Luminescence of thermally altered human skeletal remains.

Literature on luminescent properties of thermally altered human remains is scarce and contradictory. Therefore, the luminescence of heated bone was systemically reinvestigated. A heating experiment was conducted on fresh human bone, in two different media, and cremated human remains were recovered from a modern crematory. Luminescence was excited with light sources within the range of 350 to 560 nm. The excitation light was filtered out by using different long pass filters, and the luminescence was analysed by means of a scoring method. The results show that temperature, duration and surrounding medium determine the observed emission intensity and bandwidth. It is concluded that the luminescent characteristic of bone can be useful for identifying thermally altered human remains in a difficult context as well as yield information on the perimortem and postmortem events.

Temperature estimations of heated bone: A questionnaire-based study of accuracy and precision of interpretation of bone colour by forensic and physical anthropologists.

The colour of thermally altered bone, recovered from archaeological and forensic contexts, is related to the temperature(s) to which it was exposed. As it is heated bone changes in colour from ivory white, to brown and black, to different shades of grey and chalky white. It should be possible to estimate exposure temperature based on visually observable changes in colour. In forensic casework the temperature that human remains have been subjected to can reveal information about the existence and nature of foul play. Therefore, it is important to understand the accuracy and precision of visual methods of temperature estimation. Twenty-eight forensic and/or physical anthropologists estimated the temperature that fourteen bone samples had been subjected to based only on their colour via an online questionnaire. Bone samples shown in the questionnaire ranged from unheated to having been heated at 1200°C. Respondents were given two options to base their estimates on, resulting in a multiple response analysis. The results suggest it is difficult to identify the correct temperature range based solely on colour. Most respondents felt confident enough to opt for a single option, which may have contributed to a relatively high number of incorrect estimates. Low accuracy and precision were found for most of the temperature ranges, especially in the lower and middle categories. This study demonstrates that caution should be taken in the reliance upon temperature estimates of thermally induced colour changes in bone and the need for further research and improved methods.

Analysis of morphological characteristics and expression levels of extracellular matrix proteins in skin wounds to determine wound age in living subjects in forensic medicine.

OBJECTIVE: Wound age determination in living subjects is important in routine diagnostics in forensic medicine. Macroscopical description of a wound to determine wound age however is inadequate. The aim of this study was to assess whether it would be feasible to determine wound age via analysis of morphological characteristics and extracellular matrix proteins in skin biopsies of living subjects referred to a forensic outpatient clinic. METHODS: Skin biopsies (n=101), representing the border area of the wound, were taken from skin injuries of known wound age (range: 4.5h-25 days) in living subjects. All biopsies were analyzed for 3 morphological features (ulceration, parakeratosis and hemorrhage) and 3 extracellular matrix markers (collagen III, collagen IV and α-SMA). For quantification, biopsies were subdivided in 4 different timeframes: 0.2-2 days, 2-4 days, 4-10 days and 10-25 days old wounds. Subsequently, a probability scoring system was developed. RESULTS: For hemorrhage, collagen III, collagen IV and α-SMA expression no relation with wound age was found. Ulceration was only found in wounds of 0.2-2, 2-4 and 4-10 days old, implying that the probability that a wound was more than 10 days old in case of ulceration is equal to 0%. Also parakeratosis was almost exclusively found in wounds of 0.2-2, 2-4 and 4-10 days old, except for one case with a wound age of 15 days old. The probability scoring system of all analyzed markers, as depicted above, however can be used to calculate individual wound age probabilities in biopsies of skin wounds of living subjects. CONCLUSIONS: We have developed a probability scoring system, analysing morphological characteristics and extracellular matrix proteins in superficial skin biopsies of wounds in living subjects that can be applied in forensic medicine for wound age determination.

Analysis of inflammatory cells and mediators in skin wound biopsies to determine wound age in living subjects in forensic medicine.

OBJECTIVE: In forensic medicine it is important to determine the age of skin wounds in living subjects. The aim of this study was to assess whether analysis of inflammatory cells and inflammatory mediators in skin biopsies of wounds from living subjects could improve wound age determination. METHODS: Biopsies (n=101), representing the superficial border area of a skin wound, were taken from skin injuries of known wound age (range: 4.5 hours to 25 days) of living subjects. All biopsies were analyzed for 3 inflammatory cell markers (MPO, CD45 and CD68) and 4 inflammatory mediators (MIP-1, IL-8, CML and vitronectin). For quantification, biopsies were subdivided in 4 different timeframes: 0.2-2 days, 2-4 days, 4-10 days and 10-25 days old wounds. Subsequently, a probability scoring system was developed. RESULTS: MPO, CD45, MIP-1, IL-8 (inflammatory cell markers) and N(epsilon)-(carboxymethyl)lysine (CML) positivity were maximal in wounds of 0.2-2 days old and then decreased in time. Remarkably, CD45, CD68 and CML showed a minor but non-significant increase again in 10-25 days old wounds. MPO and CD68 positivity was significantly lower in 4-25 days old wounds compared to 0.2-4 days old wounds. MPO positivity was also significantly lower in 10-25 days old wounds compared to 0.2-10 days old wounds. For CD45, MIP-1, IL-8 and CML no significant differences between the age groups were found. In case of vitronectin positivity in the extravasate or when the number of MIP-1 or IL-8-positive cells was more than 10 cells/mm(2) the probability that a wound was more than 10 days old was 0%. A probability scoring system of all analyzed markers can be used to calculate individual wound age probabilities in biopsies of skin wounds of living subjects. CONCLUSIONS: We have developed a probability scoring system of inflammatory cells and mediators that can be used to determine wound age in skin biopsies of living subjects.

An evaluation of the time of discovery of fetal malformations by an indication-based system for ordering obstetric ultrasound.

Circumstances of detection of 570 structural abnormalities in 364 fetuses were reviewed to determine whether referral for obstetric ultrasound according to specific indications resulted in late detection of abnormal fetuses and whether earlier detection might have changed pregnancy outcomes. A system of indication-based obstetric ultrasound discovered 124 abnormal fetuses (34%) at 22 weeks or less and 240 (66%) at 23 weeks or more. Most fetal abnormalities found at 23 weeks or more were probably detectable earlier, because the pattern of abnormalities discovered was reasonably similar in the two groups. Discovery of abnormal fetuses at 22 weeks or less was associated with a 67% termination rate and an 11% postnatal survival rate, whereas discovery at 23 weeks or more was associated with a 14% termination rate and a 51% postnatal survival rate. For fetal abnormalities not detected until 23 weeks or more, the indications that led to detection were present earlier in only 28%, and any indications were present earlier in only 44%. This study raises serious concern about the ability of the indication-based obstetric ultrasound system commonly used in the United States to detect fetal abnormalities before therapeutic options become limited. Evaluation of alternative systems for timing of obstetric ultrasound appears to be warranted.

Fetal malformations commonly detectable on obstetric ultrasound.

A review of 364 fetuses with a total of 570 malformations discovered on ultrasound throughout the Emanuel Hospital referral pattern from 1978 to 1987 was compared to a previous multicenter study of 50,282 children from 1959 to 1965 that found minor or major congenital malformations in 6.5% of children, for a rate of 8.8 malformations per 100 children. The comparison was made to determine which of the malformations reported in the earlier study were commonly detectable on obstetric ultrasound as performed throughout the referral area and to determine what the combined prevalence of those malformations was within the general population. For the comparison, the number of patients undergoing obstetric ultrasound throughout our referral area was estimated from the number of anencephalic fetuses found. The relative prevalences of all malformations in the two groups were then determined by extrapolation. The malformations commonly detectable on ultrasound according to that comparison were then assigned the prevalences obtained from the earlier clinical study. According to this analysis, 0.7-0.8% of fetuses have major malformations commonly detectable on ultrasound, for a rate of 1.2 malformations per 100 fetuses. That represents about 13% of all malformations and about 27% of major malformations. If cardiovascular abnormalities, cleft lip and clubfoot were usually detected, the rate of malformations considered commonly detectable would increase to 2.7 per 100 fetuses. That would represent about 31% of all malformations and about 63% of major ones.

A new method to determine wound age in early vital skin injuries: a probability scoring system using expression levels of Fibronectin, CD62p and Factor VIII in wound hemorrhage.

PURPOSE OF THE STUDY: In forensic autopsies it is important to determine the age of early vital skin wounds as accurate as possible. In addition to inflammation, coagulation is also induced in vital wounds. Analysis of blood coagulation markers in wound hemorrhage could therefore be an important additional discriminating factor in wound age determination. The aim of this study was to develop a wound age probability scoring system, based on the immunohistochemical expression levels of Fibronectin, CD62p and Factor VIII in wound hemorrhage. METHODS: Tissue samples of (A) non injured control skin (n=383), and samples of mechanically induced skin injuries of known wound age, (B) injuries inflicted shortly before death (up to a few minutes old) (n=382), and (C) injuries inflicted 15-30 min before death (n=42) were obtained at autopsy in order to validate wound age estimation. Tissue slides were stained for Fibronectin, CD62p and Factor VIII and were subsequently scored for staining intensity (IHC score) in wound hemorrhage (1=minor, 2=moderate, 3=strong positive). Finally, probability scores of these markers were calculated. RESULTS: In at most 14% of the non-injured control samples, hemorrhage was found, with mean±standard deviation IHC scores of 0.1±0.4, 0.2±0.4 and 0.2±0.5 for Fibronectin, CD62p, and Factor VIII, respectively. Expression of these markers significantly increased to mean IHC scores 1.4±0.8 (Fibronectin), 1.2±0.6 (CD62p), and 1.6±0.8 (Factor VIII) in wounds inflicted shortly before death (few minutes old) and to 2.6±0.5 (Fibronectin), 2.5±0.6 (CD62p), and 2.8±0.4 (Factor VIII) in 15-30 min old wounds. The probabilities that a wound was non vital in case of an IH score 0 were 87%, 88% and 90% for Fibronectin, CD62p, and Factor VIII, respectively. In case of an IHC score 1 or 2, the probabilities that a wound was a few minutes old were 82/90%, 82/83% and 72/93%. Finally, in case of an IHC score 3, the probabilities that a wound was 15-30 min old were 65%, 76% and 55%. CONCLUSIONS: Based on the expression of Fibronectin, CD62p and Factor VIII in wound hemorrhage, we developed a probability scoring system that can be used in forensic autopsies to improve wound age estimation in early skin injuries.

The effect of glucocorticoids on the maturation of premature lung membranes. Preventing the respiratory distress syndrome by glucocorticoids.

The effect of glucocorticoid on the maturation of premature lung membranes was studied in 121 premature infants by administering variable dosages of Decadron to the 114 mothers prior to delivery. The results were compared with findings in a group of 390 infants born in the same hospital during this study. Administration of all three test doses, 8, 16, and 24 mg., significantly decreased the incidence of RDS in all gestational age and birth weight categories. For infants less than 32 weeks, the incidence was decreased from 75 to 46.2%; those 32 to 36 weeks, from 58 to 20.2%; and in those older than 36 weeks, from 24.4 to 0 per cent. The incidence in infants less than 1,000 grams was reduced from 100 to 71.5%; 1,000 to 1,500 grams, from 67.4 to 21.6%; 1,500 to 2,000 grams, from 52.3 to 22.6%; and in heavier than 2,000 grams, from 38.1 to 13.4%. The results also showed that glucocorticoid does not significantly reduce RDS if administered less than 24 hours prior to delivery. The incidence is reduced more than 50% if administered more than 24 hours prior to delivery.

Improved fluorescence polarization assay for use in evaluating fetal lung maturity. I. Development of the assay procedure.

We describe a fluorescence polarization assay for use in predicting fetal lung maturity, which is suitable for the TDx Analyzer (an automated fluorescence polarimeter). The assay requires 0.5 mL of amniotic fluid and approximately 30 min, and involves the fluorophore 1-palmitoyl-2(6-[(7-nitro-2, 1, 3-benzoxadiazol-4-yl)amino]caproyl) phosphatidylcholine. Reproducible polarization measurements depend on proper regulation of incubation time and temperature, but variations in the concentrations of fluorescent probe and amniotic fluid have little effect on measured polarization and therefore little effect on assay precision. Working solutions of the fluorescent probe are stable for at least nine months when stored at -20 degrees C and pH 5. Interferences include erythrocytes, serum, bilirubin, meconium, and lidocaine.

Improved fluorescence polarization assay for use in evaluating fetal lung maturity. II. Analytical evaluation and comparison with the lecithin/sphingomyelin ratio.

We assessed the analytical performance of an improved fluorescence polarization assay for use in evaluating fetal lung maturity and compared results with the lecithin/sphingomyelin ratio. During a three-month period 150 patients' samples were assayed by clinical laboratory personnel with both techniques. Values for the lecithin/sphingomyelin ratio correlate closely with net fluorescence polarization values (r = -0.85), less closely with net fluorescence intensity (r = 0.65). Background fluorescence intensity and polarization varied widely, indicating a need to correct measurements for endogenous fluorescence. Net fluorescence polarization values have a CV of 0.32% within-run, 1.07% between-day. A comparison of two amniotic fluid centrifugation procedures showed no significant difference in such values. For both methods, however, such values are slightly but significantly higher than those obtained for amniotic fluids without prior centrifugation. Short-term storage (less than 30 days) of uncentrifuged amniotic fluid samples at -20 degrees C does not significantly affect results.

Improved fluorescence polarization assay for use in evaluating fetal lung maturity. III. Retrospective clinical evaluation and comparison with the lecithin/sphingomyelin ratio.

We clinically evaluated, retrospectively, our improved fluorescence polarization assay for fetal lung maturity. The procedure requires 0.5 mL of amniotic fluid and a standard clinical laboratory fluorescence polarimeter (TDx Analyzer, Abbott Laboratories). We measured the L/S ratios for 93 freshly collected amniotic fluids, uncontaminated with blood or meconium, collected within three days of delivery. The fluids were stored frozen for eight to 32 months, then thawed and assayed for net fluorescence polarization. Fourteen of the infants developed respiratory distress syndrome; five, transient tachypnea of the newborn; and 74, no respiratory distress. The polarization assay and lecithin/sphingomyelin ratio had equivalent receiver operating characteristic curves, indicating no difference in their clinical performance. Although a prospective study with fresh amniotic fluid specimens will be necessary to establish a definitive reference range, the present study shows that this assay can be used to rapidly predict fetal lung maturity.