The Stickland Reaction Precursor trans-4-Hydroxy-l-Proline Differentially Impacts the Metabolism of Clostridioides difficile and Commensal Clostridia.

An intact gut microbiota confers colonization resistance against Clostridioides difficile through a variety of mechanisms, likely including competition for nutrients. Recently, proline was identified as an important environmental amino acid that C. difficile uses to support growth and cause significant disease. A posttranslationally modified form, trans-4-hydroxyproline, is highly abundant in collagen, which is degraded by host proteases in response to C. difficile toxin activity. The ability to dehydrate trans-4-hydroxyproline via the HypD glycyl radical enzyme is widespread among gut microbiota, including C. difficile and members of the commensal Clostridia, suggesting that this amino acid is an important nutrient in the host environment. Therefore, we constructed a C. difficile ΔhypD mutant and found that it was modestly impaired in fitness in a mouse model of infection, and was associated with an altered microbiota when compared to mice challenged with the wild-type strain. Changes in the microbiota between the two groups were largely driven by members of the Lachnospiraceae family and the Clostridium genus. We found that C. difficile and type strains of three commensal Clostridia had significant alterations to their metabolic gene expression in the presence of trans-4-hydroxyproline in vitro. The proline reductase (prd) genes were elevated in C. difficile, consistent with the hypothesis that trans-4-hydroxyproline is used by C. difficile to supply proline for energy metabolism. Similar transcripts were also elevated in some commensal Clostridia tested, although each strain responded differently. This suggests that the uptake and utilization of other nutrients by the commensal Clostridia may be affected by trans-4-hydroxyproline metabolism, highlighting how a common nutrient may be a signal to each organism to adapt to a unique niche. Further elucidation of the differences between them in the presence of hydroxyproline and other key nutrients will be important in determining their role in nutrient competition against C. difficile. IMPORTANCE Proline is an essential environmental amino acid that C. difficile uses to support growth and cause significant disease. A posttranslationally modified form, hydroxyproline, is highly abundant in collagen, which is degraded by host proteases in response to C. difficile toxin activity. The ability to dehydrate hydroxyproline via the HypD glycyl radical enzyme is widespread among gut microbiota, including C. difficile and members of the commensal Clostridia, suggesting that this amino acid is an important nutrient in the host environment. We found that C. difficile and three commensal Clostridia strains had significant, but different, alterations to their metabolic gene expression in the presence of hydroxyproline in vitro. This suggests that the uptake and utilization of other nutrients by the commensal Clostridia may be affected by hydroxyproline metabolism, highlighting how a common nutrient may be a signal to each organism to adapt to a unique niche. Further elucidation of the differences between them in the presence of hydroxyproline and other key nutrients will be important to determining their role in nutrient competition against C. difficile.

Parkinson's Disease Master Regulators on Substantia Nigra and Frontal Cortex and Their Use for Drug Repositioning.

Parkinson's disease (PD) is among the most prevalent neurodegenerative diseases. Available evidences support the view of PD as a complex disease, being the outcome of interactions between genetic and environmental factors. In face of diagnosis and therapy challenges, and the elusive PD etiology, the use of alternative methodological approaches for the elucidation of the disease pathophysiological mechanisms and proposal of novel potential therapeutic interventions has become increasingly necessary. In the present study, we first reconstructed the transcriptional regulatory networks (TN), centered on transcription factors (TF), of two brain regions affected in PD, the substantia nigra pars compacta (SNc) and the frontal cortex (FCtx). Then, we used case-control studies data from these regions to identify TFs working as master regulators (MR) of the disease, based on region-specific TNs. Twenty-nine regulatory units enriched with differentially expressed genes were identified for the SNc, and twenty for the FCtx, all of which were considered MR candidates for PD. Three consensus MR candidates were found for SNc and FCtx, namely ATF2, SLC30A9, and ZFP69B. In order to search for novel potential therapeutic interventions, we used these consensus MR candidate signatures as input to the Connectivity Map (CMap), a computational drug repositioning webtool. This analysis resulted in the identification of four drugs that reverse the expression pattern of all three MR consensus simultaneously, benperidol, harmaline, tubocurarine chloride, and vorinostat, thus suggested as novel potential PD therapeutic interventions.

The effect of DNA recovery on the subsequent quality of latent fingermarks: A pseudo-operational trial.

The recovery of fingermarks and DNA from the same location at a crime scene can be problematic because of contamination issues associated with powdering or laboratory-based visualisation processes and/or the perceived destructive impact of commonly employed 'swabbing' approaches to DNA recovery. Previous research in a controlled environment demonstrated that it was possible to recover DNA and latent fingermarks from the same location on various substrates when an adhesive approach to DNA recovery was used. The aim of this research was to conduct a pseudo-operational trial into the dual recovery of DNA and fingermarks using gel lifters for DNA recovery. Participants were asked to voluntarily and anonymously donate a wide variety of porous and non-porous substrates post handling. No instruction as to fingermark deposition nor environmental storage was provided. BVDA gel lifters were applied to the substrates to replicate DNA recovery followed by the application of fingermark visualisation processes. The number and quality of the fingermarks was established using a grading approach. Application factors were also investigated to consider the effects of user variation. The results demonstrated that it was possible to recover DNA and fingermarks considered to be capable of supporting an identification. Fingermark quality post lifting was dependant on the substrates used. The weight applied to the gel during its application was a lesser contributing factor than the duration of its contact with the surface. There was a greater chance of leaving the fingermarks unaltered with the application of a low weight and instantaneous retraction.

Undiscovered Pulsar in the Local Bubble as an Explanation of the Local High Energy Cosmic Ray All-Electron Spectrum.

Cosmic ray electrons and positrons are tracers of particle propagation in the interstellar medium (ISM). A recent measurement performed using the High Energy Stereoscopic System extends the all-electron (electron+positron) spectrum up to 20 TeV, probing very local sources and transport due to the ∼10 kyr cooling time of these particles. An additional key local measurement was the recent estimation of the ISM diffusion coefficient around Geminga performed using the High-Altitude Water Cherenkov Observatory. The inferred diffusion coefficient is much lower than typically assumed values. It has been argued that if this diffusion coefficient is representative of the local ISM, pulsars would not be able to account for the all-electron spectrum measured at Earth. Here we show that a low diffusion coefficient in the local ISM is compatible with a pulsar wind nebula origin of the highest energy electrons, if a so-far-undiscovered pulsar with spin-down power ∼10^{33-34} erg/s exists within 30-80 pc of Earth. The existence of such a pulsar is broadly consistent with the known population and may be detected in near future survey observations.

Successful pharmacotherapy for the treatment of severe feeding aversion with mechanistic insights from cross-species neuronal remodeling.

Pediatric feeding disorders affect up to 5% of children, causing severe food intake problems that can result in serious medical and developmental outcomes. Behavioral intervention (BI) is effective in extinguishing feeding aversions, and also expert-dependent, time/labor-intensive and not well understood at a neurobiological level. Here we first conducted a double-blind, placebo-controlled trial comparing BI with BI plus d-cycloserine (DCS). DCS is a partial N-methyl-d-aspartate (NMDA) receptor agonist shown to augment extinction therapies in multiple anxiety disorders. We examined whether DCS enhanced extinction of feeding aversion in 15 children with avoidant/restrictive food intake disorder (ages 20-58 months). After five treatment days, BI improved feeding by 37%. By contrast, BI+DCS improved feeding by 76%. To gain insight into possible mechanisms of successful intervention, we next tested the neurobiological consequences of DCS in a murine model of feeding aversion and avoidance. In mice with conditioned food aversion, DCS enhanced avoidance extinction across a broad dose range. Confocal fluorescence microscopy and three-dimensional neuronal reconstruction indicated that DCS enlarged dendritic spine heads-the primary sites of excitatory plasticity in the brain-within the orbitofrontal prefrontal cortex, a sensory-cognition integration hub. DCS also increased phosphorylation of the plasticity-associated extracellular signal-regulated kinase 1/2. In summary, DCS successfully augments the extinction of food aversion in children and mice, an effect that may involve plasticity in the orbitofrontal cortex. These results warrant a larger-scale efficacy study of DCS for the treatment of pediatric feeding disorders and further investigations of neural mechanisms.

Prevention of postsurgical wound dehiscence after abdominal surgery with NPWT: a multicentre randomised controlled trial protocol.

OBJECTIVE: The effectiveness of negative pressure wound therapy (NPWT) in the prevention of postoperative surgical wound dehiscence (SWD) is the subject of much debate and remains to be determined. This study will identify individuals at risk of postoperative SWD and trial the use of NPWT as a prophylactic measure against the occurrence of SWD, compared with a non-NPWT standard surgical dressing (SSD). METHOD: A prospective multicentre randomised controlled trial comparing NPWT dressing against standard surgical dressings (SSD) will be conducted. An intention-to-treat (ITT) approach will be used for the trial. AIMS: The primary outcome is the prevention of postoperative SWD up to and including day 30 postoperative. Secondary outcomes are: prevention of surgical site infection (SSI) and economic analysis of treatment groups. CONCLUSION: This study will determine the effectiveness of NPWT in the prevention of postoperative abdominal SWD in a predefined level of risk population. This level 1 study will provide further data for abdominal SWD risk classification, which is anticipated to inform preventive postoperative management. The study design uses a prospective real-world scenario in order to identify clinically significant differences between the intervention and control groups. TRIAL REGISTRATION: This trial was prospectively registered on 10 December 2012 with Australian and New Zealand Clinical Trials Network (ANZCTR): 12612001275853.

Compliance of post-radiation therapy head and neck cancer patients with caries preventive protocols.

BACKGROUND: Caries prevention is paramount in safeguarding the life quality of head and neck cancer patients and is dependent on patient compliance with caries preventive protocols. The purpose of this study was to examine this compliance. METHODS: All records of patients referred to one public oral medicine clinic servicing a head and neck oncology unit of one major Western Australian hospital, between January 2005 and December 2011, were examined. Data extracted included patient and cancer characteristics and compliance with dietary advice, dental care, oral hygiene instruction and fluoride use over a follow-up period of at least 12 months. Compliance was assessed against various oral health outcomes and patient characteristics. RESULTS: Of the 116 participants, 75.9% complied with all caries preventive measures over a mean follow-up period of 45 months. Non-compliance with regular dental attendance (P = 0.004), oral hygiene instruction (P = 0.009), dietary advice (P = 0.034) and daily fluoride use (P = 0.018) were associated with the development of dental caries post-treatment. The presence of dental caries at the time of cancer diagnosis was predictive of poorer compliance. CONCLUSIONS: High compliance with caries preventive measures is attainable in the head and neck cancer patient group. Factors other than fluoride use seem important in caries prevention.

An unusual case of Salmonella Enteritidis causing pneumonia, septic shock and multiple organ failure in an immunocompetent patient.

Salmonella species are facultative intracellular pathogens that most frequently cause self-limiting gastrointestinal disease, often acquired through the ingestion of contaminated food. We report the case of a 33-year-old otherwise healthy, not overtly immunosuppressed, man who was transferred to our facility with the chief complaint of respiratory failure and septic shock. Computed tomography of the chest revealed multifocal pneumonia in both lungs. A bronchial alveolar lavage was performed in the right middle lobe and cultures predominantly grew Salmonella enterica serovar Enteritidis. The patient received a prolonged course of antimicrobials, ultimately changing to oral levofloxacin. The etiology of the salmonella infection likely occurred through an aspiration event. Salmonella species are not a typical respiratory pathogen in immunocompetent hosts; however, clinicians should be aware of the possibility that salmonella species may be a pathogenic source of infection in the lungs; a prolonged course of antimicrobials may be warranted.

Evaluation of the first pharmacist-administered vaccinations in Western Australia: a mixed-methods study.

OBJECTIVES: This study evaluated the uptake of Western Australian (WA) pharmacist vaccination services, the profiles of consumers being vaccinated and the facilitators and challenges experienced by pharmacy staff in the preparation, implementation and delivery of services. DESIGN: Mixed-methods methodology with both quantitative and qualitative data through surveys, pharmacy computer records and immuniser pharmacist interviews. SETTING: Community pharmacies in WA that provided pharmacist vaccination services between March and October 2015. PARTICIPANTS: Immuniser pharmacists from 86 pharmacies completed baseline surveys and 78 completed exit surveys; computer records from 57 pharmacies; 25 immuniser pharmacists were interviewed. MAIN OUTCOME MEASURES: Pharmacy and immuniser pharmacist profiles; pharmacist vaccination services provided and consumer profiles who accessed services. RESULTS: 15 621 influenza vaccinations were administered by immuniser pharmacists at 76 WA community pharmacies between March and October 2015. There were no major adverse events, and <1% of consumers experienced minor events which were appropriately managed. Between 12% and 17% of consumers were eligible to receive free influenza vaccinations under the National Immunisation Program but chose to have it at a pharmacy. A high percentage of vaccinations was delivered in rural and regional areas indicating that provision of pharmacist vaccination services facilitated access for rural and remote consumers. Immuniser pharmacists reported feeling confident in providing vaccination services and were of the opinion that services should be expanded to other vaccinations. Pharmacists also reported significant professional satisfaction in providing the service. All participating pharmacies intended to continue providing influenza vaccinations in 2016. CONCLUSIONS: This initial evaluation of WA pharmacist vaccination services showed that vaccine delivery was safe. Convenience and accessibility were important aspects in usage of services. There is scope to expand pharmacist vaccination services to other vaccines and younger children; however, government funding to pharmacists needs to be considered.

A trial of a self-assessment tool of problems following treatment of colorectal cancer: a prospective study in Australia primary care.

Patients treated for colorectal cancer (CRC) experience considerable physical, social and psychological morbidity. In this study, 66 participants with stages I-III CRC were enrolled in this study. Participants completed the self-assessment tool for patients (SATp) over a 5-month period and visited a general practitioner with a copy of their SATp to assist in the management of any problems associated with CRC treatment. General practitioners' notes were reviewed for management actions. Of the 66 participants, 57 visited a general practitioner over the 5-month study period. A total of 547 problems were identified (median 7; IQR: 3-12.25). Participants with physical problems were more likely to consult their general practitioner (OR: 1.84, CI: 1.05-3.21, P = 0.03) compared to those with psychological problems. The number of problems experienced by participants did not have any influence on the decision to visit a general practitioner. Psychological problems (P < 0.01) significantly reduced over the 5-month study period. Regular use of the SATp facilitates the identification of long-term CRC treatment-related problems. Some of these problems could then be addressed in primary care.

Differences in the use of vision and proprioception for postural control in autism spectrum disorder.

BACKGROUND: People with autism spectrum disorders (ASDs) also have poorer fundamental motor skills. The development of postural control underlies both social and motor skills. All three elements are facilitated by the active use of visual information. This study compares how adults with ASD and typically developed adults (TDAs) respond to a postural illusion induced using neck vibration. Adults with ASD unlike the TDA, were not expected to correct the illusion using vision. METHODS: The study used intermittent (15off, 5on) posterior neck vibration during 200 s of quiet stance to induce a postural illusion. In TDAs and only in the absence of vision this protocol induces a forward body lean. Participants (12 ASD, 20 TDA) undertook four conditions combining vibration and visual occlusion. RESULTS: As predicted, TDA were only affected by the postural illusion when vision was occluded (vibration condition: vision occluded (n=1) p=0.0001; vision available (n=3) p>0.2466). Adults with ASD were affected by the postural illusion regardless of the availability of vision (all conditions p<0.0007). CONCLUSIONS: Our findings indicated the adults with ASD did not use visual information to control standing posture. In light of existing evidence that vision-for-perception is processed typically in ASD, our findings support a specific deficit in vision-for-action. These findings may explain why individuals with ASD experience difficulties with both social and motor skills since both require vision-for-action. Further research needs to investigate the division of these visual learning pathways in order to provide more specific intervention opportunities in ASD.

The effects of firocoxib on cautery disbudding pain and stress responses in preweaned dairy calves.

Perioperative analgesic effects of oral firocoxib following cautery disbudding were investigated in preweaned calves. Twenty Holstein calves approximately 4 to 6wk old received a single oral dose of firocoxib, a nonsteroidal antiinflammatory, at 0.5mg/kg (n=10) or placebo (n=10) in a randomized controlled clinical trial. Responses, including ocular temperature determined by infrared thermography, pressure algometry measuring mechanical nociception threshold, and heart rate, were evaluated at 2, 4, 7, 8, and 24h after cornual nerve block and cautery disbudding. Blood samples were collected over 96h and analyzed for plasma cortisol and substance P concentrations by RIA. Additionally, ex vivo prostaglandin E2 concentrations were determined over a 72-h study period using an enzyme immunoassay. Data were analyzed using a linear mixed effects model with repeated measures. An inhibition of ex vivo prostaglandin E2 synthesis was observed from 12 to 48h following disbudding in calves treated with firocoxib. Cautery disbudding was associated with an increased nociception for the duration of sampling (24h). During the initial 24-h period following disbudding, no difference in response between treatment groups was noted. Following 24h, mean cortisol concentrations diverged between the 2 study groups with placebo-treated calves having increased cortisol concentrations at approximately 48h after disbudding. Furthermore, the overall integrated cortisol response as calculated as area under the effect curve tended to be reduced in firocoxib-treated calves. The prolonged effects of cautery dehorning require further investigation. Moreover, the effect of firocoxib on cortisol reduction observed in this study requires additional exploration.

A new class of cancer-associated PTEN mutations defined by membrane translocation defects.

Phosphatase and tensin homolog (PTEN), which negatively regulates tumorigenic phosphatidylinositol (3,4,5)-trisphosphate (PIP3) signaling, is a commonly mutated tumor suppressor. The majority of cancer-associated PTEN mutations block its essential PIP3 phosphatase activity. However, there is a group of clinically identified PTEN mutations that maintain enzymatic activity, and it is unknown how these mutations contribute to tumor pathogenesis. Here, we show that these enzymatically competent PTEN mutants fail to translocate to the plasma membrane where PTEN converts PIP3 to PI(4,5)P2. Artificial membrane tethering of the PTEN mutants effectively restores tumor suppressor activity and represses excess PIP3 signaling in cells. Thus, our findings reveal a novel mechanism of tumorigenic PTEN deficiency.

Oral cavity squamous cell carcinoma - characteristics and survival in aboriginal and non-aboriginal Western australians.

BACKGROUND: Squamous cell carcinoma (SCC) is the most common type of malignancy affecting the oral cavity. While exposures to main risk factors for oral SCC such as smoking and alcohol use are higher amongst the Aboriginal people, little is known about oral cancer in this population. This study aimed to describe characteristics and survival of oral SCC in Aboriginal and non-Aboriginal Western Australians. METHODS: All primary oral SCC cases reported to the Western Australian Cancer Registry (WACR) between 1990 and 1999 were analysed with respect to person characteristics including: date of birth, sex and indigenous status; and disease characteristics including: date of biopsy, disease stage and site as well as date of recurrence and date of death. Exclusion criteria included diagnosis not based on incisional or excisional biopsy, diagnosis other than oral SCC or a history of another malignant neoplasm. RESULTS: Aboriginal individuals were more likely to reside in rural areas. No statistically significant differences in oral SCC characteristics and survival were noted between Aboriginal and non-Aboriginal Western Australians. CONCLUSION: This study provides new information on person and disease characteristics of Aboriginal Western Australians diagnosed with oral SCC.

Evaluating approaches to measuring ocular pain in bovine calves with corneal scarification and infectious bovine keratoconjunctivitis-associated corneal ulcerations.

Infectious bovine keratoconjunctivitis (IBK) is a common ocular disease in cattle, associated with a 6.8 to 13.6 kg decrease in weaning weight. Antibiotic therapy is available but it is unclear if pain mitigation as an adjunct therapy would reduce the weight loss associated with IBK. Before assessing the impact of pain mitigation therapies, it is first necessary to validate approaches to qualifying ocular pain. The objective of this study was to evaluate approaches to qualifying ocular pain in bovine calves (Bos taurus) with IBK. Our a priori assumption was that scarification or corneal ulcerations consistent with IBK are painful compared to normal eyes. To quantify this difference in pain, we assessed 4 tools: pressure algometry-mechanical nociceptive threshold (PA-MNT), corneal touch thresholds (CTT) obtained with the use of a Cochet-Bonnet aesthesiometer, and assessment for the presence of blepharospasm and photophobia as metrics for pain. Using a 1-eye randomized controlled challenge trial, 31 calves with healthy eyes were randomly allocated to treatment groups, and then a left or right eye was randomly assigned for corneal scarification and inoculation with Moraxella bovoculi or Moraxella bovis. A repeated measures analysis of variance was used for PA-MNT, with significance set at P < 0.05. A log (base 10) transformation was used to stabilize the variance, and Tukey's t tests were used to test differences between assessment days for each landmark. Calves had statistically significantly lower PA-MNT scores (which indicates more pain) the day after scarification relative to baseline measurements (4 d before scarification). For example, at 1 landmark the median PA-MNT (kg/force) prescarification was 4.82 (95% confidence interval [CI]: 3.92-5.93) and 3.43 (95% CI: 2.79-4.22) postscarification. These data suggest PA-MNT may be a tool for quantifying ocular pain in calves. No differences (P < 0.1) in PA-MNT scores between scarified and not-scarified eyes were detected for any landmark on any day. This result suggests that the pain response occurs over the entire face, not just the affected eye. Corneal ulcerations consistent with IBK were not associated with statistically significant differences in PA-MNT or CTT at eye or calf levels. Not surprisingly, scarified eyes were more likely to exhibit blepharospasm and photophobia compared to healthy eyes. Due to blepharospasm, the use of the Cochet-Bonnet to evaluate corneal sensitivity by CTT was of limited value.

Sustained reduction of alloantibody secreting plasma cells and donor specific antibody with proteasome inhibition in mice.

The long-lived plasma cells, which develop after alloantigen sensitization, produce donor specific alloantibodies (DSAs) that generate a positive serum cross-match and preclude transplantation. Bortezomib, a proteasome inhibitor, is being investigated in clinical desensitization protocols, however preclinical studies in a transplant model are nonexistent. We hypothesized that sustained treatment with only a proteasome inhibitor would eliminate plasma cells and reduce DSA over time. Cardiac allografts were transplanted into murine recipients. Eight weeks after allograft rejection the proteasome inhibitor, bortezomib, was injected intravenously twice weekly for 60 days. Serum alloantibody responses were assayed using flow cross-match. Total and alloreactive plasma cell numbers were enumerated using flow cytometry and ELISPOT. All recipients of cardiac allografts rejected their graft promptly within 16 days and demonstrated alloantibody by flow cross-match. DSA was sustained in the control mice while mice treated with bortezomib had sustained elimination of DSA and a marked reduction in plasma cell population. Also, bortezomib was associated with an increased level of BLyS. Within a murine model, proteasome inhibition can eliminate alloantibody secreting plasma cells, and reduce alloantibody. Cessation of bortezomib is not associated with return of DSA.

The economic value of organic dairy farms in Vermont and Minnesota.

This study quantifies the overall economic values of organic dairy farms in Vermont and Minnesota and estimates the economic impacts of organic dairy farm sales relative to an equivalent level of sales from conventional dairy farms in those states. This question is of interest because the development of the organic dairy sector has allowed some farms that likely would not have remained in the conventional dairy business to continue being economically viable as organic dairy farms. Thus, these sales provide an economic impact in regions when this milk is exported to nonproducing regions. Organic and conventional dairy farm financial data in Vermont and Minnesota were collected and assembled to develop dairy farm production functions by region and dairy type. These production functions were then used in state-level input-output models to calculate economic impacts. The opportunity costs of organic dairy farm production were also estimated by comparing the relative statewide economic impacts of organic and conventional dairy farms if both experience a hypothetical 5-million-dollar increase in sales. Between 2008 and 2010, Vermont's 180 organic dairy farms annually contributed $76.3 million in output (the value of an industry's production within the state), 808 jobs, $34.1 million in gross state product, and $26.3 million in labor income to Vermont's economy. Between 2009 and 2011, Minnesota's 114 organic dairy farms annually contributed $77.7 million in output, 552 jobs, $32.1 million in gross state product, and $21 million in labor income to Minnesota's economy. In Vermont, organic dairy farm sales revenue would result in greater state-wide impacts of 3% in output, 39% in labor income, 33% in gross state product, and 46% in employment relative to the impacts from an equivalent level of sales revenue to conventional dairy farms. In Minnesota, these economic impacts are 4, 9, 11, and 12% greater, respectively, for organic dairy farms relative to conventional dairy farms. This study concludes that organic dairy farms may contribute more to the local economy than average and similar-size conventional dairy farms in the Northeast and Upper Midwest and that organic dairy farm milk production supports economic development in rural communities.

Nicotinamide N-methyltransferase expression in SH-SY5Y neuroblastoma and N27 mesencephalic neurones induces changes in cell morphology via ephrin-B2 and Akt signalling.

Nicotinamide N-methyltransferase (NNMT, E.C. 2.1.1.1) N-methylates nicotinamide to produce 1-methylnicotinamide (MeN). We have previously shown that NNMT expression protected against neurotoxin-mediated cell death by increasing Complex I (CxI) activity, resulting in increased ATP synthesis. This was mediated via protection of the NDUFS3 subunit of CxI from degradation by increased MeN production. In the present study, we have investigated the effects of NNMT expression on neurone morphology and differentiation. Expression of NNMT in SH-SY5Y human neuroblastoma and N27 rat mesencephalic dopaminergic neurones increased neurite branching, synaptophysin expression and dopamine accumulation and release. siRNA gene silencing of ephrin B2 (EFNB2), and inhibition of Akt phosphorylation using LY294002, demonstrated that their sequential activation was responsible for the increases observed. Incubation of SH-SY5Y with increasing concentrations of MeN also increased neurite branching, suggesting that the effects of NNMT may be mediated by MeN. NNMT had no significant effect on the expression of phenotypic and post-mitotic markers, suggesting that NNMT is not involved in determining phenotypic fate or differentiation status. These results demonstrate that NNMT expression regulates neurone morphology in vitro via the sequential activation of the EFNB2 and Akt cellular signalling pathways.

Economic feasibility of converting cow manure to electricity: a case study of the CVPS Cow Power program in Vermont.

A case study of the Central Vermont Public Service Corporation (CVPS) Cow Power program examines the economic feasibility for dairy farms to convert cow manure into electricity via anaerobic methane digestion. The study reviews the mechanism for CVPS, dairy farms, electricity customers, and government agencies to develop and operate the program since 2004, examines the costs and returns for the participating dairy farms, and assesses their cash flow over a period of 7 yr under different scenarios. With 6 dairy farms generating about 12 million kilowatt-hours of electricity per year and more than 4,600 CVPS electricity customers voluntarily paying premiums of $0.04 per kilowatt-hour, or a total of about $470,000 per year, the CVPS Cow Power program represents a successful and locally sourced renewable energy project with many environmental and economic benefits. Factors for the successful development and operation of the program include significant grants from government agencies and other organizations, strong consumer support, timely adjustments to the basic electricity price paid to the farms, and close collaboration among the participating parties. This study confirms that it is technically feasible to convert cow manure to electricity on farms, but the economic returns depend highly on the base electricity price, premium rate, financial supports from government agencies and other organizations, and sales of the byproducts of methane generation.

Consolidation and reconsolidation of contextual fear memory requires mammalian target of rapamycin-dependent translation in the dorsal hippocampus.

The mammalian target of rapamycin (mTOR) pathway is important for regulating protein translation. The present study characterized the role of mTOR-dependent translation in the dorsal hippocampus (DH) during the consolidation and reconsolidation of contextual fear memory. We first showed that fear conditioning resulted in increased phosphorylation of p70s6 kinase (p70s6K) in the DH and that infusion of the mTOR inhibitor rapamycin (RAP) into the DH immediately after training disrupted formation of long-term contextual fear memory. Additionally we showed that p70s6K was activated after retrieval of a previously stored fear memory, and inhibition of mTOR by DH infusion of RAP blocked the reconsolidation of contextual fear memory. Together these results demonstrate that within the DH translational control through the mTOR pathway is important for consolidation as well as the stability of fear memory after retrieval.