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Background: Oocyte diameter is crucial for initiating meiosis and supporting the maturation capability, a prerequisite for successful fertilisation and high-quality embryo development. While numerous studies have explored the correlation between oocyte diameter and embryo development capacity in animal models, limited research appears concerning human oocytes. Aim: This study aims to ascertain whether oocyte diameter can serve as a reliable predictor of oocyte and embryo quality, achieved through its correlation with protein kinase B (Akt) protein levels on cumulus–granulosa cells and blastocyst morphokinetics. Setting and Design: Prospective cohort study included cumulus–granulosa cells collected from 228 mature oocytes of 32 infertile women undergoing in vitro fertilisation treatment. The diameter of the oocytes was measured using the Image J software, and the samples were categorised into three groups based on their diameter: 118.97 µm. Quantification of pAkt and Akt protein total of cumulus–granulosa cells exploited the ELISA method. Embryo morphokinetics and anomaly development events were observed at the blastocyst stage to evaluate embryo quality relative to oocyte diameter. Statistical Analysis Used: The Kruskal–Wallis test and Chi-square test were used. P 0.05). Likewise, there were no significant differences in morphokinetics and anomaly development except for the S2 parameter and reverse cleavage. Conclusion: Reflecting on the molecular profile including pAkt, total Akt protein, morphokinetics and anomaly development, it was observed that oocyte diameter does not correlate with its competency and the quality of the resulting embryo. In addition, oocytes with a diameter below 105.95 µm may have an equal potential to develop into top-quality embryos.

Abstract Purpose This study aimed to evaluate the effects of immature oocyte proportion in a cohort on both IVF laboratory and clinical outcomes. Materials and methods This retrospective cohort study took place at Morula IVF Jakarta Clinic from January 2016 to July 2020. A total of 1.826 couples undergoing IVF-ICSI/IMSI were included and classified into four groups according to the proportion of immature oocytes retrieved during OPU as follows: (1) immature ≤ 15% (n = 1.064), (2) immature 16–25% (n = 369), (3) immature 26–50% (n = 331), and (4) immature > 50% (n = 62). Primary outcomes were clinical pregnancy and miscarriage. Embryology laboratory results were assessed as the secondary outcomes. Statistical analyses were carried out utilizing Kruskal–Wallis or chi-square tests. p-value

Background and Aim: Semen storage is an important reproductive method used in artificial livestock breeding. However, oxidative stress during storage reduces the quality of sperm. Melatonin supplementation in semen storage medium has not been well studied, but it has been shown to protect cells from oxidative stress. Therefore, this study aimed to determine the effect of melatonin supplementation on sperm quality parameters and antioxidant gene expression levels in semen extenders during cold storage. Materials and Methods: Semen extenders with melatonin concentrations of 0 (control), 0.1, 0.2, and 0.3 mM were added as treatment. The treated semen was then stored at 5°C for 72 h using a cold storage method, and quality parameters, including percentage of progressive motility, membrane integrity, intact acrosome, and DNA integrity, were measured every 24 h. In addition, messenger ribonucleic acid abundance levels of glutathione peroxidase (GPx) and superoxide dismutase (SOD) genes were sampled after 0 and 72 h of cold storage. Results: All observed sperm quality parameters decreased with increasing cold storage time; however, 0.2 mM melatonin demonstrated superior protection of sperm quality during cold storage. Gene expression analysis showed that GPx levels decreased significantly (p < 0.05) after 72 h in semen without melatonin but not in the melatonin-treated groups. A similar trend was also observed in SOD, indicating that exogenous antioxidants effectively protected the sperms. Conclusion: Melatonin supplementation at 0.2 mM in semen extenders during cold storage maintains sperm quality parameters for up to 72 h because melatonin protects sperm from oxidative stress. These findings can be used to improve the semen storage protocol by combining semen extender and antioxidant.

This study aims to evaluate the certainty of anti-Müllerian hormone (AMH) levels in dried blood spots (DBS) compared to serum AMH. A total of 109 infertile women who planned to undergo their controlled ovarian stimulation cycle were recruited. Serum and DBS samples were collected from each patient for the quantification of AMH levels. Serum AMH was measured with an automated clinical chemistry assay (Roche) while DBS AMH was measured using the Ansh Labs DBS AMH ELISA reagent kit. Mean AMH level in DBS was [Formula: see text][Formula: see text]ng/mL, which ranged from 0.01 to 17.3[Formula: see text]ng/mL, while the mean AMH in serum samples was [Formula: see text][Formula: see text]ng/mL (0.01–10.8[Formula: see text]ng/mL). Comparable accuracy for low AMH quantification was observed between the two methods over basal antral follicle counts (AFC) as a clinical comparator (AUC 86%, cutoff [Formula: see text][Formula: see text]ng/mL, [Formula: see text], [Formula: see text] vs. 85%, cutoff [Formula: see text][Formula: see text]ng/mL, [Formula: see text], [Formula: see text], respectively for DBS and serum sample). Likewise, DBS AMH achieved high concordance for low AMH discrimination in comparison to serum AMH method (AUC 99% vs. 93%, respectively). A strong linear Spearman regression ([Formula: see text], [Formula: see text]) and sufficient agreement (94.5%, mean difference of −1.6, 2.7 to −5.9) was demonstrated among studied groups with mathematical Passing and Bablok regression equation of [Formula: see text]. The confidence interval of intercept (−0.12, 95% CI [−0.29 to −0.01]) and slope (1.80, 95% CI [1.66–1.94]) indicated a constant and proportional difference between the two methods but it was not significant ([Formula: see text]). AMH measurement in DBS could be useful as an alternative approach to screening ovarian reserve through a more convenient remote sample collection.

Background: Epididymal sperm preservation is a simple conservation approach that can help prevent the loss of high genetic quality of farm animals. The chance of loss increases, especially during disease outbreaks or other interruptions to normal reproduction function. Aim: This study looked into the ability of preserved ram epididymal sperm to fertilize oocytes. Due to motility becoming an issue following sperm storage for fertilization, the sperm micro injection of known as intracytoplasmic sperm injection (ICSI) approach was employed. Methods: The study was divided into two parts. First, involved preservation of epididymal sperm at 5°C for 12 days. During preservation, sperm quality parameters namely motility, viability, intact membrane, acrosome, and DNA are evaluated every three days. For fertility test in the second experiment, matured oocytes were injected with immotile sperm discovered in the last days of preservation. The presence of pronucleus development following in vitro culture is used as an indicator of sperm ability to activate and fertilize oocytes. Results: All sperm quality parameters significantly (p [Open Vet J 2024; 14(2.000): 707-715]

Abstract Performance-Based Design (PBD) method is widely used to design or evaluate super high-rise building against earthquake loads. The building is expected to present a certain level of performance set on FEMA 303 in response to ground motions, and should meet the target performance at Service Level Earthquake (SLE) and at Risk-Targeted Maximum Considered Earthquake (MCER). The performance level would be determined by using non-linear time history analysis and it requires non-linier parameter based on reinforcement of the structural elements. The common method proposed by Tall Building Initiative (TBI) requires the structural members are designed using response spectra at Service Level Earthquake (SLE). The problem is the ground motion and response spectra at Service Level Earthquake (SLE) are not always immediately available. In this paper, the modified seismic response coefficient (CS-M) is introduced in designing the structural members, as an initial step of Performance-Based Design (PBD), using the common response spectra of Risk-Targeted Maximum Considered Earthquake (MCER) instead of Service Level Earthquake (SLE). The performance of buildings is evaluated at Service Level Earthquake (SLE) and Risk-Targeted Maximum Considered Earthquake (MCER) to validate that design with modified seismic response coefficient (CS-M) is still in accordance with method by Tall Building Initiative (TBI). Keywords: Modified Seismic Response Coefficient (CS-M), Performance-Based Design (PBD), Risk-Targeted Maximum Considered Earthquake (MCER), Service-Level Earthquake (SLE), Tall Building Initiative (TBI). Abstrak Metode Desain Berbasis Kinerja (PBD) banyak digunakan untuk merancang atau mengevaluasi bangunan super tinggi terhadap beban gempa bumi. Bangunan diharapkan dapat mencapai tingkat kinerja tertentu yang ditetapkan dalam FEMA 303 sebagai respons terhadap gerakan tanah, dan harus memenuhi kinerja target pada Service Level Earthquake (SLE) dan pada Risk-Targeted Maximum Considered Earthquake (MCER). Tingkat kinerja akan ditentukan dengan menggunakan analisis riwayat waktu non-linier dan membutuhkan parameter non-linier berdasarkan penulangan elemen struktural. Metode umum yang diusulkan oleh Tall Building Initiative (TBI) mengharuskan komponen struktural dirancang menggunakan respons spektra pada Service Level Earthquake (SLE). Masalahnya adalah gerakan tanah dan respons spektra pada Service Level Earthquake (SLE) tidak selalu tersedia secara langsung. Dalam paper ini, diperkenalkan koefisien respons seismik yang dimodifikasi (CS-M) dalam merancang komponen struktural, sebagai langkah awal Desain Berbasis Kinerja (PBD), dengan menggunakan respons spektra umum dari Risk-Targeted Maximum Considered Earthquake (MCER) sebagai ganti dari Service Level Earthquake (SLE). Kinerja bangunan dievaluasi pada kondisi Service Level Earthquake (SLE) dan Risk-Targeted Maximum Considered Earthquake (MCER) untuk memvalidasi bahwa desain dengan koefisien respons seismik yang dimodifikasi (CS-M) masih sesuai dengan metode yang diajukan oleh Tall Building Initiative (TBI). Kata kunci: Koefisien Respons Seismik yang Dimodifikasi (CS-M), Desain Berbasis Kinerja (PBD), Risk-Targeted Maximum Considered Earthquake (MCER), Service-Level Earthquake (SLE), Tall Building Initiative (TBI).

Background and Aims: Embryo aneuploidy is caused by many factors, including patient age, sperm quality and sperm selection method. There are several methods for selecting sperm, ICSI (intracytoplasmic sperm injection) and IMSI (intracytoplasmic morphologically selected sperm injection). Previous study related to the effect of sperm selection methods on aneuploidy embryos rate is still very rare. This study aims to determine the effect of selecting sperm methods on the aneuploidy embryos rate, especially in the good prognosis patient group. Method: A total of 67 patients with a good prognosis (AFCs[Formula: see text]6; AMH[Formula: see text]1,2) were grouped based on the method of selecting sperm. Furthermore, the fertilized oocytes that become blastocyst embryos were biopsied in the trophectoderm of the cell and compared between the two treatments. Results: The results of the study showed that the sperm selection method did not significantly affect the aneuploidy embryos rate (P>0.05). The percentage of aneuploidy embryos in the ICSI method was 28.1%, while in the IMSI method it was 24.3%. Conclusion: The conclusion of this study is that the sperm selection method does not affect the aneuploidy embryos rate in patients with a good prognosis.

Abstract Background: Spinal muscular atrophy (SMA) is characterized by the homozygous deletion of the survival motor neuron-1 gene. Pre-implantation genetic testing for monogenic diseases through in-vitrofertilization program was developed to provide a reliable genetic diagnostic method for SMA. Case presentation: The couple who was confirmed as carriers of SMA visited the Morula IVF Clinic, Jakarta, Indenesia seeking for an in-vitro fertilization expert opinion in relation to the pre-implantation genetic testing for SMA. Utilizing polymerase chain reaction-restriction fragment length polymorphism, we have successfully screened for unaffected embryos that were characterized by a normal presence of the survival motor neuron-1 exon 7-8 and survival motor neuron-2 exon 7-8. The frozen embryo was subsequently transferred and a healthy unaffected female baby was born with undetected deletion of the survival motor neuron-1 gene. Conclusion: This successful embryo pre-implantation screening case could potentially accommodate the demands of genetically at-risk couples who are apprehensive about conceiving a child who might inherit monogenic disorders such as SMA.

BACKGROUND: Preimplantation genetic testing for aneuploidy has been proven to be effective in determining the embryo's chromosomal or ploidy status. The test requires a biopsy of embryonic cells on day 3, 5, or 6 from which complete information on the chromosomes would be obtained. The main drawbacks of preimplantation genetic testing for aneuploidy include its relatively invasive approach and the lack of research studies on the long-term effects of preimplantation genetic testing for aneuploidy. OBJECTIVE: Computer-assisted predictive modeling through machine learning and deep learning algorithms has been proposed to minimize the use of invasive preimplantation genetic testing for aneuploidy. The capability to predict morphologic characteristics of embryo ploidy status creates a meaningful support system for decision-making before further treatment. STUDY DESIGN: Image processing is a component in developing a predictive model specialized in image classification through which a model is able to differentiate images based on unique features. Image processing is obtained through image augmentation to capture segmented embryos and perform feature extraction. Furthermore, multiple machine learning and deep learning algorithms were used to create prediction-based modeling, and all of the prediction models undergo similar model performance assessments to determine the best model prediction algorithm. RESULTS: An efficient artificial intelligence model that can predict embryo ploidy status was developed using image processing through a histogram of oriented gradient and then followed by principal component analysis. The gradient boosting algorithm showed an advantage against other algorithms and yielded an accuracy of 0.74, an aneuploid precision of 0.83, and an aneuploid predictive value (recall) of 0.84. CONCLUSION: This research study proved that machine-assisted technology perceives the embryo differently than human observation and determined that further research on in vitro fertilization is needed. The study finding serves as a basis for developing a better computer-assisted prediction model.

Cryopreservation is a technique for storing cells and tissues at very low temperatures for the possible usage of the stored cells and tissues throughout the year. Sperm cell cryopreservation in some species causes a decrease in sperm quality and DNA damage. Inappropriate cryopreservation protocols can cause changes in sperm physiology. Mitochondria are organelles that play a role in producing energy for sperm motility. Mitochondria have DNA molecules with small sizes compared to structures from nuclear DNA. This study analyzed the effect of cryopreservation on sperm motility and the Cytochrome Oxidase1 (COI) gene. The CO1 gene in mitochondrial DNA plays a role in energy production for spermatozoa motility. The cryopreservation was performed using skim milk and 10% methanol cryoprotectant, and the temperature in the equilibration process was 4-5°C for 10 minutes. Cryopreservation took place for 14 days in the freezer at -80°C. In addition, the thawing process was performed for 1-2 minutes at 40°C. This study found that the number of lesions per 10 kb in the CO1 gene in post-equilibration spermatozoa was (9.24±3.74), and post-thawing spermatozoa (was 10.26±7.54). Spermatozoa motility was obtained in fresh spermatozoa, i.e., 87±1.5%, post-equilibration spermatozoa 79±4.5%, and post-thawing spermatozoa 30±3.2%. This study concluded that cryopreservation of spermatozoa causes CO1 gene lesions and that in cryopreserved spermatozoa, there is a decrease in spermatozoa motility compared to fresh spermatozoa.

Background and Aims: Spermatozoa morphology is one of the factors that can influence the success of IVF. IMSI is a technique that provides various selection criteria of sperm head’s morphology and the presence of vacuoles with a total magnification of over 6,000. The purpose of this study was to compare the embryo development of good prognosis patients based on sperm morphology grading using IMSI technique. Method: This study was a retrospective analysis performed at Morula IVF Jakarta from 2019 to 2022 based on good prognosis patients (age [Formula: see text] 38 years, AFC [Formula: see text] 7, AMH [Formula: see text] 1.1). There were two groups in this study, (A) sperm grade I and II, (B) sperm grade III and IV, based on Vanderzwalden et al. (2008). The number of patients in group (A) was 1035 patients, and group (B) was 345 patients. The parameters observed in this study were fertilization rate (FR), top embryo quality rate on day 3 (D3) and day 5 (D5), blastocyst rate (BR). Results: The result study showed that (FR) were significantly increased (P=0.000) in group (A) compared to group (B), which was 77.28% and 72.90%. In addition, group (A) had significantly (D3), which was 51.37% (P=0.012) compared to group (B) 46.92%. On the other hand (D5) and (BR) were significantly increased (P=0.000) in group (A), by about 42.58% and 48.01%, contrasted to group (B), which was only 36.11% and 39.21%. Conclusion: Grade I and II sperm of IMSI technique significantly increased IVF outcome in good prognosis patient.

Background and Aims: Frozen Embryo Transfer is the final step in undergoing IVF treatment. One of the important parts in the success of the FET cycle is the quality of the transferred embryos. The ability of blastocyst post-thawed recovery indicated embryo potential. However, there is limited understanding of the factors that influence recovery of post-thawed blastocyst embryos. This study aims to examine whether blastocyst morphology post-thawed correlates with clinical pregnancy rate. Method: In a retrospective cohort study, analysis data of Morula IVF Surabaya was conducted from January 2022 – February 2023, thawed blastocyst with a post-thaw cell survival rate of 50%–100% were observed for morphological characteristics prior to transfer. Then, recovery of blastocoel expansion was graded as A, fully expanded; B, re-expanded; and C, not expanded. Results: The primary outcome was the clinical pregnancy rate per cycle. In the 134 cycles reviewed, the clinical pregnancy rate (CPR) reached 5.5% of the not expanded; 29.4% of the re-expanded; and 56.92% of the full expanded blastocyst group (p

Background and Aims: Calcium stored and released by the Smooth Endoplasmic Reticulum - SER which has an important role in oocyte maturation, fertilization, and embryo development. The presence of SER aggregate result in the disturbance of calcium stores and its occillation which in turn affect fertilization, embryo develompent, and increasing risk of abnormal cromoseme outcome. Alpha-ESHRE Consensus (2011), strongly recomended to not perform insemination on any oocytes with SER. In the other hand, some previous studies show that healthy babies had been born from oocytes presenting this dymorphism. The aim of this study was to evaluate embryo quality and pregnancy outcome patients derived from oocytes with SER. Method: This study was a crosssectional analysis from 2017-2019 of 223 MII oocytes with SER from 82 women underwent Intra cytoplasmic sperm injection - ICSI. Prior to ICSI, each oocyte was evaluated for the presence of SER Aggregates. The embryological outcome measures were the fertilization rate, embryo quality day 3 and day 5, and pregnancy rates. Results: This study result shown that from 223 oocytes with SER, 89.5% oocytes can be fertilized. Day 3 embryo 66.6% ± 0.000 qualified as good embryo, but only 29.1% ± 0.000 on day 5 embryo. The Pregnancy rate was 30.0%, with 2 miscarriages. It is important to give a well information to the patients on the risk of birth malformations as well as whether a decreased chance of pregnancy exists. Conclusion: Oocyte with SER still has the ability to develop and has the potential to support pregnancy.

Embryologist is an essential part of infertility care defined as a highly skilled and well-trained person who can work in teamwork to execute continuous tasks with near-zero error and pay attention to detail with excellent clinical judgment. At the very beginning, embryologist’s routine daily tasks were quite simple including processing the sperm and eggs for natural insemination. Following that, advanced infertility treatment emerged quickly including intracytoplasmic sperm injection (ICSI), vitrification-warming, embryo shipping, intracytoplasmic morphologically sperm injection (IMSI), as well as trophectoderm biopsy as a source of chromosomal copy number testing or monogenic disorders analysis. Collectively, the implementation of such improved treatments renders complexity in the IVF laboratory that could be simplified in the future. Apparently, the implementation of advanced technology in the field of IVF nowadays has two main purposes. The first is to improve the IVF success rate including finding precise markers for embryo selection such as genomics, proteomics, and metabolomics fingerprint and images-based artificial intelligence algorithm for embryo selection. The second is to standardize or simplify routine daily tasks in the embryology laboratory by implementing automated software such as for laboratory quality control maintenance, a microfluidic system that allows standardization of handling gametes, micro-robotic for immobilizing sperm for ICSI, fully automated semen analysis, electronic tagging of laboratory devices to improve traceability. All such automated systems will serve as advanced clinical decision tools to boost IVF success rate and infertility care quality. An elevated automated system in the future in an embryology laboratory would impact the clinical embryologist in terms of reduced quantity and conversely, increased qualities of highly scientific embryologists with proper education. Embryologist roles may change from conventional routine daily tasks to well-trained and experienced embryologists who are able to scientifically evaluate the effectiveness of certain approaches or potential clinical use of such implementation in their clinical laboratory setting.

BACKGROUND AND AIMS: Infertility in males contributes to about 40% of cases of infertility in couples, either as the main factor or along with female factors. Fragmentation of sperm DNA is known as a cause of male infertility, which impacts the failure of couples to undergo assisted reproductive programs, such as embryo development and recurrent miscarriage. However, some literature studies are still debating this. This research aims to determine the correlation between sperm DNA fragmentation and IVF laboratory performance. METHODS: Infertile couples undergoing an in vitro fertilization program at Morula IVF Jakarta in 2022 are the samples for this study. We divided them into two groups: good prognosis (maternal age [Formula: see text]38 years, AFC[Formula: see text]7; AMH[Formula: see text]1.1) and poor prognosis (maternal age > 38 years, AFC 14.9% had a significant negative correlation with IVF laboratory performance, particularly in the fifth-day embryo quality and blastocyst rate in the good prognosis group. Whereas in the group with poor prognosis, all laboratory performance parameters were not affected by high sperm DNA fragmentation values. CONCLUSIONS: Sperm DNA fragmentation rate > 14.9% correlated with fifth-day embryo quality and a lower blastocyst rate in the good prognosis group.

Objective: Hidden knowledge could be discovered within a large practical data of in vitro fertilization (IVF) practice. In this study, Machine learning-based data mining techniques were utilized to construct a reliable prediction model for clinical pregnancy in IVF. Study Design: A retrospective cohort multicenter study involving 4.570 IVF cycles. All patients underwent fresh embryo transfer at either the cleavage or blastocyst stage between January 2015 and December 2019. The experiment focused on utilizing tree-based classifiers to generate and compare the most effective prediction model that could predict a clinical pregnancy through clinical data. Additionally, each classifier is optimized via a genetic algorithm technique, along with the selection of variables. Results: Both the decision tree and random forest showed similar performance that was much better than the gradient boost. The two superior classifiers achieved a balanced accuracy of roughly 0.62. Additionally, each prediction model was shown to work optimally with different combinations of variables, with some variables being consistently included, such as female age, and some consistently excluded, which provides an insight into the relationship between the variables and each prediction model. Conclusion: Machine learning algorithm remains effective for the purpose of data mining and knowledge extraction in IVF clinical datasets through which a relatively reliable prediction system for clinical pregnancy could be constructed, provided the available data is sufficient.

Injection of female mammalian animals with gonadotropin prior to mating using superovulation dose increases estradiol 17beta (E2) synthesis and secretion and improves the growth and development of the uterus to support prenatal growth with the side effect of increasing litter size. This experiment was designed to study the effects of low doses of PMSG injection in heifers on the growth of granulose cells (GC) of pre-ovulatory follicles, E2 synthesis, and uterine glands growth and development. Nine Ongole-grade heifers were divided into three groups of doses of PMSG injection, i.e., 0 IU/kg BW, 0.5 IU/kg BW, and 1.0 IU/kg BW. The injection of pregnant mare serum gonadotropin (PMSG) was conducted at the early stage of the second follicle wave. Blood samples were collected at estrus (day 0) to measure plasma E2 concentrations. The ovary and uterus were collected for histological observation after slaughtering the animal. The results showed that non-superovulation doses of PMSG injections at the second follicle wave significantly increased the growth of GC that eventually produced a significantly higher plasma E2 concentration by 39.47% and 113.57%, in heifers receiving 0.5 IU/kg BW and 1.0 IU/kg BW (p

BACKGROUND: A clinical pregnancy prediction model was developed by implementing machine learning technology that uses a combination of static images and medical data to calculate the outcome of an in vitro fertilization cycle.OBJECTIVE: To provide a system that can accurately and sufficiently assist with decision making that is critical to in vitro fertilization cycles, primarily embryo selection.STUDY DESIGN: Historical medical data, which consist of clinical information and a complete transferred embryo image dataset, of 697 patients who underwent unique in vitro fertilization were collected. Various techniques of machine learning were used, namely decision tree, random forest, and gradient boosting; each technique used the same data configuration for performance comparison and was subsequently optimized using genetic algorithm.RESULTS: A prediction model with a peak accuracy of approximately 65% was achieved. Significant differences in the performances of the 3 selected algorithms were apparent. Nonetheless, additional metric measurements, such as receiver operating characteristic, area under the receiver operating characteristic curve score, accuracy, and loss, suggested that the gradient boosting model performed the best in predicting clinical pregnancy.CONCLUSION: This study served as a stepping stone toward the application of in vitro fertilization prediction models that use machine learning techniques. However, additional validation steps are required to boost the model's performance for its implementation in the clinical setting.

Abstract Background: The umbilical cord-derived mesenchymal stem cells conditioned medium (UC-MSCs-CM) produces secretomes with anti-apoptotic properties, and has the potential to prevent apoptosis of granulosa cells (GC) during controlled ovarian hyperstimulation. Objective: To observe the effect of UC-MSCs-CM on the interaction between pro- and anti-apoptotic proteins and the influence of growth differentiation factor 9 (GDF9) production in GC. Materials and Methods: UC-MSCs-CM was collected from umbilical cord stem cell culture on passage 4. GC from 23 women who underwent in vitro fertilization were cultured and exposed to UC-MSCs-CM for 24 hr. Then RNA of the GC was extracted and the mRNA expression of BCL-2 associated X (BAX), survivin and GDF9 were analysed using quantitative real-time PCR. The spent culture media of the GC were collected for measurement of insulin growth factor 1 using ELISA. Results: The expression of BAX was significantly different after UC-MSCs-CM exposure (4.09E-7 vs. 3.74E-7, p = 0.02). No significant changes occurred in survivin, BAX/survivin ratio, and GDF9 expression after UC-MSCs-CM exposure (p > 0.05). The IGF-1 level of the CM was significantly higher after the CM was used as a culture medium for GC (2.28 vs. 3.07 ± 1.72, p ≤ 0.001). A significant positive correlation was found between survivin and GDF9 (r = 0.966, p ≤ 0.001). Conclusion: IGF-1 produced by UC-MSCs-CM can work in paracrine fashion through the IGF receptor, which can inhibit BAX and maintain GDF9 production. Moreover, under the influence of UC-MSCs-CM, GC are also capable of producing IGF-1, which can impact GC through autocrine processes.

Our study aims to assess the utilization of oocyte cryopreservation (OC) in Indonesian women. Data from 122 women who had undergone OC was retrospectively analyzed from medical records. The baseline profile, clinical characteristics, and main outcomes comprising intentions for oocyte vitrification and outcomes following oocyte warming were examined. Out of 122 women who underwent OC, 49 patients returned and use their cryopreserved oocytes with a median duration of storage was two months. Arranged from the greatest to the least, participants had undergone the cycle of OC due to sperm factor [51 (41.80%)], increased embryo availability [6 (12.1%)], postponement of marriage [6 (12.1%)], social reasons [10 (8.19%)], and other reasons [8 (6.55%)]. Meanwhile, treatment due to advanced maternal age [6 (4.91%)], poor ovarian reserve [6 (4.91%)], cancer [5 (4.09%)], PCOS [2 (1.63%)], and endometriosis [2 (1.63%)] was reported among remaining subjects. Clinical pregnancy was reported in 12 (40.0%) patients constituting of each 6 (50.0%) subjects of day-3 and day-5 embryo transfer, respectively. Our study demonstrated that sperm factor, increased embryo availability, and postponement of marriage is the main reason for women undergoing OC in Indonesia.

Abstract: Students' understanding can be improved through the use of learning media in the form of applications that can be accessed via smartphones. This study tries to develop mathematics learning media in the form of educational games using the Construct 2 application. The type of research used is Research and Development (R&D) which refers to the Thiagarajan model which consists of define, design, development stages, and dissemination. The educational games that have been designed are assessed by media experts, material experts, and students as users of learning media. The educational game produced is in the form of a Mathventure Game with features tailored to the needs of elementary school students. To assess the feasibility of the product, individual, small group, and large group trials were conducted. The results showed that the Mathventure Game developed was valid and feasible to use. Further development can be done with more variety and take some basic competencies. Abstrak: Salah satu cara meningkatkan pemahaman siswa adalah dengan media pembelajaran dalam bentuk aplikasi yang dapat diakses melalui smartphone. Penelitian ini mencoba mengembangkan media pembelajaran matematika dalam bentuk game edukatif menggunakan aplikasi construct 2. Jenis penelitian yang digunakan adalah Research and Development (R&D) yang mengacu pada model Thiagarajan yang yang terdiri dari tahap define (pendefinisian), design (perancangan), development (pengembangan) dan dissemination (penyebaran). Game edukatif dinilai oleh ahli media, ahli materi, dan siswa sebagai pengguna media pembelajaran. Game edukatif yang dihasilkan berbentuk Game Mathventure dengan fitur yang disesuaikan dengan kebutuhan siswa tingkat Sekolah Dasar. Untuk menilai kelayakan produk, dilakukan uji coba perseorangan, uji coba kelompok kecil dan uji coba kelompok besar. Hasil penelitian menunjukkan Game Mathventure yang dikembangkan telah valid dan layak untuk digunakan. Pengembangan lebih lanjut dapat dilakukan dengan lebih variatif dan mengambil beberapa kompetensi dasar.

Human Wharton’s jelly‐mesenchymal stem cells (hWJ‐MSC) are an emerging potential source of stem cells derived from the umbilical cord. Previous studies have shown their potential as treatment for traumatic brain injury and Parkinson’s disease. However, no study has yet investigated the effect of hWJ‐MSC injections in countering spatial learning and memory impairment in aging rats. The effect of hWJ‐MSC injection on young rats is also unknown. The objective of this research was to analyze the effect of an hWJ‐MSC injection on spatial learning, memory, density of putative neural progenitor cells (pNPC), and neuronal apoptosis in the dentate gyrus (DG) of young and aging rats. Injection of hWJ‐MSC did not change spatial learning and memory in young rats until two months post‐injection. This might be due to retained pNPC density and neuronal apoptosis in the DG of young rats after injection of hWJ‐MSC. In contrast, injection of hWJ‐MSC promoted both spatial learning and memory in aging rats, a finding that might be attributable to the increased pNPC density and attenuated neuronal apoptosis in DG of aging rats during the two months post‐injection. Our study suggests that a single injection of hWJ‐MSC might be sufficient to promote improvement in long‐term learning and memory in aging rats.

BackgroundMesh-augmented surgery with polypropylene meshes (PPMs) is often used in urogynecology and pelvic reconstructive surgery. However, the various complications that arise from its integration process have resulted in a decrease in the number of mesh-augmented surgeries performed worldwide. An approach to improving mesh-tissue integration is coating PPMs with anti-inflammatory and wound-healing molecules, such as platelet-rich plasma (PRP), which is a component of biotechnologies that are capable of accelerating wound healing. Estrogen is also known to have a beneficial effect on wound remodeling; therefore, a hypoestrogenic status may have negative implications for wound healing. The mechanism of how PRP plays a role in wound remodeling, especially among individuals in a hypoestrogenic state, has not been fully described until now. ObjectiveOur aim is to investigate the impact of applying PRP to PPMs in hypoestrogenic rabbit models. MethodsOur study will be a randomized controlled trial involving hypoestrogenic rabbit models. Samples were categorized into either the PRP group or the PPM group (1:1 ratio), with a minimum sample size of 16 in each arm, via simple random sampling. All samples were put into a hypoestrogenic state via bilateral oophorectomy. After confirming a decrease in estradiol level, the meshes were implanted in the vesicovaginal space. The samples were euthanized on the 14th, 28th, or 90th day of the surgery. The mesh-tissue integration process will be analyzed based on inflammatory parameters (inflammatory infiltrate, interleukin-17, and interleukin-1B expression); angiogenesis (CD31 expression); and collagen deposition, which will be assessed by using Masson trichrome staining. ResultsOur study is in the protocol development stage. A preliminary study regarding its feasibility, including the feasibility of the preparation of hypoestrogenic rabbit models, mesh implantation in the rabbits’ vesicovaginal spaces, the PRP and amnion scaffold, started in February 2022. The results of our study are expected to be available by the end of 2022. ConclusionsOur randomized controlled trial is designed to provide high-quality evidence on the effect of applying a PRP-decellularized amnion scaffold to PPMs in the vesicovaginal spaces of hypoestrogenic rabbit models. International Registered Report Identifier (IRRID)PRR1-10.2196/37942

External pH is crucial in preserving sperm viability and ensuring fertilization during in vitro conditions. The purpose of this study was to determine the maximum pH value that can be tolerated by frozen-thawed Pasundan bull sperms and the effect on sperm quality. Around 250x106 sperms/mL of frozen-thawed Pasundan bull sperms were divided into ten equal aliquots, and each was diluted in the medium within a particular pH value. HCL or NaOH was added to the buffer media to create ten different solutions with varying pH values of 3, 4, 5, 6 as acidic, 7.2-7.4 as a control, and 8, 9, 10, 11, and 12 as alkaline. Furthermore, the samples were incubated for 5 minutes at room temperature within a particular pH medium before being immediately supplemented with a buffered medium to achieve a pH of 7.2-7.4. After 10 minutes of incubation at room temperature, all parameters were assessed. The results showed that sperm motility, viability, normal morphology, and acrosome intactness in sperms incubated in the acidic or alkaline media were significantly lower compared to control (p

Objective: To determine the correlation of different serum estradiol levels on the trigger day with the clinical and laboratory outcomes of in-vitro fertilization (IVF) cycles comprising a single fresh top-quality blastocyst transfer. Methods: This was a retrospective observational study performed in Morula IVF Clinic Jakarta. Five hundred forty-two women were recruited and grouped according to their serum estradiol levels on the trigger day of follicular maturation as follows: 0.05). Nonetheless, the study demonstrated a positive correlation of the serum estradiol levels with the overall laboratory outcomes including the number of retrieved, mature, and fertilized oocytes, the total sum of derived embryos, and top-quality embryos at cleavage and blastocyst stage (P

Background:Luteinizing hormone (LH) supplementation may have beneficial effect on the maturity and fertilizability of oocytes in poor ovarian reserve (POR) and may influence the progesterone level, thus increasing the pregnancy rate. However, previous studies on the effect of LH activity supplementation on poor responders have shown conflicting results. This study aimed to compare the clinical effectiveness of two different forms of gonadotropin (highly purified human menopausal gonadotropin (HP-HMG) vs. recombinant human follicle-stimulating hormone (r-hFSH)-only) in Indonesian population. Methods: Women diagnosed with poor ovarian response who received gonadotropin-releasing hormone (GnRH) antagonist protocol with either HP-HMG or r-hFSH-only were investigated. Women who underwent freeze all cycles, mini stimulation, and natural stimulation were excluded. Multiple logistic regression was performed to assess the effect of follicle-stimulating hormone (FSH) + human chorionic gonadotropin (HCG)-driven LH activity combination in HP-HMG to pregnancy event adjusting for progesterone level, demographic variables, and clinical characteristic variables. Results: A total of 101 subjects in the HP-HMG treatment group and 89 subjects in r-hFSH-only treatment group were involved in the study. There was no significant difference of clinical pregnancy rate between HP-HMG group and r-hFSH-only group (adjusted OR: 0.94, 95% CI: 0.39–2.25; p-value: 0.890). Conclusion: Compared to r-hFSH-only group, combination of FSH + HCG-driven LH activity in HP-HMG group had similar effectiveness in poor responders undergoing in vitro fertilization (IVF) using the antagonist protocol.

Objective: To assess the effect of short-term gavage of ethanolic extract of Imperata cylindrica L root on the ovarian activity and estrus behavior of female mice. Methods: Eighteen virgin female ddY mice, 8 to 10 weeks of age, weighing 22–25 g with regular estrus cycle, were divided into three groups. Group 1 received 0.5% carboxymethylcellulose, whereas Groups 2 and 3 received the ethanolic extract of Imperata cylindrica L root at 90 and 115 mg/kg body weight (b.w.) per day by gavage for 20 days, respectively. All of the groups were checked before 9 a.m. daily for vaginal cytology to determine the estrus phase. On day 21, the mice were sacrificed to collect serum samples to quantify the concentrations of reproductive hormones using enzyme-linked immunosorbent assay and to determine changes in the reproductive organs based on their reproductive organ weight, histomorphology, and histomorphometry of ovarium and uterus. Results: The reproductive organ weight in the treatment groups was similar compared with that in the control group. The 90 mg/kg b.w. treatment group showed an increase in corpus luteum number when compared with the control group, with few degenerated follicles and diminished oocytes. Moreover, the 115 mg/kg b.w. treatment group showed fewer primordial and primary follicles and an increase in corpus luteum number and a prolonged diestrus phase compared to the control and 90 mg/kg b.w. treatment groups. The histomorphology examination of the uterus showed that the thickness of myometrium and epithelium in the treated animals was similar to the control group. In addition, there was a significant decrease in follicle-stimulating hormone level in the 115 mg/kg b.w. treatment group (P

Objective: To compare the effectiveness of blastocyst elective single embryo transfer (eSET) and double embryo transfer (DET) in reducing low birth weight, preterm birth, and perinatal mortality in in vitro fertilization (IVF) cycles of Indonesian women. Methods: A retrospective observational study was conducted at Morula IVF Clinic, Jakarta, Indonesia. A total of 179 women who underwent either eSET or DET and had met the eligibility criteria were included. Seventy-six women underwent eSET while 103 underwent DET in their IVF cycles. Low birth-weight rate, preterm birth rate, and perinatal mortality rate of both groups were measured as the primary study outcomes. Neonatal intensive care unit (NICU) admission rate, Apgar score, multiple pregnancy, and maternal complications during pregnancy were also evaluated. Results: The risk of low birth weight [odds ratio (OR)=0.21, 95% confidential interval (CI): 0.10-0.45, P< 0.001] and preterm birth (0R=0.25, 95% CI: 0.13-0.49, P< 0.001) was significantly lower in the eSET group compared with the DET group. Furthermore, eSET efficiently reduced the incidence of NICU admission and multiple pregnancy (P=0.01 and P< 0.001, respectively). No significant difference was observed in terms of perinatal mortality rate, Apgar score, and maternal complications including gestational diabetes, preeclampsia as well as pregnancy-induced hypertension (P>0.05). However, a lower incidence of antepartum hemorrhage was noticed in the eSET group than in the DET group (P=0.03). Conclusions: Compared with DET, infants conceived through IVF cycles with eSET have a significantly lower risk of low birth weight, preterm birth, and NICU admissions. Moreover, eSET is shown to reduce multiple pregnancy rate, yet no significant differences are observed in the perinatal mortality rates, Apgar score and maternal complications (except for the incidence of antepartum hemorrhage) between both groups.

Background: Ca2+ signaling pathway is suggested to play an essential role in mediating oocyte maturation. Aims: The aim of this study was to evaluate intracellular Ca2+ of resistant immature oocytes that failed to resume meiosis following subsequent in vitro culture reach metaphase II after calcium ionophore A23187 activation. Settings and Design: This in vitro analytical experimental study was conducted at Animal Science Laboratory of Indonesian Medical Education and Research Institute (IMERI), Human Reproductive Infertility and Family Planning of IMERI, and Electrophysiology Imaging of Terpadu Laboratory, Faculty of Medicine, University of Indonesia. Methods: A total of 308 oocytes classed as resistant immature following in vitro culture were randomly allocated to control (n = 113) and treatment groups (n = 195). The oocyte activation group was exposed to A23187 solution for 15 min and then washed extensively. Maturation was evaluated by observing the first polar body extrusion 20‒24 h after A23187 exposure. Ca2+ imaging was conducted using a confocal laser scanning microscope to identify the dynamic of Ca2+ response. Statistical Analysis: SPSS 20, Chi-square, and Mann–Whitney U-test were used in this study. Results: Activation of resistant immature oocytes with A23187 significantly increased the number of oocyte maturation compared with the control group (P

Interspecies embryo transfer is a seldom-used method to increase the successful conservation of endangered species. The study aimed to determine the potential development of interspecies reconstructed embryos. The present study used two animal models, Campbell hamsters (Phodopus campbelli) and mice (Mus musculus). The isolated inner cell mass (ICM) of hamster embryos were injected into the mice embryos. The embryos were transferred to the pseudopregnant mice using non-surgery embryos transfer methods. The fetuses were collected at day 13.5 of gestation for morphometric measurement and cytochrome b (Cyt b) analysis which used to determine the species of obtained fetuses. The results showed that the viability, pregnancy rate, and embryonic implantation ability of the interpecies reconstructed embryos did not differ significantly (p>0.05) compared to non-reconstructed embryos. Morphometric measurement showed that the crown-rump (CR) and the weight of fetuses in the reconstructed group were significantly higher than non-reconstructed group (p

Bone marrow mesenchymal stem cells have been widely used, because plasticity, specific surface markers, self-renewal to transform into various lineages including cardiomyocytes. Information about the connexin (Cx) cardiac conduction systems of the pigtail macaque (Macaca nemestrina) is limited. This study aimed to evaluate cardiomyocyte differentiation from bone marrow mesenchymal stem cells of pigtail macaques and to clarify the Cx cardiac conduction system. Bone marrow aspirates were obtained from the proximal humerus of four adult male pigtail macaques, collected into heparinized tubes, then centrifuged to obtain mononuclear cells that were isolated and cultured in an incubator. After these cells reached 70–80% monolayer confluency as homogeneous fibroblast-like cells, they were subcultured. On the second subculture passage, the cells were pelleted to extract the mRNA, which was analysed by reverse transcription–polymerase chain reaction, and then cultured for a third passage. Cells were positive for CD73 and CD105 and the reference gene glyceraldehyde-3-phosphate dehydrogenase, and negative for CD34 and CD45. Osteogenic, chondrogenic, adipogenic, and cardiomyocyte differentiation was confirmed based on specific staining. The pigtail macaque bone marrow mesenchymal stem cells can be isolated and subcultured. The transcription of genes and translation of proteins of the connexin cardiac conduction systems was successfully identified.

Parthenogenesis is an artificial oocytes activation process without paternal contribution. Blastocyst, derived from parthenogenesis, is one of potential source for pluripotent stem cell propagation. Unfortunately, previous studies reported that parthenogenetic embryo did not achieve exhilarating blastocyst rate. One of the component that predicted inhibit parthenogenetic embryo development is phosphate. Therefore, we try to modify culture medium in order to overcome that problem. The aim of this research was to produce and analyze the characteristics of parthenogenetic blastocyst developed in phosphate-free medium. Mouse oocytes obtained from adult female DDY by superovulation. The activator was strontium chloride 10 mM and diploidization with cytochalasin B 5 μg/ml. Medium for activation and culture medium were modified rat 1 cell embryo medium (MR1ECM) which is phosphate free. The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups. The increase of phosphate in culture medium lead to impaired parthenogenetic embryos development. Further experiment was made to analyze the differences between fertilized and parthenogenetic embryo in this medium. The experiment showed that diploid parthenogenetic could achieve high blastocyst rate (30.9±1.3%). The quality of diploid parthenogenetic blastocyst, based on cells number, viability, and ICM ratio, was lower than fertilized blastocyst.

The most common therapy on men who suffered fertility decline due to aging was called “T Therapy”, but that’s therapy has long-term risks of sexual dysfunction, metabolic syndrome, prostate, and cardiovascular system. Stem cells are an alternative therapy can be used for ameliorate testicular fuction because of their ability to differentiate into various cell types. The aim of this study was to evaluate the injection of hWJ-MSC in physiologic aging male rats on testicular function. This study was used 3 young male rats (8-12 weeks) and 6 physiological aging male rats (22-24 months) which divided into 3 groups, (i) the young rats, (ii) physiological aging male rats, and (iii) physiological aging male rats that injected with hWJ-MSCs. The young rat group did not give any treatment, physiological aging male rats received NaCl (0.9%) 0.4 mL, and the treatment group received 1x106 cells/kg BW of hWJ-MSCs. The observations were performed on the macroscopical and histological analysis. The result indicates that the younger group had the lowest body weight (154.6 g) and the percentage of the testis weight on the body weight was highest (2.2%) compared to the other groups (P>0.05). The physiological aging rats group had the smallest tubule (9726.9 ìm2) with a largest interstitial area (1117.1 ìm2) compared the other groups (P>0.05). After injection of hWJ-MSC, the tubule area became wider followed by narrowing of the interstitial area (P>0.05). The difference in the body weights is due to the different age of the rat. Improvement of tubule area and interstitial area due to the ability of hWJ-MSCs to improve spermatogenic cells within the tubule. Injection of hWJ-MSCs has been shown to increase fertility in aging rats.

Oocyte vitrification is the important part of gamete preservation for further purpose. The objective of this study was to evaluate the response or development of vitrified-mice oocyte following activation using various concentrations of Strontium Chloride (Sr Cl2). Oocytes were collected from superovulation-induced female mice. Oocytes vitrification was then performed using a gradual equilibration of 2 M Ethylene Glycol in 0.25 M sucrose and 7 M Ethylene Glycol on 0.5 sucrose. Subsequently, the vitrified oocytes were thawed and activated using various Strontium Chloride concentration in each group. Control 1 is unvitrified oocyte and without Sr Cl2. Control 2 is unvitrified oocyte then activated by 20 mM Sr Cl2. Zero (0) mM Sr Cl2 is vitrified oocyte without Sr Cl2. Group Ten (10) mM is vitrified oocyte then activated by 10 mM Sr Cl2. Group Twenty (20) mM is vitrified oocyte then activated by 20 mM Sr Cl2. The viability of vitrified-thawed oocytes was observed based on ooplasm integrity. Whereas the oocytes respond to artificial activation was observed based on pronucleus formation after 10 hours of activation. The result showed that 39% of oocyte degenerated following vitrification. The respond of vitrified-thawed oocytes following artificial activation using Strontium Chloride was significantly lower compared to fresh oocytes (p

Stem cell technology has great potential in the effort to cure degenerative diseases. This study was done to determine optimum dose of flamboyant (Delonix regia) flower extract to induce proliferation and differentiation of mice (Mus musculus) bone marrow mesenchymal-like stem cell. Bone marrow cells were collected from mice by aspiration. Cells suspension (1 x 106) were poured into petri dishes containing 2 ml of modified Dulbecco's Modified Eagle's Media (mDMEM) and incubated overnight at 37 °C in a 5% CO2 incubator and microscopically observed. In quadriplicate, MSC were cultured in mDMEM containing D. regia flower extract of 0.0 (control), 0.4, 0.6, 0.8, and 1.0 mg/ml and incubated at 37 °C for 9 days. Population doubling time (PDT) and differentiated cell type were microscopically observed using HE staining on day 1 and 10. Data obtained were analyzed by ANOVA and Tukey test. The results showed that the addition of D regia flowers extracts 0.8 and 1.0 mg/ml significantly reduced PDT compared to that of 0.4, 0.6 and control. The extract, at 0.4 and 0.6 mg/ml, were able to induce MSC differentiation into fibroblast-like and nerve-like cells. In conclusion, D. regia flower extracts of 0.6, 0.8 and 1.0 mg/ml were able to stimulate MSC proliferation, but optimum dose for neurogenic differentiation was 0.6 mg/ml. This is the first to show potential of D. regia flower extract as neurogenic differentiatian inducer on mice MSC. These findings can be used as preliminary information for using the extract as cellular differentian inducer in basic and applicative reseach using stem cells.

Latar belakang. Pneumonia masih menjadi salah satu penyebab utama morbiditas dan mortalitas anak balita di seluruh dunia. Red-cell distribution width (RDW) dan mean platelet volume (MPV) adalah dua parameter dalam pemeriksaan darah lengkap yang sederhana, murah, dan tersedia hampir di seluruh rumah sakit di Indonesia. Penggunaannya sebagai biomarker keparahan pneumonia dapat menjadi salah satu upaya untuk menurunkan morbiditas dan mortalitas akibat pneumonia. Tujuan. Menganalisis hubungan antara nilai RDW dan MPV dengan derajat keparahan pneumonia anak. Metode. Penelitian cross-sectional dilaksanakan pada Juni-November 2017 di RSUD Nganjuk, melibatkan 30 anak berusia 2-59 bulan dengan pneumonia. Nilai RDW dan MPV dihubungkan dengan klasifikasi diagnosis, derajat keparahan yang diukur menggunakan skor RISC (respiratory index of severity in children) dan PRESS (pediatric respiratory severity score), dan hasil terapi pneumonia. Hasil. Terdapat perbedaan bermakna pada nilai RDW antara subjek dengan pneumonia dan pneumonia berat (t28=-1,721, p=0,096). Nilai RDW antara kelompok dengan nilai PRESS rendah, sedang, dan tinggi juga berbeda bermakna (F(2,27)=4,083, p=0,028). Nilai RDW berhubungan bermakna kuat dengan klasifikasi diagnosis pneumonia (x(1)=0,530, p=0,004), dan berhubungan bermakna moderate dengan skor RISC (rs=0,369, p=0,023), serta skor PRESS (rs=0,318, p=0,043). Kesimpulan. Nilai RDW berhubungan dengan klasifikasi diagnosis pneumonia, skor RISC dan skor PRESS.

Preservation of ovarian tissue from severely injured or dead valuable animals has the potential to preserve female germ cells of animals. The ability to mature and fertilize of oocytes from preserved ovary of endangered species will allow us to sustain genetic and global biodiversities. The aims of this study were to investigate the viability of oocytes collected from the preserved ovary and its potential utilization for the production of cat embryos followed by in vitro maturation and fertilization. Ovary was preserved immediately in phosphate buffer saline (PBS) at 4 °C for 24 or 48 hours. The quality and viability of oocytes after the maturation process were identified microscopically using aceto-orcein staining. Biological function of the oocytes was evaluated by using in vitro culture technique for the maturation and fertilization rate in CR1aa medium culture. The results showed that the percentage of oocytes collected from preserved ovary for 24 and 48 hours that remained at the stage of metaphase-II were 29.4% and 21.9% respectively. Fertilization rates produced in the IVF using oocytes collected from ovary preserved for 24 or 48 hours were significantly lower (30%) than that of unpreserved control (36.7%). In conclusion, female germ cells of cat ovary preserved at 4 °C in PBS for 2 days were still viable for in vitro fertilization and thus can be utilized for in vitro production of cat embryos. Information obtained can be used as a basis of knowledge of using a combination of physiological reagent and cold-based preservation technique in modern reproductive technology for animals.

Background: Epididymal sperm preservation is a simple conservation approach that can help prevent the loss of high genetic quality of farm animals. The chance of loss increases, especially during disease outbreaks or other interruptions to normal reproduction function. Aim: This study looked into the ability of preserved ram epididymal sperm to fertilize oocytes. Due to motility becoming an issue following sperm storage for fertilization, the sperm microinjection known as intracytoplasmic sperm injection approach was employed. Methods: The study was divided into two parts. First, involved the preservation of epididymal sperm at 5°C for 12 days. During preservation, sperm quality parameters namely motility, viability, intact membrane, acrosome, and Deoxyribonucleic acid (DNA) are evaluated every three days. For the fertility test in the second experiment, matured oocytes were injected with immotile sperm discovered in the last days of preservation. The presence of pronucleus development following in vitro culture is used as an indicator of sperm's ability to activate and fertilize oocytes. Results: All sperm quality parameters significantly (p < 0.05) declined during preservation time. On day 12, motility was discovered to be 0%, but viable sperm, sperm with intact membrane, acrosome, and DNA remained at 41.86% ± 9.30%, 31.18% ± 5.15%, 21.88% ± 1.93%, and 33.35% ± 8.74%, respectively. On the fertility test, we inject immotile sperm from day 12 of preservation, which has the lowest motility found, into matured oocytes. Those sperms are able to activate (52.05% ± 7.15%) and fertilize (31.37% ± 1.75%) the injected oocytes, but their fertilizing ability is significantly lower (p < 0.05) when compared to the sperm derived from the ejaculate. Conclusion: In this study, simple preservation of epididymal sperm reduces all sperm quality criteria, particularly motility. Using the microinjection approach preserved sperm which had no motility, still demonstrated its ability to activate and fertilize the oocytes. According to that, this study provides potential approaches and tools for using genetically superior animals that have lost their ability to execute regular fertilization, and also prolong reproduction function. [ABSTRACT FROM AUTHOR]

Conjunctival graft has been frequently used in the field of ophthalmology. The frequently used methods to attach a conjunctival graft are suture technique and the use of fibrin glue. This study was to investigate albumin glue as an alternative to suture technique in attaching conjunctival grafts in rabbits. The aim of this study was to compare the conjunctival wound healing between albumin glue and suture technique in rabbit eye as a model. This was an experimental animal study that included 32 eyes (16 rabbits) conducted at PT. Bio Farma (Persero) and the Histology Laboratory, Faculty of Medicine, Universitas Padjadjaran from March 2014 to July 2104. The subjects in this study were divided into albumin glue group and suture technique group. The examinations were comparison of conjunctival graft attachment and histologic microscopic examination to assess the wound gap. Data analysis was performed statistically using Mann-Whitney test for small sample. The statistical analysis results showed that the graft attachment was significantly better when using albumin glue (grade 4) compared to suture (grade 2–3) on day-1 after surgery (p=0.000). The wound gap was smaller using albumin glue (0-0,33 µm versus 5,33-14 µm; p0.0005) 10 minutes after surgery and 0 µm versus 0.33–4 µm, p 0,0005, on day-7 after surgery. In conclusion, graft attachment using albumi n glue is better and the wound gap is smaller when using albumin glue compared to the suture technique. [MKB. 2016;48(4):241–8]

The aim of this study was to determine the best combination of cryoprotectant (Ethylene glycol, EG), dimethyl sulfoxide (DMSO) and propanediol (PrOH) and carrier system (hemistraw and cryotop) in improving ewe oocytes viability during cryopreservation. Oocytes with multi layers of compact cumulus cells were colleted from abbatoir and matured in TCM 199 medium supplemented with 10% fetal calf serum for 24-26 h at 38.5° C under 5% CO2 in the air. Matured oocyte was divided into six parts and vitrified in three different vitrification solutions; (i) 17% EG+17% DMSO with hemistraw as carrier system, (ii) 34% EG with hemistraw as carrier system, (iii) 17% EG+17% PrOH in hemistraw (iv), 17% EG+17% DMSO with cryotop as carrier system (v), 34% EG with cryotop as carrier system (vi), and 17% EG+17% PrOH in cryotop. Oocytes were cryopreserved for one week before revived and evaluated for viability. The result showed that oocytes vitrified in media containing EG and DMSO in cryotop had the highest viability (88.16%) compared to media containing EG only or EG and PrOH (70.95% and 68.76%, respectively) (P0.05). Moreover, oocytes viability that vitrified using cryotop and hemistraw as carrier system were not significantly different. The present results indicated that vitrification using combination of EG and DMSO as permeable cryoprotectant and cryotop as carrier system was the best system to maintain oocyte viability after vitrification-warming.

A stable repaired fracture is the key factor responsible for the recovery of a damaged bone. The iron-based implant is one of the biodegradable metals that have been proven safe as a fracture fixation device. The objective of our experimental approach was to examine the potential of the iron-based implant as a biodegradable metal in tibia shaft fracture in sheep chronically. The samples used for this experiment were iron-based and stainless steel implants. Each had a diameter of 5 mm. These samples were analyzed through 3 phases which are material characterization, in vitro and in vivo examination. The samples were examined using a scanning electron microscope with energy dispersive spectrometer and X-ray diffraction. Based on the analysis carried out, the samples contained 90,02% and 60,81% Fe for iron-based implant and stainless implant, respectively. Both implants maintained high viability when being in contact with calf pulmonary artery endothelial cells, indicating that both implants had a minimum response to the cell in a hemocytometer and methyl tetrazolium (MTT) assay. The systemic effect of the implants was observed using hematology and blood chemistry examination. Data collection also shows that both implants also had a minimum response to the erythrocytes, leucocytes, blood chemistry, and blood mineral during the period of observation. Therefore, it could be concluded that the iron-based implant is tolerable for a period of 9 months. It also has the potential to be used as a biodegradable orthopedic implant.