We present a case of a 30-year-old man who committed suicide, using three knives, after a domestic disturbance in which he repeatedly stabbed his wife. During the inspection of the corpse of the man at the scene, two knives were found embedded in the neck and one in the abdomen. During the autopsy, we found two stab-incised injuries on the anterior surface of the neck in the lower section of the second segment bilaterally, one injury on the left side of the chest, and eleven injuries in the abdomen of the deceased with varying depth. Wound channels ranged from 1.5 cm to 20 cm. Despite our expectations that some of the major arterial and/or venous vessels in the neck would be cut, we found breaks in only small vessels, nervous structures, sublingual glands, and muscles in the course of the wound channels. In this case, we define such an autopsy finding as rare, or rather, as a rare accident. The inflicted stab-incised injuries as individual anatomical lesions would not lead to certain death. At the time of autopsy we revealed that the fatal injury was in the abdomen, wherein the abdominal aorta was cut. This led to the development of acute blood loss. In the genesis of death, a puncture-incised injury of the left lung was involved, leading to the development of hemopneumothorax. With regards to the possibility of murder, we did not find any cuts or puncture-incised injuries on the body of the deceased man which could be described as "defensive", or such that could have been received while trying to escape. During the police investigation, evidence for the presence of a third person at this domestic incident was not found.
Study question What is the qualitative and quantitative profile of microRNAs (miR) and their sequence variants - isomiRs, and how it changes during the menstrual-cycle phase transitions? Summary answer Time-course analysis of endometrial miR/isomiR profiles has shown that menstrual-phase transitions cause widespread and complex changes in miR gene expression and processing. What is known already Embryo implantation depends on the receptivity of the endometrium during the window of implantation, when ovarian hormones and genetic factors coordinate the development of the uterine lining and prepare it for embryo implantation. The most important factors for successful implantation studied so far are the embryo itself, the histological dating of the endometrium and its molecular genetic characteristics, including miRs. With the rapid development of next-generation sequencing technologies, it has become clear that miR genes have the potential to produce not only miR but also variants (isomiRs) thereof, which can differ in sequence and length and can be functionally significant. Study design, size, duration miR/isomiR landscape was assessed by small RNA sequencing of endometrial biopsy samples at 4 time points of endometrial cycle covering the proliferative and secretory phases. Healthy, fertile, female volunteers took part in the study lasting one and a half years. For accurate phase dating, human chorionic gonadotropin (hCG) was administrated, and ultrasonic, histological and hormonal assessments were done at each time point. Statistically significant data of miR/isomiR identification and expression dynamics was considered for analysis. Participants/materials, setting, methods Participant choice criteria - at least one child born, problem-free pregnancies, no diseases or allergies; hCG application time determined according follicle and endometrium ultrasound scanning, and ovarian hormone levels; endometrial biopsies taken at hGC (before hormone application), hGC+2, hGC+7, hGC+9 time points; small RNAseq completed by Karolinska Institute, Sweden; miR/isomiR identified using local Galaxy instance with an in-built workflow and tools developed by our laboratory; differential expression and target prediction evaluated with DESeq2 and miRDB,resp. Main results and the role of chance Within the cohort of patients, across the four study time points, the small RNAseq data revealed numbers of miRs and isomiRs to be changed. The largest statistically significant changes in their expression were found at LH + 9. The miR families that showed the largest number of members with altered expression were miR125a, miR30d, miR449c, miR92a/b and miR99a. The expression levels tended to decrease in the miR125a and miR92a families and to increase in the miR10a and miR449c families during the three studied time points of the cycle compared to the proliferative phase. Among those affected, the number of isomiRs, including templated and non-templated isomiRs, was much higher than that of miRs. For example, the ratio of the significantly altered miRs/templated isomiRs/non-templated isomiRs was 6/16/11 at LH + 9. Templated isomiRs of hsa-miR–148a–3p, hsa-miR–30d–5p and hsa-miR–449c–5p were among the most upregulated, while several templated and non-templated isomiRs of hsa-miR–125–5p were the most downregulated at LH + 9. Of particular interest are those isomiRs, in which the seed site is shifted compared to the reference miRs and results in altered target transcripts. Target prediction of the most affected isomiR of hsa-miR–449c–5p identified new targets of target scores much higher than of the reference miR. Limitations, reasons for caution IsomiRs are a source of novel biomarkers for clinical diagnosis. An important next step is the validation of the in-silico predicted miRs/isomiRs and their target transcripts by RT-qPCR in larger number of individuals. Expression profiles should be associated with the dominant cell type in the endometrial biopsy preparation. Wider implications of the findings: MiR/isomiR signatures, together with those of their target mRNAs, can be applied to distinguish the endometrial phases, especially the implantation window, as well as for diagnosing endometrial dysfunction. It is worth investigating the possibility of miRs/isomiRs being used as biomarkers not only in endometrial biopsy but also in liquid biopsy. Trial registration number The Bulgarian National Science Fund КП–06 Н31/2
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