Artificial intelligence assisted compositional analyses of human abdominal aortic aneurysms ex vivo.

Quantification of histological information from excised human abdominal aortic aneurysm (AAA) specimens may provide essential information on the degree of infiltration of inflammatory cells in different regions of the AAA. Such information will support mechanistic insight in AAA pathology and can be linked to clinical measures for further development of AAA treatment regimens. We hypothesize that artificial intelligence can support high throughput analyses of histological sections of excised human AAA. We present an analysis framework based on supervised machine learning. We used TensorFlow and QuPath to determine the overall architecture of the AAA: thrombus, arterial wall, and adventitial loose connective tissue. Within the wall and adventitial zones, the content of collagen, elastin, and specific inflammatory cells was quantified. A deep neural network (DNN) was trained on manually annotated, Weigert stained, tissue sections (14 patients) and validated on images from two other patients. Finally, we applied the method on 95 new patient samples. The DNN was able to segment the sections according to the overall wall architecture with Jaccard coefficients after 65 epocs of 92% for the training and 88% for the validation data set, respectively. Precision and recall both reached 92%. The zone areas were highly variable between patients, as were the outputs on total cell count and elastin/collagen fiber content. The number of specific cells or stained area per zone was deterministically determined. However, combining the masks based on the Weigert stainings, with images of immunostained serial sections requires addition of landmark recognition to the analysis path. The combination of digital pathology, the DNN we developed, and landmark registration will provide a strong tool for future analyses of the histology of excised human AAA. In combination with biomechanical testing and microstructurally motivated mathematical models of AAA remodeling, the method has the potential to be a strong tool to provide mechanistic insight in the disease. In combination with each patients' demographic and clinical profile, the method can be an interesting tool to in supportof a better treatment regime for the patients.

Vitellogenin as biomarker for estrogenicity in flounder Platichthys flesus in the field and exposed to 17α-ethinylestradiol via food and water in the laboratory.

The ability of 17α-ethinylestradiol (EE2) to elevate vitellogenin levels were investigated in male flounder Platichthys flesus and vitellogenin concentrations in flounders from the Danish coastal environment were determined. Male flounders were exposed to 17α-ethinylestradiol (EE2) via food or water. Average vitellogenin concentrations in the control fish ranged between 25 and 100 ng mL(-)(1). Exposure to 5.1, 8.1 and 16.8 ng EE2 L(-)(1) in water and 500 and 5000 ng EE2 kg(-)(1) body weight (bw) every second day in the food increased the plasma vitellogenin concentration in a concentration and time dependent manner, whereas exposure to 2.7 ng EE2 L(-)(1) in water for 21 d and 5 and 50 ng EE2 kg(-)(1) bw for 12 days in the food did not. EE2 could be detected in liver and testes (but not in muscle) after exposure to 8.1 and 16.8 ng EE2 L(-)(1) in the water and 5000 ng EE2 kg(-)(1) bw in the food; the highest concentration was 6 ng g(-)(1) wet weight in liver. The majority of the male flounders collected from nine coastal Danish sites from 1999 to 2004 had vitellogenin concentrations below 100 ng mL(-)(1), and only at two sites moderate estrogenic inputs were indicated.

Sex hormone concentrations and gonad histology in brown trout (Salmo trutta) exposed to 17beta-estradiol and bisphenol A.

The impact of 17beta-estradiol (E2) and bisphenol A (BPA) on steroid hormone levels and gonad development in brown trout (Salmo trutta) was determined. Exposure took place from 0 to 63 days post-fertilisation (dpf) and gonad development was followed till 400 dpf. The onset of xenoestrogen metabolism was examined by measurements of whole body concentrations of bisphenol A (BPA) and its conjugation product bisphenol A glucuronic acid (BPAGA). Exposure to 500 ng E2/l led to an increase in E2 levels in the embryos and fry while 10 ng E2/l did not. Metabolic conversion of BPA to BPAGA began during the first weeks of embryonic development. Few consistent effects were found on the sex differentiation of the brown trout. Only one intersex fish (4.5%) was found among male fish at 400 dpf exposed to 500 ng E2/l. Females with male germ cells among the normally developing oocytes were observed in all groups (in up to 50% of the female fish, independently of exposure regime). The fact that exposure to 500 ng E2/l only caused subtle effects in a small number of individuals indicates that exposure during early life stages results in little to no induction of endocrine disruption in brown trout.

Gonad histology and vitellogenin concentrations in brown trout (Salmo trutta) from Danish streams impacted by sewage effluent.

Brown trout (Salmo trutta) collected from a number of Danish streams impacted by sewage effluent were examined for alterations to gonadal development and induction of vitellogenin synthesis. Among fish collected in June/July 2000/2001 and November 2002 higher levels of plasma vitellogenin were found in males from six streams impacted by sewage effluent compared to males from their respective reference sites. A direct non-competitive ELISA was developed for brown trout vitellogenin in order to perform the vitellogenin measurements. Intersex in females with no apparent relation to sewage effluent exposure was observed at all sites. In one stream, male brown trout with a very high level of vitellogenin were concomitantly found to have a high degree of vacuolation of the testes and a presence of only the early spermatogenic stage, spermatogonia. The cause of these alterations to the testis structure is unclear. However, as a high level of plasma vitellogenin in these males indicated estrogenic exposure, the vacuolation might also be a result of endocrine disruption causing delayed or disrupted spermatogenesis.

Intersex in wild roach (Rutilus rutilus) from Danish sewage effluent-receiving streams.

Roach (Rutilus rutilus) from Danish streams that receive discharges of domestic sewage effluent were examined for the presence of alterations to gonadal development. In male roach, intersex was observed at a prevalence of 4.5-5% at reference sites and 6.7-6.5% at sewage effluent-impacted sites. A positive correlation was found between sewage effluent load and intersex frequency among male roach. The highest frequency of intersex (26.5%) was found in the stream Kristrup Landkanal, which had the highest percentage and load of sewage effluent (87,578 population equivalents). Further, a tendency to an average higher severity of the phenomenon (calculated as an intersex index value) was seen in males from streams impacted by sewage effluent compared to males from reference sites. However, this was significant only in male fish from Egaa Brook. Among roach from the Kristrup Landkanal 5.8% of male and 0.8% of female roach were infected with the parasite Pleistophora mirandellae, causing degenerative changes in testes and ovaries. No correlation was seen between the intersex condition and the presence of P. mirandellae in the gonads of roach.