Preconception opioids interact with mouse strain to alter morphine withdrawal in the next generation.

RATIONALE: Transgenerational effects of preconception morphine exposure in female rats have been reported which suggest that epigenetic modifications triggered by female opioid exposure, even when that exposure ends several weeks prior to pregnancy, has significant ramifications for their future offspring. OBJECTIVE: The current study compares two mouse strains with well-established genetic variation in their response to mu opioid receptor agonists, C57BL/6J (BL6) and 129S1/svlmJ (129) to determine whether genetic background modifies the impact of preconception opioid exposure. METHODS: Adolescent females from both strains were injected daily with morphine for a total of 10 days using an increasing dosing regimen with controls receiving saline. Several weeks after their final injection, aged-matched BL6 and 129 morphine (Mor-F0) or saline (Sal-F0) females were mated with drug naïve males to generate Mor-F1 and Sal-F1 offspring, respectively. As adults, F1 mice were made morphine dependent using thrice daily morphine injections for 4 days. On day 5, mice were administered either saline or morphine followed 3 h later by naloxone. Behavioral and physiological signs of withdrawal were then measured. RESULTS: Regardless of strain or sex, morphine-dependent Mor-F1 mice had significantly lower levels of withdrawal-induced corticosterone but significantly higher glucose levels when compared to Sal-F1 controls. In contrast, both strain- and preconception opioid exposure effects on physical signs of morphine dependence were observed.

Increased rates of suicide ideation and attempts in rural dwellers following the SARS-CoV-2 pandemic.

PURPOSE: Those factors identified to increase the risk of suicide in rural dwellers were exacerbated by the SARS-CoV-2 pandemic, specifically economic factors, substance use, access to health care, and access to lethal weapons. Because the effects of SARS-CoV-2 on suicide ideation and attempts in rural populations have not been fully characterized in published literature, this study compares: (1) the rates of suicide ideation and attempts between the 6 months affected by SARS-CoV-2 to same months of the preceding year (3/18/2020-9/18/20; 3/18/2019-9/18/19), (2) demographics (ie, age, sex, residence, race, and ethnicity), and (3) the locations in which the encounters were billed (inpatient, outpatient, and emergency department). METHODS: Deidentified claims data associated with patient encounters billed for Suicide Ideation and Suicide Attempt were grouped based on time period and analyzed using descriptive statistics, incidence rate ratio (IRR), 2-sample t-test, chi-square test of association, or Fisher's exact test. FINDINGS: Suicidal ideation encounters increased in the 6 months post-SARS-CoV-2 when compared to the 6 months of the prior year (IRR = 1.19; P < .001). Males (IRR = 1.27, P < .001), those residing rural areas (IRR = 1.22, P = .01), and Black, non-Hispanic (IRR = 1.24, P = .024) were found to have increased rates of suicide ideation post-SARS-Cov-2. In adults, White, non-Hispanics (IRR = 1.16; P < .001) had increased rates of post-SARS-CoV-2. In the pediatric subset, those who were aged 14-17 (IRR = 1.50; P < .001), resided in rural areas (IRR = 1.61, P = .009), and idenitifed as Hispanic (IRR = 1.89; P = .037) or Black, non-Hispanic (IRR = 1.61, P = .009) had increased rates post-SARS-CoV-2. CONCLUSIONS: Our study identified rural dwellers to be at increased risk for suicide ideation.

Recommendation for changes to the guidelines of trauma patients with potential spinal injury within a regional UK ambulance trust.

Background: Spinal assessment and immobilisation has been a topic of debate for many years where, despite an emerging evidence base and the delivery of new guidance overseas, little has changed within UK pre-hospital practice. Since 2018, South East Coast Ambulance Service NHS Foundation Trust has spent time working with local trauma networks and expertise from within the region and international colleagues to develop a set of C-spine assessment and immobilisation guidelines that reflect the current best available international evidence and significant changes in international pre-hospital practice from settings such as Scandinavia and Australasia. Methods: A specialist group was commissioned to review the topic of pre-hospital spinal immobilisation and explore potential for evidence-based improvement. In conjunction with local trauma networks, subject matter experts and a thorough review of recent literature, a series of recommendations were made in order to improve spinal care within the authoring trust. Results: Seven recommendations were made, and an updated set of guidelines produced. These included the removal of semi-rigid collars from pre-hospital spinal immobilisation; the creation of two tiers of patients to ensure that the high-risk and low-risk populations are considered separately and an accompanying decision tool to safeguard both cohorts; an increased emphasis on the risk of spinal injury in the frail and older patient; an emphasis on spinal motion restriction rather than rigid immobilisation; an increased emphasis on self-extrication; and the use of a marker for emergency departments. Summary: An updated set of guidance has been produced using a combination of specialist and expert opinion alongside a literature review with close involvement of key stakeholders, both public and professional. The new guidance helps to ensure a patient-centred approach where each person is considered an individual with their risk of injury and management measures tailored to their specific needs.

Astrocytes display cell autonomous and diverse early reactive states in familial amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis is a rapidly progressive and fatal disease. Although astrocytes are increasingly recognized contributors to the underlying pathogenesis, the cellular autonomy and uniformity of astrocyte reactive transformation in different genetic forms of amyotrophic lateral sclerosis remain unresolved. Here we systematically examine these issues by using highly enriched and human induced pluripotent stem cell-derived astrocytes from patients with VCP and SOD1 mutations. We show that VCP mutant astrocytes undergo cell-autonomous reactive transformation characterized by increased expression of complement component 3 (C3) in addition to several characteristic gene expression changes. We then demonstrate that isochronic SOD1 mutant astrocytes also undergo a cell-autonomous reactive transformation, but that this is molecularly distinct from VCP mutant astrocytes. This is shown through transcriptome-wide analyses, identifying divergent gene expression profiles and activation of different key transcription factors in SOD1 and VCP mutant human induced pluripotent stem cell-derived astrocytes. Finally, we show functional differences in the basal cytokine secretome between VCP and SOD1 mutant human induced pluripotent stem cell-derived astrocytes. Our data therefore reveal that reactive transformation can occur cell autonomously in human amyotrophic lateral sclerosis astrocytes and with a striking degree of early molecular and functional heterogeneity when comparing different disease-causing mutations. These insights may be important when considering astrocyte reactivity as a putative therapeutic target in familial amyotrophic lateral sclerosis.

Randomised methodology development study to investigate plaque removal efficacy of manual toothbrushes.

OBJECTIVES: This study aimed to evaluate plaque removal efficacy of toothbrushes to guide clinical model development. METHODS: This single-centre, randomised, controlled, examiner-blind, method development study included 80 healthy participants. Over 28 days, participants brushed twice-daily using a fluoride toothpaste and one of four marketed toothbrushes: Toothbrushes A (compact head) and B (regular head): medium-hard; flat trim; end-rounded bristles; Toothbrush C: medium-hard; end-rounded bristles; silky, tapered filaments; compact head; Toothbrush D: soft; tapered filaments; compact head. Supra-gingival plaque removal was evaluated immediately after single brushing events (Days 0, 7, 28, at study centre) and following 7- and 28-days' home use via Rustogi modified Navy Plaque Index (RPI) and Turesky modified Quigley Hein Plaque Index (TPI). RESULTS: All toothbrushes provided significant plaque removal after single-brushing events with change from Day 0 pre-brushing scores on RPI ranging from -0.10 to -0.16 (p<.0001) and on TPI ranging from -0.61 to -0.89 (p<.0001). Toothbrushes A and B showed significant (p<.05) pre-brushing RPI/TPI plaque reductions after 7- (-0.04/-0.06, respectively for RPI; -0.16/-0.20, respectively for TPI) and 28-days (-0.04/-0.03, respectively for RPI; -0.20/-0.11, respectively for TPI) use versus Day 0 pre-brushing (except Toothbrush B, Day 28, non-significant TPI). There were no significant differences with Toothbrush C. Toothbrush D TPI was significantly lower at both timepoints versus Day 0 pre-brushing (p<.05). Study toothbrushes were generally well-tolerated. CONCLUSION: The observations from this study showed how various aspects of a study design could impact toothbrushes performance. These data will inform the design of future clinical studies of plaque removal efficacy using manual toothbrushes. CLINICAL SIGNIFICANCE STATEMENT: Regular effective oral hygiene can help prevent and treat gingivitis, principally via twice-daily mechanical cleaning with a toothbrush. Data generated from this methodology development study will help to identify the key aspects which impact toothbrushes' performance and understand which one would be more suitable to answer questions of scientific interest. This study provides useful information for the design of future clinical trials to assess plaque removal efficacy of manual toothbrushes and generate results to inform clinical recommendations.

The UPTAKE study: a cross-sectional survey examining the insights and beliefs of the UK population on COVID-19 vaccine uptake and hesitancy.

OBJECTIVE: A key challenge towards a successful COVID-19 vaccine uptake is vaccine hesitancy. We examine and provide novel insights on the key drivers and barriers towards COVID-19 vaccine uptake. DESIGN: This study involved an anonymous cross-sectional online survey circulated across the UK in September 2020. The survey was designed to include several sections to collect demographic data and responses on (1) extent of agreement regarding various statements about COVID-19 and vaccinations, (2) previous vaccination habits (eg, if they had previously declined vaccination) and (3) interest in participation in vaccine trials. Multinominal logistic models examined demographic factors that may impact vaccine uptake. We used principle component analysis and text mining to explore perception related to vaccine uptake. SETTING: The survey was circulated through various media, including posts on social media networks (Facebook, Twitter, LinkedIn and Instagram), national radio, news articles, Clinical Research Network website and newsletter, and through 150 West Midlands general practices via a text messaging service. PARTICIPANTS: There were a total of 4884 respondents of which 9.44% were black, Asian and minority ethnic (BAME) group. The majority were women (n=3416, 69.9%) and of white ethnicity (n=4127, 84.5%). RESULTS: Regarding respondents, overall, 3873 (79.3%) were interested in taking approved COVID-19 vaccines, while 677 (13.9%) were unsure, and 334 (6.8%) would not take a vaccine. Participants aged over 70 years old (OR=4.63) and the BAME community (OR=5.48) were more likely to take an approved vaccine. Smokers (OR=0.45) and respondents with no known illness (OR=0.70) were less likely to accept approved vaccines. The study identified 16 key reasons for not accepting approved vaccines, the most common (60%) being the possibility of the COVID-19 vaccine having side effects. CONCLUSIONS: This study provides an insight into focusing on specific populations to reduce vaccine hesitancy. This proves crucial in managing the COVID-19 pandemic.

The UPTAKE study: implications for the future of COVID-19 vaccination trial recruitment in UK and beyond.

BACKGROUND: Developing a safe and effective vaccine will be the principal way of controlling the COVID-19 pandemic. However, current COVID-19 vaccination trials are not adequately representing a diverse participant population in terms of age, ethnicity and comorbidities. Achieving the representative recruitment targets that are adequately powered to the study remains one of the greatest challenges in clinical trial management. To ensure accuracy and generalisability of the safety and efficacy conclusions generated by clinical trials, it is crucial to recruit patient cohorts as representative as possible of the future target population. Missing these targets can lead to reduced validity of the study results and can often slow down drug development leading to costly delays. OBJECTIVE: This study explores the key factors related to perceptions and participation in vaccination trials. METHODS: This study involved an anonymous cross-sectional online survey circulated across the UK. Statistical analysis was done in six phases. Multi-nominal logistic models examined demographic and geographic factors that may impact vaccine uptake. RESULTS: The survey had 4884 participants of which 9.44% were Black Asian Minority Ethnic (BAME). Overall, 2020 (41.4%) respondents were interested in participating in vaccine trials; 27.6% of the respondents were not interested and 31.1% were unsure. The most interested groups were male (OR = 1.29), graduates (OR = 1.28), the 40-49 and 50-59 age groups (OR = 1.88 and OR = 1.46 respectively) and those with no health issues (OR = 1.06). The least interested groups were BAME (OR = 0.43), those from villages and small towns (OR = 0.66 and 0.54 respectively) and those aged 70 and above (OR = 1.11). CONCLUSIONS: In order to have a vaccination that is generalisable to the entire population, greater work needs to be done in engaging a diverse cohort of participants. Public health campaigns need to be targeted in improving trial recruitment rates for the elderly, BAME community and the less educated rural population.

Hub-and-spoke model for thrombectomy service in UK NHS practice.

Mechanical thrombectomy is a highly effective but time dependent treatment for acute ischaemic stroke due to large vessel occlusion. In the UK, the national clinical guidelines for stroke and National Institute for Health and Care Excellence guidance endorses thrombectomy as an acute stroke treatment, and NHS England commissioned thrombectomy services. However, there are no UK 'real-world' data to verify the efficacy of the hub-and-spoke model in thrombectomy. There are currently 24 tertiary neuroscience centres in the UK that can provide thrombectomy treatment and many of these operate only within working hours. This study is the first to demonstrate that a hub-and-spoke thrombectomy service in routine UK 24/7 clinical practice is as effective and safe as in the setting of randomised controlled clinical trials. However, there are 9.3% of patients accepted for transfer to the thrombectomy centre who did not proceed to thrombectomy, mostly due to delays. Fifty-three per cent of thrombectomy cases were performed outside of standard working hours when transfer delays were increased. A 24/7 thrombectomy service is needed to maximise the benefit to all suitable patients. Measures, including improving workflow and optimising work forces, are needed to minimise the delays and continue to improve the service.

Delineating Astrocytic Cytokine Responses in a Human Stem Cell Model of Neural Trauma.

Neuroinflammation has been shown to mediate the pathophysiological response following traumatic brain injury (TBI). Accumulating evidence implicates astrocytes as key immune cells within the central nervous system (CNS), displaying both pro- and anti-inflammatory properties. The aim of this study was to investigate how in vitro human astrocyte cultures respond to cytokines across a concentration range that approximates the aftermath of human TBI. To this end, enriched cultures of human induced pluripotent stem cell (iPSC)-derived astrocytes were exposed to interleukin-1ß (IL-1ß) (1-10,000 pg/mL), IL-4 (1-10,000 pg/mL), IL-6 (100-1,000,000 pg/mL), IL-10 (1-10,000 pg/mL) and tumor necrosis factor (TNF)-α (1-10,000 pg/mL). After 1, 24, 48 and 72 h, cultures were fixed and immunolabeled, and the secretome/supernatant was analyzed at 24, 48, and 72 h using a human cytokine/chemokine 39-plex Luminex assay. Data were compared to previous in vitro studies of neuronal cultures and clinical TBI studies. The secretome revealed concentration-, time- and/or both concentration- and time-dependent production of downstream cytokines (29, 21, and 17 cytokines, respectively, p<0.05). IL-1ß exposure generated the most profound downstream response (27 cytokines), IL-6 and TNF had intermediate responses (13 and 11 cytokines, respectively), whereas IL-4 and IL-10 only led to weak responses over time or in escalating concentration (8 and 8 cytokines, respectively). Notably, expression of IL-1ß, IL-6, and TNF cytokine receptor mRNA was higher in astrocyte cultures than in neuronal cultures. Several secreted cytokines had temporal trajectories, which corresponded to those seen in the aftermath of human TBI. In summary, iPSC-derived astrocyte cultures exposed to cytokine concentrations reflecting those in TBI generated an increased downstream cytokine production, particularly IL-1ß. Although more work is needed to better understand how different cells in the CNS respond to the neuroinflammatory milieu after TBI, our data shows that iPSC-derived astrocytes represent a tractable model to study cytokine stimulation in a cell type-specific manner.

Low Incidence of Pelvic Sepsis after Hartmann's Procedure: Radiation Therapy May Be a Risk Factor.

PURPOSE: Hartmann's procedure is a well-established alternative in colorectal surgery when a primary anastomosis is contraindicated. However, the rectal remnant may cause complications. This study was designed to investigate the occurrence of pelvic sepsis after Hartmann's procedure and identify possible risk factors. METHODS: All patients who underwent Hartmann's procedure between 2005 and 2012 were identified by the in-hospital registry. Information about pelvic sepsis and potential preoperative, perioperative, and postoperative risk factors was obtained by review of the medical records. RESULTS: 172 patients were identified (97 females); they were aged 74 ± 11 years. Surgery was performed due to cancer (49%) or diverticulitis (35%) and other benign disease (16%). Rectal transection was carried out anywhere between the pelvic floor and the promontory. Pelvic sepsis developed in 6.4% (11/172) of patients. Pelvic sepsis was associated with preoperative radiotherapy (p = 0.03) and Hinchey grade III and IV (p = 0.02) in those patients who underwent Hartmann's procedure for diverticular disease. CONCLUSION: Hartmann's procedure is a safe operation when an anastomosis is contraindicated since the incidence of pelvic sepsis is low. Preoperative radiotherapy and Hinchey grade III and IV may be risk factors for the development of pelvic sepsis.

Efficacy of a 3% potassium nitrate mouthrinse for the relief of dentinal hypersensitivity: An 8-week randomized controlled study.

BACKGROUND: Mouthrinses containing potassium salts have been shown to be effective for the relief of dentinal hypersensitivity (DH) when used adjunctively to toothbrushing with a nonsensitivity toothpaste. METHODS: The authors conducted a randomized, 8-week, single-center, examiner-blinded, parallel-group clinical trial with 191 participants with DH. Participants were randomized to twice-daily use of either 3% potassium nitrate (KNO3) mouthrinse plus fluoride toothpaste or the same fluoride toothpaste alone. The primary outcome was change from baseline in response to an evaporative (air) stimulus at 8 weeks, measured using the Schiff sensitivity scale. Secondary outcomes were response to an evaporative (air) stimulus with the Schiff sensitivity scale (4 weeks) and a visual rating scale (4 and 8 weeks) and response to a tactile stimulus (4 and 8 weeks). RESULTS: Both groups showed statistically significant improvements in evaporative (air) sensitivity from baseline after 4 and 8 weeks (P < .0001). At weeks 4 and 8, the authors observed significant improvements from baseline in tactile sensitivity only in the KNO3 mouthrinse group (P < .0001). Between-treatment comparisons for all sensitivity measures at both time points showed statistically significantly greater DH reductions in the KNO3 mouthrinse group compared with the toothpaste-alone group (P = .0004 for the visual rating scale at week 4; P < .0001 for all other measures and time points). Treatments were generally well tolerated. CONCLUSIONS: Twice-daily use of a 3% KNO3 mouthrinse, adjunctive to toothbrushing with fluoride toothpaste, provided significant improvements in DH compared with fluoride toothpaste alone. PRACTICAL IMPLICATIONS: Addition of 3% KNO3 mouthrinse to a typical oral hygiene regimen of toothbrushing with fluoride toothpaste provides an alternative strategy for the management of DH. ClinicalTrials.gov: NCT02226562.

What is the Impact of Volunteers Providing Care and Support for People with Dementia in Acute Hospitals? A Systematic Review.

A quarter of acute hospital beds are occupied by people with dementia, and a hospital stay may impact negatively on their health and wellbeing. The development and implementation of volunteers to provide social, activity-based, one-to-one support for people with dementia in acute hospitals has become routine practice. However, the evidence to support this practice has not been identified or evaluated. This systematic review considers the effect of volunteers on the care and experience of people with co-morbid cognitive impairment/dementia in acute hospitals. The systematic search identified 444 papers, although only three papers included specific analysis relating to the impact of volunteers. The evidence suggests volunteers may have potential to enhance the experiences of people with dementia in acute hospitals; however, there is currently a marked lack of evidence to support the widespread implementation of volunteers. There is therefore an urgent need for multi-site robust research to provide evidence of the impact of volunteers supporting people with cognitive impairment/dementia during an acute hospital stay.

A Review of the Role of Carcinoembryonic Antigen in Clinical Practice.

Carcinoembryonic antigen (CEA) is not normally produced in significant quantities after birth but is elevated in colorectal cancer. The aim of this review was to define the current role of CEA and how best to investigate patients with elevated CEA levels. A systematic review of CEA was performed, in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Studies were identified from PubMed, Cochrane library, and controlled trials registers. We identified 2,712 papers of which 34 were relevant. Analysis of these papers found higher preoperative CEA levels were associated with advanced or metastatic disease and thus poorer prognosis. Postoperatively, failure of CEA to return to normal was found to be indicative of residual or recurrent disease. However, measurement of CEA levels alone was not sufficient to improve survival rates. Two algorithms are proposed to guide investigation of patients with elevated CEA: one for patients with elevated CEA after CRC resection, and another for patients with de novo elevated CEA. CEA measurement has an important role in the investigation, management and follow-up of patients with colorectal cancer.

Intron retention and nuclear loss of SFPQ are molecular hallmarks of ALS.

Mutations causing amyotrophic lateral sclerosis (ALS) strongly implicate ubiquitously expressed regulators of RNA processing. To understand the molecular impact of ALS-causing mutations on neuronal development and disease, we analysed transcriptomes during in vitro differentiation of motor neurons (MNs) from human control and patient-specific VCP mutant induced-pluripotent stem cells (iPSCs). We identify increased intron retention (IR) as a dominant feature of the splicing programme during early neural differentiation. Importantly, IR occurs prematurely in VCP mutant cultures compared with control counterparts. These aberrant IR events are also seen in independent RNAseq data sets from SOD1- and FUS-mutant MNs. The most significant IR is seen in the SFPQ transcript. The SFPQ protein binds extensively to its retained intron, exhibits lower nuclear abundance in VCP mutant cultures and is lost from nuclei of MNs in mouse models and human sporadic ALS. Collectively, we demonstrate SFPQ IR and nuclear loss as molecular hallmarks of familial and sporadic ALS.

Proteomic analysis defines kinase taxonomies specific for subtypes of breast cancer.

Multiplexed small molecule inhibitors covalently bound to Sepharose beads (MIBs) were used to capture functional kinases in luminal, HER2-enriched and triple negative (basal-like and claudin-low) breast cancer cell lines and tumors. Kinase MIB-binding profiles at baseline without perturbation proteomically distinguished the four breast cancer subtypes. Understudied kinases, whose disease associations and pharmacology are generally unexplored, were highly represented in MIB-binding taxonomies and are integrated into signaling subnetworks with kinases that have been previously well characterized in breast cancer. Computationally it was possible to define subtypes using profiles of less than 50 of the more than 300 kinases bound to MIBs that included understudied as well as metabolic and lipid kinases. Furthermore, analysis of MIB-binding profiles established potential functional annotations for these understudied kinases. Thus, comprehensive MIBs-based capture of kinases provides a unique proteomics-based method for integration of poorly characterized kinases of the understudied kinome into functional subnetworks in breast cancer cells and tumors that is not possible using genomic strategies. The MIB-binding profiles readily defined subtype-selective differential adaptive kinome reprogramming in response to targeted kinase inhibition, demonstrating how MIB profiles can be used in determining dynamic kinome changes that result in subtype selective phenotypic state changes.

Galectin-1 Influences Breast Cancer Cell Adhesion to E-selectin Via Ligand Intermediaries.

INTRODUCTION: Invasion of other tissues during bloodborne metastasis in part requires adhesion of cancer cells to vascular endothelium by specific fluid shear-dependent receptor-ligand interactions. This study investigates the hypothesis that the adhesion is mediated by ligands shared between endothelial E-selectin and Galectin-1 (Gal-1), both of which are upregulated during inflammation and cancer. METHODS: Flow chamber adhesion and dynamic biochemical tissue analysis (DBTA) assays were used to evaluate whether Gal-1 modulates E-selectin adhesive interactions of breast cancer cells and tissues under dynamic flow conditions, while immunocytochemistry, immunohistochemistry, western blotting, and fluorescence anisotropy were used to study molecular interactions under static conditions. RESULTS: Dynamic adhesion assays revealed a shear-dependent binding interaction between Gal-1hFc treated breast cancer cells and tissues and E-selectin-coated beads, causing ~ 300% binding increase of the beads compared to negative controls. Immunocyto- and immunohistochemical analyses showed that Gal-1 and E-selectin fluorescent signals colocalized on cells and tissues at ~ 75% for each assay. Immunoprecipitation and Western blotting of Mac-2BP from breast cancer cell lysates revealed that Gal-1 and E-selectin share Mac-2BP as a ligand, while fluorescence anisotropy and circulating tumor cell model systems exhibited competitive or antagonistic binding between Gal-1 and E-selectin for shared ligands, including Mac-2BP. Furthermore, Mac-2BP functional blockade inhibited the effects of Gal-1 on E-selectin binding. CONCLUSIONS: In summary, this investigation reveals a shear-dependent interaction between E-selectin and Gal-1 that may be due to intermediation by a similar or shared ligand(s), including Mac-2BP, which may provide a rational basis for development of novel diagnostics or therapeutics for breast cancer.

Elucidating Pro-Inflammatory Cytokine Responses after Traumatic Brain Injury in a Human Stem Cell Model.

Cytokine mediated inflammation likely plays an important role in secondary pathology after traumatic brain injury (TBI). The aim of this study was to elucidate secondary cytokine responses in an in vitro enriched (>80%) human stem cell-derived neuronal model. We exposed neuronal cultures to pre-determined and clinically relevant pathophysiological levels of tumor necrosis factor-α (TNF), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß), shown to be present in the inflammatory aftermath of TBI. Data from this reductionist human model were then compared with our in vivo data. Human embryonic stem cell (hESC)-derived neurons were exposed to recombinant TNF (1-10,000 pg/mL), IL-1ß (1-10,000 pg/mL), and IL-6 (0.1-1000 ng/mL). After 1, 24, and 72 h, culture supernatant was sampled and analyzed using a human cytokine/chemokine 42-plex Milliplex kit on the Luminex platform. The culture secretome revealed both a dose- and/or time-dependent release of cytokines. The IL-6 and TNF exposure each resulted in significantly increased levels of >10 cytokines over time, while IL-1ß increased the level of C-X-C motif chemokine 10 (CXCL10/IP10) alone. Importantly, these patterns are consistent with our in vivo (human) TBI data, thus validating our human stem cell-derived neuronal platform as a clinically useful reductionist model. Our data cumulatively suggest that IL-6 and TNF have direct actions, while the action of IL-1ß on human neurons likely occurs indirectly through inflammatory cells. The hESC-derived neurons provide a valuable platform to model cytokine mediated inflammation and can provide important insights into the mechanisms of neuroinflammation after TBI.

Improved survival for rectal cancer compared to colon cancer: the four cohort study.

BACKGROUND: Colorectal cancer (CRC) is the third most common cancer worldwide. This study was undertaken to evaluate survival outcomes and changes of disease outcomes of CRC patients over the last decades. METHODS: A retrospective analysis of CRC patients in Christchurch was performed in four patient cohorts at 5 yearly intervals; 1993-94, 1998-99, 2004-05 and 2009. Data on cancer location, stage, surgical and oncological treatment and survival were collected. Univariate, multivariate and Kaplan-Meier survival analysis were performed. RESULTS: There were 1391 patients (355, 317, 419 and 300 per cohort), 1037 colon and 354 rectal cancers, respectively. For colon cancer, right-sided cancers appeared more common in later cohorts (P = 0.01). There was a significant decrease in the number of permanent stomas for colon cancer patients (P = 0.001). There was an analogous trend for rectal cancers (P = 0.075). More CRC patients with stage IV disease were treated surgically (P = 0.001) and colon cancer stages I and II tended to have increased survival if operated by a colorectal surgeon (P = 0.06). Oncology referrals have increased remarkably (P = 0.001). Overall 56% of patients were alive at 5 years however rectal cancer patients had significantly better 5-year survival than those with colon cancer (P < 0.05). DISCUSSION: This four cohort study shows that modern CRC survival continues to improve and is comparable to international standards. Furthermore, rectal cancer patients have a better 5-year survival than colon cancer patients. The improved survival with early stage colon cancers operated on by specialist colorectal surgeons needs further exploration.

Efficacy of an experimental 3% potassium nitrate mouthwash in providing long-term relief from dentin hypersensitivity: An 8-week randomized controlled study (Study 2).

PURPOSE: To evaluate the efficacy of an experimental mouthwash containing 3% potassium nitrate (KNO3) in the relief of dentin hypersensitivity when used as an adjunct to brushing with fluoride toothpaste compared with the use of the same toothpaste alone. METHODS: This was a randomized, two-treatment, examiner-blind, parallel-design single-center, 8-week study in healthy subjects with self-reported and clinically diagnosed dentin hypersensitivity. Subjects were randomized to receive either fluoride toothpaste plus 3.0% KNO2 mouthwash or the same fluoride toothpaste alone, and instructed to use their allocated treatment twice daily for the next 8 weeks. Dentin hypersensitivity was evaluated at baseline and following 4 and 8 weeks of treatment through assessment of responses to evaporative (air) and tactile stimuli [measured by the Schiff sensitivity scale/a visual rating scale (VRS) and tactile threshold, respectively], and using the Dentin Hypersensitivity Experience Questionnaire (DHEQ, a validated quality-of-life instrument for dentin hyper-sensitivity). RESULTS: A total of 135 subjects were randomized and all completed the study. Both treatment groups demonstrated statistically significant improvements in sensitivity from baseline for each clinical measure of sensitivity (P< 0.0001) at Week 4 and Week 8. The toothpaste plus mouthwash group showed greater reductions in sensitivity at both timepoints for all clinical measures; between-treatment differences were only statistically significant for responses to an evaporative (air) stimulus (Schiff sensitivity score and VRS) at Week 4. There was evidence of an improvement in dentin hypersensitivity-associated quality of life as measured by changes from baseline in several DHEQ parameters for both treatment groups, but there were no statistically significant differences between treatments. CLINICAL SIGNIFICANCE: Although in the current study adjunctive use of a 3% KNO2 mouthwash did not provide statistically significant improvements in dentin hypersensitivity for all clinical measures at all timepoints compared with use of fluoride toothpaste alone, the reductions in sensitivity observed in this study are compatible with the findings of a previous study that showed adjunctive use of a 3% KNO2 mouthwash to be effective in providing relief from dentin hypersensitivity after 8 weeks' twice-daily use.

Efficacy of an experimental 3% potassium nitrate mouthwash in providing long-term relief from dentin hypersensitivity:An 8-week randomized controlled study (Study 1).

PURPOSE: To evaluate the efficacy of an experimental mouthwash containing 3% potassium nitrate (KNO3) in relieving dentin hypersensitivity when used as an adjunct to brushing with fluoride toothpaste compared with use of the same toothpaste alone. METHODS: This was one of three randomized, two-treatment, examiner-blind, parallel-design, single-site, 8-week studies in healthy subjects with self-reported and clinically diagnosed dentin hypersensitivity. Subjects were randomized to receive either fluoride toothpaste plus 3% KNO3 mouthwash or the same fluoride toothpaste alone, and instructed to use their allocated treatment twice daily for the next 8 weeks. Dentin hypersensitivity was evaluated at baseline and following 4 and 8 weeks of treatment through assessment of responses to evaporative (air) and tactile stimuli [measured by the Schiff Sensitivity Scale, a visual rating scale (VRS), and tactile threshold, respectively], and using the Dentin Hypersensitivity Experience Questionnaire (DHEQ; a validated quality-of-life instrument for dentin hypersensitivity). RESULTS: A total of 216 subjects were randomized and 214 completed the study. Both treatment groups demonstrated statistically significant reductions from baseline for each clinical measure of sensitivity (P≤ 0.01) at Weeks 4 and 8. Use of the 3% KNO3 mouthwash after brushing with fluoride toothpaste resulted in statistically significantly greater reductions in sensitivity to an evaporative (air) stimulus (mean Schiff score and mean VRS, P< 0.001; primary objective mean Schiff score at Week 8, P< 0.0001) and statistically significantly higher tactile threshold (P< 0.001) at Weeks 4 and 8 compared with toothpaste alone. The DHEQ responses reflected the clinical outcomes for several parameters, indicating a significant improvement in oral health-related quality of life after 8 weeks' use of the 3% KNO3 mouthwash. CLINICAL SIGNIFICANCE: The results of this study suggest that daily use of a 3% KNO3 mouthwash as an adjunct to brushing with fluoride toothpaste provides clinically relevant improvements in dentin hypersensitivity after 8 weeks' twice-daily use.