Estimating the Time of Deposition of Semen Traces using Fluorescence Protein-Lipid Oxidation Signatures.

In the forensic field, knowledge about the time of deposition of semen traces is extremely valuable to law enforcement agencies to assess the relevance of the traces and the validity of witness testimonies. However, currently, no method exists that is able to estimate the time of deposition of semen stains, due to the complex chemistry of the constituents and variation in degradation patterns. Here, we demonstrate a non-contact age estimation method to assess the time of deposition of semen stains using fluorescence spectroscopy. Protein-lipid oxidation reactions were monitored in semen stains over time using protein fluorescence and fluorescent oxidation product signatures to reveal distinctive aging patterns. On the basis of the relative amounts of these fluorescent products and the rate at which they are converted, successful age estimation was achieved up to a value of 16 days, with a median absolute error of 1.7 days. We demonstrate here a new tool that can fill the current gap in intelligence about the age of semen traces that has been challenging the forensic community worldwide.

Optimization of Vascular Casting for Three-Dimensional Fluorescence Cryo-Imaging of Collateral Vessels in the Ischemic Rat Hindlimb.

Development of collateral vessels, arteriogenesis, may protect against tissue ischemia, however, quantitative data on this process remain scarce. We have developed a technique for replicating the entire arterial network of ischemic rat hindlimbs in three dimensions (3D) based on vascular casting and automated sequential cryo-imaging. Various dilutions of Batson's No. 17 with methyl methacrylate were evaluated in healthy rats, with further protocol optimization in ischemic rats. Penetration of the resin into the vascular network greatly depended on dilution; the total length of casted vessels below 75 µm was 13-fold higher at 50% dilution compared with the 10% dilution. Dilutions of 25-30%, with transient clamping of the healthy iliac artery, were optimal for imaging the arterial network in unilateral ischemia. This protocol completely filled the lumina of small arterioles and collateral vessels. These appeared as thin anastomoses in healthy legs and increasingly larger vessels during ligation (median diameter 1 week: 63 µm, 4 weeks: 127 µm). The presented combination of quality casts with high-resolution cryo-imaging enables automated, detailed 3D analysis of collateral adaptation, which furthermore can be combined with co-registered 3D distributions of fluorescent molecular imaging markers reflecting biological activity or perfusion.

The impact of scaled boundary conditions on wall shear stress computations in atherosclerotic human coronary bifurcations.

The aim of this study was to determine if reliable patient-specific wall shear stress (WSS) can be computed when diameter-based scaling laws are used to impose the boundary conditions for computational fluid dynamics. This study focused on mildly diseased human coronary bifurcations since they are predilection sites for atherosclerosis. Eight patients scheduled for percutaneous coronary intervention were imaged with angiography. The velocity proximal and distal of a bifurcation was acquired with intravascular Doppler measurements. These measurements were used for inflow and outflow boundary conditions for the first set of WSS computations. For the second set of computations, absolute inflow and outflow ratios were derived from geometry-based scaling laws based on angiography data. Normalized WSS maps per segment were obtained by dividing the absolute WSS by the mean WSS value. Absolute and normalized WSS maps from the measured-approach and the scaled-approach were compared. A reasonable agreement was found between the measured and scaled inflows, with a median difference of 0.08 ml/s [-0.01; 0.20]. The measured and the scaled outflow ratios showed a good agreement: 1.5 percentage points [-19.0; 4.5]. Absolute WSS maps were sensitive to the inflow and outflow variations, and relatively large differences between the two approaches were observed. For normalized WSS maps, the results for the two approaches were equivalent. This study showed that normalized WSS can be obtained from angiography data alone by applying diameter-based scaling laws to define the boundary conditions. Caution should be taken when absolute WSS is assessed from computations using scaled boundary conditions.

Oxidation monitoring by fluorescence spectroscopy reveals the age of fingermarks.

No forensic method exists that can reliably estimate the age of fingermarks found at a crime scene. Information on time passed since fingermark deposition is desired as it can be used to distinguish between crime related and unrelated fingermarks and to support or refute statements made by the fingermark donors. We introduce a non-contact method that can estimate the age of fingermarks. Fingermarks were approached as protein-lipid mixtures and an age-estimation model was build based on the expected protein and lipid oxidation reactions. Two measures of oxidation are required from the fingermark to estimate its age: 1) the relative amount of fluorescent oxidation products 2) the rate at which these products are formed. Fluorescence spectroscopy was used to obtain these measures. We tested the method on 44 fingermarks and were able to estimate the age of 55% of the male fingermarks, up to three weeks old with an uncertainty of 1.9 days.

Topologic and Hemodynamic Characteristics of the Human Coronary Arterial Circulation.

BACKGROUND: Many processes contributing to the functional and structural regulation of the coronary circulation have been identified. A proper understanding of the complex interplay of these processes requires a quantitative systems approach that includes the complexity of the coronary network. The purpose of this study was to provide a detailed quantification of the branching characteristics and local hemodynamics of the human coronary circulation. METHODS: The coronary arteries of a human heart were filled post-mortem with fluorescent replica material. The frozen heart was alternately cut and block-face imaged using a high-resolution imaging cryomicrotome. From the resulting 3D reconstruction of the left coronary circulation, topological (node and loop characteristics), topographic (diameters and length of segments), and geometric (position) properties were analyzed, along with predictions of local hemodynamics (pressure and flow). RESULTS: The reconstructed left coronary tree consisted of 202,184 segments with diameters ranging from 30 µm to 4 mm. Most segments were between 100 µm and 1 mm long. The median segment length was similar for diameters ranging between 75 and 200 µm. 91% of the nodes were bifurcations. These bifurcations were more symmetric and less variable in smaller vessels. Most of the pressure drop occurred in vessels between 200 µm and 1 mm in diameter. Downstream conductance variability affected neither local pressure nor median local flow and added limited extra variation of local flow. The left coronary circulation perfused 358 cm3 of myocardium. Median perfused volume at a truncation level of 100 to 200 µm was 20 mm3 with a median perfusion of 5.6 ml/min/g and a high local heterogeneity. CONCLUSION: This study provides the branching characteristics and hemodynamic analysis of the left coronary arterial circulation of a human heart. The resulting model can be deployed for further hemodynamic studies at the whole organ and local level.

Endothelial shear stress estimation in the human carotid artery based on Womersley versus Poiseuille flow.

Endothelial shear stress (ESS) dynamics are a major determinant of atherosclerosis development. The frequently used Poiseuille method to estimate ESS dynamics has important limitations. Therefore, we investigated whether Womersley flow may provide a better alternative for estimation of ESS while requiring equally simple hemodynamic parameters. Common carotid blood flow, centerline velocity, lumen diameter and mean wall thickness (MWT) were measured with 3T-MRI in 45 subjects at three different occasions. Mean ESS and two measures of pulsatility [shear pulsatility index (SPI) and oscillatory shear index (OSI)] were estimated based on Poiseuille and Womersley flow and compared to the more complex velocity gradient modelling method. The association between ESS and MWT was tested with multiple linear regression analysis; interscan reproducibility was assessed using intraclass correlation coefficients (ICC). Mean ESS and pulsatility indices based on Womersley flow (ESSwq ß = -0.18, P = 0.04; SPIwq ß = 0.24, P = 0.02; OSIwq ß = 0.18, P = 0.045), showed equally good correlations with carotid MWT as the velocity gradient method (ESSvg ß = -0.23, P = 0.01; SPIvg ß = 0.21, P = 0.02; OSIvg ß = 0.07, P = 0.47). This in contrast to the Poiseuille flow method that only showed a good correlation for mean ESS (ESSpq ß = -0.18, P = 0.04; SPIpq ß = 0.14, P = 0.14; OSIpq ß = 0.04, P = 0.69). Womersley and Poiseuille methods had high intraclass correlation coefficients indicating good interscan reproducibility (both ICC = 0.84, 95% confidence interval 0.75-0.90). Estimation of ESS dynamics based on Womersley flow modelling is superior to Poiseuille flow modelling and has good interscan reproducibility.

3D Imaging of vascular networks for biophysical modeling of perfusion distribution within the heart.

One of the main determinants of perfusion distribution within an organ is the structure of its vascular network. Past studies were based on angiography or corrosion casting and lacked quantitative three dimensional, 3D, representation. Based on branching rules and other properties derived from such imaging, 3D vascular tree models were generated which were rather useful for generating and testing hypotheses on perfusion distribution in organs. Progress in advanced computational models for prediction of perfusion distribution has raised the need for more realistic representations of vascular trees with higher resolution. This paper presents an overview of the different methods developed over time for imaging and modeling the structure of vascular networks and perfusion distribution, with a focus on the heart. The strengths and limitations of these different techniques are discussed. Episcopic fluorescent imaging using a cryomicrotome is presently being developed in different laboratories. This technique is discussed in more detail, since it provides high-resolution 3D structural information that is important for the development and validation of biophysical models but also for studying the adaptations of vascular networks to diseases. An added advantage of this method being is the ability to measure local tissue perfusion. Clinically, indices for patient-specific coronary stenosis evaluation derived from vascular networks have been proposed and high-resolution noninvasive methods for perfusion distribution are in development. All these techniques depend on a proper representation of the relevant vascular network structures.