Identification of new developmentally regulated genes involved in Streptomyces coelicolor sporulation.
The sporulation of aerial hyphae of Streptomyces coelicolor is a complex developmental process. Only a limited number of the genes involved in this intriguing morphological differentiation programme are known, including some key regulatory genes. The aim of this study was to expand our knowledge of the gene repertoire involved in S. coelicolor sporulation. We report a DNA microarray-based investigation of developmentally controlled gene expression in S. coelicolor. By comparing global transcription patterns of the wild-type parent and two mutants lacking key regulators of aerial hyphal sporulation, we found a total of 114 genes that had significantly different expression in at least one of the two mutants compared to the wild-type during sporulation. A whiA mutant showed the largest effects on gene expression, while only a few genes were specifically affected by whiH mutation. Seven new sporulation loci were investigated in more detail with respect to expression patterns and mutant phenotypes. These included SCO7449-7451 that affect spore pigment biogenesis; SCO1773-1774 that encode an L-alanine dehydrogenase and a regulator-like protein and are required for maturation of spores; SCO3857 that encodes a protein highly similar to a nosiheptide resistance regulator and affects spore maturation; and four additional loci (SCO4421, SCO4157, SCO0934, SCO1195) that show developmental regulation but no overt mutant phenotype. Furthermore, we describe a new promoter-probe vector that takes advantage of the red fluorescent protein mCherry as a reporter of cell type-specific promoter activity. Aerial hyphal sporulation in S. coelicolor is a technically challenging process for global transcriptomic investigations since it occurs only as a small fraction of the colony biomass and is not highly synchronized. Here we show that by comparing a wild-type to mutants lacking regulators that are specifically affecting processes in aerial hypha, it is possible to identify previously unknown genes with important roles in sporulation. The transcriptomic data reported here should also serve as a basis for identification of further developmentally important genes in future functional studies.


Risk Stratification of Pleural Fluid Cytology Based on the International System for Reporting Serous Fluid Cytology in a Tertiary Care Centre.
The International Academy of Cytology and the American Society of Cytopathology proposed the international system for reporting serous fluid cytology (TISRSFC) in 2019 to form uniform reporting terminologies for fluid cytology. This system defines a spectrum of diagnostic categories to be used in daily clinical practice. Fluid cytology is a widely accepted, cost-effective, minimally invasive earliest diagnostic method for the investigation that aids management decisions. The present study aims to reclassify pleural fluid cytology and calculate the risk of malignancy (ROM) for each diagnostic category. It is a retrospective 2 years observational study comprising 690 pleural fluid specimens. Retrospective reviews were performed, and cases were reclassified into five categories as per TISRSFC. Immunohistochemistry was applied whenever needed. Cytological diagnosis was correlated with respective histopathology and/or clinical and/or radiological diagnosis. ROM was calculated for all diagnostic categories. In the present study of 690 cases, 7.97% were non-diagnostic (ND), 84.1% cases were negative for malignancy (NFM), 0.87% were atypia of undetermined significance (AUS), 1.16% were suspicious for malignancy (SFM), and 5.94% were malignant (MAL). Cell blocks were prepared in 33 (4.8%) cases and immunohistochemistry was applied in 7 cases. The commonest site for pleural fluid metastasis was the lungs, accounting for 4.64% (32/690) cases. Further, ROM was calculated for all diagnostic categories as follows: (1) ND: 30.9%, (2) NFM: 12.9%, (3) AUS: 100%, (4) SFM: 100%, and (5) MAL: 90.2%. Cytological examination of pleural fluids is an accurate, prompt, and affordable technique. This standardized ISRSFC reporting system will maintain uniformity and reproducibility in reporting, leading to improved clinical decision-making of pleural fluids.


Incorporation of Lippia citriodora Microwave Extract into Total-Green Biogelatin-Phospholipid Vesicles to Improve Its Antioxidant Activity.
Phytochemicals from Lippia citriodora leaves were extracted by applying an innovative technology based on the use of microwaves, which represents an alternative method to extract bioactive substances. The obtained extract was incorporated into phospholipid vesicles in order to promote the antioxidant effect of the bioactive molecules present in L. citriodora extract. The extract was analyzed by High Performance Liquid Chromatography coupled to Time-Of-Flight mass spectrometer by electrospray (HPLC-ESI-TOF-MS) and different phytochemicals were detected and quantified. The whole extract was incorporated in liposomes, glycerosomes (liposomes modified with glycerol) and propylene glycol-containing vesicles (PG-PEVs). Moreover, a biopolymer obtained from fish by-product, that is Thunnus albacares skin, was added to improve the bioactivity of the formulations. The in vitro biocompatibility and the antioxidant efficacy of the extract in solution or loaded in the vesicles were tested in primary mouse embryonic fibroblasts (3T3). The results showed the superior bioactivity of the vesicle formulations over the aqueous solution of the extract, which points to an interesting strategy for the treatment of skin disorders.


Impact of public health emergencies on modern disaster taxonomy, planning, and response.
Current disaster taxonomy describes diversity, distinguishing characteristics, and common relations in disaster event classifications. The impact of compromised public health infrastructure and systems on health consequences defines and greatly influences the manner in which disasters are observed, planned for, and managed, especially those that are geographically widespread, population dense, and prolonged. What may first result in direct injuries and death may rapidly change to excess indirect illness and subsequent death as essential public health resources are destroyed, deteriorate, or are systematically denied to vulnerable populations. Public health and public health infrastructure and systems in developed and developing countries must be seen as strategic and security issues that deserve international public health resource monitoring attention from disaster managers, urban planners, the global humanitarian community, World Health Organization authorities, and participating parties to war and conflict. We posit here that disaster frameworks be reformed to emphasize and clarify the relation of public health emergencies and modern disasters.


Learning to Use an Alphabetic Writing System.
Gaining facility with spelling is an important part of becoming a good writer. Here we review recent work on how children learn to spell in alphabetic writing systems. Statistical learning plays an important role in this process. Thus, young children learn about some of the salient graphic characteristics of written texts and attempt to reproduce these characteristics in their own productions even before they use letters to represent phonemes. Later, children apply their statistical learning skills to links between phonemes and spellings, including those that are conditioned by context and morphology. Children use what they know about language and about letter names when learning about spelling, and learning to spell in turn influences their ideas about language. Although children learn about some aspects of spelling implicitly, explicit instruction has an important role to play. We discuss some implications of the research for the design of that instruction.


R-Ras subfamily proteins elicit distinct physiologic effects and phosphoproteome alterations in neurofibromin-null MPNST cells.
Loss of the Ras GTPase-activating protein neurofibromin promotes nervous system tumor pathogenesis in patients with neurofibromatosis type 1 (NF1). Neurofibromin loss potentially hyperactivates classic Ras (H-Ras, N-Ras, K-Ras), M-Ras, and R-Ras (R-Ras, R-Ras2/TC21) subfamily proteins. We have shown that classic Ras proteins promote proliferation and survival, but not migration, in malignant peripheral nerve sheath tumor (MPNST) cells. However, it is unclear whether R-Ras, R-Ras2 and M-Ras are expressed and hyperactivated in MPNSTs and, if so, whether they contribute to MPNST pathogenesis. We assessed the expression and activation of these proteins in MPNST cells and inhibited them to determine the effect this had on proliferation, migration, invasion, survival and the phosphoproteome. NF1-associated (ST88-14, 90-8, NMS2, NMS-PC, S462, T265-2c) and sporadic (STS-26T, YST-1) MPNST lines were used. Cells were transfected with doxycycline-inducible vectors expressing either a pan-inhibitor of the R-Ras subfamily [dominant negative (DN) R-Ras] or enhanced green fluorescent protein (eGFP). Methodologies used included immunoblotting, immunocytochemistry, PCR, Transwell migration, 3H-thymidine incorporation, calcein cleavage assays and shRNA knockdowns. Proteins in cells with or without DN R-Ras expression were differentially labeled with SILAC and mass spectrometry was used to identify phosphoproteins and determine their relative quantities in the presence and absence of DN R-Ras. Validation of R-Ras and R-Ras2 action and R-Ras regulated networks was performed using genetic and/or pharmacologic approaches. R-Ras2 was uniformly expressed in MPNST cells, with R-Ras present in a major subset. Both proteins were activated in neurofibromin-null MPNST cells. Consistent with classical Ras inhibition, DN R-Ras and R-Ras2 knockdown inhibited proliferation. However, DN R-Ras inhibition impaired migration and invasion but not survival. Mass spectrometry-based phosphoproteomics identified thirteen protein networks distinctly regulated by DN R-Ras, including multiple networks regulating cellular movement and morphology. ROCK1 was a prominent mediator in these networks. DN R-Ras expression and RRAS and RRAS2 knockdown inhibited migration and ROCK1 phosphorylation; ROCK1 inhibition similarly impaired migration and invasion, altered cellular morphology and triggered the accumulation of large intracellular vesicles. R-Ras proteins function distinctly from classic Ras proteins by regulating distinct signaling pathways that promote MPNST tumorigenesis by mediating migration and invasion. Mutations of the NF1 gene potentially results in the activation of multiple Ras proteins, which are key regulators of many biologic effects. The protein encoded by the NF1 gene, neurofibromin, acts as an inhibitor of both classic Ras and R-Ras proteins; loss of neurofibromin could cause these Ras proteins to become persistently active, leading to the development of cancer. We have previously shown that three related Ras proteins (the classic Ras proteins) are highly activated in malignant peripheral nerve sheath tumor (MPNST) cells with neurofibromin loss and that they drive cancer cell proliferation and survival by activating multiple cellular signaling pathways. Here, we examined the expression, activation and action of R-Ras proteins in MPNST cells that have lost neurofibromin. Both R-Ras and R-Ras2 are expressed in MPNST cells and activated. Inhibition of R-Ras action inhibited proliferation, migration and invasion but not survival. We examined the activation of cytoplasmic signaling pathways in the presence and absence of R-Ras signaling and found that R-Ras proteins regulated 13 signaling pathways distinct from those regulated by classic Ras proteins. Closer study of an R-Ras regulated pathway containing the signaling protein ROCK1 showed that inhibition of either R-Ras, R-Ras2 or ROCK1 similarly impaired cellular migration and invasion and altered cellular morphology. Inhibition of R-Ras/R-Ras2 and ROCK1 signaling also triggered the accumulation of abnormal intracellular vesicles, indicating that these signaling molecules regulate the movement of proteins and other molecules in the cellular interior. Video Abstract.


Strain-encoded CMR for the detection of inducible ischemia during intermediate stress.
This study sought to evaluate the diagnostic accuracy of strain-encoded cardiac magnetic resonance (SENC) for the detection of inducible ischemia during intermediate stress. High-dose dobutamine stress cardiac magnetic resonance (DS-CMR) is a well-established modality for the noninvasive detection of coronary artery disease (CAD). However, the assessment of cine scans relies on the visual interpretation of wall motion, which is subjective, and modalities that can objectively and quantitatively assess the time course of myocardial strain response during stress are lacking. Stress-induced ischemia was assessed by wall motion analysis and by SENC in 80 patients with suspected or known CAD and in 18 healthy volunteers who underwent DS-CMR in a clinical 1.5-T scanner. Quantitative coronary angiography was used as the standard reference for the presence of CAD (> or =50% diameter stenosis). On a patient level, 46 of 80 patients (58%) had CAD, including 20 with single-vessel, 18 with 2-vessel, and 8 with 3-vessel disease. During peak stress, SENC correctly detected ischemia in 45 versus 38 of 46 patients with CAD (7 additional correct findings for SENC), yielding significantly higher sensitivity than cine (98% vs. 83%, p < 0.05). No patients were correctly diagnosed by cine and missed by SENC. During intermediate stress, SENC showed diagnostic value similar to that provided by cine imaging only during peak dobutamine stress (sensitivity of 76% vs. 83%, specificity of 88% vs. 91%, and accuracy of 81% vs. 86%; p = NS for all). Quantification analysis demonstrated that strain rate response is a highly sensitive marker for the detection of inducible ischemia (area under the curve = 0.96; SE = 0.01; 95% confidence interval: 0.93 to 0.99) that precedes the development of inducible wall motion abnormalities and already significantly decreases with moderate 40% to 60% coronary lesions. Using SENC, CAD can be detected during intermediate stress with similar accuracy to that provided by cine only during peak stress. By this approach, patient safety may be improved during diagnostic procedures within lower time spent (Strain-Encoded Cardiac Magnetic Resonance Imaging for Dobutamine Stress Testing; NCT00758654).


Spirometry for the diagnosis of airway obstruction in patients with risk factors for COPD: the GOLD and lower limit of normal criteria.
The identification of persistent airway obstruction is key to making a diagnosis of COPD. The GOLD guidelines suggest a fixed criterion-a post-bronchodilator FEV1/FVC ratio < 70%-to define obstruction, although other guidelines suggest that a post-bronchodilator FEV1/FVC ratio < the lower limit of normal (LLN) is the most accurate criterion. This was an observational study of individuals ≥ 40 years of age with risk factors for COPD who were referred to our pulmonary function laboratory for spirometry. Respiratory symptoms were also recorded. We calculated the prevalence of airway obstruction and of no airway obstruction, according to the GOLD criterion (GOLD+ and GOLD-, respectively) and according to the LLN criterion (LLN+ and LLN-, respectively). We also evaluated the level of agreement between the two criteria. A total of 241 individuals were included. Airway obstruction was identified according to the GOLD criterion in 42 individuals (17.4%) and according to the LLN criterion in 23 (9.5%). The overall level of agreement between the two criteria was good (k = 0.67; 95% CI: 0.52-0.81), although it was lower among the individuals ≥ 70 years of age (k = 0.42; 95% CI: 0.12-0.72). The proportion of obese individuals was lower in the GOLD+/LLN+ category than in the GOLD+/LLN- category (p = 0.03), as was the median DLCO (p = 0.04). The use of the GOLD criterion appears to be associated with a higher prevalence of COPD. The agreement between the GOLD and LLN criteria also appears to be good, albeit weaker in older individuals. The use of different criteria to define airway obstruction seems to identify individuals with different characteristics. It is essential to understand the clinical meaning of discordance between such criteria. Until more data are available, we recommend a holistic, individualized approach to, as well as close follow-up of, patients with discordant results for airway obstruction.


Total Burden of Cerebral Small Vessel Disease in Recurrent ICH versus First-ever ICH.
The relationship between recurrent intracerebral hemorrhage (ICH) and total burden of cerebral small vessel disease (CSVD) is not completely investigated. We aimed to study whether recurrent intracerebral hemorrhage (ICH) had higher CSVD score than first-ever ICH. Lacunes, white matter hyperintensities (WMH), cerebral microbleeds (CMBs), enlarged perivascular spaces (EPVS), cortical superficial siderosis (cSS) and CSVD score were rated on brain magnetic resonance imaging (MRI) in primary ICH patients. Recurrent ICHs were confirmed by reviewing the medical records and MRI scans. Mixed hematomas were defined as follows: deep + lobar, deep + cerebellar, or deep + lobar + cerebellar. Of the 184 patients with primary ICH enrolled (mean age, 61.0 years; 75.5% men), recurrent ICH was present in 45 (24.5%) patients; 26.1% (48/184) had ≥2 hematomas, 93.8% (45/48) of which exhibited recurrent ICH. Mixed hematomas were identified in 8.7% (16/184) of patients and bilateral hematomas in 17.9% (33/184). All mixed hematomas and bilateral hematomas were from cases of recurrent ICH. Patients with mixed etiology-ICH were more likely to have recurrent ICH than patients with cerebral amyloid angiopathy (CAA) or hypertensive angiopathy (HA)-related ICH (36.8% vs17.8%, p=0.008). Multivariate ordinal regression analysis showed that the presence of recurrent ICH (p=0.001), ≥2 hematomas (p=0.002), mixed hematomas (p<0.00001), and bilateral hematomas (p=0.002) were separately significantly associated with a high CSVD score. Recurrent ICH occurs mostly among patients with mixed etiology-ICH and is associated with a higher CSVD burden than first-ever ICH, which needs to be verified by future larger studies.


Thoracic aortic involvement in cystic echinococcosis: A case report.
Cystic echinococcosis is a common parasitic disease in some areas and involves different organs with different presentations. Rural areas in our country are endemic with this disease. We present a case of echinococcal aortic involvement that was diagnosed incidentally by a routine chest radiogram as a mediastinal mass. Cystic echinococcosis was established as final diagnosis after magnetic resonance imaging (MRI) and ELISA studies. The patient was treated surgically through a thoracotomy and her aortic defect was repaired during the operation. She has been asymptomatic and complication free in physical examination and imaging study in follow up periods after two years. Awareness of various presentations of the disease is necessary for physicians who are occupied in endemic areas. It is rational to treat echinococcal cysts adjacent to the great vessels with the cooperation of a vascular surgeon.


Understanding Care Navigation by Older Adults With Multimorbidity: Mixed-Methods Study Using Social Network and Framework Analyses.
Health and social care systems were designed to be used primarily by people with single and acute diseases. However, a growing number of older adults are diagnosed with multiple long-term conditions (LTCs). The process of navigating the intricacies of health and social care systems to receive appropriate care presents significant challenges for older people living with multiple LTCs, which in turn can negatively influence their well-being and quality of life. The long-term goal of this work is to design technology to assist people with LTCs in navigating health and social care systems. To do so, we must first understand how older people living with LTCs currently engage with and navigate their care networks. No published research describes and analyses the structure of formal and informal care networks of older adults with multiple LTCs, the frequency of interactions with each type of care service, and the problems that typically arise in these interactions. We conducted a mixed-methods study and recruited 62 participants aged ≥55 years who were living in England, had ≥2 LTCs, and had completed a social network analysis questionnaire. Semistructured interviews were conducted with roughly a 10% subsample of the questionnaire sample: 4 women and 3 men. On average, interviewees aged 70 years and had 4 LTCs. Personal care networks were complex and adapted to each individual. The task of building and subsequently navigating one's personal care network rested mainly on patients' shoulders. It was frequently the patients' task to bridge and connect the different parts of the system. The major factor leading to a satisfying navigation experience was found to be patients' assertive, determined, and proactive approaches. Furthermore, smooth communication and interaction between different parts of the care system led to more satisfying navigation experiences. Technology to support care navigation for older adults with multiple LTCs needs to support patients in managing complex health and social care systems by effectively integrating the management of multiple conditions and facilitating communication among multiple stakeholders, while also offering flexibility to adapt to individual situations. Quality of care seems to be dependent on the determination and ability of patients. Those with less determination and fewer organization skills experience worse care. Thus, technology must aim to fulfill these coordination functions to ensure care is equitable across those who need it.


Exploratory laparotomy in the detection of abdominal metastases of primary lung cancer.
From January 1974 to December 1976 122 exploratory laparotomies were performed in patients with primary lung cancer. With this method detection of asymptomatic abdominal metastases was achieved in 23.8% of the cases. The more frequent sites of metastases were the abdominal lymph nodes and the liver. The most common histological findings were oat-cell carcinoma (50%) and adenocarcinoma (35.7%). The detection of asymptomatic abdominal metastases using this procedure permitted the modification of the clinical staging (adopted by the UICC) in 9.8% of the cases, with emphasis on clinical stage I and II. This diagnostic procedure is harmless: no patient died during the period of the study. The authors recommended inclusion of exploratory laparotomy as an adequate routine means both for the detection of abdominal metastases and for the surgical and clinical staging of primary lung cancer, with emphasis on female patients and clinical stages I and II.


Interleukin-10 may participate in regulating trophoblast invasion in human placentae throughout gestation.
A successful human pregnancy requires cytotrophoblasts from the fetal portion of the placenta to adopt tumor-like properties. But unlike tumor metastasis, cytotrophoblast invasion is highly regulated both spatially and temporally. The mechanisms that regulate human trophoblast invasion are understood poorly. With a view to obtain some findings on the mechanisms that regulate human trophoblast invasion, we applied the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) method to compare the expression of invasion-associated genes in cytotrophoblasts isolated from first- and third-trimester placental tissues. In trophoblast cells of first-trimester pregnancy, the mRNA contents of matrix metalloproteinase (MMP)-9 and urokinase-type plasminogen activator (uPA) were higher than that in the third-trimester cytotrophoblasts, while no difference of MMP-2 mRNA expression level was found between trophoblastic cells of different gestational ages. The expression level of plasminogen activator inhibitors-1 mRNA in first-trimester cytotrophoblasts was shown to be much lower than that in trophoblast cells prepared from third-trimester placental tissues. Furthermore, expression of both tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 in cytotrophoblasts were significantly up-regulated in third-trimester when compared with the first-trimester of pregnancy. To further investigate the factors that caused the change of invasion-associated genes expression in trophoblast cells, we found that interleukin-10 (IL-10) could decrease the content of MMP-9 mRNA in cytotrophoblasts of first-trimester gestation, and the magnitude of suppression increased with increasing IL-10 concentration. The gradually reduced trophoblast invasion with gestational weeks might be on account of the change of proteolytic enzymes/activator/inhibitor genes expression. IL-10 could be one of the factors participating in the regulation of trophoblast invasion during gestational process.


A Mobile Phone App for the Self-Management of Pediatric Concussion: Development and Usability Testing.
Concussion is a common injury among Canadian children and adolescents that leads to a range of neurobehavioral deficits. However, noticeable gaps continue to exist in the management of pediatric concussion, with poor health outcomes associated with the inadequate application of best practice guidelines. The aim of this study was to describe the development and assess the usability of a mobile phone app to aid youth in the self-management of concussion. A secondary objective was to assess the usefulness of the app. An agile user-centered design approach was used to develop the technology, followed by a formative lab-based usability study for assessment and improvement proposals. Youths aged 10 to 18 years with a history of concussion and health care professionals involved in concussion management were recruited. This study included participants performing 12 tasks with the mobile phone app while using the think aloud protocol and the administration of the System Usability Scale (SUS), posttest questionnaire, and a semistructured interview. A mobile phone app prototype called NeuroCare, an easily accessible pediatric concussion management intervention that provides easy access to expert-informed concussion management strategies and helps guide youth in self-managing and tracking their concussion recovery, was developed. A total of 7 youths aged between 10 and 18 years with a history of concussion and 7 health care professionals were recruited. The mean SUS score was 81.9, mean task success rates were greater than 90% for 92% (11/12) of the tasks, 92% (11/12) of tasks had a total error frequency of less than 11 errors, and mean task completion times were less than 2 min for 100% of the tasks. Results suggest that participants rated this app as highly usable, acceptable to users, and that it may be useful in helping youth self-manage concussion.


Effect of decompressive craniectomy in the postoperative expansion of traumatic intracerebral hemorrhage: a propensity score-based analysis.
Traumatic intracerebral hemorrhage (TICH) represents approximately 13%-48% of the lesions after a traumatic brain injury (TBI), and hemorrhagic progression (HP) occurs in 38%-63% of cases. In previous studies, decompressive craniectomy (DC) has been characterized as a risk factor in the HP of TICH; however, few studies have focused exclusively on this relationship. The object of the present study was to analyze the relationship between DC and the growth of TICH and to reveal any correlation with the size of the craniectomy, degree of cerebral parenchymal herniation (CPH), or volumetric expansion of the TICH. The authors retrospectively analyzed the records of 497 adult patients who had been consecutively admitted after suffering a severe or moderate closed TBI. An inclusion criterion was presentation with one or more TICHs on the initial or control CT. Demographic, clinical, radiological, and treatment variables were assessed for associations. Two hundred three patients presenting with 401 individual TICHs met the selection criteria. TICH growth was observed in 281 cases (70.1%). Eighty-two cases (20.4%) underwent craniectomy without TICH evacuation. In the craniectomy group, HP was observed in 71 cases (86.6%); in the noncraniectomy group (319 cases), HP occurred in 210 cases (65.8%). The difference in the incidence of HP between the two groups was statistically significant (OR 3.41, p < 0.01). The mean area of the craniectomy was 104.94 ± 27.5 cm2, and the mean CPH distance through the craniectomy was 17.85 ± 11.1 mm. The mean increase in the TICH volume was greater in the groups with a craniectomy area > 115 cm2 and CPH > 25 mm (16.12 and 14.47 cm3, respectively, p = 0.01 and 0.02). After calculating the propensity score (PS), the authors followed three statistical methods-matching, stratification, and inverse probability treatment weighting (IPTW)-thereby obtaining an adequate balance of the covariates. A statistically significant relationship was found between HP and craniectomy (OR 2.77, p = 0.004). This correlation was confirmed with the three methodologies based on the PS with odds greater than 2. DC is a risk factor for the growth of TICH, and there is also an association between the size of the DC and the magnitude of the volume increase in the TICH.


Functional neural circuits that underlie developmental stuttering.
The aim of this study was to identify differences in functional and effective brain connectivity between persons who stutter (PWS) and typically developing (TD) fluent speakers, and to assess whether those differences can serve as biomarkers to distinguish PWS from TD controls. We acquired resting-state functional magnetic resonance imaging data in 44 PWS and 50 TD controls. We then used Independent Component Analysis (ICA) together with Hierarchical Partner Matching (HPM) to identify networks of robust, functionally connected brain regions that were highly reproducible across participants, and we assessed whether connectivity differed significantly across diagnostic groups. We then used Granger Causality (GC) to study the causal interactions (effective connectivity) between the regions that ICA and HPM identified. Finally, we used a kernel support vector machine to assess how well these measures of functional connectivity and granger causality discriminate PWS from TD controls. Functional connectivity was stronger in PWS compared with TD controls in the supplementary motor area (SMA) and primary motor cortices, but weaker in inferior frontal cortex (IFG, Broca's area), caudate, putamen, and thalamus. Additionally, causal influences were significantly weaker in PWS from the IFG to SMA, and from the basal ganglia to IFG through the thalamus, compared to TD controls. ICA and GC indices together yielded an accuracy of 92.7% in classifying PWS from TD controls. Our findings suggest the presence of dysfunctional circuits that support speech planning and timing cues for the initiation and execution of motor sequences in PWS. Our high accuracy of classification further suggests that these aberrant brain features may serve as robust biomarkers for PWS.


Nursing and the New Deal: we met the challenge.
The years of the great depression were marked with unemployment and economic ruin for many people. Americans were left feeling helpless and hopeless. After the 1932 presidential election of Franklin Roosevelt, his administration embarked on a course of government known as the New Deal. Many new and innovative programs were established to create jobs and a sense of hope for the public. This article will examine four programs that were of particular interest to nursing: the Federal Emergency Relief Act, the Civil Works Act, the Works Progress Act, and the Social Security Act. Nurses of the time embraced these programs. They participated in their development and implementation and made a difference in the lives of many desperate Americans.


[Prenatal care in Quixadá-CE according to SISPRENATAL's process indicators].
This is a descriptive, documental survey aimed at analyzing the process indicators of the Prenatal Information System (Sisprenatal) in the municipality of Quixadá, State of Ceará. The records of 1544 pregnant women from 2001 through 2004 were studied. Data were collected from February through April, 2005 in Sisprenatal, located in the Epidemiology sector of the local Health Secretary. The results were presented in a single chart containing absolute and relative frequency. They were analyzed in the light of current literature and the authors' experience. In the period was verified a growing percentage of pregnant women with indicators of at least six appointments, all the basic exams, anti-HIV test, antitetanic immunization and puerperal consultation, from zero percent (2001), to 2.6% (2002), to 5.68% (2003) to 21.11% (2004). Thus it is necessary to improve the use of the system, as well as the systematic reading of the process indicators, in order to obtain subsidies for the enhancement of the quality of prenatal assistance.


H19 and Igf2 monoallelic expression is regulated in two distinct ways by a shared cis acting regulatory region upstream of H19.
H19 and Igf2 are expressed in a monoallelic fashion from the maternal and paternal chromosomes, respectively. A region upstream of H19 has been shown to regulate such imprinted expression of both genes in cis. We have taken advantage of a loxP/cre recombinase-based strategy to delete this region in mice in a conditional manner to determine the temporal requirement of the upstream region in initiating and maintaining the imprinted expression of H19 and Igf2. Analysis of allele-specific expression of H19 and Igf2 and DNA methylation at the H19 promoter demonstrates that this region controls the monoallelic expression of the two genes in different ways, suggesting that it harbors two functionally distinct regulatory elements. Continued presence of the region is required to silence maternal Igf2 in accordance with its proposed role as an insulator. However, it does not have a direct role in keeping the paternal H19 promoter silenced. Instead, on the paternal chromosome, the upstream element mediates epigenetic modifications of the H19 promoter region during development, leading to transcriptional silencing of H19. Thereafter, its presence is redundant for preventing transcription. Presently, this temporal requirement of the silencing element appears to be a unique cis activity in the mammalian system. However, it is likely that other cis-acting elements, positive and negative, have the ability to effect stable changes in the chromatin structure and are not constantly required to give signals to the transcriptional machinery.


Public reactions to obesity-related health campaigns: a randomized controlled trial.
Despite numerous obesity-related health campaigns throughout the U.S., public perceptions of these campaigns have not been formally assessed. In addition, several recent publicized campaigns have come under criticism in the popular media for reinforcing stigmatization of obese people. Thus, research in this area is warranted. To systematically assess public perceptions of recent obesity-related public health campaigns in the U.S. RCT. The data were collected online in summer 2012 from a nationally representative sample of American adults (N=1085). Participants were randomly assigned to view 10 obesity-related health campaigns that were pretested and publicly criticized as being stigmatizing of obese people, or 10 campaigns that contained more-neutral content. Participants provided evaluations of each of the campaigns regarding the extent to which campaigns were rated to be stigmatizing of obese people, motivating for improving lifestyle behaviors, and promoting of self-efficacy for healthy behavior change. Participants additionally evaluated the appropriateness of the visual content depicted in each campaign. Analysis was completed in 2012. Stigmatizing campaigns were no more likely to instill motivation for improving lifestyle behaviors among participants than campaigns that were more neutral (OR=1.095, 95% CI=0.736, 1.630). Stigmatizing campaigns were also rated as inducing less self-efficacy (adjusted mean difference = -0.171 SD, 95% CI= -0.266, -0.076) and having less-appropriate visual content compared to less stigmatizing campaigns (adjusted difference in probability = -0.092, 95% CI= -0.124, -0.059). These findings remained consistent regardless of participants' body weight, and were generally consistent across sociodemographic predictors. This study highlights the need for careful selection of language and visual content used in obesity-related health campaigns, and provides support for efforts to portray obese people in a nonstigmatizing manner.


Oxygen and hydrogen stable isotope ratios of bulk needles reveal the geographic origin of Norway spruce in the European Alps.
Tracking timber is necessary in order to prevent illegal logging and protect local timber production, but there is as yet no suitable analytical traceability method. Stable isotope ratios in plants are known to reflect geographical variations. In this study we analysed four stable isotope ratios in order to develop a model able to identify the geographic origin of Norway spruce in the European Alps. δ18O, δ2H, δ13C and δ15N were measured in bulk needles of Picea abies sampled in 20 sites in and around the European Alps. Environmental and spatial variables were found to be related to the measured isotope ratios. An ordinary least squares regression was used to identify the most important factor in stable isotope variability in bulk needles. Spatial autocorrelation was tested for all isotope ratios by means of Moran's I. δ18O, δ2H and δ15N values differed significantly between sites. Distance from the coast had the greatest influence on δ2H, while latitude and longitude were strongly related to δ18O. δ13C values did not appear to have any relationship with geographical position, while δ15N values were influenced by distance from the motorway. The regression model improved the explanatory power of the spatial and environmental variables. Positive spatial autocorrelations were found for δ18O and δ2H values. The δ 18O, δ2H and δ15N values in P. abies bulk needles are a suitable proxy to identify geographic origin as they vary according to geographical position. Although the regression model showed the explanatory variables to have significant power and stability, we conclude that our model might be improved by multivariate spatial interpolation of the δ 18O and δ2H values.


Impact of metformin on malignancy in solid organ transplantation.
Malignancy after solid organ transplant is a common occurrence that is associated with increased morbidity and mortality. Literature in the general diabetic population has identified possible antineoplastic properties of metformin. This retrospective study aims to determine if metformin results in a malignancy risk reduction in a cohort of diabetic kidney, liver, and heart transplant recipients. The population included transplant recipients without use of metformin at any time point (DMO arm, n = 147) and those with use of metformin (DMM arm, n = 172); the two arms were matched based on organ type and transplant date prior to application of exclusion criteria. Recipients with prior malignancy, malignancy before diabetes diagnosis, and metformin duration <30 days were excluded. The primary endpoint of malignancy first occurrence post-transplant was not found to be statistically significant at 1, 5, 10, and 15 years. In the subgroup of heart transplant recipients, there was a significant reduction in malignancy at 15 years post-transplant. Older age and Caucasian race were identified as significant risk factors for malignancy, while never smoker was a protective factor. Metformin use in this solid organ transplant cohort was not found to significantly reduce malignancy risk compared to use of other anti-diabetic agents.


Pharmacogenetics of Lipid-lowering Therapies.
Cardiovascular disease is associated with nonmodifiable risk factors such as age, gender, and genetic background, and with modifiable risk factors such as lipid concentrations. Lowering serum lipid levels has been demonstrated to slow the progression of, or even induce regression in, atherosclerosis. However, like any other drug treatment, the magnitude of plasma lipid responses to drug therapies varies considerably among individuals. Pharmacogenetics provides the experimental basis to understand the variability in response to drugs as a function of the individual genetic makeup. Information from small clinical trials reveals that several candidate genes may hold some promise in our quest to predict individual success to hypolipemic drug treatment. However, the current clinical relevance of this knowledge is quite limited due to the small effects observed for each of the genetic markers examined. Future progress in this area will be driven by studying gene-gene and gene-treatment interactions in much larger patient populations.


Fibroblast growth factor 9 activates anti-oxidative functions of Nrf2 through ERK signalling in striatal cell models of Huntington's disease.
Huntington's disease (HD) is a heritable neurodegenerative disorder, and has been characterized as an increase of oxidative stress in brain regions. In our previous results, we showed fibroblast growth factor 9 (FGF9) provides neuroprotective functions to suppress cell death in HD striatal cells dominantly through ERK signalling. However, whether the working mechanism of FGF9 is related to anti-oxidative stress in HD is still unknown. In this study, STHdhQ7/Q7 (Q7) and STHdhQ111/Q111 (Q111) striatal knock-in cell lines were used to examine the neuroprotective effects of FGF9 against oxidative stress in HD. Results show that FGF9 alleviates oxidative stress induced by starvation in Q7 and Q111 cells. The treatment of FGF9 not only induces upregulation and activation of nuclear factor erythroid 2-like 2 (Nrf2), a critical transcription factor for anti-oxidative stress, but also further upregulates its downstream targets, such as superoxide dismutase 2, gamma-glutamylcysteine synthetase and glutathione reductase. Furthermore, blockage of the Nrf2 pathway abolishes the anti-oxidative functions of FGF9, and inhibition of ERK signalling reduces the activation of the FGF9-Nrf2 pathway, resulting in higher level of oxidative stress in HD cells. These results support the neuroprotective effects of FGF9 against oxidative stress through the ERK-Nrf2 pathway, and imply one of potential strategies for therapy of HD.


Genome Analysis of Melon Necrotic Spot Virus Incursions and Seed Interceptions in Australia.
Melon necrotic spot virus (MNSV) was detected in field-grown Cucumis melo (rockmelon) and Citrullus lanatus (watermelon) plants in the Sunraysia district of New South Wales and Victoria, Australia, in 2012, 2013, and 2016, and in two watermelon seed lots tested at the Australian border in 2016. High-throughput sequencing was used to generate near full-length genomes of six isolates detected during the incursions and seed testing. Phylogenetic analysis of the genomes suggests that there have been at least two incursions of MNSV into Australia and none of the field isolates were the same as the isolates detected in seeds. The analysis indicated that one watermelon field sample (L10), the Victorian rockmelon field sample, and two seed interception samples may have European origins. The results showed that two isolates (L8 and L9) from watermelon were divergent from the type MNSV strain (MNSV-GA, D12536.2) and had 99% nucleotide identity to two MNSV isolates from human stool collected in the United States (KY124135.1, KY124136.1). These isolates also had high nucleotide pairwise identity (96%) to a partial sequence from a Spanish MNSV isolate (KT962848.1). The analysis supported the identification of three previously described MNSV genotype groups: EU-LA, Japan melon, and Japan watermelon. To account for the greater diversity of hosts and geographic regions of the MNSV isolates used in this study, it is suggested that the genotype groups EU-LA, Japan melon, and Japan watermelon be renamed to groups I, II, and III, respectively. The divergent isolates L8 and L9 from this study and the stool isolates from the United States formed a fourth genotype group, group IV. Soil collected from the site of the Victorian rockmelon MNSV outbreak was found to contain viable MNSV and the virus vector, a chytrid fungus, Olpidium bornovanus (Sahtiyanci) Karling, 18 months after the initial MNSV detection. This is a first report of O. bornovanus from soil sampled from an MNSV-contaminated site in Australia.


Microbial growth on oxalate by a route not involving glyoxylate carboligase.
1. The metabolism of oxalate by the pink-pigmented organisms, Pseudomonas AM1, Pseudomonas AM2, Protaminobacter ruber and Pseudomonas extorquens has been compared with that of the non-pigmented Pseudomonas oxalaticus. 2. During growth on oxalate, all the organisms contain oxalyl-CoA decarboxylase, formate dehydrogenase and oxalyl-CoA reductase. This is consistent with oxidation of oxalate to carbon dioxide taking place via oxalyl-CoA, formyl-CoA and formate as intermediates, and also reduction of oxalate to glyoxylate taking place via oxalyl-CoA. 3. The pink-pigmented organisms, when grown on oxalate, contain l-serine-glyoxylate aminotransferase and hydroxypyruvate reductase but do not contain glyoxylate carboligase. The converse of this obtains in oxalate-grown Ps. oxalaticus. This indicates that, in contrast with Ps. oxalaticus, synthesis of C(3) compounds from oxalate by the pink-pigmented organisms occurs by a variant of the ;serine pathway' used by Pseudomonas AM1 during growth on C(1) compounds. 4. Evidence in favour of this scheme is provided by the finding that a mutant of Pseudomonas AM1 that lacks hydroxypyruvate reductase is not able to grow on oxalate.


Association between clinically meaningful behavior problems and overweight in children.
To determine whether there is a relationship between clinically meaningful behavior problems and concurrent and future overweight in 8- to 11-year-old children. 1998 National Longitudinal Survey of Youth interview data for 8- to 11-year-old children and their mothers were analyzed. A Behavior Problems Index score >90th percentile was considered clinically meaningful. Child overweight was defined as a body mass index (BMI) >or=95th percentile for age and sex. Multiple logistic regression was used to control for potential confounders (selected a priori): child's sex, race, use of behavior-modifying medication, history of academic retention, and hours of television per day; maternal obesity, smoking status, marital status, education, and depressive symptoms; family poverty status; and Home Observation for Measurement of the Environment-Short Form (HOME-SF) cognitive stimulation score. In an attempt to elucidate temporal sequence, a second analysis was conducted with a subsample of normal-weight children who became overweight between 1996 and 1998 while controlling for BMI z score in 1996. The sample included 755 mother-child pairs. Of the potential confounding variables, race, maternal obesity, academic grade retention, maternal education, poverty status, and HOME-SF cognitive stimulation score acted as joint confounders, altering the relationship between behavior problems and overweight in the multiple logistic regression model. With these covariates in the final model, behavior problems were independently associated with concurrent child overweight (adjusted odds ratio: 2.95; 95% confidence interval: 1.34-6.49). The relationship was strengthened in the subsample of previously normal-weight children, with race, maternal obesity, HOME-SF cognitive stimulation score, and 1996 BMI z score acting as confounders (adjusted odds ratio: 5.23; 95% confidence interval: 1.37-19.9). Clinically meaningful behavior problems in 8- to 11-year-old children were independently associated with an increased risk of concurrent overweight and becoming overweight in previously normal-weight children.


Genetic parameters for atypical reproductive patterns in dairy cows estimated from in-line milk progesterone profiles.
Our aim was to estimate genetic parameters of atypical reproductive patterns and estimate their genetic correlation with milk production and classical fertility traits for commercial dairy cows. In contrast with classical fertility traits, atypical reproductive patterns based on in-line milk progesterone profiles might have higher heritability and lower genetic correlation with milk production. We had in-line milk progesterone profiles available for 12,046 cycles in 4,170 lactations of 2,589 primiparous and multiparous cows (mainly Holstein Friesian) from 14 herds. Based on progesterone profiles, 5 types of atypical reproductive patterns in a lactation were defined: delayed ovulation types I and II, persistent corpus luteum types I and II, and late embryo mortality. These atypical patterns were detected in 14% (persistent corpus luteum type II) to 21% (persistent corpus luteum type I) of lactations. In 47% of lactations, at least 1 atypical pattern was detected. Threshold model heritabilities for atypical reproduction patterns ranged between 0.03 and 0.14 and for most traits were slightly higher compared with classical fertility traits. The genetic correlation between milk yield and calving interval was 0.56, whereas genetic correlations between milk yield and atypical reproductive patterns ranged between -0.02 and 0.33. Although most of these correlations between milk yield and atypical reproductive patterns are still unfavorable, they are lower compared with the correlations between classical fertility traits and milk yield. Therefore selection against atypical reproductive patterns may relax some constraints in current dairy breeding programs, to enhance genetic progress in both fertility and milk yield at a steady pace. However, as long as the target trait for fertility is calving interval, atypical reproductive patterns will not add additional value to the breeding goal in the near future due to the low number of available records.


Image coding using variable-rate side-match finite-state vector quantization.
Future B-ISDN (broadband integrated services digital network) users will be able to send various kinds of information, such as voice, data, and image, over the same network and send information only when necessary. It has been recognized that variable-rate encoding techniques are more suitable than fixed-rate techniques for encoding images in a B-ISDN environment. A new variable-rate side-match finite-state vector quantization with a block classifier (CSMVQ) algorithm is described. In an ordinary fixed-rate SMVQ, the size of the state codebook is fixed. In the CSMVQ algorithm presented, the size of the state codebook is changed according to the characteristics of the current vector which can be predicted by a block classifier. In experiments, the improvement over SMVQ was up to 1.761 dB at a lower bit rate. Moreover, the improvement over VQ can be up to 3 dB at nearly the same bit rate.


Beta-cell MiRNA-503-5p Induced by Hypomethylation and Inflammation Promotes Insulin Resistance and β-Cell Decompensation.
Chronic inflammation promotes pancreatic β-cell decompensation to insulin resistance due to local accumulation of supraphysiologic IL-1β levels. However, the underlying molecular mechanism(s) remains elusive. We show that miR-503-5p is exclusively upregulated in islets from type 2 diabetic humans and rodents due to its promoter hypomethylation and increased local IL-1β levels. Beta-cell-specific miR-503 transgenic (βTG) mice display mild or severe diabetes in a time- and expressiondependent manner. By contrast, deletion of the miR-503 cluster protects mice from high-fat-diet-induced insulin resistance and glucose intolerance. Mechanistically, miR-503-5p represses JNK interacting protein 2 (JIP2) translation to activate mitogen-activated protein kinases (MAPKs) signaling cascades, thus inhibiting glucose-stimulated insulin secretion (GSIS) and compensatory β-cell proliferation. In addition, β-cell miR-503-5p is packaged in nano-vesicles to dampen insulin signaling transduction in liver and adipose tissues by targeting insulin receptors (INSR). Notably, specifically blocking the miR-503 cluster in β cells effectively remits ageing-associated diabetes through recovery of GSIS capacity and insulin sensitivity. Our finding demonstrated that β-cell miR-503-5p is required for the development of insulin resistance and β-cell decompensation, providing a potential therapeutic target against diabetes.


From Topological Analyses to Functional Modeling: The Case of Hippocampus.
Topological data analyses are widely used for describing and conceptualizing large volumes of neurobiological data, e.g., for quantifying spiking outputs of large neuronal ensembles and thus understanding the functions of the corresponding networks. Below we discuss an approach in which convergent topological analyses produce insights into how information may be processed in mammalian hippocampus-a brain part that plays a key role in learning and memory. The resulting functional model provides a unifying framework for integrating spiking data at different timescales and following the course of spatial learning at different levels of spatiotemporal granularity. This approach allows accounting for contributions from various physiological phenomena into spatial cognition-the neuronal spiking statistics, the effects of spiking synchronization by different brain waves, the roles played by synaptic efficacies and so forth. In particular, it is possible to demonstrate that networks with plastic and transient synaptic architectures can encode stable cognitive maps, revealing the characteristic timescales of memory processing.


The effect of freezing on human cortical cataracts.
To ascertain a direct relationship between the light scattering intensity and the particle scattering the light, thin (6-20 micrometers) sections were cut from frozen human cataracts. Freezing for 15 min (-10 C) and then thawing had no effect on the clarity or light scattering properties of nuclear cataracts. Cortical cataracts treated in the same manner appeared to be much clearer than they were prior to freeze-thawing. Light scattering measurements were obtained as a function of scattering angle, both in the I parallel and I perpendicular mode; a He-Ne laser was used. Results showed a reduction in the light scattering intensity in the I parallel and I perpendicular modes with repeated cyclical freezing and thawing and that this decrease was most pronounced at higher scattering angles. Morphologic data showed that much of the anatomical heterogeneity present in the human cortical and rat lens osmotic cataract disappeared upon freeze-thawing. However, even though the anatomical heterogeneity was less, there was persistence of serious cellular anatomical disorganization in grossly transparent parts of the cortex. This suggests that transparency is not dependent upon the presence of intact fiber cells. The importance of these findings to studies of light scattering, cataract classification and analysis of cryoextracted human lenses is discussed.


Ocular side effects of target therapy and immunotherapy in patients with cutaneous malignant melanoma.
To examine the nature and frequency of ocular side effects due to systemic target therapy with BRAF and MEK inhibitors as well as immunotherapy with cytotoxic T-lymphocyte antigen 4 (CTLA-4) and programmed cell death 1 (PD-1) monoclonal antibodies used in the treatment of cutaneous malignant melanoma (CMM). While proven effective in cancer treatment, target therapy and immunotherapy have been associated with ocular side effects likely due to their ability to alter the immune privilege of the eye. We conducted a retrospective chart review of patients undergoing target and immunotherapy for CMM and documented all associated eye findings. We reviewed the records of 34 patients receiving target and immunotherapy for CMM who were examined in the academic ophthalmology clinic between 2012 and 2017. Ocular side effects were present in 41.1% of patients in this study with 14.7% presenting with uveitis. Patients undergoing therapy with either vemurafenib only or dabrafenib/trametinib combination therapies comprised 70.5% of the study cohort. Ocular side effects occurred in 45.5% and 46.1% of patients on vemurafenib and dabrafenib/trametinib combination therapy, respectively. About 47.5% of males presented with ocular side effects compared to 30.5% of females. Notably, 13/14 patients with ocular symptoms recovered. This study highlights the frequency of ocular side effects in patients treated with target therapy and immunotherapy for CMM and shows that symptom resolution can be effectively achieved with proper ophthalmic care. Further research is required to answer whether cessation of these therapies is mandatory during ophthalmic treatment.


Omega-3 fatty acids and incident type 2 diabetes: the Singapore Chinese Health Study.
The role of omega-3 (n-3) fatty acids (FAs) in the development of type 2 diabetes is uncertain, especially with regard to any differential influence of α-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). The objective was to examine the association between total omega-3 FAs, marine omega-3 (EPA, DHA), nonmarine omega-3 (ALA), and omega-6 (n-6) FAs and omega-6:omega-3 ratio and risk of type 2 diabetes in a Chinese population in Singapore. The analysis included 43,176 Chinese men and women free of chronic disease, aged 45-74 y, in the Singapore Chinese Health Study. Baseline data collection occurred between 1993 and 1998, with follow-up interviews between 1999 and 2004. Cox regression models were used to examine the associations between FA intakes at baseline and risk of developing diabetes. Increased intakes of total omega-3 FAs were inversely associated with diabetes incidence [hazard ratio (HR) for the fifth compared with the first quintile: 0.78; 95% CI: 0.65, 0.94; P for trend = 0.02]. Omega-3 FAs from marine sources were not associated with diabetes risk, whereas nonmarine omega-3 FA intake was strongly associated (HR for the fifth compared with the first quintile: 0.79; 95% CI: 0.67, 0.93; P for trend = 0.004). Omega-6 and omega-6:omega-3 ratio were not associated with incidence of type 2 diabetes. Consumption of nonmarine sources (ALA) of omega-3 FAs is associated with a decreased risk of type 2 diabetes in Chinese Singaporeans.


Metformin and AMP Kinase Activation Increase Expression of the Sterol Transporters ABCG5/8 (ATP-Binding Cassette Transporter G5/G8) With Potential Antiatherogenic Consequences.
The mechanisms underlying the cardiovascular benefit of the anti-diabetic drug metformin are poorly understood. Recent studies have suggested metformin may upregulate macrophage reverse cholesterol transport. The final steps of reverse cholesterol transport are mediated by the sterol transporters, ABCG5 (ATP-binding cassette transporter G5) and ABCG8 (ATP-binding cassette transporter G8), which facilitate hepato-biliary transport of cholesterol. This study was undertaken to assess the possibility that metformin induces Abcg5 and Abcg8 expression in liver and to elucidate the underlying mechanisms. Metformin-treated mouse or human primary hepatocytes showed increased expression of Abcg5/8 and the bile salt export pump, Bsep. Administration of metformin to Western-type diet-fed mice showed significant upregulation of Abcg5/8 and Bsep. This resulted in increased initial clearance of 3H-cholesteryl ester HDL (high-density lipoprotein) from plasma. However, fecal 3H-cholesterol output was only marginally increased, possibly reflecting increased hepatic Ldlr (low-density lipoprotein receptor) expression, which would increase nonradiolabeled cholesterol uptake. Abcg5/8 undergo strong circadian variation. Available chromatin immunoprecipitation-Seq data suggested multiple binding sites for Period 2, a transcriptional repressor, within the Abcg5/8 locus. Addition of AMPK (5' adenosine monophosphate-activated protein kinase) agonists decreased Period 2 occupancy, suggesting derepression of Abcg5/8. Inhibition of ATP citrate lyase, which generates acetyl-CoA from citrate, also decreased Period 2 occupancy, with concomitant upregulation of Abcg5/8. This suggests a mechanistic link between feeding-induced acetyl-CoA production and decreased cholesterol excretion via Period 2, resulting in inhibition of Abcg5/8 expression. Our findings provide partial support for the concept that metformin may provide cardiovascular benefit via increased reverse cholesterol transport but also indicate increased Ldlr expression as a potential additional mechanism. AMPK activation or ATP citrate lyase inhibition may mediate antiatherogenic effects through increased ABCG5/8 expression.


Vestibular migraine.
Vestibular migraine is now considered a distinct diagnostic entity by both the Barany Society and the International Headache Society. The recognition of vestibular migraine as a diagnostic entity required decades and was presaged by several reports indicating that a large proportion of patients with migraine headaches have vestibular symptoms and that a large proportion of patients with undiagnosed episodic vestibular symptoms have migraine headache. Despite the availability of diagnostic criteria for vestibular migraine, challenges to diagnosis include variability in terms of the character of dizziness, the presence or absence of clearly defined attacks, the duration of attacks, and the temporal association between headache or other migrainous features and vestibular symptoms. Also, symptoms of vestibular migraine often overlap with symptoms of other causes of dizziness, especially Ménière's disease and benign paroxysmal positional vertigo (BPPV). This article will discuss the demographics, epidemiology, clinical manifestations, physical examination findings, laboratory testing, comorbidities, treatment options, and pathophysiology of vestibular migraine. Future research in the field of vestibular migraine should include both clinical and basic science efforts to better understand the pathophysiology of this condition. Controlled treatment trials for vestibular migraine are desperately needed.


Neonatal risks from in vitro fertilization and delayed motherhood.
Delayed childbearing (DC) is common in most Western countries. The average age of first-time mothers increased in United States from 21.4 years in 1970 to 25.0 years in 2006 and from 25.4 to 30.8 years in Australia in the same period. It is commonly believed that this has no ominous consequences. But several negative consequences of this behavior are described: stillbirth, prematurity, twins, birth anomalies. Age also decreases women's fertility, thus many couples undergo in vitro fertilization. And we highlight a paradox: medical reproduction techniques decreases their effectiveness with maternal age, but their availability can be an incentive to postpone parenthood. Of course the risks of delayed parenthood involve a minority of cases, but are parents entitled to accept any risk on the behalf of their baby A complete information would make people cautious before deciding to postpone childbearing, though this is often an obliged rather that a free choice: the consumerist society pressure and the difficulty to find an employment have their heavy weight in this choice. But if this choice is not really free, people's interest is to overcome these pressures and to claim for a real broad choice on when becoming parent, despite the pressures made by their cultural environment to postpone parenthood. Moreover, even reproductive techniques have some risks. Unfortunately, mass media often praise and endorse DC, disregarding the increase of premature babies born because of DC, a real alarm for public health. Pediatricians should discourage the culture that makes DC a normal event.


How can healthcare organizations implement patient-centered care? Examining a large-scale cultural transformation.
Healthcare organizations increasingly are focused on providing care which is patient-centered rather than disease-focused. Yet little is known about how best to transform the culture of care in these organizations. We sought to understand key organizational factors for implementing patient-centered care cultural transformation through an examination of efforts in the US Department of Veterans Affairs. We conducted multi-day site visits at four US Department of Veterans Affairs medical centers designated as leaders in providing patient-centered care. We conducted qualitative semi-structured interviews with 108 employees (22 senior leaders, 42 middle managers, 37 front-line providers and 7 staff). Transcripts of audio recordings were analyzed using a priori codes based on the Consolidated Framework for Implementation Research. We used constant comparison analysis to synthesize codes into meaningful domains. Sites described actions taken to foster patient-centered care in seven domains: 1) leadership; 2) patient and family engagement; 3) staff engagement; 4) focus on innovations; 5) alignment of staff roles and priorities; 6) organizational structures and processes; 7) environment of care. Within each domain, we identified multi-faceted strategies for implementing change. These included efforts by all levels of organizational leaders who modeled patient-centered care in their interactions and fostered willingness to try novel approaches to care amongst staff. Alignment and integration of patient centered care within the organization, particularly surrounding roles, priorities and bureaucratic rules, remained major challenges. Transforming healthcare systems to focus on patient-centered care and better serve the "whole" patient is a complex endeavor. Efforts to transform healthcare culture require robust, multi-pronged efforts at all levels of the organization; leadership is only the beginning. Challenges remain for incorporating patient-centered approaches in the context of competing priorities and regulations. Through actions within each of the domains, organizations may begin to truly transform to patient-driven care.


Discovery of shared genomic loci using the conditional false discovery rate approach.
In recent years, genome-wide association study (GWAS) sample sizes have become larger, the statistical power has improved and thousands of trait-associated variants have been uncovered, offering new insights into the genetic etiology of complex human traits and disorders. However, a large fraction of the polygenic architecture underlying most complex phenotypes still remains undetected. We here review the conditional false discovery rate (condFDR) method, a model-free strategy for analysis of GWAS summary data, which has improved yield of existing GWAS and provided novel findings of genetic overlap between a wide range of complex human phenotypes, including psychiatric, cardiovascular, and neurological disorders, as well as psychological and cognitive traits. The condFDR method was inspired by Empirical Bayes approaches and leverages auxiliary genetic information to improve statistical power for discovery of single-nucleotide polymorphisms (SNPs). The cross-trait condFDR strategy analyses separate GWAS data, and leverages overlapping SNP associations, i.e., cross-trait enrichment, to increase discovery of trait-associated SNPs. The extension of the condFDR approach to conjunctional FDR (conjFDR) identifies shared genomic loci between two phenotypes. The conjFDR approach allows for detection of shared genomic associations irrespective of the genetic correlation between the phenotypes, often revealing a mixture of antagonistic and agonistic directional effects among the shared loci. This review provides a methodological comparison between condFDR and other relevant cross-trait analytical tools and demonstrates how condFDR analysis may provide novel insights into the genetic relationship between complex phenotypes.


Endothelial cell response to chemical, biological, and physical cues in bioactive hydrogels.
The highly tunable biological, chemical, and physical properties of bioactive hydrogels enable their use in an array of tissue engineering and drug delivery applications. Systematic modulation of these properties can be used to elucidate key cell-material interactions to improve therapeutic effects. For example, the rate and extent of endothelialization are critical to the long-term success of many blood-contacting devices. To this end, we have developed a bioactive hydrogel that could be used as coating on cardiovascular devices to enhance endothelial cell (EC) adhesion and migration. The current work investigates the relative impact of hydrogel variables on key endothelialization processes. The bioactive hydrogel is based on poly(ethylene glycol) (PEG) and a streptococcal collagen-like (Scl2-2) protein that has been modified with integrin α1β1 and α2β1 binding sites. The use of PEG hydrogels allows for incorporation of specific bioactive cues and independent manipulation of scaffold properties. The selective integrin binding of Scl2-2 was compared to more traditional collagen-modified PEG hydrogels to determine the effect of integrin binding on cell behavior. Protein functionalization density, protein concentration, and substrate modulus were independently tuned with both Scl2-2 and collagen to determine the effect of each variable on EC adhesion, spreading, and migration. The findings here demonstrate that increasing substrate modulus, decreasing functionalization density, and increasing protein concentration can be utilized to increase EC adhesion and migration. Additionally, PEG-Scl2-2 hydrogels had higher migration speeds and proliferation over 1 week compared with PEG-collagen gels, demonstrating that selective integrin binding can be used to enhance cell-material interactions. Overall, these studies contribute to the understanding of the effects of matrix cues on EC interactions and demonstrate the strong potential of PEG-Scl2-2 hydrogels to promote endothelialization of blood-contacting devices.


Standardization of doctoral study in agricultural and extension education: is the field of study mature enough for achievement of the optimum degree of order?
Agricultural and extension education--or some derivative name--is a field of study leading to the doctoral degree in universities around the world. Is there are body of knowledge or a taxonomy of the knowledge--e.g., a knowledge domain--that one should possess with a doctorate in agricultural and extension education? The purpose of this paper was to synthesize the work of researchers who attempted to define the field of study, with a taxonomy comprising the knowledge domains (standards) and knowledge objects--structured interrelated sets of data, knowledge, and wisdom--of the field of study. Doctoral study in agricultural and extension education needs a document that provides for rules and guidelines--rules and guidelines that in turn provide for common and repeated use--all leading to achievement of an optimum degree of order in the context of academic, scholarly, and professional practice in agricultural and extension education. Thus, one would know in broad categories the knowledge, skills, and abilities possessed by one who holds a doctoral degree in agricultural and extension education. That is, there would exist a standard for doctoral degrees in agricultural and extension education. A content analysis of three previous attempts to categorize knowledge in agricultural and extension education served as the primary technique to create a new taxonomy--or to confirm an existing taxonomy--for doctoral study in agricultural and extension education. The following coalesced as nine essential knowledge domains for a doctorate in agricultural and extension education: (1) history, philosophy, ethics, and policy; (2) agricultural/rural development; (3) organizational development and change management; (4) planning, needs assessment, and evaluation; (5) learning theory; (6) curriculum development and instructional design; (7) teaching methods and delivery strategies; (8) research methods and tools; and, (9) scholarship and communications.


Immediate postoperative CT to assess the correct positioning of a ventricular catheter.
Positioning of the ventricular catheter is safe using standard techniques, but in some cases there may be failure of a shunt because of incorrect placement. We carried out CT at the end of the surgical operation on 10 patients who had undergone cerebrospinal fluid shunting while still anaesthetized. In three cases a malpositioned catheter was detected. Immediate reoperation allowed readjustment of the ventricular catheter. Early postoperative CT appears to be useful for verifying correct placement.


Calcium control of ciliary arrest in mussel gill cells.
After several hours in 20 mM sodium phosphate and 40 mM KCI (pH 7.4) or similar simple solutions, ciliated cells exfoliate en masse from stripped gill epithelium of freshwater mussels, e.g., Elliptio complanatus. Three types of ciliated cells--lateral (L), laterofrontal (LF), and frontal (F)--can be distiniguished and counted separately in the suspensions. About one-half of the cells of each type remain motile. Motility is unaffected by addition of 10(-5) M A23187 or 10(-2) M Ca+2 added separately, but when ionophore and Ca+2 are added together, ciliary beat is largely arrested. Treatment of the cells with Triton X-100 (Rohm & Haas Co., Philadelphia, Pa.) results in a total loss of motility as the ciliary membrane becomes disrupted. Such models can be reactivated by addition of ATP and Mg+2. All ciliated cell types are reactivated to about the same extent. At least 80% of the activity of the untreated preparation returns. Ca+2-EGTA buffers added to the reactivating solutions permit titration of free Ca+2 concentration vs. percent motility. Activity is unchanged for all cell types at Ca+2 less than 10(-7) M; at 10(-6) Ca+2, L cilia of all cell types are arrested differentially, whereas at Ca+2 greater than 10(-4) M most cilia of all cell types are arrested. We conclude: (a) that increasing cytoplasmic Ca+2 is directly responsible for ciliary arrest, (b) that the readily reversible physiological arrest response of the L cilia in the intact gill is caused by a rise in free Ca+2 in narrow limits from ca. 5 x 10(-7) M to ca. 8 x 10(-7) M, and (c) that the site which is sensitive to Ca+2 is part of the ciliary axoneme or the basal apparatus.


Mothers' Knowledge, Health Beliefs and Intentions to Vaccinate Their Daughters against Human Papillomavirus (HPV) in Korea.
Abstract This study aimed to identify the factors influencing Korean mothers' intentions to vaccinate their daughters against human papillomavirus (HPV). In Korea, a mother's will is a key determinant for daughters receiving the cervical cancer vaccine. The factors influencing mothers' intentions to vaccinate their daughters against HPV were the perceived benefits of the vaccine, having acquaintances with cervical cancer experiences, awareness of the cervical cancer vaccine, believing that one's daughter needs regular Pap smears, and occupation; these variables explained 32% of the variance. The perceived benefits of the vaccine were the strongest predictor of mothers' intentions. The findings of the current study suggested that mothers' perceived benefits about the HPV vaccine play an important role in their choice to vaccinate their daughters.


CCBuilder: an interactive web-based tool for building, designing and assessing coiled-coil protein assemblies.
The ability to accurately model protein structures at the atomistic level underpins efforts to understand protein folding, to engineer natural proteins predictably and to design proteins de novo. Homology-based methods are well established and produce impressive results. However, these are limited to structures presented by and resolved for natural proteins. Addressing this problem more widely and deriving truly ab initio models requires mathematical descriptions for protein folds; the means to decorate these with natural, engineered or de novo sequences; and methods to score the resulting models. We present CCBuilder, a web-based application that tackles the problem for a defined but large class of protein structure, the α-helical coiled coils. CCBuilder generates coiled-coil backbones, builds side chains onto these frameworks and provides a range of metrics to measure the quality of the models. Its straightforward graphical user interface provides broad functionality that allows users to build and assess models, in which helix geometry, coiled-coil architecture and topology and protein sequence can be varied rapidly. We demonstrate the utility of CCBuilder by assembling models for 653 coiled-coil structures from the PDB, which cover >96% of the known coiled-coil types, and by generating models for rarer and de novo coiled-coil structures. CCBuilder is freely available, without registration, at http://coiledcoils.chm.bris.ac.uk/app/cc_builder/.


Sweetpotato seed exchange systems and knowledge on sweetpotato viral diseases among local farmers in Acholi Sub Region-Northern Uganda.
Viral infections greatly limit sweetpotato yields. Good farming practices are critical for effective disease management. 383 Ugandan farmers were surveyed to document symptom incidence, crop-management practices, and buyer preferences. Results showed that 89.27% farmers grow sweet potatoes yearly and 62.76% of these farmers were female. A total of 56.83% farmers obtained vine seeds from their previous gardens, 25.85% from neighbours, and 12.20% purchased. Only ~8% of sellers and ~4% of buyers were selected for disease-free materials. None of the farmers who used vine-cutting knives sterilised them. Almost half of farmers (47%) observed whitefly or aphids but most were unaware they are viral vectors. Most farmers (77%) observed viral symptoms, but few (<2%) recognised them as infections. Insufficient knowledge of sweetpotato viruses and their vectors is common and increases the risk of spread. Practices like vine selling, sharing of vines coupled with insufficient knowledge on sweet potato viruses and its vectors among farmers increase the risk of virus spread among different farms.


Relationship between a gum-chewing routine and oral, physical, and cognitive functions of community-dwelling older adults: A Kashiwa cohort study.
As associations between oral function and general health have been reported in community-dwelling older adults, easily implementable preventive measures are urgently required. We focused on the health benefits of gum chewing, as no studies have been carried out on the impact of gum-chewing routines on the health of older adults. This cross-sectional study aimed to determine whether the gum-chewing routine is associated with oral, physical and cognitive functions in community-dwelling older adults. This study included 1617 community-dwelling older participants in a health survey carried out in 2021. The gum-chewing routine and weekly chewing time were assessed using a self-administered questionnaire. The outcome measures, including actual measurements of oral function, physical function, cognitive function, dietary intake and lifestyle, were evaluated using self-administered questionnaires or health surveys. We analyzed 1474 (mean age 76.1 ± 5.8 years, 45% women) participants for whom all data were not missing, and 14% of them had a gum-chewing routine for more than 30 min weekly. Oral functions were significantly higher in older adults with a gum-chewing routine, and there were substantially fewer participants with oral frailty (adjusted odds ratio 0.581, 95% confidence interval 0.340-0.993). Additionally, cognitive and physical functions, including grip strength, were significantly higher in the gum-chewing routine group. Community-dwelling older adults with a gum-chewing routine have higher oral, physical and cognitive functions. These findings indicate that a gum-chewing routine might contribute to maintaining oral function and preventing frailty. Geriatr Gerontol Int 2023; ••: ••-••.


Multicomponent cognitive-behavioural intervention to improve sleep in adolescents: a multiple baseline design.
Adolescents are prone to sleep problems that have unique developmental aspects and contribute to physical, emotional, and behavioural problems. This study evaluated an individualized, multicomponent intervention that considered developmental factors, and promoted age-appropriate autonomy in three adolescent females with disrupted sleep. Adolescents recorded sleep data on daily logs. A nonconcurrent multiple baseline design was used to evaluate a cognitive-behavioural intervention including sleep hygiene training, bedtime routine development, cognitive restructuring, relaxation training, stimulus control, sleep restriction, bedtime fading, and problem-solving, along with clinically indicated individualization. Outcomes demonstrated clinically meaningful improvements and decreased variability in sleep parameters following intervention. Each participant's sleep log data indicated improvement in, or maintenance of, adequate total sleep time (TST), decreased sleep onset latency (SOL), improved sleep efficiency (SE), improvement in time of sleep onset, and decreased or continued low frequency of night awakenings (NA). Anecdotally, adolescents and parents reported improvement in daytime functioning, coping, and sense of wellbeing. These cases highlight the potential for cognitive-behavioural interventions to facilitate healthy sleep in adolescents with challenging sleep problems.


Enhanced production of human mini-proinsulin in fed-batch cultures at high cell density of Escherichia coli BL21(DE3)[pET-3aT2M2].
Synthesis of recombinant protein (human mini-proinsulin) is investigated in fed-batch cultures at high cell concentration of recombinant Escherichia coli BL21(DE3)[pET-3aT2M2]. Transcription of the recombinant gene is controlled by a T7 promoter system. The human mini-proinsulin is characterized by a C-chain peptide consisting of only nine amino acids, whereas the C-chain peptide of natural human proinsulin is made up of 35 amino acids. It is expressed in a fusion protein with a small fusion partner (a peptide with 18 amino acids) and finally aggregated into insoluble inclusion bodies in cytoplasm of recombinant E. coli. The fermentative production of this small fusion mini-proinsulin may be of great advantage in enhancing the yield of human insulin. To find an optimum induction strategy, effects of various key cultivation variables on the mini-proinsulin production are examined in high cell density fed-batch cultures. No general correlation is found between preinduction specific growth rate and recombinant protein synthesis, which confers a flexibility in choosing the feeding strategy of preinduction media for achieving the high cell density cultures. A culture temperature below 37 degrees C is unfavorable for recombinant gene expression, and the T7-based expression system is almost completely repressed at 30 degrees C. The nutrient glucose and yeast extract concentration in postinduction feed media is optimized by applying a statistical method for medium optimization, i.e. response surface methodology, and an effective amount of inducer molecule (IPTG) is determined to maximize the specific recombinant protein formation. The mini-proinsulin production in E. coli culture is significantly influenced by the volumetric feed rate of postinduction media, which is shown to be closely related to the plasmid copy number in the recombinant cell. Consequently, in a single-stage fed-batch process, the mini-proinsulin concentration is increased up to 7 g/L, approximately 62 wt % of which corresponds to mature human insulin. A two-stage fed-batch fermentation process, with recombinant cell growth occurring at a constant growth rate and constant cell concentration in a growth fermenter and mini-proinsulin production in an induction fermenter, is designed, and its efficacy in increasing volumetric productivity of mini-proinsulin is demonstrated.


Different social motives in the gestural communication of chimpanzees and human children.
Both chimpanzees and human infants use the pointing gesture with human adults, but it is not clear if they are doing so for the same social motives. In two studies, we presented chimpanzees and human 25-month-olds with the opportunity to point for a hidden tool (in the presence of a non-functional distractor). In one condition it was clear that the tool would be used to retrieve a reward for the pointing subject (so the pointing was selfish or 'for-me'), whereas in the other condition it was clear that the tool would be used to retrieve the reward for the experimenter (so the pointing was helpful or 'for-you'). The chimpanzees pointed reliably only when they themselves benefited, whereas the human children pointed reliably no matter who benefited. These results are interpreted as evidence for the especially cooperative nature of human communication.


Clinical applications of dendritic cell vaccination in the treatment of cancer.
Dendritic cell (DC) immunotherapy has shown significant promise in animal studies as a potential treatment for cancer. Its application in the clinic depends on the results of human trials. Here, we review the published clinical trials of cancer immunotherapy using exogenously antigen-exposed DCs. We begin with a short review of general properties and considerations in the design of such vaccines. We then review trials by disease type. Despite great efforts on the part of individual investigative groups, most trials to date have not yielded data from which firm conclusions can be drawn. The reasons for this include nonstandard DC preparation and vaccination protocols, use of different antigen preparations, variable means of immune assessment, and nonrigorous criteria for defining clinical response. While extensive animal studies have been conducted using DCs, optimal parameters in humans remain to be established. Unanswered questions include optimal cell dose, use of mature versus immature DCs for vaccination, optimal antigen preparation, optimal route, and optimal means of assessing immune response. It is critical that these questions be answered, as DC therapy is labor- and resource-intensive. Cooperation is needed on the part of the many investigators in the field to address these issues. If such cooperation is not forthcoming, the critical studies that will be required to make DC therapy a clinically and commercially viable enterprise will not take place, and this therapy, so promising in preclinical studies, will not be able to compete with the many other new approaches to cancer therapy presently in development. Trials published in print through June 2003 are included. We exclude single case reports, except where relevant, and trials with so many variables as to prevent interpretation about DC therapy effects.


The neuronal substrate of risky choice: an insight into the contributions of neuroimaging to the understanding of theories on decision making under risk.
This chapter provides an overview of studies in the field of neuroscience that investigate some of the processes and concepts of risk perception, risky choice, and decision making under risk. First, early studies in the field of neuroscience addressing the diminished decision-making abilities in lesion patients are presented. A classical task in this research field is described along with its neural implications. After this, the underlying model, its hypotheses, and neuronal implications are discussed. Different aspects within risky decision making, such as the influence of memory, inhibition, motivation, and personality, on risky choice and the respective underlying neuronal substrate are described. After this, studies of risky decision making in healthy subjects are reviewed. A selection of studies shows that theories focusing on cognitive aspects only have to be enriched in order to allow for additional aspects within risky decision making (e.g., emotion). Next, the classical economic approaches and the development of theories incorporating further aspects within economical decision making and the underlying neuronal substrate will be presented. Finally, research in the field of neuroeconomics, focusing on the role of social decision making and evaluative judgment within risky decision making, is reviewed.


Inhalational anesthetics up-regulate constitutive and lipopolysaccharide-induced inducible nitric oxide synthase expression and activity.
Nitric oxide (NO) is an important biological messenger involved in the regulation of blood vessel tone, neurotransmission, inflammatory responses, and host defenses. Inhalational anesthetics have been shown to inhibit the function of the NO signaling pathway in a variety of tissues. In addition, acute inhibition of the NO signaling pathway significantly reduced the required alveolar concentration of halothane or isoflurane for anesthesia, which suggests a role for the NO signaling pathway in mechanisms of anesthesia and consciousness. We now report that inhalational anesthetics affect gene expression of nitric oxide synthases (NOS) (EC 220.127.116.11), the enzymes that synthesize NO from L-arginine. Both halothane and isoflurane, at clinically relevant concentrations, significantly up-regulate the mRNA, protein, and activity level of NOS in lipopolysaccharide-treated macrophages (inducible NOS; type II NOS), and bovine pulmonary endothelial cells (endothelial constitutive NOS; type III NOS). This is a novel interaction between inhalational anesthetics and the NO signaling pathway and has wide-ranging implications for both clinical medicine and experimental biology.


Extracellular vesicles shed from gastric cancer mediate protumor macrophage differentiation.
Peritoneal dissemination often develops in gastric cancer. Tumor-associated macrophages (TAMs) are present in the peritoneal cavity of gastric cancer patients with peritoneal dissemination, facilitating tumor progression. However, the mechanism by which macrophages differentiate into tumor-associated macrophages in the peritoneal cavity is not well understood. In this study, the interplay between gastric cancer-derived extracellular vesicles (EVs) and macrophages was investigated. The association between macrophages and EVs in peritoneal ascitic fluid of gastric cancer patients, or from gastric cancer cell lines was examined, and their roles in differentiation of macrophages and potentiation of the malignancy of gastric cancer were further explored. Immunofluorescent assays of the ascitic fluid showed that M2 macrophages were predominant along with the cancer cells in the peritoneal cavity. EVs purified from gastric cancer cells, as well as malignant ascitic fluid, differentiated peripheral blood mononuclear cell-derived macrophages into the M2-like phenotype, which was demonstrated by their morphology and expression of CD163/206. The macrophages differentiated by gastric cancer-derived EVs promoted the migration ability of gastric cancer cells, and the EVs carried STAT3 protein. EVs derived from gastric cancer play a role by affecting macrophage phenotypes, suggesting that this may be a part of the underlying mechanism that forms the intraperitoneal cancer microenvironment.


European Collaboration on Low-dose Aspirin in Polycythemia Vera (ECLAP): a randomized trial.
Thrombotic complications characterize the clinical course of polycythemia vera (PV) and represent the main cause of morbidity and mortality. However, uncertainty still exists as to the benefit/risk ratio of aspirin prophylaxis in this setting. In vivo platelet biosynthesis of thromboxane A2 is enhanced and can be suppressed by low-dose aspirin in PV, thus providing a rationale for assessing the efficacy and safety of a low-dose aspirin regimen in these patients. The Gruppo Italiano Studio Policitemia Vera has recently performed a pilot study on 112 patients randomized to receive aspirin, 40 mg daily, or placebo and followed for 16 +/- 6 months (mean +/- SD). This study showed that low-dose aspirin is well tolerated in PV patients, and that a large-scale efficacy trial is feasible in this setting. In this article we report the protocol of the European Collaboration on Low-dose Aspirin in Polycythemia Vera (ECLAP) study, which is a randomized trial designed to assess the risk/benefit ratio of low-dose aspirin in PV. To estimate the size and the follow-up duration required for the ECLAP trial, a retrospective analysis of the clinical epidemiology of a large PV population has recently been completed by the Gruppo Italiano Studio Policitemia Vera. On this basis, approximately 3500 patients will be enrolled in the ECLAP study with a follow-up of 3 to 4 years. The uncertainty principle will be used as the main eligibility criterion: Polycythemic patients of any age, having no clear indication for or contraindication to aspirin treatment, will be randomized in a double-blind fashion to receive oral aspirin (100 mg daily) or placebo. According to current therapeutic recommendations, the basic treatment of randomized patients should be aimed at maintaining the hematocrit value < or = 45% in subjects aged < or = 50, and hematocrit < 45% as well as platelet count < 400 x 10(9)/L in patients aged > 50. Randomization will be stratified by participating center. The study is funded by the European Union BIOMED 2 program.


Proteolysis Triggers Self-Assembly and Unmasks Innate Immune Function of a Human α-Defensin Peptide.
Human α-defensin 6 (HD6) is a unique peptide of the defensin family that provides innate immunity in the intestine by self-assembling to form high-order oligomers that entrap bacteria and prevent host cell invasion. Here, we report critical steps in the self-assembly pathway of HD6. We demonstrate that HD6 is localized in secretory granules of small intestinal Paneth cells. HD6 is stored in these granules as an 81-residue propeptide (proHD6), and is recovered from ileal lumen as a 32-residue mature peptide. The propeptide neither forms higher-order oligomers, nor agglutinates bacteria, nor prevents Listeria monocytogenes invasion into epithelial cells. The Paneth cell granules also contain the protease trypsin, and trypsin-catalyzed hydrolysis of proHD6 liberates mature HD6, unmasking its latent activities. This work illustrates a remarkable example of how nature utilizes a propeptide strategy to spatially and temporally control peptide self-assembly, and thereby initiates innate immune function in the human intestine.


Oral consumption of Garcinia kola(Bitter kola) lowers intraocular pressure.
Garcinia kola (bitter kola) is locally ingested across the West African subregion. It has ocular hypotensive effects similar to some commonly used glaucoma medications when administered topically. The study assessed the effect of oral ingestion of G. kola on intraocular pressure (IOP). A randomized, single-blind, placebo-controlled, cross-over design was used in this study. Forty-six healthy subjects, aged between 19 and 27 years were recruited and randomized into two groups (A and B). Subjects in group A ingested 100 mg/kg body weight bitter kola in a 200 ml solution on their first visit and group B ingested 200 ml of water. On the second visit, the order of treatment was reversed, IOP was measured at baseline and every 45 min interval for 135 min. The mean difference between the baseline and post-treatment IOP measurements were tested for statistical significance using repeated-measures analysis of variance (95% confidence intervals [CIs]). Mean IOP measurements decreased by 7.9, 18.2 and 20.6% at 45, 90 and 135 min, respectively, after G. kola treatment. The reduction, though variable across subjects, was statistically significant (F [2.13, 95.62] = 90.35, p < 0.0001) across the respective time points. Repetition of an identical protocol without G. kola caused clinically negligible changes in IOP. There was no statistically significant influence of gender or age in G. kola effect on IOP reading. Oral ingestion of G. kola lowered the intraocular pressure of healthy young adults by 21%. Such an effect may be of therapeutic benefit to patients with POAG or ocular hypertension in low-income settings.


Cutaneous melanoma: an audit of management timeliness against New Zealand guidelines.
The New Zealand Ministry of Health's "Faster Cancer Treatment" programme aims for timely care for patients with cancer, including melanoma. Melanoma care guidelines detail investigation and treatment timeliness standards. This audit assesses compliance with these. Patients admitted to Waikato Hospital for melanoma surgery during the year ending February 2016 were retrospectively identified. Time intervals between care events were calculated. Demographic, lesion, surgical and histopathological characteristics were analysed. For patients referred with skin lesions suspicious for melanoma, referral to first treatment (Standard 2.1), referral to diagnostic skin biopsy (Standards 2.2, 2.3), biopsy histology report to first treatment (Standard 2.4), referral to first treatment (Standards 2.2, 2.3, 2.4, 4.4) and biopsy to first treatment (Standards 2.4, 4.4) compliance was 0%, 17.6%, 21.7%, 9.3% and 21.7%, respectively. For patients referred with biopsy-confirmed melanomas, referral to first treatment (Standards 2.2, 2.4) and skin biopsy to first treatment (Standards 2.2, 2.4, 4.4) compliance was 42.2% and 42.9%, respectively. Compliance was low. Attention to logistical constraints in the department reviewed may improve this. Recommendation inconsistencies within and between suspicious-lesion and confirmed-diagnosis referral pathways suggest the investigation and treatment events selected and intervals mandated by the guidelines may usefully be reconsidered.