Characterisation of a GroEL single-ring mutant that supports growth of Escherichia coli and has GroES-dependent ATPase activity.
Binding and folding of substrate proteins by the molecular chaperone GroEL alternates between its two seven-membered rings in an ATP-regulated manner. The association of ATP and GroES to a polypeptide-bound ring of GroEL encapsulates the folding proteins in the central cavity of that ring (cis ring) and allows it to fold in a protected environment where the risk of aggregation is reduced. ATP hydrolysis in the cis ring changes the potentials within the system such that ATP binding to the opposite (trans) ring triggers the release of all ligands from the cis ring of GroEL through a complex network of allosteric communication between the rings. Inter-ring allosteric communication thus appears indispensable for the function of GroEL, and an engineered single-ring version (SR1) cannot substitute for GroEL in vivo. We describe here the isolation and characterisation of an active single-ring form of the GroEL protein (SR-A92T), which has an exceptionally low ATPase activity that is strongly stimulated by the addition of GroES. Dissection of the kinetic pathway of the ATP-induced structural changes in this active single ring can be explained by the fact that the mutation effectively blocks progression through the full allosteric pathway of the GroEL reaction cycle, thus trapping an early allosteric intermediate. Addition of GroES is able to overcome this block by binding this intermediate and pulling the allosteric pathway to completion via mass action, explaining how bacterial cells expressing this protein as their only chaperonin are viable.


'Relief of oppression': an organizing principle for researchers' obligations to participants in observational studies in the developing world.
A central question in the debate about exploitation in international research is whether investigators and sponsors from high-income countries (HIC) have obligations to address background conditions of injustice in the communities in which they conduct their research, beyond the healthcare and other research-related needs of participants, to aspects of their basic life circumstances. In this paper, we describe the Majengo sexually transmitted disease (STD) Cohort study, a long-term prospective, observational cohort of sex workers in Nairobi, Kenya. Despite important scientific contributions and a wide range of benefits to the women of the cohort, most of the women have remained in the sex trade during their long-standing participation in the cohort, prompting allegations of exploitation. The Majengo STD cohort case extends the debate about justice in international research ethics beyond clinical trials into long-term observational research. We sketch the basic features of a new approach to understanding and operationalizing obligations of observational researchers, which we call 'relief of oppression'. 'Relief of oppression' is an organizing principle, analogous to the principle of harm reduction that is now widely applied in public health practice. Relief of oppression aims to help observational researchers working in conditions of injustice and deprivation to clarify their ethical obligations to participants. It aims to bridge the gap between a narrow, transaction-oriented account of avoiding exploitation and a broad account emphasizing obligations of reparation for historic injustices. We propose that relief of oppression might focus researchers' consideration of benefits on those that have some relevance to background conditions of injustice, and so elevate the priority of these benefits, in relation to others that might be considered and negotiated with participants, according to the degree to which the participating communities are constrained in their realization of fundamental freedoms. The over-arching aim of relief of oppression is that, within the range of benefits negotiated over time with the local communities and organizations, an increasing proportion reflects a shared interest in improving participants' fundamental freedoms. We describe how harm reduction serves as a useful analogy for how we envision relief of oppression functioning in international research.


Mitotic stability of infection-induced resistance to plum pox potyvirus associated with transgene silencing and DNA methylation.
Plum pox potyvirus (PPV) infection of transgenic Nicotiana benthamiana plants that expressed the PPV NIb RNA replicase carrying a Gly to Val mutation at the GDD motif (NIbV lines) induced a phenotype of virus resistance and transgene silencing, which was not transmissible to the progeny after self-fertilization (H. S. Guo and J. A. García, Mol. Plant-Microbe Interact. 10:160-170, 1997). Here, we demonstrate that the induced resistance of NIbV plants is mitotically stable after plant propagation by grafting and by in vitro regeneration. Virus replication or residual virus RNA seem not to be required to maintain transgene silencing and virus resistance. Analysis by PCR (polymerase chain reaction) amplification after treatment with methylation-sensitive restriction nucleases indicates that DNA methylation is associated with establishment and maintenance of transgene silencing and virus resistance. Restoration of transgene activity and susceptibility to PPV in sexual progeny correlated with resetting of transgene DNA methylation. On the basis of these and other published results, we present a general model for post-transcriptional gene silencing in which RNA signals, generated either by a silenced nuclear gene or by virus replication, both activate a specific cytoplasmic RNA degradation pathway and induce changes (in particular, DNA methylation) in homologous nuclear genes that switch them from an active to a silenced status.


Structure and electronic properties of in situ synthesized single-layer MoS2 on a gold surface.
When transition metal sulfides such as MoS2 are present in the single-layer form, the electronic properties change in fundamental ways, enabling them to be used, e.g., in two-dimensional semiconductor electronics, optoelectronics, and light harvesting. The change is related to a subtle modification of the band structure due to confinement in the direction perpendicular to the sheets, and there is a considerable interest in understanding how this modification can be controlled and adjusted to generate 2D-materials with functional properties. In this article we report a synthesis procedure together with scanning tunneling microscopy and X-ray photoelectron spectroscopy characterization of two-dimensional single-layer islands of MoS2 synthesized directly on a gold single crystal substrate. Thanks to a periodic modulation of the atom stacking induced by the lattice mismatch, we observe a structural buckling of the MoS2 layer resulting in a characteristic moiré pattern. X-ray photoelectron spectroscopy indicates that the system develops the characteristics of n-doped MoS2 due to electron donation. Scanning tunneling spectroscopy furthermore reflects a convolution of MoS2 and Au donor states where the MoS2 band structure appears modified at the band gap edges. This electronic effect is further modulated by the moiré periodicity and leads to small substrate-induced electronic perturbations near the conduction band minimum in the band gap of MoS2. The results may be highly relevant in the context of nanopatterned two-dimensional materials on metal surfaces, and we propose the MoS2/Au system in this article as a promising candidate to further explore the properties of supported 2D transition-metal dichalcogenides.


Gardening is beneficial for health: A meta-analysis.
There is increasing evidence that gardening provides substantial human health benefits. However, no formal statistical assessment has been conducted to test this assertion. Here, we present the results of a meta-analysis of research examining the effects of gardening, including horticultural therapy, on health. We performed a literature search to collect studies that compared health outcomes in control (before participating in gardening or non-gardeners) and treatment groups (after participating in gardening or gardeners) in January 2016. The mean difference in health outcomes between the two groups was calculated for each study, and then the weighted effect size determined both across all and sets of subgroup studies. Twenty-two case studies (published after 2001) were included in the meta-analysis, which comprised 76 comparisons between control and treatment groups. Most studies came from the United States, followed by Europe, Asia, and the Middle East. Studies reported a wide range of health outcomes, such as reductions in depression, anxiety, and body mass index, as well as increases in life satisfaction, quality of life, and sense of community. Meta-analytic estimates showed a significant positive effect of gardening on the health outcomes both for all and sets of subgroup studies, whilst effect sizes differed among eight subgroups. Although Egger's test indicated the presence of publication bias, significant positive effects of gardening remained after adjusting for this using trim and fill analysis. This study has provided robust evidence for the positive effects of gardening on health. A regular dose of gardening can improve public health.


The effects of attentional focus instructions on punching velocity and impact forces among trained combat athletes.
Research indicates that instructing athlete's to focus on bodily movements (internal focus of attention [IFA]) may hinder performance, whereas instructing them to focus on the movement outcome (external focus of attention [EFA]) often enhances performance. Despite the importance of instructions in striking combat sports, limited research has examined the influence of IFA and EFA on performance in well-trained combat athletes. This study investigated the effects of different instructional cues on punching velocity (m · s-1) and normalised impact forces (N · kg-1) among intermediate (n = 8) and expert (n = 7) competitive boxers and kickboxers. Athletes completed three rounds of 12 maximal effort punches delivered to a punching integrator on three separate days. Day one was a familiarisation session with only control instructions provided. In the following two days athletes randomly received IFA, EFA or control instructions prior to each of the three rounds. Athletes punching with EFA were 4% faster and 5% more forceful than IFA (P < 0.05), and 2% faster and 3% more forceful than control (P < 0.05). Furthermore, experts punched 11% faster and with 13% greater force compared with intermediate athletes (P < 0.05). EFA led to a positive effect on punching performance and should be favoured over IFA and control instructions.


A mating-induced protein of Phytophthora infestans is a member of a family of elicitors with divergent structures and stage-specific patterns of expression.
Five members of an elicitor-like gene family from Phytophthora infestans were examined. The family was identified through the analysis of M81, a mating-induced gene. The predicted M81 product resembled a 42-kDa P. sojae glycoprotein known to elicit defense reactions in plants, including a host of P. infestans, potato. M81 was the most structurally and functionally divergent of the P. infestans genes compared with the P. sojae sequence. M81 lacked elicitor activity, had the lowest protein identity (47%), displayed mating-specific transcription, and had a novel C-terminal domain. The latter contained a 30-residue proline- and threonine-rich motif, which, remarkably, was tandemly repeated 24 to 36 times in different alleles. M81C, M81D, and M81E better resembled the P. sojae protein based on amino acid identity (63 to 75%) and conserved elicitor activity. M81C and M81D mRNA accumulated only during zoosporogenesis, while M81E expression was restricted to hyphae. M81B, an apparent pseudogene, was physically linked to M81. The protein products of each gene were predicted to be extracellular transglutaminases ranging in size from 436 to 1,607 amino acids. Genes with an elicitor, proline- and threonine-rich repeat, and both elicitor and repeat domains were widely distributed throughout Phytophthora infestans. These findings help explain the natural functions of elicitors in pathogen biology and plant-microbe interactions.


Plant-available soil nutrients have a limited influence on cone production patterns of individual white spruce trees.
The resource budget model for mast seeding hypothesizes that soil nutrients proximately influence reproduction. Plants in high soil nutrient (particularly N) areas are predicted to have lower reproductive variability over time and higher mean reproduction. While often examined theoretically, there are relatively few empirical tests of this hypothesis. We quantified cone production of 110 individual white spruce (Picea glauca) trees over seven years and quantified plant-available soil macronutrients (N, Ca, K, Mg, P, S) in natural forest conditions across three years with different cone crop conditions. Each of these plant-available soil nutrients were correlated across years (rs = 0.55-0.89; all > 0.81 for total-N); spatially, total-N availability varied 366-fold across trees. Plant-available soil nutrients did not influence variability or mean annual reproduction, contrary to nutrient perturbation experiments. We examined within-year nutrient and cone-production relationships, and observed significant positive relationships between reproduction and plant-available soil nutrients only in a low-reproduction year preceding a mast event. Both during a mast event and the following year, when overall cone production was very high or very low, there were no relationships. Both external drivers (e.g., weather) and internal resource budgets likely influence soil nutrient-reproduction relationships. These results suggest that plant-available soil nutrients may not be a large factor influencing mast-seeding patterns among individuals in this species.


Exploration of Gender Norms and Socialization Among Early Adolescents: The Use of Qualitative Methods for the Global Early Adolescent Study.
The Global Early Adolescent Study (GEAS) was launched in 2014 with the primary goal of understanding the factors in early adolescence that predispose young people to subsequent sexual risks, and conversely, those that promote healthy sexuality across different cultural contexts. The present article describes the methodology that was used for the first phase of GEAS, which consisted of conducting qualitative research to understand the gendered transitions into adolescence and the role that gender norms play within the key relationships of adolescents. Researchers from each of the sites that had completed data collection were also elicited for their feedback on the key strengths, challenges, and lessons learned from conducting research among 11- to 14-year-old adolescents. The purpose of this article is to present the description of each of the methods that were used in GEAS, as well as the researchers' perspectives of using the methods among early adolescents in their sites. The GEAS is being implemented through a collaboration of university and nongovernmental institutions from 15 cities: Assiut (Egypt) Baltimore (U.S.), Blantyre (Malawi), Cape Town (South Africa), Cochabomba (Bolivia), Cuenca (Ecuador), Edinburgh (Scotland), Ghent (Belgium), Hanoi (Vietnam), Ile-Ife (Nigeria), Kinshasa (DRC), Nairobi (Kenya), New Delhi (India), Ouagadougou (Burkina Faso), and Shanghai (China). Approximately 30 in-depth interviews among adolescents and 30 in-depth interviews with their parent/guardian were conducted at each site, with adults and adolescents interviewed separately. To build trust and increase engagement among the adolescent participants, we used two different visual research methods: (1) timeline exercise which was small group based and (2) the Venn diagram exercise which was conducted individually and used at the start of the in-depth interview. The visual aspects of both the timeline and the Venn diagrams not only helped to produce data for the purposes of the study, but also were a successful way of engaging the adolescent participants across sites. While the narrative interviews produced extremely rich data, researchers did notice that there were a few challenges among the younger adolescents. Challenges were related to the length of the interview, comprehension of questions, as some of the questions were either too abstract or asked adolescents about an experience they had not yet had and therefore could not address or articulate. Conducting the first phase of GEAS revealed important insights for research with participants who are in this developmental phase of early adolescence. Methods that involve greater engagement and those that are visual were shown to work well irrespective of the cultural setting.


How to diagnose sinus fungus balls in the paranasal sinus? An analysis of an institution's cases from January 1999 to December 2006.
The diagnosis of a sinus fungus ball (SFB) is often not clear despite well-defined diagnostic criteria. To study the radiological, intraoperative and histological diagnostic accuracy in comparison to results from mycological and histological analysis. Systematic review of 724 files from patients treated for chronic rhinosinusitis from 1999 - 2006 at our institution. The sensitivity, specificity, positive and negative predictive value (PPV, NPV) of pre- operative CT imaging was 83%, 94%, 56% and 98% respectively, whereas, based on intra- operative findings, it was 98%, 93%, 57% and 100%. A high number of misdiagnoses was found possibly due to sampling error. A severe inflammatory reaction of the surrounding tissue was found more often in SFB than in controls in our study and this we suggest could be an additional sign for fungal infection. Fungal cultures did not contribute to a correct diagnosis.


ESUR/ESUI consensus statements on multi-parametric MRI for the detection of clinically significant prostate cancer: quality requirements for image acquisition, interpretation and radiologists' training.
This study aims to define consensus-based criteria for acquiring and reporting prostate MRI and establishing prerequisites for image quality. A total of 44 leading urologists and urogenital radiologists who are experts in prostate cancer imaging from the European Society of Urogenital Radiology (ESUR) and EAU Section of Urologic Imaging (ESUI) participated in a Delphi consensus process. Panellists completed two rounds of questionnaires with 55 items under three headings: image quality assessment, interpretation and reporting, and radiologists' experience plus training centres. Of 55 questions, 31 were rated for agreement on a 9-point scale, and 24 were multiple-choice or open. For agreement items, there was consensus agreement with an agreement ≥ 70% (score 7-9) and disagreement of ≤ 15% of the panellists. For the other questions, a consensus was considered with ≥ 50% of votes. Twenty-four out of 31 of agreement items and 11/16 of other questions reached consensus. Agreement statements were (1) reporting of image quality should be performed and implemented into clinical practice; (2) for interpretation performance, radiologists should use self-performance tests with histopathology feedback, compare their interpretation with expert-reading and use external performance assessments; and (3) radiologists must attend theoretical and hands-on courses before interpreting prostate MRI. Limitations are that the results are expert opinions and not based on systematic reviews or meta-analyses. There was no consensus on outcomes statements of prostate MRI assessment as quality marker. An ESUR and ESUI expert panel showed high agreement (74%) on issues improving prostate MRI quality. Checking and reporting of image quality are mandatory. Prostate radiologists should attend theoretical and hands-on courses, followed by supervised education, and must perform regular performance assessments. • Multi-parametric MRI in the diagnostic pathway of prostate cancer has a well-established upfront role in the recently updated European Association of Urology guideline and American Urological Association recommendations. • Suboptimal image acquisition and reporting at an individual level will result in clinicians losing confidence in the technique and returning to the (non-MRI) systematic biopsy pathway. Therefore, it is crucial to establish quality criteria for the acquisition and reporting of mpMRI. • To ensure high-quality prostate MRI, experts consider checking and reporting of image quality mandatory. Prostate radiologists must attend theoretical and hands-on courses, followed by supervised education, and must perform regular self- and external performance assessments.


ACT and recovery: integrating evidence-based practice and recovery orientation on assertive community treatment teams.
We examine whether Assertive Community Treatment (ACT), a widely implemented and rigorously studied practice, can successfully incorporate a recovery-oriented approach while continuing to retain program fidelity. We briefly review the effectiveness of ACT as an evidence-based practice, with a focus on adaptations to changing populations and contexts. We explore philosophical similarities and differences between ACT and recovery and examine how fidelity standards, a widely used indicator of how ACT teams operate, support or interfere with the adoption of a recovery-oriented practice. Finally, we provide recommendations on how best to incorporate a recovery orientation into existing ACT teams.


Patients' perceptions of early supported discharge for chronic obstructive pulmonary disease: a qualitative study.
To explore patients' views of an early supported discharge service for chronic obstructive pulmonary disease (COPD). Early supported discharge service (EDS) with discharge at 3.5 days (average length of stay for COPD at the time was 9.5 days). After discharge, patients were visited at home daily for 3 days by a nurse from the early discharge service and then, as required, up to 2 weeks. Purposive, maximum variation sample of 23 mainly older, retired patients admitted to hospital with an acute exacerbation of COPD in 2005. 14 patients listed as receiving EDS, and 9 listed as refusing EDS. Qualitative analysis of audiotaped, semistructured, face-to-face interviews. Setting Economically deprived inner-city borough in England. Results (1) Negotiation and consent. Patients had little recall of being approached to join the scheme. They often felt they had been discharged from hospital before they were ready. They were often unable or unwilling to negotiate timing of discharge with hospital staff. (2) Process of discharge from hospital. Patients experienced difficulties with transport home and supplies of medication. (3) Life at home after a hospital admission. Resuming life at home after an admission for an acute exacerbation for COPD was difficult. Not all patients found the home nursing component of the service helpful. Early supported discharge with domiciliary care is a model that ought to promote a more equal partnership between patient and healthcare, but this did not appear to be the case in practice.


Salvage chemotherapy with cyclophosphamide for recurrent, temozolomide-refractory glioblastoma multiforme.
The primary objective of the current prospective Phase II study of cyclophosphamide (CYC) in adult patients with recurrent, temozolomide-refractory glioblastoma multiforme was to evaluate 6-month progression-free survival (PFS). Forty patients (28 men and 12 women) ages 28-67 years (median age, 51.5 years), with recurrent glioblastoma multiforme were treated. All patients had been treated previously with surgery and involved-field radiotherapy (median dose, 60 grays [Gy]; range, 59-61 Gy). In addition, all patients were treated adjuvantly with either nitrosourea-based chemotherapy (21 patients: procarbazine, lomustine, and vincristine in 13 patients; carmustine in 8 patients) or temozolomide (19 patients). Twenty-one patients who were treated previously with a nitrosourea were treated with temozolomide at the time of first recurrence. Twenty-one patients were treated with CYC at the time of second recurrence, and 19 patients were treated with CYC at the time of first recurrence. CYC was administered intravenously on 2 consecutive days (750 mg/m2 per day) every 4 weeks (operationally defined as a single cycle). Neurologic and neuroradiographic evaluations were performed every 8 weeks. All patients were evaluable. In total, 170 cycles of CYC (median, 2 cycles; range, 2-12 cycles) were administered. CYC-related toxicity included alopecia in all patients (100%), anemia in 6 patients (3.5%), thrombocytopenia in 7 patients (4.1%), and neutropenia in 9 patients (5.3%). Four patients required transfusions (two required red blood cell transfusion, and two required platelet transfusion). One patient developed neutropenic fever without bacteriologic confirmation. No treatment-related deaths occurred. Seven patients (17.5%; 95% confidence interval [95% CI], 8-33%) exhibited a neuroradiographic partial response, 11 patients (27.5%; 95% CI, 15-44%) had stable disease, and 22 patients (55%) had progressive disease after a single cycle of CYC. The time to tumor progression ranged from 2 months to 18 months (median, 2 months). Survival ranged from 3 months to 24 months (median, 4 months). In patients with either a neuroradiographic response or stable disease (n = 18 [45%]), the median time to tumor progression was 6 months (range, 4-18 months; 95% CI, 6-8 months), and the median survival was 10 months (range, 5-24 months; 95% CI, 8-10 months). The 6-month PFS rate was 20%. CYC exhibited modest efficacy with acceptable toxicity in the current cohort of adult patients with recurrent glioblastoma multiforme, all of whom had previously experienced treatment failure after temozolomide chemotherapy.


Recharacterization of ancient DNA miscoding lesions: insights in the era of sequencing-by-synthesis.
Although ancient DNA (aDNA) miscoding lesions have been studied since the earliest days of the field, their nature remains a source of debate. A variety of conflicting hypotheses exist about which miscoding lesions constitute true aDNA damage as opposed to PCR polymerase amplification error. Furthermore, considerable disagreement and speculation exists on which specific damage events underlie observed miscoding lesions. The root of the problem is that it has previously been difficult to assemble sufficient data to test the hypotheses, and near-impossible to accurately determine the specific strand of origin of observed damage events. With the advent of emulsion-based clonal amplification (emPCR) and the sequencing-by-synthesis technology this has changed. In this paper we demonstrate how data produced on the Roche GS20 genome sequencer can determine miscoding lesion strands of origin, and subsequently be interpreted to enable characterization of the aDNA damage behind the observed phenotypes. Through comparative analyses on 390,965 bp of modern chloroplast and 131,474 bp of ancient woolly mammoth GS20 sequence data we conclusively demonstrate that in this sample at least, a permafrost preserved specimen, Type 2 (cytosine-->thymine/guanine-->adenine) miscoding lesions represent the overwhelming majority of damage-derived miscoding lesions. Additionally, we show that an as yet unidentified guanine-->adenine analogue modification, not the conventionally argued cytosine-->uracil deamination, underpins a significant proportion of Type 2 damage. How widespread these implications are for aDNA will become apparent as future studies analyse data recovered from a wider range of substrates.


Transcriptional activation domains of human heat shock factor 1 recruit human SWI/SNF.
Chromatin remodeling complexes such as SWI/SNF use the energy of ATP hydrolysis to remodel nucleosomal DNA and increase transcription of nucleosomal templates. Human heat shock factor one (hHSF1) is a tightly regulated activator that stimulates transcriptional initiation and elongation using different portions of its activation domains. Here we demonstrate that hHSF1 associates with BRG1, the ATPase subunit of human SWI/SNF (hSWI/SNF) at endogenous protein concentrations. We also show that hHSF1 activation domains recruit hSWI/SNF to a chromatin template in a purified system. Mutation of hHSF1 residues responsible for activation of transcriptional elongation has the most severe effect on recruitment of SWI/SNF and association of hHSF1 with BRG1, suggesting that recruitment of chromatin remodeling activity might play a role in stimulation of elongation.


Paraplegia by spinal cord compression as the initial manifestation of Hodgkin's disease: a case report.
The 90% of Hodgkin's disease (HD) cases are originated by lymphnodes whereas 10% by extranodal regions as epidural space. Neurologic complications of HD can be classified as directly resulting from the disease or indirectly originated from the disease or from its treatment. Patients very rarely present with spinal cord compression (SCC) due to epidural HD. Few cases of HD with such presentation have been reported in the literature. Primary spinal extradural HD with no further organ involvement is extremely rare. We report a case of a child with SCC as initial and unique presentation of HD.


A quantitative genomic imbalance gene expression assay in a hexaploid species: wheat (Triticum aestivum).
Responses to allopolyploidy include unequal expression of duplicated genes, gene silencing, and sometimes genomic rearrangements. In plants, the relationship between allelic expression differences arising from changes in regulatory regions and the resulting phenotype is poorly understood because of the complexity of their genomes and lack of efficient methodology to identify regulatory variation. Identifying functionally important regulatory variation in crops such as hexaploid wheat (Triticum aestivum) is in its infancy. More knowledge is required about the contribution of participatory genomes to its transcriptome. In this paper, we demonstrate the use of allelic imbalance assays to quantify relative expression levels across tissues and growth regimes of homoeologous transcripts of the A, B, and D genomes. Polymorphisms in the type I thionins have been used as an example. We show that expression levels vary markedly and interactively over all factors. For this gene, the B genome is the smallest contributor to the transcriptome and the D genome the largest. As additional sequence information is accumulated across genomes, this assay will allow the simple study of relative expression across multiple homeologous loci.


Characteristics of Conversion Disorder in an Urban Academic Children's Medical Center.
Background. Literature on childhood conversion disorder (CD) is sparse and is mostly limited to the outpatient population. Method. Our study retrospectively examines the characteristics of childhood CD in 42 children and adolescents seen by the psychiatric consultation-liaison service in an urban academic medical center with a large minority population. Results. CD accounted for 11% of our consultations. The majority of patients were female adolescents, but in the younger cohort, the male-to-female ratio equalized. Other somatic symptoms and additional psychiatric diagnoses were common, anxiety disorders in particular. Antecedent stressors were identified in 95% of patients, most commonly related to family stressors. Recent or remote history of abuse was rare. Neurological presentations were complex, with almost half of the patients presenting with multiple distinct neurological symptoms. Hyperkinetic symptoms were more common than hypokinetic symptoms, and paroxysmal symptoms were more common than non-paroxysmal. No distress (la belle indifference) was found in only 25% of patients and about half of patients had no socio-academic impairment. High resource utilization was noted based on multiple specialist consultants, diagnostic studies, and length of stay. Conclusion. Much of our data confirms previous findings and contributes to what is becoming a more robust characterization of this population.


Nonisocentric radiosurgical rhizotomy for trigeminal neuralgia.
Although stereotactic radiosurgery is an established procedure for treating trigeminal neuralgia (TN), the likelihood of a prompt and durable complete response is not assured. Moreover, the incidence of facial numbness remains a challenge. To address these limitations, a new, more anatomic radiosurgical procedure was developed that uses the CyberKnife (Accuray, Inc., Sunnyvale, CA) to lesion an elongated segment of the retrogasserian cisternal portion of the trigeminal sensory root. Because the initial experience with this approach resulted in an unacceptably high incidence of facial numbness, a gradual dose and volume de-escalation was performed over several years. In this single-institution prospective study, we evaluated clinical outcomes in a group of TN patients who underwent lesioning with seemingly optimized nonisocentric radiosurgical parameters. Forty-six patients with intractable idiopathic TN were treated between January 2005 and June 2007. Eligible patients were either poor surgical candidates or had failed previous microvascular decompression or destructive procedures. During a single radiosurgical session, a 6-mm segment of the affected nerve was treated with a mean marginal prescription dose of 58.3 Gy and a mean maximal dose of 73.5 Gy. Monthly neurosurgical follow-up was performed until the patient became pain-free. Longer-term follow-up was performed both in the clinic and over the telephone. Outcomes were graded as excellent (pain-free and off medication), good (>90% improvement while still on medication), fair (50-90% improvement), or poor (no change or worse). Facial numbness was assessed using the Barrow Neurological Institute Facial Numbness Scale score. Symptoms disappeared completely in 39 patients (85%) after a mean latency of 5.2 weeks. In most of these patients, pain relief began within the first week. TN recurred in a single patient after a pain-free interval of 7 months; all symptoms abated after a second radiosurgical procedure. Four additional patients underwent a repeat rhizotomy after failing to respond adequately to the first operation. After a mean follow-up period of 14.7 months, patient-reported outcomes were excellent in 33 patients (72%), good in 11 patients (24%), and poor/no improvement in 2 patients (4%). Significant ipsilateral facial numbness (Grade III on the Barrow Neurological Institute Scale) was reported in 7 patients (15%). Optimized nonisocentric CyberKnife parameters for TN treatment resulted in high rates of pain relief and a more acceptable incidence of facial numbness than reported previously. Longer follow-up periods will be required to establish whether or not the durability of symptom relief after lesioning an elongated segment of the trigeminal root is superior to isocentric radiosurgical rhizotomy.


Skewed X-inactivation is common in the general female population.
X-inactivation is a well-established dosage compensation mechanism ensuring that X-chromosomal genes are expressed at comparable levels in males and females. Skewed X-inactivation is often explained by negative selection of one of the alleles. We demonstrate that imbalanced expression of the paternal and maternal X-chromosomes is common in the general population and that the random nature of the X-inactivation mechanism can be sufficient to explain the imbalance. To this end, we analyzed blood-derived RNA and whole-genome sequencing data from 79 female children and their parents from the Genome of the Netherlands project. We calculated the median ratio of the paternal over total counts at all X-chromosomal heterozygous single-nucleotide variants with coverage ≥10. We identified two individuals where the same X-chromosome was inactivated in all cells. Imbalanced expression of the two X-chromosomes (ratios ≤0.35 or ≥0.65) was observed in nearly 50% of the population. The empirically observed skewing is explained by a theoretical model where X-inactivation takes place in an embryonic stage in which eight cells give rise to the hematopoietic compartment. Genes escaping X-inactivation are expressed from both alleles and therefore demonstrate less skewing than inactivated genes. Using this characteristic, we identified three novel escapee genes (SSR4, REPS2, and SEPT6), but did not find support for many previously reported escapee genes in blood. Our collective data suggest that skewed X-inactivation is common in the general population. This may contribute to manifestation of symptoms in carriers of recessive X-linked disorders. We recommend that X-inactivation results should not be used lightly in the interpretation of X-linked variants.


Gender differences on laugh-related traits: A cross-sectional study.
People have different responses to being laughed at, including fear of being laughed at, joy from being laughed at and joy from hearing others laugh. Previous studies have indicated that men have a higher tendency of laughing at others, but there were no gender differences on terms of the other two responses. Socioemotional selectivity theory indicates that individuals pay less attention to negative stimuli as they get older. It is worth investigating whether the gender differences of gelotophobia and gelotophilia occur in certain developmental stages. The present study is a cross-sectional study, with a sample size of 1247 participants; we collected information regarding laugh-related traits among participants 11-66 years old, and we particularly focused on the moderation effect of age on gender differences on gelotophobia, gelotophilia and katagelasticism. The results showed that men had a stronger tendency for laughing at others, but there were no significant gender differences on terms of gelotophobia and gelotophilia; age not only moderated the relationship between gender and gelotophobia but also the relationship between gender and katagelasticism. Male adolescents had higher incidences of gelotophobia and katagelasticism than female adolescents, but this finding was not consistent in the middle adulthood sample. The present study found age to be a moderator between gender and laugh-related traits, and it indicated the influence of age on the fear of being laughed at during early adolescence, but there was no gender difference with respect to katagelasticism in middle adulthood.


Diffusion chamber system for testing of collagen-based cell migration barriers for separation of ligament enthesis zones in tissue-engineered ACL constructs.
A temporary barrier separating scaffold zones seeded with different cell types prevents faster growing cells from overgrowing co-cultured cells within the same construct. This barrier should allow sufficient nutrient diffusion through the scaffold. The aim of this study was to test the effect of two variants of collagen-based barriers on macromolecule diffusion, viability, and the spreading efficiency of primary ligament cells on embroidered scaffolds. Two collagen barriers, a thread consisting of a twisted film tape and a sponge, were integrated into embroidered poly(lactic-co-caprolactone) and polypropylene scaffolds, which had the dimension of lapine anterior cruciate ligaments (ACL). A diffusion chamber system was designed and established to monitor nutrient diffusion using fluorescein isothiocyanate-labeled dextran of different molecular weights (20, 40, 150, 500 kDa). Vitality of primary lapine ACL cells was tested at days 7 and 14 after seeding using fluorescein diacetate and ethidium bromide staining. Cell spreading on the scaffold surface was measured using histomorphometry. Nuclei staining of the cross-sectioned scaffolds revealed the penetration of ligament cells through both barrier types. The diffusion chamber was suitable to characterize the diffusivity of dextran molecules through embroidered scaffolds with or without integrated collagen barriers. The diffusion coefficients were generally significantly lower in scaffolds with barriers compared to those without barriers. No significant differences between diffusion coefficients of both barrier types were detected. Both barriers were cyto-compatible and prevented most of the ACL cells from crossing the barrier, whereby the collagen thread was easier to handle and allowed a higher rate of cell spreading.


Lytic viruses infecting organisms from the three domains of life.
It is probably a biological constant that viruses are found associated with all known life and that they play an essential role in the ecology and evolution of all lifeforms. Lytic viruses have now been discovered that infect organisms from each of the three domains of life. Recently, a new lysis system has been described for two archaeal viruses. This lysis system appears to be novel and distinct from previously described virus-encoded lysis systems. A comparison of lytic viruses from each of the three domains may provide insights concerning the evolution of viruses.


Nickel phytoremediation potential of the Mediterranean Alyssoides utriculata (L.) Medik.
This study investigated the accumulation and distribution of nickel in the leaves and roots of the Mediterranean shrub Alyssoides utriculata to assess its potential use in phytoremediation of Ni contaminated soils. Total (AAS and ICP-MS) Ni, Ca and Mg contents were analyzed in the plants and related to their bioavailability (in EDTA) in serpentine and non-serpentine soils. To find the relationships between the soil available Ni and the Ni content of this species, we also evaluated possible interactions with Ca and Mg. The bioaccumulation factor (BF) and the translocation factor (TF) were determined to assess the tolerance strategies developed by A. utriculata and to evaluate its potential for phytoextraction or phytostabilization. The leaf Ni is higher than 1000 μg g(-1) which categorizes the species as a Ni-hyperaccumulator and a great candidate for Ni-phytoextraction purposes. In addition to the accumulation of Ni, the leaf Mg is also correlated with soil bioavailable concentrations. The Ca uptake and translocation were significantly lower in serpentine plants (higher Ni), as such, the leaf Ca is probably greatly influenced either by the soil's Ni or the soil Ca/Mg ratio. The BFs and TFs are strongly higher than 1 and generally did not significantly differed between plants from serpentine (higher Ni) and non-serpentine soils (lower Ni). The present study highlights for the first time that A. utriculata could be suitable for cleaning Ni-contaminated areas and provides a contribution to the very small volume of data available on the potential use of native Mediterranean plant species from contaminated sites in phytoremediation technologies.


Mapping socio-economic scenarios of land cover change: a GIS method to enable ecosystem service modelling.
We present a GIS method to interpret qualitatively expressed socio-economic scenarios in quantitative map-based terms. (i) We built scenarios using local stakeholders and experts to define how major land cover classes may change under different sets of drivers; (ii) we formalized these as spatially explicit rules, for example agriculture can only occur on certain soil types; (iii) we created a future land cover map which can then be used to model ecosystem services. We illustrate this for carbon storage in the Eastern Arc Mountains of Tanzania using two scenarios: the first based on sustainable development, the second based on 'business as usual' with continued forest-woodland degradation and poor protection of existing forest reserves. Between 2000 and 2025 4% of carbon stocks were lost under the first scenario compared to a loss of 41% of carbon stocks under the second scenario. Quantifying the impacts of differing future scenarios using the method we document here will be important if payments for ecosystem services are to be used to change policy in order to maintain critical ecosystem services.


Marine microgels as a source of cloud condensation nuclei in the high Arctic.
Marine microgels play an important role in regulating ocean basin-scale biogeochemical dynamics. In this paper, we demonstrate that, in the high Arctic, marine gels with unique physicochemical characteristics originate in the organic material produced by ice algae and/or phytoplankton in the surface water. The polymers in this dissolved organic pool assembled faster and with higher microgel yields than at other latitudes. The reversible phase transitions shown by these Arctic marine gels, as a function of pH, dimethylsulfide, and dimethylsulfoniopropionate concentrations, stimulate the gels to attain sizes below 1 μm in diameter. These marine gels were identified with an antibody probe specific toward material from the surface waters, sized, and quantified in airborne aerosol, fog, and cloud water, strongly suggesting that they dominate the available cloud condensation nuclei number population in the high Arctic (north of 80°N) during the summer season. Knowledge about emergent properties of marine gels provides important new insights into the processes controlling cloud formation and radiative forcing, and links the biology at the ocean surface with cloud properties and climate over the central Arctic Ocean and, probably, all oceans.


The role of water, sanitation and hygiene interventions in reducing soil-transmitted helminths: interpreting the evidence and identifying next steps.
The transmission soil transmitted helminths (STH) occurs via ingestion of or contact with infective stages present in soil contaminated with human faeces. It follows therefore that efforts to reduce faecal contamination of the environment should help to reduce risk of parasite exposure and improvements in water, sanitation and hygiene (WASH) are seen as essential for the long-term, sustainable control of STH. However, the link between WASH and STH is not always supported by the available evidence from randomised controlled trials, which report mixed effects of WASH intervention on infection risk. This review critically summarises the available trial evidence and offers an interpretation of the observed heterogeneity in findings. The review also discusses the implications of findings for control programmes and highlights three main issues which merit further consideration: intervention design, exposure assessment, and intervention fidelity assessment.


A novel method of ligand peptidomics to identify peptide ligands binding to AQP2-expressing plasma membranes and intracellular vesicles of rat kidney.
Aquaporin-2 (AQP2), the vasopressin-regulated water channel in collecting duct principal cells, plays a key role in the regulation of body water balance. We aimed to isolate high-affinity peptide ligands that bind to immunoisolated AQP2-expressing plasma membrane (PM) or intracellular vesicle (ICV) preparations from rat kidney by the in vitro phage display technique. Immunoblotting revealed that AQP2 was exclusively expressed in the immunoisolated AQP2 membrane fractions (PM and ICV), compared with the nonimmunoisolated or preimmune IgG pulldown rat kidney samples. Moreover, AQP1 or H+-ATPase (B1 subunit) expression was minimal in the immunoisolated AQP2 membrane fractions, indicating the specificity of AQP2 membrane isolation. A phage peptide library based on T7 415-1b phage vector displaying CX7C was constructed. After three rounds of biopanning, seven phage clones of high frequency were selected, which showed high affinity to the AQP2-containing PM or ICV fractions compared with a nonrecombinant T7 insertless phage clone. In contrast, these phage clones showed lower affinity to H+-ATPase-containing fractions. Fluorescein-conjugated peptide labeling was associated with intracellular compartment and PM of primary cultured inner medullary collecting duct cells, relative to absent or very weak labeling with fluorescein-conjugated control peptide. Library analyses demonstrated proteins that had motifs homologous to the peptide ligands, albeit with a high probability of a random match due to short peptide sequences. In summary, we applied the in vitro phage display technique to identify high-affinity peptide ligands to AQP2-expressing membranes. Library analyses identified proteins having homologous motifs, which need to be examined for involvement in AQP2 trafficking and regulation.


Lack of significant effect of coffee and caffeine on fluoride metabolism in rats.
It has been reported that rat plasma fluoride (F) concentrations are higher by up to 100% when F is administered ig in coffee or a caffeine solution compared with when it is administered in water. It was hypothesized that the consumption of caffeinated beverages has contributed to the prevalence of dental fluorosis. The present studies were done to determine the physiological mechanisms for these effects. For approximately 2 h after F was administered in coffee, plasma F concentrations were higher than when administered in water, decaffeinated coffee, or a caffeine solution (3 mg/kg), but the intergroup differences were small and generally not statistically significant. The 4-hour plasma AUC values did not differ with statistical significance. There were no differences among the groups in the renal or extrarenal (skeletal) clearances of F, which suggested that the higher plasma F concentrations in the coffee groups may have been due to a slight and transient increase in absorption rate. The possibility that caffeine per se might elevate endogenous plasma F and calcium concentrations was excluded after caffeine (25 mg/kg) ig without F was given. In addition, the renal excretion, clearance, and fractional renal clearance of calcium did not differ among the groups. The results indicated that decaffeinated coffee and caffeine had no effect on F metabolism, whereas caffeinated coffee appeared to increase the initial absorption rate but not the 4-hour bio-availability.(ABSTRACT TRUNCATED AT 250 WORDS)


An Integrated View of the Effects of Wine Polyphenols and Their Relevant Metabolites on Gut and Host Health.
Over the last few decades, polyphenols, and flavonoids in particular, have attracted the interest of researchers, as they have been associated with the health-promoting effects derived from diets rich in vegetables and fruits, including moderate wine consumption. Recent scientific evidence suggests that wine polyphenols exert their effects through interactions with the gut microbiota, as they seem to modulate microbiota and, at the same time, are metabolized by intestinal bacteria into specific bioavailable metabolites. Microbial metabolites are better absorbed than their precursors and may be responsible for positive health activities in the digestive system (local effects) and, after being absorbed, in tissues and organs (systemic effects). Differences in gut microbiota composition and functionality among individuals can affect polyphenol activity and, therefore, their health effects. The aim of this review is to integrate the understanding of the metabolism and mechanisms of action of wine polyphenols at both local and systemic levels, underlining their impact on the gut microbiome and the inter-individual variability associated with polyphenols' metabolism and further physiological effects. The advent of promising dietary approaches linked to wine polyphenols beyond the gut microbiota community and metabolism are also discussed.


Vertically porous nickel thin film supported Mn3O4 for enhanced energy storage performance.
Three-dimensionally porous metal materials are often used as the current collectors and support for the active materials of supercapacitors. However, the applications of vertically porous metal materials in supercapacitors are rarely reported, and the effect of vertically porous metal materials on the energy storage performance of supported metal oxides is not explored. To this end, the Mn3O4-vertically porous nickel (VPN) electrodes are fabricated via a template-free method. The Mn3O4-VPN electrode shows much higher volumetric specific capacitances than that of flat nickel film supported Mn3O4 with the same loading under the same measurement conditions. The volumetric specific capacitance of the vertically porous nickel supported Mn3O4 electrode can reach 533Fcm(-3) at the scan rate of 2mVs(-1). The fabricated flexible all-solid microsupercapacitor based on the interdigital Mn3O4-VPN electrode has a volumetric specific capacitance of 110Fcm(-3) at the current density of 20μAcm(-2). The capacitance retention rate of this microsupercapacitor reaches 95% after 5000 cycles under the current density of 20μAcm(-2). The vertical pores in the nickel electrode not only fit the micro/nanofabrication process of the Mn3O4-VPN electrode, but also play an important role in enhancing the capacitive performances of supported Mn3O4 particles.


Parenteral glutamine supplementation does not reduce the risk of mortality or late-onset sepsis in extremely low birth weight infants.
Glutamine is one of the most abundant amino acids in both plasma and human milk, yet it is not included in standard intravenous amino acid solutions. Previous studies have suggested that parenteral nutrition (PN) supplemented with glutamine may reduce sepsis and mortality in critically ill adults. Whether glutamine supplementation would provide a similar benefit to extremely low birth weight (ELBW) infants is not known. We performed a multicenter, randomized, double-masked, clinical trial to assess the safety and efficacy of early PN supplemented with glutamine in decreasing the risk of death or late-onset sepsis in ELBW infants. Infants 401 to 1000 g were randomized within 72 hours of birth to receive either TrophAmine (control) or an isonitrogenous study amino acid solution with 20% glutamine whenever they received PN up to 120 days of age, death, or discharge from the hospital. The primary outcome was death or late-onset sepsis. Of the 721 infants who were assigned to glutamine supplementation, 370 (51%) died or developed late-onset sepsis, as compared with 343 of the 712 infants (48%) assigned to control (relative risk: 1.07; 95% confidence interval: 0.97-1.17). Glutamine had no effect on tolerance of enteral feeds, necrotizing enterocolitis, or growth. No significant adverse events were observed with glutamine supplementation. Parenteral glutamine supplementation as studied did not decrease mortality or the incidence of late-onset sepsis in ELBW infants. Consequently, although no harm was demonstrated, routine use of parenteral glutamine supplementation cannot be recommended in this population.


Management of metabolic effects associated with anticancer agents targeting the PI3K-Akt-mTOR pathway.
Agents inhibiting the phosphoinositide 3-kinase-Akt-mammalian target of rapamycin (PAM) pathway are currently in various stages of clinical development in oncology, ranging from some in early-phase evaluations to others that have already received regulatory approval for treatment in advanced cancers. The administration of PAM pathway inhibitors has been associated with metabolic toxicities of hyperlipidemia and hyperglycemia. The PAM Task Force of the National Cancer Institute Investigational Drug Steering Committee convened an interdisciplinary expert panel to review the pathophysiology of hyperlipidemia and hyperglycemia induced by PAM pathway inhibitors, summarize the incidence of these metabolic toxicities induced by such agents in the current literature, advise on clinical trial screening and monitoring criteria, and provide management guidance and therapeutic goals on occurrence of these toxicities. The overarching aim of this consensus report is to raise awareness of these metabolic adverse events to enable their early recognition, regular monitoring, and timely intervention in clinical trials. Hyperglycemia and hyperlipidemia are generally not acutely toxic and most often reversible with therapeutic intervention. Dose modifications or discontinuation of PAM pathway inhibitors should only be considered in situations of severe events or if progressive metabolic derangement persists after therapeutic interventions have been attempted for a sufficient duration. Specialty consultation should be sought to aid clinical trial planning and the management of these metabolic adverse events.


Baseline correction for stray light in log-ratio diode laser absorption measurements.
Log-ratio detection is a convenient technique for making temperature and concentration measurements using sensors based on tunable diode laser absorption spectroscopy. In many practical sensing applications, it is difficult to avoid stray light falling on the signal photodiode of the sensor. This stray light acts as noncommon-mode interference and introduces a systematic error in absorption measurements, which is not removed by baseline subtraction. This paper analyzes the factors that determine this systematic error and also presents a calibration method that can correct for it. This correction method is verified using a simple experiment.


Effects of Pulse Shape and Polarity on Sensitivity to Cochlear Implant Stimulation: A Chronic Study in Guinea Pigs.
Most cochlear implants (CIs) stimulate the auditory nerve with trains of symmetric biphasic pulses consisting of two phases of opposite polarity. Animal and human studies have shown that both polarities can elicit neural responses. In human CI listeners, studies have shown that at suprathreshold levels, the anodic phase is more effective than the cathodic phase. In contrast, animal studies usually show the opposite trend. Although the reason for this discrepancy remains unclear, computational modelling results have proposed that the degeneration of the peripheral processes of the neurons could lead to a higher efficiency of anodic stimulation. We tested this hypothesis in ten guinea pigs who were deafened with an injection of sysomycin and implanted with a single ball electrode inserted in the first turn of the cochlea. Animals were tested at regular intervals between 1 week after deafening and up to 1 year for some of them. Our hypothesis was that if the effect of polarity is determined by the presence or absence of peripheral processes, the difference in polarity efficiency should change over time because of a progressive neural degeneration. Stimuli consisted of charge-balanced symmetric and asymmetric pulses allowing us to observe the response to each polarity individually. For all stimuli, the inferior colliculus evoked potential was measured. Results show that the cathodic phase was more effective than the anodic phase and that this remained so even several months after deafening. This suggests that neural degeneration cannot entirely account for the higher efficiency of anodic stimulation observed in human CI listeners.


Measuring Children's Media Use in the Digital Age: Issues and Challenges.
In this new and rapidly changing era of digital technology, there is increasing consensus among media scholars that there is an urgent need to develop measurement approaches which more adequately capture media use The overarching goal of this paper is facilitate the development of measurement approaches appropriate for capturing children's media use in the digital age. The paper outlines various approaches to measurement, focusing mainly on those which have figured prominently in major existing studies of children's media use. We identify issues related to each technique, including advantages and disadvantages. We also include a review of existing empirical comparisons of various methodologies. The paper is intended to foster discussion of the best ways to further research and knowledge regarding the impact of media on children.


Identification of children at risk of influenza-related complications in primary and ambulatory care: a systematic review and meta-analysis.
Interventions to prevent influenza-related complications are recommended for individuals at the greatest risk of serious clinical deterioration. However, guidelines are based on consensus opinion rather than evidence, and do not specify risk factors in children. We aimed to provide an evidence-based definition of children who are most at risk of such complications. In this systematic review, we searched the Medline and Medline In Process, Embase, Science Citation Index, and CINAHL databases for studies published between inception and April 3, 2013. We included studies that reported data for underlying disorders and complications in children presenting in primary or ambulatory care with influenza or influenza-like illness. We requested unpublished data from investigators of studies that had obtained, but not published, relevant data. We analysed data with univariable meta-analysis and individual patient data multivariable meta-analysis methods. The primary outcome was admission to hospital as a proxy for complications of influenza or influenza-like illness. We included 28 articles that reported data from 27 studies (14 086 children). Strong risk factors for hospital admission were neurological disorders (univariable odds ratio [OR] 4· 62, 95% CI 2·82-7·55), prematurity (4·33, 2·47-7·58), sickle cell disease (3·46, 1·63-7·37), immunosuppression (2·39, 1·24-4·61), diabetes (2·34, 1·20-4·58), and age younger than 2 years (2·51, 1·71-3·69). However, reactive airways disease including asthma (1·36, 0·82-2·26) and obesity (0·99, 0·61-1·62) were not found to be risk factors. On the basis of individual patient data multivariable analysis (1612 children, four studies), the risk of hospital admission was higher in children with more than one risk factor than in children with just one risk factor, when age younger than 2 years was included as a risk factor (92 [74%] of 124 vs 428 [52%] of 817; difference 22%, 95% CI 13-30%, p<0·0001). We identified prematurity as a new strong risk factor for influenza-related complications in children. Our findings also support the inclusion of neurological disorders, sickle cell disease, immunosuppression, diabetes, and age younger than 2 years as risk factors in existing guidelines. Interventions to prevent influenza-related complications should be prioritised in these groups, but should also be considered for other children, especially those with more than one risk factor or severe underlying comorbidities. UK National Institute for Health Research.


A universal method to produce low-work function electrodes for organic electronics.
Organic and printed electronics technologies require conductors with a work function that is sufficiently low to facilitate the transport of electrons in and out of various optoelectronic devices. We show that surface modifiers based on polymers containing simple aliphatic amine groups substantially reduce the work function of conductors including metals, transparent conductive metal oxides, conducting polymers, and graphene. The reduction arises from physisorption of the neutral polymer, which turns the modified conductors into efficient electron-selective electrodes in organic optoelectronic devices. These polymer surface modifiers are processed in air from solution, providing an appealing alternative to chemically reactive low-work function metals. Their use can pave the way to simplified manufacturing of low-cost and large-area organic electronic technologies.


Inflammosome in the human endometrium: further step in the evaluation of the "maternal side".
To investigate the expression of inflammosome components (NALP-3, associated speck-like protein containing a CARD [ASC]) and their activation (caspase-1, interleukin [IL]-1β, and IL-18 secretion) in the human endometrium from fertile and women with history of recurrent pregnancy loss (RPL). Experimental study. University hospital. Ten fertile women (control group [CTR]) and 30 women with RPL. None. Endometrial samples were collected by hysteroscopy during the putative window of implantation and evaluated for chronic endometrial inflammation by hystopathological analysis. Inflammosome expression was analysed by immunohystochemical staining (27 RPL and 10 CTR women). The expression of NALP-3 and ASC protein was quantified by Western blot (30 RPL and 10 CTR women). Caspase-1 activation and IL-1β and IL-18 secretion was quantified by ELISA (30 RPL and 10 CTR women). We observed a significantly increased expression of inflammasome NALP-3 and ASC protein, an increased activation of caspase-1, and increased levels of IL-1β and IL-18 in RPL endometrium compared with CTR. Abnormal activation of endometrial innate immunity by means of inflammosome, stimulated by pathogen- or damage-associated molecular patterns, may represent an additional mechanism, currently not investigated, negatively interfering with endometrial receptivity. More studies are required [1] to identify the primary trigger of endometrial inflammosome activation and its clinical impact in the occurrence of RPL; and [2] to validate the inflammosome components as a novel family of endometrial biomarkers and promising therapeutic targets in RPL.


Effect of hyperoxia on oxygen consumption in exercising ponies.
Published reports of oxygen consumption (VO2) during exercise in hyperoxia are equivocal. By and large, when measured at the lung using respiratory gas equations, VO2 is elevated in hyperoxia and, when measured at the blood-tissue level using the cardiovascular Fick (CVF) equation, it is unchanged. We sought to provide some insight into this problem by making through the use of both equations simultaneous determinations of VO2 during hyperoxia in exercising ponies. In normoxia, during treadmill exercise (115 m/min, 10% grade) of seven ponies, there was no difference in exercise VO2, whether it was measured by the Haldane transformation (HT) or CVF equations (P greater than 0.05). In hyperoxia, the exercise VO2 was significantly increased from the normoxia condition (P less than 0.05) when measured by the HT equation but not when measured by the CVF equation (P greater than 0.05). By use of the CVF equation as the method of choice for VO2 determinations in hyperoxia, the present data show no change in exercise VO2 in the hyperoxic condition.


Posterior tibial nerve stimulation as neuromodulative treatment of lower urinary tract dysfunction.
Recently, intermittent percutaneous posterior tibial nerve stimulation was introduced as a treatment modality filling the gap between conservative and surgical therapies in patients with certain types of lower urinary tract dysfunction. In a prospective multicenter trial posterior tibial nerve stimulation was evaluated in 37 patients who presented with symptoms of bladder overactivity, that is the urgency and frequency syndrome and/or urge incontinence, and 12 with nonobstructive urinary retention. Results were recorded in voiding diaries and on quality of life questionnaires before and after treatment. Patients were classified as responders, including those in whom therapy was successful and chose to continue treatment after the initial 12 weeks, and nonresponders, those who chose to stop treatment. Overall, a positive response was seen in 60% of all patients. In patients with bladder overactivity a statistically significant decrease was observed in leakage episodes, number of pads used, voiding frequency and nocturia, and an equal increase in mean and smallest volume voided. Improvements were also seen in nonobstructive urinary retention, including number of catheterizations, total and mean volume catheterized, and total and mean volume voided. Disease specific quality of life and some domains of general quality of life improved, especially of bladder overactivity. Only mild side effects were observed. Posterior tibial nerve stimulation is a minimally invasive and successful treatment option for patients with certain types of lower urinary tract dysfunction.


Texture segmentation influences the spatial profile of presaccadic attention.
Attention is important for selecting targets for action. Several studies have shown that attentional selection precedes eye movements to a target, and results in an enhanced sensitivity at the saccade goal. Typically these studies have used isolated targets on blank backgrounds, which are rare in real-world situations. Here, we examine the spatial profile of sensitivity around a saccade target on a textured background and how the influence of the surrounding context develops over time. We used two textured backgrounds: a uniform texture, and a concentric arrangement of an inner and an outer texture with orthogonal orientations. For comparison, we also measured sensitivity around the target on a blank background. The spatial profile of sensitivity was measured with a brief, dim, probe flashed around the saccade target. When the target was on a blank or a uniformly textured background, spatial sensitivity peaked near the target location around 350 ms after cue onset and declined with distance from the target. However, when the background was made up of an inner and outer texture, sensitivity to the inner texture was uniformly high, peaking at about 350 ms after cue onset, suggesting that the entire inner texture was selected along with the target. The enhancement of sensitivity on the inner texture was much smaller when observers attended the target covertly and performed the probe-detection task. Thus, our results suggest that the surface representation around the target is taken into account when an observer actively plans to interact with the target.


Biology of subjective cognitive complaint amongst geriatric patients: vitamin D involvement.
Insufficiency in vitamin D, a neurosteroid hormone, is associated with cognitive decline in older adults. The impact on the subjective perception of cognitive decline has not yet been examined. The objective of this cross-sectional hospital-based study was to determine whether vitamin D insufficiency was associated with subjective cognitive complaint amongst geriatric patients. Ninety-nine consecutive Caucasian in- and outpatients recruited in the 'Cognition and LIPophilic vitamins' (CLIP) study, who had no advanced cognitive disorders (i.e., Mini-Mental State Examination score≥20) and who took no vitamin D supplements, were categorized into 2 groups based on vitamin D insufficiency (i.e., serum 25-hydroxyvitamin D≥75 nmol/L). Subjective cognitive complaint was examined using the Memory Complaint Questionnaire (MAC-Q; score 0-30, best). MAC-Q score<15 out of 30 defined severe cognitive complaint. Age, gender, body mass index, education level, comorbidity burden, functional autonomy, mood, and serum concentrations of parathyroid hormone, calcium, thyroid-stimulating hormone and vitamin B12, and estimated glomerular filtration rate were used as potential confounders. Compared to participants with serum 25OHD>75 nmol/L, those with vitamin D insufficiency (n=89) had a lower mean MAC-Q score (14.9±2.9 versus 17.1±1.6, P=0.02) and more often a MAC-Q score<15 (52.8% versus 10.0%, P=0.01). Vitamin D insufficiency was inversely associated with the MAC-Q score (adjusted β=-2.84, P=0.03), and positively associated with severe cognitive complaint (adjusted OR=10.07, P=0.03). Vitamin D insufficiency was associated with subjective cognitive complaint in the studied cohort of geriatric patients.


Pheromonal induction of spatial learning in mice.
Many mammals use scent marking for sexual and competitive advertisement, but little is known about the mechanism by which scents are used to locate mates and competitors. We show that darcin, an involatile protein sex pheromone in male mouse urine, can rapidly condition preference for its remembered location among females and competitor males so that animals prefer to spend time in the site even when scent is absent. Learned spatial preference is conditioned through contact with darcin in a single trial and remembered for approximately 14 days. This pheromone-induced learning allows animals to relocate sites of particular social relevance and provides proof that pheromones such as darcin can be highly potent stimuli for social learning.


The effectiveness of pre-operative exercise physiotherapy rehabilitation on the outcomes of treatment following anterior cruciate ligament injury: a systematic review.
To evaluate the effectiveness of pre-operative exercise physiotherapy rehabilitation on the outcomes of treatment following anterior cruciate ligament injury. The following databases were searched: PubMed, Ovid, The Cochrane Library and Web of Science. Studies published between the inception of the databases and December 2015 were sought using appropriate keywords in various combinations. This search was supplemented with a manual search of the references of selected studies. Studies were assessed for methodological quality using the Physiotherapy Evidence Database scale. A total of 500 studies were identified, of which eight studies met the inclusion criteria and were included in the present review. The average Physiotherapy Evidence Database score for the studies included was 5.8, which reflects an overall moderate methodological quality. The eight studies investigated a total of 451 subjects of which 71% ( n=319) were males. The age of the participants in the eight studies ranged from 15 to 57 years. The duration of the intervention in the studies ranged from 3 to 24 weeks. This review found that pre-operative physiotherapy rehabilitation is effective for improving the outcomes of treatment following anterior cruciate ligament injury, including increasing knee-related function and improving muscle strength. However, whilst there was a significant improvement in quality of life from baseline following intervention, no significant difference in quality of life was found between the control and intervention groups. There is evidence to suggest that pre-operative physiotherapy rehabilitation is beneficial to patients with anterior cruciate ligament injury.


A Novel Approach to Preoperative Risk Stratification in Endometrial Cancer: The Added Value of Immunohistochemical Markers.
Background: The current model used to preoperatively stratify endometrial cancer (EC) patients into low- and high-risk groups is based on histotype, grade, and imaging method and is not optimal. Our study aims to prove whether a new model incorporating immunohistochemical markers, L1CAM, ER, PR, p53, obtained from preoperative biopsy could help refine stratification and thus the choice of adequate surgical extent and appropriate adjuvant treatment. Materials and Methods: The following data were prospectively collected from patients operated for EC from January 2016 through August 2018: age, pre- and post-operative histology, grade, lymphovascular space invasion, L1CAM, ER, PR, p53, imaging parameters obtained from ultrasound, CT chest/abdomen, final FIGO stage, and current decision model (based on histology, grade, imaging method). Results: In total, 132 patients were enrolled. The current model revealed 48% sensitivity and 89% specificity for high-risk group determination. In myometrial invasion >50%, lower levels of ER (p = 0.024), PR (0.048), and higher levels of L1CAM (p = 0.001) were observed; in cervical involvement a higher expression of L1CAM (p = 0.001), lower PR (p = 0.014); in tumors with positive LVSI, higher L1CAM (p = 0.014); in cases with positive LN, lower expression of ER/PR (p < 0.001), higher L1CAM (p = 0.002) and frequent mutation of p53 (p = 0.008). Cut-offs for determination of high-risk tumors were established: ER <78% (p = 0.001), PR <88% (p = 0.008), and L1CAM ≥4% (p < 0.001). The positive predictive values (PPV) for ER, PR, and L1CAM were 87% (60.8-96.5%), 63% (52.1-72.8%), 83% (70.5-90.8%); the negative predictive values (NPV) for each marker were as follows: 59% (54.5-63.4%), 65% (55.6-74.0%), and 77% (67.3-84.2%). Mutation of p53 revealed PPV 94% (67.4-99.1%) and NPV 61% (56.1-66.3%). When immunohistochemical markers were included into the current diagnostic model, sensitivity improved (48.4 vs. 75.8%, p < 0.001). PPV was similar for both methods, while NPV (i.e., the probability of extremely low risk in negative test cases) was improved (66 vs. 78.9%, p < 0.001). Conclusion: We proved superiority of new proposed model using immunohistochemical markers over standard clinical practice and that new proposed model increases accuracy of prognosis prediction. We propose wider implementation and validation of the proposed model.


Urbanization affects peak timing, prevalence, and bimodality of influenza pandemics in Australia: Results of a census-calibrated model.
We examine salient trends of influenza pandemics in Australia, a rapidly urbanizing nation. To do so, we implement state-of-the-art influenza transmission and progression models within a large-scale stochastic computer simulation, generated using comprehensive Australian census datasets from 2006, 2011, and 2016. Our results offer a simulation-based investigation of a population's sensitivity to pandemics across multiple historical time points and highlight three notable trends in pandemic patterns over the years: increased peak prevalence, faster spreading rates, and decreasing spatiotemporal bimodality. We attribute these pandemic trends to increases in two key quantities indicative of urbanization: the population fraction residing in major cities and international air traffic. In addition, we identify features of the pandemic's geographic spread that we attribute to changes in the commuter mobility network. The generic nature of our model and the ubiquity of urbanization trends around the world make it likely for our results to be applicable in other rapidly urbanizing nations.


Giant vesicles from rehydrated crude mixtures containing unexpected mixtures of amphiphiles formed under plausibly prebiotic conditions.
Giant lipid vesicles resemble compartments of biological cells, mimicking them in their dimension, membrane structure and partly in their membrane composition. The spontanenous appearance of closed membranes composed of bilayers of self-assembling amphiphiles was likely a prerequisite for Darwinian competitive behavior to set in at the molecular level. Such compartments should be dynamic in their membrane composition (evolvable), and sufficiently stable to harbor macromolecules (leak-free), yet semi-permeable for reactive small molecules to get across the membrane (stay away from chemical equilibrium). Here we describe bottom-up experiments simulating prebiotic environments that support the formation of simple amphiphilic molecules capable of self-assembling into vesicular objects on the micrometer scale. Long-chain alkyl phosphates, together with related amphiphilic compounds, were formed under simulated prebiotic phosphorylation conditions by using cyanamide, a recognized prebiotic chemical activator and a precursor for several compound classes. Crude dry material of the thus obtained prebiotic mixtures formed multilamellar giant vesicles once rehydrated at the appropriate pH and in the presence of plausibly prebiotic co-surfactants, as observed by optical microscopy. The size and the shape of lipid aggregates tentatively suggest that prebiotic lipid assemblies could encapsulate peptides or nucleic acids that could be formed under similar chemical prebiotic conditions. The formation of prebiotic amphiphiles was monitored by using TLC, IR, NMR and ESI-MS and UPLC-HRMS. In addition we provide a spectroscopic analysis of cyanamide under simulated prebiotic conditions in the presence of phosphate sources and spectroscopic analysis of O-phosphorylethanolamine as a plausible precursor for phosphoethanolamine lipids.


Combining the endoplasmic reticulum stress-inducing agents bortezomib and fenretinide as a novel therapeutic strategy for metastatic melanoma.
Single-agent chemotherapy is largely the treatment of choice for systemic therapy of metastatic melanoma, but survival rates are low, and novel adjuvant and systemic therapies are urgently required. Endoplasmic reticulum (ER) stress is a potential therapeutic target, and two relatively new drugs, fenretinide and bortezomib (Velcade), each acting via different cellular mechanisms, induce ER stress leading to apoptosis in melanoma cells. The aim of this study was to test the hypothesis that apoptosis of melanoma cells may be increased by combining clinically achievable concentrations of fenretinide and bortezomib. Three human melanoma cell lines were used to assess changes in viability and the induction of apoptosis in response to fenretinide, bortezomib, or both drugs together. A s.c. xenograft model was used to test responses in vivo. Fenretinide and bortezomib synergistically decreased viability and increased apoptosis in all three melanoma lines at clinically achievable concentrations. This was also reflected by increased expression of GADD153, a marker of ER stress-induced apoptosis. In vivo, fenretinide in combination with bortezomib gave a marked reduction in xenograft tumor volume and an increase in apoptosis compared with fenretinide or bortezomib alone. The cell cycle stage of tumor cells in vivo were similar to that predicted from the effects of each drug or the combination in vitro. These results suggest that fenretinide and bortezomib, both of which are available in clinical formulation, warrant clinical evaluation as a combination therapy for metastatic melanoma.


Effects of cytarabine on activation of human T cells - cytarabine has concentration-dependent effects that are modulated both by valproic acid and all-trans retinoic acid.
Cytarabine is used in the treatment of acute myeloid leukemia (AML). Low-dose cytarabine can be combined with valproic acid and all-trans retinoic acid (ATRA) as AML-stabilizing treatment. We have investigated the possible risk of immunotoxicity by this combination. We examined the effects of cytarabine combined with valproic acid and ATRA on in vitro activated human T cells, and we tested cytarabine at concentrations reached during in vivo treatment with high doses, conventional doses and low doses. T cells derived from blood donors were activated in vitro in cell culture medium alone or supplemented with ATRA (1 μM), valproic acid (500 or 1000 μM) or cytarabine (0.01-44 μM). Cell characteristics were assessed by flow cytometry. Supernatants were analyzed for cytokines by ELISA or Luminex. Effects on primary human AML cell viability and proliferation of low-dose cytarabine (0.01-0.5 μM) were also assessed. Statistical tests include ANOVA and Cluster analyses. Only cytarabine 44 μM had both antiproliferative and proapoptotic effects. Additionally, this concentration increased the CD4:CD8 T cell ratio, prolonged the expression of the CD69 activation marker, inhibited CD95L and heat shock protein (HSP) 90 release, and decreased the release of several cytokines. In contrast, the lowest concentrations (0.35 and 0.01 μM) did not have or showed minor antiproliferative or cytotoxic effects, did not alter activation marker expression (CD38, CD69) or the release of CD95L and HSP90, but inhibited the release of certain T cell cytokines. Even when these lower cytarabine concentrations were combined with ATRA and/or valproic acid there was still no or minor effects on T cell viability. However, these combinations had strong antiproliferative effects, the expression of both CD38 and CD69 was altered and there was a stronger inhibition of the release of FasL, HSP90 as well as several cytokines. Cytarabine (0.01-0.05 μM) showed a dose-dependent antiproliferative effect on AML cells, and in contrast to the T cells this effect reached statistical significance even at 0.01 μM. Even low levels of cytarabine, and especially when combined with ATRA and valproic acid, can decrease T cell viability, alter activation-induced membrane-molecule expression and decrease the cytokine release.


Enzymatic transition states and transition state analog design.
All chemical transformations pass through an unstable structure called the transition state, which is poised between the chemical structures of the substrates and products. The transition states for chemical reactions are proposed to have lifetimes near 10(-13) sec, the time for a single bond vibration. No physical or spectroscopic method is available to directly observe the structure of the transition state for enzymatic reactions. Yet transition state structure is central to understanding catalysis, because enzymes function by lowering activation energy. An accepted view of enzymatic catalysis is tight binding to the unstable transition state structure. Transition state mimics bind tightly to enzymes by capturing a fraction of the binding energy for the transition state species. The identification of numerous transition state inhibitors supports the transition state stabilization hypothesis for enzymatic catalysis. Advances in methods for measuring and interpreting kinetic isotope effects and advances in computational chemistry have provided an experimental route to understand transition state structure. Systematic analysis of intrinsic kinetic isotope effects provides geometric and electronic structure for enzyme-bound transition states. This information has been used to compare transition states for chemical and enzymatic reactions; determine whether enzymatic activators alter transition state structure; design transition state inhibitors; and provide the basis for predicting the affinity of enzymatic inhibitors. Enzymatic transition states provide an understanding of catalysis and permit the design of transition state inhibitors. This article reviews transition state theory for enzymatic reactions. Selected examples of enzymatic transition states are compared to the respective transition state inhibitors.


Production of in vivo biotinylated scFv specific to almond (Prunus dulcis) proteins by recombinant Pichia pastoris.
The methylotropic yeast Pichia pastoris has demonstrated its suitability for large-scale production of recombinant proteins. As an eukaryotic organism P. pastoris presents a series of advantages at expression and processing of heterologous proteins when compared with Escherichia coli. In this work, P. pastoris has been used to express a scFv from a human synthetic library previously shown to bind almond proteins. In order to facilitate purification and post processing manipulations, the scFv was engineered with a C-terminal tag and biotinylated in vivo. After purification, biotinylated scFv were bound to avidin conjugated with HRP producing a multimeric scFv. The multimeric scFv showed to maintain their ability to recognize almond protein when assayed in ELISA, reaching a LOD of 470mgkg(-1). This study describes an easy method to produce large quantities of in vivo biotinylated scFv in P. pastoris. By substituting the enzyme or fluorochromes linked to avidin, it will be possible to generate a diverse number of multimeric scFv as probes to suit different analytical platforms in the detection of almond in food products.


Attempted isolation of a heparin proteoglycan from bovine liver capsule.
(1) Polysaccharides were isolated from bovine liver capsule by extraction with 2m-potassium chloride followed by precipitation from 0.8m-potassium chloride with cetylpyridinium chloride. Chondroitin sulphate was eliminated by digestion with hyaluronidase. The yield of heparin was approx. 40% of that obtained after extraction of the papain-digested tissue. (2) The macromolecular properties of the hyaluronidase-digested polysaccharide were studied by gel chromatography on Sephadex G-200 of the intact, as well as of the alkali-degraded, material. The results suggested the presence of single heparin chains in addition to a dermatan sulphate proteoglycan. (3) A purified heparin preparation was analysed for amino acids and neutral sugars. Xylose, galactose and serine were found in amounts corresponding to 0.1, 0.2, and 0.4 residue/polysaccharide chain (mol.wt. 7400), respectively. It is suggested that the isolated material had been degraded by a polysaccharidase with endo-enzyme properties.


[Sexually transmitted infections among transgender individuals and other sexual identities].
Few data are available regarding the prevalence of sexually transmitted infections (STI), including HIV-1 infection, and risk behaviors of transgender individuals. Previous reports indicate that this community has a high prevalence of HIV and STIs. Our objective was to compare the prevalence of HIV-1 infection, STI and risk behaviors of transgender people versus non transgender people. We used a cross sectional design study. Patients who received services at our testing site between November 2002 and April 2006, and provided written informed consent, were included in the analysis. Socio-demographic data, sexual behaviour, recreational drug use, condom use, concurrent or previous STI and HIV-1 infection diagnosis and partner serologic status, were collected. We used descriptive statistics and chi 2 for comparisons of proportions. In the period of the study, 105 transgender individuals were identified in a population of 4118 patients tested. The prevalence of HIV infection in the transgender group was 27.6% (29/105), while in the non transgender group was 6.2% (247/4013) p:0.0000. Low level of formal instruction, alcohol consumption, drug abuse, previous history of STI and sex work (100% transgenders and 2.3% of non-transgenders) were significantly more frequent in the transgender. The referred correct use of condom was similar in both groups. The prevalence of syphilis was 42.3% in transgender group and 18.1% in non-transgender individuals. These data show that this population has a very high prevalence of HIV-1 and STI. This information could be instrumental to design targets for intensive HIV prevention strategies in this particular high risk population.


Major cross-country differences in risk of dying for people with IDDM. Diabetes Epidemiology Research International Mortality Study Group.
Little is known concerning global differences in the risk of premature death for individuals developing youth-onset insulin-dependent diabetes mellitus (IDDM). The Diabetes Epidemiology Research International Study was developed to examine the mortality patterns of four population-based cohorts of IDDM cases from Allegheny County, Pennsylvania (n = 1000), Finland (n = 5146), Israel (n = 681), and Japan (n = 1428). All subjects were diagnosed as having diabetes, were less than 18 yr old at onset, were taking insulin at the time of hospital discharge, and were diagnosed between 1 January 1965 and 31 December 1979. The living status as of 1 January 1985 was determined. Overall, there were 182 deaths. Life-table analysis revealed that at 20-yr duration of diabetes, 5.5% of the cohort had died in Allegheny County in contrast to only 3.1% in Finland and 4.6% in Israel (P less than 0.01). Follow-up for an additional 3 yr in the United States and Finland revealed major differences in the 30- to 39-yr age-group, with 3.9 times greater premature mortality in the U.S. cohort compared with the Finnish group (overall mortality 2.3 vs. 0.6%, respectively). The Japanese cohort was developed in a somewhat different manner than the other three; therefore, the populations of the U.S., Finland, and Israel were reconfigured to make them directly comparable to that of Japan. The Japanese cohort exhibited markedly higher age-adjusted mortality rates (n/100,000 person-yr of diabetes) than the other three (Japan 681, U.S. 230, Finland 171, and Israel 131). These data indicate that young adult IDDM subjects are at an increased risk of premature death, there are differences in the mortality risk across countries, and both the U.S. and Japan have the major problem of an apparently excessive premature death rate among young people who have diabetes.


Effects of cosmetics containing purified honeybee (Apis mellifera L.) venom on acne vulgaris.
Acne vulgaris is a chronic dermatologic problem with multiple factors involved in its pathogenesis. Alternative solutions to acne treatment were instigated by antibiotic resistance despite of its extensive use. Purified bee venom (PBV) has been proposed as a promising candidate for that purpose. The present study was designed to confirm the antibacterial effect of PBV and access the efficacy of cosmetics containing PBV in subjects with acne vulgaris. The skin bacterium Propionibacterium acnes was incubated with PBV at various concentrations and bacterial growth was evaluated using the colony forming unit (CFU) assay. The mechanism of PBV employed in killing P. acnes was examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition, a total of 12 subjects were randomized in a double-blind, controlled trial to receive either cosmetics containing PBV or cosmetics without PBV for two weeks. Evaluations included lesion counts and skin microorganism. PBV exhibited antimicrobial activity in a concentration-dependent manner, reducing the number of P. acnes CFU by approximately 6 logs at a concentration of 0.5 mg. When PBV concentration was higher than 1.0 mg, no P. acnes colonies were spotted on an agar. TEM and SEM of untreated P. acnes illustrated the normal pleomorphic structure, whereas the PBV-treated bacterium lost the integrity of surface architecture. Significant difference (P=0.027) in the grading levels based on numbers of lesion counts for inflammatory and noninflammatory was observed in favour of the PBV group compared with the control group. In terms of average decrement of skin microorganism, subjects receiving cosmetics containing PBV experienced a significant 57.5% decrease of adenosine triphosphate levels, whereas participants receiving cosmetics without PBV experienced a nonsignificant decrease of 4.7%. These results show that the in vitro actions of antimicrobial activity of PBV were translated in vivo. Cosmetics containing PBV provided a certain degree of efficacy in terms of lesion counts and skin microorganism concentration compared with cosmetics without PBV in subjects with acne vulgaris. PBV may be a good candidate compound for developing therapeutic drug for the treatment of acne vulgaris.


Plasmids Shaped the Recent Emergence of the Major Nosocomial Pathogen Enterococcus faecium.
Enterococcus faecium is a gut commensal of humans and animals but is also listed on the WHO global priority list of multidrug-resistant pathogens. Many of its antibiotic resistance traits reside on plasmids and have the potential to be disseminated by horizontal gene transfer. Here, we present the first comprehensive population-wide analysis of the pan-plasmidome of a clinically important bacterium, by whole-genome sequence analysis of 1,644 isolates from hospital, commensal, and animal sources of E. faecium Long-read sequencing on a selection of isolates resulted in the completion of 305 plasmids that exhibited high levels of sequence modularity. We further investigated the entirety of all plasmids of each isolate (plasmidome) using a combination of short-read sequencing and machine-learning classifiers. Clustering of the plasmid sequences unraveled different E. faecium populations with a clear association with hospitalized patient isolates, suggesting different optimal configurations of plasmids in the hospital environment. The characterization of these populations allowed us to identify common mechanisms of plasmid stabilization such as toxin-antitoxin systems and genes exclusively present in particular plasmidome populations exemplified by copper resistance, phosphotransferase systems, or bacteriocin genes potentially involved in niche adaptation. Based on the distribution of k-mer distances between isolates, we concluded that plasmidomes rather than chromosomes are most informative for source specificity of E. faeciumIMPORTANCEEnterococcus faecium is one of the most frequent nosocomial pathogens of hospital-acquired infections. E. faecium has gained resistance against most commonly available antibiotics, most notably, against ampicillin, gentamicin, and vancomycin, which renders infections difficult to treat. Many antibiotic resistance traits, in particular, vancomycin resistance, can be encoded in autonomous and extrachromosomal elements called plasmids. These sequences can be disseminated to other isolates by horizontal gene transfer and confer novel mechanisms to source specificity. In our study, we elucidated the total plasmid content, referred to as the plasmidome, of 1,644 E. faecium isolates by using short- and long-read whole-genome technologies with the combination of a machine-learning classifier. This was fundamental to investigate the full collection of plasmid sequences present in our collection (pan-plasmidome) and to observe the potential transfer of plasmid sequences between E. faecium hosts. We observed that E. faecium isolates from hospitalized patients carried a larger number of plasmid sequences compared to that from other sources, and they elucidated different configurations of plasmidome populations in the hospital environment. We assessed the contribution of different genomic components and observed that plasmid sequences have the highest contribution to source specificity. Our study suggests that E. faecium plasmids are regulated by complex ecological constraints rather than physical interaction between hosts.


Effects of different protein content and glycaemic index of ad libitum diets on diabetes risk factors in overweight adults: the DIOGenes multicentre, randomized, dietary intervention trial.
Dietary regimens providing different levels of protein and glycemic index (GI) foods when prescribed for weight management may also influence insulin sensitivity. Overweight/obese adults in 8 European countries who lost ≥ 8% of initial body-weight (BW) after following a low calorie diet (LCD) were later randomly assigned with a 2x2 factorial design into 4 ad libitum dietary groups with two different protein content levels and dissimilar glycemic index, which were compared to a healthy reference diet. Specific markers assessing insulin resistance were measured. The LCD was initially applied to 932 adults and 773 were randomised to the 5 ad libitum diets. The 6-months programme was completed by 548 participants. The assignment to the Low Protein /High Glycemic Index diet induced a statistically higher HOMA-IR increase during the 6 months period as compared to the control. Contrariwise, the insulin response was lower in the High Protein/Low Glycemic Index diet after 60 and 90 min of an Oral Glucose Tolerance test subsequently carried out after the 6-months intervention. The Low Glycemic Index diets (either with high or low protein content) also lead to a decrease in fructosamine levels during the trial. After a weight loss period, an increase in the dietary protein proportions and a decrease in the consumption of foods with a high Glycemic Index within an ad libitum dietary intervention aiming to weight maintenance produced favorable effects on glycaemic control and insulin sensitivity in overweight/obese subjects.