Histopathology Image Analysis in Two Long‐Term Animal Experiments with Helical Flow Total Artificial Heart

Histopathological analysis can provide important information in long‐term experiments with total artificial heart (TAH). Recently, a new type of blood pump, the helical flow total artificial heart (HF‐TAH) was developed. This study aimed to investigate the changes in selected vital organs in animal experiments with implanted HF‐TAH. Samples from lung, liver, and kidneys from two female goats (No. 1301 and No. 1304) with implanted HF‐TAH were analyzed. Tissue samples were fixed in 10% formaldehyde and 4 µm thick transverse sections were stained with hematoxylin‐eosin (HE). Additional staining was done for detection of connective tissue (Masson‐Goldner stain) and for detection of iron (hemosiderin) deposits (Perls stain). Sections were scanned at 100× and 500× magnification with a light microscope. Experiment no. 1301 survived 100 days (cause of termination was heavy damage of the right pump); experimental goat no.1304 survived 68 days and was sacrificed due to severe right hydrodynamic bearing malfunction. Histopathological analysis of liver samples proved signs of chronic venostasis with limited focal necrotic zones. Dilated tubules, proteinaceous material in tubular lumen, and hemosiderin deposits were detected in kidney samples. Contamination of the organs by embolized micro‐particles was suspected at the autopsy after discovery of visible damage (scratches) of the pump impeller surface (made from titanium alloy) in both experiments. Sporadic deposits of foreign micro‐particles (presumably titanium) were observed in most of the analyzed parenchymal organs. However, the described deposits were not in direct connection with inflammatory reactions in the analyzed tissues. Histopathological analysis showed the presence of minimal contamination of the lung, kidney, and liver tissue samples by foreign material (titanium very likely). The analysis showed only limited pathological changes, especially in liver and kidneys, which might be attributed to the influence of artificial perfusion often observed in chronic TAH experiments.     

Catecholamines and their <Emphasis Type="Italic">O</Emphasis>-methylated metabolites in vitreous humor in hypothermia cases

Purpose

The aim of this study was to assess the diagnostic value of catecholamines and their O-methylated metabolites in vitreous humor samples in identifying antemortem cold exposure and fatal hypothermia in the forensic casework.

Methods

A total of 80 autopsy cases (40 hypothermia fatalities and 40 cases in which hypothermia as the main or contributory cause of death was excluded) were selected for this study. Catecholamines and their O-methylated metabolites were measured in urine and vitreous humor samples collected at autopsy.

Results

Urine catecholamine and their O-methylated metabolite concentrations were significantly higher in hypothermia-related deaths. On the other hand, measurements in vitreous humor samples did not reveal statistically significant differences between hypothermia-related deaths and controls.

Conclusions

Globally considered, our findings seem to suggest that, contrary to urine catecholamines and their O-methylated metabolites, vitreous levels of these compounds appear to be of limited value in characterizing human antemortem stress reactions due to cold exposure and can hardly be used in the forensic setting to support the diagnosis of hypothermia.

     

fMRI evaluation of hemispheric language dominance using various methods of laterality index calculation

Several functional MR imaging studies evaluating the lateralisation of linguistic functions in patients who underwent Wada testing have been reported. There is extensive variance in the Laterality index (LI) calculation across the studies, and the optimal calculation method remains unclear. We attempted to calculate the LI in different ways in the same subjects, in order to find the LI calculation method with the highest correlation to the Wada test. Fifteen patients (10 females, 5 males) suffering from medically intractable temporal lobe epilepsy (TLE) (12 left, 3 right) were admitted for the study. The patients underwent a standardized bilateral intracarotid short-acting barbiturate test. Language testing included spontaneous speech, oral comprehension, reading, object and picture naming, and repetition. All the tasks were scored separately in order to increase the possibility of correlation between Wada and LI. A silent phonemic verbal fluency task (VFT) was used as a language paradigm for functional measurement. Regions of interest (ROIs), with a known association with language function (Broca’s area, the lateral prefrontal cortex, etc.), were defined. First, the LIs were calculated from the ROIs using a previously reported method (simple suprathreshold count). Next, we used several new methods of LI calculation (t–weighting of voxels, methods independent of the choice of the statistical threshold, etc.) The most significant correlation with Wada was proven in the LIs that were evaluated from Broca’s area (up to R = 0.94, P = 1 × 10⁻⁷). However, the new LI calculation methods used in the present study did not produce a statistically significant benefit in comparison to previously reported methods.     

Topography of signaling molecules as detected by electron microscopy on plasma membrane sheets isolated from non-adherent mast cells

Immunolabeling of isolated plasma membrane (PM) sheets combined with high-resolution electron microscopy is a powerful technique for understanding the topography of PM-bound signaling molecules. However, this technique has been mostly confined to analysis of membrane sheets from adherent cells. Here we present a rapid, simple and versatile method for isolation of PM sheets from non-adherent cells, and show its use for examination of the topography of Fcepsilon receptor I (FcepsilonRI) and transmembrane adaptors, LAT (linker for activation of T cells) and NTAL (non-T cell activation linker), in murine bone marrow-derived mast cells (BMMC). The data were compared with those obtained from widely used but tumor-derived rat basophilic leukemia (RBL) cells. In non-activated cells, FcepsilonRI was distributed either individually or in small clusters of comparable size in both cell types. In multivalent antigen-activated BMMC as well as RBL cells, FcepsilonRI was internalized to a similar extent, but, strikingly, internalization in BMMC was not preceded by formation of large (~200 nm) aggregates of FcepsilonRI, described previously in activated RBL cells. On the other hand, downstream adaptor proteins, LAT and NTAL, were localized in independent domains in both BMMC and RBL cells before and after FcepsilonRI triggering. The combined data demonstrate unexpected properties of FcepsilonRI signaling assemblies in BMMC and emphasize the importance of studies of PM sheets isolated from non-tumor cells.     

Morris water maze learning in Long-Evans rats is differentially affected by blockade of D1-like and D2-like dopamine receptors

Dopaminergic neurotransmission is involved in several brain functions including spatial cognition. In the present study we examine the effects of systemic administration of D1-like receptor antagonist SCH23390 and D2-like receptor antagonist sulpiride on the acquisition of the Morris water maze task. We used visible versus hidden platform versions of the MWM in order to distinguish between the effects of the drugs on the procedural versus cognitive aspects of the task. SCH23390 was found to prolong escape latencies to the visible platform at a higher dose (0.05mg/kg), whilst the lower dose (0.02mg/kg) left both procedural and cognitive functions almost unchanged. SCH23390 was also found to reduce swimming speed. Sulpiride did not affect the visible platform learning at any of three doses studied (30, 60 and 100mg/kg); the highest dose of sulpiride (100mg/kg) impaired place navigation to the hidden platform, without affecting the swim speed. The results of the present study show a difference in the involvement of D1-like and D2-like receptors in the MWM acquisition.     

Patterns of surface temperatures in two mole-rats (Bathyergidae) with different social systems as revealed by IR-thermography

Furred subterranean mammals face the problem of dissipating heat to the environment because high humidity and absence of air flow in sealed belowground tunnels constrain heat loss from body by convection and evaporation. In order to detect body areas responsible for heat loss, surface temperatures in two species of African mole-rats were measured at different ambient air temperatures by infrared thermography. Fur characteristics were also evaluated. Thinner pelage of the ventrum, its moderate temperature and large size suggest that ventral side of the body is the main thermal avenue for heat loss in both species. Interspecific differences could be explained by different fur characteristics connected with social thermoregulation. Compared to the social Fukomys mechowii, the solitary Heliophobius argenteocinereus has denser and longer fur on most of its body; its surface temperature was thus lower than in F. mechowii at lowered ambient temperatures. On the other hand, the denser and longer hair cover in H. argenteocinereus impedes heat dissipation at highest ambient temperatures (and probably also during digging activity) resulting in increase of core body temperature. H. argenteocinereus seems to be more sensitive to overheating than F. mechowii. At lower air temperatures, the social species may uses huddling to combat hypothermia.     

Atherosclerosis and vascular aging as modifiers of tumor progression, angiogenesis, and responsiveness to therapy

It is rarely considered that age-related common vascular co-morbidities may affect therapeutic outcomes of antiangiogenic therapy in cancer. Indeed, the accepted model of human disease consists of 4- to 8-week-old (young) tumor-bearing, but otherwise healthy, experimental mice, yet human cancers are diagnosed and treated in later decades of life when atherosclerosis and vascular diseases are highly prevalent. Here we present evidence that tumor growth and angiogenesis are profoundly altered in mice affected by natural aging and with genetically induced atherosclerosis (in ApoE(-/-) mice). Thus, transplantable tumors (Lewis lung carcinoma and B16F1) grew at higher rates in young (4 to 8 weeks old) ApoE(+/+) and ApoE(-/-) nonatherosclerotic syngeneic recipients than in their old (12 to 18 months old) or atherosclerotic (old/ApoE(-/-)) counterparts. These age-related changes were paralleled by reduced tumor vascularity, lower expression of tumor endothelial marker 1, increased acute tumor hypoxia, depletion of circulating CD45(-)/VEGFR(+) cells, and impaired endothelial sprouting ex vivo. Exposure of tumor-bearing mice to metronomic therapy with cyclophosphamide exerted antimitotic effects on tumors in young hosts, but this effect was reduced in atherosclerotic mice. Collectively, our results suggest that vascular aging and disease may affect tumor progression, angiogenesis, and responses to therapy.