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Evolutionary diversity can be driven by the interaction of plants with different environments. Molecular bases involved in ecological adaptations to abiotic constraints can be explored using genomic tools. Legumes are major crops worldwide and soil salinity is a main stress affecting yield in these plants. We analyzed in the Medicago truncatula legume the root transcriptome of two genotypes having contrasting responses to salt stress: TN1.11, sampled in a salty Tunisian soil, and the reference Jemalong A17 genotype. TN1.11 plants show increased root growth under salt stress as well as a differential accumulation of sodium ions when compared to A17. Transcriptomic analysis revealed specific gene clusters preferentially regulated by salt in root apices of TN1.11, notably those related to the auxin pathway and to changes in histone variant isoforms. Many genes encoding transcription factors (TFs) were also differentially regulated between the two genotypes in response to salt. Among those selected for functional studies, overexpression in roots of the A17 genotype of the bHLH-type TF most differentially regulated between genotypes improved significantly root growth under salt stress. Despite the global complexity of the differential transcriptional responses, we propose that an increase in this bHLH TF expression may be linked to the adaptation of M. truncatula to saline soil environments.

Legume crops related to the model plant Medicago truncatula can adapt their root architecture to environmental conditions, both by branching and by establishing a symbiosis with rhizobial bacteria to form nitrogen-fixing nodules. Soil salinity is a major abiotic stress affecting plant yield and root growth. Previous transcriptomic analyses identified several transcription factors linked to the M. truncatula response to salt stress in roots, including NAC (NAM/ATAF/CUC)-encoding genes. Over-expression of one of these transcription factors, MtNAC969, induced formation of a shorter and less-branched root system, whereas RNAi-mediated MtNAC969 inactivation promoted lateral root formation. The altered root system of over-expressing plants was able to maintain its growth under high salinity, and roots in which MtNAC969 was down-regulated showed improved growth under salt stress. Accordingly, expression of salt stress markers was decreased or induced in MtNAC969 over-expressing or RNAi roots, respectively, suggesting a repressive function for this transcription factor in the salt-stress response. Expression of MtNAC969 in central symbiotic nodule tissues was induced by nitrate treatment, and antagonistically affected by salt in roots and nodules, similarly to senescence markers. MtNAC969 RNAi nodules accumulated amyloplasts in the nitrogen-fixing zone, and were prematurely senescent. Therefore, the MtNAC969 transcription factor, which is differentially affected by environmental cues in root and nodules, participates in several pathways controlling adaptation of the M. truncatula root system to the environment., (© 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd.)

Health care workers (HCW) are at increased risk for acquisition of hepatitis B virus (HBV) infection from occupational exposure. This can be prevented by active immunization. We performed a retrospective chart review of HCW who were persistent low (anti-HBs antibody values <100 U/L) or non responders (<10 U/L) after 6 active immunizations and demonstrate successful immunization (anti-HBs ≥100 U/l) after a total of 8-14 vaccine doses in 13 such HCW by use of various vaccination schedules. This "proof of principle" should encourage occupational health care providers to convince HCW to accept further vaccine doses until the targeted threshold considered to be the correlate of immunity has been reached. Prospective studies should be performed to determine the optimal schedule of further booster doses for HCW who are persistent non or low responders.

The adaptation of root architecture to environmental constraints is a major agricultural trait, notably in legumes, the third main crop worldwide. This root developmental plasticity depends on the formation of lateral roots (LRs) emerging from primary roots. In the model legume Medicago truncatula, the HD-Zip I transcription factor HB1 is expressed in primary and lateral root meristems and induced by salt stress. Constitutive expression of HB1 in M. truncatula roots alters their architecture, whereas hb1 TILLING mutants showed increased lateral root emergence. Electrophoretic mobility shift assay, promoter mutagenesis, and chromatin immunoprecipitation-PCR assays revealed that HB1 directly recognizes a CAATAATTG cis-element present in the promoter of a LOB-like (for Lateral Organ Boundaries) gene, LBD1, transcriptionally regulated by auxin. Expression of these genes in response to abscisic acid and auxin and their behavior in hb1 mutants revealed an HB1-mediated repression of LBD1 acting during LR emergence. M. truncatula HB1 regulates an adaptive developmental response to minimize the root surface exposed to adverse environmental stresses.

The only treatment of end-stage renal disease patients undergoing chronic dialysis is kidney transplantation. However, about half of graft recipients encounter organ loss within ten years after renal transplantation. There is emerging evidence that the presence of alloreactive antibodies against non-HLA antigens in the serum of the recipient prior transplantation is associated with higher incidence of chronic rejection. However, the molecular identity of these antigens is largely unknown. To determine the most common non-HLA antigens, we tested lymphocytic extracts from 20 healthy volunteers with sera of 28 patients on the transplantation waiting list by Western blotting. There was a group of five proteins that was recognized by most sera. Using patient's own lymphocytes revealed that autoimmunity plays a minor role in this recognition. Two-dimensional Western blotting experiments followed by mass spectrometry identified the antigens as tubulin beta chain, vimentin, lamin-B1, and Rho GDP-dissociation inhibitor 2. A detailed analysis of vimentin expression revealed that the antigenic 60 kDa isoform is underrepresented in patient's lymphocytes in comparison to those of healthy volunteers. The study revealed that preformed alloreactive antibodies are directed against a small number of specific protein isoforms. Our findings could provide a basis for future improvement of donor-recipient matching.

Background: Cocaine dependence is a major public health problem for which there is no FDA-approved pharmacological treatment. Baclofen is a GABA(B) receptor agonist that in preclinical and early pilot clinical trials has shown promise for the treatment of cocaine dependence. The purpose of this multi-site, double-blind study, was to compare the safety and efficacy of baclofen (60 mg/day) vs placebo in an 8-week treatment of individuals with severe cocaine dependence. The primary outcome measure was subjects' self-reported cocaine use substantiated by urine benzoylecgonine (BE). Analysis of the data did not show a significant difference between the groups treated with baclofen and placebo. The current results do not support a role for 60 mg baclofen in treating cocaine dependence in the population studied. The contrast of this result to earlier, preclinical and human pilot data with baclofen may reflect the trial's focus on severe cocaine-dependent users, and/or the need for a higher baclofen dose. Baclofen's potential as a relapse prevention agent was not tested by the current design, but may be a useful target for future studies.

The root apex contains meristematic cells that determine root growth and architecture in the soil. Specific transcription factor (TF) genes in this region may integrate endogenous signals and external cues to achieve this. Early changes in transcriptional responses involving TF genes after a salt stress in Medicago truncatula (Mt) roots were analysed using two complementary transcriptomic approaches. Forty-six salt-regulated TF genes were identified using massive quantitative real-time RT-PCR TF profiling in whole roots. In parallel, Mt16K+ microarray analysis revealed 824 genes (including 84 TF sequences) showing significant changes (p < 0.001) in their expression in root apexes after a salt stress. Analysis of salt-stress regulation in root apexes versus whole roots showed that several TF genes have more than 30-fold expression differences including specific members of AP2/EREBP, HD-ZIP, and MYB TF families. Several salt-induced TF genes also respond to other abiotic stresses as osmotic stress, cold and heat, suggesting that they participate in a general stress response. Our work suggests that spatial differences of TF gene regulation by environmental stresses in various root regions may be crucial for the adaptation of their growth to specific soil environments.

The disposal of coal combustion residues (CCR) has led to a significant consumption of land in the West Balkan region. In Tuzla (Bosnia and Herzegovina) we studied previously soil-covered (farmed) and barren CCR landfills including management practises, field ageing of CCR and the transfer of trace elements into crops, wild plants and wastewaters. Soil tillage resulted in mixing of cover soil with CCR. Medicago sativa showed very low Cu:Mo ratios (1.25) which may cause hypocuprosis in ruminants. Total loads of inorganic pollutants in the CCR transport water, but not pH ( approximately 12), were below regulatory limits of most EU countries. Arsenic concentrations in CCR transport water were <2microgl(-1) whereas reductive conditions in an abandoned landfill significantly enhanced concentrations in leachates (44microgl(-1)). The opposite pattern was found for Cr likely due to large initial leaching of CrVI. Public use of landfills, including farming, should be based on a prior risk assessment due to the heterogeneity of CCR.

This clinical trial evaluated a contingency management intervention designed to improve medication adherence among HIV-positive methadone maintenance patients. After a 4-week baseline observation phase, eligible participants (N=66) were randomly assigned to: (a) medication coaching sessions every other week to assist with adherence strategies (comparison group) or (b) medication coaching plus voucher reinforcement for opening electronic medication caps on time (voucher group). Baseline adherence (percent doses taken/percent total possible doses) was 51% using electronic measurement, 75% using self-report and 75% using pill count. The intervention was provided for 12 weeks, with a 4-week follow-up. The primary outcome results of the clinical trial indicated effectiveness during the intervention, with significant mean adherence differences between voucher and comparison groups using electronic measurement (78% versus 56%), pill count (86% versus 75%), and self-report (87% versus 69%). Differences between groups faded after vouchers were discontinued. Contingency management shows promise as a strategy to promote antiretroviral medication adherence in this population.

The aim, methods, and results of long-term low-level measurements of 3H, 7Be, 137Cs and 210Pb in water and sediment of the Austrian part of the Danube are presented. The results are discussed focused on recent and obvious future applications in applied radioecology and environmental research. The long-term radiometric data supported by the low-level monitoring program act as basis for ecological modelling and environmental assessment.

The proximal region of the high-molecular-weight glutenin promoter of the Dx5 gene (PrHMWG-Dx5) carries an atypical bifactorial endosperm box containing two cis-acting elements, namely a G-box like motif followed by a prolamin-box motif (Pb1). Transient expression assays in maize endosperm indicate that a promoter fragment containing at least the G-box like element is necessary and sufficient for maximal expression of the HMWG-Dx5 promoter. In transformed maize, we have shown that a 89 bp sequence bearing the bifactorial endosperm box behaves like a functional cis-acting unit. Its repetition in tandem confers a strong specific additive effect specifically in endosperm tissue. In contrast, the fusion of the activation sequences 1 (as-1) and 2 (as-2) of the cauliflower mosaic virus (CaMV) 35S promoter with HMWG-Dx5 derived promoter sequences deregulates its activity in transformed maize. By gel mobility shift assays we have demonstrated that the G-box like motif may alternatively bind two protein groups which have the same DNA-binding affinities as the transcription factors of either the Opaque2 (O2) family and/or the ASF-1 family.

Fat digestion in humans and some mammals such as dogs requires the successive intervention of two lipases: gastric lipase, which is stable and active despite the highly acidic stomach environment, followed by the classical pancreatic lipase secreted into the duodenum. We previously solved the structure of recombinant human gastric lipase (HGL) at 3.0-A resolution in its closed form; this was the first structure to be described within the mammalian acid lipase family. Here we report on the open structure of the recombinant dog gastric lipase (r-DGL) at 2.7-A resolution in complex with the undecyl-butyl (C11Y4) phosphonate inhibitor. HGL and r-DGL show 85.7% amino acid sequence identity, which makes it relevant to compare the forms from two different species. The open r-DGL structure confirms the previous description of the HGL catalytic triad (Ser(153), His(353), and Asp(324)) with the catalytic serine buried and an oxyanion hole (NH groups of Gln(154) and Leu(67)). In r-DGL, the binding of the C11Y4 phosphonate inhibitor induces part of the cap domain, the lid, to roll over the enzyme surface and to expose a catalytic crevice measuring approximately 20 x 20 x 7 A(3). The C11Y4 phosphonate fits into this crevice, and a molecule of beta-octyl glucoside fills up the crevice. The C11Y4 phosphonate inhibitor and the detergent molecule suggest a possible binding mode for the natural substrates, the triglyceride molecules.

The promoter of the Arabidopsis thaliana L. AtEm1 gene encoding a late embryogenesis abundant protein was fused to the beta-glucuronidase reporter gene and introduced into Brassica napus. The promoter is highly active in the vascular tissues of embryo and pollen grains and also active in petals, sepals, caulinar leaves, and carpels.

We have isolated and sequenced a 9.5 kb genomic region from A. thaliana, located on chromosome 2, which contains two tandemly arranged closely related genes (AtM10 and AtM17) coding for a new family of LEA proteins. The deduced proteins have a molecular mass of 11 and 29 kDa, respectively, are extremely hydrophilic except at their N-termini and share 70% amino acid (aa) identity. A 47 aa motif containing a 6-cysteine domain is present once in AtM10 and four times in AtM17. The short intergenic region, the identical position of the intron and the overall sequence homology suggest that these two genes evolved through a duplication event. This conclusion is supported by the presence of two homologous strictosidine synthase-like (pseudo)genes downstream from AtM17 and AtM10. Expression studies, using AtM10 and AtM17 cDNAs, revealed that both transcripts accumulate exclusively in seeds from late embryogenesis until two days after imbibition. Expression of both genes in young seedlings is repressed during ABA, salt or drought treatment, whereas a cold stress induces the expression of AtM17 only. In situ hybridization revealed that AtM10 transcripts are detected throughout the embryo while those of AtM17 are more localized to cotyledon cells.

Alcoholism and tuberculosis represent very difficult problem in treatment because both illnesses require complex treatment which cannot be adequately obtained on classic wards. Tuberculosis in the last 50 years showed the constant decline of incidence--in Croatia (4,500,000 inhabitants) from approximately 20,000 to approximately 2000 cases per year. This study compares preliminary results of AUDIT (Alcohol Use Disorders Identification Test) in the group of tuberculosis patients with the diagnosis of alcoholism (group 1) and in the group of tuberculosis patients who had not the diagnosis of alcoholism (group 2) and who all were admitted to the Hospital for pulmonary diseases and tuberculosis Klenovnik where is an organised unique ward management for tuberculosis and alcoholic patients functioning for the last 25 years on the basic principles of therapeutic community. Their average AUDIT score was 25.44--very high, and all patients scored above 8, what indicated on harmful and hazardous drinking. Additionally, in the group of patients admitted for hospital treatment of tuberculosis without diagnosis alcoholism there were more than 50% of harmful or hazardous alcohol drinkers who ranged 8 or more on AUDIT score, what could indicate that modern identification of hazardous drinking (for example using AUDIT) should be used in all pulmonary and tuberculosis wards. Patients for whom diagnostic procedure shows that they drink harmfully or hazardously should be treated simultaneously for tuberculosis and alcohol use disorders. Because of hidden alcohol problems the group sociotherapy should be organised for all patients at all wards for treatment of tuberculosis in a common and special alcohologic way.

Production and characterization of human lactoferrin (hLf) in transgenic tobacco is reported. We have engineered two constructs containing either the native signal peptide from human lactoferrin or the signal peptide from sweet potato sporamin fused to human lactoferrin encoding cDNA. N-terminal sequences of rhLf purified from tobacco were identical to Lf from human milk for both constructs. The tobacco rhLf presents a molecular mass closely identical to native protein. Overall sugar composition shows the presence of plant specific xylose while sialic acid is absent. Binding parameters of the recombinant molecule to both Jurkat lymphoblastic T-cells or HT29-18-C1 enterocytes are similar to those of human lactoferrin isolated from milk.

Three models of transitional residential community care for substance abuse patients are defined on the basis of the differential provision of health and treatment services: a psychosocial model, a supportive rehabilitation model, and an intensive treatment model. Facilities that provided a high level of on-site health and treatment services were categorized as following an intensive treatment model; these facilities had the strongest emphasis on medical, dual diagnosis, and family treatment orientations. These facilities also had clearer policies and provided their residents more opportunities to participate in setting policies; however, staff were not more accepting of patient impairment or problem behavior. Patients in facilities with intensive treatment programs obtained more outpatient mental health after care. In each of the three types of facilities, a longer length of care was associated with a lower readmission rate; patients who dropped out of psychosocial model facilities had especially high readmission rates. Overall, the findings identify substantial variations among community residential facilities for substance abuse patients, but show relatively little matching of patients' needs with facility services and programs.

Human utilization of recombinant proteins of therapeutical interest, as hemoglobin, implies that the transgenic host allows a low cost production of the active proteins with minimal risks of pathogen contamination. In this regard, the use of transgenic plants could be of great interest. In particular, the systems based on plants could be one of the most economical transgenic system, compared with the others, because biomass obtention in fields is not expensive.

We tested and expanded Fazio's hypothesis that direct experience enhances attitude-behavior consistency (Fazio & Zanna, 1981) to identify effects of experience on the relation of behavior to subjective norm and intention. We also monitored the endurance of the experience effect over time. In order to manipulate experience, we instructed male college students in the United States to perform the testicle self-exam (TSE) a prescribed number of times during an initial week. We compared attitude, subjective norm, intention, and self-reported behavior across experience condition and three postmanipulation times. Direct experience increased later reported TSE behavior and tended to increase attitude and intention. It also enhanced consistency of attitude, subjective norm, and intention with early reported behavior as well as intention with later reported behavior.

Gas-liquid chromatographic and mass, nuclear magnetic resonance, and infrared spectrometric techniques were utilized to identify some of the metabolites of cyproheptadine in the urine of human subjects who had ingested radiolabeled drug. Aromatic ring hydroxylation (followed by glucuronide conjugation), N-demethylation, and heterocyclic ring oxidation were shown to occur in man. The principal metabolite, however, was identified tentatively as a quaternary ammonium, glucuronide-like conjugate of cyproheptadine. No evidence was found for metabolic changes at the tricyclic ethylene bridge in this species.

The incubation of multiply labeled (2H, 3H, 13C, 14C) cambendazole and glutathione with hepatic microsomes from phenobarbital-dosed hamsters results in the formation of polar metabolites. The major metabolite has been characterized by a variety of isotopic, spectrometric, chromatographic, and degradative/synthetic techniques as a glutathione conjugate of cambendazole in which substitution is on the 4-position of the benzimidazole nucleus. The same metabolite is produced by hepatic microsomes from the rat.

Dimethylformamide dialkylacetals have been found to react readily with primary sulfonamides to form N-dimethylaminomethylene derivatives. These compounds possess excellent gas-liquid chromatographic properties and can be conveniently prepared at the submicrogram level. Their retention times are much greater than those of other sulfonamide derivatives (e.g., N,N-dimethyl) but their ease of preparation and lack of absorptive properties make them attractive for gas-liquid chromatographic studies. The practical applicability of this derivatization approach to biological studies is illustrated by the gas-liquid chromatographic determination of 3-bromo-5-cyanobenzenesulfonamide in ovine blood using 3,5-dibromobenzenesulfonamide as the internal standard. The method has a detection limit of 25 ppb with electron capture detection.

Timolol [3-(3-tert.-butylamino-2-hydroxypropoxy)-4-morpholino-1,2,5-thiadiazole], was rapidly absorbed, metabolized, and effectively excreted in man, rats, and dogs. Peak plasma levels of timolol-14C were observed in these species 1-2 hr after oral administration. Generally, less than 20% of the radioactivity was present in the plasma in the unmetabolized form. The intact drug had a plasma half-life of 28 min in the rat, 48 min in the dog, and 5.5 hr in man. After oral administration of timolol-14C to humans approximately 72% of the dose was excreted in 84 hr, with 66% in the urine and 6% in the feces. In the rat, 58% of an oral dose was excreted in the urine and 26% in the feces. The dog excreted 68% of an oral dose in the urine and 19% in feces in 72 hr. Following intravenous administration, rats excreted 50% in the urine and 28% in the feces, which suggests that extensive biliary excretion occurred. Timolol was extensively metabolized. Approximately 50% of the radioactivity was identified in dog urine as the lactic acid metabolite. An additional metabolite was tentatively identified as the 3-oxomorpholino derivative of timolol. Approximately 20% of the dose in man was excreted in the urine unchanged. Two metabolites, resulting from cleavage of the morpholine ring, were identified as 1-tert-butylamino-3-[4-(2-hydroxyethylamino)-1,2,5-thiadiazol-3-xloxyl-2-propanol, accounting for 10% of the urine radioactivity, and t-tert-butylamino-[4-(N-2-hydroxyethylglycolamido)-1,2,5-thiadiazol-3-yloxy]-2-propanol, accounting for 30%. A minor metabolite, resulting from hydroxylation of a terminal methyl group, accounted for an additional 3% of the urine radioactivity.

GLC with electron-capture detection was successfully applied to the analysis of hydrochlorothiazide in human blood and plasma with a sensitivity (0.05 mug/ml) suitable for use with persons on therapeutic dosage levels. On-column methylation with trimethylanilinium hydroxide in methanol was employed to convert hydrochlorothiazide and its bromo analog, which served as the internal standard, to their tetramethyl derivatives.