Air-Sea Exchange and Atmospheric Deposition of Phthalate Esters in the South China Sea.

Phthalate esters (PAEs) have been investigated in paired air and seawater samples collected onboard the research vessel SONNE in the South China Sea in the summer of 2019. The concentrations of ∑7PAEs ranged from 2.84 to 24.3 ng/m3 with a mean of 9.67 ± 5.86 ng/m3 in air and from 0.96 to 8.35 ng/L with a mean of 3.05 ng/L in seawater. Net air-to-seawater deposition dominated air-sea exchange fluxes of DiBP, DnBP, DMP, and DEP, while strong water-to-air volatilization was estimated for bis(2-ethylhexyl) phthalate (DEHP). The estimated net atmospheric depositions were 3740 t/y for the sum of DMP, DEP, DiBP, and DnBP, but DEHP volatilized from seawater to air with an average of 900 t/y. The seasonally changing monsoon circulation, currents, and cyclones occurring in the Pacific can significantly influence the concentration of PAEs, and alter the direction and magnitude of air-sea exchange and particle deposition fluxes. Consequently, the dynamic air-sea exchange process may drive the transport of PAEs from marginal seas and estuaries toward remote marine environments, which can play an important role in the environmental transport and cycling of PAEs in the global ocean.

Benthic ecosystem functioning under climate change: modelling the bioturbation potential for benthic key species in the southern North Sea.

Climate change affects the marine environment on many levels with profound consequences for numerous biological, chemical, and physical processes. Benthic bioturbation is one of the most relevant and significant processes for benthic-pelagic coupling and biogeochemical fluxes in marine sediments, such as the uptake, transport, and remineralisation of organic carbon. However, only little is known about how climate change affects the distribution and intensity of benthic bioturbation of a shallow temperate shelf sea system such as the southern North Sea. In this study, we modelled and projected changes in bioturbation potential (BPp) under a continuous global warming scenario for seven southern North Sea key bioturbators: Abra alba, Amphiura filiformis, Callianassa subterranea, Echinocardium cordatum, Goniada maculata, Nephtys hombergii, and Nucula nitidosa. Spatial changes in species bioturbation intensity are simulated for the years 2050 and 2099 based on one species distribution model per species driven by bottom temperature and salinity changes using the IPCC SRES scenario A1B. Local mean bottom temperature was projected to increase between 0.15 and 5.4 °C, while mean bottom salinity was projected to moderately decrease by 1.7. Our results show that the considered benthic species are strongly influenced by the temperature increase. Although the total BP remained rather constant in the southern North Sea, the BPp for four out of seven species was projected to increase, mainly due to a simultaneous northward range expansion, while the BPp in the core area of the southern North Sea declined for the same species. Bioturbation of the most important species, Amphiura filiformis and Echinocardium cordatum, showed no substantial change in the spatial distribution, but over time. The BPp of E. cordatum remained almost constant until 2099, while the BPp of A. filiformis decreased by 41%. The northward expansion of some species and the decline of most species in the south led to a change of relative contribution to bioturbation in the southern North Sea. These results indicate that some of the selected key bioturbators in the southern North Sea might partly compensate the decrease in bioturbation by others. But especially in the depositional areas where bioturbation plays a specifically important role for ecosystem functioning, bioturbation potential declined until 2099, which might affect the biochemical cycling in sediments of some areas of the southern North Sea.

Occurrence and distribution of estrogenic substances in the northern South China Sea.

Estrogenic substances are today among the contaminants of emerging concern. Besides naturally occurring estrogens, other natural and synthetic substances can mimic a hormonal action due to their structural resemblance to hormones, possibly affecting the endocrine system of living organisms. Estrogens have been detected in inland water bodies such as influents and effluents of waste water treatment plants as well as in rivers, but data on their distribution and variability in the marine ecosystem are still limited. Surface water samples obtained during two research cruises on the northern shelf of the South China Sea (SCS) near the Pearl River Estuary, in September 2018 and in August 2019, were investigated for estrogenic substances, namely estrone (E1), 17ß-estradiol (E2), 17α-ethinylestradiol (EE2), genistein (GEN), daidzein (DAI) and zearalenone (ZEN). Among the target analytes, the natural hormones E1 and E2, as well as the synthetic EE2, were the most abundant with maximum concentrations of 1.1 ng L-1, 0.7 ng L-1 and 0.6 ng L-1, respectively. Of substances produced by plants and fungi, GEN, DAI and ZEN, only GEN was detected (1.2 ng L-1). High concentrations occurred predominantly close to the coast, which was also reflected in the calculated estradiol equivalent quotients (up to 1.4 ng L-1). In general, the distribution of estrogenic substances observed in both years shows a regional and inter-annual variability consistent with the modeled surface current data for the SCS. Regarding single estrogenic compounds and estradiol equivalents, marine organisms in the northern SCS might be exposed to high potential risk.

Climate change effects on marine protected areas: Projected decline of benthic species in the North Sea.

Climate change is a global threat for marine ecosystems, their biodiversity and consequently ecosystem services. In the marine realm, marine protected areas (MPAs) were designated to counteract regional pressures, but they might be ineffective to protect vulnerable species and habitats, if their distribution is affected by global climate change. We used six Species Distribution Models (GLM, MARS, FDA, RF, GBM, MAXENT) to project changes in the distribution of eight benthic indicator and key species under climate change in the North Sea MPAs for 2050 and 2099. The projected distribution area of most species will be stable or even increase within the MPAs between 2001 and 2050. Thereafter, the distribution area decreased, especially within MPAs in the central North Sea by 2099, and some key species even disappeared from the MPAs. Consequently, the monitoring and protection of benthic species might not be possible within static MPA borders under climate change.

The multi PAM2 protein Upa2 functions as novel core component of endosomal mRNA transport.

mRNA transport determines spatiotemporal protein expression. Transport units are higher-order ribonucleoprotein complexes containing cargo mRNAs, RNA-binding proteins and accessory proteins. Endosomal mRNA transport in fungal hyphae belongs to the best-studied translocation mechanisms. Although several factors are known, additional core components are missing. Here, we describe the 232 kDa protein Upa2 containing multiple PAM2 motifs (poly[A]-binding protein [Pab1]-associated motif 2) as a novel core component. Loss of Upa2 disturbs transport of cargo mRNAs and associated Pab1. Upa2 is present on almost all transport endosomes in an mRNA-dependent manner. Surprisingly, all four PAM2 motifs are dispensable for function during unipolar hyphal growth. Instead, Upa2 harbours a novel N-terminal effector domain as important functional determinant as well as a C-terminal GWW motif for specific endosomal localisation. In essence, Upa2 meets all the criteria of a novel core component of endosomal mRNA transport and appears to carry out crucial scaffolding functions.

Core components of endosomal mRNA transport are evolutionarily conserved in fungi.

Active movement of mRNAs by sophisticated transport machineries determines precise spatiotemporal expression of encoded proteins. A prominent example discovered in fungi is microtubule-dependent transport via endosomes. This mode of transport was thought to be only operational in the basidiomycete Ustilago maydis. Here, we report that distinct core components are evolutionarily conserved in fungal species of distantly related phyla like Mucoromycota. Interestingly, orthologues of the key RNA-binding protein Rrm4 from the higher basidiomycete Coprinopsis cinerea and the mucoromycete Rhizophagus irregularis shuttle on endosomes in hyphae of U. maydis. Thus, endosomal mRNA transport appears to be more wide-spread than initially anticipated.

mRNA transport in fungal top models.

Eukaryotic cells rely on the precise determination of when and where proteins are synthesized. Spatiotemporal expression is supported by localization of mRNAs to specific subcellular sites and their subsequent local translation. This holds true for somatic cells as well as for oocytes and embryos. Most commonly, mRNA localization is achieved by active transport of the molecules along the actin or microtubule cytoskeleton. Key factors are molecular motors, adaptors, and RNA-binding proteins that recognize defined sequences or structures in cargo mRNAs. A deep understanding of this process has been gained from research on fungal model systems such as Saccharomyces cerevisiae and Ustilago maydis. Recent highlights of these studies are the following: (1) synergistic binding of two RNA-binding proteins is needed for high affinity recognition; (2) RNA sequences undergo profound structural rearrangements upon recognition; (3) mRNA transport is tightly linked to membrane trafficking; (4) mRNAs and ribosomes are transported on the cytoplasmic surface of endosomes; and (5) heteromeric protein complexes are, most likely, assembled co-translationally during endosomal transport. Thus, the study of simple fungal model organisms provides valuable insights into fundamental mechanisms of mRNA transport boosting the understanding of similar events in higher eukaryotes. WIREs RNA 2018, 9:e1453. doi: 10.1002/wrna.1453 This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein-RNA Recognition RNA Interactions with Proteins and Other Molecules > RNA-Protein Complexes RNA Export and Localization > RNA Localization.

The ESCRT regulator Did2 maintains the balance between long-distance endosomal transport and endocytic trafficking.

In highly polarised cells, like fungal hyphae, early endosomes function in both endocytosis as well as long-distance transport of various cargo including mRNA and protein complexes. However, knowledge on the crosstalk between these seemingly different trafficking processes is scarce. Here, we demonstrate that the ESCRT regulator Did2 coordinates endosomal transport in fungal hyphae of Ustilago maydis. Loss of Did2 results in defective vacuolar targeting, less processive long-distance transport and abnormal shuttling of early endosomes. Importantly, the late endosomal protein Rab7 and vacuolar protease Prc1 exhibit increased shuttling on these aberrant endosomes suggesting defects in endosomal maturation and identity. Consistently, molecular motors fail to attach efficiently explaining the disturbed processive movement. Furthermore, the endosomal mRNP linker protein Upa1 is hardly present on endosomes resulting in defects in long-distance mRNA transport. In conclusion, the ESCRT regulator Did2 coordinates precise maturation of endosomes and thus provides the correct membrane identity for efficient endosomal long-distance transport.

The mitochondrial alternative oxidase Aox1 is needed to cope with respiratory stress but dispensable for pathogenic development in Ustilago maydis.

The mitochondrial alternative oxidase is an important enzyme that allows respiratory activity and the functioning of the Krebs cycle upon disturbance of the respiration chain. It works as a security valve in transferring excessive electrons to oxygen, thereby preventing potential damage by the generation of harmful radicals. A clear biological function, besides the stress response, has so far convincingly only been shown for plants that use the alternative oxidase to generate heat to distribute volatiles. In fungi it was described that the alternative oxidase is needed for pathogenicity. Here, we investigate expression and function of the alternative oxidase at different stages of the life cycle of the corn pathogen Ustilago maydis (Aox1). Interestingly, expression of Aox1 is specifically induced during the stationary phase suggesting a role at high cell density when nutrients become limiting. Studying deletion strains as well as overexpressing strains revealed that Aox1 is dispensable for normal growth, for cell morphology, for response to temperature stress as well as for filamentous growth and plant pathogenicity. However, during conditions eliciting respiratory stress yeast-like growth as well as hyphal growth is strongly affected. We conclude that Aox1 is dispensable for the normal biology of the fungus but specifically needed to cope with respiratory stress.

A FYVE zinc finger domain protein specifically links mRNA transport to endosome trafficking.

An emerging theme in cellular logistics is the close connection between mRNA and membrane trafficking. A prominent example is the microtubule-dependent transport of mRNAs and associated ribosomes on endosomes. This coordinated process is crucial for correct septin filamentation and efficient growth of polarised cells, such as fungal hyphae. Despite detailed knowledge on the key RNA-binding protein and the molecular motors involved, it is unclear how mRNAs are connected to membranes during transport. Here, we identify a novel factor containing a FYVE zinc finger domain for interaction with endosomal lipids and a new PAM2-like domain required for interaction with the MLLE domain of the key RNA-binding protein. Consistently, loss of this FYVE domain protein leads to specific defects in mRNA, ribosome, and septin transport without affecting general functions of endosomes or their movement. Hence, this is the first endosomal component specific for mRNP trafficking uncovering a new mechanism to couple mRNPs to endosomes.

The Vip1 inositol polyphosphate kinase family regulates polarized growth and modulates the microtubule cytoskeleton in fungi.

Microtubules (MTs) are pivotal for numerous eukaryotic processes ranging from cellular morphogenesis, chromosome segregation to intracellular transport. Execution of these tasks requires intricate regulation of MT dynamics. Here, we identify a new regulator of the Schizosaccharomyces pombe MT cytoskeleton: Asp1, a member of the highly conserved Vip1 inositol polyphosphate kinase family. Inositol pyrophosphates generated by Asp1 modulate MT dynamic parameters independent of the central +TIP EB1 and in a dose-dependent and cellular-context-dependent manner. Importantly, our analysis of the in vitro kinase activities of various S. pombe Asp1 variants demonstrated that the C-terminal phosphatase-like domain of the dual domain Vip1 protein negatively affects the inositol pyrophosphate output of the N-terminal kinase domain. These data suggest that the former domain has phosphatase activity. Remarkably, Vip1 regulation of the MT cytoskeleton is a conserved feature, as Vip1-like proteins of the filamentous ascomycete Aspergillus nidulans and the distantly related pathogenic basidiomycete Ustilago maydis also affect the MT cytoskeleton in these organisms. Consistent with the role of interphase MTs in growth zone selection/maintenance, all 3 fungal systems show aspects of aberrant cell morphogenesis. Thus, for the first time we have identified a conserved biological process for inositol pyrophosphates.

Kinesin-3 and dynein mediate microtubule-dependent co-transport of mRNPs and endosomes.

Long-distance transport of mRNAs is important in determining polarity in eukaryotes. Molecular motors shuttle large ribonucleoprotein complexes (mRNPs) containing RNA-binding proteins and associated factors along microtubules. However, precise mechanisms including the interplay of molecular motors and a potential connection to membrane trafficking remain elusive. Here, we solve the motor composition of transported mRNPs containing the RNA-binding protein Rrm4 of the pathogen Ustilago maydis. The underlying transport process determines the axis of polarity in infectious filaments. Plus-end-directed Kin3, a kinesin-3 type motor, mediates anterograde transport of mRNPs and is also present in transport units moving retrogradely. Split dynein Dyn1-Dyn2 functions in retrograde movement of mRNPs. Plus-end-directed conventional kinesin Kin1 is indirectly involved by transporting minus-end-directed dynein back to plus ends. Importantly, we additionally demonstrate that Rrm4-containing mRNPs colocalise with the t-SNARE Yup1 on shuttling endosomes and that functional endosomes are essential for mRNP movement. Either loss of Kin3 or removal of its lipid-binding pleckstrin-homology domain abolishes Rrm4-dependent movement without preventing colocalisation of Rrm4 and Yup1-positive endosomes. In summary, we uncovered the combination of motors required for mRNP shuttling along microtubules. Furthermore, intimately linked co-transport of endosomes and mRNPs suggests vesicle hitchhiking as mode of mRNP transport.

Fungal development of the plant pathogen Ustilago maydis.

The maize pathogen Ustilago maydis has to undergo various morphological transitions for the completion of its sexual life cycle. For example, haploid cells respond to pheromone by forming conjugation tubes that fuse at their tips. The resulting dikaryon grows filamentously, expanding rapidly at the apex and inserting retraction septa at the basal pole. In this review, we present progress on the underlying mechanisms regulating such defined developmental programmes. The key findings of the postgenomic era are as follows: (1) endosomes function not only during receptor recycling, but also as multifunctional transport platforms; (2) a new transcriptional master regulator for pathogenicity is part of an intricate transcriptional network; (3) determinants for uniparental mitochondrial inheritance are encoded at the a2 mating-type locus; (4) microtubule-dependent mRNA transport is important in determining the axis of polarity; and (5) a battery of fungal effectors encoded in gene clusters is crucial for plant infection. Importantly, most processes are tightly controlled at the transcriptional, post-transcriptional and post-translational levels, resulting in a complex regulatory network. This intricate system is crucial for the timing of the correct order of developmental phases. Thus, new insights from all layers of regulation have substantially advanced our understanding of fungal development.

Dissolved oxygen and its response to eutrophication in a tropical black water river.

The Siak is a typical, nutrient-poor, well-mixed, black water river in central Sumatra, Indonesia, which owes its brown color to dissolved organic matter (DOM) leached from surrounding, heavily disturbed peat soils. We measured dissolved organic carbon (DOC) and oxygen concentrations along the river, carried out a 36-h experiment in the province capital Pekanbaru and quantified organic matter and nutrient inputs from urban wastewater channels into the Siak. In order to consider the complex dynamic of oxygen in rivers, a box-diffusion model was used to interpret the measured data. The results suggest that the decomposition of soil derived DOM was the main factor influencing the oxygen concentration in the Siak which varied between approximately 100 and 140 micromol l(-1). Additional DOM input caused by wastewater discharges appeared to reduce the oxygen concentrations by approximately 20 micromol l(-1) during the peak-time in household water use in the early morning and in the early evening. Associated enhanced nutrient inputs appear to reduce the impact of the anthropogenic DOM by favoring the photosynthetic production of oxygen in the morning. A reduction of 20 micromol l(-1), which although perhaps not of great significance in Pekanbaru, has strong implications for wastewater management in the fast developing areas downstream Pekanbaru where oxygen concentrations rarely exceed 20 micromol l(-1).

The fungal RNA-binding protein Rrm4 mediates long-distance transport of ubi1 and rho3 mRNAs.

Cytoskeletal transport promotes polar growth in filamentous fungi. In Ustilago maydis, the RNA-binding protein Rrm4 shuttles along microtubules and is crucial for polarity in infectious filaments. Mutations in the RNA-binding domain cause loss of function. However, it was unclear which RNAs are bound and transported. Here, we applied in vivo RNA binding studies and live imaging to determine the molecular function of Rrm4. This new combination revealed that Rrm4 mediates microtubule-dependent transport of distinct mRNAs encoding, for example, the ubiquitin fusion protein Ubi1 and the small G protein Rho3. These transcripts accumulate in ribonucleoprotein particles (mRNPs) that move bidirectionally along microtubules and co-localise with Rrm4. Importantly, the 3' untranslated region of ubi1 containing a CA-rich binding site functions as zipcode during mRNA transport. Furthermore, motile mRNPs are not formed when the RNA-binding domain of Rrm4 is deleted, although the protein is still shuttling. Thus, Rrm4 constitutes an integral component of the transport machinery. We propose that microtubule-dependent mRNP trafficking is crucial for hyphal growth introducing U. maydis as attractive model for studying mRNA transport in higher eukaryotes.

Mass budgets and contribution of individual sources and sinks to the abundance of gamma-HCH, alpha-HCH and PCB 153 in the North Sea.

Mass budgets of hexachlorocyclohexanes (alpha-HCH and gamma-HCH) and a polychlorinated biphenyl (PCB 153) for 1995--2001 were calculated based on model simulations and observations for the North Sea as a whole and the German Bight, a coastal shallow subregion. For the North Sea the air-sea fluxes of the three pollutants were net depositional and dominated by local sources (gamma-HCH and PCB 153) or atmospheric deposition (alpha-HCH). The air-sea fluxes were net volatilizational in the German Bight. Unlike HCH, PCB 153 does not show a downward trend in the North Sea marine environment during the study period. Due to its physicochemical properties it is expected to readily enter the food chains. Model results suggest that during studied period, the North Sea was a sink for PCB 153 and a source of HCHs for the outer world.

Thermodynamic and structural equivalence of two HLA-B27 subtypes complexed with a self-peptide.

The F pocket of major histocompatibility complex (in humans HLA) class I molecules accommodates the C terminus of the bound peptide. Residues forming this pocket exhibit considerable polymorphism, and a single difference (Asp116 in HLA-B*2705 and His116 in HLA-B*2709 heavy chains) confers differential association of these two HLA-B27 subtypes to the autoimmune disease ankylosing spondylitis. As peptide presentation by HLA molecules is of central importance for immune responses, we performed thermodynamic (circular dichroism, differential scanning calorimetry, fluorescence polarization) and X-ray crystallographic analyses of both HLA-B27 subtypes complexed with the epidermal growth factor response factor 1-derived self-peptide TIS (RRLPIFSRL) to understand the impact of the Asp116His exchange on peptide display. This peptide is known to be presented in vivo by both subtypes, and as expected for a self-peptide, TIS-reactive cytotoxic T lymphocytes are absent in the respective individuals. The thermodynamic analyses reveal that both HLA-B27:TIS complexes exhibit comparable, relatively high thermostability (Tm approximately 60 degrees C) and undergo multi-step unfolding reactions, with dissociation of the peptide in the first step. As shown by X-ray crystallography, only subtle structural differences between the subtypes were observed regarding the architecture of their F pockets, including the presence of distinct networks of water molecules. However, no consistent structural differences were found between the peptide presentation modes. In contrast to other peptides displayed by the two HLA-subtypes which show either structural or dynamical differences in their peptide presentation modes, the TIS-complexed HLA-B*2705 and HLA-B*2709 subtypes are an example for thermodynamic and structural equivalence, in agreement with functional data.

Differential peptide dynamics is linked to major histocompatibility complex polymorphism.

Peptide presentation by major histocompatibility complex (MHC) molecules is of central importance for immune responses, which are triggered through recognition of peptide-loaded MHC molecules (pMHC) by cellular ligands such as T-cell receptors (TCR). However, a unifying link between structural features of pMHC and cellular responses has not been established. Instead, pMHC/TCR binding studies suggest conformational and/or flexibility changes of the binding partners as a possible cause of differential T-cell stimulation, but information on real-time dynamics is lacking. We therefore probed the real-time dynamics of a MHC-bound nonapeptide (m9), by combining time-resolved fluorescence depolarization and molecular dynamics simulations. Here we show that the nanosecond dynamics of this peptide presented by two human MHC class I subtypes (HLA-B*2705 and HLA-B*2709) with differential autoimmune disease association varies dramatically, despite virtually identical crystal structures. The peptide dynamics is linked to the single, buried polymorphic residue 116 in the peptide binding groove. Pronounced peptide flexibility is seen only for the non-disease-associated subtype HLA-B*2709, suggesting an entropic control of peptide recognition. Thermodynamic data obtained for two additional peptides support this hypothesis.

Rheopheresis in patients with ischemic diabetic foot syndrome: results of an open label prospective pilot trial.

Rheopheresis is a specific application of membrane differential filtration, synonymous with double filtration plasmapheresis, for extracorporeal hemorheotherapy. Safety and efficacy of Rheopheresis for wound healing and skin oxygenation were investigated in patients with ischemic diabetic foot syndrome. Eight patients with type 2 diabetes mellitus and non-healing foot ulcers caused by severe ischemic diabetic foot syndrome were treated by a series of seven Rheopheresis sessions in a time span of 11 weeks. Wound healing had not been detectable under conditions of standardized wound care during at least 2 months. Wound status was classified by its morphology, severity and location, according to the criteria of Wagner. Transcutaneous oxygen pressure (tcPO2), laser Doppler flowmetry and vital capillary microscopy were repeatedly performed to monitor the effects of the Rheopheresis treatment series on microcirculation and skin blood flow. Laboratory parameters of blood rheology, endothelial function and inflammatory state were measured in addition to safety parameters. In four patients (baseline Wagner stage 2), Rheopheresis accelerated wound healing of foot ulcers and was associated with an improvement of Wagner stage and a pronounced increase in tcPO2. In two patients (baseline Wagner stage 2), wound healing was unchanged but mean tcPO2 increased, allowing successful minor amputation. Values of tcPO2 remained stable and enhanced for the 3 months follow-up period. In two patients (baseline Wagner stage 4 or 5), no improvements in foot lesions were observed within the treatment period. As an adjunct therapeutic option, Rheopheresis may preserve a functional lower extremity, delay amputation or reduce the extent of amputation.