Edible mushroom-related poisoning: A study on circumstances of mushroom collection, transport, and storage.

BACKGROUND: The American Association of Poison Control Center (AAPCC) shows that in 2012 there were 0.3% of human exposures involving mushrooms. Only 17% of 6600 cases were then identified by the species. The present retrospective study was designed to identify the epidemiology of mushroom poisoning in adults admitted to Krakow's Department of Clinical Toxicology (DCT) from 2002 to 2009. MATERIALS AND METHODS: This study was conducted retrospectively after examining the files of 457 adult patients with wild mushroom poisoning. Mycological analysis was made and the species of the poisoning-inducing mushroom was determined. Furthermore, the circumstances related to the mushroom gathering, transport, storage, preparation, and consumption have been analyzed. RESULTS: The analysis revealed that in 400 (87.53%) out of 457 cases, the clinical symptoms were caused by ingestion of identified edible mushroom species. The main reason for edible mushroom poisoning is associated with their incorrect processing after harvest. The analysis of the circumstances of mushroom collection, transport, and storage shows that the largest percentage of poisoning was connected with long-term storage of mushroom dishes, collecting, and storing them in plastic bags, and long storage of mushrooms. CONCLUSION: Based on spore analysis of the gastric content, edible mushrooms were responsible for the great majority of mushroom poisoning cases admitted to the DCT. The toxicity of edible mushroom is associated with proceeding with them during collection, transport, and storage. The medical history should be supplemented by questions concerning these circumstances. The identification of the mushroom by a mycologist is highly desirable.

Lipid content and cryotolerance of porcine embryos cultured with phenazine ethosulfate.

The addition of phenazine ethosulfate (PES) to culture medium was investigated for its effect on pig embryo development, apoptosis, cytoplasmic lipid content and survival after OPS vitrification. Porcine zygotes were cultured in NCSU-23 medium supplemented with 0 (control) or 0.05 microM PES up to the blastocyst stage and were vitrified using OPS technology. Culture of embryos with PES reduced the cytoplasmic lipid content, as measured by fluorescence of blastocysts stained with Nile Red. The survival rate of vitrified blastocysts was slightly enhanced, although not significantly, in the presence of PES compared to the PES-free group (45.2 and 37.9 percent, respectively). These results showed that culturing porcine embryos in medium with PES increased the proportion of morula and blastocyst formation and reduced the index of DNA fragmentation and the cytoplasmic lipid content of cultured blastocysts. However, the use of PES during in vitro culture had limited effect on porcine blastocyst survival after vitrification.

Mitochondrial activity and morphology in developing porcine oocytes and pre-implantation non-cultured and cultured embryos.

Mitochondria are important determinants of developmental competence for oocytes and embryos owing to their central role in cellular metabolism, yet mitochondrial activity and morphometry during early porcine development have not been quantified. In this study, we examined the membrane potential Δψ(m) and the surface density Sv(in,m) of the inner mitochondrial membrane in pig oocytes and pre-implantation embryos using fluorescent probes and confocal microscopy. Mitochondria and their cristae were also examined by transmission electron microscope. Δψ(m) was consistently low from immature oocytes up to morulae and increased significantly in the early blastocyst before decreasing at the expanded blastocyst stage. This stage-dependent pattern of Δψ(m) changes differs from that reported for other mammals. We also determined that Δψ(m) is lower in cultured when compared to non-cultured porcine early blastocysts. Sv(in,m) was higher in immature oocytes than mature oocytes and remained constant up to the 4- to 8-cell embryo stage. It increased significantly at morula and early blastocyst stages. No differences in Sv(in,m) were found between developmentally matched non-cultured and cultured embryos. These results indicate that the inner mitochondrial membrane potential and surface density change significantly during pre-implantation porcine development in relation to metabolic alterations of the embryo. It is possible that modification of Δψ(m) by manipulating culture conditions may improve the performance of embryos that develop in vitro.

Quantification of connexin43 gap junctions in porcine myometrium by confocal microscopy and stereology.

The aim of this study was to quantify the number and size of connexin43 (Cx43) gap junctions in the circular and longitudinal layers of myometrium of the non-pregnant pig. We developed a novel approach to measure the mean surface area (s), numerical density (N(v)) and surface density (S(v)) of gap junctions using confocal microscopy and stereological analysis. Immunolabelled Cx43 gap junctions were measured in the subendometrial and deep regions of the circular layer and in the longitudinal layer of the myometrium of pre-pubertal pig and mature pig at pre-ovulatory and secretory stages of the oestrous cycle. In the circular subendometrial region, all investigated stereological parameters of Cx43 gap junctions (s, N(v) and S(v)) were significantly higher (p<0.05) than those of the circular deep region and the longitudinal layer in all three groups of animals. These results indicate the large-scale heterogeneity of the number and size of Cx43 gap junctions across the myometrium in non-pregnant pig and emphasize the existence of functional diversity among myometrial cells.

Changes of lipid composition in non-cultured and cultured porcine embryos.

The objective of the present study was to evaluate the lipid composition of cultured and non-cultured pig embryos during cleavage using histochemical methods. The authors studied pig zygotes as well as 2-to 4-cell embryos, morulae, and blastocysts that were either non-cultured or cultured in NCSU-23 medium. To detect different types of lipids, the authors used the Churukian method with Oil red O, the Sudan black B method, the Cain method with Nile blue sulfate, and the modified osmium tetroxide-ethanol treatment. In the zygotic lipid droplets, diverse classes of unsaturated and saturated lipids were found, with particularly high levels of unsaturated hydrophobic lipids, mainly triglycerides and other esters, free fatty acids, and phospholipids. In the zygotic cytoplasm, the authors observed high levels of fatty acids and phospholipids. The total lipid content remained constant up to the morula stage, decreasing later at the blastocyst stage, but the overall amount of unsaturated lipids declined earlier, at the 2-to 4-cell stage. The amount of free fatty acids and phospholipids decreased during cleavage in both non-cultured and cultured porcine embryos. The main differences between the non-cultured and cultured embryos were the more pronounced reduction in the amount of unsaturated hydrophobic lipids in droplets and the cytoplasmic free fatty acids observed in the cultured morula and the lower content of phospholipids in the cytoplasm of the cultured 2-to 4-cell embryos relative to the non-cultured embryos. The decrease in the unsaturated lipid, free fatty acid, and phospholipid content during in vivo development and the differences in the amount of these types of lipids between developmentally matched cultured and non-cultured pig embryos correlate well with modifications of lipid and carbohydrate metabolism.

Lipid content of non-cultured and cultured pig embryo.

The objectives of the study were: (i) to work out a precise and efficient method for quantitative analysis of lipid content and (ii) to quantitatively determine the lipid content in non-cultured and cultured pig embryos. The experiment was carried out on pig embryos from zygote to late blastocyst stages produced in vivo and embryos collected at the zygote stage and then cultured in vitro up to blastocyst stage. Embryos were fixed, dehydrated, embedded in epoxy resin and cut into semi-thin sections to analyse the quantity of lipids in fat droplets. Stained sections were then analysed with Cavalieri and point counting methods to evaluate the following stereological parameters of the embryo: total embryo volume - V(e), volume density of cytoplasm per unit volume of embryo - Vv(c,e), volume density of lipid droplets per unit volume of embryo cytoplasm - Vv(fat,c) and total volume of lipid droplets per whole embryo - V(fat). Values of Vv(fat,c) and V(fat) remained unchanged up to the morula stage, but decreased significantly at blastocyst and late blastocyst stages both in cultured and non-cultured embryos. Volume density of lipid droplets per unit volume of embryo cytoplasm and total volume of lipid droplets for cultured embryos showed statistically significant differences between late blastocyst and almost all other stages. Comparisons of Vv(fat,c) in embryos at the same stages of development but differing in origin of embryos (non-cultured or cultured) show that statistically significant differences exist for all analysed stages. In conclusion, differences in lipid content observed in pig embryos were dependent on the developmental stage of the embryo as well as the culture conditions (i.e. cultured and non-cultured embryos at the same stage of development).

Do germ line cells in Allacma fusca (Insecta, Collembola, Symphypleona) have a higher metabolic rate than somatic cells.

Stereological analysis of the ultrastructure of primordial germ cells (PGCs) and the somatic (ectoderm) cells in two developmental stages of embryos and freshly hatched juveniles of Allacma fusca have shown great differences in mitochondria volume density (vd) between the two types of cells. In younger embryos (migration phase of the PGCs) the vd of mitochondria in the cytoplasm of the PGCs is 74.64% higher than in the ectoderm cells. In older embryos, (PGCs in the gonads) the vd of mitochondria is 123% higher than the corresponding value for the somatic cells cytoplasm. In the juvenile the vd of mitochondria in the ectoderm cells grows twice but is still only 2/3 of the value for the PGCs. On the basis of papers describing a direct relationship between stereological and physiological results the authors conclude that the metabolism of the primordial germ cells during embryonic development of Allacma fusca is much higher than that of the somatic ones. If the above conclusion is correct, the results presented here confirm the "disposable soma theory" (Kirkwood & Holliday 1979).

Utilization of yolk platelets during early embryonic development of Rana temporaria and Bufo bufo.

Utilization of yolk platelets in cleaving embryos of Rana temporaria and Bufo bufo was studied by different methods. Morphological observations of yolk platelets of R. temporaria embryos at tail bud stage by transmission electron microscopy indicated four initial phases of platelet degradation. The pattern of these events is similar to that found in embryos of B. bufo. The morphological observations were confirmed by energy-dispersive X-ray microanalysis of the elemental content of platelets and by selected-area electron diffraction of platelet cores. Covalently bound sulphur content decreased during cleavage and the content of different inorganic ions changed, whereas the structure of crystalline core remained constant. Morphological changes found in the amorphous cortex of yolk platelets were due to their utilization. Stereological measurements indicated that utilization during cleavage increased, but only the initial phases of yolk platelet degradation were seen. The volume of the cortex did not decrease and the crystalline core did not fragment.

Spatial distribution of yolk platelets and fat droplets in oocytes and cleaving embryos of the common frog (Rana temporaria) and toad (Bufo bufo).

Animal-vegetal gradients of fat droplets and yolk platelets have been quantitatively determined in mature oocytes and in cleaving embryos in two anuran species Rana temporaria and Bufo bufo using stereological methods. Volume densities Vv which describe properly the amount of nutrients in embryos have been used during the measurements. The embryos till the stage of late blastula are spherical: the spherical form is not distorted even by blastocoel forming eccentrically, in the animal hemisphere. Increased diameters during cleavage, which have been observed in the two species, can be ascribed to growth of the blastocoel and increased number of blastomeres while the quantity of the cytoplasm does not increase. The volume densities of yolk platelets and fat droplets per unit of cytoplasm volume remains constant throughout the cleavage. Volume densities change along the animal-vegetal axis in accordance with the course of the 3rd polynomial depending on the distance of the area under study from the vegetative pole f: y-->Vv, f = ay3 + by2 + cy + d. Parameters of fitting functions a, b, c and d change considerably during development of the embryo, which proves reorganisation of the cytoplasm during cleavage: the most significant changes occur from fertilisation to the four-cell stage and during growth of the blastocoel from morula to late blastula stage. Distortion of the axial symmetry of yolk platelet distribution in embryos occurring after fertilisation is not extensive in the two studied species. The mathematical model of spatial distribution of yolk platelets and fat droplets in oocytes and cleaving embryos in the studied anuran species was proposed using the above data.

The lipoprotein crystals in yolk platelets of a shark, Squalus ocanthias (Selachii).

Lipovitellin-phosvitin crystals in yolk platelets from the egg of a shark, Squalus ocanthias (Selachii), were studied by the selected area electron diffraction method. Ultra-thin sections of yolk platelets possess an orthorhombic lattice with lattice parameters: a = 8.0 nm, b = 15.9 nm, and c = 18.1 nm. Comparative and quality analysis of the reflection intensities of electron diffraction patterns between shark and other vertebrate yolk platelets suggest evolutionary variability of the lipovitellin structure. The lattice and its parameters, however, are highly conserved. The results of the present study fill the last gap in the comparative yolk platelet research which has been carried out over many years for exponents of all vertebrate taxonomic groups, from cyclostomes to reptiles.