To determine whether hypoxia has an effect on luteinization, we examined the influence of hypoxia on a model of bovine luteinizing and non-luteinizing granulosa cell culture. The granulosa cells were obtained from small antral follicles (≤ 6 mm in diameter). To induce luteinization, the cells were treated for 24 h with insulin (2 µg/ml), forskolin (10 µM) or insulin in combination with forskolin at 20% O2. After 24 h, progesterone (P4) production was higher in the treated cells, which we defined as luteinizing granulosa cells, than in non-treated cells, which we defined as non-luteinizing granulosa cells. P4 production by non-luteinizing granulosa cells was not affected by hypoxia (24 h at 10% and 5% O2), while P4 production by granulosa cells treated with insulin in combination with forskolin was significantly increased under hypoxia (24 h at 10% and 5% O2). Because hypoxia affected P4 production by the luteinizing granulosa cells but not by the non-luteinizing granulosa cells, hypoxia seems to promote P4 production during, rather than before, luteinization. In the cells treated with insulin in combination with forskolin, mRNA and protein expression of steroidogenic acute regulatory protein (StAR) and protein expression of 3β-hydroxysteroid dehydrogenase (3β-HSD) increased under 10% O2, while mRNA and protein expressions of key protein and enzymes in P4 biosynthesis did not increase under 5% O2. The overall results suggest that hypoxia plays a role in progressing and completing the luteinization by enhancing P4 production through StAR as well as 3β-HSD expressions in the early time of establishing the corpus luteum.

Hypoxia has been suggested to enhance progesterone (P4) synthesis in luteinizing granulosa cells (GCs), but the mechanism is unclear. The present study was designed to test the hypothesis that the hypoxia-induced increase in P4 synthesis during luteinization in bovine GCs is mediated by hypoxia-inducible factor 1 (HIF-1). GCs obtained from small antral follicles were cultured with 2 µg/ml insulin in combination with 10 µM forskolin for 24 h as a model of luteinizing GCs. To examine the influence of HIF-1 on P4 synthesis, we determined the effect of changes in protein expression of the α-subunit of HIF-1 (HIF1A) on P4 production and on the expression levels of StAR, P450scc, and 3β-HSD. CoCl2 (100 µM), a hypoxia-mimicking chemical, increased HIF-1α protein expression in luteinizing GCs. After the upregulation of HIF-1α, we observed an increase in P4 production and in the gene and protein expression levels of StAR in CoCl2-treated luteinizing GCs. In contrast, CoCl2 did not affect the expression of either P450scc or 3β-HSD. Echinomycin, a small-molecule inhibitor of HIF-1′s DNA-binding activity, attenuated the effects of CoCl2 and of low oxygen tension (10% O2) on P4 production and StAR expression in luteinizing GCs. Overall, these findings suggest that HIF-1 is one of the factors that upregulate P4 in GCs during luteinization.

Abstract. Fadhillah, Elfita, Muharni, Yohandini H, Widjajanti. 2019. Chemical compound isolated from antioxidant active extract of endophytic fungus Cladosporium tenuissimum in Swietenia mahagoni leaf stalks. Biodiversitas 20: 2645-2650. Swietenia mahagoni L. Jacq is a medicinal plant that widely used as an antimicrobial, anti-inflammatory, antimutagenic, anticancer, antitumor, and antidiabetic. Several bioactive compounds have been reported to be associated with this plant that supports its treatment activities. Flavonoid content of the S. mahagoni has antioxidant activity. Therefore the plant is widely used to treat degenerative diseases caused by free radical activity in the body. Scientists have proven that the medicinal plants have a symbiotic relationship with the number of microorganisms that can produce bioactive compounds. One of the symbiotic microorganisms is endophytic fungi believed to be associated with every plant on earth. In this paper, we report a secondary metabolite compound from endophytic fungi isolated from S. mahagoni. The endophytic fungi (DMG1-DMG4) were isolated from leaf stalks of S. mahagoni. Antioxidant activity of endophytic fungi extracts was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. The chemical compound from the active extract was isolated by the chromatographic method, and the structure was elucidated by spectroscopic methods including UV, IR, NMR 1D (1H-NMR and 13C-NMR), NMR 2D (HMQC and HMBC). Antioxidant activity test showed the endophytic fungus (DMG3) has the highest activity. The chemical compound from DMG3 was identified as 5-hydroxy-2-oxo-2H-piran-4-yl) methyl acetate). The phylogenetic method used to molecular analysis of DMG3 confirmed the fungus as Cladosporium tenuissimum.

E-modules are one of the innovations in the development of teaching materials that can be integrated with virtual laboratories. The learning model that can be applied in the development of e-modules is a guided inquiry learning model . This research aims to produce a salt hydrolisis e-module oriented on guided inquiry learning model integrated with virtual laboratories and reveal the validity and practicality of e-modules. The type of this research is research and development (R&D). the method used in this research is developmental model by using 4-D or 4-P design. They are define, design, develop and disseminate. This research is limited to the stage of development, the validity and practicality test. The data collection instrument is a questionnaire of validity and practicality. The e-module was validated by 6 experts, they are 3 chemistry lectures of FMIPA UNP and 3 chemistry teachers while practicality test was carried out by 2 chemistry teachers and 15 XII grade students of MIPA SMAS Adabiah 2 Padang. data from validity and practicality test results were analyzed using the cohen kappa formula. Based on the results of the study, it was found that average kappa moment of validity test was 0,83 with a very high validity category, the average kappa moment of teacher practicality was 0,82 with a very high practicality category and the average kappa moment students practicality was 0,82 with a very high practicality category. The results of the analysis of students answer on e-module, it was found average of key question was 90%, average of worksheet was 94,8% and average of evaluation test was 90%. Thus, it was concluded that the salt hydrolysis e-module based on guided inquiry learning virtual laboratories integrated was produced was valid and practical.

This article aims to describe the teacher's role and the implications of learning Aqidah Akhlak in the formation of the religious character of students at MTs Thoriqul Ulum Trawas. Qualitative with a case study approach has been used in this study. Data collection uses interview, observation, and documentation methods, while validity techniques use credibility, source triangulation, technical triangulation, and dependability. The results of the research are presented as follows: 1. Akhlak Aqidah teachers become motivators, mentors, and educators for students and are also less than optimal in their roles as facilitators and demonstrators. 2. The implications of Aqidah Akhlak teachers in forming the religious character of students at MTs Thoriqul Ulum, namely making students have the nature of discipline, responsibility, and honesty.