scholarly journals Immunization with CENP-C Causes Aberrant Chromosome Segregation during Oocyte Meiosis in Mice

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Jiao Fan ◽  
Yang Liu ◽  
Yiping Zhong
Keyword(s):  
Protein C ◽  
Polar Body ◽  
Control Group ◽  
Human Centromere ◽  
Centromere Protein ◽  
Oocyte Meiosis ◽  
First Polar Body ◽  
Freund’S Adjuvant ◽  
Freund's Adjuvant ◽  
Experimental Group

Anticentromere antibodies (ACA) were associated with lower oocyte maturation rates and cleavage rates, while the mechanism was not clear. Aims of this study were to examine whether active immunization with centromere protein C could elicit the CENP-C autoantibody in mice and the impacts of the CENP-C autoantibody on oocyte meiosis. Mice were divided into two groups, one was the experimental group immunized with human centromere protein C and Freund’s adjuvant (CFA), and the other was the control group injected with CFA only. Serum and oocytes of BALB/c mice immunized with human centromere protein C (CENP-C) in complete Freund’s adjuvant (CFA) or injected with only CFA were studied for the development of the CENP-C antibody. Rates of germinal vesicle breakdown (GVBD), first polar body (Pb1) extrusion, abnormal spindle morphology, and chromosome misalignment were compared between the experimental group and the control group. The CENP-C antibody was only observed in serum and oocytes of mice immunized with the centromere protein C antigen. The first polar body (Pb1) extrusion rate was lower in the experimental group ( P < 0.01 ). A higher percentage of spindle defects and chromosome congression failure were also detected in the experimental group (spindle defects: 64.67 ± 1.16 % vs. 9.27 ± 2.28 % control; chromosome misalignment: 50.80 ± 2.40 % vs. 8.30 ± 1.16 % control; P < 0.01 for both). Oocyte meiosis was severely impaired by the CENP-C antibody, which may be the main mechanism of adverse reproductive outcomes for ACA-positive women who have no clinical symptoms of any autoimmune diseases.

scholarly journals Bushen Tiaojing (II and III) Decoctions Activate MAPK14 and MAPK3/1 to Promote the Expression of Cumulus Expansion-Related Factors in Mice

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Xiao Liang ◽  
Xue Tong ◽  
Hui-lan Du ◽  
Ming He ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Western Blot ◽  
Polar Body ◽  
Cumulus Cell ◽  
Serum Levels ◽  
Control Group ◽  
Distilled Water ◽  
Related Factors ◽  
Cumulus Expansion ◽  
Sequential Administration ◽  
First Polar Body

Background. Bushen Tiaojing Decoctions (BSTJ-II-D and BSTJ-III-D) are used to assist pregnancy in clinical practice. In this study, we explored the ability of sequential administration of BSTJ-II-D and BSTJ-III-D to promote cumulus cell (CC) expansion and its underlying mechanisms in controlled ovarian hyperstimulation (COH) mice. Methods. Kunming mice were randomly divided into three groups. The normal group was injected intraperitoneally with saline, and distilled water was administered orally by gavage. As the COH model, mice were injected with GnRHa, eCG, and hCG. Subsequently, the BSTJD group received BSTJ-II-D and BSTJ-III-D orally by gavage, while the control group received distilled water. We evaluated CC expansion and oocyte first polar body (PB1) extrusion under a stereomicroscope. Serum levels of follicle-stimulating hormone (FSH) were detected by radioimmunoassay. The expression of the CC expansion-related factors PTX3 and PTGS2 was detected by immunofluorescence, western blot, and quantitative real-time-polymerase chain reaction analyses (qRT-PCR). Expression of p-MAPK14, p-MAPK3/1, MAPK14, and MAPK3/1 was detected by western blot analysis. Results. Sequential administration of BSTJ-II-D and BSTJ-III-D promoted cumulus expansion and oocyte PB1 extrusion and upregulated PTX3 and PTGS2 expression at the mRNA and protein levels. Furthermore, the levels of p-MAPK14/MAPK14, p-MAPK3/1/MAPK3/1 proteins, and serum FSH in the BSTJD group were higher than those in the normal and control groups. Conclusions. Sequential administration of BSTJ-II-D and BSTJ-III-D promotes cumulus expansion and oocyte maturation in COH mice by increasing FSH expression and activating the MAPK14 and MAPK3/1 signalling pathways, thereby increasing expression of PTX3 and PTGS2.

H1foo is essential for in vitro meiotic maturation of bovine oocytes

Zygote ◽  
2014 ◽  
Vol 23 (3) ◽  
pp. 416-425 ◽  
Author(s):  
Yan Yun ◽  
Peng An ◽  
Jing Ning ◽  
Gui-Ming Zhao ◽  
Wen-Lin Yang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Linker Histone ◽  
Meiotic Maturation ◽  
Control Group ◽  
Bovine Oocyte ◽  
First Polar Body ◽  
Effective Sirnas ◽  
Bovine Oocytes

SummaryOocyte-specific linker histone, H1foo, is localized on the oocyte chromosomes during the process of meiotic maturation, and is essential for mouse oocyte maturation. Bovine H1foo has been identified, and its expression profile throughout oocyte maturation and early embryo development has been established. However, it has not been confirmed if H1foo is indispensable during bovine oocyte maturation. Effective siRNAs against H1foo were screened in HeLa cells, and then siRNA was microinjected into bovine oocytes to down-regulate H1foo expression. H1foo overexpression was achieved via mRNA injection. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that H1foo was up-regulated by 200% and down-regulated by 70%. Based on the first polar body extrusion (PB1E) rate, H1foo overexpression apparently promoted meiotic progression. The knockdown of H1foo significantly impaired bovine oocyte maturation compared with H1foo overexpression and control groups (H1foo overexpression = 88.7%, H1foo siRNA = 41.2%, control = 71.2%; P < 0.05). This decrease can be rescued by co-injection of a modified H1foo mRNA that has escaped from the siRNA target. However, the H1e (somatic linker histone) overexpression had no effect on PB1E rate when compared with the control group. Therefore we concluded that H1foo is essential for bovine oocyte maturation and its overexpression stimulates the process.

scholarly journals Berberine Delays Onset of Collagen-Induced Arthritis through T Cell Suppression

2021 ◽  
Vol 22 (7) ◽  
pp. 3522
Author(s):  
Alexandra A. Vita ◽  
Hend Aljobaily ◽  
David O. Lyons ◽  
Nicholas A. Pullen
Keyword(s):  
T Cell ◽  
Type Ii Collagen ◽  
Control Group ◽  
Collagen Induced Arthritis ◽  
Preclinical Phase ◽  
T Cell Suppression ◽  
Cell Suppression ◽  
Freund’S Adjuvant ◽  
Freund's Adjuvant

There is evidence that berberine (BBR), a clinically relevant plant compound, ameliorates clinically apparent collagen-induced arthritis (CIA) in vivo. However, to date, there are no studies involving the use of BBR which explore its prophylactic potential in this model of rheumatoid arthritis (RA). The aim of this study was to determine if prophylactic BBR use during the preclinical phase of collagen-induced arthritis would delay arthritic symptom onset, and to characterize the cellular mechanism underlying such an effect. DBA/1J mice were injected with an emulsion of bovine type II collagen (CII) and complete Freund’s adjuvant (day 0) and a booster injection of CII in incomplete Freund’s adjuvant (day 18) to induce arthritis. Mice were then given i.p. injections of 1 mg/kg/day of BBR or PBS (vehicle with 0.01% DMSO) from days 0 to 28, were left untreated (CIA control), or were in a non-arthritic control group (n = 15 per group). Incidence of arthritis in BBR-treated mice was 50%, compared to 90% in both the CIA and PBS controls. Populations of B and T cells from the spleens and draining lymph nodes of mice were examined on day 14 (n = 5 per group) and day 28 (n = 10 per group). BBR-treated mice had significantly reduced populations of CD4+Th and CD4+CXCR5+ Tfh cells, and an increased proportion of Foxp3+ Treg at days 14 and 28, as well as reduced expression of co-stimulatory molecules CD28 and CD154 at both endpoints. The effect seen on T cell populations and co-stimulatory molecule expression in BBR-treated mice was not mirrored in CD19+ B cells. Additionally, BBR-treated mice experienced reduced anti-CII IgG2a and anti-CII total IgG serum concentrations. These results indicate a potential role for BBR as a prophylactic supplement for RA, and that its effect may be mediated specifically through T cell suppression. However, the cellular effector involved raises concern for BBR prophylactic use in the context of vaccine efficacy and other primary adaptive immune responses.

scholarly journals FUNCTIONAL ANALYSIS OF AGERATUM CONYZOIDES L. (BABANDOTAN) LEAVES EXTRACT ON RHEUMATOID ARTHRITIS MODEL RAT

2017 ◽  
Vol 10 (3) ◽  
pp. 429
Author(s):  
Erna Harfiani ◽  
Riri Nurul Suci ◽  
Ade Arsianti ◽  
Anton Bahtiar ◽  
Katrin Basah
Keyword(s):  
Rheumatoid Arthritis ◽  
Normal Control ◽  
Rat Model ◽  
Control Group ◽  
Paw Edema ◽  
Ageratum Conyzoides ◽  
Tnf Α ◽  
Freund’S Adjuvant ◽  
Complete Freund's Adjuvant ◽  
Freund's Adjuvant

ABSTRACTObjective: Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation in joints. Ageratum conyzoides L. (Babandotan)leaves are proven to be used in inflammation therapy, yet there is a little data regarding the effects of the leaves on RA. The aim of this study is toinvestigate anti-RA activity of the ethanolic extract of A. conyzoides L. leaves (EEAL) harvested from Bogor, Indonesia, in rats.Methods: The phytochemical screening analysis and thin-layer chromatography were performed to analyze the constituents of the EEAL. This studyused white male Sprague Dawley rats which were divided into 6 groups; normal control and negative control groups, both given 0.5% carboxymethylcellulose; the positive control group, given methotrexate suspension (0.05 mg/200 g bw.); the dose variation extract is 40 mg, 80 mg, and 160 mg/200 gbody weight. All the groups were induced with 0.1 ml Complete Freund’s adjuvant on day 1, except normal control group. Some parameters will bemeasured, such as paw edema, levels of leukocyte and lymphocyte, concentrations of tumor necrosis factor-α (TNF-α), and the number of osteoclastsper mm2.Results: For 21 days the rats have given treated the EEAL in three doses, was showed the decreasing volume of paw edema, levels of blood leukocytesand lymphocytes, concentrations of TNF-α and the number of osteoclasts, compared to the rat model of RA.Conclusion: This study showed that the leaves of A. conyzoides L. harvested from Bogor, Indonesia, have activity as anti-RA in a rat model, in whichflavonoid plays a role in inhibition of chronic inflammatory processes.Keywords: Ageratum conyzoides L. leaves, Anti-rheumatoid arthritis, Complete Freund’s adjuvant, Flavonoids.

40 TRICHOSTATIN A TREATMENT EFFECTS ON IN VITRO DEVELOPMENT OF INTERSPECIES NUCLEAR TRANSFER CAT EMBRYOS DEPEND ON RECIPIENT CYTOPLASM SPECIES

2015 ◽  
Vol 27 (1) ◽  
pp. 113
Author(s):  
L. T. K. Do ◽  
Y. Sato ◽  
M. Taniguchi ◽  
T. Otoi
Keyword(s):  
Nuclear Transfer ◽  
Treatment Effects ◽  
Trichostatin A ◽  
Polar Body ◽  
Blastocyst Stage ◽  
Control Group ◽  
Fluid Medium ◽  
First Polar Body ◽  
Bovine Oocytes

The developmental ability of interspecies somatic cell nuclear transfer (iSCNT) embryos decreases as the taxonomic distance between the donor and recipient species increases. Treatment of cat iSCNT embryos using bovine oocytes with 50 nM of trichostatin A (TSA) improves in vitro embryonic development (Wittayarat et al. 2013 Cell. Reprogram. 15, 301–308). This study investigated whether the TSA treatment effects differ between the development of cat iSCNT embryos reconstructed with porcine and bovine oocytes. Porcine and bovine cumulus-oocyte complexes were in vitro matured for 44 h and 24 h, respectively. After cumulus cell removal, enucleation was performed by aspiration of the metaphase II plate and the first polar body using a piezo-driven pipette. A cat fibroblast cell was then injected into cytoplasm of successfully enucleated oocyte. Reconstructed cybrids were electrically activated by a single 1.5 kV cm–1 pulse for 100 µs (pig-cat embryos), or a 2.3 kV cm–1 pulse for 30 µs (cow-cat embryos). Pig-cat and cow-cat embryos were cultured in porcine zygote medium (PZM)-5 and modified synthetic oviducal fluid medium (mSOF), respectively. After electrical activation, pig-cat and cow-cat embryos were cultured in medium supplemented with 5 µg mL–1 cytochalasin B + 50 nM TSA (TSA group) or without TSA (control group), and the cow-cat embryo medium was also supplemented with 10 µg mL–1 cycloheximide. After 2 h, TSA-treated pig-cat and cow-cat embryos were incubated in medium supplemented with TSA for 22 h, followed by 48 h incubation without TSA. Pig-cat and cow-cat control embryos were cultured in medium without TSA for 70 h after activation. Then, all pig-cat and cow-cat embryos were cultured in porcine blastocyst medium (PBM) or mSOF medium supplemented with 5% fetal bovine serum, respectively, for 5 additional days. Four to seven replicates were performed for each experiment. Data were analysed using Student's t-test. For pig-cat embryos, no difference was observed in cleavage rates between both groups, but development to the blastocyst stage was higher in the pig control group (n = 147, 8.0%) than that of pig TSA group (n = 131, 0.7%; P < 0.05). In contrast, development to the blastocyst stage in cow-cat embryos was not observed in the cow control group (n = 125, 0%), but it was observed in cow TSA group (n = 136, 3.7%). These results indicate that TSA treatment effects are species-specific, but those effects remain to be clarified.

Mutational Analysis of the Central Centromere Targeting Domain of Human Centromere Protein C, (CENP-C)

2002 ◽  
Vol 275 (1) ◽  
pp. 81-91 ◽  
Author(s):  
Kang Song ◽  
Bobbi Gronemeyer ◽  
Wei Lu ◽  
Emily Eugster ◽  
John E. Tomkiel
Keyword(s):  
Protein C ◽  
Mutational Analysis ◽  
Human Centromere ◽  
Centromere Protein

scholarly journals Vitamin C protects porcine oocytes from microcystin-LR toxicity during maturation

2020 ◽  
Author(s):  
Xue Zhang ◽  
Changyin Zhou ◽  
Weijian Li ◽  
Juan Li ◽  
Wangjun Wu ◽  
...  
Keyword(s):  
Dna Damage ◽  
Vitamin C ◽  
Actin Polymerization ◽  
Polar Body ◽  
Cumulus Cell ◽  
Human Reproduction ◽  
Secondary Products ◽  
Oocyte Meiosis ◽  
First Polar Body ◽  
The Impact

Abstract Background: Microcystin-leucine arginine (MC-LR) is the most toxic cyanotoxin found in water bodies. Microcystins are produced as secondary products of cyanobacteria metabolism. They have a stable structure, and can bioaccumulate in living organisms. Humans and livestock who drink fresh water containing MC-LR can be poisoned. However, few studies have reported the effects of MC-LR exposure on livestock or human reproduction.Results: We used porcine oocytes as the model to explore the effects of MC-LR on oocyte maturation, and studied the impact of vitamin C (VC) administration on MC-LR‐induced meiosis defects. Exposure to MC-LR significantly restricted cumulus cell expansion and decreased first polar body extrusion. Further studies showed that MC-LR exposure led to meiosis arrest by disturbing cytoskeleton dynamics with MC-LR exposed oocytes displaying aberrant spindle organization, low levels of acetylate α‐tubulin, and disturbed actin polymerization. Additionally, MC-LR exposure impaired cytoplasmic maturation by disturbing mitochondria distribution. Moreover, MC-LR also produced abnormal epigenetic modifications, and induced high levels of oxidative stress and DNA damage. The administration of VC provided partial protection from all of the defects observed in oocytes exposed to MC-LR. Conclusions: These results demonstrate that MC-LR has a toxic effect on oocyte meiosis through the generation of excessive ROS levels and DNA damage. Supplementation of VC was able to protect against MC-LR-induced oocyte damage and represents a potential therapeutic strategy to improve the quality of MC-LR-exposed oocytes.

scholarly journals ERDOSTEINE: AN EFFECTIVE ANTIOXIDANT FOR PROTECTING COMPLETE FREUND’S ADJUVANT INDUCED ARTHRITIS IN RATS

2021 ◽  
pp. 71-75
Author(s):  
BANYLLA SYNMON ◽  
SANHATIDUTTA ROY ◽  
SUTAPA BISWAS MAJEE ◽  
MEGHNA PAUL ◽  
SANDIPAN DASGUPTA
Keyword(s):  
Body Weight ◽  
Test Group ◽  
The Body ◽  
Complete Freund’S Adjuvant ◽  
Control Group ◽  
Albino Rats ◽  
Standard Group ◽  
Freund’S Adjuvant ◽  
Complete Freund's Adjuvant ◽  
Freund's Adjuvant

Objective: The objective of this study was to evaluate the protective effect of Erdosteine on complete freund’s adjuvant (CFA) induced arthritic rats. Methods: Wistar Albino rats of 100–250 g were divided into five groups (n=6) and administered with 0.1 ml of CFA subcutaneously into the left hind paw except the negative control group. The standard group received methotrexate (MTX) 0.075 mg/kg body weight orally. Besides, the test groups received Erdosteine orally at a dose 10 mg/kg and 20 mg/kg bodyweight for 12 days. The changes in body weight, paw volume, hematological parameters, radiographical, and histological findings were the indicators to evaluate the efficacy of the test product. Discussion: Significant change in the body weight, paw volume, radiographical, hematological, and histological parameters were observed which supports the remarkable reduction of the arthritic development in the standard and test groups compared to the untreated group. However, the test group (Erdosteine) with the dose 20 mg/kg shows to be more potent than the test group (Erdosteine) with a dose 10 mg/kg and the standard group (MTX) to reduce the arthritic effect. Results: The test group with 20 mg/kg Erdosteine showed much better outcome than the standard group at significant (p<0.05). Therefore, Erdosteine acting as an anti-inflammatory and anti-oxidant is effective at a dose 20 mg/kg in treating the progression of rheumatoid arthritis in rats.

scholarly journals Pengaruh urea dalam media maturasi in vitro terhadap tingkat maturasi oosit sapi

2021 ◽  
Vol 10 (2) ◽  
pp. 46
Author(s):  
Sepvian Dewi Kurniawati ◽  
Suryanie Sarudji ◽  
Widjiati Widjiati
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Metaphase Plate ◽  
Petri Dish ◽  
Control Group ◽  
Maturation Medium ◽  
First Polar Body ◽  
Maturation Rate ◽  
Follicle Aspiration

This study was aimed to determine the effect of urea in maturation medium on in vitro oocyte maturation rate. The medium used was TCM-199 added with Hepes, NaHCO3, Kanamycin 0.15 IU/mL, PMSG, 0.15 IU/mL hCG, and 10% FBS. Cumulus oocyte complexes (COCs) of cows derived from follicle aspiration were divided into three groups. In control group (P0), the COCs were matured in vitro in a maturation medium without urea addition, meanwhile in the P1 and P2 groups, the medium was added with urea 20 and 40 mg/dL, respectively. Each petri dish contained three drops of maturation medium (300 µl/drops) according to the groups. Microdrops were coated with mineral oil and then incubated in a 5% CO2 incubator, at 39 ˚C with maximum humidity. Aceto-orcein staining was conducted to evaluate the maturation of oocytes based on the achievement of metaphase II phase that is indicated by the presence of metaphase plate and/or first polar body. The result showed that the oocyte maturation rates of P0, P1, and P2 were 51.25, 52.43 (p >0.05), and 46.88 % (p <0.05) respectively. It could be concluded that the presence of urea at 40 mg/dL in maturation medium reduced the percentage of bovine oocyte maturation in vitro.

scholarly journals Evidence for anti-viral effects of complete Freund’s adjuvant in the mouse model of enterovirus infection

2020 ◽  
Author(s):  
Arunakumar Gangaplara ◽  
Chandirasegaran Massilamany ◽  
Ninaad Lasrado ◽  
David Steffen ◽  
Jay Reddy
Keyword(s):  
Viral Infections ◽  
Complete Freund’S Adjuvant ◽  
Control Group ◽  
Cell Responses ◽  
Freund’S Adjuvant ◽  
Complete Freund's Adjuvant ◽  
Freund's Adjuvant ◽  
Group B ◽  
Viral Protein 1 ◽  
Adjuvant Effects

AbstractGroup B Coxsackieviruses belonging to the genus, Enterovirus, contain six serotypes that induce various diseases, whose occurrence may involve the mediation of more than one serotype. We recently identified immunogenic epitopes within CVB3 viral protein 1 that induce anti-viral T cell responses in mouse models of CVB infections. In our investigations to determine the protective responses of the viral epitopes, we unexpectedly noted that animals immunized with complete Freund’s adjuvant (CFA) alone and later challenged with CVB3 were completely protected against myocarditis. Similarly, the pancreatitis-inducing ability of CVB3 was remarkably reduced to only 10% in the CFA group as opposed to 73.3% in the control group that received no CFA. Additionally, no mortalities were noted in the CFA group, whereas 40% of control animals died during the course of 21 days post-infection with CVB3. Taken together, our data suggest that the adjuvant effects of CFA may be sufficient for protection against CVB infections. These observations may provide new insights into our understanding of the occurrence of viral infections. One example is Coronavirus disease-19 (COVID-19) as individuals suffering from COVID-19 who have been vaccinated with Bacillus Calmette–Guérin appear to have fewer morbidities and mortalities than unvaccinated individuals.

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