Training secondary school teachers in instructional language modification techniques to support adolescents with language impairment: a randomized controlled trial.
This study evaluated the efficacy of a collaborative intervention where a speech-language pathologist (SLP) trained mainstream secondary school teachers to make modifications to their oral and written instructional language. The trained teachers' uptake of techniques in their whole-class teaching practices and the impact this had on the language abilities of students with language impairment (LI) were evaluated. Two secondary schools were randomly assigned to either a trained or a control condition. A cohort of 13 teachers (7 trained and 6 control) and 43 Year 8 students with LI (21 trained and 22 control) were tested at pre, post, and follow-up times-teachers by structured interview and students by standardized spoken and written language assessments. Significantly increased use of the language modification techniques by the trained teachers was observed when compared to the control group of untrained teachers, with this increased use maintained over time. Results from the trained group of students showed a significant improvement in written expression and listening comprehension relative to the control group of students. This randomized controlled trial is one of the first investigations to evaluate a collaborative intervention that links changes in mainstream secondary teachers' instructional language practices with improvements in the language abilities of adolescents with LI.


Implications of California's new mental health law.
The author describes the impact that implementation of California's new mental health law and of its amendments has had on the development of community mental health programs. The new legislation has acted as a catalyst to the growth of local mental health programs by providing restrictions on involuntary treatment as well as fiscal incentives for local programs. The effect of the legislation on patients has varied according to patient needs and the resources available in the community. Although the problems and controversies have arisen regarding the placement of chronic patients, impact on law enforcement, and quality of care, the service delivery system developed in response to the new legislation has resulted in a greater emphasis on crisis intervention, greater visibility of direct services, and increased community awareness and concern regarding mental health. The author concludes that mental health laws can become the catalyst for major changes in the mental health services delivery system.


Expression of p21WAF1/CIP1, p53, bcl-2 and Ki-67 in venous leg ulcers with and without squamous cell carcinoma.
In order to gain understanding of the cellular mechanisms of malignant transformation in chronic venous leg ulcers, we analysed by immunohistochemistry the presence of p21, p53, bcl-2 and Ki-67 in ulcers with and without squamous cell carcinoma. The material consisted of 41 archived histopathological samples from 33 patients with chronic venous leg ulcers and 28 samples from 21 patients suffering from squamous cell carcinoma in chronic venous leg ulcers. All samples derived from the chronic ulcers were negative for p53, p21 and bcl-2. Cells positive for Ki-67 were seen in certain ulcer areas. In the cancers, 14 samples showed immunopositivity for p53 and 22 samples were positive for p21, with expression mainly in the suprabasal layer. Expression of p21 seemed independent of p53. None of the samples was bcl-2-positive. Ki-67 was positive with basal expression in 16/28 of the tumour samples, 15 of which also expressed p21. There was no perilesional p53 or p21 activity in the cancer samples. The results regarding perilesional expression are different from those reported in UV-induced squamous cell carcinomas and probably reflect a different carcinogen.


Leaving no hole unclosed: left atrial appendage occlusion in patients having closure of patent foramen ovale or atrial septal defect.
To report procedural outcome of sequential occlusion of the left atrial appendage (LAA) and an interatrial septal communication and discuss possible indications. There are some patients who may have indications for both closure of patent foramen ovale (PFO) or atrial septal defect (ASD), as well as closure of the LAA. The optimal procedural strategy is not known. A retrospective review of LAA and PFO/ASD cases at our center was performed. Demographic, echocardiographic, and procedural data were recorded. Closure of LAA and then PFO/ASD (Group I: 11 patients), closure of both in the same setting (Group II: 3 patients), and closure of the PFO/ASD and then the LAA (Group III: 3 patients) was performed in a total of 17 patients. Average age was 63.5 ± 9.8 years. Most patients were hypertensive with prior cerebrovascular event in 52.9% of patients. Procedural success was 100%. Procedural adverse events were 2 episodes of tamponade (in Group I after first LAA procedure) treated with pericardiocentesis and 1 access-site hematoma (Group II) treated conservatively. Sequential (staged or during the same procedure) closure of the LAA and interatrial communications can be performed safely in a carefully selected patient population. This is also the first known report of LAA occlusion in patients with prior septal closure devices.


Towards the development of a reflective radiographer: challenges and constraints.
Currently there is overwhelming support from the health professions for universities to devise curricular approaches that lead to the development of undergraduate reflective skills, and over time, reflective practice. However, in the case of radiography, irrespective of the degree to which radiographers might engage in reflective practice they constantly struggle to shed the perception they are little more than technical operatives. The four-year Bachelor of Radiography and Medical Imaging was introduced by Monash University in 1998 with an overt commitment to the development of a reflective radiographer. Findings from student and supervisor surveys generally support the program and its aims. However, as the findings and student feedback will attest, many challenges and constraints continue to face educators who seek to situate their curriculum within the reflective practice paradigm.


[Revascularization or conservative strategy in patients with stable coronary heart disease: a contemporary view].
Choice between revascularization and conservative strategy in patients with stable coronary heart disease (CHD) is an important clinical problem, especially when the need for revascularization is not urged by severe ischemic symptoms. Revascularization aimed to improve prognosis is most justified in patients having high risk, based both on results of coronary angiography and functional tests. In the opinion of many experts, the high-risk features at stress-test is a key to identify patients in whom revascularization is most required among persons with high-risk coronary anatomy, as well as to select candidates for invasive treatment among patients with more favorable variants of coronary lesions. The advantage of revascularization over conservative treatment was demonstrated primarily in relation to coronary bypass surgery; however, the prospect of replacing surgery by an interventional treatment for prognostic reasons is extremely attractive. Although comparison of prognostic efficiency of percutaneous coronary intervention (PCI) and coronary bypass surgery (CABG) for some categories of patients with stable ischemic heart disease has yielded encouraging results, improved survival when using the PCI in direct comparison with optimal medical therapy was not convincingly confirmed to date. Implementation of PCI to improve prognosis in stable ischemic heart disease is still limited by a rather narrow range of indications and the relief of symptoms of ischemia remains its main purpose.


An Artificial Enzyme Made by Covalent Grafting of an Fe(II) Complex into β-Lactoglobulin: Molecular Chemistry, Oxidation Catalysis, and Reaction-Intermediate Monitoring in a Protein.
An artificial metalloenzyme based on the covalent grafting of a nonheme Fe(II) polyazadentate complex into bovine β-lactoglobulin has been prepared and characterized by using various spectroscopic techniques. Attachment of the Fe(II) catalyst to the protein scaffold is shown to occur specifically at Cys121. In addition, spectrophotometric titration with cyanide ions based on the spin-state conversion of the initial high spin (S=2) Fe(II) complex into a low spin (S=0) one allows qualitative and quantitative characterization of the metal center's first coordination sphere. This biohybrid catalyst activates hydrogen peroxide to oxidize thioanisole into phenylmethylsulfoxide as the sole product with an enantiomeric excess of up to 20 %. Investigation of the reaction between the biohybrid system and H2 O2 reveals the generation of a high spin (S=5/2) Fe(III) (η(2) -O2 ) intermediate, which is proposed to be responsible for the catalytic sulfoxidation of the substrate.


Hyperoxia may be related to delayed cerebral ischemia and poor outcome after subarachnoid haemorrhage.
To determine the association between exposure to hyperoxia and the risk of delayed cerebral ischaemia (DCI) after subarachnoid haemorrhage (SAH). We analysed data from a single centre, prospective, observational cohort database. Patient inclusion criteria were age ≥18 years, aneurysmal SAH, endotracheal intubation with mechanical ventilation, and arterial partial pressure of oxygen (PaO2) measurements. Hyperoxia was defined as the highest quartile of an area under the curve of PaO2, until the development of DCI (PaO2≥173 mm Hg). Poor outcome was defined as modified Rankin Scale 4-6 at 3 months after SAH. Of 252 patients, there were no differences in baseline characteristics between the hyperoxia and control group. Ninety-seven (38.5%) patients developed DCI. The hyperoxia group had a higher incidence of DCI (p<0.001) and poor outcome (p=0.087). After adjusting for modified Fisher scale, rebleeding, global cerebral oedema, intracranial pressure crisis, pneumonia and sepsis, hyperoxia was independently associated with DCI (OR, 3.16; 95% CI 1.69 to 5.92; p<0.001). After adjusting for age, Hunt-Hess grade, aneurysm size, Acute Physiology and Chronic Health Evaluation II score, rebleeding, pneumonia and sepsis, hyperoxia was independently associated with poor outcome (OR, 2.30; 95% CI 1.03 to 5.12; p=0.042). In SAH patients, exposure to hyperoxia was associated with DCI. Our findings suggest that exposure to excess oxygen after SAH may represent a modifiable factor for morbidity and mortality in this population.


Factors associated with pediatric use of complementary and alternative medicine.
Limited data are available on the use of complementary and alternative medicine (CAM) and factors associated with use among the pediatric population in the United States. Using the 2007 National Health Interview Survey data among individuals <18 years of age (n = 9417), we compared CAM users (excluding those using vitamins and minerals) and non-CAM users. Using bivariable and multivariable logistic regression models, we examined independent associations of CAM use with sociodemographic factors, prescription medication use, delays in health care caused by access difficulties, and common medical conditions/symptoms. In an adjusted multivariable logistic model, CAM users were more likely than non-CAM users to be adolescents rather than infants or toddlers (adjusted odds ratio [aOR]: 1.61 [95% confidence interval (CI): 1.11-2.34]); live in the West (aOR: 2.05 [95% CI: 1.62-2.59]), Northeast (aOR: 1.36 [95% CI: 1.02-1.80]), or Midwest (aOR: 1.35 [95% CI: 1.04-1.74]) compared with those in the South; more likely to have a parent with a college education (aOR: 4.33 [95% CI: 2.92-6.42]); and more likely to use prescription medication (aOR: 1.51 [95% CI: 1.19-1.92]). Pediatric CAM users were more likely to have anxiety or stress (aOR: 2.54 [95% CI: 1.89-3.42]), dermatologic conditions (aOR: 1.35 [95% CI: 1.03-1.78]), musculoskeletal conditions (aOR: 1.94 [95% CI: 1.31-2.87]), and sinusitis (aOR: 1.54 [95% CI: 1.11-2.14]). Use of CAM by a parent was strongly associated with the child's use of CAM (aOR: 3.83 [95% CI: 3.04-4.84]). In 2007, pediatric CAM users were more likely to take prescription medications, have a parent who used CAM, and have chronic conditions such as anxiety or stress, musculoskeletal conditions, dermatologic conditions, or sinusitis. Research is required to guide pediatricians in making recommendations on CAM modalities for children including potential risks and/or benefits and interactions with conventional therapies.


Complications From Tunneled Hemodialysis Catheters: A Canadian Observational Cohort Study.
Clinical practice guidelines discourage the use of central venous catheters (CVCs) for vascular access in dialysis. However, some patients have inadequate vessels for arteriovenous fistula creation or choose to use a dialysis catheter. The risks associated with CVC use and their relationship to patient age are poorly characterized. Observational retrospective cohort study. Cohort of 1,041 patients older than 18 years from 5 Canadian dialysis programs who initiated outpatient maintenance hemodialysis therapy with a tunneled CVC between 2004 and 2012 and who had no creation of an arteriovenous fistula or arteriovenous graft. Age, sex, body size, initiating dialysis therapy in the hospital, and comorbid conditions. CVC-related procedures, hospitalization, and death. Complications were reported as a cumulative risk at 1 and 2 years. Cox proportional hazards regression for recurrent events was used to evaluate risk factors for study outcomes. At 1 year, risks for CVC-related bacteremia, malfunction, and central stenosis were 9%, 15%, and 2%, respectively. Risks for any CVC-related complication at 1 and 2 years were 30% and 38%, respectively. Death related to CVC complications occurred in 6 of 1,041 (0.5%) patients. Compared with patients younger than 60 years, patients aged 70 to 79 and those 80 years or older experienced lower rates of CVC complications: HRs of 0.67 (95% CI, 0.52-0.85; P = 0.001) and 0.69 (95% CI, 0.52-0.92; P = 0.01), respectively. This Canadian dialysis population may not be representative of populations in other countries. CVC use was not compared with other types of hemodialysis vascular access. Approximately one-third of hemodialysis patients who used tunneled CVCs during 1 to 2 years experienced complications. Bacteremia occurred in ∼9% of patients at 1 year and were the most common cause of CVC-related hospitalizations. CVC-related death was infrequent. This information could be used to communicate the risk for CVC complications to patients treated with this type of hemodialysis vascular access.


Chemical imaging in a surface forces apparatus: confocal raman spectroscopy of confined poly(dimethylsiloxane).
Confocal Raman spectroscopy has been implemented within the molecularly thin films of a surface forces apparatus. Applying this technique to an initial system, we investigate the confinement and shear-induced changes in the Raman spectra of poly(dimethylsiloxane) (PDMS) liquids confined between atomically smooth mica surfaces at thicknesses less than the unperturbed radius of gyration of the polymer. We focus on stretch vibrations of the PDMS methyl group, whose net orientation is perpendicular to the chain backbone. When PDMS was confined to a thickness of approximately the unperturbed radius of gyration (3.5 nm) but no shear, the Raman intensity of the methyl group was anisotropic in the x-y plane, signifying that chains oriented preferentially parallel to the confining surfaces. Relative to the bulk fluid, the relative intensity of the asymmetric to symmetric carbon-hydrogen stretch (2965 and 2907 cm-1, respectively) was enhanced, indicating that asymmetric vibration was enhanced by confinement. Measurements using polarized radiation showed coherent planar anisotropy in the x-y plane whose direction varied stochastically from experiment to experiment. It seems that although coherent in-plane alignment was favored, no preferential alignment direction was favored in the absence of shear. Application of shear caused the time-averaged polymer conformations to become more nearly isotropic in the plane of shear. These measurements are considered to represent the first chemical imaging of chemical species within the contact area of a surface forces apparatus.


Arsenic trioxide-induced apoptosis through oxidative stress in cells of colon cancer cell lines.
Exposure of three colon cancer cell lines, SW480, DLD-1, and COLO201, to arsenic trioxide in the medium induced a marked concentration-dependent suppression of cell growth. The intracellular contents of reduced glutathione (GSH) in these cell lines tended to be inversely correlated with the sensitivity of the cells to arsenic trioxide. Among the cell lines, SW480 cells underwent apoptosis at the low arsenic trioxide concentration of 2 microM, which was prevented by pretreatment of the cells with N-acetylcysteine and was enhanced by buthionine sulfoximine. The production of reactive oxygen intermediates which were examined by 2',7'-dichlorodihydrofluorescein diacetate (H2DCF-DA), increased with time after treatment with arsenic trioxide. The apoptosis was executed by the activation of caspase 3, which was shown by Western blot, enzymatic activity, and apoptosis inhibition assay. The mitochondrial membrane potential of adherent apoptotic SW480 cells and the cells from intermediate layer separated by density gradient centrifugation, both of which showed the active form of caspase 3 by Western blot analysis, was not lost. The overexpression of Bcl-2 protein in SW480 cells could not prevent the apoptosis induced by the treatment with arsenic trioxide. All these findings indicate that arsenic trioxide-induced apoptosis in SW480 cells is executed by the activation of caspase 3 without mediating by mitochondria under the overproduction of reactive oxygen species.


The biological hallmarks of ileal carcinoids.
Endocrine tumours derived from the small intestine, ileal carcinoids, produce and secrete the hormones tachykinins and serotonin, which induces the specific symptoms related to the tumour. Because of their low proliferation rate, they are often discovered at late stages when metastases have occurred. The biology that characterizes these tumours differs in many ways from what is generally recognized for other malignancies. In this overview, the current knowledge on the development and progression of ileal carcinoids is described.


Minoxidil is a potential neuroprotective drug for paclitaxel-induced peripheral neuropathy.
Chemotherapy-induced peripheral neuropathy (CIPN) is a common side effect of cancer treatment. No medication has been shown to be effective in the treatment of CIPN. This study aims to integrate the image-based high-content screening, mouse behavior models and mechanistic cell-based assays to discover potential neuroprotective drugs. Among screened compounds, minoxidil showed the most potent neuroprotective effect against paclitaxel, with regard to neurite outgrowth of dorsal root ganglia (DRG). Minoxidil protected mice from thermal insensitivity and alleviated mechanical allodynia in paclitaxel-treated mice. The ultrastructure and quantified G-ratio of myelin integrity of sciatic nerve tissues supported the observations in mouse behavioral tests. The mechanistic study on DRG neurons suggested that minoxidil suppressed neuroinflammation and remodeled the dysregulation of intracellular calcium homeostasis provoked by paclitaxel. Importantly, minoxidil showed a synergistic anti-tumor effect with paclitaxel both in tumor xenograft models of cervical and breast cancer. Interestingly, the quantitative assays on hair length and hair growth both exhibited that minoxidil significantly improved the hair quality after chemotherapy. Since minoxidil is a drug approved by the Food and Drug Administration (FDA), the safety and biocompatibility are well documented. The immediate next step is to launch an early-stage clinical trial intending to prevent CIPN by minoxidil.


Activation of the pseudokinase MLKL unleashes the four-helix bundle domain to induce membrane localization and necroptotic cell death.
Necroptosis is considered to be complementary to the classical caspase-dependent programmed cell death pathway, apoptosis. The pseudokinase Mixed Lineage Kinase Domain-Like (MLKL) is an essential effector protein in the necroptotic cell death pathway downstream of the protein kinase Receptor Interacting Protein Kinase-3 (RIPK3). How MLKL causes cell death is unclear, however RIPK3-mediated phosphorylation of the activation loop in MLKL trips a molecular switch to induce necroptotic cell death. Here, we show that the MLKL pseudokinase domain acts as a latch to restrain the N-terminal four-helix bundle (4HB) domain and that unleashing this domain results in formation of a high-molecular-weight, membrane-localized complex and cell death. Using alanine-scanning mutagenesis, we identified two clusters of residues on opposing faces of the 4HB domain that were required for the 4HB domain to kill cells. The integrity of one cluster was essential for membrane localization, whereas MLKL mutations in the other cluster did not prevent membrane translocation but prevented killing; this demonstrates that membrane localization is necessary, but insufficient, to induce cell death. Finally, we identified a small molecule that binds the nucleotide binding site within the MLKL pseudokinase domain and retards MLKL translocation to membranes, thereby preventing necroptosis. This inhibitor provides a novel tool to investigate necroptosis and demonstrates the feasibility of using small molecules to target the nucleotide binding site of pseudokinases to modulate signal transduction.


Characterization and drug resistance patterns of Ewing's sarcoma family tumor cell lines.
Despite intensive treatment with chemotherapy, radiotherapy and surgery, over 70% of patients with metastatic Ewing's Sarcoma Family of Tumors (EFT) will die of their disease. We hypothesize that properly characterized laboratory models reflecting the drug resistance of clinical tumors will facilitate the application of new therapeutic agents to EFT. To determine resistance patterns, we studied newly established EFT cell lines derived from different points in therapy: two established at diagnosis (CHLA-9, CHLA-32), two after chemotherapy and progressive disease (CHLA-10, CHLA-25), and two at relapse after myeloablative therapy and autologous bone marrow transplantation (post-ABMT) (CHLA-258, COG-E-352). The new lines were compared to widely studied EFT lines TC-71, TC-32, SK-N-MC, and A-673. These lines were extensively characterized with regard to identity (short tandem repeat (STR) analysis), p53, p16/14 status, and EWS/ETS breakpoint and target gene expression profile. The DIMSCAN cytotoxicity assay was used to assess in vitro drug sensitivity to standard chemotherapy agents. No association was found between drug resistance and the expression of EWS/ETS regulated genes in the EFT cell lines. No consistent association was observed between drug sensitivity and p53 functionality or between drug sensitivity and p16/14 functionality across the cell lines. Exposure to chemotherapy prior to cell line initiation correlated with drug resistance of EFT cell lines in 5/8 tested agents at clinically achievable concentrations (CAC) or the lower tested concentration (LTC): (cyclophosphamide (as 4-HC) and doxorubicin at CAC, etoposide, irinotecan (as SN-38) and melphalan at LTC; P<0.1 for one agent, and P<0.05 for four agents. This panel of well-characterized drug-sensitive and drug-resistant cell lines will facilitate in vitro preclinical testing of new agents for EFT.


Visualizing viral protein structures in cells using genetic probes for correlated light and electron microscopy.
Structural studies of viral proteins most often use high-resolution techniques such as X-ray crystallography, nuclear magnetic resonance, single particle negative stain, or cryo-electron microscopy (EM) to reveal atomic interactions of soluble, homogeneous viral proteins or viral protein complexes. Once viral proteins or complexes are separated from their host's cellular environment, their natural in situ structure and details of how they interact with other cellular components may be lost. EM has been an invaluable tool in virology since its introduction in the late 1940's and subsequent application to cells in the 1950's. EM studies have expanded our knowledge of viral entry, viral replication, alteration of cellular components, and viral lysis. Most of these early studies were focused on conspicuous morphological cellular changes, because classic EM metal stains were designed to highlight classes of cellular structures rather than specific molecular structures. Much later, to identify viral proteins inducing specific structural configurations at the cellular level, immunostaining with a primary antibody followed by colloidal gold secondary antibody was employed to mark the location of specific viral proteins. This technique can suffer from artifacts in cellular ultrastructure due to compromises required to provide access to the immuno-reagents. Immunolocalization methods also require the generation of highly specific antibodies, which may not be available for every viral protein. Here we discuss new methods to visualize viral proteins and structures at high resolutions in situ using correlated light and electron microscopy (CLEM). We discuss the use of genetically encoded protein fusions that oxidize diaminobenzidine (DAB) into an osmiophilic polymer that can be visualized by EM. Detailed protocols for applying the genetically encoded photo-oxidizing protein MiniSOG to a viral protein, photo-oxidation of the fusion protein to yield DAB polymer staining, and preparation of photo-oxidized samples for TEM and serial block-face scanning EM (SBEM) for large-scale volume EM data acquisition are also presented. As an example, we discuss the recent multi-scale analysis of Adenoviral protein E4-ORF3 that reveals a new type of multi-functional polymer that disrupts multiple cellular proteins. This new capability to visualize unambiguously specific viral protein structures at high resolutions in the native cellular environment is revealing new insights into how they usurp host proteins and functions to drive pathological viral replication.


Identification and expression of maebl, an erythrocyte-binding gene, in Plasmodium gallinaceum.
Avian malaria is of significant ecological importance and serves as a model system to study broad patterns of host switching and host specificity. The erythrocyte invasion mechanism of the malaria parasite Plasmodium is mediated, in large part, by proteins of the erythrocyte-binding-like (ebl) family of genes. However, little is known about how these genes are conserved across different species of Plasmodium, especially those that infect birds. Using bioinformatical methods in conjunction with polymerase chain reaction (PCR) and genetic sequencing, we identified and annotated one member of the ebl family, merozoite apical erythrocyte-binding ligand (maebl), from the chicken parasite Plasmodium gallinaceum. We then detected the expression of maebl in P. gallinaceum by PCR analysis of cDNA isolated from the blood of infected chickens. We found that maebl is a conserved orthologous gene in avian, mammalian, and rodent Plasmodium species. The duplicate extracellular binding domains of MAEBL, responsible for erythrocyte binding, are the most conserved regions. Our combined data corroborate the conservation of maebl throughout the Plasmodium genus and may help elucidate the mechanisms of erythrocyte invasion in P. gallinaceum and the host specificity of Plasmodium parasites.


Immunostaining for detecting lymph node metastases in non-small cell lung cancer.
To detect lymph node metastases by immunohistochemistry, where previously undetected by routine histopathology. Immunostaining was carried out for high- and low molecular weight cytokeratins, and Ber-EP4 in 19 consecutive lung cancer patients who had undergone systematic mediastinal lymph node dissection. Eleven (58%) epidermoid carcinomas, 6 (32%) adenocarcinomas, and 2 (10%) bronchiolo-alveolar carcinomas were detected. These included 4 (21%) stage IA carcinomas, 6 (32%) stage IB, 6 (32%) stage IIB, 1 (5%) stage IIIB and 2 (10%) stage IV. Immunostaining did not reveal any undetected metastases. Two patients (squamous cell carcinoma T1N0; adenocarcinoma T1N0) had metastases (skeletal; ipsilateral lung) at time of surgery, and one patient (squamous cell carcinoma T2N0) had a regional and systemic relapse 10 months later. Serial sectioning with immunostaining of the lymph nodes from these three patients was also negative. We conclude that, even with the use of immunostaining, negative lymph nodes will not assure a good prognosis, and different determinants probably exist for lymphatic and hematogenic metastases in non-small cell lung cancer.


miR-34 is maternally inherited in Drosophila melanogaster and Danio rerio.
MicroRNAs (miRNAs) are small, endogenous, regulatory RNA molecules that can bind to partially complementary regions on target messenger RNAs and impede their expression or translation. We rationalized that miRNAs, being localized to the cytoplasm, will be maternally inherited during fertilization and may play a role in early development. Although Dicer is known to be essential for the transition from single-celled zygote to two-cell embryo, a direct role for miRNAs has not yet been demonstrated. We identified miRNAs with targets in zygotically expressed transcripts in Drosophila using a combination of transcriptome analysis and miRNA target prediction. We experimentally established that Drosophila miRNA dme-miR-34, the fly homologue of the cancer-related mammalian miRNA miR-34, involved in somatic-cell reprogramming and having critical role in early neuronal differentiation, is present in Drosophila embryos before initiation of zygotic transcription. We also show that the Drosophila miR-34 is dependent on maternal Dicer-1 for its expression in oocytes. Further, we show that miR-34 is also abundant in unfertilized oocytes of zebrafish. Its temporal expression profile during early development showed abundant expression in unfertilized oocytes that gradually decreased by 5 days post-fertilization (dpf). We find that knocking down the maternal, but not the zygotic, miR-34 led to developmental defects in the neuronal system during early embryonic development in zebrafish. Here, we report for the first time, the maternal inheritance of an miRNA involved in development of the neuronal system in a vertebrate model system.


Liquid biopsies: genotyping circulating tumor DNA.
Genotyping tumor tissue in search of somatic genetic alterations for actionable information has become routine practice in clinical oncology. Although these sequence alterations are highly informative, sampling tumor tissue has significant inherent limitations; tumor tissue is a single snapshot in time, is subject to selection bias resulting from tumor heterogeneity, and can be difficult to obtain. Cell-free fragments of DNA are shed into the bloodstream by cells undergoing apoptosis or necrosis, and the load of circulating cell-free DNA (cfDNA) correlates with tumor staging and prognosis. Moreover, recent advances in the sensitivity and accuracy of DNA analysis have allowed for genotyping of cfDNA for somatic genomic alterations found in tumors. The ability to detect and quantify tumor mutations has proven effective in tracking tumor dynamics in real time as well as serving as a liquid biopsy that can be used for a variety of clinical and investigational applications not previously possible.


Argentometric titration for the determination of liquid chromatographic injection reproducibility.
The reproducibility of autosamplers for high-performance liquid chromatography can be checked by the injection of a sodium chloride solution without an installed column. The dispensed chloride plug is analyzed quantitatively by argentometric titration. If the titration can be performed with high enough precision, the autosampler performance is investigated without additional effects. Experiments with a 20-microL loop were performed. We found the following repeatabilities (relative standard deviations): titration, 0.07%; 10-microL "pull loop" injection (partial filling of the loop), 0.6%; and 20-microL "full loop" injection, 0.1%. In contrast to these data, the relative standard deviations obtained from chromatographic peaks include the steps of injection, separation, detection, and integration without any knowledge of the individual contributions. Such values are higher; we found for chromatographic peak areas of parabens 2% under pull loop conditions and 0.17% by full loop. The titration approach allows the determination of the contribution of the injection to the uncertainty budget of a liquid chromatographic analysis.


Human dioxin-inducible cytochrome P1-450: complementary DNA and amino acid sequence.
Induction of cytochrome P1-450 has been linked to susceptibility to certain chemically induced cancers in mouse and man. Treatment of the human cell line MCF-7 with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) results in high levels of aryl hydrocarbon (benzo[a]pyrene) hydroxylase (P1-450) activity. This cell line was used to isolate a human P1-450 full-length complementary DNA (cDNA) clone. The cDNA is 2566 nucleotides in length, encodes a polyadenylated messenger RNA (2.8 kilobases in length), and has a continuous reading frame producing a protein with 512 residues (molecular weight, 58,151). The human P1-450 cDNA and protein are 63 percent and 80 percent similar to mouse P1-450 cDNA and protein, respectively. Whereas the mouse TCDD-inducible P-450 gene subfamily has two members (P1-450 and P3-450), the human TCDD-inducible gene subfamily appears to have only one gene (P1-450).


Highly flexible anisotropic magnetoresistance sensor for wearable electronics.
The emerging market of wearable devices for tracking and positioning requires the development of highly flexible magnetic sensors. Due to the stable magnetoresistance ratio and simple fabrication process, sensors based on the anisotropic magnetoresistance (AMR) effect have been proposed as promising candidates. However, the stability of AMR sensors under different bending situations has rarely been investigated. In this work, we proposed a flexible AMR magnetic sensor deposited on an ultra-thin Kapton substrate, which exhibits excellent anti-fatigue behavior at different bending curvatures ranging from 1/3 to 1/10 mm-1. Experimental results show that the sensitivity of our proposed flexible AMR sensor remains as high as 0.25 Oe-1 after being bent 500 times, showing promising potential for application in wearable electronic devices.


N-terminal strands of filamin Ig domains act as a conformational switch under biological forces.
Conformational changes of filamin A under stress have been postulated to play crucial roles in signaling pathways of cell responses. Direct observation of conformational changes under stress is beyond the resolution of current experimental techniques. On the other hand, computational studies are mainly limited to either traditional molecular dynamics simulations of short durations and high forces or simulations of simplified models. Here we perform all-atom discrete molecular dynamics (DMD) simulations to study thermally and force-induced unfolding of filamin A. The high conformational sampling efficiency of DMD allows us to observe force-induced unfolding of filamin A Ig domains under physiological forces. The computationally identified critical unfolding forces agree well with experimental measurements. Despite a large heterogeneity in the population of force-induced intermediate states, we find a common initial unfolding intermediate in all the Ig domains of filamin, where the N-terminal strand unfolds. We also study the thermal unfolding of several filamin Ig-like domains. We find that thermally induced unfolding features an early-stage intermediate state similar to the one observed in force-induced unfolding and characterized by the N-terminal strand being unfurled. We propose that the N-terminal strand may act as a conformational switch that unfolds under physiological forces leading to exposure of cryptic binding sites, removal of native binding sites, and modulating the quaternary structure of domains.


Mechanisms linking traffic-related air pollution and atherosclerosis.
Recent discoveries in the field of air pollution toxicology highlight the potential impact of specific sources of air pollution, especially related to roadway emissions, on acute and chronic cardiovascular disease. This review covers potential mechanisms, both in terms of biological pathways and chemical drivers, to explain these observations. Air pollution is associated with chronic progression of cardiovascular disease. Roadway exposures appear to have a strong correlation to these adverse outcomes. Controlled toxicological studies highlight potential interactions between vehicle-source emissions and adverse vascular outcomes. Mechanistically, a role for both innate and adaptive immune responses is emerging, with important recent findings demonstrating that immunomodulatory pattern-recognition receptors such as Toll-like receptor-4 and lectin-like oxidized LDL receptor-1 may play a role in communicating airway exposures to cardiovascular outcomes. An improved understanding of the sources and mechanisms underlying adverse cardiovascular health outcomes of air pollution would enhance our ability to manage vulnerable populations and establish precise, effective regulatory policies.


Iterons Homologous to Helper Geminiviruses Are Essential for Efficient Replication of Betasatellites.
Betasatellites associated with geminiviruses can be replicated promiscuously by distinct geminiviruses but exhibit a preference for cognate helper viruses. However, the cis elements responsible for betasatellite origin recognition have not been characterized. In this study, we identified an iteron-like repeated sequence motif, 5'-GAGGACC-3', in a tobacco curly shoot betasatellite (TbCSB) associated with tobacco curly shoot virus (TbCSV). Competitive DNA binding assays revealed that two core repeats (5'-GGACC-3') are required for specific binding to TbCSV Rep; TbCSB iteron mutants accumulated to greatly reduced levels and lost the cognate helper-mediated replication preference. Interestingly, TbCSV also contains identical repeated sequences that are essential for specific Rep binding and in vivo replication. In order to gain insight into the mechanism by which TbCSB has acquired the cognate iterons, we performed a SELEX (systematic evolution of ligands by exponential enrichment) assay to identify the high-affinity Rep binding ligands from a large pool of randomized sequences. Analysis of SELEX winners showed that all of the sequences contained at least one core iteron-like motif, suggesting that TbCSB has evolved to contain cognate iterons for high-affinity Rep binding. Further analyses of various betasatellite sequences revealed a region upstream of the satellite conserved region replete with iterative sequence motifs, including species-specific repeats and a general repeat (5'-GGTAAAT-3'). Remarkably, the species-specific repeats in many betasatellites are homologous to those in their respective cognate helper begomoviruses, whereas the general repeat is widespread in most of the betasatellite molecules analyzed. These data, taken together, suggest that many betasatellites have evolved to acquire homologous iteron-like sequences for efficient replication mediated by cognate helper viruses.IMPORTANCE The geminivirus-encoded replication initiator protein (Rep) binds to repeated sequence elements (also known as iterons) in the origin of replication that serve as essential cis elements for specific viral replication. Betasatellites associated with begomoviruses can be replicated by cognate or noncognate helper viruses, but the cis elements responsible for betasatellite origin recognition have not been characterized. Using a betasatellite (TbCSB) associated with tobacco curly shoot virus (TbCSV) as a model, we identify two tandem repeats (iterons) in the Rep-binding motif (RBM) that are required for specific Rep binding and efficient replication, and we show that identical iteron sequences present in TbCSV are also necessary for Rep binding and the replication of helper viruses. Extensive analysis of begomovirus/betasatellite sequences shows that many betasatellites contain iteron-like elements homologous to those of their respective cognate helper begomoviruses. Our data suggest that many betasatellites have evolved to acquire homologous iteron-like sequences for efficient replication mediated by cognate helper viruses.


Phrenic nerve transfer in the repair of brachial plexus injuries: an animal model.
Ten young mongrel dogs underwent unilateral denervation of the brachial plexus. In six dogs, a 2-cm segment of phrenic nerve autograft was sutured to either the resected musculocutaneous nerve or the radial nerve. A hemoclip was applied to either musculocutaneous or radial nerve in the control groups. Five months postoperatively, the grafted musculocutaneous nerve demonstrated less fibrous tissue and less muscle atrophy of the biceps when compared to the control group with clipped nerve. In the group with the grafted radial nerve, the electromyographic findings of multiphasic action potential and muscle contraction from electric stimulation suggested reinnervation of the radial nerve. phrenic nerve transfer may be used to repair specific damages to nerve trunk with histological, electromyographic and clinical recovery.


Diagnostic imaging in COPD.
Chronic obstructive pulmonary disease (COPD) is a pathological pulmonary condition characterized by expiratory airflow obstruction due to emphysematous destruction of the lung parenchyma and small airways remodeling. Although spirometry is a very useful diagnostic tool for screening large groups of smokers, it cannot readily differentiate the etiologies of COPD and thus has limited utility in characterizing subjects for clinical and investigational purposes. There has been a longstanding interest in thoracic imaging and its role in the in vivo characterization of smoking-related lung disease. Research in this area has spanned readily available modalities such as chest -ray and computed tomography to more advanced imaging techniques such as optical coherence tomography (OCT) and magnetic resonance imaging (MRI). Although the chest x-ray is almost universally available, it lacks sensitivity in detecting both airway disease and mild emphysema and is not generally amenable to objective analysis. Computed tomography has become the standard modality to objectively visualize lung disease. It can provide useful measures of the presence and extent of emphysema, airway disease, and, more recently, pulmonary vascular disease for clinical correlation. It does, however, face limitations in standardization across brands and generations of scanners, and the ionizing radiation associated with image acquisition is of concern to both patients and health care providers. Newer techniques such as OCT and MRI offer exciting in vivo insights into lung structure and function that were previously available only in necropsy specimens and physiology laboratories. Given the more limited availability of these techniques, they will be viewed here as adjuncts to computed tomographic imaging.


Can Patients with Pancreatic Cancer and Liver Metastases Obtain Survival Benefit from Surgery? A Population-Based Study.
Background: Surgery for pancreatic cancer with liver metastases (PCL) is not recommended in the international guidelines, and investigation of its clinical significance in patients with PCL is very limited. This study explored whether surgery, especially synchronous resection of the primary tumor and liver metastases (SPL), could improve survival in PCL. Methods: Data of 14,248 patients with PCL from Surveillance, Epidemiology, and End Results database was analyzed. Patients were divided into following groups: SPL, synchronous primary site, and other resection (SPO), single resection of the primary site (SPS), and no resection (NR). Results: In this study, only 93 (0.7%) underwent SPL, 88 (0.6%) for SPO, and 232 (1.6%) for SPS. Multivariate Cox analysis showed surgical procedures of both the primary site and other sites were independent protective prognostic factors for pancreatic cancer cause-specific survival (PCSS) (all P < 0.001). Patients in the SPL group showed the most survival benefit, with a significant and gradually increased difference as compared with the SPO, SPS, and NR groups (median survival: 54, 34, 15, and 3 months, respectively, all P < 0.001). Compared with the NR group, mortalities were significant and gradually declining in the SPS, SPO, and SPL groups, with hazard ratio 0.329 (95% confidence interval [CI], 0.281 to 0.386), 0.220 (95% CI, 0.164 to 0.294), and 0.162 (95% CI, 0.118 to 0.222), respectively (all P < 0.001). Conclusions: Surgical procedures for both primary site and other sites improved survival. SPL, particularly, showed a considerable survival benefit in well-selected patients with PCL.


HER2-Overexpressing/Amplified Breast Cancer as a Testing Ground for Antibody-Drug Conjugate Drug Development in Solid Tumors.
Efficacy data from the KATHERINE clinical trial, comparing the HER2-directed antibody-drug conjugate (ADC) ado-trastuzumab emtansine (T-DM1) to trastuzumab in patients with early-stage HER2-amplified/overexpressing breast cancer with residual disease after neoadjuvant therapy, demonstrates superiority of T-DM1 (HR for invasive disease or death, 0.50; P < 0.001). This establishes foundational precedent for ADCs as effective therapy for treatment of subclinical micrometastasis in an adjuvant (or post-neoadjuvant) early-stage solid tumor setting. Despite this achievement, general principles from proposed systems pharmacokinetic modeling for intracellular processing of ADCs indicate potential shortcomings of T-DM1: (i) C max limited by toxicities; (ii) slow internalization rate; (iii) resistance mechanisms due to defects in intracellular trafficking [loss of lysosomal transporter solute carrier family 46 member 3, (SLC46A3)], and increased expression of drug transporters MDR1 and MRP1; and (iv) lack of payload bystander effects limiting utility in tumors with heterogeneous HER2 expression. These handicaps may explain the inferiority of T-DM1-based therapy in the neoadjuvant and first-line metastatic HER2+ breast cancer settings, and lack of superiority to chemotherapy in HER2+ advanced gastric cancer. In this review, we discuss how each of these limitations is being addressed by manipulating internalization and trafficking using HER2:HER2 bispecific or biparatopic antibody backbones, using site-specific, fixed DAR conjugation chemistry, and payload swapping to exploit alternative intracellular targets and to promote bystander effects. Newer HER2-directed ADCs have impressive clinical activity even against tumors with lower levels of HER2 receptor expression. Finally, we highlight ongoing clinical efforts to combine HER2 ADCs with other treatment modalities, including chemotherapy, molecularly targeted therapies, and immunotherapy.


Lipopolysaccharide-stimulated PGE2 release from human monocytes. Comparison of lipopolysaccharides prepared from suspected periodontal pathogens.
Lipopolysaccharides (LPS) prepared from the suspected periodontal pathogens Actinobacillus actinomycetemcomitans (A. a.), Bacteroides gingivalis, B. intermedius and Wolinella recta were compared to Salmonella typhimurium LPS for their capacity to stimulate prostaglandin E2 (PGE2) release from human monocytes. Counterflow isolated monocytes were cultured with control medium or media containing 10 micrograms/ml LPS. Media were then exchanged every 24 hours for a total of 72 hours. Salmonella and Wolinella LPS preparations demonstrated seven-fold greater PGE2 release than B. gingivalis and two-fold greater than A. a. and B. intermedius. PGE2 release was found to decrease over time with all LPS preparations except Wolinella. The potency of the LPS preparations is tentatively ranked as follows: Wolinella greater than or equal to Salmonella greater than A. a. greater than B. intermedius greater than or equal to B. gingivalis. These findings demonstrate that LPS preparations from suspected periodontal pathogens are capable of stimulating PGE2 release from human monocytes. The high potency and prolonged stimulation of PGE2 release with Wolinella LPS suggests unusual toxic properties that may exert a greater influence in the pathogenesis of destructive periodontal diseases.


Effect of nest temperature on egg-brooding dynamics in Children's pythons.
Parental care meets several critical needs of developing offspring. In particular, egg brooding in pythons is an exceptional model for examining environmental influences on specific parental behaviors because brooding behaviors are dynamic yet simple and modulate embryonic temperature, respiration, and water balance. We used captive Children's pythons (Antaresia childreni) to assess their facultative endothermic capability, the influence on egg-brooding behaviors of the gradient between the nest temperature (T(nest)) and clutch temperature (T(clutch)), and the effect of these behaviors on the developmental micro-environment. We monitored maternal egg-brooding behavior, rates of brooding unit (i.e., female and associated clutch) respiratory gas exchange, T(nest), T(clutch), and intra-clutch oxygen tension (PO(2)clutch) during acute changes among four incubation temperature conditions: constant preferred temperature (31.5 degrees C); 'cooling' (T(nest)<T(clutch)); constant cool temperature (25.6 degrees C); and 'warming' (T(nest)>T(clutch)). We demonstrated that A. childreni are not facultatively endothermic because brooding unit temperature coefficient (Q(10)) for V(O2) and V(CO2) was similar to other ectothermic boid snakes (1.9-5.7) and T(clutch) conformed to T(nest) at the constant, cool temperature treatment. Females coiled tightly around eggs more often during cooling than during warming. Further, the amount of time that females spent tightly coiled during warming significantly affected the T(nest)-T(clutch) gradient. Together these results indicate that, although female A. childreni are not facultatively endothermic, they are capable of assessing the T(nest)-T(clutch) gradient and making behavioral adjustments to enhance the thermal micro-environment of their developing offspring.


MRI detection of tissue pathology beyond atrophy in Alzheimer's disease: introducing T2-VBM.
Voxel-based morphometry (VBM) of T1-weighted magnetic resonance (MR) images has been widely used to identify regional atrophy in neurodegenerative conditions such as Alzheimer's disease (AD). In theory, however, T2-weighting should be more sensitive to tissue pathology, though until recently, volumetric T2-weighted images were unavailable. We tested the hypothesis that T2-VBM would be more sensitive to grey matter pathology in AD than T1-VBM using the recently-developed SPACE acquisition, which provides true-3D, high-resolution T2-weighted images. This was contrasted to conventional T1-weighted MPRAGE images acquired at the same session and resolution. All of the atrophic regions identified with T1-VBM were also identified with T2-VBM. Additional abnormalities were, however, identified with T2-VBM and the distribution of these bore a striking resemblance to the distribution of amyloid plaque deposition in AD, suggesting that T2-VBM detects signal changes due to histopathology over and above those attributable to atrophy. In keeping with this hypothesis, the relevant statistical tests demonstrated that the difference in sensitivity was caused by an apparent change in T2-weighted signal intensity that was not present in T1-weighted images. These results suggest that T2-VBM has the potential to advance VBM beyond atrophy detection to more expansive applications in tissue pathology mapping.


[Prevalence of overweight and obesity in Swiss adolescents].
Overweight and obesity are the most important modulators in the development of chronic diseases. Body weight status in adulthood is often already determined in childhood and adolescence. In this study we evaluate possible threats and the prevalence of overweight and obesity among students of the Kantonsschule Burggraben in St.Gallen. Different anthropometric parameters (e.g. size, weight, circumference of waist and hip, body composition by BIA) were assessed among 552 students and different life-style factors as well as risk factors were assessed with a self-administered questionnaire. Using the IOTF-definitions between 6.12-8.18% of students were overweight and 1.08-2.23% were obese. Male students with the highest media consumption (MC) (around 4 h/d) had a higher BMI (kg/m2) than students with lower MC (around 2 h/d) or the lowest level of MC (approximately 1 h/d) (22.83 +/- 0.46 kg/m2 vs. 21.51 +/- 0.21 kg/m2 and 21.60 +/- 0.39 kg/m2; ANOVA p = 0.008). Similarly body composition was affected: students with the highest media consumption had the highest amount of body-fat mass: 12.69 +/- 0.66% vs. 11.05 +/- 0.34% and 10.69 +/- 0.62% (ANOVA p = 0.026) compared to medium and low MC, respectively. There was no correlation between the BMI, the body fat mass and the MC among female students. According to the level of activity male students with a high level showed a significant lower BMI than students with a lower level of physical activity (21.70 +/- 0.21 kg/m2 vs. 22.05 +/- 0.33 kg/m2; p = 0.016). The body fat mass was also lower for higher levels of physical activity (9.57 +/- 0.65% vs. 12.69 +/- 0.86%). Among female students the level of physical activity modulated the body fat mass (high level 20.66 +/- 0.52% vs. low level 23.53 +/- 0.58; p = 0.005) but had no effect on the BMI. Risk factors for obesity such as physical activity patterns are established in childhood and early adolescence. Preventive strategies have to be implemented as early as possible.


Nonhuman Primates: A Vital Model for Basic and Applied Research on Female Reproduction, Prenatal Development, and Women's Health.
The comparative biology of reproduction and development in mammalian species is remarkable. Hence, because of similarities in environmental and neuroendocrine control of the reproductive axis, the cyclic function of the ovary and reproductive tract, establishment and control of the maternal-fetal-placental unit during pregnancy, and reproductive aging from puberty through menopause, nonhuman primates (NHPs) are valuable models for research related to women's reproductive health and its disorders. This chapter provides examples of research over the past 10+ years using Old World monkeys (notably macaque species), baboons, and to a lesser extent New World monkeys (especially marmosets) that contributed to our understanding of the etiology and therapies or prevention of: (1) ovarian disorders, e.g., polycystic ovary syndrome, mitochondrial DNA-based diseases from the oocyte; (2) uterine disorders, for example, endometriosis and uterine transplantation; and (3) pregnancy disorders, for example, preterm labor and delivery, environmental factors. Also, emerging opportunities such as viral (e.g., Zika) induced fetal defects and germline genomic editing to generate valuable primate models of human diseases (e.g., Huntington and muscular dystrophy) are addressed. Although the high costs, specialized resources, and ethical debate challenge the use of primates in biomedical research, their inclusion in fertility and infertility research is vital for continued improvements in women's reproductive health.


Noninterferometric quantitative phase imaging with soft x rays.
We demonstrate quantitative noninterferometric x-ray phase-amplitude measurement. We present results from two experimental geometries. The first geometry uses x rays diverging from a point source to produce high-resolution holograms of submicrometer-sized objects. The measured phase of the projected image agrees with the geometrically determined phase to within +/-7%. The second geometry uses a direct imaging microscope setup that allows the formation of a magnified image with a zone-plate lens. Here a direct measure of the object phase is made and agrees with that of the magnified object to better than +/-10%. In both cases the accuracy of the phase is limited by the pixel resolution.


Recombinationless meiosis in Saccharomyces cerevisiae.
We have utilized the single equational meiotic division conferred by the spo13-1 mutation of Saccharomyces cerevisiae (S. Klapholtz and R. E. Esposito, Genetics 96:589-611, 1980) as a technique to study the genetic control of meiotic recombination and to analyze the meiotic effects of several radiation-sensitive mutations (rad6-1, rad50-1, and rad52-1) which have been reported to reduce meiotic recombination (Game et al., Genetics 94:51-68, 1980); Prakash et al., Genetics 94:31-50, 1980). The spo13-1 mutation eliminates the meiosis I reductional segregation, but does not significantly affect other meiotic events (including recombination). Because of the unique meiosis it confers, the spo13-1 mutation provides an opportunity to recover viable meiotic products in a Rec- background. In contrast to the single rad50-1 mutant, we found that the double rad50-1 spo13-1 mutant produced viable ascospores after meiosis and sporulation. These spores were nonrecombinant: meiotic crossing-over was reduced at least 150-fold, and no increase in meiotic gene conversion was observed over mitotic background levels. The rad50-1 mutation did not, however, confer a Rec- phenotype in mitosis; rather, it increased both spontaneous crossing-over and gene conversion. The spore inviability conferred by the single rad6-1 and rad52-1 mutations was not eliminated by the presence of the spo13-1 mutation. Thus, only the rad50 gene has been unambiguously identified by analysis of viable meiotic ascospores as a component of the meiotic recombination system.


Effects of xylazine and ketamine on the acoustic reflex and brain stem auditory-evoked response in the cat.
The acoustic reflex (AR) and brain stem auditory-evoked response (BAER) were recorded in adult cats 5 minutes after IM administration of xylazine (1 mg/kg) and after IM administration of ketamine (10 mg/kg). Ipsilateral and contralateral AR were recorded at 10 and 20 dB above acoustic reflex threshold 5 minutes after xylazine administration and 5 and 35 minutes after ketamine administration. Monaural BAER were recorded 5 minutes after xylazine and 5 and 35 minutes after ketamine, using stimulus intensities of 90-, 80-, and 70-dB hearing level (HL). Additional BAER were recorded at 10, 15, and 25 minutes after ketamine, using the 90-dB HL stimulus. Pre- and postinjection comparisons were made for threshold, latency, and amplitude of the AR and for latency and amplitude of waves I through VI of the BAER. At both stimulus intensities before and after ketamine administration threshold for the ipsilateral reflex was significantly lower (P greater than 0.05) than for the contralateral reflex. The threshold, latency, and amplitude of the AR were unaffected (P greater than 0.05) by the injection of ketamine after xylazine. The amplitude of BAER waves was not affected (P greater than 0.05) by ketamine after xylazine for each of the 3 stimulus intensities. Latency of the 90-dB HL-evoked response was increased (P less than or equal to 0.05) for waves III/IV at 5 and 35 minutes after ketamine, and for wave V at each of the postinjection times, except at postinjection minute 15.(ABSTRACT TRUNCATED AT 250 WORDS)


[Malignant pleural mesothelioma. Value of 67Ga scintigraphy compared to computerized tomography].
In 34 patients with suspected malignant pleural mesothelioma the results of computed tomography are compared with the findings of 67Ga-scintigraphy. The differential diagnosis of 14 pleural mesotheliomas, 7 pleural carcinoses, 10 inflammatory and 3 other pleural diseases is performed more accurately by CT than by scintigraphy. 67Ga uptake depends on the thickness of inflammatory as well as malignant lesions. Thus, numerous pleural processes that can be localised by CT escape scintigraphic detection. CT is indicated if there is clinical and radiological suspicion of pleural mesothelioma; in that case, there is hardly any indication for 67Ga scintigraphy.


Community-based distribution of family planning services in Saradidi, Kenya.
Community-based distribution (CBD) of family planning services was initiated in 1980 in Saradidi, Kenya, as part of a community development effort. Family planning information and services in each village were provided by volunteer health helpers (VHH's) chosen and supported by the people in each village. The initial examination and supply of commodities was provided at a community clinic. Less than 1% of women 15 to 49 years of age used a family planning method before CBD was initiated. In 1983, 31 (17.3%) of 179 randomly selected currently married women and 26 (52.0%) of 50 currently married VHH's reported having used a family planning method; 38 (66.7%) were still using a method at the time of the survey. Family planning use increased with age and education. Women who used family planning had higher parity, were less likely to want more children and had had a longer time since the last delivery. From 1980 to 1983, 732 persons (including 121 men) were seen at the Saradidi clinic requesting family planning services; 17.2% were referred from the VHH's. About one-third of clients referred from VHH's to the clinic for examination and commodities actually came. Allowing VHH's to carry out the initial examination and provide the first supply of commodities to the acceptors might have significantly increased the rate of family planning use. The findings demonstrate an increased use of family planning services in Saradidi following the inception of CBD.


Consistency analysis of microRNA-arm expression reveals microRNA-369-5p/3p as tumor suppressors in gastric cancer.
The 5p and 3p arms of microRNA (miRNA) are typically generated from the same precursor, and one arm influences protein output, while the other has a short half-life. However, a few miR-5p/3p pairs have been reported to co-exist in cancer cells. Here, we performed a genome-wide analysis of miRNA expression in gastric cancer (GC) cells to systematically investigate the co-expression profile of miR-5p/3p in gastric tumorigenesis. We discovered that only 41 miR-5p/3p pairs out of 1749 analyzed miRNA were co-expressed. Specifically, abnormal expression of miR-369-5p and miR-369-3p was correlated with GC progression. Importantly, both in vitro and in vivo assays revealed that miR-369-5p and miR-369-3p exhibited tumor-suppressive roles by regulating jun proto-oncogene and v-akt murine thymoma viral oncogene homolog 1 function in GC cells, respectively. Moreover, we observed that miR-369 was inactivated in GC tissues due to DNA methylation. We also showed that inhibition of miR-369-5p/3p attenuated the effect of azacitidine (AZA) treatment on suppressing cell growth and invasion. These results suggest that the therapeutic efficacy of AZA in GC is at least partly attributable to miR-369 activation. Overall, our findings provide convincing evidence that both the 5p and 3p arms of miRNA co-expressed in GC and DNA methylation-induced miR-369 signaling contribute to GC progression.


Bacteremia with Streptococcus pneumoniae. Implications for therapy and prevention. Franklin County Pneumonia Study Group.
To determine the incidence and mortality rates of patients with bacteremic infections with Streptococcus pneumoniae, and to determine the serotypes and antimicrobial susceptibilities of the pneumococcal isolates. Prospective case ascertainment and procurement of S pneumoniae isolates between January 1991 and April 1994. Ten adult care hospitals in Franklin County, Ohio. Patients (N = 590) in whom S pneumoniae was isolated from blood cultures. Demographic data from patients with pneumococcal bacteremia were obtained by chart review. Pneumococcal serotyping and antimicrobial susceptibility testing were performed on 499 bacteremic isolates. Among residents of Franklin County, the annual incidence of pneumococcal bacteremia was higher in patients at least 65 years old (83.0 per 100,000 population) compared with younger adults (9.6 per 100,000 population) (odds ratio [OR], 8.74; 95% confidence interval [CI], 7.19 to 10.62) and more common among African Americans than whites (OR, 1.36; 95% CI, 1.06 to 1.75). The relative risk of pneumococcal bacteremia among persons infected with the human immunodeficiency virus was 41.8 times (CI, 19.0 to 92.0) that of county residents 18 to 64 years of age. The overall mortality rate was 19% and was age-dependent, reaching 38% in patients at least 85 years old. The distribution of pneumococcal serotypes causing bacteremia was remarkably consistent over time. The incidence of drug-resistant strains increased during the study period; by 1994 14% were resistant to penicillin, 12% to ceftazidime, and 24% to trimethoprim-sulfamethoxazole. The resistant strains included several serotypes of S pneumoniae. Most serotypes (89.8%) of S pneumoniae causing bacteremia are contained in the pneumococcal vaccine. Increased use of pneumococcal vaccine and recognition of antimicrobial resistance patterns may assist physicians in treating patients with S pneumoniae bacteremia. Educational programs to discourage unnecessary antimicrobial drug use should be developed for patients and physicians.


Diabetic charcot neuroarthropathy of the foot and ankle with osteomyelitis.
One of the most devastating foot and/or ankle complications in the diabetic population with peripheral neuropathy is the presence of Charcot neuroarthropathy (CN). In recent years, diabetic limb salvage has been attempted more frequently as opposed to major lower extremity amputation for CN of the foot and ankle with ulceration and/or deep infection. Treatment strategies for osteomyelitis in the diabetic population have evolved. This article reviews some of the most common surgical strategies recommended for the diabetic patient with CN of the foot and/or ankle and concomitant osteomyelitis.


Principles of state health reform: many states not willing to wait for federal reform.
Despite the simplicity of the basic objectives of health care reform-- greater access at manageable cost, these goals have not yet been achieved at either the federal or state level. One explanation may be that the American people are not willing to make the sacrifices that are probably necessary to achieve universal access to health care: increased taxes or redirection of governmental expenditures, limitation of choice in providers, and perhaps some form of rationing (which in fact already exists, by limiting access of the uninsured and some of the poor). What, then, are the prospects for meaningful national health care reform in the near future? While the answer to this question remains unclear, there is no doubt that providers across the country are likely to face an unprecedented array of state health care initiatives over the next few years, whether or not federal legislation is enacted. To prepare for this upcoming legislative activity, providers must remain aware of state legislative activity as it evolves.


[Morphology of the endocrine portion of the reptile pancreas].
The endocrinous part of the pancreatic gland of reptiles Lacerta agilis, Agama sanguinolenta, Varanus griseus, Testudo horsfieldi and Clemmus Caspica is formed by A-, B- and D-cells. The main form of its structural organization is pancreatic islands. In adition to them, remnants of the "external epithelium" are found in Varanus griseus and A-cells disposed outside of the pancreatic islands in Testudo horsfieldi and Clemmys caspica. As compared with amphibia, reptiles have a better developed counterinsular component (A-cells) of the endocrinous part of the pancreas. The acino-island cells of "A" and "D" types are also found in the pancreas of reptiles, "A"-type cells being predominant among them.


[Meta-analysis of randomized controlled trials comparing procedure for prolapse and hemorrhoids with Milligan-Morgan hemorrhoidectomy in the treatment of prolapsed hemorrhoids].
To compare the safety and efficacy of procedure for prolapse and hemorrhoids (PPH) with Milligan-Morgan hemorrhoidectomy (MMH) in the treatment of prolapsed hemorrhoids. All the randomized controlled trials (RCT) comparing PPH with MMH in the treatment of prolapsed hemorrhoids published between January 1998 and January 2015 were retrieved from PubMed, Embase, Cochrane Library, CBM, CNKI, Wangfang, VIP databases. Hand search was applied in Chinese Journal of Gastrointestinal Surgery, Chinese Journal of Coloproctology and Journal of Colorectal and Anal Surgery from the library of Chengdu University of Traditional Chinese Medicine. Associated reference documents in enrolled trials were reviewed. The methodological quality of enrolled trials was evaluated according to the Cochrane handbook for systematic reviews of interventions. Meta-analysis was performed using RevMan 5.3 software. Sixteen RCTs recruiting 1411 patients were identified. Among them, 702 patients underwent PPH, and other 709 patients MMH. Meta-analysis showed that as compared to MMH, PPH had shorter operating time(WMD=-12.34, 95% CI:-17.87 to -6.80, P=0.000], shorter hospital stay (WMD=-1.48, 95% CI: -1.81 to -1.14, P=0.000) and shorter time to recover normal activity (WMD=-14.11, 95% CI: -24.51 to -3.70, P=0.008). Patients in PPH group experienced less pain at postoperative 24 h, postoperative 1 week and the first postoperative bowel movement (all P<0.01). PPH was more ascendant in terms of the requirement for analgesics(P<0.01). PPH group had higher ratio of wound-healing 2 weeks after surgery (RR=0.19, 95% CI: 0.07 to 0.51, P=0.001), lower ratio of postoperative anal stenosis (RR=0.39, 95% CI: 0.15 to 0.99, P=0.050) and lower ratio of anal incontinence (RR=0.62, 95% CI: 0.38 to 1.01, P=0.050), but higher ratio of recurrent disease after 1 year (RR=2.54, 95% CI: 1.21 to 5.31, P=0.010). No significant differences in ratios of postoperative bleeding, urinary retention, and postoperative morbidity of complication were found(all P>0.05). Satisfaction degree of patients in PPH group was better than that in MMH group (OR=2.36, 95% CI:1.36 to 4.07, P=0.002). Procedure for prolapse and hemorrhoids offers some short-term benefits over Milligan-Morgan hemorrhoidectomy, but is associated with a higher rate of recurrent disease.


Continued transplant immunosuppression may prolong survival after return to peritoneal dialysis: results of a decision analysis.
Patient survival is prolonged in those with residual renal function. The risks of opportunistic infection and malignancy militate against a widespread policy of maintaining immunosuppression in those returning to peritoneal dialysis after a failed allograft. We set out to test the hypothesis that patients returning to peritoneal dialysis after a failed transplant benefit from continued immunosuppression. A decision analytic model comparing the use of immunosuppression after transplant failure and return to peritoneal dialysis with immunosuppressive withdrawal. Two assumptions were made: that the survival benefit reported in patients with a transplant kidney was the same as that expected from a native kidney with a similar glomerular filtration rate (GFR) and that the risks of carcinoma and opportunistic infections were equal to that of the general population if immunosuppressive therapy was discontinued. Probabilities were obtained from the literature, from an administrative database, and from the rate of GFR decline seen in our own unit in patients returning to dialysis who had been discontinued from their transplant immunosuppressive medication. Life expectancy was prolonged from 5.3 years to 5.8 years when immunosuppression was continued in patients who returned to dialysis after chronic allograft failure. A higher survival benefit was seen at higher levels of additional GFR. A benefit from continued immunosuppressive therapy was seen for all values of additional GFR greater than 15 L/wk. All dialysis and renal clearances have been expressed as L/wk. The conversion factors for conversion to mL/s and mL/min are 605 and 10.1. The data suggest that there may be a survival advantage in maintaining patients on long-term immunosuppressive function even after they return to peritoneal dialysis after allograft failure.


Interaction of Zn2+ with guinea-pig C5 convertase and guinea-pig C5.
A comparison of two methods of C5 activation, the standard method (EAC14 + C2, C3, C5, C6, and C7) and the washed-cell intermediate method (EAC1423 + C5 and washed), demonstrated that formation of hemolytically competent SAC14235 was reduced in the washed-cell method. Addition of Zn2+ in this method increased the formation of competent SAC14235 to the approximate level of the standard method. The optimum concn range of Zn2+ was 0.006-0.025 mM. In the standard method Zn2+ had no significant enhancing effect on the formation of competent SAC14235. Zn2+ had a greater affinity for EAC1423 than for fluid-phase C5 when reacted with each separately. Maximum enhancement, however, was obtained when Zn2+ was present during the reaction of C5 with EAC1423. The effect of Zn2+ on C5 activity was not related to the stabilizing property of C6 on cell-bound C5. In the washed-cell method there was an inverse relationship between the concn of C2 or C3 and the concn of C5 required to generate one competent SAC14235/cell. Over a 10-fold range of C2 or C3 concn there was an approximate four-fold increase in the number of competent SAC14235/cell formed in the presence of Zn2+. Zn2+ enhances formation of competent SAC14235 on cells that have a limited ability to activate C5 even though the C2 and C3 on the convertase are present in excess.


Multi-annual performance evaluation of laboratories in post-mortem diagnosis of animal rabies: Which techniques lead to the most reliable results in practice?
Rabies diagnosis proficiency tests on animal specimens using four techniques (FAT, RTCIT, conventional RT-PCR and real-time RT-PCR) were organised over 10 years (2009-2019). Seventy-three laboratories, of which 59% were from Europe, took part. As the panels were prepared with experimentally-infected samples, the error rate of laboratories on positive and negative samples was accurately estimated. Based on fitted values produced by mixed modelling including the variable "laboratory" as a random variable to take into account the longitudinal design of our dataset, the technique that provided the most concordant results was conventional RT-PCR (99.3%; 95% CI 99.0-99.6), closely followed by FAT (99.1%; 95% CI 98.7-99.4), real-time RT-PCR (98.7%; 95% CI 98.1-99.3) and then RTCIT (96.8%; 95% CI 95.8-97.7). We also found that conventional RT-PCR provided a better diagnostic sensitivity level (99.3% ±4.4%) than FAT (98.7% ±1.6%), real-time RT-PCR (97.9% ±0.8%) and RTCIT (95.3% ±5.1%). Regarding diagnostic specificity, RTCIT was the most specific technique (96.4% ±3.9%) followed closely by FAT (95.6% ±3.8%), real-time RT-PCR (95.0% ±1.8%) and conventional RT-PCR (92.9% ±0.5%). Due to multiple testing of the samples with different techniques, the overall diagnostic conclusion was also evaluated, and found to reach an inter-laboratory concordance level of 99.3%. The concordance for diagnostic sensitivity was 99.6% ±2.0% and for diagnostic specificity, 98.0% ±8.5%. Molecular biology techniques were, however, found to be less specific than expected. The potential reasons for such findings are discussed herein. The regular organisation of performance tests has contributed to an increase in the performance of participating laboratories over time, demonstrating the benefits of such testing. Maintaining a high-quality rabies diagnosis capability on a global scale is key to achieving the goal of eliminating dog-mediated human rabies deaths. The regular organisation of exercises on each continent using selected local strains to be tested according to the local epidemiological situation is one factor that could help increase reliable diagnosis worldwide. Rabies diagnosis capabilities could indeed be enhanced by providing adequate and sustainable proficiency testing on a large scale and in the long term.


Effector molecules released by Th1 but not Th17 cells drive an M1 response in microglia.
Microglia act as sensors of inflammation in the central nervous system (CNS) and respond to many stimuli. Other key players in neuroinflammatory diseases are CD4+ T helper cell (Th) subsets that characteristically secrete IFN-γ (Th1) or IL-17 (Th17). However, the potential of a distinct cytokine milieu generated by these effector T cell subsets to modulate microglial phenotype and function is poorly understood. We therefore investigated the ability of factors secreted by Th1 and Th17 cells to induce microglial activation. In vitro experiments wherein microglia were cultured in the presence of supernatants derived from polarized Th1 or Th17 cultures, revealed that Th1-associated factors could directly activate and trigger a proinflammatory M1-type gene expression profile in microglia that was cell-cell contact independent, whereas Th17 cells or its associated factors did not have any direct influence on microglia. To assess the effects of the key Th17 effector cytokine IL-17A in vivo we used transgenic mice in which IL-17A is specifically expressed in astrocytes. Flow cytometric and histological analysis revealed only subtle changes in the phenotype of microglia suggesting only minimal effects of constitutively produced IL-17A on microglia in vivo. Neither IL-23 signaling nor addition of GM-CSF, a recently described effector molecule of Th17 cells, changed the incapacity of Th17 cells to activate microglia. These findings demonstrate a potent effect of Th1 cells on microglia, however, the mechanism of how Th17 cells achieve their effect in CNS inflammation remains unclear.


Nuclear S100A7 is associated with poor prognosis in head and neck cancer.
Tissue proteomic analysis of head and neck squamous cell carcinoma (HNSCC) and normal oral mucosa using iTRAQ (isobaric tag for relative and absolute quantitation) labeling and liquid chromatography-mass spectrometry, led to the identification of a panel of biomarkers including S100A7. In the multi-step process of head and neck tumorigenesis, the presence of dysplastic areas in the epithelium is proposed to be associated with a likely progression to cancer; however there are no established biomarkers to predict their potential of malignant transformation. This study aimed to determine the clinical significance of S100A7 overexpression in HNSCC. Immunohistochemical analysis of S100A7 expression in HNSCC (100 cases), oral lesions (166 cases) and 100 histologically normal tissues was carried out and correlated with clinicopathological parameters and disease prognosis over 7 years for HNSCC patients. Overexpression of S100A7 protein was significant in oral lesions (squamous cell hyperplasia/dysplasia) and sustained in HNSCC in comparison with oral normal mucosa (p(trend)<0.001). Significant increase in nuclear S100A7 was observed in HNSCC as compared to dysplastic lesions (p = 0.005) and associated with well differentiated squamous cell carcinoma (p = 0.031). Notably, nuclear accumulation of S100A7 also emerged as an independent predictor of reduced disease free survival (p = 0.006, Hazard ratio (HR = 7.6), 95% CI = 1.3-5.1) in multivariate analysis underscoring its relevance as a poor prognosticator of HNSCC patients. Our study demonstrated nuclear accumulation of S100A7 may serve as predictor of poor prognosis in HNSCC patients. Further, increased nuclear accumulation of S100A7 in HNSCC as compared to dysplastic lesions warrants a large-scale longitudinal study of patients with dysplasia to evaluate its potential as a determinant of increased risk of transformation of oral premalignant lesions.


Role of the terminal domains in sodium channel localization.
Voltage-gated sodium channels are membrane proteins that initiate action potentials in neurons following membrane depolarization. Members of this family show differential distribution at the subcellular level. The mechanisms underlying the targeting of these isoforms are not understood. However, their specificity is important because the isoforms can change the excitability of the membrane due to differences in their electrophysiological properties. In this study, chimeras generated between Na(V)1.2 and Na(V)1.6 were used to test channel domains for sequence that would allow Na(V)1.2 to localize to unmyelinated axons when Na(V)1.6 could not. We show that the N-terminal 202 amino acids of the Na(V)1.2 channel can mediate membrane domain-specific sorting in polarized epithelial cells and are necessary but not sufficient for localizing the isoform to the axons of cultured neurons. The domain-sorting signal is in the region between amino acids 110-202 of the Na(V)1.2 channel. The C-terminal 451 amino acids of Na(V)1.2 likely contain determinants that interact with neuron-specific factors to direct Na(V)1.2 to the axon.


iTRAQ-based Quantitative Proteomics Analysis Identifies Host Pathways Modulated during Toxoplasma gondii Infection in Swine.
Toxoplasma gondii is a leading cause of foodborne illness and consumption of undercooked pig meat is a major risk factor for acquiring toxoplasmosis, which causes a substantial burden on society. Here, we used isobaric tags for relative and absolute quantification (iTRAQ) labelling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify cellular proteins and pathways altered during T. gondii infection in pigs. We also used parallel reaction monitoring-based LC-MS/MS to verify the levels of protein expression of infected spleens and mesenteric lymph nodes (MLNs). At 6 days post-infection (dpi), 156, 391, 170, 292, and 200 differentially expressed proteins (DEPs) were detected in the brain, liver, lung, MLNs and spleen, respectively. At 18 dpi, 339, 351, 483, 388, and 303 DEPs were detected in the brain, liver, lung, MLNs and spleen, respectively. Although proteins involved in immune responses were upregulated in all infected tissues, protein expression signature in infected livers was dominated by downregulation of the metabolic processes. By weighted gene co-expression network analysis, we could further show that all proteins were clustered into 25 co-expression modules and that the pink module significantly correlated with the infection status. We also identified 163 potential anti-T. gondii proteins (PATPs) and provided evidence that two PATPs (HSP70.2 and PDIA3) can reduce T. gondii burden in porcine macrophages in vitro. This comprehensive proteomics analysis reveals new facets in the pathogenesis of T. gondii infection and identifies key proteins that may contribute to the pig's defense against this infection.


The fecal microbiota of Thai school-aged children associated with demographic factors and diet.
Birth delivery method and breastfeeding practices contribute to microbiota colonization. Other factors including diet and demographic factors structure the gut microbiome assembly and diversity through childhood development. The exploration of these factors, especially in Southeast Asian children, remains limited. We investigated the fecal microbiota of 127 school-aged children in Thailand using quantitative PCR (qPCR) to assess the influence of diet and demographic factors on the gut microbiota. Multivariate analysis (multiple factor analysis (MFA) and Partial Least Squares Discriminant Analysis (PLS-DA)) were used to link particular gut microbes to diet and demographic factors. Diet and demographic factors were associated with variation among gut microbiota. The abundance of Gammaproteobacteria increased in children with infrequent intake of high fat foods. Obese children possessed a lower level of Firmicutes and Ruminococcus. Bifidobacterium was enriched in pre-teen aged children and detected at lower levels among formula-fed children. Prevotella was more abundant in children who were delivered vaginally. While ethnicity explained a small amount of variation in the gut microbiota, it nonetheless was found to be significantly associated with microbiome composition. Exogenous and demographic factors associate with, and possibly drive, the assembly of the gut microbiome of an understudied population of school-aged children in Thailand.


The effect of heavy maternal alcohol intake on amniotic fluid phospholipids in late pregnancy.
The reported effects of alcohol on lung and brain phospholipid metabolism in adult animals have been numerous, but its effect on the fetal lung has not been examined. We investigated the effect of heavy maternal ethanol intake on fetal pulmonary maturation, as measured by the amniotic fluid phospholipids, lecithin and phosphatidylglycerol (PG). Mothers were screened at their first prenatal visit, and detailed drinking histories were taken. Amniotic fluid phospholipids were determined later in pregnancy for clinical purposes, and mothers with positive scores who admitted to drinking during the third trimester were compared to a control group. Amniotic fluid from mothers whose fetuses were exposed to ethanol late in pregnancy did not show the expected increase in lecithin/sphingomyelin and PG related to gestational age that was observed in nondrinking mothers. Canonical correlation of gestational age, late pregnancy drinking, and chronic alcohol problems with amniotic fluid L/S and PG jointly revealed that both gestational age and maternal drinking were significant determinants of fetal lung maturity (r = .39, P less than .05), with active maternal drinking being about one-third as important a determinant as gestational age. These findings could be explained by the known damage-inducing effect of ethanol on alveolar cell membranes or by the effects of ethanol on phospholipid metabolism.


Accelerated Idioventricular Rhythm in a Child With Status Asthmaticus.
Acute asthma exacerbations occur relatively frequently in children. We present the case of a 4-year-old boy who was admitted to our hospital in status asthmaticus and found to have a wide complex rhythm while being treated with inhaled albuterol and intravenous methylprednisolone. This rhythm was diagnosed as accelerated idioventricular rhythm (AIVR), which carries a benign prognosis. It resolved as the medications used to treat his asthma exacerbation were weaned. There was no ventricular ectopy seen on a 24-hour Holter monitor performed 3 months after his hospitalization, suggesting that the AIVR was related to the medications the patient was receiving at the time. This case suggests that albuterol may be a risk factor for the development of AIVR and highlights the importance of recognizing this rhythm to avoid unnecessary and potentially harmful therapies.


Recent developments of genomic research in soybean.
Soybean is an important cash crop with unique and important traits such as the high seed protein and oil contents, and the ability to perform symbiotic nitrogen fixation. A reference genome of cultivated soybeans was established in 2010, followed by whole-genome re-sequencing of wild and cultivated soybean accessions. These efforts revealed unique features of the soybean genome and helped to understand its evolution. Mapping of variations between wild and cultivated soybean genomes were performed. These genomic variations may be related to the process of domestication and human selection. Wild soybean germplasms exhibited high genomic diversity and hence may be an important source of novel genes/alleles. Accumulation of genomic data will help to refine genetic maps and expedite the identification of functional genes. In this review, we summarize the major findings from the whole-genome sequencing projects and discuss the possible impacts on soybean researches and breeding programs. Some emerging areas such as transcriptomic and epigenomic studies will be introduced. In addition, we also tabulated some useful bioinformatics tools that will help the mining of the soybean genomic data.


Healthcare Staffs' Information Security Practices Towards Mitigating Data Breaches: A Literature Survey.
The purpose of this study was to understand healthcare staffs' information security (IS) practices towards mitigating data breaches. A literature survey was conducted to understand the state-of-the-art methods, tools, evaluation techniques and the challenges to their implementation. The results would be used for emperical studies in a hospital setting in Norway on Healthcare Security Practice Analysis, Modeling and Incentivization (HSPAMI). Human Aspect of Information Security Questionnaire was identified as robust and comprehensive tool for gathering staff security practices. Integrated theories was being adopted to form a comprehensive staffs' characteristics. A mixed-method for evaluating the theories was also identified to be the best strategy.


PepFect14 mediates the delivery of mRNA into human primary keratinocytes and in vivo.
mRNA-based vaccines and candidate therapeutics have great potential in various medical fields. For the delivery of mRNA into target cells and tissues, lipid formulations are often employed. However, this approach could cause the activation of immune responses, making it unsuitable for the treatment of inflammatory conditions. Therefore, alternative delivery systems are highly demanded. In this study, we evaluated the transport efficiency and characteristics of cell-penetrating peptide PepFect14 (PF14) and mRNA nanoparticles in the presence of different additives. Our results show that all PF14-mRNA formulations entered cultured cells, while calcium chloride enhanced the transport and production of the encoded protein in HeLa and HaCaT cell lines, and polysorbate 80 did so in primary human keratinocytes. All formulations had similar physical properties and did not remarkably affect cell viability. By selectively blocking endocytosis pathways, we show that PF14-mRNA nanoparticles primarily entered HeLa cells via macropinocytosis and HaCaT cells via both macropinocytosis and clathrin-mediated endocytosis, while none of the blockers significantly affected the delivery into primary keratinocytes. Finally, subcutaneous injection of PF14-mRNA nanoparticles before inducing mouse irritant contact dermatitis resulted in the expression of a reporter protein without provoking harmful immune responses in the skin. Together, our findings suggest that PF14-mRNA nanoparticles have the potential for developing mRNA-based therapeutics for treating inflammatory skin conditions.


Asymmetric Synthesis of 2'-C-Methyl-4'-selenonucleosides as Anti-Hepatitis C Virus Agents.
In search of a new template for anti-hepatitis C virus (HCV) agents, we designed and synthesized the 2'-C-methyl-4'-selenopyrimidine and -purine nucleosides and their phosphoramidate prodrugs to replace a furanose oxygen of anti-HCV nucleos(t)ides with a selenium atom on the basis that selenium is a chemical isostere of oxygen. These nucleosides are expected to show different physicochemical properties such as better lipophilicity which might enhance the penetration across cell membranes and the conformational constraint induced by a bulky selenium atom in the sugar ring. The 2'-C-methyl-4'-selenopyrimidine and -purine nucleosides 8 and 9 were synthesized from 2-C-methyl-d-ribono-γ-lactone (14) via construction of 2-C-methyl-d-selenosugar 18 through C-4 epimerization and SN2 cyclization with Se2- as key steps. The key 4'-selenosugar was converted to the 2'-C-methyl-4'-selenopyrimidine and -purine nucleosides using Pummerer-type rearrangement and Vorbrüggen glycosylation, respectively. In addition, the ProTide strategy has been applied to synthesize the adenine and uracil phosphoramidate derivatives 10a and 10b to overcome the limitations associated with parent nucleosides such as inefficient conversion to their corresponding 5'-monophosphate form and poor cellular uptake. The regio- and stereochemistry of 4'-selenonucleosides were confirmed by 2D NOESY NMR spectroscopy and X-ray crystallography. None of the final pyrimidine and purine nucleosides and their prodrugs exhibited significant anti-HCV activity up to 100 μM.


Pneumococcal vaccine response in psoriatic arthritis patients during treatment with etanercept.
Therapeutics used to treat inflammatory diseases, including psoriatic arthritis (PsA), may potentially interfere with normal immune system function. Immune system function can be assessed by evaluating response to vaccination. We assessed the ability of patients with PsA treated with etanercept to produce antibodies in response to pneumococcal antigen challenge. Patients with PsA (n = 205) were stratified by methotrexate (MTX) use and randomly assigned to receive either placebo or etanercept 25 mg twice weekly by subcutaneous injection. After 4 weeks of treatment with study drug, a 23-valent pneumococcal vaccination was administered. Antibody levels to 5 antigens (9V, 14, 18C, 19F, and 23F) were measured by ELISA before and 4 weeks after vaccination in 184 patients. The proportion (%) of patients with 2- and 4-fold increases in antibody titers was analyzed. Patients treated with etanercept or placebo had similar responses to the vaccine. A 2-fold increase in titer to at least 2 antigens was achieved by 67% of patients, and a 4-fold increase to at least 2 antigens was achieved by 47% of patients. Approximately 20% of patients in each group failed to show a 2-fold response to any antigens. Logistic regression analysis showed MTX use and age were predictors of a poor response. Patients with PsA treated with etanercept were able to produce antibodies in response to pneumococcal vaccination. Patients receiving MTX had lower mean antibody levels in response to the vaccine. There was no increased risk of poor response with etanercept treatment given alone or with MTX.


The validity and reliability of the Arabic version of the Japanese Orthopedic Association Back Pain Evaluation Questionnaire: Can we implement it in Saudi Arabia?
The Japanese Orthopedic Association designed their back pain evaluation questionnaire (JOABPEQ) which was proven to be feasible, reliable and valid. The questionnaire's validity and reliability were confirmed when it was implemented in patients with different cultural background. Since the JOABPEQ still has not been utilized in any of the Arabic countries, we present the Arabic version with its validity and reliability. After ensuring an optimal forward-backward translation of the JOABPEQ, the Arabic version was distributed among 151 patients visiting our clinics from December 2015 to February 2016. The reliability was tested using Cronbach's alpha. The convergent validity was assessed through aggregating and correlating the questionnaire's items with their previously designed and validated subscales described in the JOABPEQ validity study by Fukui et al. (four items for social function, seven for mental health, six for lumbar function, five for walking ability and four for low back pain); Spearman's correlation matrix was used. The correlation coefficient had to be greater than 0.40 for each item with its corresponding subscale to be satisfactory. Scores underwent descriptive analysis, and Mann-Whitney u test was performed to compare between categorical subgroups. 151 spine patients completed the questionnaire. The mean age (range) was 34.9 (10-72) years. The male respondents were 61 (40.4%) and the females were 90 (59.6%). The result of Cronbach's alpha for internal consistency (reliability) was 0.87 for the 25 items. The validity was confirmed since the correlation coefficient was greater than 0.4 for each item with only its relevant subscale. the Arabic version of the JOABPEQ is valid, reliable and feasible in assessing patients with spine disorders. We believe the JOABPEQ with its different versions are suitable questionnaires to be used across nations and can serve as a unified tool in conducting research and exchanging information in the future.


Behavioural-variant frontotemporal dementia: diagnosis, clinical staging, and management.
Patients with behavioural-variant frontotemporal dementia (bvFTD) present with insidious changes in personality and interpersonal conduct that indicate progressive disintegration of the neural circuits involved in social cognition, emotion regulation, motivation, and decision making. The underlying pathological changes are heterogeneous and are characterised by various intraneuronal inclusions. Biomarkers to detect these histopathological changes in life are becoming increasingly important with the development of disease-modifying drugs. Gene mutations have been found that collectively account for around 10-20% of cases. Recently, criteria proposed for bvFTD define three levels of diagnostic certainty: possible, probable, and definite. Detailed history taking from family members to elicit behavioural features underpins the diagnostic process, with support from neuropsychological testing designed to detect impairment in decision making, emotion processing, and social cognition. Brain imaging is important for increasing the level of diagnostic certainty. A recently developed staging instrument shows much promise for monitoring patients and evaluating therapies, which at present are aimed at symptom amelioration. Carer education and support remain of paramount importance.