Comparison of various microalgae liquid biofuel production pathways based on energetic, economic and environmental criteria.

In view of the increasing demand for bioenergy, in this study, the techno-economic viabilities for three emerging pathways to microalgal biofuel production have been evaluated. The three processes evaluated are the hydrothermal liquefaction (HTL), oil secretion and alkane secretion. These three routes differ in their lipid extraction procedure and the end-products produced. This analysis showed that these three processes showed various advantages: possibility to convert the defatted microalgae into bio-crude via HTL thus increasing the total biodiesel yield; better energetic and environmental performance for oil secretion and an even increased net energy ratio (NER) for alkane secretion. However, great technological breakthroughs are needed before planning any scale-up strategy such as continuous wet biomass processing and heat exchange optimization for the HTL pathway and effective and sustainable excretion for both secretion pathways.

An economic, sustainability, and energetic model of biodiesel production from microalgae.

A new process evaluation methodology of microalgae biodiesel has been developed. Based on four evaluation criteria, i.e. the net energy ratio (NER), biodiesel production costs, greenhouse gases (GHG) emission rate and water footprint, the model compares various technologies for each step of the process, from cultivation to oil upgrading. An innovative pathway (hybrid raceway/PBR cultivation system, belt filter press for dewatering, wet lipid extraction, oil hydrotreating and anaerobic digestion of residues) shows good results in comparison to a reference pathway (doubled NER, lower GHG emission rate and water footprint). The production costs are still unfavourable (between 1.94 and 3.35 €/L of biodiesel). The most influential parameters have been targeted through a global sensitivity analysis and classified: (i) lipid productivity, (ii) the cultivation step, and (iii) the downstream processes. The use of low-carbon energy sources is required to achieve significant reductions of the biodiesel GHG emission rate compared to petroleum diesel.

Modelling a full scale membrane bioreactor using Activated Sludge Model No.1: challenges and solutions.

A full-scale membrane bioreactor (1,600 m(3) d(-1)) was monitored for modelling purposes during the summer of 2006. A complete calibration of the ASM1 model is presented, in which the key points were the wastewater characterisation, the oxygen transfer and the biomass kinetics. Total BOD tests were not able to correctly estimate the biodegradable fraction of the wastewater. Therefore the wastewater fractionation was identified by adjusting the simulated sludge production rate to the measured value. MLVSS and MLSS were accurately predicted during both calibration and validation periods (20 and 30 days). Because the membranes were immerged in the aeration tank, the coarse bubble and fine bubble diffusion systems coexisted in the same tank. This allowed five different aeration combinations, depending whether the 2 systems were operating separately or simultaneously, and at low speed or high speed. The aeration control maintained low DO concentrations, allowing simultaneous nitrification and denitrification. This made it difficult to calibrate the oxygen transfer. The nitrogen removal kinetics were determined using maximum nitrification rate tests and an 8-hour intensive sampling campaign. Despite the challenges encountered, a calibrated set of parameters was identified for ASM1 that gave very satisfactory results for the calibration period. Matching simulated and measured data became more difficult during the validation period, mainly because the dominant aeration configuration had changed. However, the merit of this study is to be the first effort to simulate a full-scale MBR plant.

Spontaneous pseudo-aneurysm of the deep femoral artery treated by a covered stent.

PURPOSE: To present the management of a spontaneous pseudo-aneurysm of the deep femoral artery by an endovascular technique. CASE REPORT: An 82-year-old man presented with a painless pulsating mass at the level of the upper right thigh without any previous history of trauma, surgery or puncture of the femoral artery. The mass proved to be a pseudo-aneurysm of the deep femoral artery. Thrombin injection with simultaneous balloon inflation at the neck of the aneurysm did not result in a long-lasting thrombosis. Since both general and epidural anaesthesia were absolutely contra-indicated, and because of severe stenotic lesions of the femoro-popliteal axis, we chose to exclude this aneurysm under local anaesthesia with a balloon-expandable covered Jo-stent in order to maintain patency of the deep femoral artery. Twenty months postoperatively, the aneurysm is still thrombosed while the patency of both the superficial and deep femoral artery is preserved. CONCLUSIONS: This case demonstrates that an endovascular approach can be an excellent treatment for aneurysms of the deep femoral artery, thereby avoiding an open surgical procedure while preserving the patency of the deep femoral artery.

Long-term results of primary stenting for long and complex iliac artery occlusions.

OBJECTIVES: To evaluate the long-term results of recanalization with primary stenting for long and complex iliac artery occlusions. DESIGN: Retrospective non-randomized study. METHODS: Between 1996 and 2004, 38 patients underwent recanalization of an occluded iliac artery with subsequent stenting for TASC B lesions in 12 patients, TASC C in 10 and TASC D in 16. Thirty-one patients had Fontaine stage 2 B, four patients had stage 3 and one patient had stage 4. Two patients (5.4%) presented with acute ischemia and received trombolysis before recanalization. Patency results were calculated using Kaplan and Meier analysis. The mean follow-up was 26 months. RESULTS: Technical success was 97.4%. Thirty-day mortality was 2.7%. The primary patency rate was 94%, 89% and 77% at 1, 3 and 5 years respectively. Three re-occlusions (8.1%) and one restenosis (2.7%) were observed during follow-up. The secondary patency (SP) rate was 100%, 94% and 94% after 1, 2 and 3 years. Fifteen patients underwent an associated procedure. A kissing stent procedure in three patients, a contralateral PTA of an iliac stenosis in 8, a femoro-femoral bypass in 2, a femoropopliteal bypass in 1 and an femoral endarterectomy in 2. The procedure related complication rate was 5.4%. CONCLUSION: Long-term results of iliac recanalization are excellent without major complications if the procedure is technically successful. The endovascular procedure can be an alternative to an iliofemoral or aortobifemoral bypass in a high risk population.

Soft tissue aneurysmal bone cyst.

A soft tissue aneurysmal bone cyst located in the right gluteus medius of a 21-year-old man is reported. On conventional radiography, the lesion demonstrated a spherically trabeculated mass with a calcific rim. On CT scan, it showed a well-organized peripheral calcification resembling a myositis ossificans. On MRI, it presented as a multilocular, cystic lesion with fluid-fluid levels. The lesion had no solid components except for intralesional septa. Although findings on imaging and histology were identical to those described in classical aneurysmal bone cyst, diagnosis was delayed because of lack of knowledge of this entity and its resemblance to the more familiar post-traumatic heterotopic ossification (myositis ossificans).

Transfer of tilapia (Oreochromis niloticus) to a hyperosmotic environment is associated with sustained expression of prolactin receptor in intestine, gill, and kidney.

Expression of the tilapia prolactin receptor (tiPRL-R) has been characterized in the intestine of Oreochromis niloticus and the levels of both tiPRL-R transcripts and tiPRL binding sites have been further analyzed in this organ, as well as in gill and kidney, during adaptation of tilapia to a hyperosmotic environment. A single high-affinity binding site for tilapia PRL-I (tiPRL-I) was determined in full-length intestine by Scatchard analysis. A heterogeneous distribution of tiPRL-R was detected in this organ, with the posterior part always displaying a higher expression of both tiPRL-R transcript and tiPRL binding sites than the anterior and medial parts. Transfer of tilapia to brackish water (BW) led to an apparent increase in the specific binding of tiPRLs in intestine and gill even for long-term-adapted fish, whereas the high level of kidney tiPRL binding sites measured in control fish reared in fresh water was still detected in BW-adapted tilapia. There was no overall significant modification of tiPRL-R transcript levels in any organ during short-term or long-term adaptation, although a limited decrease occurred in the gill of BW-adapted fish, as shown earlier. Therefore, in O. niloticus adapted to BW, high and sustained levels of tiPRL-R were observed in the three major osmoregulatory organs, gill, kidney, and intestine.

Evidence of rainbow trout prolactin interaction with its receptor through unstable homodimerisation.

This study aims to characterise Prolactin receptor (PRLR) in rainbow trout for which no information is available despite the availability of Salmonid PRL preparations. By screening a freshwater rainbow trout intestine cDNA library with a probe corresponding to the extracellular domain (ECD) of tilapia PRLR, we have cloned a 2.5 kb insert coding for the PRLR. The mature protein of 614 amino acid residues is similar to PRLR isolated in tilapia and also the long form of mammalian PRLR. Analysis of PRLR gene expression in osmoregulatory organs revealed the presence of a unique transcript, thus confirming the involvement of this hormone in the control of osmoregulation in this fish species. By using surface plasmon resonance (SPR) technology, kinetic measurement of interaction between trout PRL and its receptor ECD was studied. This approach allowed us to demonstrate the formation of a transient, unstable homodimeric complex. This unstability could explain the inability to perform binding experiments using homologous PRL. In contrast, heterologous lactogenic ligands were able to interact through a more stable complex. Whether these characteristics of PRL-receptor interaction in rainbow trout are different to what occurs in tilapia where a homologous radioreceptor assay was developed would require further studies.

Recombinant prolactin receptor extracellular domain of rainbow trout (Oncorhynchus mykiss): subcloning, preparation, and characterization.

The cDNA of the extracellular domain of rainbow trout (Oncorhynchus mykiss) prolactin receptor (trPRLR-ECD) was cloned in the prokaryotic expression vector pMON to enable its expression in Escherichia coli after induction with nalidixic acid. The bacterially expressed trPRLR-ECD protein, contained within the refractile body pellet, was solubilized in 4.5 M urea, refolded, and purified on a Q-Sepharose column, pH 8, by stepwise elution with NaCl. The bioactive monomeric 26-kDa fraction was eluted in 0.2 M NaCl, yielding 20 mg/2.5 L of induced culture. The purified protein was over 98% homogeneous, as shown by SDS-PAGE in the presence or absence of reducing agent and by chromatography on a Superdex column. Binding experiments using [125I]ovine placental lactogen (oPL) as a ligand revealed that human growth hormone (hGH), oPL, and ovine prolactin (oPRL) were the most effective competitors, with respective IC50 values of 1.32, 2.27, and 2.70 nM. Chicken (ch) PRL did not compete at all, and homologous trPRL was much less effective, with a corresponding IC50 value of 1826 nM. Gel-filtration was used to determine the stoichiometry of trPRLR-ECD's interaction with oPL, hGH, and oPRL. Only oPL yielded a 2:1 complex, whereas hGH and oPRL formed only 1:1 complexes, with excess trPRLR-ECD being seen at the initial 2:1 trPRLR-ECD:hGH or trPRLR-ECD:oPRL ratios. No studies were performed with chPRL because of its inability to compete with [125I]oPL or with trPRL because of its low affinity toward trPRLR-ECD. The present results agree with previous findings indicating, as in mammals, that homologous PRL interacts transiently with its receptor and suggest that transient homologous PRL-induced homodimerization of the receptor is sufficient to initiate a biological signal, despite the fact that, in classical binding experiments, only low specific binding can be detected.

APH-1, a POU homeobox gene expressed in the salt gland of the crustacean Artemia franciscana.

We characterized the first POU-homeoprotein in a crustacean (designated APH-1 for Artemia POU-Homeoprotein, EMBL Y15070). The amino acid sequence of the APH-1 POU-domain is identical, except for two residues, to that of the two class III POU proteins Cf1-a (Drosophila) and POU-M1 (Bombyx mori). Southern blot analysis suggests that crustaceans have only one class III POU gene. RT-PCR and whole-mount in situ hybridization show that APH-1 mRNA is present in larvae specifically in the salt gland, an organ which is involved in osmoregulation, and disappears in the adult.

Lys13 plays a crucial role in the functional adaptation of the thermophilic triose-phosphate isomerase from Bacillus stearothermophilus to high temperatures.

The thermophilic triose-phosphate isomerases (TIMs) of Bacillus stearothermophilus (bTIM) and Thermotoga maritima (tTIM) have been found to possess a His12-Lys13 pair instead of the Asn12-Gly13 pair normally present in mesophilic TIMs. His12 in bTIM was proposed to prevent deamidation at high temperature, while the precise role of Lys13 is unknown. To investigate the role of the His12 and Lys13 pair in the enzyme's thermoadaptation, we reintroduced the "mesophilic residues" Asn and Gly into both thermophilic TIMs. Neither double mutant displayed diminished structural stability, but the bTIM double mutant showed drastically reduced catalytic activity. No similar behavior was observed with the tTIM double mutant, suggesting that the presence of the His12 and Lys13 cannot be systematically correlated to thermoadaptation in TIMs. We determined the crystal structure of the bTIM double mutant complexed with 2-phosphoglycolate to 2.4-A resolution. A molecular dynamics simulation showed that upon substitution of Lys13 to Gly an increase of the flexibility of loop 1 is observed, causing an incorrect orientation of the catalytic Lys10. This suggests that Lys13 in bTIM plays a crucial role in the functional adaptation of this enzyme to high temperature. Analysis of bTIM single mutants supports this assumption.

Traumatic basilar artery dissection in a child: need for anticoagulation?

Dissection of a cerebral blood vessel is a rare complication of acute neurotrauma with a high incidence of morbidity and mortality. We report on a case of a pediatric patient with severe neurological symptoms in whom angiography showed evidence of a basilar artery dissection. The patient was heparinized and recovered uneventfully.

Identification and modulation of a growth hormone-binding protein in rainbow trout (Oncorhynchus mykiss) plasma during seawater adaptation.

A soluble protein that specifically bound 125I-human growth hormone (hGH) was identified in rainbow trout plasma, using HPLC-gel filtration. The binding affinity of the protein for hGH was 1.2 x 10(9)M-1. 125I-rainbow trout GH (tGH) was also able to bind to the protein albeit with a lower affinity (6.6 x 10(7)M-1) than hGH. Crosslinking experiments using 125I-hGH revealed two specific bands of 150 and 130 kDa. The complex 125I-hGH-BP could be precipitated by a monoclonal anti-GH receptor antibody, suggesting a close relationship between the plasma GH-BP and the GH receptor. A fourfold increase in the hGH binding to the GH-BP was shown 48 h after transfer of the fishes from freshwater to seawater. The increase in binding was related to a high binding capacity without significant changes in binding affinity. These results suggest a potential role of this related GH-BP as an index of GH effects during seawater adaptation in salmonids.

Triose-phosphate isomerase (TIM) of the psychrophilic bacterium Vibrio marinus. Kinetic and structural properties.

The purification and characterization of triose-phosphate isomerase from the psychrophilic bacterium Vibrio marinus (vTIM) is described. Crystal structures of the vTIM-sulfate complex and the vTIM-2-phosphoglycolate complex (at a 2.7-A resolution) are also presented. The optimal growth temperature of Vibrio marinus is 15 degrees C. Stability studies show that vTIM is an unstable protein with a half-life of only 10 min at 25 degrees C. The vTIM sequence is most closely related to the sequence of Escherichia coli TIM (eTIM) (66% identity), and several unique structural features described for eTIM are also seen in vTIM, but eTIM is considerably more stable. The Td values of vTIM and eTIM, determined by calorimetric studies, are 41 and 54 degrees C, respectively. Amino acid sequence comparison reveals that vTIM has an alanine in loop 8 (at position 238), whereas all other TIM sequences known to date have a serine. The vTIM mutant A238S was produced and characterized. Compared with wild type, the catalytic efficiency of the A238S mutant is somewhat reduced, and its stability is considerably increased.

Trypsin and trypsinogen from an Antarctic fish: molecular basis of cold adaptation.

Trypsin from Antarctic fish Paranotothenia magellanica displays molecular and kinetic properties typical of enzymes produced by psychrophilic organisms. The enzyme has a high catalytic efficiency at low and moderate temperatures and is rapidly inactivated at temperatures higher than 30 degrees C. The nucleotide sequence was determined after mRNA extraction and cDNA synthesis. The cDNA encodes a pretrypsinogen which includes a seven residue activation peptide containing only three acidic residues preceeding the 222 amino-acid mature enzyme. A three-dimensional model of the enzyme was built. Structural parameters possibly involved in the adaptation to cold have been derived from comparison with the three-dimensional structure of the bovine enzyme. Among them are the lack of Tyr-151 in the substrate binding pocket, an overall decrease in the number of salt bridges and hydrophobicity and the increase in the surface hydrophilicity.

Differential effects of gonadotropin-releasing hormone, dopamine and somatostatin and their second messengers on the mRNA levels of gonadotropin II beta subunit and growth hormone in the teleost fish, tilapia.

In cultured pituitary cells of tilapia, gonadotropin-releasing hormone (GnRH; 10 nM 4-24 h), elevation of cyclic AMP (by 10 microM forskolin or 0.2 mM 3-isobutyl-1-methylxanthine: IBMX 0.5-36 h) or activation of protein kinase C (PKC; by 12.5 nM tetradecanoyl phorbol-13-acetate: TPA, 0.5-24 h) all increased gonadotropin (GtH) II beta steady state mRNA levels by three to four-fold. The involvement of PKA and PKC in the GnRH stimulatory effect on both GtH release and GtH II beta mRNA levels was corroborated by use of the PKA and PKC inhibitors, H89 and GF109203X, respectively (100 nM) which attenuated the GnRH effect. Incubation with actinomycin D (8 microM, 4-21 h) after preexposure for 24 h to either forskolin (10 microM) or TPA (12.5 nM), revealed that rates of transcript degradation were slower in forskolin-treated cells (T 1/2 = 14.1 h) than in control or TPA-treated cells (T 1/2 = 8.47 or 8.38 h), suggesting a stabilizing effect on the mRNA. Dopamine (DA; 10 microM, 4-36 h) had no apparent effect on steady state mRNA levels of GtH II beta, but reduced GtH release by as much as 75%. Steady state levels of growth hormone (GH) mRNA were not affected by exposure to GnRH (10 nM, 4-24 h), although GH release was more than doubled. Similarly, activation of PKC (by TPA 12.5 nM, 1.5-36 h), which was shown to be essential for the GnRH-stimulatory effect on GH release, did not alter levels of the GH transcript, but increased GH release by more than fivefold. DA (10 microM, 4-24 h) moderately increased GH transcript levels (160%) with similar kinetics but lower potency than direct elevation of cAMP (by 10 microM forskolin or 0.2 mM IBMX, 0.5-36 h) which increased transcript levels by more than fourfold. The involvement of PKA in the DA effect was confirmed when the PKA inhibitor H89 (100 nM, 15 min prior to DA exposure) attenuated the DA effect on GH mRNA levels. Exposure of cells to actinomycin D (8 microM, 2-16 h) after treatment with forskolin (10 microM, 24 h) led to a slower rate of transcript degradation than in control cells (T 1/2 = 6.5 h vs. T 1/2 = 4.36 h), suggesting that cAMP also elicits a stabilizing effect on GH mRNA. Somatostatin (100 nM, 0.5-36 h) had no clear effect on GH transcript levels, but reduced GH release by as much as 90%. These results suggest that activation of either cAMP-PKA or PKC pathways can, possibly by different mechanisms, stimulate mRNA levels of the GtH II beta gene, but that only the cAMP-PKA pathway stimulates GH mRNA levels. It would appear therefore that GnRH, although stimulating GH release, does not regulate GH transcription in this fish.

Traumatic type B aortic dissection.

A 37-year-old man was referred with thoracic pain after a deceleration trauma. He also had a cerebral contusion and a wrist fracture. There were no sings of hypovolemic shock. Computerized tomography (CT) of the chest and transoesophageal echocardiography (TEE) demonstrated a type B aortic dissection originating just distal to the left subclavian artery. There was a patent false lumen without rupture or distal ischaemia. Conservative treatment was given. A paralytic ileus developed and abdominal complaints persisted for several months. Angiography showed normal patency of mesenteric vessels. On follow-up, 3 years after the accident a slight aortic dilation was found on CT thorax without development of a post-dissection aneurysm. Blunt thoracic injury to the aorta usually gives rise to aortic rupture in the region of the isthmus, which can be complete or partial. In the latter case a false aneurysm may develop. An intimal tear after blunt trauma leading to type B aortic dissection rarely occurs. General principles regarding treatment of type B dissection also apply to this particular condition.

Crystal structure of recombinant triosephosphate isomerase from Bacillus stearothermophilus. An analysis of potential thermostability factors in six isomerases with known three-dimensional structures points to the importance of hydrophobic interactions.

The structure of the thermostable triosephosphate isomerase (TIM) from Bacillus stearothermophilus complexed with the competitive inhibitor 2-phosphoglycolate was determined by X-ray crystallography to a resolution of 2.8 A. The structure was solved by molecular replacement using XPLOR. Twofold averaging and solvent flattening was applied to improve the quality of the map. Active sites in both the subunits are occupied by the inhibitor and the flexible loop adopts the "closed" conformation in either subunit. The crystallographic R-factor is 17.6% with good geometry. The two subunits have an RMS deviation of 0.29 A for 248 C alpha atoms and have average temperature factors of 18.9 and 15.9 A2, respectively. In both subunits, the active site Lys 10 adopts an unusual phi, psi combination. A comparison between the six known thermophilic and mesophilic TIM structures was conducted in order to understand the higher stability of B. stearothermophilus TIM. Although the ratio Arg/(Arg+Lys) is higher in B. stearothermophilus TIM, the structure comparisons do not directly correlate this higher ratio to the better stability of the B. stearothermophilus enzyme. A higher number of prolines contributes to the higher stability of B. stearothermophilus TIM. Analysis of the known TIM sequences points out that the replacement of a structurally crucial asparagine by a histidine at the interface of monomers, thus avoiding the risk of deamidation and thereby introducing a negative charge at the interface, may be one of the factors for adaptability at higher temperatures in the TIM family. Analysis of buried cavities and the areas lining these cavities also contributes to the greater thermal stability of the B. stearothermophilus enzyme. However, the most outstanding result of the structure comparisons appears to point to the hydrophobic stabilization of dimer formation by burying the largest amount of hydrophobic surface area in B. stearothermophilus TIM compared to all five other known TIM structures.