Biodegradation of ballast tank coating investigated by impedance spectroscopy and microscopy.

This research paper addresses the biodegradation process for ballast tank coatings in marine environments. As part of this new approach, a commercially available ballast tank coating was exposed to bacteria obtained from a culture collection and to a natural bacterial community isolated from a real ballast tank. The natural community was chosen to explore the interaction of natural biofilms with the coating, an aspect, which is not covered in standard procedures. It is shown that biological activity significantly affects the coating properties. Micro-cracks and holes have been identified using AFM. Acidic bacteria generated holes with 0.2-0.9 µm in depth and 4-9 µm in width. Whereas the natural community additionally caused cracks of 2-8 µm in depth and 1 µm in length. The overall effect of this degradation was examined using the EIS technique. However, the bacterial affected coatings (exposed to acid producing bacteria and a natural community) show a decrease in corrosion resistance. Impedance IZI values decreased over time from 1.18 × 10(9) to 1.87 × 10(7) Ω for acidic bacteria and from 1.71 × 10(9) to 2.24 × 10(7) Ω for the natural community, indicating a clear loss in coating resistance over time. It is also revealed that the coating corrosion resistance declines after 40 days of exposure for the natural community, leading to the formation of blisters. Bacterial settling could be linked to some specific biofilm patterns affecting different types of coating attack. It can be concluded that it is necessary to include natural communities in coating degradation studies to identify possible degradation mechanisms and the severity of the attack over time.

Responses of antioxidant enzymes to cold and high light are not correlated to freezing tolerance in natural accessions of Arabidopsis thaliana.

Low temperatures and high light cause imbalances in primary and secondary reactions of photosynthesis, and thus can result in oxidative stress. Plants employ a range of low-molecular weight antioxidants and antioxidant enzymes to prevent oxidative damage, and antioxidant defence is considered an important component of stress tolerance. To figure out whether oxidative stress and antioxidant defence are key factors defining the different cold acclimation capacities of natural accessions of the model plant Arabidopsis thaliana, we investigated hydrogen peroxide (H2 O2 ) production, antioxidant enzyme activity and lipid peroxidation during a time course of cold treatment and exposure to high light in four differentially cold-tolerant natural accessions of Arabidopsis (C24, Nd, Rsch, Te) that span the European distribution range of the species. All accessions except Rsch (from Russia) had elevated H2 O2 in the cold, indicating that production of reactive oxygen species is part of the cold response in Arabidopsis. Glutathione reductase activity increased in all but Rsch, while ascorbate peroxidase and superoxide dismutase were unchanged and catalase decreased in all but Rsch. Under high light, the Scandinavian accession Te had elevated levels of H2 O2 . Te appeared most sensitive to oxidative stress, having higher malondialdehyde (MDA) levels in the cold and under high light, while only high light caused elevated MDA in the other accessions. Although the most freezing-tolerant, Te had the highest sensitivity to oxidative stress. No correlation was found between freezing tolerance and activity of antioxidant enzymes in the four accessions investigated, arguing against a key role for antioxidant defence in the differential cold acclimation capacities of Arabidopsis accessions.

Development and use of guideline-derived quality indicators for community lymphoedema.

BACKGROUND: Treatment of lymphoedema is complex and needs specific skills. There are no standards for the evaluation of quality of care. OBJECTIVE: Development and application of indicators for the measurement of quality of care in lymphoedema. METHODS: In a three-step process including a national Delphi expert consensus, quality indicators were derived from national and international guidelines. In a cross-sectional study involving a large spectrum of care providers, the quality of lymphoedema care in the community was assessed by transforming the indicators to one unweighted quality index (QI). RESULTS: A total of 12 quality indicators were identified and applied to n = 348 patients with lymphoedema and lipolymphoedema of any origin in the metropolitan area of Hamburg (90.8% female, mean age 57, SD 14.5 years). On average, 55% of the quality indicators were met, and 64.8% of the patients were satisfied with lymphoedema care. There was a significant correlation between QI and satisfaction. CONCLUSIONS: The quality indicators and the QI are feasible and valid for the evaluation of quality of care. They can support optimizing lymphoedema care.

Trehalose and vitreous states: desiccation tolerance of dormant stages of the crustaceans Triops and Daphnia.

Several aquatic organisms are able to withstand extreme desiccation in at least one of their life stages. This is commonly known as "anhydrobiosis." It was often thought that to tolerate such a desiccated state required high amounts of compatible solutes such as the nonreducing disaccharide trehalose, which protects cellular structures by water replacement and glass formation. Trehalose levels of dormant eggs and cysts of five freshwater crustaceans (Daphnia magna, Daphnia pulex, Triops longicaudatus, Triops cancriformis, and Triops australiensis) were observed in different states of hydration and dehydration. Although trehalose was detected in all species, the concentration was under 0.5% of the dry weight (0.05 µg/µg protein), and no change between the different states was observed. Differential scanning calorimetry (DSC) measurements indicated that dried cysts of all Triops species were in a glassy state, supporting the vitrification hypothesis. No indication for a vitreous state was found in dried resting eggs of Daphnia.

Understanding Participation in a Hospital-Based HIV Support Group in Limpopo Province; South Africa

Background: Support groups are an appropriate way of delivering psychosocial support to people living with HIV/AIDS; especially in low-resource countries. The aim of the study was to understand why people with HIV attended psychosocial support groups. Methods: This was a qualitative study design using focus-group discussions in which support-group members volunteered to participate. Five focus groups were involved in the study. Results: The participants attended because they were referred by a health-care worker; wanted information; wanted emotional support; accompanied an ill relative or knew about the support group. Perceived benefits included receiving psychological support; accepting one's HIV status; reducing stigma and isolation; increasing hope; forging new friendships; helping others; obtaining HIV-related information; developing strategies to change behaviour; gaining access to medical care at the adjoining HIV clinic and receiving food donations. Negative aspects of attending the support group included the large size of the support group; long queues at the HIV clinic; concerns about confidentiality and negative staff attitudes towards the participants. Leaders were concerned about conflict; burn-out and impractical protocols. Access to disability grants was also a concern. Conclusions: Support groups can assist members to cope with the various challenges associated with living with HIV/ AIDS through offering structured emotional; informational; instrumental and material support. Support group sizes should be limited. A structured curriculum containing up-to-date information about ART should also be offered to support groups. Social workers should furthermore be involved to facilitate access to appropriate social grants. Finally; support group leaders should receive appropriate training and regular debriefing

Compensatory growth response in pigs: effects on growth performance, composition of weight gain at carcass and muscle levels, and meat quality.

A restriction/realimentation feeding strategy was applied to pigs to increase the age at market weight and final ADG, modify protein and lipid deposition rates at carcass and muscle levels, and thereby improve eating quality of the pork. A total of 126 Duroc x (Large White x Landrace) pigs (females and castrated males) were used. At the average BW of 30 kg, within litter and sex, pairs of littermates (blocked by BW) were randomly assigned to ad libitum (AL) feeding during growing (30 to 70 kg of BW) and finishing (70 to 110 kg of BW) periods (AL, n = 56), or restricted feeding at 65% of the ADFI of the AL pigs, on a BW basis, during the growing period and AL feeding during finishing (compensatory growth, CG; n = 56). In each feeding regimen, 15 pigs were slaughtered at 70 kg of BW, and 41 pigs were slaughtered at 110 kg of BW. Additionally, 14 pigs were slaughtered at 30 kg of BW to calculate tissue deposition rates. The CG pigs showed decreased ADG (-35%, P = 0.001) during growing but increased ADG (+13%, P = 0.001) during finishing (i.e., compensatory growth) due to greater (P = 0.001) ADFI and G:F. Hence, CG pigs were 19 d older at 110 kg of BW than AL pigs. The CG pigs were leaner at 70 kg of BW than AL (e.g., 11.7 vs. 13.5 mm of average backfat thickness for CG and AL pigs, respectively, P = 0.023), whereas the differences were reduced at 110 kg of BW (20.6 vs. 21.0 mm of average backfat thickness for CG and AL pigs, respectively, P = 0.536). At 70 kg of BW, intramuscular fat (IMF) content of LM did not differ between CG and AL pigs (1.25 vs. 1.49%, respectively, P = 0.118), whereas CG pigs had less IMF in LM at 110 kg of BW (2.19 vs. 2.53% for CG and AL pigs, respectively, P = 0.034). Feeding regimen influenced the composition of weight gain. From 30 to 70 kg of BW, feed restriction reduced (P = 0.001) lean and adipose tissue deposition at the carcass level and protein and lipid deposition at the muscle level. From 70 to 110 kg of BW, the CG feeding strategy increased (P = 0.016) deposition of adipose but not of lean tissue at the carcass level. However, lipid and protein deposition at the muscle level were not affected. Thus, realimentation promoted deposition of subcutaneous fat over IMF. Feeding regimen hardly affected technological meat quality at 110 kg of BW. The CG feeding strategy decreased (P = 0.014) the meat juiciness score in relation to the decreased IMF but did not influence other sensory traits. Elevated IMF content and improved pork quality might be achieved by modifying the onset or duration of the restriction and realimentation periods.

The response of various muscle types to a restriction -re-alimentation feeding strategy in growing pigs.

Muscle lipid concentration is known to influence pork eating quality. This study aimed at evaluating the effect of a restriction-re-alimentation feeding strategy on intramuscular fat deposition in pigs. A total of 70 Duroc × (Large White × Landrace) pigs (castrated males and females) were used. Ten pigs were first slaughtered at 30 kg live weight (LW) to determine initial body and muscle composition. From 30 to 80 kg LW (growing period), pigs were either fed ad libitum (AL) or restricted to 70% of the ad libitum intake of AL pigs (RA). From 80 to 110 kg LW (finishing period), both AL and RA pigs were fed ad libitum. In each group, pigs were slaughtered at 80 kg (n = 10) and at 110 kg (n = 20) LW. During the growing period, the growth rate of RA pigs was reduced by 30% (P < 0.001) compared with AL pigs. During the finishing period, RA pigs had a 7% (P = 0.09) higher growth rate than AL pigs due to compensatory feed intake (+14%). Plasma insulin-like growth factor-1 concentration was lower in RA pigs at 80 kg LW, but markedly increased after re-alimentation up to the level observed in AL pigs (P < 0.001). At 80 kg, the leaner carcasses of RA pigs resulted from a more pronounced reduction in fat than in lean tissue deposition rates. Re-alimentation of RA pigs increased fat tissue deposition (+160% for females, P < 0.01) but not lean deposition in the carcass, leading to limited differences in carcass composition between RA and AL pigs at 110 kg LW. Regarding tissue deposition rates, the response to feeding strategy differs between muscles. In the m. biceps femoris (BF), restriction affected lipid (-50%, P < 0.001) and protein (-25%, P < 0.001) deposition, whereas re-alimentation increased lipid (+62%, P < 0.05) but not protein deposition rates. At market weight, the extent of the difference in BF lipid concentration between RA and AL pigs was strongly reduced, but still significant. By contrast, in the m. longissimus, restriction decreased protein but not lipid deposition, whereas neither of them was modified during re-alimentation. Therefore, an increased muscle lipid concentration at 110 kg LW could not be reached in RA pigs. Modifications of onset and/or duration of restriction and re-alimentation periods should be tested to optimise effects on muscle lipid deposition and thereby achieve improved pork quality.

Adherence To HIV Antiretroviral Therapy Part II: Which Interventions Are Effective In Improving Adherence?

Interventions to support adherence to antiretroviral therapy (ART) can be classified into four categories: cognitive; beha-vioural and affective interventions and (modified) directly observed therapy (DOT.) Cognitive interventions improve HIV- and ART-related knowledge; but this is not consistently associated with better adhe-rence. Cognitive interventions that are combined with behavioural or psychological strategies are more effective in improving adherence; especially in patients who previously were less adherent. These include interventions that improve self-efficacy; provide stress management/expressive support therapy or motivational interviewing. As yet there is no evidence for the role of affective interventions and modified DOT to improve adherence to ART. When designing interventions to address adherence; it should be borne in mind that multi-component interventions are more effective than single-focus interventions. A combination of educational; behavioural and affective components is suggested to ensure optimum adherence.In countries with a high prevalence of HIV; such as South Africa; careful patient preparation; rather than selecting patients based on non-clinical predictors of adherence; seems an appropriate method for scaling up ART. South African guidelines focus on comprehensive adherence support to all patients; with additional support to patients with less than 80adherence. More research on the effectiveness of interventions aimed at improving adherence is urgently needed; especially in develo-ping countries

Gene cloning and functional characterization by heterologous expression of the fructosyltransferase of Aspergillus sydowi IAM 2544.

We have purified a fructosyltransferase from conidia of the inulin-producing fungus Aspergillus sydowi IAM 2544 and obtained peptide sequences from proteolytic fragments of the protein. With degenerated primers, we amplified a PCR fragment that was used to screen a cDNA library. The fructosyltransferase gene from Aspergillus sydowi (EMBL accession no. AJ289046) is expressed in conidia, while no expression could be detected in mycelia by Northern blot analysis of mycelial RNA. The gene encodes a protein with a calculated molecular mass of 75 kDa that is different from all fructosyltransferases in the databases. The only homology that could be detected was to the invertase of Aspergillus niger (EMBL accession no. L06844). The gene was functionally expressed in Escherichia coli, yeast, and potato plants. With protein extracts from transgenic bacteria and yeast, fructooligosaccharides could be produced in vitro. In transgenic potato plants, inulin molecules of up to 40 hexose units were synthesized in vivo. While in vitro experiments with protein extracts from conidia of Aspergillus sydowi yielded the same pattern of oligosaccharides as extracts from transformed bacteria and yeast, in vivo inulin synthesis with fungal conidia leads to the production of a high-molecular-weight polymer.

Non-disaccharide-based mechanisms of protection during drying.

Few tissues or organisms can survive the removal of nearly all their intra and extracellular water. These few have developed specialized adaptations to protect their cellular components from the damage caused by desiccation and rehydration. One mechanism, common to almost all such organisms, is the accumulation of disaccharides within cells and tissues at the onset of dehydration. This adaptation has been extensively studied and will not be considered in this review. It has become increasingly clear that true desiccation tolerance is likely to involve several mechanisms working in concert; thus, we will highlight several other important and complimentary adaptations found especially in the dehydration-resistant tissues of higher plants. These include the scavenging of reactive oxygen species, the down-regulation of metabolism, and the accumulation of certain amphiphilic solutes, proteins, and polysaccharides.

Survival of hippocampal neurons in culture upon hypoxia: effect of erythropoietin.

The potential of erythropoietin (EPO) to reduce hypoxia-induced cell death has been investigated in 5-day-old primary cultures of rat postnatal hippocampal neurons. Application of EPO (100 pM) at the start of hypoxia resulted in a significant reduction of neuronal death (33.0 +/- 7.5% in cells incubated with EPO vs 56.75 +/- 7.3% in non-treated cells; n = 4, p < 0.021). Similiar results were obtained upon application of cycloheximide (CHX; 1 microM) simultaneously with hypoxia (34.75 +/- 5.6% vs 56.75 +/- 7.3% with and without CHX, respectively, n = 4, p < 0.035), indicating that hypoxia-induced neuronal death is an active, protein synthesis-dependent process. Both, EPO and EPO receptor (EPOR) were found to be expressed after hypoxia in hippocampal neurons in vitro and in vivo. These results demonstrate for the first time that EPO can reverse hypoxia-induced neuronal death when applied simultaneously with the hypoxic stimulus.

Transgenic potato (Solanum tuberosum) tubers synthesize the full spectrum of inulin molecules naturally occurring in globe artichoke (Cynara scolymus) roots.

The ability to synthesize high molecular weight inulin was transferred to potato plants via constitutive expression of the 1-SST (sucrose:sucrose 1-fructosyltransferase) and the 1-FFT (fructan: fructan 1-fructosyltransferase) genes of globe artichoke (Cynara scolymus). The fructan pattern of tubers from transgenic potato plants represents the full spectrum of inulin molecules present in artichoke roots as shown by high-performance anion exchange chromatography, as well as size exclusion chromatography. These results demonstrate in planta that the enzymes sucrose:sucrose 1-fructosyltransferase and fructan:fructan 1-fructosyltransferase are sufficient to synthesize inulin molecules of all chain lengths naturally occurring in a given plant species. Inulin made up 5% of the dry weight of transgenic tubers, and a low level of fructan production also was observed in fully expanded leaves. Although inulin accumulation did not influence the sucrose concentration in leaves or tubers, a reduction in starch content occurred in transgenic tubers, indicating that inulin synthesis did not increase the storage capacity of the tubers.

Plant fructans stabilize phosphatidylcholine liposomes during freeze-drying.

Fructans have been implicated as protective agents in the drought and freezing tolerance of many plant species. A direct proof of their ability to stabilize biological structures under stress conditions, however, is still lacking. Here we show that inulins (linear fructose polymers) isolated from chicory roots and dahlia tubers stabilize egg phosphatidylcholine large unilamellar vesicles during freeze-drying, while another polysaccharide, hydroxyethyl starch, was completely ineffective. Liposome stability was assessed after rehydration by measuring retention of the soluble fluorescent dye carboxyfluorescein and bilayer fusion. Inulin was an especially effective stabilizer in combination with glucose. Analysis by HPLC showed that the commercial inulin preparations used in our study contained no low molecular mass sugars that could be responsible for the observed stabilizing effect of the fructans. Fourier transform infrared spectroscopy showed a reduction of the gel to liquid-crystalline phase transition temperature of dry egg PtdCho by more than 20 degrees C in the presence of inulin. A direct interaction of inulin with the phospholipid in the dry state was also indicated by dramatic differences in the phosphate asymmetric stretch region of the infrared spectrum between samples with and without the polysaccharide.

Arabidopsis knockout mutation of ADC2 gene reveals inducibility by osmotic stress.

We isolated an Arabidopsis thaliana mutant line carrying an insertion of the En-1 transposable element at the ADC2 locus. The insertion causes a knockout of the arginine decarboxylase 2 gene. We demonstrated that ADC2 is the gene responsible for induction of the polyamine biosynthetic pathway by osmotic stress. No induction of ADC activity by the osmolite sorbitol could be observed in the homozygous mutant, indicating a predominant role of ADC2 in stress response. ADC activity is reduced in the mutant by 44% under non-stressed conditions and the mutant shows no obvious phenotype. This is the first report of a genetically mapped mutation in the polyamine biosynthetic pathway in plants.

Production of modified polymeric carbohydrates.

The cloning of a gene responsible for the phosphorylation of glucans has made it possible to genetically engineer the phosphorylation level of starches in higher plants. Through the manipulation of starch synthase activity, it is now also possible to genetically tailor the chain-length distribution in the amylopectin. Both findings will lead to the development of novel starches utilized as a renewable resource. The production of fructans on a large scale can also be envisioned for the near future.

Differences in chain length distribution of inulin from Cynara scolymus and Helianthus tuberosus are reflected in a transient plant expression system using the respective 1-FFT cDNAs.

A newly isolated cDNA clone, Cy3, encoding the fructan fructan 1-fructosyltransferase (1-FFT) from artichoke was expressed using tobacco protoplasts as expression system. Analysis of the inulin molecules synthesized upon incubation of protoplast extracts with a mixture of oligofructans (DP3-5) shows the production of inulins with a degree of polymerization (DP) of up to 23, whereas parallel experiments performed using a 1-FFT cDNA from Jerusalem artichoke led to the production of fructans with a DP of up to only 12. The results of in vitro fructan synthesis catalyzed by transiently expressed enzymes therefore reflect the difference of in vivo fructan composition of Jerusalem artichoke (M(DP) = 8-10) and artichoke (M(DP) = 65). These data suggest that the fructan pattern in a given species is mainly defined by the enzymatic characteristics of 1-FFT.

Production of 1-kestose in transgenic yeast expressing a fructosyltransferase from Aspergillus foetidus.

Sucrose-inducible secretory sucrose:sucrose 1-fructosyltransferase (1-SST) from Aspergillus foetidus has been purified and subjected to N-terminal amino acid sequence determination. The enzyme is extensively glycosylated, and the active form is probably represented by a dimer of identical subunits with an apparent molecular mass of 180 kDa as judged from mobility in seminative acrylamide gels. The enzyme catalyzes fructosyl transfer from sucrose to sucrose producing glucose and 1-kestose. Oligosaccharides with a higher degree of polymerization are not obtained with sucrose as the substrate. The cDNA encoding the A. foetidus 1-SST has been cloned and sequenced. Sequence homology was found to be highest to levanases, but no hydrolytic activity was observed when levan was incubated with the enzyme. Expression of the cloned gene in an invertase-deficient mutant of Saccharomyces cerevisiae resulted in 1-kestose production, with 6-kestose and neokestose being side products of the reaction. Products were well distinguishable from those formed by yeast transformants expressing a cytosolic invertase.

Structure of the enzymatically synthesized fructan inulin.

Construction, purification and characterization of a fusion protein of maltose-binding protein of Escherichia coli and the fructosyltransferase of Streptococcus mutans is described. With the purified protein, in vitro synthesis of inulin was performed. The obtained polysaccharide was characterized by high-performance size-exclusion chromatography (HPSEC) and static light scattering (SLS) in dilute aqueous and dimethyl sulfoxide solution. For all samples very high molecular weights between 60 x 10(6) and 90 x 10(6) g/mol and a remarkable small polydispersity index of 1.1 have been determined. Small root-mean-square radii of gyration point to a compact conformation in dilute solution. No difference between native and enzymatically synthesized inulin was observed by X-ray powder diffraction and thermoanalysis of solid samples.

Increased potato tuber size resulting from apoplastic expression of a yeast invertase.

The role of sucrose cleavage in determining sink strength in potato was investigated by generating transgenic potato plants that expressed a yeast invertase in either the cytosol or apoplast of tubers. Cytosolic localization gave rise to a reduction in tuber size and an increase in tuber number per plant whereas apoplastic targeting led to an increase in tuber size and a decrease in tuber number per plant. Sink organ size can be manipulated through modification of sucrose metabolism.

Transgenic potato tubers accumulate high levels of 1-kestose and nystose: functional identification of a sucrose sucrose 1-fructosyltransferase of artichoke (Cynara scolymus) blossom discs.

By screening a cDNA library of artichoke (Cynara scolymus) blossom discs for fructosyltransferases, we isolated a clone designated Cy21. The deduced amino acid sequence shows homology to acid beta-fructosyl hydrolases and to the sucrose-fructan 6-fructosyltransferase (6-SFT) of barley. Transiently expressed in Nicotiana tabacum protoplasts, the Cy21 gene-product synthesized 1-kestose, indicating that Cy21 codes for a sucrose sucrose 1-fructosyltransferase (1-SST). The enzyme worked at physiologically relevant sucrose concentrations (25 mM sucrose). In the protoplast system, 1-kestose seemed to be the only fructan product of the 1-SST. The enzyme activity was not affected by pyridoxal-HCl, an inhibitor of both the beta-fructosyl hydrolase and the fructosyltransferase activity of invertases. The fructosyltransferase activity of the Cy21 gene-product, however, could be inhibited by Zn2+, Ag+ and Cu2+ ions. In artichoke plants the Cy21 transcript was highly abundant in primary roots and blossom discs. Transgenic potato tubers expressing Cy21 contain high levels of 1-kestose along with nystose and traces of fructosyl-nystose, supporting the conclusion that the Cy21 clone encodes a sucrose sucrose 1-fructosyltransferase.