Analysis of MiR-20b, MIR-197 markers for differentiation between forensic body fluids encountered in sexual assault cases.

Identifying body fluids can be a critical clue that aids in reconstructing the crime scene. Semen and vaginal fluid identification is crucial, especially in cases of sexual assault. The majority of forensic studies focused on identifying normal body fluids and neglected the expression variation of semen in pathology. To differentiate between vaginal fluids, fertile and infertile semen samples (oligospermia and azoospermia) using miR 20b and miR197. A total of 48 body fluid samples, divided as 16 vaginal fluids, 16 fertile semen, and 16 infertile semen samples (8 with oligospermia and 8 with azoospermia), were collected, and the expression levels of miR-20b and miR-197 were detected by the SYBR Green real-time quantitative PCR technique. Our results showed significant different expression of these miRNAs in normal semen compared to vaginal and infertile semen. Moreover, we designed a model based on Fisher's discriminant function to forecast the group affiliations of unidentified samples. With three novel equations, we were able to accurately distinguish between semen and vaginal fluid, fertile and infertile semen, and oligospermia and azoospermia semen samples with validation accuracy of 81.3%, 100%, and 100%, respectively. MiR-20b and miR-197 expression levels are efficient and appropriate markers to distinguish semen from vaginal fluid and to differentiate between fertile and infertile semen samples. However, the present study is a preliminary study based on clinical samples, and the potential role of these markers in differentiating real crime scene samples is still unknown, so we recommend further research to investigate these markers expression while using forensic samples.

Potential role of the lncRNA "HOTAIR"/miRNA "206"/BDNF network in the alteration in expression of synaptic plasticity gene arc and BDNF level in sera of patients with heroin use disorder through the PI3K/AKT/mTOR pathway compared to the controls.

INTRODUCTION: Heroin use disorder (HUD) is a seriously increasing health issue, accounting for most deaths among drug abusers. Studying non-coding ribonucleic acid gene expression among drug abusers is a promising approach, as it may be used in diagnosis and therapeutics. PARTICIPANTS AND METHODS: A total of 49 male heroin-dependent patients and 49 male control participants were recruited from Kasr Al Ainy Psychiatry and Addiction outpatient clinics, Faculty of Medicine, Cairo University. Sera were gathered. qRT-PCR was utilized for the detection of gene expression of non-coding RNAs such as "HOX transcript antisense RNA" (HOTAIR), micro-RNA (miRNA-206), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), mechanistic target of rapamycin (mTOR), and Activity Regulated Cytoskeleton Associated Protein (Arc). Sera Brain-Derived Neurotrophic Factor (BDNF) levels were assessed using ELISA. Using a western blot made it possible to determine the protein expression of PI3K, AKT, and mTOR. RESULTS: The study demonstrated that gene expressions of HOTAIR, AKT, PI3K, and Arc were considerably lowered between cases and controls, while gene expressions of miR-206 and mTOR1 were significantly raised. PI3K and AKT protein expressions were downregulated, while mTOR expressions were upregulated. BDNF levels were significantly decreased in some cases. CONCLUSION: The results of this study suggest that decreased HOTAIR in HUD relieves miR-206 inhibition, which thus increases and affects downstream PI3K/AKT/mTOR, ARC, and BDNF expression. This may be shared in addictive and relapsing behaviors.

Lactobacillus DNA usage in differentiation of normal vaginal fluids in premenopausal and postmenopausal females.

The presence of vaginal fluid as a bio-stain in the crime scene of sexual assaults provides pivotal evidence. The vaginal secretions are known to be rich in Lactobacillus; hence the current work aims to identify vaginal secretions via detection and quantification of Lactobacillus DNA in pre and postmenopausal females and to test its stability over storage time using Critical Threshold method applied by Polymerase chain reaction approach. Comparative study is done by Critical Threshold and Relative Expression methods aiming to evaluate the two methods. Results showed that (ΔCT) <9 powerfully indicates the presence of vaginal fluids. Values of ΔCT in all vaginal samples are stable and not affected by storage. Two novel cutoff values are obtained in order to differentiate between premenopausal and postmenopausal vaginal fluid samples which are (8.42) using the Critical Threshold method and (0.24) using the Relative Expression method. One novel cutoff value is obtained to differentiate between fresh and stored vaginal samples by the Relative Expression method which is (0.39). It is concluded that Lactobacillus DNA quantification via PCR is a good positive identifier for vaginal secretions which is remarkably stable over storage time.