HX600, a synthetic agonist for RXR-Nurr1 heterodimer complex, prevents ischemia-induced neuronal damage.

Ischemic stroke is amongst the leading causes of death and disabilities. The available treatments are suitable for only a fraction of patients and thus novel therapies are urgently needed. Blockage of one of the cerebral arteries leads to massive and persisting inflammatory reaction contributing to the nearby neuronal damage. Targeting the detrimental pathways of neuroinflammation has been suggested to be beneficial in conditions of ischemic stroke. Nuclear receptor 4A-family (NR4A) member Nurr1 has been shown to be a potent modulator of harmful inflammatory reactions, yet the role of Nurr1 in cerebral stroke remains unknown. Here we show for the first time that an agonist for the dimeric transcription factor Nurr1/retinoid X receptor (RXR), HX600, reduces microglia expressed proinflammatory mediators and prevents inflammation induced neuronal death in in vitro co-culture model of neurons and microglia. Importantly, HX600 was protective in a mouse model of permanent middle cerebral artery occlusion and alleviated the stroke induced motor deficits. Along with the anti-inflammatory capacity of HX600 in vitro, treatment of ischemic mice with HX600 reduced ischemia induced Iba-1, p38 and TREM2 immunoreactivities, protected endogenous microglia from ischemia induced death and prevented leukocyte infiltration. These anti-inflammatory functions were associated with reduced levels of brain lysophosphatidylcholines (lysoPCs) and acylcarnitines, metabolites related to proinflammatory events. These data demonstrate that HX600 driven Nurr1 activation is beneficial in ischemic stroke and propose that targeting Nurr1 is a novel candidate for conditions involving neuroinflammatory component.

Neuronally-directed effects of RXR activation in a mouse model of Alzheimer's disease.

Alzheimer's disease (AD) is characterized by extensive neuron loss that accompanies profound impairments in memory and cognition. We examined the neuronally directed effects of the retinoid X receptor agonist bexarotene in an aggressive model of AD. We report that a two week treatment of 3.5 month old 5XFAD mice with bexarotene resulted in the clearance of intraneuronal amyloid deposits. Importantly, neuronal loss was attenuated by 44% in the subiculum in mice 4 months of age and 18% in layer V of the cortex in mice 8 months of age. Moreover, bexarotene treatment improved remote memory stabilization in fear conditioned mice and improved olfactory cross habituation. These improvements in neuron viability and function were correlated with significant increases in the levels of post-synaptic marker PSD95 and the pre-synaptic marker synaptophysin. Moreover, bexarotene pretreatment improved neuron survival in primary 5XFAD neurons in vitro in response to glutamate-induced excitotoxicity. The salutary effects of bexarotene were accompanied by reduced plaque burden, decreased astrogliosis, and suppression of inflammatory gene expression. Collectively, these data provide evidence that bexarotene treatment reduced neuron loss, elevated levels of markers of synaptic integrity that was linked to improved cognition and in an aggressive model of AD.

Age-related decrease in stimulated glutamate release and vesicular glutamate transporters in APP/PS1 transgenic and wild-type mice.

We assessed baseline and KCl-stimulated glutamate release by using microdialysis in freely moving young adult (7 months) and middle-aged (17 months) transgenic mice carrying mutated human amyloid precursor protein and presenilin genes (APdE9 mice) and their wild-type littermates. In addition, we assessed the age-related development of amyloid pathology and spatial memory impaired in the water maze and changes in glutamate transporters. APdE9 mice showed gradual spatial memory impairment between 6 and 15 months of age. The stimulated glutamate release declined very robustly in 17-month-old APdE9 mice as compared to 7-month-old APdE9 mice. This age-dependent decrease in stimulated glutamate release was also evident in wild-type mice, although it was not as robust as in APdE9 mice. When compared to individual baselines, all aged wild-type mice showed 25% or greater increase in glutamate release upon KCl stimulation, but none of the aged APdE9 mice. There was an age-dependent decline in VGLUT1 levels, but not in the levels of VGLUT2, GLT-1 or synaptophysin. Astrocyte activation as measured by glial acidic fibrillary protein was increased in middle-aged APdE9 mice. Blunted pre-synaptic glutamate response may contribute to memory deficit in middle-aged APdE9 mice.

An implementation of the HACCP system in the production of food-packaging material.

The Hazard Analysis Critical Control Point (HACCP) system according to the Codex Alimentarius model was applied to the processes of five paper and paperboard mills and four plants further converting paper or board intended for contact with foodstuffs. The generalised flow diagrams of the processes are presented. Each of the overall processes contained 40-150 process steps. Normally three to five sessions with HACCP teams and additional private negotiations were needed for each mill or plant. Hazards leading to critical control points (CCPs) were microbiological (handling/storage, circulation water, starch, process environment) and physical (process environment) in mills, and microbiological (storage, lacquers or glues, packaging and process environment), chemical (printing) and physical (storage of products, packaging and process environment) in plants. Specifications, critical limits (e.g., based on different kinds of reports and instructions), monitoring methods (microbiological and visual) and frequency, responsibilities and corrective actions of the processes are presented. Most of the improvements focused on improving the process environment. In five cases, hygiene training was included in the implementation of the HACCP system.

Active and intelligent packaging: applications and regulatory aspects.

Changes in the way foods are produced, distributed, stored and retailed, reflecting the continuing increase in consumer demands for improved quality and extended shelf-life for packaged foods, are placing ever-greater demands on the performance of food packaging. Consumers want to be assured that the packaging is fulfilling its function of protecting the integrity, quality, freshness and safety of foods. To provide this assurance and help improve the performance of the packaging, innovative active and intelligent packaging concepts are being developed and applied in various countries. In Europe, however, the development and application of active and intelligent packaging systems have been limited thus far. The main reasons are legislative restrictions and a lack of knowledge about consumer acceptance, the efficacy of such systems, and the economic and environmental impact they may have. Therefore, in 1999, a European study was started within the framework of the EU FAIR R&D programme. It aims to initiate amendments to European legislation for food-contact materials to establish and implement active and intelligent systems within the current relevant regulations for packaged food in Europe. This paper presents an overview of existing active and intelligent systems and their current and future food-related applications. In addition, developments and trends in active and intelligent food packaging are discussed. The objectives and the work programme of the European project are reviewed and the results obtained so far are presented. The benefits for both the European consumer and the European food and food-packaging industries are highlighted.

Long-term follow-up of morbidity and mortality after aortic valve replacement with a mechanical valve prosthesis.

AIMS: The aim of this study was to determine the incidence of valve-related complications in patients with a mechanical aortic valve prosthesis and to identify risk factors for an adverse outcome. METHODS AND RESULTS: In the 424 patients, event-free survival rates 5 and 10 years after aortic valve replacement were 62% and 37%, respectively. The linearized incidence of thromboembolic events was 4.4% per patient-year, and of anticoagulant-related haemorrhage 8.5% per patient-year. Advanced NYHA functional class, atrial fibrillation, pure aortic regurgitation and thromboembolism prior to surgery decreased event-free survival. A history of pre-operative thromboembolism increased the risk for a first embolic event after aortic valve replacement (relative hazard [RH] 3.2), but was even more strongly associated with the risk for repeated events (> or =2 events, RH 5.4). After each thromboembolic episode that occurred, the risk for a subsequent one was increased. The risk for at least one, and up to three or more haemorrhages was increased in patients with a pre-operative history of bleeding (RH 3.3-5.1) and of atrial fibrillation (RH 1.8-3.9). The risk for a subsequent event was increased by a history of repeated haemorrhages, a short interval since previous bleeding, and high age. CONCLUSIONS: There were few factors strongly related to valve related morbidity. However, previous bleedings and previous thromboembolism were powerful risk factors for repeated events.

Low-energy epicardial pacing in children: the benefit of autocapture.

BACKGROUND: Permanent cardiac pacing in children results commonly in augmented energy consumption because of the high pacing rates and the ample stimulation safety margin applied in children. Cardiovascular anatomy and limited venous access sometimes preclude the otherwise preferred endocardial approach. In this multicenter patient series, we studied the feasibility, safety, and energy saving obtained by a combination of steroid-eluting epicardial leads with autocapture devices capable of ongoing adjustment of the stimulation output to the prevailing threshold. METHODS: Autocapture devices (Pacesetter Microny SR+/- and Regency SR+/-; Pacesetter, Solna, Sweden) and steroid-eluting epicardial pacing leads (Medtronic CapSure Epi 10366; Medtronic, Inc, Minneapolis, MN) were implanted in 14 children. Thresholds, telemetry data, evoked response, and polarization signals were obtained at discharge and follow-up, and battery service life was calculated. RESULTS: During a median follow-up of 6.5 months, autocapture pacing was applied in 12 of 14 children. The automatically adjusted pulse amplitude of autocapture devices demonstrated low-energy pacing with no significant changes between discharge and 6 months follow-up (1.1 +/- 0.3 versus 0.9 +/- 0.3 V). Autocapture-programmed pacemakers had calculated life spans of 7.8 +/- 1.4 years (Microny) and 21.0 +/- 1.6 years (Regency). No adverse effects were noted. CONCLUSIONS: Autocapture-controlled pacing with bipolar epicardial pacing leads is feasible and safe in children. Autocapture programming results in substantial energy savings and extends battery life markedly.

Successful thrombolysis of an occluded modified Blalock shunt three days after operation.

A 10-day-old boy with pulmonary atresia received a right-sided aortopulmonary polytetrafluoroethylene shunt. Three days after the operation he became cyanotic and was reintubated. Shunt occlusion was confirmed with angiography. Recombinant tissue plasminogen activator was given locally into the proximal end of the shunt. The thrombus was completely resolved after 2 days. When administration of recombinant tissue plasminogen activator was stopped, heparin infusion was started for 5 days. Shunt patency was demonstrated by angiography at 3 months postoperatively.

Purity of recycled fibre-based materials.

In order to study the purity of recycled fibre-based materials, products containing recycled fibre as well as recycled pulp were examined with regard to their chemical impurities, toxicity and microbiological quality. The study was carried out to clarify both qualitatively and quantitatively the variations in microbiological quality. The levels of several classes of chemical substances were analysed and semi-volatile and volatile substances identified. The toxicity and mutagenicity of virgin fibre and recycled fibre materials were screened using the Photobacter toxicity test and the Ames Salmonella mutagenicity test. Preliminary chemical characterization of the mutagens was carried out. Identification of the compounds found in the mutagenic fractions was performed by gas chromatography/mass spectrometry (GC/MS). The concentration of various substances analysed was found to be low, although the variety of substances present appeared to be very broad. Preliminary chemical characterization revealed that some samples contained compounds known to have mutagenic or other toxic activity. Also, the recycled fibre pulps contained large amounts of various microbes, the microbial load consisting mainly of aerobic spore-forming bacteria. The paper-making process was found to clearly have reduced the total microbial counts.

Management of the univentricular connection: are we improving?

OBJECTIVE: To assess the impact of the bidirectional cavopulmonary shunt, total cavopulmonary connection, and baffle fenestration on outcome of the Fontan operation in our unit. METHOD: We reviewed 123 bidirectional cavopulmonary shunts and 264 Fontan operations performed from 1980 to 1995. Analysis of pulmonary artery size (right and left main and lower lobe branches) before and after bidirectional cavopulmonary shunt was performed. Outcome of the Fontan operation was analysed in various time periods to assess the effect of prior bidirectional cavopulmonary shunt, use of the lateral tunnel, and fenestration. RESULTS: Operative risk for the bidirectional cavopulmonary shunt was 4% (CI = 2-10%) with a survival of 89% (CI = 83-95%) at 36 months. Probability of conversion to Fontan operation at 36 months was 49% (CI = 38-61%). Pulmonary artery size (Nakata and lower lobe indices) fell after bidirectional cavopulmonary shunt (P = 0.0006). Fontan risk dropped from 8.5% (1980-1987) to 1.8% (1988-1995) (P = 0.02), coinciding with the use of the bidirectional cavopulmonary shunt. There was no further risk reduction after introduction of the lateral tunnel and baffle fenestration, although these comparisons are limited by relatively small numbers. Duration of hospital stay related to pleural effusions was lowest for patients with a fenestrated lateral tunnel operation (P < 0.05). CONCLUSION: The bidirectional cavopulmonary shunt is a suboptimal stimulus for pulmonary artery enlargement, but may reduce the risk of Fontan operation in selected children. Fenestrated lateral tunnel operations have reduced the duration of postoperative pleural effusions.

A biodegradable patch used as a pericardial substitute after cardiac surgery: 6- and 24-month evaluation with CT.

Biodegradable patches made of polyhydroxybutyrate (PHB) were recently shown to limit postoperative pericardial adhesions when inserted into the pericardium in sheep. The present study was carried out to evaluate the presence of post-operative retrosternal adhesions after cardiac surgery in patients operated on with the PHB patch compared with those operated on without the insertion of such a patch. 50 patients admitted for CABG and/or valvular replacement were randomised to treatment either with pericardial closure with a patch or with the pericardium left open. In 39 of these (19 with and 20 without a PHB patch) computerised tomography was carried out six and twenty-four months postoperatively. Computerised tomography indicated a lower incidence (p < 0.05) of postoperative adhesions between the patch and the cardiac surface in the patch group, compared with the corresponding area in the non-patch group. A reduction of patch size in 27% (p = 0.003) was also seen.

Nickel release from metals, and a case of allergic contact dermatitis from stainless steel.

The prevalence of allergic contact dermatitis (ACD) caused by nickel is increasing. The probable cause is the increased use of nickel-containing metals in intimate contact with the skin. The critical factor is the amount of nickel released from these metals (bioavailable nickel) onto the skin. In the present study, we determined, with flame atomic absorbtion spectrometry, the amount of nickel released into synthetic sweat from metal samples. The results of this method were compared with the results of the dimethylglyoxime (DMG) test, which is considered to be a reliable means of identifying whether nickel-containing metals may cause allergy symptoms in sensitive individuals. Out of 10 samples studied, only small amounts (< 0.5 microgram/cm2/week) were released from 2 samples, and the DMG test was negative. From 5 samples, more than 0.5 microgram/cm2/week was released, and the DMG test was positive. For 3 samples, however, the DMG test was negative, though the flame atomic absorption spectrometry test showed considerable release of nickel. Therefore, although the DMG test can be used as a first line test for determining nickel release, some DMG-negative metal materials probably induce nickel sensitization, and should by no means be advertised as safe in this respect. We also report a nickel-allergic patient who developed ACD from stainless steel, indicating that some types of stainless steel release enough nickel to elicit allergic symptoms.

Low-molecular-weight heparin (Fragmin) versus heparin for anticoagulation during cardiopulmonary bypass in open heart surgery, using a pig model.

Fragmin and heparin were studied in pigs during 120 min of cardiopulmonary bypass (CPB) and up to 240 min postoperatively, with respect to clotting, bleeding and the effects of protamine. Thirty-three pigs received bolus injections of 300 IU/kg with or without additional dosage during CPB and with or without subsequent protamine sulphate. Doses of Fragmin 60% higher were necessary to prevent clotting. These had 100% higher anti-FXa levels but about 50% shorter activated coagulation time (ACT) compared with heparin. Anti-FXa increased with cumulative doses of heparin and Fragmin but ACT and activated partial thromboplastin time (aPTT) did not, indicating a larger loss of thrombin inhibition compared with anti-FXa in both drugs during CPB. Thrombin inhibition was crucial for prevention of clotting. Protamine efficiently normalized ACT in the Fragmin group but left a residual 20% anti-FXa, which did not increase the bleeding tendency. Fragmin could adequately be monitored with ACT and would be a safe alternative to heparin in CPB.

Enlargement of the right ventricular outflow tract and the pulmonary artery with a new biodegradable patch in transannular position.

Absorbable, nonwoven patches made from polyhydroxybutyrate (PHB) were implanted as transannular patches into the right ventricular outflow tract and pulmonary artery in 13 weanling sheep, the test group. Six sheep, in which a transannular Dacron patch was implanted, served as control group. The test and the control patches were harvested 3-24 months later. The results were documented macroscopically, by histological examination, and by scanning electron microscopy. The 6-keto-PGF1 alpha activity was measured in the regenerated tissue and compared with that in the native pulmonary artery. No aneurysms were noted. Regeneration of a neointima and a neomedia, comparable to native arterial tissue, was observed in the test group. In the control group, a neointimal layer was present but no neomedia comparable to native arterial tissue. Scanning electron microscopy disclosed differences in the endothelial structure between the test and control groups. 6-keto-PGF1 alpha activity was present in both the test and control groups. It is concluded that absorbable, nonwoven patches of PHB can be used as a scaffold for tissue regeneration in low-pressure systems. The regenerated vessel had structural and biochemical qualities in common with the native pulmonary artery.

Transatrial-transpulmonary repair of atrioventricular septal defect with right ventricular outflow tract obstruction.

Twenty patients had a repair of an atrioventricular septal defect with tetralogy of Fallot (n = 13) or double outlet right ventricle (n = 7). Mean age was 3.5 years. Surgical technique included transatrial-transpulmonary resection of right ventricular outflow tract obstruction and transatrial two patch repair of the atrioventricular septal defect. Ten patients required a transannular patch and one patient had a right ventricle-pulmonary artery conduit placed. There was no hospital mortality, and mean hospital stay was 15 days. One patient had late sudden death of unknown cause. Six patients have required reoperation because of residual ventricular septal defect (VSD), mitral incompetence, residual right ventricular outflow tract obstruction, and/or conduit stenosis. No patient was reoperated on because of left ventricular outflow tract obstruction. Fifteen patients are asymptomatic, one has exertional dyspnea, and two have intermittent occasional bronchospasm. The transatrial-transpulmonary two patch repair and extensive relief of right ventricular outflow tract obstruction have given good immediate results. Reoperation rate has been high mainly due to residual VSD and mitral incompetence.

Recent results with the modified Fontan operation.

From Jan 1989 to December 1991, 61 patients had modified Fontan procedures for a variety of lesions, including tricuspid atresia (18), mitral atresia (3), double inlet ventricle (14), isomeric hearts (4) and others (22). The hospital mortality was 1.6% (70% CL.0.2-5.3%). There were 2 late deaths. The median age at operation was 3.7 years (mean 5.6, range 1.5 to 20.3 years). There were two late failures at 2 months and 30 months after the operation, requiring take down of the Fontan in one and heart transplantation in the other. The 89% of the patients followed up are in NYHA class I or II at a mean follow up of 21.5 months (range 3-35 months). This suggests that the modified Fontan operation performed with the current patient selection criteria can be performed with a low mortality and that the early results are encouraging in the majority of the patients.

Absent pulmonary valve syndrome: operation in infants with airway obstruction.

From 1979 through 1991, 19 infants with absent pulmonary valve syndrome and airway obstruction were seen for surgical treatment. All patients underwent extensive pulmonary artery aneurysmorrhaphy using cardiopulmonary bypass. Fourteen patients had simultaneous transatrial ventricular septal defect (VSD) closure, infundibular resection, and placement of a short transannular patch; 2 had transventricular VSD closure and infundibular resection without a transannular patch; 1 underwent transventricular VSD closure and transannular patching; and 2 underwent pulmonary artery aneurysmorrhaphy alone with the VSD left open. All 19 infants had good hemodynamics when taken from the operating theater, but 3 died postoperatively of severe airway obstruction, despite further tracheobronchopexy procedures in 2 (hospital mortality rate, 16%; confidence limits, 7% to 29%). Among the 16 patients discharged from the hospital, there was one late death. Five other patients have required reoperation for branch pulmonary artery stenosis (n = 2), residual airway obstruction resulting from persistent pulmonary artery dilatation (n = 1), closure of VSD (n = 1), and homograft valve insertion for pulmonary incompetence and right ventricular dysfunction (n = 1). There are 15 long-term survivors. Eight of them have episodic bronchospasm of mild to moderate severity, and all are responsive to sympathomimetic bronchodilator aerosols. The remaining 7 are asymptomatic.

Prevention of postoperative pericardial adhesions by closure of the pericardium with absorbable polymer patches. An experimental study.

Pericardial adhesions after cardiac operations are a widely known phenomenon. They may severely complicate reoperations, making reentry hazardous, increasing bleeding, and prolonging the operation time. The anatomic orientation and visibility of both bypass grafts and coronary arteries are also impaired. With the aim of minimizing pericardial adhesions after cardiac operations, we studied the course of tissue regeneration after implantation of a new absorbable patch made from poly-hydroxy-butyrate. A total of 23 sheep were studied. Of these, 18 formed the test group and five served as control animals. The animals were killed at intervals of 2 to 30 months after the operation. In 14 of the 18 test animals no adhesions developed. In three animals loose adhesions were found, and in one with signs of postoperative infection there were moderate, generalized adhesions. All control sheep showed moderate adhesions; no infection was noted in this group. Light microscopy in the test group revealed a layer of mesothelium-like cells facing the epicardial side; this was already present in the early specimens. Poly-hydroxy-butyrate appeared to be slowly phagocytosed by polynucleated macrophages, which were still found occasionally in the late samples. Lymphocytes and platelets were rare. Scanning electron microscopy showed, on the epicardial side of the regenerated tissue, a mesothelium-like lining that completely covered the underlying collagen layer. The surface cell morphology grossly resembled that of native pericardium. It was concluded that in this animal model poly-hydroxy-butyrate pericardial patches decreased adhesions and preserved coronary anatomy. The findings in the control group demonstrated that pericardial surgery in the sheep was associated with adhesion formation.

Regeneration of pericardial tissue on absorbable polymer patches implanted into the pericardial sac. An immunohistochemical, ultrastructural and biochemical study in the sheep.

A new absorbable polymer prepared from polyhydroxybutyrate (PHB) was inserted as a pericardial patch in sheep to serve as a temporary scaffold for regeneration of pericardial tissue. Postoperative adhesions were rare or absent. The present study focuses on characterization of the regenerated surface cells. The luminal surface of the regenerated tissue was covered with a complete layer of mesothelium-like cells which at light and scanning electron microscopy resembled those in native pericardium. Immunohistochemical stainings for cytokeratin and thrombomodulin were positive in these cells. Heparan sulfate proteoglycan was found in a basement-membrane-like structure beneath the surface cells, as in the normal pericardium. Transmission electron microscopy of the regenerated surface revealed cells with the characteristics of mesothelium. Prostacyclin production in the regenerated tissue was similar to that in native pericardium. The results indicate regeneration of a mesothelial layer with many of the important functions of native mesothelial cells. This may explain the presently and previously observed prevention of pericardial adhesions after cardiac surgery in this field. Clinical testing of PHB patches as pericardial substitutes is warranted in cardiac surgery when pericardial closure is desired.

A new biodegradable patch for closure of atrial septal defect. An experimental study.

Biodegradable patches made from polyhydroxybutyrate (PHB) were used to close experimentally induced atrial septal defect in six calves. The implants were evaluated after 3-12 months macroscopically and by light and scanning electron microscopy with regard to regeneration of tissue and degradation of the polymer. At implant degradation, complete endothelial layers facing the right and left atrium were observed, with a subendothelial layer of collagen and some smooth-muscle cells. The patch was degraded by polynucleated macrophages, and 12 months postoperatively no polymer material was identifiable at ordinary light microscopy, but in polarized light small particles of polymer with persisting foreign body reaction were still seen. Scanning electron microscopy revealed a complete layer of surface cells with morphologic signs of endothelium. There was no shunt or sign of infection. Biodegradable PHB patches implanted in atrial septal defects in this experimental model thus prompted formation of regenerated tissue that macroscopically and microscopically resembled native atrial septal wall. The total degradation time exceeded 12 months.