Intake of dietary plant sterols is inversely related to serum cholesterol concentration in men and women in the EPIC Norfolk population: a cross-sectional study.
We examined the relation between intake of natural dietary plant sterols and serum lipid concentrations in a free-living population. Cross-sectional population-based study of 22,256 men and women aged 39-79 y resident in Norfolk, UK, participating in the European Prospective Investigation into Cancer (EPIC-Norfolk). MAIN EXPOSURE AND OUTCOME MEASURES: Plant sterol intake from foods and concentrations of blood lipids. Mean concentrations of total cholesterol and low-density lipoprotein cholesterol, adjusted for age, body mass index and total energy intake, decreased with increasing plant sterol intake in men and women. Mean total serum cholesterol concentration for men in the highest fifth of plant sterol intake (mean intake 463 mg daily) was 0.25 mmol/l lower and for low-density lipoprotein cholesterol 0.14 mmol/l lower than those in the lowest fifth of plant sterol consumption (mean intake 178 mg daily); the corresponding figures in women were 0.15 and 0.13 mmol/l. After adjusting for saturated fat and fibre intakes, the results for total cholesterol and low-density lipoprotein cholesterol were similar, although the strength of the association was slightly reduced. In a free-living population, a high intake of plant sterols is inversely associated with lower concentrations of total and low-density lipoprotein serum cholesterol. The plant sterol content of foods may partly explain diet-related effects on serum cholesterol concentration.


Standardization of the Dmax method for calculating the second lactate threshold.
The purpose of this study was to test the reliability and validity of 2 standardized methods for calculating speed at the second lactate-threshold point (LT2) based on the preexisting Dmax (LTD) and modified Dmax (LTMOD) procedures. 13 trained male road runners and triathletes completed 2 incremental laboratory running tests to determine LT2, followed by separate time trials (5, 10, 15 km) on an outdoor running track. Two new methods were proposed for calculating the speed at LT2: (1) the single standardized lactate threshold (LTSDs) and (2) the paired standardized lactate threshold (LTSDp) for quantifying changes over time. The LTSDs and LTSDp methods had high relative (ICC ≥ .98) and absolute (CV ≤ 1.9%) reliability in identifying the speed at LT2. The speed at LT2 according to the LTSDs and LTSDp methods had a strong correlation and was not different to the performance speed during the 10- and 15-km time trials (≤2.3%; ρc > 0.8; P > .05). The following natural log-based formula was created to estimate the percentage of LT2 speed (using the LTSDs method) that could be sustained for events ~15-75 min: y = -7.256(ln x) + 157.64, where y = %LT2 speed, x = time-trial performance (s), and ln = natural log. The standardized methods are reliable for determining LT2. The LTSDs and LTSDp methods for calculating the speed at LT2 from a near-maximal incremental test calculated speeds similar to those exhibited in 10- and 15-km running time trials. A prediction equation for estimating the percentage of LT2 that can be sustained for events of ~15-75 min was generated.


A prospective study of dairy intake and risk of uterine leiomyomata.
Rates of uterine leiomyomata are 2-3 times higher among black women than white women. Dietary factors that differ in prevalence between these populations that could contribute to the disparity include dairy intake. During 1997-2007, the authors followed 22,120 premenopausal US Black Women's Health Study participants to assess dairy intake in relation to uterine leiomyomata risk. Because soy may be substituted for dairy, the effect of soy intake was also evaluated. Diet was estimated by using food frequency questionnaires in 1995 and 2001. Incidence rate ratios and 95% confidence intervals were estimated with Cox regression. There were 5,871 incident cases of uterine leiomyomata diagnosed by ultrasound (n = 3,964) or surgery (n = 1,907). Multivariable incidence rate ratios comparing 1, 2, 3, and > or =4 servings/day with <1 serving/day of total dairy were 0.94 (95% confidence interval (CI): 0.88, 1.00), 0.87 (95% CI: 0.78, 0.98), 0.84 (95% CI: 0.70, 1.01), and 0.70 (95% CI: 0.58, 0.86), respectively (P-trend <0.001). Incidence rate ratios comparing the highest (> or =2 servings/day) with the lowest (<1 serving/week) intake categories were 0.81 (95% CI: 0.66, 0.99) for high-fat dairy, 0.80 (95% CI: 0.70, 0.91) for low-fat dairy, and 0.78 (95% CI: 0.68, 0.89) for milk. Soy intake was unrelated to uterine leiomyomata risk. This large prospective study of black women provides the first epidemiologic evidence of reduced uterine leiomyomata risk associated with dairy consumption.


Cross-protomer interaction with the photoactive site in oligomeric proteorhodopsin complexes.
Proteorhodopsins (PRs), members of the microbial rhodopsin superfamily of seven-transmembrane-helix proteins that use retinal chromophores, comprise the largest subfamily of rhodopsins, yet very little structural information is available. PRs are ubiquitous throughout the biosphere and their genes have been sequenced in numerous species of bacteria. They have been shown to exhibit ion-pumping activity like their archaeal homolog bacteriorhodopsin (BR). Here, the first crystal structure of a proteorhodopsin, that of a blue-light-absorbing proteorhodopsin (BPR) isolated from the Mediterranean Sea at a depth of 12 m (Med12BPR), is reported. Six molecules of Med12BPR form a doughnut-shaped C6 hexameric ring, unlike BR, which forms a trimer. Furthermore, the structures of two mutants of a related BPR isolated from the Pacific Ocean near Hawaii at a depth of 75 m (HOT75BPR), which show a C5 pentameric arrangement, are reported. In all three structures the retinal polyene chain is shifted towards helix C when compared with other microbial rhodopsins, and the putative proton-release group in BPR differs significantly from those of BR and xanthorhodopsin (XR). The most striking feature of proteorhodopsin is the position of the conserved active-site histidine (His75, also found in XR), which forms a hydrogen bond to the proton acceptor from the same molecule (Asp97) and also to Trp34 of a neighboring protomer. Trp34 may function by stabilizing His75 in a conformation that favors a deprotonated Asp97 in the dark state, and suggests cooperative behavior between protomers when the protein is in an oligomeric form. Mutation-induced alterations in proton transfers in the BPR photocycle in Escherichia coli cells provide evidence for a similar cross-protomer interaction of BPR in living cells and a functional role of the inter-protomer Trp34-His75 interaction in ion transport. Finally, Wat402, a key molecule responsible for proton translocation between the Schiff base and the proton acceptor in BR, appears to be absent in PR, suggesting that the ion-transfer mechanism may differ between PR and BR.


Genomic organization of duplicated major histocompatibility complex class I regions in Atlantic salmon (Salmo salar).
We have previously identified associations between major histocompatibility complex (MHC) class I and resistance towards bacterial and viral pathogens in Atlantic salmon. To evaluate if only MHC or also closely linked genes contributed to the observed resistance we ventured into sequencing of the duplicated MHC class I regions of Atlantic salmon. Nine BACs covering more than 500 kb of the two duplicated MHC class I regions of Atlantic salmon were sequenced and the gene organizations characterized. Both regions contained the proteasome components PSMB8, PSMB9, PSMB9-like and PSMB10 in addition to the transporter for antigen processing TAP2, as well as genes for KIFC1, ZBTB22, DAXX, TAPBP, BRD2, COL11A2, RXRB and SLC39A7. The IA region contained the recently reported MHC class I Sasa-ULA locus residing approximately 50 kb upstream of the major Sasa-UBA locus. The duplicated class IB region contained an MHC class I locus resembling the rainbow trout UCA locus, but although transcribed it was a pseudogene. No other MHC class I-like genes were detected in the two duplicated regions. Two allelic BACs spanning the UBA locus had 99.2% identity over 125 kb, while the IA region showed 82.5% identity over 136 kb to the IB region. The Atlantic salmon IB region had an insert of 220 kb in comparison to the IA region containing three chitin synthase genes. We have characterized the gene organization of more than 500 kb of the two duplicated MHC class I regions in Atlantic salmon. Although Atlantic salmon and rainbow trout are closely related, the gene organization of their IB region has undergone extensive gene rearrangements. The Atlantic salmon has only one class I UCA pseudogene in the IB region while trout contains the four MHC UCA, UDA, UEA and UFA class I loci. The large differences in gene content and most likely function of the salmon and trout class IB region clearly argues that sequencing of salmon will not necessarily provide information relevant for trout and vice versa.


Site selectivity in the protonation of a phosphinito bridged Pt(I)-Pt(I) complex: a combined NMR and density-functional theory mechanistic study.
The protonation of the dinuclear phosphinito bridged complex [(PHCy2)Pt(mu-PCy2){kappa(2)P,O-mu-P(O)Cy2}Pt(PHCy2)] (Pt-Pt) (1) by Brønsted acids affords hydrido bridged Pt-Pt species the structure of which depends on the nature and on the amount of the acid used. The addition of 1 equiv of HX (X = Cl, Br, I) gives products of formal protonation of the Pt-Pt bond of formula syn-[(PHCy2)(X)Pt(mu-PCy2)(mu-H)Pt(PHCy2){kappaP-P(O)Cy2}] (Pt-Pt) (5, X = Cl; 6, X = Br; 8, X = I), containing a Pt-X bond and a dangling kappa P-P(O)Cy2 ligand. Uptake of a second equivalent of HX results in the protonation of the P(O)Cy2 ligand with formation of the complexes [(PHCy2)(X)Pt(mu-PCy2)(mu-H)Pt(PHCy2){kappaP-P(OH)Cy2}]X (Pt-Pt) (3, X = Cl; 4, X = Br; 9, X = I). Each step of protonation is reversible, thus reactions of 3, 4, with NaOH give, first, the corresponding neutral complexes 5, 6, and then the parent compound 1. While the complexes 3 and 4 are indefinitely stable, the iodine analogue 9 transforms into anti-[(PHCy2)(I)Pt(mu-PCy2)(mu-H)Pt(PHCy2)(I)] (Pt-Pt) (7) deriving from substitution of an iodo group for the P(OH)Cy2 ligand. Complexes 3 and 4 are isomorphous crystallizing in the triclinic space group P1 and show an intramolecular hydrogen bond and an interaction between the halide counteranion and the POH hydrogen. The occurrence of such an interaction also in solution was ascertained for 3 by (35)Cl NMR. Multinuclear NMR spectroscopy (including (31)P-(1)H HOESY) and density-functional theory calculations indicate that the mechanism of the reaction starts with a prior protonation of the oxygen with formation of an intermediate (12) endowed with a six membered Pt(1)-X...H-O-P-Pt(2) ring that evolves into thermodynamically stable products featuring the hydride ligand bridging the Pt atoms. Energy profiles calculated for the various steps of the reaction between 1 and HCl showed very low barriers for the proton transfer and the subsequent rearrangement to 12, while a barrier of 29 kcal mol(-1) was found for the transformation of 12 into 5.


Skeletrophin, a novel RING molecule controlled by the chromatin remodeling complex, is downregulated in malignant melanoma.
Recent experiments have revealed that aberrant functionality of the chromatin remodeling complex is related to tumorigenicity in various malignant tumors. Skeletrophin is an actin-binding cytoskeleton-related molecule, which is induced by the overexpression of truncated human SWI1 (SMARCF1). Human SWI1 is a sub-unit of the chromatin remodeling complex and binds chromatin through its ARID (AT-rich interactive domain). Truncated SWI1 lacks one of the two glucocorticoid-receptor binding domains and inhibits the intact human SWI1 in a dominant negative manner. Skeletrophin, was therefore identified as a candidate molecule for the indication of change to a malignant phenotype due to the aberrant function of the chromatin remodeling complex. Surprisingly, the skeletrophin gene is located in 1p36.32, where the putative tumor suppressor gene of cutaneous malignant melanoma has long been postulated to be on. Cutaneous malignant melanoma is a highly aggressive tumor. To overcome the clinical problem of malignant melanoma and highly invasive and metastatic activity, it is important to unravel the molecular mechanism responsible for melanoma progression. Recent studies including those from our laboratories have elucidated that skeletrophin is a novel RING-HC type ubiquitin ligase and that the ubiquitin ligase pathway mediated by skeletrophin acts to oppose melanoma cell invasion. Here, we summarize the characterization of skeletrophin, with emphasis on its biological activity, the disruption of which is linked with melanoma progression.


Temperature controlled high-throughput magnetic tweezers show striking difference in activation energies of replicating viral RNA-dependent RNA polymerases.
RNA virus survival depends on efficient viral genome replication, which is performed by the viral RNA dependent RNA polymerase (RdRp). The recent development of high throughput magnetic tweezers has enabled the simultaneous observation of dozens of viral RdRp elongation traces on kilobases long templates, and this has shown that RdRp nucleotide addition kinetics is stochastically interrupted by rare pauses of 1-1000 s duration, of which the short-lived ones (1-10 s) are the temporal signature of a low fidelity catalytic pathway. We present a simple and precise temperature controlled system for magnetic tweezers to characterize the replication kinetics temperature dependence between 25°C and 45°C of RdRps from three RNA viruses, i.e. the double-stranded RNA bacteriophage Φ6, and the positive-sense single-stranded RNA poliovirus (PV) and human rhinovirus C (HRV-C). We found that Φ6 RdRp is largely temperature insensitive, while PV and HRV-C RdRps replication kinetics are activated by temperature. Furthermore, the activation energies we measured for PV RdRp catalytic state corroborate previous estimations from ensemble pre-steady state kinetic studies, further confirming the catalytic origin of the short pauses and their link to temperature independent RdRp fidelity. This work will enable future temperature controlled study of biomolecular complex at the single molecule level.


[The layperson in emergency situation].
The layperson's qualification to provide early care in emergency situations and basic life support (BLS) is fundamental to save lives and prevent sequels. The objective was to identify the level of knowledge of lay people about approaching an emergency victim. Structured interviews in non-technical language were used with a 385-subject sample, average age 35.4 (+/- 14.55) years, with more than 50% having a high school or university education. Over 55% of these observed situations with loss of consciousness, but only 31% called for specialized help. 34% underwent a first-aid course, but only 13% feel prepared. The most often cited place of learning was the driving school (DS), 35.9%. Other training places were higher when compared to the DS (p = 0.048). Almost 17% do not know how to recognize the presence of vital signs. Almost 31% do not know the telephone number of the emergency service. Laypersons have incomplete or incorrect knowledge about care for unconscious victims.


Language-independent automatic evaluation of intelligibility of chronically hoarse persons.
Automatic intelligibility assessment using automatic speech recognition is usually language specific. In this study, a language-independent approach is proposed. It uses models that are trained with Flemish speech, and it is applied to assess chronically hoarse German speakers. The research questions are here: is it possible to construct suitable acoustic features that generalize to other languages and a speech disorder, and is the generated model for intelligibility also suitable for specific subtypes of that disorder, i.e. functional and organic dysphonia? 73 German-speaking persons with chronic hoarseness read the text 'Der Nordwind und die Sonne'. Perceptual intelligibility scores were used as ground truth during the training of an automatic model that converts speaker level acoustic measurements into intelligibility scores. Cross-validation is used to assess model performance. The interrater agreement for all patients (n = 73) and for the functional and organic dysphonia subgroups (n = 45 and n = 24) are r = 0.82, r = 0.83 and r = 0.75, respectively. The automatic assessment based on phonologically based acoustic models revealed correlations between perceptual and automatic intelligibility ratings of r = 0.79 (all patients), r = 0.78 (functional dysphonia) and r = 0.80 (organic dysphonia). The automatic, objective measurement of intelligibility is a valuable instrument in an evidence-based clinical practice.


The cost of ethics legislation: a look at the Patient Self-Determination Act.
The Patient Self-Determination Act (PSDA) requires hospitals to ask patients upon admission whether they have an advance directive. Although the PSDA has received extensive criticism, little attention has been paid to the cost of the law, either during its legislative course or following its implementation. Nonetheless, several tangible and intangible costs are associated with the PSDA. Such costs may be incurred by different parties. This paper examines the costs and benefits of the PSDA and illustrates the extent of some of its tangible costs. The incremental start-up cost for one institution's response to the PSDA is estimated to be $49,304 ($1.31 per admission) and the total implementation cost of the program to be $114,528. In addition, the national incremental start-up cost for hospitals to implement the PSDA is estimated to be between $43,625,114 and $101,569,922. Finally, the potential implications of the PSDA for future governmental health care ethics regulation are discussed.


Waiting list eradication in secondary care psychology: Addressing a National Health Service blind spot.
Waiting times for secondary care psychological therapy remain a 'blind spot' in serious mental illness (SMI) provision, and their reduction is a priority within the National Health Service (NHS) Five Year Forward View. The paper describes the eradication of waiting times within a community-based NHS service and the effectiveness of strategies whilst examining help-seeking behaviour, compliance and therapeutic need. Analyses are reported for treatment compliance and therapeutic outcomes for 208 waiting-list cohort individuals seen by the SMI psychology service over an 18-month period between October 2014 and March 2016. No significant clinical or demographic differentiation between individuals who successfully completed therapy compared to those who disengaged was observed. Despite an average 2.20-year waiting time, this alone did not significantly impact engagement with psychological treatment and all psychological therapies provided led to a significant clinical improvement and no individuals who completed therapy required re-referral at 12-month follow-up. If imposed appropriately over a suitable time frame evidence-based practice coupled with effective operationalization can result in efficient needs-led psychological provision within SMI and secondary care. Potentially debilitating waiting times for service users and other referring professionals can be avoided, whilst psychology provision retains a flexible, formulation-based and person-centred approach.


Considerations for developing and implementing an online community-based exercise intervention with adults living with HIV: a qualitative study.
To describe the need for, and utility of, online community-based exercise (CBE) interventions with adults living with HIV and identify factors to consider in developing and implementing an online CBE intervention with adults living with HIV. Qualitative descriptive study using web-based semistructured interviews. We recruited adults representing at least one of five stakeholder groups with experience in CBE and/or HIV: (1) adults living with HIV, (2) rehabilitation professionals, (3) fitness personnel, (4) educators with eLearning experience and (5) representatives from HIV community-based organisations (CBOs). We asked participants to describe their experiences with online CBE, need and utility for online CBE and factors in developing and implementing online CBE interventions. We analysed data using a group-based thematic analytical approach. Among the 11 participants, most had experience working with adults living with HIV (73%) or with telehealth/rehabilitation/coaching in HIV or other chronic conditions (91%). Participants (eight women; two men; one non-binary; median age: 49 years), identified the need and utility for online CBE interventions to increase accessibility and continuity of care with adults living with HIV. Six factors to consider in developing and implementing online CBE included: (1) person-specific considerations (episodic nature of HIV, stigma, HIV disclosure), (2) accessibility of programme (physical space to exercise, reliable internet, access to devices, digital literacy), (3) programme delivery and technology (live vs prerecorded online classes, multiple online platforms for delivery, physical activity tracking, troubleshooting technology), (4) attributes of programme personnel (working with CBOs, relatable instructors, diverse staff), (5) programme content and design (tailored exercise classes, educational sessions) and (6) building community (shared experiences, peer support, social opportunities). There is a need and utility for online CBE in the context of HIV. Considerations for development and implementation span individual, structural and technical, and community dimensions. Results can inform the future development and implementation of online CBE with adults living with HIV and other chronic episodic conditions.


SMG-2 is a phosphorylated protein required for mRNA surveillance in Caenorhabditis elegans and related to Upf1p of yeast.
mRNAs that contain premature stop codons are selectively degraded in all eukaryotes tested, a phenomenon termed "nonsense-mediated mRNA decay" (NMD) or "mRNA surveillance." NMD may function to eliminate aberrant mRNAs so that they are not translated, because such mRNAs might encode deleterious polypeptide fragments. In both yeasts and nematodes, NMD is a nonessential system. Mutations affecting three yeast UPF genes or seven nematode smg genes eliminate NMD. We report here the molecular analysis of smg-2 of Caenorhabditis elegans. smg-2 is homologous to UPF1 of yeast and to RENT1 (also called HUPF1), a human gene likely involved in NMD. The striking conservation of SMG-2, Upf1p, and RENT1/HUPF1 in both sequence and function suggests that NMD is an ancient system, predating the divergence of most eukaryotes. Despite similarities in the sequences of SMG-2 and Upf1p, expression of Upf1p in C. elegans does not rescue smg-2 mutants. We have prepared anti-SMG-2 polyclonal antibodies and identified SMG-2 on Western blots. SMG-2 is phosphorylated, and mutations of the six other smg genes influence the state of SMG-2 phosphorylation. In smg-1, smg-3, and smg-4 mutants, phosphorylation of SMG-2 was not detected. In smg-5, smg-6, and smg-7 mutants, a phosphorylated isoform of SMG-2 accumulated to abnormally high levels. In smg-2(r866) and smg-2(r895) mutants, which harbor single amino acid substitutions of the SMG-2 nucleotide binding site, phosphorylated SMG-2 accumulated to abnormally high levels, similar to those observed in smg-5, smg-6, and smg-7 mutants. We discuss these results with regard to the in vivo functions of SMG-2 and NMD.


[Giant condyloma (Buschke-Löwenstein) of the vulva].
Diagnosis, therapy and biological behaviour of the Buschke-Löwenstein tumour of the vulva are described. Surgical treatment is accompanied by an antiviral therapy with interferon. Viral genesis of HPV 6 and/or HPV 11 is probably the cause of this intermediate tumour a type between condyloma and invasive squamous cell carcinoma.


Hemoglobinopathy screening in pregnancy: comparison of two protocols.
This study was designed to determine the ability of a hemoglobinopathy screening protocol involving sickle solubility testing and red blood cell (RBC) indices to identify at-risk pregnancies. Retrospective chart review of all patients registering for prenatal care at the New York Hospital/Cornell Medical Center prenatal clinic in 1996 was the study design. All patients had RBC indices as well as hemoglobin electrophoresis. RBC indices of those with normal and abnormal electrophoreses were compared. Comparison of protocols involving universal hemoglobin electrophoresis and selective use of hemoglobin electrophoresis were compared. Student's t-test was used for statistical analysis. There were 36 carriers of hemoglobinopathy traits in 631 patients screened (5.7%). Four (three with hemoglobin C trait and one with hemoglobin D trait) had normal RBC indices and presumably would have had negative sickle solubility testing. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of a protocol with selective use of hemoglobin electrophoresis would have been 88.9, 79.4, 20.8, and 99.2%, respectively. Cost analysis reveals a difference of $11,384 or $18/patient less in a protocol with selective use of hemoglobin electrophoresis. Although a protocol involving solubility testing with RBC indices will not identify every carrier of a hemoglobinopathy trait, it may be appropriate in some populations.


Fred C. Cuny Memorial Continuing Education Series. Lesson 10: group dynamics in disasters: managing work groups.
Groups are assigned or formed to perform tasks that one person cannot accomplish alone. This lesson describes the classification of work groups, group unity, leadership, motivation, recognition, conflict resolution, and remediation associated with managing groups and their activities. Advantages associated with group process include 1) the generation of better ideas, 2) ability to assume greater risks; make fewer errors; 3) the capacity for greater knowledge and 4) information, and for some problems, production of better decisions. Groups may be formal or informal. Formal groups may be organic, task-directed, or committees. Informal groups arise when it becomes obvious that a group will work better or may be formed by a discipline within the organization or through friendships. The size of the group its status within the organization, the goals established, and the dependence of the members on the group all may affect the cohesiveness of the group. Leadership of the group must keep the group focused on the objectives and enhancement of the efficiency of its operation and the quality of the decisions made by the group. Motivation of a group often is more difficult than is that for individuals and generally positive inducements work better than do negative measures such as coercion and reprimands. Roles are often informal and conferred by the group collectively. Often norms are established within groups that help the group deal with conflict. Inadequate performance within a formal group may require changes in the leadership, removal of a member or clique within the group, reduction of group size, and/or dissolution of the group. Understanding the dynamics of groups is an essential skill required of good managers.


Adalimumab in the treatment of immune-mediated diseases.
Tumour necrosis factor (TNF) plays an important role in the pathogenesis of immune-mediated inflammatory diseases (IMIDs). TNF inhibition results in down-regulation of abnormal and progressive inflammatory processes, resulting in rapid and sustained clinical remission, improved quality of life and prevention of target organ damage. Adalimumab is the first fully human monoclonal antibody directed against TNF. In this article, we review the role and cost effectiveness of adalimumab in the treatment of IMIDs in adults and children. The efficacy and tolerability of adalimumab has been demonstrated in patients with a wide range of inflammatory conditions, leading to regulatory approval in rheumatoid arthritis (RA), psoriatic arthritis (PsA), plaque psoriasis, inflammatory bowel diseases (Crohn's disease, ulcerative colitis, paediatric Crohn's disease, and intestinal Behçet's disease), ankylosing spondylitis (AS), axial spondyloarthritis (SpA) and juvenile idiopathic arthritis. The major tolerability issues with adalimumab are class effects, such as injection site reactions and increased risk of infection and lymphoma. As with all anti-TNF agents, adalimumab is immunogenic, although less than infliximab, and some patients receiving long-term adalimumab will develop anti-drug antibodies, causing a loss of response. Comparisons of its clinical utility and cost effectiveness have shown it to be a valid treatment choice in a wide range of patients. Recent data from Italian economic studies show the cost effectiveness of adalimumab to be below the threshold value for health care interventions for most indications. In addition, analysis of indirect costs shows that adalimumab significantly reduces social costs associated with RA, PsA, AS, Crohn's disease and psoriasis. The fact that adalimumab has the widest range of approved indications, many often presenting together in the same patient due to the common pathogenesis, may further improve the utility of adalimumab. Current clinical evidence shows adalimumab to be a valuable resource in the management of IMIDs. Further research, designed to identify patients who may benefit most from this drug, will better highlight the role and cost-effectiveness of this versatile TNF inhibitor.


Safe Injection Practices in Primary Health Care Settings of Naxalbari Block, Darjeeling District, West Bengal.
Unsafe injection can transmit many diseases to patients, injection providers and healthy people of community. To find out critical steps whether executed according to recommended best practice methods, availability of equipments in health facilities for safe injection practices and some important steps of waste disposal methods. This facility-based cross-sectional observational study was conducted among 30 Auxiliary nurse midwives (ANM) & 27 nursing staffs (NS) to assess certain aspects of their practice while administrating injection and disposal of the disposables. Health facilities were also observed to asses necessary equipments of safe injection and waste disposal methods. Among the health workers 93.3% ANM and 100% NS took sterile syringe from sterile unopened packet, all of the study subjects washed hand before giving injection, 13.3% of ANMs and 8% of NS are fully vaccinated against Hep B, 53.3% of ANM and all NS are practices non recapping. Only 13.33% sub centres along with PHC & BPHC had at least one puncture resistant leak proof container, 86.7% sub centres, PHC are free from loose needles. Transport for off side treatment is the method of waste disposal in case of 73.3% cases sub centres, PHC & BPHC. There is need to educate, train and motivate service providers in proper methods of giving injection along with improve the adequacy of supply of required equipments.


Whole Genome Sequencing Refines Knowledge on the Population Structure of Mycobacterium bovis from a Multi-Host Tuberculosis System.
Classical molecular analyses of Mycobacterium bovis based on spoligotyping and Variable Number Tandem Repeat (MIRU-VNTR) brought the first insights into the epidemiology of animal tuberculosis (TB) in Portugal, showing high genotypic diversity of circulating strains that mostly cluster within the European 2 clonal complex. Previous surveillance provided valuable information on the prevalence and spatial occurrence of TB and highlighted prevalent genotypes in areas where livestock and wild ungulates are sympatric. However, links at the wildlife-livestock interfaces were established mainly via classical genotype associations. Here, we apply whole genome sequencing (WGS) to cattle, red deer and wild boar isolates to reconstruct the M. bovis population structure in a multi-host, multi-region disease system and to explore links at a fine genomic scale between M. bovis from wildlife hosts and cattle. Whole genome sequences of 44 representative M. bovis isolates, obtained between 2003 and 2015 from three TB hotspots, were compared through single nucleotide polymorphism (SNP) variant calling analyses. Consistent with previous results combining classical genotyping with Bayesian population admixture modelling, SNP-based phylogenies support the branching of this M. bovis population into five genetic clades, three with apparent geographic specificities, as well as the establishment of an SNP catalogue specific to each clade, which may be explored in the future as phylogenetic markers. The core genome alignment of SNPs was integrated within a spatiotemporal metadata framework to further structure this M. bovis population by host species and TB hotspots, providing a baseline for network analyses in different epidemiological and disease control contexts. WGS of M. bovis isolates from Portugal is reported for the first time in this pilot study, refining the spatiotemporal context of TB at the wildlife-livestock interface and providing further support to the key role of red deer and wild boar on disease maintenance. The SNP diversity observed within this dataset supports the natural circulation of M. bovis for a long time period, as well as multiple introduction events of the pathogen in this Iberian multi-host system.


Complement regulates inhalation tolerance at the dendritic cell/T cell interface.
Pulmonary exposure to innocuous aeroallergens is a common event leading to inhalation tolerance. Distinct subsets of pulmonary dendritic cells (DC) and regulatory T cells (T(Reg)) play critical roles in mediating and maintaining such tolerance. In asthmatics, the same aeroallergens drive a maladaptive, Th2-biased immune response resulting in airway inflammation and airway hyper-reactivity. The mechanisms underlying the breakdown of inhalation tolerance, leading to the Th2-driven inflammation in rising numbers of asthmatic patients from industrialized countries remain elusive. The recent resurgence of interest in the role of the innate immune mediators in regulating adaptive immune response has sparked studies aimed at identifying the role of complement in allergic asthma. In this context, an unexpected role for the anaphylatoxin C5a receptor in allergic sensitization has been found. In models of experimental allergic asthma, ablation of C5aR signaling during initial allergen exposure either induced or enhanced Th2 sensitization. Mechanistically, C5aR signaling directly affected the function of distinct pulmonary DC subsets that induce or control allergen-induced adaptive immune responses. Signaling pathways downstream of C5 may also impact the function of T(Reg), as T(Reg) from C5 sufficient, but not from C5 deficient mice, suppress DC activation and subsequent development of Th2-driven inflammation. The emerging paradigm is that constitutive local generation of C5a and C5aR signaling in airway DCs controls inhalation tolerance directly as well as indirectly through sensitization of airway DCs for T(Reg)-mediated immunosuppression.


Klebsiella pneumoniae necrotizing fasciitis of the leg in an elderly French woman.
Klebsiella pneumoniae necrotizing fasciitis is a rare infection in regions outside of Asia. Here, we present a case of necrotizing fasciitis of the leg caused by K. pneumoniae in a 92-year-old French woman hospitalized in a geriatric rehabilitation unit. The patient initially presented with dermohypodermitis of the leg that developed from a dirty wound following a fall. A few hours later, this painful injury extended to the entire lower limb, with purplish discoloration of the skin, bullae, and necrosis. Septic shock rapidly appeared and the patient died 9 hours after the onset of symptoms. The patient was Caucasian, with no history of travel to Asia or any underlying disease. Computed tomography revealed no infectious metastatic loci. Blood cultures showed growth of capsular serotype K2 K. pneumoniae strains with virulence factors RmpA, yersiniabactin and aerobactin. This rare and fatal case of necrotizing fasciitis caused by a virulent strain of K. pneumoniae occurred in a hospitalized elderly woman without risk factors. Clinicians and geriatricians in particular should be aware of this important albeit unusual differential diagnosis.


[Behavior of the primary care physicians in the treatment of community infections].
This study aimed to evaluate the behavior of the physicians who attend to patients with community infections who are responsible for most of the antibiotic prescriptions made in the community setting. Furthermore, the evolution over the last 10 years in regards to the treatment habits of the most frequent infections in Primary Health Care (PHC) was studied and prescription behaviors were analyzed in relationship to the acquaintance and attitude of the participating physicians. A data collection sheet from 1,411 prescriptions made by 855 physicians from different specialties was used to conduct the study. This represents a statistically significant sample nationwide both from the point of view of the physicians involved in the treatment of infectious diseases in PHC as well as regarding to the prescriptions they have written. The study reveals the amplitude of respiratory and pararespiratory disease with oral antimicrobial agents, particularly beta-lactamics, and the significant advance in the last 10 years in the use of amoxicillin and clavulanic acid as treatment of choice. Treatment duration was somewhat longer while the number of containers prescribed was less than in the previous study. This would be related to the use of more spaced doses of the antimicrobial agents. Furthermore, the study reveals some discrepancies between attitudes and events of the physicians in the approach to certain diagnoses. Key words: Behavior. Primary care physicians. Prescription. Antimicrobials agents.


Panic plaques: panic disorder & coronary artery disease in patients with chest pain.
The purpose of this systematic review was to identify characteristics of the chest pain associated with the presence of panic disorder, to determine the strength of the association between panic disorder and coronary artery disease (CAD), and to determine the association between panic disorder and known cardiovascular risk factors. Potential studies were identified via computerized search using MEDLINE and PSYCINFO databases, and review of bibliographies. MeSH headings used included "panic disorder" with "chest pain," "panic disorder" with "coronary disease or cardiovascular disorders or heart disorders," and "panic disorder" with "cholesterol or essential hypertension or tobacco smoking." Studies had to base their diagnosis of panic disorder on criteria from the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, and objective criteria of CAD and risk factors had to be used. Only case-control and cohort studies were included. The relative risk of panic disorder in patients with nonanginal chest pain is 2.03 [confidence interval (CI), 1.41 to 2.92]. Concerning the relationship between panic disorder and CAD, studies conducted in emergency departments found a relative risk of 1.25 (CI, 0.87 to 1.80). However, there is an inverse relationship between the prevalence of CAD in the study and the prevalence of panic disorder among the patients with CAD (r = -.469, P =.086). Panic disorder has also been linked to cardiac risk factors. Panic disorder and CAD are correlated in noncardiology settings, and recurrent panic attacks may actually cause CAD. Recognition of either condition should lead the family physician to consider the other, resulting in increased vigilance and possible screening.


Growth-regulatory human galectin-1: crystallographic characterisation of the structural changes induced by single-site mutations and their impact on the thermodynamics of ligand binding.
Human galectin-1 is a potent multifunctional effector that participates in specific protein-carbohydrate and protein-protein (lipid) interactions. By determining its X-ray structure, we provide the basis to define the structure of its ligand-binding pocket and to perform rational drug design. We have also analysed whether single-site mutations introduced at some distance from the carbohydrate recognition domain can affect the lectin fold and influence sugar binding. Both the substitutions introduced in the C2S and R111H mutants altered the presentation of the loop, harbouring Asp123 in the common "jelly-roll" fold. The orientation of the side-chain was inverted 180 degrees and the positions of two key residues in the sugar-binding site of the R111H mutant were notably shifted, i.e. His52 and Trp68. Titration calorimetry was used to define the decrease in ligand affinity in both mutants and a significant increase in the entropic penalty was found to outweigh a slight enhancement of the enthalpic contribution. The position of the SH-groups in the galectin appeared to considerably restrict the potential to form intramolecular disulphide bridges and was assumed to be the reason for the unstable lectin activity in the absence of reducing agent. However, this offers no obvious explanation for the improved stability of the C2S mutant under oxidative conditions. The noted long-range effects in single-site mutants are relevant for the functional divergence of closely related galectins and in more general terms, the functionality definition of distinct amino acids.


Lewis adducts of the side-on end-on dinitrogen-bridged complex [{(NPN)Ta}2(mu-H)2(mu-eta 1:eta 2-N2)] with AlMe3, GaMe3, and B(C6F5)3: synthesis, structure, and spectroscopic properties.
Reaction of the side-on end-on dinitrogen complex [{(NPN)Ta}(2)(mu-H)(2)(mu-eta(1):eta(2)-N(2))] (1; in which NPN=(PhNSiMe(2)CH(2))(2)PPh), with the Lewis acids XR(3) results in the adducts [{(NPN)Ta}(2)(mu-H)(2)(mu-eta(1):eta(2)-NNXR(3))], XR(3)=GaMe(3) (2), AlMe(3) (3), and B(C(6)F(5))(3) (4). The solid-state molecular structures of 2, 3, and 4 demonstrate that the N-N bond length increases relative to those found in 1 by 0.036, 0.043, and 0.073 A, respectively. In solution complexes 2-4 are fluxional as evidenced by variable-temperature (1)H NMR spectroscopy. The (15)N{(1)H} NMR spectra of 2-4 are reported; furthermore, their vibrational properties and electronic structures are evaluated. The vibrational structures are found to be closely related to that of the parent complex 1. Detailed spectroscopic analysis on 2-4 leads to the identification of the theoretically expected six normal modes of the Ta(2)N(2) core. On the basis of experimental frequencies and the QCB-NCA procedure, the force constants are determined. Importantly, the N-N force constant decreases from 2.430 mdyn A(-1) in 1 to 1.876 (2), 1.729 (3), and 1.515 mdyn A(-1) (4), in line with the sequence of N-N bond lengths determined crystallographically. DFT calculations on a generic model of the Lewis acid adducts 2-4 reveal that the major donor interaction between the terminal nitrogen atom and the Lewis acid is mediated by a sigma/pi hybrid molecular orbital of N(2), corresponding to a sigma bond. Charge analysis performed for the adducts indicates that the negative charge on the terminal nitrogen atom of the dinitrogen ligand increases with respect to 1. The lengthening of the N-N bond observed for the Lewis adducts is therefore explained by the fact that charge donation from the complex fragment into the pi* orbitals of dinitrogen is increased, while electron density from the N-N bonding orbitals p(sigma) and pi(h) is withdrawn due to the sigma interaction with the Lewis acid.


Machine Learning Interpretation of Extended Human Papillomavirus Genotyping by Onclarity in an Asian Cervical Cancer Screening Population.
This study aimed (i) to compare the performance of the BD Onclarity human papillomavirus (HPV) assay with the Cobas HPV test in identifying cervical intraepithelial neoplasia 2/3 or above (CIN2/3+) in an Asian screening population and (ii) to explore improving the cervical cancer detection specificity of Onclarity by machine learning. We tested 605 stratified random archived samples of cervical liquid-based cytology samples with both assays. All samples had biopsy diagnosis or repeated negative cytology follow-up. Association rule mining (ARM) was employed to discover coinfection likely to give rise to CIN2/3+. Outcome classifiers interpreting the extended genotyping results of Onclarity were built with different underlying models. The sensitivities (Onclarity, 96.32%; Cobas, 95.71%) and specificities (Onclarity, 46.38%; Cobas, 45.25%) of the high-risk HPV (hrHPV) components of the two tests were not significantly different. When HPV16 and HPV18 were used to further interpret hrHPV-positive cases, Onclarity displayed significantly higher specificity (Onclarity, 87.10%; Cobas, 80.77%). Both hrHPV tests achieved the same sensitivities (Onclarity, 90.91%; Cobas, 90.91%) and similar specificities (Onclarity, 48.46%; Cobas, 51.98%) when used for triaging atypical squamous cells of undetermined significance. Positivity in both HPV16 and HPV33/58 of the Onclarity channels entails the highest probability of developing CIN2/3+. Incorporating other hrHPVs into the outcome classifiers improved the specificity of identifying CIN2/3 to up to 94.32%. The extended genotyping of Onclarity therefore can help to highlight patients having the highest risk of developing CIN2/3+, with the potential to reduce unnecessary colposcopy and negative psychosocial impact on women receiving the reports.


Delayed complications of treated reduced recurrent anterior dislocations of the humeral head in young adults.
Patients with recurrent anterior dislocations of the humeral head are advised to have the disability corrected by open surgical intervention. Painful and restricted motion after such a procedure is not always due to recurrence of the dislocation; it may be a consequence of changes in the articular surfaces and soft tissues of the joint-i.e., osteoarthritis. We observed that osteoarthritis was present in instances of recurrent anterior dislocations in both operative and nonoperative cases, but to a lesser degree in some of the operative cases.


Meta-analysis of survival prediction with Palliative Performance Scale.
This paper aims to reconcile the use of Palliative Performance Scale (PPSv2) for survival prediction in palliative care through an international collaborative study by five research groups. The study involves an individual patient data meta-analysis on 1,808 patients from four original datasets to reanalyze their survival patterns by age, gender, cancer status, and initial PPS score. Our findings reveal a strong association between PPS and survival across the four datasets. The Kaplan-Meier survival curves show each PPS level as distinct, with a strong ordering effect in which higher PPS levels are associated with increased length of survival. Using a stratified Cox proportional hazard model to adjust for study differences, we found females lived significantly longer than males, with a further decrease in hazard for females not diagnosed with cancer. Further work is needed to refine the reporting of survival times/probabilities and to improve prediction accuracy with the inclusion of other variables in the models.


Diethylnitrosamine induced enzyme altered areas in the rat liver: effect of partial hepatectomy.
The effect of partial hepatectomy on the growth of enzyme-altered areas, induced in male albino Wistar rats by giving them diethylnitrosamine (DEN) in their drinking water, was studied. This was performed morphometrically by means of light microscopy and enzyme histochemistry. The results showed that the enzyme-altered areas increased significantly in size for two weeks following the partial hepatectomy but not thereafter. This finding suggested that DEN induced enzyme-altered areas can respond to a proliferative stimulus, but the response appeared transient.


Leptomeningeal metastasis after surgical resection of brain metastases.
To determine the incidence and risk factors for leptomeningeal metastasis after surgery for brain metastasis of solid tumors. Review of the records of all patients operated on for brain metastasis between January 1990 and August 1995. In this period 28 patients underwent surgery for brain metastasis, of whom 27 were available for evaluation in this study. Median survival after craniotomy was 11 months. Nine patients (33%) developed leptomeningeal metastasis 2-13 months after surgery, which included six of the nine patients operated on for posterior fossa metastasis (p=0.05). In five patients, leptomeningeal metastasis was the only site of recurrence. Three patients developed the leptomeningeal metastasis as bulky tumour along the spinal cord, which is a rare presentation. No other risk factors for the development of leptomeningeal metastasis other than surgery for posterior fossa metastasis were identified. There is an increased risk of leptomeningeal metastasis after surgery for posterior fossa metastasis. Future trials should consider the value of an active approach to this complication in these patients.


A mathematical formulation for the cell-cycle model in somitogenesis: analysis, parameter constraints and numerical solutions.
In this work we present an analysis, supported by numerical simulations, of the formulation of the cell-cycle model for somitogenesis proposed in Collier et al. (J. Theor Biol. 207 (2000), 305-316). The analysis indicates that by introducing appropriate parameter constraints on model parameters the cell-cycle mechanism can indeed give rise to the periodic pattern of somites observed in normal embryos. The analysis also provides a greater understanding of the signalling process controlling somite formation and allows us to understand which parameters influence somite length.


Knowledge of Clinical Trial Availability and Reasons for Nonparticipation Among Adolescent and Young Adult Cancer Patients: A Population-based Study.
Adolescent and young adult (AYA) cancer patients are underrepresented in clinical trials, but the reasons for this phenomenon are unknown. Questionnaire and medical record data from 515 AYA cancer patients (21 acute lymphocytic leukemia [ALL], 201 germ cell tumor, 141 Hodgkin lymphoma, 128 non-Hodgkin lymphoma, 24 sarcoma) from a population-based study were analyzed. We used multivariable models to determine characteristics associated with patient knowledge of the availability of clinical trials for their cancer. Reasons for not participating in a trial were tabulated. In total, 63% of patients reported not knowing whether a relevant clinical trial was available, 20% reported knowing that a clinical trial was not available, and 17% reported that a trial was available. Among patients reporting an available trial, 67% were recommended for enrollment. Knowing about the availability of clinical trials was associated with having ALL (odds ratio=2.9, 95% confidence interval=1.1, 7.8). Reporting that a clinical trial was available was positively associated with having ALL, Hodgkin lymphoma, non-Hodgkin lymphoma and sarcoma (relative to germ cell tumor) and working full-time or in school full-time (odds ratio=2.6, 95% confidence interval=1.0, 6.7). Concerns about involvement in research (57%) and problems accessing trials (21%) were the primary reasons cited for not enrolling among patients who knew that a trial was available. Improvement in AYA cancer patient clinical trial enrollment will require enhancing knowledge about trial availability and addressing this population's concerns about participating in medical research.


Expression of the cholinergic gene locus in pulmonary arterial endothelial cells.
In the pulmonary vasculature of man, pig and guinea-pig, acetylcholine (ACh) exerts a relaxant effect by interacting with muscarinic receptors located on endothelial cells. The present experiments were designed to detect the endogenous source of ACh in the pulmonary vasculature. For this purpose, we investigated whether pulmonary artery endothelial cells contain elements of the "cholinergic gene locus", the ACh synthesising enzyme choline acetyltransferase (ChAT) and the vesicular ACh transporter (VAChT). ChAT mRNA was detected by means of reverse transcription polymerase chain reaction (RT-PCR) in endothelial cells of porcine pulmonary arteries freshly isolated and after 7 days in culture. ChAT protein was demonstrated in endothelial cells in vitro and in situ. ChAT immunoreactivity was present in endothelial cells freshly isolated and after 2, 4, 7, and 9 days in culture. Tissue sections from extra- and intraparenchymal pulmonary arteries of man, pig and guinea-pig expressed a mosaic pattern of ChAT-positive and -negative endothelial cells. VAChT mRNA was detected by RT-PCR in rat pulmonary artery and in endothelial cells isolated from human and porcine pulmonary trunk. The detection of VAChT and ChAT mRNA and the demonstration of ChAT protein in vitro and in situ suggest that the endothelium is an endogenous source of ACh in the pulmonary vasculature.


[Agmatine--an endogenous ligand of imidazoline receptors].
The existence of imidazoline receptors was suggested based on the results of investigation of the central hypotensive action of the alpha 2-adrenoreceptor agonist clonidine containing imidazoline group in the molecule structure. Experiments on the rostral venterolateral medulla oblongata (RVLM) neurons in rats showed that not less that 36% of specific binding sites in this zone differ from adrenergic and recognize imidazoline derivatives. Agmatine has proved to be an endogenous ligand of such imidazoline receptors.


Maternal prenatal cortisol and the interaction of income and pre-pregnancy body mass index are independently associated with newborn cortisol.
While extensive research has supported the developmental programming hypothesis regarding contributions of prenatal psychosocial or nutritional adversity to offspring stress physiology, fewer studies consider both exposures together with maternal stress physiology. This study examined newborn cortisol output during a stressor as a function of maternal pre-pregnancy health status and nutritional history (pre-pregnancy body mass index [PPBMI]), economic resources (household income), and maternal cortisol awakening response (mCAR) in late pregnancy. Participants were 102 mother-infant pairs from an economically and racial/ethnically diverse sample. Offspring salivary cortisol response to a neurobehavioral exam was assessed at 1 month. Income and maternal PPBMI were positively associated with mCAR in late pregnancy. mCAR was positively related to 1-month newborn cortisol response. The interaction of income and PPBMI was positively associated with newborn cortisol output during an exam at 1-month. Mothers with the highest PPBMI and lowest income had offspring with higher cortisol responses than offspring of mothers with higher income and lower PPBMI. There was no evidence of indirect mediation effects of predictors (PPBMI, income, and interaction) on infant cortisol via mCAR. The differential effects of the interaction of PPBMI and income suggest that these exposures influence infant cortisol output in the context of one another, independent of maternal pregnancy cortisol.


Multiproxy bioarchaeological data reveals interplay between growth, diet and population dynamics across the transition to farming in the central Mediterranean.
The transition to farming brought on a series of important changes in human society, lifestyle, diet and health. The human bioarchaeology of the agricultural transition has received much attention, however, relatively few studies have directly tested the interrelationship between individual lifestyle factors and their implications for understanding life history changes among the first farmers. We investigate the interplay between skeletal growth, diet, physical activity and population size across 30,000 years in the central Mediterranean through a 'big data' cross-analysis of osteological data related to stature (n = 361), body mass (n = 334) and long bone biomechanics (n = 481), carbon (δ13C) and nitrogen (δ15N) stable isotopes (n = 1986 human, n = 475 animal) and radiocarbon dates (n = 5263). We present the observed trends on a continuous timescale in order to avoid grouping our data into assigned 'time periods', thus achieving greater resolution and chronological control over our analysis. The results identify important changes in human life history strategies associated with the first farmers, but also highlight the long-term nature of these trends in the millennia either side of the agricultural transition. The integration of these different data is an important step towards disentangling the complex relationship between demography, diet and health, and reconstruct life history changes within a southern European context. We believe the methodological approach adopted here has broader global implications for bioarchaeological studies of human adaptation more generally.


Adherence to the Sydney System guidelines increases the detection of Helicobacter gastritis and intestinal metaplasia in 400738 sets of gastric biopsies.
The diagnostic yield of individual gastric biopsies and the adherence to the Sydney System guidelines in the United States are unknown. To evaluate the yield of different gastric sampling strategies and determine the adherence to the Sydney System guidelines in a nationwide sample of endoscopists. Using a database of biopsy records diagnosed at a single pathology laboratory, we analysed the results of gastric biopsies taken to evaluate gastric inflammatory conditions in patients with no endoscopic lesions. We then stratified the specimens by site of origin and number of mucosal fragments and calculated the relationship between number and origin of biopsy specimens and detection of Helicobacter pylori and intestinal metaplasia. Of 400 738 biopsy sets, 66.0% were submitted as antrum; 17.4% as corpus; 2.6% as cardia; and 24.7% without topographic identifiers. Separate containers with at least two antral and two corpus specimens (Sydney System compliant) were available in 15 645 cases (3.9%). For antrum, corpus, and unspecified sites, each additional tissue fragment was associated with an incremental increase in the yield for both H. pylori and intestinal metaplasia. Sydney System-compliant sets had significantly greater yield than most of the unspecified or single-site sets (14.8% and 6.0% respectively). The incisura angularis, rarely sampled, yielded minimal additional diagnostic information. In the absence of lesions, the acquisition of at least two biopsy specimens from antrum and corpus, essentially following the Sydney System recommendations, is a sensible strategy that guarantees the maximum diagnostic yield for the most common gastric inflammatory conditions.


Clinical studies with a new steroid--fluorometholone.
Thirty-four cases of uveitis, 26 anterior and 8 posterior, showed elevated intraocular pressure in 25 cases during treatment with local steroids as well as systemic steroids. The patients were treated with drugs to reduce the IOP, but this was not accomplished until the local steroids were stopped. Then a good anti-inflammatory response was obtained with fluorometholone, without further increase of the IOP during the whole treatment. Two cases of episcleritis and one case of allergic conjunctivitis that responded very well to local steroids but showed an increase of the intraocular pressure were treated with fluorometholone. The same anti-inflammatory response was obtained without any increase of the IOP. In our small series, a new steroid has been reviewed. No double-blind studies were done, and the series was not well controlled. But this preliminary result shows that fluorometholone can control inflammation in cases of uveitis, episcleritis, and allergic conjunctivitis as well as other steroids, but without increase of the IOP.


Effect of fructans, prebiotics and fibres on the human gut microbiome assessed by 16S rRNA-based approaches: a review.
The inherent and diverse capacity of dietary fibres, nondigestible oligosaccharides (NDOs) and prebiotics to modify the gut microbiota and markedly influence health status of the host has attracted rising interest. Research and collective initiatives to determine the composition and diversity of the human gut microbiota have increased over the past decade due to great advances in high-throughput technologies, particularly the 16S ribosomal RNA (rRNA) sequencing. Here we reviewed the application of 16S rRNA-based molecular technologies, both community wide (sequencing and phylogenetic microarrays) and targeted methodologies (quantitative PCR, fluorescent in situ hybridisation) to study the effect of chicory inulin-type fructans, NDOs and specific added fibres, such as resistant starches, on the human intestinal microbiota. Overall, such technologies facilitated the monitoring of microbiota shifts due to prebiotic/fibre consumption, though there are limited community-wide sequencing studies so far. Molecular studies confirmed the selective bifidogenic effect of fructans and galactooligosaccharides (GOS) in human intervention studies. Fructans only occasionally decreased relative abundance of Bacteroidetes or stimulated other groups. The sequencing studies for various resistant starches, polydextrose and beta-glucan showed broader effects with more and different types of gut microbial species being enhanced, often including phylotypes of Ruminococcaceae. There was substantial variation in terms of magnitude of response and in individual responses to a specific fibre or NDO which may be due to numerous factors, such as initial presence and relative abundance of a microbial type, diet, genetics of the host, and intervention parameters, such as intervention duration and fibre dose. The field will clearly benefit from a more systematic approach that will support defining the impact of prebiotics and fibres on the gut microbiome, identify biomarkers that link gut microbes to health, and address the personalised response of an individual's microbiota to prebiotics and dietary fibres.


The development and validation of a handicap questionnaire for children with a chronic illness.
This paper describes the development and initial psychometric evaluation of the Handicap Scale for Children (HSC). This questionnaire is based on the London Handicap Scale (LHS), a valid and reliable utility instrument for measuring social participation in adults. A multidisciplinary research group was involved in developing the HSC. The questionnaire was tested in 114 children with a chronic disease and 239 healthy children in the 8-18 age range. Relating the Health Utility Index Mark 3 (HUI3) attributes to corresponding HSC scores tested the assumption that a negative health status would lead to participation problems. Questionnaire development resulted in a five-dimension questionnaire: mobility, physical independence, daily activities, social integration and orientation. Each dimension included one item with a six-point response scale. A higher score indicates greater handicap. Feasibility testing with 10 children showed that none of the children experienced difficulties in filling in the questionnaire. Conceptual validity, measured by correlations between the dimensions of the HSC and HUI3, was satisfactory. As expected, moderate correlation coefficients between predefined pairs of HUI and HSC attributes were found; other correlation coefficients were low. Criterion validity was also satisfactory, as shown by large differences between the healthy and the chronically ill group and by several criteria within the chronically ill group. Based on this initial evaluation, the questionnaire seems feasible and valid for use with children in the age range 8-18 years.


Allergenic pollen in the atmosphere of Kayseri, Turkey.
Airborne pollen are important allergens that cause sensitization in allergic rhinoconjunctivitis and asthma. Our aim was to detect the pollen in the atmosphere of Kayseri, to present a pollen calendar, and to detect the allergenic level of these pollen by performing skin tests on patients. Atmospheric pollen were collected by Durham gravimetric samplers in Kayseri between March and November in the years 1996 and 1997. In our study, we observed pollen belonging to 43 different taxa. The total number of pollen per cm2 was found to be 1,330.8 in 1996 and 1,182.5 in 1997. Most of the pollen were from the taxa Pinus, Poaceae, Chenopodiaceae/Amaranthaceae, Cupressaceae, Populus and Quercus in decreasing order. In the skin tests, pollen of the taxa Poaceae and Chenopodiaceae were found to give the most frequent allergic reactions. It was concluded that preparing an airborne pollen calendar could be useful for medical practice. Nevertheless the skin test data did not really correlate with the aerobiologic data, as skin test reactivity is related to the allergenicity of the pollen and not just to ambient exposure.


Impulsive sounds change European seabass swimming patterns: Influence of pulse repetition interval.
Seismic shootings and offshore pile-driving are regularly performed, emitting significant amounts of noise that may negatively affect fish behaviour. The pulse repetition interval (PRI) of these impulsive sounds may vary considerably and influence the behavioural impact and recovery. Here, we tested the effect of four PRIs (0.5-4.0s) on European seabass swimming patterns in an outdoor basin. At the onset of the sound exposures, the fish swam faster and dived deeper in tighter shoals. PRI affected the immediate and delayed behavioural changes but not the recovery time. Our study highlights that (1) the behavioural changes of captive European seabass were consistent with previous indoor and outdoor studies; (2) PRI could influence behavioural impact differentially, which may have management implications; (3) some acoustic metrics, e.g. SELcum, may have limited predictive power to assess the strength of behavioural impacts of noise. Noise impact assessments need to consider the contribution of sound temporal structure.


Synthesis and analgesic effects of 5-[4-(arylpiperazin-1-yl)alkylamino]-4-benzyl-3-methyl-1,2-oxa zin-6-one s.
A series of 5-[4-(arylpiperazin-1-yl)alkylamino]-4-benzyl-3-methyl-1,2-oxaz in-6-ones was synthesized and evaluated for analgesic activity. The structures of these new oxazine derivatives were confirmed by IR, 1H-NMR spectra and by elemental analysis. The three most active compounds, 3c, 3e and 3g possessed significant antinociceptive effects in the phenylbenzoquinone-induced wrigthing test (PBQ-test) in mice, with ED50 values ranging from 19.7 to 68.0 mg/kg i.p. In addition these compounds presented a low toxicity (LD50 > 800 mg/kg i.p.) and did not significantly reduce the spontaneous locomotor activity of mice. They interacted in a synergistic manner with morphine but nevertheless each compound presented its own profile. Thus the analgesic activity of 3c and 3e was naloxone sensitive, suggesting in mu opioidergic mechanism. Otherwise 3c and 3d analgesia was attenuated by oral administration of yohimbine and therefore seemed to be mediated via noradrenergic pathway. Finally, 5-hydroxytryptophan associated to carbidopa only potentiated 3e analgesia, demonstrating an involvement of a serotoninergic mechanism.


Inactive Rhomboid Protein 2 Mediates Intestinal Inflammation by Releasing Tumor Necrosis Factor-α.
Tumor necrosis factor (TNF)-α is a major proinflammatory cytokine that plays a key role in inflammatory bowel disease (IBD). Inactive rhomboid protein 2 (iRhom2) is essential for activating TNF-α-converting enzyme (TACE) in immune cells, which regulates TNF-α release. The aim of the study was to investigate the role of iRhom2 in intestinal inflammation in IBD. The expression of iRhom2 and TACE in lipopolysaccharide (LPS)-stimulated COLO 205 and RAW 264.7 cells was assessed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. The expression of iRhom2 and TACE in the colonic tissue of IBD patients and 2,4,6-trinitrobenzenesulfonic acid solution (TNBS)-treated mice was determined by RT-PCR and immunohistochemistry. To assess the role of iRhom2 in intestinal inflammation, colitis was induced in wild-type and iRhom2-/- mice by the administration of TNBS enema. In LPS-stimulated COLO 205 and RAW 264.7 cells, the mRNA and protein levels of TACE and iRhom2 were upregulated. The expression of TACE and iRhom2 in the colon of the IBD patients and TNBS-treated mice was significantly enhanced. The inflammatory cells that expressed high levels of iRhom2 in the colon were identified as macrophages. Finally, iRhom2 deficiency ameliorated TNBS-induced colitis by inhibiting TNF-α release. iRhom2 has an important role in intestinal inflammation through TNF-α secretion in immune cells, which suggests that iRhom2 could be a novel therapeutic target for IBD.


Closing the birth registration gap for Every newborn facility birth: literature review and qualitative research.
Birth registration is vital to provide legal identity and access to essential services. Worldwide, approximately 166 million children under five years (just under 25%) are unregistered, yet >80% of all births occur in health facilities in most low- and middle-income countries (LMIC). This study, conducted in association with UNICEF, aims to review facility-based birth registration initiatives, and provide recommendations to close the gap between facility birth and birth registration rates in LMIC. A literature review covering published and grey literature was conducted to identify facility-based initiatives to increase birth registration rates. Semi-structured in-depth interviews were conducted by audio-call with six key global stakeholders to identify additional initiatives, and further insights for barriers and enablers to close the gap. Academic databases and grey literature search yielded 21 studies meeting pre-specified inclusion criteria. Nine barriers preventing birth registration were identified and grouped into three themes: health system, governmental, and societal barriers. Facility-based birth registration initiatives resulted in an increase in birth registration rates. Importantly, health promotion within communities also increased demand for birth registration. In-depth interview respondents provided further detail and supported data found in literature review. Synthesis of the literature and stakeholder interviews noted enablers including inter-sectoral collaboration between health sector and civil registration ministries e.g., placing civil registration offices in health facilities or allowing medical doctors to act as registrars. Facility-based birth registration initiatives can increase birth registration rates in LMIC. Initiatives need to address both supply and demand side of birth registration to improve facility-based birth registration rates. A multi-sectoral approach within governments, and alignment with multiple stakeholders is vital.


Artificial or intelligent? Machine learning and medical selection: possibilities and risks.
This article was migrated. The article was marked as recommended. Machine learning approaches form the basis of "artificial intelligence" and have been increasingly applied in health services settings. It has been shown that such approaches may produce more accurate predictions in some contexts, compared to conventional statistical approaches, and may also reduce the costs of decision-making through automation. Nevertheless, there are both general limitations to developing and implementing machine learning approaches that must be borne in mind. To date, relatively little research has been published on the potential for machine learning to support personnel selection. Moreover, there are particular challenges and issues that need to be considered if such methods are to be used to support decision-making in medical selection scenarios. This article describes some of these potential advantages and challenges and presents an illustrative example, based on real-world data, related to the selection of medical undergraduates.


Reliable Generation of Native-Like Decoys Limits Predictive Ability in Fragment-Based Protein Structure Prediction.
Our previous work with fragment-assembly methods has demonstrated specific deficiencies in conformational sampling behaviour that, when addressed through improved sampling algorithms, can lead to more reliable prediction of tertiary protein structure when good fragments are available, and when score values can be relied upon to guide the search to the native basin. In this paper, we present preliminary investigations into two important questions arising from more difficult prediction problems. First, we investigated the extent to which native-like conformational states are generated during multiple runs of our search protocols. We determined that, in cases of difficult prediction, native-like decoys are rarely or never generated. Second, we developed a scheme for decoy retention that balances the objectives of retaining low-scoring structures and retaining conformationally diverse structures sampled during the course of the search. Our method succeeds at retaining more diverse sets of structures, and, for a few targets, more native-like solutions are retained as compared to our original, energy-based retention scheme. However, in general, we found that the rate at which native-like structural states are generated has a much stronger effect on eventual distributions of predictive accuracy in the decoy sets, as compared to the specific decoy retention strategy used. We found that our protocols show differences in their ability to access native-like states for some targets, and this may explain some of the differences in predictive performance seen between these methods. There appears to be an interaction between fragment sets and move operators, which influences the accessibility of native-like structures for given targets. Our results point to clear directions for further improvements in fragment-based methods, which are likely to enable higher accuracy predictions.


Survival in frontotemporal lobar degeneration in a Korean population.
Frontotemporal lobar degeneration (FTLD) can be subdivided into frontotemporal dementia (FTD), FTD combined with motor neuron disease (FTD-MND), semantic dementia (SD), and progressive nonfluent aphasia (PNFA). FTLD has been considered a rare disorder, and its' demographic and survival data have rarely been studied in Asian population. A survival analysis using the Kaplan-Meier method was performed for 121 consecutive patients with clinically diagnosed FTLD who attended the Memory Disorder Clinic at Samsung Medical Center in Seoul, Republic of Korea, between January 1995 and September 2006. The overall median survival from the onset of the first symptom was 9.6 years (95% CI=8.3-10.8 y). The survival was shortest in FTD-MND (3 y) and longest in SD (11.3 y). The median survival time of FTD (9.8 y) was shorter than that of SD and longer than that of FTD-MND and PNFA. The use of the Cox proportional-hazards model to examine the effect of demographics on survival revealed that only age at onset was associated with survival. In general, our data are comparable with those from the Western countries. However, the female proportion was greater across all subtypes of FTLD and the survival of patients with PNFA was shorter than those of other groups.


Evaluation of RNA-binding specificity of aminoglycosides with DNA microarrays.
We have developed methods for using DNA array technology to probe the entire transcriptome to determine the RNA-binding specificity of ligands. Two methods were investigated. In the first method, the RNA-binding aminoglycoside antibiotic tobramycin was covalently linked to magnetic beads. The beads were bound to human liver mRNA and washed, and specifically bound RNA was eluted, amplified, and analyzed with DNA array technology. A small number of genes were found to bind specifically to the tobramycin beads. In the second method, the aminoglycoside ligand was added directly to the array hybridization reaction, and the signal was compared with a control experiment in the absence of ligand. The aminoglycosides were found to interfere with a small percentage of all hybridization events. These methods differ from traditional DNA array experiments in that the readout is a direct measure of the interaction between mRNA and a ligand, rather than an indirect measure of effect on expression. We expect that the results will lead to the discovery of new aminoglycoside-binding RNA motifs and may also have relevance toward understanding and overcoming the side effects observed with these antibiotics in the clinic.


Effects of multiple exposures to pile driving noise on harbor porpoise hearing during simulated flights-An evaluation tool.
Exploitation of renewable energy from offshore wind farms is substantially increasing worldwide. The majority of wind turbines are bottom mounted, causing high levels of impulsive noise during construction. To prevent temporary threshold shifts (TTS) in harbor porpoise hearing, single strike sound exposure levels (SELSS) are restricted in Germany by law to a maximum of 160 dB re 1 μPa2s at a distance of 750 m from the sound source. Underwater recordings of pile driving strikes, recorded during the construction of an offshore wind farm in the German North Sea, were analyzed. Using a simulation approach, it was tested whether a TTS can still be induced under current protective regulations by multiple exposures. The evaluation tool presented here can be easily adjusted for different sound propagation, acoustic signals, or species and enables one to calculate a minimum deterrence distance. Based on this simulation approach, only the combination of SELSS regulation, previous deterrence, and soft start allow harbor porpoises to avoid a TTS from multiple exposures. However, deterrence efficiency has to be monitored.


Chemosensory Sensitivity after Coffee Consumption Is Not Static: Short-Term Effects on Gustatory and Olfactory Sensitivity.
Chemosensory sensitivity has great variation between individuals. This variation complicates the chemosensory diagnostics, as well as the creation of a meal with universally high hedonic value. To ensure accurate characterization of chemosensory function, a common rule of thumb is to avoid food/beverages one hour before chemosensory testing. However, the scientific foundation of this time of fast remains unclear. Furthermore, the role of coffee on immediate chemosensitivity is not known and may have implications for optimization of gastronomy and hedonia. The aim of this study is to investigate the modularity effects of coffee consumption on immediate gustatory and olfactory sensitivity. We included 155 participants. By applying tests for olfactory and gustatory sensitivity before and after coffee intake, we found no changes in olfactory sensitivity, but significantly altered sensitivity for some basic tastants. We repeated our experimental paradigm using decaffeinated coffee and found similar results. Our results demonstrate that coffee (regular and decaffeinated) alters the subsequent perception of taste, specifically by increasing the sensitivity to sweet and decreasing the sensitivity to bitter. Our findings provide the first evidence of how coffee impacts short-term taste sensitivity and consequently the way we sense and perceive food following coffee intake-an important insight in the context of gastronomy, as well as in chemosensory testing procedures.


Duration of postoperative antibiotic therapy following revision for infected knee and hip arthroplasties.
In the period from 1990 to 1998 28 patients (14 males and 14 females) were treated for infected hip (19 cases) and knee (9 cases) arthroplasties. The median age at index operation was 72 y (range 34-82 y) and at revision surgery 74 y (range 36-83 y). The primary diagnosis was osteoarthritis (16 cases), rheumatoid arthritis (5 cases), failed femoral neck fracture (6 cases) and arthritis secondary to congenital hip dislocation (1 case). Nine cases were infected early, i.e. within 3 months postoperatively, and 19 had late infections. The median interval from index operation to diagnosis of the infected arthroplasty was 18 months (range 1-156 months). Postoperatively, all patients received antibiotics based on susceptibility studies of the causative organism according to culture specimens. The erythrocyte sedimentation rate and CRP level were controlled twice a week during hospitalization and antibiotics were continued until normalization of CRP. The indication for the shift from intravenous to oral antibiotics was a rapid fall in CRP. Antibiotic therapy was terminated when CRP was close to or below 10 mg/l and there was evidence of clinical recovery. Based on the screening of ESR and CRP in this study we believe that CRP is a valuable parameter in deciding when to stop antibiotic therapy. However, it is not clear whether a shorter treatment period is effective or not.


Wearable Cardiorespiratory Monitoring Employing a Multimodal Digital Patch Stethoscope: Estimation of ECG, PEP, LVETand Respiration Using a 55 mm Single-Lead ECG and Phonocardiogram.
Cardiovascular diseases are the main cause of death worldwide, with sleep disordered breathing being a further aggravating factor. Respiratory illnesses are the third leading cause of death amongst the noncommunicable diseases. The current COVID-19 pandemic, however, also highlights the impact of communicable respiratory syndromes. In the clinical routine, prolonged postanesthetic respiratory instability worsens the patient outcome. Even though early and continuous, long-term cardiorespiratory monitoring has been proposed or even proven to be beneficial in several situations, implementations thereof are sparse. We employed our recently presented, multimodal patch stethoscope to estimate Einthoven electrocardiogram (ECG) Lead I and II from a single 55 mm ECG lead. Using the stethoscope and ECG subsystems, the pre-ejection period (PEP) and left ventricular ejection time (LVET) were estimated. ECG-derived respiration techniques were used in conjunction with a novel, phonocardiogram-derived respiration approach to extract respiratory parameters. Medical-grade references were the SOMNOmedics SOMNO HDTM and Osypka ICON-CoreTM. In a study including 10 healthy subjects, we analyzed the performances in the supine, lateral, and prone position. Einthoven I and II estimations yielded correlations exceeding 0.97. LVET and PEP estimation errors were 10% and 21%, respectively. Respiratory rates were estimated with mean absolute errors below 1.2 bpm, and the respiratory signal yielded a correlation of 0.66. We conclude that the estimation of ECG, PEP, LVET, and respiratory parameters is feasible using a wearable, multimodal acquisition device and encourage further research in multimodal signal fusion for respiratory signal estimation.


qPrimerDB: a thermodynamics-based gene-specific qPCR primer database for 147 organisms.
Real-time quantitative polymerase chain reaction (qPCR) is one of the most important methods for analyzing the expression patterns of target genes. However, successful qPCR experiments rely heavily on the use of high-quality primers. Various qPCR primer databases have been developed to address this issue, but these databases target only a few important organisms. Here, we developed the qPrimerDB database, founded on an automatic gene-specific qPCR primer design and thermodynamics-based validation workflow. The qPrimerDB database is the most comprehensive qPCR primer database available to date, with a web front-end providing gene-specific and pre-computed primer pairs across 147 important organisms, including human, mouse, zebrafish, yeast, thale cress, rice and maize. In this database, we provide 3331426 of the best primer pairs for each gene, based on primer pair coverage, as well as 47760359 alternative gene-specific primer pairs, which can be conveniently batch downloaded. The specificity and efficiency was validated for qPCR primer pairs for 66 randomly selected genes, in six different organisms, through qPCR assays and gel electrophoresis. The qPrimerDB database represents a valuable, timesaving resource for gene expression analysis. This resource, which will be routinely updated, is publically accessible at http://biodb.swu.edu.cn/qprimerdb.


Minor histone 2A variants and ubiquinated forms in the native H2A:H2B dimer.
Histone octamers from calf thymus were separated into (H3:H4)2 tetramers and H2A:H2B dimers by chromatography through Sephadex G100. The tetramers and dimers were analyzed for variants, ubiquitin adducts, and proteolyzed forms. The minor histone variants H2A.X and H2A.Z were found to be associated with histone H2B as H2A.X:H2B and H2A.Z:H2B dimers, respectively. Ubiquitin adducts of the H2A's and H2B were also present in H2A:H2B dimers.


Beyond the standard clinical rating scales: fine-grained assessment of post-stroke motor functionality using wearable inertial sensors.
Accurate motor function assessment of post-stroke patients plays a critical role in their rehabilitation interventions. In this paper, we propose an approach to use wearable inertial sensing technology to quantitatively evaluate the patients' motor behavior. Different from existing wearable motor function assessment techniques that focus on building mapping functions that correlate sensed movement signals to the standard clinical rating scales, our approach provides a fine-grained assessment by capturing detailed patterns contained in the patients' movements. We collected data on three subjects including two post-stroke patients who have varying degrees of upper extremity hemiparesis. Our experimental results validate our approach and demonstrate that the captured patterns can be used to complement the standard clinical scores to provide fine-grained motor function assessment and help clinicians to track patients' gradual progress during rehabilitation.


Coronary risk factors and myocardial blood flow in patients evaluated for coronary artery disease: a quantitative [15O]H2O PET/CT study.
There has been increasing interest in quantitative myocardial blood flow (MBF) imaging over the last years and it is expected to become a routinely used technique in clinical practice. Positron emission tomography (PET) using [(15)O]H(2)O is the established gold standard for quantification of MBF in vivo. A fundamental issue when performing quantitative MBF imaging is to define the limits of MBF in a clinically suitable population. The aims of the present study were to determine the limits of MBF and to determine the relationship among coronary artery disease (CAD) risk factors, gender and MBF in a predominantly symptomatic patient cohort without significant CAD. A total of 128 patients (mean age 54 ± 10 years, 50 men) with a low to intermediate pretest likelihood of CAD were referred for noninvasive evaluation of CAD using a hybrid PET/computed tomography (PET/CT) scanner. MBF was quantified with [(15)O]H(2)O at rest and during adenosine-induced hyperaemia. Obstructive CAD was excluded in these patients by means of invasive or CT-based coronary angiography. Global average baseline MBF values were 0.91 ± 0.34 and 1.09 ± 0.30 ml·min(-1)·g(-1) (range 0.54-2.35 and 0.59-2.75 ml·min(-1)·g(-1)) in men and women, respectively (p < 0.01). However, no gender-dependent difference in baseline MBF was seen following correction for rate-pressure product (0.98 ± 0.45 and 1.09 ± 0.30 ml·min(-1)·g(-1) in men and women, respectively; p = 0.08). Global average hyperaemic MBF values were 3.44 ± 1.20 ml·min(-1)·g(-1) in the whole study population, and 2.90 ± 0.85 and 3.78 ± 1.27 ml·min(-1)·g(-1) (range 1.52-5.22 and 1.72-8.15 ml·min(-1)·g(-1)) in men and women, respectively (p < 0.001). Multivariate analysis identified male gender, age and body mass index as having an independently negative impact on hyperaemic MBF. Gender, age and body mass index substantially influence reference values and should be corrected for when interpreting hyperaemic MBF values.


"Affect of anaerobiosis on the antibiotic susceptibility of H. influenzae".
Haemophilus influenzae is a human-restricted facultative anaerobe which resides mostly in the oropharynx. The majority of isolates recovered from the throat are unencapsulated commensals (NTHi), but depending on host susceptibility they cause bronchitis, otitis media and on occasion bacteremia and meningitis. Because of the variable oxygen availability in the various niche permitting bacterium replication, the organism must thrive in well oxygenated surfaces, such as pharyngeal epithelium to anoxic environments like the bottom of a Biofilm and in airway mucus. Other reports indicate that H. influenzae use aerobic respiration, anaerobic respiration and fermentation to generate ATP. To gain insight in to the activity of several classes of antibiotics against five well-characterized unencapsulated H. influenzae in room air, in 5% CO2 and under strict anaerobiosis. We also tested for the role of oxidative killing by all cidal antibiotics. In comparison to room air, testing in 5% CO2 had minimal effects on the susceptibility to aminoglycosides, cephalosporins, tetracycline and chloramphenicol: the MIC of rifampin and ciprofloxacin increased eight fold with certain strains in 5% CO2. All antibiotics, except trimethoprim were cidal under both growth conditions. Aminoglycosides remained bactericidal in a strict anaerobic environment, while a reliable MBC was obtained with trimethoprim only under anaerobic conditions. Kinetic analysis of the cidal action of spectinomycin and tetracycline indicated slower killing anaerobically. An oxidative mechanism for aerobic killing could not be demonstrated. We conclude that β-lactams, cephalosporins, macrolides, tetracycline's, aminoglycosides, chloramphenicol, rifampin and ciprofloxacin are bactericidal against five well-characterizes H. influenzae in an aerobic and anaerobic environment. The activity of trimethoprim was increased in anaerobic conditions.


Country progress towards the Millennium Development Goals: adjusting for socioeconomic factors reveals greater progress and new challenges.
The health Millennium Development Goals (4, 5, 6) impose the same ambitious 2015 targets on every country. Few low-income countries are on track to reach them. Some authors have proposed country-specific targets as a more informative method by which countries can measure their progress against their potential. This paper demonstrates a supplementary approach to assess individual country progress that complements the global goals by adjusting for socioeconomic resources and prior time trends. A minimum performance target adjusts for time and national GDP. Fast-track targets, based on best-performing countries' progress within regional and income groups, adjust for health and non-health sector factors known to affect maternal and child health. Measuring by the minimum performance target, 74% and 59% of low- and middle-income countries are on track for reducing child mortality and maternal mortality, respectively, compared with 69% and 22% using global MDGs. Only 20% and 7% of low- and middle-income countries are on track for the child and maternal mortality fast-track targets. Supplementary targets in maternal and child health, adjusted for each country's resources and policy performance can help countries know if they are truly underperforming relative to their potential. Adjusted targets can also flag countries that have surpassed their potential, and open opportunities for learning from success. Partnership for Maternal, Newborn & Child Health and the Alliance for Health Policy and Systems Research, as part of the Success Factors Study on reducing maternal and child mortality.


Clinical Utility of the Risk Analysis Index as a Prospective Frailty Screening Tool within a Multi-practice, Multi-hospital Integrated Healthcare System.
The goal of this project was to first address barriers to implementation of the Risk Analysis Index (RAI) within a large, multi-hospital, integrated healthcare delivery system, and to subsequently demonstrate its utility for identifying at-risk surgical patients. Prior studies demonstrate the validity of the RAI for evaluating preoperative frailty, but they have not demonstrated the feasibility of its implementation within routine clinical practice. Implementation of the RAI as a frailty screening instrument began as a quality improvement initiative at the University of Pittsburgh Medical Center in July 2016. RAI scores were collected within a REDCap survey instrument integrated into the outpatient electronic health record and then linked to information from additional clinical datasets. NSQIP-eligible procedures were queried within 90 days following the RAI, and the association between RAI and postoperative mortality was evaluated using logistic regression and Cox proportional hazards models. Secondary outcomes such as inpatient length of stay and readmissions were also assessed. RAI assessments were completed on 36,261 unique patients presenting to surgical clinics across five hospitals from July 1 to December 31, 2016, and 8,172 of these underwent NSQIP-eligible surgical procedures. The mean RAI score was 23.6 (SD 11.2), the overall 30-day and 180-day mortality after surgery was 0.7% and 2.6%, respectively, and the median time required to collect the RAI was 33 [IQR 23-53] seconds. Overall clinic compliance with the recommendation for RAI assessment increased from 58% in the first month of the study period to 84% in the sixth and final month. RAI score was significantly associated with risk of death (HR=1.099 [95% C.I.: 1.091 - 1.106], p < 0.001). At an RAI cutoff of ≥37, the positive predictive values for 30- and 90-day readmission were 14.8% and 26.2%, respectively, and negative predictive values were 91.6% and 86.4%, respectively. The RAI frailty screening tool can be efficiently implemented within multi-specialty, multi-hospital healthcare systems. In the context of our findings and given the value of the RAI in predicting adverse postoperative outcomes, health systems should consider implementing frailty screening within surgical clinics.


CRISPR off-target detection with DISCOVER-seq.
DISCOVER-seq (discovery of in situ Cas off-targets and verification by sequencing) is a broadly applicable approach for unbiased CRISPR-Cas off-target identification in cells and tissues. It leverages the recruitment of DNA repair factors to double-strand breaks (DSBs) after genome editing with CRISPR nucleases. Here, we describe a detailed experimental protocol and analysis pipeline with which to perform DISCOVER-seq. The principle of this method is to track the precise recruitment of MRE11 to DSBs by chromatin immunoprecipitation followed by next-generation sequencing. A customized open-source bioinformatics pipeline, BLENDER (blunt end finder), then identifies off-target sequences genome wide. DISCOVER-seq is capable of finding and measuring off-targets in primary cells and in situ. The two main advantages of DISCOVER-seq are (i) low false-positive rates because DNA repair enzyme binding is required for genome edits to occur and (ii) its applicability to a wide variety of systems, including patient-derived cells and animal models. The whole protocol, including the analysis, can be completed within 2 weeks.


Display of Clostridium cellulovorans xylose isomerase on the cell surface of Saccharomyces cerevisiae and its direct application to xylose fermentation.
Xylose isomerase (XI) is a key enzyme in the conversion of D-xylose, which is a major component of lignocellulosic biomass, to D-xylulose. Genomic analysis of the bacterium Clostridium cellulovorans revealed the presence of XI-related genes. In this study, XI derived from C. cellulovorans was produced and displayed using the yeast cell-surface display system, and the xylose assimilation and fermentation properties of this XI-displaying yeast were examined. XI-displaying yeast grew well in medium containing xylose as the sole carbon source and directly produced ethanol from xylose under anaerobic conditions.


Trehalose is a chemical attractant in the establishment of coral symbiosis.
Coral reefs have evolved with a crucial symbiosis between photosynthetic dinoflagellates (genus Symbiodinium) and their cnidarian hosts (Scleractinians). Most coral larvae take up Symbiodinium from their environment; however, the earliest steps in this process have been elusive. Here we demonstrate that the disaccharide trehalose may be an important signal from the symbiont to potential larval hosts. Symbiodinium freshly isolated from Fungia scutaria corals constantly released trehalose (but not sucrose, maltose or glucose) into seawater, and released glycerol only in the presence of coral tissue. Spawning Fungia adults increased symbiont number in their immediate area by excreting pellets of Symbiodinium, and when these naturally discharged Symbiodinium were cultured, they also released trehalose. In Y-maze experiments, coral larvae demonstrated chemoattractant and feeding behaviors only towards a chamber with trehalose or glycerol. Concomitantly, coral larvae and adult tissue, but not symbionts, had significant trehalase enzymatic activities, suggesting the capacity to utilize trehalose. Trehalase activity was developmentally regulated in F. scutaria larvae, rising as the time for symbiont uptake occurs. Consistent with the enzymatic assays, gene finding demonstrated the presence of a trehalase enzyme in the genome of a related coral, Acropora digitifera, and a likely trehalase in the transcriptome of F. scutaria. Taken together, these data suggest that adult F. scutaria seed the reef with Symbiodinium during spawning and the exuded Symbiodinium release trehalose into the environment, which acts as a chemoattractant for F. scutaria larvae and as an initiator of feeding behavior- the first stages toward establishing the coral-Symbiodinium relationship. Because trehalose is a fixed carbon compound, this cue would accurately demonstrate to the cnidarian larvae the photosynthetic ability of the potential symbiont in the ambient environment. To our knowledge, this is the first report of a chemical cue attracting the motile coral larvae to the symbiont.