Down-regulation of dopamine transporter by iron chelation in vitro is mediated by altered trafficking, not synthesis

Neurological development and functioning of dopamine (DA) neurotransmission is adversely affected by iron deficiency in early life. Iron-deficient rats demonstrate significant elevations in extracellular DA and a reduction in dopamine transporter (DAT) densities in the caudate putamen and nucleus accumbens. To explore possible mechanisms by which cellular iron concentrations control DAT functioning, endogenous DAT-expressing PC12 cells were used to determine the effect of iron chelation on DAT protein and mRNA expression patterns. In addition, we used human DAT (hDAT)-transfected Neuro2a (N2A) cells to examine DAT degradation and trafficking patterns. A 50 microM treatment for 24 h with the iron chelator, desferrioxamine (DFO), significantly decreased dopamine uptake in a dose-dependent manner, with no apparent change in K(m), in both PC12 and N2A cells. Reduced DA uptake was accompanied by concentration- and time-dependent reductions in total DAT protein levels in both cell lines. Exposure to increasing concentrations of DFO did not significantly alter DAT mRNA in either PC12 or N2A cells. However, DAT degradation rates increased three-fivefold in both cell types exposed to 50 microM DFO for 24 h. Biotinylation studies in N2A cells indicate a more dramatic loss of DAT in the membrane fraction, while OptiPrep fractionation experiments revealed an increase in lysosomal DAT with iron chelation. Inhibition of protein kinase C activation with staurosporin prevented the effect of iron chelation on DAT function, suggesting that in vitro iron chelation affects DAT primarily through the effects on trafficking rather than on synthesis.     

Hemin interactions and alterations of the subcellular localization of prion protein

Hemin (iron protoporphyrin IX) is a crucial component of many physiological processes acting either as a prosthetic group or as an intracellular messenger. Some unnatural, synthetic porphyrins have potent anti-scrapie activity and can interact with normal prion protein (PrPC). These observations raised the possibility that hemin, as a natural porphyrin, is a physiological ligand for PrPC. Accordingly, we evaluated PrPC interactions with hemin. When hemin (3-10 microM) was added to the medium of cultured cells, clusters of PrPC formed on the cell surface, and the detergent solubility of PrPC decreased. The addition of hemin also induced PrPC internalization and turnover. The ability of hemin to bind directly to PrPC was demonstrated by hemin-agarose affinity chromatography and UV-visible spectroscopy. Multiple hemin molecules bound primarily to the N-terminal third of PrPC, with reduced binding to PrPC lacking residues 34-94. These hemin-PrPC interactions suggest that PrPC may participate in hemin homeostasis, sensing, and/or uptake and that hemin might affect PrPC functions.     

Bioinformatic analysis of the TonB protein family

TonB is a protein prevalent in a large number of Gram-negative bacteria that is believed to be responsible for the energy transduction component in the import of ferric iron complexes and vitamin B₁₂ across the outer membrane. We have analyzed all the TonB proteins that are currently contained in the Entrez database and have identified nine different clusters based on its conserved 90-residue C-terminal domain amino acid sequence. The vast majority of the proteins contained a single predicted cytoplasmic transmembrane domain; however, nine of the TonB proteins encompass a ∼290 amino acid N-terminal extension homologous to the MecR1 protein, which is composed of three additional predicted transmembrane helices. The periplasmic linker region, which is located between the N-terminal domain and the C-terminal domain, is extremely variable both in length (22–283 amino acids) and in proline content, indicating that a Pro-rich domain is not a required feature for all TonB proteins. The secondary structure of the C-terminal domain is found to be well preserved across all families, with the most variable region being between the second α-helix and the third β-strand of the antiparallel β-sheet. The fourth β-strand found in the solution structure of the Escherichia coli TonB C-terminal domain is not a well conserved feature in TonB proteins in most of the clusters. Interestingly, several of the TonB proteins contained two C-terminal domains in series. This analysis provides a framework for future structure-function studies of TonB, and it draws attention to the unusual features of several TonB proteins. Byron C. H. Chu and R. Sean Peacock contributed equally to this work.     

Disentangling Competition, Herbivory, and Seasonal Effects on Young Plants in Newly Restored Communities

Optimizing techniques of impact and consequence assessment are critical when faced with the challenges of reclamation within a damaged or altered ecosystem. Much debate has arisen over an appropriate index to evaluate herbivore and competition effects on restored communities. We assessed concurrent environmental pressures by means of repeated measurements using three common indices of plant performance (biomass, shoot extension, and survival) in conjunction with monitoring for number and timing of plants eaten. Our design incorporated 24 species, representing a range of taxonomic groups and growth forms, planted at low and high densities, inside and outside large‐scale mammal exclosures. We demonstrate that biomass and height measurements are correlated (at both the individual and the combined species levels), whereas the survival index often showed independent information. Using the most conservative measure (survival), we delineate between plant deaths attributed to seasonal effects, competition (some facilitation was apparent), and herbivory (both compensation and loss of fitness were demonstrated). Plant spacing effects depended on the index (response variable) and whether we measured individual or combined species. The survival index rarely showed competition effects. Due to counter facilitation effects, competition was not demonstrated for any index at the combined species level. The comparison of the relative order and magnitude of plants being eaten against impact identified vulnerable and compensating species. Once identified, compensating species may be used sacrificially to buffer damage in new reclamation systems, whereas deterrents may be used around known vulnerable species.     

Record error and range contraction, real and imagined, in the restricted shrub Banksia hookeriana in south-western Australia

Banksia hookeriana Meissn. (Proteaceae) is a fire‐killed shrub endemic to the northern sandplains of south‐western Australia that could be described as endangered based on its small geographical range (< 5000 km²) and area of occupancy (～500 km²). Impacts on the species’ geographical range by land clearance for farming and mining, and by altered fire regime, were investigated using three lines of evidence: records of herbarium collections, a comprehensive field survey of extant populations, and air photo and satellite images revealing the recent history of land clearance and fires. These show that the species’ range has contracted by up to 40% in area and 26% latitudinally through the loss of outlier and range limit populations since 1960. In addition, 22% of remaining native shrubland on the Eneabba sandplain has been lost over this period through clearing for farming and mining, representing further habitat loss for B. hookeriana. Detailed investigation of B. hookeriana herbarium collections (n = 46) revealed important errors that artificially affected the geographical range of the species and emphasized the importance of close examination of all data captured from collection records. Recorded locations occurred hundreds of kilometres outside the current geographical range of the species in areas with different climate and substrate. Incorrect species identification of herbarium specimens further extended the apparent geographical range of the species. On the other hand, credible records indicated the loss of the species from localities at the limits of its range. Overall, a disconcertingly high proportion of records contained errors that may be difficult to identify without close examination of the original collections and detailed ground‐truthing. Were these records to be used to model climate envelopes, identify potential habitat where the species might occur, or might migrate to either as pests or under climate change scenarios, or to analyse evolutionary or ecological theory (for example) — as is now becoming commonplace — large errors may ensue.     

Plant structural traits and their role in anti-herbivore defence

We consider the role that key structural traits, such as spinescence, pubescence, sclerophylly and raphides, play in protecting plants from herbivore attack. Despite the likelihood that many of these morphological characteristics may have evolved as responses to other environmental stimuli, we show that each provides an important defence against herbivore attack in both terrestrial and aquatic ecosystems. We conclude that leaf-mass–area is a robust index of sclerophylly as a surrogate for more rigorous mechanical properties used in herbivory studies. We also examine herbivore counter-adaptations to plant structural defence and illustrate how herbivore attack can induce the deployment of intensified defensive measures. Although there have been few studies detailing how plant defences vary with age, we show that allocation to structural defences is related to plant ontogeny. Age-related changes in the deployment of structural defences plus a paucity of appropriate studies are two reasons why relationships with other plant fitness characteristics may be obscured, although we describe studies where trade-offs between structural defence and plant growth, reproduction, and chemical defences have been demonstrated. We also show how resource availability influences the expression of structural defences and demonstrate how poorly our understanding of plant structural defence fits into contemporary plant defence theory. Finally, we suggest how a better understanding of plant structural defence, particularly within the context of plant defence syndromes, would not only improve our understanding of plant defence theory, but enable us to predict how plant morphological responses to climate change might influence interactions at the individual (plant growth trade-offs), species (competition), and ecosystem (pollination and herbivory) levels.     

Carpal tunnel syndrome due to an atypical deep soft tissue leiomyoma: The risk of misdiagnosis and mismanagement

Background

Eccrine carcinoma is a quite rare malignant tumor that typically arises from a normal sweat gland and that features a rather high recurrence rate subsequent to simple excision. Given its rather poor response to adjuvant therapy, wide excision of the lesion with tumor-free margins may offer a reasonable chance for long-term control of this neoplasm.

Case presentation

Herein, we report on an unusual case of perianal eccrine carcinoma, initially presenting as a perianal abscess.

Conclusion

Even though eccrine carcinomas would appear to be rare, when dealing with recurrent skin tumors or recurrent perianal fistulas, the possibility of eccrine carcinoma should be considered by consulting clinicians.