Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro

Zygote ◽  
2020 ◽  
Vol 28 (6) ◽  
pp. 504-510
Author(s):  
P.A.A. Barroso ◽  
L. R F.M. Paulino ◽  
B.R. Silva ◽  
G.L. Vasconcelos ◽  
D.S. Gomes ◽  
...  
Keyword(s):  
Analysis Of Variance ◽  
Granulosa Cells ◽  
Ultrastructural Analysis ◽  
Culture Period ◽  
Cavity Formation ◽  
Significant Difference ◽  
Chi Squared ◽  
Calcein Am

SummaryThis study aimed to evaluate the effects of dexamethasone on development, viability, antrum formation and ultrastructural integrity of bovine secondary follicles cultured in vitro for 18 days. Bovine ovaries were obtained from slaughterhouses and secondary follicles of ~150–200 µm diameter were isolated and cultured in the laboratory in TCM-199+ alone or supplemented with different concentrations of dexamethasone (1, 10, 100 and 1000 ng/ml). Follicle viability was evaluated after the culture period, using calcein-AM (viable) and ethidium homodimer (nonviable). Follicle diameters and antrum formation were evaluated at days 0, 6, 12 and 18. Before or after in vitro culture, follicles were fixed for histological and ultrastructural analysis. Follicle diameters were evaluated using analysis of variance and Kruskal–Wallis test, while chi-squared test was used to evaluate the percentage of viable follicles and antrum formation (P < 0.05). Follicles cultured for 6 days with all treatments increased their diameters significantly, but there was no significant difference between treatments at the end of the culture period. In vitro cultured follicles showed antral cavity formation at the end of the culture period, but no influence of dexamethasone was seen. Ultrastructural analysis showed that follicles cultured with dexamethasone (1, 10, 100 and 1000 ng/ml) had well preserved granulosa cells. However, oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone showed signs of degeneration. It can be concluded that follicles cultured in vitro in the presence of dexamethasone demonstrated continuous in vitro growth, but oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone had poor ultrastructure.

scholarly journals KECERNAAN IN VITRO SILASE SAMPAH SAYUR DAN DAUN GAMAL MENGGUNAKAN MIKROORGANISME RUMEN KAMBING

2015 ◽  
Vol 2 (3) ◽  
pp. 17
Author(s):  
Indrayani Indrayani ◽  
Harapin Hafid ◽  
Dian Agustina
Keyword(s):  
Organic Matter ◽  
Analysis Of Variance ◽  
Dry Matter ◽  
Vegetable Waste ◽  
Dry Matter Digestibility ◽  
Randomized Design ◽  
Honestly Significant Difference ◽  
Significant Difference ◽  
Completely Randomized Design

ABSTRACTThis study aims to determine the digestibility of dry matter and organic matter level waste silage mixed vegetables and Gliricidia leaves were tested in vitro. This study used a completely randomized design with 3 treatments and 3 replications. Each of these treatments is R0 (Gliricidia leaves 100%), R1 (Gliricidia leaves 70% + 30% silage vegetable waste), and R2 (Gliricidia leaves 40% + 60% silage vegetable waste). Data were analyzed using analysis of variance (ANOVA) and further testing using the test Honestly Significant Difference (HSD). The results of ANOVA showed that the mixture of vegetable waste silage was highly significant (p <0.05) on dry matter digestibility and significantly (p <0.05) on the digestibility of organic matter. It can be concluded that the mixed of vegetable waste silage and Gliricidia leaves can improved digestibility of dry matter and organic matter, treatment of 40% and 60% Gliricidia leaves plus waste vegetable produce silage dry matter digestibility and percentage of organic matter is best (72,24% and 68,19%).Keyword: Silage vegetable waste, gliricidia leaves, dry and organic matter digestibilityABSTRAKPenelitian ini bertujuan untuk mengetahui kecenaan bahan kering dan bahan organik tingkat campuran silase sampah sayur dan daun gamal yang diuji secara in vitro. Penelitian ini menggunakan Rancangan Acak Lengkap dengan 3 perlakuan dan 3 ulangan. Masing-masing perlakuan tersebut ialah R0 (daun gamal 100%), R1 (daun gamal 70% + silase sampah sayur 30%), dan R2 (daun gamal 40% + silase sampah sayur 60%). Data yang diperoleh dianalisis dengan menggunakan analisis sidik ragam (ANOVA) dan uji lanjut menggunakan uji Beda Nyata Jujur (BNJ). Hasil analisis sidik ragam menunjukkan bahwa campuran silase sampah sayur berpengaruh sangat nyata (p<0,05) terhadap kecernaan bahan kering dan berpengaruh nyata (p<0,05) terhadap kecernaan bahan organik. Dapat disimpulkan bahwa perlakuan campuran silase sampah sayur dan daun gamal dapat meningkatkan kecernaan bahan kering dan bahan organik, perlakuan 40% daun gamal dan 60% silase sampah sayur menghasilkan persentase kecernaan bahan kering dan bahan organik yang terbaik yaitu (72,24% dan 68,19%).Kata kunci : Silase sampah sayur, daun gamal, kecernaan bahan kering, dan bahan organik.

Performance Characteristics and Accuracy in Perceptual Discrimination of Leather and Synthetic Basketballs

1982 ◽  
Vol 55 (1) ◽  
pp. 128-130 ◽  
Author(s):  
Sharon Mathes ◽  
Kay Flatten
Keyword(s):  
Analysis Of Variance ◽  
Performance Characteristics ◽  
Perceptual Discrimination ◽  
Treatment Conditions ◽  
Significant Difference ◽  
Chi Squared ◽  
Perceptual Mode

Performance characteristics of leather and synthetic basketballs were examined by measuring the basketballs' rebound heights on five types of playing surfaces (Tartan, asphalt, glass, concrete, hardwood). Comparing the basketballs' performance on the basis of their coefficients of restitution (e = height of rebound/height of drop) analysis of variance showed that the leather basketball rebounded significantly higher than the synthetic basketball on all surfaces. To assess the capability of individuals to discriminate perceptually between the balls 30 male and 30 female undergraduates were asked to determine whether basketballs randomly presented 20 times under four different treatment conditions (visual, tactual-kinesthetic static, tactual-kinesthetic dynamic, auditory) were leather or synthetic. Chi squared analysis of their accuracy across all four perceptual modes showed no significant difference. However, analysis by perceptual mode did produce significant differences, indicating subjects were more accurate in identification in the tactual-kinesthetic dynamic and static modes than in the visual and auditory.

scholarly journals Apical Extrusion of Intracanal Debris Using Two Engine Driven and Step-Back Instrumentation Techniques: An In-Vitro Study

2008 ◽  
Vol 02 (04) ◽  
pp. 233-239 ◽  
Author(s):  
Alper Kustarci ◽  
Neslihan Akdemir ◽  
Seyda Herguner Siso ◽  
Demet Altunbas
Keyword(s):  
Stainless Steel ◽  
Analysis Of Variance ◽  
Statistical Significance ◽  
In Vitro Study ◽  
Type Stainless Steel ◽  
Rotary Instruments ◽  
Apical Extrusion ◽  
Working Length ◽  
Significant Difference

ABSTRACTObjectives: The purpose of this study was to compare in-vitro the amount of debris extruded apically from extracted teeth, using K3, Protaper rotary instruments and manual step-back technique.Methods: Forty five human single-rooted mandibular premolar teeth were randomly divided into 3 groups. The teeth in 3 groups were instrumented until reaching the working length with K3, Protaper rotary instruments and K-type stainless steel instruments with manual step-back technique, respectively. Debris extruded from the apical foramen was collected into centrifuge tubes and the amount was determined. The data obtained were analyzed using Kruskal-Wallis one-way analysis of variance and Mann-Whitney U tests, with P=.05 as the level for statistical significance.Results: Statistically significant difference was observed between K3, Protaper and step-back groups in terms of debris extrusion (P<.05). Step-back group had the highest mean debris weight, which was significantly different from the K3 and Protaper groups (P<.05). The lowest mean debris weight was related to K3 group, which was significantly different from the Protaper group (P<.05).Conclusions: Based on the results, all instrumentation techniques produced debris extrusion. The engine-driven Ni-Ti systems extruded significantly less apical debris than step-back technique. However, Protaper rotary instruments extruded significantly more debris than K3 rotary instruments. (Eur J Dent 2008;2:233-239)

31 PROTOCOL OPTIMIZATION AND EVALUATION OF MATURATION PROMOTING FACTOR AND MITOGEN-ACTIVATED PROTEIN KINASE ACTIVITIES IN BOVINE CYTOPLASTS OBTAINED BY CHEMICAL ENUCLEATION TECHNIQUES

2014 ◽  
Vol 26 (1) ◽  
pp. 130
Author(s):  
N. Z. Saraiva ◽  
C. S. Oliveira ◽  
M. del Collado ◽  
M. R. de Lima ◽  
R. Vantini ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Mitogen Activated Protein Kinase ◽  
Mapk Activity ◽  
Significant Difference ◽  
Mitogen Activated Protein ◽  
Chi Squared ◽  
High Concentrations ◽  
Bovine Oocytes ◽  
Maturation Promoting Factor

Chemical enucleation using microtubule-depolymerizing drugs is an attractive procedure to simplify the enucleation process in nuclear transfer. The aim of this study was to optimize chemically assisted (CA) and chemically induced (CI) enucleation protocols using metaphase II (MII) and pre-activated bovine oocytes, respectively, and to evaluate the activity of maturation promoting factor (MPF) and mitogen-activated protein kinase (MAPK) in cytoplasts generated by these techniques. Initially, we determined the shortest effective treatment of MII and activated oocytes with 0.05 μg mL–1 demecolcine. Bovine oocytes in vitro matured (IVM) for 19 h (MII) or activated artificially with 5 μM ionomycin (5 min) and 10 μg mL–1 cycloheximide (5 h) after 26 h IVM were treated with demecolcine and samples were collected at 0, 0.25, 0.5, 1.0, 1.5, and 2.0 h of treatment. Oocytes were then stained with 10 μg mL–1 Hoechst 33342 and the protrusion or enucleation rates were determined. Next, we evaluated histone H1 and myelin basic protein (MBP) kinases, reflecting MPF and MAPK activities, respectively, in oocytes obtained from these treatments, and for that we used the method described by Kubelka et al. (2000 Biol. Reprod. 62, 292–302). Protrusion and enucleation rates were evaluated by the chi-squared (χ2) test, and MPF and MAPK activities were submitted to ANOVA and Tukey's test at 5% significance. For MII oocytes, effects of demecolcine were observed as early as 15 min, with a significant difference (P < 0.05) between control (12/112, 10.7%) and treated (33/114, 28.9%) groups in relation to protrusion rates. The largest number of protrusions was observed after 1.0 h of treatment (control: 15/113, 13.3%a; treated: 45/111, 40.5%b). In pre-activated oocytes, effects of demecolcine were also observed after 15 min, and in both techniques there were no significant differences between groups treated with demecolcine for 1.0, 1.5, or 2.0 h (CA: 40.5 to 52.5% of protrusion; CI: 35.2 to 46.7% of enucleation). In contrast to previous reports in which high concentrations of demecolcine for CA enucleation increased MPF activity, we observed no alterations in the activity of this factor at a demecolcine concentration of 0.05 μg mL–1. Activity of MAPK also did not differ significantly between the control and treated groups throughout evaluation. In the CI technique, a significant difference in MPF activity was observed after 0.5 h (70.3%) and 2.0 h of activation (39.1%), considering that the activity was 100% at the beginning of the evaluation. However, we observed no significant difference between the control and treated groups at any of the time points studied, as verified for MAPK activity. The exact effect of MPF on the nucleus in mammals is not well established. We believe that the use of low concentrations of demecolcine for short periods is less damaging to embryonic development and, until we have a better understanding of the effect of these kinases on the transferred nucleus, we recommend its use for chemical enucleation protocols in bovine. Financial support: FAPESP 2010/20744-6 and 2011/12983-3.

334. PROGESTERONE PRODUCTION FROM GRANULOSA CELLS OF SOWS IS ENHANCED EQUALLY BY OMEGA-3 DERIVED PROSTAGLANDIN E3 AND OMEGA-6 DERIVED PROSTAGLANDIN E2

2010 ◽  
Vol 22 (9) ◽  
pp. 134
Author(s):  
R. Smits ◽  
D. T. Armstrong ◽  
L. Ritter ◽  
M. Mitchell ◽  
M. B. Nottle
Keyword(s):  
Prostaglandin E2 ◽  
Granulosa Cells ◽  
Luteinizing Hormone Receptor ◽  
Follicle Cells ◽  
Omega 3 ◽  
Progesterone Production ◽  
Porcine Granulosa Cells ◽  
Significant Difference ◽  
Omega 6

Caughey et al (2005) reported that prostaglandins derived from omega 3 sources eicsapentaenoic acid (EPA C20:5) and docosahexaenoic acid (DHA C22:6) have different properties to those derived from the preferred substrate, arachidonic acid (ARA C20:4; n-6). Armstrong et al (2006) demonstrated that PGE2 increased progesterone when porcine granulosa cells were cultured in vitro with hCG. We hypothesized that PGE3 which is derived from EPA will produce a lower steroidogenic response as progesterone from isolated granulosa cells collected from pre-ovulatory sow ovaries. Ovaries were collected from slaughtered sows and follicles between 3–8 mm were aspirated and through a series of wash steps in HTCM (Hepes TCM 199). Mid-sized granulosa cells were recovered in a solution of BTCM (bicarbonate TCM) containing IGF-1. 0.5 × 106 cells/mL were cultured in 250 µL of Control (BTCM only), PGE2 or PGE3 (320 ng/mL in BTCM, Cayman Chemical Co.) treatments with IGF1 at 25 ng/mL. Cultures were incubated for 22 h at 38oC. Cultures were centrifuged and the supernatant was analysed in duplicate for progesterone. Data was analysed by Univariate GLM ANOVA. There was no significant difference between PGE2 and PGE3 treatments, however the main effect of PGE significantly increased progesterone production relative to the control (P = 0.017). Granulosa cells cultured with omega 3 derived PGE3 did not produce significantly lower progesterone levels than those with PGE2. We conclude that both PGE2 and PGE3 promote a steroidogenic response in cultured porcine granulosa cells. (1) Armstrong DT, Formosa, ER, Amato F, Schultz SJ. 2006. Prostaglandin E2 up-regulates luteinizing hormone receptor (LHR) expression and enhances steroidogenic responses of follicle cells.(2) Caughey GE, James MJ, Cleland LG. 2005. Prostaglandins and leukotrienes. pp. 42–49. In ‘Encyclopaedia of Human Nutrition. Vol. 4’. (Eds B Caballero, L Allen, A Prentice).

Effect of estradiol during culture of bovine oocyte–granulosa cell complexes on the mitochondrial DNA copies of oocytes and telomere length of granulosa cells

Zygote ◽  
2012 ◽  
Vol 22 (4) ◽  
pp. 431-439 ◽  
Author(s):  
M. Endo ◽  
K. Kimura ◽  
T. Kuwayama ◽  
Y. Monji ◽  
H. Iwata
Keyword(s):  
Mitochondrial Dna ◽  
Telomere Length ◽  
Granulosa Cells ◽  
Culture Medium ◽  
Mt Dna ◽  
Cell Complexes ◽  
Antral Follicles ◽  
Mitochondrial Dna Copy Number ◽  
Significant Difference

SummaryDuring the development of oocytes from early antral follicles (EAFs) to antral follicles (AFs), the mitochondrial DNA copy number (Mt DNA number) increases, and granulosa cells markedly proliferate. This study examined the effect of supplementation of culture medium with estradiol-17β (E2) on the in vitro growth of oocytes, and increases in the Mt DNA number, and telomere length during the in vitro culture of oocytes derived from EAFs (0.4–0.7 mm in diameter). The E2 supplementation improved antrum formation and the ratio of oocytes reaching the metaphase II (MII) stage, and there was a significant difference in these values between addition E2 concentrations of 10 μg/ml and 0.1 μg/ml. When the oocytes were cultured in the medium containing 10 μg/ml E2, the Mt DNA number determined by real-time polymerase chain reaction (PCR) significantly increased, and the ratio of the Mt DNA number at the end of culture to the Mt DNA number at the beginning of the culture was greatly different among cows, and could be predicted by the degree of the difference between the Mt DNA number of oocytes derived from EAFs and that of oocytes derived from AFs (3–6 mm in diameter). When oocytes were cultured for 16 days in a medium containing 10 μg/ml E2 or 0.1 μg/ml E2, the Mt DNA number of oocytes grown in vitro did not differ, but the telomere length of the granulosa cells was significantly greater in the 10 μg/ml E2 group than in the 0.1 μg/ml group. In conclusion, E2 supplementation in culture medium improved the growth of oocytes derived from EAFs, and a high E2 concentration increased the telomere length of the granulosa cells.

245 CONNEXIN 43 mRNA EXPRESSION AT DIFFERENT TIME POINT IN IN VITRO MATURATION OF BUFFALO OOCYTES

2009 ◽  
Vol 21 (1) ◽  
pp. 220
Author(s):  
S. C. Gupta ◽  
A. Pandey ◽  
N. Gupta
Keyword(s):  
Gap Junctions ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Cumulus Cells ◽  
Embryo Production ◽  
Cx43 Expression ◽  
Additional Advantage ◽  
Significant Difference ◽  
Cx43 Mrna

In advanced technologies of ART, the basic requirement is the production of in vitro-matured oocytes, and embryo production efficiency depends on healthy, matured oocytes. Oocyte growth and development depends on the ability of oocytes and their surrounding cumulus granulosa cells (Eppig et al. 1979 J. Exp. Zool. 208, 111–120). Cumulus cells provide carbohydrate precursors, amino acids, and nucleotides to the oocytes (Brower and Schultz 1982 Dev. Biol. 90, 144–153). Oocytes and cumulus cell gap junctions are required for the coordination of cytoplasmic and nuclear maturation (Carabatsos et al. 2002 Dev. Biol. 226, 167–179). In bovine COC, functional gap junctions are required for the progression of oocyte maturation. Gap junctions allow for metabolic coupling between adjacent granulosa cells. Disruption in the integrity of the gap junction inhibits oocyte maturation (Anderson and Albertini 1976 J. Cell Biol. 71, 680–686). The aim of this study was to analyze the trend of Cx43 mRNA transcript in in vitro-matured oocytes at different times of maturation in the Indian water buffalo to estimate the correlation with expression level. Oocytes collected from slaughterhouse ovaries were matured in TCM-199 medium supplemented with 2.5 mm pyruvate, gentamycin sulfate (10 mg mL–1), β-estradiol (1000 ng mL–1), FSH (500 ng mL–1), LH (500 ng mL–1), and 10% FBS at 38.5°C in 5% CO2 in air. Cumulus–oocyte complexes were used after 0, 6, 12, 18, and 24 h of maturation for the cDNA preparation with cells of a cDNA II Kit. Expression of the Cx43 gene was quantified at different time intervals for maturation with real-time PCR. Statistical analysis was performed with one-way ANOVA, followed by Duncan’s multiple pair-wise comparison. Our results showed that Cx43 mRNA abundance was affected by time of maturation. The expression of Cx43 was significantly higher at 6 h than at 18 and 24 h, whereas the 12-h value was intermediate. Our results are in agreement with decreased Cx43 protein contents in the outer cumulus layers of COC at maturation time points (Calder et al. 2003 Reprod. Biol. Endocrinol. 1, 14) and the expression of Cx43 in oocyte development regulation (Granot et al. 2002 Biol. Reprod. 66, 568–573). When Cx43 expression was compared among immature oocytes, denuded oocytes, cumulus cells, and COC at 6 h, there was no significant difference. However, 6-h-matured COC showed significantly higher expression than other groups. Further, our study supported the role of cumulus cells in COC in Cx43-mediated communication (Vozzi et al. 2001 Reproduction 122, 619–628). Differential expression of Cx43 mRNA among varying COC classes indicates that this gene may be a useful marker for oocyte quality to improve in vitro production or somatic cell nuclear transfer rates. Marker genes that predict developmental competence could be used in the optimization of maturation and culture conditions. Understanding the molecular mechanism involved in in vitro oocyte maturation would be an additional advantage in analyzing this complex biological phenomenon to improve embryo production.

The Correlation between Follicular Fluid Antimullerian Hormone Levels and Fertilization and Embryo Quality in ART Cycles

2012 ◽  
Vol 3 (3) ◽  
pp. 83-86
Author(s):  
Rutvij Jay Dalal ◽  
Akansha Mishra
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Embryo Quality ◽  
Hormone Levels ◽  
Antimullerian Hormone ◽  
Significant Difference ◽  
Antimüllerian Hormone ◽  
High Level

ABSTRACT Background Determination of oocyte and embryo quality are one of the most important goals in in vitro fertilization (IVF). Antimullerian hormone (AMH) is secreted by the ovarian granulosa cells into blood flow and follicular fluid. Follicular fluid (FF) AMH level is probably a marker of activity of granulosa cells. Objective To evaluate whether high level of FF AMH level is related to success of fertilization and better embryo quality. Materials and methods Sixty-two women, whose FF sample was obtained from a single follicle in each patient, underwent IVF with GnRH-agonist long protocol. Based on oocyte fertilization, the patients were divided into fertilized group (n = 42) and nonfertilized group (n = 20). FF AMH levels were measured in both groups and the quality of embryos was determined in fertilized group. Results Median of FF AMH level in fertilized group was higher than that in nonfertilized group (5.7 vs 2.7 ng/ml) and a statistically significant difference was observed between the two groups. There was a significant difference between FF AMH level and scores of embryos (p < 0.001). The medians levels of FF AMH were 6.7 ng/ml in good quality embryos and 3.80 ng/ml in fair quality embryos. Conclusion Our results indicate that FF AMH level has positive correlation with fertilization and embryo quality; therefore, it can be considered as a marker of IVF outcome. How to cite this article Dalal RJ, Mishra A. The Correlation between Follicular Fluid Antimullerian Hormone Levels and Fertilization and Embryo Quality in ART Cycles. Int J Infertility Fetal Med 2012;3(3):83-86.

scholarly journals UJI IN VITRO AKTIVITAS ANTIBAKTERI DARI DAUN SIRIH

2013 ◽  
Vol 2 (3) ◽  
Author(s):  
Oksfriani J. Sumampouw
Keyword(s):  
Analysis Of Variance ◽  
Leaf Extract ◽  
Piper Betle ◽  
Chemical Laboratory ◽  
E Coli ◽  
North Sulawesi ◽  
Significant Difference ◽  
Advanced Science ◽  
Drug Resistant Strains

Abstract: Diarrhea remains one of the global problems, especially in developing countries such as Indonesia. Due to drug resistant strains, new antibacterial drugs are needed to be developed. The leaf extract of piper betle from a common fruit tree in Indonesia, including Minahasa in North Sulawesi Province, is used by the Minahasans for treating diarrhea. This leaf extract has inhibitive effects on populations of bacteria and parasites. The aim of this research was to prove the antibacterial effect of piper betle leaves on Escherichia coli. This research was conducted from December 2007 to June 2008 in the Fish Product Microbiology Laboratory of the Fisheries and Marine Science Faculty at Sam Ratulangi University, the Advanced Science Laboratory of Sam Ratulangi University, and the Chemical Laboratory of Mathematics and Natural Sciences Faculty of Sam Ratulangi University. This was an in vitro experimental research. The statistical test used to verify the effectiveness of piper betle extract as an antibacterial to E. coli was the Analysis of Variance (ANOVA), followed by the Least Significant Difference test (LSD). The conclusion of this research was that the extract of piper betle leaves had antibacterial activity on E.coli. Keywords: Piper betle leaf, antibacterial.  Abstrak: Diare masih menjadi sebuah masalah global, terutama pada negara negara berkembang, termasuk Indonesia, dan khususnya Minahasa. Hal ini disebabkan beberapa strain tertentu penyebab diare seperti Escherchia coli telah resisten terhadap obat-obatan. Oleh karena itu merupakan keniscayaan ditemukan senyawa baru sebagai antibakteri untuk mengobati diare. Ekstrak daun sirih yang umumnya ada di Indonesia, juga di  Minahasa, telah sering digunakan sebagai obat anti diare. Ekstrak daun ini telah ditemukan dapat menghambat populasi bakteri dan parasit. Penelitian ini bertujuan untuk melihat aktivitas antibakteri ekstrak daun sirih terhadap E. coli. Penelitian ini dilaksanakan di Laboratorium Mikrobiologi Hasil Perikanan Fakultas Perikanan dan Ilmu Kelautan Universitas Sam Ratulangi Manado, Laboratorium Advanced Science Universitas Sam Ratulangi Manado dan Laboratorium Kimia Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sam Ratulangi Manado pada bulan Desember 2007 sampai Juni 2008. Penelitian ini merupakan penelitian eksperimental secara in vitro. Uji statistika efektifitas antibakteri dari ekstrak daun sirih terhadap E. coli menggunakan Analysis of Variance (ANOVA), dan dilanjutkan dengan uji Least Significant Difference (LSD). Hasil penelitian ini memperlihatkan bahwa ekstrak daun P. betle mengan-dung senyawa yang potensial untuk menghambat pertumbuhan E. coli Kata Kunci: daun sirih, anti bakteri.

scholarly journals Possible Intracellular Regulators of Female Sexual Maturation

2015 ◽  
pp. 379-386 ◽  
Author(s):  
A. KOLESAROVA ◽  
A. V. SIROTKIN ◽  
M. MELLEN ◽  
S. ROYCHOUDHURY
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Granulosa Cells ◽  
Sexual Maturation ◽  
Cyclin B1 ◽  
Nuclear Antigen ◽  
Dependent Protein Kinase ◽  
Significant Difference ◽  
Dependent Protein

Protein kinases, transcription factors and other apoptosis- and proliferation-related proteins can regulate reproduction, but their involvement in sexual maturation remains to be elucidated. The general aim of the in vivo and in vitro experiments with porcine ovarian granulosa cells was to identify possible intracellular regulators of female sexual maturation. For this purpose, proliferation (expression of proliferating cell nuclear antigen – PCNA, mitogen-activated protein kinases – ERK 1,2 related MAPK and cyclin B1), apoptosis (expression of the apoptotic protein Bax and apoptosis regulator Bcl-2 protein), expression of some protein kinases (cAMP dependent protein kinase – PKA, cGMP-dependent protein kinase – PKG, tyrosine kinase – TK) and cAMP responsive element binding protein 1 (CREB-1) was examined in granulosa cells isolated from ovaries of immature and mature gilts. Expression of PCNA, ERK1,2 related MAPK, cyclin B1, Bcl-2, Bax, PKA, CREB-1, TK and PKG in porcine granulosa cells were detected by immunocytochemistry. Sexual maturation was associated with significant increase in the expression of Bcl-2, Bax, PKA, CREB-1 and TK and with decrease in the expression of ERK1,2 related MAPK, cyclin B1 and PKG in granulosa cells. No significant difference in PCNA expression was noted. The present data obtained from in vitro study indicate that sexual maturation in females is influenced by puberty-related changes in porcine ovarian signaling substances: increase in Bcl-2, Bax, PKA, CREB-1, TK and decrease in ERK1,2 related MAPK, cyclin B1 and PKG. It suggests that these signaling molecules could be potential regulators of porcine sexual maturation.

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