DNA extraction: an anthropologic aspect of bone remains from sixth- to seventh-century ad bone remains

In the archeological site of the early Christian Episcopal complex of Saint Peter, in Canosa di Puglia (Bari, Italy), during the operations of archaeological excavations, tombs were discovered. They were dated between the sixth and seventh centuries ad with carbon 14 methodology. Five skeletons were found in the 5 tombs: 28A: male individual, 43 years old. The height was 170 cm; the biomass was 65.7 kg. The analysis of the bones indicated several noteworthy pathologies, such as a number of hypoplasia lines of the enamel, the presence of Schmorl hernias on the first 2 lumbar vertebrae, and the outcome of subacromial impingement syndrome. 28E was a male individual, with a biologic age of death of between 44 and 60 years. The height was 177 cm. He had a posttraumatic fracture callus of the medial third of the clavicle, with an oblique fracture rima. 29B was a female individual, 44-49 years old. The height was 158.8 cm; the biomass was 64.8 kg. There was Wells bursitis on the ischial tuberosity on both sides. 29E was a male individual, 45-50 years old. The height was 169.47 cm; the biomass was 70.8 kg. The third and the fourth vertebrae showed Baastrup syndrome (compression of the vertebral spine). There were radiologic signs of deformity on the higher edge of the acetabula and results of frequent sprains of the ankles. 31A was a male individual, 47-54 years old. The height was 178.65 cm; the biomass was 81 kg. The vertebral index showed a heavy overloading in the thoracic lumbar region. There were bony formations under the periosteum on both on the higher and medium facets of the first metatarsus and on the higher and lateral facets of the fifth metatarsus on both sides. As the topography indicates, these small ossifications coincided with the contact points between the back of the foot and parts of the upper shoe. From the osseous remains, in particular from the teeth (central incisors), the DNA was extracted and typed to identify potential family ties among all the subjects. The extraction technique used came from the DNA Promega technique, partially modified by the authors. Stay times of the sample in the extraction buffer were increased and were increased the polymerase chain reaction (PCR) cycles.     

Forensic medical expertise on burned bone remains

The series of 1138 cadaveric humerus and femurs of people of both sexes who died at the age of 17-91 years and 468 animal bones (cow, pig, sheep) was studied. Bones were burned (experimentally) at various temperature values and in different conditions. Investigations were carried out using osteometric, microscopic, microroentgenographic and mathematical methods. Species identification as well as sex and age determination using burned bone remains are possible regardless of the level of their burning. Regression equations to calculate victim's stature according to fragments of burned bones were received.     

Microscopical study on estimation of time since death in skeletal remains

For the purpose of estimating time since death in skeletal remains, postmortem changes in human compact bones were examined by microradiography and electron microscopy. The UV-fluorescence of the peripheral zone of compact bone was also examined by microscopic spectrophotometry. Microradiographic examination revealed no morphological changes in bones left in the open air for long periods, except one of 15 years since death. In bones left in the soil, vacuoles of 5-10 microns diameter, which contained a honeycomb-like structure formed by small vacuoles of 0.5-1 microns diameter, were found in the peripheral zone of the substantia compacta approximately 5 years since death, and in bones of 6 years or more, this change extended to the mid-zone. In bones left in the sea for 4-5 years, vacuoles of 5-10 microns diameter were observed in the outer peripheral zone of the substantia compacta. The relative intensity of UV-fluorescence in bones dwindled with time since death and the correlation coefficient was considerably high.     

Role of orthopedic implants and bone morphology in the identification of human remains

When conventional methods of identification, such as visual recognition and dental comparison, cannot be used to identify a deceased person, it becomes necessary to consider alternative methods. The presence of an orthopedic implant in a body may assist identification if ante-mortem medical records are available for comparison. Another method of identification involves comparison of ante-mortem and postmortem radiographs. Eight cases are reported from Forensic Science SA where the presence of orthopedic implants and/or ante-mortem radiographs were used to try to establish identification. In six cases, positive identification was established, and in two cases with upper limb orthopedic implants, the bones remained unidentified. Manufacturers were unable to provide any information about the distribution and use of the implants that could be of use with identification, as there are no requirements in Australia for individual medical implants to be tracked. Such a system has the potential to aid postmortem identification if serial codes were etched onto implants that could then be traced to manufacturers, surgeons, and recipients of these devices.     

Frequency of bone lesions: an inadequate criterion for gunshot wound diagnosis in skeletal remains

A retrospective study was carried out on 130 fatalities due to gunshot wounds to determine whether the combined absence of bone damage and projectile in a skeleton is sufficient to eliminate a diagnosis of gunshot wound. Our findings showed that bone lesions were present in about 90% of the cases and were associated with intracorporeal projectile(s) in about 70% of the cases. The presence or absence of bone lesions seemed independent of the gun characteristics, the shot conditions, and the type of death. The cause of death was predominantly brain injury in cases with bone lesions whereas thoracic, abdominal, and peripheral vascular causes were more frequently encountered in cases without bone damage. We concluded that the combined absence of bone lesions and intracorporeal projectile (about 5% in our series) cannot exclude a diagnosis of death secondary to gunshot wounds.     

The application of differential thermal analysis and thermogravimetric analysis to dating bone remains.

Thirty-four samples of bone remains of known time since death were studied by using DTA, TGA, and derivative-TGA techniques. The DTA patterns enable us to distinguish recent from old (more than 100 years) bones. The TGA curve is also significant for an extreme series. Both DTA and TGA curves show a correlation that allows us to obtain patterns with high significance for the extreme series. They also make evident the decomposition grade that bone organic material undergoes during the postmortem putrefactive process.     

Typing of deoxyribonucleic acid (DNA) extracted from compact bone from human remains.

The application of deoxyribonucleic acid (DNA) typing methods for the potential identification of unknown human remains was investigated. DNA was isolated from compact bone tissue from badly decomposed bodies and from known and unknown human remains, using a decalcification and ion wash procedure. Restriction fragment length polymorphism (RFLP) analysis of variable number of tandem repeats (VNTR) loci yielded results in some cases, but more often the DNA was too degraded to produce RFLP patterns. No RFLP profiles could be obtained from putrefied soft tissues. However, DNA extracted from compact bone tissue of human remains up to eleven years old was successfully amplified using the polymerase chain reaction (PCR) for the VNTR loci D1S80, D17S5, COL2A1, and APO B, as well as the HLA-DQ alpha locus. This is especially significant, since PCR results were obtained from those samples whose DNA had been degraded substantially and had yielded no RFLP patterns. All DNA types determined from the compact bone tissue from decomposed bodies whose identification had been established first by other means (and whose parents or offspring were available for typing) demonstrated mendelian inheritance of the alleles of the loci analyzed. These results suggest that amplification and typing of DNA extracted from compact bone of human remains could be useful in establishing the identity of a person, as well as in excluding possible false identifications.     

Evidence of arteriosclerosis in cremated remains.

Severely burned or cremated human remains (i.e., cremains) are among the most challenging cases investigated by forensic pathologists, odontologists, and anthropologists. Exposure to extreme heat of long duration destroys all organic components of the body, leaving only the inorganic component of the skeleton. The numerous calcined osseous or dental fragments that remain after the cremation process rarely convey useful information to the investigator. In most cases, it is associated nonosseous artifacts that provide evidence of the decedent's identity. In a case investigated at the University of Florida's C. A. Pound Human Identification Laboratory, as well as in several cases examined during data collection for a research project, several fragments were identified as calcined plaque tubules from sclerotic blood vessels. These tubules provide direct evidence that the cremated individual had a variety of arteriosclerosis.     

Cocaine in decomposed human remains.

From March 1988 through March 1990, at the Philadelphia Medical Examiner's Office toxicology laboratory, samples from 77 decomposed human bodies were tested for the presence of cocaine, employing gas chromatography/mass spectrometry (GC-MS). The material analyzed included decomposed soft tissue, bloody decomposition fluid, mummified tissue, maggots, and beetle feces. Twenty-two cases (28.6%) were positive for cocaine, many of these cases in states of advanced decomposition. These findings indicate the usefulness of testing decomposed tissue for cocaine in all cases where its presence is suspected. This is contrary to what might be expected, since cocaine is generally labile and rapidly broken down by both enzymatic and nonenzymatic mechanisms.     

Heterotopic ossification in unidentified skeletal remains

Heterotopic ossification is a benign, ectopic bone growth that develops in muscle and other soft tissue. The exact cause is poorly understood, but it is a rarely serious complication of soft tissue trauma. Its most common form, myositis ossifications traumatica, occurs as a secondary complication of direct muscle injury. However, other forms are less common and can result from specific pathologic conditions, such as spinal cord trauma and metabolic disorders. In patients who have had spinal cord injury and subsequent paraplegia, heterotopic ossification often results in ankylosis of the hip and a loss in range of motion. Ectopic ossification occurs below the injury site, and, although the specific muscle groups can vary, it usually involves those for which the origin and insertion involve the anterior pelvis and proximal femur. In dried bone, heterotopic ossification can appear as a smooth, irregularly shaped benign tumor of mature bone, extending from the surface but not invading the cortical bone. These tumors range in size from a few millimeters to several centimeters. Because heterotopic ossification is often associated with specific types of injuries, it has a unique anthropological use in forensic cases.     

Physical and mechanical degradation of shirting fabrics in burial conditions

The current focal areas within forensic textile science are fibre identification and assessment of the method of damage to fabrics. This paper investigates fabric degradation within clandestine burials. The fabrics considered in this paper, unlike previous archaeological studies, are a modern polyester-cotton blend (65%/35%) and a 100% cotton fabric both of which are commonly used for men's shirting fabrics in the UK. Three laundering conditions were investigated (i) not-laundered, (ii) laundered 6 times, and (iii) laundered 60 times; this represented varying conditions of fabric upon clothing deposition. The two burial conditions; sand and clay, were selected as extremes of soil type. The deposition times (15 and 30 days) were based on a study of clandestine burials in UK crimes. There were clear differences in how polyester-cotton and cotton stained within the two different soil conditions, polyester-cotton becoming extensively stained after a 30-day deposition in sand. The tear force required to tear the fabric after deposition, suggested that polyester/cotton fabrics were consistently weaker after burial, regardless of soil type and deposition period. There was also significant damage caused to not-laundered cotton fabrics after a 30-day deposition in clay. This work indicates that common apparel fabrics can degrade in relatively short times when buried.