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Abstract

Background

The hemostatic system cooperates with proteolytic degradation in processes allowing abdominal aortic aneurysm (AAA) formation. In previous studies, it has been suggested that aneurysm rupture depends on intraluminal thrombus (ILT) thickness, which varies across each individual aneurysm. We hypothesized that hemostatic components differentially accumulate in AAA tissue in relation to ILT thickness. Thick (A1) and thin (B1) segments of ILTs and aneurysm wall sections A (adjacent to A1) and B (adjacent to B1) from one aneurysm sac were taken from 35 patients undergoing elective repair.

Methods

Factor levels were measured using enzyme-linked immunosorbent assay of protein extract.

Results

Tissue factor (TF) activities were significantly higher in thinner segments of AAA (B1 vs A1, P = .003; B vs A, P < .001; B vs A1, P < .001; B vs B1, P = .001). Significantly higher tissue plasminogen activator was found in thick thrombus-covered wall segments (A) than in B, A1, and B1 (P = .015, P < .001, and P < .001, respectively). Plasminogen concentrations were highest in ILT. Concentrations of α2-antiplasmin in thin ILT adjacent walls (B) were higher compared with wall (A) adjacent to thick ILT (P = .021) and thick ILT (A1; P < .001). Significant correlations between levels of different factors were mostly found in thick ILT (A1). However, no correlations were found at B sites, except for a correlation between plasmin and TF activities (r = 0.55; P = .004).

Conclusions

These results suggest that higher TF activities are present in thinner AAA regions. These parameters and local fibrinolysis may be part of the processes leading to destruction of the aneurysm wall.

Abstract

Aims

Toll-like receptors (TLR) and apoptosis were indicated as important factors in heart failure. Our aim was to characterize the morphological pattern of apoptosis, TLR2, TLR4, and TLR6 expression in female rat hearts in the model of takotsubo syndrome (TTS).

Main methods

60 Sprague-Dawley female rats were treated with a single dose of 150 mg/kg b.wt. of isoprenaline (ISO) or 0.9% NaCl (controls). Hearts were collected 24, 48, 72?h and 7?days post-ISO injection. 32/60 hearts were used in immunohistopathological studies and 28/60 in real time.

Key findings

Apoptosis was observed 24h post-ISO in cardiomyocytes, 24, 48, 72?h and 7 days post-ISO in infiltrating inflammatory cells, 7?days post-ISO in endothelial cells of vessels. Diffuse TLR4CD68 (CD68, a macrophage marker) and TLR6CD68 positive cells and TLR2, TLR4, TLR6 mononuclear cells were observed in both acute and recovery phase of TTS. In the foci located in the neighborhood of damaged (necrotic/apoptotic) cardiomyocytes in TTS, high (strong) protein expression of TLR2 (TLR2high) was observed: 24, 48, 72?h post-ISO; TLR4high-48 and 72 h post-ISO; TLR6high-48 h post-ISO. Whereas in cardiomyocytes of remote myocardium: TLR2high -72?h post-ISO; TLR4high-24 and 72?h post-ISO; TLR6high-24?h post-ISO. TLR2 mRNA was down-regulated 48 and 72?h post-ISO whereas TLR4 up-regulated 7?days post-ISO.

Significance

The expression pattern of apoptosis and TLR differs in the course of TTS in comparison with the control rats. We hypothesize that innate immunity and apoptosis may play a crucial role in TTS pathophysiology.

Abstract

Very little is known about the mechanisms by which malignant ascites modulates the cancer-promoting activity of human peritoneal mesothelial cells (HPMCs). Because malignant ascites induces pro-tumoral senescence in HPMCs, here we examined if this effect could be driven by oxidative stress. The study showed that malignant ascites generated by serous ovarian tumors induced oxidative damage to the DNA (gamma H2A.X, 53BP1, 8-hydroxy-2-deoxyguanosine) and lipids (8-isoprostane) in HPMCs as well as increased the production of mitochondrial superoxides and cellular peroxides in these cells. This activity coincided with increased activity of two enzymes involved in the mitochondrial production of oxidants, i.e. cytochrome c oxidase and NADH dehydrogenase, decreased mitochondrial inner membrane potential, increased mitochondrial mass, and increased the activity of peroxisome proliferator-activated receptor gamma coactivator-1 alpha. Increased production of superoxides and peroxides in cells subjected to the malignant ascites was effectively reduced when the fluid was pre-incubated with neutralizing antibodies against hepatocyte growth factor. Moreover, when HPMCs subjected to the malignant ascites were protected against oxidative stress with a spin-trap scavenger of reactive oxygen species, they displayed decreased expression of senescence-associated beta-galactosidase and their potential to stimulate cancer cell adhesion, proliferation, and migration was significantly diminished. Collectively, our findings indicate that improved ovarian cancer cell progression in response to HPMCs exposed to malignant ascites may be associated with the development of profound oxidative stress in these cells.

Abstract

We examined the direct effects of toxaphene and endrin, chlorinated insecticides that are widespread in the environment, on myometrial contractions and on the secretion of hormones involved in regulating these contractions. Granulosa, luteal, endometrial and myometrial cells, and myometrial strips from non-pregnant cows were incubated with both insecticides at environmentally relevant doses.

Toxaphene inhibited and endrin stimulated the secretion of testosterone and oestradiol from granulosa cells. Toxaphene also inhibited and endrin stimulated the expression of the mRNA encoding the precursor of oxytocin (OT), as well OT secretion in luteal cell cultures. Moreover, endrin increased OT secretion from granulosa cells. Neither insecticide exerted an effect on progesterone secretion from luteal cells. Only toxaphene decreased the secretion of prostaglandins (PGF2 and PGE2) from endometrial cells. Meanwhile, only endrin decreased basal myometrial contractions, which was accompanied by inhibition of PGF2 secretion from the myometrium. Both endrin and toxaphene also decreased the force of the OT-stimulated myometrial contractions, whereas only toxaphene inhibited the stimulatory effect of OT on the force of myometrial contractions.

In contrast to endrin, toxaphene decreased synthesis and secretion of one of the primary stimulators of myometrial contractions (OT) and indirectly inhibited OT signal reception in the myometrium by reducing E2 secretion. Both insecticides decreased OT-stimulated myometrial contractions; therefore, they may inhibit further transmission of the OT signal. Moreover, endrin inhibited basal myometrial contractions, potentially resulting from reduced PGF2 secretion from the myometrium. Our data indicate the potential of these insecticides to disturb the course of the oestrous cycle or fertilisation.

VIP changes during daytime in chicken intrinsic choroidal neurons

Posted by B Hohberger, C Jessberger, M Zenkel, et al. on 2018-03-21 20:10:21

Abstract

Purpose

Ocular autonomic control is mediated by sympathetic and parasympathetic nerve fibres. Their interactions are complemented by primary afferent nerve fibers of and intrinsic choroidal neurons (ICN). As the vasodilatative neuropeptide, vasoactive intestinal peptide (VIP), is expressed in extrinsic and intrinsic ocular neurons, it is of special interest in ophthalmic research. Since circadian changes of ocular blood flow are known in humans and birds, this study aimed at investigating VIP expression at different daytimes in chicken choroid, the preferred model species in ICN research.

Methods

12 eyes of 12 chickens were retrieved, slaughtered at 8.00-9.30 a.m. (n=6) and 8.00 p.m. (n=6), respectively, and choroidal wholemounts were prepared for immunofluorescence of VIP. VIP-positive ICN of both groups were quantified and density of VIP-positive axons assessed semi-quantitatively. In 28 additional eyes retrieved in the morning (n=14) and evening (n=14), choroidal VIP content was determined by ELISA. Morning and evening data were analyzed statistically.

Results

(1) Numbers of VIP positive neurons differed significantly between morning: (239.17±113.9) and evening: (550.83±245.7; p=0.018). (2) Numbers of VIP-positive perikarya were significantly more accumulated in the temporal part of the choroid in the evening than in the morning (p=0.026). (3) VIP positive axon density was found to be similar throughout the choroid in the morning and evening. (4) ELISA demonstrated a significant difference of VIP content (p=0.012) in tissues harvested in the morning (145.41±43.3?pg/ml) compared to evening (221.44±106.3?pg/ml).

Conclusions

As VIP positive axon density was similar in the morning and the evening throughout the choroid, PPG and ICN seemed to contribute equally to the axon network. Yet, changes in the total choroidal VIP content, the numbers of VIP positive perikarya, reflecting the intracellular VIP content, and their topographical distribution at two different days-times argue for a different status of activation of both neuronal sources. The higher VIP content in the evening, compared to the morning, correlates with a known circadian rhythm of a lower IOP and a higher choroidal thickness at night. Thus, these changes may argue for a potential role of ICN in the regulation of ocular homeostasis and integrity.

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