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Mass spectrometry-based "omics" technologies are important tools for global and detailed mapping of post translational modifications. Protein glycosylation is an abundant and important post translational modification widespread throughout all domains of life. Characterization of glycoproteins, including identification of glycan structure and components, their attachment sites and protein carriers, remains challenging. However, recent advances in glycoproteomics, a subbranch that studies and categorizes protein glycosylations, have greatly expanded the known protein glycosylation space and research in this area is rapidly accelerating. Here, we review recent developments in glycoproteomic technologies with a special focus on microbial protein glycosylation.

KCNQ1 is the pore-forming subunit of cardiac slow-delayed rectifier potassium (IKs) channels. Mutations in the kcnq1 gene are the leading cause of congenital long QT syndrome (LQTS). Here, we present the cryoelectron microscopy (cryo-EM) structure of a KCNQ1/calmodulin (CaM) complex. The conformation corresponds to an "uncoupled," PIP2-free state of KCNQ1, with activated voltage sensors and a closed pore. Unique structural features within the S4-S5 linker permit uncoupling of the voltage sensor from the pore in the absence of PIP2. CaM contacts the KCNQ1 voltage sensor through a specific interface involving a residue on CaM that is mutated in a form of inherited LQTS. Using an electrophysiological assay, we find that this mutation on CaM shifts the KCNQ1 voltage-activation curve. This study describes one physiological form of KCNQ1, depolarized voltage sensors with a closed pore in the absence of PIP2, and reveals a regulatory interaction between CaM and KCNQ1 that may explain CaM-mediated LQTS.

The pathophysiology of hidradenitis suppurativa (HS) is not well understood. Some of our knowledge comes from clinical and epidemiological observations, along with studies of the histopathology and immunohistochemistry of affected skin. More recently, cutaneous molecular studies and transcriptomic analyses have provided additional information regarding inflammatory processes. The chronic cutaneous inflammation, systemic symptoms, and associated comorbidities suggest that HS should be classified as an immune-mediated disease, rather than a primary infectious disease. As such, a proposed integrated disease pathway is presented. At a fundamental level, there appears to be a primary abnormality in the pilosebaceous-apocrine unit, which leads to follicular occlusion, perifollicular cyst development that traps commensal microbes, and rupture into the dermis.This can trigger an exaggerated response of the cutaneous innate immune system. Initially this is an acute event, but ongoing intermittent disease activity can lead to recurrent inflammatory nodules and dermal tunnels. Once underway, the cutaneous inflammation is very difficult to turn off, leading to suppurative inflammation in whole anatomic regions.As the disease progresses, we propose that there is recruitment of the systemic immune system perpetuating the chronic cutaneous inflammatory process.There remains much to be done to understand the pathogenesis and immune signature of this challenging disease. (C) 2017 Frontline Medical Communications

Membrane scission is essential for intracellular trafficking. While BAR domain proteins such as endophilin have been reported in dynamin-independent scission of tubular membrane necks, the cutting mechanism has yet to be deciphered. Here, we combine a theoretical model, in vitro, and in vivo experiments revealing how protein scaffolds may cut tubular membranes. We demonstrate that the protein scaffold bound to the underlying tube creates a frictional barrier for lipid diffusion; tube elongation thus builds local membrane tension until the membrane undergoes scission through lysis. We call this mechanism friction-driven scission (FDS). In cells, motors pull tubes, particularly during endocytosis. Through reconstitution, we show that motors not only can pull out and extend protein-scaffolded tubes but also can cut them by FDS. FDS is generic, operating even in the absence of amphipathic helices in the BAR domain, and could in principle apply to any high-friction protein and membrane assembly.

Mendelian susceptibility to mycobacterial disease is a rare syndrome characterized by severe clinical infections usually caused by weakly virulent mycobacterial species such as Bacillus Calmette-Guerin vaccines and environmental nontuberculous mycobacteria or more virulent mycobacteria as mycobacterium tuberculosis. Since 1996, 9 genes including 7 autosomal (STAT1, IFNGR1, IFNGR2, IL12B, IL12RB1, ISG15, and IRF8) and 2 X-linked genes ( NEMO and CYBB) have been identified. Allelic heterogeneity leaded to recognize about 18 genetic diseases with variable clinical phenotypes, but sharing a same physiological mechanism represented by a defect in human IL-12-dependant-INF-gamma-mediated immunity. We report here a case of multifocal Bacillus Calmette-Guerin osteomyelitis in a context Mendelian susceptibility to mycobacterial disease mimicking a metastatic neuroblastoma in a child presenting with delayed growth. The investigation of her twin sister showed the same disease. A heterozygous mutation in exon 22 of STAT1 gene was found in both sisters, another sister and the father being healthy and heterozygous for the same mutation.

1. Community indicators are used to assess the state of ecological communities and to guide management. They are usually calculated from monitoring data, often collected annually. Since any given community indicator provides a univariate summary of complex multivariate phenomena, different changes in the community may lead to the same response in the indicator. Sampling variation can also mask ecologically important trends. 2. This study addresses these challenges for community indicators, with a focus on the large fish indicator (LFI), internationally used to report status of marine fish communities. The LFI expresses 'large' fish biomass as a proportion of total fish biomass and is calculated from species-size-abundance data collected on trawl surveys. We develop new methods to decompose the contributions of species, sampling locations and season to trends over time in the LFI, and highlight consequences for assessment and management. 3. Our results showed that both species and locations made divergent contributions to overall trends in the LFI indicator, with contributions differing by several orders of magnitude and in sign. Only small proportions of species and locations drove overall LFI trends, and their contributions changed with season (spring and autumn surveys). To assess significance of component trends, a resampling method was developed. Our method can be generalized and applied to many other community indicators based on survey data. 4. Synthesis and applications. Our new method for decomposing community indicators and generating confidence intervals makes it possible to extract much more information on what drives a 'headline' indicator, providing a solution to challenges arising from multiple possible interpretations of changes in the indicator and from sampling variation. Analysis of the effects of indicator components on headline indicator values is recommended, because the results allow assessors and managers to identify and interpret how divergent factors (e.g. species, sampling locations and seasons) contribute to the headline indicator value.

The spontaneous emergence of collective flows is a generic property of active fluids and often leads to chaotic flow patterns characterised by swirls, jets, and topological disclinations in their orientation field. However, the ability to achieve structured flows and ordered disclinations is of particular importance in the design and control of active systems. By confining an active nematic fluid within a channel, we find a regular motion of disclinations, in conjunction with a well defined and dynamic vortex lattice. As pairs of moving disclinations travel through the channel, they continually exchange partners producing a dynamic ordered state, reminiscent of Ceilidh dancing. We anticipate that this biomimetic ability to self-assemble organised topological disclinations and dynamically structured flow fields in engineered geometries will pave the road towards establishing new active topological microfluidic devices.

Consciousness can be defined by two major attributes: awareness of environment and self, and arousal, which reflects the level of awareness. The return of arousal after general anesthesia presents an experimental tool for probing the neural mechanisms that control consciousness. Here we have identified that systemic or intracerebral injection of the cannabinoid CB1 receptor (CB1R) antagonist AM281 into the dorsomedial nucleus of the hypothalamus (DMH) - but not the adjacent perifornical area (Pef) or the ventrolateral preoptic nucleus of the hypothalamus (VLPO) - accelerates arousal in mice recovering from general anesthesia. Anesthetics selectively activated endocannabinoid (eCB) signaling at DMH glutamatergic but not GABAergic synapses, leading to suppression of both glutamatergic DMH-Pef and GABAergic DMH-VLPO projections. Deletion of CB1R from widespread cerebral cortical or prefrontal cortical (PFC) glutamatergic neurons, including those innervating the DMH, mimicked the arousal-accelerating effects of AM281. In contrast, CB1R deletion from brain GABAergic neurons or hypothalamic glutamatergic neurons did not affect recovery time from anesthesia. Inactivation of PFC-DMH, DMH-VLPO, or DMH-Pef projections blocked AM281-accelerated arousal, whereas activation of these projections mimicked the effects of AM281. We propose that decreased eCB signaling at glutamatergic terminals of the PFC-DMH projection accelerates arousal from general anesthesia through enhancement of the excitatory DMH-Pef projection, the inhibitory DMH-VLPO projection, or both.

Purpose: To evaluate the effect of catheter-directed irreversible electroporation (iRE) on the integrity, patency, and function of the normal porcine ureter. Materials and Methods: A catheter-mounted electrode was used to perform fluoroscopy-guided IRE in 8 healthy pigs. Two unilateral ablations (90 pulses at 2,000 V, 100 mu s) were performed in each animal in the proximal and distal ureter. Serum creatinine measurements and contrast-enhanced computed tomography imaging were performed at 1, 7, 14, 21, and 28 days after IRE, and findings were compared with baseline values by Student t test. Two animals each were euthanized at 1, 7, 14, and 28 days after IRE for histologic assessment of treatment effects. Quantitative histologic analysis of regeneration and healing of the ureteral wall was graded on a five-point scale. Results: IRE was successfully performed in all animals. Preservation of ureteral wall integrity was confirmed by the leakage-free passage of contrast medium in the treated ureter of all animals through the observation period. Ureteral strictures and associated renal pelvicaliceal dilation were observed in all animals by study days 7 (P = .005) and 14 (P = .007) and did not resolve by day 28. Urothelial recovery was observed in tissue samples from day 7, with progressive replacement of the tunica muscularis with granulation tissue. Despite extensive scarring of the tunica muscularis, full recovery of the urothelium was observed by day 28. Conclusions: The normal porcine ureter retains lumen wall integrity and function following catheter-directed IRE. Scarring of the tunica muscularis in the treated ureter results in stricture formation and reduction of lumen patency.

Using a microfluidic trap, we study the behavior of individual phospholipid vesicles in contact with fatty acids. We show that spontaneous fatty acids insertion inside the bilayer is controlled by the vesicle size, osmotic pressure difference across the membrane and fatty acids concentration in the external bath. Depending on these parameters, vesicles can grow spherically or become unstable and fragment into several daughter vesicles. We establish the phase diagram for vesicle growth and we derive a simple thermodynamic model that reproduces the time evolution of the vesicle volume. Finally, we show that stable growth can be achieved on an artificial cell expressing a simple set of bacterial cytoskeletal proteins, paving the way toward artificial cell reproduction.