Musculoskeletal demands in microsurgery-an explorative study comparing the ergonomics of microscope and 3D exoscope.

To assess neurosurgeons' physical demands and investigate ergonomic aspects when using microsurgical visualization devices. Six neurosurgeons performed micro-surgical procedures on cadaveric specimens using the prototype of a digital 3D exoscope system (Aeos®, Aesculap, Tuttlingen, Germany) and a standard operating microscope (Pentero 900, Zeiss, Oberkochen, Germany) at two different patient positions (semisitting (SS), supine (SP)). The activities of the bilateral upper trapezius (UTM), anterior deltoid (ADM), and lumbar erector spinae (LEM) muscles were recorded using bipolar surface electromyography and neck flexion, arm abduction, and arm anteversion angles by gravimetrical posture sensors. Perceived discomfort frequency was assessed and subjects compared the two systems in terms of usability, posture, physical and mental demands, and working precision. Using the exoscope led to reduced ADM activity and increased UTM and LEM activity during SS position. The neck was extended when using the exoscope system with lower arm anteversion and abduction angles during the SS position. Subjects reported discomfort at the shoulder-neck area less frequently and lower physical demands when using the Aeos®. However, mental demands were slightly higher and two subjects reported lower working precision. The exoscope system has the potential to reduce the activity of the ADM by changing surgeons arm posture which may be accompanied by less discomfort in the shoulder-neck area. However, dependent on the applied patient position higher muscle activities could occur in the UTM and LEM.

Performance comparison of two sensors based on surface plasmon resonance in a plastic optical fiber.

In silica optical fiber Surface Plasmon Resonance (SPR)-based sensors, an increase in fiber core diameter produces a corresponding increase in the sensitivity and Signal to Noise Ratio (SNR). In Plastic Optical Fiber (POF) realized in PMMA there are different influences of design parameters on the performance, as both sensitivity and SNR are concerned. In particular, the SNR, for different refractive index values of the analyte, in a 250 µm diameter POF is greater than the one in 1,000 µm diameter POF. On the other hand, the sensitivity, for the same refractive index values of the analyte, in a 1,000 µm diameter POF is greater than the one in a 250 µm diameter POF. The results of an experimental analysis demonstrating the above behavior are reported.

Bcs1p can rescue a large and productive cytochrome bc(1) complex assembly intermediate in the inner membrane of yeast mitochondria.

The yeast cytochrome bc(1) complex, a component of the mitochondrial respiratory chain, is composed of ten distinct protein subunits. In the assembly of the bc(1) complex, some ancillary proteins, such as the chaperone Bcs1p, are actively involved. The deletion of the nuclear gene encoding this chaperone caused the arrest of the bc(1) assembly and the formation of a functionally inactive bc(1) core structure of about 500-kDa. This immature bc(1) core structure could represent, on the one hand, a true assembly intermediate or, on the other hand, a degradation product and/or an incorrect product of assembly. The experiments here reported show that the gradual expression of Bcs1p in the yeast strain lacking this protein was progressively able to rescue the bc(1) core structure leading to the formation of the functional homodimeric bc(1) complex. Following Bcs1p expression, the mature bc(1) complex was also progressively converted into two supercomplexes with the cytochrome c oxidase complex. The capability of restoring the bc(1) complex and the supercomplexes was also possessed by the mutated yeast R81C Bcsp1. Notably, in the human ortholog BCS1L, the corresponding point mutation (R45C) was instead the cause of a severe bc(1) complex deficiency. Differently from the yeast R81C Bcs1p, two other mutated Bcs1p's (K192P and F401I) were unable to recover the bc(1) core structure in yeast. This study identifies for the first time a productive assembly intermediate of the yeast bc(1) complex and gives new insights into the molecular mechanisms involved in the last steps of bc(1) assembly.

Low cost sensors based on SPR in a plastic optical fiber for biosensor implementation.

This paper reports the fabrication and testing of two configurations of optical sensor systems based on Surface Plasmon Resonance (SPR) at the interface of a liquid sample and sandwiched structures realized starting from the exposed core of a Plastic Optical Fiber (POF). The proposed geometries have proven to be suitable for measuring the refractive indexes of liquids whose refractive index falls around 1.35. Furthermore, the proposed sensing head, being low cost and relatively easy to realize, may be very attractive for biosensor implementation.

The Rieske Iron-Sulfur Protein: Import and Assembly into the Cytochrome bc(1) Complex of Yeast Mitochondria.

The Rieske iron-sulfur protein, one of the catalytic subunits of the cytochrome bc(1) complex, is involved in electron transfer at the level of the inner membrane of yeast mitochondria. The Rieske iron-sulfur protein is encoded by nuclear DNA and, after being synthesized in the cytosol, is imported into mitochondria with the help of a cleavable N-terminal presequence. The imported protein, besides incorporating the 2Fe-2S cluster, also interacts with other catalytic and non-catalytic subunits of the cytochrome bc(1) complex, thereby assembling into the mature and functional respiratory complex. In this paper, we summarize the most recent findings on the import and assembly of the Rieske iron-sulfur protein into Saccharomyces cerevisiae mitochondria, also discussing a possible role of this protein both in the dimerization of the cytochrome bc(1) complex and in the interaction of this homodimer with other complexes of the mitochondrial respiratory chain.

Biogenesis of the yeast cytochrome bc1 complex.

The mitochondrial respiratory chain is composed of four different protein complexes that cooperate in electron transfer and proton pumping across the inner mitochondrial membrane. The cytochrome bc1 complex, or complex III, is a component of the mitochondrial respiratory chain. This review will focus on the biogenesis of the bc1 complex in the mitochondria of the yeast Saccharomyces cerevisiae. In wild type yeast mitochondrial membranes the major part of the cytochrome bc1 complex was found in association with one or two copies of the cytochrome c oxidase complex. The analysis of several yeast mutant strains in which single genes or pairs of genes encoding bc1 subunits had been deleted revealed the presence of a common set of bc1 sub-complexes. These sub-complexes are represented by the central core of the bc1 complex, consisting of cytochrome b bound to subunit 7 and subunit 8, by the two core proteins associated with each other, by the Rieske protein associated with subunit 9, and by those deriving from the unexpected interaction of each of the two core proteins with cytochrome c1. Furthermore, a higher molecular mass sub-complex is that composed of cytochrome b, cytochrome c1, core protein 1 and 2, subunit 6, subunit 7 and subunit 8. The identification and characterization of all these sub-complexes may help in defining the steps and the molecular events leading to bc1 assembly in yeast mitochondria.

Identification and characterization of cytochrome bc(1) subcomplexes in mitochondria from yeast with single and double deletions of genes encoding cytochrome bc(1) subunits.

We have examined the status of the cytochrome bc(1) complex in mitochondrial membranes from yeast mutants in which genes for one or more of the cytochrome bc(1) complex subunits were deleted. When membranes from wild-type yeast were resolved by native gel electrophoresis and analyzed by immunodecoration, the cytochrome bc(1) complex was detected as a mixed population of enzymes, consisting of cytochrome bc(1) dimers, and ternary complexes of cytochrome bc(1) dimers associated with one and two copies of the cytochrome c oxidase complex. When membranes from the deletion mutants were resolved and analyzed, the cytochrome bc(1) dimer was not associated with the cytochrome c oxidase complex in many of the mutant membranes, and membranes from some of the mutants contained a common set of cytochrome bc(1) subcomplexes. When these subcomplexes were fractionated by SDS/PAGE and analyzed with subunit-specific antibodies, it was possible to recognize a subcomplex consisting of cytochrome b, subunit 7 and subunit 8 that is apparently associated with cytochrome c oxidase early in the assembly process, prior to acquisition of the remaining cytochrome bc(1) subunits. It was also possible to identify a subcomplex consisting of subunit 9 and the Rieske protein, and two subcomplexes containing cytochrome c(1) associated with core protein 1 and core protein 2, respectively. The analysis of all the cytochrome bc(1) subcomplexes with monospecific antibodies directed against Bcs1p revealed that this chaperone protein is involved in a late stage of cytochrome bc(1) complex assembly.

[How informed is patient consent to interventional cardiology procedures. A quality assurance survey by the nurses]. / Quanto è informato il consenso dei pazienti alle procedure di emodinamica? Una survey di quality assurance degli infermieri.

BACKGROUND: A new patient consent form has recently been adopted in our Institution, with a uniformly written text to be used for all medical procedures and interventions. It is accompanied by a separate information sheet, explaining both the details and the risk/benefit profile for each specific procedure/intervention. It should be given to the patient as early as possible after the procedure/intervention is planned. Testing the effectiveness of this new information policy has been included into the quality assurance goals by our nursing staff. METHODS: From mid April to mid June 2004 a questionnaire was administered to all patients who had undergone an elective cardiac interventional procedure. The timing, manner and perceived completeness of the information received by patients was investigated by 14 yes/no or multiple choice questions. A goal of <5% deviation from a 100% standard was set for all indicators. RESULTS: Two hundred and thirty-eight valid questionnaires were obtained out of 308 consecutive procedures. Seven patients (3%) refused the questionnaire. The response rate was >90% for each question. Seventy-eight patients (33%) had a history of cardiac interventional procedures. The information sheet had been received before the procedure in 93% of cases, and this had happened in the ward in 58% of cases; the procedure had been performed at least 1 hour after receipt of the information sheet in 83% of cases. Twenty-seven patients (13%) stated they had not read the information sheet, in most cases (92%) because they felt they already knew enough. Among patients who had read the information sheet, 99% deemed it could be easily understood. Difficulties in asking questions were reported by 6% of patients. When questions had been asked, the nursing staff was addressed in 42% of cases, and the answers were rated as clear in 98% of cases. The consent form was not read at all by 13% of patients, due to alleged lack of time, and was not read completely by another 15%; 98% of those who had read it, however, found it was fairly understandable. CONCLUSIONS: The effectiveness of our new patient information policy seems to approach our quality goals, and is liable to further improvement. The nursing staff of the cardiac catheterization unit is involved in the patient information process, and has full competence to study this issue.

Implementation of radial arterial access for cardiac interventions: a strong case for quality assurance protocols by the nursing staff.

BACKGROUND: Radial arterial access is becoming increasingly popular for coronary angiography and angioplasty. The technique is, however, more demanding than femoral arterial access, and hemostasis is not care-free. A quality assurance program was run by our nursing staff, with patient follow-up, to monitor radial arterial access implementation in our laboratory. METHOD: In 973 consecutive patients, both a hydrophilic sheath and an inflatable bandage for hemostasis were used. Bandage inflation volume and time were both reduced through subsequent data audit and protocol changes (A = 175 patients; B = 297; C = 501). RESULTS: An increase was achieved in the percentage of patients with neither loss of radial pulse nor hematoma of any size (A = 81.3%, B = 90.9%, C = 92.2%, P < 0.001), and no discomfort at all (A = 44.2%, B = 75.1%, C = 89.3%, P < 0.001). Follow-up was available for 965 patients (99%), and in 956, the access site could be re-inspected at least once. There were no vascular complications. Overall, the radial artery pulse was absent at latest follow-up in 0.6% of cases (95% confidence interval 0.21-1.05%). In 460 consecutive patients with complete assessment in protocol C, a palpable arterial pulse was absent in 5% of cases at about 20 h after hemostasis. Barbeau's test was positive in 26.5% of patients (95% confidence interval 22.5-30.6%). They had a significantly lower body weight, a lower systolic blood pressure at hemostasis, and a higher bandage inflation volume; a hematoma of any size and the report of any discomfort were also more frequent. Barbeau's test returned to normal in 30% of them 3-60 days later. CONCLUSION: Our nurse-led quality assurance program helped us in reducing minor vascular sequelae and improving patient comfort after radial access. Early occlusion of the radial artery as detected by pulse oxymeter is frequent, often reversible, and may be mostly related to trauma/occlusion of the artery during hemostasis.

Evidence that the assembly of the yeast cytochrome bc1 complex involves the formation of a large core structure in the inner mitochondrial membrane.

The assembly status of the cytochrome bc(1) complex has been analyzed in distinct yeast deletion strains in which genes for one or more of the bc(1) subunits were deleted. In all the yeast strains tested, a bc(1) sub-complex of approximately 500 kDa was found when the mitochondrial membranes were analyzed by blue native electrophoresis. The subsequent molecular characterization of this sub-complex, carried out in the second dimension by SDS/PAGE and immunodecoration, revealed the presence of the two catalytic subunits, cytochrome b and cytochrome c(1), associated with the noncatalytic subunits core protein 1, core protein 2, Qcr7p and Qcr8p. Together, these bc(1) subunits build up the core structure of the cytochrome bc(1) complex, which is then able to sequentially bind the remaining subunits, such as Qcr6p, Qcr9p, the Rieske iron-sulfur protein and Qcr10p. This bc(1) core structure may represent a true assembly intermediate during the maturation of the bc(1) complex; first, because of its wide distribution in distinct yeast deletion strains and, second, for its characteristics of stability, which resemble those of the intact homodimeric bc(1) complex. By contrast, the bc(1) core structure is unable to interact with the cytochrome c oxidase complex to form respiratory supercomplexes. The characterization of this novel core structure of the bc(1) complex provides a number of new elements clarifying the molecular events leading to the maturation of the yeast cytochrome bc(1) complex in the inner mitochondrial membrane.

Dietary combination of conjugated linoleic acid (CLA) and pine nut oil prevents CLA-induced fatty liver in mice.

Conjugated linoleic acid (CLA) strongly prevents fat accumulation in adipose tissue of mice, even if hepatic fat deposition and insulin resistance are concomitantly observed. This study investigated the possibility of maintaining the antiadiposity properties of CLA while preventing adverse effects such as liver steatosis and hyperinsulinemia. To this end, mice were divided into three groups and fed a standard diet (control) or a diet supplemented with 1% CLA (CLA) or a mixture of 1% CLA plus 7.5% pine nut oil (CLA + P). The combination of CLA + P preserved the CLA-mediated antiadiposity properties (70% fat reduction), preventing hepatic steatosis and a sharp increase in plasmatic insulin starting from the eighth week of CLA treatment. The assay of both fatty acid synthesis and oxidation in the CLA + P mice revealed a time-dependent biphasic behavior of the corresponding enzymatic activities. A sudden change in these metabolic events was indeed found at the eighth week. A strong correlation between the changes in key enzymes of lipid metabolism and in insulin levels apparently exists in CLA-fed mice. Furthermore, lower levels of lipids, in comparison to values found in CLA-fed mice, were observed in the liver and plasma of CLA + P-fed animals.

Conjugated linoleic acid and hepatic lipogenesis in mouse: role of the mitochondrial citrate carrier.

Conjugated linoleic acid (CLA) is able to reduce adiposity by affecting lipid metabolism. In particular, CLA administration to mice reduces body fat mass with a concomitant lipid accumulation in the liver. We investigated the effects of CLA on the activity of the mitochondrial citrate carrier (CIC), which is implicated in hepatic lipogenesis. The transport activity of the CIC, measured both in intact mitochondria and in the proteoliposomes, progressively increased with the duration of CLA feeding. An increase in the CIC activity of approximately 1.7-fold was found in 16 week CLA-treated mice with respect to control animals. A kinetic analysis showed a 1.6-fold increase in the V(max) of citrate transport but no change in the K(m) value. Western blot experiments revealed an increase of approximately 1.7-fold in the expression of CIC after CLA treatment. A strict correlation between the increase in CIC activity and the stimulation of the cytosolic lipogenic enzymes was also found. These data indicate that the CIC may play a role in the onset of hepatic steatosis in CLA-fed mice by supplying the carbon source for de novo fatty acid synthesis.

Vascular access complications after cardiac catheterisation: a nurse-led quality assurance program.

BACKGROUND: Vascular access complications may be a cause of discomfort, prolonged hospital stay, and impaired outcomes in patients undergoing cardiac catheterisation. AIMS: To assess vascular access complication in our patients with/without the use of closure devices as a first local benchmark for subsequent quality improvement. METHODS: A nurse-led single-centre prospective survey of all vascular access complications in consecutive patients submitted to cardiac catheterisation during 4 months. RESULTS: The radial and femoral access were used in 78 (14%) and 470 (83%), respectively, of 564 procedures, and a closure device was used in 136 of the latter. A haematoma (any size) was isolated and uneventful in 9.6% of cases. More severe complications (haemoglobin loss >2 g, need for blood transfusion or vascular repair) occurred in 1.2% of cases, namely: in none of the procedures with radial access, and in 0.4% and 2.4% of femoral diagnostic and interventional coronary procedures, respectively. During complicated (n=40) vs uncomplicated (n=172) transfemoral interventions, the activated coagulation time was 309+/-83 vs 271+/-71 s (p=0.004), but the use of closure devices was similar. CONCLUSION: Severe vascular access complications in our patients were fewer than in most reports, and virtually absent in radial procedures. Vigorous anticoagulation was associated with increased complications in our patients, but closure devices were not. A new policy including both the use of the radial access whenever possible, and a less aggressive anticoagulation regimen during transfemoral interventions will be tested.

Further insights into the assembly of the yeast cytochrome bc1 complex based on analysis of single and double deletion mutants lacking supernumerary subunits and cytochrome b.

The cytochrome bc1 complex of the yeast Saccharomyces cerevisiae is composed of 10 different subunits that are assembled as a symmetrical dimer in the inner mitochondrial membrane. Three of the subunits contain redox centers and participate in catalysis, whereas little is known about the function of the seven supernumerary subunits. To gain further insight into the function of the supernumerary subunits in the assembly process, we have examined the subunit composition of mitochondrial membranes isolated from yeast mutants in which the genes for supernumerary subunits and cytochrome b were deleted and from yeast mutants containing double deletions of supernumerary subunits. Deletion of any one of the genes encoding cytochrome b, subunit 7 or subunit 8 caused the loss of the other two subunits. This is consistent with the crystal structure of the cytochrome bc1 complex that shows that these three subunits comprise its core, around which the remaining subunits are assembled. Absence of the cytochrome b/subunit 7/subunit 8 core led to the loss of subunit 6, whereas cytochrome c1, iron-sulfur protein, core protein 1, core protein 2 and subunit 9 were still assembled in the membrane, although in reduced amounts. Parallel changes in the amounts of core protein 1 and core protein 2 in the mitochondrial membranes of all of the deletion mutants suggest that these can be assembled as a subcomplex in the mitochondrial membrane, independent of the presence of any other subunits. Likewise, evidence of interactions between subunit 6, subunit 9 and cytochrome c1 suggests that a subcomplex between these two supernumerary subunits and the cytochrome might exist.