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In contrast to the widely used sequence stratigraphic models for passive continental margins, the stacking patterns of strata within epeiric seas, which are influenced by regional tectonic activity, may display opposing characteristics during the same geological period. These variations serve as a record of basin evolution and also affect the accumulation of hydrocarbons within the strata. Our study investigated the development potential of the deep Longmaxi Shale in the southern Sichuan Basin by examining the sequence stratigraphy and sedimentary fill patterns. Using a combination of core observation, well-logging data analysis, and 3D seismic profile interpretation, we aimed to gain an understanding of the sedimentary fill history of the Longmaxi Shale during the Early Silurian. Our analysis revealed that deglaciation and regional tectonic events affected the sequence stratigraphy, resulting in unconformities that were identifiable using seismic data and wireline logs. Through an analysis of thirty wireline logs and two seismic profiles, we identified two third-order sequences suggested in the Lower Longmaxi Formation. Within the two third-order sequences were five systems tracts, with the first exhibiting a complete cycle of sea-level change and the second cycle being incomplete due to regional tectonic events. The graptolite succession on the upper Yangtze Platform provided a temporal view of the sequence stratigraphy and sedimentation rates of the Longmaxi Shale. The thickness trends of the systems tracts reflected the interplay of short-term eustasy fluctuations, subsidence, and uplift. Our analysis suggests that regional subsidence played a significant role in the deposition of the second transgressive systems tract (TST) in the Weiyuan and Luzhou areas, which represents a promising target for shale gas exploration, in addition to the first TST. However, the Changning area experienced a relative sea-level decrease due to the intense uplift of the Qianzhogn Paleo-uplift and the increased supply of sediment and is interpreted as a highstand systems tract (HST); it is not considered to have shale gas exploration potential.

Atopic dermatitis (AD) is a common, chronic-relapsing inflammatory skin disease with significant disease burden. Genetic and environmental trigger factors contribute to AD, activating 2 of our largest organs, the nervous system and the immune system. Dysregulation of neuroimmune circuits plays a key role in the pathophysiology of AD, causing inflammation, pruritus, pain, and barrier dysfunction. Sensory nerves can be activated by environmental or endogenous trigger factors, transmitting itch stimuli to the brain. On stimulation, sensory nerve endings also release neuromediators into the skin, contributing again to inflammation, barrier dysfunction, and itch. In addition, dysfunctional peripheral and central neuronal structures contribute to neuroinflammation, sensitization, nerve elongation, and neuropathic itch, thus chronification and therapy resistance. Consequently, neuroimmune circuits in skin and central nervous system may be targets to treat pruritus in AD. Cytokines, chemokines, proteases, lipids, opioids, and ions excite/sensitize sensory nerve endings, which not only induces itch but further aggravates/perpetuates inflammation, skin barrier disruption, and pruritus as well. Thus, targeted therapies for neuroimmune circuits as well as pathway inhibitors (eg, kinase inhibitors) may be beneficial to control pruritus in AD either in systemic and/or in topical form. Understanding neuroimmune circuits and neuronal signaling will optimize our approach to control all pathological mechanisms in AD, inflammation, barrier dysfunction, and pruritus.

The clinical significance and regulators of IL-13Rα2 in itch and atopic dermatitis (AD) remain unclear. To identify disease-driven regulatory circuits of IL-13Rα2, transcriptomic/pathological analysis was performed in skin from patients with AD, psoriasis, healthy subjects, and murine AD model. Functionality was investigated in sensory neurons, keratinocytes and animal model, by using knockdown (KD), calcium imaging, RNA-seq, cytokine arrays, pharmacological assays, and behavioural investigations. In our study, an upregulated IL-13Rα2 expression was revealed in skin of AD patients, but not psoriasis, in a disease activity-dependent manner. In cultured human keratinocytes, IL-13 increased IL-13Rα2 transcription levels, and this were downregulated by IL-13Rα1KD. IL-13Rα2KD reduced transcription levels of EDNRA, CCL20, CCL26. In contrast, sensory neuron-derived IL-13Rα2 was upregulated by TLR2 heterodimer agonists, Pam3CSK4 and FSL-1. In a mouse cheek model, pre-administration of Pam3CSK4 and FSL-1 enhanced IL-13-elicited scratching behaviour. Consistently, in cultured sensory neurons Pam3CSK4 enhanced IL-13-elicted calcium transients, increased number of responders, and orchestrated chemerin, CCL17 and CCL22 release. These release was inhibited by IL-13Rα2KD. Collectively, IL-13 regulates keratinocyte-derived IL-13Rα2 and TLR2 to modulate neuronal IL-13Rα2, thereby promoting neurogenic inflammation and exacerbating AD and itch. Thus, the cutaneous IL-13-IL-13Rα2 and neuronal TLR2-IL-13Rα2 pathway represent important targets to treat AD and itch.

The Upper Ordovician Wufeng Formation and the Lower Silurian Longmaxi Formation in the central Yangtze area of southern China has a good prospect of shale gas exploration. So far, however, its complicated geological conditions and shale gas exploration and development potentials have not been understood completely, which affects its exploration achievements. In order to determine the main factors controlling shale gas enrichment in the Wufeng–Longmaxi Fms in this area, this paper studied the key shale gas enrichment conditions in the black shale there (e.g. sedimentary environments, reservoir development characteristics and preservation conditions) based on single-well comprehensive reservoir evaluation, combined with drilling, logging, core, outcrop and test data. In addition, it was compared with the main shale gas blocks in the Sichuan Basin, such as Jiaoshiba and Changning. And the following research results are obtained. First, the organic-rich shale section in this area was formed during the sedimentation from Wufeng Formation to the third submember of the first member of Longmaxi Formation. It is the deposit of deepwater continental facies, and its thickness is in a range of 15–39 m, presenting a trend of increasing from south to north. Second, the reservoir rocks in the high-quality shale sections are dominated by siliceous shale, and the reservoir porosity is in a range of 1.60–7.44%. The reservoir spaces are dominated by organic pores with good connectivity and high total organic carbon (TOC) content. The organic matter is of a sapropel–sapropel prone hybrid type, with a high thermal evolution degree, better gas bearing property and good fracturability. Third, the Dangyang synclinorium is characterized by better strata preservation, rock occurrence flat, less developed faults and thick and stable roofs and floors, so its shale gas preservation conditions are better. Fourth, different from the Changning and Jiaoshiba Blocks, the central Yangtze area is characterized by great burial depth, large bidirectional stress difference, and low formation pressure coefficient. In conclusion, the Wufeng–Longmaxi Fms is better in shale gas enrichment conditions and has a potential of further exploration. However, its commercial shale gas development and scale production increase in the future face challenges due to its large stress difference, great burial depth, and lower pressure coefficient.

Atopic dermatitis (AD) is a chronic skin disease, which is associated with intense itch, skin barrier dysfunction and eczematous lesions. Aberrant IL-20 expression has been implicated in numerous inflammatory diseases, including psoriasis. However, the role of IL-20 in AD remains unknown. Here, RNA-seq, Q-PCR, and immunocytochemistry were utilized to examine disease-driven changes of IL-20 and its cognate receptor subunits in skin from healthy human subjects, AD patients and murine AD-models. Calcium imaging, knockdown and cytokine array were used to investigate IL-20-evoked responses in keratinocytes and sensory neurons. The murine cheek model and behavioral scoring were employed to evaluate IL-20-elicited sensations in vivo. We found that transcripts and protein of IL-20 were upregulated in skin from human AD and murine AD-like models. Topical MC903 treatment in mice ear enhanced IL-20R1 expression in the trigeminal sensory ganglia, suggesting a lesion-associated and epidermal-driven mechanism for sensitization of sensory IL-20 signaling. IL-20 triggered calcium influx in both keratinocytes and sensory neurons, and promoted their AD-related molecule release and transcription of itch-related genes. In sensory neurons, IL-20 application increased TLR2 transcripts, implicating a link between innate immune response and IL-20. In a murine cheek model of acute itch, intradermal injection IL-20 and IL-13 elicited significant itch-like behavior, though only when co-injected. Our findings provide novel insights into IL-20 function in peripheral (skin-derived) itch and clinically relevant intercellular neuron-epidermal communication, highlighting a role of IL-20 signaling in the pathophysiology of AD, thus forming a new basis for the development of a novel antipruritic strategy via interrupting IL-20 epidermal pathways.

Chronic itch is a complex sensation of the skin frequently associated with skin diseases, such as atopic dermatitis (AD) and psoriasis. Although Serpin E1 is implicated in chronic itch, its receptor and signaling pathways involved in itch are not known. In this study, the clinical relevance of a putative Serpin E1 receptor PLAUR to chronic itch, and the neuro-cutaneous Serpin E1-PLAUR signaling are explored. We found that PLAUR is overexpressed in skin specimens of human lesional AD and lesional psoriasis, and sensory neurons innervating MC903-induced AD-like murine skin. Murine PLAUR

Chronic itch poses major health care and economic burdens worldwide. In 2013, B‐type natriuretic peptide (BNP) was identified as an itch‐selective neuropeptide and shown to be both necessary and sufficient to produce itch behaviour in mice. Since then, mechanistic studies of itch have increased, not only at central levels of the spinal relay of itch signalling but also in the periphery and skin. In this review, we have critically analysed recent findings from complementary pharmacological and physiological approaches, combined with genetic strategies to examine the role of BNP in itch transduction and modulation of other pruritic proteins. Additionally, potential targets and possible strategies against BNP signalling are discussed for developing novel therapeutics in itch. Overall, we aim to provide insights into drug development by altering BNP signalling to modulate disease symptoms in chronic itch, including conditions for which no approved treatment exists.

Chronic pain is a leading health and socioeconomic problem and an unmet need exists for long-lasting analgesics. SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) are required for neuropeptide release and noxious signal transducer surface trafficking, thus, selective expression of the SNARE-cleaving light-chain protease of botulinum neurotoxin A (LCA) in peripheral sensory neurons could alleviate chronic pain. However, a safety concern to this approach is the lack of a sensory neuronal promoter to prevent the expression of LCA in the central nervous system. Towards this, we exploit the unique characteristics of Pirt (phosphoinositide-interacting regulator of TRP), which is expressed in peripheral nociceptive neurons. For the first time, we identified a Pirt promoter element and cloned it into a lentiviral vector driving transgene expression selectively in peripheral sensory neurons. Pirt promoter driven-LCA expression yielded rapid and concentration-dependent cleavage of SNAP-25 in cultured sensory neurons. Moreover, the transcripts of pain-related genes (TAC1, tachykinin precursor 1, &nbsp, CALCB, calcitonin gene-related peptide 2, HTR3A, 5-hydroxytryptamine receptor 3A, NPY2R, neuropeptide Y receptor Y2, GPR52, G protein-coupled receptor 52, SCN9A, sodium voltage-gated channel alpha subunit 9, TRPV1 and TRPA1, transient receptor potential cation channel subfamily V member 1 and subfamily A member 1) in pro-inflammatory cytokines stimulated sensory neurons were downregulated by viral mediated expression of LCA. Furthermore, viral expression of LCA yielded long-lasting inhibition of pain mediator release. Thus, we show that the engineered Pirt-LCA virus may provide a novel means for long lasting pain relief.

Paleogene lacustrine dolomicrite of Unit II of the lower Xingouzui Formation in the southern part of the Qianjiang Depression, China forms “tight carbonate” hydrocarbon reservoirs that can be divided into the following four dolomicrite types: dolomicrite, muddy dolomicrite, sandy dolomicrite, and gypsiferous dolomicrite. These four dolomicrite types occur in the following three distinct combinations, which are referred to as subfacies: (1) interbedded dolomicrite (or sandy dolomicrite) and dark-gray siliciclastic mudstone, interpreted as open lacustrine deposits, (2) grayish-dark siliciclastic mudstone with thinly-bedded muddy dolomicrite or gypsiferous dolomicrite, interpreted as restricted lacustrine deposits, and (3) grayish-green mudstone with thinlybedded brown-yellow siltstone, interpreted as lacustrine shoreline deposits. The spatial distribution of these subfacies was controlled by the paleo-provenance system, paleo-microgeomorphology, and lake level fluctuations. In the open lacustrine subfacies, dolomicrite or sandy dolomicrite are often vertically interbedded with dark argillite rocks, forming a suitable petroleum source rockreservoir association. The dolomicrite and sandy dolomicrite reservoirs possess many secondary crystalline pores and few dissolution pores, with mesopore porosities in the range of 15%–25%. The dark mud shale interval has total organic carbon (TOC) values exceeding 4% (averaging 1.29%). The organic matter is mainly type II2, with vitrinite reflectance values averaging 0.57, which indicates potential for the generation of immature oil. The oil-source correlation results indicated that the n-alkanes of the crude oil and source rock samples from Unit II of the lower Xingouzui Formation exhibit single peak values, with a main peak (post-peak) in C22 and a Pr/Ph ratio $#x003C; 1 in a C2920R $#x003E; C2720R $#x003E; C2820R distribution. These features indicate that source-reservoir interbedding is characteristic of selfsourced reservoirs. The interbedded dolomicrite (or sandy dolomicrite) and dark-gray siliciclastic mudstone, i.e., subfacies (1), is a favorable target for hydrocarbons in the study area.

Transient receptor potential (TRP) vallinoid 1 (TRPV1) and ankyrin 1 (TRPA1) are two transducing channels expressed on peripheral sensory nerves involved in pain sensation. Upregulation of their expression, stimulated by inflammatory cytokines and growth factors in animal pain models, correlate with the induction of nociceptive hyper-sensitivity. Herein, we firstly demonstrate by immuno-cytochemical labelling that TNFα augments the surface content of these channels on rat cultured dorsal root ganglion (DRG) neurons which, in turn, enhances the electrophysiological and functional responses of the latter to their specific agonists. A molecular basis underlying this TNFα-dependent enhancement was unveiled by pre-treating DRGs with a recently-published chimeric protein, consisting of the protease light chain (LC) of botulinum neurotoxin (BoNT) serotype E fused to full-length BoNT/A (LC/E-BoNT/A). This cleaves synaptosomal-associated protein of Mr 25k (SNAP-25) and reported previously to exhibit anti-nociceptive activity in a rat model of neuropathic pain. Low pM concentrations of this chimera were found to prevent the TNFα-stimulated delivery of TRPV1/A1 to the neuronal plasmalemma and, accordingly, decreased their incremental functional activities relative to those of control cells, an effect accompanied by SNAP-25 cleavage. Advantageously, LC/E-BoNT/A did not reduce the basal surface contents of the two channels or their pharmacological responses. Thus, use of multiple complementary methodologies provides evidence that LC/E-BoNT/A abolishes the TNFα-dependent augmented, but not resting, surface trafficking of TRPV1/A1. As TNFα is known to induce nociceptive hyper-sensitivity in vivo, our observed inhibition by LC/E-BoNT/A of its action in vitro could contribute to its potential alleviation of pain.

It has been disputed whether volcanic activity promotes the enrichment of organic matter in geological periods. In a biostratigraphic framework, this study divided the volcanic eruption periods during the Ordovician-Silurian transition in western Hubei Province based on the longitudinal differentiation of the bentonite development, analyzed the sedimentological and geochemical characteristics of each eruption period, and explored the relationship between the volcanic activity and the organic matter enrichment. The results show that from early to late, the Ordovician-Silurian transition can be divided into 1) a strong eruption period (lower Wufeng) that was mainly dominated by carbonaceous shale deposits (containing both terrigenous silicon and biological silicon, with a medium organic matter content), which corresponded to a stage of strong flexural subsidence; 2) a weak eruption period (Wufeng-Longmaxi transition) that was dominated by organic-rich and biogenic silica-rich siliceous shale deposits, which corresponded to a stage of stable flexural subsidence; 3) a strong eruption period (lower Longmaxi) that was dominated by carbonaceous shale deposits with a medium organic matter content, which involved major changes in the paleogeographic pattern and corresponded to an initial stage of flexural migration wherein the depositional water body became shallow; and 4) a strong eruption period (upper Longmaxi) dominated by organic-poor, terrigenous silica-rich argillaceous shale deposits, which corresponded to a stage of expanded flexural migration wherein the depositional water body became even shallower. The strength of the volcanic activity was closely associated with the differential enrichment of the organic matter. The tectonic activity was stable in the period of weak volcanic activity, which was characterized by low terrigenous inputs, moderate water restriction, and a high paleo-productivity. This was favorable for organic matter enrichment, thereby leading to a high TOC content. During the periods of violent volcanic activity, the Cathaysia Block collided violently with the Yangtze blocks, resulting in more terrigenous input, a higher degree of water restriction, a shallower depositional water body, and a lower paleo-productivity, which had a significant inhibitory effect on the organic matter enrichment. This inhibitory effect was larger than the promotional effect of the volcanic ash on the organic matter enrichment, thereby resulting in a decrease in the TOC content.

A pressing need exists for long-acting, non-addictive medicines to treat chronic pain, a major societal burden. Botulinum neurotoxin type A (BoNT/A) complex – a potent, specific and prolonged inhibitor of neuro-exocytosis – gives some relief in several pain disorders, but not for all patients. Our study objective was to modify BoNT/A to overcome its inability to block transmitter release elicited by high [Ca 2+ ] i and increase its limited analgesic effects. This was achieved by fusing a BoNT/A gene to that for the light chain (LC) of type/E. The resultant purified protein, LC/E-BoNT/A, entered cultured sensory neurons and, unlike BoNT/A, inhibited release of calcitonin gene-related peptide evoked by capsaicin. Western blotting revealed that this improvement could be due to a more extensive truncation by LC/E of synaptosomal-associated protein of Mr = 25 k, essential for neuro-exocytosis. When tested in a rat spared nerve injury (SNI) model, a single intra-plantar (IPL) injection of LC/E-BoNT/A alleviated for ∼2 weeks mechanical and cold hyper-sensitivities, in a dose-dependent manner. The highest non-paralytic dose (75 U/Kg, IPL) proved significantly more efficacious than BoNT/A (15 U/Kg, IPL) or repeated systemic pregabalin (10 mg/Kg, intraperitoneal), a clinically-used pain modulator. Effects of repeated or delayed injections of this fusion protein highlighted its analgesic potential. Attenuation of mechanical hyperalgesia was extended by a second administration when the effect of the first had diminished. When injected 5 weeks after injury, LC/E-BoNT/A also reversed fully-established mechanical and cold hyper-sensitivity. Thus, combining advantageous features of BoNT/E and/A yields an efficacious, locally-applied and long-acting anti-hyperalgesic.

The role of soluble N-ethylmaleimide-sensitive factor attachment protein receptors in atopic dermatitis (AD) is unknown. This study identifies the function of soluble N-ethylmaleimide sensitive factor attachment protein receptor in AD-related cytokine secretion and epidermis-nerve communication. Herein, we report that various cytokines were simultaneously upregulated and coreleased in innate immunity–activated primary human keratinocytes. AD-related cytokines thymic stromal lymphopoietin, endothelin-1, and inflammatory tumor necrosis factor-α activated distinct but overlapping sensory neurons. Tumor necrosis factor-α potentiated thymic stromal lymphopoietin–induced Ca2+-influx, whereas endothelin-1 caused itch-selective B-type natriuretic peptide release. In primary human keratinocytes, B-type natriuretic peptide upregulated genes promoting dermatological and neuroinflammatory diseases and conditions. VAMP3, SNAP-29, and syntaxin 4 proved important in driving cytokine release from primary human keratinocytes. Depletion of VAMP3 inhibited nearly all the cytokine release including thymic stromal lymphopoietin and endothelin-1. Accordingly, VAMP3 co-occurred with endothelin-1 in the skins of patients with AD. Our study pinpoints the pivotal role of soluble N-ethylmaleimide sensitive factor attachment protein receptors in mediating cytokine secretion related to AD. VAMP3 is identified as a suitable target for developing broad-spectrum anticytokine therapeutics for controlling itch and atopic skin inflammation.

This study discovered a novel neuro-epidermal BNP-TRPV3-Serpin E1-mediated pathway in severe atopic dermatitis (AD). Serpin E1 was identified as a new itch-inducer. We propose this pathway represents an important target for the treatment of AD.

The Cambrian Jiulaodong Formation of the Wei-201 well block in the Sichuan Basin was investigated for shale gas potential. In the subsurface, the thermally mature formation attained a stable thickness of 234 m encompassing an area of approximately [Formula: see text] and representing a potential gas resource. The total gas content measurements from canistered samples was more than the estimated total gas storage capacity of the free gas, absorbed gas, and gas dissolved in water and in oil. The canister gas content ranged between 0.971 and [Formula: see text] and averaged [Formula: see text]. The average estimated gas in place was 2.5 billion cubic meters for the formation in the Weiyuan area. Reflectance measurements for thermal maturity range between 2.60% and 3.06% and average 2.84%. The results of our total organic carbon content (TOC) content analysis conducted on the core shale samples indicate that the TOC content of the formation ranges from 0.87% to 3.57% and averages 2.2%. The mineral composition of marine mudstone formation of the Jiulaodong shale is relatively consistent. Brittle mineral content increases with organic carbon content and is approximately 32%–43%, of which quartz content is 29%–40% with a very low amount of clay mineral as the mixed layer. The amount of illite-smectite ranges from 0% to 1% and the brittleness index range from 37% to 62% and average 57.1%. The Cambrian Jiulaodong Formation ha very good petroleum-source rock potential due to its average TOC content of greater than 2%, average canister gas content of [Formula: see text], good type I kerogen, high maturity with average 2.84% of source rocks that are characterized by a fairly high abundance of organic matter increasing from top to bottom and a large thickness of 234 m. Natural fractures, cracks, and pores developed in the Jiulaodong Formation also provide space for shale gas storage, and its average brittleness index is greater than 57%, which is good for fracability.

The function of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) in cellular trafficking, membrane fusion and vesicle release in synaptic nerve terminals is well characterised. Recent studies suggest that SNAREs are also important in the control of tumourigenesis through the regulation of multiple signalling and transportation pathways. The majority of published studies investigated the effects of knockdown/knockout or overexpression of particular SNAREs on the normal function of cells as well as their dysfunction in tumourigenesis promotion. SNAREs are involved in the regulation of cancer cell invasion, chemo-resistance, the transportation of autocrine and paracrine factors, autophagy, apoptosis and the phosphorylation of kinases essential for cancer cell biogenesis. This evidence highlights SNAREs as potential targets for novel cancer therapy. This is the first review to summarise the expression and role of SNAREs in cancer biology at the cellular level, their interaction with non-SNARE proteins and modulation of cellular signalling cascades. Finally, a strategy is proposed for developing novel anti-cancer therapeutics using targeted delivery of a SNARE-inactivating protease into malignant cells.

Background T H 2 cell–released IL-31 is a critical mediator in patients with atopic dermatitis (AD), a prevalent and debilitating chronic skin disorder. Brain-derived natriuretic peptide (BNP) has been described as a central itch mediator. The importance of BNP in peripheral (skin-derived) itch and its functional link to IL-31 within the neuroimmune axis of the skin is unknown. Objective We sought to investigate the function of BNP in the peripheral sensory system and skin in IL-31–induced itch and neuroepidermal communication in patients with AD. Methods Ca 2+ imaging, immunohistochemistry, quantitative real-time PCR, RNA sequencing, knockdown, cytokine/phosphokinase arrays, enzyme immune assay, and pharmacologic inhibition were performed to examine the cellular basis of the IL-31–stimulated, BNP-related itch signaling in dorsal root ganglionic neurons (DRGs) and skin cells, transgenic AD-like mouse models, and human skin of patients with AD and healthy subjects. Results In human DRGs we confirmed expression and co-occurrence of oncostatin M receptor β subunit and IL-31 receptor A in a small subset of the neuronal population. Furthermore, IL-31 activated approximately 50% of endothelin-1–responsive neurons, and half of the latter also responded to histamine. In murine DRGs IL-31 upregulated Nppb and induced soluble N-ethylmaleimide–sensitive factor activating protein receptor–dependent BNP release. In Grhl3PAR2 /+ mice house dust mite–induced severe AD-like dermatitis was associated with Nppb upregulation. Lesional IL-31 transgenic mice also exhibited increased Nppb transcripts in DRGs and the skin; accordingly, skin BNP receptor levels were increased. Importantly, expression of BNP and its receptor were increased in the skin of patients with AD. In human skin cells BNP stimulated a proinflammatory and itch-promoting phenotype. Conclusion For the first time, our findings show that BNP is implicated in AD and that IL-31 regulates BNP in both DRGs and the skin. IL-31 enhances BNP release and synthesis and orchestrates cytokine and chemokine release from skin cells, thereby coordinating the signaling pathways involved in itch. Inhibiting peripheral BNP function might be a novel therapeutic strategy for AD and pruritic conditions.

Botulinum neurotoxins (BoNTs) are the most toxic proteins known, due to inhibiting the neuronal release of acetylcholine and causing flaccid paralysis. Most BoNT serotypes target neurons by binding to synaptic vesicle proteins and gangliosides via a C-terminal binding sub-domain (HCC). However, the role of their conserved N-terminal sub-domain (HCN) has not been established. Herein, we created a mutant form of recombinant BoNT/A lacking HCN (rAΔHCN) and showed that the lethality of this mutant is reduced 3.3 × 104-fold compared to wild-type BoNT/A. Accordingly, low concentrations of rAΔHCN failed to bind either synaptic vesicle protein 2C or neurons, unlike the high-affinity neuronal binding obtained with 125I-BoNT/A (Kd = 0.46 nM). At a higher concentration, rAΔHCN did bind to cultured sensory neurons and cluster on the surface, even after 24 h exposure. In contrast, BoNT/A became internalised and its light chain appeared associated with the plasmalemma, and partially co-localised with vesicle-associated membrane protein 2 in some vesicular compartments. We further found that a point mutation (W985L) within HCN reduced the toxicity over 10-fold, while this mutant maintained the same level of binding to neurons as wild type BoNT/A, suggesting that HCN makes additional contributions to productive internalization/translocation steps beyond binding to neurons.

The Boxing Sag is located in the southwest of Dongying Depression, southern Bohai Bay Basin of East China. It is one of the main petroliferous sags in the Dongying Depression, and has two major source rocks, namely the upper 4th Member (Es4u) and lower 3rd Member (Es3l) shales of Eocene Shahejie Formation, which are the new exploration targets in recent years. In this study, 16 core samples were collected from Es4u and Es3l shales in the Boxing Sag, and the saturate hydrocarbons were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS), respectively. The results show that Es4u shale has obvious phytane and gammacerane predominance, higher concentration of tricyclic terpanes and regular steranes, and very low concentration of 4-methly steranes; Es3l shale has pristane predominance, lower concentration of tricyclic terpanes, gammacerane and regular steranes, and higher concentration of 4-methly steranes. The Es4u shale can be further divided into two types based on the distribution of n-alkanes in gas chromatograms: normal distribution and double peak pattern. The biomarker characteristics and sedimentary facies distribution show that Es4u shale was deposited in the saline-hypersaline semi-deep (Type A Es4u shale, sag center) to shallow (Type B Es4u shale, sag edge) lacustrine environments, Es3l shale was deposited in the freshwater-brackish semi-deep-deep lacustrine environments, and the former sedimentary facies maps of Es4u and Es3l in the Boxing Sag are further modified.

Oligomerisation of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes is required for synaptic vesicle fusion and neurotransmitter release. How these regulate the release of pain peptides elicited by different stimuli from sensory neurons has not been established. Herein, K(+) depolarization was found to induce multiple sodium dodecyl sulfate (SDS)-resistant SNARE complexes in sensory neurons exposed to botulinum neurotoxins (BoNTs), with molecular weights ranging from 104-288 k (large) to 38-104 k (small). Isoform 1 of vesicle-associated membrane protein 1 (VAMP 1) assembled into stable complexes upon depolarisation and was required for the participation of intact synaptosome-associated protein of relative molecular mass 25 k (SNAP-25) or BoNT/A-truncated form (SNAP-25A) in the large functional and small inactive SDS-resistant SNARE complexes. Cleaving VAMP 1 decreased SNAP-25A in the functional complexes to a much greater extent than the remaining intact SNAP-25. Syntaxin 1 proved essential for the incorporation of intact and SNAP-25A into the large complexes. Truncation of syntaxin 1 by BoNT/C1 caused /A- and/or /C1-truncated SNAP-25 to appear in non-functional complexes and blocked the release of calcitonin gene-related peptide (CGRP) elicited by capsaicin, ionomycin, thapsigargin or K(+) depolarization. Only the latter two were susceptible to /A. Inhibition of CGRP release by BoNT/A was reversed by capsaicin and/or ionomycin, an effect overcome by BoNT/C1. Unlike BoNT/B, BoNT/D cleaved VAMP 1 in addition to 2 and 3 in rat sensory neurons and blocked both CGRP and substance P release. Thus, unlike SNAP-25, syntaxin 1 and VAMP 1 are more suitable targets to abolish functional SNARE complexes and pain peptide release evoked by any stimuli.

Proteins responsible for basal and stimulated endocytosis in nerves containing small clear synaptic vesicles (SCSVs) or large dense-core vesicles (LDCVs) are revealed herein, using probes that exploit surface-exposed vesicle proteins as acceptors for internalization. Basal uptake of botulinum neurotoxins (BoNTs) by both SCSV-releasing cerebellar granule neurons (CGNs) and LDCV-enriched trigeminal ganglionic neurons (TGNs) was found to require protein acceptors and acidic compartments. In addition, dynamin, clathrin, adaptor protein complex-2 (AP2), and amphiphysin contribute to the depolarization-evoked entry. For fast recycling of SCSVs, knockdown and knockout strategies demonstrated that CGNs use predominantly dynamin 1, whereas isoform 2 and, to a smaller extent, isoform 3 support a less rapid mode of stimulated endocytosis. Accordingly, proximity ligation assay confirmed that dynamin 1 and 2 colocalize with amphiphysin 1 in CGNs, and the latter copurified with both dynamins from cell extracts. In contrast, LDCV-releasing TGNs preferentially employ dynamins 2 and 3 and amphiphysin 1 for evoked endocytosis and lack the fast phase. Hence, stimulation recruits dynamin, clathrin, AP2, and amphiphysin to augment BoNT internalization, and neurons match endocytosis mediators to the different demands for locally recycling SCSVs or replenishing distally synthesized LDCVs.

The Paleogene “Red Bed”, the alternative strata for oil and gas exploration in the South Slope of the Dongying Depression, refers to a set of red clastic sediments of the lower 4th member of the Shahejie Formation and the 1st member of the Kongdian Formation. And it has important significance to study the controlling factors on oil and gas accumulation and the accumulation modes for guiding the deep-layer oil and gas exploration in the area. In this paper, according to the statistics of oil and gas static characteristics, analysis of oil original types, study of pathway system and recovery of oil and gas accumulation, the controlling factors on the Red Bed oil and gas accumulation were summarized as follows: source rock controlling oil and gas distribution, physical features of reservoir bed controlling trap oiliness, faults controlling oil and gas source and trap conditions, and nose structural zone controlling oil and gas migration and enrichment zone. Based on these studies, three accumulation modes for the Paleogene Red Bed in the research area were built: joints of source rock and reservoir bed acting as lateral pathway for oil and gas accumulation, faults acting as vertical pathway for oil and gas accumulation, and high pressure of deeper depression resulting in oil and gas flowing downward to form accumulation.

// Jianghui Meng 1,2 1 Charles Institute of Dermatology, School of Medicine and Medical Sciences, University College Dublin, Belfield, Dublin, Ireland 2 International Centre for Neurotherapeutics, Dublin City University, Glasnevin, Dublin, Ireland Correspondence to: Jianghui Meng, email: // email: // Keywords : endocytosis, tumor, cancer target, clathrin, amphiphysin Received : September 28, 2016 Accepted : March 09, 2017 Published : March 29, 2017 Abstract Dynamins and their related proteins participate in the regulation of neurotransmission, antigen presentation, receptor internalization, growth factor signalling, nutrient uptake, and pathogen infection. Recently, emerging findings have shown dynamin proteins can also contribute to the genesis of cancer. This up-to-date review herein focuses on the functionality of dynamin in cancer development. Dynamin 1 and 2 both enhance cancer cell proliferation, tumor invasion and metastasis, whereas dynamin 3 has tumor suppression role. Antisense RNAs encoded on the DNA strand opposite a dynamin gene regulate the function of dynamin, and manipulate oncogenes and tumor suppressor genes. Certain dynamin-related proteins are also upregulated in distinct cancer conditions, resulting in apoptotic resistance, cell migration and poor prognosis. Altogether, dynamins are potential biomarkers as well as representing promising novel therapeutic targets for cancer treatment. This study also summarizes the current available dynamin-targeted therapeutics and suggests the potential strategy based on signalling pathways involved, providing important information to aid the future development of novel cancer therapeutics by targeting these dynamin family members.

Transient receptor potential (TRP) A1 and V1 channels relay sensory signals, yet little is known about their transport to the plasmalemma during inflammation. Herein, TRPA1 and TRPV1 were found on vesicles containing calcitonin gene-related peptide (CGRP), accumulated at sites of exo- and endo-cytosis and co-localised on fibres and cell bodies of cultured sensory neurons expressing both. A proinflammatory cytokine, TNFα, elevated their surface content and both resided in close proximity, indicating co-trafficking. Syntaxin 1–interacting protein, Munc18–1, proved necessary for the response to TNFα and for TRPV1-triggered CGRP release. TNFα-induced surface trafficking of TRPV1 and TRPA1 required a synaptic vesicle membrane protein VAMP1 (but not 2/3), which is essential for CGRP exocytosis from large dense-core vesicles. Inactivation of two proteins on the presynaptic plasma membrane, syntaxin-1 or SNAP-25, by botulinum neurotoxin (BoNT)/C1 or /A inhibited the TNFα-elevated delivery. Accordingly, enhancement by TNFα of Ca2+ influx through the upregulated surface-expressed TRPV1 and TRPA1 channels was abolished by BoNT/A. Thus, in addition, the neurotoxins’ known inhibition of the release of pain transmitters, their therapeutic potential is augmented by lowering the exocytotic delivery of transducing channels and the resultant hyper-sensitisation in inflammation.