Inequalities in access to HIV and syphilis tests in prenatal care in Brazil.
This study aims to evaluate the social determinants of access to HIV and VDRL tests during pregnancy in Brazil. The dependent variables were based on prenatal care access: prenatal care appointments, no HIV and syphilis tests. The independent variables at the first level were formal education level, age, race, work and participation in the Family Income program conditional cash transfer program. The city-level variables were the human development index (HDI), Gini index, and indicators related to health services. An exploratory analysis was performed assessing the effect of each level through prevalence ratios (PR) calculation. A multilevel mixed-effect Poisson regression model was constructed for all outcomes to verify the effect of individual level and with both the individual and contextual levels. Regarding prenatal appointments, the main implicated factors were related to individual socioeconomic position (education level and participation in the Family Income Program conditional cash transfer program), however only HDI maintained significance for the city-level context. The city-level variance dropped from 0.049 to 0.042, indicating an important between-city effect. Regarding the outcomes performing tests in prenatal care, the worst conditions such as contextual (HDI > 0.694, p < 0.001; Gini index ≥ 0.521, p < 0.001) and individual (> 8 years of schooling, p < 0.001) showed a risk effect in the final model. Variables related to health services did not show significant effects. They were associated with individual socioeconomic position and a city-level contextual effect. These findings indicate the importance of strengthening HIV and syphilis infection control programs during pregnancy.


The soft embodiment of culture: camera angles and motion through time and space.
Cultural assumptions about one's relation to others and one's place in the world can be literally embodied in the way one cognitively maps out one's position and motion in time and space. In three experiments, we examined the psychological perspective that Asian American and Euro-American participants embodied as they both comprehended and produced narratives and mapped out metaphors of time and space. In social situations, Euro-American participants were more likely to embody their own perspective and a sense of their own motion (rather than those of a friend), whereas Asian American participants were more likely to embody a friend's perspective and sense of motion (rather than their own). We discuss how these psychological perspectives represent the soft embodiment of culture by implicitly instantiating cultural injunctions (a) to think about how you look to others and to harmonize with them or (b) to know yourself, trust yourself, and act with confidence.


Minimally Invasive Delivery of 3D Shape Recoverable Constructs with Ordered Structures for Tissue Repair.
Minimally invasive procedures are becoming increasingly more common in surgery. However, the biomaterials capable of delivering biomimetic, three-dimensional (3D) functional tissues in a minimally invasive manner and exhibiting ordered structures after delivery are lacking. Herein, we reported the fabrication of gelatin methacryloyl (GelMA)-coated, 3D expanded nanofiber scaffolds, and their potential applications in minimally invasive delivery of 3D functional tissue constructs with ordered structures and clinically appropriate sizes (4 cm × 2 cm × 1.5 mm). GelMA-coated, expanded 3D nanofiber scaffolds produced by combining electrospinning, gas-foaming expansion, hydrogel coating, and cross-linking are extremely shape recoverable after release of compressive strain, displaying a superelastic property. Such scaffolds can be seeded with various types of cells, including dermal fibroblasts, bone marrow-derived mesenchymal stem cells, and human neural stem/precursor cells to form 3D complex tissue constructs. Importantly, the developed 3D tissue constructs can be compressed and loaded into a 4 mm diameter glass tube for minimally invasive delivery without compromising the cell viability. Taken together, the method developed in this study could hold great promise for transplantation of biomimetic, 3D functional tissue constructs with well-organized structures for tissue repair and regeneration using minimally invasive procedures like laparoscopy and thoracoscopy.


A heterodimer of α and β hemoglobin chains functions as an innate anticancer agent.
Metastatic (as well as tumor) microenvironments contain both cancer-promoting and cancer-restraining factors. The balance between these opposing forces determines the fate of cancer cells that disseminate to secondary organ sites. In search for microenvironmental drivers or inhibitors of metastasis, we identified, in a previous study, the beta subunit of hemoglobin (HBB) as a lung-derived antimetastatic factor. In the present study, exploring mechanisms regulating melanoma brain metastasis, we discovered that brain-derived factors restrain proliferation and induce apoptosis and necrosis of brain-metastasizing melanoma cells. Employing various purification procedures, we identified a heterodimer composed of hemoglobin alpha and beta chains that perform these antimetastatic functions. Neither the alpha nor the beta subunit alone was inhibitory. An alpha/beta chain dimer chemically purified from human hemoglobin inhibited the cell viability of primary melanomas, melanoma brain metastasis (MBM), and breast cancer cell lines. The dimer-induced DNA damage, cell cycle arrest at the SubG1 phase, apoptosis, and significant necrosis in four MBM cell lines. Proteomic analysis of dimer-treated MBM cells revealed that the dimer downregulates the expression of BRD4, GAB2, and IRS2 proteins, playing crucial roles in cancer cell sustainability and progression. Thus, we hypothesize that the hemoglobin dimer functions as a resistance factor against brain-metastasizing cancer cells.


Primary pulmonary lymphoma in a patient with advanced AIDS.
Non-Hodgkin's lymphoma (NHL) is an AIDS defining lesion and risk of NHL most likely correlates with the degree of immunosuppression from HIV. Risk of NHL is highest among patients with CD4 count <50 cells/mL. Primary pulmonary lymphoma (PPL) is an infrequent cause of AIDS-related lymphoma. The authors report a patient with advanced AIDS presenting with recurrent fever and pulmonary nodule seen on the CT scan. The patient remained febrile despite being on broad spectrum antibiotics with no clear source of infection. The patient underwent a bronchoscopy with biopsy of the pulmonary lesion which was most consistent with diffuse large B-cell lymphoma. The patient was started on dose-adjusted etoposide, vincristine, doxorubicin, cyclophosphamide and prednisone (EPOCH) and was noted to be afebrile and a repeat CT scan few weeks later showed resolution of her pulmonary nodule. This case highlights the importance of considering NHL in patients with advanced AIDS presenting with pulmonary nodule and fever.


Issues in optimising and standardising the accuracy and utility of the colposcopic examination in the HPV era.
For this tribute to Mario Sideri, we reviewed some of the current issues in colposcopy, many of which we were researching with him. The review concentrates on the impact of HPV testing on cervical screening, specifically on the practice of colposcopy as the major diagnostic procedure in cervical screening programmes. Topics include the changing population of women referred to colposcopy, evolving views of the colposcopic impression, differing approaches to directed and random biopsy, issues in teaching colposcopy using static images, and the development of colposcopy aids, and simplified visual assessment techniques.


Studies on levamisole--induced agranulocytosis.
Widespread clinical trials of leavo-tetramisole (levamisole) as an immunopotentiating agent in rheumatoid arthritis, metastatic carcinoma, and immunodeficiency states have been complicated by agranulocytosis (AGC) in 2.5%-13% of patients. Other than a relationship with prolonged high dosage, very little is known regarding the pathogenesis of levamisole-induced AGC. Whereas leukoagglutination was negative, fluorochromatic microgranulocytotoxicity (GCY) tests were positive with serum from 10 of 10 acutely neutropenic patients. The antibody was IgM, reacted with 100% of unrelated granulocytes, but not with T or B lymphocytes. Some sera also reacted with monocytes and the myeloid cell line, K-562. Tests for antigen-antibody complexes or cold autoantibodies were negative. Although clinical evidence strongly suggests a haptene (drug) mechanism, in vitro mixing experiments were also negative. An alternative choice parallels the model of aldomet-induced Coombs'-positive hemolytic anemia. Finally, GCY first became positive 2-3 mo prior to the onset of AGC on two patients, suggesting the possibility of identifying those at risk well before the onset of neutropenia.


An AML1/ETO fusion transcript is consistently detected by RNA-based polymerase chain reaction in acute myelogenous leukemia containing the (8;21)(q22;q22) translocation.
The 8;21 translocation is one of the most common chromosomal translocations in acute myelogenous leukemia (AML), accounting for 40% of pediatric AML with French-American-British (FAB)-M2 morphology. The chromosomal breakpoints have recently been identified at the molecular level and shown to involve the AML1 gene on chromosome 21 and the ETO gene on chromosome 8. Translocation results in the consistent fusion of these genes on the der(8) chromosome, resulting in the production of a novel chimeric gene and message. Using oligonucleotide primers derived from the AML1 and ETO cDNAs, we were able to amplify a specific fusion transcript from 26 of 26 patients with t(8;21) by a reverse transcriptase polymerase chain reaction (PCR) approach. DNA fragments of identical size were generated from each case including two with complex translocations. Studies on the sensitivity and specificity of this approach show that PCR analysis can be used as a rapid, accurate, and sensitive means for detecting this chromosomal abnormality, and for following the patients' response to therapy.


SU-E-I-22: Metal Artifact Correction Using KV and Selective MV Imaging.
To improve the image quality of radiotherapy planning CTs for patients with metal implants or fillings by completing the missing kV projection data with selectively acquired MV data that does not suffer from photon starvation. Using both imaging systems that are available on current radiotherapy devices, streaking artifacts are avoided and the soft tissue contrast is restored, even in areas where the kV photons do not contribute any information. This enables a better delineation of structures of interest in planning CT images for patients with metal objects. An algorithm for combining kV and MV projection data from the two on-board imagers of a radiotherapy device is presented in this work. It only requires selective MV imaging with the high energy X-rays being collimated onto the metal implants, ensuring that the patient dose does not increase significantly. The algorithm can cope with non-identical geometries of the two imagers and is based on stitching together kV and MV sinograms by estimating a ratio between them. A numerical head phantom with two dental fillings and two soft tissue patterns was used to quantitatively evaluate the proposed hybrid reconstruction algorithm. A structural similarity index (SSIM) with respect to the ground truth data was computed for two ROIs. Realistic, polychromatic spectra were used for both imagers with 120 keV(p) and 6 MeV(p). The patient dose was limited to about 6 cGy for both acquisitions combined. The reconstruction results yield visually as well as objectively better results (SSIM=74.8%) than a simple sinogram interpolation of the kV data (SSIM=69.7%) or a reconstruction from the original data (SSIM=17.9%). We have successfully implemented a new reconstruction method for hybrid kV-MV cone beam CT reconstruction that enables a better planning of radiotherapy treatments for patients with metal implants without compromising their safety. This work was funded by NIH grant 1R01CA138426-01A1.


Counting every stillbirth and neonatal death through mortality audit to improve quality of care for every pregnant woman and her baby.
While there is widespread acknowledgment of the need for improved quality and quantity of information on births and deaths, there has been less movement towards systematically capturing and reviewing the causes and avoidable factors linked to deaths, in order to affect change. This is particularly true for stillbirths and neonatal deaths which can fall between different health care providers and departments. Maternal and perinatal mortality audit applies to two of the five objectives in the Every Newborn Action Plan but data on successful approaches to overcome bottlenecks to scaling up audit are lacking. We reviewed the current evidence for facility-based perinatal mortality audit with a focus on low- and middle-income countries and assessed the status of mortality audit policy and implementation. Based on challenges identified in the literature, key challenges to completing the audit cycle and affecting change were identified across the WHO health system building blocks, along with solutions, in order to inform the process of scaling up this strategy with attention to quality. Maternal death surveillance and review is moving rapidly with many countries enacting and implementing policies and with accountability beyond the single facility conducting the audits. While 51 priority countries report having a policy on maternal death notification in 2014, only 17 countries have a policy for reporting and reviewing stillbirths and neonatal deaths. The existing evidence demonstrates the potential for audit to improve birth outcomes, only if the audit cycle is completed. The primary challenges within the health system building blocks are in the area of leadership and health information. Examples of successful implementation exist from high income countries and select low- and middle-income countries provide valuable learning, especially on the need for leadership for effective audit systems and on the development and the use of clear guidelines and protocols in order to ensure that the audit cycle is completed. Health workers have the power to change health care routines in daily practice, but this must be accompanied by concrete inputs at every level of the health system. The system requires data systems including consistent cause of death classification and use of best practice guidelines to monitor performance, as well as leaders to champion the process, especially to ensure a no-blame environment, and to access change agents at other levels to address larger, systemic challenges.


Genome-wide analysis of WRKY gene family in Cucumis sativus.
WRKY proteins are a large family of transcriptional regulators in higher plant. They are involved in many biological processes, such as plant development, metabolism, and responses to biotic and abiotic stresses. Prior to the present study, only one full-length cucumber WRKY protein had been reported. The recent publication of the draft genome sequence of cucumber allowed us to conduct a genome-wide search for cucumber WRKY proteins, and to compare these positively identified proteins with their homologs in model plants, such as Arabidopsis. We identified a total of 55 WRKY genes in the cucumber genome. According to structural features of their encoded proteins, the cucumber WRKY (CsWRKY) genes were classified into three groups (group 1-3). Analysis of expression profiles of CsWRKY genes indicated that 48 WRKY genes display differential expression either in their transcript abundance or in their expression patterns under normal growth conditions, and 23 WRKY genes were differentially expressed in response to at least one abiotic stresses (cold, drought or salinity). The expression profile of stress-inducible CsWRKY genes were correlated with those of their putative Arabidopsis WRKY (AtWRKY) orthologs, except for the group 3 WRKY genes. Interestingly, duplicated group 3 AtWRKY genes appear to have been under positive selection pressure during evolution. In contrast, there was no evidence of recent gene duplication or positive selection pressure among CsWRKY group 3 genes, which may have led to the expressional divergence of group 3 orthologs. Fifty-five WRKY genes were identified in cucumber and the structure of their encoded proteins, their expression, and their evolution were examined. Considering that there has been extensive expansion of group 3 WRKY genes in angiosperms, the occurrence of different evolutionary events could explain the functional divergence of these genes.


Standardized method for evaluation of hand disinfection by surgical scrub formulations.
A standardized protocol for the evaluation of hand disinfection by surgical scrub formulations was applied to volunteers in a multicenter trial. Povidone iodine (PVI), chlorhexidine (CHX), and a nonmedicated soap (NMS) were tested. The scrubbing procedure involved three daily hand washings for five consecutive days; surviving bacteria were counted daily after being collected in a suitable neutralizing solution. Immediate efficacy (IE), cumulative efficacy (CE), and remanent effect (RE) were calculated by reference to the control hand. Statistical analyses of IE, CE, and RE showed significant differences among the three scrub formulations. IEs of PVI and CHX were equivalent and different from IE of NMS; CE and RE of CHX were higher than those of PVI and NMS. On the basis of the statistical analysis, the population size required for further studies aimed at detecting significant differences between surgical scrub formulations could be estimated.


[Examination and treatment for dilutional thrombocytopenia].
Massive transfusion is defined as transfusion of more than total blood volume within 24 hours. There are several adverse effects associated with massive transfusion, and dilutional thrombocytopenia is known as one of the major adverse effects. Dilutional thrombocytopenia is caused by platelet loss out of the body and platelet dilution with replaced red cells and crystalloids. Volume of blood loss or replaced volume is a good indicator of dilutional thrombocytopenia, and previous reports suggest that severe thrombocytopenia doesn't occur before replaced volume surpasses over one hundred and fifty percent of total blood volume. Recently, an automated blood cell counter has spread and platelet count is available in a short time, even at night. To treat the patient with dilutional thrombocytopenia, platelet count is very helpful to decide when to start platelet transfusion. When platelet count decreases as low as 50,000/mm3, platelet transfusion should be considered. Nowadays, dilutional thrombocytopenia is less frequent complications of massive transfusion than before, because platelet transfusion tends to be performed before platelet count fall to the critical point. Thus, exceeded platelet transfusion might become another problem after massive transfusion.


An Approach Towards Reducing Road Traffic Injuries and Improving Public Health Through Big Data Telematics: A Randomised Controlled Trial Protocol.
Deaths due to road traffic accidents (RTAs) are a major public health concern around the world. Developing countries are over-represented in these statistics. Punitive measures are traditionally employed to lower RTA related behavioural risk factors. These are, however, resource intensive and require infrastructure development. This is a randomised controlled study to investigate the effect of non-punitive behavioural intervention through peer-comparison feedback based on driver behaviour data gathered by an in-vehicle telematics device. A randomised controlled trial using repeated measures design conducted in Iran on the drivers of 112 public transport taxis in Tehran province and 1309 inter-city busses operating nationwide. Driving data is captured by an in-vehicle telematics device and sent to a centrally located data centre using a mobile network. The telematics device is installed in all vehicles. Participants are males aged above 20 who have had the device operating in their vehicles for at least 3 months prior to the start of the trial. The study had three stages: 1- Driver performance was monitored for a 4-week period after which they were randomised into intervention and control groups. 2- Their performance was monitored for a 9-week period. At the end of each week, drivers in the intervention group received a scorecard and a note informing them of their weekly behaviour and ranking within their peer group. Drivers in the control group received no feedback via short messaging service (SMS). 3- Drivers did not receive further feedback and their behaviour was monitored for another 4 weeks. Primary outcome was changes in weekly driving score in intervention and control groups during stage 2 of intervention. Taxis and busses were analysed separately using generalised estimating equation analysis. This project was funded by the National Institute for Medical Research Development (Grant No.940576) and approved by its ethics committee (Code: IR.NIMAD.REC.1394.016). This trial was registered at www.irct.ir as IRCT20180708040391N1.


Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia.
The study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials. Quantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers. The qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range (1.5 x 103 to 1.5 x 108 copies/mL) and correlated well with those from a commercial assay (r = 0.91, (p < 0.001). Viral load levels dropped dramatically but temporarily during and after a short course of lamivudine therapy. HBV DNA was a more reliable indicator of the presence of virus than HBe antigen and was detected in 77.0% (161/209) of HBeAg negative and in all HBeAg positive carriers. This method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.


Peripheral blood neutrophilia as a biomarker of ozone-induced pulmonary inflammation.
Ozone concentrations are predicted to increase over the next 50 years due to global warming and the increased release of precursor chemicals. It is therefore urgent that good, reliable biomarkers are available to quantify the toxicity of this pollutant gas at the population level. Such a biomarker would need to be easily performed, reproducible, economically viable, and reflective of ongoing pathological processes occurring within the lung. We examined whether blood neutrophilia occurred following a controlled ozone challenge and addressed whether this could serve as a biomarker for ozone-induced airway inflammation. Three separate groups of healthy subjects were exposed to ozone (0.2 ppm, 2h) and filtered air (FA) on two separate occasions. Peripheral blood samples were collected and bronchoscopy with biopsy sampling and lavages was performed at 1.5h post exposures in group 1 (n=13), at 6h in group 2 (n=15) and at 18h in group 3 (n=15). Total and differential cell counts were assessed in blood, bronchial tissue and airway lavages. In peripheral blood, we observed fewer neutrophils 1.5h after ozone compared with the parallel air exposure (-1.1±1.0x10(9) cells/L, p<0.01), at 6h neutrophil numbers were increased compared to FA (+1.2±1.3x10(9) cells/L, p<0.01), and at 18h this response had fully attenuated. Ozone induced a peak in neutrophil numbers at 6h post exposure in all compartments examined, with a positive correlation between the response in blood and bronchial biopsies. These data demonstrate a systemic neutrophilia in healthy subjects following an acute ozone exposure, which mirrors the inflammatory response in the lung mucosa and lumen. This relationship suggests that blood neutrophilia could be used as a relatively simple functional biomarker for the effect of ozone on the lung.


NMR and small-angle scattering-based structural analysis of protein complexes in solution.
Structural analysis of multi-domain protein complexes is a key challenge in current biology and a prerequisite for understanding the molecular basis of essential cellular processes. The use of solution techniques is important for characterizing the quaternary arrangements and dynamics of domains and subunits of these complexes. In this respect solution NMR is the only technique that allows atomic- or residue-resolution structure determination and investigation of dynamic properties of multi-domain proteins and their complexes. As experimental NMR data for large protein complexes are sparse, it is advantageous to combine these data with additional information from other solution techniques. Here, the utility and computational approaches of combining solution state NMR with small-angle X-ray and Neutron scattering (SAXS/SANS) experiments for structural analysis of large protein complexes is reviewed. Recent progress in experimental and computational approaches of combining NMR and SAS are discussed and illustrated with recent examples from the literature. The complementary aspects of combining NMR and SAS data for studying multi-domain proteins, i.e. where weakly interacting domains are connected by flexible linkers, are illustrated with the structural analysis of the tandem RNA recognition motif (RRM) domains (RRM1-RRM2) of the human splicing factor U2AF65 bound to a nine-uridine (U9) RNA oligonucleotide.


Chronic treatment with taurine ameliorates diabetes-induced dysfunction of nitric oxide-mediated neurogenic and endothelium-dependent corpus cavernosum relaxation in rats.
This study was aimed to examine the effect of chronic taurine treatment on corpus cavernosum dysfunction in diabetic rats and to investigate possible underlying mechanisms. Thirty male rats were randomized to three groups of 10 each, including control, diabetic, and taurine-treated diabetic. Diabetes was induced in rats by streptozotocin (STZ, single intraperitoneal dose of 50 mg/kg body weight). Taurine was administered orally for 12 weeks (1% w/v in drinking water) from the day on which STZ was injected. At the end of the 12th week, strips of corpus cavernosum were suspended in an organ bath system for functional studies. Nitric oxide (NO)-mediated endothelium-dependent and neurogenic corpus cavernosum relaxation were evaluated by acetylcholine (ACh, 0.1-100 μm) and electrical field stimulation (EFS, 30 V, 5 ms, 2-32 Hz), respectively. The expressions of endothelial nitric oxide synthase (eNOS), phosphorylated eNOS (p-eNOS) (Ser-1177), neuronal nitric oxide synthase (nNOS), NADPH oxidase subunit gp91(phox) , Rho A, and Rho kinase in corpus cavernosum were semi-quantitatively assessed by immunohistochemistry. Induction of diabetes resulted in significant inhibition of NO-mediated endothelium-dependent and neurogenic corpus cavernosum relaxation. Furthermore, eNOS, p-eNOS, and nNOS expressions decreased significantly in diabetic rats compared to controls, while gp91(phox) , RhoA and Rho kinase expressions increased significantly. The diminished relaxation response to ACh and EFS as well as diabetes-related changes in expressions of these proteins in corpus cavernosum of diabetic rats was significantly improved by taurine. Taurine treatment improves NO-mediated relaxations of corpus cavernosum in diabetic rats probably by inhibiting NADPH oxidase/Rho kinase pathways.


DNAH5 is associated with total lung capacity in chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease (COPD) is characterized by expiratory flow limitation, causing air trapping and lung hyperinflation. Hyperinflation leads to reduced exercise tolerance and poor quality of life in COPD patients. Total lung capacity (TLC) is an indicator of hyperinflation particularly in subjects with moderate-to-severe airflow obstruction. The aim of our study was to identify genetic variants associated with TLC in COPD. We performed genome-wide association studies (GWASs) in white subjects from three cohorts: the COPDGene Study; the Evaluation of COPD Longitudinally to Identify Predictive Surrogate Endpoints (ECLIPSE); and GenKOLS (Bergen, Norway). All subjects were current or ex-smokers with at least moderate airflow obstruction, defined by a ratio of forced expiratory volume in 1 second to forced vital capacity (FEV1/FVC) <0.7 and FEV1 < 80% predicted on post-bronchodilator spirometry. TLC was calculated by using volumetric computed tomography scans at full inspiration (TLCCT). Genotyping in each cohort was completed, with statistical imputation of additional markers. To find genetic variants associated with TLCCT, linear regression models were used, with adjustment for age, sex, pack-years of smoking, height, and principal components for genetic ancestry. Results were summarized using fixed-effect meta-analysis. Analysis of a total of 4,543 COPD subjects identified one genome-wide significant locus on chromosome 5p15.2 (rs114929486, β = 0.42L, P = 4.66 × 10-8). In COPD, TLCCT was associated with a SNP in dynein, axonemal, heavy chain 5 (DNAH5), a gene in which genetic variants can cause primary ciliary dyskinesia. DNAH5 could have an effect on hyperinflation in COPD.


Whole body vibration therapy on a treatment bed as additional means to treat postprostatectomy urinary incontinence.
An innovative form of whole body vibration therapy on a treatment bed (Evocell®) to fight against the disabling and isolating symptom of postoperative incontinence in a prostate cancer patient is presented. A supervised program with outpatient active pelvic floor training and a novel form of synchronous high-intensity whole body vibration therapy using the Evocell® device was performed in a patient with postprostatectomy stress urinary incontinence. The patient had previously failed regular pelvic floor exercise. During the intervention, namely a whole body vibration treatment in a lying position on a treatment bed, the patient performed active and passive pelvic floor exercises under professional guidance. Over a period of 6 weeks after starting treatment, the patient regained continence (usage of 1 safety pad). Furthermore, his ability to work increased (return to work) and his ability to attend social activities improved.


FRET and optical trapping reveal mechanisms of actin activation of the power stroke and phosphate release in myosin V.
Myosins generate force and motion by precisely coordinating their mechanical and chemical cycles, but the nature and timing of this coordination remains controversial. We utilized a FRET approach to examine the kinetics of structural changes in the force-generating lever arm in myosin V. We directly compared the FRET results with single-molecule mechanical events examined by optical trapping. We introduced a mutation (S217A) in the conserved switch I region of the active site to examine how myosin couples structural changes in the actin- and nucleotide-binding regions with force generation. Specifically, S217A enhanced the maximum rate of lever arm priming (recovery stroke) while slowing ATP hydrolysis, demonstrating that it uncouples these two steps. We determined that the mutation dramatically slows both actin-induced rotation of the lever arm (power stroke) and phosphate release (≥10-fold), whereas our simulations suggest that the maximum rate of both steps is unchanged by the mutation. Time-resolved FRET revealed that the structure of the pre- and post-power stroke conformations and mole fractions of these conformations were not altered by the mutation. Optical trapping results demonstrated that S217A does not dramatically alter unitary displacements or slow the working stroke rate constant, consistent with the mutation disrupting an actin-induced conformational change prior to the power stroke. We propose that communication between the actin- and nucleotide-binding regions of myosin assures a proper actin-binding interface and active site have formed before producing a power stroke. Variability in this coupling is likely crucial for mediating motor-based functions such as muscle contraction and intracellular transport.


Meta-analysis of microarray studies reveals a novel hematopoietic progenitor cell signature and demonstrates feasibility of inter-platform data integration.
Microarray-based studies of global gene expression (GE) have resulted in a large amount of data that can be mined for further insights into disease and physiology. Meta-analysis of these data is hampered by technical limitations due to many different platforms, gene annotations and probes used in different studies. We tested the feasibility of conducting a meta-analysis of GE studies to determine a transcriptional signature of hematopoietic progenitor and stem cells. Data from studies that used normal bone marrow-derived hematopoietic progenitors was integrated using both RefSeq and UniGene identifiers. We observed that in spite of variability introduced by experimental conditions and different microarray platforms, our meta-analytical approach can distinguish biologically distinct normal tissues by clustering them based on their cell of origin. When studied in terms of disease states, GE studies of leukemias and myelodysplasia progenitors tend to cluster with normal progenitors and remain distinct from other normal tissues, further validating the discriminatory power of this meta-analysis. Furthermore, analysis of 57 normal hematopoietic stem and progenitor cell GE samples was used to determine a gene expression signature characteristic of these cells. Genes that were most uniformly expressed in progenitors and at the same time differentially expressed when compared to other normal tissues were found to be involved in important biological processes such as cell cycle regulation and hematopoiesis. Validation studies using a different microarray platform demonstrated the enrichment of several genes such as SMARCE, Septin 6 and others not previously implicated in hematopoiesis. Most interestingly, alpha-integrin, the only common stemness gene discovered in a recent comparative murine analysis (Science 302(5644):393) was also enriched in our dataset, demonstrating the usefulness of this analytical approach.


Mortality and morbidity in African clawed frogs (Xenopus laevis) associated with construction noise and vibrations.
In Spring 2008, 175 adult female Xenopus laevis were exposed to construction-related vibrations that caused overt water rippling in the frog tanks. The 3 affected tanks were custom-built static, 300-gal 'pond-style' tanks that sat on the floor of the housing room. The water in the tank developed visible ripples as a result of the vibrations transmitted through the floor during jack-hammering in an adjacent room that was approximately 10 ftaway. All frogs in the tanks displayed buoyancy problems, excessive air gulping, and skin sloughing; ultimately 7 frogs died. In addition, these 7 animals were bloated, and 5 of these 7 had regurgitated and everted their stomach and distal esophagus into the oral cavity, resulting in airway obstruction and death. Gross pathologic findings included regurgitation and eversion of the stomach of the distal portion of the esophagus into the oral cavity, obstruction of the airway, and lung overinflation. No significant histologic lesions were observed. Construction vibrations transmitted through the water appeared to have disrupted the mechanoreceptive function of the lateral line system, resulting in overstimulation of the noxious feeding response, regurgitation, and eversion of the stomach and distal esophagus into the oral cavity and subsequent suffocation due to airway obstruction. After immediate cessation of the jack-hammering and relocation of the remaining frogs, no additional morbidities or mortalities occurred.


Exploring similarities and differences among the self-reported academic integrity of Australian occupational therapy domestic and international students.
Research into the prevalence of dishonest academic behaviours suggests that such behaviours may be extensive among international students in higher education. To compare the academic integrity of domestic and international occupational therapy students and identify possible contributors to students' engagement in dishonest academic behaviours. A quantitative cross-sectional study using a survey containing several standardised scales was used to collect the data. Five Australian universities. 701 occupational therapy students. Students completed a self-report questionnaire comprising demographic questions and six standardised scales: Academic Dishonesty Scale; Academic Dishonesty in the Classroom Setting Scale (ADCS); Academic Dishonesty in the Clinical/Practice Education Setting Scale; Moral Development Scale for Professionals; Academic Dishonesty Tendency Scale; and Perceived Academic Sources of Stress. One-way analysis of variance (ANOVA) was conducted to compare the scores of domestic and international students on academic dishonesty factors, moral development, and perceived sources of academic stress. Significant differences between domestic and international students were observed on age; grade point average; hours per week spent in paid work; moral practice; tendency towards cheating; tendency towards cheating in research assignments; pressures to perform; and self-perception. No significant differences were found on students' self-reported scores on academic dishonesty in academic classroom education contexts, fieldwork practice education settings or perceived stresses of workload and examinations and time restraints. The reported incidences of dishonest academic behaviours in the classroom and fieldwork settings were low for all occupational therapy students, irrespective of origin and were comparable to findings from cohorts of other health professional students. However, concomitant with existing research, international students were found to have a greater tendency to engage in dishonest behaviours particularly on research assignments. This may be reflective the socio-cultural issues and external pressures that international students experience. Further study in this area is recommended.


Radical Approach to Enzymatic β-Thioether Bond Formation.
Ribosomally synthesized and post-translationally modified peptides (RiPPs) are an emerging class of natural products that harbor diverse chemical functionalities, usually introduced via the action of a small number of tailoring enzymes. We have been interested in RiPP biosynthetic gene clusters that encode unusual metalloenzymes, as these may install as yet unknown alterations. Using a new bioinformatic search strategy, we recently identified an array of unexplored RiPP gene clusters that are quorum sensing-regulated and contain one or more uncharacterized radical S-adenosylmethionine (RaS) metalloenzymes. Herein, we investigate the reaction of one of these RaS enzymes and find that it installs an intramolecular β-thioether bond onto its substrate peptide by connecting a Cys-thiol group to the β-carbon of an upstream Asn residue. The enzyme responsible, NxxcB, accepts several amino acids in place of Asn and introduces unnatural β-thioether linkages at unactivated positions. This new transformation adds to the growing list of Nature's peptide macrocyclization strategies and expands the already impressive catalytic repertoire of the RaS enzyme superfamily.


Gene clustering in plant specialized metabolism.
Physically linked clusters of genes that encode the enzymatic information for the synthesis of specialized metabolites are a well-established feature of microbial secondary metabolism. In contrast, the biosynthesis of plant specialized metabolites has until recently been thought to be almost exclusively encoded by genes that are randomly scattered in the genome. However, recent reports highlight the growing number of examples of gene clusters for specialized metabolic pathways in plants. Numerous gene clusters that encode for the biosynthesis of different classes of metabolite have now been discovered in a variety of plant species. Comparison of these characterized clusters now enables us to begin to define their salient features and to exploit plant biosynthetic gene clusters for synthetic biology applications.


Candidate gene polymorphisms and the 9p21 locus in acute coronary syndromes.
It is now widely accepted that the classic environmental risk factors for atherosclerosis only partly explain the incidence of coronary artery disease and the development of acute coronary syndromes. Therefore, genetic factors that vary among human populations seem to be involved in the clinical manifestations of such patients. Substantial data suggest that a significant proportion of genetic polymorphisms involved in endothelial function, inflammation, lipid metabolism, thrombosis and fibrinolysis are often present in patients with acute coronary syndromes. In particular, a common variant on chromosome 9p21 was recently identified to affect the risk of myocardial infarction. Here, we review the progress of candidate gene studies and genome-wide association studies in identifying the genetic bases of complex cardiovascular diseases such as acute coronary syndromes.


Beyond GWAS in COPD: probing the landscape between gene-set associations, genome-wide associations and protein-protein interaction networks.
To use a systems biology approach to integrate genotype and protein-protein interaction (PPI) data to identify disease network modules associated with chronic obstructive pulmonary disease (COPD) and to perform traditional pathway analysis. We utilized a standard gene-set association approach (FORGE) using gene-based association analysis and gene-set definitions from the molecular signatures database (MSigDB). As a discovery step, we analyzed GWAS results from 2 well-characterized COPD cohorts: COPDGene and GenKOLS. We used a third well-characterized COPD case-control cohort for replication: ECLIPSE. Next, we used dmGWAS, a method that integrates GWAS results with PPI, to identify COPD disease modules. No gene-sets reached experiment-wide significance in either discovery population. We identified a consensus network of 10 genes identified in modules by integrating GWAS results with PPI that replicated in COPDGene, GenKOLS, and ECLIPSE. Members of 4 gene-sets were enriched among these 10 genes: (i) lung adenocarcinoma tumor-sequencing genes, (ii) IL-7 pathway genes, (iii) kidney cell response to arsenic, and (iv) CD4 T-cell responses. Further, several genes have also been associated with pathophysiology relevant to COPD including KCNK3, NEDD4L, and RIN3. In particular, KCNK3 has been associated with pulmonary arterial hypertension, a common complication in advanced COPD. We report a set of new genes that may influence the etiology of COPD that would not have been identified using traditional GWAS and pathway analyses alone.


User characteristics and oral contraceptive compliance in Egypt.
Results from the 1988 Egypt Demographic and Health Survey show that many women are not taking oral contraceptives in a manner that ensures full protection by the method. Reports from 1,258 current pill users show a range of incorrect use; 63 percent of women surveyed reported an interruption in their use of the pill in the past month, and of those women, only 40 percent took the correct action when they missed a pill. The majority (89 percent) did not wait the correct number of days between packs. Multivariate analysis revealed that rural women were more likely to take pills out of sequence, compared with their urban counterparts. Failure to take a pill within the previous month was strongly associated with the experience of side effects. The younger women surveyed were more likely to know the correct interval between pill packs than were older women; and women who wanted more children were more likely to know the correct interval than those who did not. The use patterns exhibited by the pill users may be the result of their receiving confusing, incorrect, or incomplete information, and highlight the need to provide women with accurate, updated, and comprehensible information about oral contraceptives.


Neurofibromatosis 2011: a report of the Children's Tumor Foundation annual meeting.
The 2011 annual meeting of the Children's Tumor Foundation, the annual gathering of the neurofibromatosis (NF) research and clinical communities, was attended by 330 participants who discussed integration of new signaling pathways into NF research, the appreciation for NF mutations in sporadic cancers, and an expanding pre-clinical and clinical agenda. NF1, NF2, and schwannomatosis collectively affect approximately 100,000 persons in US, and result from mutations in different genes. Benign tumors of NF1 (neurofibroma and optic pathway glioma) and NF2 (schwannoma, ependymoma, and meningioma) and schwannomatosis (schwannoma) can cause significant morbidity, and there are no proven drug treatments for any form of NF. Each disorder is associated with additional manifestations causing morbidity. The research presentations described in this review covered basic science, preclinical testing, and results from clinical trials, and demonstrate the remarkable strides being taken toward understanding of and progress toward treatments for these disorders based on the close interaction among scientists and clinicians.


Highly stable selenadiazole derivatives induce bladder cancer cell apoptosis and inhibit cell migration and invasion through the activation of ROS-mediated signaling pathways.
Bladder cancer is still a common malignancy of the urinary tract due to the high metastasis rate and unexpected side effects of drug treatments. The acidic environment of the urinary bladder also strongly limits the efficacy of the chemotherapeutic agents during the treatment of bladder cancer. In this study, a series of selenadiazole derivatives (SeDs) have been rationally designed and synthesized and could actively suppress the progression and metastasis of bladder cancer cells. SeDs demonstrated better antiproliferative activity and higher stability under different physiological conditions, especially in an acidic urocystic environment, than mitomycin, a clinically used anti-bladder cancer drug. In particular, compound 1b displayed better selectivity between cancer and normal cells in comparison with other compounds. Studies on the structure-activity relationship revealed that the introduction of strong electron donating substituents, such as the methoxy group, resulted in a dramatic enhancement in the anticancer efficacy. Furthermore, 1b induced anti-migration and anti-invasion activities against bladder cancer cells. Mechanistic investigation revealed that compound 1b was able to enter the cells through endocytosis and then trigger reactive oxygen species (ROS) overproduction, further causing DNA damage-mediated p53 phosphorylation and promoting cancer cell apoptosis by regulating the AKT and MAPKs signaling pathways. Altogether, the study provides a strategy for rational design of selenadiazole derivatives with improved stability to antagonize bladder cancer.


Acceptability and feasibility of mifepristone-misoprostol for menstrual regulation in Bangladesh.
Annually, more than 700,000 women turn to menstrual regulation, or uterine evacuation with vacuum aspiration; many more resort to unsafe abortion. Using pills for the evacuation of the uterus could increase women's access to safe menstrual regulation services and reduce the high levels of abortion- and menstrual regulation- related morbidity in Bangladesh. At 10 facilities in Bangladesh, 651 consenting women who were seeking menstrual regulation services and who were 63 days or less past their last menstrual period received 200 mg of mifepristone followed 24 hours later by 800 mcg of buccal misoprostol, administered either at home or in the clinic. Prospective data were collected to determine women's experience and satisfaction with the procedure, menstrual regulation outcome, and the human and physical resources required for providing the method. Focus group discussions were conducted with a purposively sampled group of service providers at each site to understand their attitudes about the introduction of menstrual regulation with medication. The majority of women (93%) with known menstrual regulation outcomes evacuated the uterus without surgical intervention. Overall, most women (92%) were satisfied with use of pills for their menstrual regulation. Providers faced initial challenges and concerns, particularly related to the additional counseling requirements and lack of control over the final outcome, but became more confident after successful use of the medication regimen. Mifepristone-misoprostol can be safely offered within existing menstrual regulation services in urban and periurban areas in Bangladesh and is highly acceptable to women. Providers' initial concerns diminish with increased experience with the method.


Regulation of axonal caliber, neurofilament content, and nuclear localization in mature sensory neurons by nerve growth factor.
The neuronal perikaryal response to axonal injury (axon reaction) includes reduction in axonal caliber beginning in the proximal portion of the nerve (somatofugal axonal atrophy), development of nuclear eccentricity, and chromatolysis. The means by which these events are triggered is unknown, but it has been argued that loss of a neurotrophic signal from the target of injured neurons plays a role. To date, the identity of this substance(s) remains unknown. In the present study, we have asked whether NGF normally functions to control axonal caliber of sensory neurons in the L4 and L5 dorsal root ganglia (DRG) of the adult rat. Two approaches were used: (1) NGF was continuously delivered to the proximal stump of a transected sciatic nerve to determine whether NGF administration would prevent the production of somatofugal axonal atrophy; and (2) NGF antisera were administered to normal animals to determine whether NGF deprivation would produce somatofugal axonal atrophy. In the first experiment, 9-week-old rats underwent a unilateral sciatic nerve transection at midthigh, and the proximal stump was connected to an osmotic pump containing either NGF or cytochrome C (as control). At 11 weeks of age, dorsal root fibers in lumbar DRG from the control group appeared smaller in caliber and less circular in shape than fibers from age-matched normal animals. Although smaller than those in normal animals, fibers from the NGF-treated nerves were larger than in axotomized controls. Mean axonal area and shape factor (an index in circularity) were measured and found to be decreased significantly (22% and 15%, respectively) from the control group. Fibers from the NGF-treated nerves were significantly (p less than 0.05) larger in axonal caliber and more circular in shape; mean values were only reduced by 11% and 10%, respectively. Quantitation of neurofilament (NF) numbers revealed that the larger calibers in the NGF-treated nerves result from a greater NF content. NGF treatment did not prevent the atrophy of motor fibers in the proximal ventral root. In the second experiment, 2 antisera to mouse NGF were given daily into the footpad for 11 or 12 d; control animals were given normal goat serum. Quantitation of axonal calibers in the L5 DRG demonstrated that mean axonal area and shape factor were significantly (p less than 0.05) reduced by 14% and 17% respectively. The axoplasm of atrophic fibers demonstrated a paucity of NFs.(ABSTRACT TRUNCATED AT 400 WORDS)


Patient-centred dietetic care from the perspectives of older malnourished patients.
Governing organisations for health services currently recommend a patient-centred (PC) approach to practice for all health professions, including dietetics. For the vulnerable older malnourished patient, this approach needs to be prioritised to improve outcomes. The paucity of patient experience data likely limits evidence-based, patient-centred care (PCC) from being implemented effectively. The present study aimed to identify quality indicators of dietetic services from the perspectives of older malnourished patients to inform evidence-based PC dietetic care. Surveys were completed by a sample of 28 females and 28 males (mean age 81 years) who had been seen by a dietitian for malnutrition assessment. In-depth, face-to-face, semi-structured interviews were undertaken with a sub-sample of four females and six males (mean age 81 years). Interviews were transcribed verbatim. Thematic analysis of transcripts and open-ended survey responses was conducted to determine patient-identified quality indicators. Three structure indicators (continuity of care through regular contact and post-discharge dietetic follow-up; interdisciplinary coordination and collaboration; and high-quality hospital food services), five process indicators (addressing a patient's primary medical concern; involving the patient's family; providing clear and simple dietetic information; providing expert dietary knowledge; utilising interpersonal communication skills) and three outcome indicators (improvement in health status; improvement or maintenance of independence; weight gain) were identified. The experiences of older malnourished patients with dietetic services, as described in the present study, reinforce the importance of ensuring high-quality and tailored dietetic care as a key element of PC dietetic services. The quality indicators of dietetic services identified in the present study may facilitate dietitians to provide evidence-based PCC for older malnourished patients.


Excitatory synaptic drive for swimming mediated by amino acid receptors in the lamprey.
In order to investigate the properties and pharmacology of the excitatory synaptic drive received by motoneurons during swimming in the lamprey, propriospinal excitatory interneurons were activated as a population by the regional application of N-methyl-D,L-aspartate (NMA) to either the 6-8 rostral-most or the 6-8 caudal-most segments of lengths of isolated spinal cord. This caused a rhythmic motor output to be generated in these regions. Synaptic potentials that were phase-locked to, and dependent on, the rhythmic motor activity of the segments exposed to the agonist could be recorded in motoneurons lying outside the activated regions. The synaptic drive to motoneurons located rostral or caudal to the activated regions was studied. Motoneurons received both descending and ascending synaptic input, which consisted of alternating excitatory and inhibitory phases. The inhibition could be reversed by chloride injection and blocked by strychnine, leaving an oscillating excitatory phase. The descending excitatory drive could extend 1-9 segments from the active region, while the ascending excitatory drive was recorded only in motoneurons that were 1-3 segments rostral to the active region. Both types of drive occurred in phase with the ipsilateral ventral root discharge: The peak depolarization of the descending drive occurred at the same point in the swimming cycle as that of the depolarizing phase seen during fictive swimming, while that of the ascending drive occurred significantly later. Both ascending and descending drives were partially reduced in amplitude by 2-amino-5-phosphonovaleric acid or Mg2+. The blocking action of Mg2+ was, in both cases, voltage dependent. Cis-2,3-piperidine dicarboxylic acid or kynurenic acid caused a much greater reduction in the amplitude of the oscillations. These results suggest that a major part of the excitatory drive for swimming in lamprey motoneurons is generated by populations of propriospinal interneurons with relatively long descending and/or short ascending axons, which fire rhythmically during swimming and release an amino acid transmitter that excites motoneurons through N-methyl-D-aspartate (NMDA) and non-NMDA receptors. This information will allow these important neurons to be identified in future experiments.


The effects of direct brain stimulation in humans depend on frequency, amplitude, and white-matter proximity.
Researchers have used direct electrical brain stimulation to treat a range of neurological and psychiatric disorders. However, for brain stimulation to be maximally effective, clinicians and researchers should optimize stimulation parameters according to desired outcomes. The goal of our large-scale study was to comprehensively evaluate the effects of stimulation at different parameters and locations on neuronal activity across the human brain. To examine how different kinds of stimulation affect human brain activity, we compared the changes in neuronal activity that resulted from stimulation at a range of frequencies, amplitudes, and locations with direct human brain recordings. We recorded human brain activity directly with electrodes that were implanted in widespread regions across 106 neurosurgical epilepsy patients while systematically stimulating across a range of parameters and locations. Overall, stimulation most often had an inhibitory effect on neuronal activity, consistent with earlier work. When stimulation excited neuronal activity, it most often occurred from high-frequency stimulation. These effects were modulated by the location of the stimulating electrode, with stimulation sites near white matter more likely to cause excitation and sites near gray matter more likely to inhibit neuronal activity. By characterizing how different stimulation parameters produced specific neuronal activity patterns on a large scale, our results provide an electrophysiological framework that clinicians and researchers may consider when designing stimulation protocols to cause precisely targeted changes in human brain activity.


An introduction to the clinical toxicology of volatile substances.
Acute poisoning with organic solvents and other volatile compounds now usually follows deliberate inhalation (volatile substance abuse) or ingestion of these compounds. Solvents from adhesives, typewriter correction and dry cleaning fluids, cigarette lighter refills (butane) and aerosol propellants are commonly abused. The major risk is that of sudden death. Arrhythmias leading to cardiac arrest are thought to cause most deaths, but anoxia, respiratory depression and vagal stimulation leading to cardiac arrest may also contribute, as may indirect causes such as aspiration of vomit or trauma. In the United Kingdom (UK), 3.5 to 10% of young people have at least experimented with volatile substance abuse and mortality is more than 100 per annum. The products abused are cheap and readily available despite legislation designed to limit supply. Volatile substance abuse is not illegal and only a minority of abusers are known to progress to heavy alcohol or illicit drug use. Prevention of abuse by education, not only of children but also of parents, teachers, retailers and health care workers, is important in limiting the problem. However, volatile substance abuse-related deaths are still increasing in the UK despite many measures aimed at prevention. Clinically, volatile substance abuse is characterised by a rapid onset of intoxication and rapid recovery. Euphoria and disinhibition may be followed by hallucinations, tinnitus, ataxia, confusion, nausea and vomiting. It is important not to further alarm the patient if signs of serious toxicity are present, since a cardiac arrest may be precipitated. Further exposure should be prevented and the patient resuscitated and given supplemental oxygen if necessary. Cardiac arrhythmias should be treated conventionally and respiratory failure managed supportively. Long term exposure to n-hexane is associated with the development of peripheral neuropathy, while prolonged abuse (notably of toluene or chlorinated solvents) can cause permanent damage to the central nervous system, heart, liver, kidney and lungs. Knowledge of the routes of absorption, distribution and excretion of volatile compounds, and of the rates governing these processes, is important in understanding the rate of onset, intensity and duration of intoxication, and rate of recovery after volatile substance abuse. In addition, such knowledge is helpful when the clinician is attempting to interpret the results of toxicological analyses performed on samples (blood, other tissues, urine) from such patients. Many volatile substances are partly metabolised, the metabolites being eliminated in exhaled air or in urine. Although metabolism normally results in detoxification, enhanced toxicity may also result as with carbon tetrachloride, chloroform, dichloromethane, n-hexane, trichloroethylene and possibly halothane.(ABSTRACT TRUNCATED AT 400 WORDS)


In vivo reflectance confocal microscopy of shave biopsy wounds: feasibility of intraoperative mapping of cancer margins.
Reflectance confocal microscopy (RCM) images skin at cellular resolution and has shown utility for the diagnosis of nonmelanoma skin cancer in vivo. Topical application of aluminium chloride (AlCl(3)) enhances contrast in RCM images by brightening nuclei. To investigate feasibility of RCM imaging of shave biopsy wounds using AlCl(3) as a contrast agent. AlCl(3) staining was optimized, in terms of concentration vs. immersion time, on excised tissue ex vivo. RCM imaging protocol was tested in patients undergoing shave biopsies. The RCM images were retrospectively analysed and compared with the corresponding histopathology. For 35% AlCl(3) , routinely used for haemostasis in clinic, minimum immersion time was determined to be 1 min. We identified three consistent patterns of margins on RCM mosaic images by varying depth: epidermal margins, peripheral dermal margins, and deep dermal margins. Tumour islands of basal cell carcinoma were identified at peripheral or deep dermal margins, correlating on histopathology with aggregates of neoplastic basaloid cells. Atypical cobblestone or honeycomb patterns were identified at the epidermal margins in squamous cell carcinomas, correlating with a proliferation of atypical keratinocytes extending to biopsy margins. RCM imaging of shave biopsy wounds is feasible and demonstrates the future possibility of intraoperative mapping in surgical wounds.


Treatment outcomes of patients with non-bacteremic pneumonia caused by extensively drug-resistant Acinetobacter calcoaceticus-Acinetobacter baumannii complex isolates: Is there any benefit of adding tigecycline to aerosolized colistimethate sodium?
Few therapeutic options exist for various infections caused by extensively drug-resistant Acinetobacter calcoaceticus-Acinetobacter baumannii (XDR-Acb) complex isolates, including pneumonia. This study investigated the clinical efficacy between aerosolized colistimethate sodium (AS-CMS, 2 million units thrice a day) treatment alone or in combination with standard-dose tigecycline (TGC) in patients with non-bacteremic pneumonia due to XDR-Acb, and explored the factors influencing patients' 30-day mortality.A 1:1 case (n = 106; receiving TGC plus AS-CMS) control (receiving AS-CMS alone with matching scores) observational study was conducted among adult patients with non-bacteremic XDR-Acb complex pneumonia in a Taiwanese medical center from January 2014 through December 2016. The clinically relevant data were retrospectively recorded. The primary endpoint was 30-day case fatality. Secondary endpoints investigated that if the co-morbidities, XDR-A. baumannii as a pneumonic pathogen, therapy-related factors, or airway colonization with colistin-resistant Acb negatively influenced the 14-day clinical condition of enrolled patients.A higher 30-day mortality rate was noted among the group receiving combination therapy (34.0% vs 22.6%; P = .17). The ≥7-day AS-CMS therapy successfully eradicated > 90% of airway XDR-Acb isolates. Nevertheless, follow-up sputum specimens from 10 (6.4% [10/156]) patients were colonized with colistin-resistant Acb isolates. After the conditional factors were adjusted by multivariate logistic analysis, the only factor independently predicting the 30-day case-fatality was the failure of treating XDR-Acb pneumonia at 14 days (adjusted odds ratio [aOR] = 38.2; 95% confidence interval [CI] = 9.96-142.29; P < .001). Cox proportional regression analysis found that chronic obstructive pulmonary disease (COPD) (adjusted hazard ratio [aHR] = 2.08; 95% CI = 1.05-4.10; P = .035), chronic renal failure (aHR = 3.00; 95% CI = 1.52-5.90; P = .002), non-invasive ventilation use (aHR = 2.68; 95% CI = 1.37-5.25; P = .004), and lack of TGC therapy (aHR = 0.52; 95% CI = 0.27-1.00; P = .049) adversely influenced the 14-day clinical outcomes. Conversely, the emergence of colistin-resistant Acb isolates in the follow-up sputum samples was not statistically significantly associated with curing or improving XDR-Acb pneumonia.In conclusion, aggressive pulmonary hygiene care, the addition of TGC, and corticosteroid dose tapering were beneficial in improving the 14-day patients' outcomes.


Postpartum Sequelae of the Hypertensive Diseases of Pregnancy: A Pilot Study.
Hypertensive disorders are one of the most common medical conditions that may complicate pregnancy. Postpartum blood pressure (BP) pattern is, however, less clear in affected women and decision to discharge them is usually decided arbitrarily. A cohort study conducted at Lagos University Teaching Hospital, Lagos, Nigeria, aimed at determining the proportion of pregnant women with pregnancy-induced hypertension (PIH) and preeclampsia (PE) whose BP remains elevated 6 weeks postdelivery and factors associated with the persistent rise. Fifteen women each with PIH and PE were studied in different phases of pregnancy and followed up until 6 weeks postdelivery. Fifteen normotensive pregnant women served as controls. BP patterns were monitored and fasting lipid levels, serum creatinine, fasting glucose profile (FGP), and FGP/insulin ratio were assayed. Data were analyzed with IBM SPSS version 20. Proportion of women with PIH or PE who had persistent hypertension at 6 weeks postpartum was 3/29 (10.3%), risk ratio of 1.1. No statistically significant association was found between mean arterial BP at 6 weeks postpartum and age, parity, gestational age at delivery, body mass index, and family history of hypertension. Serum creatinine level showed moderate correlation with persistent hypertension at 6 weeks postpartum (r = 0.441,P = 0.006), with sensitivity of 100% and specificity of 81.8% at cutoff value of 1.2 mg/dL in detecting pregnant women with hypertensive disorders who will likely remain hypertensive at 6 weeks postpartum. There is a need for long-term follow-up of women with PIH/PE beyond puerperium.


CAS and PSI increase coronal alignment accuracy and reduce outliers when compared to traditional technique of medial open wedge high tibial osteotomy: a meta-analysis.
Medial open-wedge high tibial osteotomy (MOWHTO) is an accepted option in the treatment of medial compartment osteoarthritis of the knee in young and active patients. Functional results are closely correlated to the correction of the mechanical axis of the lower limb. Although several angular and geometrical methods and values have been proposed in the past, the ideal target is still debated. In addition, it is important to have a deep correlation between the planned correction and the achieved correction after surgery. The aim of the present systematic review was to identify the ideal coronal correction after MOWHTO and the most accurate method to achieve it. A systematic review of the literature was completed on July 3rd 2020 in the Pubmed, Medline, Cochrane Reviews, and Google Scholar databases using the Medical Subject Headings (MeSH) terms: "high tibial osteotomy" AND "accuracy" OR "planning". 28 studies were included; 18 were focused on computer-assisted surgery (CAS) and 10 on patient-specific instrumentation (PSI). There were 598 patients in the CAS group and 501 in the control group; the rate of outliers was 16% and 38.2% respectively (P = 0.04), while there was no significant difference between the two groups (SMD = - 0.10; 95% CI 1.31 to 1.12; P = n.s.) in terms of coronal accuracy. Likewise, there were 318 patients in the PSI group and 40 in the control group; the rate of outliers was 15% and 40% respectively (P = 0.98), while there was no significant difference between the two groups (SMD = 0.01; 95% CI 0.58 to 0.59; P = 0.98). A statistically significant reduced outlier rate and a non-significant increased accuracy emerged with the use of CAS when compared to the traditional surgical technique, whereas the results of PSI were still inconclusive. In addition, it emerged clearly that no consensus still exists on the ideal correction target to be achieved after surgery. III.


Willingness to pay for community-based health insurance in Nigeria: do economic status and place of residence matter?
We examine socio-economic status (SES) and geographic differences in willingness of respondents to pay for community-based health insurance (CBHI). The study took place in Anambra and Enugu states, south-east Nigeria. It involved a rural, an urban and a semi-urban community in each of the two states. A pre-tested interviewer-administered questionnaire was used to collect information from a total of 3070 households selected by simple random sampling. Contingent valuation was used to elicit willingness to pay (WTP) using the bidding game format. Data were examined for correlation between SES and geographic locations with WTP. Log ordinary least squares (OLS) was used to examine the construct validity of elicited WTP. Generally, less than 40% of the respondents were willing to pay for CBHI membership for themselves or other household members. The proportions of people who were willing to pay were much lower in the rural communities, at less than 7%. The average that respondents were willing to pay as a monthly premium for themselves ranged from 250 Naira (US$1.7) in a rural community to 343 Naira (US$2.9) in an urban community. The higher the SES group, the higher the stated WTP amount. Similarly, the urbanites stated higher WTP compared with peri-urban and rural dwellers. Males and people with more education stated higher WTP values than females and those with less education. Log OLS also showed that previously paying out-of-pocket for health care was negatively related to WTP. Previously paying for health care using any health insurance mechanism was positively related to WTP. Economic status and place of residence amongst other factors matter in peoples' WTP for CBHI membership. Consumer awareness has to be created about the benefits of CBHI, especially in rural areas, and the amount to be paid has to be augmented with other means of financing (e.g. government and/or donor subsidies) to ensure success and sustainability of CBHI schemes.


Experimental Evidence that Stochasticity Contributes to Bacterial Composition and Functioning in a Decomposer Community.
Stochasticity emerging from random differences in replication, death, mutation, and dispersal is thought to alter the composition of ecological communities. However, the importance of stochastic effects remains somewhat speculative because stochasticity is not directly measured but is instead inferred from unexplained variations in beta-diversity. Here, we performed a field experiment to more directly disentangle the role of stochastic processes, environmental selection, and dispersal in the composition and functioning of a natural bacterial decomposer community in the field. To increase our ability to detect these effects, we reduced initial biological and environmental heterogeneity using replicate nylon litterbags in the field. We then applied two treatments: ambient/added precipitation and bacterial and fungal dispersal using "open" litterbags (made from 18.0-μm-pore-size mesh) ("open bacterial dispersal") versus bacterial and fungal dispersal using "closed" litterbags (made from 22.0-μm-pore-size mesh) ("closed bacterial dispersal"). After 5 months, we assayed composition and functioning by the use of three subsamples from each litterbag to disentangle stochastic effects from residual variation. Our results indicate that stochasticity via ecological drift can contribute to beta-diversity in bacterial communities. However, residual variation, which had previously been included in stochasticity estimates, accounted for more than four times as much variability. At the same time, stochastic effects on beta-diversity were not attenuated at the functional level, as measured by genetic functional potential and extracellular enzyme activity. Finally, dispersal was found to interact with precipitation availability to influence the degree to which stochasticity contributed to functional variation. Together, our results demonstrate that the ability to quantify stochastic processes is key to understanding microbial diversity and its role in ecosystem functioning.IMPORTANCE Randomness can alter the diversity and composition of ecological communities. Such stochasticity may therefore obscure the relationship between the environment and community composition and hinder our ability to predict the relationship between biodiversity and ecosystem functioning. This study investigated the role of stochastic processes, environmental selection, and dispersal in microbial composition and its functioning on an intact field community. To do this, we used a controlled and replicated experiment that was similar to that used to study population genetics in the laboratory. Our study showed that, while the stochastic effects on taxonomic composition are smaller than expected, the degree to which stochasticity contributes to variability in ecosystem processes may be much higher than previously assumed.


The phenotypes of ATG9, ATG16 and ATG9/16 knock-out mutants imply autophagy-dependent and -independent functions.
Macroautophagy is a highly conserved intracellular bulk degradation system of all eukaryotic cells. It is governed by a large number of autophagy proteins (ATGs) and is crucial for many cellular processes. Here, we describe the phenotypes of Dictyostelium discoideum ATG16(-) and ATG9(-)/16(-) cells and compare them to the previously reported ATG9(-) mutant. ATG16 deficiency caused an increase in the expression of several core autophagy genes, among them atg9 and the two atg8 paralogues. The single and double ATG9 and ATG16 knock-out mutants had complex phenotypes and displayed severe and comparable defects in pinocytosis and phagocytosis. Uptake of Legionella pneumophila was reduced. In addition, ATG9(-) and ATG16(-) cells had dramatic defects in autophagy, development and proteasomal activity which were much more severe in the ATG9(-)/16(-) double mutant. Mutant cells showed an increase in poly-ubiquitinated proteins and contained large ubiquitin-positive protein aggregates which partially co-localized with ATG16-GFP in ATG9(-)/16(-) cells. The more severe autophagic, developmental and proteasomal phenotypes of ATG9(-)/16(-) cells imply that ATG9 and ATG16 probably function in parallel in autophagy and have in addition autophagy-independent functions in further cellular processes.


Are there secular trends in the occurrence and severity of rheumatoid arthritis?
There are considerable research and health service benefits in understanding current trends in a disease as common and as severe as rheumatoid arthritis. Unfortunately in the absence of both a high and constant case fatality rate and a population based morbidity registration system, there is no direct means of assessing changes in incidence in contrast, for example, to the situation with cancer. All the available data are inadequate and subject to considerable bias. It is of note however that all the available data are relatively consistent in pointing to a recent reduction in incidence over the last two decades coupled with a decrease in case severity. The immediate problem remains therefore to set up a true population based recording system which will permit the identification of trends both of incidence and severity. It should not be necessary for this to be established nationwide and a study area within a country could be chosen providing its population was representative and of sufficient size. For the latter a minimum seize of 500,000 persons would be needed.


[Survey of MRSA infection in Ibaraki-ken--Ibaraki Association for infectious diseases].
This study which was undertaken at several major hospitals in Ibaraki-ken revealed that, 1) the proportion of Staphylococcus aureus in all the isolates ranged from 5% to 17% and the prevalence of MRSA in isolates of Staphylococcus aureus varied from 51% to 82% depending upon each individual hospital. 2) the clinical response to FOM and CMZ combination therapy was 88.9% and, to FOM and CZON was 57.1% showing no statistical differences between the two regimens. 3) in vitro analyses assessed by the FIC index of 251 isolates revealed that FOM had an additive or synergistic effect with CMZ in 53.8%, with CZON in 66.9%, with minocycline (MINO) in 43.8%, with cefamandole (CMD) in 61.8%, with cefazolin (CEZ) in 62.9% and with imipenem/cliastatin sodium (IPM/CS) in 68.9% at all isolates. 4) the cumulative curve of susceptibility to single CMZ and the combination of CMZ and FOM revealed that the blood levels of CMZ at 3 hours after intravenous administration covered 57% of isolates when used alone and 82% of isolates when in combination with FOM. The blood levels of CZON at 3 hours covered 5% of isolates when used alone and 41% when in combination with FOM. The extent of the therapeutic effect increased from 6% to 42% by CMD, from 12% to 54% by CEZ, from 20% to 58% by IPM/CS and from 78% to 84% by MINO. The combination of CMZ and FOM was found to be the most effective for enhanced effects on MRSA in vitro.


Programmed assembly of DNA-coated nanowire devices.
Combining biomolecular function with integrated circuit technology could usher in a new era of biologically enabled electronics. A key challenge has been coupling different molecular functions to specific chip locations for communication with the circuit. We used spatially confined electric fields to assemble different populations of DNA-coated nanowires to desired positions with an accuracy that enabled postassembly fabrication of contacts to each individual nanowire, with high yield and without loss of DNA function. This combination of off-chip synthesis and biofunctionalization with high-density, heterogeneous assembly and integration at the individual nanowire level points to new ways of incorporating biological functionality with silicon electronics.


Associations of the Initial COVID-19 Lockdown on Self-Reported Happiness and Worry about Developing Loneliness: A Cross-Sectional Analysis of Rural, Regional, and Urban Australian Communities.
Australia adopted hard lockdown measures to eliminate community transmission of COVID-19. Lockdown imposes periods of social isolation that contributes to increased levels of stress, anxiety, depression, loneliness, and worry. We examined whether lockdowns have similar psychosocial associations across rural and urban areas and whether associations existed between happiness and worry of loneliness in the initial wave of the COVID-19 pandemic in Australia. Data were collected using the "COVID-19 Living Survey" between 13 and 20 May 2020 by BehaviourWorks Australia at the Monash Sustainable Development Institute. The mean self-reported feeling of happiness and anxiousness (N = 1593), on a 10-point Likert scale with 0 being least happy or highly anxious, was 6.5 (SD = 2.4) and 3.9 (2.9), respectively. Factors associated with happiness were older age and having a postgraduate education. Participants worried about becoming lonely also exhibited reduced happiness (estimate = -1.58, 95%CI = -1.84--1.32) and higher anxiousness (2.22, 1.93-2.51) scores, and these conditions remained associated after adjusting for demographics. Interestingly, worry about loneliness was greater in rural areas than in urban communities. The negative impact of the COVID-19 lockdown on rural youth and those less-educated was evident. Participants in rural Australia who were worried about becoming lonely were reportedly less happy than participants in major cities. This dataset provides a better understanding of factors that influence psychological well-being and quality of life in the Australian population and helps to determine whether happiness may be an associative factor that could mitigate self-feelings of anxiety and worry about loneliness.


The preparation of large surface area lanthanum based perovskite supports for AuPt nanoparticles: tuning the glycerol oxidation reaction pathway by switching the perovskite B site.
Gold and gold alloys, in the form of supported nanoparticles, have been shown over the last three decades to be highly effective oxidation catalysts. Mixed metal oxide perovskites, with their high structural tolerance, are ideal for investigating how changes in the chemical composition of supports affect the catalysts' properties, while retaining similar surface areas, morphologies and metal co-ordinations. However, a significant disadvantage of using perovskites as supports is their high crystallinity and small surface area. We report the use of a supercritical carbon dioxide anti-solvent precipitation methodology to prepare large surface area lanthanum based perovskites, making the deposition of 1 wt% AuPt nanoparticles feasible. These catalysts were used for the selective oxidation of glycerol. By changing the elemental composition of the perovskite B site, we dramatically altered the reaction pathway between a sequential oxidation route to glyceric or tartronic acid and a dehydration reaction pathway to lactic acid. Selectivity profiles were correlated to reported oxygen adsorption capacities of the perovskite supports and also to changes in the AuPt nanoparticle morphologies. Extended time on line analysis using the best oxidation catalyst (AuPt/LaMnO3) produced an exceptionally high tartronic acid yield. LaMnO3 produced from alternative preparation methods was found to have lower activities, but gave comparable selectivity profiles to that produced using the supercritical carbon dioxide anti-solvent precipitation methodology.


Natural antioxidants in Alzheimer's disease.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterised by severe cognitive impairment that ultimately leads to death. Current drugs used in AD are acetylcholinesterase inhibitors and antagonists to the NMDA receptors. These drugs may only slightly improve cognitive functions but have only very limited impact on the clinical course of the disease. In the past several years, based on in vitro and in vivo studies in laboratory animals, natural antioxidants, such as resveratrol, curcumin and acetyl-L-carnitine have been proposed as alternative therapeutic agents for AD. An increasing number of studies demonstrated the efficacy of primary antioxidants, such as polyphenols, or secondary antioxidants, such as acetylcarnitine, to reduce or to block neuronal death occurring in the pathophysiology of this disorder. These studies revealed that other mechanisms than the antioxidant activities could be involved in the neuroprotective effect of these compounds. This paper discusses the evidence for the role of acetylcarnitine in modulating redox-dependent mechanisms leading to the upregulation of vitagenes. Furthermore, future development of novel antioxidant drugs targeted to the mitochondria should result in effectively slowing disease progression. The association with new drug delivery systems may be desirable and useful for the therapeutic use of antioxidants in human neurodegenerative diseases.


Polymeric mannosides prevent DC-SIGN-mediated cell-infection by cytomegalovirus.
Human cytomegalovirus (HCMV) is a beta-herpesvirus with a high prevalence in the population. HCMV is asymptomatic for immunocompetent adults but is a leading cause of morbidity for new born and immunocompromised patients. It was recently shown that the envelope glycoprotein B (gB) of HCMV interacts with the Dendritic Cell-Specific ICAM-3 Grabbing Non integrin (DC-SIGN) to infect the host. In this work we developed a set of DC-SIGN blockers based on mono-, di-, tetra and polyvalent mannosides. The multivalent mannosides were designed to interact with the carbohydrate recognition domains of DC-SIGN in a chelate or bind and recapture process, and represent the first chemical antiadhesives of HCMV reported so far. Polymeric dextrans coated with triazolylheptylmannoside (THM) ligands were highly potent, blocking the gB and DC-SIGN interaction at nanomolar concentrations. The compounds were further assessed for their ability to prevent the DC-SIGN mediated HCMV infection of dendritic cells. A dextran polymer coated with an average of 902 THM ligands showed an outstanding effect in blocking the HCMV trans-infection with IC50 values down to the picomolar range (nanomolar when expressed in THM concentration). Each THM moiety on the polymer surpassed the antiadhesive effect of the methylmannoside reference by more than four orders of magnitude. The compound proved non-cytotoxic at the high concentration of 2 mM and therefore represents an interesting antiadhesive candidate against HCMV and potentially against other virus hijacking dendritic cells to infect the host.


Neurochemistry of major depression: a study using magnetic resonance spectroscopy.
Magnetic resonance spectroscopy (MRS) is an acceptable non-invasive means of studying brain neurochemistry in depression. Previous studies in depressed patients have focused on measurement of the amino acid neurotransmitters, γ-aminobutyric acid (GABA) and glutamate. The aim of this study is to use MRS in conjunction with the ultrashort echo time 'SPECIAL' technique to measure cortical levels of GABA, glutamate and glutathione (GSH) levels in unmedicated patients with major depression. We also examined the effect of 6-week treatment with the selective serotonin re-uptake inhibitor, escitalopram. We studied patients with DSM-IV major depression and healthy age-matched controls using proton MRS. GABA, glutamate and GSH were measured relative to creatine in a voxel placed in occipital cortex. There was no difference in GABA or glutamate levels between depressed participants and controls; however, depressed patients had lower GSH levels. Six-week escitalopram treatment, which resulted in significant clinical responses in some patients, did not alter concentrations of GABA, glutamate or GSH. The sources of variability of GABA and glutamate measures in different studies of depressed patients require further study. Our results suggest that concomitant treatment with selective serotonin re-uptake inhibitors (SSRIs) is unlikely to be an important confounding factor. If lowered GSH levels can be confirmed, they may represent the presence of oxidative stress in some depressed patients.


Acetylcholinesterase inhibitors combined with memantine for moderate to severe Alzheimer's disease: a meta-analysis.
The clinical efficacy and safety of combination therapy with acetylcholinesterase inhibitor (AChEI) and memantine compared to AChEI or memantine alone in patients with Alzheimer's disease is inconclusive. We conducted a systematic review and meta-analysis of randomised controlled trials (RCTs) comparing the clinical efficacy and safety of combination therapy of AChEI and memantine to monotherapy with either substance in patients with moderate to severe Alzheimer's disease (Mini-Mental State Examination score is <20). We systematically searched EMBASE, Medline and CENTRAL until February 2018 for eligible RCTs. We pooled the outcome data using inverse variance weighting models assuming random effects, and assessed the quality of evidence (QoE) according to the Grading of Recommendations Assessment, Development and Evaluation (GRADE). We included nine RCTs (2604 patients). At short-term follow-up (closest to 6 months), combination therapy compared to AChEI monotherapy had a significantly greater effect on cognition than AChEI monotherapy (standardised mean difference [SMD] 0.20, 95% confidence interval [CI] 0.05 to 0.35, 7 RCTs, low QoE) and clinical global impression (SMD −0.15, 95% CI −0.28 to −0.01, 4 RCTs, moderate QoE), but not on activities of daily living (SMD 0.09, 95% CI −0.01 to 0.18, 5 RCTs, moderate QoE) or behavioural and psychological symptoms of dementia (mean difference −3.07, 95% CI −6.53 to 0.38, 6 RCT, low QoE). There was no significant difference in adverse events (relative risk ratio 1.05, 95% CI 0.98 to 1.12, 4 RCTs, low QoE). Evidence for long-term follow-up (≥ 9 months) or nursing home placement was sparse. Only two studies compared combination therapy with memantine monotherapy. Combination therapy had statistically significant effects on cognition and clinical global impression. The clinical relevance of these effects is uncertain. The overall QoE was very low. With the current evidence, it remains unclear whether combination therapy adds any benefit. Large pragmatic RCTs with long-term follow-up and focus on functional outcomes, delay in nursing home placement and adverse events are needed. &nbsp.


First Report of Grapevine leafroll-associated virus 2 in a Hybrid Grape 'Vidal Blanc' in Missouri.
Grapevine leaf roll disease (GLRD) is one of the most prevalent viral diseases in vineyards worldwide. At least 10 Grapevine leafroll-associated viruses (GLRaV-1 to -7, and -9, -10, and -11) are associated with GLRD. GLRaV-2 has a number of distinct isolates that are associated not only with GLRD, but also with graft incompatibility, young vine decline, and rootstock stem lesions. In the United States, GLRaV-2 isolates have been reported in the states of California (4), New York (1), Oregon, and Washington (3). A survey of grapevine viruses was conducted on seven grape cultivars, Norton, Chambourcin, Chardonel, Vignoles, Vidal Blanc, Traminette, and Cayuga White, in a vineyard located at Mountain Grove, MO during the falls of 2009 and 2010 and late summer of 2011. Vines from each cultivar were assigned into three blocks. From each block, six individual vines were randomly selected. Phloem scrapings were collected for detecting viruses by ELISA and reverse-transcription (RT)-PCR. To test for GLRaV-2, a set of primers, GLRaV-2-Forward (5'-GGTGATAACCGACGCCTCTA, nt 6745 to 6764) and GLRaV-2-Reverse (5'-CCTAGCTGACGCAGATTGCT, nt 7268 to 7287), were designed from the coat protein coding region as reported previously (2). GRLaV-2 was detected only in the samples of 'Vidal Blanc'. The detection of GLRaV-2 was further verified by ELISA using GRLaV-2-specific antisera (Bioreba AG, Reinach, Switzerland). The GLRaV-2-infected 'Vidal Blanc' vines did not exhibit visible symptoms. The RT-PCR amplified 543-bp cDNA fragments were isolated from agarose gel and cloned into the pCR2.1 vector (Invitrogen, Carlsbad, CA). Plasmid DNA was purified from three individual clones and sequenced from both directions. Comparison of the 543-bp sequences showed that the sequences shared 99% nt identity with the corresponding regions of 21 GLRaV-2 isolates, including PMC-083 isolate from Croatia (GenBank Accession No. HM185277), LN isolate from China (GenBank No. FJ786017), SE isolate from Brazil (GenBank No. EU204909), and four isolates from Washington (GenBank Nos. EU760836, 760838, 760843, and 760848). 'Vidal Blanc' is a white interspecific hybrid grape. Absence of visible symptoms on this cultivar emphasizes the importance of planting certified grapevines that have been indexed for viruses. Discovery of GLRaV-2 in asymptomatic white grape cultivars will warn growers on the potential damages and epidemics of GLRD in Midwest vineyards and promote planting clean grapevines. To our knowledge, this is the first report of GLRaV-2 in grapevines in Missouri. References: (1) M. Fuchs et al. Plant Dis. 93:395, 2009. (2) G. Gambino and I. Gribaudo. Phytopathology 96:1223, 2006. (3) R. R. Martin et al. Plant Dis. 89:763, 2005. (4) A. Rowhani et al. Phytopathology (Abstr.) 92:(suppl.):S71, 2002.


Analgesia of Patients with Multiple Rib Fractures in Critical Care: A Survey of Healthcare Professionals in the UK.
Good analgesia has been shown to reduce the risk of pneumonia, chronic pain, and mortality in patients with multiple rib fractures (MRFs). This survey explores the current analgesic practice in the UK, protocol use, barriers to provision, and physician preferences. A web-based survey was distributed nationally to an enriched cohort of clinicians working in UK trauma units with an interest in MRF management. Seventy-nine healthcare professionals responded. A third (31.4%) reported that their department had a rib fracture pain protocol, 52.9% did not, and 15.7% were unsure. Significantly more respondents reported adequate pain control when a hospital protocol was present compared to when not (χ 2, p < 0.01). Inadequate analgesia, a poor cough, and inability to breathe deeply were the commonest complications reported by 81.4, 78.6, and 65.7%, respectively. Patient-controlled analgesia (PCA) was the most commonly used form of analgesia (38.6%) followed by thoracic epidural (TEA) (30.0%) and continuous opioid infusion (18.6%). However, TEA was the preferred method of analgesia among respondents (37.1%) followed by serratus block (21.4%), paravertebral block (17.1%), and PCA (14.3%). There is considerable variation among physicians in their current use of analgesic modalities, with opiate-based methods predominating despite a physician preference for regional techniques. Thoracic epidurals are preferred by physicians but of limited use as a result of contraindications, time pressures, and staff skill mix. Pain control is reported to be better handled when protocols are present. Further research focusing on currently utilized regional techniques is required in order to produce a validated standardized national protocol that is informed by the current practice, the evidence base, and limitations to service provision. There is considerable variation among physicians in their current use of analgesic modalities. Opiate-based methods dominate for thoracic trauma despite a physician preference for regional techniques, which can be challenging in this cohort due to contraindications, staff skill mix, and time pressures. Inadequate analgesia is common but is better managed when pain management protocols are available. Beard L, Holt B, Snelson C, Parcha C, Smith FG, Veenith T. Analgesia of Patients with Multiple Rib Fractures in Critical Care: A Survey of Healthcare Professionals in the UK. Indian J Crit Care Med 2020;24(3):184-189.


High-Cost Patients and Preventable Spending: A Population-Based Study.
Although high-cost (HC) patients make up a small proportion of patients, they account for most health system costs. However, little is known about HC patients with cancer or whether some of their care could potentially be prevented. This analysis sought to characterize HC patients with cancer and quantify the costs of preventable acute care (emergency department visits and inpatient hospitalizations). This analysis examined a population-based sample of all HC patients in Ontario in 2013. HC patients were defined as those above the 90th percentile of the cost distribution; all other patients were defined as non-high-cost (NHC). Patients with cancer were identified through the Ontario Cancer Registry. Sociodemographic and clinical characteristics were examined and the costs of preventable acute care for both groups by category of visit/condition were estimated using validated algorithms. Compared with NHC patients with cancer (n=369,422), HC patients with cancer (n=187,770) were older (mean age 70 vs 65 years), more likely to live in low-income neighborhoods (19% vs 16%), sicker, and more likely to live in long-term care homes (8% vs 0%). Although most patients from both cohorts tended to be diagnosed with breast, prostate, or colorectal cancer, those with multiple myeloma or pancreatic or liver cancers were overrepresented among the HC group. Moreover, HC patients were more likely to have advanced cancer at diagnosis and be in the initial or terminal phase of treatment compared with NHC patients. Among HC patients with cancer, 9% of spending stemmed from potentially preventable/avoidable acute care, whereas for NHC patients, this spending was approximately 30%. HC patients with cancer are a unique subpopulation. Given the type of care they receive, there seems to be limited scope to prevent acute care spending among this patient group. To reduce costs, other strategies, such as making hospital care more efficient and generating less costly encounters involving chemotherapy, should be explored.