Y-STRs in forensic medicine: DNA analysis in semen samples of azoospermic individuals

ABSTRACT: The incidence of rape has increased, especially in metropolitan areas, such as the city of São Paulo. In Brazil, studies about it have shown that the majority of this type of crime is committed by the relatives and persons close to the victim. This has made the crime more difficult to be denounced, as only 10% of the cases are reported to competent police authorities. Usually, cytological exams are carried out in sex crime investigations. The difficulty in showing the presence of spermatozoa is frequent, but it does not exclude the presence of male DNA. The absence of spermatozoa in material collected from rape victims can be due to several factors, including the fact that the agressor suffers from azoospermia. This condition can be the result of a successful vasectomy. As the majority of DNA in the ejaculation sample is from spermatozoa, there is much less DNA to be analyzed. This study presents the application of Y‐STRs (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, and DYS393) in DNA analysis of sperm samples from 105 vasectomized men. The study demonstrated a great variation in DNA concentration. DNA extraction and amplification was possible in all sperm samples even in the absence of spermatozoa. The same profile was observed, for each individual, from DNA extracted from blood, pre‐ and postvasectomy semen samples. The use of markers specific for Y chromosome in sex crime cases, especially in the absence of spermatozoa, is very important, mainly because in most situations there is a small quantity of the agressor's DNA in the medium and a large quantity of the victim's DNA.     

Taphonomic changes to blunt force trauma: a preliminary study

This study examines the effects of taphonomic processes on blunt force trauma (BFT) through an experimental study involving pig heads. Of particular concern is the possibility that taphonomic changes can create pseudo-trauma and/or conceal evidence of actual trauma. BFT was inflicted on 10 pig skulls using a hammer. The skulls were subsequently exposed to the environment for 12 months. Seven taphonomic changes were evaluated: the freeze-thaw cycle; rodent gnawing; carnivore scavenging; presence/weight of soil; presence/weight of rain and snow; movement/displacement of bones; and discoloration due to sun bleaching and grass staining. Taphonomic effects varied between cancellous, compact, fresh, and degreased bone. Freezing and thawing, exposure to rain and snow, movement of the skulls, and soil erosion altered and, in some cases disguised, pre-existing trauma. Rodent and carnivore activity did not obliterate evidence of BFT. Recommendations for evaluating BFT on remains affected by taphonomic processes are presented. As each taphonomic process outlined by this study has the potential to disguise antemortem injury, the authors propose that one must carefully examine large, circular openings in the skull that may represent the remnant evidence of BFT.     

Evaluation of the Suchey-Brooks method for aging skeletons in the Balkans

This study has been carried out to examine whether the Suchey-Brooks (S&B) methods could be successfully applied in age assessment of populations from the Balkans. The known-age sample consists of 33 females and 52 males pairs of pubic bones collected from the autopsy cases. Age estimation by S&B method showed an accuracy of 89.74% in males and 72.0% in females. Statistical analysis showed a positive correlation between the actual age of the investigated individuals and age phases obtained by the S&B method, although the mean values of the sixth age category differed significantly compared with the original model. The most reliable indicators in both sexes were the relief of the symphyseal surface, lipping, symphyseal rim, and dorsal margin. The discriminating power of these indicators was the least reliable in distinguishing S&B phases 2 and 3. Based on these results, the appropriate recommendations for aging Serbian populations are made. There was a good agreement between two observers (kappa=0.726).     

Mole guns in Turkey in 2003-2005

This study describes the frequency of mole guns in Turkey by examining the cases sent to the Council of Forensic Medicine of Turkey between 2003 and 2005. In total, 11 mole guns were examined. Mole guns are manufactured to be used as a trap against detrimental animals. Although they are not meant to be used as a firearm, they are able to cause death. Mole guns appearing in regular casework were evaluated in terms of type of the gun, number of barrels, size and caliber, rifling, design, mechanism, fitness for use, legality, and geographical distribution. Ninety-one percent of the guns were 12 gauge. Most commonly they originate from Inner Anatolia. Mole guns are typically handmade. Some examples of injuries and deaths caused by mole guns are also offered.     

Waxing grave about adipocere: soft tissue change in an aquatic context

When postmortem environmental conditions are "just right," according to the "Goldilocks Phenomenon," soft tissues (and associated fatty acids) are converted into and preserved as adipocere. To better understand this conversion process and the development of adipocere three human cadavers were immersed in outside, water-filled pits for over 3 months to observe adipocere formation in an underwater context simulating actual field conditions. Recordings of environmental conditions showed that temperatures were between 21 degrees C and 45 degrees C, a range sufficient for the growth of Clostridium perfringens. Chemical analysis of liquid and tissue samples revealed an increase in palmitic acid and decrease in oleic acid. This study tracked the remarkable gross morphological changes that can occur in human bodies subjected to an aquatic postmortem environment. The results support the "Goldilocks Phenomenon" and substantiate previous findings that the presence of bacteria and water is crucial for adipocere to form.     

Real-time polymerase chain reaction quantification of canine DNA

The accurate quantification of target DNA is an important step in the short tandem repeat analysis of forensic biological samples. By utilizing quantification data to control the amount of template DNA in the polymerase chain reaction (PCR), forensic scientists can optimize testing and minimize the consumption of limited samples. The ability to identify and quantify target DNA in mixed-species samples is crucial when it may be overwhelmed by nontarget DNA, as in cases of dog attack. We evaluated two quantitative real-time PCR assays for dynamic range, species specificity, and inhibition by humic acid. While both assays proved to be highly sensitive and discriminating, the Melanocortin-1 Receptor (MC1R) gene Taqman assay had the advantages of a shorter run time, greater efficiency, and safer reagents. In its application to forensic casework, the MC1R assay has been advantageous for quantifying dog DNA in a variety of mixed-species samples and facilitating the successful profiling of individual dogs.     

Review of pathologic findings in remnant hearts following valve donation

The failure of medical examiners/coroners (ME/C) to allow heart valve donation is a major problem encountered by tissue agencies. Even though many ME/C favor tissue donation they remain responsible for determination of cause and manner of death. In 2001, the Jesse E. Edwards Registry of Cardiovascular Disease was approached by one of the nation's largest tissue procurement agencies (The American Red Cross--ARC) for the purpose of performing cardiovascular pathologic examinations following valve donation. The affiliation existed from October 2001 to January 2005. This study was undertaken to review all 593 postvalve recovery heart remnants received during that time period to tabulate the abnormalities identified and to determine whether donation interfered with the determination of cause of death. For each case, a preliminary cause of death was provided by the ARC. The decedent's body height and weight were also provided. Using the preliminary cause of death, the 593 cases were divided into natural and nonnatural manner of death groups. This division of the cases resulted in 106 cases placed in the natural manner of death group and 487 cases in the nonnatural manner of death group. For each case, all cardiac findings including significant conditions, additional findings, incidental findings, and congenital abnormalities were tabulated. Within the natural manner of death group, 15 cases had a noncardiac cause of death and 91 cases had a cause of death suspected to be cardiac related. In the 91 cases, a total of 132 significant cardiac findings were identified and there were six structurally normal hearts including two infants. In the nonnatural manner of death group, 214 significant cardiac findings were identified and 222 cases had a structurally normal heart. In both natural and nonnatural groups, the most common cardiac abnormality was atherosclerotic coronary artery disease. Other frequently encountered conditions were also identified including 11 cases with acute angle of origin of a coronary artery (five cases natural group; six cases nonnatural group). An important feature of this review was the recognition of potentially inheritable conditions that were diagnosed in both natural and nonnatural manner of death groups. There were three cases of hypertrophic cardiomyopathy (one natural; two nonnatural), three cases of arrhythmogenic right ventricular cardiomyopathy (one natural; two nonnatural), and one case of mitral valve prolapse (natural). In reviewing these cases, we did not feel that valve donation severely impaired cardiac pathologic examination. The benefits of cardiovascular pathologic examination by a cardiac pathologist include the identification of significant and incidental findings and recognition of potentially inheritable conditions.     

Virtual autopsy and forensic identification-practical application: a report of one case

The body of an unidentified elderly woman was found trapped in a floodgate. Prior to autopsy, full-body multislice computed tomography (MSCT) was performed for study of bone lesions and cause of death. Age was estimated by analysis of the sternal end of the fourth rib and of the pubic symphyseal medial articular surfaces. The results were then compared with the autopsy findings. MSCT was superior to autopsy in diagnosis of traumatic bone lesions and also revealed dental anomalies and signs of drowning. Age estimation gave a similar result for both methods. This case report illustrates the potential value of MSCT for medico-legal investigations of death: diagnosis of injuries, possibility of determining the cause of death, and anthropological study in order to estimate age or to visualize features likely to enable identification of a corpse.     

Age estimation by pulp/tooth ratio in canines by peri-apical X-rays

Estimation of age in individuals has received considerable attention in forensic science, in which it is a widely used method for individual identification, together with paleo-demographic analyses to establish mortality patterns in past populations. The present investigation, which is a continuation of a previously published pilot study, was conducted to examine the possible application of the pulp/tooth area ratio by peri-apical images as an indicator of age at death. A total of 200 peri-apical X-rays of upper and lower canines were assembled from 57 male and 43 female skeletons of Caucasian origin, aged between 20 and 79 years. They belong to the Frassetto osteological collection of Sassari (Sardinia) and are housed in the Museum of Anthropology, Department of Experimental and Evolutionistic Biology, University of Bologna. For each skeleton, dental maturity was evaluated by measuring the pulp/tooth area ratio on upper (x(1)) and lower (x(2)) canines. Very good agreement was found between intraobserver measurements. Statistical analysis was performed in order to obtain multiple regression formulae for dental age calculation, with chronological age as dependent variable, and gender, and upper and lower canines as independent variables. Stepwise regression analysis showed that gender did not contribute significantly to the fit (p=0.881) whereas variables x(1) and x(2) and the first-order interaction between them did. These two variables explained 92.5% of variations in estimated chronological age and the residual standard error was 4.06 years. Lastly, two simple linear regression equations were obtained for age estimation using canines from the maxilla and mandible separately. Both models explained 86% of variations in estimated chronological age and allowed an age-at-death estimate with a residual standard error of about 5.4 years.     

Mitochondrial DNA amplification success rate as a function of hair morphology

This study examines the amplification success rate of mitochondrial DNA from human head hair with respect to their potential for forensic application. Mitochondrial DNA was isolated using a Chelex-based extraction method and amplified using the LINEAR ARRAY duplex PCR system. The particular focus of this study was to characterize the morphological features of human head hair in order to further the understanding of the factors that influence amplification success rate in hair tissue using the LINEAR ARRAY duplex PCR system. 2554 head hairs from 132 individuals representing four population groups were amplified. The hair samples were characterized as follows: 1251 were identified microscopically as telogen hairs and 1303 were classified as hairs without roots (removed before extraction). Amplification success was assessed as a function of several independent variables: morphological characteristics; telogen root versus no root; donor age; scalp origin; use of cosmetic hair treatments; and race of the donor. The results show that a positive correlation exists between amplification success and the presence of a telogen root. Combining the amplification success with either the original or optimized protocol, telogen hairs result in an overall success rate of 77.5% compared with 65% for hairs with no roots. Controlling for telogen hairs, the findings indicate that the overall success rate is independent of cosmetic hair treatments; medulla structure; shaft length, diameter, and volume; and scalp origin. Conversely, the age of the donor, the race of the donor, and hair pigmentation all contribute to a variation in amplification success rate.