Leptin Reduces Plin5 m6A Methylation through FTO to Regulate Lipolysis in Piglets

Perilipin5 (Plin5) is a scaffold protein that plays an important role in lipid droplets (LD) formation, but the regulatory effect of leptin on it is unclear. Our study aimed to explore the underlying mechanisms by which leptin reduces the N6-methyladenosine (m6A) methylation of Plin5 through fat mass and obesity associated genes (FTO) and regulates the lipolysis. To this end, 24 Landrace male piglets (7.73 ± 0.38 kg) were randomly sorted into two groups, either a control group (Control, n = 12) or a 1 mg/kg leptin recombinant protein treatment group (Leptin, n = 12). After 4 weeks of treatment, the results showed that leptin treatment group had lower body weight, body fat percentage and blood lipid levels, but the levels of Plin5 mRNA and protein increased significantly in adipose tissue (p < 0.05). Leptin promotes the up-regulation of FTO expression level in vitro, which in turn leads to the decrease of Plin5 M6A methylation (p < 0.05). In in vitro porcine adipocytes, overexpression of FTO aggravated the decrease of M6A methylation and increased the expression of Plin5 protein, while the interference fragment of FTO reversed the decrease of m6A methylation (p < 0.05). Finally, the overexpression in vitro of Plin5 significantly reduces the size of LD, promotes the metabolism of triglycerides and the operation of the mitochondrial respiratory chain, and increases thermogenesis. This study clarified that leptin can regulate Plin5 M6A methylation by promoting FTO to affect the lipid metabolism and energy consumption, providing a theoretical basis for treating diseases related to obesity.

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Mechanism of the adverse effect of hyaluronidase used for oocyte denudation on early development of bovine embryos

Zygote ◽

10.1017/s0967199421000010 ◽

2021 ◽

pp. 1-5

Author(s):

Shiori Ashibe ◽

Kanade Irisawa ◽

Ken Yokawa ◽

Yoshikazu Nagao

Keyword(s):

Treatment Group ◽

Assisted Reproductive Technologies ◽

Cumulus Cells ◽

Reproductive Technologies ◽

Control Group ◽

Free Calcium ◽

Parthenogenetic Development ◽

Early Embryos ◽

Bovine Oocytes

Summary Hyaluronidase is widely used in animal and human assisted reproductive technologies (ARTs) to remove cumulus cells around oocytes. However, adverse effects of hyaluronidase treatment, such as increased rates of degeneration and parthenogenesis, have been found after treatment of human and mouse oocytes. Currently, the mechanism(s) of the detrimental effects are unclear. The present study was initiated to identify the mechanism of adverse responses to hyaluronidase treatment in bovine oocytes and early embryos. Cumulus cells were removed from cumulus–oocyte complexes (COCs) with or without hyaluronidase and the oocytes were subjected to intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Significantly lower rates of blastocyst formation were obtained in the hyaluronidase treatment group after ICSI (22.4%) and IVF (21.2%) compared with the non-hyaluronidase control groups: 36.1% after ICSI and 30.4% after IVF. Next, we examined the effect of hyaluronidase on parthenogenetic development rates and on the cytoplasmic levels of free calcium ions (Ca2+), reactive oxygen species (ROS) and reduced glutathione (GSH). No differences in parthenogenesis rates were found between treated and untreated groups. Ca2+ levels in oocytes from the hyaluronidase treatment group indicated using mean fluorescence intensity were significantly higher (68.8 ± 5.3) compared with in the control group (45.0 ± 2.5). No differences were found in the levels of ROS or GSH between the treated and untreated groups. We conclude that hyaluronidase might trigger an increase in Ca2+ levels in oocytes, resulting in a decreased potential for normal embryonic development.

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Silica nanoparticles inducing the apoptosis of spermatocyte cell through microRNA-450b-3p targeting MTCH2-mediating mitochondrial signaling pathway

10.21203/rs.3.rs-24673/v1 ◽

2020 ◽

Author(s):

Guiqing Zhou ◽

Jianhui Liu ◽

Xiangyang Li ◽

Yujian Sang ◽

Yue Zhang ◽

...

Keyword(s):

Cytochrome C ◽

Silica Nanoparticles ◽

Caspase 3 ◽

Reproductive Toxicity ◽

Control Group ◽

Caspase 9 ◽

Protein Expressions ◽

Underlying Mechanisms

Abstract Background: Silica nanoparticles (SiNPs) are found in environmental particulate matter and are proven to have adverse effects on fertility. The relationship and underlying mechanisms between miRNAs and apoptosis induced by SiNPs during spermatogenesis is currently ambiguous. Experimental design: The present study was designed to investigate the role of miRNA-450b-3p in the reproductive toxicity caused by SiNPs. In vivo, 40 male mice were randomly divided into control and SiNPs groups, 20 per group. The mice in the SiNPs group were administrated 20 mg/kg SiNPs by tracheal perfusion once every 5 days, for 35 days, and the control group were given the equivalent of a normal luminal saline. In vitro, spermatocyte cells were divided into 0 and 5 μg/mL SiNPs groups, after passaged for 30 generations, the GC-2spd cells in 5 μg/mL SiNPs groups were transfected with miRNA-450b-3p and its mimic and inhibitor. Results: In vivo, the results showed that SiNPs damaged tissue structures of testis, decreased the quantity and quality of the sperm, reduced the expression of miR-450b-3p, and increased the protein expressions of the MTCH2, BID, BAX, Cytochrome C, Caspase-9, and Caspase-3 in the testis. In vitro, SiNPs obviously repressed the viability and increased the LDH level and apoptosis rate, decreased the levels of the miR-450b-3p, significantly enhanced the protein expressions of the MTCH2, BID, BAX, Cytochrome C, Caspase-9, Caspase-3; while the mimic of miR-450b-3p reversed the changes induced by SiNPs, but inhibitor further promoted the effects induced by SiNPs.Conclusion: The result suggested that SiNPs could induce the spermatocyte apoptosis by inhibiting the miR-450b-3p expression to target promoting the MTCH2 resulting in activating mitochondrial apoptotic signaling pathways in the spermatocyte cells.

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Antibacterial Activity of Lumbricus Rubellus Earthworm Extract Against Porphyromonas Gingivalis as the Bacterial Cause of Periodontitis

Open Access Macedonian Journal of Medical Sciences ◽

10.3889/oamjms.2019.222 ◽

2019 ◽

Vol 7 (6) ◽

pp. 1032-1036 ◽

Cited By ~ 2

Author(s):

I Gusti Agung Ayu Dharmawati ◽

Tjokorda Gde Bagus Mahadewa ◽

I Putu Eka Widyadharma

Keyword(s):

Antibacterial Activity ◽

Treatment Group ◽

Lumbricus Rubellus ◽

Control Group ◽

Inhibitory Zone ◽

Control Group Design ◽

Zone Diameter ◽

Significant Difference ◽

The Mean

AIM: The purpose of this study was to determine the antibacterial activity of Lumbricus rubellus earthworms through inhibitory zone diameter to the growth of the bacterium Phorphyromonas gingivalis as the cause of periodontitis. METHODS: This was an experimental study with randomised posttest-only control group design. The study was conducted at the Microbiology Research Center laboratory at the Faculty of Dentistry, Airlangga University, Indonesia. The study was conducted in vitro, the sample size was calculated using the Federer formula as many as four agar plates containing bacteria Phorphyromonas gingivalis, with each plate given five different treatments: control (ethanol), Lumbricus rubellus earthworm extract (ECT) with concentrations of 50%, 25%, 12.5%, and 6.25% respectively. The data in the form of inhibition zone diameter (measured in millimetres) obtained were tested using One-Way ANOVA. RESULTS: The mean diameter of the inhibitory zone extract of Lumbricus rubellus earthworm on the growth of Phorphyromonas gingivalis bacteria in the treatment group had significant differences (p < 0.05). The mean inhibition zones between controls and the ECT treatment group (ECT 50%, ECT 25%, ECT 12.5%) were statistically different (p < 0.05), in contrast with ECT 6.25% (p > 0.05) which did not show significant difference with the control group (p > 0.05). CONCLUSION: Lumbricus rubellus earthworm extract with a concentration of 50% has the largest diameter of the inhibitory zone on the growth of the Phorphyromonas gingivalis bacteria. The 6.25% earthworm extract showed no antibacterial activity against the growth of Phorphyromonas gingivalis bacteria.

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Transformasi Genetik Tanaman Kentang (Solanum tuberosum L.) dengan Gen acvB Menggunakan Vektor Agrobacterium tumefaciens

Agrotrop : Journal on Agriculture Science ◽

10.24843/ajoas.2021.v11.i01.p07 ◽

2021 ◽

Vol 11 (1) ◽

pp. 63

Author(s):

DARWIN SILALAHI ◽

I GEDE PUTU WIRAWAN ◽

MADE SRITAMIN

Keyword(s):

Solanum Tuberosum ◽

Agrobacterium Tumefaciens ◽

Treatment Group ◽

Genetic Transformation ◽

Solanum Tuberosum L ◽

Control Group ◽

Shoot Height ◽

Specific Pcr ◽

Convenient Technique

Agrobacterium tumefaciens Mediated Genetic Transformation of acvB Gene in Potato (Solanum tuberosum L.). Genetic transformations are now routinely applied to plant mediated by Agrobacterium tumefaciens as the most convenient technique. This study aimed to prove the success of A. tumefaciens mediated genetic transformation in potato. A. tumefaciens LBA (pBI 121) and explant of potato shoot were used in this study. Explants were grown in vitro on Murashige and Skoog media. Transformation was implemented using smear technique by smearing A. tumefaciens to injured explant. Experimental groups consisted of two groups: control group which did not receive transformation treatment and treatment group receiving transformation treatment. Explant growth was observed through the presence of shoots, branches and the shoot height. Explants in the treatment group resulted in a higher number of shoots, branches, and shoot heights compared to control. Phenol compounds appear in explant epidermal tissue, indicating the wounds produced by A. tumefaciens infection, thus the gene predicted to be transformed. Identification by PCR is needed to prove the existence of the acvB gene in potato plants genome, using acvB specific PCR primer as the marker, such as (5?-CCCT CTAG AGAC CCGC GCCA AGGCG-3?) and (5?CGCG TCGA CCTT GTCG GAAAG -3?) with 540-bp in base pair size produced.

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Effect of oocyte vitrification before and after in vitro maturation towards Bcl-2, Bax and Bcl-2/Bax ratio expression

Majalah Obstetri & Ginekologi ◽

10.20473/mog.v24i2.4559 ◽

2017 ◽

Vol 24 (2) ◽

pp. 56

Author(s):

Zakiyatul Faizah ◽

Haryanto Aswin ◽

Hamdani Lunardhi

Keyword(s):

Treatment Group ◽

Ethylene Glycol ◽

In Vitro Maturation ◽

Cumulus Cell ◽

Basic Medium ◽

Control Group ◽

Oocyte Vitrification ◽

Expression Ratio ◽

Before And After

Objectives: to compare the expression of Bcl-2, Bax and Bcl-2/Bax ratio in cumulus cell and oocyte between vitrified oocyte pre and post in vitro maturation.Materials and Methods: Maturation was operated in medium TC 100 µl for 24 hours. Vitrification begins with washing oocyte in PBS basic medium supplemented of 20% serum for 1-2 minutes, followed by equilibration medium PBS + 20% serum + 10% ethylene glycol for 10-14 minutes, then transferred to 20% serum + PBS + 0.5 M sucrose + 15% ethylene glycol + PROH 15% for 25-30 seconds. Thawing is processed by submerging the oocytes in the media: 1). PBS + 20% serum + 0.5 M sucrose, 2). PBS + 20% serum + 0.25 M sucrose, and 3). PBS + 20% serum + 0.1 M sucrose. Imunocytochemistry observed the expression of Bcl-2, bax and Bcl-2/bax ratio.Results: Bcl-2 expression on oocyte in control group differed significantly with treatment group, Bcl-2 expression on cumulus in control group differed significantly with treatment 1 group. Bax expression on oocyte in control group differed significantly with treatment group. Bax expression on cumulus in control group differed significantly with treatment group. Bcl-2/Bax expression ratio on oocyte and cumulus did not differ significantly in all groupConclusion: No difference Bcl-2/Bax expression ratio on oocyte and cumulus between vitrified oocyte pre and post in vitro maturation.

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L-carnitine Supplementation Enhances Nuclear and Cytoplasmic Maturation Rates of Sheep Oocytes In Vitro

Tropical Animal Science Journal ◽

10.5398/tasj.2021.44.2.131 ◽

2021 ◽

Vol 44 (2) ◽

pp. 131-137

Author(s):

Z. W. Bhakty ◽

E. M. Kaiin ◽

N. W. K. Karja ◽

M. A. Setiadi

Keyword(s):

Tissue Culture ◽

Treatment Group ◽

Culture Medium ◽

Fertilization Rate ◽

Control Group ◽

Carnitine Supplementation ◽

Progressive Motility ◽

Maturation Medium ◽

Cytoplasmic Maturation

The aim of the present study was to determine the effectiveness of l-carnitine (LC) supplementation on nuclear and cytoplasmic maturation rates of sheep oocytes. In experiment 1, oocytes were maturated for 24 hours in tissue culture medium 199 supplemented with LC at doses of 0.3 mg/mL, 0.6 mg/mL, and 0.9 mg/mL. In experiment 2, oocytes were maturated and fertilized in a media supplemented with LC at a dose of 0.3 mg/mL and incubated with 5x106 sperm/mL for 12 hours. The treatment group consisted of LC supplementation only in maturation medium (P1), only in fertilization medium (P2), and in both maturation and fertilization media (P3). In experiment 3, sperm motility patterns were assessed using CASA after being exposed to fertilization medium supplemented with LC at a dose of 0.3 mg/mL for 0 and 3 hours. Our results showed that supplementation of LC at a dose of 0.3 mg/mL significantly (p<0.05) increased the percentage of oocytes reaching metaphase II (86.7±4.1%) compared to those supplemented with LA at doses of 0, 0.6, and 0.9 mg/mL (73.6±1.2%, 81.4±1.3%, and 70.5±1.6%, respectively). The LC treatment in the fertilization medium only did not influence the number of two pronuclear formations (62.1±2.5%) compared to supplementation either in the maturation medium only (72.0±4.7%) or a combination of both in maturation and fertilization media (68.2±2.7%) (p<0.05). Further results after 3 hours of incubation compared to the control group showed the total motility (24.8±2.04% vs. 17.49±2.37%), progressive motility (14.17±2.03% vs. 6.49±1.64%), and curvilinear velocity (VCL) (119.70±3.73% vs. 71.15±10.59%) (p<0.05) were increased in the fertilization medium containing LC but it did not improve the fertilization rate. It is concluded that supplementation of LC at a dose of 0.3 mg/mL in the maturation medium only could better improve the nuclear and cytoplasmic maturation rates of sheep oocytes.

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Combination of Aerobics and Resistance Training to Obesity University Student Serum Visfatin Level Influence

The Open Biomedical Engineering Journal ◽

10.2174/1874120701509010424 ◽

2015 ◽

Vol 9 (1) ◽

pp. 424-428

Author(s):

Xuhui Li ◽

Xiaomei Fan

Keyword(s):

Resistance Training ◽

University Students ◽

Blood Lipid ◽

University Student ◽

Control Group ◽

Fat Percentage ◽

Before And After ◽

Group A ◽

Visfatin Level

Purpose: Discuss on combination of long-term aerobics and resistance training to obesity university student serum visfatin level and blood lipid metabolism influence, and reveal potential mechanism of losing weight through exercises. Method: Divide 30 obesity university students into 3 groups, control group (group C), aerobics group (group A as well as aerobics and resistance training combinative group (group A+R), take in 16 periods’ training, before and after intervention respectively test the three groups’ weight (W), body mass index (BMI),fat mass (FM), fat percentage (%F); serum TG,TCH,HDL-C,LDL-C content and visfatin level, result:16 after 16 weeks sports intervention, compare group A and group A+R with group C, W,BMI,FM,%F obvious drop (P<0.05); serum TG,TCH,LDL-C content obvious drop (P<0.05), serum HDL-C obvious rises (P<0.05), serum visfatin content obvious drops (P<0.05), compare group A+R with group A, the FM,%F and serum visfatin content obvious drop, other indicators have no significant differences. Relevant analysis finds that serum visfatin content is positive correlated to W, BMI, FM, F%, TG, TCH and LDL-C(P<0.05), and is negative correlated to HDL-C. Conclusion: Both 16 weeks’ aerobics as well as combination of aerobics and resistance training have good improvements to obesity university students’ weight and body composition, and have good adjustment on blood lipid metabolic disorder, and reduce serum visfatin level. Combination of aerobics and resistance training has more obvious impacts on FM, % F and serum visfatin content, and meanwhile, serum visfatin level is positive correlated to weight, BMI, %F, FM, TG, TCH, LDL-C, while is negative correlated to HDL-C.

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Effects of cigarette smoking on the growth of Streptococcus mutans biofilms: an in vitro study

10.21203/rs.3.rs-221658/v1 ◽

2021 ◽

Author(s):

Ye Han

Keyword(s):

Treatment Group ◽

Cigarette Smoking ◽

Streptococcus Mutans ◽

Laser Scanning ◽

Water Soluble ◽

Laser Scanning Microscopy ◽

Extracellular Polysaccharides ◽

Confocal Laser ◽

Control Group

Abstract This study aimed to investigate the differences in growth and virulence (EPSs and acidogenicity) of Streptococcus mutans biofilms (S. mutans) according to the different times of cigarette smoking (CS) treatment. S. mutans biofilms (74-hour-old) were formed on saliva-coated hydroxyapatite disks. The biofilms were treated with CS at different times per day (one time, three times, and six times/day). The control group did not receive CS treatment. Acidogenicity, dry weight, colony-forming units, water-soluble/insoluble extracellular polysaccharides, and intracellular polysaccharides were analyzed and confocal laser scanning microscopy images were obtained of the 74-h-old biofilms. The 74-h-old biofilms on sHA discs in the 6 times/day CS treatment group showed the lowest biofilm accumulation and extracellular polysaccharide amount compared with the control group and other CS treatment groups. In the CLSM study, the biofilms in the six times/day CS treatment group also showed the lowest bacterial count (live and dead cells) and EPS biovolume. CS has an obvious inhibition on the growth of S. mutans biofilms, the degree of inhibition is proportional to the number of CS treatments.

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Photodynamic Therapy Enhanced the Antitumor Effects of Berberine on HeLa Cells

Open Chemistry ◽

10.1515/chem-2019-0048 ◽

2019 ◽

Vol 17 (1) ◽

pp. 413-421 ◽

Cited By ~ 3

Author(s):

Han-Qing Liu ◽

Ya-Wen An ◽

A-Zhen Hu ◽

Ming-Hua Li ◽

Guang-Hui Cui

Keyword(s):

Photodynamic Therapy ◽

Western Blot ◽

Hela Cells ◽

Mammalian Target Of Rapamycin ◽

Control Group ◽

Antitumor Effects ◽

Underlying Mechanisms ◽

And Control

AbstractIn this study we investigated the antineoplastic effects of Berberine (BBR)-mediated photodynamic therapy (PDT) on HeLa cells and its related mechanisms. The CCK-8 assay and flow cytometry were used to evaluate the proliferation and apoptosis of cells respectively. In addition, changes in protein expression levels were assessed using western blot. BBR at dose of 10 mg/kg was injected intraperitoneally to mice with tumors and PDT treatments were performed 24 hours later. In vivo imaging systems were used to evaluate the fluorescence of BBR. In vitro, PDT significantly enhanced the effects of BBR on inducing cell apoptosis and inhibiting proliferation. The in vivo results showed that the fluorescence intensity in the PDT group was decreased compared with that in the BBR group. Tumor weights and tumor size in the PDT group were less than those in the control group; however, when BBR was applied without PDT, no significant differences were observed between the BBR and control group. The results of western blot showed that PDT enhanced the inhibitory effects of BBR on the mammalian target of rapamycin (mTOR) signaling pathway, that may partly explain the potential underlying mechanisms.

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EFFECTS OF ETHANOLIC EXTRACT OF RHINACANTHUS NASUTUS (L) KURZ AS AN ANTIMICROBIAL AGAINST ESCHERICHIA COLI BACTERIA USING IN VITRO METHOD

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2019.v12i12.35523 ◽

2019 ◽

pp. 69-72

Author(s):

SUNARTI M.BIOMED ◽

DEBORA PANINSARI

Keyword(s):

Escherichia Coli ◽

Treatment Group ◽

Ethanolic Extract ◽

Inhibition Zone ◽

Control Group ◽

Control Groups ◽

In Vitro Method ◽

E Coli ◽

Escherichia Coli Bacteria

Objective: The objective of this study was to discover of the ethanolic extract of Rhinacanthus nasutus (L) Kurz in inhibiting Escherichia coli bacteria using an in vitro method. Methods: This is an experimental study using a laboratory test with Kirby-Bauer or paper disc method by observing and measuring the inhibition zone of the ethanolic extract of R. nasutus against E. coli bacteria with extract concentrations of 15%, 30%, and 60% consisting of control groups and treatment group. The positive control group used chloramphenicol antibiotics and negative control groups used Aquadest. E. coli was incubated at 37°C for 24 h. Then, the plates were incubated for 24 h at 37°C and the diameter of the inhibition zone was observed until the 3rd day with three repetitions. Results: The results of the study showed that the mean inhibition zone of E. coli bacteria was 10.93 mm, 12.09 mm, and 18.90 mm. The results of the Shapiro–Wilk test were p=0.199. The results of the one-way analysis of variance test were p<0.05 and that of the post hoc test indicated a significant value of p<0.05. Based on the results of the research, there were significant differences in the inhibition zone between the control group and the treatment group at a concentration of 15%, 30%, and 60%. Conclusion: R. nasutus extract was effective to inhibit the growth of E. coli bacteria at concentrations of 15%, 30%, and 60%, so R. nasutus is effective as an antimicrobial.

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