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All Journal Narra J
Puspita E. Wuyung
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Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Medicine & Pharmacology Public Health

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Effect of pH and neutrophil count on the motility and persistence of spermatozoa in the vagina of candidiasis rat models Iswara, Raja AW.; Hestiantoro, Andon; Budiningsih, Yuli; Werdhani, Retno A.; Birowo, Ponco; Wuyung, Puspita E.; Afandi, Dedi
Narra J Vol. 4 No. 2 (2024): August 2024
Publisher : Narra Sains Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.52225/narra.v4i2.823

Abstract

Sexual violence is a global issue affecting individuals regardless of their relationship to the perpetrator or the setting. Microscopic examination of spermatozoa from vaginal swabs is crucial in investigating cases of sexual intercourse to determine the time of the crime. Factors such as vaginal pH and neutrophil count influence the motility and persistence of spermatozoa in the vagina, particularly in conditions like candidiasis, highlighting the need for further research in this area. This study aimed to determine the effect of pH and neutrophil count on the motility and persistence of spermatozoa in the vagina with candidiasis. An experimental study was conducted using white rats (Rattus norvegicus) of the Wistar strain, with four male rats providing spermatozoa samples and 32 female rats receiving treatment. The female rats were divided into two groups: the normal group and the candidiasis model group. In both groups, the female rats were given vaginal insemination of spermatozoa. Variables measured included pH, neutrophil count, motility, and persistence of spermatozoa in the vagina. Data were analyzed using the Mann-Whitney test, followed by the Spearman correlation test. The findings revealed that spermatozoa motility lasted up to three minutes in normal rats, whereas in the candidiasis model, it was reduced to two minutes. Additionally, spermatozoa persistence in the vagina lasted up to six days in the normal group compared to up to three days in the candidiasis model. There were significant differences in pH, neutrophil count, motility, and persistence of spermatozoa in the vagina between the normal group and the candidiasis model (all had p<0.001). There was a correlation between pH and neutrophil count with the motility and persistence of spermatozoa in the rat’s vagina (p<0.001). In conclusion, vaginal pH and neutrophil count influence the motility and persistence of spermatozoa in the vagina of candidiasis rat models.
Dual sgRNA-directed knock out survivin gene expression using CRISPR/Cas9 technology for editing survivin gene in triple-negative breast cancer Syahrani, Resda A.; Wanandi, Septelia I.; Arumsari, Sekar; Nihayah, Silviatun; Watanabe, Yukihide; Mizuno, Seiya; Louisa, Melva; Wuyung, Puspita E.
Narra J Vol. 4 No. 3 (2024): December 2024
Publisher : Narra Sains Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.52225/narra.v4i3.1177

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease 9 (CRISPR/Cas9) offers a robust approach for genome manipulation, particularly in cancer therapy. Given its high expression in triple-negative breast cancer (TNBC), targeting survivin with CRISPR/Cas9 holds promise as a therapeutic strategy. The aim of this study was to design specific single guide ribonucleic acid (sgRNA) for CRISPR/Cas9 to permanently knock out the survivin gene, exploring its potential as a therapeutic approach in breast cancer while addressing potential off-target effects. Survivin gene knockout was conducted in the TNBC cell line BT549. Intron 1, exon 2, and intron 2 of the survivin gene were selected as sgRNA targets. These sgRNAs were designed in silico and then cloned into a CRISPR/Cas9 expression plasmid. The cleavage activity was assessed using an enhanced green fluorescent protein (EGFP) expression plasmid. The sgRNAs with higher cleavage activity were selected for the establishment of knockout cells. After transfecting the plasmid into the cells, the success of the survivin gene knockout was validated at the deoxyribonucleic acid (DNA) level using polymerase chain reaction (PCR) and sequencing analysis, and at the protein expression level using Western blotting. The study found that sgRNAs survin1A (targeting intron 1), survex2A (targeting intron 2), and survin2A (targeting intron 2) demonstrated higher cleavage activities compared to the other sgRNAs. However, using the single sgRNA, survex2A did not generate mutations in the survivin gene. At the protein level, survivin was still expressed, indicating that a single sgRNA was ineffective in knocking out the survivin gene. In contrast, the combination of sgRNA survin1A and sgRNA survin2A was more effective in generating mutations in the survivin gene, resulting in the deletion of the entire exon 2 and leading to a loss of survivin protein expression. In conclusion, our work provides specific sgRNAs and demonstrates the utilization of dual sgRNAs strategy in the CRISPR/Cas9 technology to knock out the survivin gene, showing potential in breast cancer therapy.
Impact of semen insemination on the vaginal microbiome profile of candidiasis rat model: A preliminary forensic study on sexual violence evidence Iswara, Raja AFW.; Hestiantoro, Andon; Budiningsih, Yuli; Werdhani, Retno A.; Birowo, Ponco; Wuyung, Puspita E.; Fadilah, Fadilah; Afandi, Dedi
Narra J Vol. 5 No. 1 (2025): April 2025
Publisher : Narra Sains Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.52225/narra.v5i1.1256

Abstract

Sexual violence, including sexual intercourse, can occur in women experiencing vaginal discharge, particularly in cases of vaginal candidiasis. In candidiasis, the vaginal microbiome undergoes changes that could serve as a diagnostic indicator or as evidence of sexual activity.  The aim of this study was to assess the effects of semen insemination on the vaginal microbiome profile of candidiasis rats and to determine its forensic investigations in cases of sexual violence.  An experimental study was carried out using Wistar strain rats (Rattus norvegicus), consisting of four male rats (for spermatozoa donors) and twenty-four female rats. The female rats were divided into four groups: normal condition (control), normal condition post-semen insemination, candidiasis rats, and candidiasis rat post-semen insemination. Vaginal microbiome profiles were examined for each group, using alpha diversity (Chao 1, Shannon, Simpson, and Faith PD indices) and beta diversity (Bray Curtis, Jaccard, Unweighted Unifrac and Weighted Unifrac indices). Data were analyzed using the Kruskal-Wallis test for alpha diversity and the PERMANOVA test for beta diversity. The vaginal microbiome profiles of normal and candidiasis rats showed no significant differences (p>0.05). In candidiasis rats, the microbiome predominantly consisted of the Eukaryota kingdom, particularly Candida albicans. Semen insemination did not significantly affect the vaginal microbiome profile of candidiasis rats in the short term (p>0.05). However, the study highlights that the presence of Eschericia-Shigella, Roseomonas, and Archaea in the vaginal microbiome post-semen insemination potentially serves as an indicator of infection or sexual activity in forensic contexts.
Co-Authors Alwahdy, Ahmad S. Andon Hestiantoro Berry Juliandi Dedi Afandi Dewi I.S. Santoso Erni Rahmawati Fadilah Fadilah, Fadilah Hedi R. Dewoto Iswara, Raja AFW. Iswara, Raja AW. Kurniawan, Mohammad Melva Louisa Mizuno, Seiya Muhammad I. Ilmiawan Nihayah, Silviatun Nurjati C. Siregar Ponco Birowo Ramli, Yetty Retno A. Werdhani SEKAR ARUMSARI, SEKAR Septelia I. Wanandi Susanto, Yayi D.B. Susy O. Lontoh Sutjahjo Endardjo Syahrani, Resda A. Watanabe, Yukihide Yuli Budiningsih