Effect of ovarian stimulation by different gonadotrophin treatments on in vivo and in vitro reproductive efficiency of rabbit does under high ambient temperature

Background: Specialized freeze-drying process is being used in the field for different thermostable vaccine preparation worldwide. The thermostability remains only in undiluted conditions. If dilution is made at the morning and used for the whole day, the vaccine efficacy is compromised at high ambient temperature. In this study, trehalose based specialized vaccine diluent was used to improve the stability of Peste des Petits Ruminants (PPR) vaccine in diluted condition. Methods: The available PPR vaccine was reconstituted with conventional diluent and with trehalose based test diluent. The diluted vaccine was kept at ambient temperature without maintaining any cool chain. Stability of diluted vaccine virus was further assessed in vivo and in vitro at different temperatures. Goats were vaccinated and Vero cells were infected with reconstituted vaccines and were assessed at 0, 3, 6, 9 and 24 hours post dilution. Antibody titer was measured and virus infectivity titer was determined in both cultured cell lysate and supernatant. The presence of the virus particles in Vero cell was confirmed by standard RT-PCR targeting Fusion (F) gene of PPR virus. Results: In vivo results revealed that the number of goats possessed antibodies to PPR virus was higher in trehalose based vaccine formulation than the conventional PBS based diluent. Reconstituted vaccine virus (using PBS and trehalose diluent) infected Vero cells produced 70-80% cytopathic effect (CPE) in 5th days of post infection. Both diluents produced and maintained infectivity titer from log10 TCID50 5.5 to log10 TCID50 3.6, until the use of vaccines incubated for 9 hours after dilution. On the other hand, at 24 hours of post dilution only trehalose formulated vaccine produced log10 TCID502.5 whereas no infectivity titer was observed at the same time using conventional one. Conclusion: The present study suggests that trehalose preserves the quality of reconstituted vaccine in terms of infectivity titers. Trehalose can be a diluent of choice for reconstitution of PPR vaccine in field.

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314 MELATONIN ON IN VIVO AND IN VITRO MATURATION OF MOUSE OOCYTES

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab314 ◽

2015 ◽

Vol 27 (1) ◽

pp. 246 ◽

Cited By ~ 1

Author(s):

H. Fernandes ◽

L. Schefer ◽

F. C. Castro ◽

C. L. V. Leal

Keyword(s):

Ovarian Stimulation ◽

In Vitro Maturation ◽

F1 Hybrids ◽

Saline Control ◽

Control Group ◽

Mouse Oocytes ◽

Maturation Rate ◽

Different Doses

Melatonin is a pineal hormone related to the control of the circadian cycle, besides the reproductive seasonality of some animal species, and has shown positive effects on oocyte maturation and embryo development. The aim of this study was to assess the effects of melatonin on in vivo and in vitro maturation of mouse oocytes. Female F1 hybrids (C57BL/6 × CBA; n = 8 per group/treatment) were used in 3 different treatments (trt) groups: (I) in vivo trt: mice received 2 different doses of melatonin injections, 10 and 20 mg kg–1 per IP including a saline control dose (0 mg kg–1 per IP) for 4 days along with ovarian stimulation trt of 5 IU of eCG IP, followed by 5 IU of hCG IP 48 h later, and cumulus-oocyte complexes (COC) were collected 16 h after hCG; (II) mice received a similar in vivo melatonin trt, but ovarian stimulation trt was only 5 IU of eCG, no hCG, and COC were collected after 48 h and subsequently matured in vitro with 0.5 µg mL–1 of FSH for 16 h; (III) in vitro maturation of oocytes: COC were collected 48 h after 5 IU of eCG and maturated in the presence of 3 different doses of melatonin (10–9, 10–6, and 10–3 M) or 0.5 µg mL–1 of FSH (control) for 16 h. In vitro-maturing oocytes were in incubated at 37°C, 5% CO2, and 95% humidity. Maturation rates were evaluated according to the presence of the first polar body under an inverted microscope. Statistical analyses were performed by ANOVA followed by Tukey's test (4 replicates). In the first treatment, 20 mg kg–1 of melatonin showed the highest in vivo maturation rate, 80.3% (61/76), while 10 mg kg–1 of melatonin was 62.4% (53/85) and the saline control group was 69.4% (77/111), but differences were not significant (P > 0.05). For in vitro maturation of oocytes from animals previously treated with melatonin, the 10 mg kg–1 of melatonin group had the highest maturation rate, 53.2% (99/186), in comparison with the saline and 20 mg kg–1 of melatonin groups, which showed 46.6 (88/189) and 39.0% (85/218), respectively; again, no differences were detected (P > 0.05). In the last treatment, the maturation rates increased from 48.9 (43/88) to 53.7 (51/95) and 56.0% (56/100) as the melatonin concentrations decreased from 10–3, 10–6, and 10–9 M, respectively. The control group had the highest rate of 57.3% (55/96), but no statistical differences were observed (P = 0.706). In conclusion, under the conditions studied, melatonin was unable to improve the maturation rate neither after in vivo nor in vitro treatment. However, during in vitro maturation, melatonin alone was as efficient as FSH in promoting maturation in murine oocytes, indicating its potential effect on stimulating meiosis. Therefore, the role of melatonin in stimulating meiosis needs further investigation.Acknowledgments to FAPESP for fellowship (HF) and funding (CLVL).

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The Parkes lecture: controlled ovarian stimulation in women

Reproduction ◽

10.1530/reprod/120.2.201 ◽

2000 ◽

pp. 201-210 ◽

Cited By ~ 21

Author(s):

SG Hillier

Keyword(s):

Ovarian Stimulation ◽

Ovarian Function ◽

Recombinant Dna ◽

New Drugs ◽

Controlled Ovarian Stimulation ◽

Recombinant Dna Technology ◽

Vitro Fertilization ◽

Ovarian Folliculogenesis

Recent advances in knowledge of the endocrine and paracrine mechanisms that regulate human ovarian folliculogenesis have been parallelled by the introduction into clinical practice of new drugs that can be used safely and effectively to stimulate ovarian function in infertile women. Most notably, recombinant DNA technology has been applied to the production of molecularly pure forms of the gonadotrophins, FSH and LH, opening the way to the development of improved strategies for manipulating the ovarian paracrine system. The clinical objectives of controlled ovarian stimulation fall into two categories, depending on patient needs: (1) induction of multiple follicles from which mature oocytes can be harvested for use in assisted reproduction protocols such as in vitro fertilization and embryo transfer; or (2) induction of spontaneous ovulation of a single mature follicle so that conception might occur in vivo. This review summarizes the physiological principles upon which the use of gonadotrophins for clinical purposes is based, highlighting new opportunities for improved treatment as a result of the availability of recombinant FSH and LH.

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In vivo environmental temperature and the in vitro pattern of luminal acidification in turtle bladders. Evidence for HCO3 ion reabsorption.

The Journal of General Physiology ◽

10.1085/jgp.92.5.613 ◽

1988 ◽

Vol 92 (5) ◽

pp. 613-642 ◽

Cited By ~ 2

Author(s):

T P Schilb ◽

J H Durham ◽

W A Brodsky

Keyword(s):

Physical Activity ◽

Food Intake ◽

Ambient Temperature ◽

Environmental Temperature ◽

Acid Excretion ◽

Turtle Bladder ◽

Single Temperature

In this study, it is shown how to transfer tared aliquots of (HCO3 + CO2)-containing luminal fluids directly into the mercury-sealed chamber of a modified Van Slyke apparatus and how to obtain direct as well as indirect manometric determinations of dissolved CO2 ([CO2]f) in each aliquot of such fluids. It is next shown that the pattern of in vitro luminal acidification in an isolated turtle bladder sac depends upon the prior in vivo ambient temperature to which the donor turtle had become adapted. Under in vivo conditions, the food intake, physical activity, and acid excretion of 32 degrees C-adapted turtles are greater than those of 21 degrees C or 26 degrees C-adapted turtles. Under in vitro conditions of incubating isolated bladder sacs (from 21, 26, and 32 degrees C turtles) in (HCO3 + CO2)-containing Ringer media at a single temperature (21 degrees C), the patterns of luminal acidification are as follows: (a) The rate of depletion of luminal [HCO3] is greatest in bladders from the 32 degrees C-adapted turtles. (b) Concomitant decreases in luminal [CO2]f, [HCO3], and pH (the 'CO2-decreasing patterns' of luminal acidification) develop in all bladders from 32 degrees C turtles, in half of those from 26 degrees C turtles, but in less than one-fifth of those from 21 degrees C-adapted turtles: and (c) a CO2-increasing pattern of luminal acidification is found in most of the bladders from 21 degrees C-adapted turtles. A postulated bicarbonate ion-reabsorbing pump is consistent with all of these patterns of luminal acidification.

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SPAs promote thermomorphogenesis via regulating the phyB-PIF4 module in Arabidopsis

10.1101/2020.02.07.938951 ◽

2020 ◽

Author(s):

Sanghwa Lee ◽

Inyup Paik ◽

Enamul Huq

Keyword(s):

Ambient Temperature ◽

Kinase Activity ◽

Global Gene Expression ◽

High Ambient Temperature ◽

Fine Tuning ◽

Plant Growth And Development ◽

Profound Influence ◽

Hyponastic Growth

SUMMARYHigh ambient temperature due to global warming has a profound influence on plant growth and development at all stages of life cycle. Plant response to high ambient temperature termed thermomorphogenesis is characterized by hypocotyl and petiole elongation, and hyponastic growth at seedling stage. However, the molecular mechanism of thermomorphogenesis is still rudimentary. Here, we show that a set of four SUPPRESSOR OF PHYA-105 (SPA) genes are required for thermomorphogenesis. Consistently, SPAs are necessary for global gene expression changes in response to high ambient temperature. SPA1 level is unaffected, while the thermosensor phyB is stabilized in the spaQ mutant at high ambient temperature. Furthermore, in the absence of four SPA genes, the pivotal transcription factor PIF4 fails to accumulate, indicating a role of SPAs in regulating the phyB-PIF4 module at high ambient temperature. SPA1 directly phosphorylates PIF4 in vitro, and a mutant SPA1 affecting the kinase activity fails to rescue the PIF4 level as well as the thermo-insensitive phenotype of spaQ, suggesting that the SPA1 kinase activity is necessary for thermomorphogenesis. Taken together, these data suggest that SPAs integrate light and temperature signaling via fine tuning the phyB-PIF4 module.

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Generation of UCiPSCs-Derived Neurospheres for Cell Therapy and its Application in Cell Shipment at Ambient Temperature

10.21203/rs.3.rs-47992/v1 ◽

2020 ◽

Author(s):

Shuai Li ◽

Huifang Zhao ◽

Xiaobo Han ◽

Lang He ◽

Omar Mukama ◽

...

Keyword(s):

Stem Cells ◽

Cell Therapy ◽

Neural Stem Cells ◽

Ambient Temperature ◽

Low Cost ◽

Survival Rates ◽

High Survival ◽

Simple Method

Abstract BackgroundNeural stem cells（NSCs）therapy remains one of the most potential approaches for neurological disorders treatment. The discovery of human induced pluripotent stem cells (hiPSCs) and the establishment of hiPSC-derived human neural stem cells (hiNSCs) have revolutionized our technique to cell therapy. Meanwhile, it is often required that NSCs are stored and transported long distances for research or treatment. Although high survival rates could be maintained, conventional methods of cell transport (dry ice or liquid nitrogen) are inconvenient and expensive. Therefore，the establishment of a safe, affordable, and frequent obtained hiPSCs and hiNSCs, with characteristics that match fetal hNSCs and a simple, low-cost way to store and transport, are incredibly urgent. MethodsWe reprogrammed human urinary cells to iPSCs using a virus-free technique and differentiated the iPSCs toward iNSCs/neurospheres and neurons, under Good Manufacturing Practice (GMP)-compatible conditions. The pluripotency of iPSCs and iNSCs was characterized by a series of classical methods (surface markers, karyotype analysis and in vitro and in vivo differentiation capabilities, etc).ResultsHere, our results showed that we successfully generated hiNSCs/neurospheres from more available, non-invasive, and more acceptable urinary cells by a virus-free technique and their differentiation into neural networks. Moreover，hiNSCs survived longer as neurospheres at ambient temperature than those cultured in a monolayer. Approximately 7 days, the neural viability remained at > 80%, while hiNSCs cultured in a monolayer died almost immediately. Neurospheres exposed to ambient temperature that were placed under standard culture conditions (37 ℃, 5% CO2) recovered their typical morphology, and retained their ability to proliferate and differentiate. ConclusionsIn this study, we provided a simple method for the storage of NSCs as neurospheres at ambient temperature as an alternative to more costly and inconvenient traditional methods of cryopreservation. This will enable hiNSCs to be transported over long distances at ambient temperature and facilitate the therapeutic application of NSCs as neurospheres without any further treatment.

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Fertility of boars - what is important to know

Biotechnology in Animal Husbandry ◽

10.2298/bah1702135s ◽

2017 ◽

Vol 33 (2) ◽

pp. 135-149 ◽

Cited By ~ 3

Author(s):

Radomir Savic ◽

Raquel Marcos ◽

Milica Petrovic ◽

Dragan Radojkovic ◽

Cedomir Radovic ◽

...

Keyword(s):

Litter Size ◽

Return Rate ◽

Low Fertility ◽

Reproductive Efficiency ◽

Reproductive Management ◽

Fertility Traits ◽

Farrowing Rate ◽

Sperm Traits

The most important part in reproductive management is the control of boar fertility. A common division of fertility traits is on the: in vitro (sperm traits) and in vivo (return rate, farrowing rate and litter size traits) fertility. In many studies were found differences between breed in the both groups of fertility traits. Variability of sperm traits of boars during the reproductive exploitation is influenced by various genetic (boar, breed) and paragenetic factors/effects (age, season, intensity of use). Good libido is desirable characteristics in boars, but the knowlegde of the correlation of libido and boar fertility traits are limited. Also, there is no standardised procedure or methods for the estimation of libido of the boars. The permanent ranking of boars according to the reproductive efficiency should be performing. Good reproductive management implies the timely identification of boars with the low fertility (or close to the average).

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