Quicklime and caustic soda water effects on a fresh cadaver.
Subject: Abortion (Risk factors)
Cadavers (Properties)
Women (Death of)
Lime (Chemical properties)
Author: Solla, Horacio E.
Pub Date: 06/22/2007
Publication: Name: The Forensic Examiner Publisher: American College of Forensic Examiners Audience: Professional Format: Magazine/Journal Subject: Health; Law; Science and technology Copyright: COPYRIGHT 2007 American College of Forensic Examiners ISSN: 1084-5569
Issue: Date: Summer, 2007 Source Volume: 16 Source Issue: 2
Product: Product Code: 3274000 Lime; 2812300 Caustic Soda NAICS Code: 32741 Lime Manufacturing; 325181 Alkalies and Chlorine Manufacturing SIC Code: 3274 Lime; 2812 Alkalies and chlorine
Geographic: Geographic Scope: Uruguay Geographic Code: 3URUG Uruguay
Accession Number: 165192826
Full Text: The leading cause of maternal death in Uruguay is caused by abortions made in uncertain conditions where the most elementary sanitary precautions are neglected. Daily, the poorest pregnant women in Uruguay allow unqualified individuals to perform abortions in clandestine places where extreme health risks exist. This article presents the case of Jane Doe, a 26-year-old Argentinean woman who died in Uruguay due to an acute hipovolemy caused by a secret illegal abortion made under unsanitary conditions. The midwife and her assistants who conducted the abortion sprinkled the cadaver with chemical agents commonly known as caustic soda water and quicklime to disfigure and/or destroy it completely before burying Jane Doe in the backyard of the midwife's property. This article's objective is to make a forensic contribution to what is known about quicklime and caustic soda water effects on fresh cadavers. This article is based on a case study where a fresh cadaver was subjected to such chemicals agents for about 1 year. It also describes changes on soft and bone tissues, hair, and dental pieces. The study showed that hair and long bones ephyphysis are more resistant to the effects of those chemical agents than other parts of the human body, like teeth and soft tissue. Using these body parts, DNA fingerprints analysis can help yield a positive identification of the human skeletal remains.

Introduction

This article illustrates the importance of a multidisciplinary plan of action in murder investigations. Murderers often dispose and destroy their victims' bodies using chemical agents. In the present case, the identification of the remains was very difficult due to the chemical agents Jane Doe's murderers applied to her cadaver. This case report on Jane Doe makes a forensic contribution regarding the effects of quicklime and caustic soda water on fresh human cadavers. This article describes several DNA techniques, although they are not the article's principal objective.

In Uruguay, many women die each year because of abortions done in unsanitary conditions by people unqualified to perform the operations. In most cases, midwives perform abortions without a medical attendant. Although many midwives conduct abortions regularly and are often considered experienced, they lack the training to safely perform the high-risk operations. Further adding to pregnant women's risks, having an abortion constitutes committing a crime in Uruguay's legal system unless it is done in sanitary conditions to safeguard the mother's physical integrity or the mother is especially poor and already has a lot of children. Evidence shows that the majority of abortions in Uruguay do not constitute an illegal activity. Nevertheless, each year approximately 33,000 women die from clandestine abortions. Almost half (47.9%) of the women who sought medical attention from Pereira Rossel Hospital for uterus infections resulting from abortions performed in unsanitary conditions died.

Jane Doe was a 26-year-old Argentian woman who, in April 2001, crossed the Rio de la Plata from Argentina to Uruguay to have an abortion. Abortions are cheaper in Uruguay than in Argentina, but are illegal in both countries. In her case, because of complications at a clinic, she was sent to a midwife's private house in a district in Montevideo's outskirts. When she arrived, the midwife and her assistants were waiting to proceed with the abortion that the clinic's properly trained doctors had deemed too dangerous.

As a last resort, several months after Jane Doe had left Argentina, her worried relatives brought her disappearance to the attention of the Immigration General Direction authorities. After investigating the case, the immigration authorities confirmed her entrance to Uruguayan territory at Colonia del Sacramento but could not find any record of her departure to Buenos Aires. After several investigations, the Interpol-Uruguay and local police found the midwife who performed the abortion. After her arrest, the midwife confessed to the crime and gave details of the event.

The victim died as a consequence of an acute hipovolemy resulting from a critical hemorrhage the midwife's incorrect surgical procedures caused during the high-risk abortion. The midwife also described what she and her assistants did with Jane Doe's cadaver and indicated where it was buried. The midwife and her assistants cut off several of Jane Doe's body parts and buried them in the midwife's backyard in the district of White Rocks near Montevideo. Before burying her, they sprinkled the body with quicklime and caustic soda water to destroy the cadaver so that if ever found, it would be difficult or impossible to identify.

The midwife's declarations to the local police and Interpol-Uruguay agents led the judge and the coroner to the buried corpse more than a year after Jane Doe's death. Investigators took several photographs with the remains in place. They also made a map to record location details and searched the surrounding area for artifacts and clues. It was helpful to know that no other bodies were lying around. No clues were found around the buried body's location. Next, Jane Doe's remains were removed. All the ground under and around the remains was sifted, but nothing was found. Then the body was carried to the Morgue Judicial of Montevideo City to be analyzed.

Chemistry of the Quicklime and Caustic Soda Water

The denomination of hydroxyl lime is a generic term that includes the oxides that, by means of industrial processes, imply the rock calcites with high calcium, dolomite, or magnesium content. This definition does not include limestone or other carbonates that, when simply crushed, are generally referred to as lime. The quicklime is the resulting product of the calcinations of limestone in furnaces with temperatures near 1000[degrees]C. This product is composed mainly of the oxide of calcium and magnesium. On the basis of chemical analyses, the quicklime can be divided in three types. First, quicklime of high calcium content is formed by about 95% calcium oxide (CaO) and less than 5% magnesium oxide (MgO), which demands a limestone rock of high calcium content and efficient calcinations. Second, quicklime dolomite is composed of around 40% magnesium oxide (MgO) and 57% calcium oxide (CaO), which requires a limestone rock of high purity in calcium and magnesium. Finally, quicklime magnesic is formed by 5 to 35% magnesium oxide and 60 to 90% calcium oxide (CaO), which is available in several common places in Uruguay. The quicklime has many applications, although the most common uses are environmental, such as the treatment of potable and industrial water, the treatment of the sewage system water and its moods, the treatment of the ground recovery water contaminated with hydrocarbons and reliable chemistries, the discoloration of several combustion gases, and the treatment of solid remainders. In the process of extinguishing the hydroxyl of calcium, the volume of dull lime [Ca (OH)2] expands to double the amount of initial quicklime (CaO), which is why it is also generally used to break rock or wood (Stryer, 1982).

The hydroxyl of sodium, better known as caustic soda water, is extremely corrosive and may cause serious burns when in contact with the skin. Thus, it is not rare to use it to disfigure and dissolve the soft parts of corpses that have been covertly buried.

From a forensic point of view, it is interesting to know the concrete changes in a fresh cadaver that has been exposed to quicklime and caustic soda water. Concerning the effects of quicklime on cadavers, Etxeberria (1988) found that while human bones are similar to incinerated bones (both being white and fragile), differences exist in their microscopic structures. Bones exposed to quicklime develop giant crystals formed by an accumulation of small crystals that are about 15 microns in length, while bones exposed to a temperature over 660[degrees]C develop crystals less than a micron in length.

Baud and Susini (1988) indicate that quicklime is corrosive and absorbs water from organic tissues, producing heat. However, this elevation of temperature lacks the capacity to burn human skeletal remains. Bones acquire a white color and a high degree of frailty but do not present any fractures or blacking effects like bones exposed directly to fire. Once quicklime comes into contact with the water of cadaver tissues, it is transformed into slaked lime that degrades the organic matter and the collagen of the bones, which explains the absence of organic matter in the bones and their lost elasticity.

Materials and Methods

Once in the Judicial Morgue of Montevideo, the local pathologist could not carry out an autopsy due the body's stage of decomposition. He sent the human remains to be analyzed by the resident forensic anthropologist, who wrote this article.

Anthropological Analysis of the Recovery Remains

The exhumed human remains were received within a black nylon bag and were at such a stage of decomposition that, from a macroscopic point of view, they did not have the appearance of a human body. Nevertheless, a lengthy analysis revealed that, indeed, they were human remains in an advanced stage of decomposition.

In order to determine that they were human remains, it was necessary to make certain sectioned cuts--specifically at the level of the thoracic region and the lumbar region where the ribs and the thoracic and lumbar column are appraised clearly by the anterior view. The head was not well preserved because several blows from a shovel had shattered it. Only a small part of the occipital bone was found, and no dental pieces were recovered. Therefore, no dental record comparison was possible.

The examined human remains did not have the usual foul odor of a decomposed body. After cleaning the bone remains with chlorinated water, abundant foam formation and a strong detergent odor were present. It was evident that the corpse had been exposed to some chemical substance to cause the body deterioration. Therefore, several samples were sent to the Judicial Morgue of Montevideo's Laboratory of Chemistry and Toxicology to be analyzed. Once analyzed, the presence of a strong caustic soda water and quicklime mixture was detected on the samples.

Using conserved fragments of long bones, especially the distal femur and proximal tibia, the stature of the victim was determined (Steele, 1970). The color of the skin did not undergo changes, which indicated a caucasic racial affinity. The study of the hipbones, composed by the ilium, ischium, and pubis, determined that the body belonged to a female (Genoves, 1959). The age at the time of the death came down to the study of the costo vertebral articulation joints (Iscan, Loth, & Wright, 1985) and the study of the pubic shymphysis (Gilbert & McKern, 1973). Also, there were remains of black hairs (about 20 centimeters long), black female underclothes, a sock, and a clear-colored blouse.

The body displayed clear signs of postmortem violence, especially at the skull, and there were several cuts by a sharpened instrument--like a big knife or an axe--in at least three sections. The first was at both arms, the second on the lumbar region, and a third was on both inferior members. There were no characteristics of individualization, except for the clothes found.

The results of the forensic anthropological studies show that those were the human remains of a white woman, approximately 167 cm tall, who was between 20 and 30 years old at the time of death.

Due to the fragmentation and decomposition of the cadaver, it was not possible to use anthropological methods of identification such as skull-photographic comparisons assisted by the computer, a radiographic-comparative method, or a dental comparison.

Nevertheless, from what arose from the investigations and from the declarations of the midwife, it was suspected that the cadaver belonged to Jane Doe. In fact, the sex, stature, hair color, and approximate age at the time of death matched the data of the missing Argentinian citizen. The DNA of the hair and the skeletal human remains, when compared with blood samples from Jane Doe's mother, gave positive results.

In the past decade, DNA analysis has been used as physical evidence in the investigations of crimes such as assault, sexual assault, and homicide for both psychological and legal reasons. Positive identification of a decedent is desired and may often be accomplished via the traditional scientific methods of comparative dental radiography, fingerprints matching, and osteological examination. However, in cases like this one, where there is an extensive mutilation of the soft tissues, disarticulation, destruction of key skeletal landmarks, and unavailable pre-mortem records, these traditional methods may not be useful.

Recent developments in DNA analysis and their application to forensic identification have revolutionized the possibility of identifying human remains. Crime labs around the world use more advanced and efficient methods of DNA analysis than restriction fragment length polymorphism (RFLP). However, today in Uruguay, the most available and widely applied genetic forensic identification or exclusion diagnosis is achieved using the established protocol for genomic DNA analysis, which is RFLP. This kind of analysis uses radio-labelled human-specific probes, which recognize and bind to restriction endonuclease-digested sample DNA that has been size separated and transferred onto a nylon membrane. Each probe detects the variable number of tandem repeats (VNTR) polymorphism within a hypervariable region of the human genome. The polymorphic nature of these VNTR is visualized as a band pattern on an autoradiograph and in this way demonstrates the capability of virtually individualizing human DNA.

To this point, the DNA identification currently employed in crime laboratories has been mainly confined to body fluids and soft tissue samples. Under laboratory conditions, blood and semen stains in a protected dry environment have yielded DNA still suitable for testing after decades. Teeth and hair may also serve as excellent sources of DNA and are biologically very stable. Many environmental conditions do not affect the ability to obtain a high molecular weight of human DNA from dental pulp or bones, which was discovered from examining bodies buried in various soil types. So in this case, human compact bone samples and hair remains were used as a source of DNA for RFLP analysis. The use of human bone and hair as a source for DNA typing is a relatively recent development in forensic science. The preservation of the DNA degradation of the sample is a limiting factor in DNA recovery. This common problem plagues this type of analysis. In this case, the chemical agents may limit the degradation of DNA post-mortem tissues. However, a strict correlation has not been shown between the degree of DNA degradation and the time since death when chemical agents are used to destroy a body. In this case, the DNA studies have shown the recovery of typeable DNA as late as 1 year post-mortem in the bones and hair samples analyzed.

DNA Analysis of Bones and Hair Samples

One section of the femoral ephyphysis and hair remains was removed immediately frozen at -80[degrees]C as a control set. The surface of the sample was sterilized with 70% ethanol wrapped in clean bench paper and fragmented with a mallet. Dehydration and desiccation were carried out with sequential washes of sterile deionised water, 100% ethanol, and ethyl ether shaken at 4[degrees]C for 20 minutes.

When dried, the sample was frozen in liquid nitrogen and ground to a fine powder using a blender fitted with a special stainless-steel specimen cup (Fisher Scientific). Two grams of bone powder were dissolved with 3 mL of 10 mM Tris (pH 8.0), 10 mM EDTA, 0.1 M NaCl, 2% SDS, and 0.5 mg/mL Proteinase K at 42[degrees]C for 16 hours in a shaking water bath. The Proteinase K concentration was then brought up to 1 mg/mL, and incubation continued 5 additional hours. The sample was centrifuged at 5,000 x g for 3 minutes at 4[degrees]C, and the supernatant was extracted with an equal volume of phenol/chloroform/isoamyl alcohol. The DNA was precipitated with 2.5 volumes of cold 100% ethanol. The DNA pellet was recovered by centrifugation at 12,000 x g for 12 minutes at 4[degrees]C and re-suspended in 10 mM Tris, 0.1 mM EDTA (pH 8.0).

DNA recovered from the bone samples was quantified by absorbance at 260 and 280 nm; agarose gel electrophoresis alongside known quantitative standards (15-500 ng DNA, Gibco BRL) and southern transblot of the agarose gel followed by hybridisation analysis using the P32 labelled, human-specific probe V1 (locus D17s79) (lifecodes). Isolated human DNA was detected by autoradiography. Transblots were also hybridised with two oligodeoxynucleotide probes, p11E and p13B, which were derived from conserved gene sequences for the 16S ribosomal RNA found in eubacteria.

RFLP analysis was performed according to the most current method described in the specialized literature. Briefly, the isolated DNA was digested with an excess of 5 times the restriction of endonuclease Hae III (Boehringer Mannheim) overnight at 37[degrees]C and extracted with phenol/chloroform/isoamyl alcohol, and the DNA was precipitated with ethanol. The DNA sample was then electrophoresed through a 1% agarose gel alongside size markers and a pre-digested positive control--DNA from the human cell line K562 (Lifecodes). After electrophoresis, the DNA was transblotted onto a Biodyne B membrane (Pall Biosupport) under alkaline conditions and covalently linked to the membrane in a CL-1000 ultraviolet cross-linker (UVP) at 254 nm with 120,000 uJ/cm2. Each membrane was hybridized with a human-specific DNA probe and DNA probed for the size markers, both labelled.

To compare the intensities of the signals detected amid the various time points for each treatment, the normalized values for all the fragments detected by each of the human-specific probes were pooled; this was based on the assumption that any particular exposure condition has the same (random) effect on each of the probed loci. The data were then analysed by repeat measures analysis of variance between groups (ANOVA).

Head hairs from different regions of the scalp were obtained during the anthropological analysis of the remains, stored in paper envelopes at an ambient temperature, and submitted for DNA analysis. Reference blood samples from Jane Doe's mother were collected in ethylenediamine tetraacetic acid (EDTA) tubes. In order to compare the DNA typing results, the same genetic markers in the victim's sample were analyzed.

DNA was isolated from 1 cm of the root portion from each of the hairs. The extraction of the reference head hairs and the blood samples were carried out independently at different times. Each hair was incubated overnight at 56[degrees]C in 400uL stain extraction buffer (10mM Tris, 10 mM EDTA, 100 mM C1, 39mM Dtt, 2% SDS) and 10uL Proteinase K (20mg/mL) were added. Then the samples were incubated for 2 more hours at 56[degrees]C. The solution was extracted with 500uL phenol-chloroform-isoamyalcohol (25:24:1) and subsequently extracted in 1 mL water saturated n-butamol to remove traces of phenol. The aqueous phase was then transferred to a Centricon 100 microconcentrator tube containing 1 mL sterile water. The volume was brought up to a total volume of 2 mL with sterile water, the sample reservoir was sealed with parafilm, and the tubes were subjected to centrifugation at 1000 g for 30 minutes. The DNA was recovered by back centrifugation at 1000 g for 5 minutes. The final sample volume was approximately 25-40 uL.

Nine nanograms of Human DNA could be extracted from the hair samples. Later, two nanograms from the sample were amplified and typed for HLA-DQ according to the protocol contained in the Amply-Type HLA-DQ alpha Amplification and Typing Kit. In order to conserve as much original evidentiary sample DNA as possible for further analysis, the genomic DNA from the HLA-DQA1 amplification product was purified, recovered, and subsequently amplified. The remaining portion of this sample has been stored at -20[degrees]C. Subsequently, the solution was transferred to a Centricon 100 microconcentrator tube containing 2 mL of sterile water and purified by removing the primers, dNTPs, and salts. The purified retentate containing the genomic DNA and the DQA1 amplification product was recovered by back centrifugation. The final sample volume was approximately 20-30 uL. Twenty uL were amplified in a 100 uL PCR in a Perkin Elmer Thermal Cyler 480 using the Ampli-Type PM PCR Amplification and Typing Kit and typed for five loci according to the manufacturers protocols. Briefly, the PCR was carried out in 50 uL reaction volumes in a Perkin Elmer 480 thermal cycler. Five uL of PCR product were mixed with 2.5uL 3 X STR loading dye and 2.5 uL of this mix were loaded onto a denaturing polycrylamide gel containing 7 M urea and 0.5 X Tris-Borate-EDTA buffer. Electrophoresis was carried out on an SA 32 Electrophoresis Apparatus. The conditions for electrophoresis were set at a constant power of 40 watts and electrophoresis was carried out at ambient temperature. Electrophoresis was stopped when the xylene cyanol dye migrated 6 cm from the anode (approximately 1 hour and 20 minutes). Allele designations were determined by comparison of the sample fragments with those of the allelic ladders supplied in the kit.

From the hair sample of Jane Doe, human DNA were isolated and amplified successfully and typed at nine genetic loci: HLADQA1, LDLR, GYPA, HBGG, D7S8, GC, HUMTH01, TPOX, CSF1P0. The DNA profiles obtained from Jane Doe's mother's DNA sample matched the DNA profiles obtained from Jane Doe's hair samples.

Discussion and Conclusions

DNA typing has been shown to be a powerful tool for identifying purposes. The detection of VNTR sequences by RFLP analysis can produce very high levels of discrimination, but it requires a minimum of 25 to 50 ng of relatively undegraded DNA. Genetic information from samples with degraded DNA or from samples that do not contain as much DNA may be obtained by amplification of the DNA using the PCR. PCR-based tests, including the amplification of the HLA DQA1 locus, the PM loci various VNTR loci, microsatellite typing, and mitochondrial DNA sequence analysis, have been successfully applied to the analysis of DNA extracted from various sources including bones and hairs.

Although identifying the remains with DNA technology was important, the principal objective of this article, from a forensic perspective, is to determine the effects of quicklime and caustic soda water on a fresh body. Therefore, in this case it is important to note and determine what parts of Jane Doe's cadaver had been affected, and to what degree, by the exposure to the indicated chemical agents.

In this case, the action of both chemical components used to disfigure and to dissolve the cadaver remains did not have the same effects on the soft tissue as they did on the bone tissue. Indeed, quicklime and caustic soda water destroyed the greater part of Jane Doe's soft tissue. However, although caustic soda water has the well-known destructive effect of burning the soft tissue of the human body, the most destructive effects were on the bone tissue and dental system. The bones that could be recovered displayed a blubbery aspect, were very fragile, and were destroyed very easily under minimum pressure. The dental pieces were disintegrated or were dissolved so that none that fit the typical anatomy of a human tooth could be identified (Der Boghosian, 1952).

Therefore, the identification of Jane Doe's remains had to be done by comparative studies of extracted DNA from hair remains and the few long bone fragments that could be recovered and blood samples from the victim's mother. It is also important to indicate that the hair did not show any destruction due to the exposure to the quicklime and caustic soda water, as did the soft tissue, bone tissue, and teeth.

The results of this study indicate 2 g of femoral cortical ephyphysis bone and a hair sample were sufficient to extract quality, typeable genomic DNA from a relatively fresh cadaver to a 1-year-old cadaver.

The main goal of this study is to establish human bone and hair as a valid and reliable source for use in DNA testing in forensic case work. The majority of the forensic cases in Uruguay can be identified by traditional anthropologic methods, yet in many cases, only estimations of age, race, sex, and stature may be made. The methods employed in this study are not only significant when there is direct ante-mortem evidence for DNA comparison, but may also be used in cases where the separation of co-mingled evidence is problematic, when the relationship between spatially disparate fragmentary remains is in question, or when first order living relatives of the victims are available for reverse-paternity testing.

Based on our initial analysis of the DNA extracted from various bone ephyphysis specimens, it is clear that bone can be used as a reliable source of DNA. An average of 73 ug per gram of bone was recovered from three separate specimens. Although the quality of the DNA may vary, RFLP analysis of the three bone specimens suggests that they yielded high quality restriction patterns. Exposure of these bone samples to various environmental insults indicates that the DNA is susceptible to the prolonged treatment of shallow burial and water immersion. After 6 months of exposure, the DNA demonstrated extensive degradation upon RFLP analysis. This was despite what appeared to be a good recovery of DNA material for both treatments at 6 months. This study consistently found that RFLP analysis required a large amount of DNA from these bone samples, even though in typical forensic RFLP analysis, only nanogram quantities of DNA are employed.

The sensitivity of the RFLP analysis depends largely on the preservation of the high molecular weight of human DNA, and contamination by microbial DNA in the bone extracts limits the sensitivity of this analysis. Conceivably, the sensitivity of the RFLP analysis may be enhanced by separating the human DNA from the microbial contamination prior to restriction of the sample, particularly in the case of surface deposits.

The results also support the contention that microbial activity and/or moisture in the soil may be important factors for DNA degradation in bones. The climatic conditions during the 1-year period that the body was buried were dry and cool loam; therefore, we consider that these conditions tend to retard the chemical action of quicklime and caustic soda water on flesh cadavers. Longer exposure time for autoradiography or increasing the amount of total DNA loaded may enhance the sensitivity of detection. Of the three forensic samples, as in this case, ephyphysis of the long bones was the most desirable source of DNA for RFLP analysis.

Thus, exposing a fresh cadaver to these two chemical agents causes an important deterioration in direct relation to the length of time it has been buried and the volume of chemical agents applied. In this case, a volume of quicklime and caustic soda water was applied to a young female of about 60 kg and 167 cm of stature until it covered the entire body, which was buried in a hole with a depth of about 50 cm, a length of about 2 m, and a breadth of about 50 cm.

The action of the quicklime and the caustic soda water on the human body is not homogeneous. These two chemical agents do not affect the soft tissue and the bone tissue the same way. The quicklime is almost harmless to bone tissue, because the tissue is protected by soft tissue. The caustic soda water, however, is very destructive to the bone tissue as well as the soft tissue. Several areas of the bone anatomy showed a greater resistance to the action of these chemical agents, like the proximal and distal region of long bones (see Figures 10-13). Finally, hair is particularly resistant to the action of these two chemical agents.

Keywords: forensic anthropology, abortions, quicklime and caustic soda water, DNA, identification

CE CREDIT

This article is approved by the following for continuing education credit:

(ACFEI) The American College of Forensic Examiners International provides this continuing education credit for Diplomates.

(CMI) The American College of Forensic Examiners International provides this continuing education credit for Certified Medical Investigators.

(CFN) The American College of Forensic Examiners International provides this continuing education credit for Certified Forensic Nurses.

(ADA) (CMI) The American College of Forensic Examiners International is an ADA CERP Recognized Provider

This article is dedicated to the memory of Mary Rita Quinteiro.

References

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Der Boghosian, B. (1952). Los dientes. Impresora Garcia Morales. Montevideo.

Etxeberria, F. (1988). Aspefctos macroscopicos del tejido oseo sometido al efecto de las altas temperaturas. Aportacion al estudio de las cremaciones. Revista Espanola de Medicina Legal, 19, 159-163.

Genoves, S. T. (1959). Diferencias sexuales en el hueso coxal. UNAM. Mexico City, D.F.: Publicaciones del Instituto de Historia. Primera serie. Num. 49.

Gilbert, B. M., & McKern, T. W. (1973, January). A method for aging the female Os pubis. American Journal of Physical Anthropology, 38(1), 31-38.

Iscan, M. Y., Loth, S. R., & Wright, R. K. (1985, July). Age estimation from the rib by phases analysis: White females. Journal of Forensic Science, 30(3), 853-863.

Steele, D. G. (1970). Estimation of stature from fragments of long limb bones. In Stewart, T. D. (Ed.), Personal identification in mass disasters (85-97). Washington, DC: National Museum of Natural History, Smithsonian Institution.

Stryer, L. (1982). Bioquimica. (2nd Ed.). Barcelona: Editorial Reverte.

Dr. Horacio E. Solla received his graduate degree in Anthropological Sciences in 1991 from the University of the Republic, Faculty of Humanities and Sciences, Uruguay. He has completed many post-graduate degree courses in forensic anthropology. He received a Certificate of Inclusion in the 2000 Outstanding Scholars of the 21st Century (first edition) in honor of an outstanding contribution to the field of forensic anthropology in Uruguay as the founder of forensic anthropology in his country. He also received a doctorate degree of Merit (PhD) from the International Biographical Institute at Cambridge, England (2001). He has published three books and more than 50 scientific articles.

Dr. Solla is a member of the Uruguayan Society of Forensic Sciences (1991), the Uruguayan Society of History of Medicine (1992), the American Academy of Forensic Sciences (1995), the American College of Forensic Examiners (2000), and the Spanish College of Forensic Experts (2002). He also was curator at the National Museum of Anthropology (1990-1992), assistant of physical anthropology at the University of the Republic, Montevideo (1989-1995), and a forensic anthropologist at the National Institute of Criminology (1992-1995). Presently, he has a full time position as a forensic anthropologist at the Judicial Morgue of Montevideo City (Secretary of Justice) where he has solved more than 600 forensic anthropology cases and identified more than 100 missing persons skeletal remains.
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