An examination of the potential role of spider digestive proteases as a causative factor in spider bite necrosis

Tissue necrosis following spider bites is a widespread problem. In the continental United States, the brown recluse (Loxosceles reclusa), hobo spider (Tegenaria agrestis), garden spider (Argiope aurantia) and Chiracanthium species, among others, reportedly cause such lesions. The exact mechanism producing such lesions is controversial. There is evidence for both venom sphingomyelinase and spider digestive collagenases. We have examined the role of spider digestive proteases in spider bite necrosis. The digestive fluid of A. aurantia was assayed for its ability to cleave a variety of connective tissue proteins, including collagen. Having confirmed that the fluid has collagenases, the digestive fluid was injected into the skin of rabbits to observe whether it would cause necrotic lesions. It did not. The data do not support the suggestions that spider digestive collagenases have a primary role in spider bite necrosis.     

Time‐Continuous Phosphorus Flows in the Indian Agri‐Food Sector: Long‐Term Drivers and Management Options

Phosphorus (P) is a major agricultural nutrient and, in its mineable form, a potentially scarce resource. Countries with limited physical access to P should hence develop an effective national P governance. This requires analyses of trends and variations in P flows and stocks over time. Here, we present a long‐term P flow analysis for the Indian agri‐food sector from 1988 to 2011. Major P flows are imports of mineral P, fertilizer application, and uptake of animal fodder. The mineral P import dependency ratio is constant at around 93%. On average, 20% of P inputs to soils are lost through erosion. Key drivers of changes in P flows include population growth, dietary change, and agricultural intensification. To reduce its P fertilizer import dependence, India could, for example, substitute up to 19% of the presently applied mineral P if manure used as a household fuel were recycled, and up to 21% if P was fully recovered from wastewater and household waste. Comparing selected indicators for P use in agriculture with China and the European Union (EU) reveals that there are structural similarities, such as increasing fertilizer application rates and P accumulation in soils, with the first but large differences compared to the latter. The analyses highlight that in contrast to static indicators, the time‐continuous tracking of P flows provides substantial advantages, such as the identification of long‐term trends, drivers, and intervention options for sustainable P management, given that it allows for the interpretation of present indicators in the context of past trends and legacies.     

The EndoPredict Gene-Expression Assay in Clinical Practice - Performance and Impact on Clinical Decisions

The validated EndoPredict assay is a novel tool to predict the risk of metastases of patients with estrogen receptor positive, HER2 negative breast cancer treated with endocrine therapy alone. It has been designed to integrate genomic and clinical information and includes clinico-pathological factors such as tumor size and nodal status. The test is feasible in a decentral setting in molecular pathology laboratories. In this project, we investigated the performance of this test in clinical practice, and performed a retrospective evaluation of its impact on treatment decisions in breast cancer. During one year, EndoPredict assays from 167 patients could be successfully performed. For retrospective evaluation of treatment decisions, a questionnaire was sent to the clinical partner. Regarding the molecular EP class, samples from 56 patients (33.5%) had a low-risk, whereas 111 patients (66.5%) showed a high-risk gene profile. After integration of the clinicopathological factors the combined clinical and molecular score (EPclin) resulted in a low-risk group of 77 patients (46.4%), while 89 (53.6%) had a high risk EPclin score. The EPclin-based estimated median 10-year-risk for metastases with endocrine therapy alone was 11% for the whole cohort. The median handling time averaged three days (range: 0 to 11 days), 59.3% of the tests could be performed in three or less than three days. Comparison of pre- and post-test therapy decisions showed a change of therapy in 37.7% of patients. 16 patients (12.3%) had a change to an additional chemotherapy while 25.4% of patients (n = 33) changed to an endocrine therapy alone. In 73 patients (56.2%) no change of therapy resulted. In 6.1% of patients (n = 8), the patients did not agree to the recommendation of the tumor board. Our results show that the EndoPredict assay could be routinely performed in decentral molecular pathology laboratories and the results markedly change treatment decisions.     

Diploid and polyploid cytotype distribution in Melampodium cinereum and M. leucanthum (Asteraceae, Heliantheae)

Previous chromosomal studies within Melampodium (Asteraceae, Heliantheae) of Mexico and Central America have documented chromosome numbers n = 9, 10, 11, 12, 18, 20, 23, 25 ± 1, 27, 30, and 33. Some species also have been shown to exhibit infra- and interpopulational polyploidy. The presence of cytotype mixtures is especially pronounced in the white-rayed complex, which occurs in the southwestern United States and adjacent Mexico. This group includes M. cinereum (n = 10 and 20), M. leucanthum (n = 10 and 20), and M. argophyllum (n = 30). Cytotype distribution has been newly analyzed in 415 plants from 152 populations and added to existing data from 185 plants from 113 populations, yielding information from a total of 600 individuals from 265 populations. Within M. cinereum and M. leucanthum are parapatric distributions of cytotypes, with tetraploids centered in the eastern and diploids in the western portions of their ranges. Tetraploids are most likely of autopolyploid origin, forming recurrently, with adaptations that allow colonization and establishment in new ecological regions. Contact zones are relatively narrow and only two triploid individuals have been detected. The tetraploid cytotypes probably extended eastward into central and southern Texas to the natural barriers at the edge of the Edward's Plateau in M. leucanthum and the low sandy plains in M. cinereum. The hexaploid M. argophyllum is interpreted as a relict surviving in the low mountains of northern Mexico; it may be an allopolyploid of hybrid origin between ancestors of the evolutionary lines that eventually yielded M. cinereum and M. leucanthum.     

Morphogenesis of the antenna of the male silkmoth, Antheraea polyphemus. IV. Segmentation and branch formation

The imaginal antenna of the male silkmoth Antheraea polyphemus is a featherlike structure; its flagellum consists of about 30 stem segments each giving off two pairs of side branches. The antenna develops during the pupal stage (lasting in total about 21 days) from a leaf-shaped anlage by incisions proceeding from the periphery towards the prospective antennal stem. Primary incisions, starting about 3 days after apolysis, form double branches, which arethen split into single branches by parallel running secondary incisions. The initial pattern of tracheae and peripheral nerves is completely rearranged during these morphogenetic processes which are finished 9-10 days after apolysis. In Antheraea the dorsal and ventral epithelial monolayers of the antennal anlage are successively subdivided during development into a pattern of repetitive epithelial zones. Within the first day after apolysis alternating stripes of sensillogenic and non-sensillogenic epithelium are differentiating. Then the latter are further subdivided, and at last four different stripelike zones (I-IV) can be discriminated. Long basal protrusions of the epidermal cells ('epidermal feet'), and most probably haemocytes, seem to be involved in the reconstruction of the epithelium: both show characteristic arrangements within the antennal anlage during successive developmental stages.     

Predictions of Taylor's power law, density dependence and pink noise from a neutrally modeled time series

There has recently been increasing interest in neutral models of biodiversity and their ability to reproduce the patterns observed in nature, such as species abundance distributions. Here we investigate the ability of a neutral model to predict phenomena observed in single-population time series, a study complementary to most existing work that concentrates on snapshots in time of the whole community. We consider tests for density dependence, the dominant frequencies of population fluctuation (spectral density) and a relationship between the mean and variance of a fluctuating population (Taylor's power law). We simulated an archipelago model of a set of interconnected local communities with variable mortality rate, migration rate, speciation rate, size of local community and number of local communities. Our spectral analysis showed ‘pink noise’: a departure from a standard random walk dynamics in favor of the higher frequency fluctuations which is partly consistent with empirical data. We detected density dependence in local community time series but not in metacommunity time series. The slope of the Taylor's power law in the model was similar to the slopes observed in natural populations, but the fit to the power law was worse. Our observations of pink noise and density dependence can be attributed to the presence of an upper limit to community sizes and to the effect of migration which distorts temporal autocorrelation in local time series. We conclude that some of the phenomena observed in natural time series can emerge from neutral processes, as a result of random zero-sum birth, death and migration. This suggests the neutral model would be a parsimonious null model for future studies of time series data.     

Mutational analyses of c-FLIPR, the only murine short FLIP isoform, reveal requirements for DISC recruitment

Cellular FLICE-inhibitory protein (c-FLIP) proteins are known as potent inhibitors of death receptor-mediated apoptosis by interfering with caspase-8 activation at the death-inducing signaling complex (DISC). Among the three human isoforms, c-FLIP(long), c-FLIP(short) and c-FLIP(R), the latter isoform is poorly characterized. We report here the characterization of murine c-FLIP(R) and show that it is the only short c-FLIP isoform expressed in mice. By generating several mutants, we demonstrate that both death effector domains (DEDs) are required for DISC binding and the antiapoptotic function of c-FLIP(R). Surprisingly, the C-terminal tail is important for both protein stability and DISC recruitment. Three-dimensional modeling of c-FLIP(R) revealed a substantial similarity of the overall structures and potential interaction motifs with the viral FLIP MC159. We found, however, that c-FLIP(R) uses different structural motifs for its DISC recruitment. Whereas MC159 interferes with interaction and self-oligomerization of the DISC component FADD by its extensive hydrophilic surface, a narrow hydrophobic patch of c-FLIP(R) on the surface of DED2 is crucial for DISC association. Thus, despite the presence of similar tandem DEDs, viral and cellular FLIPs inhibit apoptosis by remarkably divergent mechanisms.     

Ubiquitin metabolism affects cellular response to volatile anesthetics in yeast

To investigate the mechanism of action of volatile anesthetics, we are studying mutants of the yeast Saccharomyces cerevisiae that have altered sensitivity to isoflurane, a widely used clinical anesthetic. Several lines of evidence from these studies implicate a role for ubiquitin metabolism in cellular response to volatile anesthetics: (i) mutations in the ZZZ1 gene render cells resistant to isoflurane, and the ZZZ1 gene is identical to BUL1 (binds ubiquitin ligase), which appears to be involved in the ubiquitination pathway; (ii) ZZZ4, which we previously found is involved in anesthetic response, is identical to the DOA1/UFD3 gene, which was identified based on altered degradation of ubiquitinated proteins; (iii) analysis of zzz1Delta zzz4Delta double mutants suggests that these genes encode products involved in the same pathway for anesthetic response since the double mutant is no more resistant to anesthetic than either of the single mutant parents; (iv) ubiquitin ligase (MDP1/RSP5) mutants are altered in their response to isoflurane; and (v) mutants with decreased proteasome activity are resistant to isoflurane. The ZZZ1 and MDP1/RSP5 gene products appear to play important roles in determining effective anesthetic dose in yeast since increased levels of either gene increases isoflurane sensitivity whereas decreased activity decreases sensitivity. Like zzz4 strains, zzz1 mutants are resistant to all five volatile anesthetics tested, suggesting there are similarities in the mechanisms of action of a variety of volatile anesthetics in yeast and that ubiquitin metabolism affects response to all the agents examined.