Background: Recently, as a delayed childbearing trend is emerging in modern women’s adulthood, diminished reproductive potential due to age-related changes is more prevalent. Reduction in the abundance of mitochondrial DNA (mtDNA) copies and circulating anti-Müllerian hormone (AMH) have been separately reported with aging, contributing to the decrease in successful reproduction. However, there are limited reports on the impact of age on mtDNA and AMH in the same individual and whether mtDNA copy numbers are influenced by age and AMH. Methods: In the present study, we utilized a real-time quantitative PCR (RT-qPCR) to quantify the mtDNA copy number of granulosa cells obtained from 43 women undergoing an in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) program. Results: According to our analysis, a significant correlation was observed between age and mtDNA copy number (r = −0.54, P < 0.001) and between age and AMH level (r = −0.48, P < 0.001) of the same individual. There was also a positive correlation between mtDNA copy number and AMH (r = 0.88, P < 0.001) with AMH level falling as mtDNA decreases. In our regression, age and AMH were shown to have low collinearity (VIF = 1.297) but only AMH was correlated with mtDNA quantity (P < 0.001). Conclusion: Our study suggests that both mtDNA and AMH abundance are influenced by age and that AMH levels independently affect mtDNA copy number regardless of age. Further research is required to understand the role of AMH on mitochondria bioenergetics.

Embryo spent culture medium has been intensively investigated, considering its promising feature for non-invasive bioanalytical techniques in in-vitro fertilization (IVF). Despite, isolating DNA from such samples is quite challenging due to its small volume. Carrier RNA is reported to exhibit DNA retrieval effects and commonly employed in various limited biological samples, but there are no reports regarding its benefit on embryo media. Therefore, we aim to evaluate the competence of cRNA on isolated DNA from embryo medium and analyzed its optimal volume as there are also no records respecting its ideal volume to obtain decent outcomes. Results showed that cRNA significantly increases DNA amounts in the cRNA treated group (p<0.001), but the D-4/D-5 medium yielded similar (p=0.684). Pearson test demonstrated no correlation between cRNA volume vs. total retrieved DNA (r=0.760, p=0.80), and Whole Genome Amplification (WGA) was shown to increase DNA in the treated group (p=0.022), but not in the untreated group (p=0.128). Additionally, electrophoresis successfully resulting in a thick and thin band of TH01 locus signifies the cRNA competence. In conclusion, our study suggests that cRNA addition is essential in embryo medium extraction as it increases initial DNA that crucial for downstream application. However, the optimal volume could not be determined in the current study since the initial amount of DNA in the medium is unknown. Obtained findings are expected to be a new input for subsequent research on DNA extraction.

The fate of the corpus luteum, a transient endocrine gland formed and degraded during an oestrous cycle, is decided by various physiological factors, such as luteinizing hormone (LH). As a stimulator of progesterone, LH is known to maintain corpus luteum functional and structural integrity by inhibiting apoptosis, a programmed cell death. Therefore, we aim to investigate its action during the mid‐luteal phase hypothesized that LH suppresses the death mechanism of bovine luteal steroidogenic cells (LSC) by analysing the expression of proteins involved. Cultured bovine LSC obtained from corpus luteum were treated for 24 hr with recombinant TNF and IFNG in the presence or absence of LH. The result showed that LH proved to have a protective effect by increased cell viability (p < .05) and prevented DNA fragmentation (p < .05), as demonstrated by the WST‐1 colorimetric assay and TUNEL assay. Expression analysis of mRNA and protein levels showed that LH altered the expression of BCL2 (p < .05), CASP3 (p < .05), FAS (p < .05), and BAX (p < .05) to support cell survival. In conclusion, our study suggests that LH prolongs the corpus luteum life span through the anti‐apoptotic mechanism by increasing cell viability and suppressing apoptosis‐related genes and protein expression.

This study is aimed at establishing the effects of varying sperm quality on IVF-ICSI/IMSI outcomes in unexpected poor ovarian responder subjects of POSEIDON groups 1 and 2. In the present study, 1,263 couples with female partners who fulfilled the POSEIDON group 1 and 2 criteria were recruited. All couples underwent ICSI or IMSI at Morula IVF Jakarta Clinic, Indonesia. Patients were subsequently classified into six groups, according to semen conditions of the respective male partners: (1) normozoospermic, (2) teratozoospermia, (3) oligoasthenoteratozoospermia (OAT), (4) asthenozoospermia, (5) severe OAT, and (6) cryptozoospermia. Laboratory and clinical outcomes of the IVF-ICSI/IMSI program were then evaluated. Early and late embryonic development parameters including the number of fertilization, cleavage and blastocyst stages, and blastocyst quality differed significantly among the different sperm quality groups ( p < 0.05 ). No difference was observed in the number of embryo transfers and clinical pregnancy among the studied groups ( p > 0.05 ). Our study has demonstrated the effect of sperm quality on embryo development at the early and later stages; however, the clinical pregnancy was not impaired in the unexpected poor responders of POSEIDON groups 1 and 2.

Abstract. Bowolaksono A, Mayasari A, Sundari AM, Simamora ATJ, Suryaningsih R, Suryawan A, Simamora ATJ, Abinawanto, Sjahfirdi L, Hasna A, Supriatna J, Kyes RC. 2022. Urine biochemical profile analysis and transabdominal ultrasonography for pregnancy detection in the endangered lowland anoa (Bubalus depressicornis). Biodiversitas 23: 6056-6061. The population of lowland anoas (Bubalus depressicornis) has decreased significantly due to illegal hunting and habitat loss. As such, the urgency in developing successful ex-situ conservation programs cannot be overstated. Ex-situ breeding programs of anoas, however, are hindered by difficulties in early pregnancy detection. The failure to detect pregnancy at an early stage often results in miscarriage due to repeated breeding attempts of unconfirmed pregnant females. Moreover, frequent false heat during 5 to 6 months of pregnancy corroborated the repeated breeding of pregnant females. Therefore, early detection techniques are needed to improve breeding success. The present study evaluated the use of urine biochemical profiles as a minimally invasive approach for early pregnancy detection and transabdominal ultrasonography as a confirmation tool of false heat in four lowland anoas. Statistical analysis showed that glucose and protein level were significantly elevated during pregnancy since the 0 days of post-mating (p<0.05) and consistently higher compared to the non-pregnant period. Meanwhile, transabdominal ultrasonography successfully identified organ structures and development by the 155th, 176th, 180th, and 191st days of pregnancy. The present study suggests that urine biochemical data provide a promising, minimally invasive approach to early pregnancy detection in anoas. Incorporating urine data and ultrasonography can significantly help to maintain pregnancy by validating false heat.