Pulpal upregulation of connexin 43 during pulpitis.

OBJECTIVES: Connexins are building blocks of membranous channels that form gap junctions and hemichannels. These channels are essential portals for information exchange and coordination during inflammation. Pathologic levels of these conduits may result in excessive inflammation and collateral destruction. This study aimed to analyse temporospatial levels of connexin 43 (Cx43) during pulpitis in extracted human teeth and in a rodent model. A specific interest was directed at the pulpal stroma as it is conserved during vital pulp therapy. MATERIALS AND METHODS: Pulpal tissues were attained from human extracted teeth of various pulpal inflammatory stages and fixed for cryosections. Pulpal exposures were created in bilateral maxillary molars in Sprague-Dawley rats. Rats were sacrificed at days 1 to 5 post-exposure. Immunofluorescence histology was performed to detect Cx43, markers for inflammation, and cell death. Immunofluorescent levels in the pulpal stroma at 3 sites (wound/near/far) were matched to pulpal condition (human) or days post-exposure (rodent). RESULTS: Cx43 upregulation was observed with increased severity of pulpitis both in humans and rodent model. The upregulation appeared to be global and included distant regions. Elevated levels of neutrophils were present in advanced pulpitis. Apoptosis and necroptosis seem to be upregulated in human samples as Cx43 levels rose. CONCLUSIONS: We observed a disseminated upregulation of Cx43 throughout the pulpal stroma as inflammation became advanced. This observation may facilitate cell death signal transfer or represent overt levels of purinergic signalling that leads to pro-inflammatory conditions. CLINICAL RELEVANCE: Cx43 downregulation may represent a potential therapeutic approach to enable resolution of pulpal inflammation.

Interpretation and application of Sponge City guidelines in China.

'Sponge City' is the term used to describe the Chinese government's approach to urban surface water management. The concept was conceived in 2014 in response to an increasing incidence of urban flooding or water-logging in Chinese cities. While ambitious and far-reaching in its aim (of decreasing national flood risk, increasing water supply and improving water quality), the initiative must be implemented by individual subprovincial or municipal-level government entities. Thus, while the concept is similar to sustainable drainage systems (SuDS) in the UK (or low-impact development (LID) in the USA), it is developing with different regional characteristics, and during continuing rapid urbanization. Indeed, the increasing use of national rather than international examples of best practice reflects a growing body of knowledge that has evolved since the start of the Sponge City initiative. In this paper, interpretation and development of the national Sponge City guidelines are assessed for the Ningbo Municipality, an affluent and rapidly expanding city on China's low-lying east coast. While climate, geology and socio-economic factors can all be seen to influence the way that national guidelines are implemented, project financing, integration and assessment are found to be of increasing influence. This article is part of the theme issue 'Urban flood resilience'.

The impact of ischemia-reperfusion injuries on skin resident murine dendritic cells.

Pressure ulcers are a chronic problem for patients or the elderly who require extended periods of bed rest. The formation of ulcers is due to repeated cycles of ischemia-reperfusion (IR), which initiates an inflammatory response. Advanced ulcers disrupt the skin barrier, resulting in further complications. To date, the immunological aspect of skin IR has been understudied, partly due to the complexity of the skin immune cells. Through a combination of mass cytometry, confocal imaging and intravital multiphoton imaging, this study establishes a workflow for multidimensionality single cell analysis of skin myeloid cell responses in the context of IR injury with high spatiotemporal resolution. The data generated has provided us with previously uncharacterized insights into the distinct cellular behavior of resident dendritic cells (DCs) and recruited neutrophils post IR. Of interest, we observed a drop in DDC numbers in the IR region, which was subsequently replenished 48h post IR. More importantly, in these cells, we observe an attenuated response to repeated injuries, which may have implications in the subsequent wound healing process.

Enterococcus faecalis Modulates Immune Activation and Slows Healing During Wound Infection.

Enterococcus faecalis is one of the most frequently isolated bacterial species in wounds yet little is known about its pathogenic mechanisms in this setting. Here, we used a mouse wound excisional model to characterize the infection dynamics of E faecalis and show that infected wounds result in 2 different states depending on the initial inoculum. Low-dose inocula were associated with short-term, low-titer colonization whereas high-dose inocula were associated with acute bacterial replication and long-term persistence. High-dose infection and persistence were also associated with immune cell infiltration, despite suppression of some inflammatory cytokines and delayed wound healing. During high-dose infection, the multiple peptide resistance factor, which is involved in resisting immune clearance, contributes to E faecalis fitness. These results comprehensively describe a mouse model for investigating E faecalis wound infection determinants, and suggest that both immune modulation and resistance contribute to persistent, nonhealing wounds.

Reach-scale variation surface water quality in a reticular canal system in the lower Yangtze River Delta region, China.

The aim of this research was to assess the spatial and temporal distribution of surface water pollution within a reticular canal system typical of those found in the lower Yangtze River Delta (YRD). For this purpose, surface water quality data was collected from a drainage canal that bisected the southeast district of Ningbo Municipality (Zhejiang) from 2013 to 2015. The sampling transect was designed to represent the change in land-use from the agriculture dominated rural hinterland, to the predominantly urban city-centre. To calculate the representative land-use fraction of each sampling location, the contributing area was defined using an uni-directional 1 km vector line-buffer around the 'upstream' section of canal. The spatial and temporal variation of EC, DO, NH3 and turbidity indicated a measureable difference between the urban and rural sections of the channel. Water quality indicators were most sensitive to urban and parkland land-use types. The study yielded an increased spatial resolution to knowledge of water-quality variability in the urban environment compared to previous studies within the YRD region. The results were used to make recommendations for the development of an effective long-term strategy for the improvement in surface water quality in this region.

Contribution of T-type calcium channel isoforms to cold and mechanical sensitivity in naïve and oxaliplatin-treated mice of both sexes.

BACKGROUND AND PURPOSE: The chemotherapy agent oxaliplatin can give rise to oxaliplatin-induced peripheral neuropathy (OIPN). Here, we investigated whether T-type calcium channels (Cav3) contribute to OIPN. EXPERIMENTAL APPROACH: We chronically treated mice with oxaliplatin and assessed pain responses and changes in expression of Cav3.2 calcium channels. We also tested the effects of T-type channel blockers on cold sensitivity in wild-type and Cav3.2 null mice. KEY RESULTS: Oxaliplatin treatment led to mechanical and cold hypersensitivity in male and female mice. Mechanical hypersensitivity persisted in Cav3.2 null mice of both sexes. Intraperitoneal or intrathecal delivery of pan T-type channel inhibitors attenuated mechanical hypersensitivity in wild-type but not Cav3.2 null mice. Remarkably cold hypersensitivity occurred in female but not male Cav3.2 null mice even without oxaliplatin treatment. Unexpectedly, intrathecal, intraplantar or intraperitoneal delivery of T-type channel inhibitors Z944 or TTA-P2 transiently induced cold hypersensitivity in both male and female wild-type mice. Acute knockdown of specific Cav3 isoforms revealed that the depletion of Cav3.1 in males and depletion of either Cav3.1 or Cav3.2 in females triggered cold hypersensitivity. Finally, reducing Cav3.2 expression by disrupting the interactions between Cav3.2 and the deubiquitinase USP5 with the small organic molecule II-2 reversed oxaliplatin-induced mechanical and cold hypersensitivity and importantly did not trigger cold allodynia. CONCLUSION AND IMPLICATIONS: Altogether, our data indicate that T-type channels differentially contribute to the regulation of cold and mechanical hypersensitivity, and raise the possibility that T-type channel blockers could promote cold allodynia.

Mechano-Activated Cell Therapy for Accelerated Diabetic Wound Healing.

Chronic diabetic wounds are a significant global healthcare challenge. Current strategies, such as biomaterials, cell therapies, and medical devices, however, only target a few pathological features and have limited efficacy. A powerful platform technology combining magneto-responsive hydrogel, cells, and wireless magneto-induced dynamic mechanical stimulation (MDMS) is developed to accelerate diabetic wound healing. The hydrogel encapsulates U.S. Food and Drug Administration (FDA)-approved fibroblasts and keratinocytes to achieve ∼3-fold better wound closure in a diabetic mouse model. MDMS acts as a nongenetic mechano-rheostat to activate fibroblasts, resulting in ∼240% better proliferation, ∼220% more collagen deposition, and improved keratinocyte paracrine profiles via the Ras/MEK/ERK pathway to boost angiogenesis. The magneto-responsive property also enables on-demand insulin release for spatiotemporal glucose regulation through increasing network deformation and interstitial flow. By mining scRNAseq data, a mechanosensitive fibroblast subpopulation is identified that can be mechanically tuned for enhanced proliferation and collagen production, maximizing therapeutic impact. The "all-in-one" system addresses major pathological factors associated with diabetic wounds in a single platform, with potential applications for other challenging wound types.

A long-term dataset of topography and nearshore bathymetry at the macrotidal pocket beach of Porsmilin, France.

Long-term datasets documenting the evolution of coastal forms and processes, through the provision of recurring beach as well as shoreface morphological observations and accompanying time-series of environmental controls, remain difficult to collect and are rarely made available. However, they are increasingly needed to further our understanding of coastal change and to improve the models that will help planning what our future coast will be. This data descriptor presents the results of topographic and bathymetric surveys at Porsmilin, a macrotidal embayed beach situated in Brittany, northwest France. The Porsmilin beach survey program was launched in January 2003 by the Institut Universitaire Européen de la Mer (IUEM/Univ. Brest) and is continuing today in the framework of the French coastal observation service SNO-DYNALIT. The dataset contains over 16 years of monthly beach profile surveys and a large collection of repeated high-resolution subtidal and subaerial digital elevation models (DEMs). The dataset is accompanied by time-series of inshore waves and water levels, and enriched metadata, that will facilitate its future reuse in coastal research.

A flexible multiplexed immunosensor for point-of-care in situ wound monitoring.

Chronic wounds arise from interruption of normal healing due to many potential pathophysiological factors. Monitoring these multivariate factors can provide personalized diagnostic information for wound management, but current sensing technologies use complex laboratory tests or track a limited number of wound parameters. We report a flexible biosensing platform for multiplexed profiling of the wound microenvironment, inflammation, and infection state at the point of care. This platform integrates a sensor array for measuring inflammatory mediators [tumor necrosis factor-α, interleukin-6 (IL-6), IL-8, and transforming growth factor-ß1], microbial burden (Staphylococcus aureus), and physicochemical parameters (temperature and pH) with a microfluidic wound exudate collector and flexible electronics for wireless, smartphone-based data readout. We demonstrate in situ multiplexed monitoring in a mouse wound model and also profile wound exudates from patients with venous leg ulcers. This technology may facilitate more timely and personalized wound management to improve chronic wound healing outcomes.

A wireless and battery-free wound infection sensor based on DNA hydrogel.

The confluence of wireless technology and biosensors offers the possibility to detect and manage medical conditions outside of clinical settings. Wound infections represent a major clinical challenge in which timely detection is critical for effective interventions, but this is currently hindered by the lack of a monitoring technology that can interface with wounds, detect pathogenic bacteria, and wirelessly transmit data. Here, we report a flexible, wireless, and battery-free sensor that provides smartphone-based detection of wound infection using a bacteria-responsive DNA hydrogel. The engineered DNA hydrogels respond selectively to deoxyribonucleases associated with pathogenic bacteria through tunable dielectric changes, which can be wirelessly detected using near-field communication. In a mouse acute wound model, we demonstrate that the wireless sensor can detect physiologically relevant amounts of Staphylococcus aureus even before visible manifestation of infection. These results demonstrate strategies for continuous infection monitoring, which may facilitate improved management of surgical or chronic wounds.

A Cav3.2 T-type calcium channel point mutation has splice-variant-specific effects on function and segregates with seizure expression in a polygenic rat model of absence epilepsy.

Low-voltage-activated, or T-type, calcium (Ca(2+)) channels are believed to play an essential role in the generation of absence seizures in the idiopathic generalized epilepsies (IGEs). We describe a homozygous, missense, single nucleotide (G to C) mutation in the Ca(v)3.2 T-type Ca(2+) channel gene (Cacna1h) in the genetic absence epilepsy rats from Strasbourg (GAERS) model of IGE. The GAERS Ca(v)3.2 mutation (gcm) produces an arginine to proline (R1584P) substitution in exon 24 of Cacna1h, encoding a portion of the III-IV linker region in Ca(v)3.2. gcm segregates codominantly with the number of seizures and time in seizure activity in progeny of an F1 intercross. We have further identified two major thalamic Cacna1h splice variants, either with or without exon 25. gcm introduced into the splice variants acts "epistatically," requiring the presence of exon 25 to produce significantly faster recovery from channel inactivation and greater charge transference during high-frequency bursts. This gain-of-function mutation, the first reported in the GAERS polygenic animal model, has a novel mechanism of action, being dependent on exonic splicing for its functional consequences to be expressed.

Major ion chemistry and dissolved inorganic carbon cycling in a human-disturbed mountainous river (the Luodingjiang River) of the Zhujiang (Pearl River), China.

Major ion chemistry and dissolved inorganic carbon system (DIC, mainly HCO3(-) and gaseous CO2) in the Luodingjiang River, a mountainous tributary of the Zhujiang (Pearl River), China, were examined based on a seasonal and spatial sampling scheme in 2005. The diverse distribution of lithology and anthropogenic impacts in the river basin provided the basic idea to assess the effects of lithology vs. human activities on water chemistry and carbon biogeochemistry in river systems. Major ions showed great spatial variations, with higher concentrations of total dissolved solids (TDS) and DIC in the regions with carbonate rocks and clastic sedimentary rocks, while lower in the regions with metamorphic sandstones and schists as well as granites. pCO2 at all sampling sites was oversaturated in June, ranging with a factor from 1.6 to 18.8 of the atmospheric concentration, reflecting the enhanced contribution from baseflow and interflow influx as well as in situ oxidation of organic matter. However, in April and December, undersaturated pCO2 was found in some shallow, clean rivers in the upstream regions. delta13C of DIC has a narrow range from -9.07 to -13.59 per thousand, which was more depleted in the regions with metamorphic rocks and granites than in the carbonate regions. Seasonally, it was slightly more depleted in the dry season (December) than in the wet season (June). The results suggested that lithological variability had a dominant control on spatial variations of water chemistry and carbon geochemistry in river systems. Besides, anthropogenic activities, such as agricultural and urban activities and in-stream damming, as well as river physical properties, such as water depth and transparency, also indicated their impacts. The seasonal variations likely reflected the changes of hydrological regime, as well as metabolic processes in the river.

Geochemical characteristics and fluxes of organic carbon in a human-disturbed mountainous river (the Luodingjiang River) of the Zhujiang (Pearl River), China.

This study aims to investigate the state of the riverine organic carbon in the Luodingjiang River under human impacts, such as reforestation, construction of reservoirs and in-stream damming. Seasonal and spatial characteristics of total suspended sediment (TSS), dissolved organic carbon (DOC) and particulate organic carbon (POC), as well as C/N ratios and the stable carbon isotopic signatures of POC (delta(13)C(POC)) were examined based on a one-year study (2005) in the basin-wide scale. More frequent sampling was conducted in the outlet of the river basin at Guanliang hydrological station. DOC and POC concentrations showed flush effects with increasing water discharge and sediment load in the basin-wide scale. Atomic C/N ratio of POC had a positive relationship with TSS in the outlet of the basin, indicating the reduced aquatic sources and enhanced terrestrial sources during the high flood season. However, the similar relationship was not observed in the basin-wide scale mainly due to the spatial distributions of soil organic carbon and TSS. delta(13)C(POC) showed obvious seasonal variations with enriched values in the period with high TSS concentration, reflecting the increased contribution from C(4) plants with enhanced soil erosion. The specific flux of the total organic carbon (2.30 t km(-)(2) year(-1)) was smaller than the global average level. The ratio of DOC to POC was 1.17, which is higher than most rivers under Asian monsoon climate regime. The organic carbon flux was estimated to decline with decreasing sediment load as a result of reforestation, reservoir construction and in-stream damming, which demonstrates the impacts of human disturbances on the global carbon cycle.

Depth-sensitive Raman spectroscopy for skin wound evaluation in rodents.

Raman spectroscopy has demonstrated great potential for skin wound assessment. Given that biochemical changes in wound healing is depth dependent as the skin is a layered structure, depth sensitive Raman spectroscopy could enhance the power of Raman spectroscopy in this application. Considering the critical importance of rodent studies in the field of skin wound assessment, it is necessary to develop and validate a system that can perform depth sensitive measurements in rat skin with a proper target depth range. In this manuscript, we report the design, optimization and evaluation of a new snapshot depth-sensitive Raman instrument for rat skin measurements. The optical design and optimization process are presented first. The depth sensitive measurement performance is characterized on both ex vivo porcine skin with a gradient of layer thickness and ex vivo rat skin samples with wounds. The statistical analysis of the measured Raman spectra demonstrates the feasibility of differentiation between the wound edge and healthy skin. Moreover, the accuracy of classification improves monotonically as more data from new depths are used, which implies that each depth offers additional information useful for classification. This instrument demonstrates the ability to perform snapshot depth sensitive Raman measurements from rat skin, which paves the way towards in vivo preclinical studies of rat skin wounds.

The Potential Impact of Connexin 43 Expression on Bcl-2 Protein Level and Taxane Sensitivity in Head and Neck Cancers-In Vitro Studies.

The poor prognosis of head and neck squamous cell carcinoma (HNSCC) is partly due to the lack of reliable predictive markers. Connexin 43 (Cx43) protein and its cell-communication channels have been assigned tumor suppressor functions while the anti-apoptotic Bcl-2 (B-cell lymphoma-2) protein has been associated with negative prognostic significance in cancer. This study aimed to test the role of Cx43 protein on Bcl-2 expression, tumor progression and response to taxane-based treatment in HNSCC. Human papillomavirus (HPV) negative HNSCC cell lines were tested for paclitaxel sensitivity through measuring apoptosis induction, cell viability and changes in Cx43 and Bcl-2 levels using flow cytometry, cell viability assay, immunocytochemistry and western blot. Inhibition of Cx43 expression using siRNA increased Bcl-2 protein levels in SCC25 (tongue squamous cell carcinoma) cells, while forced Cx43 expression reduced Bcl-2 levels and supported paclitaxel cytotoxicity in FaDu (hypopharynx squamous cell carcinoma) cells. In vitro results were in line with protein expression and clinicopathological features tested in tissue microarray samples of HNSCC patients. Our data demonstrate that elevated Cx43 and reduced Bcl-2 levels may indicate HNSCC sensitivity to taxane-based treatments. On the contrary, silencing of the Cx43 gene GJA1 (gap junction protein alpha-1) can result in increased Bcl-2 expression and reduced paclitaxel efficiency. Clinical tumor-based analysis also confirmed the inverse correlation between Cx43 and Bcl-2 expression.

Molecular mechanisms of subtype-specific inhibition of neuronal T-type calcium channels by ascorbate.

T-type Ca2+ channels (T-channels) are involved in the control of neuronal excitability and their gating can be modulated by a variety of redox agents. Ascorbate is an endogenous redox agent that can function as both an anti- and pro-oxidant. Here, we show that ascorbate selectively inhibits native Ca(v)3.2 T-channels in peripheral and central neurons, as well as recombinant Ca(v)3.2 channels heterologously expressed in human embryonic kidney 293 cells, by initiating the metal-catalyzed oxidation of a specific, metal-binding histidine residue in domain 1 of the channel. Our biophysical experiments indicate that ascorbate reduces the availability of Ca(v)3.2 channels over a wide range of membrane potentials, and inhibits Ca(v)3.2-dependent low-threshold-Ca2+ spikes as well as burst-firing in reticular thalamic neurons at physiologically relevant concentrations. This study represents the first mechanistic demonstration of ion channel modulation by ascorbate, and suggests that ascorbate may function as an endogenous modulator of neuronal excitability.

Australian Lamb Meat - The Response to Societal and Ethnic Influences.

Lamb has long been considered a traditional meal within Australia; however as consumer preferences have changed since the 1950's, consumption of lamb has decreased from the 1980's. This is the result of changing societal roles, particularly for females, decreasing household sizes and increasing awareness of the impact of food choices on human health. Since the 1980's improvement of farm practices and increases in genetic gains has addressed part of this decline by increasing the amount of lean meat and decreasing fat in lamb retail cuts. Yet, this has created a challenge for the industry to utilise the larger carcases now being produced. Thus, a whole value chain approach to increasing consumption has been undertaken through several research programs to create cuts which suit the modern consumer, examine nutritional and eating quality and increase adoption of value added cuts. Therefore, this paper outlines this history of changing consumer patterns and the consequent research to address these changes.

Connexins in endothelial barrier function - novel therapeutic targets countering vascular hyperpermeability.

Prolonged vascular hyperpermeability is a common feature of many diseases. Vascular hyperpermeability is typically associated with changes in the expression patterns of adherens and tight junction proteins. Here, we focus on the less-appreciated contribution of gap junction proteins (connexins) to basal vascular permeability and endothelial dysfunction. First, we assess the association of connexins with endothelial barrier integrity by introducing tools used in connexin biology and relating the findings to customary readouts in vascular biology. Second, we explore potential mechanistic ties between connexins and junction regulation. Third, we review the role of connexins in microvascular organisation and development, focusing on interactions of the endothelium with mural cells and tissue-specific perivascular cells. Last, we see how connexins contribute to the interactions between the endothelium and components of the immune system, by using neutrophils as an example. Mounting evidence of crosstalk between connexins and other junction proteins suggests that we rethink the way in which different junction components contribute to endothelial barrier function. Given the multiple points of connexin-mediated communication arising from the endothelium, there is great potential for synergism between connexin-targeted inhibitors and existing immune-targeted therapeutics. As more drugs targeting connexins progress through clinical trials, it is hoped that some might prove effective at countering vascular hyperpermeability.

An Algorithm for Neuropathic Pain Management in Older People.

Neuropathic pain frequently affects older people, who generally also have several comorbidities. Elderly patients are often poly-medicated, which increases the risk of drug-drug interactions. These patients, especially those with cognitive problems, may also have restricted communication skills, making pain evaluation difficult and pain treatment challenging. Clinicians and other healthcare providers need a decisional algorithm to optimize the recognition and management of neuropathic pain. We present a decisional algorithm developed by a multidisciplinary group of experts, which focuses on pain assessment and therapeutic options for the management of neuropathic pain, particularly in the elderly. The algorithm involves four main steps: (1) detection, (2) evaluation, (3) treatment, and (4) re-evaluation. The detection of neuropathic pain is an essential step in ensuring successful management. The extent of the impact of the neuropathic pain is then assessed, generally with self-report scales, except in patients with communication difficulties who can be assessed using behavioral scales. The management of neuropathic pain frequently requires combination treatments, and recommended treatments should be prescribed with caution in these elderly patients, taking into consideration their comorbidities and potential drug-drug interactions and adverse events. This algorithm can be used in the management of neuropathic pain in the elderly to ensure timely and adequate treatment by a multidisciplinary team.

Postictal behavioural impairments are due to a severe prolonged hypoperfusion/hypoxia event that is COX-2 dependent.

Seizures are often followed by sensory, cognitive or motor impairments during the postictal phase that show striking similarity to transient hypoxic/ischemic attacks. Here we show that seizures result in a severe hypoxic attack confined to the postictal period. We measured brain oxygenation in localized areas from freely-moving rodents and discovered a severe hypoxic event (pO2 < 10 mmHg) after the termination of seizures. This event lasted over an hour, is mediated by hypoperfusion, generalizes to people with epilepsy, and is attenuated by inhibiting cyclooxygenase-2 or L-type calcium channels. Using inhibitors of these targets we separated the seizure from the resulting severe hypoxia and show that structure specific postictal memory and behavioral impairments are the consequence of this severe hypoperfusion/hypoxic event. Thus, epilepsy is much more than a disease hallmarked by seizures, since the occurrence of postictal hypoperfusion/hypoxia results in a separate set of neurological consequences that are currently not being treated and are preventable.