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"Kasus kekerasan seksual dalam bentuk persetubuhan dapat terjadi pada perempuan usia reproduksi yang sedang mengalami keputihan. Sekitar 75-80% dari semua perempuan setidaknya sekali menderita infeksi kandidiasis vulvovaginal dan infeksi tersebut menyumbang lebih dari 25% vaginitis menular. Kondisi keputihan akibat kandidiasis disebabkan oleh perubahan mikrobioma vagina dapat menimbulkan perubahan pH dan jumlah neutrofil vagina yang memengaruhi motilitas dan persistensi spermatozoa dalam vagina. Lama motilitas dan persistensi spermatozoa dalam vagina baik pada kondisi fisiologis maupun patologis merupakan dasar pertimbangan bagi praktisi forensik dalam penentuan waktu terjadinya persetubuhan. Penelitian ini bertujuan untuk menganalisis pH, jumlah neutrofil, dan profil mikrobioma vagina terhadap motilitas dan persistensi spermatozoa dalam vagina tikus kandidiasis. Penelitian eksperimental ini menggunakan tikus putih (Rattus norvegicus) galur wistar dengan tikus jantan sebanyak 6 ekor untuk sampel spermatozoa dan tikus betina sebanyak 32 ekor untuk perlakuan. Tikus betina dibagi dua kelompok yaitu tikus normal dan tikus kandidiasis. Pada kedua kelompok tikus betina diberikan perlakuan berupa inseminasi semen dalam vagina. Variabel yang diukur adalah pH, jumlah neutrofil, profil mikrobioma, motilitas dan persistensi spermatozoa dalam vagina. Analisis statistik perbedaan pH, jumlah neutrofil vagina, diversitas alfa mikrobioma, motilitas dan persistensi spermatozoa antara tikus kondisi normal dan model kandidiasis dengan uji Mann-Whitney, sedangkan perbedaan diversitas beta mikrobioma dilakukan uji permutational multivariate analysis of variance (Permanova). Estimasi survival motilitas dan persistensi spermatozoa dalam vagina menggunakan kurva kesintasan Kaplan-Meier dengan uji log-rank untuk penilaian signifikansi. Hubungan antara pH, jumlah neutrofil dan profil mikrobioma vagina dengan motilitas dan persistensi spermatozoa, dengan uji korelasi Spearman dilanjutkan uji regresi linear berganda. Hipotesis terbukti apabila pada uji perbedaan, uji log-rank,dan uji korelasi didapatkan nilai signifikansi p<0,05. Hasil penelitian menunjukkan terdapat perbedaan pH, jumlah neutrofil, motilitas dan persistensi spermatozoa antara kelompok tikus normal dengan tikus kandidiasis, namun tidak terdapat perbedaan profil mikrobioma tikus normal dan tikus kandidiasis. Kesintasan motilitas dan persistensi spermatozoa dalam vagina lebih lama pada tikus normal dibandingkan tikus kandidiasis. Terdapat korelasi antara pH dan jumlah neutrofil dengan motilitas dan persistensi spermatozoa dalam vagina tikus, namun tidak terdapat korelasi profil mikrobioma dengan motilitas dan persistensi spermatozoa dalam vagina tikus normal dan tikus kandidiasis. Semakin tinggi pH dan jumlah neutrofil dalam vagina, maka motilitas dan persistensi spermatozoa dalam vagina akan semakin menurun. Prosedur tambahan baru pada pemeriksaan kasus kekerasan seksual yang disarankan untuk dilakukan adalah pemeriksaan pH dan jumlah neutrofil vagina.

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Cases of sexual violence in the form of sexual intercourse can occur in women of reproductive age who are experiencing vaginal discharge. Approximate 75-80% of all women suffer from vulvovaginal candidiasis infection at least once and the infection accounts for more than 25% of infectious vaginitis. The condition of vaginal discharge due to candidiasis is caused by changes in the vaginal microbiome which can cause changes in vaginal pH and neutrophil count which affects the motility and persistence of spermatozoa in the vagina. Duration of motility and persistence of spermatozoa in the vagina, both physiological and pathological conditions is a basic consideration for forensic practitioners in determining time since intercourse. This study aimed to analyze pH, neutrophil count, and vaginal microbiome profile on the motility and persistence of spermatozoa in the vagina of candidiasis rat as a principles for determining the time since intercourse. An experimental study was conducted using white rats (Rattus norvegicus) Wistar strain with 6 male rats for spermatozoa samples and 32 female rats for treatment. Female rat were divided into two groups, namely normal and candidiasis rat. In both groups, female rat were given treatment in the form of vaginal insemination of semen. The variables measured included pH, neutrophil count, microbiome profile, motility and persistence of spermatozoa in the vagina. Statistical analysis of differences in pH, vaginal neutrophil count, microbiome alpha diversity, motility and persistence of spermatozoa between rat in normal conditions and candidiasis models was carried out using the Mann-Whitney test, while differences in microbiome beta diversity were carried out by the permutational multivariate analysis of variance (Permanova) test. Estimation of survival motility and persistence of spermatozoa in the vagina used the Kaplan-Meier survival curve with the log-rank test for significance assessment. The correlation between pH, neutrophil count and vaginal microbiome profile with spermatozoa motility and persistence, with the Spearman correlation test followed by multiple linear regression tests. The hypothesis is proven if the difference test, log-rank test and correlation test show a significance value of p<0.05.The results showed that there were differences in pH, neutrophil count, motility and persistence of spermatozoa between groups of normal and candidiasis rat, but there were no differences in the microbiome profiles of normal and candidiasis rat. The survival of motility and persistence of spermatozoa in the vagina is longer in normal rat than in candidiasis rat. There was a correlation between pH and the neutrophil count with the motility and persistence of spermatozoa in the vagina of rat, but there was no correlation between the microbiome profile and the motility and persistence of spermatozoa in the vagina of normal and candidiasis rat. The higher the pH and neutrophil count in the vagina, the motility and persistence of spermatozoa in the vagina will decrease. A new additional procedure in examining cases of sexual violence that is recommended to be carried out is checking the pH and vaginal neutrophil count."

"Latar Belakang: TNF-α dan CXCL16 terlibat dalam patofisiologi endometriosis melalui regulasi respon inflamasi dan pengkode nyeri endometriosis. Peningkatan TNF-α berperan dalam jalur pensinyalan P53 untuk apoptosis. Darah menstruasi sebagai pelepasan jaringan endometrium dapat digunakan dalam mengidentifikasi biomarker untuk diagnosis penyakit endometriosis tanpa memerlukan biopsi. Metode: Sampel darah menstruasi subjek dikumpulkan dengan menggunakan pembalut kertas saring dan jaringan endometrium dikumpulkan dengan melakukan biopsi, yang kemudian diekstraksi DNA dan RNA-nya. Tingkat metilasi DNA diukur dengan menggunakan metode pyrosequencing. Tingkat ekspresi mRNA diukur dengan menggunakan metode qPCR dan dianalisis dengan metode Livak Hasil: Ekspresi mRNA gen TNF-α pada darah menstruasi pasien endometriosis meningkat signifikan 3,73 kali lipat dibandingkan ekspresi pada kontrol (p=0,005). Gen TNF-α mengalami hipermetilasi dan berbeda bermakna dalam darah menstruasi pasien endometriosis dibandingkan kontrol (p=0,008). Sedangkan ekspresi mRNA gen CXCL16 pada darah menstruasi pasien endometriosis meningkat 2,42 kali (p=0,030) dibandingkan ekspresi mRNA darah menstruasi pada kontrol. Gen CXCL16 mengalami hipometilasi (p=0,004). Pada P53 terjadi terjadi peningkatan ekspresi gen P53 1,52 kali. Ekspresi mRNA gen TNF-α dan CXCL16 pada subjek nyeri berat lebih tinggi dibandingkan subjek nyeri sedang, dan terdapat korelasi positive. Kesimpulan: Penelitian ini menunjukkan bahwa peningkatan ekspresi mRNA TNF-α dan CXCL16 dalam darah menstruasi pasien endometriosis dapat menjadi penanda langsung untuk mendiagnosis endometriosis. Namun, untuk memvalidasi lebih lanjut temuan ini dan mengeksplorasi potensi sebagai alat diagnostik, penelitian tambahan yang melibatkan kelompok pasien yang lebih besar diperlukan

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Background: TNF-α and CXCL16 are implicated in the pathophysiology of endometriosis through the regulation of inflammatory response and the coding of endometriosis pain. Elevated TNF-α is implicated in the P53 signaling pathway for apoptosis. Menstrual blood, as a discharge of endometrial tissue, presents an opportunity for identifying biomarkers for the diagnosis of endometriosis without resorting to biopsy. Method: Menstrual blood samples were collected using filter paper pads, and endometrial tissues were obtained via biopsy, from which DNA and RNA were extracted. DNA methylation levels were assessed using the pyrosequencing method after bisulfite conversion treatment. Meanwhile, mRNA expression levels were measured using the quantitative polymerase chain reaction (qPCR) method and analyzed using the Livak method. Results: The mRNA expression of the TNF-α gene in menstrual blood of endometriosis patients increased significantly by 3.73 times compared to controls (p=0.005). The TNF-α gene exhibited hypermethylation, significantly differing in menstrual blood of endometriosis patients compared to controls (p=0.008). The mRNA expression of the CXCL16 gene in menstrual blood of endometriosis patients increased by 2.42 times (p=0.030) compared to controls, although there was no significant difference in expression between menstrual blood and endometrial tissue in endometriosis patients (p=0.173). The CXCL16 gene displayed hypomethylation (p=0.004). There was an increase in P53 gene expression, which was 1.52 times higher than in control menstrual blood. The mRNA expression of TNF-α and CXCL16 genes in subjects experiencing severe pain was higher than in those with moderate pain, and there was a positive correlation. Conclusion: This study suggests that increased mRNA expression of TNF-α and CXCL16 in menstrual blood of endometriosis patients may serve as direct markers for diagnosing endometriosis. However, further validation of these findings and exploration of their potential as diagnostic tools requires additional studies involving larger patient cohorts."