. Invest. 87: 609-618, 1991). We tested the hypothesis that the primary mechanism for the neonatal PMN CD18-dependent emigration in vivo is due to LFA-1. Neutrophils from 1-day-old rabbit pups had 32 and 60% of adult rabbit levels of CD11a and CD11b, respectively. Rabbit pups or adult rabbits received the monoclonal antibody (MAb) R7.1 (anti-CD11a) or R15.7 (anti-CD18) or the vehicle phosphate-buffered saline (PBS) before the instillation of intraperitoneal thioglycollate. Six hours later peritoneal exudate was collected. The administration of MAbs R7.1 and R15.7 in adult animals resulted in 60 and 83% inhibition of leukocyte emigration, respectively, compared with PBS-treated animals (P < 0.01). In neonatal animals, R7.1 and R15.7 inhibited leukocyte peritoneal accumulation to the same extent (50 and 60%, respectively) compared with PBS-treated animals (P < 0.01). Adult animals were also treated with the anti-CD11b MAb 198. MAb 198 decreased emigration by 25%, although this was not significant compared with PBS-treated animals. We conclude that although neonatal animals have significantly less neutrophil CD11a, the diminished levels did not contribute to a reduced ability to emigrate to the peritoneum and, like adult animals, neonatal animals primarily utilize LFA-1 for accumulation in this model. The contribution of Mac-1 to neonatal leukocyte emigration remains uncertain.

Effect of gonadotrophins on progesterone secretion by cultured granulosa cells obtained from human preovulatory follicles

1985 ◽  
Vol 110 (3) ◽  
pp. 401-407 ◽  
Author(s):  
T. Hillensjö ◽  
A. Sjögren ◽  
B. Strander ◽  
L. Nilsson ◽  
M. Wikland ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Maximal Effect ◽  
Tubal Infertility ◽  
Vitro Fertilization ◽  
Follicular Maturation ◽  
Progesterone Secretion ◽  
Preovulatory Follicles ◽  
Ultrasound Guided Puncture

Abstract. Granulosa cells were obtained from human preovulatory follicles in 31 women undergoing in vitro fertilization and embryo transfer due to tubal infertility. Follicular maturation was stimulated and synchronized by treatment with Clomiphene or human menopausal gonadotrophin (hMG), or both, plus human chorionic gonadotrophin (hCG). Follicles were aspirated by ultrasound guided puncture approximately 34–36 h after the hCG injection. The granulosa cells were washed and suspended in modified medium 199 containing 10% foetal bovine serum and cultured as monolayers for 6–8 days in the absence and presence of hormones and reactants. Progesterone formation was analyzed by RIA. In general, the cells underwent morphological luteinization and secreted high amount of progesterone. Under basal conditions the secretion of progesterone was highest during the first 2 days in culture and then gradually declined. Progesterone secretion was stimulated by human LH, hCG and the adenylate cyclase stimulator forskolin, with a maximal effect between days 2–6. The β-adrenergic agonist isoproteronol in preliminary experiments potentiated the stimulatory effect of hCG but had no own stimulatory effect. No clear differences in progesterone secretion or responsiveness to in vitro stimulation relating to the various in vivo stimulation protocols were found.

Effects of the antiandrogen hydroxyflutamide on progesterone secretion by preovulatory rat follicles in vivo and in vitro

1991 ◽  
Vol 69 (9) ◽  
pp. 1288-1293 ◽  
Author(s):  
Yallampalli Chandrasekhar ◽  
David T. Armstrong
Keyword(s):  
Luteinizing Hormone ◽  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
Serum Progesterone ◽  
Antagonistic Action ◽  
Lh Surge ◽  
Progesterone Secretion ◽  
Preovulatory Follicles

Serum and ovarian progesterone levels and in vitro production of progesterone by preovulatory follicles were measured on proestrus in pregnant mare's serum gonadotropin (PMSG) primed immature rats in which the luteinizing hormone (LH) surge and ovulation were blocked by administration of the antiandrogen hydroxyflutamide. Serum progesterone levels observed at 12:00 on proestrus were significantly elevated, twofold above those observed in vehicle-treated controls, by in vivo administration of 5 mg hydroxyflutamide 4 h earlier. In control rats, proestrous progesterone did not increase until 16:00, in parallel with rising LH levels of the LH surge. No LH surge occurred in the hydroxyflutamide-treated rats, ovulation was blocked, and serum progesterone declined throughout the afternoon of proestrus, from the elevated levels present at 12:00. Administration of human chorionic gonadotropin (hCG) at 11:00 advanced the elevation of serum progesterone by 2 h in vehicle-treated controls and prevented the decline in progesterone levels in hydroxyfiutamide-treated rats. The patterns of change in ovarian tissue concentrations with time and treatment were essentially similar to those observed for serum progesterone. In in vitro experiments, progesterone secretion during 24 h culture of preovulatory follicles obtained on PMSG-induced proestrus was significantly increased, sixfold, by addition to the culture media of 370 μM but not of 37 μM hydroxyflutamide. Testosterone (50 nM) and hCG (20 mIU/mL) caused 26- and 14-fold increases, respectively, in progesterone secretion by cultured follicles. Hydroxyflutamide significantly reduced the stimulatory effect of testosterone but not of hCG on progesterone secretion in vitro. These results suggest that the antiandrogen hydroxyflutamide stimulates progesterone secretion, both in vivo and in vitro, through an initial androgen-agonistic action, before its antagonistic action is expressed. Its androgen-antagonistic action is responsible for its ability to inhibit testosterone-induced progesterone secretion in vitro. Its failure to reduce hCG-stimulated progesterone secretion in vivo and in vitro indicates that the latter stimulation is exerted independently of, and not as a consequence of, androgen action. The decrease in serum progesterone levels on the afternoon of proestrus therefore appears to be a consequence rather than a cause of the absence of an LH surge in the hydroxyflutamide-treated rats. It is concluded that the inhibitory effect of hydroxyflutamide on the preovulatory LH surge and ovulation is due not to inhibition of progesterone secretion at the ovarian level but most likely to neuroendocrine site(s) of action of the inhibitor.Key words: antiandrogen, hydroxyflutamide, progesterone, luteinizing hormone, ovulation, human chorionic gonadotropin.

scholarly journals Assessment of hemolytic effect of Cassia flower extracts on human RBCs

2018 ◽  
Vol 8 (6-s) ◽  
pp. 18-20 ◽  
Author(s):  
Shital S. Phuse ◽  
Zia. H. Khan
Keyword(s):  
Ethyl Acetate ◽  
Acetone Extract ◽  
In Vivo Studies ◽  
Rbc Membrane ◽  
Soxhlet Apparatus ◽  
Hemolytic Effect ◽  
Phosphate Buffered Saline ◽  
Human Rbcs

RBC membrane can be affected by consumption of bioactive compounds from herbs and medicinal plants. This study aimed to assess hemolytic effect of crude ethyl acetate and acetone extract from Cassia glauca flowers. Both the extracts of Cassia flowers were prepared, using Soxhlet apparatus. RBCs were washed with phosphate buffered saline and resuspended in 0.9% normal saline. These RBCs were added to different concentrations of the extracts and then incubated. After centrifugation, absorbance of the supernatant was determined by UV spectrophotometer at 540 nm. The present work shows that the fractions exhibited anti-hemolytic potential as extracts of Cassia flower showed very less percent of hemolysis when compared to standard quercetin. IC50values were found to be 23.77μg/ml for (CF EA) and 12.50μg/ml for Cassia flower in acetone(CF A)against standard which was found to be 41.75μg/ml. Extracts of Cassia flower exhibited very low hemolytic activity. Hence, it can be considered as safe to human RBCs.  In future recommend further in vitro and in vivo studies to evaluate the clinical efficacy of Cassia glauca extracts for treated several diseases. Keyword: Extract, Acetone, Ethyl acetate, Cassia glauca flowers, Hemolytic effect, RBCs

scholarly journals Demonstration of transferrin receptors on human placental trophoblast

Blood ◽  
1980 ◽  
Vol 55 (2) ◽  
pp. 240-242 ◽  
Author(s):  
GM Galbraith ◽  
RM Galbraith ◽  
A Temple ◽  
WP Faulk
Keyword(s):  
Binding Sites ◽  
Iron Transport ◽  
Transferrin Receptors ◽  
Transplacental Passage ◽  
Chorionic Villi ◽  
Characteristic Distribution ◽  
Maternal Iron ◽  
Phosphate Buffered Saline

Abstract It has been postulated that the transplacental passage of maternal iron to the developing fetus requires binding of maternal transferrin to the trophoblast. We have therefore examined the ability of the human placenta to bind transferrin in vitro. Transferrin was demonstrated on trophoblast of human chorionic villi by immunohistologic methods. Moreover, after removal of transferrin bound in vivo by treatment of tissue with chaotropic solution or phosphate-buffered saline, freshly added transferrin was shown to bind in vitro in the same characteristic distribution. These findings suggest that placental iron transport is initiated by uptake of maternal transferrin iron to specific trophoblast binding sites.

scholarly journals Evaluation of Marine Diindolinonepyrane in Vitro and in Vivo: Permeability Characterization in Caco-2 Cells Monolayer and Pharmacokinetic Properties in Beagle Dogs

Marine Drugs ◽  
2019 ◽  
Vol 17 (12) ◽  
pp. 651 ◽  
Author(s):  
Ma ◽  
Guo ◽  
Elango ◽  
Bao ◽  
Wu
Keyword(s):  
Liquid Chromatography ◽  
Mass Spectrum ◽  
Tissue Distribution ◽  
High Performance ◽  
Intravenous Route ◽  
Pharmacokinetic Data ◽  
Beagle Dogs ◽  
Total Recovery

A marine fibrinolytic compound was studied for use in thrombolytic therapy. Firstly, the absorption and transportation characteristics of 2,5-BHPA (2,5-BHPA:2,5-Bis-[8-(4,8-dimethyl-nona-3,7-dienyl)-5,7-dihydroxy-8-methyl-3-keto-1,2,7,8-tertahydro-6H-pyran[a]isoindol-2-yl]-pentanoic acid, a novel pyran-isoindolone derivative with bioactivity isolated from a rare marine microorganism in our laboratory) in the human Caco-2 cells monolayer model were investigated. We collected 2,5-BHPA in the cells to calculate the total recovery, and its concentration was analyzed by LC/MS/MS (Liquid Chromatography/ Mass Spectrum/ Mass Spectrum). The results showed that 2,5-BHPA has low permeability and low total recoveries in the Caco-2 cells membrane. Pharmacokinetics and tissue distribution of 2,5-BHPA were investigated in beagle dogs using HPLC (High Performance Liquid Chromatography) after intravenous administration of three different doses (7.5, 5.0, 2.5 mg·kg−1). Pharmacokinetic data indicated that 2,5-BHPA fitted well to a two-compartment model. Elimination half-lives (T1/2) were 49 ± 2, 48 ± 2, and 49 ± 2 min, respectively; the peak concentrations (Cmax) were 56.48 ± 6.23, 48.63 ± 5.53, and 13.64 ± 2.76 μg·mL−1, respectively; clearance rates (CL) were 0.0062 ± 0.0004, 0.0071 ± 0.0008, and 0.0092 ±0.0006 L·min−1·kg−1, respectively; mean retention times (MRT) were 28.17 ± 1.16, 26.23 ± 0.35, and 28.66 ± 0.84 min, respectively. The low penetrability of 2,5-BHPA indicated that the intravenous route of administration is more appropriate than the oral route. Meanwhile, 2,5-BHPA showed a good pharmacokinetic profile in beagle dogs. The tissue distribution showed that 2,5-BHPA could quickly distribute into the heart, intestines, liver, stomach, spleen, lungs, testicles, urine, intestine, kidneys, brain, and feces. The concentration of 2,5-BHPA was higher in the liver and bile. Interestingly, 2,5-BHPA was detected in the brain. Taken together, the above results suggested that our work might be beneficial in the development of agents for thrombolytic treatment.

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