New avenues for residual stress analysis in ultrathin atomic layer deposited free-standing membranes through release of micro-cantilevers.

The fabrication of thinnest, yet undeformed membrane structures with nanometer resolution is a prerequisite for a variety of Microelectromechanical systems (MEMS). However, functionally relevant thin films are susceptible to growth-generated stress. To tune the performance and reach large aspect ratios, knowledge of the intrinsic material properties is indispensable. Here, we present a new method for stress evaluation through releasing defined micro-cantilever segments by focused ion beam (FIB) milling from a predefined free-standing membrane structure. Thereby, the cantilever segment is allowed to equilibrate to a stress-released state through measurable strain in the form of a resulting radius of curvature. This radius can be back-calculated to the residual stress state. The method was tested on a 20 nm and 50 nm thick tunnel-like ALD Image 1 membrane structure, revealing a significant amount of residual stress with 866 MPa and 6104 MPa, respectively. Complementary finite element analysis to estimate the stress distribution in the structure showed a 97% and 90% agreement in out-of-plane deflection for the 20 nm and 50 nm membranes, respectively. This work reveals the possibilities of releasing entire membrane segments from thin film membranes with a significant amount of residual stress and to use the resulting bending behavior for evaluating stress and strain by measuring their deformation.

Improving stress corrosion cracking behavior of AZ31 alloy with conformal thin titania and zirconia coatings for biomedical applications.

Magnesium and its alloys have been widely studied as materials for temporary implant devices. However, corrosion-assisted cracking phenomena such as stress corrosion cracking (SCC) continue to prevent their mainstream use. For the first time, we explore the SCC susceptibility of Atomic Layer Deposition (ALD) coated AZ31 alloys in Simulated Body Fluid (SBF). Conformal 100 nm coatings of titania and zirconia were deposited on standard dogbone specimens and subjected to slow strain rate tests at 3.5 10-6 s-1 and a temperature of 37 °C. Remarkably, the SCC susceptibility index IUTS was reduced by 6% and 40% and the Iε was reduced by more than 70% and 76% with a titania and zirconia coating, respectively. Potentiodynamic polarization, hydrogen evolution and fracture behavior of the samples revealed the drastic corrosion reduction to be the main reason for the susceptibility reduction. We discuss the observed SCC behavior of our samples in light of the coatings' electrochemical activities, wettabilities, surface integrities and mechanical properties. This straightforward conformal surface treatment can be useful as a workaround for one of the major bottlenecks of biomedical Mg based implants and hence provides a possible pathway for making them more commonplace in the field.

Enhancement of stress corrosion cracking of AZ31 magnesium alloy in simulated body fluid thanks to cryogenic machining.

Magnesium and its alloys have recently attracted great attention as potential materials for the manufacture of biodegradable implants. Unfortunately, their inadequate resistance to the simultaneous action of corrosion and mechanical stresses in the human body have hampered their use as implant materials. This work aims at evaluating the Stress Corrosion Cracking (SCC) susceptibility of the AZ31 Mg alloy after being machined under cryogenic cooling. The SCC behaviour was evaluated by means of Slow Strain Rate Tests (SSRTs) in Simulated Body Fluid (SBF) at 37 °C. Prior to testing, a full characterization of the machined surface integrity, including microstructural observations, residual stress, nano-hardness measurements and surface texture analysis was carried out together with the assessment of the corrosion properties through potentiodynamic polarization curves. In addition, the morphology of the fracture surfaces after SSRTs was analysed by means of 3D optical profiler and Scanning Electron Microscopy (SEM). The improved corrosion resistance due to the increased extension of the nano-surface layer and to the compressive residual stresses represents the reason of the reduced SCC susceptibility of cryogenically machined AZ31 samples as compared to dry machined ones.

Cognitive effects of hospital calls in anaesthesiologists.

BACKGROUND: The work hours of Norwegian physicians are under scrutiny because of an increased public focus on patient safety. Ample international research indicate harmful effects of doctor fatigue based on studies on physicians working long weeks and on-call shifts of more than 30 consecutive hours. There is a lack of research on effects relevant for short or intermediate length of work weeks and call shifts. This study intended to study cognitive effects of short or intermediate duration in-hospital calls. METHODS: Eighteen anaesthesiology residents working on-call at an operation ward or an intensive care unit at Haukeland University Hospital were invited to participate. Schedules were adapted to allow for two additional experimental shifts. Participants were subjected to Cambridge Neuropsychological Test Automated Battery cognitive testing in a rested state and on three occasions after call. Amount of sleep and self-assessed sleepiness were recorded. RESULTS: Ten residents completed all four tests during 10 months. Reaction time was longer post-call. It was significantly increased only after the 18 h night call, by 21.1 and 20.5 ms for simple and five-choice reaction time, respectively. Executive function was not significantly altered post-call. Visual memory was improved post-call. Karolinska Sleepiness Score was increased by 3.3 (long day), 2.1 (short night) and 2.5 (long night) points post-call. CONCLUSION: Reaction times were increased after 18 h night calls and non-significant increases in reaction times were apparent after the other on-call shifts. Self reported sleepiness was increased post-call. We were not able to conclude whether executive function or memory was negatively affected post-call.

Cognitive impairments after critical illness.

BACKGROUND: Cognitive impairments are common after critical illness. Aetiology and effects of cognitive impairments in this setting are not fully revealed. The aim of this study was to investigate the effect of critical illness and intensive care unit (ICU) treatment on cerebral function. METHODS: Adult ICU patients with no previous history of cerebral disorders were included. Non-delirious patients scoring ≥ 24 on mini-mental state examination on ICU discharge were explored neuropsychologically using the Cambridge Neuropsychological Test Automated Battery (CANTAB) to classify cognitive impairments. Tests were repeated at 3 and 12 months. Results were compared with a normal reference population and a surgical comparison group. RESULTS: We included 55 patients. Eighteen of 28 patients were cognitively impaired, and it was not possible to classify 27 patients. The ICU survivors tested with CANTAB scored significantly lower than the reference population. They also scored worse than a surgical comparison group but significantly on only one of 10 measures. At 3 months follow-up, included patients scored significantly worse on one of 10 reported CANTAB measures. There were no differences at 12 months. We found no associations between age, co-morbidity, Simplified Acute Physiology Score II, Sequential Organ Failure Assessment score, presence of cardiovascular disease, duration of ventilatory support and length of ICU stay, and cognitive impairments. Having a cognitive impairment did not affect other outcome measures such as mortality, health-related quality of life, and institutionalization. CONCLUSIONS: Cognitive impairments are common after critical illness and may be caused by the critical illness in itself. Incidences are high after ICU discharge (64%) but drops rapidly during the first 3 months after discharge.

Cognitive dysfunction and health-related quality of life after a cardiac arrest and therapeutic hypothermia.

BACKGROUND: Evidence-based treatment protocols including therapeutic hypothermia have increased hospital survival to over 50% in unconscious out-of-hospital cardiac arrest survivors. In this study we estimated the incidence of cognitive dysfunctions in a group of cardiac arrest survivors with a high functional outcome treated with therapeutic hypothermia. Secondarily, we assessed the cardiac arrest group's level of cognitive performance in each tested cognitive domain and investigated the relationship between cognitive function and age, time since cardiac arrest and health-related quality of life (HRQOL). METHODS: We included 26 patients 13-28 months after a cardiac arrest. All patients were scored using the Cerebral Performance Category scale (CPC) and Mini-Mental State Examination (MMSE). Twenty-five of the patients were tested for cognitive function using the Cambridge Neuropsychological Test Automated Battery (CANTAB). These patients were tested using four cognitive tests: Motor Screening Test, Delayed Matching to Sample, Stockings of Cambridge and Paired Associate Learning from CANTAB. All patients filled in the Short Form-36 for the assessment of HRQOL. RESULTS: Thirteen of 25 (52%) patients were classified as having a cognitive dysfunction. Compared with the reference population, there was no difference in the performance in motor function and delayed memory but there were significant differences in executive function and episodic memory. We found no associations between cognitive function and age, time since cardiac arrest or HRQOL. CONCLUSION: Half of the patients had a cognitive dysfunction with reduced performance on executive function and episodic memory, indicating frontal and temporal lobe affection, respectively. Reduced performance did not affect HRQOL.

Gene-Gun-Mediated Transfer of Reporter Genes to Somatic Zebrafish (Danio rerio) Tissues.

We describe the use of gene-gun-mediated transfer of luciferase and green fluorescent protein (GFP) reporter genes in zebrafish (Danio rerio). Optimization of DNA transfer parameters indicated highest overall luciferase expression in epidermis and dermis using 1-µm microcarriers and 1 µg of pCMVL plasmid DNA at a delivery pressure of 200 psi. Time course studies revealed luciferase activity peaking at 18 hours and decreasing to 30% of the maximum at day 8 after DNA transfer. Onset of reporter gene (GFP) expression was detected at 13 minutes after DNA delivery, and by 65 minutes approximately 100% of the cells in the target area exhibited GFP expression. No germline association or integration events were detected in a screen of approximately 250,000 zebrafish sperm cells by fluorescence in situ hybridization at 15 or 30 days after delivery of 1 µg of pCMVL DNA, suggesting incidental male germline integration should not be considered as a risk factor when using the biolistic DNA delivery parameters and target tissues described.

Sequences from myeloblastosis-associated virus MAV-2(O) and UR2AV involved in the formation of plaques and the induction of osteopetrosis, anemia, and ataxia.

Recombinant viruses were made between myeloblastosis-associated virus MAV-2(O) and UR2AV to examine the relationship between regions of the MAV-2(O) genome and disease induction. The env-long terminal repeat (LTR) portion of MAV-2(O), when substituted into UR2AV, was sufficient to induce osteopetrosis identical to that caused by the parent MAV-2(O). When this region was reduced to the gp37 and LTR of MAV-2(O), osteopetrosis more severe than that caused by the parent virus was induced. Recombinant viruses that contained all or part of the MAV-2(O) env gene in the absence of the MAV-2(O) LTR induced a severe, chronic anemia and late-onset osteopetrosis, leading to the conclusion that the MAV-2(O) LTR, in addition to env, was required for rapid induction of osteopetrosis. A viral recombinant, pEU, which contained the gp85 segment of UR2AV substituted into MAV-2(O), induced an ataxia/cerebellar dysfunction not seen during infection with the other chimeric or parent viruses. In vitro studies of the parent and recombinant viruses demonstrated that the ability to form plaques on chicken embryo fibroblasts correlated with the presence of the MAV-2(O) gp37 and LTR except for construct pEU. When the viruses were inoculated into 10-day-old chickens, chimeras containing the env-LTR of gp37-LTR region of MAV-2(O) induced severe regenerative anemia similar to that induced by MAV-2(O). pEU was the exception, suggesting that the unique configuration of this chimera is responsible for its unusual pathogenic properties.

Assessment of the influence of NaF or 1,25-(OH)2 vitamin D3 on the proliferative bone effect of an avian osteopetrosis virus, MAV-2(O).

1,25-(OH)2 vitamin D3 (D3) and sodium fluoride (NaF) were given to chicken embryos and newly hatched chickens infected with a slow onset strain of avian osteopetrosis-inducing virus [MAV-2(O)] to determine if either agent influenced MAV-2(O)-induced proliferation of bone. Embryos were administered MAV-2(O) and treated with: 1) up to 240 micrograms NaF or up to 100 ng D3 as embryos; 2) up to 1.8 g NaF/kg or up to 9.5 micrograms D3/kg after hatching: or 3) 240 micrograms NaF as embryos and up to 1.8 g NaF/kg after hatching. Administration of MAV-2(O) alone resulted in expansion of the cortical diameter of bone. Coadministration of NaF or D3 with MAV-2(O) did not influence the change in cortical diameter seen with MAV-2(O) alone at 18 days of incubation, and 3 and 6 weeks after hatching. Increased osteoid relative to bone (hyperosteoidosis), with NaF and MAV-2(O) compared to MAV-2(O) alone, and NaF compared to untreated controls reflected delayed mineralization of osteoid, a known fluoride effect. We conclude that the administration of NaF or D3 did not influence the incidence, severity or time of onset of the MAV-2(O)-induced proliferative changes of bone.

Characterization of osteosarcoma cells from two sibling large-breed dogs.

Neoplastic cells were isolated from 2 sibling Great Dane/Labrador Retriever mixed-breed dogs in which telangiectatic type osteosarcomas arose concurrently. Cells from various sites in the same osteosarcoma appeared similar in culture, but there were differences between the 2 osteosarcomas in growth characteristics and appearance of cells. Cells from 1 osteosarcoma had a small, but significant (P less than 0.05), cyclic adenosine monophosphate response to parathyroid hormone stimulation, indicating a low order of osteoblastic differentiation. Cells from the other osteosarcoma had no response to parathyroid hormone stimulation. Cells from both osteosarcomas and a concentrated cell-free filtrate from the osteosarcoma with osteoblastic differentiation were injected into nude mice, but osteosarcomas were not induced. Results of ultrastructural examination of osteosarcoma samples for viral particles were negative and supernatant fluids from cultured cells were considered negative for viral reverse transcriptase activity.

Lethality and bone alterations in chicken embryos and newly hatched chickens given bone-active agents.

Studies were undertaken to assess the chicken embryo and newly hatched chicken as models for studying the effects of bone-active agents. Initially, 1,25-dihydroxycholecaliferol (1,25[OH]2D3), sodium fluoride (NaF), parathyroid extract, epidermal growth factor, and prostaglandin E2, were tested for lethality over a broad dose range. One or 3 injections of 1,25(OH)2D3 into the yolk sac of chicken embryos resulted in death of embryos given greater than or equal to 0.1 ng/injection, whereas 0.01 ng was tolerated by the embryos. Administering 1,25(OH)2D3 intraperitoneally to newly hatched chickens as a single injection or weekly for 3 weeks resulted in no deaths at doses up to 50 ng. One or 3 IV injections of 800 micrograms of NaF were lethal to embryos, whereas injections of less than or equal to 400 micrograms were tolerated by the embryo. Giving chickens feed and water containing 2.4 g of NaF/kg was lethal, but no deaths occurred when chickens were given feed containing less than or equal to 1.2 g of NaF/kg. Mortality associated with the administration of epidermal growth factor to embryos was inconsistent, in that death occurred in embryos given a single injection of greater than or equal to 250 ng, but no deaths occurred in embryos given 3 injections at similar doses. Parathyroid extract and prostaglandin E2 were not lethal when administered to embryos and chickens in a single-injection or multiple-injection regimen. Overall, lethality in chicken embryos given a particular agent reflected the dose of bone-active agent injected, rather than the number of injections. Three of the bone-active agents were selected to characterize their microscopic bone effects in chicken embryos and chickens.(ABSTRACT TRUNCATED AT 250 WORDS)

Drug therapy against a transplantable guinea pig leukemia.

The effects of six clinically active drugs were tested against a ttansplantable leukemia in inbred strain 2 guinea pigs. Cytoxan and 6-mercaptopurine were found to elicit a therqeutic response against this leukemia based on complete tumor regression of the established tumor as well as a substantial increase in survival time. Animals dying in the untreated control and drug-treated groups revealed typical generalized lymphoblastic leukemia. However, only Cytoxan-treated animals that had relapsed exhibited central nervous system involvement originating from the arachnoid membrane. A tow-drug combination of Cytoxan and 1-(2-chloroethyl)-3(trans-4-methylcyclohexyl)-1-nitrosourea was found not only to prevent meningeal leukemia development but also to result in "curing" all animals from their leukemia. This observation was based on a complete clinical, hematological, and histopathological "remission" period up to 176 days. The administration of 1-(2-chloroethyl)-3-(trans-4-methylcyclohexyl)-1-nitrosourea alone was observed not only to control the systemic leukemia but also to prevent central nervous system involvement. No relapses occurred after the first "remission" period was achieved in the groups of animals that received 1-(2-chloroethyl)-3-(trans-4-methylcyclohexyl)-1-nitrosourea.