Defective Allele of the Neuronal Nitric Oxide Synthase Gene Increases Insulin Resistance During Acute Phase of Myocardial Infarction.

BACKGROUND: Glycemic disorders are strong predictors of mortality in ST-elevation myocardial infarction (STEMI) patients, and disruption in nitric oxide (NO) production is associated with insulin-resistant states. We evaluated whether a defective allele of the neuronal nitric oxide synthase (nNOS) gene (NOS1) might influence insulin response and blood-glucose balance during the acute phase of STEMI and if post-infarction total plasma-NO levels and vasodilation scores varied across nNOS genotypes. METHODS: Consecutive patients with STEMI (n=354) underwent clinical evaluations and genotyping for the promoter variation rs41279104. In-hospital clinical and blood evaluations were performed at admission and five days after STEMI, with glycemic, insulinemic, and disposition indices assessed at the same times. Flow-mediated dilation (FMD) was assessed by reactive hyperemia on the 30th day. RESULTS: Homozygotes for the defective allele (A) showed lower glycemia and insulin sensitivity on day 1 while showing the highest ß-cell function and no changes in the circulating NO pool, which is compatible with hyperresponsive ß cells counteracting the inherent glucose-resistant state of AA patients. At day 5, glycemic scores had shifted to indicate greater insulin sensitivity among A homozygotes, paralleled by a significant yet poor increase in NO bioavailability compared to that among G carriers. All in all, defective homozygotes showed greater insulin resistance at admission that had reversed by 5 days after STEMI. Even so, A carriers developed lower FMD scores compared to G homozygotes after the acute phase. CONCLUSION: A defective nNOS allele (and due decline in NO production) seemed to elicit a hyperinsulinemia response to compensate for an insulin-resistant state during the acute phase of STEMI and to be associated with poor endothelial function after the acute phase.

Case report on metastatic pelvic bone tumor treated with frozen autograft by liquid nitrogen.

INTRODUCTION AND IMPORTANCE: Frozen autograft recycling has been used for biological reconstruction of bone defects following tumor excision, more commonly in extremities. We report on the histological outcome of a pelvic recycled frozen autograft. CASE PRESENTATION: We investigated the pelvic frozen autograft removed in 2 years and 8 months after surgery because of soft tissue recurrence in pelvic floor. The autograft bone showed no evidence of revitalization and was non-viable with patchy inflammation, and no residual tumor. There was only fibrous union but the autograft bone remained mechanically stable. CLINICAL DISCUSSION: We confirmed the clearance of tumor cells with the treatment with liquid nitrogen. The union at the host-graft junction might be affected by the previous radiotherapy, the presence of infection, the small contact area limited by the anatomy, and the inadequate compression across the osteotomy interface with the fixation. CONCLUSION: Frozen autograft treated by liquid nitrogen can be used safely for biological reconstructions after pelvic tumor excision.

Experiences of adolescent lung transplant recipients: A qualitative study.

Many young transplant recipients experience psychological distress and adjustment difficulties, yet there is little research investigating lung transplantation from the recipients' perspective. This qualitative study aimed to explore experiences of young people who underwent lung transplantation. Semi-structured interviews were conducted with six lung transplant recipients (aged 15-18). Interviews were analysed using IPA, a qualitative research approach examining how people make sense of their major life experiences. The analysis revealed three master themes: "Living with Dodgy Lungs" outlined how participants dealt with their experiences, managing through accepting or discussing their feelings with others, although talking was often difficult. "The Big Deal" reflected participants' experiences of the process, their expectations, and the contrast of their lives pre- and post-transplant. Inherent in their accounts was the profound meaning ascribed to transplantation, the emotional turmoil, and impact on their lives. "A Sense of Self" illustrated participants' developing identities within their social contexts and at times isolating experiences. The results highlight key areas where adolescent lung transplant recipients could be supported by clinicians, enabling the promotion of psychological well-being. Examples include supporting identity integration post-transplant, facilitating social inclusion, considering alternative means of support, and involving adolescents in healthcare decisions.

A highly prevalent equine glycogen storage disease is explained by constitutive activation of a mutant glycogen synthase.

BACKGROUND: Equine type 1 polysaccharide storage myopathy (PSSM1) is associated with a missense mutation (R309H) in the glycogen synthase (GYS1) gene, enhanced glycogen synthase (GS) activity and excessive glycogen and amylopectate inclusions in muscle. METHODS: Equine muscle biochemical and recombinant enzyme kinetic assays in vitro and homology modelling in silico, were used to investigate the hypothesis that higher GS activity in affected horse muscle is caused by higher GS expression, dysregulation, or constitutive activation via a conformational change. RESULTS: PSSM1-affected horse muscle had significantly higher glycogen content than control horse muscle despite no difference in GS expression. GS activity was significantly higher in muscle from homozygous mutants than from heterozygote and control horses, in the absence and presence of the allosteric regulator, glucose 6 phosphate (G6P). Muscle from homozygous mutant horses also had significantly increased GS phosphorylation at sites 2+2a and significantly higher AMPKα1 (an upstream kinase) expression than controls, likely reflecting a physiological attempt to reduce GS enzyme activity. Recombinant mutant GS was highly active with a considerably lower Km for UDP-glucose, in the presence and absence of G6P, when compared to wild type GS, and despite its phosphorylation. CONCLUSIONS: Elevated activity of the mutant enzyme is associated with ineffective regulation via phosphorylation rendering it constitutively active. Modelling suggested that the mutation disrupts a salt bridge that normally stabilises the basal state, shifting the equilibrium to the enzyme's active state. GENERAL SIGNIFICANCE: This study explains the gain of function pathogenesis in this highly prevalent polyglucosan myopathy.

Fatal bilateral lower-limb deep vein thrombosis and pulmonary embolism following single digit replantation.

Venous thromboembolism in hand surgery is rare. There is no report in the literature on postoperative mortality from venous thromboembolism following microsurgery in upper limbs. We report the case of a 56-year-old Chinese man who died from pulmonary embolism as a result of bilateral lower-limb deep vein thrombosis following prolonged surgery under general anaesthesia after replantation of a finger. This case raises awareness of the need for precautions against venous thromboembolism following prolonged microsurgery and identification of high-risk patients.

Anxiolytic-like and anxiogenic-like effects of nicotine are regulated via diverse action at ß2*nicotinic acetylcholine receptors.

BACKGROUND AND PURPOSE: Nicotine dose-dependently activates or preferentially desensitizes ß2 subunit containing nicotinic ACh receptors (ß2*nAChRs). Genetic and pharmacological manipulations assessed effects of stimulation versus inhibition of ß2*nAChRs on nicotine-associated anxiety-like phenotype. EXPERIMENTAL APPROACH: Using a range of doses of nicotine in ß2*nAChR subunit null mutant mice (ß2KO; backcrossed to C57BL/6J) and their wild-type (WT) littermates, administration of the selective ß2*nAChR agonist, 5I-A85380, and the selective ß2*nAChR antagonist dihydro-ß-erythroidine (DHßE), we determined the behavioural effects of stimulation and inhibition of ß2*nAChRs in the light-dark and elevated plus maze (EPM) assays. KEY RESULTS: Low-dose i.p. nicotine (0.05 mg·kg(-) 1) supported anxiolysis-like behaviour independent of genotype whereas the highest dose (0.5 mg·kg(-1) ) promoted anxiogenic-like phenotype in WT mice, but was blunted in ß2KO mice for the measure of latency. Administration of 5I-A85380 had similar dose-dependent effects in C57BL/6J WT mice; 0.001 mg·kg(-1) 5I-A85380 reduced anxiety on an EPM, whereas 0.032 mg·kg(-1) 5I-A85380 promoted anxiogenic-like behaviour in both the light-dark and EPM assays. DHßE pretreatment blocked anxiogenic-like effects of 0.5 mg·kg(-1) nicotine. Similarly to DHßE, pretreatment with low-dose 0.05 mg·kg(-1) nicotine did not accumulate with 0.5 mg·kg(-1) nicotine, but rather blocked anxiogenic-like effects of high-dose nicotine in the light-dark and EPM assays. CONCLUSIONS AND IMPLICATIONS: These studies provide direct evidence that low-dose nicotine inhibits nAChRs and demonstrate that inhibition or stimulation of ß2*nAChRs supports the corresponding anxiolytic-like or anxiogenic-like effects of nicotine. Inhibition of ß2*nAChRs may relieve anxiety in smokers and non-smokers alike.

Coat color genotypes and risk and severity of melanoma in gray quarter horses.

BACKGROUND: Both graying and melanoma formation in horses have recently been linked to a duplication in the STX17 gene. This duplication, as well as a mutation in the ASIP gene that increases MC1R pathway signaling, affects melanoma risk and severity in gray horses. OBJECTIVE: To determine if melanoma susceptibility in gray Quarter Horses (QH) is lower than gray horses from other breeds because of decreased MC1R signaling resulting from a high incidence of the MC1R chestnut coat color allele in the QH population. ANIMALS: A total of 335 gray QH with and without dermal melanomas. METHODS: Blood or hair root samples were collected from all horses for DNA extraction and genotyping for STX17, ASIP, and MC1R genotypes. Age, sex, and external melanoma presence and grade were recorded. The effect of age and genotype on melanoma presence and severity was evaluated by candidate gene association. RESULTS: Melanoma prevalence (16%) and grade (0.35) in this QH cohort was lower than that reported in other breeds. Age was significantly associated with melanoma prevalence (P = 5.28 × 10(-11)) and severity (P = 2.2 × 10(-13)). No significant effect of MC1R genotype on melanoma prevalence or severity was identified. An effect of ASIP genotype on melanoma risk was not detected. Low STX17 homozygosity precluded evaluation of the gray allele effect. CONCLUSION AND CLINICAL IMPORTANCE: Melanoma prevalence and severity is lower in this population of gray QH than what is reported in other breeds. This could be because of the infrequent STX17 homozygosity, a mitigating effect of the MC1R mutation on ASIP potentiation of melanoma, other genes in the MC1R signaling pathway, or differences in breed genetic background.

Muscle microvascular recruitment predicts insulin sensitivity in middle-aged patients with type 1 diabetes mellitus.

AIMS/HYPOTHESIS: Insulin delivery to muscle is rate-limiting for insulin's metabolic action and is regulated by insulin's own action to increase skeletal muscle blood flow and to recruit microvasculature. Microvascular dysfunction has been observed in insulin resistant states. We investigated the relation between insulin's action to recruit microvasculature and its metabolic action in type 1 diabetes. METHODS: Near euglycaemia was obtained by an overnight insulin infusion during 17 inpatient admissions of participants with type 1 diabetes. This was followed by a 2 h 1 mU kg⁻¹ min⁻¹ euglycaemic-hyperinsulinaemic clamp. Microvascular blood volume (MBV) was assessed using contrast-enhanced ultrasound 10 min before and 30 min after starting the clamp. RESULTS: We observed that, after overnight modest hyperinsulinaemia (average ≈ 286 pmol/l), MBV was positively related to the steady-state insulin sensitivity measured during the subsequent clamp (r = 0.62, p = 0.008). The more marked hyperinsulinaemia during the clamp (average steady-state insulin ≈ 900 pmol/l) increased MBV in the more insulin resistant participants within 30 min but not in the insulin sensitive participants. The change in MBV during the clamp was negatively correlated to the insulin sensitivity (r = -0.55, p = 0.022). As a result, MBV after 30 min of marked hyperinsulinaemia was comparable between the insulin sensitive and resistant participants. CONCLUSIONS/INTERPRETATION: We conclude that moderate overnight hyperinsulinaemia recruited microvasculature in the more sensitive participants, while higher levels of plasma insulin were needed for more insulin resistant participants. This suggests that microvascular responsiveness to insulin is one determinant of metabolic insulin sensitivity in type 1 diabetes.

The effect of dietary protein level on performance characteristics of coccidiosis vaccinated and nonvaccinated broilers following mixed-species Eimeria challenge.

A series of experiments were conducted to investigate the effect of starter diet protein levels on the performance of broilers vaccinated with a commercially available live oocyst coccidiosis vaccine before subsequent challenge with a mixed-species Eimeria challenge. Data indicated that an increasing protein concentration in the starter diet improved broiler performance during coccidiosis vaccination. Prechallenge performance data indicated that vaccination could decrease BW and increase feed conversion ratio. The time period most important for the observed effects appeared to be between 13 and 17 d of age. This reduction in performance parameters of vaccinated broilers compared with nonvaccinated broilers was eliminated by the conclusion of the experiments (27 d) in the diet groups with higher protein. Vaccination was effective at generating protective immunity against Eimeria challenge, as evidenced by increased (P < 0.05) BW gain, improved feed conversion, reduced postchallenge mortality, and reduced lesion development in vaccinated broilers compared with nonvaccinated broilers. These observations support numerous other reports that confirm live oocyst vaccination can be used effectively as a preventive against avian coccidiosis in commercially reared broilers. More important, these findings suggest that reduced protein concentration of starter diets can lead to significant losses in broiler performance when using a vaccination program to prevent coccidiosis.

Loss of PKCδ results in characteristics of Sjögren's syndrome including salivary gland dysfunction.

OBJECTIVES: Chronic infiltration of lymphocytes into the salivary and lacrimal glands of patients with Sjögren's syndrome (SS) leads to destruction of acinar cells and loss of exocrine function. Protein kinase C-delta (PKCδ) is known to play a critical role in B-cell maintenance. Mice in which the PKCδ gene has been disrupted have a loss of B-cell tolerance, multiple organ lymphocytic infiltration, and altered apoptosis. To determine whether PKCδ contributes to the pathogenesis of SS, we quantified changes in indicators of SS in PKCδ-/- mice as a function of age. Salivary gland histology, function, the presence of autoantibodies, and cytokine expression were examined. MATERIALS AND METHODS: Submandibular glands were examined for the presence of lymphocytic infiltrates, and the type of infiltrating lymphocyte and cytokine deposition was evaluated by immunohistochemistry. Serum samples were tested by autoantibody screening, which was graded by its staining pattern and intensity. Salivary gland function was determined by saliva collection at various ages. RESULTS: PKCδ-/- mice have reduced salivary gland function, B220+ B-cell infiltration, anti-nuclear antibody production, and elevated IFN-γ in the salivary glands as compared to PKCδ+/+ littermates. CONCLUSIONS: PKCδ-/- mice have exocrine gland tissue damage indicative of a SS-like phenotype.

Rechargeable wireless EMG sensor for prosthetic control.

Surface electrodes in modern myoelectric prosthetics are often embedded in the prosthesis socket and make contact with the skin. These electrodes detect and amplify muscle action potentials from voluntary contractions of the muscle in the residual limb and are used to control the prosthetic's movement and function. There are a number of performance-related deficiencies associated with external electrodes including the maintenance of sufficient electromyogram (EMG) signal amplitude, extraneous noise acquisition, and proper electrode interface maintenance that are expected to be improved or eliminated using the proposed implanted sensors. This research seeks to investigate the design components for replacing external electrodes with fully-implantable myoelectric sensors that include a wireless interface to the prosthetic limbs. This implanted technology will allow prosthetic limb manufacturers to provide products with increased performance, capability, and patient-comfort. The EMG signals from the intramuscular recording electrode are amplified and wirelessly transmitted to a receiver in the prosthetic limb. Power to the implant is maintained using a rechargeable battery and an inductive energy transfer link from the prosthetic. A full experimental system was developed to demonstrate that a wireless biopotential sensor can be designed that meets the requirements of size, power, and performance for implantation.

Estimated prevalence of the Type 1 Polysaccharide Storage Myopathy mutation in selected North American and European breeds.

The GYS1 gene mutation that is causative of Type 1 Polysaccharide Storage Myopathy (PSSM) has been identified in more than 20 breeds of horses. However, the GYS1 mutation frequency or Type 1 PSSM prevalence within any given breed is unknown. The purpose of this study was to determine the frequency of the GYS1 mutation and prevalence of genetic susceptibility to Type 1 PSSM in selected breeds from Europe and North America. The GYS1 mutation was detected in 11 breeds, including, in order of increasing allele frequency, Shires, Morgans, Appaloosas, Quarter Horses, Paints, Exmoor Ponies, Saxon-Thuringian Coldbloods, South German Coldbloods, Belgians, Rhenish German Coldbloods and Percherons. The prevalence of genetic susceptibility to Type 1 PSSM in these breeds varied from 0.5% to 62.4%. The GYS1 mutation was not found in the sampled Thoroughbreds, Akhal-Tekes, Connemaras, Clydesdales, Norwegian Fjords, Welsh Ponies, Icelandics, Schleswig Coldbloods or Hanoverians, but failure to detect the mutation does not guarantee its absence. This knowledge will help breed associations determine whether they should screen for the GYS1 mutation and will alert veterinarians to a possible differential diagnosis for muscle pain, rhabdomyolysis or gait abnormalities.

Presence of the glycogen synthase 1 (GYS1) mutation causing type 1 polysaccharide storage myopathy in continental European draught horse breeds.

The purpose of this study was to determine which continental European draught horse breeds harbour a mutation in the glycogen synthase 1 gene (GYS1) that is known to be responsible for type 1 polysaccharide storage myopathy in quarter horses and North American draught horses. Of a non-random selection of continental European draught horses belonging to 13 breeds, 62 per cent (250 of 403) tested were found to carry the mutant allele. The horses were located in Belgium, France, Germany, The Netherlands, Spain and Sweden. The mutation was identified in animals from each of the breeds examined. In the breeds in which more than 15 animals were available for testing, the highest percentages of GYS1-positive horses were found in the Belgian trekpaard (92 per cent; 35 of 38 horses tested), Comtois (80 per cent; 70 of 88), Netherlands trekpaard (74 per cent; 17 of 23), Rheinisch-Deutsches kaltblut (68 per cent; 30 of 44) and Breton (64 per cent; 32 of 51).

Loss of protein kinase C delta alters mammary gland development and apoptosis.

As apoptotic pathways are commonly deregulated in breast cancer, exploring how mammary gland cell death is regulated is critical for understanding human disease. We show that primary mammary epithelial cells from protein kinase C delta (PKCδ) -/- mice have a suppressed response to apoptotic agents in vitro. In the mammary gland in vivo, apoptosis is critical for ductal morphogenesis during puberty and involution following lactation. We have explored mammary gland development in the PKCδ -/- mouse during these two critical windows. Branching morphogenesis was altered in 4- to 6-week-old PKCδ -/- mice as indicated by reduced ductal branching; however, apoptosis and proliferation in the terminal end buds was unaltered. Conversely, activation of caspase-3 during involution was delayed in PKCδ -/- mice, but involution proceeded normally. The thymus also undergoes apoptosis in response to physiological signals. A dramatic suppression of caspase-3 activation was observed in the thymus of PKCδ -/- mice treated with irradiation, but not mice treated with dexamethasone, suggesting that there are both target- and tissue-dependent differences in the execution of apoptotic pathways in vivo. These findings highlight a role for PKCδ in both apoptotic and nonapoptotic processes in the mammary gland and underscore the redundancy of apoptotic pathways in vivo.

System to improve AED resuscitation using interactive CPR coaching.

A positive impact on cardiac arrest survival has been demonstrated with the substantial reduction in time to defibrillation provided by the widespread deployment of automated external defibrillators (AEDs). However, recent studies have identified the importance of performing chest compressions before defibrillation in facilitating effective recovery from long duration ventricular fibrillation (VF). Despite the importance of cardiopulmonary resuscitation (CPR), effective performance of it in the field is hampered by many problems including the dependence on rescuer technique, which is known to be variable even with trained professionals. This research seeks to enhance survival outcomes following resuscitation. A full experimental system was developed that used an instrumented CPR manikin to provide interactive CPR coaching while collecting performance data. This system was utilized in a controlled human CPR performance study comparing the differences in chest compression performance with and without visual coaching and with and without interactive performance feedback coaching. Results from the human study support a number of conclusions and recommendations. In general using any type of coaching provided improvements in all of the CPR performance measures excluding chest recoil where there was a slight decrease in performance. The statistical results also indicated that the audio/visual coaching conditions provided a more effective coaching condition with respect to chest compression rate. Most notably, the feedback conditions both provided a statistically significant or trends toward improving chest compression effectiveness and produced superior performance as a whole.

Cocaine-induced alterations in dopamine receptor signaling: implications for reinforcement and reinstatement.

The transition from casual drug use to addiction, and the intense drug craving that accompanies it, has been postulated to result from neuroadaptations within the limbic system caused by repeated drug exposure. This review will examine the implications of cocaine-induced alterations in mesolimbic dopamine receptor signaling within the context of several widely used animal models of addiction. Extensive evidence indicates that dopaminergic mechanisms critically mediate behavioral sensitization to cocaine, cocaine-induced conditioned place preference, cocaine self-administration, and the drug prime-induced reinstatement of cocaine-seeking behavior. The propagation of the long-term neuronal changes associated with recurring cocaine use appears to occur at the level of postreceptor signal transduction. Repeated cocaine treatment causes an up-regulation of the 3',5'-cyclic adenosine monophosphate (cAMP)-signaling pathway within the nucleus accumbens, resulting in a dys-regulation of balanced D1/D2 dopamine-like receptor signaling. The intracellular events arising from enhanced D1-like postsynaptic signaling mediate both facilitatory and compensatory responses to the further reinforcing effects of cocaine.

N-ViroTech--a novel process for the treatment of nutrient limited wastewaters.

As pulp and paper wastewaters are mostly deficient in nitrogen and phosphorus, historical practice has dictated that they cannot be effectively treated using microbiological processes without the addition of supplementary nutrients, such as urea and phosphoric acid. Supplementation is a difficult step to manage efficiently, requiring extensive post-treatment monitoring and some degree of overdosing to ensure sufficient nutrient availability under all conditions. As a result, treated wastewaters usually contain excess amounts of both nutrients, leading to potential impacts on the receiving waters such as eutrophication. N-ViroTech is a highly effective alternative treatment technology which overcomes this nutrient deficiency/excess paradox. The process relies on communities of nitrogen-fixing bacteria, which are able to directly fix nitrogen from the atmosphere, thus satisfying their cellular nitrogen requirements. The process relies on manipulation of growth conditions within the biological system to maintain a nitrogen-fixing population whilst achieving target wastewater treatment performance. The technology has significant advantages over conventional activated sludge operation, including: Improved environmental performance. Nutrient loadings in the final treated effluent for selected nitrogen and phosphorus species (particularly ammonium and orthophosphate) may be reduced by over 90% compared to conventional systems; Elimination of nitrogen supplementation, and minimisation of phosphorus supplementation, thus achieving significant chemical savings and resulting in between 25% and 35% savings in operational costs for a typical system; Self-regulation of nutrient requirements, as the bacteria only use as much nitrogen as they require, allowing for substantially less operator intervention and monitoring. This paper will summarise critical performance outcomes of the N-ViroTech process utilising results from laboratory-, pilot-scale and recent alpha-adopter, full-scale trials.

CT colonography: determination of optimal CT technique using a novel colon phantom.

The aim of this study was to determine the thickest slice at the lowest radiation dose for detection of colon polyps larger than 5mm in diameter at computed tomographic (CT) colonography. A colon phantom containing haustral folds, flexures, and straight segments was constructed of borosilicate. One hundred forty simulated polyps (5, 7, 10, and 12 mm) of various shapes (sessile, flat, and pedunculated) were attached at different colon locations (wall, base of fold, on the fold and fold tip). Polyps were positioned parallel, perpendicular, and oblique to the CT gantry. The air-filled phantom was scanned at different slice thicknesses (1.25-5 mm) and x-ray tube currents (5-308 mA). All polyps were identified in all data sets except one (1.25 mm slice thickness, 5 mA). In this acquisition, image noise reduced polyp visibility, and five of 140 (3%) polyps could not be identified. Unidentified polyps were 5 mm, flat or sessile in shape, located on the colon wall or base of the fold, and oblique or parallel to CT gantry. All tested CT techniques provided optimal polyp detection except settings at 1.25 mm and 5 mAs. Thin collimation (<5 mm) scans may not be necessary to detect clinically significant polyps.