Type 2 Innate Lymphoid Cells Impede IL-33-Mediated Tumor Suppression.
Although a number of studies have recently explored the contribution of the adaptive immunity in IL-33-mediated antitumor effects, innate immune involvement has been poorly characterized. Utilizing Rag1-/- mice (lacking T and B lymphocytes), we show in this study that either systemic administration of recombinant IL-33 or ectopic expression of IL-33 in melanoma cells is sufficient to inhibit tumor growth independent of adaptive antitumor immunity. We have demonstrated that IL-33-mediated antitumor effects depend on expansion and activation of NK cells. Interestingly, IL-33 also promoted the expansion of active type 2 innate lymphoid cells (ILC2s) via its receptor, ST2, which in turn inhibited NK activation and cytotoxicity. This IL-33-induced ILC2 activity coincided with greater expression of the immunosuppressive ectoenzyme CD73. Removal of CD73 from ILC2s in culture with NK cells resulted in markedly increased activation levels in NK cells, offering a potential mechanism by which ILC2s might suppress NK cell-mediated tumor killing. Thus, our data reveal an important contribution of IL-33-induced ILC2 to tumor growth by weakening NK cell activation and tumor killing, regardless of adaptive immunity.


Chemokine-Binding Proteins Encoded by Parapoxvirus of Red Deer of New Zealand Display Evidence of Gene Duplication and Divergence of Ligand Specificity.
Parapoxvirus of red deer in New Zealand (PVNZ) is a species of the Parapoxvirus genus that causes pustular dermatitis. We identified a cluster of genes in PVNZ that encode three unique chemokine-binding proteins (CBPs) namely ORF112.0, ORF112.3 and ORF112.6. Chemokines are a large family of molecules that direct cell trafficking to sites of inflammation and through lymphatic organs. The PVNZ-CBPs were analyzed by surface plasmon resonance against a broad spectrum of CXC, CC, XC and CX3C chemokines and were found to differ in their specificity and binding affinity. ORF112.0 interacted with chemokines from the CXC, CC and XC classes of chemokines with nM affinities. The ORF112.3 showed a preference for CXC chemokines, while ORF112.6 showed pM affinity binding for CC chemokines. Structural modeling analysis showed alterations in the chemokine binding sites of the CBPs, although the core structure containing two ß-sheets and three α-helices being conserved with the other parapoxvirus CBPs. Chemotaxis assays using neutrophils and monocytes revealed inhibitory impact of the CBPs on cell migration. Our results suggest that the PVNZ-CBPs are likely to have evolved through a process of gene duplication and divergence, and may have a role in suppressing inflammation and the anti-viral immune response.


Membrane attachment facilitates ligand access to the active site in monoamine oxidase A.
Monoamine oxidase membrane enzymes are responsible for the catalytic breakdown of extra- and intracellular neurotransmitters and are targets for the development of central nervous system drugs. We analyzed the dynamics of rat MAOA by performing multiple independent molecular dynamics simulations of membrane-bound and membrane-free forms to clarify the relationship between the mechanics of the enzyme and its function, with particular emphasis on the significance of membrane attachment. Principal component analysis of the simulation trajectories as well as correlations in the fluctuations of the residues pointed to the existence of three domains that define the global dynamics of the protein. Interdomain anticorrelated movements in the membrane-bound system facilitated the relaxation of interactions between residues surrounding the substrate cavity and induced conformational changes which expanded the active site cavity and opened putative pathways for substrate uptake and product release. Such events were less pronounced in the membrane-free system due to differences in the nature of the dominant modes of motion. The presence of the lipid environment is suggested to assist in decoupling the interdomain motions, consistent with the observed reduction in enzyme activity under membrane-free conditions. Our results are also in accordance with mutational analysis which shows that modifications of interdomain hinge residues decrease the activity of rat MAOA in solution.


Moments of speaking and silencing: Nurses share their experiences of manual handling in healthcare.
Nursing care involves complex patient handling tasks, resulting in high musculoskeletal injury rates. Epidemiological studies from the 1980s estimated a lifetime prevalence of lower back injuries for nurses between 35 and 80%. National and international studies continue to mirror these findings. Despite the development of programs intended to reduce manual handling injuries, sustainable solutions remain elusive. This paper reports on a study of nurses speaking about their perspectives on current manual handling practices. Qualitative research conducted in 2012 investigated nurses' perceptions and experiences relating to manual handling in the healthcare context and their participation in injury prevention programs. There were two research methods: semi-structured interviews and researcher reflective journaling. The research was framed in critical emancipatory methodology. Thirteen nurses from two Australian states participated in the study. Thematic analysis revealed an overarching theme of 'power relations' with a subcategory of '(mis)power' that comprised two subthemes, these being 'how to practice' and 'voicing practice issues'. Specifically, this paper explores nurses verbalising their views in the workplace and responses which left them feeling silenced, punished and disillusioned. The findings suggest that the sociopolitical context within which nurses practice impacts upon their ability to voice concerns or ideas related to manual handling. Inclusion of nurses in the manual handling dialogue may generate an expanded understanding of, and the potential to transform, manual handling practices in healthcare environments.


Tinnitus Retraining Therapy in Unilateral Tinnitus Patients with Single Side Deafness.
The purpose of this study was to evaluate the audiologic and clinical characteristics of the tinnitus in single-sided deafness (SSD) patients and to determine whether tinnitus retraining therapy (TRT) is beneficial to those patients who suffer from tinnitus with SSD. Thirty-six tinnitus patients with unilateral sensorineural profound hearing loss were included in the SSD group, and 46 tinnitus patients with normal hearing were included in the normal hearing group (NH group). Tinnitus evaluation consisted of the Tinnitus Handicap Inventory (THI), and visual analog scales (VAS) were obtained at the initial interview and 6 months after TRT. The therapeutic response was analyzed and compared between the two groups. The subjective intensity of tinnitus and objective loudness were greater in the SSD group. According to the THI scores prior to TRT, the SSD group showed significantly higher values, except in the emotional subscale. The THI score was significantly improved after TRT in both groups. The total THI score and all the subscale scores were significantly reduced. The VAS average scores were reduced from 6.5 to 5.1 in the SSD group, without statistical significance, whereas in the NH group, the VAS average scores significantly decreased from 5.8 to 4.5 after TRT. TRT was effective in SSD patients with subjective tinnitus.


Human prostate stem cell antigen and HSP70 fusion protein vaccine inhibits prostate stem cell antigen-expressing tumor growth in mice.
Prostate stem cell antigen (PSCA) has been considered a potentially worthwhile target for prostate cancer therapy with its overexpression in both androgen-dependent and androgen-independent prostate cancers. However, PSCA is an autoantigen that can evoke immunological tolerance and hardly incite effective immunologic response. In this study, we sought to construct the fusion protein vaccines based on PSCA and heat shock protein 70 (HSP70) and to evaluate their immune responses and therapeutic efficacy. A series of recombinant proteins were prepared, and then, the male C57BL/6 mice were immunized subcutaneously by inoculation with RM-PSCA/Luc cells. The PSCA-specific cellular immune responses were monitored with ELISPOT and intracellular cytokines staining assay, and ELISA assay was used to detect humoral immune responses. The tumor growth was observed by in vivo bioluminescence imaging. The results showed that the mice vaccinated with PSCA-HSP could induce the PSCA-specific cellular and humoral immune responses. Tumor progression could be quantitatively monitored by in vivo bioluminescence imaging. Animal experiments showed that PSCA-HSP could inhibit the growth of PSCA-expressing tumors and prolong the survival time of vaccinated mice. This study supported and confirmed the potential of HSP70 as a chaperone for protein vaccines, and PSCA-HSP could be of potential value for prostate cancer treatment.


A comparative analysis between laparoscopic and open adhesiolysis at a tertiary care center.
Laparotomy has been the favored approach regarding surgical management of small bowel obstruction (SBO); however, laparoscopy may offer improved outcomes. Patients undergoing laparoscopic lysis of adhesions (LOA) at our institution for SBO will have lower postoperative morbidity and 30-day mortality. Patients undergoing LOA at our institution, from 2000 to 2011, were reviewed. Categorical data were analyzed with χ(2) or Fisher's exact tests. Continuous data were analyzed with Student's t test or Wilcoxon rank sum. One hundred two (38 laparoscopic, 64 open) LOA cases were selected. Perioperative contamination and conservative management were higher in the open group. Open cases had a greater incidence of intensive care unit (ICU) admissions and longer length of stay. Stratified analysis determined a strong association between perioperative contamination and a higher incidence of ICU admission, perioperative contamination and longer LOS, and conservative management and longer LOS. Finally, patient outcome did not differ between those treated by surgeons trained in minimally invasive surgery (MIS) compared with those not trained in MIS. Careful consideration of surgical approach and timing is called for in all patients with SBO; however, whenever possible, laparoscopic preference should be given to most patients in an expeditious fashion irrespective of MIS training.


DNase I hypersensitive sites in the 5' flanking region of the human plasminogen activator inhibitor type 2 (PAI-2) gene are associated with basal and tumor necrosis factor-alpha-induced transcription in monocytes.
The plasminogen activator inhibitor type 2 (PAI-2) gene encodes a serine proteinase inhibitor (serpin) which is rapidly induced in response to the inflammatory cytokine, tumour necrosis factor-alpha (TNFalpha) in monocytes and macrophages. As an initial step towards understanding the molecular mechanisms underlying PAI-2 gene regulation in monocytes, we report here the analysis of the chromatin structure of 9.6 kb of 5' flanking region of the human PAI-2 gene for potential cis-acting regulatory regions using DNase I hypersensitivity mapping. Sites sensitive to DNase I were mapped in two monocytic cell lines representative of early monocytic differentiation; U937 cells, which synthesise low constitutive levels of PAI-2 that were induced following treatment with TNFalpha, and a MonoMac6 cell line which did not synthesise PAI-2 even after treatment with TNFalpha. Six DNase I hypersensitive sites (DHS) were identified; three upstream of the transcription initiation site (DH1, DH2, DH3) and three downstream of the transcription initiation site which were contained within intron A (DH4, DH5) and the exon 2/intron B junction (DH6). Among these, one distally located DH site (DH2) disappeared in both cell lines following treatment with TNFalpha. Two DH sites (DH1, DH6) were absent in PAI-2-producing U937 cells, but were present in MonoMac6 cells, which did not produce PAI-2, indicating the possible involvement of negative regulatory elements in the suppression of PAI-2 gene expression. The results demonstrate the involvement of chromatin structure in transcriptional responsiveness of the PAI-2 gene promoter and identify several loci which may be key control regions for PAI-2 gene transcription.


Helping psychiatry residents cope with patient suicide.
Every clinical specialty has its own high risk patient challenges that threaten to undermine their trainees' professional identity, evolving sense of competence. In psychiatric training, it is patient suicide, an all-too frequently encountered consequence of severe mental illness that may leave the treating resident perplexed, guilt-ridden, and uncertain of their suitability for the profession. This study evaluates a patient suicide training program aimed at educating residents about patient suicide, common reactions, and steps to attenuate emotional distress while facilitating learning. The intervention was selected aspects of a patient suicide educational program, "Collateral Damages,"-video vignettes, focused discussions, and a patient-based learning exercise. Pre- and post-survey results were compared to assess both knowledge and attitudes resulting from this educational program. Eight psychiatry residency training programs participated in the study, and 167 of a possible 240 trainees (response rate = 69.58 %) completed pre- and post-surveys. Knowledge of issues related to patient suicide increased after the program. Participants reported increased awareness of the common feelings physicians and trainees often experience after a patient suicide, of recommended "next" steps, available support systems, required documentation, and the role played by risk management. This patient suicide educational program increased awareness of issues related to patient suicide and shows promise as a useful and long overdue educational program in residency training. It will be useful to learn whether this program enhances patient care or coping with actual patient suicide. Similar programs might be useful for other specialties.


Human nuclease/helicase DNA2 alleviates replication stress by promoting DNA end resection.
In precancerous and cancerous lesions, excessive growth signals resulting from activation of oncogenes or loss of tumor suppressor genes lead to intensive replication stress, which is recognized by a high level of replication-associated DNA double-strand breaks (DSB). However, the molecular mechanism by which cells alleviate excessive replication stress remains unclear. In this study, we report that the human nuclease/helicase DNA2 facilitates homologous recombination to repair replication-associated DNA DSBs, thereby providing cells with survival advantages under conditions of replication stress. The nuclease activity of DNA2 was required for DSB end resection, which allowed subsequent recruitment of RPA and RAD51 to repair DSBs and restart replication. More importantly, DNA2 expression was significantly increased in human cancers and its expression correlated with patient outcome. Our findings therefore indicate that enhanced activity of DSB resection likely constitutes one mechanism whereby precancerous and cancerous cells might alleviate replication stress.


[Pharmacologic studies of antimotion sickness actions of ginger].
The pharmacologic actions related to antimotion sickness effects of ginger (Zingiber officinale Roscoe.) were studied. There was no significant effect on parameters of rotatory movement-induced electronystagmogram of rabbit after intravenous (i.v.) infection of ginger juice. The low amplitude fast wave pattern of electrocorticogram of rabbit changed to high amplitude slow wave pattern after i.v. injection of ginger juice. Rabbit gastric contraction in situ was shortly suppressed after ginger juice i.v. administration. In the isolated rat fundus strip preparations, however, ginger juice reduced the spontaneous contractile frequency, and enhanced the spontaneous contractile amplitude, which was followed by inhibition. Ginger juice produced longitudinal contraction of the guinea-pig isolated ileum, which was followed by rapid tachyphylaxis. This contraction effect was not affected by hexamethonium and 5-HT, but could be inhibited by cold storage, hyoscine, morphine, diphenhydramine, promethazine and substance P desensitization. Naloxone could eliminate this inhibition produced by morphine. By using dose-response relationship plot, non-competitive antagonisms were observed between ginger juice and Ach and between ginger juice and histamine in isolated guinea-pig ileum. It is suggested that the pungent constituents of ginger release substance P from sensory fibres. The released substance P in turn either stimulates cholinergic and histaminic neurons to release Ach and histamine, respectively, or produces direct muscle contraction by activating M and H1 receptors correspondingly. It is proposed that after being excited by substance P, M and H1 receptors are inactive temporarily and unable to be excited by agonists, therefore, ginger juice exhibits anticholinergic and antihistaminic action. Ginger juice produces antimotion sickness action possibly by central and peripheral anticholinergic and antihistaminic effects.


Differential-phase quantum key distribution experiment using a series of quantum entangled photon pairs.
We report what we believe to be the first differential-phase quantum key distribution experiment using a series of quantum entangled photon pairs. We employed two outstanding techniques. As an entangled photon source, we used a 1.5 microm band entangled photon pair source based on spontaneous four-wave mixing in a cooled dispersion-shifted fiber. As receivers, photon pairs were actively phase modulated with LiNbO3 phase modulators followed by very stable planar light-wave circuit Mach-Zehnder interferometers, which provided two nonorthogonal measurements. As a consequence, we successfully generated sifted keys with a quantum bit error rate of 8.3% and a key generation rate of 0.3 bit/s and revealed the feasibility of this QKD scheme.


Presidential leadership.
This chapter reviews psychological theories of leadership and selected literature on the American presidency to highlight key psychological principles of presidential leadership. Psychological theories, framed by the principles of leadership outlined by Freud (1921), include those of Burns (1978, 2003) on transformational leadership, Bass (1997) and House & Shamir (1993) on charismatic and transformational leadership, Gardner (1995) on stories of identity, Hogg (2001, 2003) on social identity, and Tyler & Lind (1992) on procedural justice. The discussion of presidential scholarship considers work by Barber (1992) on presidential character, Simonton (1986, 1987) on presidential personality and success, Skowronek (1997) on reconstructive politics, and Winter (1987) on presidential motive profiles. These studies suggest that followers have high expectations for presidents and that successful presidential leadership depends on opportunity, high levels of activity, intelligence, optimistic resilience, and flexibility.


Radiofrequency-induced carcinogenesis: cellular calcium homeostasis changes as a triggering factor.
The aim was to study the effects of radiofrequency (Rf) in a mice strain characterized by age-determined carcinogenesis of lymphatic tissues. Mice were treated with a 1?h/week Rf exposure for 4 months. A group submitted to sham exposure was used as control animals. The evolution of carcinogenesis was followed up to 18 months. The maximal life span of control mice was about 24 months. All dead animals were clinically and histologically examined to give an age-determined comparative quantification of the evolving carcinogenesis. A radiocalcium tracer method permitted the evaluation of Rf effects on transmembrane transport of extracellular calcium at 1 and 24 h after exposure. The determination of induced lipid peroxidation completed this second study. The findings show that Rf provoked an earlier general lymphocyte cell infiltration, formation of lymphoblastic ascites and extranodal tumours of different histological types, as well as an increased early mortality. The results suggest that in Rf-exposed mice, carcinogenesis may be induced earlier and with different pathological forms than in control animals. The modifications in cellular calcium homeostasis and the age-determined thymus involution appear to be important factors involved in this carcinogenesis process.


General application of the National Institute for Health and Clinical Excellence (NICE) guidance for active surveillance for men with prostate cancer is not appropriate in unscreened populations.
Study Type - Therapy (case series) Level of Evidence 4 What's known on the subject? and What does the study add? Active surveillance (AS) is a well-recognised management strategy to minimise the morbidity associated with radical treatment of prostate cancer. The National Institute for Health and Clinical Excellence guidelines initially suggested that all men with low-risk prostate cancer should first be offered AS. The cohort of men with upstaging and upgrading of prostate cancer from diagnosis to final pathology has been described in North American and European populations. As the rate of PSA testing in Britain is lower than North America and parts of Europe, the risk of more advanced disease at diagnosis of prostate cancer is higher. The present study is one of the first to examine this cohort in a British population and found the rate of features of advanced disease (extracapsular extension, seminal vesicle involvement and Gleason 4 + 3, or 8-10) to be 37.2%. To determine if the National Institute for Health and Clinical Excellence (NICE) guidelines for men with low-risk prostate cancer were generally applicable in unscreened populations. Retrospective analysis of prospectively collected case series from a single tertiary care centre in England. In all, 700 consecutive men treated for prostate cancer from 2005 by robot-assisted laparoscopic prostatectomy (RALP) were included. Patients satisfying NICE criteria for low-risk disease (PSA level < 10 ng/mL and Gleason score ≤ 6 and cT1-2a) had their pathological samples analysed for advanced disease, defined as extracapsular extension (ECE: pT3), seminal vesicle involvement (SVI), Gleason sum 7, or 8-10 or node-positive disease. In all, 275 patients (39.2%) met the NICE low-risk criteria, but pathologically advanced disease was found in 37.2% of this group. There was ECE in 71 patients (25.8%), 10 had SVI (3.6%), nine (3.3%) had Gleason score 7(4 + 3), and 12 had Gleason sum 8-10 (4.4%). The NICE guidance was developed largely on data from North America where populations are highly screened using PSA testing. In the UK, many men with low-risk disease features have high-risk disease and the general applicability of the NICE guidance is questionable in unscreened populations. We recommend that radical therapy is discussed as an alternative option to active surveillance.


Extracellular polymeric substances are transient media for microbial extracellular electron transfer.
Microorganisms exploit extracellular electron transfer (EET) in growth and information exchange with external environments or with other cells. Every microbial cell is surrounded by extracellular polymeric substances (EPS). Understanding the roles of three-dimensional (3D) EPS in EET is essential in microbiology and microbial exploitation for mineral bio-respiration, pollutant conversion, and bioenergy production. We have addressed these challenges by comparing pure and EPS-depleted samples of three representative electrochemically active strains viz Gram-negative Shewanella oneidensis MR-1, Gram-positive Bacillus sp. WS-XY1, and yeast Pichia stipites using technology from electrochemistry, spectroscopy, atomic force microscopy, and microbiology. Voltammetry discloses redox signals from cytochromes and flavins in intact MR-1 cells, whereas stronger signals from cytochromes and additional signals from both flavins and cytochromes are found after EPS depletion. Flow cytometry and fluorescence microscopy substantiated by N-acetylglucosamine and electron transport system activity data showed less than 1.5% cell damage after EPS extraction. The electrochemical differences between normal and EPS-depleted cells therefore originate from electrochemical species in cell walls and EPS. The 35 ± 15-nm MR-1 EPS layer is also electrochemically active itself, with cytochrome electron transfer rate constants of 0.026 and 0.056 s-1 for intact MR-1 and EPS-depleted cells, respectively. This surprisingly small rate difference suggests that molecular redox species at the core of EPS assist EET. The combination of all the data with electron transfer analysis suggests that electron "hopping" is the most likely molecular mechanism for electrochemical electron transfer through EPS.


Maternal and infant health problems after normal childbirth: a randomised controlled study in Zambia.
The main aim of the study was to discover if a midwife home visiting programme has a significant effect on the prevalence of health problems and breast feeding behaviour of mothers who delivered normally and their healthy fullterm newborn babies, during a period of 42 days after delivery. Another aim was to compare the mothers', the midwife's, and the doctor's findings of prevalence of health problems at the end of the puerperium period. A randomised controlled trial was carried out. One group of mothers and their infants were randomly allocated to a home visiting group (Group A); the other group (Group B) was only visited at day 42. The study was carried out at the University Teaching Hospital (UTH) in Lusaka, the capital city of Zambia. A total of 408 mothers who had a normal delivery and gave birth to a healthy fullterm infant, as assessed by the attending midwife, were randomised to two groups. Group A consisted of 208 mother/infant dyads who were visited by a midwife in their homes at days 3, 7, 28, and 42 after delivery and Group B consisted of 200 mother/infant dyads who were only visited at day 42. At day 42 an equal proportion (30%) of mothers in both groups perceived that they had health problems. The prevalence of infant health problems in Group B was significantly higher (p < 0.01) as perceived by mothers. There were more mothers in Group B (p < 0.01) perceiving insufficient milk production and giving supplementary feeding. At day 42, mothers in Group A (56%) took more actions than mothers in Group B (41%) to solve infant health problems (p < 0.03). In both groups the mothers' perceived own health problems, were significantly higher (p < 0.01) than those observed by the obstetrician and those observed by the midwife. The midwife found more infant health problems in Group B (p < 0.01) than in Group A and more infants with health problems in both groups compared with the paediatrician's findings (p < 0.01). There was a significant difference between the mothers' reported health problems and the health problems identified by the midwife and the doctors. The study shows that a midwife home visit and individual health education to mothers, reduce the prevalence of infant health problems, and enables the mother to more often take action when an infant health problem is identified. There is a need to re-evaluate the midwifery training curriculums with the intention to include more infant management care.


Rapid Adaptation and Remote Delivery of Undergraduate Research Training during the COVID-19 Pandemic.
When COVID-19 caused worldwide cancellations of summer research immersion programs in 2020, Mayo Clinic rallied to create an alternate virtual experience called Summer Foundations in Research (SFIR). SFIR was designed not only to ensure the continuance of science pathways training for undergraduate scientists but also to support undergraduate mental wellbeing, given the known pandemic stressors. A total of 170 participants took part in the program and were surveyed pre-post for outcomes in biomedical research career knowledge, biomedical research career interest, research skills confidence, and three dimensions of mental wellbeing. Knowledge of and interest in careers involving biomedical research rose significantly following participation in SFIR. The participants' mean research skills confidence also rose between 0.08 and 1.32 points on a 7-point scale across 12 items from the Clinical Research Appraisal Inventory. Success in science pathways support was accompanied by positive shifts in participant mental wellbeing. Measurable decreases in stress (Perceived Stress Scale, p < 0.0001) accompanied gains in resilience (Brief Resilience Scale, p < 0.0001) and life satisfaction (Satisfaction with Life Scale, p = 0.0005). Collectively, the data suggest that core objectives of traditional in-person summer research programming can be accomplished virtually and that these programs can simultaneously impact student wellbeing. This theoretical framework is particularly salient during COVID-19, but the increased accessibility of virtual programs such as SFIR can continue to bolster science education pathways long after the pandemic is gone.


Deep learning approach to classification of lung cytological images: Two-step training using actual and synthesized images by progressive growing of generative adversarial networks.
Cytology is the first pathological examination performed in the diagnosis of lung cancer. In our previous study, we introduced a deep convolutional neural network (DCNN) to automatically classify cytological images as images with benign or malignant features and achieved an accuracy of 81.0%. To further improve the DCNN's performance, it is necessary to train the network using more images. However, it is difficult to acquire cell images which contain a various cytological features with the use of many manual operations with a microscope. Therefore, in this study, we aim to improve the classification accuracy of a DCNN with the use of actual and synthesized cytological images with a generative adversarial network (GAN). Based on the proposed method, patch images were obtained from a microscopy image. Accordingly, these generated many additional similar images using a GAN. In this study, we introduce progressive growing of GANs (PGGAN), which enables the generation of high-resolution images. The use of these images allowed us to pretrain a DCNN. The DCNN was then fine-tuned using actual patch images. To confirm the effectiveness of the proposed method, we first evaluated the quality of the images which were generated by PGGAN and by a conventional deep convolutional GAN. We then evaluated the classification performance of benign and malignant cells, and confirmed that the generated images had characteristics similar to those of the actual images. Accordingly, we determined that the overall classification accuracy of lung cells was 85.3% which was improved by approximately 4.3% compared to a previously conducted study without pretraining using GAN-generated images. Based on these results, we confirmed that our proposed method will be effective for the classification of cytological images in cases at which only limited data are acquired.


Mechanism of RNA polymerase II bypass of oxidative cyclopurine DNA lesions.
In human cells, the oxidative DNA lesion 8,5'-cyclo-2'-deoxyadenosine (CydA) induces prolonged stalling of RNA polymerase II (Pol II) followed by transcriptional bypass, generating both error-free and mutant transcripts with AMP misincorporated immediately downstream from the lesion. Here, we present biochemical and crystallographic evidence for the mechanism of CydA recognition. Pol II stalling results from impaired loading of the template base (5') next to CydA into the active site, leading to preferential AMP misincorporation. Such predominant AMP insertion, which also occurs at an abasic site, is unaffected by the identity of the 5'-templating base, indicating that it derives from nontemplated synthesis according to an A rule known for DNA polymerases and recently identified for Pol II bypass of pyrimidine dimers. Subsequent to AMP misincorporation, Pol II encounters a major translocation block that is slowly overcome. Thus, the translocation block combined with the poor extension of the dA.rA mispair reduce transcriptional mutagenesis. Moreover, increasing the active-site flexibility by mutation in the trigger loop, which increases the ability of Pol II to accommodate the bulky lesion, and addition of transacting factor TFIIF facilitate CydA bypass. Thus, blocking lesion entry to the active site, translesion A rule synthesis, and translocation block are common features of transcription across different bulky DNA lesions.


Cranberry (Vaccinium macrocarpon) extract treatment improves triglyceridemia, liver cholesterol, liver steatosis, oxidative damage and corticosteronemia in rats rendered obese by high fat diet.
Obese individuals have higher production of reactive oxygen species, which leads to oxidative damage. We hypothesize that cranberry extract (CE) can improve this dysfunction in HFD-induced obesity in rats since it has an important antioxidant activity. Here, we evaluated the effects of CE in food intake, adiposity, biochemical and hormonal parameters, lipogenic and adipogenic factors, hepatic morphology and oxidative balance in a HFD model. At postnatal day 120 (PN120), male Wistar rats were assigned into two groups: (1) SD (n = 36) fed with a standard diet and (2) HFD (n = 36), fed with a diet containing 44.5% (35.2% from lard) energy from fat. At PN150, 12 animals from SD and HFD groups were killed while the others were subdivided into four groups (n = 12/group): animals that received 200 mg/kg cranberry extract (SD CE, HFD CE) gavage/daily/30 days or water (SD, HFD). At PN180, animals were killed. HFD group showed higher body mass and visceral fat, hypercorticosteronemia, higher liver glucocorticoid sensitivity, cholesterol and triglyceride contents and microsteatosis. Also, HFD group had higher lipid peroxidation (plasma and tissues) and higher protein carbonylation (liver and adipose tissue) compared to SD group. HFD CE group showed lower body mass gain, hypotrygliceridemia, hypocorticosteronemia, and lower hepatic cholesterol and fatty acid synthase contents. HFD CE group displayed lower lipid peroxidation, protein carbonylation (liver and adipose tissue) and accumulation of liver fat compared to HFD group. Although adiposity was not completely reversed, cranberry extract improved the metabolic profile and reduced oxidative damage and steatosis in HFD-fed rats, which suggests that it can help manage obesity-related disorders.


Serotypes and virulence gene profiles of shiga toxin-producing Escherichia coli strains isolated from feces of pasture-fed and lot-fed sheep.
Shiga toxin-producing Escherichia coli (STEC) strains possessing genes for enterohemolysin (ehxA) and/or intimin (eae), referred to here as complex STEC (cSTEC), are more commonly recovered from the feces of humans with hemolytic uremic syndrome and hemorrhagic colitis than STEC strains that do not possess these accessory virulence genes. Ruminants, particularly cattle and sheep, are recognized reservoirs of STEC populations that may contaminate foods destined for human consumption. We isolated cSTEC strains from the feces of longitudinally sampled pasture-fed sheep, lot-fed sheep maintained on diets comprising various combinations of silage and grain, and sheep simultaneously grazing pastures with cattle to explore the diversity of cSTEC serotypes capable of colonizing healthy sheep. A total of 67 cSTEC serotypes were isolated, of which 21 (31.3%), mainly isolated from lambs, have not been reported. Of the total isolations, 58 (86.6%) were different from cSTEC serotypes isolated from a recent study of longitudinally sampled healthy Australian cattle (M. Hornitzky, B. A. Vanselow, K. Walker, K. A. Bettelheim, B. Corney, P. Gill, G. Bailey, and S. P. Djordjevic, Appl. Environ. Microbiol. 68:6439-6445, 2002). Our data suggest that cSTEC serotypes O5:H(-), O75:H8, O91:H(-), O123:H(-), and O128:H2 are well adapted to colonizing the ovine gastrointestinal tract, since they were the most prevalent serotypes isolated from both pasture-fed and lot-fed sheep. Collectively, our data show that Australian sheep are colonized by diverse cSTEC serotypes that are rarely isolated from healthy Australian cattle.


Antenatal lifestyle advice for women who are overweight or obese: LIMIT randomised trial.
To determine the effect of antenatal dietary and lifestyle interventions on health outcomes in overweight and obese pregnant women. Multicentre randomised trial. We utilised a central telephone randomisation server, with computer generated schedule, balanced variable blocks, and stratification for parity, body mass index (BMI) category, and hospital. Three public maternity hospitals across South Australia. 2212 women with a singleton pregnancy, between 10+0 and 20+0 weeks' gestation, and BMI ≥ 25. 1108 women were randomised to a comprehensive dietary and lifestyle intervention delivered by research staff; 1104 were randomised to standard care and received pregnancy care according to local guidelines, which did not include such information. Incidence of infants born large for gestational age (birth weight ≥ 90th centile for gestation and sex). Prespecified secondary outcomes included birth weight >4000 g, hypertension, pre-eclampsia, and gestational diabetes. Analyses used intention to treat principles. 2152 women and 2142 liveborn infants were included in the analyses. The risk of the infant being large for gestational age was not significantly different in the two groups (lifestyle advice 203/1075 (19%) v standard care 224/1067 (21%); adjusted relative risk 0.90, 95% confidence interval 0.77 to 1.07; P=0.24). Infants born to women after lifestyle advice were significantly less likely to have birth weight above 4000 g (lifestyle advice 164/1075 (15%) v standard care 201/1067 (19%); 0.82, 0.68 to 0.99; number needed to treat (NNT) 28, 15 to 263; P=0.04). There were no differences in maternal pregnancy and birth outcomes between the two treatment groups. For women who were overweight or obese, the antenatal lifestyle advice used in this study did not reduce the risk delivering a baby weighing above the 90th centile for gestational age and sex or improve maternal pregnancy and birth outcomes. Australian and New Zealand Clinical Trials Registry (ACTRN12607000161426).


Targeting antisense mitochondrial noncoding RNAs induces bladder cancer cell death and inhibition of tumor growth through reduction of survival and invasion factors.
Knockdown of the antisense noncoding mitochondrial RNAs (ASncmtRNAs) induces apoptotic death of several human tumor cell lines, but not normal cells, supporting a selective therapy against different types of cancer. In this work, we evaluated the effects of knockdown of ASncmtRNAs on bladder cancer (BCa). We transfected the BCa cell lines UMUC-3, RT4 and T24 with the specific antisense oligonucleotide Andes-1537S, targeted to the human ASncmtRNAs. Knockdown induced a strong inhibition of cell proliferation and increase in cell death in all three cell lines. As observed in UMUC-3 cells, the treatment triggered apoptosis, evidenced by loss of mitochondrial membrane potential and Annexin V staining, along with activation of procaspase-3 and downregulation of the anti-apoptotic factors survivin and Bcl-xL. Treatment also inhibited cell invasion and spheroid formation together with inhibition of N-cadherin and MMP 11. In vivo treatment of subcutaneous xenograft UMUC-3 tumors in NOD/SCID mice with Andes-1537S induced inhibition of tumor growth as compared to saline control. Similarly, treatment of a high-grade bladder cancer PDX with Andes-1537S resulted in a strong inhibition of tumor growth. Our results suggest that ASncmtRNAs could be potent targets for bladder cancer as adjuvant therapy.


Inhibition of agrin-induced acetylcholine-receptor aggregation by heparin, heparan sulfate, and other polyanions.
Heparin and heparan sulfate have been shown to block nerve-induced acetylcholine-receptor (AChR) aggregation at developing neuromuscular junctions. We found that heparin, heparan sulfate, and a wide variety of other polyanions also inhibited agrin-induced AChR aggregation. The more highly charged the polyanion, the more potent it was as an inhibitor. Inhibition of agrin-induced AChR aggregation was due, at least in part, to the formation of a complex between the polyanion and agrin that was inactive. These findings are consistent with the hypothesis that nerve-induced aggregation of AChRs is mediated by the release of agrin, or a closely related protein, from axon terminals and suggest that a polyanion, such as a sulfated proteoglycan, may be involved in the interaction of agrin with its receptor on the myotube surface.


Tyrosine-based signal mediates LRP6 receptor endocytosis and desensitization of Wnt/β-catenin pathway signaling.
Wnt/β-catenin signaling orchestrates a number of critical events including cell growth, differentiation, and cell survival during development. Misregulation of this pathway leads to various human diseases, specifically cancers. Endocytosis and phosphorylation of the LDL receptor-related protein 6 (LRP6), an essential co-receptor for Wnt/β-catenin signaling, play a vital role in mediating Wnt/β-catenin signal transduction. However, its regulatory mechanism is not fully understood. In this study, we define the mechanisms by which LRP6 endocytic trafficking regulates Wnt/β-catenin signaling activation. We show that LRP6 mutant with defective tyrosine-based signal in its cytoplasmic tail has an increased cell surface distribution and decreased endocytosis rate. These changes in LRP6 endocytosis coincide with an increased distribution to caveolae, increased phosphorylation, and enhanced Wnt/β-catenin signaling. We further demonstrate that treatment of Wnt3a ligands or blocking the clathrin-mediated endocytosis of LRP6 leads to a redistribution of wild-type receptor to lipid rafts. The LRP6 tyrosine mutant also exhibited an increase in signaling activation in response to Wnt3a stimulation when compared with wild-type LRP6, and this activation is suppressed when caveolae-mediated endocytosis is blocked. Our results reveal molecular mechanisms by which LRP6 endocytosis routes regulate its phosphorylation and the strength of Wnt/β-catenin signaling, and have implications on how this pathway can be modulated in human diseases.


A MicroRNA302-367-Erk1/2-Klf2-S1pr1 Pathway Prevents Tumor Growth via Restricting Angiogenesis and Improving Vascular Stability.
Angiogenic hypersprouting and leaky vessels are essential for tumor growth. MicroRNAs have unique therapeutic advantages by targeting multiple pathways of tumor-associated angiogenesis, but the function of individual miRNAs of miR302-367 cluster in angiogenesis and tumors has not yet been fully evaluated. To investigate the functions of miR302-367 in developmental angiogenesis and tumor angiogenesis and explore the molecular mechanisms of microRNA for the treatment of pathological neovascularization-related diseases. Here, we show that miR302-367 elevation in endothelial cells reduces retinal sprouting angiogenesis and promotes vascular stability in vivo, ex vivo, and in vitro. Erk1/2 is identified as direct target of miR302-367, and downregulation of Erk1/2 on miR302-367 elevation in endothelial cells increases the expression of Klf2 and in turn S1pr1 and its downstream target VE-cadherin, suppressing angiogenesis and improving vascular stability. Conversely, both pharmacological blockade and genetic deletion of S1pr1 in endothelial cells reverse the antiangiogenic and vascular stabilizing effect of miR302-367 in mice. Tumor angiogenesis shares features of developmental angiogenesis, and endothelial specific elevation of miR302-367 reduces tumor growth by restricting sprout angiogenesis and decreasing vascular permeability via the same Erk1/2-Klf2-S1pr1 pathways. MiR302-367 regulation of an Erk1/2-Klf2-S1pr1 pathway in the endothelium advances our understanding of angiogenesis, meanwhile also provides opportunities for therapeutic intervention of tumor growth.


Restrictions upon Epstein-Barr virus infection of the leukemic cell are demonstrated in patients with hairy cell leukemia.
We tested the hypothesis that Epstein-Barr virus (EBV) might actually infect leukemic hairy cells in vivo by examining those cells for the EBV-receptor, EBV nuclear antigen (EBNA) and membrane antigen (MA), for spontaneous transformation and rescue of infectious virus and for presence of EBV genome. EBV-receptors were found on subpopulations of leukemic cells from each of 7 patients with hairy cell leukemia (HCL) tested. MA was present on low numbers (1-5 per cent) of fresh leukemic cells of 7 patients and in some instances occurred with a greater frequency after 3 to 5 days in culture, with or without 12-O-tetradecanoylphorbol-13-acetate. In 11 fresh leukemic cell preparations from 8 HCL patients, no EBNA was demonstrated. However, 2 samples after 4 days in culture expressed low frequencies of EBNA-positive cells. Spontaneous, EBV-positive cell lines were established with a high transformation efficiency from 3 HCL blood samples but not from 8 other specimens. Infectious EBV could be rescued from some hairy leukemic cell preparations by co-cultivation with cord blood lymphocytes. These results demonstrated that leukemic cell populations harbored infectious EBV, that the leukemic cells expressed virus receptors and suggested that a small subpopulation of leukemic cells might become infected in vivo at least transiently and possibly transformed in vitro by EBV. To test for the extent of occult in vivo infection of leukemic cells with EBV, Southern type hybridization studies were performed with a probe for EBV genome (Bam HI W). At a sensitivity level of 0.1 genome per cell, EBV genome was not detected in the leukemic cell populations of 7 patients. We conclude that host defence mechanisms protecting these individuals from EBV also prevent infections of the leukemic cell and/or most hairy leukemic cells are not suitable targets for both infection and transformation.


Nano-assembly of amyloid β peptide: role of the hairpin fold.
Structural investigations have revealed that β hairpin structures are common features in amyloid fibrils, suggesting that these motifs play an important role in amyloid assembly. To test this hypothesis, we characterized the effect of the hairpin fold on the aggregation process using a model β hairpin structure, consisting of two Aβ(14-23) monomers connected by a turn forming YNGK peptide. AFM studies of the assembled aggregates revealed that the hairpin forms spherical structures whereas linear Aβ(14-23) monomers form fibrils. Additionally, an equimolar mixture of the monomer and the hairpin assembles into non-fibrillar aggregates, demonstrating that the hairpin fold dramatically changes the morphology of assembled amyloid aggregates. To understand the molecular mechanism underlying the role of the hairpin fold on amyloid assembly, we performed single-molecule probing experiments to measure interactions between hairpin and monomer and two hairpin complexes. The studies reveal that the stability of hairpin-monomer complexes is much higher than hairpin-hairpin complexes. Molecular dynamics simulations revealed a novel intercalated complex for the hairpin and monomer and Monte Carlo modeling further demonstrated that such nano-assemblies have elevated stability compared with stability of the dimer formed by Aβ(14-23) hairpin. The role of such folding on the amyloid assembly is also discussed.


Managing chronic pathologies with a stepped mHealth-based approach in clinical psychology and medicine.
Chronic diseases and conditions typically require long-term monitoring and treatment protocols both in traditional settings and in out-patient frameworks. The economic burden of chronic conditions is a key challenge and new and mobile technologies could offer good solutions. mHealth could be considered an evolution of eHealth and could be defined as the practice of medicine and public health supported by mobile communication devices. mHealth approach could overcome limitations linked with the traditional, restricted, and highly expensive in-patient treatment of many chronic pathologies. Possible applications include stepped mHealth approach, where patients can be monitored and treated in their everyday contexts. Unfortunately, many barriers for the spread of mHealth are still present. Due the significant impact of psychosocial factors on disease evolution, psychotherapies have to be included into the chronic disease protocols. Existing psychological theories of health behavior change have to be adapted to the new technological contexts and requirements. In conclusion, clinical psychology and medicine have to face the "chronic care management" challenge in both traditional and mHealth settings.


Fluorinated Cerium(IV) Enaminolates: Alternative Precursors for Chemical Vapor Deposition of CeO2 Thin Films.
High-yield synthesis of four new fluorinated enaminones LH2 (RfC(O)C2H2NH)2C2H4 (Rf = CF3 (2a), C2F5 (2b), C3F7 (2c)) and (F3CC(O)C2H2NH)2C3H6 (2a') as dianionic ligands is described. The ligands were characterized in solution (via nuclear magnetic resoannce (NMR)) as well as in the solid state (via X-ray diffraction (XRD)). The ligating ability of the enaminones was verified by reacting them with [Ce2(O(i)Pr)8(HO(i)Pr)2], which resulted in monomeric cerium(IV) complexes [CeL2] (3a-c, 3a') based on tetradentate chelation of the ligands. Cerium enaminolates were comprehensively analyzed by NMR spectroscopy, mass spectrometry, and single-crystal XRD studies to verify their monomeric nature. High stability under ambient conditions and high volatility makes them a potential precursor for the gas-phase synthesis of CeO2. Complexes 3a and 3b were applied as precursors in thermal and plasma-enhanced chemical vapor deposition to obtain crystalline ceria films with different surface morphologies. The purity and surface states of the films were analyzed by X-ray photoelectron spectroscopy, which revealed a high amount of Ce(3+) on the subsurface of CeO2 films.


What is a Good Looking Penis? How Women Rate the Penile Appearance of Men with Surgically Corrected Hypospadias.
Some men with corrected hypospadias perceive their penile appearance to be abnormal, although health professionals consider these results satisfactory. The aim of this study was to investigate how relevant women consider single aspects of penile appearance to be. Moreover, we studied whether women perceive hypospadias-affected surgically repaired genitals (HASRGs) to be as normal-looking as circumcised genitals and identified the most relevant predictors that influence whether a penis is perceived as normal. In this cross-sectional study, 105 women in different age groups (age range: 16-20, 25-30, and 40-45 years) completed a standardized questionnaire. Participants had to rate the importance of eight penile aspects and to indicate how normal they found the appearance of standardized photos of 10 HASRGs and of 10 circumcised genitals. Furthermore, they were asked about demographic characteristics and their sexuality. Results showed that women considered the position and shape of the meatus as the least important penile aspect. Furthermore, results showed that HASRGs with distal hypospadias were perceived to be as normal-looking as circumcised genitals, whereas genitals with more proximal hypospadias were perceived as significantly less normal. However, the difference need not be considered clinically relevant, as the effect size was small. Observer-related predictors of a more positive perception of HASRGs were higher age, higher sexual interest, and perceiving the general cosmetic penile appearance as more important and penile length as less important. Overall, women were found to consider the "position and shape of the meatus" as the least important penile aspect. These findings may stimulate reflections regarding the relevance of surgical correction of the meatus in minor forms of hypospadias. In addition, this study indicates that women perceived genitals of men with distal operated hypospadias (which represents the majority of hypospadias) to be as normal as nonaffected, circumcised genitals.


Experimental anterior uveitis after subcutaneous injection of feline sarcoma virus.
Feline sarcoma virus (FeSV) is a naturally occurring virus that causes spontaneous tumors in cats. The immunologic and morphologic characteristics of these tumors have been studied extensively. It was recently observed in experiments undertaken to induce systemic malignancy with this virus, that severe uveitis and clinical blindness occurred. An investigation of the ophthalmologic changes was undertaken. A fulminent anterior uveitis was produced in cats by a series of subcutaneous injections of live FeLV-FeSV. This intraocular inflammation occurred in five of six animals using high viral titers, and four of seven with lower titers, resulting from the freeze thaw process. On histopathologic examination, most animals demonstrated dysplastic changes of the ciliary body in addition to the iridocyclitis. The remainder of the eye was unaffected. These animals developed systemic tumors unaccompanied by local inflammation, many of which spontaneously regressed. Notable features of this potential model for uveitis are that (1) direct injection into the eye is unnecessary, and (2) intravenous administration inducing immune tolerance with antigenic overload presented to the spleen is avoided. This inflammatory reaction seems to be specific to the iris and ciliary body. Levels of live virus detected in the aqueous humor exceeded those in the serum. These results suggest that the virus may be actively secreted by the ciliary epithelium, or may preferentially proliferate within the eye.


Metronomic gemcitabine in combination with sunitinib inhibits multisite metastasis and increases survival in an orthotopic model of pancreatic cancer.
Metronomic chemotherapy suppresses growth of primary tumors and established metastases. However, its effect on metastatic progression is essentially unknown. We report the treatment of a metastatically competent model of pancreatic cancer with metronomic gemcitabine and sunitinib. Mice with orthotopic, red fluorescent protein-expressing, pancreatic cancer tumorgrafts were treated with gemcitabine on a metronomic (1 mg/kg daily, METG) or maximum tolerated dose (150 mg/kg twice weekly, MTDG) schedule with or without sunitinib (SU). Rates of primary tumor growth, metastasis, ascites, and survival were calculated. Gemcitabine at a daily dose of 2 mg or greater led to toxicity within 1 month in mice without tumors but METG at 1 mg/kg/d was well tolerated. Mice with pancreatic cancer tumorgrafts died with metastatic disease at a median of 25 days. METG/SU significantly prolonged median overall survival (44 days) compared with control or either regimen alone (P < 0.05). Primary tumor growth was inhibited by METG/SU (P = 0.03) but neither METG nor sunitinib alone. In contrast, treatment with METG suppressed metastasis at multiple sites, an effect enhanced by sunitinib. MTDG with or without sunitinib had the most favorable effect on primary tumor growth and survival, but its antimetastatic efficacy was similar to that of METG/SU. von Willebrand factor expression was inhibited by METG. Antimetastatic activity approaching that of MTDG is achieved with a total dose reduced 42 times using METG and is further enhanced by sunitinib. Our results suggest the potential of this therapeutic paradigm against pancreatic cancer in the adjuvant and maintenance settings.


Secretion of Plasmodium falciparum rhoptry protein into the plasma membrane of host erythrocytes.
The rhoptry is an organelle of the malarial merozoite which has been suggested to play a role in parasite invasion of its host cell, the erythrocyte. A monoclonal antibody selected for reactivity with this organelle identifies a parasite synthesized protein of 110 kD. From biosynthetic labeling experiments it was demonstrated that the protein is synthesized midway through the erythrocytic cycle (the trophozoite stage) but immunofluorescence indicates the protein is not localized in the organelle until the final stage (segmenter stage) of intraerythrocytic development. Immunoelectron microscopy shows that the protein is localized in the matrix of the rhoptry organelle and on membranous whorls secreted from the merozoite. mAb recognition of the protein is dithiothreitol (DTT) labile, indicating that the conformation of the epitope is dependent on a disulfide linkage. During erythrocyte reinvasion by the extracellular merozoite, immunofluorescence shows the rhoptry protein discharging from the merozoite and spreading around the surface of the erythrocyte. The protein is located in the plasma membrane of the newly invaded erythrocyte. These studies suggest that the 110-kD rhoptry protein is inserted into the membrane of the host erythrocyte during merozoite invasion.


Invasive aspergillosis in patients admitted to the intensive care unit with severe influenza: a retrospective cohort study.
Invasive pulmonary aspergillosis typically occurs in an immunocompromised host. For almost a century, influenza has been known to set up for bacterial superinfections, but recently patients with severe influenza were also reported to develop invasive pulmonary aspergillosis. We aimed to measure the incidence of invasive pulmonary aspergillosis over several seasons in patients with influenza pneumonia in the intensive care unit (ICU) and to assess whether influenza was an independent risk factor for invasive pulmonary aspergillosis. We did a retrospective multicentre cohort study. Data were collected from adult patients with severe influenza admitted to seven ICUs across Belgium and The Netherlands during seven influenza seasons. Patients were older than 18 years, were admitted to the ICU for more than 24 h with acute respiratory failure, had pulmonary infiltrates on imaging, and a confirmed influenza infection based on a positive airway PCR test (influenza cohort). We used logistic regression analyses to determine if influenza was independently associated with invasive pulmonary aspergillosis in non-immunocompromised (ie, no European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group [EORTC/MSG] host factor) influenza-positive patients (influenza case group) compared with non-immunocompromised patients with severe community-acquired pneumonia who had a negative airway influenza PCR test (control group). Data were collected from patients admitted to the ICU between Jan 1, 2009, and June 30, 2016. Invasive pulmonary aspergillosis was diagnosed in 83 (19%) of 432 patients admitted with influenza (influenza cohort), a median of 3 days after admission to the ICU. The incidence was similar for influenza A and B. For patients with influenza who were immunocompromised, incidence of invasive pulmonary aspergillosis was as high as 32% (38 of 117 patients), whereas in the non-immunocompromised influenza case group, incidence was 14% (45 of 315 patients). Conversely, only 16 (5%) of 315 patients in the control group developed invasive pulmonary aspergillosis. The 90-day mortality was 51% in patients in the influenza cohort with invasive pulmonary aspergillosis and 28% in the influenza cohort without invasive pulmonary aspergillosis (p=0·0001). In this study, influenza was found to be independently associated with invasive pulmonary aspergillosis (adjusted odds ratio 5·19; 95% CI 2·63-10·26; p<0·0001), along with a higher APACHE II score, male sex, and use of corticosteroids. Influenza was identified as an independent risk factor for invasive pulmonary aspergillosis and is associated with high mortality. Future studies should assess whether a faster diagnosis or antifungal prophylaxis could improve the outcome of influenza-associated aspergillosis. None.


Application of fluoroquinolone pharmacodynamics.
Pharmacodynamics provides a rational basis for optimizing dosing regimens by describing the relationship between drug, host and antimicrobial effect. The successful identification of meaningful pharmacodynamic outcome parameters can, therefore, greatly assist clinicians in making objective prescribing decisions rather than relying on static in vitro MIC data. While pharmacodynamic outcome parameters have been proposed for select antimicrobial agents, their clinical application remains to be defined fully. Quinolone antibiotics are generally considered to have concentration-dependent bactericidal activity and peak/MIC and AUC/MIC ratios have been identified as possible pharmacodynamic predictors of clinical and microbiological outcome as well as the development of bacterial resistance. Investigators have suggested that AUC/MIC ratios of 100-125 or peak/MIC ratios of >10 are required to predict clinical and microbiological success and to limit the development of bacterial resistance. These conclusions are derived primarily from studies of Gram-negative bacteria, and recent data suggest that these ratios may not be applicable for Streptococcus pneumoniae, where an AUC/MIC ratio of <40 appears to be a more accurate predictor. There is considerable variation in pharmacodynamic calculations and outcome parameters appear to be quinolone- and pathogen-specific. Additional prospective clinical research is needed to characterize quinolone pharmacodynamic parameters and answer unresolved questions regarding optimal pharmacodynamic outcome predictors for Gram-positive bacteria, anaerobes and atypical respiratory pathogens.


Decreased Brain Levels of Vitamin B12 in Aging, Autism and Schizophrenia.
Many studies indicate a crucial role for the vitamin B12 and folate-dependent enzyme methionine synthase (MS) in brain development and function, but vitamin B12 status in the brain across the lifespan has not been previously investigated. Vitamin B12 (cobalamin, Cbl) exists in multiple forms, including methylcobalamin (MeCbl) and adenosylcobalamin (AdoCbl), serving as cofactors for MS and methylmalonylCoA mutase, respectively. We measured levels of five Cbl species in postmortem human frontal cortex of 43 control subjects, from 19 weeks of fetal development through 80 years of age, and 12 autistic and 9 schizophrenic subjects. Total Cbl was significantly lower in older control subjects (> 60 yrs of age), primarily reflecting a >10-fold age-dependent decline in the level of MeCbl. Levels of inactive cyanocobalamin (CNCbl) were remarkably higher in fetal brain samples. In both autistic and schizophrenic subjects MeCbl and AdoCbl levels were more than 3-fold lower than age-matched controls. In autistic subjects lower MeCbl was associated with decreased MS activity and elevated levels of its substrate homocysteine (HCY). Low levels of the antioxidant glutathione (GSH) have been linked to both autism and schizophrenia, and both total Cbl and MeCbl levels were decreased in glutamate-cysteine ligase modulatory subunit knockout (GCLM-KO) mice, which exhibit low GSH levels. Thus our findings reveal a previously unrecognized decrease in brain vitamin B12 status across the lifespan that may reflect an adaptation to increasing antioxidant demand, while accelerated deficits due to GSH deficiency may contribute to neurodevelopmental and neuropsychiatric disorders.


Synergistic Killing of Polymyxin B in Combination With the Antineoplastic Drug Mitotane Against Polymyxin-Susceptible and -Resistant Acinetobacter baumannii: A Metabolomic Study.
Polymyxins are currently used as the last-resort antibiotics against multidrug-resistant Acinetobacter baumannii. As resistance to polymyxins emerges in A. baumannii with monotherapy, combination therapy is often the only remaining treatment option. A novel approach is to employ the combination of polymyxin B with non-antibiotic drugs. In the present study, we employed metabolomics to investigate the synergistic mechanism of polymyxin B in combination with the antineoplastic drug mitotane against polymyxin-susceptible and -resistant A. baumannii. The metabolomes of four A. baumannii strains were analyzed following treatment with polymyxin B, mitotane and the combination. Polymyxin B monotherapy induced significant perturbation in glycerophospholipid (GPL) metabolism and histidine degradation pathways in polymyxin-susceptible strains, and minimal perturbation in polymyxin-resistant strains. Mitotane monotherapy induced minimal perturbation in the polymyxin-susceptible strains, but caused significant perturbation in GPL metabolism, pentose phosphate pathway and histidine degradation in the LPS-deficient polymyxin-resistant strain (FADDI-AB065). The polymyxin B - mitotane combination induced significant perturbation in all strains except the lipid A modified polymyxin-resistant FADDI-AB225 strain. For the polymyxin-susceptible strains, the combination therapy significantly perturbed GPL metabolism, pentose phosphate pathway, citric acid cycle, pyrimidine ribonucleotide biogenesis, guanine ribonucleotide biogenesis, and histidine degradation. Against FADDI-AB065, the combination significantly perturbed GPL metabolism, pentose phosphate pathway, citric acid cycle, and pyrimidine ribonucleotide biogenesis. Overall, these novel findings demonstrate that the disruption of the citric acid cycle and inhibition of nucleotide biogenesis are the key metabolic features associated with synergistic bacterial killing by the combination against polymyxin-susceptible and -resistant A. baumannii.


The DEAH-box RNA helicase Dhr1 contains a remarkable carboxyl terminal domain essential for small ribosomal subunit biogenesis.
Ribosome biogenesis is an essential process in all living cells, which entails countless highly sequential and dynamic structural reorganization events. These include formation of dozens RNA helices through Watson-Crick base-pairing within ribosomal RNAs (rRNAs) and between rRNAs and small nucleolar RNAs (snoRNAs), transient association of hundreds of proteinaceous assembly factors to nascent precursor (pre-)ribosomes, and stable assembly of ribosomal proteins. Unsurprisingly, the largest group of ribosome assembly factors are energy-consuming proteins (NTPases) including 25 RNA helicases in budding yeast. Among these, the DEAH-box Dhr1 is essential to displace the box C/D snoRNA U3 from the pre-rRNAs where it is bound in order to prevent premature formation of the central pseudoknot, a dramatic irreversible long-range interaction essential to the overall folding of the small ribosomal subunit. Here, we report the crystal structure of the Dhr1 helicase module, revealing the presence of a remarkable carboxyl-terminal domain essential for Dhr1 function in ribosome biogenesis in vivo and important for its interaction with its coactivator Utp14 in vitro. Furthermore, we report the functional consequences on ribosome biogenesis of DHX37 (human Dhr1) mutations found in patients suffering from microcephaly and other neurological diseases.


Reversible effect of X-irradiation on proliferation, neurogenesis, and cell death in the dentate gyrus of adult mice.
Therapeutic cranial X-irradiation causes cognitive deficits in adult and pediatric patients, in particular, when the exposed area includes the medial temporal lobes. Effects on adult neurogenesis within the hippocampus may be related to such deficits. To investigate this relation, we irradiated the brain of young adult C57Bl/6j mice with a single dose of 4 Gy at a dose-rate of 27.5 cGy/min. We observed an approximately 80% decrease in the number of cells immunoreactive for the proliferation marker Ki67, 16 and 48 h after exposure, which was restored to control values after 1 week. The number of doublecortin- and NeuroD-immunoreactive cells of neuronal lineage was reduced by 60-70% up to 1 week after irradiation, but not after 1 month. The number of pyknotic cells increased approximately 2.5 fold after 16 h, decreased to approximately 50% of control numbers after 48 h and 1 week, and was again at normal levels after 1 month. Granule cell number did not differ between different groups and time points. There was no apparent activation of microglia or astrocytes. Our findings consist of an acute and reversible effect of X-irradiation on proliferation, neurogenesis, and cell death. Transient changes of neurogenesis may play a role in transient impairments of cognitive performance of patients exposed to X-irradiation. We present an experimental approach to temporarily alter adult hippocampal neurogenesis (AhN), allowing mechanistic investigations of AhN and its relevance to cognitive performances. The work also represents a step toward optimized radiotherapy schedules.


The Search for an Interesting Partner to Combine with PD-L1 Blockade in Mesothelioma: Focus on TIM-3 and LAG-3.
Malignant pleural mesothelioma (MPM) is an aggressive cancer that is causally associated with previous asbestos exposure in most afflicted patients. The prognosis of patients remains dismal, with a median overall survival of only 9-12 months, due to the limited effectiveness of any conventional anti-cancer treatment. New therapeutic strategies are needed to complement the limited armamentarium against MPM. We decided to focus on the combination of different immune checkpoint (IC) blocking antibodies (Abs). Programmed death-1 (PD-1), programmed death ligand-1 (PD-L1), T-cell immunoglobulin mucin-3 (TIM-3), and lymphocyte activation gene-3 (LAG-3) blocking Abs were tested as monotherapies, and as part of a combination strategy with a second IC inhibitor. We investigated their effect in vitro by examining the changes in the immune-related cytokine secretion profile of supernatant collected from treated allogeneic MPM-peripheral blood mononuclear cell (PBMC) co-cultures. Based on our in vitro results of cytokine secretion, and flow cytometry data that showed a significant upregulation of PD-L1 on PBMC after co-culture, we chose to further investigate the combinations of anti PD-L1 + anti TIM-3 versus anti PD-L1 + anti LAG-3 therapies in vivo in the AB1-HA BALB/cJ mesothelioma mouse model. PD-L1 monotherapy, as well as its combination with LAG-3 blockade, resulted in in-vivo delayed tumor growth and significant survival benefit.


High levels of transforming growth factor beta 1 correlate with disease progression in human colon cancer.
Several genes have identified that play a role in colon cancer development. However, less is known about factors that increase the rate of progression of colon cancers to metastasis. One candidate is transforming growth factor beta 1 (TGF beta 1), which can enhance the aggressiveness of human colorectal cell lines in vitro and in vivo. The amount of TGF beta 1, TGF beta 2, and TGF beta 3 protein isoforms expressed in primary site colorectal cancers were measured to determine whether any correlation existed between protein levels and disease recurrence in a series of Memorial Sloan-Kettering Hospital patients who underwent potentially curative resections. Intense staining for TGF beta 1 correlated significantly (P < 0.0013; odds ratio, 18) with disease progression to metastasis and was independent of nodal status and the degree of differentiation of the primary tumor. Therefore, in this study, patients with high TGF beta 1 protein levels in their primary site colorectal cancer were 18 times more likely to experience recurrence of their disease than were patients whose tumors exhibited low levels of TGF beta 1. In this case-control study, patients whose cancer recurred and those remaining cancer free were age and sex matched. The disease recurred at a mean of 26.8 +/- 4.3 (SE) months, whereas the mean follow-up time in patients whose disease did not recur was over twice as long, 57.3 +/- 6.6 months. Ninety-four % of the patients in each group were node positive at the time of resection, with equal mean numbers of positive nodes per patient.(ABSTRACT TRUNCATED AT 250 WORDS)


Intensive and non-intensive treatment of lexical anomia are equally efficient in post-stroke aphasia.
Although the treatment for lexical anomia in individuals with aphasia (IWA) was shown effective, little is known about the optimal treatment intensity required. The aim of this study was to verify whether intensive and non-intensive treatments led to different outcomes when parameters of intensity are rigorously controlled. Six IWA with post-stroke lexical anomia received phonological treatment at two distinct frequencies: intensive (four times a week) and non-intensive (once a week). Results showed that both treatments were equally effective. This finding is especially relevant in contexts in which speech-language therapy delivery services are limited.


Functional vascularized lung grafts for lung bioengineering.
End-stage lung disease is the third leading cause of death worldwide, accounting for 400,000 deaths per year in the United States alone. To reduce the morbidity and mortality associated with lung disease, new therapeutic strategies aimed at promoting lung repair and increasing the number of donor lungs available for transplantation are being explored. Because of the extreme complexity of this organ, previous attempts at bioengineering functional lungs from fully decellularized or synthetic scaffolds lacking functional vasculature have been largely unsuccessful. An intact vascular network is critical not only for maintaining the blood-gas barrier and allowing for proper graft function but also for supporting the regenerative cells. We therefore developed an airway-specific approach to removing the pulmonary epithelium, while maintaining the viability and function of the vascular endothelium, using a rat model. The resulting vascularized lung grafts supported the attachment and growth of human adult pulmonary cells and stem cell-derived lung-specified epithelial cells. We propose that de-epithelialization of the lung with preservation of intact vasculature could facilitate cell therapy of pulmonary epithelium and enable bioengineering of functional lungs for transplantation.


Viscous populations and their support for reciprocal cooperation.
Viscous populations (those whose members are spatially distributed and have limited mobility and locality of interaction and mating) have been proposed to support the evolution of reciprocal cooperation among self-interested individuals. Here we present a model of such a population and describe how its examination yielded the realization that different classes of viscous populations exist with differing levels of support for reciprocal cooperation. Specifically we find from our model that, in a spatially distributed population with increased viscosity, the reciprocally cooperative tit-for-tat strategy may not be globally stable due to a corresponding increase in local population density.


Catalytic mechanism of porphobilinogen synthase: the chemical step revisited by QM/MM calculations.
Porphobilinogen synthase (PBGS) catalyzes the asymmetric condensation and cyclization of two 5-aminolevulinic acid (5-ALA) substrate molecules to give porphobilinogen (PBG). The chemical step of PBGS is herein revisited using QM/MM (ONIOM) calculations. Two different protonation states and several different mechanisms are considered. Previous mechanisms based on DFT-only calculations are shown unlikely to occur. According to these new calculations, the deprotonation step rather than ring closure is rate-limiting. Both the C-C bond formation first mechanism and the C-N bond formation first mechanism are possible, depending on how the A-site ALA binds to the enzyme. We furthermore propose that future work should focus on the substrate binding step rather than the enzymatic mechanism.


Tongue biofilm areal density and tongue coating index.
Two methods have been employed to measure the amount of tongue biofilm in humans: (1) areal density of microbes (cfu cm(-2)) (tongue scrape from a known/measured area), or (2) a tongue-coating index, where a human judge assesses the amount of 'coating' by visual inspection. The two methods were compared, both on human volunteers and on artificial biofilms. Fusobacterium nucleatum (Fn), Prevotella intermedia (Pi), Streptococcus mutans (Sm) and Peptostreptococcus micros (Pm) were obtained from stock cultures. Tongue scrape samples were obtained from four human volunteers in order to inoculate the in vitro matrices. A further 16 individuals (total n = 20) were co-measured for both visible tongue coating and quantitative biofilm (cfu cm(-2)). Tongue scrape suspensions were used to inoculate a cellulose matrix resting on a BHI-blood agar plate, periodically hydrated with sterile BHI and incubated for 10 days. Visual coating density images were recorded by camera. The relative light absorbance of standard suspensions of cells (pure species) was obtained by spectrophotometry across the visible spectrum. Biomatrix yields (log cfu cm(-2) ± SD) for Fn, Pi, Sm and Pm were 7.07 (±0.04), 7.38 (±0.07), 7.25 (±0.10) and 7.41 (±0.011). Higher yields were obtained for tongue-scrape mixed cultures, ranging from 7.47 to 7.76. Biofilms (in vitro and in vivo) had different appearances when measured by human judges in terms of colour, pigmentation and opacity. There was no correlation (R(2) < 0.02) between the two methods. Biofilm matrices of similar population densities (ca ∼ log 7.5) had widely different opacities depending on the species present.


Investigation of the pathophysiological mechanisms of migraine attacks induced by pituitary adenylate cyclase-activating polypeptide-38.
Pituitary adenylate cyclase-activating polypeptide-38 (PACAP38) and vasoactive intestinal polypeptide are structurally and functionally closely related but show differences in migraine-inducing properties. Mechanisms responsible for the difference in migraine induction are unknown. Here, for the first time, we present a head-to-head comparison study of the immediate and long-lasting observations of the migraine-inducing, arterial, physiological and biochemical responses comparing PACAP38 and vasoactive intestinal polypeptide. In a double-blind crossover study 24 female migraine patients without aura were randomly allocated to intravenous infusion of PACAP38 (10 pmol/kg/min) or vasoactive intestinal polypeptide (8 pmol/kg/min) over 20 min. We recorded incidence of migraine during and after infusion (0-24 h). Magnetic resonance angiography of selected extra- and intracranial arteries, blood samples (plasma PACAP38 and vasoactive intestinal polypeptide and serum tryptase), and vital signs (blood pressure, heart rate, respiratory frequency, and end-tidal pressure of CO2) was recorded before and up to 5 h after infusion. Twenty-two patients [mean age 24 years (range 19-36)] completed the study on both days. Sixteen patients (73%) reported migraine-like attacks after PACAP38 and four after vasoactive intestinal polypeptide (18%) infusion (P = 0.002). Three of four patients, who reported migraine-like attacks after vasoactive intestinal polypeptide, also reported attacks after PACAP38. Both peptides induced marked dilatation of the extracranial (P < 0.05), but not intracranial arteries (P > 0.05). PACAP38-induced vasodilatation was longer lasting (>2 h), whereas vasoactive intestinal polypeptide-induced dilatation was normalized after 2 h. We recorded elevated plasma PACAP38 at 1 h after the start of PACAP38 infusion only in those patients who later reported migraine attacks. Blood levels of vasoactive intestinal polypeptide and tryptase were unchanged after PACAP38 infusion. In conclusion, PACAP38-induced migraine was associated with sustained dilatation of extracranial arteries and elevated plasma PACAP38 before onset of migraine-like attacks. PACAP38 has a much higher affinity for the PAC1 receptor and we therefore suggest that migraine induction by PACAP38 may be because of activation of the PAC1 receptor, which may be a future anti-migraine drug target.


The integrated care of asthma in Switzerland (INCAS)-study: Patients' perspective of received asthma care and their interest in asthma education.
For successful long-term asthma care, self-management education is a cornerstone. Little is known about associations between patients' interest in education, asthma control and care delivery. We compared patients' characteristics, asthma control and patients' perspective about asthma care in subjects with and without interest in asthma education. Moreover, we assessed reasons, why patients denied participating in asthma education. Baseline data of 223 patients with asthma (age 43 ± 12 years, 38% male, 58% non-smokers, 13% current smokers), who participated in a multicentre longitudinal controlled study, are reported. At baseline, patients completed the Asthma Control Test (ACT), the Patient Assessment Chronic Illness Care questionnaire (PACIC 5A) and stated their interest in an asthma education programme. Overall, 34% of all participants showed uncontrolled asthma. One hundred and twenty-five (56%) patients were interested in education. Compared to patients without interest, they were characterised by male gender (p = 0.013), worse asthma control (p < 0.001), and perception of lower quality of chronic asthma care delivery, in particular lower self-management support (p < 0.001). Main reasons for rejecting asthma education were having sufficient asthma knowledge, having only mild asthma, receiving adequate medical support and lack of time. More than half of the patients were interested in asthma education. Interest was associated with worse asthma control and lower receipt of care according to the Chronic Care Model. Considering these aspects, this approach may help to improve care quality and allow targeting interventions to those patients who are interested in becoming active participants in their care and who might benefit most.


The role of veterans affairs in emergency management: a systematic literature review.
The Department of Veterans Affairs (VA) is a key player in emergency management for both veterans and civilians. Unfortunately, limited evidence-based research findings exist regarding the role and experience of VA during disasters. The present study is a systematic literature review of 41 published, peer-reviewed articles regarding VA and emergency management. Trained researchers utilized a data abstraction tool and conducted a qualitative content analysis. A description of article characteristics include methodology, phase of emergency management addressed in the research, and study design. Five topic categories emerged from the review including effects of disaster on mental health status and services use, effects of disaster on general health services use, patient tracking, evacuation, and disaster planning/preparation. Findings were used to generate suggestions for future research. Veterans Affairs, veterans, disaster, emergency.


Periocular corticosteroid injections in uveitis: effects and complications.
To evaluate the benefits and complications of periocular depot corticosteroid injections in patients with ocular inflammatory disorders. Multicenter, retrospective cohort study. A total of 914 patients (1192 eyes) who had received ≥ 1 periocular corticosteroid injection at 5 tertiary uveitis clinics in the United States. Patients were identified from the Systemic Immunosuppressive Therapy for Eye Diseases Cohort Study. Demographic and clinical characteristics were obtained at every visit via medical record review by trained reviewers. Control of inflammation, improvement of visual acuity (VA) to ≥ 20/40, improvement of VA loss attributed to macular edema (ME), incident cataract affecting VA, cataract surgery, ocular hypertension, and glaucoma surgery. Among 914 patients (1192 eyes) who received ≥ 1 periocular injection during follow-up, 286 (31.3%) were classified as having anterior uveitis, 303 (33.3%) as intermediate uveitis, and 324 (35.4%) as posterior or panuveitis. Cumulatively by ≤ 6 months, 72.7% (95% CI, 69.1-76.3) of the eyes achieved complete control of inflammation and 49.7% (95% CI, 45.5-54.1) showed an improvement in VA from <20/40 to ≥ 20/40. Among the subset with VA <20/40 attributed to ME, 33.1% (95% CI, 25.2-42.7) improved to ≥ 20/40. By 12 months, the cumulative incidence of ≥ 1 visits with an intraocular pressure of ≥ 24 mmHg and ≥ 30 mmHg was 34.0% (95% CI, 24.8-45.4) and 15.0% (95% CI, 11.8-19.1) respectively; glaucoma surgery was performed in 2.4% of eyes (95% CI, 1.4-3.9). Within 12 months, among phakic eyes initially ≥ 20/40, the incidence of a reduction in VA to <20/40 attributed to cataract was 20.2% (95% CI, 15.9-25.6); cataract surgery was performed within 12 months in 13.8% of the initially phakic eyes (95% CI, 11.1-17.2). Periocular injections were effective in treating active intraocular inflammation and in improving reduced VA attributed to ME in a majority of patients. The response pattern was similar across anatomic locations of uveitis. Overall, VA improved in one half of the patients at some point within 6 months. However, cataract and ocular hypertension occurred in a substantial minority.


Influence of socio-economic status on habitual physical activity and sedentary behavior in 8- to 11-year old children.
While socio-economic status has been shown to be an important determinant of health and physical activity in adults, results for children and adolescents are less consistent. The purpose of this study, therefore, is to examine whether physical activity and sedentary behavior differs in children by socio-economic status (SES) independent of body mass index. Data were from two cohorts including 271 children (117 males; 154 females) in study 1 and 131 children in study 2 (63 males; 68 females). The average age was 9.6 and 8.8 years respectively. Height and body mass were assessed according to standard procedures and body mass index (BMI, kg/m2) was calculated. Parent-reported household income was used to determine SES. Habitual, free-living physical activity (PA) was assessed by a pedometer (steps/day) in study 1 and accelerometer (time spent in moderate-to-vigorous PA) in study 2. Self-reported time spent watching TV and on the computer was used as measure of sedentary behavior. Differences in PA and sedentary behavior by SES were initially tested using ANOVA. Further analyses used ANCOVA controlling for BMI, as well as leg length in the pedometer cohort. In study 1, mean daily steps differed significantly among SES groups with lower SES groups approximating 10,500 steps/day compared to about 12,000 steps/day in the higher SES groups. These differences remained significant (p < 0.05) when controlling for leg length. Lower SES children, however, had higher body mass and BMI compared to higher SES groups (p < 0.05) and PA no longer remained significant when further controlling for BMI. In study 2 results depended on the methodology used to determine time spent in moderate-to-vigorous physical activity (MVPA). Only one equation resulted in significant group differences (p = 0.015), and these differences remained after controlling for BMI. Significant differences between SES groups were shown for sedentary behavior in both cohorts (P < 0.05) with higher SES groups spending less time watching TV than low SES groups. Children from a low SES show a trend of lower PA levels and spend more time in sedentary behavior than high SES children; however, differences in PA were influenced by BMI. The higher BMI in these children might be another factor contributing to increased health risks among low SES children compared to children from with a higher SES.


Flavodiiron-Mediated O2 Photoreduction Links H2 Production with CO2 Fixation during the Anaerobic Induction of Photosynthesis.
Some microalgae, such as Chlamydomonas reinhardtii, harbor a highly flexible photosynthetic apparatus capable of using different electron acceptors, including carbon dioxide (CO2), protons, or oxygen (O2), allowing survival in diverse habitats. During anaerobic induction of photosynthesis, molecular O2 is produced at photosystem II, while at the photosystem I acceptor side, the reduction of protons into hydrogen (H2) by the plastidial [FeFe]-hydrogenases primes CO2 fixation. Although the interaction between H2 production and CO2 fixation has been studied extensively, their interplay with O2 produced by photosynthesis has not been considered. By simultaneously measuring gas exchange and chlorophyll fluorescence, we identified an O2 photoreduction mechanism that functions during anaerobic dark-to-light transitions and demonstrate that flavodiiron proteins (Flvs) are the major players involved in light-dependent O2 uptake. We further show that Flv-mediated O2 uptake is critical for the rapid induction of CO2 fixation but is not involved in the creation of the micro-oxic niches proposed previously to protect the [FeFe]-hydrogenase from O2 By studying a mutant lacking both hydrogenases (HYDA1 and HYDA2) and both Flvs (FLVA and FLVB), we show that the induction of photosynthesis is strongly delayed in the absence of both sets of proteins. Based on these data, we propose that Flvs are involved in an important intracellular O2 recycling process, which acts as a relay between H2 production and CO2 fixation.


[Villitis of unknown etiology in placentas of pregnancies with hypertensive disorders and of small-for-gestational-age infants].
The objectives of this study are to analyze the frequency and the histopathological features of the villitis of unknown etiology (VUE) in placentas of pregnancies with hypertensive disorders and of small-for-gestational-age infants (SGA). Two hundred and thirteen placentas from pregnancies without clinical or laboratorial evidence of infection were studied. These cases were subdivided according to: a) maternal condition in: non-complicated pregnancy (NCP)- 151 cases, pregnancy-induced hypertension (PIH)- 37 and sustained chronic hypertension (SCH)- 25 and b) newborn weight in: small for gestational age (SGA)- 38 cases and adequate for gestational age (AGA)- 175. Of these placentas, 81 belong to the random sample, which was used to determine the frequency of VUE in the population studied. Eight blocks were taken from placental parenchyma and the histological sections were stained with hematoxylin and eosin. Frequency tables of categorical data were analyzed using the chi- square test and Fisher test; statistical significance was considered for p< 0.05. The frequency of VUE in the placentas was 30.8% in the random sample, 39% in NCP, 29.7% in PIH, 32% in SCH, 34.2% in SGA infants and 37.1% in AGA infants. Placentas of pregnancies with hypertensive disorders were more affected by villitis with parenchymatous component (PIH - 27.0%, SCH - 28.0%). This lesion was also the predominant villitis in the placentas of the SGA infants (31.5%). In contrast, in placentas of NCP and AGA infants, the principal kind of villitis was basal, not associated to a parenchymatous component (16.5% and 14.8% respectively). In the majority of the cases the villitis was mild. In the population studied, the frequency of VUE is high (around 30%). The lesion occurs in a similar frequency in placentas from NCP, PIH, SCH, SGA and AGA infants, but basal villitis not associated to a parenchymatous component affects mainly the placentas of NCP and AGA infants. It is possible that this kind of villitis could be an expression of an antigenic stimulation diverse from villitis with a parenchymatous component.


Long-term Satellite NDVI Data Sets: Evaluating Their Ability to Detect Ecosystem Functional Changes in South America.
In the last decades, South American ecosystems underwent important functional modifications due to climate alterations and direct human intervention on land use and land cover. Among remotely sensed data sets, NOAA-AVHRR "Normalized Difference Vegetation Index" (NDVI) represents one of the most powerful tools to evaluate these changes thanks to their extended temporal coverage. In this paper we explored the possibilities and limitations of three commonly used NOAA-AVHRR NDVI series (PAL, GIMMS and FASIR) to detect ecosystem functional changes in the South American continent. We performed pixel-based linear regressions for four NDVI variables (average annual, maximum annual, minimum annual and intra-annual coefficient of variation) for the 1982-1999 period and (1) analyzed the convergences and divergences of significant multi-annual trends identified across all series, (2) explored the degree of aggregation of the trends using the O-ring statistic, and (3) evaluated observed trends using independent information on ecosystem functional changes in five focal regions. Several differences arose in terms of the patterns of change (the sign, localization and total number of pixels with changes). FASIR presented the highest proportion of changing pixels (32.7%) and GIMMS the lowest (16.2%). PAL and FASIR data sets showed the highest agreement, with a convergence of detected trends on 71.2% of the pixels. Even though positive and negative changes showed substantial spatial aggregation, important differences in the scale of aggregation emerged among the series, with GIMMS showing the smaller scale (≤11 pixels). The independent evaluations suggest higher accuracy in the detection of ecosystem changes among PAL and FASIR series than with GIMMS, as they detected trends that match expected shifts. In fact, this last series eliminated most of the long term patterns over the continent. For example, in the "Eastern Paraguay" and "Uruguay River margins" focal regions, the extensive changes due to land use and land cover change expansion were detected by PAL and FASIR, but completely ignored by GIMMS. Although the technical explanation of the differences remains unclear and needs further exploration, we found that the evaluation of this type of remote sensing tools should not only be focused at the level of assumptions (i.e. physical or mathematical aspects of image processing), but also at the level of results (i.e. contrasting observed patterns with independent proofs of change). We finally present the online collaborative initiative "Land ecosystem change utility for South America", which facilitates this type of evaluations and helps to identify the most important functional changes of the continent.


Translocation of silica nanospheres through giant unilamellar vesicles (GUVs) induced by a high frequency electromagnetic field.
Membrane model systems capable of mimicking live cell membranes were used for the first time in studying the effects arising from electromagnetic fields (EMFs) of 18 GHz where membrane permeability was observed following exposure. A present lack of understanding of the mechanisms that drive such a rapid change in membrane permeabilization as well as any structural or dynamic changes imparted on biomolecules affected by high-frequency electromagnetic irradiation limits the use of 18 GHz EMFs in biomedical applications. A phospholipid, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) labelled with a fluorescent marker 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (rhodamine-DOPE) was used in constructing the giant unilamellar vesicles (GUVs). After three cycles of exposure, enhanced membrane permeability was observed by the internalisation of hydrophilic silica nanospheres of 23.5 nm and their clusters. All-atom molecular dynamics simulations of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes exposed to high frequency electric fields of different field strengths showed that within the simulation timeframe only extremely high strength fields were able to cause an increase in the interfacial water dynamics characterized by water dipole realignments. However, a lower strength, high frequency EMF induced changes of the water hydrogen bond network, which may contribute to the mechanisms that facilitate membrane permeabilization in a longer timeframe.


Experiences of working with the tobacco issue in the context of health promoting hospitals and health services: a qualitative study.
The worldwide Health Promoting Hospital and Health Services (HPH) network was initiated by the World Health Organizations in the late 1980s. The goal of the network is to change the focus of health services from curing patients to also embrace disease prevention and health promotion. In Sweden the network started in 1996, and involves mainly hospitals and primary care. The network members collaborate in task forces, one of which is working on the tobacco issue. There is limited evidence on the value of working within an HPH organization. The aim of this study was to investigate the experiences of members of the Swedish HPH network tobacco task force. Focus group interviews with task force members were analyzed using implementation theory. Three themes, overall experiences of working with tobacco issues, experiences of working with "free from tobacco in connection with surgery", and experiences of work in the HPH tobacco task force, emerged from the interviews. The results show that working with the tobacco issue in the context of health-promoting hospitals and health services met with difficulties involving the following important factors: evidence, context, facilitation and adopter characteristics. Leadership, one contextual factor, at national and local level, seems to be crucial if the work is going to succeed. The tobacco task force of the HPH network is an important facilitator supporting the task.