Browsing by Subject "Forensic pathology"
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- PublicationOpen AccessCaracterización del pulmón como órgano diana de la muerte por sumersión: evaluación del estrés oxidativo y concentraciones de elementos traza(Universidad de Murcia, 2022-09-28) Barrera Pérez, Estefanía; María Dolores Pérez Cárceles; Agustín Sibón Olano; Escuela Internacional de DoctoradoEl diagnóstico de asfixia por sumersión sigue siendo una de las muertes más difíciles y controvertidas de establecer. Representa una de las principales causas de muerte no intencional a nivel mundial. En la mayoría de las ocasiones, el patólogo forense precisa para llegar a un diagnóstico certero, no solo de los hallazgos de autopsia, sino de una serie de pruebas complementarias tales como estudios histopatológicos, biológicos y químicos. A pesar de ser el pulmón el órgano más afectado en la muerte por sumersión, existe poca información sobre el impacto que tiene en este órgano este tipo de muerte. Los objetivos de nuestro trabajo fue investigar la concentración de elementos traza en los pulmones de sujetos que habían fallecido por diferentes causas de muerte para evaluar la capacidad de discriminación de los elementos traza en el diagnóstico de la muerte por sumersión en agua salada. Y conocer los procesos de estrés oxidativo en las muertes por sumersión en agua salada y otras causas de muerte, analizando los niveles de marcadores de estrés oxidativo (MDA y GSH) en lóbulos pulmonares, así como su correlación con alteraciones en la expresión inmunohistoquímica de la proteína A del surfactante pulmonar. Material y métodos: se analizaron un total de 11 elementos traza en 74 casos forenses mediante Espectrometría de Masas con Plasma Acoplado Inductivamente (ICP-MS). Para identificar alteraciones ultraestructurales y determinar los niveles de elementos traza en las muestras de pulmón se utilizó microscopía electrónica de barrido asociado a espectroscopia de rayos X de dispersión de energía. Por otra parte, estudiamos en una serie de 93 casos de autopsias forenses, los marcadores de estrés oxidativo mediante determinación colorimétrica de Malondialdehído (MDA), por espectrofotometría el GSH (método de Tiztze). Por último, para evaluar la expresión inmunohistoquímica de la PS-A se realizaron análisis histológicos utilizando el anticuerpo monoclonal de ratón anti-SPA humano PE-10. Resultados: se obtuvieron concentraciones significativamente más altas de Br y Sr, y concentraciones significativamente más bajas de Cu, Zn, Se, Cd y Pb en los casos de muerte por sumersión en agua salada frente a otras causas de muerte. Estos resultados fueron confirmados por PCA, que reveló una amplia separación entre las muertes por sumersión y el resto de las causas de muerte. Los niveles de MDA fueron superiores en ambos pulmones en los casos de muerte por sumersión en agua de mar en comparación con otras causas de muerte, existiendo diferencias estadísticamente significativas en las concentraciones de MDA entre los tres grupos de causas de muerte analizados. Del mismo modo, se observó niveles significativamente superiores de GSH en los casos de sumersión frente al resto de muertes, resultando ser más significativo en el pulmón derecho. Se obtuvo mayor expresión inmunohistoquímica de la SP-A en los casos de sumersión en agua salada que en las otras causas de muerte, siendo los niveles superiores en ambos pulmones. En el análisis de correlación entre los niveles de estrés oxidativo (MDA y GSH) en tejido pulmonar y el nivel de expresión de la SP-A se obtuvieron resultados positivos y significativos para los casos de sumersión en agua de mar, tanto en membrana alveolar como en el espacio alveolar. Conclusiones: la determinación de los elementos traza Br y Sr en pulmón junto con la determinación de los niveles de MDA y GSH en tejido pulmonar, así como el nivel de expresión de la SP-A puede ser de gran importancia en el diagnóstico de la muerte por sumersión en agua salada y circunstancias de la muerte. Un mejor conocimiento de la fisiología de la sumersión es importante por sus posibles repercusiones en la adopción de medidas en el abordaje de pacientes que han sobrevivido a un proceso de sumersión y también crucial en patología forense para una interpretación más correcta de los acontecimientos que conducen a la muerte por sumersión.
- PublicationOpen AccessCombining oxidative stress markers and expression of surfactant Protein A in lungs in the diagnosis of deawater drowning(MDPI, 2023-01-05) Legaz Pérez, Isabel; Barrera Pérez, Estefanía; Sibón, Agustín; Martínez Díaz, F.; Pérez Cárceles, María D.; Ciencias SociosanitariasBackground and Objectives. The diagnosis of seawater drowning (SWD) remains one of the most complex and contentious. It is one of the leading causes of unintentional death around the world. In most cases, the forensic pathologist must reach an accurate diagnosis from the autopsy findings and a series of complementary tests such as histopathological, biological, and chemical studies. Despite the lung being the most affected organ in death by submersion, there are few studies on this type of death’s impact on this organ. The aim was to investigate human lung cadavers of forensic cases due to different causes of death, the concentration of the oxidative stress markers malondialdehyde (MDA) and γ-glutamyl-l-cysteinyl glycine (GSH), and the relationship with the expression of surfactant protein A (SP-A) to try to discriminate SWD from other types of causes of death. Materials and Methods. A total of 93 forensic autopsy cases were analyzed. Deaths were classified into three major groups based on the scene, cause of death, and autopsy findings (external foam, frothy fluid in airways, overlapping medial edges of the lungs): (a) drowning in seawater (n = 35), (b) other asphyxia (n = 33), such as hangings (n = 23), suffocations (n = 6), and strangulation (n = 4), and (c) other causes (n = 25), such as multiple suffocations. Oxidative stress markers (MDA and GSH) and the immunohistochemical expression of SP-A were determined in both lungs. Results. MDA levels were statistically higher in both lungs in cases of SWD than in other causes of death (p = 0.023). Similarly, significantly higher levels of GSH were observed in SWD compared to the rest of the deaths (p = 0.002), which was more significant in the right lung. Higher immunohistochemical expression of SP-A was obtained in the cases of SWD than in the other causes of death, with higher levels in both lungs. The correlation analysis between the levels of oxidative stress (MDA and GSH) in the lung tissue and the expression level of SP-A showed positive and significant results in SWD, both in the alveolar membrane and the alveolar space. Conclusions. Determining the levels of MDA and GSH in lung tissue and the expression level of SP-A can be of great importance in diagnosing SWD and the circumstances of death. A better understanding of the physiology of submersion is essential for its possible repercussions in adopting measures in the approach to patients who have survived a submersion process. It is also necessary for forensic pathology to correctly interpret the events that lead to submersion.
- PublicationOpen AccessEditorial: skin lesion vitality assessment for forensic science: Current research and new perspectives(Frontiers Media, 2022-08-15) Legaz Pérez, Isabel; Fineschi, Vittorio; Madea, Burkhard; Bacci, Stefano; Ciencias Sociosanitarias
- PublicationOpen AccessEvaluating the link between BAFF system gene expression and acute rejection development in kidney transplantation(MDPI, 2022-07-07) Alfaro, Rafael; Llorente, Santiago; Jiménez Coll, Víctor; Martínez Banaclocha, Helios; Galián, José Antonio; Botella, Carmen; Moya Quiles, María Rosa; Muro Pérez, Manuel; Peña Moral, Jesús de la; Minguela, Alfredo; Legaz Pérez, Isabel; Muro, Manuel; Ciencias SociosanitariasB-cell activating factor (BAFF) system signaling is critical for B-cell homeostasis, effector functions, and tolerance maintenance in transplants, but it has not been studied in kidney transplant recipients (KTRs). The aim was to analyze the changes in BAFF system expression in KTRs with/without acute rejection (AR/NAR). The BAFF system expression was analyzed by qPCR in 40 KTRs. A meta-analysis of BAFF system expression and histological renal damage was identified by the Chronic Allograft Damage Index (CADI) and performed from the GEO database. Proliferation-inducing ligand (APRIL) expression increased at three- and six-months post-KT (p = 0.014 and p < 0.001). B-cell maturation antigen (BCMA) expression increased at six-months post-KT (p = 0.038). BAFF expression remained stable in NAR-KTRs, but was increased in CADI concerning the No-CADI group at one year (p = 0.008). BCMA expression increased in the CADI group at one- (p = 0.001) and six-years post-KT (p = 0.024). At three months, the transmembrane activator and calcium modulator interactor (TACI) gene significantly elevated KTRs with DSAs (donor-specific antibody; p = 0.034). KTRs with DSAs significantly increase the B-cell activating factor receptor (R-BAFF; p = 0.021) and TACI (p = 0.018) between pre- and three-month post-KT. Changes in the expression of the BAFF system increase during post-KTR in the development of AR and chronic allograft damage, and could be an important pathological tool to detect and prevent kidney graft outcomes.
- PublicationOpen AccessHistological findings and immunohistochemical surfactant protein A -SP-A- expression in asphyxia, Its application in the diagnosis of drowning(Murcia : F. Hernández, 2008) Pérez Cárceles, María Dolores; Sibón, A.; Vizcaya, M.A.; Osuna Carrillo-Albornoz, Eduardo Javier; Gómez Zapata., Maximiliano; Luna, A.; Martínez Díaz, F.The histopathological alterations that permit the diagnosis of death by asphyxia are very unspecific, although pulmonary alterations are of great importance in this respect. The postmortem diagnosis of drowning, particularly, continues to be one of the most difficult in forensic pathology. The aim of this study is to jointly evaluate microscopic findings and immunohistochemical surfactant protein A (SP-A) expression in the upper and lower lobes of lungs in different causes of death, and their possible application to the diagnosis of drowning. We studied 120 cadavers from subjects with a mean age of 48.73 years (SD 19.45; range 2-86 years), and with a mean post-mortem interval of 30 hours (SD 39.59; range 3-216 hours). According to the scene, cause and circumstances of death, and autopsy findings, cases were classified into groups as follows: (a) drowning (n=47); (b) other asphyxia (n=44) and (c) other causes (n=29). In the upper and lower lobes of lungs, histological studies of H&E staining and immunohistochemical surfactant protein A expression were made. The presence and severity of congestion, haemorrhage and oedema, together with immunohistochemical SP-A expression, may have a diagnostic value in differentiating asphyxia and drowning from other causes of death, and drowning from other types of asphyxia. Our findings suggest that both lobes should be investigated to establish the diagnosis, although the findings in the upper lobe might be the most important for differentiating the exact cause of death.
- PublicationOpen AccessImmunohistochemistry of connexin43 and zonula occludens-1 in the myocardium as markers of early ischemia in autopsy material(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2014) Kawamoto, Osamu; Michiue, Tomomi; Ishikawa, Takaki; Maeda, HitoshiImmunohistochemistry of the terminal complement complex (C5b-9) and fibronectin (FN) is useful to detect myocardial ischemia preceding necrosis in the postmortem diagnosis of sudden cardiac death. The present immunohistochemical study examined connexin43 (Cx43) and zonula occludens-1 (ZO-1) as markers of early myocardial ischemia in addition to the above-mentioned markers, using forensic autopsy cases of acute deaths due to myocardial infarction (MI, n=15) and acute ischemic heart disease (AIHD) without apparent myocardial necrosis (n=8), compared with those of acute mechanical asphyxiation (As, n=24) and drowning (D, n=10) as controls. Immunopositivities of each marker in the myocardium were semi-quantitatively graded by scoring. ZO-1, C5b-9 and FN were detected in the myocardial cytoplasm, whereas Cx43 and nonphosphorylated (np) Cx43 showed varied localizations at the intercalated disc, in the cytoplasm and along the lateral cell border. ZO-1 and FN showed a tendency to be detected more intensely in MI and IHD than in As and D. C5b-9 showed specific staining at the site of ischemia in MI (n=10/15) and AIHD (n=6/8), while the distribution of npCx43 was different in most cases of MI (n=14/15) and AIHD (n=5/8), compared with As and D; npCx43 positivity score was higher in the cytoplasm than at the intercalated disc, indicating redistribution due to myocardial ischemia. Such findings were detected in a few cases of As (n=3/24). These findings suggest that the combination of npCx43 and C5b-9 immunohistochemistry is useful for detecting early lesions of myocardial ischemia in sudden cardiac death.
- PublicationOpen AccessMicroRNA expression changes in kidney transplant: diagnostic efficacy of miR-150-5p as potential rejection biomarker, pilot study(MDPI, 2021-06-22) Alfaro, Rafael; Legaz Pérez, Isabel; Jiménez Coll, Víctor; El kaaoui El band, Jaouad; Martínez Banaclocha, Helios; Galián, José Antonio; Parrado, Antonio; Mrowiec, Anna; Botella, Carmen; Moya Quiles, María Rosa; Boix, Francisco; Peña Moral, Jesús de la; Minguela, Alfredo; Llorente, Santiago; Muro, Manuel; Ciencias SociosanitariasBackground: The kidney allograft biopsy is considered the gold standard for rejection diagnosis but is invasive and could be indeterminate. Several publications point to the role of miRNA expression in suggesting its involvement in the acceptance or rejection of organ transplantation. This study aimed to analyze microRNAs involved in the differentiation and activation of B and T lymphocytes from kidney transplant (KT) patients’ peripheral blood leukocytes to be used as biomarkers of acute renal rejection (AR). Methods: A total of 15 KT patients with and without acute rejection (AR/NAR) were analyzed and quantified by miRNA PCR array. A total of 84 miRNAs related to lymphocyte differentiation and activation B and T were studied. The functions and biological pathways were analyzed to predict the potential targets of differential expressed miRNAs. Results: Six miRNA were increased in the AR group (miR-191-5p, miR-223-3p, miR-346, miR-423-5p, miR-574-3p, and miR-181d) and miR-150-5p was increased in the NAR group. In silico studies showed a total of 2603 target genes for the increased miRNAs in AR, while for the decrease miRNA, a total of 1107 target-potential genes were found. Conclusions: Our results show that KT with AR shows a decrease in miR-150-5p expression compared to NAR, suggesting that the decrease in miR-150-5p could be related to an increased MBD6 whose deregulation could have clinical consequences.
- PublicationOpen AccessMonitoring of B cell in kidney transplantation: development of a novel clusters analysis and role of transitional B cells in transplant outcome(MDPI, 2021-04-01) Alfaro, Rafael; Legaz Pérez, Isabel; González Martínez, Gema; Jiménez Coll, Víctor; Martínez Banaclocha, Helios; Galián, José Antonio; Botella, Carmen; Peña Moral, Jesús de la; Moya Quiles, María Rosa; Campillo, José Antonio; Minguela, Alfredo; Llorente, Santiago; Muro, Manuel; Ciencias SociosanitariasBackground: B lymphocytes (BL) seem to play an important role in transplantation, although the and role of different subpopulations in monitoring and outcome is not clear. Our aim was to monitoring immunological profiles based on BL subpopulations in kidney recipients (KR) with the risk of acute rejection (AR). Methods: Monitoring of BL subpopulations was performed by flow cytometry in PBLs before transplantation and three and six months after transplantation (PTX). We used two methodological approaches, a traditional analysis, and a novel cluster analysis, to determine the association between BL subpopulations, AR incidence, and graft function. Results: After three months of PTX, KRs with a B phenotype enriched in transitional BL and plasmablasts had better kidney function and lower AR incidence. KRs with decreased transitional BL and plasmablasts were associated with lower kidney function and higher AR PTX. KRs that had an increase in transitional BL PTX had a better clinical outcome. The increase in transitory BL during PTX was also associated with an increase in Tregs. Indeed, KRs receiving thymoglobulin as induction therapy showed a slight decrease in the relative frequency of naive BLs after three months of PTX. Conclusion: The monitoring of BL subpopulations may serve as a non-invasive tool to improve immunological follow-up of patients after kidney transplantation. However, further studies are needed to confirm the obtained results, define cut-off values, and standardize more optimal and even custom/customized protocols.