@article {2294, title = {The Effect of Cryopreservation on the Sperm Ultrastructure of Mus Musculus Albinus Strain DDY: Comparison of Nakagata vs Modified vs Kitazato Cryoprotectants}, journal = {Pharmacognosy Journal}, volume = {16}, year = {2024}, month = {June 2024}, pages = {563-569}, type = {Original Article}, chapter = {563}, abstract = {

Introduction: Sperm morphology analysis is very necessary to understand male fertility and the etiology of infertility. Currently, scanning electron microscopy (SEM) has been widely used to determine surface topology. In this study, we will compare the effects of spermatozoa cryopreservation using three different types of cryoprotectants, namely Nakagata, modification and Kitazato. The cryoprotectant compositions used are Nakagata (raffinose and skim milk), modified (glycerol and raffinose) and Kitazato (glycerol and trehalose). Methods: SEM analysis was carried out on 8 sperm samples before cryopreservation and after the freeze-thaw process. Results: The results obtain showed that cryoprotectant modification was able to protect spermatozoa morphology better than Nakagata and Kitazato. Analysis revealed damage to plasma membrane, acrosome and loss of mitochondria in all treatment groups compared to fresh sperm. SEM showed obvious signs of post-thaw damage such as missing plasma membranes, sperm showing damaged acrosomes and mitochondria in the middle showing structural disorganization. Conclusion: SEM revealed that cryopreservation caused ultrastructural damage to mice sperm due to freezing and thawing. These details provide valuable data for further research to minimize the damage caused by cryopreservation to mice sperm. Apart from that, further examination using TEM is recommended to obtain a more comprehensive picture.

}, keywords = {cryopreservation, glycerol, mice sperm, raffinose, Scanning electron microscope, Trehalose}, doi = {10.5530/pj.2024.16.88}, author = {Manggiasih Dwiayu Larasati and Silvia W. Lestari and Mulyoto Pangestu and Andon Hestiantoro and Kusmardi Kusmardi} } @article {2293, title = {Effect of Glutathione Supplementation in Cryoprotectant Modification on Tyrosine Phosphorylation, Acrosin Expression and Acrosome Reaction of Post-Thawing Spermatozoa Quality}, journal = {Pharmacognosy Journal}, volume = {16}, year = {2024}, month = {June 2024}, pages = {554-562}, type = {Original Article}, chapter = {554}, abstract = {

Background: Tyrosine phosphorylation, acrosin, and acrosome reaction play an important role in fertilisation. However, cryopreservation causes changes in tyrosine phosphorylation, acrosin expression, and acrosome reaction which affect the quality of spermatozoa. Cryoprotectant media added with antioxidants is needed to protect Spermatozoa from the effects of cryopreservation so that the quality of spermatozoa can be maintained. Objectives: This research examined the effect of glutathione (GSH) supplementation in cryopreservation media on tyrosine phosphorylation, acrosin expression, and acrosome reaction. In this research, pure modified Cryoprotectant (CPA) was compared with CPA supplemented with GSH in three different concentrations. Materials and Methods: The research sample was male mus musculus albinus strain Deutchland Denken Yoken (DDY). Mice spermatozoa was cryopreserved and several parameters were measured including tyrosine phosphorylation, acrosin expression, and acrosome reaction. Results: The addition of GSH to the modified CPA increased tyrosine phosphorylation, acrosin expression, and acrosome reaction (maintaining acrosome integrity). The group with 1.00 mM GSH obtained the highest results among the other groups. Significant increases were found in tyrosine phosphorylation, acrosin expression, and acrosome reaction after the addition of 1.00 mM GSH. Conclusion: Glutathione supplementation in modified CPA can increase tyrosine phosphorylation, acrosin expression, and acrosome reaction of frozen-thawed spermatozoa. Treatment using GSH at a dose of 1.00 mM is the most effective and modification of CPA with the addition of glutathione can improve the tyrosine phosphorylation, acrosin expression and acrosome reaction in cryopreserved spermatozoa.

}, keywords = {acrosin expression, acrosome reaction, CPA modification, glutathione (GSH), spermatozoa cryopreservation, tyrosine phosphorylation}, doi = {10.5530/pj.2024.16.87}, author = {Zuraida and Silvia Werdhy Lestari and Mulyoto Pangestu and Andon Hestiantoro and Kusmardi Kusmardi} } @article {2202, title = {Inhibitory Mechanisms of Soybean Extract on the Development of Breast Cancer Through Modulation of Cellular Immune Response}, journal = {Pharmacognosy Journal}, volume = {16}, year = {2024}, month = {February 2024}, pages = {01-08}, type = {Original Article}, chapter = {01}, abstract = {

Background: Breast cancer is the most frequently diagnosed cancer in women worldwide. Consumption of soy products has been reported to reduce the incidence of and mortality rate for some cancers, including breast cancer. However, there are limited in vivo studies on the inhibitory effect of soybean extracts on breast cancer. Objectives: To examine the effect of soybean extracts on breast cancer cellular immunity and to determine the role of CD4+ and CD8+ T cells in the development and outcome of breast cancer. Material and Methods: Rat were induced with DMBA 11 times to get a breast cancer model. A soybean extract was given at different doses starting one week before DMBA induction and continued until the end of the study. At the end of the study, peripheral blood was collected, and the lymphocytes were examined using flow cytometry. Results: The phytochemical screening of soybean extract, using the Q-TOF LC/MS method, detected four bioactive components from the isoflavone and saponin groups. The incidence of tumor formation in the NeC, SE-D250, SE-D500, and SE-D1000 groups was 100\%, 83\%, 33\%, and 33\%, respectively. The highest proportion of CD4+ T cells was found in the NeC (69.35\%), while the lowest was in the SE-D1000 (63.75\%). The highest and lowest proportions of CD8+ T cells were found in the SE-D1000 and NeC groups, at 35.95\% and 31.15\%, respectively. Conclusions: The soybean extract was able to reduce the incidence of breast tumor formation in DMBA-induced rat in a dose-dependent manner. The soy extract group{\textquoteright}s CD4+/CD8+ ratio was close to that of healthy rats compared to the DMBA-induced group without soy extract. A lowered CD4+/CD8+ ratio is followed by a lower risk of tumor formation.

}, keywords = {Breast cancer, CD4+, CD8+, Cellular immune response, Soybean extract}, doi = {10.5530/pj.2024.16.1}, author = {Andi Muh. Maulana and Kusmardi Kusmardi and Erni Hernawati Purwaningsih and Andon Hestiantoro and Taifo Mahmud and Heri Wibowo and Bambang Pontjo Priosoeryanto and Primariadewi Rustamadji and Numlil Khaira Rusdi} } @article {1657, title = {In Vivo Antimammary Tumor Effects of Soybean Extract with Targeted Lunasin (ET-Lun)}, journal = {Pharmacognosy Journal}, volume = {13}, year = {2021}, month = {September 2021}, pages = {1269-1276}, type = {Research Article}, chapter = {1269}, abstract = {

Background/Objective: Lunasin is a peptide, consist of 44 amino acids which have anti-cancer, antioxidant, and anti-inflammatory activity. The price of commercial Lunasin was very expensive due to the high cost of lunasin synthesis and the lack of methods to obtain the pure lunasin weight from plant sources, involving time-consuming analytical instruments. To overcome these problems, the soybean extract with targeted Lunasin (ET-Lun) was made. The aim of this study was to investigate anti-cancer properties of ET-Lun in breast cancer models in vivo. Methods: Effect of ET-Lun was evaluated in 7,12-Dimetilbenz[a]antrasen (DMBA) induced breast cancer rat model. Tumor Mass, volume, and number were measured. The expression of HER2 and EGFR from each treatment group in DMBA-induced rat was evaluated using immunohistochemistry. Results: The results shown that ET-Lun could reduced tumor volume (p=0,021). ET-Lun decreased EGFR expression compared to negative control DMBA (p=0,012). Conclusions: These results indicated that the ET-Lun has anti-breast cancer activity in vivo.

}, keywords = {Breast cancer, EGFR, HER2, In-vivo, Soybean}, doi = {10.5530/pj.2021.13.160}, author = {Numlil Khaira Rusdi and Erni Hernawati Purwaningsih and Andon Hestiantoro and Berna Elya and Kusmardi Kusmardi} } @article {1673, title = {Subchronic Toxicity of Lunasin Targeted Extract (ET-Lun) from Soybean Seed (Glycine max (L.) Merr.): Perspective from Liver Histopathology, SGOT, and SGPT Levels in Sprague Dawley Rats}, journal = {Pharmacognosy Journal}, volume = {13}, year = {2021}, month = {November 2021}, pages = {1384-1388}, type = {Original Article}, chapter = {1384}, abstract = {

Background: Lunasin Targeted Extract (ET-Lun) has a pharmacology effect in inhibiting inflammation by decreasing COX-2 and iNOS expression. ET-Lun could increase apoptosis and decrease dysplasia (p \> 0,05). In addition, ET-Lun could decrease EGFR expression in breast cancer rats. The acute toxicity showed ET-Lun has LD50 more than 5000 mg/kg BW and was practically non-toxic. Objective: this study aimed to determine the subchronic toxicity of ET-Lun. Methods: Male and female Sprague Dawley rats (n=40) were divided into 4 groups, the control group and treatment group ET-Lun dose of 250 mg/Kg BW, 500 mg/kg BW, and 750 mg/kg BW. The ET-Lun was administered for 90 days. On the 91st day, the animals were dissected and examined for SGOT-SGPT levels, liver histopathology, and diameter of the central vein. Results: The SGOT-SGPT levels showed no significant difference between the treatment group and the control group (p \> 0.05). On microscopic observation, there was no change or damage to the liver of rats in each group. The diameter of the central vein of the rat liver shows no significant difference between the control and treatment groups. Conclusion: The ET-Lun does not produce adverse effects in liver rats after subchronic treatment.

}, keywords = {Liver, Lunasin, SGOT, SGPT, Soybean, Subchronic Toxicity}, doi = {10.5530/pj.2021.13.175}, author = {Numlil Khaira Rusdi and Weri Lia Yuliana and Erni Hernawati Purwaningsih and Andon Hestiantoro and Kusmardi Kusmardi} }