Assessment of renal glomerulosclerosis and thickness of the carotid intima-media complex as a means of age estimation in Western European bodies

Introduction The estimation of age-at-death of unidentified cadavers is a central aspect of the identification process. With increasing age, the incidence of glomerulosclerosis and the thickness of the carotid wall have been observed to also increase. This correlation has been demonstrated in various international histological studies. The aim of our study was to assess whether these correlations also apply to a Western European population. Methodology In this retrospective observational study, kidney and common carotid artery samples from 216 cases autopsied at the Institute of Legal Medicine at the Justus-Liebig University in Giessen, Germany, were examined. Only cases with available tissue samples from both body sides were included. Exclusion criteria were poor sample quality and an age younger than 21 years. After histological processing, the tissue samples were assessed and digitally evaluated. Regression and classification analyses were used to investigate the correlation between age-at-death and intima-media thickness and age-at-death and the incidence of renal glomerular sclerosis. Results Of the 216 autopsy cases, 183 were included for evaluation. Analysis of the carotid artery segments showed a strong correlation (Pearson correlation coefficient r = 0.887) between the intima-media-complex thickness and chronological age. Classification of the glomerulosclerotic incidence showed a correlation of 37.7–43.1% with the predicted age group. Discussion Both the intima-media thickness and the proportion of sclerotic glomeruli can be used to estimate age in Western European cadavers. On the basis of these results, both methods are suited to supplement other already established methods for age-at-death estimation in the identification of an unknown cadaver.

Mediators involved in tooth resorption following delayed replantation: an experimental study

Inflammatory and replacement tooth resorption are common outcomes following tooth replantation, biological mediators involved in these processes are widely unknown. The aim study was to investigate molecules involved in tooth resorption following permanent tooth avulsion and delayed replantation. Dog premolars were extracted and kept dry for 20, 60 and 90 minutes (n= 30). The teeth were replanted, splinted. After 120 days, the animals were euthanized and tissues were removed for histological processing. Slides were stained for microscopic analysis, submitted to tartrate resistant acid phosphatase (TRAP) histoenzymology and immunostained for RANK, RANKL, OPG, alkaline phosphatase and periostin. Data obtained were submitted to statistical analysis using the chi-square, Fisher and one-way ANOVA tests (alpha= 0.05). In inflammatory resorption areas, TRAP + and RANK + osteoclasts surrounding the replanted teeth were identified, regardless of the extra-alveolar time. RANKL synthesis in this region was higher in longer extra-alveolar times and was more intense after keeping the tooth dry for 90 minutes compared to other periods. In the replacement reabsorption area, there was lower synthesis of periostin and higher alkaline phosphatase production. Inflammatory resorption was characterized by osteoclast recruitment and RANKL synthesis and replacement resorption was characterized by inhibition of periostin and alkaline phosphatase syntheses.

EXTRA VIRGIN OLIVE OIL EFFECT IN THE COMBINATION OF ALCOHOL AND BENZENE ON THE KIDNEY OF RATTUS NOVERGICUS

Objective: This study investigated the use of extra virgin olive oil (EVOO), which is said to possess anti-inflammatory components as an ameliorative substance for the induced inflammation. Methods: Forty-eight female Wistar rats were used for this study. They were divided into eight (1–8) groups (n=6); Control, Ethanol, Benzene, E+B, E+O, B+O, E+B+O with the administration of 25% Ethanol, 200 mg/kg/b.w Benzene, 2 ml (O) EVOO in respect to the tagged grouping names. Animals were euthanized through cervical dislocation after the last day of administration and the liver was excised and part was fixed in formalin solution of 10% for histological processing and the other part was homogenized for biochemical assay in phosphate buffer before centrifugation. Results: Histological evidence creates an understanding of the nephrons in which benzene and ethanol manifest likewise the mitigated ability of EVOO. The relative organ weight provides information on the extent of the damage on nephrons. The kidney test level is significantly expressed. Conclusion: Ethanol and benzene in combination cause serious damage; also, they separately induce nephrons dysfunction. EVOO was shown to mitigate these nephrons damages.

Histo Hepatic Alterations in Commercially Important Fish (Pseudoplatystoma corruscans Spix & Agassiz, 1829) Exposed to a Glyphosate-Based Herbicide

The presence of herbicides in the aquatic environment can cause different effects at all levels of biological organization. This study aimed to evaluate the hepatic alterations of the herbicide Roundup WG® on juvenile Pseudoplatystoma corruscans exposed to three different concentrations of this chemical compound: 0.25g/1000L, 0.50g/1000L, and 0.75g/1000L, plus control treatment (0.00g/1000L). The experiment lasted sixty days and, in the end, liver fragments were collected for further histological processing, using the hematoxylin-eosin (HE) technique. Only the control group showed a statistically significant increase in body mass and total length during the experiment. Blood glucose also showed no difference among the sample groups. The lesions found in the liver considered severe were hemorrhage, vacuolization, and hypertrophy of hepatocytes and the presence of free melanomacrophages, recorded in the groups exposed to 0.50 and 0.75g/1000L. The results obtained in the present study indicate that the herbicide Roundup WG® can promote liver alterations in Pseudoplatystoma corruscans.

Multifocal Urinary Tract Metastasis of Colorectal Carcinoma

<b><i>Introduction:</i></b> Secondary urinary tract tumors are uncommon findings and mainly evolve by direct invasion from adjacent organs. Actual metastatic involvement often develops in the urinary bladder, while the upper urinary tract is infrequently affected. In addition, the lungs, breast, and prostate gland are the usual primary sites. Colorectal carcinoma (CRC) may spread to the ureter directly or seeds via vascular or lymphatic channels. It may pose struggles in the differential diagnosis because CRC shares standard pathologic features with the primary adenocarcinoma of the urinary tract. <b><i>Case Presentation:</i></b> We describe the case of an 81-year-old man who was referred to our hospital with a distal ureteral tumor that was treated by a ureteronephrectomy. The histopathological and genetic analysis established the diagnosis of metastatic CRC along with 3 metastases in the renal pelvis. <b><i>Conclusion:</i></b> This rare case highlights the limitations of conventional histological processing, including immunohistochemistry, and it underlines the role of molecular investigations in certain circumstances.

Abstract P350: Myofibril Orientation In Cardiac Muscle And Its Implication For Heart Disease

Rationale: Myocyte disarray is a hallmark of cardiomyopathy. However, the orientation of individual myofibrils and myofilaments and how their alignment may be altered in disease progression have been largely underexplored. This oversight has been predominantly due to a paucity of methods for objective and quantitative analysis. Objective: To introduce a novel, less-biased approach to quantify myofibrillar and myofilament orientation in cardiac muscle under near physiological conditions and demonstrate its superiority versus traditional histological assessments. Methods and Results: Using small-angle X-ray diffraction, we first investigated changes in myofibrillar orientation at increasing sarcomere lengths in skinned, relaxed, wildtype mouse myocardium by assessing the angular spread of the 1,0 equatorial reflection (angle σ). At a sarcomere length (SL) of 1.9 μm, the angle σ was 0.23±0.01 rad, decreased to 0.19±0.01 rad at a SL of 2.1 μm, and further decreased to 0.15±0.01 rad at a SL of 2.3 μm (p<0.0001). Angle σ was significantly larger in R403Q (a MYH7 HCM model) porcine myocardium (0.24±0.01 rad) compared to WT myocardium (0.14±0.005 rad, p<0.0001) as well as in biopsied human heart failure tissue (0.19±0.006 rad) when compared to non-failing samples (0.17±0.007 rad, p=0.01). These data indicate that diseased myocardium suffers from myofibrillar disorientation compared to healthy controls. Finally, using control samples, we showed that traditional, histological-based analysis of disarray can be subject to user bias and/or sampling error and lead to false positives. Conclusions: Our method for assessing myofibrillar orientation limits the artifacts introduced by traditional histological processing and provides a precise and objective metric for phenotypically characterizing myocardium. The ability to obtain excellent X-ray diffraction patterns from frozen, biopsied human myocardium opens up new avenues of inquiry regarding the relation of myofibrillar structure to function in health and disease.

Tissue block staining and domestic adhesive tape yield qualified integral sections of adult mouse orbits and eyeballs

The standard histological processing procedure, which produces excellent staining of sections for most tissues, fails to yield satisfactory results in adult mouse orbits or eyeballs. Here, we show that a protocol using tissue block staining and domestic adhesive tapes resulted in qualified integral serial cryo-sections of whole orbits or eyeballs, and the fine structures were well preserved. The histological processing protocol comprises paraformaldehyde fixation, ethylenediaminetetraacetic acid decalcification, tissue block staining with hematoxylin and eosin, embedding, adhesive tape aided sectioning, and water-soluble mounting. This protocol was proved to be the best in comparison with seven other related existing histological traditional or non-traditional processing methods, according to the staining slice quality. We observed a hundred percent success rate in sectioning, collection, and mounting with this method. The reproducibility tested on qualified section success rates and slice quality scores confirmed that the technique is reliable. The feasibility of the method to detect target molecules in orbits was verified by successful trial tests on block immunostaining and adhesive tape-aided sectioning. Application of this protocol in joints, brains, and so on,—the challenging integral sectioning tissues, also generated high-quality histological staining sections.

Naringin Chelates Excessive Iron and Prevents the Formation of Amyloid-Beta Plaques in the Hippocampus of Iron-Overloaded Mice

Metal chelating agents are antioxidant agents, which decrease the reductive potential and stabilize the oxidized metal ion form. In this study, we evaluated the naringin capacity in chelating iron and preventing amyloid-beta plaque formation in the hippocampus of iron-overloaded mice. Thirty-five NMRI male mice (8–10 weeks old) were provided. The mice were classified into five groups. Iron dextran was administered as i.p. injection (100 mg/kg/day) four times a week for four subsequent weeks. The treated groups received 30 and 60 mg/kg/day naringin for a month. After histological processing, the brain sections were stained with Perls’ stain kit for iron spots, and Congo red was used to stain the brain and hippocampus for amyloid-beta plaques. 30 mg/kg/day of naringin was shown to decrease nonheme iron in an efficient manner; iron content in this group decreased to 16.83 ± 0.57 μg/g wet weight, a quantity as low as that observed in the normal saline-receiving group. The nonheme iron content in the mice receiving 60 mg/kg/day of naringin was 20.73 ± 0.65 μg/g wet weight. In addition, Aβ plaque numbers in CA1, CA3, and DG areas of the hippocampus decreased significantly following treatment with 30 or 60 mg/kg/day naringin. Naringin has a strong iron chelation capacity and is able to reduce the formation of amyloid plaques. So it can be useful for neuroprotection and prevention of Alzheimer’s disease.