Sugar production from barley straw biomass pretreated by combined alkali and enzymatic extrusion.

A pretreatment that combines a thermo-mechanical process (extrusion) with chemical and biological catalysts to produce fermentable sugars from barley straw (BS) biomass was investigated. BS was firstly extruded with alkali and then, the pretreated material (extrudate) was submitted to extrusion with hydrolytic enzymes (bioextrusion). The bioextrudate was found to have 35% (w/w dwb) of total solids in soluble form, partly coming from carbohydrate hydrolysis during bioextrusion. About 48% of soluble solids dry weight is comprised by sugars, mostly glucose and xylose. Further enzymatic hydrolysis of bioextrudate could be successfully carried out at high solid loading level of 30% (w/v), with sugar production yield of 32 g glucose and 18 g xylose/100g bioextrudate at 72 h incubation (equivalent to 96 and 52 g/l concentration, respectively). These results, together with the high level of integration of the process, indicate a great potential of this pretreatment technology for sugar production from lignocellulosic substrates.

Ethanol production from glucose and xylose obtained from steam exploded water-extracted olive tree pruning using phosphoric acid as catalyst.

In this work, the effect of phosphoric acid (1% w/w) in steam explosion pretreatment of water extracted olive tree pruning at 175°C and 195°C was evaluated. The objective is to produce ethanol from all sugars (mainly glucose and xylose) contained in the pretreated material. The water insoluble fraction obtained after pretreatment was used as substrate in a simultaneous saccharification and fermentation (SSF) process by a commercial strain of Saccharomyces cerevisiae. The liquid fraction, containing mainly xylose, was detoxified by alkali and ion-exchange resin and then fermented by the xylose fermenting yeast Scheffersomyces stipitis. Ethanol yields reached in a SSF process were close to 80% when using 15% (w/w) substrate consistency and about 70% of theoretical when using prehydrolysates detoxified by ion-exchange resins. Considering sugars recovery and ethanol yields about 160g of ethanol from kg of water extracted olive tree pruning could be obtained.

Effect of water extraction on sugars recovery from steam exploded olive tree pruning.

Biomass of olive tree pruning can be considered a suitable raw material for the production of ethanol due to its high content of potentially fermentable carbohydrates. However its high extractives content could cause condensation reactions between extractives and acid insoluble lignin during pretreatment, hindering the enzymatic hydrolysis of pretreated material. In this work, the effect of extractives removal before steam explosion of olive tree pruning was evaluated. The objectives are to recover as much glucose as possible in the extraction stage and to avoid the condensation reactions. The effect of temperature and time of water extracted material on sugars recovery was studied using a response surface method according to a central composite design. Extractive removal previous to steam explosion resulted in 20% more total sugars recovery in comparison to a material without water extraction stage.

Adaptation of the xylose fermenting yeast Saccharomyces cerevisiae F12 for improving ethanol production in different fed-batch SSF processes.

An efficient fermenting microorganism for bioethanol production from lignocellulose is highly tolerant to the inhibitors released during pretreatment and is able to ferment efficiently both glucose and xylose. In this study, directed evolution was employed to improve the xylose fermenting Saccharomyces cerevisiae F12 strain for bioethanol production at high substrate loading. Adapted and parental strains were compared with respect to xylose consumption and ethanol production. Adaptation led to an evolved strain more tolerant to the toxic compounds present in the medium. When using concentrated prehydrolysate from steam-pretreated wheat straw with high inhibitor concentration, an improvement of 65 and 20% in xylose consumption and final ethanol concentration, respectively, were achieved using the adapted strain. To address the need of high substrate loadings, fed-batch SSF experiments were performed and an ethanol concentration as high as 27.4 g/l (61% of the theoretical) was obtained with 11.25% (w/w) of water insoluble solids (WIS).

Outbreak of Legionnaires' disease associated with a supermarket mist machine.

An outbreak of Legionnaires' disease affected 12 customers of a supermarket in a town in Catalonia, Spain, between August and November 2006. An epidemiological and environmental investigation was undertaken. Preliminary investigation showed that all patients had visited the same supermarket in this town where a mist machine was found in the fish section. Water samples were collected from the machine and from the supermarket's water distribution system when high-risk samples were excluded. Environmental samples from the mist machine and clinical samples from two patients tested positive for L. pneumophila serogroup 1 and had the same molecular pattern. The PFGE pattern detected in the clinical and mist-machine isolates had never previously been identified in Catalonia prior to the outbreak and has not been identified since. Four days after turning off the machine, new cases ceased appearing. Molecular study supports the hypothesis that the mist machine from the fish section of the supermarket was the source of infection. We believe it is essential to include exposure to mist machines in any legionellosis epidemiological survey.

A community outbreak of Legionnaires' disease associated with a cooling tower in Vic and Gurb, Catalonia (Spain) in 2005.

We report the investigation of a community-acquired outbreak of Legionnaires' disease. An epidemiological, environmental, and meteorological investigation was undertaken. Fifty-five cases were reported in October and November 2005. The exposure occurred in a large area, with 12 cases (21.8%) located between 1,800 and 3,400 metres from the source. Water sample cultures showed that Legionella pneumophila serogroup 1 (Lp-1) was present in five cooling towers in two industrial locations in Gurb (plants A and B). Two Lp-1 strains were recovered from plants A and B, but only Lp-1 strains from plant A showed a pulsed-field gel electrophoresis (PFGE) profile identical to those obtained from three of the cases. Inspection of the cooling towers in plant A revealed inadequate maintenance. Weather conditions in October 2005, with mostly high temperatures and high humidity, together with the flat terrain could have been favouring factors. This study showed a community outbreak from a cooling tower as a common source in a large area. Climate and terrain could explain the dissemination of contaminated aerosols.

Effect of different cellulase dosages on cell viability and ethanol production by Kluyveromyces marxianus in SSF processes.

This study was aimed to study the effect of commercial cellulases (Celluclast 1.5 LFG) on Kluyveromyces marxianus CECT 10875 growth and ethanol production in SSF processes. Preliminary tests carried out in glucose (50 g/L) fermentation medium showed that high enzyme amounts (2.5-3.5 FPU/mL) could cause a negative effect on K. marxianus growth rate and viable cells number. However, the maximum ethanol production was not affected and about 86% of the theoretical (22 g/L) was reached in all cases independently of the enzyme dosage. In SSF experiments, cell viability was always affected by enzyme loading. Nevertheless, slight differences observed on cell viability during glucose fermentation processes with the detected concentrations of the additives did not justify the negative effect observed in SSF experiments.

Community outbreak of Legionnaires disease in Vic-Gurb, Spain in October and November 2005.

This paper reports the investigation of a community-acquired outbreak of Legionnaires' disease in the municipalities of Vic and Gurb (Central Region of Catalonia, Spain). There were 55 cases reported in October and November 2005. An epidemiological and environmental investigation was undertaken. Thirty-five case patients (64%) lived in Vic or Gurb, while 36% had visited or worked in Vic or Gurb during the 10 days before onset of symptoms, but no commonly frequented building could be identified. Water probes for culture were obtained from 30 cooling towers. In five cooling towers of two industrial settings in Gurb (plants A and B), Legionella pneumophila (Lp) serogroup 1 was present. Two Lp-1 strains were recovered from cooling towers in plants A and B. The Lp-1 strain from plant A showed a PGFE profile identical with those obtained from three patients. The exposure to Legionella pneumophila apparently occurred in a large area, since 43 of the 55 cases lived, visited or worked within a distance of 1,800 m from plant A, and six cases in a distance between 2,500 and 3,400 m. The inspections of cooling towers in plant A revealed inadequate disinfectant doses of biocide, non-existent maintenance records on weekends and wrong sample points for routine microbial check-ups. Weather conditions in October 2005 template temperature and high humidity (wind conditions are unappreciable) could have been favourable factors in this outbreak together with the flat terrain of Gurb and Vic area, explaining the extensive horizontal airborne dissemination of contaminated aerosols. The outbreak could have been prevented by proper and correct maintenance of the cooling tower at plant A.

Application of Fenton's reaction to steam explosion prehydrolysates from poplar biomass.

The application of Fenton's reaction to enhance the fermentability of prehydrolysates obtained from steam explosion pretreatment of poplar biomass was studied. Reaction conditions of temperature and H2O2 and Fe(II) concentrations were studied. The fermentability of prehydrolysate treated by Fenton's reaction was tested by using different inoculum sizes of thermotolerant strain Kluyveromyces marxianus CECT 10875. The highest percentages of toxic compound degradation (ranging from 71 to 93% removal) were obtained at the highest H2O2 concentration tested (50 mM). However, a negative effect on fermentability was observed at this H2O2 concentration at the lower inoculum loading. An increase in inoculum size to 0.6 g/L resulted in an enhanced ethanol fermentation yield of 95% relative to control.

Methylenetetrahydrofolate reductase C677T polymorphism and Factor V Leiden variant in Mexican women with preeclampsia/eclampsia.

The etiology of preeclampsia is still a matter of controversy. An association between hyperhomocysteinemia and preeclamptic patients has been described. A common missense mutation in the methylenetetrahydrofolate reductase (MTHFR) gene is associated with increased plasma homocysteine concentrations. In addition, the polymorphism of gene encoding for Factor V Leiden G1691A is associated with a prothrombotic state in heterozygous subjects. Both mutations in these thrombophilic proteins appear to have different prevalence in the general population and in patients with preeclampsia/eclampsia (PE/E). We studied single nucleotide polymorphisms for MTHFR C677T and coagulation Factor V Leiden in 33 Mexican patients with PE/E as a genetic risk factor for these diseases, comparing with a normotensive pregnant control group. The genotype and allele frequencies of MTHFR C677T and Factor V Leiden mutations between Mexican women with PE/E and healthy controls were not different. We conclude that these polymorphisms do not contribute in the etiology of PE/E as it has been reported in other populations.

Neurotransmitter amino acid in cerebrospinal fluid of patients with dementia with Lewy bodies.

The purpose of the present study was to compare neurotransmitter amino acid concentrations in the cerebrospinal fluid (CSF) and CSF/plasma ratios in 21 patients with dementia with Lewy bodies (DLB) and 26 matched controls. To this purpose, we used an ion-exchange chromatographic method. DLB patients exhibit higher CSF concentrations of asparagine (+25%) and glycine (+21%) compared to a control group, whereas no differences in CSF/plasma ratios were found between both groups. On the other hand, no alterations in concentrations of glutamate, aspartate and GABA were detected in CSF of patients compared to a control group. There was no correlation among amino acid levels and CSF/plasma ratios with age, age of onset, body mass index, duration of the disease or scores of the Mini Mental State Examination, UPDRS and Hoehn and Yahr stage. These results suggest a possible role of glycine and asparagine in the pathogenic mechanism of dementia with Lewy bodies.

Establishment and characterisation of a human carcinoma cell line with acquired resistance to Aplidin.

Aplidin (APL) is a new antitumoral drug from marine origin currently in phase II clinical trials against a wide multiplicity of cancers. As resistance may be, as with other drugs, an important obstacle to the APL therapeutic efficacy, we have established an acquired resistance cellular model by continuous exposure of HeLa cells to the drug. The stably resistant subline generated (HeLa-APL), possessing more than 1000-fold relative resistance to APL than parental cells, did not show crossresistance to a subset of clinically relevant antitumoral agents. In addition, resistance was not related to overexpression of P-glycoprotein or differences in overall drug accumulation. Comparing to parental cells, HeLa-APL cells did not present either significant differences in the growth rate or apparent alterations in the cell cycle distribution. Aplidin induced rapid and persistent phosphorylation of both JNK and p38 MAPKs, resulting in activation of the mitochondrial apoptotic pathway in parental cells, but, notably, in HeLa-APL-resistant cells MAPKs activation only occurred in a slight and transiently manner, failing to activate the above-mentioned apoptotic machinery. These results suggest that sustained activation of JNK and p38 is essential for triggering the apoptotic programme induced by APL and that HeLa-APL cells bypass this apoptotic response by preventing the specific mechanisms that prime and sustain the long-term activation of these signalling cascades. Although far from human tumour physiology in vivo, HeLa-APL cells represent a potentially useful tool in gaining insights into the mode of action of APL, in selecting non-crossresistant APL structural analogues, as well as in investigating and developing methods to prevent resistance to this drug.

Factors promoting colonization by legionellae in residential water distribution systems: an environmental case-control survey.

As part of a case-control study of community-acquired Legionnaires' disease, several factors related to residential water distribution systems and public drinking water systems were studied in the homes of 124 patients with community-acquired Legionnaire's disease and in the homes of 354 controls. The presence of water reservoirs and hot water tanks was studied in residential systems. Factors such as deficient chlorine levels, pipe repairs and other work, water flow interruptions, the use of alternative water sources, inadequate cleaning operations in public water reservoirs, and the position of the home within the public network (and whether this location constituted an endpoint) were studied in public water supply systems. Levels of legionellae in domestic water samples were also measured. Although the use of water reservoirs and hot water tanks promotes colonization by legionellae in residential systems, none of the variables studied seems to increase the incidence of community-acquired Legionnaires' disease.

Cerebrospinal fluid and plasma concentrations of nitric oxide metabolites are increased in dementia with Lewy bodies.

We compared cerebrospinal fluid (CSF) and plasma concentrations of nitric oxide metabolites (NO(-)(x)) in 22 patients with dementia with Lewy bodies (DLB) and 13 matched controls. We found a pronounced increase in NO(-)(x) CSF and plasma levels in DLB patients. No changes were found in L-arginine and L-citrulline levels in CSF or plasma. There was no correlation between CSF and plasma levels and age, age of onset, duration of the disease or scores of the MiniMental State Examination. These findings reveal that excessive nitric oxide production may be related to the pathogenesis of DLB.

Serum levels of coenzyme Q in patients with Lewy body disease.

The purpose of the present study was to compare serum levels of coenzyme Q(10) (CoQ(10)) and the coenzyme Q(10) cholesterol (CoQ(10)/cholesterol) ratio in 18 patients with Lewy body disease (LBD) with 20 matched controls. The mean serum coenzyme Q10 levels in patients with LBD were significantly reduced with respect to control group, however, no differences were found in CoQ(10)/cholesterol ratio between LBD patients and control group. There was no correlation among CoQ(10) and CoQ(10)/cholesterol ratio with age, age of onset, body mass index, duration of the disease or scores of the Mini Mental State Examination, UPDRS and Hoehn and Yahr stage. These results suggest the involvement of this enzimatic system in the pathogenic mechanism of LBD.

Are halocarboranes suitable for substitution reactions? The case for 3-I-1,2-closo-C(2)B(10)H(11): molecular orbital calculations, aryldehalogenation reactions, (11)B NMR interpretation of closo-carboranes, and molecular structures of 1-Ph-3-Br-1,2-closo-C(2)B(10)H(10) and 3-Ph-1,2-closo-C(2)B(10)H(11).

In this paper, the chemistry of 3-X-1,2-closo-C(2)B(10)H(11) (X = halogen) derivatives is extended. Molecular orbital and (11)B and (13)C NMR calculations on these species are presented. A qualitative interpretation of the (11)B NMR spectra of closo o-carborane derivatives is also provided. The synthesis of 3-X-1-R-o-carborane (X = I, Br and R = Me, Ph) derivatives is reported, and aryldehalogenation at the B3 position is reported for the first time. The molecular and crystal structures of 1-phenyl-3-bromo-1,2-dicarba-closo-dodecaborane and 3-phenyl-1,2-dicarba-closo-dodecaborane are described.

Extinction of cocaine self-administration produces a differential time-related regulation of proenkephalin gene expression in rat brain.

The purpose of this study was to examine the time course effects of extinction of cocaine self-administration behavior on proenkephalin (PENK) gene expression in caudate-putamen nucleus (ST), nucleus accumbens (Acc), olfactory tubercle (Tu), piriform cortex (Pir), ventromedial hypothalamic nucleus (VMN), and central amygdala (Ce) as measured by in situ hybridization histochemistry. Seventy-two littermate male Lewis rats were randomly assigned in triads to one of three conditions: (1) contingent intravenous self-administration of 1 mg/kg/injection of cocaine (CONT); (2) noncontingent injections of either 1 mg/kg/injection of cocaine (NONCONT); or (3) saline yoked (SALINE) to the intake of the self-administering subject. The self-administering rats were trained to self-administer cocaine under a FR5 schedule of reinforcement for a minimum of 3 weeks. After stable baseline levels of drug intake had been reached, saline was substituted for drug. Following this first extinction period, cocaine self-administration was reinstated for an additional period of 2 weeks. Immediately after cessation of the last session of cocaine self-administration (day 0) and 1-, 5-, and 10-day after the second extinction period, animal brains in each triad were removed to be processed for in situ hybridization. PENK mRNA levels were significantly higher in the cocaine groups when compared with SALINE group in the ST, Acc, Pir, and Tu regions on days 0, 1, 5, and 10 of the extinction and lower in the Ce region of CONT group when compared to NONCONT and SALINE groups on days 1, 5, and 10 of the extinction period. In the VMN nucleus, PENK mRNA content in CONT group versus NONCONT and SALINE groups was also lower, but there were statistically significant differences only on day 5. These results suggest that changes in PENK gene expression after contingent cocaine administration might be involved in cocaine withdrawal states.

Ethanol production from lignocellulosic byproducts of olive oil extraction.

The recent implementation of a new two-step centrifugation process for extracting olive oil in Spain has substantially reduced water consumption, thereby eliminating oil mill wastewater. However, a new high sugar content residue is still generated. In this work the two fractions present in the residue (olive pulp and fragmented stones) were assayed as substrate for ethanol production by the simultaneous saccharification and fermentation (SSF) process. Pretreatment of fragmented olive stones by sulfuric acid-catalyzed steam explosion was the most effective treatment for increasing enzymatic digestibility; however, a pretreatment step was not necessary to bioconvert the olive pulp into ethanol. The olive pulp and fragmented olive stones were tested by the SSF process using a fed-batch procedure. By adding the pulp three times at 24-h intervals, 76% of the theoretical SSF yield was obtained. Experiments with fed-batch pretreated olive stones provided SSF yields significantly lower than those obtained at standard SSF procedure. The preferred SSF conditions to obtain ethanol from olives stones (61% of theoretical yield) were 10% substrate and addition of cellulases at 15 filter paper units/g of substrate.

Effect of chip size on steam explosion pretreatment of softwood.

Although considerable progress has been made in technology for converting lignocellulosic biomass into ethanol, substantial opportunities still exist to reduce production costs. In biomass pretreatment, reducing milling power is a technological improvement that will substantially lower production costs for ethanol. Improving sugar yield from hemicellulose hydrolysis would also reduce ethanol production costs. Thus, it would be desirable to test innovative pretreatment conditions to improve the economics by reducing electrical power of the milling stage and by optimizing pretreatment recovery of hemicellulose, as well as to enhance cellulose hydrolysis. The objective of this study was to evaluate the effect of chip size (2-5, 5-8, and 8-12 mm) on steam-explosion pretreatment (190 and 210 degrees C, 4 and 8 min) of softwood (Pinus pinaster).

Effect of surfactants and zeolites on simultaneous saccharification and fermentation of steam-exploded poplar biomass to ethanol.

In this work, the effect of the addition of different concentrations of Tween-80 and three different zeolite-like products on enzymatic hydrolysis, ethanol fermentation, and simultaneous saccharification and fermentation (SSF) process has been investigated. The ability of these products to enhance the effectiveness of the SSF process to ethanol of steam-exploded poplar biomass using the thermotolerant strain Kluyveromyces marxianus EMS-26 has been tested. Tween-80 (0.4 g/L) increased enzymatic hydrolysis yield by 20% when compared to results obtained in hydrolysis in absence of the additive. Zeolite-like products (ZESEP-56 and ZECER-56) (2.5 g/L) improved rates of conversion and ethanol yields in the fermentation of liquid fraction recovered from steam-exploded poplar. The periods required for the completion of fermentation were approx 10 h in the presence of zeolite-like products and 24 h in the absence of additives. The probable mode of action is through lowered levels of inhibitory substances because of adsorption by the additive.