Estimating the Temperature of Heat‐exposed Bone via Machine Learning Analysis of SCI Color Values: A Pilot Study

Determining maximum heating temperatures of burnt bones is a long‐standing problem in forensic science and archaeology. In this pilot study, controlled experiments were used to heat 14 fleshed and defleshed pig vertebrae (wet bones) and archaeological human vertebrae (dry bones) to temperatures of 400, 600, 800, and 1000°C. Specular component included (SCI) color values were recorded from the bone surfaces with a Konica‐Minolta cm‐2600d spectrophotometer. These color values were regressed onto heating temperature, using both a traditional linear model and the k‐nearest neighbor (k‐NN) machine‐learning algorithm. Mean absolute errors (MAE) were computed for 1000 rounds of temperature prediction. With the k‐NN approach, the median MAE prediction errors were 41.6°C for the entire sample, and 20.9°C for the subsample of wet bones. These results indicate that spectrophotometric color measurements combined with machine learning methods can be a viable tool for estimating bone heating temperature.     

The Use of Near‐Infrared Photography to Image Fired Bullets and Cartridge Cases

An imaging technique that is capable of reducing glare, reflection, and shadows can greatly assist the process of toolmarks comparison. In this work, a camera with near‐infrared (near‐IR) photographic capabilities was fitted with an IR filter, mounted to a stereomicroscope, and used to capture images of toolmarks on fired bullets and cartridge cases. Fluorescent, white light‐emitting diode (LED), and halogen light sources were compared for use with the camera. Test‐fired bullets and cartridge cases from different makes and models of firearms were photographed under either near‐IR or visible light. With visual comparisons, near‐IR images and visible light images were comparable. The use of near‐IR photography did not reveal more details and could not effectively eliminate reflections and glare associated with visible light photography. Near‐IR photography showed little advantages in manual examination of fired evidence when it was compared with visible light (regular) photography.     

Storage of Second World War bone samples: Bone fragments versus bone powder

Bone samples may yield low-quality and low-quantity DNA and duplicated analyses of different genetic markers have to be performed for identification of missing persons. Mostly no DNA extract is left after analyses and efficient storage of bones is needed to ensure the stability of the sample over time for retesting using new markers and new technologies. Usually not all of the bone powder prepared in grinder is used for extraction and rest can be stored for future analyses. After molecular genetic analyses of 88 victims of Second World War (WWII) Konfin I mass grave in Slovenia (performed in 2009), fragments of femurs and bone powder that were left were stored at -20 °C. Some authors reported that long-term storage of powder results in the reduction of DNA preservation and its degradation (even at low temperature), explained by an increase in oxidative damage as a result of the enormous increase in exposed surface area. Consequently, grinding of bones as shortly prior to DNA extraction was recommended. The goal of our study was to explore the difference in DNA yield between bone fragment and bone powder frozen for 10 years. 57 WWII femurs were examined and DNA extracted from each of them using bone fragment (piece sampled next to the one used in 2009) and bone powder obtained in 2009, both taken out of freezer after 10 years of storage. Half gram of bone powder was decalcified using full demineralization extraction method. The DNA was purified in a Biorobot EZ1 (Qiagen) and quantified with PowerQuant kit (Promega). Statistical analysis showed significant difference at the 0.05 level in DNA yield comparing fragments of bones and bone powder stored at -20 °C for 10 years. The results show there is more DNA stored in the bone powder than in the bone fragments. Because of time - consuming powdering procedure we recommend to store not only the fragment of the bone, but obtained bone powder as well.     

The ordinal localization of mono-type foot bones

The osteologic collection of the chair for anthropology, Moscow State University, comprising male (n = 70) and female (n = 14) foot bones from persons aged more than 18-20, was investigated. An appropriate Martin program was fixed to the instep bones (IB) of the proximal and middle toe phalanxes (4 sizes for each bone) with 65 signs being studied in each case. The mathematical data processing was SPSS-aided. Diagnostic models for defining the order IB localization in the proximal and middle phalanxes were elaborated on the basis of discriminative analysis. The classification accuracy is maximal for IB (80-100%) and minimal for the middle phalanxes (26.19-77.38%). The definition of the order localization of phalanxes in toes 2-5 is virtually impossible. The designed method can be used alongside with the routine anatomic-and-morphological findings. The congruence of surfaces in the instep-phalanx and interphalangeal joints must be necessarily checked.     

Characteristics and Frequency of Chipping Effects in Near‐Contact Gunshot Wounds

The presence of “chipping” or “flaking” around the edges of gunshot entry wounds has been described among the characteristics of gunshot wounds in bone. In this study, the real frequency of such a peculiar feature was investigated. The presence of “chipping” was assessed on 22 gunshot wounds fired at a near‐contact range on bovine ribs with 9‐mm bullets. As controls, five samples were shot with a 3 cm range, and five from 40 cm. In 77% of cases shot at near‐contact range, a detachment of small fragments of the upper layers of bone was detected, mainly with a circumferential disposition, whereas this feature was lacking in control samples. The study demonstrated the frequency of “chipping” and that it may probably be due to a combined ballistic effect of impact of the bullet itself and expansion of gases. It may be thus considered indicative of close‐range shots.     

Histological Determination of the Human Origin of Bone Fragments

Abstract:  A frequently encountered task in the forensic scenario is verification of the human origin of severely degraded fragments of bone. In these cases histological methods which consider osteon size and morphology can prove to be useful. The authors in the present study verify the applicability of published algorithms to flat and subadult bones from human, dog, cat, cow, rabbit, sheep, pig, chicken, quail, and turkey samples. Metric analysis was performed on 2031 Haversian canals. Analyses carried out on human samples confirmed a success rate of around 70% on long adult bones; however the percentage of wrong answers was particularly high in the case of newborns and older subadults as well as on flat bones in general. Results therefore suggest that such regression equations should be limited only to bone fragments from long adult bones.     

Reconstructing the Sequence of Events Surrounding Body Disposition Based on Color Staining of Bone*

Abstract: Literature regarding bone color is limited to determining location of primary and secondary dispositions. This research is the first to use bone color to interpret the sequence of events surrounding body disposition. Two scenarios were compared—bones buried and then exposed on the ground surface and bones exposed then buried. Forty juvenile pig humeri with minimal tissue were used in each scenario with an additional 20 controls to determine if decomposing tissue affects bone color. Munsell Color Charts were used to record bone color of surface and 2.5 cm cross‐sections. Results reveal five main surface colors attributed to soil, sun, hemolysis, decomposition, and fungi. Fungi on buried bones suggests prior surface exposure. Cross‐sections of strictly buried bones are identical to buried then exposed bone, stressing the importance of bone surface analysis. Cross‐sectioning may help verify remains have been exposed then buried. Decomposition of excess tissue creates minimal color staining.     

A Novel Method for the Photographic Recovery of Fingermark Impressions from Ammunition Cases Using Digital Imaging

The photographic preservation of fingermark impression evidence found on ammunition cases remains problematic due to the cylindrical shape of the deposition substrate preventing complete capture of the impression in a single image. A novel method was developed for the photographic recovery of fingermarks from curved surfaces using digital imaging. The process involves the digital construction of a complete impression image made from several different images captured from multiple camera perspectives. Fingermark impressions deposited onto 9‐mm and 0.22‐caliber brass cartridge cases and a plastic 12‐gauge shotgun shell were tested using various image parameters, including digital stitching method, number of images per 360° rotation of shell, image cropping, and overlap. The results suggest that this method may be successfully used to recover fingermark impression evidence from the surfaces of ammunition cases or other similar cylindrical surfaces.     

Quantitative Analysis of the Morphological Changes of the Pubic Symphyseal Face and the Auricular Surface and Implications for Age at Death Estimation

Age estimation methods are often based on the age‐related morphological changes of the auricular surface and the pubic bone. In this study, a mathematical approach to quantify these changes has been tested analyzing the curvature variation on 3D models from CT and laser scans. The sample consisted of the 24 Suchey–Brooks (SB) pubic bone casts, 19 auricular surfaces from the Buckberry and Chamberlain (BC) “recording kit” and 98 pelvic bones from the Terry Collection (Smithsonian Institution). Strong and moderate correlations between phases and curvature were found in SB casts (ρ 0.60–0.93) and BC “recording kit” (ρ 0.47–0.75), moderate and weak correlations in the Terry Collection bones (pubic bones: ρ 0.29–0.51, auricular surfaces: ρ 0.33–0.50) but associated with large individual variability and overlap of curvature values between adjacent decades. The new procedure, requiring no expert judgment from the operator, achieved similar correlations that can be found in the classic methods.     

Radiological, forensic, and anthropological studies of a concrete block containing bones

Multidisciplinary forensic, anthropological, and radiological studies of bone fragments encased in a concrete block were carried out to determine whether or not the bones were human. Multislice computed tomography (MSCT) investigation was performed before the bones were removed from the concrete. MSCT study pinpointed the location of the bone fragments within the concrete block, which was helpful for their extraction and recovery, and identified most of their types and nature. Osteological study on dry bones provided more accurate identification of the bones and of their side. According to both methods, the human skeletal remains were compatible with those of a child, aged 8-13 years old, with a minimum height of 128 cm. Neither investigation identified sex or racial phenotype. Both studies identified the skeletal remains as consisting of two animal and five human bones. Furthermore, both methods revealed that the concrete completely encased bones, suggesting a secondary burial.     

Forensic-medical significance of the characteristics of gunshot perforating fractures of the skull bones

The study was made of gunshot perforating fractures of calvarial bones in cases of lethal perforating head wounds in long distance shots from a 9-mm Makarov gun. Morphological characteristics of inlet and outlet perforating fractures and bone fragments were found to differ. Gunshot fractures in calvarial bones inflicted by bullets from a 9-mm Makarov gun were studied in experiments with different kinetic energy. In low kinetic energy, bullets make oval holes on the outer table of the skull and round holes on the inner table with radial prominences on the walls of bullet channels, large bone fragments; in moderate kinetic energy--make round holes in the outer table and oval on the inner table with skew prominences on the walls of the outlet part of the channels, large and middle-sized flat bone fragments; in high kinetic energy injuries on both tables were round, prominences were cross, bone fragments were flat, small and middle-sized.     

Mechano- and morphogenesis of gunshot fractures in the flat bones

Gunshot fractures in flat bones, bullet defects in the continuum were studied in experiment. The laws of gunshot fracture formation were formulated. In far distance, bullet has a direct and radial blow action. The direct blow induces comminution and disintegration of the bone tissue with dislocation of bone fragments; radial blow potentiates bone destruction. Morphological characteristics of gunshot fractures of flat bones depend on the force of the direct and radial impact action and can be used for estimation of the far shot distance, kinetic energy and shape of the head of the shell.     

舌骨的法医人类学研究

目的建立利用舌骨形态测量值判别性别的方法,并探讨其在法医学应用中的价值。方法对于河北地区收集的108例舌骨样本(男78例,女30例)进行观察研究,确立11个观测指标并进行测量,测量值采用SPSS统计软件进行统计学分析,建立利用舌骨形态测量值判别性别的方程,并进行Fisher判别分析。结果在11项观察指标中,8项测量指标男女性别间存在显著性统计学差异(P<0.01);利用8项测量指标中的任一指标或之中的多个指标组合建立线性性别判别方程,方程判别性别的准确率在66%～92%,其中多指标的方程判别性别准确率较单一指标的方程高,最高达到92%。结论舌骨形态存在着性别差异,利用多个舌骨形态学测量值建立的性别判别方程准确率较高,可用于无名尸骨的性别判别。     

Assessment of the Effects Exerted by Acid and Alkaline Solutions on Bone: Is Chemistry the Answer?

The treatment of corpses with extremely acid or basic liquids is sometimes performed in criminal contexts. A thorough characterization by chemical analysis may provide further help to macroscopic and microscopic analysis; 63 porcine bone samples were treated with solutions at different pH (1–14) for immersion periods up to 70 days, as well as in extremely acidic sulfuric acid solutions (9 M/18 M) and extremely basic sodium hydroxide. Inductively coupled optical emission spectrometry (ICP‐OES)/plasma mass spectrometry (ICP‐MS), Fourier transform infrared spectroscopy (FT‐IR), energy dispersive X‐ray analysis (EDX), X‐ray powder diffraction (XRPD), and scanning electron microscopy (SEM) showed that only the sulfuric acid solution 18 M was able to completely dissolve the sample. In addition, chemical analysis allowed to recognize the contact between bone and substances. Hydrated calcium sulfate arose from extreme pH. The possibility of detecting the presence of human material within the residual solution was demonstrated, especially with FT‐IR, ICP‐OES, and EDX.     

应用磁珠法提取陈旧骨骼DNA

目的探讨采用磁珠法提取陈旧骨骼DNA的可行性。方法取经土埋或室外暴露下存放1~5年不等的10根长骨,经水洗、刮净,钻取骨密质骨粉3g,应用EQ1000磁珠试剂盒提取DNA,经复合扩增,ABI 3130XL基因分析仪电泳分离,进行STR分型检测。结果 10根长骨均获得完整的STR分型,电泳图谱基线干净,除个别大片段基因座外,等位基因荧光信号分布均衡性较好。结论采用磁珠法提取陈旧骨骼的DNA,能满足分型要求,可在实际检案中选用。     

Preliminary Observations on the Ability of Hyperspectral Imaging to Provide Detection and Visualization of Bloodstain Patterns on Black Fabrics

Abstract: The analysis of bloodstain patterns can assist investigators in understanding the circumstances surrounding a violent crime. Bloodstains are routinely subjected to pattern analysis, which is inherently dependent upon the ability of the examiner to locate and visualize bloodstain patterns on items of evidence. Often, the ability to properly visualize bloodstain patterns is challenging, especially when the stain patterns occur on dark and/or patterned substrates. In this study, preliminary research was performed to better understand how near‐infrared reflectance hyperspectral imaging (HSI) could be used to observe bloodstain patterns on commonly encountered black fabrics. The ability of HSI to visualize latent bloodstains on several commonly encountered substrates is demonstrated. The images acquired through HSI are of sufficient quality to allow for differentiation between stains produced from an impact mechanism or a transfer mechanism. This study also serves as a proof of concept in the differentiation of multiple staining materials. Because of its ability to generate spectral data, the data provide a preliminary separation of stains where more than one type of stain existed.     

Interpreting Injury Mechanisms of Blunt Force Trauma from Butterfly Fracture Formation

According to biomechanics of fracture production during blunt impact, tubular bones are subject to compressive (impact site) and tensile (opposite impact site) forces; this causes bones to break in tension before compression, producing Y-shaped fracture patterns with breakaway (butterfly) fragments. In current forensic models, the side of the bone exhibiting the breakaway fragment is designated the impact side, with initial breakage occurring opposite. Fracture production and patterning of blunt impacts to 255 sheep femora were analyzed. Contra the existing model, only 60% of complete butterfly fractures exhibited impact side breakaway fragments. Although fractures initiated on the tension, nonimpact side, butterfly fragments formed on either compression or tension sides. Using newly defined breakaway fragment shape criteria, impact side was estimated with 98% accuracy for both complete and partial butterfly fractures. Furthermore, the results suggest that the impact site is the located on one of the Y-fracture's arms, not the butterfly fragment's center, as previously modeled.     

Comparison of DNA yield after long-term storage of Second World War bone samples

Sample storage is of paramount importance in forensic genetics laboratories since only optimal storage enables successful recovery of DNA from old bones that contain very low amount of severely degraded DNA. When identification of missing persons from skeletal remains is completed, bone sample is routinely stored at -20 °C for long-term storage for retesting in future, if necessary. After molecular genetic analyses of Slovenian Second World War (WWII) victims, small fragments of femurs were stored at -20 °C. Reduction in DNA recovery has been observed in frozen liquid DNA extracts by some authors and the goal of our study was to explore how freezing of bone samples affects the preservation of DNA. To achieve this goal, the difference in DNA yield in extracts obtained from WWII bones analyzed in 2009 (data from published paper) and DNA yield in extracts obtained from the same bones (piece sampled next to the one used in 2009) taken out of the freezer after long-term storage on -20 °C for 10 years was examined, using the same extraction method and the same quantification kit. Up to 100 ng DNA/g of bone powder was obtained from 57 WWII femurs and up to 31 ng DNA/g of bone powder from the same femurs investigated after long-term storage in this study. 0,5 g of bone powder was decalcified using full demineralization extraction method. The DNA was purified in a Biorobot EZ1 device (Qiagen) and DNA quantity determined with the Human Quantifiler kit (TFS). Statistical analysis showed significant difference in DNA yield in extracts obtained from WWII bones in 2009 and extracts obtained from the same bones stored at -20 °C after 10 years. As reported for frozen liquid DNA extracts, reduction in recovery of DNA was confirmed for frozen bone samples as well.     

Validity of the use of porcine bone in forensic cut mark studies

Porcine bone is often used as a substitute for human bone in forensic trauma studies, but little has been published on its comparative mechanical behavior. The factors affecting mechanical properties and therefore selection of bone models are complex and include the age of the animal at death, and physiological loading conditions, the latter being of particular relevance when using a quadrupedal animal as a human substitute. The regional variation in hardness of adult and infant porcine bones was investigated using Vickers’ indentation tests and compared to published data for human limb bones to relate differences to inherent genetic effects and loading influences, and to examine the validity of the porcine‐human model. Significant differences in hardness were observed both along and around the adult porcine humerus and femur, but no significant differences were found along the length of the infant bones. Significant differences were found between the forelimb and hindlimb, but only in the infant specimens. The hardness values for porcine adult cortical bone from the femur (52.23 ± 1.00 kg mm^?2) were comparable to those reported in the literature for adult human cortical bone from the fibula, ilium, and calcaneus. These data will help inform subject selection in terms of both species and bone type for use in future trauma studies.     

Criteria of personal identification by the hyoid bone

We studied 16 sizes of the hyoid bone (HB) from 158 corpses of known gender, Russians, who had died at the age over 15 years by the K. Miller et al program (1998) and 205 back projection electroroentgenograms of the neck of patients of the traumatological hospital in the town of Kirov. HB skeletization was made by the enzymatic drug enzyme. SPSS package and multidimentional statistics (regression and discriminant analyses) were used for data processing. It was established that HB is an informative object of forensic medical personal identification allowing determination of the body length and gender as well as anatomical, x-ray and osteometric individual features. Precision of human body length diagnosis by HB is 6.71-7.26 cm. Therefore, this method should be used in combination with the diagnosis by long tubular bones. Diagnosis of human gender by osteometric signs of HB based on one-dimentional discriminant analysis is feasible in 80-85%, based on canonical discriminant analysis--in 91.1% cases. The method of anatomic, osteometric and roentgenological assessment of HB individuality can be used in personal identification by intravital x-ray of cervical organs and in traumatic defects of HB. The methods may be applied in forensic medical practice in expert examination of body fragments or isolated bone fragments.