BACKGROUND

abdominal aortic aneurysm (AAA) distensibility may be an independent predictor of growth and rupture, possibly because it reflects changes in aortic wall structure and composition.

OBJECTIVE

to determine whether AAA distensibility is related to circulating markers of elastin and collagen metabolism.

METHODS

sixty-two male patients of median age (IQR) 68 (65-72) years with asymptomatic AAA of median (IQR) diameter 42 (37-45) mm were prospectively studied. Pressure-strain elastic modulus (Ep) and stiffness (beta) were measured using an ultrasonic echo-tracker (Diamove). Serum elastin peptides (SEP), plasma elastin-alpha1-antitrypsin complex (E-AT), procollagen III-N-terminal propeptide (PIIINP) were measured by enzyme-linked immunoassay.

RESULTS

age and smoking adjusted Ep and beta were significantly inversely related to SEP (r=-0.33 and r=-0.31 respectively, both p<0.02) and E-AT (r=-0.27 and r=-0.27 respectively, both p<0.05) both of which indicate elastolysis. By contrast, there was a significant positive correlation between PIIINP, indicative of increased collagen turn-over, and both Ep and beta (both r=0.45, p<0.01 unadjusted correlations).

CONCLUSIONS

increased elastolysis is associated with increased AAA wall distensibility; whereas increased collagen turn-over is associated with reduced distensibility.

We measured CSF and plasma contents of beta-endorphin (beta-EP), beta-lipotropin (beta-LPH), and ACTH in 24 patients with Parkinson's disease; 14 had not been treated. CSF beta-EP concentrations in untreated patients were lower than in 15 controls (p less than 0.005), but values did not differ significantly in treated and untreated patients. In untreated and treated patients, ACTH and beta-LPH CSF, and beta-EP, beta-LPH, and ACTH plasma concentrations were in the same range as controls. The Parkinson's disease-related decrease of CSF beta-EP levels further supports the concept that there is a generalized brain disorder in Parkinson's disease affecting more than dopaminergic neurons in the substantia nigra.

Decoding the molecular composition of individual Ngn3 + endocrine progenitors (EPs) during pancreatic morphogenesis could provide insight into the mechanisms regulating hormonal cell fate. Here, we identify population markers and extensive cellular diversity including four EP subtypes reflecting EP maturation using high-resolution single-cell RNA-sequencing of the e14.5 and e16.5 mouse pancreas. While e14.5 and e16.5 EPs are constantly born and share select genes, these EPs are overall transcriptionally distinct concomitant with changes in the underlying epithelium. As a consequence, e16.5 EPs are not the same as e14.5 EPs: e16.5 EPs have a higher propensity to form beta cells. Analysis of e14.5 and e16.5 EP chromatin states reveals temporal shifts, with enrichment of beta cell motifs in accessible regions at later stages. Finally, we provide transcriptional maps outlining the route progenitors take as they make cell fate decisions, which can be applied to advance the in vitro generation of beta cells.

OBJECTIVE

To compare efficacy between double-dose methotrexate and single-dose methotrexate for treatment of tubal ectopic pregnancy (EP).

METHODS

Between March 2008 and February 2011,157 patients who had tubal EP diagnosed by a non-laparoscopic approach and were hemodynamically stable were enrolled in a prospective study in Qassim, Saudi Arabia. The participants were randomized to receive either double-dose (50mg/m(2) intramuscularly on days 0 and 4; group 1) or single-dose (50mg/m(2) intramuscularly on day 0; group 2) methotrexate. Serum human chorionic gonadotropin (β-hCG) levels were followed until negative.

RESULTS

The overall success rate was comparable between groups 1 and 2 (88.6% versus 82.0%, P=0.1). The duration of follow up until negative β-hCG was shorter in group 1 (P=0.001). Receiver operative characteristics showed that higher cut-off levels of β-hCG and gestational mass diameter were associated with successful outcome in group 1. Among participants with initial β-hCG of 3600-5500 mIU/mL, the success rate was higher in group 1 (P=0.03). There was no significant difference between groups in adverse effects.

CONCLUSIONS

For treatment of EP, double-dose methotrexate had efficacy and safety comparable to that of single-dose methotrexate; it had better success among patients with moderately high β-hCG and led to a shorter follow up.

OBJECTIVE

The efficacy of neurofeedback as a treatment for attention-deficit/hyperactivity disorder (ADHD), and whether neurofeedback is a viable alternative for stimulant medication, is still an intensely debated subject. The current randomized controlled trial compared neurofeedback to (1) optimally titrated methylphenidate and (2) a semi-active control intervention, physical activity, to account for nonspecific effects.

METHODS

A multicenter 3-way parallel-group study with balanced randomization was conducted. Children with a DSM-IV-TR diagnosis of ADHD, aged 7-13 years, were randomly allocated to receive neurofeedback (n = 39), methylphenidate (n = 36), or physical activity (n = 37) over a period of 10-12 weeks. Neurofeedback comprised theta/beta training on the vertex (Cz). Physical activity consisted of moderate to vigorous intensity exercises. Neurofeedback and physical activity were balanced in terms of number (~30) and duration of sessions. A double-blind pseudorandomized placebo-controlled crossover titration procedure was used to determine an optimal dose in the methylphenidate intervention. Parent and teacher ratings on the Strengths and Difficulties Questionnaire (SDQ) and Strengths and Weaknesses of ADHD Symptoms and Normal Behavior (SWAN) were used to assess intervention outcomes. Data collection took place between September 2010 and March 2014.

RESULTS

Intention-to-treat analyses revealed an improvement in parent-reported behavior on the SDQ and the SWAN Hyperactivity/Impulsivity scale, irrespective of received intervention (ηp² = 0.21-0.22, P ≤ .001), whereas the SWAN Inattention scale revealed more improvement in children who received methylphenidate than neurofeedback and physical activity (ηp² = 0.13, P ≤ .001). Teachers reported a decrease of ADHD symptoms on all measures for methylphenidate, but not for neurofeedback or physical activity (range of ηp² = 0.14-0.29, P < .001).

CONCLUSIONS

The current study found that optimally titrated methylphenidate is superior to neurofeedback and physical activity in decreasing ADHD symptoms in children with ADHD.

BACKGROUND

ClinicalTrials.gov identifier: NCT01363544.

BACKGROUND

Aortic compliance, as measured by the pressure-strain elastic modulus (Ep) and stiffness (B), may allow a more precise estimate of abdominal aortic aneurysm rupture risk than size alone.

OBJECTIVE

To determine the relationships between AAA compliance, size, growth, and clinical outcome.

METHODS

One-hundred and twelve patients with initially non-operated AAA (86 men, 26 women, mean age 73 years), recruited from five centres, underwent baseline compliance measurements and were then followed for a median of 7 (range 2-18) months; 85 patients underwent repeated measurements (median 3, range 2-5) 3-6-monthly over a median of 12 (range 3-18 months).

RESULTS

Seven patients have ruptured and 16 have undergone repair of non-ruptured AAA. AAA that ruptured had significantly lower Ep and B (more compliant). In AAA that ruptured or required repair there was an inverse relationship between diameter and Ep and B. In those undergoing repeated measurements AAA expansion was only associated with a significant increase in Ep and B in non-operated patients.

CONCLUSIONS

Baseline AAA compliance was significantly related to rupture and the future requirement for operative repair. Failure of compliance to increase with size may be a marker for rapid growth, developmental symptoms and rupture.

OBJECTIVE

This multicentre prospective randomised trial was undertaken to evaluate the usefulness of an electrophysiological study (EPS)-guided/implantable cardioverter defibrillator (ICD) strategy in patients at high risk of sudden death (SD) early after myocardial infarction (MI). Previous studies have shown the benefits of such a strategy only in high-risk patients late after MI.

RESULTS

We enrolled 143 survivors of acute MI (<1 month) with left ventricular ejection fraction < or = 35% and either frequent >> or =10/h) premature ventricular complexes (PVCs), or depressed heart rate variability (SDNN < 70 ms) or abnormal signal-averaged ECG, who were able to tolerate optimised beta-blocker therapy (68 +/- 40 mg/day of metoprolol). Of these, 138 were randomised, in a 2:3 ratio, to two therapeutic strategies: conventional (CONV) strategy (n = 59) or EPS-guided/ICD strategy (n = 79). The latter resulted in ICD implantation in 24 inducible patients and in CONV therapy in the remaining 55. During a mean follow-up of 540 +/- 378 days, 26 patients (19%) died: nine (6.5%) SD, nine (6.5%) non-SD, and four (3%) non-cardiac death; in four patients (3%) the cause of death was unknown. The actuarial overall mortality for the CONV and EPS-guided/ICD arms was 18% vs 14% after 1 year and 29.5% vs 20% after 2 years, respectively (P = 0.3 and 0.2).

CONCLUSIONS

Despite optimal therapy, mortality remains significant in high-risk patients following MI. Although there is a trend in favour of EPS-guided/ICD, our data are insufficient to demonstrate a survival benefit of this strategy early after MI.

The population of brain pericytes, a cell type important for vessel stability and blood brain barrier function, has recently been shown altered in patients with Alzheimer's disease (AD). The underlying reason for this alteration is not fully understood, but progressive accumulation of the AD characteristic peptide amyloid-beta (Aβ) has been suggested as a potential culprit. In the current study, we show reduced number of hippocampal NG2+ pericytes and an association between NG2+ pericyte numbers and Aβ1-40 levels in AD patients. We further demonstrate, using in vitro studies, an aggregation-dependent impact of Aβ1-40 on human NG2+ pericytes. Fibril-EP Aβ1-40 exposure reduced pericyte viability and proliferation and increased caspase 3/7 activity. Monomer Aβ1-40 had quite the opposite effect: increased pericyte viability and proliferation and reduced caspase 3/7 activity. Oligomer-EP Aβ1-40 had no impact on either of the cellular events. Our findings add to the growing number of studies suggesting a significant impact on pericytes in the brains of AD patients and suggest different aggregation forms of Aβ1-40 as potential key regulators of the brain pericyte population size.

Theileria equi (Laveran 1901) and Babesia caballi (Nuttall and Strickland 1910) are the causative agents of Equine Piroplasmosis (EP), a severe and problematic disease compromising international movement of horses. Infected horses usually become asymptomatic carriers and, for this reason, their movement across borders may become restricted. The aim of this study was to assess the seroprevalence of EP in Southern France and to evaluate risk factors associated with these parasites. In 2002, we performed a complement fixation test (CF) with blood samples from 443 horses stabled at 95 different farms located in the region of Camargue. Two epidemiological questionnaires have been used: one for each single horse (individual and management factors) and one for each place where horses were sampled (environment, presence of other species, etc.) to identify risk factors for seropositivity. T. equi and B. caballi had a seroprevalence of 58 % and 12.9%, respectively. For T. equi, sex, age, activity, management, and living with or near cattle were identified as risk factors, while for B. caballi, only living in wetlands was recognized as a risk factor in the bivariate analysis. In the multivariate analysis, the best model for T. equi included as variables age, breed, and deworming, while the best model for B. caballi included the type of housing during day and the contact with cows.

Biofilm is a complex aggregate of cells that adhere to each other and produce an extracellular matrix. In Bacillus subtilis, an extracellular polysaccharide (EPS) and amyloid fiber (TasA), synthesized by the epsA-epsO and tapA-sipW-tasA operons, respectively, are the primary components of the extracellular matrix. In the current study, we investigated the functional role of the previously uncharacterized veg gene in B. subtilis. Overproduction of Veg, a small protein highly conserved among Gram-positive bacteria, stimulated biofilm formation via inducing transcription of the tapA-sipW-tasA operon. Moreover, overproduced Veg restored the impairment of biofilm formation in mutants carrying a deletion of of sinI, slrA, or slrR, encoding an antirepressor of SinR that acts as the master regulator of biofilm formation, while biofilm morphology in the absence of SinR was not affected by either additional veg deletion or overproduction, indicating that Veg negatively regulates SinR activity independently of the known antirepressors. Expression of sinR was not affected in Veg-overproducing cells, and amounts of SinR were similar in cells expressing different levels of Veg, strongly suggesting that Veg modulates the repressor activity of SinR. Interestingly, the results of in vivo pulldown assays of the SinR complex indicate that Veg inhibits the interactions between SinR and SlrR. Based on these findings, we propose that Veg or a Veg-induced protein acts as an antirepressor of SinR to regulate biofilm formation.

The effect of estradiol and progesterone therapy in serum and liver on the lipid profile of naturally menopausal albino rats of the Wistar strain of different age groups (12,18 and 24 months) have been measured and compared with the age matched groups. Three months old rats were used as young controls. The aged rats were administered subcutaneous injection of 17-beta-estradiol (0.1 microg/g body weight), progesterone (2.5 microg/g body weight) and similar concentrations of both in combined treatment for 1 month and the level of triglycerides (TG), total lipids (TL), total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL) and very low density lipoprotein (VLDL) were measured in serum and liver of 3, 12, 18 and 24 months old control as well as treated groups. The results show that TG, HDL, VLDL levels were increased significantly by 71%, 155%, 54%, respectively in liver of 24 months old rats by combination treatment when compared with age matched control animals. The levels of TL, TC and LDL were decreased by 20%, 31%, and 30%, respectively in serum of 12 months old rats in combination treatment group. The effect was more significant in 12 and 24 months old female rats with administration of estrogen and combined (EP) treatments. The results indirectly suggest that hormone replacement therapy (HRT) can reduce the risk of cardiovascular disease (CVD) thereby playing a cardio-protective role by restoring lipid and hormone levels to the similar levels as found in young female animals.

The influence exerted by somatostatin on the secretion of ACTH and opioid peptides has still to be clarified. To gain further information on this issue, we performed in 10 normal volunteers two CRF tests (100 micrograms i.v.) one of which was preceded by s.c. injection of 100 micrograms of the long-acting somatostatin analogue SMS 201-995 (Sandostatin, Sandoz) (SMS), given 30 minutes before CRF. Premedication with SMS markedly inhibited the response of beta-EP to CRF, leaving unchanged the response of beta-LPH, ACTH and cortisol; mean incremental areas of beta-EP were 199.8 +/- 49.31 (SEM) vs 532.9 +/- 95.91 pmol 120 min (P less than 0.01) in the CRF test with and without SMS, respectively. To interpret the selective inhibitory effect of SMS on CRF-stimulated beta-EP secretion, it can be hypothesized that: a) the action of SMS was confined to a population of pituicytes preferentially secreting beta-EP; b) SMS interfered with the processing of POMC inhibiting the formation of beta-EP; c) SMS acted on extrapituitary, possibly peripheral, sources of beta-EP. In conclusion, this study indicates that, in man, somatostatin selectively inhibits the CRF-induced secretion of beta-EP, but not that of ACTH and beta-LPH, by an action that may be exerted at pituitary or extrapituitary level. This is a further example of dissociated secretion of POMC-derived peptides.

Salt stress is one of the most serious factors limiting the productivity of rice, the staple diet in many countries. Gibberellic acid has been reported to reduce NaCl-induced growth inhibition in some plants including rice. Most paddy soils have a natural population of Cyanobacteria, prokaryotic photosynthethic microorganisms, which synthesize and liberate plant growth regulators such as gibberellins that could exert a natural beneficial effect on salt stressed rice plants. The aim of this work was to evaluate the effect of the cyanobacterium Scytonema hofmanni extracellular products on the growth of rice seedlings inhibited by NaCl and to compare it with the effect of the gibberellic acid in the same stress condition. Growth (length and weight of the seedlings) and biochemical parameters (5-aminolevulinate dehydratase activity, total free porphyrin and pigments content) were evaluated. Salt exposure negatively affected all parameters measured, with the exception of chlorophyll. Chlrorophyll concentrations nearly doubled upon exposure to high salt. Gibberellic acid counteracted the effect of salt on the length and dry weight of the shoot, and on carotenoid and chlorophyll b contents. Extracellular products nullified the salt effect on shoot dry weight and carotenoid content; partially counteracted the effect on shoot length (from 54% to 38% decrease), root dry weight (from 59% to 41% decrease) and total free porphyrin (from 31 to 13% decrease); reduced by 35% the salt increase of chlorophyll a; had no effect on root length and chlorophyll b. Gibberellic acid and extracellular products increased 5-aminolevulinate dehydratase activity over the control without salt. When coincident with high salinity, exposure to either EP or GA3, resulted in a reversal of shoot-related responses to salt stress. We propose that Scytonema hofmanni extracellular products may counteract altered hormone homeostasis of rice seedlings under salt stress by producing gibberellin-like plant growth regulators.

Gram-positive bacteria can cause various infections including hospital-acquired infections. While in the biofilm, the resistance of bacteria to both antibiotics and the human immune system is increased causing difficulties in the treatment. Bacillus subtilis, a non-pathogenic Gram-positive bacterium, is widely used as a model organism for studying biofilm formation. Here we investigated the effect of novel synthesized chloro- and bromo-containing 2(5H)-furanones on biofilm formation by B. subtilis. Mucobromic acid (3,4-dibromo-5-hydroxy-2(5H)-furanone) and the two derivatives of mucochloric acid (3,4-dichloro-5-hydroxy-2(5H)-furanone)-F8 and F12-were found to inhibit the growth and to efficiently prevent biofilm formation by B. subtilis. Along with the low production of polysaccharide matrix and repression of the eps operon, strong repression of biofilm-related yqxM also occurred in the presence of furanones. Therefore, our data confirm that furanones affect significantly the regulatory pathway(s) leading to biofilm formation. We propose that the global regulator, Spo0A, is one of the potential putative cellular targets for these compounds.

One of the most important postharvest plant pathogens that affect strawberries, grapes and tomatoes is Botrytis cinerea, known as gray mold. The fungus remains in latent form until spore germination conditions are good, making infection control difficult, causing great losses in the whole production chain. This study aimed to purify and identify phenazine-1-carboxylic acid (PCA) produced by the Pseudomonas aeruginosa LV strain and to determine its antifungal activity against B. cinerea. The compounds produced were extracted with dichloromethane and passed through a chromatographic process. The purity level of PCA was determined by reversed-phase high-performance liquid chromatography semi-preparative. The structure of PCA was confirmed by nuclear magnetic resonance and electrospray ionization mass spectrometry. Antifungal activity was determined by the dry paper disk and minimum inhibitory concentration (MIC) methods and identified by scanning electron microscopy and confocal microscopy. The results showed that PCA inhibited mycelial growth, where MIC was 25 μg mL-1. Microscopic analysis revealed a reduction in exopolysaccharide (EPS) formation, showing distorted and damaged hyphae of B. cinerea. The results suggested that PCA has a high potential in the control of B. cinerea and inhibition of EPS (important virulence factor). This natural compound is a potential alternative to postharvest control of gray mold disease.

The present study aims to discover single nucleotide polymorphisms (SNPs) at the apelin gene (APLN) in relation to arterial stiffness, and to explore its molecular mechanisms. A two-step genetic association study was conducted using 799 and 937 subjects in the screening and validation data, respectively. Four tagging SNPs of APLN were tested. SNP rs3115757 was significantly associated with stiffness parameters (β, Ep and PWV) in women, but not in men. The function of rs3115757 was tagged by rs3115758 which is located in miR-765 binding site in the 3' untranslated region of APLN. The reporter assay confirmed that different alleles of rs3115758 interfered with miR-765 binding and then modified APLN expression. Over-expression of miR-765 in endothelial cells decreased mRNA and protein levels of APLN, which further inhibited the phosphorylation of eNOS and ERK/Akt/AMPK signaling. Collectively, our data showed that rs3115758 accounts for the susceptibility of arterial stiffening through miR-765-induced APLN repression.

Understanding bacterial adhesion to surfaces requires knowledge of the forces that govern bacterial-surface interactions. Biofilm formation on stainless steel 316 (SS316) by three bacterial species was investigated by examining surface force interaction between the cells and metal surface using atomic force microscopy (AFM). Bacterial-metal adhesion force was quantified at different surface delay time from 0 to 60s using AFM tip coated with three different bacterial species: Gram-negative Massilia timonae and Pseudomonas aeruginosa, and Gram-positive Bacillus subtilis. The results revealed that bacterial adhesion forces on SS316 surface by Gram-negative bacteria is higher (8.53±1.40 nN and 7.88±0.94 nN) when compared to Gram-positive bacteria (1.44±0.21 nN). Physicochemical analysis on bacterial surface properties also revealed that M. timonae and P. aeruginosa showed higher hydrophobicity and surface charges than B. subtilis along with the capability of producing extracellular polymeric substances (EPS). The higher hydrophobicity, surface charges, and greater propensity to form EPS by M. timonae and P. aeruginosa led to high adhesive force on the metal surface.

Extracellular polymeric substances (EPS) secreted by suspended cultures of microorganisms from an activated sludge plant in the presence of glucose were characterized in detail using colorimetry, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. EPS produced by the multi-species community were similar to literature reports of pure cultures in terms of functionalities with respect to C and O but differed subtly in terms of N and P. Hence, it appears that EPS produced by different microorganisms maybe homologous in major chemical constituents but may differ in minor components such as lipids and phosphodiesters. The role of specific EPS constituents on microbial aggregation was also determined. The weak tendency of microorganisms to bioflocculate during the exponential growth phase was attributed to electrostatic repulsion when EPS concentration was low and acidic in nature (higher fraction of uronic acids to total EPS) as well as reduced polymer bridging. However, during the stationary phase, polymeric interactions overwhelmed electrostatic interactions (lower fraction of uronic acids to total EPS) resulting in improved bioflocculation. More specifically, microorganisms appeared to aggregate in the presence of protein secondary structures including aggregated strands, beta-sheets, alpha- and 3-turn helical structures. Bioflocculation was also favored by increasing O-acetylated carbohydrates and overall C-(O,N) and O=C-OH+O=C-OR functionalities.

<A<em>b</em>stractText>To investigate the effects of a 16-week concurrent exercise regimen [resistance exercise (RE) + functional exercise (FE)] in com<em>b</em>ination with, or without, a leucine-enriched whey protein isolate supplement on muscle strength, physical functioning, aero<em>b</em>ic capacity, and cardiometa<em>b</em>olic health in older adults (≥60 years). Physical activity levels were also evaluated 6 months post-cessation of the intervention.</A<em>b</em>stractText><p><div>(<em>b</em>)Methods</<em>b</em>)</div>Forty-six, community-dwelling, previously untrained males, and females [age: 68 ± 5 years (mean ± SD); BMI: 27.8 ± 6.2 kg/m²] who completed the trial were initially randomized to one of two independent arms [Exercise <i>n</i> = 24 (E); Exercise+Protein <i>n</i> = 22 (<em>EP</em>)]. Both arms completed 16 weeks of RE (performed to fatigue) (2 times/week) with FE (1 time/week) on non-consecutive days. Additionally, <em>EP</em> were administered a leucine-enriched whey protein supplement (3 times/day) for 16 weeks <em>b</em>ased on individual <em>b</em>ody-weight (1.5 g/kg/day).</p><p><div>(<em>b</em>)Results</<em>b</em>)</div>As a result of dietary supplementation, protein intake increased in <em>EP</em> (∼1.2 ± 0.4 to 1.5 ± 0.7 g/kg/day) during the intervention. Maximal strength (1RM) values for leg press (E: +39 ± 7 kg, <i>p</i> = 0.006; <em>EP</em>: +63 ± 7 kg, <i>p</i> < 0.001), chest press (E: +22 ± 4 kg, <i>p <</i> 0.001; <em>EP</em>: +21 ± 6 kg, <i>p</i> < 0.001), and <em>b</em>icep curl (E: +7 ± 0 kg, <i>p</i> = 0.002; <em>EP</em>: +6 ± 1 kg, <i>p</i> = 0.008) significantly increased in E and <em>EP</em> respectively, with no differences <em>b</em>etween arms (<i>p</i> > 0.05). Physical functioning in the o<em>b</em>stacle course (E: -5.1 ± 6.8 s, <i>p</i> < 0.001; <em>EP</em>: -2.8 ± 0.8 s, <i>p</i> < 0.001) and short-physical performance <em>b</em>attery scores (E: +0.5 ± 0.5, <i>p</i> = <0.001; <em>EP</em>: +0.4 ± 0.5, <i>p</i> = 0.038), and aero<em>b</em>ic capacity in the 6-min walk test (E: +37 ± 24 m, <i>p =</i> 0.014; <em>EP</em>: +36 ± 3 m, <i>p</i> = 0.005) improved in E and <em>EP</em> respectively, with no differences <em>b</em>etween arms (<i>p</i> > 0.05). No significant change was o<em>b</em>served for markers of cardiometa<em>b</em>olic health (glycaemic control or <em>b</em>lood pressure) (<i>p</i> > 0.05). At follow-up, 86% of older adults reported to performing physical activity ≥1 per week. Of those, 61% were still participating in strength- and cardiovascular- <em>b</em>ased exercise.</p><A<em>b</em>stractText>Concurrent exercise (RE + FE) offers a potent method to com<em>b</em>at age-related muscle weakness, and our results suggest a high proportion of older adults may continue to exercise unsupervised. However, leucine-enriched whey protein isolate supplementation did not confer any additional <em>b</em>enefit in those already consuming ample amounts of dietary protein at trial enrolment. Future trials should utilize a whole-foods approach and investigate the effects in frail and non-frail older adults ha<em>b</em>itually consuming the RDA of protein, to assess if a higher intake of protein is needed to delay the onset of muscle weakness.</A<em>b</em>stractText><A<em>b</em>stractText>Clinicaltrials.gov Identifier: NCT02912130.</A<em>b</em>stractText>

BACKGROUND

Bifidobacterium longum 105-A produces markedly high amounts of capsular polysaccharides (CPS) and exopolysaccharides (EPS) that should play distinct roles in bacterial-host interactions. To identify the biological function of B. longum 105-A CPS/EPS, we carried out an informatics survey of the genome and identified the EPS-encoding genetic locus of B. longum 105-A that is responsible for the production of CPS/EPS. The role of CPS/EPS in the adaptation to gut tract environment and bacteria-gut cell interactions was investigated using the ΔcpsD mutant.

RESULTS

A putative B. longum 105-A CPS/EPS gene cluster was shown to consist of 24 putative genes encoding a priming glycosyltransferase (cpsD), 7 glycosyltransferases, 4 CPS/EPS synthesis machinery proteins, and 3 dTDP-L-rhamnose synthesis enzymes. These enzymes should form a complex system that is involved in the biogenesis of CPS and/or EPS. To confirm this, we constructed a knockout mutant (ΔcpsD) by a double cross-over homologous recombination. Compared to wild-type, the ∆cpsD mutant showed a similar growth rate. However, it showed quicker sedimentation and formation of cell clusters in liquid culture. EPS was secreted by the ∆cpsD mutant, but had altered monosaccharide composition and molecular weight. Comparison of the morphology of B. longum 105-A wild-type and ∆cpsD by negative staining in light and electron microscopy revealed that the formation of fimbriae is drastically enhanced in the ∆cpsD mutant while the B. longum 105-A wild-type was coated by a thick capsule. The fimbriae expression in the ∆cpsD was closely associated with the disappearance of the CPS layer. The wild-type showed low pH tolerance, adaptation, and bile salt tolerance, but the ∆cpsD mutant had lost this survivability in gastric and duodenal environments. The ∆cpsD mutant was extensively able to bind to the human colon carcinoma Caco-2 cell line and was phagocytosed by murine macrophage RAW 264.7, whereas the wild-type did not bind to epithelial cells and totally resisted internalization by macrophages.

CONCLUSIONS

Our results suggest that CPS/EPS production and fimbriae formation are negatively correlated and play key roles in the survival, attachment, and colonization of B. longum 105-A in the gut.

Rapsyn, a complex postsynaptic protein of the striated muscle, assembles acetylcholine receptors (AChR) at high density at the motor endplate (EP). Neuromuscular junctions of mice lacking rapsyn show no clusters of AChRs or other structural postsynaptic proteins such as beta-dystroglycan and utrophin. Humans with mutations in the rapsyn gene ( RAPSN) are affected with a postsynaptic form of congenital myasthenic syndrome (CMS) characterized by impairment of the morphologic development of the postsynaptic region. We have identified four patients from four different families with RAPSNmutations and CMS, confirmed in two cases by microelectrode and electron microscopy studies. The N88K mutation was present in all patients. One patient who was homozygous for N88K was only mildly affected, while the other three patients who were heterozygous for N88K and a second mutation (either L14P, 46insC, or Y269X) were severely affected. Mutations 46insC and Y269X predicts truncation of the protein. L14P predicts a conformational change at the N-terminus that may disrupt membrane association. N88K occurs within the putative leucine zipper motif potentially important for AChR clustering. These findings may explain the severe clinical involvement of compound heterozygous patients.

<A<em>b</em>stractText>Administration of endogenous mediators or exogenous chemicals in migraine patients provoke early headaches and delayed migraine-like attacks. Although migraine provoking su<em>b</em>stances are normally vasodilators, dilation of arterial vessels does not seem to <em>b</em>e the sole contri<em>b</em>uting factor, and the underlying mechanisms of the delayed migraine pain are mostly unknown. Sustained mechanical allodynia is a common response associated with the local administration of various proalgesic su<em>b</em>stances in experimental animals and humans. Here, we investigated the a<em>b</em>ility of a series of endogenous mediators which provoke or do not provoke migraine in patients, to cause or not cause mechanical allodynia upon their injection in the mouse perior<em>b</em>ital area.</A<em>b</em>stractText><A<em>b</em>stractText>Mechanical allodynia was assessed with the von Frey filament assay. Stimuli were given <em>b</em>y su<em>b</em>cutaneous injection in the perior<em>b</em>ital area of C57BL/6J mice; antagonists were administered <em>b</em>y local and systemic injections.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>Calcitonin gene related peptide (CGRP), <em>b</em>ut not adrenomedullin and amylin, pituitary adenylyl cyclase activating peptide (PACAP), <em>b</em>ut not vasoactive intestinal polypeptide (VIP), histamine, prostaglandin E<su<em>b</em>)2</su<em>b</em>) (PGE<su<em>b</em>)2</su<em>b</em>)) and prostacyclin (PGI<su<em>b</em>)2</su<em>b</em>)), <em>b</em>ut not PGF<su<em>b</em>)2α,</su<em>b</em>) evoked a dose-dependent perior<em>b</em>ital mechanical allodynia. The painful responses were attenuated <em>b</em>y systemic or local (perior<em>b</em>ital) administration of antagonists for CGRP (CLR/RAMP1), PACAP (PAC-1), histamine H<su<em>b</em>)1</su<em>b</em>), PGE<su<em>b</em>)2</su<em>b</em>) (<em>EP</em><su<em>b</em>)4</su<em>b</em>)), and PGI<su<em>b</em>)2</su<em>b</em>) (IP) receptors, respectively.</p><p><div>(<em>b</em>)CONCLUSIONS</<em>b</em>)</div>The correspondence <em>b</em>etween su<em>b</em>stances that provoke (CGRP; PACAP, histamine, PGE<su<em>b</em>)2</su<em>b</em>), PGI<su<em>b</em>)2</su<em>b</em>)), or do not provoke (VIP and PGF<su<em>b</em>)2α</su<em>b</em>)), migraine-like attacks in patients and perior<em>b</em>ital allodynia in mice suggests that the study of allodynia in mice may provide information on the proalgesic mechanisms of migraine-provoking agents in humans. Results underline the a<em>b</em>ility of migraine-provoking su<em>b</em>stances to initiate mechanical allodynia <em>b</em>y acting on peripheral terminals of trigeminal afferents.</p>

Interleukin-4 (IL-4), which was originally identified as a B-cell growth factor, has been shown to produce diverse effects on hemopoietic progenitors. The present study investigated the effects of purified recombinant murine IL-4 on early hemopoetic progenitors in methylcellulose culture. IL-4 supported the formation of blast cell colonies and small granulocyte/macrophage (GM) colonies in cultures of marrow and spleen cells of normal mice as well as spleen cells of mice treated with 150 mg/kg 5-fluorouracil (5-FU) 4 days earlier. When the blast cell colonies were individually picked and replated in cultures containing WEHI-3 conditioned medium and erythropoietin (Ep), a variety of colonies were seen, including mixed erythroid colonies, indicating the multipotent nature of the blast cell colonies supported by IL-4. To test whether or not IL-4 affects multipotent progenitors directly, we replated pooled blast cells in cultures under varying conditions. In the presence of Ep, both IL-3 and IL-4 supported a similar number of granulocyte/erythrocyte/macrophage/megakaryocyte (GEMM) colonies. However, the number of GM colonies supported by IL-4 was significantly smaller than that supported by IL-3. When colony-supporting abilities of IL-4 and IL-3 were compared using day-4 post-5-FU spleen and day-2 post-5-FU marrow cells, IL-4 supported the formation of fewer blast cell colonies than did IL-3. IL-4 and IL-6 revealed synergy in support of colony formation from day 2 post-5-FU marrow cells. These results indicate that murine IL-4 is another direct-acting multilineage colony-stimulating factor (multi-CSF), similar to IL-3, that acts on primitive hemopoietic progenitors.