Die Reizwirkung des Methylcholanthrens auf die Melanocyten der Haut bei verschiedenen Mäusestämmen

Zusammenfassung Die Haut gefärbter Mäuse ist im Gegensatz zu den Haaren normalerweise unpigmentiert. Durch Behandlung mit Methylcholanthren können aber die in der Haut enthaltenen Pigmentzellen (Melanocyten) aktiviert und zur Melaninproduktion angeregt werden. Diese Aktivierung verläuft bei allen Mäusestämmen gleich. Sie beginnt bei den Melanocyten der Haarwurzeln und der Epidermis und greift dann bei weiterer Pinselung mit Methylcholanthren auch auf diejenigen der Cutis über. Die Farbe des gebildeten Pigments hängt dabei von den vorhandenen Farbfaktoren ab.Unterbricht man die Behandlung, so stellen die Melanocyten die Pigmentproduktion wieder ein, können aber durch erneute Methylcholanthren-Pinselung jederzeit reaktiviert werden.Als Folge des Entzündungsreizes beobachtet man ferner eine lokale Anreicherung von Freßzellen, die ebenfalls reversibel ist. Diese Histiocyten phagocytieren das überzählig gebildete Pigment.Zwischen der Reizwirkung und der cancerogenen Wirkung des Methylcholanthrens besteht keine Korrelation. Erstere verläuft, wie gesagt, bei allen Mäusen gleich, während die Krebserzeugung durch Methylcholanthren hauptsächlich von der genetischen Konstitution der verwendeten Tiere abhängig ist (Brenner 1958).     

Effect of sex and gonadal hormones on rat plasma lipids during the development of an essential fatty acid deficiency

1. Male, female and castrated rats treated with oestradiol (30mug./week) or testosterone (2mg./week) were given an essential fatty acid-deficient diet containing 10% of hydrogenated coconut oil for 9 weeks. The concentrations and fatty acid composition of plasma phospholipids, cholesteryl esters and triglycerides were determined. 2. Between the second and third weeks of the deficiency, concentrations of plasma cholesteryl esters, phospholipids and triglycerides decreased, then remained relatively constant. There were no significant differences between males and females, but oestradiol caused a significant rise in plasma phospholipids and triglycerides as compared with testosterone-treated animals. 3. During the first 2 weeks of the deficiency, linoleic acid in the plasma lipids of all groups decreased to low concentrations and changed very little thereafter. 4. Female rats maintained higher percentages and concentrations of arachidonic acid and stearic acid in plasma phospholipids and arachidonic acid in cholesteryl esters than did males. Males had higher proportions of eicosatrienoic acid and oleic acid. There was no sex difference in the fatty acid composition of plasma triglycerides. 5. Oestradiol-treated rats had concentrations of cholesteryl and phospholipid arachidonate comparable with those of female rats and higher than the testosterone-treated group. Eicosatrienoic acid in the oestradiol-treated rats was high and resembled that of the male rats, apparently because of the higher concentration of plasma phospho lipids in this group. 6. Supplementation of the essential fatty acid-deficient rats with linoleate restored plasma cholesteryl and phospholipid linoleate and arachidonate nearly to normal concentrations in a single day. The increase in arachidonic acid in these fractions was accompanied by a similar quantitative decrease in eicosatrienoic acid. 7. These sex differences appear to be related to the smaller size of the female rat and to a more direct influence of oestradiol on the formation or maintenance of phospholipids rich in arachidonic acid.     

Increased stiffness of the rat liver precedes matrix deposition: implications for fibrosis

Liver fibrosis, the response to chronic liver injury, results from the activation of mesenchymal cells to fibrogenic myofibroblasts. We have recently shown that two key myofibroblast precursor populations, hepatic stellate cells and portal fibroblasts, undergo activation in culture in response to increasing substrate stiffness. We therefore hypothesized that alterations in liver stiffness precede myofibroblast activation and fibrosis in vivo as well. To test this hypothesis, we induced fibrosis in rats by twice weekly injections of carbon tetrachloride (CCl(4)) and then killed the animals at various time points ranging from 3 to 70 days after the initiation of injury. The shear storage modulus of the whole liver was measured on fresh tissue; fixed and frozen tissue from the same livers was used to quantify fibrosis. We observed that liver stiffness increased immediately and continued to increase, leveling out by day 28. Fibrosis, measured histologically by trichrome staining as well as by quantitative sirius red staining, increased with time, although these increases were delayed relative to changes in stiffness. There was no direct correlation between stiffness and fibrosis at early or late time points. Treatment of a second cohort of rats with the lysyl oxidase inhibitor, beta-aminopropionitrile (BAPN), partially prevented early increases in liver stiffness. We concluded that increases in liver stiffness precede fibrosis and potentially myofibroblast activation. Liver stiffness appears to result from matrix cross-linking and possibly other unknown variables in addition to matrix quantity. We suggest that increased liver stiffness may play an important role in initiating the early stages of fibrosis.     

Management of modern agricultural landscapes increases nest predation rates in Black‐tailed Godwits Limosa limosa

Effective conservation of endangered species requires a solid understanding of the demographic causes of population change. Bird populations breeding on agricultural grasslands have declined because their preferred habitat of herb‐rich meadows has been replaced by grassland monocultures. The timing of agricultural activities in these monocultural grasslands is critical, as they often coincide with the nesting phase of breeding birds. Here, we aim to identify the effect of habitat management and targeted nest protection on nest survival of Black‐tailed Godwits Limosa limosa in the Netherlands, a population that has shown a 70% reduction in breeding population size since the 1970s. To protect nests in monocultures from destruction, farmers are paid to either delay mowing or leave a patch of unmown grass around the nest, a patch which in practice varied in size. In herb‐rich meadows, which are typically managed for bird conservation purposes, mowing occurs after hatching. Nest survival declined as the season advanced, more steeply on monocultures than on meadows. Targeted nest protection was only partially successful, as nest predation was considerably higher on mown grassland monocultures with small unmown patches around the nest than in mown monocultures with large unmown patches and in unmown fields. Increased predator densities over the years have been suggested as an important cause of the trend towards lower nest survival, but here we show that nest survival was higher on herb‐rich meadows than on monocultures, and similar to the 1980s. It thus seems that increased predator densities are an increased threat during the egg stage only if habitat quality is low. High‐quality habitat in the form of herb‐rich meadows therefore provides a degree of protection against predators.     

A second European collaborative study on polymerase chain reaction for Toxoplasma gondii, involving 15 teams

In order to investigate the accuracy and practicability of the polymerase chain reaction (PCR) in the antenatal diagnosis of congenital toxoplasmosis, a collaborative study involving 15 European laboratories was performed under the auspices of the Biomed 2 Programme of the European Community. Each team received 12 aliquots (four negative, eight positive) of `artificial samples' made of amniotic fluid spiked with tachyzoites of the RH strain of Toxoplasma gondii. Each team performed its own PCR protocol (all were different). Nine of the 15 laboratories were able to detect a single parasite, but two of the 15 found all samples negative. Four of the 15 laboratories found one or more control samples to be falsely positive. This study highlights the lack of homogeneity between PCR protocols and performance and underlines the need for an external quality assurance scheme which could provide `reference' samples that could be used by any laboratory wanting to establish and maintain an accurate diagnostic test based on PCR.     

The annexins of Dictyostelium

Annexins are a highly conserved ubiquitous family of Ca2+- and phospholipid-binding proteins present in nearly all eukaryotic cells. Analysis of the Dictyostelium genome revealed the presence of two annexin genes, the annexin C1 gene (nxnA) giving rise to two isoforms of 47 and 51 kDa (previously synexin), and the annexin C2 gene (nxnB) coding for a 56-kDa protein with 33% sequence identity to annexin C1. Annexin C2 is expressed at very low and constant levels throughout development. Quantification by real-time PCR indicated that it is present in about 35-fold lower amounts compared to annexin C1. We have used a GFP-tagged annexin C2 to study its cellular distribution and dynamics. In cell fractionation studies, annexin C2 cofractionates with annexin C1 and is enriched in the 100,000 g pellet. Like annexin C1, GFP-AnxC2 stains the plasma membrane. In addition it is present in the perinuclear region and overlaps to some degree with the Golgi apparatus, whereas annexin C1 is present on intracellular membranes resembling endosomal membranes and in the nucleus. Annexin C2 is not observed in the nucleus. An annexin C1 mutant (SYN-) which shows a defect during multicellular development can be rescued by full-length annexin C1, whereas overexpression of GFP-AnxC2 did not rescue the developmental defect The data support the concept that annexins, although having a highly conserved structure, participate in different functions in a cell.     

Functional anatomy of five endangered tropical timber wood species of the family Dipterocarpaceae

Wood anatomy of five dipterocarp species endemic to the Philippines was studied with the goal to explore functional wood traits of ecological significance. Stem wood of 6-year-old trees grown under similar environmental conditions in a plantation (Leyte, Philippines) was used. Wood densities decreased in the following order Hopea plagata > Dipterocarpus kerrii > Parashorea malaanoman > Shorea almon ≈ Shorea contorta. This was mainly caused by significantly thicker fiber cell walls of H. plagata and D. kerrii than those of the other three species. Wood density was negatively correlated with the abundance of axial parenchyma cells. Predicted conductance was independent from wood density and lowest in H. plataga and highest in D. kerrii and S. contorta. These results indicate that H. plagata and D. kerrii is woods have higher constructions costs in term of carbon per unit of biomass, and that H. plagata is probably better acclimated to varying soil moisture than the other species.     

Mucosa‐associated biohydrogenating microbes protect the simulated colon microbiome from stress associated with high concentrations of poly‐unsaturated fat

Polyunsaturated fatty acids (PUFAs) may affect colon microbiome homeostasis by exerting (specific) antimicrobial effects and/or interfering with mucosal biofilm formation at the gut mucosal interface. We used standardized batch incubations and the Mucosal‐Simulator of the Human Microbial Intestinal Ecosystem (M‐SHIME) to show the in vitro luminal and mucosal effects of the main PUFA in the Western diet, linoleic acid (LA). High concentrations of LA were found to decrease butyrate production and Faecalibacterium prausnitzii numbers dependent on LA biohydrogenation to vaccenic acid (VA) and stearic acid (SA). In faecal batch incubations, LA biohydrogenation and butyrate production were positively correlated and SA did not inhibit butyrate production. In the M‐SHIME, addition of a mucosal environment stimulated biohydrogenation to SA and protected F. prausnitzii from inhibition by LA. This was probably due to the preference of two biohydrogenating genera Roseburia and Pseudobutyrivibrio for the mucosal niche. Co‐culture batch incubations using Roseburia hominis and F. prausnitzii validated these observations. Correlations networks further uncovered the central role of Roseburia and Pseudobutyrivibrio in protecting luminal and mucosal SHIME microbiota from LA‐induced stress. Our results confirm how cross‐shielding interactions provide resilience to the microbiome and demonstrate the importance of biohydrogenating, mucosal bacteria for recovery from LA stress.