Different profiles of T-cell IFN-gamma and IL-12 in allergen-induced early and dual responders with asthma.
IFN-gamma and IL-12 are anti-inflammatory cytokines released from various cells, including T cells. Allergen inhalation by atopic subjects with asthma results in 2 bronchoconstrictor phenotypes, termed isolated early and dual responders . Persistence of allergen-induced airway response and inflammation is a distinctive feature of dual responders. To evaluate the roles of IFN-gamma and IL-12 in resolving allergen-induced airway inflammation by comparing T lymphocytes (CD4 + and CD8 + cells) producing these cytokines in isolated early and dual responders. Twenty-four subjects with asthma (12 isolated early and 12 dual responders) were challenged with inhaled allergen. Peripheral blood and induced sputum were taken before and 1 day, 3 days, and 7 days after challenge. Frequency of IFN-gamma, IL-12, IL-4, and IL-13 producing CD4 + and CD8 + cells was assessed by using flow cytometry. After allergen, both CD4 + and CD8 + IFN-gamma positive cells in peripheral blood significantly decreased in dual responders only, whereas CD4 + and CD8 + IFN-gamma positive cells in induced sputum significantly increased in isolated early responders only. By contrast, IL-12 positive cells in peripheral blood significantly increased after allergen challenge only in isolated early responders. The ratio of CD4 + and CD8 + IL-4/IFN-gamma positive cells in peripheral blood significantly decreased in isolated early responders by 3 days and had recovered by 7 days. These results suggest that contrasting profiles of IFN-gamma and IL-12 production may be responsible for different time courses of allergen-induced airway responses between isolated early and dual responders.


Identification of bitter peptides in aged cheddar cheese.
The compounds responsible for the bitter taste of aged "sharp" Cheddar cheese were characterized. Sensory-guided fractionation techniques using gel permeation chromatography and multi-dimension semi-preparative reversed-phase high-performance liquid chromatography revealed the presence of multiple bitter compounds. The compounds with the highest perceived bitterness intensity were identified by tandem mass spectrometry de novo peptide sequencing as GPVRGPFPIIV, YQEPVLGPVRGPFPI, MPFPKYPVEP, MAPKHKEMPFPKYPVEPF, and APHGKEMPFPKYPVEPF; all originated from β-casein. Subsequent quantitative liquid chromatography-tandem mass spectrometry analysis reported that the concentrations of GPVRGPFPIIV, YQEPVLGPVRGPFPI, and MPFPKYPVEP increased during maturation by 28.7-, 3.1-, and 1.8-fold, respectively. When directly compared to young "mild" Cheddar, APHGKEMPFPKYPVEPF was reported only in the sharp Cheddar cheese, whereas the concentration of MAPKHKEMPFPKYPVEPF did not change. Further taste re-engineering sensory experiments confirmed the importance of the identified peptides to the bitterness of sharp Cheddar. The bitter intensity of the aged "sharp" Cheddar model (mild Cheddar with equivalent concentrations of the five bitter peptides in the sharp sample) was rated as not significantly different from the authentic sharp Cheddar cheese. Among the five peptides, GPVRGPFPIIV was reported to be the main contributor to the bitterness intensity of sharp Cheddar. Furthermore, a difference from control sensory test also confirmed the significance of the bitter taste to the overall perception of aged Cheddar flavor. The sharp Cheddar model was reported to be significantly more similar to aged "sharp" Cheddar in comparison to the young "mild" Cheddar cheese sample.


Alcohol Use, Discrimination, and Psychological Partner Abuse Among LGBQ+ College Students: Results From a Daily Diary Study.
Alcohol use correlates with psychological partner abuse (PA) perpetration among lesbian, gay, bisexual, queer, and other nonheterosexual (LGBQ+) young adults. However, less is known about the proximal association between alcohol use and psychological PA within this population, which would provide valuable information for intervention development. Informed by minority stress and alcohol-related PA theories, we evaluated whether (a) psychological PA perpetration odds increased as the number of drinks consumed prior to psychological PA on a given day increased, (b) psychological PA perpetration odds were greater following heavy episodic drinking (HED) relative to non-HED, and (c) experiencing LGBQ+-specific discrimination (i.e., heterosexist harassment, incivility, and hostility) strengthened the association between daily alcohol use (number of drinks, HED) and subsequent psychological PA perpetration. LGBQ+ college students (N = 41; 75.6% women, 22.0% men, and 2.4% transgender/non-binary) completed a baseline survey of past-year discrimination before completing daily reports of their alcohol use and psychological PA for 60 consecutive days. Multilevel modeling revealed that drinking more than one usually does on a given day is positively associated with subsequent psychological PA odds (OR = 1.31, p < .001). Psychological PA was more likely following HED relative to non-HED (OR = 3.23, p < .001). Unexpectedly, experiencing discrimination was negatively associated with psychological PA odds across models (OR = .26 p = .01). No alcohol × discrimination interactions emerged. Results support alcohol use as a proximal risk factor for psychological PA among LGBQ+ college students and underscore the need for more nuanced examination of discrimination and other contextual variables in alcohol-related PA.


Fusidic acid-resistant mutants of Salmonella enterica serovar Typhimurium with low fitness in vivo are defective in RpoS induction.
Mutants of Salmonella enterica serovar Typhimurium resistant to fusidic acid (Fusr) have mutations in fusA, the gene encoding translation elongation factor G (EF-G). Most Fusr mutants have reduced fitness in vitro and in vivo, in part explained by mutant EF-G slowing the rate of protein synthesis and growth. However, some Fusr mutants with normal rates of protein synthesis still suffer from reduced fitness in vivo. As shown here, Fusr mutants could be similarly ranked in their relative fitness in mouse infection models, in a macrophage infection model, in their relative hypersensitivity to hydrogen peroxide in vivo and in vitro, and in the amount of RpoS production induced upon entry into the stationary phase. We identify a reduced ability to induce production of RpoS (sigmas) as a defect associated with Fusr strains. Because RpoS is a regulator of the general stress response, and an important virulence factor in Salmonella, an inability to produce RpoS in appropriate amounts can explain the low fitness of Fusr strains in vivo. The unfit Fusr mutants also produce reduced levels of the regulatory molecule ppGpp in response to starvation. Because ppGpp is a positive regulator of RpoS production, we suggest that a possible cause of the reduced levels of RpoS is the reduction in ppGpp production associated with mutant EF-G. The low fitness of Fusr mutants in vivo suggests that drugs that can alter the levels of global regulators of gene expression deserve attention as potential antimicrobial agents.


Box-Behnken design for removal of uranium(VI) from aqueous solution using poly(ethylene glycol) based dicationic ionic liquid impregnated chitosan.
Poly(ethylene glycol) bis(methylimidazolium) di[bis(trifluoromethylsulfonyl)imide] was synthesized as an ionic liquid and impregnated onto chitosan. The removal of uranium(VI) ions from aqueous solution was investigated with batch sorption tests using ionic liquid impregnated chitosan. Response surface methodology based on 3 level Box-Behnken design was applied to analyze the effect of initial pH (4-6), initial concentration (20-60 mg L-1), contact time (15-105 min), and temperature (30-50 °C) on the uptake capacity of uranium(VI). Main effect of initial concentration, quadratic effect of contact time, and dual effect of initial pH and contact time were found statistically significant based on analysis of variance (ANOVA). Probability F-value (F = 1.49 ×10-6) and correlation coefficient (R2 = 0.96) point out that the proposed model is compatible with experimental data. The maximum uptake capacity of uranium(VI) was found as 28.48 mg g-1 at initial pH 4, initial concentration 60 mg L-1, contact time of 70 min, and a temperature of 50 °C. Sorption kinetics followed a pseudo-second-order model and Freundlich model was obtained to fit the sorption data. The presence of competing ions slightly reduced uranium(VI) sorption and the selectivity order can be given as UO2 2+>Zn2+>Ni2+.


Inhibition of [3H]norepinephrine uptake in organ cultured rat superior cervical ganglia by xylamine.
The superior cervical ganglia (SCG) of the rat, when maintained in organ culture, show an increase in [3H]norepinephrine (NE) uptake in the first 2 days and then a leveling off of this activity through 6 days of culture. The [3H]NE uptake in cultured SCG is Na+ - and temperature-dependent and is inhibited by cocaine, desipramine and benztropine with potencies similar to those reported for uptake inhibition into noradrenergic nerve terminals. The preparation also accumulates [3H]dopamine (DA), but, in contrast to NE and DA uptake in the brain, desipramine and benztropine show no specificity for a particular catecholamine, inhibiting both [3H]NE and [3H]DA uptake with potencies similar to those shown for NE uptake in brain. Thus, observed DA uptake and SCG appears to occur mostly via the NE uptake system, not a specific DA uptake system. Xylamine (N-2-chloroethyl-N-ethyl-2-methylbenzylamine) at 10 microM irreversibly inhibits [3H]NE uptake in cultured SCG by 85% in 30 min, uptake being measured after washing away unbound drug. The IC50 for inhibition is about 0.3 microM. When SCG are again cultured after xylamine exposure, [3H]NE uptake recovers after 2 days. This recovery is partially blocked by cycloheximide, indicating that the synthesis of new uptake sites may be involved.


Phase II study of docetaxel, capecitabine, and cisplatin as neoadjuvant chemotherapy for locally advanced breast cancer.
Docetaxel, capecitabine, and cisplatin are effective chemotherapeutic agents for breast cancer with significant synergistic cytotoxicity demonstrated by in vitro studies. The purpose of this study was to assess the efficacy of a combination of docetaxel, capecitabine, and cisplatin (TXP) in patients diagnosed with locally advanced breast cancer (LABC). Patients (n = 42) with chemotherapy-naïve LABC (stage IIIa or IIIb) were enrolled. The chemotherapeutic regimen consisted of 4-6 cycles of intravenous docetaxel (60 mg/m(2)) and cisplatin (50 mg/m(2)) on day 1, plus oral capecitabine (1,800 mg/m(2)/day) on day 1-14, repeated every 3 weeks. Upon completion of therapy, the primary tumor was resected when not contraindicated. Median patient age was 48.5 years (range 31-66 years). Median tumor size was 6.8 cm (range 2.7-15 cm), 29 patients were node-positive, and 12 patients were hormone receptor positive. A total of 216 cycles (median 5; range 3-6 cycles) were administered without prophylactic G-CSF. The predominant toxicities were grade 3/4 neutropenia (30%/28%) and no grade 3/4 thrombocytopenia, febrile neutropenia, or grade 4 non-hematological toxicities were observed. Grade 3 non-hematological toxicities included hand-foot syndrome (5.6%) and vomiting (0.5%). The overall clinical response rate was 97.6% (41/42). Six of the 42 patients (14.3%) achieved a complete pathological response. Of 22 patients who completed 6 cycles of combination treatment, the complete pathological response was 27.3% (6/22). A combination of TXP can be administered safely without prophylactic G-CSF, and appears to be an effective neoadjuvant regimen in patients with LABC.


The CPT1A Arctic variant: perspectives of community members and providers in two Alaska tribal health settings.
Newborn screening in Alaska includes screening for carnitine palmitoyltransferase 1A (CPT1A) deficiency. The CPT1A Arctic variant is a variant highly prevalent among Indigenous peoples in the Arctic. In this study, we sought to elicit Alaska Native (AN) community member and AN-serving healthcare providers' knowledge and perspectives on the CPT1A Arctic variant. Focus groups with community members and healthcare providers were held in two regions of Alaska between October 2018 and January 2019. Thematic analysis was used to identify recurring constructs. Knowledge and understanding about the CPT1A Arctic variant and its health impact varied, and participants were interested in learning more about it. Additional education for healthcare professionals was recommended to improve providers' ability to communicate with family caregivers about the Arctic variant. Engagement with AN community members identified opportunities to improve educational outreach via multiple modalities for providers and caregivers on the Arctic variant, which could help to increase culturally relevant guidance and avoid stigmatization, undue worry, and unnecessary intervention. Education and guidance on the care of infants and children homozygous for the CPT1A Arctic variant could improve care and reduce negative psychosocial effects.


Injuries and overuse syndromes in powerlifting.
Powerlifting is a discipline of competitive weightlifting. To date, no investigations have focused on pain encountered during routine training. The aim of the study was to identify such pain, assign it to particular exercises and assess the data regarding injuries as well as the influence of intrinsic and extrinsic factors. Data of 245 competitive and elite powerlifters was collected by questionnaire. Information regarding current workout routines and retrospective injury data was collected. Study subjects were selected from 97 incorporated powerlifting clubs. A percentage of 43.3% of powerlifters complained of problems during routine workouts. Injury rate was calculated as 0.3 injuries per lifter per year (1 000 h of training=1 injury). There was no evidence that intrinsic or extrinsic factors affected this rate. Most commonly injured body regions were the shoulder, lower back and the knee. The use of weight belts increased the injury rate of the lumbar spine. Rate of injury to the upper extremities was significantly increased based on age >40 years (shoulder/p=0.003, elbow/p=0.003, hand+wrist/p=0.024) and female gender (hand+wrist/p=0.045). The daily workout of a large proportion of powerlifters is affected by disorders which do not require an interruption of training. The injury rate is low compared to other sports.


Incorporation of [2-3H]glycerol into cell surface components of Bacillus subtilis 168 and thermosensitive mutants affected in wall teichoic acid synthesis: effect of tunicamycin.
A method is described for measuring the synthesis of poly(glycerol phosphate) [poly(groP)], the major wall teichoic acid (WTA), lipoteichoic acid (LTA) and phospholipid (P-lipid), through fractionation of [2-3H]glycerol ([2-3H]gro)-labelled Bacillus subtilis cells. When cultures of certain temperature-sensitive mutants defective in one of several tag genes, encoding enzymes involved in WTA synthesis, were transferred to the restrictive temperature, the synthesis of WTA underwent a specific, immediate, block, while that of LTA or P-lipid proceeded unimpeded. These results, in addition to confirming the role of tag genes, demonstrated, reciprocally, the specificity of the fractionation procedure used to distinguish label in WTA from that in LTA or P-lipid. Results of analysis of other, less severely affected, tag-deficient mutants, as well as of another genetically unrelated mutant developing comparable morphological phenotypes in non-permissive conditions, are discussed in relation to a possible mechanism generating the latter phenotype. Fractionation of B. subtilis 168 cells labelled either with [2-3H]gro or with [1-14C]N-acetylglucosamine, to which tunicamycin was added at 0.5 microg ml(-1) (the MIC) revealed a specific and marked inhibition of poly(groP) as well as of poly(3-O-beta-D-glucopyranosyl-N-acetylgalactosamine 1-phosphate), the minor WTA. However, for 60 min at least, the syntheses of PG, LTA and P-lipid were barely affected.


Alteration in propagating colonic contractions by dairy proteins in isolated rat large intestine.
Gastrointestinal conditions in which the transit of contents is altered may benefit from nutritional approaches to influencing health outcomes. Milk proteins modulate the transit of contents along different regions, suggesting that they have varying effects on neuromuscular function to alter gastrointestinal motility. We tested the hypothesis that bovine whey and casein milk protein hydrolysates could have direct modulatory effects on colonic motility patterns in isolated rat large intestine. Casein protein hydrolysate (CPH), whey protein concentrate (WPC), whey protein hydrolysate (WPH), and a milk protein hydrolysate (MPH; a hydrolyzed blend of 60% whey to 40% casein) were compared for their effects on spontaneous contractile waves. These contractions propagate along the length of the isolated intact large intestine (22 cm) between the proximal colon and rectum and were detected by measuring activity at 4 locations. Milk proteins were perfused through the tissue bath, and differences in contraction amplitude and frequency were quantified relative to pretreatment controls. Propagation frequency was decreased by CPH, increased by MPH, and unaffected by intact whey proteins. The reduced motility with CPH and increased motility with MPH indicate a direct action of these milk proteins on colon tissue and provide evidence for differential modulation by hydrolysate type. These findings mirror actions on lower gastrointestinal transit reported in vivo, with the exception of WPH, suggesting that other factors are required.


Bias of nearest neighbor error estimates.
The bias of the finite-sample nearest neighbor (NN) error from its asymptotic value is examined. Expressions are obtained which relate the bias of the NN and 2-NN errors to sample size, dimensionality, metric, and distributions. These expressions isolate the effect of sample size from that of the distributions, giving an explicit relation showing how the bias changes as the sample size is increased. Experimental results are given which suggest that the expressions accurately predict the bias. It is shown that when the dimensionality of the data is high, it may not be possible to estimate the asymptotic error simply by increasing the sample size. A new procedure is suggested to alleviate this problem. This procedure involves measuring the mean NN errors at several sample sizes and using our derived relationship between the bias and the sample size to extrapolate an estimate of the asymptotic NN error. The results are extended to the multiclass problem. The choice of an optimal metric to minimize the bias is also discussed.


Multiple regions contribute to membrane targeting of Rab GTPases.
Small GTPases of the Rab family are key regulators of membrane trafficking. Each Rab shows a characteristic subcellular distribution, and may serve as an important determinant of organelle identity. The molecular mechanisms responsible for targeting Rabs to specific intracellular compartments, however, remain poorly understood. The divergent C-terminal hypervariable region was postulated to contain Rab targeting information. We generated a series of hybrid Rab proteins by exchanging the hypervariable domains of Rab1a, Rab2a, Rab5a, Rab7 and Rab27a, and analysed their subcellular localisations. We found that the various hybrid proteins retained their targeting to the parent organelle and were functionally active. We conclude that the hypervariable region does not contain a general Rab targeting signal. Furthermore, we identified other regions within the RabF and RabSF motifs that are required for specific targeting of Rab27a to secretory granules or melanosomes, and Rab5a to endosomes. We observed only partial overlap between targeting-determining regions in the Rab proteins examined, suggesting that Rab recruitment may be complex and at least partially Rab-specific. Mutations in these targeting-determining regions induced localisation to the ER, an observation that further strengthens our previous finding that ER/Golgi membranes serve as the default location for Rabs that have lost targeting information.


The origin of the central dogma through conflicting multilevel selection.
The central dogma of molecular biology rests on two kinds of asymmetry between genomes and enzymes: informatic asymmetry, where information flows from genomes to enzymes but not from enzymes to genomes; and catalytic asymmetry, where enzymes provide chemical catalysis but genomes do not. How did these asymmetries originate? Here, we show that these asymmetries can spontaneously arise from conflict between selection at the molecular level and selection at the cellular level. We developed a model consisting of a population of protocells, each containing a population of replicating catalytic molecules. The molecules are assumed to face a trade-off between serving as catalysts and serving as templates. This trade-off causes conflicting multilevel selection: serving as catalysts is favoured by selection between protocells, whereas serving as templates is favoured by selection between molecules within protocells. This conflict induces informatic and catalytic symmetry breaking, whereby the molecules differentiate into genomes and enzymes, establishing the central dogma. We show mathematically that the symmetry breaking is caused by a positive feedback between Fisher's reproductive values and the relative impact of selection at different levels. This feedback induces a division of labour between genomes and enzymes, provided variation at the molecular level is sufficiently large relative to variation at the cellular level, a condition that is expected to hinder the evolution of altruism. Taken together, our results suggest that the central dogma is a logical consequence of conflicting multilevel selection.


Head and Neck Cancer Detection in Digitized Whole-Slide Histology Using Convolutional Neural Networks.
Primary management for head and neck cancers, including squamous cell carcinoma (SCC), involves surgical resection with negative cancer margins. Pathologists guide surgeons during these operations by detecting cancer in histology slides made from the excised tissue. In this study, 381 digitized, histological whole-slide images (WSI) from 156 patients with head and neck cancer were used to train, validate, and test an inception-v4 convolutional neural network. The proposed method is able to detect and localize primary head and neck SCC on WSI with an AUC of 0.916 for patients in the SCC testing group and 0.954 for patients in the thyroid carcinoma testing group. Moreover, the proposed method is able to diagnose WSI with cancer versus normal slides with an AUC of 0.944 and 0.995 for the SCC and thyroid carcinoma testing groups, respectively. For comparison, we tested the proposed, diagnostic method on an open-source dataset of WSI from sentinel lymph nodes with breast cancer metastases, CAMELYON 2016, to obtain patch-based cancer localization and slide-level cancer diagnoses. The experimental design yields a robust method with potential to help create a tool to increase efficiency and accuracy of pathologists detecting head and neck cancers in histological images.


[Complexes of histone H1 with supercoiled DNA].
The action of DNA topoisomerase I on the complexes of supercoiled DNA (DNA I) with histone H1 and other histones was studied at different ionic conditions and histone/DNA ratios. In the presence of 0.15 M NaCl at histone H1/DNA ratios lower than 0.25 (w/w) the relaxation of DNA I is not inhibited. Raising of H1/DNA I ratio up to 0.7 is followed by significant inhibition of DNA I relaxation. At fixed H1/DNA I ratio maximal inhibition is detected at 0.25 M NaCl. At NaCl concentrations lower than 0.1 M and higher than 0.3 M increasing of DNA I relaxation in the presence of H1 was observed. Electron microscopic studies show that increase of ionic strength or H1/DNA I ratio causes more dense packing of DNA molecules in the H1.DNA complexes. Changes in the structure of complexes agree with the increase of DNA I relaxation inhibition in these conditions. DNA I relaxation inhibition by H1 is drastically decreased by iodination of tyrosine 72 residue in the globular part of H1 molecule. Individual core histones inhibit DNA I relaxation at much higher histone/DNA ratios and show different dependence of inhibition on ionic strength. The results are discussed in terms of the possible role of H1 in chromatin condensation-decondensation.


[Budgetary impact analysis in health: update with a model using a generic approach].
Budgetary Impact Analysis (BIA) applied to health care can be defined as the estimate of the net financial costs that a given intervention would represent for a health care institution given the case it was covered. Routinely, BIAs are used to decide the inclusion or exclusion of drugs in therapeutic schemes; actually, the increased use of BIAs have raised awareness about the fact that health economic evaluations represent a partial view in the analysis of the consequences of incorporating health technologies. This paper seeks to identify the determinants and components of BIA, and to describe the development of a spreadsheet model that enables us to assess the Budget impact of any health technology and perform estimations with differing degrees of complexity. Its design explicitly adapts to the user skills and gaps in information, thus seeking to promote the development of these tools in the management fields in our countries.


Cu2+-embedded carbon nanoparticles as anticancer agents.
We report the synthesis of luminescent carbon nanoparticles (93 ± 50 nm) embedded with Cu2+. It was observed that at a relatively low concentration of Cu2+ (2.55 ppm), cervical cancer HeLa cells died due to apoptosis induced by the nanoparticles. Also, generation of reactive oxygen species in the cells, in the presence of the composite nanoparticles, has been attributed to their killing. The luminescence of the carbon nanoparticles was used for imaging of the cells.


Cryopreservation of Orchid Pollinia Using the V Cryo-Plate Method.
Preserving pollinia viability and fertility for pollination at specific times of the year is very important in orchid breeding. One possible method is cryopreservation using aluminium cryo-plate which is increasingly used for the long-term storage of plant genetic resources. To determine a V cryo-plate method for the cryopreservation of orchid pollinia and apply it to some Thai orchid species for breeding. Pollinia of Rhynchostylis gigantea (L.) Ridl. were collected from completely open flowers in the morning and then were placed on aluminium cryo-plates embedded in alginate gel, then immersed in loading solution containing 2 M glycerol and 0.4 M sucrose for 15 min at room temperature (29oC), and then dehydrated with PVS2 solution for 0-60 min at 29oC. The cryo-plates with pollinia were directly plunged into liquid nitrogen for 40 min, and rapidly warmed in 1 M sucrose for 15 min. The cryopreserved and non-cryopreserved pollinia were used for hand-pollinating flowers of the same species for producing hybrids. The viability of non-cryopreserved and cryopreserved pollinia dehydrated with PVS2 was 100%. The highest capsule set after pollinating flowers with cryopreserved pollinia dehydrated with PVS2 for 40 min was also up to 100%. The protocol for cryopreservation of R. gigantea (L.) Ridl pollinia using V cryo-plate method was successfully applied for the cryopreservation of nine Thai orchid species. The exposure time to PVS2 affected the pollinia viability (range 40-100 %; average 93%) and capsule set (range 20-100 %; average 78%) of the nine species. The successful capsule set and seed production after pollination with cryopreserved pollinia in all orchid hybrids were observed. Seed germinated into protocorms and developed to plantlets cultured on modified Vacin and Went (1949) agar medium. Cryopreservation of freshly collected orchid pollinia using V cryo-plate method is an efficient tool for the long-term storage of plant germplasm and for orchid breeding.


Influenza vaccine for chronic obstructive pulmonary disease (COPD).
Influenza vaccinations are currently recommended in the care of people with COPD, but these recommendations are based largely on evidence from observational studies, with very few randomised controlled trials (RCTs) reported. Influenza infection causes excess morbidity and mortality in people with COPD, but there is also the potential for influenza vaccination to cause adverse effects, or not to be cost effective. To determine whether influenza vaccination in people with COPD reduces respiratory illness, reduces mortality, is associated with excess adverse events, and is cost effective. We searched the Cochrane Airways Trials Register, two clinical trials registries, and reference lists of articles. A number of drug companies we contacted also provided references. The latest search was carried out in December 2017. RCTs that compared live or inactivated virus vaccines with placebo, either alone or with another vaccine, in people with COPD. Two review authors independently extracted data. All entries were double-checked. We contacted study authors and drug companies for missing information. We used standard methods expected by Cochrane. We included 11 RCTs with 6750 participants, but only six of these included people with COPD (2469 participants). The others were conducted on elderly and high-risk individuals, some of whom had chronic lung disease. Interventions compared with placebo were inactivated virus injections and live attenuated intranasal virus vaccines. Some studies compared intra-muscular inactivated vaccine and intranasal live attenuated vaccine with intra-muscular inactivated vaccine and intranasal placebo. Studies were conducted in the UK, USA and Thailand.Inactivated vaccine reduced the total number of exacerbations per vaccinated participant compared with those who received placebo (mean difference (MD) -0.37, 95% confidence interval (CI) -0.64 to -0.11; P = 0.006; two RCTs, 180 participants; low quality evidence). This was due to the reduction in 'late' exacerbations, occurring after three or four weeks (MD -0.39, 95% CI -0.61 to -0.18; P = 0.0004; two RCTs, 180 participants; low quality evidence). Both in people with COPD, and in older people (only a minority of whom had COPD), there were significantly more local adverse reactions in people who had received the vaccine, but the effects were generally mild and transient.There was no evidence of an effect of intranasal live attenuated virus when this was added to inactivated intramuscular vaccination.Two studies evaluating mortality for influenza vaccine versus placebo were too small to have detected any effect on mortality. However, a large study (N=2215) noted that there was no difference in mortality when adding live attenuated virus to inactivated virus vaccination, AUTHORS' CONCLUSIONS: It appeared, from the limited number of RCTs we were able to include, all of which were more than a decade old, that inactivated vaccine reduced exacerbations in people with COPD. The size of effect was similar to that seen in large observational studies, and was due to a reduction in exacerbations occurring three or more weeks after vaccination, and due to influenza. There was a mild increase in transient local adverse effects with vaccination, but no evidence of an increase in early exacerbations. Addition of live attenuated virus to the inactivated vaccine was not shown to confer additional benefit.


[An experimental model of helicoid ventricular tachycardia (torsades de pointes). Preliminary report].
Studies on twenty dogs were performed. With open thorax, aconitine was injected into the thickness of the interventricular septum. D2 and direct derivations were registered in the epicardium of the right atrium, on right and left septal surfaces and in the His bundle. As the experiment ended, the heart was taken out to determine the placement of the electrodes and the site of application of aconitine. "Torsades de pointes" were obtained in 15 dogs, with a rotation of 180 degrees during the episode. The rotation of A QRS occurred with a beat of intermediate direction which corresponded to a ventricular capture of a sinus beat or a ventricular echo after that beat, and a sequence of activation of AHV established in various beats, the first of which had a prolonged AH (p less than 0.01). The consecutive change of direction of QRS happened when that sequence of activation disappeared, and there was an A-V dissociation as in the beginning of the episode. In conclusion, in this experimental model, the Torsades de pointes were produced by double mechanism, an increased focus of automatism and a macro-re-entry that involves the two branches of the His bundle, the A-V node and the atrium.


Molecular origin of the binding of WWOX tumor suppressor to ErbB4 receptor tyrosine kinase.
The ability of WWOX tumor suppressor to physically associate with the intracellular domain (ICD) of ErbB4 receptor tyrosine kinase is believed to play a central role in downregulating the transcriptional function of the latter. Herein, using various biophysical methods, we show that while the WW1 domain of WWOX binds to PPXY motifs located within the ICD of ErbB4 in a physiologically relevant manner, the WW2 domain does not. Importantly, while the WW1 domain absolutely requires the integrity of the PPXY consensus sequence, nonconsensus residues within and flanking this motif do not appear to be critical for binding. This strongly suggests that the WW1 domain of WWOX is rather promiscuous toward its cellular partners. We also provide evidence that the lack of binding of the WW2 domain of WWOX to PPXY motifs is due to the replacement of a signature tryptophan, lining the hydrophobic ligand binding groove, with tyrosine (Y85). Consistent with this notion, the Y85W substitution within the WW2 domain exquisitely restores its binding to PPXY motifs in a manner akin to the binding of the WW1 domain of WWOX. Of particular significance is the observation that the WW2 domain augments the binding of the WW1 domain to ErbB4, implying that the former serves as a chaperone within the context of the WW1-WW2 tandem module of WWOX in agreement with our findings reported previously. Altogether, our study sheds new light on the molecular basis of an important WW-ligand interaction involved in mediating a plethora of cellular processes.


Sensitivity, specificity, and risk of diagnostic procedures in a canine model of Streptococcus pneumoniae pneumonia.
A canine model in which unilateral Streptococcus pneumoniae pneumonia was induced was used to evaluate the sensitivity, specificity, and risk of various technics proposed for the diagnosis of pulmonary infection. Control and infected animals were studied during spontaneous and mechanical ventilation. We found that three procedures provided a reasonable degree of combined specificity and sensitivity: transthoracic needle aspiration (TTN), catheter-brush biopsy (CBB), and transbronchial biopsy (TBB). Transtracheal aspiration, although sensitive, was consistently low in specificity, whereas TTN provided the highest sensitivity/specificity. Both CBB and TBB provided almost the same results in intubated animals, but specificity was lower in nonintubated animals. Gram-stained smears obtained by TTN, CBB, and TBB were highly specific, but often negative. The only risk encountered was pneumothorax. The TTN was associated with a 20 to 30% risk of pneumothorax; CBB and TBB did not appear to induce pneumothorax. These studies provide the first animal data regarding the comparative yield/risk of these diagnostic approaches. As such, they may prove useful in the design of future human investigations.


Bivalent cholesterol-based coupling of oligonucletides to lipid membrane assemblies.
By mimicking Nature's way of utilizing multivalent interactions, we introduce in the present work a novel method to improve the strength of cholesterol-based DNA coupling to lipid membranes. The bivalent coupling of DNA was accomplished by hybridization between a 15-mer DNA and a 30-mer DNA, being modified with cholesterol in the 3' and 5' end, respectively. Compared with DNA modified with one cholesterol moiety only, the binding strength to lipid membranes appears to be significantly stronger and even irreversible over the time scale investigated ( approximately 1 hr). First, this means that the bivalent coupling can be used to precisely control the number of DNA per lipid-membrane area. Second, the strong coupling is demonstrated to facilitate DNA-hybridization kinetics studies. Third, exchange of DNA between differently DNA-modified vesicles was demonstrated to be significantly reduced. The latter condition was verified via site-selective and sequence-specific sorting of differently DNA-modified lipid vesicles on a low-density cDNA array. This means of spatially control the location of different types of lipid vesicles is likely to find important applications in relation to the rapid progress currently made in the protein chip technology and the emerging need for efficient ways to develop membrane protein arrays.


Cell kinetic basis for pathophysiology of psoriasis.
Studies on the cell proliferation kinetics of psoriatic epidermal cells are presented and the results compared to similar studies for normal epidermis. The short 36-h duration of the psoriatic cell cycle (Tc) is confirmed with the first double-peaked fraction of labeled mitoses (FLM) curve in human subjects. The growth fraction of psoriasis using two experimental techniques approximates 100% within 36 h, confirming the rapid Tc found by the FLM method. The cell kinetic basis for the pathophysiology of psoriasis consists of at least 3 proliferative abnormalities in comparison to normal epidermis. By far the largest alteration is the shortening of the Tc from 311 to 36 h. There is also a doubling of the proliferative cell population in psoriasis from 27,000 to 52,000 cells/mm and an increase in the growth fraction from 60% to 100%. As a consequence of these abnormalities the psoriatic epidermis produces 35,000 cells/day from a proliferative compartment of 52,000 cells/mm2 surface area. This is a 28-fold greater production of cells than the 1,246 cells/day produced in normal epidermis. The biochemical or control factors leading to these kinetic differences continue to remain elusive.


Association of cardiac and renal function with extreme N-terminal fragment pro-B-type natriuretic peptide levels in elderly patients.
The data are inconsistent regarding whether extreme N-terminal fragment pro-B-type natriuretic peptide (NT pro-BNP) levels are associated with impaired renal function. Furthermore, the relationship between extreme NT pro-BNP levels and cardiac and renal function in elderly patients has not been reported. The aim of the present study was to examine a hypothesis that extreme NT pro-BNP levels may be associated with impaired cardiac and renal function in elderly patients. We retrospectively analyzed the data of demographic, clinical, and echocardiographic features on 152 consecutive elderly patients aged more than 80 years old (average age, 83.65 ± 3.58 years) with NT pro-BNP levels ≥ 3000 pg/ml. The participants were divided into two categories according to their NT pro-BNP levels: (1) 3000-10000 pg/mL and (2) >10000 pg /mL. The number of patients with impaired renal function (P = 0.019) and the mortality (P < 0.001) in the period of inpatient was higher in the group with NT pro-BNP > 10000 pg /mL. The levels of serum creatinine and creatine kinase MB (CK-MB) in the group of NT pro-BNP > 10000 pg / mL were higher than those in the group of NT pro-BNP = 3000-10000 pg/mL (P = 0.001 and P = 0.023, respectively). Furthermore, no significant difference in the distribution by NYHA class in different NT pro-BNP levels was observed. Multiple linear regression analyses demonstrated that with NT pro-BNP levels as the dependent variable, NT pro-BNP levels were positively correlated with CK-MB (β = 0.182, P = 0.024) and creatinine levels (β = 0.281, P = 0.001). The area under the receiver-operating characteristic (ROC) curve of NT pro-BNP levels and clinical diagnosis of impaired renal function was 0.596 and reached significant difference (95%CI:0.503-0.688, P = 0.044). These data suggest that the extreme elevation of NT pro-BNP levels (≥3000 pg/ml) is mainly determined by impaired renal function in elderly patients above 80 years. Extreme NT pro-BNP levels may be useful for assessing the severity of impaired renal function.


Effects of regular salmeterol on lung function and exercise capacity in patients with chronic obstructive airways disease.
The aim of this study was to evaluate the effects of single and chronic dosing with salmeterol on exercise capacity and lung function in patients with chronic obstructive pulmonary disease. Twenty nine patients of mean (SE) age 64 (1.5) years, forced expiratory volume in one second (FEV1) 42(3)% of predicted, and 5-15% reversibility to salbutamol 200 micrograms were randomised to receive four weeks treatment with salmeterol 50 micrograms twice daily or placebo in a double blind crossover fashion with a one week washout period in between. Measurements of spirometric parameters, static lung volumes, and exercise capacity were made one and six hours after a single dose, and six hours after the final dose of salmeterol or placebo. Salmeterol produced a small increase in FEV1 at one and six hours after a single dose, and this was maintained after chronic dosing (mean difference and 95% CI versus placebo): single dosing at one hour 0.07 (95% CI 0.02 to 0.11) 1, single dosing at six hours 0.16 (95% CI 0.09 to 0.22) 1, chronic dosing at six hours 0.11 (95% CI 0.03 to 0.19) 1. The increase in forced vital capacity (FVC) was greater with salmeterol than with placebo six hours after single but not chronic dosing: single dosing at six hours 0.17 (95% CI 0.04 to 0.29) 1, chronic dosing at six hours 0.02 (95% CI -0.18 to 0.22) 1. Slow vital capacity was increased after treatment with salmeterol compared with placebo one and six hours after single but not after chronic dosing. There were no significant differences in static lung volumes or exercise capacity after single or chronic dosing with salmeterol compared with placebo. Patients reported a significantly lower Borg score for perceived exertion following the six minute walk after chronic treatment with salmeterol compared with placebo. Salmeterol produced a small improvement in spirometric values compared with placebo consistent with the degree of reversibility originally shown by the subjects to salbutamol 200 micrograms. This was not associated with improvements in static lung volumes or exercise capacity, but there was some symptomatic benefit in that patients were able to walk the same distance in six minutes with less perceived exertion.


The Biological Basis to Personality Disorders.
To provide understanding into the biological basis of thinking and behavior in people with personality disorders, explain anatomic findings, and appraise therapeutic options. PubMed was searched with no date restrictions using the terms personality disorders DSM-5, cluster B personality disorders, biological psychiatry of personality disorders, neurobiology of personality disorders, and neurobiology of cluster B personality disorders. We identified 2,790 English-language articles and utilized 18 in this report. There are anatomic features typical to the brains of individuals with cluster B personality disorders, for example, abnormalities in the superior frontal cortex and amygdala and enlarged striatal volumes. Emotional dysregulation and impulsiveness are 2 prominent symptoms. Hereditary factors may contribute to the development of such conditions. Understanding the neurobiology of cluster B personality disorders expands knowledge that hopefully results in better clinical management and development of improved treatments. Psychotherapy is currently the most effective intervention for borderline personality disorders. Symptomatic pharmacotherapies may be prescribed adjunctively on an individualized basis if clinically indicated (eg, with a coexistant depression).


Three-dimensional hard tissue palatal size and shape: a 10-year longitudinal evaluation in healthy adults.
A 10-year longitudinal evaluation of the morphology (size and shape) of hard tissue palate was performed in 6 female and 6 male healthy adults (mean age at the second evaluation was 33 years, SD = 2.2). All subjects had a complete permanent dentition, including the second molars, and were free from respiratory problems. Palatal landmarks were digitized with a computerized 3D instrument, and their coordinates were used to derive a mathematical model of palatal form. Palatal shape (size-independent) was assessed by a fourth-grade polynomial in the sagittal and frontal plane projections. Palatal dimensions in the frontal and sagittal planes were computed and compared between the 2 evaluations by paired Student t tests. A great variability was observed, and no significant modifications in size were found (P > .05 for all variables). No variations in shape were observed. Sex had no significant effect for any variable (Student t for independent samples, P > .05). This study showed that in healthy subjects, hard tissue palatal morphology does not seem to change between the third and the fourth decades of life.


Chronic Nonbacterial Osteomyelitis: Pathophysiological Concepts and Current Treatment Strategies.
Chronic nonbacterial osteomyelitis (CNO) is an autoinflammatory bone disorder, covering a clinical spectrum with asymptomatic inflammation of single bones at the one end, and chronic recurrent multifocal osteomyelitis (CRMO) at the other end. The exact molecular pathophysiology of CNO remains largely unknown. Provided familial clusters and the association with inflammatory disorders of the skin and intestine suggest a genetic predisposition. Recently, profound dysregulation of cytokine responses was demonstrated in CRMO. Failure to produce antiinflammatory cytokines interleukin (IL)-10 and IL-19 contributes to activation of inflammasomes and subsequent IL-1β release. In IL-10-deficient and in CNO-prone chronic multifocal osteomyelitis mice, IL-1β was linked to bone inflammation. Further, alterations to the gut microbiome were suggested in contributing to IL-1β release from innate immune cells in mice, offering an interesting target in the search for molecular mechanisms in CNO. Here, we summarize clinical presentation and treatment options in CNO/CRMO, current pathophysiological concepts, available mouse models, and promising future scientific directions.


Generic Method for Designing Self-Standing and Dual Porous 3D Bioscaffolds from Cellulosic Nanomaterials for Tissue Engineering Applications.
Three-dimensional scaffolds (3D) with controlled shape, dual porosity and long-term mechanical and dimensional stability in biofluids are of interest as biotemplates in tissue engineering. Herein, self-standing and lightweight cellulose-based biogenic scaffolds with a spatially structured morphology, macropores and interconnected micropores were fabricated using a combination of direct ink writing 3D printing and freeze-drying techniques. This was achieved by developing a water-based and low-cost bicomponent ink based on commercially available nanofibrillated cellulose (NFC) and carboxymethyl cellulose (CMC). Physical cross-linking through dehydrothermal treatment significantly increased the surface hardness, indentation modulus, compression strength, as well as the dimensional stability of the scaffolds in biofluids, in comparison to untreated materials. However, no differences in the spectra of solid state nuclear magnetic resonance or infrared were observed for dehydrothermal treated samples, suggesting that the increase of mechanical properties and dimensional stability is based on the physical cross-linking of functional groups both at the interface between NFC and CMC. The supramolecular structure of the polymers was well-preserved as disclosed by X-ray diffraction measurements. The cross-linked scaffolds showed high proliferation, viability, and attachment of human bone tissue derived osteoblast cells (hFOB). The simple and straightforward avenue proposed here for the design of cellulose-based fibrous inks and dual porous scaffolds from the commercially available materials and without the need of any additional cross-linkers should pave the way for the development of implantable, degradable scaffolds and cell-laden biomaterials for bone tissue regeneration and 3D bioprinting applications.


Predictors of suicidal ideation in people with epilepsy living in Korea.
The risk of suicide or suicide attempts is reported higher in people with epilepsy (PWE) than in the general population. Although epileptic, psychiatric, and psychosocial factors are known risk factors for suicide or suicide attempt, no studies have evaluated the predictors of the severity of suicidal ideation-which is a warning sign for suicide attempts-in PWE. Therefore, we measured the severity of suicidal ideation and its risk factors. Consecutive PWE who were medicated with antiepileptic drugs (AEDs) and attended epilepsy clinic were included in the study. The subjects completed self-reported questionnaires, which included the Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Symptom Checklist-90-Revised (SCL-90-R), and Scale for Suicide Ideation-Beck (SSI-Beck). We compared the patients' demographic and clinical variables, and BDI, BAI, and SCL-90-R scores with their SSI-Beck score, and used our findings to determine the predictors for suicidal ideation. In total, 257 PWE were enrolled in the study. SSI-Beck scores correlated strongly with several seizure-related variables, duration of education, IQ, BDI and BAI scores, and nine domains of the SCL-90-R questionnaire. However, the strongest predictor for suicidal ideation was BDI score (beta=0.41, p<0.001), followed by several SCL-90-R domains, such as obsessive-compulsive (beta=-0.39, p<0.001), depression (beta=0.38, p<0.001), hostility (beta=0.22, p=0.002), paranoid ideation (beta=0.17, p=0.01), and IQ (beta=-0.10, p=0.017). These variables explained 59% of the variance in the SSI-Beck score. The seizure-related variables that influenced the BDI score were seizure frequency, duration of education, MRI abnormality, and number of AEDs. However, these variables explained only 18% of the variance in the BDI score. Major risk factors for suicidal ideation in PWE were depressive and psychiatric symptoms rather than seizure-related variables. Therefore, clinicians should focus on screening for depression and other psychiatric problems and treat them appropriately in order to reduce suicidal behavior in PWE. Since seizure-related variables also exhibited a minor role in determining depressive symptoms, stronger seizure-related risk factors for depression should be sought, such as seizure severity or psychosocial factors, to minimize suicidal behavior.


Urinary cGMP as biological marker of the renal activity of atrial natriuretic factor.
Current evidence suggests guanosine 3',5'-cyclic monophosphate (cGMP) serves as the second messenger for atrial natriuretic factor (ANF) in the kidney in vivo. We examined whether extracellular cGMP accumulation quantitatively reflected the concentration of cGMP within renal cells and whether urinary excretion of cGMP correlated with the physiological action of ANF. cGMP egression was examined in renal epithelial LLC-PK1 cells. ANF augmented intracellular cGMP concentration and extracellular cGMP appearance. Extracellular cGMP was an excellent function of the time-integrated intracellular cGMP concentration. In clearance studies in awake rats, urinary cGMP was primarily of renal cellular origin and correlated with the natriuresis induced by ANF in a time-dependent and concentration-dependent fashion. Urinary cGMP excretion may be useful as a biological marker for the renal activity of ANF in vivo.


Egg handling and storage.
The temperature and relative humidity of storage, as well as the gaseous environment, interact with the fertile egg over time during storage in such a way as to affect the success of incubation either negatively or positively. This interaction occurs both above and below the "physiological zero", at which embryonic metabolism is minimal. This interaction below physiological zero implies that certain physical aspects of the egg must be affected by the environmental conditions. As the eggshell is a relatively fixed component, changes in albumen, shell membranes, cuticle, yolk, or embryo proper must account for these time- and environment-related effects. It is concluded that the major contributor is the albumen, as it is obviously the most dynamic component below physiological zero and is strategically positioned.


Hoarseness as a Predictor for Pulmonary Arterial Aneurysm and Extrinsic Left Main Coronary Artery Compression in Patients with Severe Pulmonary Hypertension.
Pulmonary artery (PA) enlargement is a common finding in patients with severe pulmonary hypertension (PH) and may be associated with extrinsic compression of the left main coronary artery (LMCA-Co) and/or compression of the left recurrent laryngeal nerve resulting in hoarseness named as Ortner syndrome (OS). In this study, we evaluated the diagnostic impact of OS in predicting the PA aneurysm and significant LMCA-Co in patients with PH. Our study population comprised retrospectively evaluated 865 with PH confirmed with the right heart catheterization between 2006 and 2022. Patients underwent coronary angiography due to several indications, including the presence of a PA aneurysm on echocardiography, angina symptoms, or the incidental discovery of LMCA-Co on multidetector computed tomography. The LMCA-Co is defined as diameter stenosis ³ 50% in reference distal LMCA segment on two consecutive angiographic planes. The LMCA-Co and hoarseness were documented in 3.8% and 4.3% of patients with PH, respectively. Increasing PA diameter was significantly associated with worse clinical, hemodynamic, laboratory, and echocardiographic parameters. The receiver operating curves revealed that the PA diameter >41 mm was cutoff for hoarseness (AUC: 0.834; sensitivity 69%, specificity 84%, and negative predictive value 98%), and PA diameter >35 mm was cutoff for LMCA-Co >50% (AUC: 0.794; sensitivity 89%, specificity 58 %, and negative predictive value 99%). An odds ratio of hoarseness for LMCA-Co was 83.3 (95% confidence interval; 36.5-190, P < 0.001) with 3.2% sensitivity, 98.7% specificity, and 59% positive and 98% negative predictive values. In this study, a close relationship was found between the presence of hoarseness and the probability of extrinsic LMCA-Co by enlarged PA in patients with severe PH. Therefore, the risk of LMCA-Co should be taken into account in patients with PH suffering from hoarseness.


A Qualitative Assessment of Barriers and Proposed Interventions to Improve Acute Agitation Management for Children with Mental and Behavioral Health Conditions in the Emergency Department.
Mental health visits to the emergency department (ED) by children are rising in the U.S., and acute agitation during these visits presents safety risks to patients and staff. We sought to assess barriers and strategies for providing high quality care to children who experience acute agitation in the ED. We conducted semi-structured interviews with 6 ED physicians, 6 ED nurses, 6 parents, and 6 adolescents at high risk for developing agitation. We asked participants about experiences with acute agitation care in the ED, barriers and facilitators to providing high quality care, and proposed interventions. Interviews were coded and analyzed thematically. Participants discussed identifying risk factors for acute agitation; worrying about safety and the risk of injury; feeling moral distress; and shifting the culture towards patient-centered, trauma-informed care. Barriers and facilitators included using a standardized care pathway, identifying environmental barriers and allocating resources, partnering with the family and child, and communicating among team members. Nine interventions were proposed: opening a behavioral observation unit with dedicated staff and space, asking screening questions to identify risk of agitation, creating personalized care plans in the electronic health record, using a standardized agitation severity scale, implementing a behavioral response team, providing safe activities and environmental modifications, improving the handoff process, educating staff, and addressing bias and inequities. Understanding barriers can inform solutions to improve care for children who experience acute agitation in the ED. The perspectives of families and patients should be considered when designing interventions to improve care.


Survey of awareness of diabetes mellitus among the Arar population, Northern Border Region of Saudi Arabia.
Diabetes Mellitus (DM) is one of the most widely prevalent diseases in Saudi Arabia. Health education is considered an essential component to improve knowledge and change behavior. People affected by diabetes often have inadequate knowledge about the nature of diabetes, its risk factors and associated complication. The aim of this study was to assess the awareness of the Arar population with various aspects of diabetes mellitus. A cross-sectional study was carried out in Arar city, the capital of the Northern Province of Kingdom of Saudi Arabia (KSA). The study was carried out on Saudi nationals from different age groups that were selected by systematic random sampling. Data was collected by means of personal interview with the participants using a pre-designed questionnaire which was administered by the medical students for each diabetic patient. Data were analyzed by SPSS version 15, using descriptive statistics and Chi-square test. A total of 702 participants were interviewed, among them 201 (28.6%) males and 501 (71.4%) females, and most of them (77.8%) had high educational level, 10.4% were diabetics, 78.9% did not have regular checkup visits to the doctor and 58.5% did not perform any regular exercises, including walking, 60% thought that DM was due to partial or total decrease in insulin secretion and 12.4% thought that it was due to excess sweet eating. Additionally, 48.7% of the respondents thought that lack of exercise and obesity were the major risk factors of DM, 33.2% thought that it was a genetic disease. The majority (86.3) of the participants believed that the treatment of DM was a combination of healthy diet, exercise and medication and more than half (63.1%) said that weight loss and modification of life style were the most important preventive measures of DM. Regarding participants' knowledge about DM complications, 24.5% knew about retinopathy and loss of vision, 8.3% knew about retinopathy, loss of vision, low sensation and numbness in extremities, 24.9% said that symptoms of DM were thirst and frequent urination. This study recommends that health policy makers conduct more effective health education sessions to increase knowledge of diabetic patients and their caregivers about diabetes and the importance of compliance with its treatment.


Challenges and Opportunities for Exploiting the Role of Zeolite Confinements for the Selective Hydrogenation of Acetylene.
Zeolites, with their ordered crystalline porous structure, provide a unique opportunity to confine metal catalysts, whether single atoms (e.g., transition metal ions (TMIs)) or metal clusters, when used as a catalyst support. The confined environment has been shown to provide rate and selectivity enhancement across a variety of reactions via both steric and electronic effects, such as size exclusion and transition state stabilization. In this review, we provide a survey of various zeolite confined catalysts used for the semihydrogenation of acetylene highlighting their performance, defined by ethylene selectivity at full acetylene conversion, in relationship to the synthesis technique employed. Synthesis methods that ensure confinement with the catalyst transition metal location in the extra-framework positions are reported to have the highest selectivity to ethylene. However, the underlying molecular factors responsible for selective catalysis within confinement remain elusive due to the difficulty in deconvoluting individual effects. Through the careful use of a combination of characterization and spectroscopic methods, insights into the relationship between the properties of zeolite confined catalysts and their performance have been explored in other works for a variety of reactions. More specifically, operando spectroscopy studies have revealed the dynamic behavior of zeolite confined catalysts under various conditions implying that the structure and properties observed ex situ do not always match those of the active catalyst under reaction conditions. Applying this type of analysis to acetylene semihydrogenation, a simple gas phase reaction, can help elucidate the structure-function relationship of zeolite confined catalysts allowing for more informed design choices and consequently their application to a wider variety of more complex reactions such as the liquid phase hydrogenation of alkynols where solvent effects must also be considered in addition to those of confinement.


Determinants of Doppler-based renal resistive index in patients with septic shock: impact of hemodynamic parameters, acute kidney injury and predisposing factors.
Increased renal resistive index (RI) measured by Doppler ultrasonography has been shown to be associated with acute kidney injury (AKI) in septic patients. However, its clinical use is limited by poor sensitivity and specificity which may be explained by its numerous determinants [in particular mean arterial pressure (MAP)]. We measured, in patients with septic shock, RI at different MAP levels over a short period of time on the admission day to ICU (D1) and every 3 days until day 10 (D10) to define the determinants of RI and study specifically the relationship between RI and MAP. Consecutive patients with septic shock without preexisting chronic renal dysfunction were included in this prospective cohort study in two ICUs. Sixty-five patients were included in the study. Thirty-three (50.8%) and 15 (23.1%) patients had a history of chronic hypertension or diabetes, respectively. At D3, 35 patients presented AKI with AKIN 2 or 3 criteria (severe AKI, AKIN2-3 group) and 30 presented no AKIN or AKIN 1 criteria (AKIN0-1 group). As previously described, RI at D1 was higher in the AKIN2-3 group than in the AKIN0-1 group (0.73 interquartile range [0.67; 0.78] vs. 0.67 [0.59; 0.72], p = 0.001). A linear mixed model for predicting RI from D1 to D10 showed that an increase in pulse pressure, presence of severe AKI and additional day of ICU hospitalization were associated with an increase in RI. An increase in MAP and recovery from severe AKI were associated with a decrease in RI. In the presence of chronic hypertension or diabetes, an increase in MAP resulted in a lower decrease in RI, than in the absence of such factors. Presence of AKI at D3 did not impact the relationship between MAP and RI. Severe AKI was associated with a reversible increase in RI without significant interaction with the relationship between MAP and RI. Conversely, the presence of chronic hypertension and/or diabetes interacted with this relationship.


Developmental exposure to the organophosphorus flame retardant tris(1,3-dichloro-2-propyl) phosphate: estrogenic activity, endocrine disruption and reproductive effects on zebrafish.
Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) is an organophosphate flame retardant that is detectable in the environment and biota, prompting concern over its risk to wildlife and human health. Our objective was to investigate whether long-term exposure to low concentrations of TDCPP can affect fish reproduction. Zebrafish embryos were exposed to low concentrations (0, 4, 20 and 100μg/L) of TDCPP from 2h post-fertilization until sexual maturation. Exposure to TDCPP significantly increased plasma estradiol and testosterone levels in females, but had no effect in males. TDCPP exposure also caused a significant reduction in fecundity as indicated by decreased egg production. Real-time PCR was performed to examine selected genes in the hypothalamic-pituitary-gonadal (HPG) axis and liver. Principle component analysis (PCA) showed that sex hormone levels and fecundity were related to the mRNA level of several genes in the HPG axis. Furthermore, hepatic vitellogenin (vtg1 and vtg3) expression was upregulated in both females and males, suggesting TDCPP has estrogenic activity. Histological examination revealed promotion of oocyte maturation in the females, but retardation of spermiation in males. Reduced egg quality (e.g., egg diameter) and increased malformation rates were observed in the F1 generation. Chemical analysis showed significant levels of TDCPP and its metabolite bis(1,3-dichloro-2-propyl) phosphate in the gonads of males and females. In conclusion, long-term exposure to low concentrations of TDCPP impairs fish reproduction.


How to … define clinical education research terminology: A glossary.
Clinical education research (ClinEdR) utilises diverse terminology, which can lead to confusion. A common language is essential for enhancing impact. An expert panel drawn from various workstreams within the National Institute for Health and Care Research (NIHR) Incubator for Clinical Education Research was tasked with reviewing an initial list of terms for the development of a glossary of terms in the field of ClinEdR. The glossary was populated with terms, definitions and foundational papers by the authors and peer-reviewed for accuracy. The glossary of terms developed for ClinEdR should enable researchers to use a common language, promoting consistency and improving communication. We anticipate this will be useful for ClinEdR students and early career researchers. The glossary could be integrated into educational research methods courses in ClinEdR, and through critical and reflective use, enhance the quality and subsequent impact of ClinEdR.


The analysis of sex segregation: when is index measurement not index measurement?
In their paper in this issue, Grusky and Charles (1998) make a number of dubious claims about the measurement and interpretation of sex segregation. First, they incorrectly claim that only log-odds measures yield margin-free measures of segregation. Second, the estimation and testing of a limited class of log-linear models does not provide an independent test of the appropriateness of a log-odds ratio index to measure segregation. Their estimation in forms them of the statistically justifiable degree of occupational disaggregation, not whether a log-odds ratio is superior to, say, a linear index in the measurement of segregation. Finally, their index A is beset with problems of interpretation, not withstanding their arguments, and their additional measures, AW and AB, suffer similar problems. Grusky and Charles are, however, correct in arguing that measurement procedures should be margin-free. Further, I concur with the view that the adoption of a single annual summary measure of segregation cannot be justified, because it is premised on the assumption that individual occupations, or groups of occupations, exhibit similar trends in sex segregation.


Emerging new sites for adult neurogenesis in the mammalian brain: a comparative study between the hypothalamus and the classical neurogenic zones.
In adult mammalian brain, two main germinative regions located in the subventricular zone of the lateral ventricle and in the subgranular cell layer of the hippocampal dentate gyrus have been considerably documented and are still under intense scrutiny. However, new neuron formation has recently been reported in various other brain areas including the hypothalamus. This central structure, responsible for the control of many major neuroendocrine functions such as reproduction, expresses high levels of PSA-NCAM and nestin, both proteins being involved in structural and morphological plasticity mechanisms. Cell proliferation and new neuron production have been demonstrated in the adult hypothalamus of numerous species, although not hitherto described in non-human primates and humans. Similarly to the subventricular zone and in the subgranular cell layer, the adult hypothalamic neurogenesis process is subject to dynamic regulation by various physiological and pharmacological signals. Several pieces of evidence support the hypothesis that a stem cell niche-like architecture exist in the hypothalamus region lining the third ventricle thereby enabling adult neural stem cells to continuously generate neurons in vivo throughout life. Furthermore, recent data indicating that new hypothalamic neurons may become functionally implicated in sensory information processing endorse the assumption that the hypothalamus might be a neurogenic region.


Supplemental carbohydrate sources for lactating dairy cows on pasture.
Two experiments were conducted to evaluate steam-flaked corn and nonforage fiber sources as supplemental carbohydrates for lactating dairy cows on pasture. Cows were allotted to a new paddock of an orchardgrass (Dactylis glomerata L.) pasture twice daily in one group in both trials. In experiment 1, 28 Holstein cows, averaging 216 d in milk, were randomly assigned to either a cracked-corn (CC) or a steam-flaked (SFC) supplement in a split plot design. The supplement contained 66.7% of corn and a protein/mineral pellet. In experiment 2, 28 Holstein cows, averaging 182 d in milk, were randomly assigned to either a ground corn (GC) or a nonforage fiber (NFF)-based supplemented in a single reversal design. The GC supplement contained 85% ground corn plus protein, mineral, and vitamins. The NFF supplement contained 35% ground corn, 18% beet pulp, 18% soyhulls, 8% wheat middlings plus protein, mineral, and vitamins. In both experiments, cows were fed the grain supplement twice daily after each milking at 1 kg/4 kg milk. In experiment 1, milk production (24.3 kg/d) and composition did not differ between treatments; however, plasma and milk urea N were lower with the SFC supplement. In experiment 2, milk production (27.5 kg/d) was not affected by treatments, which may be related to the medium quality of pasture grazed. The GC supplement tended to reduce plasma and milk urea N and increased milk protein percentage (3.23 vs. 3.19%). Pasture dry matter intake, measured using Cr2O3, did not differ between treatments in either experiment 1 (15.1 kg/d) or experiment 2 (12.2 kg/d). Milk production did not differ when mid-late lactation cows on pasture were supplemented with SFC or NFF instead of dry corn.


The Effect of Selective Laser Melting Process Parameters on the Microstructure and Mechanical Properties of Al6061 and AlSi10Mg Alloys.
Additive manufacturing (AM) offers customization of the microstructures and mechanical properties of fabricated components according to the material selected and process parameters applied. Selective laser melting (SLM) is a commonly-used technique for processing high strength aluminum alloys. The selection of SLM process parameters could control the microstructure of parts and their mechanical properties. However, the process parameters limit and defects obtained inside the as-built parts present obstacles to customized part production. This study investigates the influence of SLM process parameters on the quality of as-built Al6061 and AlSi10Mg parts according to the mutual connection between the microstructure characteristics and mechanical properties. The microstructure of both materials was characterized for different parts processed over a wide range of SLM process parameters. The optimized SLM parameters were investigated to eliminate internal microstructure defects. The behavior of the mechanical properties of parts was presented through regression models generated from the design of experiment (DOE) analysis for the results of hardness, ultimate tensile strength, and yield strength. A comparison between the results obtained and those reported in the literature is presented to illustrate the influence of process parameters, build environment, and powder characteristics on the quality of parts produced. The results obtained from this study could help to customize the part's quality by satisfying their design requirements in addition to reducing as-built defects which, in turn, would reduce the amount of the post-processing needed.


Benign fibro-osseous lesions of the maxillas: analysis of 11 cases.
A study is made of the principal characteristics of the oral lesions biopsied in our Service of Oral Surgery and histologically diagnosed as corresponding to fibro-osseous lesions of the maxillas. A retrospective review was made of all the biopsies made in a Service of Oral Surgery between 1996 and 2003. The reason for consultation was analyzed, along with patient age, sex, clinical and radiological characteristics, tentative diagnosis, histological diagnosis and treatment provided. A total of 1238 biopsies were performed during the study period. Of these, only 11 corresponded to benign fibro-osseous lesions (7 women and 4 men). The mean patient age was 44 years (range 19-72 years). The most frequent location was the mandible (8 of the cases). In 7 patients the lesions constituted casual radiological findings; 4 presented bulging of the vestibular cortical bone, though only one of them reported pain. The histological diagnoses comprised 7 cemento-ossifying fibromas and 4 fibrous dysplasias. In 9 cases surgical resection was carried out, while in one case an incisional biopsy was performed, and in the remaining case curettage was decided. These lesions are more frequent in women than in men, and the age at presentation is variable. In terms of lesion location, fibrous dysplasia is more common in the upper maxilla, while cemento-ossifying fibroma is more frequently found in the mandible. The diagnosis of such lesions is established upon contrasting the data obtained from the anamnesis, physical examination, the radiological characteristics, the intraoperative findings and the histological study, since both disorders have similar clinical and histological features--despite the fact that they constitute distinct disease conditions.


Impact of rapid, culture-independent diagnosis of candidaemia and invasive candidiasis in a community health system.
Delay in treatment of candidaemia and invasive candidiasis remains a cause of significant morbidity and mortality in high-risk patients. Widespread empirical utilization of antifungal therapy often occurs in an effort to minimize this risk. This study assessed the impact of the T2Candida Panel in a multi-hospital community health system on time to initiation of antifungal therapy in candidaemic patients as well as the utilization of micafungin. Outcomes were compared between those patients with candidaemia prior to T2Candida implementation and those after implementation. Micafungin utilization for patients with suspected candidaemia/invasive candidiasis was compared with that for patients with a negative T2Candida Panel post-implementation. There was a significant decrease in time to appropriate therapy in the post-T2Candida group (34 versus 6 h, P = 0.0147). Empirical antifungal therapy was avoided in 58.4% of T2Candida-negative patients. These results support the implementation of T2Candida to improve time to appropriate therapy for candidaemic patients while simultaneously expanding antimicrobial stewardship efforts to appropriately utilize antifungals.


Whole Blood in Pediatric Craniofacial Reconstruction Surgery.
Pediatric complex cranial vault reconstruction (CCVR) surgery is often associated with significant blood loss and transfusion. The authors recently changed our transfusion practice during CCVR to using whole blood (WB) instead of reconstituted blood (RB). The aim of this study was to assess the impact of this practice change. Our hypothesis was that replacement with WB would be as effective as RB for the outcomes of total perioperative blood donor exposures (BDEs) and the incidence of laboratory evidence of postoperative coagulopathy. The authors queried the Pediatric Craniofacial Surgery Perioperative Registry for children ages ≤48 months from our institution who underwent CCVR and received either RB or WB. The primary outcomes of total perioperative BDEs and the incidence of laboratory evidence of postoperative coagulopathy were compared between the 2 cohorts. The query returned 59 subjects in the RB cohort and 52 subjects in the WB cohort. There were no significant differences in demographic variables between the 2 groups. Patients in the WB cohort were more likely to have ≤1 BDEs when compared to the RB cohort (62% versus 39%, respectively, P = 0.02). There was no significant difference in the incidence of postoperative coagulation laboratory test abnormalities between the WB and RB cohorts (0% versus 3.4%, respectively, P = 0.50). There was no postoperative coagulopathy in the WB cohort. Whole blood was also associated with significantly fewer perioperative BDEs. Whole blood appears to be as effective as RB for replacement of blood loss in craniofacial surgery.


The effect of green tea supplementation on the anthropometric outcomes in overweight and obese women: a time and dose-response meta-analysis of randomized controlled trials.
Inconsistencies are evident within the literature regarding the role of Green Tea (GT) supplementation on women living with obesity. To address this, we conducted to determine the impact of GT supplementation on the weight, body mass index (BMI), and waist circumference (WC) in overweight and obese women using time and dose-response meta-analysis of randomized controlled trials (RCTs). This meta-analysis searched electronic Scopus, Web of Science, Embase, and PubMed/Medline databases from inception to December 1st, 2022. Data were reported as weighted mean difference (WMD) with 95% confidence interval (CI). A total of 2061 references were identified, and 15 articles with 16 RCT arms on body weight, 17 RCT arms on BMI, and 7 RCT arms on WC were included in the meta-analysis. GT supplementation significantly decreases body weight (WMD: -1.23 kg, 95% CI: -2.13 to -0.33, p = 0.007), BMI (WMD: -0.47 kg/m2, 95% CI: -0.87 to -0.07, p = 0.020) and WC (WMD: -3.46 cm, 95% CI: -6.75 to -0.16, p = 0.040). In subgroup analyses, GT consumption demonstrated lowered body weight with dosaes ≥1000 mg/day (WMD: -1.38 kg), in the RCTs, which lasted ≥8 wk (WMD: -1.24 kg). The non-linear dose-response assessment detected a negative correlation between the changes in body weight and BMI in green tea consumption of more than 1000 (mg/day). The GT supplementation reduced the weight, BMI, and WC in overweight and obese women. In clinical practice, healthcare professionals can recommend using GT with dosages ≥ 1000mg/day and duration ≥ 8 wk in obese women.


Monensin blocks the transport of diphtheria toxin to the cell cytoplasm.
Lysosomotropic amines are believed to inhibit the transport of diphtheria toxin to the cell cytoplasm by raising the pH within intracellular vesicles. If so, then other drugs that dissipate intracellular proton gradients should have a similar effect on toxin transport. We found that monensin, a proton ionophore unrelated to lysosomotropic amines, is a potent inhibitor of the cytotoxic effect of diphtheria toxin. Monensin appears to block the escape of endocytosed toxin from a vesicle to the cytoplasm. Monensin fails to protect cells from the effects of diphtheria toxin that is bound to the cell surface and exposed to acidic medium, suggesting that the step normally blocked by the drug is circumvented under these conditions. The inhibition of toxin transport caused by monensin could not be relieved when monensin was replaced by ammonium chloride, nor when ammonium chloride was again replaced by monensin. This suggests that both drugs block the same step of toxin transport. The effect of monensin on the transport of diphtheria toxin to the cytoplasm is consistent with the proposal (Draper and Simon. 1980. J. Cell Biol. 87:849-854; Sandvig and Olsnes. 1980. J. Cell Biol. 87:828-832) that the toxin is endocytosed and then, in response to an acidic environment, penetrates through the membrane of an intracellular vesicle to reach the cytoplasm.


Nutriepigenetics and cardiovascular disease.
We present a current perspective of epigenetic alterations that can lead to cardiovascular disease (CVD) and the potential of dietary factors to counteract their actions. In addition, we discuss the challenges and opportunities of dietary treatments as epigenetic modifiers for disease prevention and therapy. Recent epigenome-wide association studies along with candidate gene approaches and functional studies in cell culture and animal models have delineated mechanisms through which nutrients, food compounds and dietary patterns may affect the epigenome. Several risk factors for CVD, including adiposity, inflammation and oxidative stress, have been associated with changes in histone acetylation, lower global DNA methylation levels and shorter telomere length. A surplus of macronutrients such as in a high-fat diet or deficiencies of specific nutrients such as folate and other B-vitamins can affect the activity of DNA methyltransferases and histone-modifying enzymes, affecting foetal growth, glucose/lipid metabolism, oxidative stress, inflammation and atherosclerosis. Bioactive compounds such as polyphenols (resveratrol, curcumin) or epigallocatechin may activate deacetylases Sirtuins (SIRTs), histone deacetylases or acetyltransferases and in turn the response of inflammatory mediators. Adherence to cardioprotective dietary patterns, such as the Mediterranean diet (MedDiet), has been associated with altered methylation and expression of genes related to inflammation and immuno-competence. The mechanisms through which nutrients and dietary patterns may alter the cardiovascular epigenome remain elusive. The research challenge is to determine which of these nutriepigenetic effects are reversible, so that novel findings translate into effective dietary interventions to prevent CVD or its progression.


Exercise Oncology and Immuno-Oncology; A (Future) Dynamic Duo.
Recent advances in clinical oncology is based on exploiting the capacity of the immune system to combat cancer: immuno-oncology. Thus, immunotherapy of cancer is now used to treat a variety of malignant diseases. A striking feature is that even patients with late-stage disease may experience curative responses. However, most patients still succumb to disease, and do not benefit from treatment. Exercise has gained attention in clinical oncology and has been used for many years to improve quality of life, as well as to counteract chemotherapy-related complications. However, more recently, exercise has garnered interest, largely due to data from animal studies suggesting a striking therapeutic effect in preclinical cancer models; an effect largely mediated by the immune system. In humans, physical activity is associated with a lower risk for a variety of malignancies, and some data suggest a positive clinical effect for cancer patients. Exercise leads to mobilization of cells of the immune system, resulting in redistribution to different body compartments, and in preclinical models, exercise has been shown to lead to immunological changes in the tumor microenvironment. This suggests that exercise and immunotherapy could have a synergistic effect if combined.


Progressive aggregation of alpha-synuclein and selective degeneration of lewy inclusion-bearing neurons in a mouse model of parkinsonism.
Aggregated alpha-synuclein inclusions are found where cell death occurs in several diseases, including Parkinson's disease, dementia with Lewy bodies, and multiple-system atrophy. However, the relationship between inclusion formation and an individual cell's fate has been difficult to study with conventional techniques. We developed a system that allows for in vivo imaging of the same neurons over months. We show that intracerebral injection of preformed fibrils of recombinant alpha-synuclein can seed aggregation of transgenically expressed and endogenous alpha-synuclein in neurons. Somatic inclusions undergo a stage-like maturation, with progressive compaction coinciding with decreased soluble somatic and nuclear alpha-synuclein. Mature inclusions bear the post-translational hallmarks of human Lewy pathology. Long-term imaging of inclusion-bearing neurons and neighboring neurons without inclusions demonstrates selective degeneration of inclusion-bearing cells. Our results indicate that inclusion formation is tightly correlated with cellular toxicity and that seeding may be a pathologically relevant mechanism of progressive neurodegeneration in many synucleinopathies.


Crohn's disease affecting the nasal mucosa.
Crohn's disease is a chronic inflammatory disease of the gastrointestinal tract, associated with disorders of cell mediated and humoral immunity. The incidence of the disease is increasing, and it is commonest in young women. Any part of the gastrointestinal tract may be affected, and 9% of cases have oral lesions which are usually painful and coincide with periods of active intestinal disease. These oral lesions are areas of cobblestoned mucosa with deep ulcers and fissures; on histological examination there is perivascular inflammation with lymphocytic infiltration and in 10% non-caseating granulomas are seen. In the absence of any clinically detectable lesion, biopsies of buccal mucosa from patients with Crohn's disease show focal inflammation around salivary glands, and immunofluorescent studies have demonstrated antibodies to buccal mucosa in their serum. Involvement of the lower respiratory tract by Crohn's disease is rare, and although laryngeal involvement leading to upper airway obstruction has been described, involvement of the nose has not been previously reported.


Declining inequality? The changing impact of socio-economic background and ability on education in Australia.
The paper addresses several debates surrounding the reproduction of socio-economic inequality: (i) the persistent inequality thesis, which maintains that despite the increases in educational participation socio-economic inequalities in education have not declined; (ii) the related thesis of maximally maintained inequality, which proposes that socio-economic inequalities decline only when participation levels for the most privileged socio-economic group approach saturation levels; (iii) the meritocracy debate on the importance of ability vis-à-vis socio-economic background and changes in its influence over time; and (iv) the effect of policy changes on socio-economic inequalities in education. These issues are addressed using data from six Australian youth cohorts born between 1961 and the mid-1980s.


Exploration of hitherto-uncultured bacteria from the rhizosphere.
The rhizosphere environment selects a particular microbial community that arises from the one present in bulk soil due to the release of particular compounds in exudates and different opportunities for microbial colonization. During plant-microorganism coevolution, microbial functions supporting plant health and productivity have developed, of which most are described in cultured plant-associated bacteria. This review discusses the state of the art concerning the ecology of the hitherto-uncultured bacteria of the rhizosphere environment, focusing on Acidobacteria, Verrucomicrobia and Planctomycetes. Furthermore, a strategy is proposed to recover bacterial isolates from these taxa from the rhizosphere environment.


Gene-for-gene interactions between Pseudomonas syringae pv. phaseolicola and Phaseolus.
The gene for cultivar-specific avirulence to Phaseolus vulgaris cv. Tendergreen in races 3 and 4 of Pseudomonas syringae pv. phaseolicola was isolated and sequenced. Genomic clones from libraries of race 3 in pLAFR1 and race 4 in pLAFR3, which altered the phenotype of race 5 from virulent to avirulent in Tendergreen, were found to possess a common approximately 15-kb region of DNA that contained the determinant of avirulence. Subcloning and insertion mutagenesis with Tn1000 located an avirulence gene within a 1.4-kb BglII/HindIII DNA fragment in races 3 and 4. Comparison of the nucleotide sequences of regions of DNA that confer avirulence confirmed that both races have an identical gene for avirulence (designated avrPph3) comprising 801 base pairs (bp) and predicted to encode a cytoplasmic protein of 28,703 Da. A sequence, TGCAACCGAAT, 91% homologous to the motif found in promoter regions of avrB and avrD from P. s. pv. glycinea was located 89-99 bp upstream of the start of the open-reading frame 1. Hybridization experiments showed that avrPph3 was not plasmid-borne and was absent from isolates of P. s. pv. phaseolicola races 1, 2, 5, 6, 7, and 8, P. cichorii, P. s. pvs. coronafaciens, glycinea, maculicola, pisi, syringae, and tabaci. Cosegregation studies of crosses between cultivars resistant (Tendergreen) and susceptible (Canadian Wonder) to races 3 and 4 and transconjugants of race 5 confirmed that a gene-for-gene relationship controls specificity in the interaction between Tendergreen and races 3 and 4 of P. s. pv. phaseolicola.


Circa- and ultradians in the occurrence of simple extrasystoles in healthy men at lowland in the light of inferential statistics.
An exact description of circadian and ultradian rhythms of simple extrasystoles in healthy subjects is lacking. Thirty-seven healthy male subjects, aged 50-76, were randomly taken. Simple extrasystoles from 24-hour Holter ECG were registered and calculated per each hour and processed by cosinor regression. Their occurrence is formulated by 95% confidence and tolerance corridors for supraventricular and ventricular extrasystoles. The number of extrasystoles is relatively low, dispersion of ventricular extrasystoles is significantly higher than supraventricular ones. A significant increase of their frequency versus the general zero trend straight line was found only around 9 a.m. In supraventricular ones a significant increase around 6-7 a.m., 9 a.m., noon, and 4-5 p.m. and a significant depression around 1-2 a.m. and 10 p.m. was present. Only the 24-hour rhythm is significantly present (alpha=0.05) in ventricular beats while in supraventricular ones also the period lengths of 12, 6, 4, 3.4 and 2.4 hours are significant. The significant difference between supraventricular and ventricular extrasystoles exists in the 24-hour amplitudes and 6-hour acrophases. One circadian and several ultradian rhythms of simple extrasystoles are present in healthy male subjects over 50 years of age. The 95% tolerance chronograms can be exploited in clinical practice.


An extracorporeal perfusion system to bridge pediatric liver transplant candidates.
An extracorporeal liver perfusion system was designed to maintain cadaver livers in an oxygenated, normothermic state for bridging procedures for hepatic transplantation. Nonpulsatile high flow and pulsatile low flow blood are supplied to the portal venous (PV) and hepatic arterial (HA) circulations. Controlled low blood flow (5-10 cc/kg [patient]/min) is exchanged between the high flow extracorporeal perfusion circuit (1 cc/g [liver]/min) and the patient. The system was evaluated in perfusions of fresh, excised pig livers (n = 5). The average oxygen consumption was 9 +/- 3 microliters/g/min, and bile production averaged 4.7 microliters/g/hr. Perfusion pressures and flows were normal in both the HA and PV circulations for about 4 hr. Pressures then gradually rose, especially in the HA circulation, causing flow to decrease, with subsequent mottling and discoloration of the liver. Red blood cell, platelet, and white blood cell counts fell continuously. Maintenance of liver function was assessed by clearance of an 80 mg taurocholic acid challenge. An average of 56% of injected acid was cleared from the perfused livers (n = 5) in the first half hour, compared with 90% and 25% for the in situ (n = 3) and unperfused (n = 3) control livers, respectively. The system consistently maintained livers in a moderately well functioning state through the first 4 hr of perfusion. Adequate support of animals with induced hepatic failure must now be demonstrated.


Construction of pancreas-muscle-liver microphysiological system (MPS) for reproducing glucose metabolism.
Although in vitro models are widely accepted experimental platforms, their physiological relevance is often severely limited. The limitation of current in vitro models is strongly manifested in case of diseases where multiple organs are involved, such as diabetes and metabolic syndrome. Microphysiological systems (MPS), also known as organ-on-a-chip technology, enable a closer approximation of the human organs and tissues, by recreating the tissue microenvironment. Multiorgan MPS, also known as multiorgan-on-a-chip or body-on-a-chip, offer the possibility of reproducing interactions between organs by connecting different organ modules. Here, we designed a three-organ MPS consisting of pancreas, muscle, and liver, to recapitulate glucose metabolism and homeostasis by constructing a mathematical model of glucose metabolism, based on experimental measurement of glucose uptake by muscle cells and insulin secretion by pancreas cells. A mathematical model was used to modify the MPS to improve the physiological relevance, and by adding the liver model in the mathematical model, physiological realistic glucose and insulin profiles were obtained. Our study may provide a methodological framework for developing multiorgan MPS for recapitulating the complex interaction between multiple organs.


Meta-analysis of the relative efficiency of methionine-hydroxy-analogue-free-acid compared with DL-methionine in broilers using nonlinear mixed models.
The purpose of this paper was to perform a meta-analysis to compare the biological efficiency of DL-methionine with methionine-hydroxy-analogue-free-acid in broiler chickens. A database was developed which contained dose-response studies of these 2 methionine sources. Criteria for entry into the database were defined before the meta-analysis was initiated. Data from 46 dose-response experiments, extracted from a total of 27 peer-reviewed papers, were used for the analysis with the Statistical Analysis System. Initially, the NLIN procedure was applied to fit an exponential model of the form y = alpha + beta*[1 - exp (-Gamma*dose)] + e. Thereafter, meta-analysis was conducted by means of nonlinear mixed models, which were fitted by a full maximum likelihood method as implemented in the NLMIXED procedure. The nonlinear model used for the analysis allowed for separate plateaus or different efficiencies of the methionine sources. Mixed modeling was applied to account for heterogeneity among the studies in all regression parameters for both sources of methionine via random effects. Statistical hypotheses were tested by the asymptotic Wald test. In addition, potential co-variables were tested for inclusion as linear regressors for the nonlinear model parameters. In conclusion, the null hypothesis of equal plateaus of the 2 methionine sources was not rejected. The analyses of the response variables average daily gain (ADG) and gain to feed ratio (GF) showed a highly significant difference between the tested methionine sources. The covariate age at start of experiment significantly affected the intercept term alpha for the response variables ADG and GF, respectively. The meta-analysis showed that biological efficiencies of DL-methionine-hydroxy-analogue-free-acid were 81 and 79% of the values for DL-methionine, on an equimolar basis, for ADG and GF, respectively.


Isolation and characterization of Alicycliphilus denitrificans strain BC, which grows on benzene with chlorate as the electron acceptor.
A bacterium, strain BC, was isolated from a benzene-degrading chlorate-reducing enrichment culture. Strain BC degrades benzene in conjunction with chlorate reduction. Cells of strain BC are short rods that are 0.6 microm wide and 1 to 2 microm long, are motile, and stain gram negative. Strain BC grows on benzene and some other aromatic compounds with oxygen or in the absence of oxygen with chlorate as the electron acceptor. Strain BC is a denitrifying bacterium, but it is not able to grow on benzene with nitrate. The closest cultured relative is Alicycliphilus denitrificans type strain K601, a cyclohexanol-degrading nitrate-reducing betaproteobacterium. Chlorate reductase (0.4 U/mg protein) and chlorite dismutase (5.7 U/mg protein) activities in cell extracts of strain BC were determined. Gene sequences encoding a known chlorite dismutase (cld) were not detected in strain BC by using the PCR primers described in previous studies. As physiological and biochemical data indicated that there was oxygenation of benzene during growth with chlorate, a strategy was developed to detect genes encoding monooxygenase and dioxygenase enzymes potentially involved in benzene degradation in strain BC. Using primer sets designed to amplify members of distinct evolutionary branches in the catabolic families involved in benzene biodegradation, two oxygenase genes putatively encoding the enzymes performing the initial successive monooxygenations (BC-BMOa) and the cleavage of catechol (BC-C23O) were detected. Our findings suggest that oxygen formed by dismutation of chlorite can be used to attack organic molecules by means of oxygenases, as exemplified with benzene. Thus, aerobic pathways can be employed under conditions in which no external oxygen is supplied.