27 Vitrification at the germinal vesicle stage does not trigger apoptosis in porcine oocytes and early embryos

2020 ◽  
Vol 32 (2) ◽  
pp. 139
Author(s):  
T. Somfai ◽  
H. T. Nguyen ◽  
M. T. Nguyen ◽  
T. Q. Dang-Nguyen ◽  
H. Kaneko ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Propylene Glycol ◽  
Science And Technology ◽  
Germinal Vesicle ◽  
Positive Control ◽  
Control Group ◽  
Mrna Levels ◽  
Cleavage Stage ◽  
Deoxyribonuclease I ◽  
Significant Difference

Porcine immature oocytes can survive vitrification at high rates and retain their ability to undergo maturation and fertilization; however, the procedure reduces their competence for subsequent embryo development via unknown mechanisms (Somfai et al. 2014 Plos One 9, e97731). The aim of the present study was to clarify whether our vitrification procedure at the germinal vesicle stage triggers apoptosis in oocytes and subsequent developing embryos. Immature porcine cumulus-oocyte complexes obtained from slaughterhouse-derived ovaries were vitrified and warmed by our method (Appeltant et al. 2018 Cryobiology 85, 87-94) immediately after collection (vitrified group). The oocytes were equilibrated in 2% (vol/vol) ethylene glycol and 2% (vol/vol) propylene glycol for 13-15min. Then, they were vitrified by dropping them into liquid nitrogen in 2-μL microdrops of a medium composed of 17.5% ethylene glycol, 17.5% propylene glycol, 0.3M sucrose, and 50mgmL−1 polyvinylpyrrolidone. After warming, they were subjected to IVM, fertilization (IVF), and embryo culture using chemically defined media (Yoshioka et al. 2008 J. Reprod. Dev. 54, 208-213). From each collected batch, a group of oocytes was processed without vitrification (control group). Apoptosis was assayed in membrane-intact oocytes at the end of IVM and in cleavage-stage embryos on Day 2 after IVF (Day 0) by the CaspACE FITC-VAD-FMK In Situ Caspase Marker (Promega; Experiment 1), deoxynucleotidyl transferase (TdT) dUTP nick-end labelling (TUNEL; Experiment 2), and analysis of mRNA levels by RT-qPCR for the pro-apoptotic Bax and CASP3 genes (Experiment 3). Each experiment was replicated three times. Data were analysed by Kruskal-Wallis test followed by Dunn's multiple comparisons test. The mean survival rate of vitrified oocytes was 89.2%. There was no significant difference between the control and vitrified groups in relative caspase levels in IVM oocytes and in 2- to 4-cell embryos after IVF; however, significantly increased caspase activity (P<0.05) was detected in oocytes and embryos after treatment with 10 μM staurosporine (positive control). There was no significant difference between the control and vitrified groups in the proportion of TUNEL-positive oocytes (4.1 and 0.8%, respectively) and embryos (0 and 0%, respectively), whereas 96.6% of oocytes and 100% of cleavage stage embryos treated with 1000IUmL−1 deoxyribonuclease I (positive control) were proven to be TUNEL positive (P<0.05). Similar mRNA levels for Bax and CASP3 genes were detected in oocytes at the end of IVM and subsequent developing 4- to 8-cell embryos between the control and vitrified groups. In conclusion, vitrification of porcine oocytes at the germinal vesicle stage by our method did not trigger apoptosis in oocytes and subsequent developing embryos. This work was supported by the Japan Science and Technology Agency (JST)/Japan International Cooperation Agency (JICA) Science and Technology Research Partnership for Sustainable Development (SATREPS).

Potential Antiseptic of Rhodomyrtus tomentosa leaves extract on Healing wound in Male Wistar rats

2021 ◽  
pp. 3143-3149
Author(s):  
Endang Sri Purwanti Ningsih ◽  
Noorlaila Noorlaila ◽  
Ikhwan Rizki Muhammad ◽  
Windy Yuliana Budianto
Keyword(s):  
Wound Healing ◽  
Wistar Rats ◽  
Leaf Extract ◽  
Wound Care ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Significant Difference ◽  
Male Wistar Rats ◽  
Rhodomyrtus Tomentosa

Background: The process of wound healing is influenced by various factors such as age, hormones, and wound care. Wound care is done to accelerate wound healing which can be done by various methods, one of them is traditional care. Traditional wound care can use medicinal plants. Rhodomyrtus tomentosa is a medicinal plant that has an antioxidant, anti-inflammatory, antitumor and antibacterial content. Thus this study aims to evaluate the effectiveness of the antiseptic solution of the Rodhomyrtus tomentosa leaf extract on wound healing in male Wistar rats. Method: this research is pure experimental research with post test only control group design. Thirty male white rats were divided into five groups, namely negative control, positive control, Rhodomyrtus tomentosa leaf extract 15%, 30%, and 60%. Rhodomyrtus tomentosa leaf extraction was carried out by maceration method with 70% ethano solvent. The extraction results are divided into 3 concentrations (15%, 30% and 60%). The wound healing process was evaluated by measuring the length of the wound manually from 0 to 10 days in each group. Meanwhile, the number of fibroblast cells was calculated through hematoxylin eosin (HE) staining and observed using an Olympus CX41 microscope with a 10x magnification and objective lens magnification in 3 fields. Result: There was a significant difference in the reduction in wound length (p =< 0,000) between the five experimental groups (Rhodomyrtus tomentosa leaf extract solution 15%, 30% and 60%, negative control and positive control. Solution of rhodomyrtus tomentosa leaf extract accelerated the increase in the number of fibroblasts compared to the negative control group (p = 0.003), but did not make a difference (p = 0.403) with the positive control group. Rhodomyrtus tomentosa leaf extraction solution had the same microscopic effect on the number of fibroblasts with a positive control group given 0.9% NaCl solution. Conclusion: There was a significant difference in the number of fibroblasts between all groups, but no difference in wound healing length.

scholarly journals Coenzyme q10 is potentially usedto improve lipid profile in diabetic hypercholesterolemia-induced periodontitis in the coastal area

2018 ◽  
Vol 47 ◽  
pp. 07006
Author(s):  
Saryono Saryono ◽  
Hesti Devinta ◽  
Abdul Haris Budi Widodo ◽  
Arif Imam Hidayat
Keyword(s):  
Lipid Profile ◽  
Coenzyme Q10 ◽  
Coastal Area ◽  
Positive Control ◽  
Atherogenic Index ◽  
Control Group ◽  
Treatment Groups ◽  
Significant Difference ◽  
Pap Method ◽  
Post Hoc

Contaminated water often affects the occurrence of periodontitis in the coastal area. The diabetic hypercholesterolemia-induced periodontitis in Indonesia is very high. The use of coenzyme Q10 to treat this disease has never been investigated yet. Therefore, this study aimedto analyze the effect of coenzyme Q10 on the lipid profile of diabetic hypercholesterolemia-induced periodontitis.Twenty four rats were randomized into 6 groups (G1-G6). The groups (G1-G3) are healthy, negative and positive control group respectively. The treatment groups (G4-G6) are diabetic hypercholesterolemia-induced rats given coenzyme q10 dose of 13,5; 27 and 54 mg/kg respectively. Rats were induced by periodontitis, hypercholesterolemia and diabetes mellitus. Coenzyme Q10 was administered orally using 2 mL gastric tube once a day for 14 days. Lipid profile including triglycerides, HDL, and atherogenic index (IA) was measured enzymatically by the CHOD-PAP method. Data were analyzed by one-way ANOVA test and followed by the Least Significant Difference (LSD) post hoc test.Coenzyme Q10 with a dose of 54 mg/kgis effective in lowering triglyceride, and atherogenic index and increasing HDL level in diabetic hyperlipidemia-induced periodontitis rats model. This research supports the potential effects of coenzyme Q10 supplementation to improve lipid profile in diabetic hypercholesterolemia-induced periodontitis in the coastal area.

scholarly journals Evaluation of the Sealing Ability of Three Obturation Techniques Using a Glucose Leakage Test

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Katarzyna Olczak ◽  
Halina Pawlicka
Keyword(s):  
Continuous Wave ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Thermal Methods ◽  
Root Canals ◽  
Anterior Teeth ◽  
Sealing Ability ◽  
Significant Difference

The aim of this study was to evaluate the sealing ability of three different canal filling techniques. Sixty-four roots of extracted human maxillary anterior teeth were prepared using ProTaper® rotary instruments. The specimens were then randomly divided into 3 experimental groups (n=16) and 2 control groups (n=8). The root canals were filled using cold lateral compaction (CLC group), continuous wave condensation technique using the Elements Obturation Unit® (EOU group), and ProTaper obturators (PT group). For the negative control group, 8 roots were filled using lateral compaction as in the CLC group, and the teeth were covered twice with a layer of nail varnish (NCG group). Another 8 roots were filled using lateral compaction, but without sealer, and these were used as the positive control (PCG group). A glucose leakage model was used for quantitative evaluation of microleakage for 24 hours and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 weeks. No significant difference in the cumulative amount of leakage was found between the three experimental groups at all observation times. The lateral condensation of cold gutta-percha can guarantee a similar seal of canal fillings as can be achieved by using thermal methods, in the round canals.

Human oocyte vitrification with corona radiata, in autologous follicular fluid supplemented with ethylene glycol, preserves conventional IVF potential: birth of four healthy babies

2014 ◽  
Vol 26 (7) ◽  
pp. 1001 ◽  
Author(s):  
Xian-Hong Tong ◽  
Li-Min Wu ◽  
Ren-Tao Jin ◽  
Hong-Bing Luan ◽  
Yu-Sheng Liu
Keyword(s):  
Ethylene Glycol ◽  
Embryo Transfer ◽  
Follicular Fluid ◽  
Control Group ◽  
Human Oocyte ◽  
Oocyte Vitrification ◽  
Human Oocytes ◽  
Corona Radiata ◽  
Fertilisation Rate ◽  
Significant Difference

The aim of this study was to examine the effects of vitrification with autologous follicular fluid (AFF) supplemented with ethylene glycol (EG) and sucrose on human oocytes with corona radiata. A total of 182 human oocytes with corona radiata from fifteen infertile patients were vitrified using either equilibration solutions (ES) and vitrification solution (VS) consisting of AFF, EG and sucrose (AFF group, n = 67) or commercial ES and VS (control group, n = 115). All oocytes were thawed in the next cycle, with surviving oocytes being inseminated by conventional IVF. The clinical outcome of vitrified–warmed oocytes by both vitrification methods was analysed retrospectively. In the AFF group, six patients received embryo transfer, with three couples taking four healthy babies home. In the control group, nine patients received embryo transfer, with four couples taking five healthy babies home. There was no significant difference in the survival rate (91.0 vs 92.2%), two pronuclei (2PN) fertilisation rate (73.8 vs 73.6%), cleavage rate (100 vs 100%), top-quality embryo rate (62.2 vs 59.2%), clinical pregnancy rate (50.0 vs 44.4%), implantation rate (33.3 vs 25%) or take-home baby rate (50.0 vs 44.4%) between the AFF group and the control group, respectively. These results show that AFF supplemented with EG and sucrose is an efficient, cost-effective cryoprotectant for human oocyte cryopreservation. A corona radiata on vitrified–warmed oocytes retains the oocytes’ fertilisation capability in conventional IVF.

scholarly journals Screening of natural product extracts for fly repellent and larvicide

2021 ◽  
Vol 20 (1) ◽  
pp. 25-40
Author(s):  
Kanokwan Bootyothee ◽  
◽  
Surasit Aunpromma ◽  
Atchara Artchayasawat ◽  
Opal Pitaksakulrat ◽  
...  
Keyword(s):  
Natural Products ◽  
Mortality Rate ◽  
Crude Extract ◽  
Rural Areas ◽  
Inorganic Compounds ◽  
Food Contamination ◽  
Positive Control ◽  
Positive Control Group ◽  
Control Group ◽  
Significant Difference

Fly is one of the vectors of foodborne pathogenic vectors and causes myiasis in humans and animals. To prevent the contamination of food and myiasis, various chemical products are commonly used. However, eco-friendly is now a trend all over the world so natural products are the alternative ways to reduce the chemical residue in the environment. The present study was screened the 12 natural products in 4 groups; the essential oil plants, the alkaloid plants, the cyanogenic plant, and the inorganic compounds which use a simple extraction on the fly repelling and larvicide. The fly repelling efficacy with 2 criteria consist of the time of the first swarm and percent repelling (PR) in the first 15 min. The larvicidal efficacy was determined by the mortality rate within 24 h. The Kaffir lime peels aqueous crude extract exhibited the best result on fly repelling from both criteria with no significant difference. The bamboo shoots aqueous crude extract gave the best result on larvicide with significantly different from other groups (P < 0.001) which was faster than the positive control group (Coumaphos & Propoxur). Moreover, the high mortality rate reached 73.33 ± 15.28% within 24 h, higher than the positive control group. The present study indicates the power of common natural products on fly repellent and larvicide that may apply for fly repellant to reduce the food contamination especially fresh materials and prevention or treatment of animal and human myiasis in rural areas

scholarly journals A Digital Microscopic Inspection of Dentinal Defects after Using Endodontic Retreatment Files

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Ammar AbuMostafa ◽  
Hala Almoqayyad ◽  
Al-Omari Mohammad
Keyword(s):  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Filling Material ◽  
Root Canals ◽  
Root Canal Retreatment ◽  
Significant Difference ◽  
Digital Microscope ◽  
Microscopic Inspection

Aim. The study aimed at evaluating the incidence of dentinal defects after root canal retreatment with ProTaper Universal retreatment (PTUR) and XP-endo Shaper and Finisher R (XP). Materials and Methods. Sixty extracted single-rooted human premolars were selected and divided into 4 groups of 15 teeth each. In the negative control group, the teeth were left unprepared. In the positive control group, the teeth were prepared with ProTaper Next and obturated with no further retreatment. In the PTUR and XP groups, the teeth were prepared and obturated followed by removal of the filling material at body temperature using PTUR and XP instruments, respectively. The roots were then sectioned at 3, 6, and 9 mm from the apex and observed under a digital microscope to detect defects. Results. PTUR group showed significantly higher ( p value <0.05) incidence of defects than the other groups. Comparison of no defects versus defects between groups in different areas of root canals demonstrated significant difference among the groups in the apical and cervical regions. Conclusion. Within the limitations of the present study, PTUR files created significantly more dentinal defects than XP files, with most of those defects at the cervical and apical areas of the root canals.

Comparison of Biocompatibility of Experimental Tricalcium Phosphate Cement versus Biodentin and Mineral Trioxide Aggregate used for Furcation Perforation Repair (in vivo study)

2021 ◽  
Vol 15 (1) ◽  
pp. 532-538
Author(s):  
Mostafa M. El-Bialy ◽  
Magdy M. Ali ◽  
Engy M. Kataia ◽  
Reham S. El Nemr ◽  
Reham Hassan
Keyword(s):  
Tricalcium Phosphate ◽  
Mineral Trioxide Aggregate ◽  
Positive Control ◽  
Control Group ◽  
Tooth Survival ◽  
Significant Difference ◽  
Operative Evaluation ◽  
Root Canal System ◽  
Bone Deposition

Background: The interaction between the root canal system and the oral cavity caused by iatrogenic perforations significantly affects the treatment outcome and tooth survival. Objectives: This study was directed to compare the biocompatibility of an experimental tricalcium phosphate cement versus biodentine and mineral Trioxide aggregate used for furcation perforation repair in dogs. Methods: Perforations were done in 60 teeth (premolars and molars) of six adult dogs. Animals were divided randomly into 3 equal groups of 2 animals each according to the post-operative evaluation period of 1 week, 1 month, and 3-months. Each group was further subdivided into 4 subgroups according to either being repaired with the experimental tricalcium phosphate cement (n=6), or Biodentine (n=6), Mineral Trioxide Aggregate (MTA) (n=6), and positive control(n=2). After evaluation periods, tissue blocks were harvested and histologically examined. Results: No statistically significant difference was found regarding bone deposition scores and inflammatory reaction in the three groups after 1 week, 1 month, or 3 months. All three groups showed a statistically significant difference between all three time periods. Regardless of the repair material used, inflammation scores at 1-week showed the highest scores, decreasing over time except for the control group. Conclusion: The experimental material could be considered as a successful treatment option for repairing furcation perforation.

25 Effect of taxol and epothilone B on meiotic spindle stabilization in vitrified bovine oocytes

2021 ◽  
Vol 33 (2) ◽  
pp. 120
Author(s):  
E. Girka ◽  
K. R. Bondioli
Keyword(s):  
Ethylene Glycol ◽  
Dimethyl Sulfoxide ◽  
Recovery Period ◽  
Normal Chromosome ◽  
Meiotic Spindle ◽  
Control Group ◽  
Chromosome Arrangement ◽  
Epothilone B ◽  
Significant Difference ◽  
Bovine Oocytes

Vitrification has the potential to be a valuable technique for preservation of bovine oocytes; however, this method often results in abnormal microtubule and chromosome arrangement. The aim of this experiment was to evaluate taxol and epothilone B as meiotic spindle stabilising pretreatments in a vitrification protocol. Bovine oocytes were purchased and matured invitro during shipment. At 18h of maturation, oocytes were divided randomly into control, taxol, and epothilone B treatments (Table 1). All treatments were prepared in invitro maturation (IVM) medium (IVF Biosciences). Partially denuded oocytes were incubated in either control or treatment medium for 15min at 38.5°C before vitrification. Oocytes were incubated in an equilibration solution (10% dimethyl sulfoxide, 10% ethylene glycol) for 5min, transferred to a vitrification solution (20% dimethyl sulfoxide, 20% ethylene glycol, 0.5M sucrose), loaded onto a Cryolock, and plunged into liquid nitrogen within 45s. For warming, a Cryolock was placed directly into a 0.5M sucrose solution and incubated for 3min. Oocytes were transferred to a 0.25M solution for 3min and washed in the basal solution used for vitrification and warming media (Dulbecco’s phosphate-buffered saline, 20% fetal bovine serum). Once warmed, oocytes were transferred to IVM medium for a 4-h recovery period and completely denuded before staining. Staining to evaluate spindle morphology was performed with anti α-tubulin primary antibody and secondary antibody Alexa Fluor 488. Oocytes were also stained with Hoechst to evaluate chromosome arrangement. Both spindle morphology and chromosome arrangement data were analysed using a logistic regression with a binomial response variable (normal/abnormal). Both 0.5μM and 1.0μM Taxol treatments had no effect on either meiotic spindle or chromosome arrangement compared with the control group (P&gt;0.05). The 2.0μM taxol treatment improved chromosome configuration (P&lt;0.05) with no effect on microtubule distribution compared with the control group (P&gt;0.05). All epothilone B treatments resulted in disruption of microtubule distribution and chromosome arrangement compared with control (P&lt;0.001) and resulted in a consistent abnormality hypothesised to be tubulin polymerization. These results indicate that taxol is capable of increasing the occurrence of normal chromosome arrangement in vitrified bovine oocytes and that epothilone B may cause additional harm to the oocyte that is not associated with the metaphase plate. Table 1. Effect of stabilisation agents on meiotic spindle of invitro-matured bovine oocytes Treatment n Normal microtubule distribution (%) Normal chromosome arrangement (%) Control 100 44 47 0.5μM Taxol 104 44 37 1.0μM Taxol 98 43 56 2.0μM Taxol 102 49 62a 0.5μM Epothilone B 103 11b 11b 1.0μM Epothilone B 97 6b 8b 2.0μM Epothilone B 100 2b 1b aP&lt;0.05;. bP&lt;0.001: Different superscripts within a column indicate a significant difference.

scholarly journals HISTOPATHOLOGICAL STRUCTURE OF WHITE RATS LIVER AFTER GIVING ANT NEST EXTRACT DUE TO INDUCED PARACETAMOL TOXIC DOSE

2019 ◽  
pp. 14
Author(s):  
I Made Merdana ◽  
I Made Kardena ◽  
Ketut Budiasa ◽  
I Made Dodi Gunawan
Keyword(s):  
Rat Liver ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Toxic Dose ◽  
White Rats ◽  
Significant Difference ◽  
Ant Nest ◽  
Histopathologic Changes

This study aim was to determine the influence ant nest plant extract (Myrmecodia pendans) on histopathological changeof white rat liver (Rattus novergicus) due to induced with paracetamol toxic dose. This study used 24 male white rats, divided into four groups, negative control group (P0) given placebo, positive control group (P1) given paracetamol dose 250 mg / kg bw for 10 days, P2 given ant nest extract 250 mg / kg bw and paracetamol dose 250 mg / kg bw for 10 days, P3 given ants nest extract 250 mg / kg bw for seven days, then continued by giving paracetamol and ants nest extract with dose 250 mg / kg bw for ten days. After the treatment done, all the rats were dinecropsed. Liver organs were taken and processed for making histopathology preparations. Parameters examined included hemorrhage, congestion, degeneration and necrosis. The data obtained were analyzed statistically by using Kruskal Wallis test followed by Mann Whitney test. Mann Whitney test results for all categories of histopathologic changes in  hemorrhagic, congestion, degeneration, and necrosis between negative control group (P0) and positive control group (P1) were significantly different (P <0.05), between negative control (P0) with P2 and P3 there was no significant difference (P> 0,05). Afterward, between the positive control (P1) and P2 with P3 there was a significant difference (P <0.05). I can be concludedthat the administration of paracetamol dose 250 mg/kg bw for 10 days affects the histopathologic changes of white rat liver. The administration of ant nest plant extracts can reduce the side effects of toxic doses of paracetamol.

scholarly journals Efficacy of a herbal toothpaste on patients with established gingivitis: a randomized controlled trial

2006 ◽  
Vol 20 (2) ◽  
pp. 172-177 ◽  
Author(s):  
Fabiana Ozaki ◽  
Claudio Mendes Pannuti ◽  
Ana Vitória Imbronito ◽  
Wellington Pessotti ◽  
Luciana Saraiva ◽  
...  
Keyword(s):  
Adverse Reactions ◽  
Controlled Trial ◽  
Test Group ◽  
Experimental Period ◽  
Positive Control ◽  
Control Group ◽  
Control Groups ◽  
Double Blind ◽  
Significant Difference ◽  
And Control

The aim of this randomised, double blind controlled trial was to verify the efficacy of a herbal dentifrice on the reduction of plaque and gingivitis. Forty eight volunteers with established gingivitis were randomly assigned to either a test group (herbal dentifrice) or positive control group (dentifrice with triclosan and fluoride). The dentifrices were distributed in plain white tubes by an independent pharmacy, which revealed the contents of each tube only after the experimental period. Plaque and gingivitis assessments were carried out on baseline and after 28 days of product use. All examinations were conducted by the same calibrated investigator. Subjects were instructed to brush their teeth three times daily using their assigned dentifrice for 28 days. There was a significant reduction in plaque levels in both the test and control groups. However, there was no significant difference between the groups. A significant reduction in gingivitis was observed in both groups, although there was no significant difference between them. No adverse reactions were reported. The authors concluded that both dentifrices were effective in reducing plaque and gingivitis in subjects with established gingivitis.

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