Synthesis and characterization of a crosslinked chitosan derivative with a complexing agent and its adsorption studies toward metal(II) ions

A new chitosan derivative has been synthesized by crosslinking a metal complexing agent, [6,6′-piperazine-1,4-diyldimethylenebis (4-methyl-2-formyl) phenol] (L), with chitosan (CTS). The resulting material (CCTSL) was characterized by elemental (CHN), spectral (FTIR and solid-state NMR), thermal (TGA and DTA), and structural (powder XRD and SEM) analyses. Adsorption experiments (pH dependency, kinetics, and equilibrium) of CCTSL toward various metal ions such as Mn(II), Fe(II), Co(II), Cu(II), Ni(II), Cd(II), and Pb(II) were carried out at 25 °C. The results showed that the adsorption was dependent on the pH of the solution, with a maximum capacity between pHs 6.5 and 8.5. The kinetics was evaluated by applying the pseudo-first-order and pseudo-second-order equation models and the equilibrium data were analyzed by Langmuir isotherm model. The maximum adsorption capacity was 1.21 mmol g⁻¹ for Cu(II) and the order of adsorption capacities for the metal(II) ions studied was found to be Cu(II) > Ni(II) > Cd(II)  Co(II)  Mn(II)  Fe(II)  Pb(II).     

Monitoring the (photo)genotoxicity of photosensitizer drugs: direct quantitation of single-strand breaks in deoxyribonucleic acid using an oligonucleotide chip

Oligonucleotide chip-based assays can be a sample-thrifty, time-saving, routine tool for evaluation of chemical-induced DNA strand breaks. This article describes a novel approach using an oligonucleotide chip to determine photosensitizer-induced DNA single-strand breaks. Surface coverage of fluorophore-labeled oligonucleotides on silicon dioxide chip surfaces was determined on alkaline phosphatase digestion. Fluorescence maxima (at 520 nm) of the solutions were converted to molar concentrations of the fluorescein-modified oligonucleotide by interpolation from a predetermined standard linear calibration curve. The photosensitizing activity of chlorpromazine and triflupromazine toward DNA single-strand breaks was then studied at different drug doses and also as a function of photoirradiation time. Photoinduced single-strand breaks calculated using the method described here agreed with values predicted by theoretical extrapolation of the single-strand breaks obtained for plasmid DNAs from agarose gel electrophoresis, and thereby indirectly validated the chip-based assays. Under UV irradiation (93.6 kJ/m²) chlorpromazine (0.08 mM) was found to have significant photogenotoxicity. However, triflupromazine did not exhibit any (photo)genotoxicity over the concentration range studied (0.04–0.20 mM). The method developed will be useful for quantitative screening of drug genotoxicity in terms of induction of breaks in DNA.     

Longer-term responses of a floodplain-dwelling marsupial to experimental manipulation of fallen timber loads

Fallen timber is an important component of many aquatic and terrestrial ecosystems around much of the world. Its distribution and abundance has been extensively altered, especially over the past 200 years. While correlative evidence suggests that many species require fallen timber for them to occupy locations, there are few precisely controlled field experiments that show conclusively how species respond to variation in loads. We manipulated wood loads (ranging from 0 to 80 Mg ha⁻¹) and monitored the response over 6 years of a species of marsupial, the yellow-footed antechinus Antechinus flavipes. The antechinus appeared to prefer locations having fallen-timber loads 20 Mg ha⁻¹. Antechinuses avoided bare areas (i.e. 0 Mg ha⁻¹) and areas having finer forms of woody material. Changes in densities of the antechinus in excess of those occurring in the control plots were clear after 2 years following manipulation of loads, and were even more pronounced after almost 6 years. Therefore, we are confident that higher loads are favoured by the antechinuses. There was no evidence of breeding success by females in sites with 20 Mg ha⁻¹, and that the highest, consistent breeding activity was achieved at sites with the greatest fallen-timber loads (80 Mg ha⁻¹). Our results, which experimentally were designed to address responses to fallen-timber loads, were complicated by perturbations caused by managed flooding episodes of the experimental area. Flooding appears to lead to population booms of the antechinus, most probably related to irruptions of prey species such as carabid beetles and wolf spiders, which are large-bodied and are prey for the antechinus. Nevertheless, we show that there are differences in densities in different wood-load treatments even against a background of boom-and-bust dynamics associated with flooding.     

Interleukin-6 and procalcitonin in children with sepsis and septic shock

Objectives. To examine the behavior of interleukin-6 (IL-6) and procalcitonin (PCT) and verify whether they can be used to differentiate children with septic conditions. Methods. Septic children aged between 28 days and 14 years, prospectively enrolled from 01/2004 to 12/2005, were divided into sepsis (SG; n = 47) and septic shock (SSG; n = 43) groups. IL-6 and PCT were measured at admission (T0) and 12 h later (T12h). PCT results were classed as: 0.5 ng/mL = sepsis unlikely; 0.5 to <2 = sepsis possible; 2 to <10 = systemic inflammation; 10 = septic shock. Results. Ninety children were included. At T0, there was a higher frequency of SSG with higher PCT compared with SG [SSG: 30 (69.7%) > SG: 14 (29.8%); p < 0.05]. Similar results were observed at T12h. PRISM was significantly higher for SSG patients with higher PCT than SG patients. At T0, IL-6 levels were higher in SSG [SSG: 213.10 (10.85–396.70) > SG: 63.21 (0.86–409.82); p = 0.001], but not statistically different at T12h. IL-6 levels positively correlated with PRISM score in SSG patients at admission (p = 0.001; r = 0.86). Conclusion. PCT and IL-6 appear to be helpful in early assessment of pediatric sepsis, are of diagnostic value at admission, and are related to disease severity.     

Effects of fragmentation on forest structure and litter dynamics in Atlantic rainforest in Pernambuco, Brazil

This study evaluates the effects of fragmentation on the spatial and temporal dynamics of small litterfall production in Atlantic rainforest in Pernambuco State, Brazil. Litterfall was collected for 24 months at two 0.2 ha sites, located in the forest edge zone and the forest interior, within a rainforest patch of about 300 ha. Structural parameters of both forest sites were recorded. Litter was sorted into six fractions (foliage, twigs, buds/flowers, fruits/seeds, peduncles, rest), dried and weighed. The interior forest plot contained 314 live trees with a dbh 5 cm and a stand basal area of 41.8±8.7 m², whereas the forest edge contained 211 live trees and a stand basal area of 23.4±3.6 m². Total small litterfall was extraordinarily high and totalled 12.62±4.73 t ha⁻¹ yr⁻¹ in interior forest and 14.74±2.78 t ha⁻¹ yr⁻¹ in forest edge. High litterfall rates are probably due to a pronounced periodicity, edge effects alter litterfall strongly.     

Temporal evolution of soluble Fas and Fas ligand in patients with orthotopic liver transplantation

Aim: The aim of this study was to analyze the expression levels of plasma soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients with orthotopic liver transplantation (OLT) procedures routinely performed without venovenous bypass. Methods: The sFas and sFasL were analyzed in the blood of 20 consecutive patients who underwent transplantation. Blood samples were drawn from the radial artery at serial time points before, during, and after surgery. Plasma levels of sFas and sFasL were detected by Enzyme Linked-Immuno-Sorbent Assay. Plasma aspartate transaminase (AST) and alanine transaminase (ALT) were assayed by routine clinical chemistry testing. Results: Marked elevation of plasma AST and ALT were detected at the reperfusion and postoperation time points (P < 0.001), with a peak on the first postoperative day. The mean plasma concentration of sFas and sFasL remained unchanged from preoperative to anhepatic phase (T1 to T3) (P  0.268). The sFas and sFasL concentrations were significantly higher at 15 and 60 min after reperfusion compared to the preoperative value (P  0.048). Postoperatively, sFas and sFasL concentration were decreased to preoperative levels on the first postoperative day (P  0.127). Conclusion: The sFas and sFasL seem to be involved in reperfusion injury during OLT. The understanding of Fas may provide new insights into the mechanisms of ischemia/reperfusion injury during OLT.     

Plant communities affect the species–area relationship on Carex sempervirens tussocks

The species–area relationship (SAR) has been extensively studied in a wide range of plant communities, but very few studies have directly addressed how plant communities affect the SAR and what are the underlying mechanisms. Many graminoids form distinct tussocks where many other plant species grow, but no study has investigated whether the SAR holds true for the vegetation on tussocks. In four plant communities on an abandoned subalpine pasture in the Swiss National Park, we made releves on 600 tussocks of Carex sempervirens and measured tussock basal area and other tussock traits. In all four communities, species richness on C. sempervirens tussocks was strongly positively related to tussock basal area (R²0.74), while other tussock traits explained very little (R²<0.04). Slope and intercept of the SAR on C. sempervirens tussocks differed significantly among the four communities. This was because plant communities affected richness in smaller tussocks (basal diameter <10 cm) but not that in large tussocks (basal diameter10 cm). We conclude that the SAR holds true for vegetation on C. sempervirens tussocks and changes with plant communities. Changes in the SAR on C. sempervirens tussocks are very likely because smaller tussocks are less independent of the plant communities than the larger ones, regarding disturbance or nutrients.     

The significance of persistent newly developed autoantibodies in JIA patients under long-term anti-TNF treatment

Objective: To study the significance of persistent (12 months) new autoantibodies, in Juvenile Idiopathic Arthritis (JIA) patients treated with either Infliximab (INFL) or Etanercept (ET) for 2 years. Patients-methods: 26 children under INFL (n = 12) or ET (n = 14) were prospectively studied. A large panel of autoantibodies was tested using indirect immunofluorescence (ANA, anti-dsDNA, anti-ENA, SMA, LKM, AMA, PCA, anti-R1, ATA), ELISA (ANA, anti-ENA, anti-cardiolipin, ANCA), immunoblotting assay (anti-ENA: anti-Ro, anti-La, anti-Sm, anti-URNP, anti-Jo, anti-Scl70, anti-centromere, anti-ribosomal and anti-histone) and rate nephelometry (RF). Results: Apart from the positive patients for ANA (13/26) and RF (2/26) prior to anti-TNF treatment, 6/26 patients (23%) developed new autoantibodies (SMA, anti-R1, ATA) which persisted for 12–50 months. None developed antibodies to nuclear antigens. In only one case, ATA was associated with the development of Hashimoto’s thyroiditis. Conclusions: These findings indicate that in JIA patients in contrast to adult RA patients, development of new autoantibodies to various nuclear antigens is rare. Other non relevant to rheumatic diseases autoantibodies, may appear and persist for >12 months, but very rarely they may be related to clinical entities, especially in the presence of a positive family history of autoimmunity.     

Kinetics of ammonium, nitrate and phosphorus uptake by Canna indica and Schoenoplectus validus

Canna indica L. is an upright perennial rhizomatous herb, and Schoenoplectus validus (Vahl) A. Löve and D. Löve is a tall, perennial, herbaceous sedge. The nutrient uptake kinetics of C. indica and S. validus were investigated using the modified depletion method after plants were grown for 4 weeks in simulated secondary-treated wastewater. The maximum uptake rate (I_max) and Michaelis–Menten constant (K_m) were estimated by iterative curve fitting. The I_max for NH₄N (623 μmol g⁻¹ dry root weight h⁻¹) was significantly higher than that for NO₃N (338 μmol g⁻¹ dry root weight h⁻¹) in S. validus. In contrast, no difference was observed in C. indica. The I_max values for NO₃N and NH₄N were higher in S. validus than in C. indica. A significantly lower K_m was detected for NO₃N uptake in C. indica (385 μmol L⁻¹) compared to that in S. validus (1908 μmol L⁻¹). The I_max for PO₄P did not differ between the plant species. The K_m for PO₄P was significantly higher in C. indica (157 μmol L⁻¹) than in S. validus (60 μmol L⁻¹). In conclusion, we found that S. validus preferred NH₄N over NO₃N, had greater capacity for N uptake and higher affinity for PO₄P, but C. indica had greater affinity for NO₃N. Nutrient uptake capacity is likely related to habitat preference, and is influenced by the structure of roots and rhizomes.     

Further mechanistic orientation for the oxidation reaction between alkaline permanganate and poly galacturonate methyl ester. Novel spectrophotometric tracer of intrahypomanganate(V) – Intermediate

The kinetics of formation of the green manganate(VI) intermediate complex formed during the oxidation of pectin polysaccharide (poly galacturonate methyl ester) (PGME) by in alkaline solution at pH  12 have been studied. The rate law was suggested to be: rate = . The reaction was found to be base – catalyzed and fractional second – order in [PGME]. The activation parameters have been evaluated and discussed. A reaction mechanism was suggested according to the experimental data. A new blue hypomanganate(V) coordination polymer intermediate sol was spectrophotometrically detected for the first time for pectin (poly galacturonate methyl ester) oxidation reaction. A further mechanistic presentation was proposed.     

Reinforcement of carboxyl groups in the surface of Corynebacterium glutamicum biomass for effective removal of basic dyes

The biomass of Corynebacterium glutamicum was treated with poly(amic acid) to improve the biosorption of Basic Blue 3 (BB3) from aqueous solution. The grafting of poly(amic acid) onto the biomass surface increased the density of the carboxyl groups. The UV-spectrum revealed that strong acidic (pH  2) and basic conditions (pH  11) resulted in the precipitation of BB3. Therefore, pH edge experiments were conducted only within the range 3–10; these results indicated that electrostatic attraction between carboxyl groups of C. glutamicum and BB3 dye cations was favored under alkaline conditions. From the Langmuir model, poly(amic acid)-modified biomass gave a maximum uptake of 173.6 mg/g at pH 9, compared to 52.8 mg/g by the raw biomass. The biosorption kinetics was found to be fast; with equilibrium attained within 10 min. The increase in the ionic strength strongly affected the uptake of BB3 for both forms of C. glutamicum.     

Production of superoxides and nitric oxide generation in haemocytes of mussel Mytilus galloprovincialis (Lmk.) after exposure to cadmium: A possible involvement of Na/H exchanger in the induction of cadmium toxic effects

The present study investigates cadmium (Cd) ability to enhance superoxides (O₂⁻) and nitric oxide (NO) production (as nitrites) in haemocytes of mussel Mytilus galloprovincialis as well as the possible involvement of Na⁺/H⁺ exchanger (NHE) in the induction of NADPH oxidase and NO synthase activity. PMA, a well-known PKC-mediated NADPH oxidase as well as NO synthase stimulator was also used, in order to verify Cd effects on both O₂⁻ and NO generation. According to the results of the present study, micromolar concentrations of Cd (0.05, 5, 10 and 50 μM) seemed to enhance O₂⁻ and NO generation in haemocytes of mussels. Moreover, O₂⁻ and NO generation in haemocytes exposed to Cd could be enhanced by its ability to induce reactive oxygen species (ROS) but respiratory burst activation as well. Inhibition of NO synthase with 10 μM l-NAME, significantly attenuated Cd ability to enhance O₂⁻ production and diminished NO generation, thus leading to the suggestion that Cd toxic effects, started at concentration of 50 μM, could enhance NADPH oxidase and NO synthase stimulation in haemocytes of mussels. NHE seems to play a regulatory role in the induction of either O₂⁻ or NO generation in haemocytes exposed to the metal, since its inhibition with the use of 10 μM EIPA significantly decrease both O₂⁻ and NO production. The involvement of NHE in the induction of O₂⁻ and NO generation, probably via PKC-mediated NADPH oxidase and NO synthase activation, is likely to be crucial to haemocytes exposed to heavy metals, such as Cd.     

In vivo real-time measurement of superoxide anion radical with a novel electrochemical sensor

The dynamics of superoxide anion (O₂⁻) in vivo remain to be clarified because no appropriate method exists to directly and continuously monitor and evaluate O₂⁻ in vivo. Here, we establish an in vivo method using a novel electrochemical O₂⁻ sensor. O₂⁻ generated is measured as a current and evaluated as a quantified partial value of electricity (Q_part), which is calculated by integration of the difference between the baseline and the actual reacted current. The accuracy and efficacy of this method were confirmed by dose-dependent O₂⁻ generation in xanthine–xanthine oxidase in vitro in phosphate-buffered saline and human blood. It was then applied to endotoxemic rats in vivo. O₂⁻ current began to increase 1 h after lipopolysaccharide, and Q_part increased significantly for 6 h in endotoxemic rats, in comparison to sham-treated rats. These values were attenuated by superoxide dismutase. The generation and attenuation of O₂⁻ were indirectly confirmed by plasma lipid peroxidation with malondialdehyde, endothelial injury with soluble intercellular adhesion molecule-1, and microcirculatory dysfunction. This is a novel method for measuring O₂⁻ in vivo and could be used to monitor and treat the pathophysiology caused by excessive O₂⁻ generation in animals and humans.     

Western equatorial Pacific planktic foraminiferal fluxes and assemblages during a La Niña year (1999)

A correlation between foraminiferal community dynamics and environmental conditions may provide a basis for establishing paleoclimatic proxies. We studied planktic foraminiferal shell fluxes and assemblages in samples collected in three time-series sediment trap deployments in the western equatorial Pacific under La Niña conditions from January to November 1999. Eleven species contributed about 90% of the total flux in all traps. Two sites (MT1, MT3) in the Western Pacific Warm Pool region (WPWP) were characterized by common occurrences of the species Globigerinoides ruber, Globigerinoides sacculifer, Globigerinoides tenellus, and Neogloboquadrina dutertrei. Site MT5 farther to the east in the equatorial upwelling region had common occurrences of Globigerina bulloides, Globigerinita glutinata, and Pulleniatina obliquiloculata. Very high abundances of G. bulloides and G. glutinata at MT5 indicate that equatorial upwelling (EU) occurred during the 1999 La Niña. The two western sites have similar assemblage compositions, but MT1 ( 135°E) has the highest fluxes (up to  3800 tests m^− 2 day^− 1), whereas MT3 ( 145° E) has fluxes below  2200 tests m^− 2 day^− 1. Relatively high fluxes (up to  3000 tests m^− 2 day^− 1) occur at site MT5 ( 176° E), where upwelling occurred.The differences in faunal composition in the WPWP and EU might be attributable to differences in the way in which nutrients are supplied to the phytoplankton: large amounts of suspended material are supplied to the WPWP by advection of waters passing through the coastal region of an archipelago, whereas upwelling of nutrient-rich waters enhances primary production in the EU. At the westernmost site in the WPWP, a peak in the G. bulloides flux coincided with southward flow of the New Guinea Coastal Current (NGCC) in late February, but the highest G. ruber flux coincided with northward flow of this current in late May. Thus, the differences in species dominance at this location may be caused by monsoon-driven variability in the flow direction of the NGGC.     

Effect of application rates and abiotic factors on Steinernema carpocapsae for control of overwintering navel orangeworm (Lepidoptera: Pyralidae, Amyelois transitella) in pistachios

The effect of reduced application rate, soil temperature at shallow depth (2.5 cm), and soil type on the efficacy of Steinernema carpocapsae against the navel orangeworm, Amyelois transitella, was evaluated in six field trials employing 1 m² plots conducted from November 2003 through December 2004 in Madera and Kern Counties, California. Nematodes were applied at a concentration of 100,000 infective juveniles (IJs)/m² (10⁹/ha) in a volume of 187 ml water/m² (1870 L/ha) with a post-application irrigation in all trials. Mortality ranged from 7.9 to 64.9% in successful trials and percent reduction in live larvae per plot was as high as 74.6%. Percent reduction and mortality were highly correlated (r² = 0.78) and larval reduction typically was 10–11% greater than mortality for any treatment. In one trial, although nematode treatment significantly increased mortality compared to the controls, the treatment was deemed unsatisfactory because mortality was <15%. Soil temperature in this trial rose to 39 °C within 5 h after application. Nematodes failed in two other trials when soil temperature fell below freezing (minimum temperatures −3.0, −5.5 °C, respectively) several times in a 5-day period. We conclude that a commercially feasible application volume of 1870 L water/ha followed by post-application irrigation at this same rate was effective, and that soil maximum temperature at or below 32 °C during the first 24 h after application is necessary for treatment success.     

Trophi morphology and its usefulness for identification of formalin-preserved species of Synchaeta Ehrenberg, 1832 (Rotifera: Monogononta: Synchaetidae)

A study of temporal and spatial distribution of zooplankton frequently requires identification to species level. In such studies, samples are usually fixed, but according to different authors rotifer genera such as Synchaeta (Ehrenberg, 1832) can only be identified in live samples. A procedure for the identification of preserved specimens of Synchaeta is presented, permitting ecological studies without the examination of live material. Trophus morphology, investigated by light and scanning electron microscopy, was related to morphological characteristics of preserved specimens. In this way, body length was used to group formalin-preserved specimens into Synchaeta gr. tremula-oblonga sensu Ruttner-Kolisko (1974) (170 μm) or Synchaeta gr. stylata-pectinata sensu Ruttner-Kolisko (1974) (190 μm). Furthermore, body length, colour and shape were used to identify Synchaeta grandis (Zacharias, 1893), Synchaeta pectinata (Ehrenberg, 1832), Synchaeta kitina (Rousselet, 1902) and Synchaeta lakowitziana (Lucks, 1930). It was, however, impossible to distinguish Synchaeta tremula (Müller, 1786) from Synchaeta oblonga (Ehrenberg, 1832) by their trophi because of contradictions in the diagnostic keys and monographs regarding trophus morphology.     

Whinchat Saxicola rubetra reproductive success according to hay cutting schedule and meadow passerine density in alluvial and upland meadows in France

In France, whinchat Saxicola rubetra populations now rarely breed at altitudes below 1,000 m except in flooded alluvial plains. Whinchat reproductive success, the timing of haymaking and meadow bird density were assessed in 2003 in six areas selected in three different flooded plains, in five areas from two massifs at altitudes ranging from 990 to 1,250 m, and in 2004 in six areas of three flooded plains, in seven areas from three massifs at altitudes ranging from 1,200 to 2,000 m. The percentage of whinchat territories in which juveniles were observed was negatively correlated with mown areas at the time when 80% of hatched broods (observation of prey carryings by adults) would theoretically be fledged. Additionally, assessed hatching success was negatively correlated with early haymaking in 2003 and with meadow passerine territory density in 2004. In upland meadows, whinchat populations exhibited either low reproductive success and low density (1,200 m) or high reproductive success and high density (1,200 m). In lowland flooded meadow however, high density could match low reproductive success and whinchat population then acts as a sink population. We conclude that adequate agri-environmental policy should not focus only on lowland meadows, even if the highest meadow bird diversity is often found there.     

GC/MS-SCAN to follow the fate of crude oil components in bioreactors set to remediate contaminated soil

In this paper GC/MS-SCAN was used to identify the crude oil components that persist after bioremediation treatment of contaminated soil and the metabolites generated during this process. The soil was treated in bioreactors inoculated with an adapted bacterial population. In the first of two sets of conditions used, the water phase of the reactor was circulated continuously at a flow rate of 7 l day⁻¹, and in the other case, it was circulated for a short period once a day to give the equivalent of 0.5 l day⁻¹. Data showed that acyclic, n- and substituted alkanes C₁₂ were still present after one year of remediation, while the majority of substituted derivatives of polycyclic and aromatic hydrocarbons could not be detected by GC/MS-SCAN analysis. The number of components identified was about one-half of that initially observed. After treatment the same number of components was detected at the top and bottom of the reactor in which the water phase was circulated continuously, whereas a smaller number of metabolites were observed at the top rather than at the bottom of the reactor with discontinuous circulation. The analysis has pointed to the importance of n-alkanes, their substituted derivatives and polycyclic aromatic hydrocarbons as the most significant pollutants.     

The control of transmembrane helix transverse position in membranes by hydrophilic residues

The ability of hydrophilic residues to shift the transverse position of transmembrane (TM) helices within bilayers was studied in model membrane vesicles. Transverse shifts were detected by fluorescence measurements of the membrane depth of a Trp residue at the center of a hydrophobic sequence. They were also estimated from the effective length of the TM-spanning sequence, derived from the stability of the TM configuration under conditions of negative hydrophobic mismatch. Hydrophilic residues (at the fifth position in a 21-residue hydrophobic sequence composed of alternating Leu and Ala residues and flanked on both ends by two Lys) induced transverse shifts that moved the hydrophilic residue closer to the membrane surface. At pH 7, the dependence of the extent of shift upon the identity of the hydrophilic residue increased in the order: L < GYT < RH < S < P < K < EQ < N < D. By varying pH, shifts with ionizable residues fully charged or uncharged were measured, and the extent of shift increased in the order: L < GYH^oT < E^oR < S < P < K⁺< QD^oH⁺ < NE^– < D^–. The dependence of transverse shifts upon hydrophilic residue identity was consistent with the hypothesis that shift magnitude is largely controlled by the combination of side chain hydrophilicity, ionization state, and ability to position polar groups near the bilayer surface (snorkeling). Additional experiments showed that shift was also modulated by the position of the hydrophilic residue in the sequence and the hydrophobicity of the sequence moved out of the bilayer core upon shifting. Combined, these studies show that the insertion boundaries of TM helices are very sensitive to sequence, and can be altered even by weakly hydrophilic residues. Thus, many TM helices may have the capacity to exist in more than one transverse position. Knowledge of the magnitudes of transverse shifts induced by different hydrophilic residues should be useful for design of mutagenesis studies measuring the effect of transverse TM helix position upon function.     

Extraction and characterization of hemicelluloses from flax shives by different methods

Hemicelluloses were extracted from flax shives using pressurized low-polarity water (PLPW), pressurized aqueous ethanol (PAE), microwave-assisted water (MW-Water) or aqueous ethanol (MW-EtOH), and precipitated with ethanol. Hemicelluloses still remaining in solution were further separated using ultrafiltration. All samples were characterized with chemical analysis, ion-moderated partition chromatography (IMP), size exclusion chromatography (SEC), and Fourier transform infrared (FT-IR) spectroscopy. PLPW, PAE, MW-Water and MW-EtOH extracted 90, 80, 18, and 40% of the total hemicelluloses, respectively. The molecular weight of the ethanol-precipitated hemicelluloses ranged from approximately 11,000 to 40,000 Da and the ethanol-soluble low-molecular weight hemicelluloses were about 1700 Da. High-molecular weight hemicellulose isolated from PAE extracts contained 23% lignin, while that from the PLPW extracts contained 5% lignin. Low-molecular weight hemicelluloses separated by ultrafiltration from PLPW and PAE extracts contained similar amounts of lignin (20%). However, the yield of low-molecular weight hemicelluloses from PLPW was higher (15%) compared to that from PAE (6%). The FT-IR results revealed the specific band maximum at 1220 cm⁻¹ and the bands between 1175 and 1000 cm⁻¹ which are typical of xylans.