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Introduction. Prisoners are at risk of developing vitamin D deficiency due to their lacking exposure to sunlight. So far, there are no published studies evaluating blood levels of vitamin D in relation to the health status of inmates and the quality of the Italian prison system Aim. To investigate vitamin D status and its determinants in a cohort of prisoners. Subject and Methods. One hundred and seventy-two (172) prison inmates (males, n=159, age 47± 11.3 years; females, n=13, age 43.91±12.18 years) of three penitentiaries in the province of Salerno. Vitamin D deficiency, insufficiency and sufficiency were respectively defined as a 25(OH)D level <20 ng/mL; from 20 to 30 ng/mL, >30 ng/mL. Results. In our group, Vitamin D deficiency occurs in 77.32% of the prisoners with 32.55% of the cases having severe insufficiency. Prisoners with higher BMI show lower circulating vitamin D levels (p<0.001). No significant relationship was found with the du-ration of detention (Pearson R: 0.01). Conclusion. In this cohort of inmates the vitamin D status is determined by BMI, but not by the duration of the detention. [ABSTRACT FROM AUTHOR]

Background: In this study, patients treated by thoracoscopic sympathicolysis for palmar hyperhidrosis were evaluated to determine the number and response of sweat glands to intradermal acetylcholine stimulus.Methods: A total of 30 patients were included in the study. Group A consisted of 10 patients with palmar hyperhidrosis who underwent thoracoscopic sympathicolysis in October 2005, and group B consisted of 20 patients who underwent surgery during the years 1999, 2000, and 2001. The study procedure involved applying iodine alcohol to the palm and then intradermally injecting 0.1 ml 1% acetylcholine. This activated the sweat glands, which were then photographed and counted. The study procedure was performed prospectively over different periods in group A and retrospectively in group B.Results: In group A, the mean number of glands activated 1, 3, 6, and 12 months after surgery were 41, 174.20, 522.8, and 747.2, respectively; this gradual increase was statistically significant over the first 6 months (p = 0.004) but not between months 6 and 12 (p = 0.255). The trend towards an increasing number of active glands occurred in both groups, with a mean of 1369.8 active glands in group B compared to 747.2 (p = 0.095) in group A after 12 months.Conclusion: It is well-known that Cannon's law of denervation (1939) is not applicable to the sweat glands, that is, there is no hyperactivation following intradermal acetylcholine stimulation. However, some response, which increased over the first 6 months following surgery, was observed in our study. Nevertheless, this activation is subsequently self-limiting, resulting in no gland atrophy, and reinnervation occurs without patient awareness. [ABSTRACT FROM AUTHOR]

Objective: This work aims to describe the evolution of the video-assisted Thoracoscopic Surgery (VATS) approach from a multiportal access to a biportal access for thoracic herniated disc surgery. Thoracic disc herniation remains a challenging pathology for spine surgeons. VATS of the thoracic spine was described in the 90s and represented an important technical leap by including minimally invasive options for thoracic pathology. Nowadays, VATS in thoracic surgery tends to evolve towards an even less invasive technique, from a multiportal approach to a biportal one., Methods: We describe the adoption of this approach for our spinal pathology in 3 patients. We use a two-port VATS. The largest (approximately 5 cm) with an Alexis retractor and a second port (1.5 cm) just for the camera., Results: The 3 patients started walking in less than 24 hours and none suffered any complications related to the approach. All of them reported tolerable pain at the surgical site. Changing our previous VATS system from 3 to 5 ports was relatively easy regarding the surgical technique., Conclusions: This access allows the surgeon to manipulate the instrumentation confidently and the camera does not fog up as often. Extracting a piece of rib is unnecessary and theoretically, we only manipulate 1 or at most 2 intercostal nerves, so the patient's recovery is favorable., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Biodeterioration of monumental complexes is in large part due to the proliferation of various microorganisms that attack the physical-chemical structures of support materials. Various conservation and restoration interventions use commercial biocides of synthetic origin, which exhibit some human and environmental toxicity and sometimes side effects on support materials. The main objective of this work is the assessment of new biocides obtained from endemic Mediterranean plants, to be used in the preservation of cultural heritage with the goal of contributing to the sustainable use of ecosystems and to the development of Mediterranean local communities. The biocidal potential of essential oils (EOs) and solvent extracts (SEs) (ethanol and n-hexane) obtained from four plants were evaluated: Thymus mastichina (Tm), Mentha pulegium (Mp), Foeniculum vulgare (Fv), and Lavandula viridis (Lv). Microorganisms collected at an emblematic site of Portuguese cultural heritage, the ruins of the Roman city of Conímbriga, were used to evaluate the biocidal activity of the EOs and SEs. It can be concluded that (i) SEs did not exhibit fungicidal nor bactericidal activity, except for one fungus specie; (ii) biocidal activity of EOs depends on the microorganism specie. The EOs showed a relative average biocidal activity (when compared to the commercial biocide Biotin T (1% v/v)) of 64%, 32%, 30%, and 25% for Mp, Fv, Lv, and Tm. On carbonate rocks, the application of Fv and Mp EOs up to 3 layers do not promote significant color/tonality changes in the surface of the rock. And the application of three layers of Lv and four layers of Fv, Mp, and Lv OEs only promote the occurrence of blurs or stains (variation of tonality) on rocks that presents very low porosity. It can also be noted that the EO of Mp has the broadest spectrum of activity. The results allow considering the use of Mp, Fv, Lv, and Tm EOs as valid alternatives to commercial biocides, providing a prospective application in the field of green conservation of building heritage., (© 2023. The Author(s).)

Purpose: Glucuronoyl esterases (GE, family CE15) catalyse the cleavage of ester linkages in lignin-carbohydrate complexes (LCCs), and this study demonstrate how transesterification reactions with a fungal GE from Cerrena unicolor (CuGE) can reveal the enzyme's preference for the alcohol-part of the ester-bond., Methods: This alcohol-preference relates to where the ester-LCCs are located on the lignin molecule, and has consequences for how the enzymes potentially interact with lignin. It is unknown exactly what the enzymes prefer; either the α-benzyl or the γ-benzyl position. By providing the enzyme with a donor substrate (the methyl ester of either glucuronate or 4-O-methyl-glucuronate) and either one of two acceptor molecules (benzyl alcohol or 3-phenyl-1-propanol) we demonstrate that the enzyme can perform transesterification and it serves as a method for assessing the enzyme's alcohol preferences., Conclusion: CuGE preferentially forms the γ-ester from the methyl ester of 4-O-methyl-glucuronate and 3-phenyl-1-propanol and the enzyme's substrate preferences are primarily dictated by the presence of the 4-O-methylation on the glucuronoyl donor, and secondly on the type of alcohol., (© 2023. The Author(s).)

Endo-fucoidanases, including EC 3.2.1.211 endo-α-1,3-L-fucanase and EC 3.2.1.212 endo-α-1,4-L-fucanase activities, catalyze depolymerization of fucoidans - a group of bioactive, sulfated fucosyl-polysaccharides found primarily in brown macroalgae (brown seaweeds). Quantitative assessment of endo-fucoidanase activity is critical for characterizing endo-fucoidanase kinetics and for comparing the action of different endo-fucoidanases on different types of fucoidans. However, the current state-of-the-art endo-fucoidanase assay consists of a qualitative assessment based on Carbohydrate-Polyacrylamide Gel Electrophoresis. Here, we report a new quantitative endo-fucoidanase assay based on real time spectral evolution profiling of changes in substrate and product during endo-fucoidanase action using Fourier Transform InfraRed spectroscopy (FTIR) combined with Parallel Factor Analysis (PARAFAC). The FTIR-PARAFAC assay was validated by monitoring the reaction progress of three different microbial endo-fucoidanase enzymes, FcnAΔ229, FFA2 and Fhf1Δ470, on two different fucoidan substrates. The substrates were purified from the brown macroalgae Fucus evanescens and Fucus vesiculosus, respectively. The evolution profiling showed that the strongest spectral change of the fucoidans during enzymatic depolymerization occurred in the spectral range 1220-1260 cm -1 , but the profiles differed depending on the substrate and the enzyme used. Spectral changes within 1220-1260 cm -1 are in agreement with the enzymatic depolymerization inducing signature changes in the mid-infrared absorption of sulfated fucosyls as sulfate ester bonds and C-O stretching vibrations absorb in this spectral region. Based on the data obtained, we also introduce an activity unit for endo-fucoidanases: One endo-fucoidanase Unit, U f , is the amount of enzyme able to catalyze a change in the FTIR-PARAFAC score by 0.01 during 498 s of reaction (8.3 min) on 20 g/L pure fucoidan from F. evanescens at 42 °C, pH 7.4, 100 mM NaCl and 10 mM CaCl 2 . This new quantitative endo-fucoidanase assay can pave the way for better kinetic characterizations as well as novel explorations of endo-fucoidanases., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Fucoidanases are endo-fucoidanases (also known as endo-fucanases) that catalyze hydrolysis of α-glycosidic linkages in fucoidans, a family of sulfated fucose-rich polysaccharides primarily found in the cell walls of brown seaweeds. Fucoidanases are promising tools for producing bioactive fucoidan oligosaccharides for a range of biomedical applications. High sulfation degree has been linked to high bioactivity of fucoidans. In this study, a novel fucoidanase, Fhf2, was identified in the genome of the aerobic, Gram-negative marine bacterium Formosa haliotis . Fhf2 was found to share sequence similarity to known endo-α(1,4)-fucoidanases (EC 3.2.1.212) from glycoside hydrolase family 107. A C-terminal deletion mutant Fhf2∆484, devoid of 484 amino acids at the C-terminus, with a molecular weight of approximately 46 kDa, was constructed and found to be more stable than the full-length Fhf2 protein. Fhf2∆484 showed endo-fucoidanase activity on fucoidans from different seaweed species including Fucus evanescens , Fucus vesiculosus , Sargassum mcclurei , and Sargassum polycystum . The highest activity was observed on fucoidan from F. evanescens . The Fhf2∆484 enzyme was active at 20-45°C and at pH 6-9 and had optimal activity at 37°C and pH 8. Additionally, Fhf2∆484 was found to be calcium-dependent. NMR analysis showed that Fhf2∆484 catalyzed hydrolysis of α(1,4) linkages between L-fucosyl moieties sulfated on C2 (similar to Fhf1 from Formosa haliotis ), but Fhf2∆484 in addition released oligosaccharides containing a substantial amount of 2,4-disulfated fucose residues. The data thus suggest that the Fhf2∆484 enzyme could be a valuable candidate for producing highly sulfated oligosaccharides applicable for fucoidan bioactivity investigations., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Trang, Mikkelsen, Vuillemin, Meier, Cao, Muschiol, Perna, Nguyen, Tran, Holck, Van, Khanh and Meyer.)

Background: Long-lasting heavy alcohol intake has been progressively recognized as a leading cause of nonischemic dilated cardiomyopathy, involving 10% of all people who use alcohol. It is of huge importance to identify the earliest markers of this dysfunction and it is known that the newest echocardiographic techniques such as speckle tracking may allow to do it. In this study we investigated if standard basal echocardiogram features allow to obtain predictors of asymptomatic cardiac dysfunction in alcoholics., Methods: A population of 80 consecutive asymptomatic alcoholics was enrolled. None presented history, signs or symptoms of cardiovascular disease. All of them underwent a conventional transthoracic monobidimensional and doppler echocardiography., Results: Our cohort did not present echocardiographic findings of increased left ventricular sizes, mass or relative wall thickness. Hence, a significant rate of systolic dysfunction was not found. Furthermore, statistical analysis displayed an inverse relationship between alcohol consumption and systolic pulmonary arterial pressure as well as between alcohol abuse and left atrium enlargement. This may be explained by a potential vasodilator mechanism occurring in the earliest stages of alcohol intake. On the contrary, a positive correlation with the E/A ratio was found, and this might be ascribed to state of high cardiac output determined by alcohol abuse. There were modes sex-related differences., Conclusions: This study has demonstrated that standard echocardiography may allow to predict cardiac dysfunction in asymptomatic alcoholics, and sex-related differences may be identified in this regard. These data need to be confirmed by further studies involving larger population.

Laccases (EC 1.10.3.2) are enzymes known for their ability to catalyze the oxidation of phenolic compounds using molecular oxygen as the final electron acceptor. Laccase activity is commonly determined by monitoring spectrophotometric changes (absorbance) of the product or substrate during the enzymatic reaction. Fourier Transform Infrared Spectroscopy (FTIR) is a fast and versatile technique where spectral evolution profiling, i.e. assessment of the spectral changes of both substrate and products during enzymatic conversion in real time, can be used to assess enzymatic activity when combined with multivariate data analysis. We employed FTIR to monitor enzymatic oxidation of monolignols (sinapyl, coniferyl and p-coumaryl alcohol), sinapic acid, and sinapic aldehyde by four different laccases: three fungal laccases from Trametes versicolor, Trametes villosa and Ganoderma lucidum, respectively, and one bacterial laccase from Meiothermus ruber. By coupling the FTIR measurements with Parallel Factor Analysis (PARAFAC) we established a quantitative assay for assessing laccase activity. By combining PARAFAC modelling with Principal Component Analysis we show the usefulness of this technology as a multivariate tool able to compare and distinguish different laccase reaction patterns. We also demonstrate how the FTIR approach can be used to create a reference system for laccase activity comparison based on a relatively low number of measurements. Such a reference system has potential to function as a high-throughput method for comparing reaction pattern similarities and differences between laccases and hereby identify new and interesting enzyme candidates in large sampling pools., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Laccases (EC 1.10.3.2) are enzymes known for their ability to catalyse the oxidation of phenolic compounds using molecular oxygen as the final electron acceptor. Lignin is a natural phenylpropanoids biopolymer whose degradation in nature is thought to be aided by enzymatic oxidation by laccases. Laccase activity is often measured spectrophotometrically on compounds such as syringaldazine and ABTS which poorly relate to lignin. We employed natural phenolic hydroxycinnamates having different degree of methoxylations, p-coumaric, ferulic and sinapic acid, and a lignin model OH-dilignol compound as substrates to assess enzyme kinetics by HPLC-MS on two fungal laccases Trametes versicolor laccase, Tv and Ganoderma lucidum laccase, Gl. The method allowed accurate kinetic measurements and detailed insight into the product profiles of both laccases. Both Tv and Gl laccase are active on the hydroxycinnammates and show a preference for substrate with methoxylations. Product profiles were dominated by the presence of dimeric and trimeric species already after 10 minutes of reaction and similar profiles were obtained with the two laccases. This new HPLC-MS method is highly suitable and accurate as a new method for assaying laccase activity on genuine phenolic substrates, as well as a tool for examining laccase oxidation product profiles.

Four decades have passed since Harlan and Stemler (1976) proposed the eastern Sahelian zone as the most likely center of Sorghum bicolor domestication. Recently, new data on seed impressions on Butana Group pottery, from the fourth millennium BC in the southern Atbai region of the far eastern Sahelian Belt in Africa, show evidence for cultivation activities of sorghum displaying some domestication traits. Pennisetum glaucum may have been undergoing domestication shortly thereafter in the western Sahel, as finds of fully domesticated pearl millet are present in southeastern Mali by the second half of the third millennium BC, and present in eastern Sudan by the early second millennium BC. The dispersal of the latter to India took less than 1000 years according to present data. Here, we review the middle Holocene Sudanese archaeological data for the first time, to situate the origins and spread of these two native summer rainfall cereals in what is proposed to be their eastern Sahelian Sudan gateway to the Red Sea and the Indian Ocean trade., Competing Interests: Compliance with Ethical StandardsThe authors declare that they have no conflict of interest.

Objectives: A prospective, randomized study was carried out on patients undergoing lung cancer surgery, with the aim of determining if uniportal video-assisted lobectomy has more favourable postoperative outcomes than other video-assisted thoracoscopic lobectomy techniques (Duke approach and Copenhagen approach)., Methods: Patients were randomly assigned to two groups; uniportal video-assisted lobectomy (Group A; n = 51) and other video-assisted thoracoscopic lobectomy techniques (Group B; n = 55). The primary outcome measures were: postoperative pain (analogue visual scale) and supplementary doses of analgesics (morphine, milligrams); the secondary outcome measures were: the delay in removing the paravertebral catheter and the chest drain, the duration of the postoperative hospital stay, postoperative complications and the operative or 30-day mortality. We assessed postoperative pain during the first 3 days to identify possible differences coinciding with paravertebral catheter removal and with the start of mobilization, and we evaluated the type of resection, R0/R1 (a very important factor in assessing postoperative pain). All continuous data were evaluated for normality, and analysed with the Mann-Whitney U-tests or t-tests. Categorical data were analysed by Fisher's exact test., Results: One hundred and six lobectomies were completed. Both groups were comparable with respect to different clinical parameters (age, clinical stage and comorbidity), preoperative and pathological variables. The median visual analogue pain score in the first 3 days did not show statistically significant differences (respectively, P = 0.58, P = 0.64, P = 0.85). Likewise, the median morphine use in the first 3 days did not show statistically significant differences (respectively, P = 0.72, P = 0.81, P = 0.64). There was no difference in timing to remove the paravertebral catheter (P = 0.82) and the chest drain (P = 0.65) and the duration of the postoperative hospital stay (P = 0.62). There was no difference in postoperative complications (one reoperation for bleeding in Group B, P = 0.24). There was no operative or 30-day mortality in either group., Conclusions: Uniportal video-assisted thoracoscopic lobectomy does not present better postoperative outcomes than other video-assisted thoracoscopic lobectomy techniques., (© The Author 2016. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.)

Sialylated galactooligosaccharides (GOS) represent a potential infant formula ingredient, which is believed to contribute with a combination of the beneficial properties of the prebiotic GOS as well as of sialylated human milk oligosaccharides. Sialylated GOS do not exist in natural milk, but can be produced from κ(kappa)-casein glycomacropeptide (CGMP), a sialylated side stream component from cheese-making, by sialidase-catalyzed transsialylation. Using a rationally designed mutant of the sialidase from Trypanosoma rangeli, Tr13, with enhanced transsialylation activity, six different GOS preparations with a varying degree of polymerization (DP) were effectively sialylated with molar yields of 20-30% on the CGMP sialyl in batch reactions. The rate of sialylation of the individual DPs was largely dependent on the DP distribution in each GOS preparation, and Tr13 catalysis did not discriminate against large GOS molecules, providing the novelty point that GOS molecules are sialylated independently of their size by Tr13. Using CGMP, GOS, and Tr13, the production of gram-scale quantities of sialyl-GOS was achieved in 20L volume reactions. Compared to the benchmark transsialidase from pathogenic Trypanosoma cruzi, the Tr13 was significantly more thermostable. By employing an enzymatic membrane reactor, Tr13 could be recycled and after seven consecutive 1-h reaction cycles, the biocatalytic productivity of the enzyme was increased 7-fold compared to the batch reaction. Assuming that the enzyme may be specific for α-2,3-bound sialyl moieties only, and that only 50% of sialyl linkages in CGMP are α-2,3-linked, the molar yield of sialyl-GOS on the available α-2,3-bound sialyl moieties in CGMP reached 80% in the enzymatic membrane reactor system., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Adrenocortical neoplasms are classically divided into adenomas (ACA) and carcinomas (ACC). Heterogeneous appearance and greater size are criteria to suggest malignancy, along with the urinary steroid profile (USP). The presence of regression and myelolipomatous changes in adenomas (ACA-RML) can contribute to confusion with ACC and its USP remains unknown., Objective: To evaluate the features of ACA-RML in comparison with other adrenocortical neoplasms., Methods: We selected consecutive ACA (11), ACA-RML (7) and ACC (13) cases for which USP analysis was performed before surgery and tissue was available for histological evaluation (King's College Hospital, 2005-2012). Cases were classified according to WHO and Armed Forces Institute of Pathology criteria. USPs were obtained by gas chromatography/mass spectrometry. Total excretion of individual steroids and indices (sums and ratios chosen to reflect steroid metabolic activity) were compared between ACA-RML, ACA and ACC., Results: In comparison with ACA, tumours in ACA-RML were significantly larger (8·5 ± 2·4 vs 3·5 ± 1·0, P = 0·002), presented in older patients and showed relatively higher incidence in males. Mitotic figure counts were significantly lower (0·39 ± 0·04 vs 0·93 ± 0·11 in ACA, P = 0·001) and revealed higher frequency of apoptotic cells (100% vs 9% in ACA, P = 0·001). The USP of ACA-RML showed no diagnostic features of ACC. No differences from ACA were significant, but there was a tendency towards lower dehydroepiandrosterone DHA and DHA metabolites., Conclusions: ACA-RML reveals distinctive histological features and lack of USP markers of malignancy. More cases of this rare tumour may confirm differences from ACA in steroid excretion. It is important to recognize ACA-RML because its size and heterogeneous appearance raise the possibility of ACC., (© 2014 John Wiley & Sons Ltd.)

An impaired immune response in tuberculosis patients seems to be related to weight loss that coexists with an immunoendocrine imbalance. Thus, wasting is well-recognised as a prominent feature of tuberculosis (TB), which may not be reversed even after six months of treatment. Adipokines may play a role in the immune response to M. tuberculosis, and TB may impair the expression of inflammatory adipokines, such as leptin. We aimed to study patients with pulmonary TB before and six months after treatment, by measuring plasma leptin, soluble leptin receptor and adiponectin, weight and body mass index. Nineteen patients with a diagnosis of pulmonary TB were included in the study. Blood samples were obtained before and six months after treatment, to measure plasma adipokine levels. We found an increase in plasma leptin levels after treatment (p<0.05). Even though BMI also increased, the extent was not enough to account for the changes in the leptin levels. On the other hand, plasma soluble leptin receptor and adiponectin levels did not change significantly after treatment. In conclusion, these results suggest that active TB infection may affect the expression of leptin, in addition to the wasting that may occur in these patients, and that effective TB treatment increases circulating leptin levels, probably restoring normal immunological competence.

Objective: Identify the factors of greatest impact in patients with chest trauma., Patients and Methods: prospective study of 500 patients (425 men and 75 women) with chest trauma treated between January 2006 and December 2008. The parameters assessed include the degree of trauma, the abbreviated injury scale (AIS), the injury severity score (ISS), pre-hospital intubation, duration of mechanical ventilation, stay in the intensive care unit (ICU), number of rib fractures, presence of pulmonary contusion, haemothorax and cardio-pulmonary effects., Results: The presence of polytrauma, the number of rib fractures, the presence of flail chest, pulmonary contusion, the delay in mechanical ventilation and age were shown to be effective markers of severity., Conclusions: Thoracic injuries have a number of indicators of severity. The mortality risk is associated with an ISS >25, the presence of 3 or more rib fractures with flail chest, pulmonary contusion, the development of ARDS, and with an age >55 years., (Copyright 2009 AEC. Published by Elsevier Espana. All rights reserved.)

Objective: Spontaneous pneumomediastinum is uncommon and is traditionally considered a benign, self-limited disorder that usually occurs in young adults without any apparent precipitating factor or disease. The purpose of this study is to present 47 new cases because of their different clinical behaviours., Methods: A descriptive, retrospective case series was conducted to identify adult patients with spontaneous pneumomediastinum who were diagnosed and treated in a single institution between 2000 and 2008., Results: Forty-seven patients were identified, including 33 men and 14 women with a mean age of 27.3 years (range, 16-42 years). Acute-onset chest pain was the predominant symptom at presentation (59.5%); subcutaneous emphysema was the most common sign (42.9%). The most common predisposing factor was consumption of illegal drug (cocaine): it was not a regular and long-term abuse but an occasional trend, coinciding few hours before presenting symptoms (55%). Notably, in one case, a bad outcome with mediastinal shift and important tracheal compression necessitated a right thoracotomy to open the mediastinal pleura., Conclusions: (1) It is known that occasional consumption of cocaine causes diffuse alveolar damage, haemorrhage, etc. The cause-effect relationship consumption-appearance in a few hours of spontaneous pneumomediastinum leads us to appreciate this form of consumption as a causal factor rather than a predisposing/precipitating factor as it appears in the literature on the topic. (2) Spontaneous pneumomediastinum is considered a benign disorder but, as shown by our series, a bad outcome may represent an immediate life-threatening condition, forcing an invasive manoeuvre. We always recommend performing a chest CT scan on admission to predict the evolution., (Copyright (c) 2009 European Association for Cardio-Thoracic Surgery. Published by Elsevier B.V. All rights reserved.)