Der histochemische Eisennachweis

Zusammenfassung Der histochemische Eisennachweis nachTirmann undSchmelzer erfaßt nicht das gesamte reaktionsfähige Gewebseisen. Daher ist Eisen in normalen Organen nicht nachweisbar.An Stelle von Kaliumferricyanid muß bei dem Eisennachweis Kaliumferrocyanid verwendet werden, um rückgebildetes Eisen-III-oxydhydrat in Eisenblau umzuwandeln.Sicher und vollständig wird das reaktionsfähige Eisen in H₂S-Gefrierschnitten erfaßt. Es ist auch in normalen Organen: Leber, Niere u. a. nachweisbar.Mit 3 TextabbildungenMit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Scrota1 melanins in bats (Chiroptera): description, distribution and function

Several species of bats contain pigment granules within the scrotal skin, tunica vaginalis, or tunica albuginea surrounding the testis and/or epididymis. Seventy-two species of bats, representing 49 genera were examined for the presence of such pigmentation. Histological, chemical and spectrophotometric tests were performed and confirmed the pigment as melanin. Melanin was found only in the families Pteropidae, Megadermatidae, Myzopodidae and Vespertilionidae. A strong correlation exists between scrotal pigmentation and roosting in locations where the bats are exposed to solar radiation. Melanin pigmentation in the scrotal region appears to be an adaption protecting male germinal tissue from the harmful effects of ultraviolet radiation. In one species, Laoiu frons , melanin deposited within the scrotal skin appears to have a social/reproductive communication function.     

Identification of CLEC12B, an inhibitory receptor on myeloid cells

Activation of immune cells has to be tightly controlled to prevent detrimental hyperactivation. In this regulatory process molecules of the C-type lectin-like family play a central role. Here we describe a new member of this family, CLEC12B. The extracellular domain of CLEC12B shows considerable homology to the activating natural killer cell receptor NKG2D, but unlike NKG2D, CLEC12B contains an immunoreceptor tyrosine-based inhibition motif in its intracellular domain. Despite the homology, CLEC12B does not appear to bind NKG2D ligands and therefore does not represent the inhibitory counterpart of NKG2D. However, CLEC12B has the ability to counteract NKG2D-mediated signaling, and we show that this function is dependent on the immunoreceptor tyrosine-based inhibition motif and the recruitment of the phosphatases SHP-1 and SHP-2. Using monoclonal anti-CLEC12B antibodies we found de novo expression of this receptor on in vitro generated human macrophages and on the human myelo-monocytic cell line U937 upon phorbol 12-myristate 13-acetate treatment, suggesting that this receptor plays a role in myeloid cell function.     

Indicative profundal oligochaete assemblages in selected small Swedish lakes

Fifty-seven small- to middle-size lakes of Sweden (size-range from <0.1 to about 200 km²) representing different regions and lake-types were selected for the purpose of demonstrating the indicative value of profundal oligochaete communities also of relatively small lakes. It is well known since previously that profundal oligochaete assemblages of large lakes in Europe may be particularly indicative of the current trophic situation, but little has been known about smaller lakes in this respect. Characterizations based on oligochaete community structure in these lakes could be summarized in eight different groups: Characteristic oligochaete communities in oligotrophic lakes of relatively small size are Spirosperma ferox, Stylodrilus heringianus and Tubifex tubifex. Oligochaete species richness is low (1–3 species) and so is species abundance (<300 specimens m^–2). Very eutrophic lakes of the same size-range are largely dominated by the tubificids Limnodrilus hoffmeisteri and/or Potamothrix hammoniensis. Substitute or complementary species may be T. tubifex. Species richness is usually low (1–3 species), but abundance is high to very high (>5000 specimens m^–2). Mesotrophic to moderately eutrophic lakes of the same size categories contain more species than the above extremes, most of them being fairly tolerant to oxygene deficiency such as Tubifex ignotus, a number of species of the genera Aulodrilus and Limnodrilus and a variety of so-called Ponto-Caspian Potamothrix species, most of them having recently invaded south-eastern Sweden. Species richness is moderate to high (4–8 species), while abundance may vary considerably (range from about 300 to about 5000 specimens m^–2). In this respect, small- to middle-size lakes do not differ much from larger lakes. For the trophic characterization of lake water the mean concentrations of total-phosphorus (in g l^–1) is generally regarded to be the most appropriate measure. The match between characterizations based upon oligochaete community structure and total-phosphorus concentrations is generally very good for large lakes. The present study shows that this applies fairly well also to the majority of the small- to middle-size lakes selected for this study. This has not been shown before. Obvious deviations from this rule could be caused by a scarcity of reliable chemical data, or considerable seasonal shifts in the chemical composition ot the water, which in its turn may be caused by short water residence time. Littoral elements in the fauna, which are generally less indicative, or effects of local inflows may also be more frequent here than in lakes of larger size.     

A new species of small-eared shrew in the <Emphasis Type="Italic">Cryptotis thomasi</Emphasis> species group from Costa Rica (Mammalia: Eulipotyphla: Soricidae)

We describe a new species of small-eared shrew, genus Cryptotis Pomel, 1848 (Eulipotyphla: Soricidae), from near the community of Monteverde in the Tilarán highlands of northwestern Costa Rica. The new species is immediately distinguished from all other Costa Rican shrews its large size and long tail. Morphologically, it belongs to the Cryptotis thomasi group of small-eared shrews, a clade that is more typically distributed in the Andes Cordillera and other highland regions of northern South America. The new Costa Rican species and the Panamanian endemic Cryptotis endersi Setzer, 1950 are the only two members of this species group known to occur in Central America. Like most other members of the C. thomasi group for which the postcranial skeleton has been studied, the new species tends be more ambulatory (rather than semi-fossorial) when compared with other members of the genus. Our survey efforts over several decades failed to locate a population of the new species, and we discuss its conservation status in light of its limited potential distribution in the Tilarán highlands and the significant climatic change that has been documented in the Monteverde region during the past four decades.     

All-In-One: Advanced preparation of Human Parenchymal and Non-Parenchymal Liver Cells

Background & Aims

Liver cells are key players in innate immunity. Thus, studying primary isolated liver cells is necessary for determining their role in liver physiology and pathophysiology. In particular, the quantity and quality of isolated cells are crucial to their function. Our aim was to isolate a large quantity of high-quality human parenchymal and non-parenchymal cells from a single liver specimen.

Methods

Hepatocytes, Kupffer cells, liver sinusoidal endothelial cells, and stellate cells were isolated from liver tissues by collagenase perfusion in combination with low-speed centrifugation, density gradient centrifugation, and magnetic-activated cell sorting. The purity and functionality of cultured cell populations were controlled by determining their morphology, discriminative cell marker expression, and functional activity.

Results

Cell preparation yielded the following cell counts per gram of liver tissue: 2.0±0.4×10⁷ hepatocytes, 1.8±0.5×10⁶ Kupffer cells, 4.3±1.9×10⁵ liver sinusoidal endothelial cells, and 3.2±0.5×10⁵ stellate cells. Hepatocytes were identified by albumin (95.5±1.7%) and exhibited time-dependent activity of cytochrome P450 enzymes. Kupffer cells expressed CD68 (94.5±1.2%) and exhibited phagocytic activity, as determined with 1μm latex beads. Endothelial cells were CD146⁺ (97.8±1.1%) and exhibited efficient uptake of acetylated low-density lipoprotein. Hepatic stellate cells were identified by the expression of α-smooth muscle actin (97.1±1.5%). These cells further exhibited retinol (vitamin A)-mediated autofluorescence.

Conclusions

Our isolation procedure for primary parenchymal and non-parenchymal liver cells resulted in cell populations of high purity and quality, with retained physiological functionality in vitro. Thus, this system may provide a valuable tool for determining liver function and disease.     

Protein kinase MARK/PAR-1 is required for neurite outgrowth and establishment of neuronal polarity

Protein kinases of the microtubule affinity-regulating kinase (MARK) family were originally discovered because of their ability to phosphorylate certain sites in tau protein (KXGS motifs in the repeat domain). This type of phosphorylation is enhanced in abnormal tau from Alzheimer brain tissue and causes the detachment of tau from microtubules. MARK-related kinases (PAR-1 and KIN1) occur in various organisms and are involved in establishing and maintaining cell polarity. Herein, we report the ability of MARK2 to affect the differentiation and outgrowth of cell processes from neuroblastoma and other cell models. MARK2 phosphorylates tau protein at the KXGS motifs; this results in the detachment of tau from microtubules and their destabilization. The formation of neurites in N2a cells is blocked if MARK2 is inactivated, either by transfecting a dominant negative mutant, or by MARK2 inhibitors such as hymenialdisine. Alternatively, neurites are blocked if the target KXGS motifs on tau are rendered nonphosphorylatable by point mutations. The results suggest that MARK2 contributes to the plasticity of microtubules needed for neuronal polarity and the growth of neurites.     

Can vegetation be discretely classified in species‐poor environments? Testing plant community concepts for vegetation monitoring on sub‐Antarctic Marion Island

The updating and rethinking of vegetation classifications is important for ecosystem monitoring in a rapidly changing world, where the distribution of vegetation is changing. The general assumption that discrete and persistent plant communities exist that can be monitored efficiently, is rarely tested before undertaking a classification. Marion Island (MI) is comprised of species‐poor vegetation undergoing rapid environmental change. It presents a unique opportunity to test the ability to discretely classify species‐poor vegetation with recently developed objective classification techniques and relate it to previous classifications. We classified vascular species data of 476 plots sampled across MI, using Ward hierarchical clustering, divisive analysis clustering, non‐hierarchical kmeans and partitioning around medoids. Internal cluster validation was performed using silhouette widths, Dunn index, connectivity of clusters and gap statistic. Indicator species analyses were also conducted on the best performing clustering methods. We evaluated the outputs against previously classified units. Ward clustering performed the best, with the highest average silhouette width and Dunn index, as well as the lowest connectivity. The number of clusters differed amongst the clustering methods, but most validation measures, including for Ward clustering, indicated that two and three clusters are the best fit for the data. However, all classification methods produced weakly separated, highly connected clusters with low compactness and low fidelity and specificity to clusters. There was no particularly robust and effective classification outcome that could group plots into previously suggested vegetation units based on species composition alone. The relatively recent age (c. 450,000 years B.P.), glaciation history (last glacial maximum 34,500 years B.P.) and isolation of the sub‐Antarctic islands may have hindered the development of strong vascular plant species assemblages with discrete boundaries. Discrete classification at the community‐level using species composition may not be suitable in such species‐poor environments. Species‐level, rather than community‐level, monitoring may thus be more appropriate in species‐poor environments, aligning with continuum theory rather than community theory.