Body fluid identification by integrated analysis of DNA methylation and body fluid-specific microbial DNA

Identification of body fluids found at crime scenes provides important information that can support a link between sample donors and actual criminal acts. Previous studies have reported that DNA methylation analysis at several tissue-specific differentially methylated regions (tDMRs) enables successful identification of semen, and the detection of certain bacterial DNA can allow for identification of saliva and vaginal fluid. In the present study, a method for detecting bacterial DNA was integrated into a previously reported multiplex methylation-sensitive restriction enzyme-polymerase chain reaction. The developed multiplex PCR was modified by the addition of a new semen-specific marker and by including amplicons for the 16S ribosomal RNA gene of saliva- and vaginal fluid-specific bacteria to improve the efficacy to detect a specific type of body fluid. Using the developed multiplex system, semen was distinguishable by unmethylation at the USP49, DACT1, and PFN3 tDMRs and by hypermethylation at L81528, and saliva could be identified by detection of saliva-specific bacteria, Veillonella atypica and/or Streptococcus salivarius. Additionally, vaginal fluid and menstrual blood were differentiated from other body fluids by hypomethylation at the PFN3 tDMR and the presence of vaginal fluid-specific bacteria, Lactobacillus crispatus and/or Lactobacillus gasseri. Because the developed multiplex system uses the same biological source of DNA for individual identification profiling and simultaneously analyses various types of body fluid in one PCR reaction, this method will facilitate more efficient body fluid identification in forensic casework.     

Evaluation of commercial multi-drug oral fluid devices to identify 39 new amphetamine-designer drugs

Recently, the diffusion on the black market of new psychoactive substances not controlled and often sold as ‘legal highs’, is exponentially increasing in Europe. Generally, the first analysis for these drugs involves an immunoassay screening in urine or plasma. Actually, there is growing interest in the use of oral fluid (OF) as alternative specimen over conventional biological fluids for drug testing, because of the significant advantages, as a non-invasive collection under direct observation without undue embarrassment or invasion of privacy, and a good correlation with plasma analytical data. Few assays have been developed for detection of new psychoactive compounds in biological samples, so it is important to investigate how they may or may not react in pre-existing commercial immunoassays. In this paper, two different multi-drugs oral fluid screen devices (OFDs) (Screen® Multi-Drug OFD and GIMA One Step Multi-Line Screen Test OFD) were evaluated to determine the cross-reactivity of thirty-nine new amphetamine designer drugs, including twelve substances officially recognized as illicit by italian legislation. Cross-reactivity towards most drugs analyzed was <1 in assays targeting amphetamine (AMP) or methamphetamine (MET). Only two (p-methoxyamphetamine and p-methoxymethamphetamine) of all tested amphetamines gave a positive result.     

Using edge enhancement to identify subtle findings on soft-copy neonatal chest radiographs.

OBJECTIVE: The purpose of this study was to evaluate whether edge enhancement could improve the visibility of subtle findings on soft copies of neonatal chest radiographs. MATERIALS AND METHODS: Two radiologists reviewed 82 soft-copy neonatal chest radiographs before and after the application of edge enhancement on our picture archiving and communication system (PACS). The visibility of a pneumothorax (n = 22), central venous catheter (n = 32), umbilical arterial catheter (n = 36), endotracheal tube (n = 40), and normal anatomic structures (the minor fissure, anterior segmental bronchus of the right upper lobe, and aortic arch, n = 57) was evaluated. Six of 22 soft-copy images depicting a pneumothorax were excluded from the evaluation of image quality either because of the large size of the pneumothorax itself (n = 7) or because of the lack of confirmatory evidence that would have been provided by an additional lateral decubitus (n = 6) or cross-table lateral radiograph (n = 3). Image quality was evaluated by visual grading analysis. RESULTS: The visibility of a pneumothorax (p < 0.01), vascular catheters (p < 0.001), the minor fissure (p < 0.001), and the anterior segmental bronchus of the right upper lobe (p < 0.001) improved significantly after applying edge enhancement to soft copies of neonatal chest radiographs, whereas the visibility of the aortic arch did not improve. Evaluations of the improvements in the visibility of the endotracheal tube were inconsistent. CONCLUSION: Application of edge enhancement to soft copies of neonatal chest radiographs helps radiologists to identify small pneumothoraces, vascular catheters, and delicate normal structures, thereby improving the detection of subtle chest findings in the neonatal intensive care unit.     

16S核糖体DNA Amplicon测序法分析2型糖尿病患者口腔微生物多样性

目的探讨2型糖尿病患者口腔微生物多样性及结构的差异。方法采集23例2型糖尿病患者与23名正常人的唾液与龈上菌斑样本,提取口腔菌群的总DNA,利用16S核糖体DNA（16S ribosome DNA,16SrDNA）Amplicon测序法进行测序,使用Pandaseq、Qiime等软件分析菌群多样性及结构。结果2型糖尿病患者和正常人口腔菌群丰度及优势菌属差异无统计学意义(P>0.05)。2型糖尿病患者样本中布雷德菌属、瘤胃菌科、幽门螺杆菌科丰度较高,正常人样本中韦荣菌科、Selenomonadales目、Negativicutes纲、月形单胞菌属、丁酸弧菌属、Anaeroglobus属、Candidatus Saccharibacteria门、Saccharibacteria_genera_incertae_sedis属、诺卡菌科、气微菌属丰度较高,差异有统计学意义（P<0.05）。结论用16SrDNA Amplicon测序法分析2型糖尿病患者与正常人口腔菌群结构具有相似性,幽门螺杆菌与2型糖尿病之间的关系还有待于进一步的研究。     

Incidental findings in the use of DNA to identify human remains: An ethical assessment

DNA analysis is increasingly used to identify the remains of victims of conflicts and disasters. This is especially true in cases where remains are badly damaged and fragmented, or where antemortem records are unavailable. Incidental findings (IFs)—that is, genetics-related information for which investigators were not looking—may result from these identification efforts employing DNA analysis. Because of the critical role played by family members of the missing in identification efforts, as well as the familial nature of DNA, identification initiatives employing DNA analysis are particularly prone to reveal IFs about familial relationships, such as misattributed paternity or false beliefs about sibling relationships. Despite forensic scientists’ widespread awareness of the possibility of generating IFs, to date there has been relatively little explicit guidance about their management. This paper fills that gap. It offers substantive guidance about the ethical management of IFs in this context. To ensure that the analysis addresses actual needs and practices in the field, one author (JDA) conducted semi-structured interviews with key informants from six regionally diverse organizations involved in post-conflict or post-disaster identification efforts. The paper first describes how methods of DNA analysis give rise to IFs. Next, it explains the importance of developing an ethically justified general policy for managing IFs and discusses features of DNA identification efforts that are relevant to such a policy. Then it presents an argument in support of a general policy of nondisclosure—specifically, that considerations of fair access to the individual and social benefits of identification efforts, and the concern to minimize and fairly distribute the risks of participation, support a policy of nondisclosure. It concludes by considering some implications of this argument for the choice among scientific practices involved in using DNA analysis to identify human remains, as well as for managing non-genetic incidental findings.     

Using pullback pressure measurements to identify venous stenoses persisting after successful angioplasty in failing hemodialysis grafts

OBJECTIVE: We used pullback pressure measurements to identify venous stenoses persisting after angioplasty of failing hemodialysis grafts. MATERIALS AND METHODS: Fifty angioplasty procedures were performed in 32 patients with elevated venous pressures at dialysis. Grafts were initially evaluated on digital subtraction angiography, and all stenoses measuring greater than 50% on angiography underwent angioplasty. In successful cases (residual stenosis < 30%), pullback pressure measurements were obtained from the superior vena cava to the graft to identify hemodynamically significant (> 10 mm Hg) stenoses. These lesions were then treated with repeated angioplasty. RESULTS: Hemodynamically significant stenoses with a gradient range of 10-27 mm Hg (mean, 16 mm Hg) were found in nine (18%) of 50 procedures. All gradients occurred at sites of previous angioplasty. Repeated angioplasty of these stenoses performed with larger angioplasty balloons reduced gradients to less than 3 mm Hg in six stenoses and to 5 mm Hg in three stenoses. In this subgroup, primary patency was eight (89%) of nine stenoses at 1 month and 2 months and five (56%) of nine stenoses at 6 months. Using life table analysis, we found that primary patency of the entire population was 84% at 1 month, 66% at 2 months, and 47% at 6 months. The mean time between interventions was 6 months, and the thrombosis rate was 0.32 per year. CONCLUSION: Pullback pressure measurements are a useful adjunct to angiography to evaluate the hemodynamic results of angioplasty in patients with failing hemodialysis grafts.     

Using multimedia to support jury understanding of DNA profiling evidence

Jury difficulties in understanding and according suitable weight to probabilistic scientific expert evidence about DNA profiling are well established. Traditional legal aids, including note-taking and allowing jury questions, have had limited success in diminishing their problems. This study examined the potential of multimedia instruction to enhance jury comprehension. Mock jurors reviewed a homicide case in which DNA evidence linked the suspect to the crime scene. Instructional media on ‘Forensic DNA Technology’ and/or ‘Random Match Probability’ accompanied the oral expert evidence to support juror understanding. Pre-test–post-test comparisons of juror conceptions about forensic uses of DNA determined whether these multimedia facilitated juror comprehension. Analyses of responses to the expert evidence acknowledged the incidence of ‘CSI’ effects in which jurors have increased expectations of scientific evidence after viewing popular crime investigations on television. Since multimedia instruction for juries can apply to all forms of forensic evidence, we propose the use of structured instructional media for courts developing policies on uses of visual evidence.     

Using breakeven analysis to examine price sensitivity

Increased competition and managed care have brought about a need to reexamine how operating expenses are portrayed. Distinguishing between fixed and variable costs and using breakeven analysis can determine the implications of price concessions that may be requested by insurance companies. This technique also identifies areas of operating that hurt profitability of a specific imaging site or a modality within a single site. Breakeven analysis can also be used to investigate the effect of capitation and to strategize in an environment where commercial insurers sometimes establish fees that are so low, providers would prefer capitated contracts. In that situation the only option may be to manage utilization to enhance profit. This article includes financial models that can be used to simulate the implications of changes in the average income per scan (Table 1) and to examine the profitability of each segment of service within an imaging center (Table 2). Table 2 allows management to trendline each modality and examine cost chracteristics. That enables those responsible for overseeing operations to isolate and understand problem areas.     

Forensic hair analysis to identify animal species on a case of pet animal abuse

As part of an investigation of a case of pet animal abuse, we attempted to identify small mammalian species by morphological analysis and single-nucleotide polymorphism (SNP) typing of the cytochrome b gene using guard hairs as an analytical material. Guard hair samples from several species were measured for length, width, medulla formation, and cuticle scale pattern under a light microscope or scanning electron microscope. These samples were also analyzed for SNPs in the cytochrome b gene using a multiplex single-base primer extension reaction. Morphological analysis of cuticle scale pattern and medulla formation was able to discriminate ferret hairs from other hair samples that included rabbit, gerbil, degu, and Djungarian hamster. However, this also revealed a similarity of the guard hairs of the ferret (Mustela putorius furo) and Japanese weasel (Mustela itatsi). Although at three sites, the nucleotide color signals of SNPs in the cytochrome b gene could be used to discriminate completely among human, dog, and gerbil, the signals for cat, ferret, and Japanese weasel occurred at the same nucleotide sites. Unfortunately, no signals were obtained from degu, Djungarian hamster, and rabbit hairs. Although the discriminated hair samples were 100% identical to those of the ferret, there was only a 5% difference from Japanese weasel in the partial sequence of the cytochrome b gene. Construction of a database of mammalian hairs would be useful not only in forensic science, but also for investigating smuggling of endangered species in contravention of the Washington Convention.     

Role of oral fluids in DNA investigations

The assay of oral fluid (OF), a biofluid historically well-studied biochemically and physiologically, is a growing area of research with implications for basic and clinical purposes. In the last decade, it has gained considerable attention and lately, the use of OF has provided a substantial addition as an investigative tool in forensic and/or legal procedures. This article is an appraisal of various applications of OF sourced DNA in the field of forensic analysis. We have discussed the significance of different collection methods and their variations along with the application of specific analytical methods based on the condition of the sample. It is likely that the germaneness of OF assays will continue to expand thus providing a new instrument for investigation in criminal/legal proceedings.     

航海人员洁治术对干槽症的影响及原因分析

目的　探讨干槽症发病的相关因素和预防航海人员干槽症发生的方法。方法　对 4 6 8例需要拔除双侧下颌阻生智齿的航海人员在无拔牙禁忌证的情况下分次进行拔牙手术 (相隔时间 2个月以上 )。第 1次拔牙前不进行特殊术前准备 ,第 2次术前进行全口龈上下洁治术 ,术后 3～ 5 d拔牙。 2次拔牙前均牙菌斑取样检测幽门螺杆菌 (Helicobacter pylori,H.Pylori)。结果　第 1次拔牙后 78颗发生干槽症 ,占 16 .6 7% ;第 2次拔牙后仅 9颗发生干槽症 ,占 1.92 % ,差异非常显著 (P<0 .0 1)。牙菌斑 H.py-lori检测结果 2次拔牙前阳性率分别为 5 4 .2 7% (2 5 4 / 4 6 8)和 14 .74 % (6 9/ 4 6 8) ,差异亦有显著性 (P<0 .0 5 ) ,且所有发生干槽症的手术拔牙前牙菌斑 H.Pylori检测均为阳性。结论　口腔幽门螺杆菌感染可能是航海人员干槽症发病率高的原因 ,术前口腔洁治可以有效预防拔牙术后干槽症的发生。     

Cluster analysis using physical performance and self-report measures to identify shoulder injury in overhead female athletes

Objectives

To evaluate the diagnostic validity of the Kerlan-Jobe orthopedic clinic shoulder and elbow score (KJOC) and the Closed kinetic upper extremity stability test (CKCUEST) to assess functional impairments associated with shoulder injury in overhead female athletic populations.

应用定量CT纹理分析鉴别肾上腺肿瘤

目的 探讨应用CT纹理分析鉴别肾上腺乏脂腺瘤、嗜铬细胞瘤与肾上腺转移瘤的可行性.方法 回顾性分析66例肾上腺乏脂腺瘤、98例嗜铬细胞瘤和101例肾上腺转移瘤CT图像.所有患者均行腹部CT平扫与肾上腺增强扫描.应用TexRAD软件分析CT图像的纹理特征并比较3种肾上腺病变之间各纹理参数间的差异.结果 平扫CT图像上,3种病变的平均灰度值(Mean)和峰度(Kurtosis)在所有过滤值(SSF)纹理图像上(SSF 0~6)均有显著差异(P<0.05),标准差(SD)在精细和粗糙纹理上(SSF 2、6)差异有统计学意义(P<0.05).肾上腺乏脂腺瘤和转移瘤的熵(Entropy)(SSF 0~3,5~6)和正像素的平均值(MPP)(SSF 0~2,4~6)均明显低于嗜铬细胞瘤(P<0.05).3种病变的偏度(Skewness)(SSF 0~3)有显著统计学意义(P<0.05),嗜铬细胞瘤最低而转移瘤最高.增强CT图像上,3种病变的Mean、SD、Entropy和MPP在所有等级纹理图像上(SSF 0~6)均有显著统计学意义(P<0.05),4种纹理参数均在嗜铬细胞瘤上最高、在转移瘤上最低.肾上腺转移瘤的Skewness(SSF 0)和Kurtosis(SSF 0, 2)明显低于乏脂腺瘤和嗜铬细胞瘤(P<0.05).结论 肾上腺乏脂腺瘤、嗜铬细胞瘤与肾上腺转移瘤的CT纹理分析参数有显著性差异,CT纹理分析具有鉴别以上3种病变的潜在临床应用价值.