ABSTRACT
PURPOSE: After sexual assault there is a limited amount of time before the DNA evidence on the surface of the victim's body is not recoverable. During an assault, the offender may leave saliva on the victim's skin. Traditional examination methods use a swabbing technique to collect saliva for DNA testing. Victim activity, especially hygiene activity such as showering, may negatively affect DNA recovery. METHODS: In this experiment, we compared two techniques for recovery of salivary DNA from the skin's surface after a victim showers. We compared the traditional swabbing method to a "wet-vacuum" method using the M-Vac© to collect saliva from four body regions (neck, arm, stomach, and leg). In our research, we tested whether either collection technique obtained enough salivary DNA for autosomal and Y-STR analysis. In addition, we tested whether the M-Vac© is more effective at collecting DNA from large surface areas than traditional methods, by determining the amount of DNA collected. RESULTS: With both collection techniques, we were able to obtain male salivary DNA from at least one body region of the female after she had showered. There was no statistical difference in the amount of DNA collected between the swabbing technique and the M-Vac©. Autosomal STR analysis failed to detect the male contributor's DNA; therefore, we used Y-STRs. With Y-STR analysis, 47 samples returned a full male profile, and 26 samples returned a partial male profile after sample concentration. CONCLUSIONS: This research shows that salivary DNA can be collected from skin after showering and successfully analyzed using Y-STRs.
Subject(s)
Baths/methods , Chromosomes, Human, Y , Crime Victims , DNA/isolation & purification , Forensic Genetics/methods , Saliva/chemistry , Sex Offenses , Skin/chemistry , Specimen Handling/methods , Female , Genetic Markers , Humans , Male , VacuumABSTRACT
Type 1 diabetes is a polygenic disease that results in an autoimmune response directed against insulin-producing beta cells. PTPN2 is a known high-risk type 1 diabetes associated gene expressed in both immune- and pancreatic beta cells, but how genes affect the development of autoimmune diabetes is largely unknown. We employed CRISPR/Cas9 technology to generate a functional knockout of PTPN2 in human pluripotent stem cells (hPSC) followed by differentiating stem-cell-derived beta-like cells (sBC) and detailed phenotypical analyses. The differentiation efficiency of PTPN2 knockout (PTPN2 KO) sBC is comparable to wild-type (WT) control sBC. Global transcriptomics and protein assays revealed the increased expression of HLA Class I molecules in PTPN2 KO sBC at a steady state and upon exposure to proinflammatory culture conditions, indicating a potential for the increased immune recognition of human beta cells upon differential PTPN2 expression. sBC co-culture with autoreactive preproinsulin-reactive T cell transductants confirmed increased immune stimulations by PTPN2 KO sBC compared to WT sBC. Taken together, our results suggest that the dysregulation of PTPN2 expression in human beta cell may prime autoimmune T cell reactivity and thereby contribute to the development of type 1 diabetes.
Subject(s)
Diabetes Mellitus, Type 1 , Insulin-Secreting Cells , Pluripotent Stem Cells , Humans , Diabetes Mellitus, Type 1/genetics , T-Lymphocytes , Cell Differentiation , Protein Tyrosine Phosphatase, Non-Receptor Type 2/geneticsABSTRACT
Stem cell-derived ß-like cells (sBC) carry the promise of providing an abundant source of insulin-producing cells for use in cell replacement therapy for patients with diabetes, potentially allowing widespread implementation of a practical cure. To achieve their clinical promise, sBC need to function comparably with mature adult ß-cells, but as yet they display varying degrees of maturity. Indeed, detailed knowledge of the events resulting in human ß-cell maturation remains obscure. Here we show that sBC spontaneously self-enrich into discreet islet-like cap structures within in vitro cultures, independent of exogenous maturation conditions. Multiple complementary assays demonstrate that this process is accompanied by functional maturation of the self-enriched sBC (seBC); however, the seBC still contain distinct subpopulations displaying different maturation levels. Interestingly, the surface protein ENTPD3 (also known as nucleoside triphosphate diphosphohydrolase-3 [NDPTase3]) is a specific marker of the most mature seBC population and can be used for mature seBC identification and sorting. Our results illuminate critical aspects of in vitro sBC maturation and provide important insights toward developing functionally mature sBC for diabetes cell replacement therapy.
Subject(s)
Adenosine Triphosphatases/metabolism , Embryonic Stem Cells/metabolism , Induced Pluripotent Stem Cells/metabolism , Insulin-Secreting Cells/metabolism , Adenosine Triphosphatases/genetics , Calcium/metabolism , DNA, Mitochondrial , Gene Expression Regulation , Humans , TranscriptomeABSTRACT
BACKGROUND: The initial management of distal radius fractures in children is part of the usual practice of Emergency Medicine. However, no data are available evaluating the outcome of pediatric forearm fractures that undergo closed reduction and casting by emergency physicians. STUDY OBJECTIVE: To assess short-term outcomes after distal forearm fracture reductions performed by emergency physicians. METHODS: A retrospective cohort study with matched controls was performed on children with a closed, displaced, or angulated distal forearm fracture that required manipulation. The study group was defined as patients in whom emergency physicians performed closed manipulation and cast immobilization without orthopedic consultation. The control group was defined as patients who had closed reduction by an orthopedic resident. Two controls were identified for each study patient on or around the same date of visit. During the 20-month period, the medical records of 22 study patients and 42 controls were reviewed. The two groups were similar in age, fracture angulation and displacement, and skeletal maturity. RESULTS: All patients had acceptable alignment at 3-5-day follow-up. Two study patients and one control required re-manipulation at subsequent follow-up (p = 0.34). All other patients in both groups who were seen at follow-up had satisfactory healing and function at 6-8 weeks after injury. Three study patients and 4 controls had an unscheduled outpatient visit to the Emergency Department (ED) for cast-related problems (p = 0.80). None of these patients developed compartment syndrome. The mean length of stay in the ED was lower in the study group than in the control group (3.1 h compared to 5.1 h, respectively; p = 0.0026). The mean facility charge also was lower in the study group ($2182.50 compared to $3031 in the control group; p = 0.0006). CONCLUSIONS: Our results suggest that emergency physicians may be able to successfully provide restorative care for distal forearm fractures using closed reduction technique. Care rendered by emergency physicians was associated with a shorter length of stay and lower facility charges.
Subject(s)
Casts, Surgical , Emergency Service, Hospital , Fractures, Closed/surgery , Radius Fractures/surgery , Adolescent , Child , Child, Preschool , Female , Fluoroscopy , Follow-Up Studies , Humans , Male , Retrospective Studies , Treatment OutcomeABSTRACT
As the size of biomedical implants and wearable devices becomes smaller, the need for methods to deliver power at higher power densities is growing. The most common method to wirelessly deliver power, inductively coupled coils, suffers from poor power density for very small-sized receiving coils. An alternative strategy is to transmit power wirelessly to magnetoelectric (ME) or mechano-magnetoelectric (MME) receivers, which can operate efficiently at much smaller sizes for a given frequency. This work studies the effectiveness of ME and MME transducers as wireless power receivers for biomedical implants of very small (<2 mm³) size. The comparative study clearly demonstrates that under existing safety standards, the ME architecture is able to generate a significantly higher power density than the MME architecture. Analytical models for both types of transducers are developed and validated using centimeter scale devices. The Institute of Electrical and Electronics Engineers (IEEE) and the International Commission on Non-Ionizing Radiation Protection (ICNIRP) standards were applied to the lumped elements models which were then used to optimize device dimensions within a 2 mm³ volume. An optimized ME device can produce 21.3 mW/mm³ and 31.3 ïW/mm³ under the IEEE and ICNIRP standards, respectively, which are extremely attractive for a wide range of biomedical implants and wearable devices.
ABSTRACT
Previous research into pollen content of tobacco resulted in a debate. We address this debate and determine that pollen analysis may be able to assist with identifying geographical origin of tobacco. However, the value of any results should be assessed on a case-by-case regional basis until sufficient database information is available for an objective interpretation to be undertaken on a global basis. As a first step toward developing comparative data for South America, we analyzed a tobacco sample from Brazil in an effort to identify signature taxa from the state of Minas Gerais. We also assessed the role of honey additives to tobacco to assess this issue. Comparing the data to previously published data, we conclude that pollen signatures can distinguish broad geographic areas. We conclude that this forensic interpretation framework needs to be developed in context of the National Academy of Sciences recommendations for tightening methods in forensic science.