Background
A precise determination of time since death plays a major role in forensic routine. Currently available techniques for estimating the postmortem interval (PMI) are restricted to specific ...time periods or cannot be applied for individual case-specific reasons. During recent years, it has been repeatedly demonstrated that Western blot analysis of postmortem muscle protein degradation can substantially contribute to overcome these limitations in cases with different background. Enabling to delimit time points at which certain marker proteins undergo distinct degradation events, the method has become a reasonable new tool for PMI delimitation under various forensic scenarios. However, additional research is yet required to improve our understanding of protein decomposition and how it is affected by intrinsic and extrinsic factors.
Since there are temperature limits for proteolysis, and investigators are confronted with frozen corpses, investigation of the effects of freezing and thawing on postmortem protein decomposition in the muscle tissue is an important objective to firmly establish the new method. It is also important because freezing is often the only practical means to intermittently preserve tissue samples from both true cases and animal model research.
Methods
Sets of dismembered pig hind limbs, either freshly detached non-frozen, or thawed after 4 months of freeze-storage (
n
= 6 each), were left to decompose under controlled conditions at 30 °C for 7 days and 10 days, respectively. Samples of the M. biceps femoris were regularly collected at predefined time points. All samples were processed via SDS-PAGE and Western blotting to identify the degradation patterns of previously characterized muscle proteins.
Results
Western blots show that the proteins degrade predictably over time in precise patterns that are largely unaffected by the freeze-and-thaw process. Investigated proteins showed complete degradation of the native protein band, partly giving rise to degradation products present in distinct time phases of the decomposition process.
Conclusion
This study provides substantial new information from a porcine model to assess the degree of bias that freezing and thawing induces on postmortem degradation of skeletal muscle proteins. Results support that a freeze–thaw cycle with prolonged storage in frozen state has no significant impact on the decomposition behavior. This will help to equip the protein degradation–based method for PMI determination with a robust applicability in the normal forensic setting.
Human activities are changing landscape structure and function globally, affecting wildlife space use, and ultimately increasing human-wildlife conflicts and zoonotic disease spread. Capybaras ...(Hydrochoerus hydrochaeris) are linked to conflicts in human-modified landscapes (e.g. crop damage, vehicle collision), as well as the spread and amplification of Brazilian spotted fever (BSF), the most human-lethal tick-borne disease in the world. Even though it is essential to understand the link between capybaras, ticks and BSF, many knowledge gaps still exist regarding the effects of human disturbance in capybara space use. Here, we analyzed diurnal and nocturnal habitat selection strategies of capybaras across natural and human-modified landscapes using resource selection functions (RSF). Selection for forested habitats was higher across human-modified landscapes, mainly during day- periods, when compared to natural landscapes. Across natural landscapes, capybaras avoided forests during both day- and night periods. Water was consistently selected across both landscapes, during day- and nighttime. Distance to water was also the most important variable in predicting capybara habitat selection across natural landscapes. Capybaras showed slightly higher preferences for areas near grasses/shrubs across natural landscapes, and distance to grasses/shrubs was the most important variable in predicting capybara habitat selection across human-modified landscapes. Our results demonstrate human-driven variation in habitat selection strategies by capybaras. This behavioral adjustment across human-modified landscapes may be related to increases in A. sculptum density, ultimately affecting BSF.
Resin cement diffusion into dentin may differ as a function of the pre-treatment regimen. Since self-adhesive cements do not require substrate pre-treatment for luting, penetration of and interaction ...with the underlying dentin are questioned. We hypothesized that differences in the resin cement diffusion into dentin may exist among current commercial adhesive cements. Composite cylinders were luted on mid-coronal dentinal surfaces by an etch-and-rinse cement (Calibra), a self-etching system (Panavia F 2.0), and 4 self-adhesive cements (Multilink Sprint, Rely X Unicem, G-Cem, Bis-Cem). Dentin/cement interfacial characteristics were analyzed by a staining technique (Masson’s trichrome) and by scanning electron microscopy. Conventional acid etching resulted in partially infiltrated adhesive interfaces differing from those achieved with the application of self-etching primer. No hybrid layer and/or resin tag formation was detectable at the interfaces bonded with self-adhesive cements. Limited decalcification/infiltration was observed for self-adhesive cements into the underlying dentin. Self-adhesive cements were not able to demineralize/dissolve the smear layer completely.
The estimation of the postmortem interval (PMI) is of critical importance in forensic routine. The most frequently applied methods, however, are all restricted to specific time periods or must be ...excluded under certain circumstances. In the last years it has been shown that the analysis of muscle protein degradation has the potential to contribute to according delimitations in practice. In particular, upon biochemical analysis, the specific time points of degradation events provide reasonable markers for PMI delimitation. Nevertheless, considerable research is yet required to increase the understanding of protein decomposition and how it is affected by individual and environmental influencing factors. This is best investigated under standardized conditions, however, a considerate selection of proxies, regarding costs, effort, and expected outcome is required. Here, we use pigs to compare muscle protein decomposition in whole bodies and dismembered body parts (amputated hind limbs). Not only do experiments on body parts reduce the costs and allow easier handling in basic research, but also they aid to investigate the practical application of PMI estimation in dismembered body parts, or other extensive injuries, which are not unusual scenarios in crime investigation. Specifically, we investigated whether there are differences in the degradation rates of selected muscle proteins, sampled from dismembered legs and from hind limbs attached to whole pig bodies. Our results show distinct time-dependent degradation patterns of muscle proteins in a predictable manner regardless of sample origin. We are able to demonstrate that amputated hind limbs are suitable proxies for the analysis of muscle protein degradation, especially to investigate certain influencing factors and establish according standardized models.
Abstract Behavior is a useful trait for comparative studies that provide the comprehension of phylogenetic relationships among species. Here, we present a description of two spiny-rats species’ ...behavioral repertoire, Clyomys laticeps and Trinomys setosus (Rodentia: Echimyidae). The affiliative and agonistic behavioral patterns were sampled during a three-year study of captive populations of wild animals. Observational data were collected in two phases under different arrangements of individuals in groups. We also compare the behavioral traits of T. setosus and C. laticeps with the known behavioral patterns of Trinomys yonenagae. We add categories to the previous descriptions of T. setosus and a standard ethogram for C. laticeps. Trinomys setosus showed a visual and vocal display we called foot-trembling, which was not described in this form and function for other species studied until now. We discuss the differences in their sociality levels and similarities and differences among behavior patterns and repertoires.
Resumo O comportamento é uma característica útil para estudos comparativos que fornecem a compreensão das relações filogenéticas entre as espécies. Apresentamos aqui uma descrição do repertório comportamental de duas espécies de ratos-de-espinho Clyomys laticeps and Trinomys setosus (Rodentia: Echimyidae). Os padrões comportamentais afiliativos e agonísticos foram amostrados durante um estudo de três anos em populações de animais silvestres em cativeiro. Os dados foram coletados em duas fases sob diferentes arranjos de indivíduos em grupos sociais. Comparamos as características comportamentais de T. setosus e C. laticeps com as da espécie mais conhecida, T. yonenagae. Adicionamos categorias às descrições anteriores de T. setosus, e um etograma padrão para C. laticeps. Trinomys setosus mostrou uma exibição visual e vocal que chamamos de saltitar, que não foi descrito nesta forma e função para outras espécies do gênero estudado até agora. Discutimos diferenças nos níveis de socialidade e similaridades e diferenças entre os padrões comportamentais e repertórios.
The objective of this prospective clinical trial was to investigate the influence of the residual coronal structure of endodontically treated teeth and the type of cement used for luting fiber posts ...on four-year clinical survival. Two groups (n = 60) were defined, depending on the amount of residual coronal dentin after abutment build-up and final preparation: (1) more than 50% of coronal residual structure; and (2) equal to or less than 50% of coronal residual structure. Within each group, teeth were randomly divided into 2 subgroups (n = 30) according to the material used for luting fiber posts: (A) resin core build-up material, Gradia Core; or (B) self-adhesive universal cement GCem Automix. The rate of success was assessed based on clinical and intra-oral radiographic examinations at the follow-up after 6, 12, 24, 36, and 48 months. The highest 48-month success and survival rates were recorded in group 1A (90% and 100%, respectively), whereas teeth in group 2B exhibited the lowest performance (63.3% success rate, 86.6% survival rate). Cox regression analysis revealed that neither the amount of coronal residual structure nor the luting material significantly influenced the failure risk (p > .05) (ClinicalTrials.gov, NCT01532947).
Self-disclosures and self-interventions are generally considered useful, although their role remains controversial. This is mainly due to the heterogeneity of research findings on this topic, ...characterized by the lack of a clear definition of self-disclosure and of systematic and effective methods to evaluate the short- and long-term effects of such interventions. To clarify the concept of self-disclosure, it is essential to understand whether its definition comprehends different interventions, which may have different effects. An accurate and reliable evaluation of such effects may allow clinicians and researchers to differentiate among different kinds of self-disclosures. In this theoretical article, we propose a method to evaluate the clinical efficacy of self-disclosures interventions. This method is based upon the monitoring of motivational systems (Liotti, 2005), which allows therapists to assess the cooperative attunement between patient and therapist in real-time. Our central assumption is that the evaluation of the cooperative attunement between patient and therapist, as well as the patient’s metacognitive functioning, is primary to assess the use of clinical interventions. Indeed, these factors – either acting on their own or synergistically – are the mark, the essence, and the reflection of the therapeutic alliance, which is the strongest predictor of treatment’s outcome.
An accurate estimation of the postmortem interval (PMI) is a central aspect in forensic routine. Recently, a novel approach based on the analysis of postmortem muscle protein degradation has been ...proposed. However, a number of questions remain to be answered until sensible application of this method to a broad variety of forensic cases is possible. To evaluate whether altered in vivo protein metabolism interferes with postmortem degradation patterns, we conducted a comparative study. We developed a standardized animal degradation model in rats, and collected additional muscle samples from animals recovering from muscle injury and from rats with developed disuse muscle atrophy after induced spinal cord injury. All samples were analyzed by SDS-PAGE and Western blot, labeling well-characterized muscle proteins. Tropomyosin was found to be stable throughout the investigated PMI and no alterations were detected in regenerating and atrophic muscles. In contrast, significant predictable postmortem changes occurred in desmin and vinculin protein band patterns. While no significant deviations from native patterns were detected in at-death samples of disuse muscle atrophy, interestingly, samples of rats recovering from muscle injury revealed additional desmin and vinculin degradation bands that did not occur in this form in any of the examined postmortem samples regardless of PMI. It remains to be investigated whether in vivo-altered metabolism influences postmortem degradation kinetics or if such muscle samples undergo postmortem degradation in a regular fashion.
The delimitation of the postmortem interval (PMI) is of utmost importance in forensic science. It is especially difficult to determine the PMI in advanced decomposition stages and/or when dead bodies ...are found under uncommon circumstances, such as tents, or other (semi-) enclosed environments. In such cases, especially when insect access is restricted, morphological assessment of body decomposition is one of the remaining approaches for delimitation of the PMI. However, as this method allows only vague statements/indications about the PMI, it is required to develop new and more reliable methods. One of the most important candidates is the biochemical analysis of protein degradation. In this regard, it has been demonstrated that specific skeletal muscle protein degradation patterns characterize certain time points postmortem and thus can be used as markers for PMI estimation. In order to test this method in different micro-environments, a pilot study using ten pig carcasses was conducted in summer in Northern Germany. The cadavers were openly placed outside (freely accessible for insects), as well as enclosed in tents nearby, and left to decompose to investigate decomposition processes over a time course of 10 days. Muscle samples of the M. biceps femoris were collected on a regular basis and processed via SDS-PAGE and degradation patterns of selected proteins identified by Western blotting. In addition, morphological changes of the cadavers during decomposition were assessed using the total body score (TBS). Results showed that postmortem protein degradation patterns are largely consistent between treatment groups (open field versus tents) despite major morphological differences in the decomposition rate. This field study provides evidence that muscle protein degradation is mostly unaffected by different levels of exposure, making it a sufficient candidate for PMI delimitation under various circumstances.
Adhesive dentistry should effectively restore the peripheral seal of dentin after enamel removal. We hypothesize that non-rinsing, simplified, one-step self-etch adhesives are effective for ...minimizing dentin permeability after tooth preparation procedures. Crown preparations in vital human teeth were sealed with Adper Prompt, Xeno III, iBond, or One-Up Bond F. Epoxy resin replicas were produced from polyvinyl siloxane impressions for SEM examination. Dentin surfaces from extracted human teeth were bonded with these adhesives and connected to a fluid-transport model for permeability measurements and TEM examination. Dentinal fluid droplets were observed from adhesive surfaces in resin replicas of in vivo specimens. In vitro fluid conductance of dentin bonded with one-step self-etch adhesives was either similar to or greater than that of smear-layer-covered dentin. TEM revealed water trees within the adhesives that facilitate water movement across the polymerized, highly permeable adhesives. Both in vitro and in vivo results did not support the proposed hypothesis.
PDF
