Candida albicans PALS-green fluorescent protein (GFP) reporter strains were inoculated into mice in a disseminated candidiasis model, and GFP production was monitored by immunohistochemistry and reverse transcription-PCR (RT-PCR). GFP production from the ALS1 and ALS3 promoters was detected immunohistochemically. ALS1, ALS2, ALS3, ALS4, and ALS9 transcription was detected by RT-PCR, further identifying ALS genes expressed in this model.

Most of the adrenal tumors that are incidentally detected are benign adenomas. The incidence of malignant adrenal tumors including adrenocortical carcinoma (ACC) and primary adrenal lymphoma (PAL) is rather low. As many patients with ACC and PAL are diagnosed at an advanced stage of disease, the overall survival time of both entities remains poor. The therapeutic strategies for both entities differ. Thus an early differentiation between ACC and PAL is necessary. Unfortunately hitherto preoperative diagnosis of potentially malignant adrenal masses is still a main problem in the treatment of adrenal tumors. We present the case of a 57-year-old male patient with ACC and the case of an 87-year-old male patient with PAL and provide a systematic comparison of the clinical and pathological features of both entities. In both cases clinical and radiological features resulted in an initially false diagnosis. Primary surgical therapy was performed in both patients. The patient with PAL died five months after initial surgery. The patient with ACC showed tumor progression with local and systemic recurrence despite adjuvant therapy with mitotane and additional surgical therapy. Prognosis of patients with ACC and PAL seems to be dependant on the ability to start accurate treatment without any time delay. We propose some guidelines for diagnosis and surgical management of adrenal tumors.

The origins and rationale of the Cambridge Neuropsychological Test Automated Battery (CANTAB) as a cross-species translational instrument suitable for use in human neuropsychopharmacological studies are reviewed. We focus on its use for the early assessment and detection of Alzheimer's disease, in particular the Paired Associates Learning (PAL) test. We consider its psychometric properties, neural validation, and utility, including studies on large samples of healthy volunteers, patients with mild cognitive impairment (MCI), and Alzheimer's disease. We demonstrate how it can be applied in cross-species studies using experimental animals to bridge the cross-species translational 'gap'. We also show how the CANTAB PAL has bridged a second translational 'gap' through its application to the early detection of memory problems in primary care clinics, using iPad technology.

Two studies examined children's thought patterns in relation to their responses to social challenge. In Study 1, 4th and 5th graders tried out for a pen pal club under either a performance goal (stressing the evaluative nature of the tryout) or a learning goal (emphasizing the potential learning opportunities). In their behavior and attributions following rejection, children who were focused on a performance goal reacted with more helplessness, whereas children given a learning goal displayed a more mastery-oriented response. Study 2 found that in response to hypothetical socially challenging situations, 4th, 5th, and 6th graders who believed personality was nonmalleable (entity theorists) vs. malleable (incremental theorists) were more likely to endorse performance goals. Together, these studies indicate that children's goals in social situations are associated with their responses to social failure and are predicted by their implicit theories about their personality.

A new, phylogenetically distinct, dissimilatory, Fe(III)-reducing bacterium was isolated from surface sediment of a hydrocarbon-contaminated ditch. The isolate, designated strain PAL-1, was an obligately anaerobic, non-fermentative, motile, gram-negative vibrio. PAL-1 grew in a defined medium with acetate as electron donor and ferric pyrophosphate, ferric oxyhydroxide, ferric citrate, Co(III)-EDTA, or elemental sulfur as sole electron acceptor. PAL-1 also used proline, hydrogen, lactate, propionate, succinate, fumarate, pyruvate, or yeast extract as electron donors for Fe(III) reduction. It is the first bacterium known to couple the oxidation of an amino acid to Fe(III) reduction. PAl-1 did not reduce oxygen, Mn(IV), U(VI), Cr(VI), nitrate, sulfate, sulfite, or thiosulfate with acetate as the electron donor. Cell suspensions of PAL-1 exhibited dithionite-reduced minus air-oxidized difference spectra that were characteristic of c-type cytochromes. Analysis of the 16S rRNA gene sequence of PAL-1 showed that the strain is not related to any of the described metal-reducing bacteria in the Proteobacteria and, together with Flexistipes sinusarabici, forms a separate line of descent within the Bacteria. Phenotypically and phylogenetically, strain PAl-1 differs from all other described bacteria, and represents the type strain of a new genus and species, Geovibrio ferrireducens.

Much remains unknown about how transcription factors and sugars regulate phenylpropanoid metabolism in tuber crops like potato (Solanum tuberosum). Based on phylogeny and protein similarity to known regulators of phenylpropanoid metabolism, 15 transcription factors were selected and their expression was compared in white, yellow, red, and purple genotypes with contrasting phenolic and anthocyanin profiles. Red and purple genotypes had increased phenylalanine ammonia lyase (PAL) enzyme activity, markedly higher levels of phenylpropanoids, and elevated expression of most phenylpropanoid structural genes, including a novel anthocyanin O-methyltransferase. The transcription factors Anthocyanin1 (StAN1), basic Helix Loop Helix1 (StbHLH1), and StWD40 were more strongly expressed in red and purple potatoes. Expression of 12 other transcription factors was not associated with phenylpropanoid content, except for StMYB12B, which showed a negative relationship. Increased expression of AN1, bHLH1, and WD40 was also associated with environmentally mediated increases in tuber phenylpropanoids. Treatment of potato plantlets with sucrose induced hydroxycinnamic acids, flavonols, anthocyanins, structural genes, AN1, bHLH1, WD40, and genes encoding the sucrose-hydrolysing enzymes SUSY1, SUSY4, and INV2. Transient expression of StAN1 in tobacco leaves induced bHLH1, structural genes, SUSY1, SUSY4, and INV1, and increased phenylpropanoid amounts. StAN1 infiltration into tobacco leaves decreased sucrose and glucose concentrations. In silico promoter analysis revealed the presence of MYB and bHLH regulatory elements on sucrolytic gene promoters and sucrose-responsive elements on the AN1 promoter. These findings reveal an interesting dynamic between AN1, sucrose, and sucrose metabolic genes in modulating potato phenylpropanoids.

Accumulation of anthocyanins in grape berries is influenced by environmental factors (such as temperature and light) and supply of nutrients, i.e., fluxes of carbon and nitrogen feeding the berry cells. It is established that low nitrogen supply stimulates anthocyanin production in berry skin cells of red varieties. The present works aims to gain a better understanding of the molecular mechanisms involved in the response of anthocyanin accumulation to nitrogen supply in berries from field grown-plants. To this end, we developed an integrated approach combining monitoring of plant nitrogen status, metabolite measurements and transcript analysis. Grapevines (cv. Cabernet-Sauvignon) were cultivated in a vineyard with three nitrogen fertilization levels (0, 60 and 120 kg ha(-1) of nitrogen applied on the soil). Anthocyanin profiles were analyzed and compared with gene expression levels. Low nitrogen supply caused a significant increase in anthocyanin levels at two ripening stages (26 days post-véraison and maturity). Delphinidin and petunidin derivatives were the most affected compounds. Transcript levels of both structural and regulatory genes involved in anthocyanin synthesis confirmed the stimulation of the phenylpropanoid pathway. Genes encoding phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), flavonoid-3',5'-hydroxylase (F3'5'H), dihydroflavonol-4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX) exhibited higher transcript levels in berries from plant cultivated without nitrogen compared to the ones cultivated with 120 kg ha(-1) nitrogen fertilization. The results indicate that nitrogen controls a coordinated regulation of both positive (MYB transcription factors) and negative (LBD proteins) regulators of the flavonoid pathway in grapevine.

L-Phenylalanine ammonia-lyase (PAL) is the first enzyme in the biosynthesis of phenylpropanoid-derived plant compounds such as flavonoids, coumarins and the cell wall polymer lignin. The cell walls of grasses possess higher proportions of syringyl (S)-rich lignins and high levels of esterified coumaric acid compared with those of dicotyledonous plants, and PAL from grasses can also possess tyrosine ammonia-lyase (TAL) activity, the reason for which has remained unclear. Using phylogenetic, transcriptomic and in vitro biochemical analyses, we identified a single homotetrameric bifunctional ammonia-lyase (PTAL) among eight BdPAL enzymes in the model grass species Brachypodium distachyon. (13)C isotope labelling experiments along with BdPTAL1-downregulation in transgenic plants showed that the TAL activity of BdPTAL1 can provide nearly half of the total lignin deposited in Brachypodium, with a preference for S-lignin and wall-bound coumarate biosynthesis, indicating that PTAL function is linked to the characteristic features of grass cell walls. Furthermore, isotope dilution experiments suggest that the pathways to lignin from L-phenylalanine and L-tyrosine are distinct beyond the formation of 4-coumarate, supporting the organization of lignin synthesis enzymes in one or more metabolons.

To investigate the mechanisms whereby treatment with chitosan (CHN) is observed to increase the capacity of plants to resist pathogens, CHNs of different molecular weights (MWs) prepared by enzyme hydrolysis were used to treat rice cells in suspension culture and also rice seedlings. The results obtained with cultured cells showed that in this material CHN treatment could trigger a set of defence responses, including the production of hydrogen peroxide (H2O2), increases in the activities of phenylalanine ammonialyase (PAL; EC 4.3.1.5) and chitinase (CHI; EC 3.2.1.14), increases in transcription of defence-related genes beta-1,3-glucanase (glu) and chitinase (chi) and accumulation of pathogen-related protein (PR1). Furthermore, CHNs of different MWs were observed to have different capacities to induce defence responses. CHNs of low MWs were more effective at inducing the described defence responses than those of higher MWs. Enhanced defence against rice blast pathogen Magnaporthe grisea 97-23-2D1 was observed in rice seedlings treated with low MW CHNs compared to seedlings treated with higher MW CHNs. In all cases, suppressing the production of H2O2 by adding scavengers dimethylthiourea (DMTU), 2,5-dihydroxycinnamic acid methyl ester (DHC), catalase (Cat) or ascorbate (As) blocked the defence responses. These results indicate that CHNs of low MWs have a greater capacity to induce the production of H2O2, thus resulting in stronger defence responses, than those with higher MWs.

OBJECTIVE

To investigate the relationship between rural to urban migration and physical activity (PA) in India.

METHODS

6,447 (42% women) participants comprising 2077 rural, 2,094 migrants and 2,276 urban were recruited. Total activity (MET hr/day), activity intensity (min/day), PA Level (PAL) television viewing and sleeping (min/day) were estimated and associations with migrant status examined, adjusting for the sib-pair design, age, site, occupation, education, and socio-economic position (SEP).

RESULTS

Total activity was highest in rural men whereas migrant and urban men had broadly similar activity levels (p<0.001). Women showed similar patterns, but slightly lower levels of total activity. Sedentary behaviour and television viewing were lower in rural residents and similar in migrant and urban groups. Sleep duration was highest in the rural group and lowest in urban non-migrants. Migrant men had considerably lower odds of being in the highest quartile of total activity than rural men, a finding that persisted after adjustment for age, SEP and education (OR 0.53, 95% CI 0.37, 0.74). For women, odds ratios attenuated and associations were removed after adjusting for age, SEP and education.

CONCLUSIONS

Our findings suggest that migrants have already acquired PA levels that closely resemble long-term urban residents. Effective public health interventions to increase PA are needed.

BACKGROUND

Recently published results of quality of life (QoL) studies indicated different outcomes of palliative radiotherapy for brain metastases. This prospective multi-center QoL study of patients with brain metastases was designed to investigate which QoL domains improve or worsen after palliative radiotherapy and which might provide prognostic information.

METHODS

From 01/2007-01/2009, n=151 patients with previously untreated brain metastases were recruited at 14 centers in Germany and Austria. Most patients (82 %) received whole-brain radiotherapy. QoL was measured with the EORTC-QLQ-C15-PAL and brain module BN20 before the start of radiotherapy and after 3 months.

RESULTS

At 3 months, 88/142 (62 %) survived. Nine patients were not able to be followed up. 62 patients (70.5 % of 3-month survivors) completed the second set of questionnaires. Three months after the start of radiotherapy QoL deteriorated significantly in the areas of global QoL, physical function, fatigue, nausea, pain, appetite loss, hair loss, drowsiness, motor dysfunction, communication deficit and weakness of legs. Although the use of corticosteroid at 3 months could be reduced compared to pre-treatment (63 % vs. 37 %), the score for headaches remained stable. Initial QoL at the start of treatment was better in those alive than in those deceased at 3 months, significantly for physical function, motor dysfunction and the symptom scales fatigue, pain, appetite loss and weakness of legs. In a multivariate model, lower Karnofsky performance score, higher age and higher pain ratings before radiotherapy were prognostic of 3-month survival.

CONCLUSIONS

Moderate deterioration in several QoL domains was predominantly observed three months after start of palliative radiotherapy for brain metastases. Future studies will need to address the individual subjective benefit or burden from such treatment. Baseline QoL scores before palliative radiotherapy for brain metastases may contain prognostic information.

An ethylene response-related factor, GbERF1-like, from Gossypium barbadense cv. '7124' involved in the defence response to Verticillium dahliae was characterized. GbERF1-like transcripts present ubiquitously in various tissues, with higher accumulation in flower organs. GbERF1-like was also responsive to defence-related phytohormones and V. dahliae infection. The downregulation of GbERF1-like increased the susceptibility of cotton plants to V. dahliae infection, while overexpression of this gene improved disease resistance in both cotton and Arabidopsis, coupled with activation of the pathogenesis-related proteins. Further analysis revealed that genes involved in lignin synthesis, such as <em>PAL</em>, C4H, C3H, HCT, CCoAOMT, CCR and F5H, showed higher expression levels in the overexpressing cotton and Arabidopsis lines and lower expression levels in the RNAi cotton lines. The expression levels of these genes increased obviously when the GbERF1-like-overexpressing plants were inoculated with V. dahliae. Meanwhile, significant differences in the content of whole lignin could be found in the stems of transgenic and wild-type plants after inoculation with V. dahliae, as revealed by metabolic and histochemical analysis. More lignin could be detected in GbERF1-like-overexpressing cotton and Arabidopsis but less in GbERF1-like-silencing cotton compared with wild-type plants. The ratio of S and G monomers in GbERF1-like-overexpressing cotton and Arabidopsis increased significantly after infection by V. dahliae. Moreover, our results showed that the promoters of GhHCT1 and At<em>PAL</em>3 could be transactivated by GbERF1-like in vivo based on yeast one-hybrid assays and dual-luciferase reporter assays. Knockdown of GhHCT1 in GbERF1-like over-expressing cotton decreases resistance to V. dahliae. Collectively, our results suggest that GbERF1-like acts as a positive regulator in lignin synthesis and contributes substantially to resistance to V. dahliae in plants.

Herbaceous peony (Paeonia lactiflora Pall.) is an important ornamental plant which contains different flower colors. In this paper, eight genes encoding phenylalanine ammonialyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3'-hydroxylase (F3'H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), UDP-glucose: flavonoid 3-o-glucosyltransferase (UF3GT) were isolated. Moreover, the expression patterns of these eight genes and UF5GT in the flowers were investigated in three cultivars, that is, 'Hongyanzhenghui', 'Yulouhongxing' and 'Huangjinlun' with purplish-red, white and yellow flower respectively. Furthermore, flavonoid accumulation in the flowers was also analyzed. The results showed that in different organs, most of genes expressed higher in flowers than in other organs. During the development of flowers, all genes could be divided into four groups. The first group (PlPAL) was highly expressed in S1 and S4. The second group (PlCHS and PlCHI) was at a high expression level throughout the whole developmental stages. The third group (PlF3H, PlF3'H, PlDFR, PlANS and PlUF5GT) gradually decreased with the development of flowers. The fourth group (PlUF3GT) gradually increased during the flower development. In addition, anthoxanthins and anthocyanins were detected in 'Hongyanzhenghui' and 'Yulouhongxing', chalcones and anthoxanthins were found in 'Huangjinlun'. When different color flowers were concerned, low expression level of PlCHI induced most of the substrate accumulation in the form of chalcones and displaying yellow, changing a small part of substrates to anthoxanthins, and there was no anthocyanin synthesis in 'Huangjinlun' because of low expression level of DFR. In 'Yulouhongxing', massive expressions of upstream genes and low expression of DFR caused synthesis of a great deal of anthoxanthins and a small amount of colorless anthocyanins. In 'Hongyanzhenghui', a large number of colored anthocyanins were changed from anthoxanthins because of PlDFR, PlANS and PlUF3GT high expressions. These results would provide us a theoretical basis to understand the formation of P. lactiflora flower colors.

Salicylic acid (SA) is an important plant hormone that is best known for mediating host responses upon pathogen infection. Its role in plant defense activation is well established, but its biosynthesis in plants is not fully understood. SA is considered to be derived from two possible pathways; the ICS and PAL pathway, both starting from chorismate. The importance of both pathways for biosynthesis differs between plant species, rendering it hard to make generalizations about SA production that cover the entire plant kingdom. Yet, understanding SA biosynthesis is important to gain insight into how plant pathogen responses function and how pathogens can interfere with them. In this review, we have taken a closer look at how SA is synthesized and the importance of both biosynthesis pathways in different plant species.

To understand human papillomavirus type 16 (HPV-16) gene regulation, it is necessary to understand HPV-16 RNA processing. HPV-16 encodes multiple 5'- and 3'-splice sites and two polyadenylation signals pAE and pAL (Figure 1). The major 3'-splice site on the HPV-16 genome (SA3358) is used for generation of E6, E7, E4, L1 and L2 mRNAs. It encodes a suboptimal splice signal but is under control of a strong enhancer that renders SA3358 one of the most efficiently used splice sites on the HPV-16 genome. Thereby SA3358 indirectly blocks HPV-16 late gene expression. The early polyA signal is also under control of the early UTR sequence and multiple RNA elements in the L2 coding region that interact with hnRNP H. The two splice sites SD3632 and SA5639 are used exclusively by late mRNAs and are under control of multiple splicing silencer elements. The silencers at SA5639 are located in the L1 coding region and interact with hnRNP A1. So far, only polypyrimidine tract binding protein (PTB) has been shown to induce late gene expression.

A method is described in which palmitate-conjugated protein A (pal-protein A) incorporated onto cell membranes is used to anchor antibodies on the surface of cells. Pal-protein A was stabilized on the cell surfaces via insertion of the hydrophobic palmitate moieties into the phospholipid bilayer of the plasma membrane. This membrane-associated pal-protein A retained its binding affinity for the Fc region of rabbit immunoglobulin G (IgG) molecules. Using this system, cells can be coated with antibody (Ab) molecules of a desired specificity that can serve as artificial receptors for soluble or cell surface antigens (Ags). In this report, we show that A22.E10 T hybridoma cells coated with rabbit anti-mouse IgG (RaMIgG) exhibited increased adhesiveness to surface IgG positive mouse B cells. A high level of heterotypic intercellular conjugation was observed in cells coated with specific artificial receptors as compared to cells coated with nonspecific artificial receptors.

Photoaffinity labeling (PAL) using a chemical probe to covalently bind its target in response to activation by light has become a frequently used tool in drug discovery for identifying new drug targets and molecular interactions, and for probing the location and structure of binding sites. Methods to identify the specific target proteins of hit molecules from phenotypic screens are highly valuable in early drug discovery. In this review, we summarize the principles of PAL including probe design and experimental techniques for in vitro and live cell investigations. We emphasize the need to optimize and validate probes and highlight examples of the successful application of PAL across multiple disease areas.

OBJECTIVE

The objectives of this study were to investigate whether a classification of triticale genotypes into drought-tolerant and drought-sensitive types based on field performance trials correlates with a classification based on measurements of some physiological and biochemical parameters in greenhouse conditions. In addition, an examination was carried out of whether ferulic acid, as the main origin of the blue fluorescence produced, contributes to drought tolerance.

METHODS

Ten winter triticale genotypes were examined, five known to be drought tolerant and five drought sensitive. Measurements of the osmotic potential, leaf gas exchange, chlorophyll fluorescence, and blue and red fluorescence were performed. In addition, analysis of the total pool of phenolic compounds and ferulic acid as well as the measurements of PAL (l-phenylalanine ammonia-lyase) activity were carried out.

RESULTS

In agreement with field trials, three out of five cultivars ('Lamberto', 'Timbo' and 'Piano') were classified as drought tolerant. However, in the case of cultivar 'Babor', included in the group of drought-sensitive cultivars, the values obtained for some measured parameters were close to (F(v)(')/F(m)('), phenolics content, osmotic potential) or even better than (non-photochemical quenching, red and blue fluorescence, ferulic acid content) those for drought-tolerant genotypes. Cultivars 'Imperial', 'Ticino', 'Trimaran' and 'Boreas' were included in the drought-sensitive group, whereas cultivars 'Focus' and 'Kitaro' were included in the moderately sensitive group.

CONCLUSIONS

The experiments confirmed that the period of flowering, the critical phase for plants as far as water demand is concerned, is suitable for plant screening and differentiation due to their tolerance to drought. The most important criteria which enabled creation of the ranking list of plants, from those sensitive to drought to those tolerant to drought, were the ability to perform the process of osmoregulation, the efficiency of the utilization of excitation energy by the photosynthetic apparatus and the functioning of protective mechanisms involving the level of ferulic acid in leaf tissues.

The occurrence of atrial fibrillation (AF), especially in patients with mitral regurgitation (MR), is related to the degree of left atrial (LA) myopathy, remodeling and fibrosis, that are responsible of LA electrical inhomogeneity and abnormal conduction velocities. Speckle tracking echocardiography (STE) has recently enabled the quantification of longitudinal myocardial LA deformation dynamics. Our aim was to investigate by STE the effects of the occurrence of paroxysmal AF on LA myocardial deformation, in a population of patients with asymptomatic chronic MR. We compared two groups of a total of 197 patients with MR: 54 with history of paroxysmal AF and 143 with MR alone. Subgroups were created according to MR degree. Peak atrial longitudinal strain (PALS) was measured in all subjects. Values were obtained by averaging all segments (global PALS), measured in the 4-chamber and 2-chamber views. Compared to the mild MR group (46.1 ± 4.9%), global PALS was lower in moderate MR group (22.1 ± 5.8%) and further reduced in the severe MR group (13.9 ± 4.2%; overall P < 0.0001 by ANOVA, P < 0.05 for all pair-wise comparisons). Besides, in each MR group, patients with history of paroxysmal AF presented a global PALS significantly reduced (overall P < 0.0001 by ANOVA). After multivariate analysis, global PALS was significantly and independently associated with paroxysmal AF. STE enables noninvasive quantification of LA dysfunction due to MR and paroxysmal AF. MR have a major negative impact on LA function. In patients with MR, the history of paroxysmal AF is associated to a further impair of LA myocardial reservoir function.

The immunophenotype of spindle cells in epidemic, endemic, and classic (sporadic) Kaposi's sarcoma (KS) lesions was defined by the demonstration of various cell markers and compared with that of KS-derived cell lines. No significant histological or immunophenotypic differences were observed between the three clinical types of KS at comparable stages. The spindle-cell compartment of the different KS types was composed predominantly of a mixture of proliferating CD45+/CD68+ bone-marrow-derived monocytes and TE7+/collagen+ fibroblastic cells with varying expression of EN4/PAL-E/CD31/CD34/CD36 endothelial-associated antigens and/or smooth-muscle-specific alpha-actin (alpha-actin). The latter cells appeared to represent transitional forms of fibroendothelial and fibromyocytic cells. The in vitro cultured KS-derived cell lines (KS-3, KS-6, and KS-8) expressed the fibroblastic antigen TE7 and smooth-muscle-specific alpha-actin but not leukocytic or endothelial-associated antigens consistent with the phenotype of fibromyoid spindle cells of primary lesions. Neither HIV antigen nor provirus DNA was demonstrable in the epidemic KS lesions. The observed heterogeneity of the spindle-cell compartment further substantiates the view that Kaposi's sarcoma, irrespective of clinical setting, expresses salient features more compatible with reactive, tumor-like lesion than clonal sarcoma.

We cloned and expressed in Escherichia coli a gene encoding an 18-kDa outer membrane protein (Omp18) from Campylobacter jejuni ATCC 29428. The nucleotide sequence of the gene encoding Omp18 was determined, and an open reading frame of 165 amino acids was revealed. The amino acid sequence had the typical features of a leader sequence and a signal peptidase II cleavage site at the N-terminal part of Omp18. Moreover, the sequence had a high degree of similarity to the peptidoglycan-associated outer membrane lipoprotein P6 of Haemophilus influenzae and the peptidoglycan-associated lipoprotein PAL of E. coli. Southern blot analysis in which the cloned gene was used as a probe revealed genes similar to that encoding Omp18 in all species of the thermophilic group of campylobacters as well as Campylobacter sputorum. All campylobacters tested expressed a protein with a molecular mass identical to that of Omp18. The protein reacted immunologically with polyclonal antibodies directed against Omp18 from C. jejuni. PCR amplification of the gene encoding Omp18 with specific primers and subsequent restriction enzyme analysis of the amplified DNA fragments showed that the gene for Omp18 is highly conserved in C. jejuni strains isolated from humans, dogs, cats, calves, and chickens but is different in other Campylobacter species. In order to obtain pure recombinant Omp18 protein for serological assays, the cloned gene for Omp18 was genetically modified by replacing the signal sequence with a DNA segment encoding six adjacent histidine residues. Expression of this construct in E. coli allowed purification of the modified protein (Omp18-6xHis) by metal chelation chromatography. Sera from patients with past C. jejuni infection reacted positively with Omp18-6xHis, while sera from healthy blood donors showed no reaction with this antigen. Omp18, which is an outer membrane protein belonging to the family of PALs is well conserved in C. jejuni and is highly immunogenic. It is therefore a good candidate as an antigen for the serological diagnosis of past C. jejuni infections.

OBJECTIVE

The relative influence of dietary factors vs physical activity on cardiovascular risk factors are poorly understood. We investigated these factors in a population whose traditional diet may have both positive (high plant-based) and negative (high refined carbohydrate) aspects, and whose physical activity levels (PALs) vary widely.

METHODS

Cross-sectional study.

METHODS

A total of 130 weight stable adults aged 35-49 y (BMI 18-35 kg/m(2)) living in urban Beijing, China.

METHODS

Dietary intake (by food frequency questionnaire), PAL as the ratio of predicted total to resting energy expenditure), percent body fat (by deuterium oxide dilution), and central adiposity (waist circumference and waist to hip ratio) were assessed. Biochemical parameters (total cholesterol, low- and high-density lipoprotein cholesterol (LDL-C; HDL-C), triglyceride (TG), apolipoproteins A-I and B, glucose, insulin, and homocysteine and its related vitamins), blood pressure and presence of the metabolic syndrome (having>>/=3 risk factors of central adiposity, HDL-C, TG, glucose, blood pressure) were also examined.

RESULTS

Mean values for cardiovascular risk factors were relatively low, but 19% of subjects had the metabolic syndrome. Using validated methods for measuring food intake and energy expenditure, we found that an adverse cardiovascular risk profile was associated with a diet high in carbohydrate, low in polyunsaturated fat, and low in fruit and vegetables, independent of body fatness and its distribution. While dietary factors predicted individual cardiovascular risk factors more consistently than PAL, avoidance of low PAL reduced the risk of having the metabolic syndrome.

CONCLUSIONS

These results suggest that, regardless of total body fatness and fat distribution, multiple unfavorable dietary factors and low physical activity independently increase the risk for cardiovascular disease. Avoidance of a sedentary lifestyle additionally reduces the risk of developing the metabolic syndrome.

Shc adaptor proteins play a role in linking activated cell surface receptors to the Ras signaling pathway in response to receptor mediated tyrosine kinase activation. While the function of Shc in the activation of the Ras pathway via binding to Grb2 has been well characterized, it is becoming increasingly apparent that Shc participates in additional signaling pathways through interactions with other cytoplasmic proteins. Using the yeast two-hybrid system, we have identified a unique Shc binding protein designated PAL (Protein expressed in Activated Lymphocytes) with no similarity to other known proteins. mPAL binds specifically to the Shc SH2 domain and unlike previously described Shc SH2 domain-protein interactions, the association of mPAL and Shc is phosphotyrosine-independent. Both mPAL RNA and protein expression are restricted to tissues containing actively dividing cells and proliferating cells in culture. mPAL expression is induced upon growth factor stimulation and is down-regulated upon growth inhibition. This pattern, and timing of mPAL expression and its association with the Shc adaptor molecule suggests a role for this protein in signaling pathways governing cell cycle progression.

During immune responses the initial activation of B cells takes place in T cell zones of periarteriolar lymphoid sheaths (PALS) of the splenic white pulp. After initial activation, B cells migrate into the primary follicles and, in association with follicular dendritic cells (FDCs), undergo clonal expansion and differentiation giving rise to germinal centers (GCs). Peanut agglutinin binding (PNA+) cells of the GC differentiate further into memory or plasma cells. Here we report that in tumor necrosis factor receptor 1-deficient mice (TNFR1(-/-)), the location of B cells was altered and that plasma cells were abnormally distributed in the splenic PALS. In contrast to lymphotoxin alpha-deficient mice (LTalpha-/-), bone marrow or fetal liver transplantation did not correct the abnormal organization of the spleen, location of B cells, the lack of an FDC network, nor the antibody response in TNFR1(-/-) mice. These results argue for a crucial role of TNFR1 expression on nonhematopoietic cells for the maintenance of the splenic architecture and proper B cell location. In addition, the lack in development of an FDC network after adoptive transfer suggests that either FDCs are not of bone marrow origin or that they depend on signals from nonhematopoietic cells for maturation.