Inflammatory bowel disease (IBD), majorly include Crohn's disease (CD) and ulcerative colitis (UC), is chronic and relapsing inflammatory disorders of the gastrointestinal tract, which treatment options remain limited. Here we examined the therapeutic effects of an isoquinoline alkaloid, Palmatine (Pal), on mice experimental colitis induced by dextran sulfate sodium (DSS) and explored underlying mechanisms. Colitis was induced in BALB/c mice by administering 3% DSS in drinking water for 7 days. Pal (50 and 100 mg kg-1) and the positive drug Sulfasalazine (SASP, 200 mg kg-1) were orally administered for 7 days. Disease activity index (DAI) was evaluated on day 8, and colonic tissues were collected for biochemistry analysis. The fecal microbiota was characterized by high-throughput Illumina MiSeq sequencing. And plasma metabolic changes were detected by UPLC-MS. Our results showed that Pal treatment significantly reduced DAI scores and ameliorated colonic injury in mice with DSS-induced colitis. Mucosal integrity was improved and cell apoptosis was inhibited. Moreover, gut microbiota analysis showed that mice received Pal-treatment have higher relative abundance of Bacteroidetes and Firmicutes, but reduced amount of Proteobacteria. Moreover, Pal not only suppressed tryptophan catabolism in plasma, but also decreased the protein expression of indoleamine 2,3-dioxygenase 1 (IDO-1, the rate-limiting enzyme of tryptophan catabolism) in colon tissue. This is consolidated by molecular docking, which suggested that Pal is a potent IDO-1 inhibitor. Taken together, our findings demonstrate that Pal ameliorated DSS-induced colitis by mitigating colonic injury, preventing gut microbiota dysbiosis, and regulating tryptophan catabolism, which indicated that Pal has great therapeutic potential for colitis.

A prominent 19 kDa surface antigen of Legionella pneumophila, cloned in Escherichia coli, was found to be intimately associated with peptidoglycan. The DNA region encoding this antigen was mapped on an 11.9 kb plasmid by means of deletion analysis and transposon mutagenesis. PhoA+ gene fusions, gene-rated by TnphoA insertions into this region, confirmed the presence of a gene encoding a secreted protein. PhoA+ transposon insertions were also associated with loss of the 19 kDa antigen in immunoassays using a monoclonal antibody (mAb1E9) and the replacement of the 19 kDa antigen with larger fusion proteins in immunoblots using Legionella immune serum. A 1540bp PstI fragment carrying the gene was sequenced, and the open reading frame encoding the antigen was identified. The gene encodes a polypeptide 176 amino acid residues long and 18913Da in size. The presence of a signal sequence of 22 amino acids with a consensus sequence for cleavage by signal peptidase II indicates that the antigen is a lipoprotein, and striking similarity with peptidoglycan-associated lipoproteins (PALs) from E. coli (51% amino acid homology) and Haemophilus influenzae (55% homology) is noted. We conclude that the 19kDa antigen of L. pneumophila is the structural equivalent of the PAL found in other Gram-negative species and suggest that its post-translational acylation may explain its potency as an immunogen.

BACKGROUND

Tea is the most popular nonalcoholic beverage worldwide for its pleasant characteristics and healthful properties. Catechins, theanine and caffeine are the major natural products in tea buds and leaves that determine tea qualities such as infusion colors, tastes and fragrances, as well as their health benefits. Shading is a traditional and effective practice to modify natural product accumulation and to enhance the tea quality in tea plantation. However, the mechanism underlying the shading effects is not fully understood. This study aims to explore the regulation of flavonoid biosynthesis in Camellia sinensis under shading by using both metabolomic and transcriptional analyses.

RESULTS

While shading enhanced chlorophyll accumulation, major catechins, including C, EC, GC and EGC, decreased significantly in tea buds throughout the whole shading period. The reduction of catechins and flavonols were consistent with the simultaneous down-regulation of biosynthetic genes and TFs associated with flavonoid biosynthesis. Of 16 genes involved in the flavonoid biosynthetic pathway, F3'H and FLS significantly decreased throughout shading while the others (PAL, CHSs, DFR, ANS, ANR and LAR, etc.) temporally decreased in early or late shading stages. Gene co-expression cluster analysis suggested that a number of photoreceptors and potential genes involved in UV-B signal transductions (UVR8_L, HY5, COP1 and RUP1/2) showed decreasing expression patterns consistent with structural genes (F3'H, FLS, ANS, ANR, LAR, DFR and CHSs) and potential TFs (MYB4, MYB12, MYB14 and MYB111) involved in flavonoid biosynthesis, when compared with genes in the UV-A/blue and red/far-red light signal transductions. The KEGG enrichment and matrix correlation analyses also attributed the regulation of catechin biosynthesis to the UVR8-mediated signal transduction pathway. Further UV-B treatment in the controlled environment confirmed UV-B induction on flavonols and EGCG accumulation in tea leaves.

CONCLUSIONS

We proposed that catechin biosynthesis in C. sinensis leaves is predominantly regulated by UV through the UVR8-mediated signal transduction pathway to MYB12/MYB4 downstream effectors, to modulate flavonoid accumulation. Our study provides new insights into our understanding of regulatory mechanisms for shading-enhanced tea quality.

Phenylalanine ammonia-lyase is the first enzyme of general phenylpropanoid pathway. A PAL gene, designated as BoPALPALPALPAL gene found in angiosperm plant. Several putative cis-acting elements such as P box, GT-1motif, and SOLIPs involved in light responsiveness were found in the 5'-flanking sequence of BoPALPALPALPALPAL was about 80 kDa, indicating that BoPALPALPALs reported in other plants.

The objective of this study is to estimate the direct medical cost of end-of-life and palliative (EOL/PAL) care for cancer patients during the last six months of their lives--or, during the period from diagnosis to death, if briefer--in 2002 and 2003, in Ontario, Canada. A linkage of cancer registry and administrative data is used to determine the costs of health care resources used during the EOL/PAL care period. Costs are analyzed by cancer diagnosis, location of death, and type of service. The total Ontario Ministry of Health-funded cost of EOL/PAL care for cancer patients is estimated to be about CAD$544 million per year, with an average per patient cost of about $25,000 in 2002-2003. Our results suggest that acute care consumes 75 percent of EOL/PAL funding and that only a small proportion of health care services used by EOL/PAL care cancer patients is likely to be formal palliative care.

Defense responses to inoculation with Fusarium oxysporum SCHLECHT f. sp. lupini were studied in embryo axes of Lupinus luteus L. cv. Polo cultured on a medium with sucrose (60 mM) or without it. Exogenous sucrose caused a marked endogenous increase in concentrations of sucrose, glucose and fructose in embryo axes. In axes cultured with sucrose, high performance liquid chromatography (HPLC) revealed generally higher levels of isoflavone glycosides (particularly until 48 h of culture) and free aglycones (genistein, wighteone, luteone). Inoculation resulted in a considerable decline in soluble carbohydrates between 24 and 72 h of culture. Simultaneously, the infection stimulated an increase in the level of free isoflavone aglycones in inoculated embryo axes, as compared to non-inoculated ones. Concentrations of free aglycones (i.e. genistein, wighteone and luteone) after infection were particularly high in inoculated embryo axes fed with sucrose. Genistein was a better inhibitor to F. oxysporum growth than genistein 7-O-glucoside tested. Exogenous sucrose also stimulated the activity of phenylalanine ammonialyase (PAL, EC 4.3.1.5)--an important enzyme initiating phenylpropanoid metabolism. After infection of tissues, a strong increase was observed in the activity of PAL and beta-glucosidase (EC 3.2.1.21)--an enzyme hydrolyzing isoflavone glycosides. Furthermore, the growth of inoculated embryo axes cultured with sucrose was less inhibited as a result of infection than inoculated axes cultured under carbohydrate deficiency conditions. Additionally, it had been reported previously that disease symptoms of embryo axes growing in the presence of sucrose were less intensive [30]. These results suggest that soluble sugars are involved in the mechanism of resistance, as they can stimulate phenylpropanoid metabolism and contribute to the increase in concentration of isoflavonoids, which are important elements of the defense system of legumes.

OBJECTIVE

To assess the reliability and validity of scoring instruments designed to measure clinical performance during simulated resuscitations requiring the use of Pediatric Advanced Life Support (PALS) algorithms.

METHODS

Pediatric residents were invited to participate in an educational trial involving simulated resuscitations that employ PALS algorithms. Each subject participated in a session comprised of four scenarios (asystole, dysrhythmia, respiratory arrest, shock). Video-recorded sessions were independently reviewed and scored by four raters using instruments designed to measure performance in terms of timing, sequence, and quality. Validity was assessed by two-factor analysis of variance with postgraduate year (PGY-1 versus PGY-2) as an independent variable. Reliability was assessed by calculation of overall interrater reliability (IRR) as well as a generalizability study to estimate variance components of individual measurement facets (scenarios, raters) and associated interactions.

RESULTS

20 subjects were scored by four raters. Based on a two-factor ANOVA, PGY-2s outperformed PGY-1s (p<0.05); significant differences in difficulty existed between the four scenarios, with dysrhythmia scores being the lowest. Overall IRR was high (0.81) and most variance could be attributed to subject (17%), scenario (13%), and the interaction between subject and scenario (52%); variance attributable to rater was minimal (1.4%).

CONCLUSIONS

The instruments assessed in this study measure clinical performance during PALS scenarios in a reliable and valid manner. Measurement error could be minimized further through the use of additional scenarios but additional raters, for a given scenario, would not improve reliability. Further studies should assess validity of measurement with respect to actual clinical performance during resuscitations.

Phenylalanine ammonia-lyase (PAL) is a key enzyme in the synthesis of phenolic compounds, which play a prominent role in graft union formation, including the marked effects of their accumulation in incompatibility response. The purpose of this study was to assess changes in the abundance of PAL mRNA during graft union development. Partial cDNA clones encoding the enzyme were isolated from in vitro callus tissue in the apricot (Prunus armeniaca L.) cultivar Moniqui and the plum rootstock Marianna 2624 (Prunus munsoniana x Prunus cerasifera). The deduced partial amino acid sequence showed high homology with PAL genes from other plant species. We monitored PAL expression 5, 10, 15 and 20 days after the establishment of in vitro callus unions. The levels of PAL mRNA increased 5 days after grafting in both compatible and incompatible unions. Nevertheless, significant differences were observed at the transcript level through both types of combinations from the second week. The results showed a higher level of PAL transcription in graft unions of incompatible partners, where a lack of adaptation between stock and scion takes place. The level of scion-stock compatibility was related to the PAL expression pattern. In addition, cell walls of the callus cells were not stained by phloroglucinol-HCl, indicating that the increased PAL expression did not result in the formation of lignin. However, staining with Naturstoff reagent A confirmed the highest accumulation of soluble and wall-bound phenolic compounds at the graft interface of incompatible unions.

Pal amidase, encoded by pneumococcal bacteriophage Dp-1, represents one step beyond in the modular evolution of pneumococcal murein hydrolases. It exhibits the choline-binding module attaching pneumococcal lysins to the cell wall, but the catalytic module is different from those present in the amidases coded by the host or other pneumococcal phages. Pal is also an effective antimicrobial agent against Streptococcus pneumoniae that may constitute an alternative to antibiotic prophylaxis. The structural implications of Pal singular structure and their effect on the choline-amidase interactions have been examined by means of several techniques. Pal stability is maximum around pH 8.0 (Tm approximately 50.2 degrees C; DeltaHt = 183 +/- 4 kcal mol(-1)), and its constituting modules fold as two tight interacting cooperative units whose denaturation merges into a single process in the free amidase but may proceed as two well resolved events in the choline-bound state. Choline titration curves reflect low energy ligand-protein interactions and are compatible with two sets of sites. Choline binding strongly stabilizes the cell wall binding module, and the conformational stabilization is transmitted to the catalytic region. Moreover, the high proportion of aggregates formed by the unbound amidase together with choline preferential interaction with Pal dimers suggest the existence of marginally stable regions that would become stabilized through choline-protein interactions without significantly modifying Pal secondary structure. This structural rearrangement may underlie in vitro "conversion" of Pal from the low to the full activity form triggered by choline. The Pal catalytic module secondary structure could denote folding conservation within pneumococcal lytic amidases, but the number of functional choline binding sites is reduced (2-3 sites per monomer) when compared with pneumococcal LytA amidase (4-5 sites per monomer) and displays different intermodular interactions.

Chlorogenic acid (CGA) is the major phenolic sink in potato tubers and can constitute over 90% of total phenylpropanoids. The regulation of CGA biosynthesis in potato and the role of the CGA biosynthetic gene hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase (HQT) was characterized. A sucrose induced accumulation of CGA correlated with the increased expression of phenylalanine ammonia-lyase (PAL) rather than HQT. Transient expression of the potato MYB transcription factor StAN1 (anthocyanin 1) in tobacco increased CGA. RNAi suppression of HQT resulted in over a 90% reduction in CGA and resulted in early flowering. The reduction in total phenolics and antioxidant capacity was less than the reduction in CGA, suggesting flux was rerouted into other phenylpropanoids. Network analysis showed distinct patterns in different organs, with anthocyanins and phenolic acids showing negative correlations in leaves and flowers and positive in tubers. Some flavonols increased in flowers, but not in leaves or tubers. Anthocyanins increased in flowers and showed a trend to increase in leaves, but not tubers. HQT suppression increased biosynthesis of caffeoyl polyamines, some of which are not previously reported in potato. Decreased PAL expression and enzyme activity was observed in HQT suppressed lines, suggesting the existence of a regulatory loop between CGA and PAL. Electrophysiology detected no effect of CGA suppression on potato psyllid feeding. Collectively, this research showed that CGA in potatoes is synthesized through HQT and HQT suppression altered phenotype and redirected phenylpropanoid flux.

OBJECTIVE

The aims of this study were to (1) compare prospectively the clinical and radiographic changes in periodontal and peri-implant conditions, (2) investigate the association of changes in periodontal parameters and peri-implant conditions over a mean observation period of 10 years (8-12 years) after implant installation, and (3) evaluate patient risk factors known to aggravate the periodontal conditions for their potential influence on the peri-implant tissue status.

METHODS

Eighty-nine partially edentulous patients with a mean age of 58.9 years (28-88 years) were examined at 1 and 10 years after implant placement. The patients contributed with 179 implants that were placed after comprehensive periodontal treatment and restored with crowns or fixed partial dentures. One hundred and seventy-nine matching control teeth were chosen as controls. Also, the remaining teeth (n=1770) in the dentitions were evaluated. Data on smoking habits and general health aspects were collected at 1 and 10 years as well.

RESULTS

At 10 years, statistically significant differences existed between implants and matching control teeth with regard to most of the clinical and radiographic parameters (P<0.01) with the exception of plaque index (PII) and recession. Multiple regression analyses were performed to associate combinations of periodontal diagnostic parameters to the peri-implant conditions: probing attachment level (PAL) at implants at 10 years was associated with implant location, full-mouth probing pocket depth (PPD) and full-mouth PAL (P=0.0001, r2=0.36). PPD at implants at 10 years correlated to implant location, full-mouth PPD and full-mouth PAL (P<0.001, r2=0.47). Marginal bone level at implants at 10 years was significantly associated to smoking, general health condition, implant location, full-mouth PAL and change over time in full-mouth PPD (P<0.001, r2=0.39).

CONCLUSIONS

These results present evidence for the association between periodontal and peri-implant conditions and the changes in these tissues over 10 years in partially edentulous patients.

BACKGROUND

The purpose of this study was to compare the accuracy of the Root ZX II Apex Locator (RZX), the Elements Apex Locator (ELE), and the Precision Apex Locator (PAL).

METHODS

Forty single-rooted extracted teeth were decoronated and the root canals coronally flared. Actual canal lengths were determined by inserting a #10 file until the tip was visualized (12.8x magnification) just within the apical foramina. Teeth were mounted in gelatin conducting medium and randomly tested with each electronic apex locator (EAL) to determine the electronic canal length. Differences between the electronic and actual canal lengths were calculated.

RESULTS

The mean differences were -0.02 mm, 0.13 mm, and 0.15 mm for the RZX, PAL, and ELE, respectively. One-way analysis of variance showed a highly significant difference among EALs (p = 0.003). Student-Newman-Keuls post hoc analysis found significant differences between the RZX and the PAL and between the RZX and the ELE at p < 0.05. No significant difference was noted between the PAL and the ELE. The proportion of electronic canal length measurements falling within +/-0.5 mm of the actual canal lengths for the EALs was as follows: 97.5% for the RZX, 95% for the PAL, and 90% for the ELE.

CONCLUSIONS

The RZX was the most accurate at locating the apical foramen compared with the ELE and the PAL.

OBJECTIVE

The effects of the recommended 30:2 compression:ventilation (C:V) ratio on chest compression rate (CR), compression depth (CD), compression pressure (CP) and rescuer fatigue is unknown during pediatric CPR. We hypothesized that a 30:2 C:V ratio will decrease compression depth and compression pressure and increase rescuer fatigue compared with a 15:2 ratio.

METHODS

Randomized crossover observational study.

METHODS

Adolescent, child and infant manikins were modified to digitally record compression rate, compression depth, compression pressure and total compression cycles (CC). BLS or PALS certified volunteers were randomized to five CPR groups: adolescent (AD), child 1-hand (OH), child 2-hand (TH), infant two-finger (TF) and infant two-thumb (TT). Each rescuer performed each ratio for 5 min with the order randomized. Rescuer heart rate (HR) and respiratory rate (RR) were recorded continuously during CPR and used to determine the recovery time (RT) for HR/RR to return to baseline. Data (mean+/-S.D.) were contrasted by paired differences for quantitative data and the sign rank test for ordinal data.

RESULTS

Eighty subjects (16 per group) were randomized. The peak compression pressure and compression rate were not different within any group, but total compression cycle were higher in all 30:2 groups. Compression depth (mm) was not significantly different within any group. The rescuer's HR (bpm) increased significantly during 30:2 CPR in AD and OH group with no significant differences in RR and recovery time. Subjects reported that 15:2 CPR was easier to perform (P<0.001).

CONCLUSIONS

During single rescuer pediatric BLS, more compression cycles were achieved with 30:2 C:V ratio without effect on compression depth, pressure and rate. Increased HR with 30:2 C:V ratio was noted during larger manikin CPR without subjective difference of reported fatigue. Most rescuers in AD and TF group did not achieve recommended compression depth regardless of C:V ratio.

Phenylketonuria (PKU) is a metabolic disorder due primarily to mutations in the PAH gene that impair both phenylalanine hydroxylase activity and disposal of l-phenylalanine from the normal diet. Excess phenylalanine is toxic to cognitive development and a low-phenylalanine diet prevents mental retardation, but it is a difficult therapeutic option. Previous studies with recombinant phenylalanine ammonia-lyase, PAL, demonstrated pharmacologic and physiologic proofs of principle for PAL as an alternative therapy for PKU but its immunogenicity was problematic. From a series of formulations of linear and branched polyethylene glycols chemically conjugated to PAL, we have created a parenteral therapeutic agent for PKU treatment. All the pegylated molecules were fully characterized in vitro and the most promising formulations were then tested in vivo in the PKU mouse model. The linear 20-kDa PEG-PAL combination abolished in vivo immunogenicity after repeated challenge while retaining full catabolic activity against phenylalanine, suggesting potential as a novel PKU therapeutic.

The secretion of proteolytic enzymes by dermatophytes is a key factor in their invasion and subsequent dissemination through the stratum corneum of the host. During the first stages of infection, dermatophytes respond to the skin by de-repressing a number of genes coding for proteins and enzymes such as adhesins, lipases, phosphatases, DNAses, non-specific proteases, and keratinases. These proteins have their optimal activity at acidic pH values, which matches the acidic pH of human skin, allowing the pathogen to adhere and penetrate the host tissue, scavenge nutrients and overcome host defence mechanisms. The conserved PacC/Rim101p signal transduction pathway mediates diverse metabolic events involved in ambient pH sensing and in the virulence of pathogenic microorganisms. The seven dermatophyte genomes analysed here revealed the presence of the PacC/Rim101p pH-responsive signal transduction pathway, which consists of the six pal genes (palA, B, C, F, H and I) and the transcription factor PacC. The PacC binding site was present in the promoter regions of pacC, palB, palI and palH genes of all dermatophytes, suggesting functional equivalency with the signalling cascade of other fungi. Moreover, the promoter region of pacC gene of the seven dermatophytes had multiple PacC DNA-binding sites, suggesting that these genes, like their homologues in model fungi, are auto-regulated.

Target identification of biologically active molecules such as natural products, synthetic small molecules, peptides, and oligonucleotides mainly relies on affinity chromatography, activity-based probes, or photoaffinity labeling (PAL). Amongst them, activity-based probes and PAL have offered great advantages in target identification technology due to their ability to form covalent bonds with the corresponding targets. Activity-based probe technology mainly relies on the chemical reactivity of the target proteins, thereby limiting the majority of the biological targets to enzymes or proteins which display reactive residues at the probe-binding site. In general, the probes should bear a reactive moiety such as an epoxide, a Michael acceptor, or a reactive alkyl halide in their structures. On the other hand, photoaffinity probes (PAPs) are composed of a target-specific ligand and a photoactivatable functional group. When bound to the corresponding target proteins and activated with wavelength-specific light, PAPs generate highly reactive chemical species that covalently cross-link proximal amino acid residues. This process is better known as PAL and is widely employed to identify cellular targets of biologically active molecules. This review highlights recent advances in target identification by PAL, with a focus on the structure and chemistry of the photoaffinity probes developed in the recent decade, coupled to the target proteins identified using these probes.

The purpose of the present study was to assess different aspects of physical activity and fitness in order to develop a basis for sport programmes for overweight and obese children. Eighty-eight prepubertal children (49 boys, 39 girls, 4.8-11.4 years old, 61% obese, 14% overweight and 25% normal weight) were examined. Body composition was assessed by combined use of anthropometrics and bioelectrical impedance analysis. Resting energy expenditure (REE) and total energy expenditure (TEE) were measured by indirect calorimetry (IC) and individually calibrated 24-h heart rate (HR) monitoring, respectively. Activity-related energy expenditure (AEE) and physical activity level (PAL) were calculated from TEE and REE. Fitness [assessed by O2-pulse, respiratory exchange ratio (RER) at submaximal work intensities] was determined by ergometry. The maximal isometric muscle strength of the legs (m. quadriceps, Fa max, m. ischiocruralis, Fb max) was measured by computer tensiometry. Children were grouped according to their nutritional state, AEE, O2-pulse and muscle strength. When compared with normal weight children, obese and overweight children had increased fat mass (FM), fat-free mass (FFM), waist-to-hip ratio and REE, but no group differences were observed for TEE, AEE, and PAL. Obese and overweight children spent more hours per day watching TV. After correction for body weight and FFM, no group differences in REE were observed, but normal weight children had a higher O2-pulse than overweight and obese children. By contrast, RER was increased in the latter group. The fittest group had the lowest body weight, BMI, FM and FFM. Children with a low O2-pulse spent more hours per day watching TV. Grouping children according to their degree of muscle strength, younger children (4-7.5 years) did not show group differences in nutritional state, energy expenditure, physical activity and fitness. However, in the group of 7.6- to 11-year-old children, those with the greatest muscle strength and FFM had reduced BMI, skin folds, FM and FFM. FM correlated inversely with O2-pulse, but was not associated with TEE, AEE, PAL or muscle strength. By contrast TV consumption was positively associated with FM. To summarize, overweight and obese children were less fit and watched more TV than their normal weight counterparts. FM did not correspond to TEE, AEE or PAL. Muscle strength was not associated with FM in young children, but was inversely associated with FM in older children. Our cross-sectional data are consistent with the idea that increased fitness and reduced physical inactivity may prevent children from being overweight.

Blades of Lessonia vadosa (Phaeophyta) were extracted with 2% CaCl(2) solution, affording in 4.4% yield a polysaccharide which contained fucose and sulfate groups in the molar ratio 1.0:1.12. The high negative optical activity value ([alpha](D)(22)=-134.0 degrees ), FT-IR and NMR analysis suggest the presence of a fucoidan. (13)C NMR spectrum of the polysaccharide obtained by solvolytic desulfation of native fucoidan indicated the major presence of 1-->3 linked alpha-l-fucan. Depolymerization of the native fucoidan with H(2)O(2) in the presence of copper(II) acetate gave in 54.8% yield a fraction with 33.7% of sulfate content. The native fucoidan (MW 320,000) showed good anticoagulant activity whereas the radical depolymerized fraction (MW 32,000) presented a weak anticoagulant activity. These polysaccharides showed significant activation of phenylalanine-ammonia lyase (PAL), lipooxygenase (LOX) and glutathione-S-transferase (GST) defence enzyme activities in tobacco plants.

The extractable activity ofL-phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) in cell suspension cultures of bean (Phaseolus vulgaris) is greatly induced following exposure to an elicitor preparation from the cell walls of the phytopathogenic fungusColletotrichum lindemuthianum. Following exogenous application oftrans-cinnamic acid (the product of the PAL reaction) to elicitor-induced cells, the activity of the enzyme rapidly declines. Loss of enzyme activity is accompanied by inhibition of the rate of synthesis of PAL subunits, as determined by [(35)S]methionine pulse-labelling followed by specific immunoprecipitation; this is insufficient to account for the rapid loss of PAL enzyme activity. Pulse-chase and immune blotting experiments indicate that cinnamic acid does not affect the rate of degradation of enzyme subunits, but rather mediates inactivation of the enzyme. A non-dialysable factor from cinnamicacid-treated bean cells stimulates removal of PAL activity from enzyme extracts in vitro; this effect is dependent on the presence of cinnamic acid. Such loss of enzyme activity in vitro is accompanied by an apparent loss or reduction of the dehydroalanine residue of the enzyme's active site, as detected by active-site-specific tritiation, although levels of immunoprecipitable enzyme subunits do not decrease. Furthermore, cinnamic-acid-mediated loss of enzyme activity in vivo is accompanied, in pulse-chase experiments, by a greater relative loss of(35)S-labelled enzyme subunits precipitated by an immobilised active-site affinity ligand than of subunits precipitated with anti-immunoglobulin G. It is therefore suggested that a possible mechanism for cinnamic-acid-mediated removal of PAL activity may involve modification of the dehydroalanine residue of the enzyme's active site.

Earlier studies have suggested the possible role of host autolytic enzyme in the release of progeny phage from Dp-1 infected pneumococci. Several new experiments described here reinforce this notion. Specifically, the resistance of an autolysis-defective mutant to infection at low phage to cell ratios could be eliminated by prior 'coating' of the host bacteria with pneumococcal autolysin isolated from wild-type cells. Similar, productive infection was also possible by lowering the temperature of incubation to 30 degrees C, a condition that leads to a partial activation of the thermosensitive residual autolysin in the mutant cells. Other experiments, however, clearly indicate the role of the newly discovered phage-associated lysin (PAL), reported in the accompanying communication, in bacteriophage release and culture lysis; specifically, lysis was stimulated by reducing agents and inhibited by cardiolipin. It seems that both the host-related and the PAL activities are involved with Dp-1 induced lysis of pneumococci.

OBJECTIVE

To compare the Danish Physical Activity Questionnaire (DPAQ) estimating physical activity energy expenditure (PAEE) and physical activity level (PAL) and the pattern of physical activity (including health-related physical activity) with measurements from a validated position and motion instrument (ActiReg).

METHODS

One hundred and thirty-eight healthy Danish volunteers aged 20-59 years participated. Participants filled in the DPAQ at the end of the day every day for seven consecutive days and carried the ActiReg on the same days as the DPAQ were filled in.

RESULTS

A small, but statistically significant difference was seen between the DPAQ and the ActiReg for PAEE (mean: -0.32 MJ x d(-1); 95% limits of agreement: ((-2.88)-2.24 MJ x d(-1)); P = 0.003) and PAL (-0.03; ((-0.37)-0.31); P = 0.02) for the whole group and for women (P < 0.008 for PAEE and PAL), but not for men. The correlation between methods was good for PAEE (r = 0.71, P < 0.001) and PAL (r = 0.64, P < 0.001). No difference was observed for time spent in moderate plus vigorous physical activity (MVPA) (P = 0.40). Time reported in MVPA with the DPAQ was positively correlated with time spent in MVPA measured by the ActiReg (p = 0.53, P < 0.001).

CONCLUSIONS

Although the volunteer sample may influence the representativeness of the results, the DPAQ provided a close estimate of PAEE, PAL (2-5% underestimation) and health-related physical activity (MVPA) in healthy adults at group level. The results indicate that the questionnaire can be used to rank individuals according to energy expenditure and level of total physical activity and to provide information on health-related physical activity.

BACKGROUND

Many, but not all, studies have found a correlation between environmental tobacco smoke (ETS) and acute otitis media (AOM) and other adverse otologic outcomes. Given its high personal and societal costs and the divergent findings of the effect of ETS on middle ear disease, the aim of the current study was to assess the impact and possible determinant factors of ETS on recurrent (two or more) episodes of AOM.

METHODS

The study was performed at Heim Pal Children's Hospital, Ear, Nose and Throat (ENT) Department, Budapest, Hungary. Caregivers of a convenience sample of 412 children attending the ENT outpatient clinic were surveyed via a 22-item questionnaire regarding demographics, socioeconomics, and smoking behaviours of the child's family; as well as care-givers' self report of the number of AOM episodes of the child.

RESULTS

Of the 412 participants, 155 (38%) children's parents smoked. In bivariate analysis, two or more episodes of AOM correlated with reported hearing problems, day care enrolment, parental employment and increased age of the child. In multivariate logistic regression, parental smoking more than doubled a child's risk for recurrent AOM while increased maternal employment (e.g. part-time or full-time versus unemployed) boosted risk up to fourfold. Among children whose parents smoked, half-packs of cigarettes smoked per day and day care attendance doubled or nearly tripled, respectively, the risk of recurrent AOM episodes.

CONCLUSIONS

Childhood exposure to ETS is high among an ENT clinic population of Hungarian children. Such exposure correlates with AOM episodes, ENT operations and conductive hearing loss. Data such as these argue for strict laws smoke-free laws not only in Hungary, but also in Europe and around the world.

Forage quality of maize is influenced by both the content and structure of lignin in the cell wall. Phenylalanine Ammonia-Lyase (PAL) catalyzes the first step in lignin biosynthesis in plants; the deamination of L-phenylalanine to cinnamic acid. Successive enzymatic steps lead to the formation of three monolignols, constituting the complex structure of lignin. We have cloned and sequenced a PAL genomic sequence from 32 maize inbred lines currently employed in forage maize breeding programs in Europe. Low nucleotide diversity and excessive linkage disequilibrium (LD) was identified at this PAL locus, possibly reflecting selective constrains resulting from PAL being the first enzyme in the monolignol, and other, pathways. While the association analysis was affected by extended LD and population structure, several individual polymorphisms were associated with neutral detergent fiber (not considering population structure) and a single polymorphism was associated with in vitro digestibility of organic matter (considering population structure).