Tissues Of Victims Research Articles

A COMBINED APPROACH TO THE REMOVAL OF METALLIC FOREIGN BODIES FROM SOFT TISSUES

Introduction. The ongoing hostilities in Ukraine have led to a significant number of victims with gunshot wounds. A distinguishing characteristic of modern firearms and explosive weapons is the impact of projectiles that cause injuries in the form of fragments, bullets, secondary and tertiary shrapnel, and other foreign bodies (e.g., glass, plastic, soil, or clothing remnants) that remain embedded in the victim's tissues. Shrapnel wounds account for 56.7% of all combat injuries. The treatment of gunshot and bullet wounds focuses on the removal of foreign bodies and the restoration of the anatomical and physiological integrity of the affected tissues. However, there is currently no standardized approach to the removal of such foreign bodies. The objective of this study was to enhance outcomes in the removal of metallic foreign bodies by employing a combined method during repeated surgical wound treatment. Participants and methods. The study involved two research groups. The first group (Group I) consisted of 191 wounded individuals whose metal fragments were removed using the traditional method with magnetic instruments. The second group (Group II) included 173 patients whose foreign bodies were removed using a combination of a mobile X-ray unit and ultrasound guidance. Results. The findings demonstrated that the combined method for removing foreign bodies offers significant advantages over the traditional approach. The combined method increased the removal efficiency by 20.6%. Intraoperative complications were 5.2 times less frequent in Group II compared to Group I (1.7% vs. 8.9%). Postoperative pain levels, measured using the Visual Analog Scale (VAS), were significantly lower in Group II (4.5 ± 0.32) compared to Group I (7.5 ± 0.3). Additionally, the combined method effectively minimized the risk of inflammatory processes and reduced the incidence of postoperative complications. These results highlight the combined method as a superior approach for the removal of metallic foreign bodies, improving both surgical outcomes and patient recovery.

Bioactive peptides (cryptides) obtained by Bothrops jararaca serine peptidases action on myoglobin

Serine peptidases and metallopeptidases are the primary toxins found in Bothrops snakes venoms, which act on proteins in the tissues of victims or prey, and release of peptides formed through proteolytic activity. Various studies have indicated that these peptides, released by the proteolytic activity of heterologous enzymes, generate molecules with unidentified functions, referred to as cryptids. To address this, we purified serine peptidases from Bothrops jararaca venom using molecular exclusion chromatography and then incubated them with the endogenous substrate myoglobin. As a control, we also incubated the substrate with trypsin. The resulting proteolytic fragments were analyzed, separated, and collected via HPLC. These fractions were then tested on cell cultures, the active fractions were sequenced (ALELFR and TGHPETLEK) and synthesized. After confirming their activity, the peptides underwent sequencing and synthesis for additional cell tests, including the increase of cell viability, cycle phases, proliferation, signaling, growth kinetics, angiogenesis, and migration. The results revealed that the synthesized peptides exhibited cellular repair properties, suggesting a potential role in tissue repair in the range of 0.05–5 μ M. Additionally, the effects of fragments resulting from myoglobin degradation isolated (ALELFR and TGHPETLEK) revealed a regenerative action on tissue.

Functional Ultrastructure of Hymenopteran Stingers: Devastating Spear or Delicate Syringe.

In this study, we tested the hypothesis that a micro-serrated edge on the honey bee Apis mellifera stinger tip serves as a tool for more intensive crushing of cell membranes in the victim's tissues. This could have mechanical consequences as well as initiate metabolic pathways linked to cell membrane breakdown (e.g., production of biogenic amines). Accordingly, we found that hymenopteran species that use their stingers as an offensive or defensive weapon to do as much damage to the victim's body as possible had this cuticular microstructure. In parasitic hymenopterans, on the other hand, this structure was missing, as stingers are solely used to delicately transport venom to the victim's body in order to do little mechanical harm. We also demonstrated that the stinger lancets of the honey bee A. mellifera are living organs with sensilla innervated by sensory neurons and containing other essential tissues, rather than mere cuticular structures.

Case report: Forensic 3D-match of hair brush and scalp abrasions revealing dynamic brush deformation

Abstract Forensic injury interpretation can be greatly enhanced by the use of 3D-surface documentation, also in conjunction with full body scans. For static juxtapositioning, a rigid shape transfer from weapon or injury causing object to the victim's skin or bone will be assumed for patterned injuries, upon which an object's shape directly explains the shape of an injury. Forensic pathology expertise may be essential in reading the resulting 3D shape match attempts, particularly when in due process of injury causation, deformation of either the object/weapon or victim's tissues occurred, causing a dynamic rather than rigid shape transfer. In this case, a 3D match using manual forensic juxtapositioning of punctate and elongated striate forehead skin abrasions and a hairbrush yielded a partial but not perfect match. Qualitative medicolegal appreciation of the partial match result clearly indicated centrifugal brush bristle deformation after impact, causing a centrifugal abrasion pattern, thus having dynamically extended an initially stationary punctate abrasion and bristle end shape match. This was corroborated by both the elasticity of the bristles and the brush frame fracture. Presence and location of the fracture indicated a relatively high force between the brush handle on one hand and the bristles on the other. This case shows how the application of targeted forensic pathology interpretation can elucidate what technically only is a partial 3D match and add re-constructive meaning. Generally, more research is needed in establishing just how technical sciences and specific medicolegal expertise are best integrated.

The Great Irish Famine: Identifying Starvation in the Tissues of Victims Using Stable Isotope Analysis of Bone and Incremental Dentine Collagen.

The major components of human diet both past and present may be estimated by measuring the carbon and nitrogen isotope ratios (δ13C and δ15N) of the collagenous proteins in bone and tooth dentine. However, the results from these two tissues differ substantially: bone collagen records a multi-year average whilst primary dentine records and retains time-bound isotope ratios deriving from the period of tooth development. Recent studies harnessing a sub-annual temporal sampling resolution have shed new light on the individual dietary histories of our ancestors by identifying unexpected radical short-term dietary changes, the duration of breastfeeding and migration where dietary change occurs, and by raising questions regarding factors other than diet that may impact on δ13C and δ15N values. Here we show that the dentine δ13C and δ15N profiles of workhouse inmates dating from the Great Irish Famine of the 19th century not only record the expected dietary change from C3 potatoes to C4 maize, but when used together they also document prolonged nutritional and other physiological stress resulting from insufficient sustenance. In the adults, the influence of the maize-based diet is seen in the δ13C difference between dentine (formed in childhood) and rib (representing an average from the last few years of life). The demonstrated effects of stress on the δ13C and δ15N values will have an impact on the interpretations of diet in past populations even in slow-turnover tissues such as compact bone. This technique also has applicability in the investigation of modern children subject to nutritional distress where hair and nails are unavailable or do not record an adequate period of time.

Application of LC-MS/MS in Identification of Toxin in the Causative Gastropod and Victim Samples

Food paralytic poisoning incident was detected in the gastropod Nassariidae Niotha clathrata and Zeuxis scalaris occurred in December 2012 in Kaohsiung, Taiwan. The toxin was purified from each remain of cooked gastropod and victim tissues by ultrafiltration using a YM-3 membrane, followed by C18 cartridge column purification. Based on analyses by LC-MS/MS, the causative agent of gastropod food poisoning was identified as tetrodotoxin. The toxicity of remain of cooked sample was 245 MU/g (1MU = 0.178 μg TTX) for N. clathrata and 203 MU/g for Z. scalaris. The blood and excrements of victim was shown to contain TTX of 0.13 and 0.18 μg/g, respectively. There was positive between results from LC-MS/MS and mouse bioassay in the causative gastropod. The LC-MS/MS method is suitable for rapid screening of TTX.

Complexes of ferriheme nitrophorin 4 with low-molecular weight thiol(ate)s occurring in blood plasma

Nitrophorins are proteins occurring in the saliva of the blood-sucking insect Rhodnius prolixus to carry NO as a vasodilator and blood-coagulation inhibitor into the victim's tissue. It was suggested that the rate of NO release can be enhanced by the blood-plasma component l-cysteine [J.M.C.Ribeiro, Insect Biochem. Mol. Biol. 26 (1996) 899–905]. However, the mechanism of the reaction is not clear. In the attempt to exploit the reaction in detail, complexes of nitrophorin 4 (NP4) with the thiols 2-mercaptoethanol, l-cysteine, and l-homocysteine and with HS− were formed and characterized under anaerobic conditions using absorption spectroscopy, X-ray crystallography, and EPR spectroscopy. In contrast to met-myoglobin, which is reduced by l-cysteine, all four compounds form low-spin FeIII complexes with NP4. The weak equilibration constants (167–5200M−1) neither support significant complexation nor the simple displacement of NO in vivo. Both amino acid based thiols form additional H-bonds with side chains of the heme pocket entry. Glutathione and l-methionine did not form a complex, indicating the specificity of the complexes with l-cysteine and l-homocysteine. Continuous wave EPR spectroscopy reveals the simultaneous existence of three low-spin systems in each case that are attributed to various protonation and/or conformational stages in the heme pocket. Electron nuclear double resonance (ENDOR) spectroscopy demonstrates that the thiol sulfurs are, at least in part, protonated. Overall, the results not only demonstrate the good accessibility of the NP4 heme center by biologically relevant thiols, but also represent the first structural characterization of a ferriheme protein in complex with l-cysteine l-homocysteine.

Bond or Cage Effect: How Nitrophorins Transport and Release Nitric Oxide

Most blood-sucking insects possess salivary proteins which, upon injection into the victim's tissue, help them improve their feeding. One group of these salivary proteins takes advantage of the vasodilator properties of NO to perform this task. These proteins are the so-called nitrophorins (NPs). NPs are heme proteins that store and transport NO, which, when released in the victim's tissue, produces vasodilation and inhibition of blood coagulation. It has been proposed that NO binds tightly to NP at a low pH of around 5.6 and that once NPs are injected in the victims tissue, at a pH of approximately 7.4, a conformational change occurs which lowers NO affinity, allowing it to be released. In this work we have studied the NO release mechanism of NP4 at a molecular level using state of the art computer simulation techniques. We have used molecular dynamics (MD) simulations to study NP4 conformational dynamics at both pH values 5.6 and 7.4 and computed the corresponding free energy profile for NO release using a multiple steering molecular dynamics scheme. We also have used hybrid quantum mechanical/molecular mechanics (QM/MM) techniques to analyze the heme-NO structure and the Fe-NO bond strength in the different NP4 conformations. Our results provide the molecular basis to explain that NO escape from NP4 is determined by differential NO migration rates and not by a difference in the Fe-NO bond strength. In contrast to most heme proteins that control ligand affinity by modulating the bond strength to the iron, NP4 has evolved a cage mechanism that traps the NO at low pH and releases it upon cage opening when the pH rises.

Protein Functional Cycle Viewed at Atomic Resolution: Conformational Change and Mobility in Nitrophorin 4 as a Function of pH and NO Binding,

The blood-sucking insect Rhodnius prolixus uses nitrophorin 4, a heme protein, to deliver nitric oxide (NO) to a victim, causing vasodilation and improved feeding. Binding of NO occurs at a ferric heme and is modulated by pH. NO binding at lower pH induces a large conformational change involving loops A-B and G-H that leads to distal pocket desolvation and protection of the nitrosyl heme complex. We have determined the crystal structures of Rhodnius nitrophorin 4 to ultrahigh resolution in four functional states: +/-NO at pH = 7.4 and +/-NO at pH = 5.6. The structure with NO at pH 7.4 (1.08 A) is newly determined while the other complexes have been modeled to resolutions much greater than previously reported (1.0-0.85 A). The ultrahigh resolution allowed us to resolve multiple conformers in binding-site loops, leading to a detailed description of the dynamics involved with storing NO in the insect salivary gland at low pH, and releasing NO in response to the increased pH of a victim's tissue. Strikingly, features for both the "open" and "closed" conformers exist under all conditions, suggesting that the flexible loops can transition with relative ease between conformational states. Yet, release of NO from rNP4 is much slower than found for other ferric heme proteins. The structures suggest that highly mobile loops can limit diffusion of diatomic molecules into and out of a protein cavity, a result with implications for the role of protein dynamics in function.

Role of binding site loops in controlling nitric oxide release: structure and kinetics of mutant forms of nitrophorin 4.

Nitrophorins are ferric heme proteins that transport nitric oxide (NO) from blood-sucking insects to victims. NO binding is tighter at lower pH values, as found in the insect salivary gland, and weaker at the pH of the victim's tissue, facilitating NO release and subsequent vasodilation. Previous structural analyses of nitrophorin 4 (NP4) from Rhodnius prolixus revealed a substantial NO-induced conformational change involving the A-B and G-H loops, which rearrange to desolvate the distal pocket and pack nonpolar residues against the heme-ligated NO. Previous kinetic analyses revealed a slow, biphasic, and pH-dependent NO release, which was proposed to be associated with loop movements. In this study, we created NP4 mutants D30A and D30N (A-B loop), D129A/L130A (G-H loop), and T121V (distal pocket). Eight crystal structures were determined, including complexes with NO, NH(3), and imidazole, to resolutions as high as 1.0 A. The NO-induced conformational change is largely abolished in the loop mutants, but retained in T121V. Kinetic analyses using stopped-flow spectroscopy revealed the pH dependence for NO release is eliminated for D129A/L130A, considerably reduced for D30A and D30N, but retained for T121V. NO association rates were increased 2-5-fold for T121V, but were unchanged in the loop mutants. Taken together, our findings demonstrate that the pH dependency for NO release is linked to loop dynamics and that solvent reorganization is apparently rate-limiting for formation of the initial iron-nitrosyl bond. Interestingly, the multiphasic kinetic behavior of rNPs was not affected by mutations, and its cause remains unclear.

Diatom analysis in victim's tissues as an indicator of the site of drowning.

The diagnosis of drowning is one of the most difficult in forensic pathology and previously we proposed criteria for a positive tissue analysis according to the qualitative and quantitative diatom investigations. In the positive cases, we studied the reliability of determining the site of drowning by comparing the diatom taxa found in the lung samples with those of the water samples or in the absence of these samples with the results of the water diatom monitoring programme set up in our region. In this study, we present two series of cases, the first is one of 20 corpses who died from accidental or suicidal drowning with known drowning site, and the second of 20 corpses for which the drowning site was unknown. The results showed that a concordance of the abundance of the diatom taxa in tissues compared to the site of drowning and their distribution relative to one another was 65% in the group where the site of drowning was known and 35% in the other group. The concordance of the individual distribution in the lungs of water diatom taxa may be an interesting method to guide the investigations for determining the site of drowning. The two limiting factors are the concentration of diatoms in the lungs and the development of a river monitoring programme in the district of the study.

Dose estimation by ESR spectroscopy at a fatal radiation accident

In Norway, 1982, a fatal radiation accident occurred at a radioactive cobalt source used for radiation processing. There was a lack of realistic estimates on the victim's tissue or body doses up to his death. GSF was able to provide the first reliable dose values, upon request from Norwegian colleagues. Dosimetry was performed by ESR spectroscopy of long-lived radiation-induced radicals that could be measured in the heart tablets of the victim. The concentration of the free radicals proved, by proper calibration, to be proportional to dose. Procedures and dose results are reported; comparison is made with data from literature dealing with the same subject and also using TL as well as ESR. For biological relevance, the doses as determined from the tablets are converted into whole-body doses.