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Analysis AZF Gene Deletions in Infertile Men in Indonesia Hanizar, Evi; Hinting, Aucky
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

Show Abstract | Download Original | Original Source | Check in Google Scholar

Infertile men with the low quantity and quality of sperm associated with gene deletions in the long arm of chromosome Y (YQ), in the region known as the AZF region (Azoospermic Factor). This study aims to analyze the AZF gene deletions in infertile men with abnormal sperm categories ranging oligozoospermia, Oligoasthenoteratozoospermia until azoospermia. This study includes the type of cross-sectional observational study and DNA samples obtained from the blood of primary infertile men. Extraction of DNA uses a DNA extraction kit and DNA amplification uses polymerase chain reaction method. The analysis includes the number, motility and morphology of sperm conducted by WHO standard. Analysis of deletion is determined from the size of the base pairs of DNA amplification product. The results showed the prevalence of deletions in azoospermic category higher than the prevalence in other categories. Sequentially, the prevalence of deletions was followed by a severe category Oligoasthenoteratozoospermia, Oligoasthenoteratozoospermia, severe oligoteratozoospermia and oligoteratozoospermia. The most frequent gene deletions are sY86 gene, followed by sY84, while the most rare gene deletions are DAZ gene / sY255. This is in contrast with previous studies because of the background sample of infertile men from several races in Indonesia, the number of samples and the location of genes analyzed in the sub-region AZF. Deletions involving many genes in the AZF subregion associated with the smaller quantity and quality of sperm. Key Words : male infertility, azoospermic factor, gene deletion, sperm abnormality

Density Gradient Centrifugation Pra-freezing Mengoptimalkan Persentase Morfologi Normal Spermatozoa Pasca-thawing Suyono, Seso Sulijaya; Hinting, Aucky; Lunardhi, Hamdani; I'tishom, Reny
Majalah Kedokteran Bandung Vol 50, No 3 (2018)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Kriopreservasi akan mengganggu struktur dan fungsi spermatozoa. Preparasi semen mampu menghasilkan spermatozoa dengan kualitas baik. Penelitian ini bertujuan menganalisis pengaruh preparasi semen dengan density gradient centrifugation (DGC) pra-freezing terhadap kualitas spermatozoa pasca-thawing. Penelitian dilakukan di boratorium Biologi Kedokteran, Fakultas Kedokteran Universitas Airlangga periode November 2017â€“Januari 2018. Penelitian eksperimental laboratoris dilakukan menggunakan cairan ejakulat volunter pria infertil. Semua sampel dibagi menjadi dua bagian, kelompok kontrol serta kelompok perlakuan berupa preparasi mini DGC. Setelah penambahan krioprotektan, selanjutnya dilakukan freezing. Pemeriksaan kualitas spermatozoa, meliputi motilitas, viabilitas serta persentase morfologi normal menggunakan metode WHO 2010, baik pra-freezing maupun pasca-thawing. Persentase perubahan kualitas spermatozoa pasca-thawing kedua kelompok dibandingkan menggunakan uji t dan bermakna jika nilai p<0,05. Total 20 sampel cairan ejakulat digunakan dalam penelitian ini. Persentase penurunan motilitas progresif, motilitas total, serta viabilitas pasca-thawing antara kedua kelompok tidak berbeda bermakna dengan nilai p masing-masing 0,422; 0,873 serta 0,432. Namun, penurunan persentase morfologi normal pasca-thawing pada kelompok kontrol jauh lebih besar daripada kelompok perlakuan dengan nilai p<0,001. Penelitian ini menyimpulkan bahwa preparasi semen berupa mini DGC pra-freezing mampu menghasilkan spermatozoa pasca-thawing dengan persentase morfologi normal yang lebih baik daripada protokol direct freezing.Pre-freezing Density Gradient Centrifugation Optimizes the Percentage of Post-thawed Sperms with Normal MorphologyÂ Cryopreservation impairs sperm structure and functions. Sperm preparation is a selection technique to obtain a population of high quality sperms. This study aimed to analyze the effect of pre-freezing Density Gradient Centrifugation (DGC) sperm preparation on the quality of post-thawed sperms. An experimental laboratory study was conducted using the ejaculates collected from volunteers visiting to our department. Samples were split into two fractions: control group and treatment group. In the treated group, mini-DGC sperm preparations were developed. After some cryoprotectants were added, samples were then cryopreserved using rapid freezing protocol. The evaluation of sperm quality that included evaluation on motility, viability and percentage of sperm morphology was performed by referring to 2010 WHO standardization on semen analysis. Evaluation was performed under pre-freezing and post-thawed condition. The percentage of sperm parameter changes between the two groups were compared using t-test with p-value <0.05 considered statistically significant. A total of 20 samples were included in this study. Post-thawed progressive motility, total motility, and viability considerably declined between two groups with p-values of 0.422, 0.873, and 0.432 respectively. In post-thawed observation, the percentage of sperms with normal morphology was significantly lower in the control group when compared to the treatment groupÂ (p<0.001). In conclusion, the pre-freezing mini-DGC can optimize the post-thawed percentage of sperms with normal morphology compared to direct freezing protocol

Isolation and Identification of Transforming Growth Factor β from In Vitro Matured Cumulus Oocyte Complexes . WIDJIATI; ARIEF BOEDIONO; SUTIMAN BAMBANG SUMITRO; AUCKY HINTING; . AULANI’AM; TRINIL SUSILOWATI
HAYATI Journal of Biosciences Vol. 19 No. 1 (2012): March 2012
Publisher : Bogor Agricultural University, Indonesia

Transforming growth factor-β (TGF-β) is a two-chain polypeptide with molecular weight of 25 kDa which takes significant role in the steroidogenesis process. In the ovarian oocyte in particular, TGF-β has an important role in regulating reproductive function. TGF-β represents a key intrafollicular protein that regulates follicle development and aromatization process. The purpose of this research was to characterize and identify a protein fraction of TGF-β from the bovine isolated oocytes, which is synthesized during in vitro oocyte maturation process. Oocytes were collected from follicles with diameter of 3-8 mm. Oocytes were then matured in TCM 199 media supplemented with 5 μg/mg LH, 3% BSA, and 50 μg/ml gentamicin sulfate, and cultured in CO2 incubator (5%, 38.5 oC) for 20 hours. TGF-β receptors were identified immunohistochemically. Characteristics of the TGF-β protein were determined using SDS PAGE and TGF-β specification was tested using Western Blotting. The results showed that TGF-β receptors were identified and found in cumulus oocyte complexes (COCs). TGF-β protein was isolated from bovine oocytes with molecular weight 25 kDa and it was identified by Western blotting methods in the same molecular weight.

Antibody of goat zona pellucida-3 (gzp3) protein of mice(Mus musculus) block in vitro fertilization of mice as an animal model= Antibodi protein zona pelusida-3 kambing (gZP3) asal mencit(Mus musculus) mencegah ... Imam Mustofa; Laba Mahaputra; Yoes Priyatna Dachlan; Fedik Abdul Rantam; Suwarno .; Widjiati .; Aucky Hinting
Jurnal Sain Veteriner Vol 24, No 1 (2006): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

The researchs of immunocontraception have done in ZP3 of several species, but have not been done in ZP3 of goat. In preliminary study, gZP3 protein was effective prohibited of graviditation of mice. The aim of this study was to prove the potency of gZP3 protein to prohibit in vitro fertilization of mice as an animal model. Antibody of gZP3 produced on mice. Immunized mice serum was analyzed using Elisa and Dot blotting method. Antibody of gZP3 supplemented into M-16 media for oocyte incubation, continued with in vitro fertilization. The result showed that antibody titer of immunized mice serum was higher (p

Analysis AZF Gene Deletions in Infertile Men in Indonesia Evi Hanizar; Aucky Hinting
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UPT Penerbitan Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar

Infertile men with the low quantity and quality of sperm associated with gene deletions in the long arm of chromosome Y (YQ), in the region known as the AZF region (Azoospermic Factor). This study aims to analyze the AZF gene deletions in infertile men with abnormal sperm categories ranging oligozoospermia, Oligoasthenoteratozoospermia until azoospermia. This study includes the type of cross-sectional observational study and DNA samples obtained from the blood of primary infertile men. Extraction of DNA uses a DNA extraction kit and DNA amplification uses polymerase chain reaction method. The analysis includes the number, motility and morphology of sperm conducted by WHO standard. Analysis of deletion is determined from the size of the base pairs of DNA amplification product. The results showed the prevalence of deletions in azoospermic category higher than the prevalence in other categories. Sequentially, the prevalence of deletions was followed by a severe category Oligoasthenoteratozoospermia, Oligoasthenoteratozoospermia, severe oligoteratozoospermia and oligoteratozoospermia. The most frequent gene deletions are sY86 gene, followed by sY84, while the most rare gene deletions are DAZ gene / sY255. This is in contrast with previous studies because of the background sample of infertile men from several races in Indonesia, the number of samples and the location of genes analyzed in the sub-region AZF. Deletions involving many genes in the AZF subregion associated with the smaller quantity and quality of sperm. Key Words : male infertility, azoospermic factor, gene deletion, sperm abnormality

PENGARUH ESCHERICHIA COLI DAN GRANULOSIT TERHADAP KADAR REACTIVE OXYGEN SPECIES SECARA IN VITRO Sukarjati; Doddy M. Soebady; Aucky Hinting; Sudjarwo
JURNAL PENELITIAN BIOLOGI BERKALA PENELITIAN HAYATI Vol 15 No 2 (2010): June 2010
Publisher : The East Java Biological Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23869/268

In humans, male genital tract infection has been recognized as one of the causes of infertility. Indicators of the occurrence of genital tract infection are the presence of bacteria during semen culture (bacteriospermia) and the finding of leucocyte of more than 1 million/ml semen (leucotytospermia). Escherichia coli (E. coli) is the most common cause of prostatitis and epididymitis. The objective of this study was to determine the effects of E. coli and granulocytes on Reactive Oxygen Species (ROS) level in vitro. This study comprised of two experiments. In the experiment 1, sperm was incubated with E. coli, and the experiment 2, the sperm was incubated with granulocyte. In those experiments, ROS levels were observed. Spermatozoa were obtained from donor with normal spermatozoa according to WHO (1999). Escherichia coli was obtained by culturing the semen of infertile males. Granulocytes were obtained from donors’ blood. Sperm preparation was made by using Percoll gradient column method. Granulocyte isolation used Histopaque 1077 and 1119. ROS level was detected by means of chemiluminescence method with beta counter device. The result of this study showed that in vitro E. coli had the effect on ROS level, both stimulated by peroxidase (p = 0.000) and PMA (p = 0.006). Granulocyte had effect on ROS level. In peroxidase-stimulated ROS level, there was the effect between spermatozoa and granulocyte-incubated sperm (p = 0.000), granulocyte-incubated sperm and granulocyte (p = 0.002), and sperm and granulocyte (p = 0.000). In PMA-stimulated ROS level, there was effect between sperm and granulocyte-incubated sperm (p = 0.000), sperm and granulocyte (p = 0.000), granulocyte-incubated sperm and granulocyte (p = 0.000). In conclusion, under in vitro experiment, sperm incubated with E. coli and sperm incubated with granulocyte had the effect on the level of the ROS.

HUBUNGAN KADAR MDA SPERMA DENGAN INTEGRITAS MEMBRAN SPERMATOZOA TIKUS (Rattus norvegicus) SETELAH PEMAPARAN 2–METHOXYETHANOL Alfiah Hayati; Soesanto Mangkoewidjojo; Aucky Hinting; Sukarti Moeljopawiro
JURNAL PENELITIAN BIOLOGI BERKALA PENELITIAN HAYATI Vol 11 No 2 (2006): June 2006
Publisher : The East Java Biological Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23869/386

In the body, 2-methoxyethanol compound may be converted to MAA. MAA is a strong oxidant and may cause oxidation stress in spermatozoa. Oxidation stress is a disturbance on phosphorilation that increases ROS concentration, and it produces lipid peroxide in spermatozoa membrane resulted in high MDA concentration. One of indicator of spermatozoa membrane disturbances is a lack of spermatozoa membrane integrity. The main purpose of this research was to determine the relationship between MDA concentration in sperm and membrane integrity of spermatozoa in rats. The animal of treated groups (n = 40) were divide into 8 groups of 5 each. The rats were given subcutaneous injection with 0,2 ml of 200 mg/kg/day for 1 day (P1), 3 days (P2), 6 days/week (P3), and 12 days/two weeks (P4), respectively the control group was injected with physiological saline of the same volume. The concentration of MDA was measured by spectrophotometer and observing membrane integrity used HOS method to watch the spermatozoa response on hypoosmotic condition. The results of the research indicated that 2-ME caused the increasing in sperm MDA concentration and the decrease of spermatozoa membrane integrity. There was negative correlations between MDA concentration and spermatozoa membrane integrity.

THE ROLE OF OXIDATIVE PHOSPHORYLATION IN THE MECHANISM OF HUMAN SPERM MOTILITY REGULATION Sudjarwo Sudjarwo; Aucky Hinting; Noor Cholies Zaini
JURNAL PENELITIAN BIOLOGI BERKALA PENELITIAN HAYATI Vol 12 No 1 (2006): December 2006
Publisher : The East Java Biological Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23869/404

Energy synthesis for sperm motility in the mitochondria with enzymatic reactions in nucleus and mitochondria is called Oxidative Phosphorylation (OXPHOS), in which the respiratory process is coordinated in nucleus and mitochondria. Enzymes in OXPHOS process are complex enzymes I, III, IV and V in the mitochondria and complex enzyme II in the nucleus. If there is OXPHOS dysfunction, the mutation of mitochondria DNA (mtDNA) will decrease energy (ATP, Adenosine Triphosphate) production and increase Reactive Oxygen Species (ROS). The characteristics of ROS are oxidant for lipid, protein and DNA, and all these reactions lead to the dysfunction of OXPHOS and, in the spermatozoa, they will decrease motility. The detection of ROS, Malondialdehyde (MDA), 8-hydroxydeoxyguanosine (8-OH-dG), and sperm motility can use Chemiluminicense method, Spectroflourometric method, High Performance Liquid Chromatography (HPLC) method and Markler method, respectively, as World Health Organization (WHO) Procedures Laboratory. These results indicated there was significant indirect correlation between ROS with sperm motility and direct correlation between ROS with MDA, 8-OH-dG.

Interpretation of Detection SARS Cov-2 in Semen Cennikon Pakpahan; Agustinus Agustinus; Aucky Hinting
Indonesian Andrology and Biomedical Journal Vol. 1 No. 1 (2020): June
Publisher : Universitas Airlangga

Background: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a virus with a high virulence which cause SARS-CoV-2, a disease with potentially dangerous implications for human health and pandemic. The involvement of other organs in the spread of this virus is still being debated. Considering the presence of (Angiotensin Converting Enzyme-2 (ACE-2) and Transmembrane Serine Protease 2 (TMPRSS2) in the reproductive organs including male reproduction,the male reproductive system possiblity for spreading SARS-CoV-2 should be studied. Reviews: Five studies were reveal the presence of SARS-CoV2 in semen. The reported results are inconsistent. Some of these studies also used unclear methods and procedures, which led to bias in the final results. Ongoing studies are needed to confirm the definite findings before specific recommendations can be made for further management. Summary: There is no definite interpretation of whether SARS-CoV-2 spreads through semen, but protection is still needed when it comes into contact with the semen.

Technology Updates in Male Infertility Management Aucky Hinting; Agustinus Agustinus
Indonesian Andrology and Biomedical Journal Vol. 2 No. 2 (2021): December
Publisher : Universitas Airlangga

Background: Technologies are replacing manpower many fields including medical field. Several devices have been marketed for replacing/reducing manpower in the medical field including male infertility. Here we reviewed several technologies that developed in male infertility. Review: Computer assisted sperm analysis (CASA), Automatic assessment of biochemical marker of seminal plasma, B-mode ultrasound, and automatic sperm cryopreservation can be applied routinely. Several updates i.e. automatic histopathology assessment, ultrasound strain elastography, magnetic resonance imaging (MRI) stronger than 3.0 T, Artificial intelligence for predicting the presence of sperm in azoospermia cases, automatic sperm selection, and automatic intracytoplasmic sperm injection (ICSI) need more studies before their application. Summary: Prudent choice based on valid studies is needed in order to give a comprehensive management to patient with male infertility without using useless technology.

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