The use of audit to set up a thrombolysis programme in the accident and emergency department.
To improve the thrombolysis service offered by Gloucester Royal Hospital, by reducing the "door to needle time" (DTN) to 30 min (from a median of 110 min), and increasing the proportion of patients with acute myocardial infarctions receiving thrombolysis to 70% (from 58%). This would be achieved by moving the thrombolysis programme from the coronary care unit (CCU) to the accident and emergency (A&E) department. The process of audit was used to identify an area of poor performance, set standards, acquire funding, demonstrate achievement, and subsequently secure recurrent funding. Gloucester Royal Hospital. 946 patients presenting consecutively to the A&E department with non-traumatic chest pain between August 1993 and March 1994. DTN, overall delay time, and acute myocardial infarction thrombolysis rate. 946 patients were assessed over the eight month period, of whom 266 (28%) had suffered an acute myocardial infarction; 182 (68%) received thrombolysis (compared to 58% previously P < 0.05). Median DTN was reduced to 38 min (v 110 min previously, P < 0.0006). 127 (70%) patients received thrombolysis in the A&E department, and 55 (30%) in the CCU. Median overall delay time between onset of pain and thrombolysis was 3 h 35 min; 70% of patients received thrombolysis within 6 h of onset of symptoms and 90% within 12 h. Re-audit has subsequently shown maintenance of improvement. An in-house A&E based thrombolysis programme works in the District General Hospital setting; the process of audit can be used to acquire, and subsequently secure, funding for the project. The key to successful implementation of change is sensible resource allocation into adequate staffing and appropriate education.


Three options for citation tracking: Google Scholar, Scopus and Web of Science.
Researchers turn to citation tracking to find the most influential articles for a particular topic and to see how often their own published papers are cited. For years researchers looking for this type of information had only one resource to consult: the Web of Science from Thomson Scientific. In 2004 two competitors emerged--Scopus from Elsevier and Google Scholar from Google. The research reported here uses citation analysis in an observational study examining these three databases; comparing citation counts for articles from two disciplines (oncology and condensed matter physics) and two years (1993 and 2003) to test the hypothesis that the different scholarly publication coverage provided by the three search tools will lead to different citation counts from each. Eleven journal titles with varying impact factors were selected from each discipline (oncology and condensed matter physics) using the Journal Citation Reports (JCR). All articles published in the selected titles were retrieved for the years 1993 and 2003, and a stratified random sample of articles was chosen, resulting in four sets of articles. During the week of November 7-12, 2005, the citation counts for each research article were extracted from the three sources. The actual citing references for a subset of the articles published in 2003 were also gathered from each of the three sources. For oncology 1993 Web of Science returned the highest average number of citations, 45.3. Scopus returned the highest average number of citations (8.9) for oncology 2003. Web of Science returned the highest number of citations for condensed matter physics 1993 and 2003 (22.5 and 3.9 respectively). The data showed a significant difference in the mean citation rates between all pairs of resources except between Google Scholar and Scopus for condensed matter physics 2003. For articles published in 2003 Google Scholar returned the largest amount of unique citing material for oncology and Web of Science returned the most for condensed matter physics. This study did not identify any one of these three resources as the answer to all citation tracking needs. Scopus showed strength in providing citing literature for current (2003) oncology articles, while Web of Science produced more citing material for 2003 and 1993 condensed matter physics, and 1993 oncology articles. All three tools returned some unique material. Our data indicate that the question of which tool provides the most complete set of citing literature may depend on the subject and publication year of a given article.


Temporary external fixation is safe in a combat environment.
External fixation has been used extensively during recent wars as a damage control measure for fractures in coalition forces being evacuated. We hypothesize that external fixation is a safe and effective initial stabilization procedure for combat-related open fractures. Records on 55 consecutive type III tibia fractures between March 2003 and September 2007 were reviewed. We stratified the complications related to external fixation as major, potential, and minor complications. We defined major complications as neurovascular injury, mechanical failure, septic joint, and pin tract osteomyelitis. Potential complications were defined as pins within 1 inch of the fracture, pin overpenetration (> or = 26 mm), pin without cortical purchase, and intracapsular pin placement. Minor complications were defined as pin tract infections, addition of pins or bars, and pin overpenetration (9-25 mm). "Successful application" was defined as the absence of major or potential complications. We recorded no major complications. There were 12 of 53 (22.6%) constructs and 21 of 228 (9.2%) pins inserted with potential complications. We detected minor complications in 27 of 53 (50.9%) constructs and 35 of 228 (15.3%) pins inserted; 41 of 53 (77.4%) constructs had no major or potential complications. Treatment of combat-related open tibia fractures with external fixation was 77% successful in our series. We recorded no major complications but demonstrated the possibility for technical improvement in one of the five constructs with potential complications. Despite the recorded potential and minor complications, external fixation is safe and effective as a temporary damage control in open fractures sustained in combat.


Interactions within and between parallel parietal-frontal networks involved in complex motor behaviors in prosimian galagos and a squirrel monkey.
Long-train intracortical microstimulation (ICMS) of motor (M1) and posterior parietal cortices (PPC) in primates reveals cortical domains for different ethologically relevant behaviors. How functional domains interact with each other in producing motor behaviors is not known. In this study, we tested our hypothesis that matching domains interact to produce a specific complex movement, whereas connections between nonmatching domains are involved in suppression of conflicting motor outputs to prevent competing movements. In anesthetized galagos, we used 500-ms trains of ICMS to evoke complex movements from a functional domain in M1 or PPC while simultaneously stimulating another mismatched or matched domain. We considered movements of different and similar directions evoked from chosen cortical sites distant or close to each other. Their trajectories and speeds were analyzed and compared with those evoked by simultaneous stimulation. Stimulation of two sites evoking same or complementary movements produced a similar but more pronounced movement or a combined movement, respectively. Stimulation of two sites representing movements of different directions resulted in partial or total suppression of one of these movements. Thus interactions between domains in M1 and PPC were additive when they were functionally matched across fields or antagonistic between functionally conflicting domains, especially in PPC, suggesting that mismatched domains are involved in mutual suppression. Simultaneous stimulation of unrelated domains (forelimb and face) produced both movements independently. Movements produced by the simultaneous stimulation of sites in domains of two cerebral hemispheres were largely independent, but some interactions were observed.NEW & NOTEWORTHY Long trains of electrical pulses applied simultaneously to two sites in motor cortical areas (M1, PPC) have shown that interactions of functionally matched domains (evoking similar movements) within these areas were additive to produce a specific complex movement. Interactions between functionally mismatched domains (evoking different movements) were mostly antagonistic, suggesting their involvement in mutual suppression of conflicting motor outputs to prevent competing movements. Simultaneous stimulation of unrelated domains (forelimb and face) produced both movements independently.


Risk ratios for contagious outcomes.
Epidemiologists commonly use the risk ratio to summarize the relationship between a binary covariate and outcome, even when outcomes may be dependent. Investigations of transmissible diseases in clusters-households, villages or small groups-often report risk ratios. Epidemiologists have warned that risk ratios may be misleading when outcomes are contagious, but the nature of this error is poorly understood. In this study, we assess the meaning of the risk ratio when outcomes are contagious. We provide a mathematical definition of infectious disease transmission within clusters, based on the canonical stochastic susceptible-infective model. From this characterization, we define the individual-level ratio of instantaneous infection risks as the inferential target, and evaluate the properties of the risk ratio as an approximation of this quantity. We exhibit analytically and by simulation the circumstances under which the risk ratio implies an effect whose direction is opposite that of the true effect of the covariate. In particular, the risk ratio can be greater than one even when the covariate reduces both individual-level susceptibility to infection, and transmissibility once infected. We explain these findings in the epidemiologic language of confounding and Simpson's paradox, underscoring the pitfalls of failing to account for transmission when outcomes are contagious.


A new audience segmentation tool for African Americans: the black identity classification scale.
Many health communications target African Americans in an attempt to remediate race-based health disparities. Such materials often assume that African Americans are culturally homogeneous; however, research indicates that African Americans are heterogeneous in their attitudes, behaviors, and beliefs. The Black Identity Classification Scale (BICS) was designed as a telephone-administered tool to segment African American audiences into 16 ethnic identity types. The BICS was pretested using focus groups, telephone pretests, and a pilot study (n = 306). The final scale then was administered to 625 Black adults participating in a dietary intervention study, where it generally demonstrated good internal consistency reliability. The construct validity of the BICS also was explored by comparing participants' responses to culturally associated survey items. The distribution of the 16 BICS identity types in the intervention study is presented, as well as select characteristics for participants with core identity components. Although additional research is warranted, these findings suggest that the BICS has good psychometric properties and may be an effective tool for identifying African American audience segments.


Alterations of single molecule fluorescence lifetimes in near-field optical microscopy.
Fluorescence lifetimes of single Rhodamine 6G molecules on silica surfaces were measured with pulsed laser excitation, time-correlated single photon counting, and near-field scanning optical microscopy (NSOM). The fluorescence lifetime varies with the position of a molecule relative to a near-field probe. Qualitative features of lifetime decreases are consistent with molecular excited state quenching effects near metal surfaces. The technique of NSOM provides a means of altering the environment of a single fluorescent molecule and its decay kinetics in a repeatable fashion.


The Allometry of Sound Frequency Bandwidth in Songbirds.
AbstractTheory predicts that allometric constraints on sound production should be stronger for the lower frequencies of vocalizations than for the higher frequencies, which could originate from an allometry for sound frequency bandwidth. Using song recordings of approximately 1,000 passerine species (from >75% passerine genera), we show a significantly steeper allometry for the lower song frequencies than for the higher song frequencies, resulting in a positive allometry of frequency bandwidth: larger species can use wider bandwidths than smaller species. The bandwidth allometry exists in songbirds (oscines) but not in nonoscine passerines, indicating that it emerges from a combination of constraints to sound frequency production or transmission and the evolved behavior of oscines: unlike the narrow bandwidths of most nonoscine songs, the learned songs of oscines often use wide bandwidths that can be limited by both lower and upper constraints to sound frequency. This bandwidth allometry has implications for several research topics in acoustic communication.


Liver transplantation in cirrhotic patients over 60 years of age.
Although life expectancy in Spain is above seventy years, age over sixty is considered a relative contraindication for liver transplantation (LT) in most centers. The aim of this study was to assess the outcome of LT in patients over sixty years of age comparing them to patients under that age. From January 1992 to August 1995, 61 cirrhotic patients underwent LT at our institution; of them, 43 (group I) were younger than 60 years (mean +/- SEM: 51.9 +/- 0.9 years, range: 37-59) and 18 patients (group II) were 60 years or older (64.1 +/- 0.7, range: 60-71). Main pre-transplant variables (sex, etiology of liver disease, presence of hepatocarcinoma, Child-Pugh's score and renal function) were similar in both groups. The follow-up (median and range) for group I was 28 and 3-47 months, and for group II 16.5 and 3-48 months. Actuarial survival rates at one and four years post-LT were respectively 88.3% and 85.6% for group I, and 87.8% and 87.8% for the group II (p = n.s.). There were no differences between both groups regarding the incidence of rejection, major infections, neurologic complications, renal failure, pathological bone fractures, diabetes mellitus or hypertension. Nevertheless, cardiovascular complications were significantly more frequent in group II (p = 0.002) although they were not the cause of death. In conclusion, our results show that the outcome of LT in patients over sixty years old is comparable to that observed in patients under that age. LT should not be contraindicated on the only basis of an age greater than 60 years.


Liquids at large negative pressures: water at the homogeneous nucleation limit.
An isochoric cooling method for obtaining unprecedented tensions on liquids was used to determine the homogeneous nucleation limit for stretching of water at a variety of water densities. At densities in the range 0.55 to 0.68 gram per milliliter (g/ml), the data agree with the homogeneous nucleation temperatures measured by Skripov for superheated water at positive pressures. At densities between 0.68 and 0.93 g/ml, cavitation occurred only at negative pressures (that is, under tension). The cavitation tensions measured were in excellent agreement with those predicted by Fisher's 1948 vapor nucleation theory. A maximum tension of 140 megapascals (=1400 bars) was reached at 42 degrees C, which lies on an extrapolation of the line of isobaric density maxima. At higher densities, cavitation of droplets that survived heterogeneous nucleation failed to occur at all unless provoked, at much lower temperatures, by freezing. This observation confirms the existence of a density maximum at 42 degrees C and -140 megapascals and hence greatly strengthens the basis for Speedy's conjecture of a reentrant spinodal for water.


Enhanced liver regeneration following acute hepatectomy by low-level laser therapy.
The aim of the present study was to investigate the effect of low-level laser therapy (LLLT) on liver regeneration following hepatectomy. LLLT has been found to modulate various biological processes. Twelve mature male rats were used. The liver was exposed, and 70% of it was excised. The rats were assigned randomly to two groups: control, non-laser treated, and experimental, laser-treated (diode [Ga-Al-As] laser 804 nm) group. For determination of newly formed blood vessels and proliferating cells, 5-Bromo-2'deoxyuridine (BrdU) was injected intraperitoneally. The rats were sacrificed 2 d post hepatectomy, and histological sections from each liver were processed for analysis of new blood-vessel formation using BrdU immunostaining kit. Mesenchymal stem cells (MSCs) were assessed using c-kit immunostaining. BrdU-labeled cells were counted as for estimation of newly formed hepatic cells. It was found that the number of proliferating cells (BrdU positive cells) per area in the regenerating regions of the livers were significantly (p < 0.01) 2.6-fold higher in the laser-treated rats than in the control non-laser-treated rats. The density of the newly formed blood vessels and c-kit immunopositive cells in the regenerating area of the laser-treated livers was significantly (p < 0.01) 3.3- and 2.3-fold respectively higher than the control non-laser treated livers. It is concluded that LLLT following acute hepatectomy most probably stimulates a significant enhancement of liver regeneration conducive to both the formation of new hepatocytes and MSCs and angiogenesis in the regenerating liver.


Processing map and essential cleavage sites of the nonstructural polyprotein encoded by ORF1 of the feline calicivirus genome.
Feline calicivirus (FCV) nonstructural proteins are translated as part of a large polyprotein that undergoes autocatalytic processing by the virus-encoded 3C-like proteinase. In this study, we mapped three new cleavage sites (E(46)/A(47), E(331)/D(332), and E(685)/N(686)) recognized by the virus proteinase in the N-terminal part of the open reading frame 1 (ORF1) polyprotein to complete the processing map. Taken together with two sites we identified previously (E(960)/A(961) and E(1071)/S(1072)), the FCV ORF1 polyprotein contains five cleavage sites that define the borders of six proteins with calculated molecular masses of 5.6, 32, 38.9, 30.1, 12.7, and 75.7 kDa, which we designated p5.6, p32, p39 (NTPase), p30, p13 (VPg), and p76 (Pro-Pol), respectively. Mutagenesis of the E to A in each of these cleavage sites in an infectious FCV cDNA clone was lethal for the virus, indicating that these cleavages are essential in a productive virus infection. Mutagenesis of two cleavage sites (E(1345)/T(1346) and E(1419)/G(1420)) within the 75.7-kDa Pro-Pol protein previously mapped in bacterial expression studies was not lethal.


MicroRNAs are significantly influenced by p53 and radiation in HCT116 human colon carcinoma cells.
Ionizing radiation is genotoxic to the cell, and p53 is commonly considered to be a key regulator that controls gene expression responding to the genotoxity of radiation. The expression profiles of microRNAs (miRNAs), which are small non-coding RNAs regulating the translation of target mRNAs, were analyzed to determine whether any correlation exists between miRNA expression, radiation response, and/or p53. The miRNA profiles were analyzed by microarray containing 470 human miRNA probes in HCT116 human colon carcinoma cells and their p53-null derivative. Thirty-eight miRNAs among the 138 flagged human miRNAs were selected by fold-change analysis. The expression levels of these 38 miRNAs were changed more than two-fold, and a total of 12 miRNAs were significantly affected by p53, radiation, and the combination of both. All 12 miRNAs had expression patterns correlated to p53, while two miRNAs were affected by radiation or the combined action of radiation and p53. In bioinformatics studies, these miRNAs had p53-binding sites with scores higher than 85% in their upstream regions, and some of their target genes were found to be involved in genotoxic responses. In conclusion, we have identified miRNAs influenced significantly by p53 and/or radiation in the HCT116 human colon carcinoma cell line model, and these miRNAs may have important roles in the regulation of genes involved the cellular responses to radiation.


Geographic distribution and clonal diversity of Streptococcus pneumoniae serotype 1 isolates.
Serotype 1 pneumococci are a major cause of serious disease and have been associated with outbreaks but are rarely carried. The high attack rate and lack of coverage of this serotype by the heptavalent conjugate vaccine prompted the characterization of a geographically diverse collection of 166 serotype 1 isolates from recent cases of invasive disease. The isolates were resolved by multilocus sequence typing into 16 clones, which clustered into three major lineages with very different geographic distributions. Lineage A isolates were exclusively from Europe and North America, lineage B isolates were predominantly from Africa and Israel, and lineage C isolates were mainly from Chile. There was no clear association between the presence of individual clones within a country and the prevalence of serotype 1 disease.


A transgenic mouse model demonstrates a dominant negative effect of a point mutation in the RPS19 gene associated with Diamond-Blackfan anemia.
Diamond Blackfan anemia (DBA) is an inherited erythroblastopenia associated with mutations in at least 8 different ribosomal protein genes. Mutations in the gene encoding ribosomal protein S19 (RPS19) have been identified in approximately 25% of DBA families. Most of these mutations disrupt either the translation or stability of the RPS19 protein and are predicted to cause DBA by haploinsufficiency. However, approximately 30% of RPS19 mutations are missense mutations that do not alter the stability of the RPS19 protein and are hypothesized to act by a dominant negative mechanism. To formally test this hypothesis, we generated a transgenic mouse model expressing an RPS19 mutation in which an arginine residue is replaced with a tryptophan residue at codon 62 (RPS19R62W). Constitutive expression of RPS19R62W in developing mice was lethal. Conditional expression of RPS19R62W resulted in growth retardation, a mild anemia with reduced numbers of erythroid progenitors, and significant inhibition of terminal erythroid maturation, similar to DBA. RNA profiling demonstrated more than 700 dysregulated genes belonging to the same pathways that are disrupted in RNA profiles of DBA patient cells. We conclude that RPS19R62W is a dominant negative DBA mutation.


Pandemic potential of highly pathogenic avian influenza clade 220.127.116.11 A(H5) viruses.
The panzootic caused by A/goose/Guangdong/1/96-lineage highly pathogenic avian influenza (HPAI) A(H5) viruses has occurred in multiple waves since 1996. From 2013 onwards, clade 18.104.22.168 viruses of subtypes A(H5N2), A(H5N6), and A(H5N8) emerged to cause panzootic waves of unprecedented magnitude among avian species accompanied by severe losses to the poultry industry around the world. Clade 184.108.40.206 A(H5) viruses have expanded in distinct geographical and evolutionary pathways likely via long distance migratory bird dispersal onto several continents and by poultry trade among neighboring countries. Coupled with regional circulation, the viruses have evolved further by reassorting with local viruses. As of February 2019, there have been 23 cases of humans infected with clade 126.96.36.199 H5N6 viruses, 16 (70%) of which had fatal outcomes. To date, no HPAI A(H5) virus has caused sustainable human-to-human transmission. However, due to the lack of population immunity in humans and ongoing evolution of the virus, there is a continuing risk that clade 126.96.36.199 A(H5) viruses could cause an influenza pandemic if the ability to transmit efficiently among humans was gained. Therefore, multisectoral collaborations among the animal, environmental, and public health sectors are essential to conduct risk assessments and develop countermeasures to prevent disease and to control spread. In this article, we describe an assessment of the likelihood of clade 22.214.171.124 A(H5) viruses gaining human-to-human transmissibility and impact on human health should such human-to-human transmission occur. This structured analysis assessed properties of the virus, attributes of the human population, and ecology and epidemiology of these viruses in animal hosts.


Why Ecologists Should Care about Financial Markets.
Financial actors such as international banks and investors play an important role in the global economy. This role is shifting due to financial innovations, increased sustainability ambitions from large financial actors, and changes in international commodity markets. These changes are creating new global connections that potentially make financial markets, actors, and instruments important aspects of global environmental change. Despite this, the way financial markets and actors affect ecosystem change in different parts of the world has seldom been elaborated in the literature. We summarize these financial trends, explore how they connect to ecosystems and ecological change in both direct and indirect ways, and elaborate on crucial research gaps.


Pennsylvania Department of Corrections et al. v. Ronald R. Yeskey: prisons and the Americans with Disabilities Act of 1990.
This article reviews a recent U.S. Supreme Court decision that addresses the issue of whether Title II of the Americans with Disabilities Act of 1990, which prohibits a public entity from discriminating against a person with a disability due to that individual's disability, applies to inmates in state prisons. Potential ramifications of this important decision are briefly addressed.


Uganda experience-Using cost assessment of an established registry to project resources required to expand cancer registration.
The objectives of this study are (1) to estimate the cost of operating the Kampala Cancer Registry (KCR) and (2) to use cost data from the KCR to project the resource needs and cost of expanding and sustaining cancer registration in Uganda, focusing on the recently established Gulu Cancer Registry (GCR) in rural Northern Uganda. We used Centers for Disease Control and Prevention's (CDC's) International Registry Costing Tool (IntRegCosting Tool) to estimate the KCR's activity-based cost for 2014. We grouped the registry activities into fixed cost, variable core cost, and variable other cost activities. After a comparison KCR and GCR characteristics, we used the cost of the KCR to project the likely ongoing costs for the new GCR. The KCR incurred 42% of its expenditures in fixed cost activities, 40% for variable core cost activities, and the remaining 18% for variable other cost activities. The total cost per case registered was 28,201 Ugandan shillings (approximately US $10 in 2014) to collect and report cases using a combination of passive and active cancer data collection approaches. The GCR performs only active data collection, and covers a much larger area, but serves a smaller population compared to the KCR. After identifying many differences between KCR and GCR that could potentially affect the cost of registration, our best estimate is that the GCR, though newer and in a rural area, should require fewer resources than the KCR to sustain operations as a stand-alone entity. The optimal structure of the GCR needs to be determined in the future.


[Fluoride levels in saliva after tooth-brushing using fluoride toothpastes with and without rinsing of oral cavity].
The aim of the study was to determine salivary fluoride concentrations after tooth-brushing with fluoride toothpastes with and without rinsing of oral cavity. Fluoride levels in the supernatant of unstimulated mixed saliva were measured after tooth-brushing with Elmex (amine fluoride, 0.125% F) and Meridol (amine fluoride, stannous(II) fluoride, 0.14% F) toothpaste with and without rinsing. Fluoride concentration was measured using a fluoride ion-specific electrode (Orion 96 09BN) connected to a CPI-551 computer. The study was done in 120 subjects from whom salivary samples were taken before and after 15 and 30 min. from tooth-brushing with and without rinsing. Fluoride levels in saliva correlated with time from tooth-brushing and with oral cavity rinsing. Toothbrushing with rinsing led to similar increase/decrease in fluoride level in saliva for both toothpastes. The use of toothpaste without rinsing vs rinsing produced a two-fold increase in the level of salivary fluoride in the case of Elmex and a three-fold increase in the case of Meridol toothpaste. Fluoride content in saliva 30 minutes after brushing was higher than baseline. Significantly higher fluoride levels in saliva after tooth-brushing with fluoride toothpastes were noted when tooth-brushing was not followed by oral cavity rinsing.


Physicochemical and microbiological assessment of recreational and drinking waters.
The present study was aimed to make an assessment of health risk due to pollution and human pathogenic bacteria associated with the recreational and drinking water sources in twin densely populated holy Indian cities Ayodhya and Faizabad. Though physicochemical studies revealed that the water available in the area is under recommended limits for human use, it is unsafe on account of poor microbiological quality of surface and ground water in the region. The most probable number (MPN) test results revealed the preponderance of ≥2,400 total coliforms (TC) (100 ml)(-1) in river, pond, dug well and kund waters. Contrary to that, 94% tube wells, 32% hand pumps and 25% piped supply water were under safe limits having <3 TC (100 ml)(-1). The shallow depth (~40 ft), water logging and presence of septic tanks in the near vicinity are the possible reasons of poor microbial quality of hand pump drinking water. The municipal supply water passes along sewage line where loose connections and/or cracks in pipe lead to mixing and contamination. The significant best quality of tube well water evident from the absence of TC could be attributed to the depth of well ≥150 ft and usually their location away from the habitation. A total of 263 bacteria from 186 water samples were isolated, and at least five genera of enteric bacteria from various water sources were identified morphologically and biochemically as Escherichia coli, Klebsiella sp., Enterobacter sp., Shigella sp. and Salmonella sp. The serotyping of 72 E. coli and 36 Salmonella sp. revealed 51 as E. coli O157 and 20 as Salmonella sp. The presence of enteric pathogens in water sources pose threat to human health and therefore call for immediate remedial measures.


Environmentally responsive genome-wide accumulation of de novo Arabidopsis thaliana mutations and epimutations.
Evolution is fueled by phenotypic diversity, which is in turn due to underlying heritable genetic (and potentially epigenetic) variation. While environmental factors are well known to influence the accumulation of novel variation in microorganisms and human cancer cells, the extent to which the natural environment influences the accumulation of novel variation in plants is relatively unknown. Here we use whole-genome and whole-methylome sequencing to test if a specific environmental stress (high-salinity soil) changes the frequency and molecular profile of accumulated mutations and epimutations (changes in cytosine methylation status) in mutation accumulation (MA) lineages of Arabidopsis thaliana. We first show that stressed lineages accumulate ∼100% more mutations, and that these mutations exhibit a distinctive molecular mutational spectrum (specific increases in relative frequency of transversion and insertion/deletion [indel] mutations). We next show that stressed lineages accumulate ∼45% more differentially methylated cytosine positions (DMPs) at CG sites (CG-DMPs) than controls, and also show that while many (∼75%) of these CG-DMPs are inherited, some can be lost in subsequent generations. Finally, we show that stress-associated CG-DMPs arise more frequently in genic than in nongenic regions of the genome. We suggest that commonly encountered natural environmental stresses can accelerate the accumulation and change the profiles of novel inherited variants in plants. Our findings are significant because stress exposure is common among plants in the wild, and they suggest that environmental factors may significantly alter the rates and patterns of incidence of the inherited novel variants that fuel plant evolution.


Assessment of Resistance to Witches'-Broom Disease in Clonal and Segregating Populations of Theobroma cacao.
Fourteen clonal genotypes of cacao (Theobroma cacao) and their open-pollinated progenies were tested for their resistance to witches'-broom disease (Crinipellis perniciosa) using an agar-droplet inoculation method in separate, replicated greenhouse experiments. Nine measures of resistance were evaluated for precision, ability to discriminate between levels for resistance, and their correlation to field resistance. Correlations among the various measures of resistance were also ascertained. Data were re-examined to determine the accuracy and precision associated with individual plant measurement as well as the optimum sample size to assess segregating populations for resistance. The agar-droplet method was able to effectively discriminate various levels of resistance. Incubation period provided a precise, sensitive, valid, and heritable measure of resistance to C. perniciosa. Furthermore, incubation period was highly correlated with broom size and moderately correlated with broom frequency, two epidemiologically important components of resistance. The study also shows that 25 to 30 open-pollinated progeny are sufficient to provide a good estimate of a parent's resistance. The implications of the findings for identifying resistance among cacao germplasm collections as well as within segregating populations for resistance breeding are discussed.


Vitamin D for the prevention and treatment of pancreatic cancer.
Pancreatic cancer is ranked fifth among cancer-related deaths worldwide with a 5-year survival rate of less than 5%. Currently, surgery is the only effective therapy. However, most patients are diagnosed in the late stage and are not suitable for receiving curative surgery. Moreover, pancreatic cancer doesn't respond well to traditional chemotherapy and radiotherapy, leaving little effective treatment for advanced pancreatic cancer cases. 1alpha,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], the biologically active form of vitamin D(3), was originally identified during studies of calcium and bone metabolism, though it is now recognized that it exerts biological effects in almost every tissue in the body. Abundant evidence has shown that 1alpha,25(OH)(2)D(3) has antiproliferative, apoptotic, pro-differentiation and antiangiogensis effects in many types of cancer cells in vivo and in vitro, including breast, prostate, and colon. Similarly, the antitumor growth effect of 1alpha,25(OH)(2)D(3) on pancreatic cells has been demonstrated. The clinical use of 1alpha,25(OH)(2)D(3) is impeded by the lethal side effects of hypercalcemia and hypercalciuria. Therefore, 1alpha,25(OH)(2)D(3) analogs, which are either equipotent or more potent than 1alpha,25(OH)(2)D(3) in inhibiting tumor cell growth but with fewer hypercalcemic and hypercalciuric side effects, have been developed for the treatment of different cancers. Recently, a pre-clinical study demonstrated that a less calcemic analog of 1alpha,25(OH)(2)D(3), 19-nor-1alpha,25(OH)(2)D(2) (Paricalcitol), is effective in inhibiting tumor growth in vitro and in vivo, via upregulation of p21 and p27 tumor suppressor genes. Studies on the anti-tumor effects of a more potent analog of Paricalcitol are underway. 1alpha,25(OH)(2)D(3) and its analogs are potentially attractive novel therapies for pancreatic cancer.


Simultaneous modeling of antimycobacterial activities and ADMET profiles: a chemoinformatic approach to medicinal chemistry.
Mycobacteria represent a group of pathogens which cause serious diseases in mammals, including the lethal tuberculosis (Mycobacterium tuberculosis). Despite the mortality of this community-acquired and nosocomial disease mentioned above, other mycobacteria may cause similar infections, acting as dangerous opportunistic pathogens. Additionally, resistant strains belonging to Mycobacterium spp. have emerged. Thus, the design of novel antimycobacterial agents is a challenge for the scientific community. In this sense, chemoinformatics has played a vital role in drug discovery, helping to rationalize chemical synthesis, as well as the evaluation of pharmacological and ADMET (absorption, distribution, metabolism, excretion, toxicity) profiles in both medicinal and pharmaceutical chemistry. Until now, there is no in silico methodology able to assess antimycobacterial activity and ADMET properties at the same time. This work introduces the first multitasking model based on quantitative-structure biological effect relationships (mtk-QSBER) for simultaneous prediction of antimycobacterial activities and ADMET profiles of drugs/chemicals under diverse experimental conditions. The mtk-QSBER model was constructed by using a large and heterogeneous dataset of compounds (more than 34600 cases), displaying accuracies higher than 90% in both, training and prediction sets. To illustrate the utility of the present model, several molecular fragments were selected and their contributions to different biological effects were calculated and analyzed. Also, many properties of the investigational drug TMC-207 were predicted. Results confirmed that, from one side, TMC-207 can be a promising antimycobacterial drug, and on the other hand, this study demonstrates that the present mtk-QSBER model can be used for virtual screening of safer antimycobacterial agents.


A field-proven yeast two-hybrid protocol used to identify coronavirus-host protein-protein interactions.
Over the last 2 decades, yeast two-hybrid became an invaluable technique to decipher protein-protein interaction networks. In the field of virology, it has proven instrumental to identify virus-host interactions that are involved in viral embezzlement of cellular functions and inhibition of immune mechanisms. Here, we present a yeast two-hybrid protocol that has been used in our laboratory since 2006 to search for cellular partners of more than 300 viral proteins. Our aim was to develop a robust and straightforward pipeline, which minimizes false-positive interactions with a decent coverage of target cDNA libraries, and only requires a minimum of equipment. We also discuss reasons that motivated our technical choices and compromises that had to be made. This protocol has been used to screen most non-structural proteins of murine hepatitis virus (MHV), a member of betacoronavirus genus, against a mouse brain cDNA library. Typical results were obtained and are presented in this report.


Tula hantavirus triggers pro-apoptotic signals of ER stress in Vero E6 cells.
Tula virus is a member of the Hantavirus genus of the family Bunyaviridae. Viruses of this family have an unusual pattern of intracellular maturation at the ER-Golgi compartment. We recently found that Tula virus, similar to several other hantaviruses, is able to induce apoptosis in cultured cells [Li, X.D., Kukkonen, S., Vapalahti, O., Plyusnin, A., Lankinen, H., Vaheri, A., 2004. Tula hantavirus infection of Vero E6 cells induces apoptosis involving caspase 8 activation. J. Gen. Virol. 85, 3261-3268.]. However, the cellular mechanisms remain to be clarified. In this study, we demonstrate that the progressive replication of Tula virus in Vero E6 cells initiates several death programs that are intimately associated with ER stress: (1) early activation of ER-resident caspase-12; (2) phosphorylation of Jun NH2-terminal kinase (JNK) and its downstream target transcriptional factor, c-jun; (3) induction of the pro-apoptotic transcriptional factor, growth arrest- and DNA damage-inducible gene 153, or C/EBP homologous protein (Gadd153/chop); and (4) changes in the ER-membrane protein BAP31 implying cross-talk with the mitochondrial apoptosis pathway. Furthermore, we confirmed that a sustained ER stress was induced marked by an increased expression of an ER chaperone Grp78/BiP. Taken together, we have identified involvement of ER stress-mediated death program in Tula virus-infected Vero E6 cells which provides a new approach to understand the mechanisms in hantavirus-induced apoptosis.


Femtosecond surface plasmon interferometry.
We demonstrate femtosecond plasmonic interferometry with a novel geometry. The plasmonic microinterferometer consists of a tilted slit-groove pair. This arrangement allows for (i) interferometric measurements at a single wavelength with a single microinterferometer and (ii) unambiguous discrimination between changes in real and imaginary parts of the metal dielectric function. The performance is demonstrated by monitoring the sub-picosecond dynamics of hot electrons in gold.


Iliopsoas Hematoma in Patients Undergoing Venovenous ECMO.
Iliopsoas hematoma occasionally occurs in patients receiving anticoagulation therapy. It may be a life-threatening complication and can cause disseminated intravascular coagulation, which could develop into abdominal compartment syndrome. The incidence of and factors associated with iliopsoas hematoma during venovenous extracorporeal membrane oxygenation (ECMO) have not been well studied. To describe the incidence of iliopsoas hematoma and associated factors among patients undergoing venovenous ECMO. A retrospective cohort study was conducted at Nippon Medical School Hospital from April 2015 to October 2018. All patients (>18 years old) with iliopsoas hematoma received a diagnosis based on computed tomography. During the study period, 54 patients were supported with venovenous ECMO. Iliopsoas hematoma occurred in 8 of those patients (15%), none of whom had disseminated intravascular coagulopathy or abdominal compartment syndrome develop. Univariate analysis indicated that management of ECMO while the patient was awake and mobilization beyond sitting on the edge of the bed were significantly different (P < .05) in patients with and patients without iliopsoas hematoma. Mortality, however, did not differ significantly between the 2 groups. Our findings emphasize that recognizing factors associated with iliopsoas hematoma and detecting them early are crucial during venovenous ECMO in order to treat patients with iliopsoas hematoma appropriately.


Variation in Fourier transform infrared spectra of some homeopathic potencies and their diluent media.
The aim of this study was to determine whether potentized homeopathic drugs and their diluent media differ from each other with respect to their Fourier transform infrared (FTIR) spectra. FTIR spectra of Nux vomica 30C, Lycopodium 30C, Santonin 30C, Cina 30C, Cina 206C, Cina 1006C, and their diluent media 90% ethanol and Ethanol 30C were obtained in the wave number range of 2000-1000 cm1 at 20 degrees C. Potassium bromide powder soaked with the potencies, pressed into pellets, and air dried were used to measure the spectra. Because water structures in homeopathic potencies are thought to carry specific information on drug molecules and because O-H bending vibrational band (v2) exclusively belongs to water, the study was restricted to the bands in that wave number region. Alcohol has no absorption in the O-H bending region. The potencies were found to differ from each other and their diluent media in the number of v2 bands, their wave number (cm1), shape, and half-width (cm1) of the bands. The number and other characteristics of the v2 band represent the number of hydrogen-bonded water species and their hydrogen-bonding strength, respectively. The potencies and their diluent media therefore differ from each other in the number of hydrogen-bonded water species and their hydrogen-bonding strength. The observation that KBr pellets soaked with a potentized drug retains its specific spectral absorption properties simply confirms that medicated sucrose globules, used in homeopathic dispensing, are capable of retaining the therapeutic properties of the drug.


Bone marrow necrosis in antiphospholipid syndrome.
Bone marrow necrosis (BMN) is a relatively rare entity and has been associated with a poor prognosis. It is most commonly found in patients with neoplastic disorders, severe infections and sickle cell anemia. An unusual case of antiphospholipid syndrome (APS) with extensive bone marrow necrosis is described in a 27 year old woman. The patient presented with severe pancytopenia, some cognitive impairment resulting from a previous cerebrovascular accident, fever, hypertension, dyspnoea, tachycardia, hepatosplenomegaly, and vaginal bleeding. Her laboratory findings included a strongly positive Coombs' test (anti-IgG and anti-C3d), a prothrombin time of 23 seconds and an activated partial thromboplastin time of 45 seconds. Anticardiolipin antibody tests were positive. Antinuclear and anti-DNA antibodies were negative but the anti-SM test was positive. A bone marrow biopsy specimen was reported as showing extensive necrosis. The patient was treated with steroids, transfusion, and plasma exchange with some clinical improvement but her pancytopenia did not respond and necessitated frequent transfusions. This case lends further support to the association between APS and BMN.


Image-guided interventions and computer-integrated therapy: Quo vadis?
Significant efforts have been dedicated to minimizing invasiveness associated with surgical interventions, most of which have been possible thanks to the developments in medical imaging, surgical navigation, visualization and display technologies. Image-guided interventions have promised to dramatically change the way therapies are delivered to many organs. However, in spite of the development of many sophisticated technologies over the past two decades, other than some isolated examples of successful implementations, minimally invasive therapy is far from enjoying the wide acceptance once envisioned. This paper provides a large-scale overview of the state-of-the-art developments, identifies several barriers thought to have hampered the wider adoption of image-guided navigation, and suggests areas of research that may potentially advance the field.


Predictors for inpatient mortality during the first wave of the SARS-CoV-2 pandemic: A retrospective analysis.
The coronavirus disease 2019 (COVID-19) pandemic overwhelmed healthcare systems, highlighting the need to better understand predictors of mortality and the impact of medical interventions. This retrospective cohort study examined data from every patient who tested positive for COVID-19 and was admitted to White Plains Hospital between March 9, 2020, and June 3, 2020. We used binomial logistic regression to analyze data for all patients, and propensity score matching for those treated with hydroxychloroquine and convalescent plasma (CP). The primary outcome of interest was inpatient mortality. 1,108 admitted patients with COVID-19 were available for analysis, of which 124 (11.2%) were excluded due to incomplete data. Of the 984 patients included, 225 (22.9%) died. Risk for death decreased for each day later a patient was admitted [OR 0.970, CI 0.955 to 0.985; p < 0.001]. Elevated initial C-reactive protein (CRP) value was associated with a higher risk for death at 96 hours [OR 1.007, 1.002 to 1.012; p = 0.006]. Hydroxychloroquine and CP administration were each associated with increased mortality [OR 3.4, CI 1.614 to 7.396; p = 0.002, OR 2.8560, CI 1.361 to 6.160; p = 0.006 respectively]. Elevated CRP carried significant odds of early death. Hydroxychloroquine and CP were each associated with higher risk for death, although CP was without titers and was administered at a median of five days from admission. Randomized or controlled studies will better describe the impact of CP. Mortality decreased as the pandemic progressed, suggesting that institutional capacity for dynamic evaluation of process and outcome measures may benefit COVID-19 survival.


The Longitudinal Assessment of Imaging and Disability of the Back (LAIDBack) Study: baseline data.
Prospective cohort study of randomly selected Veterans Affairs (VA) outpatients. To determine the prevalence of magnetic resonance imaging (MRI) findings in the lumbar spine among persons without current low back pain or sciatica and to examine which findings are related to age or previous back symptoms. Previous studies of patients without low back pain have not explored the possible association of various MRI findings to past symptoms. We randomly selected an age-stratified sample of subjects without low back pain in the past 4 months from clinics at a VA hospital. We collected information on demographics, comorbidity, functional status, and quality of life. MR images were obtained using a standardized protocol through each of the five lumbar disc levels. Of 148 subjects, 69 (46%) had never experienced low back pain. There were 123 subjects (83%) with moderate to severe desiccation of one or more discs, 95 (64%) with one or more bulging discs, and 83 (56%) with loss of disc height. Forty-eight subjects (32%) had at least one disc protrusion and 9 (6%) had one or more disc extrusions. Many MR imaging findings have a high prevalence in subjects without low back pain. These findings are therefore of limited diagnostic use. The less common findings of moderate or severe central stenosis, root compression, and extrusions are likely to be diagnostically and clinically relevant.


Using sensor-based technology for safety and independence - the experiences of people with dementia and their families.
The majority of people with dementia prefer to live independently and safely in their own home cared for by their family members. Much effort has been invested in the development of technology, such as sensor-based networks. Many challenges remain, in particular gaining more knowledge about their experiences and perceived benefits. This study aimed to explore experiences, needs and benefits with using sensor-based technology for safety and independence in the homes of people with dementia and their family members. This study is part of the TECH@HOME project, aiming to evaluate the effects of sensor-based technology on independence among people with dementia and caregiver stress among their family members. This study applied an inductive, qualitative approach with semi-structured interviews of people with dementia (n = 9) and family members (n = 21). The participants were interviewed between June and September 2018 after using the technology for at least 6 months. The interviews were analysed with manifest content analysis. Our findings highlighted that technology was considered as a precaution and a safety measure that could provide a sense of having control of the everyday life of the person with dementia. Understanding and acceptance of the technology were as important, together with the reliability of the technology. Ethical dilemmas related to the monitoring of the person with dementia in the home were also raised. This study provides insights into how people with dementia and family members experience and benefit from using sensor-based technology in their own homes. The knowledge generated is essential for healthcare professionals and policymakers developing and implementing care and service systems including technology, as well as for the industry.


Within-group differences in depression among older Hispanics living in the United States.
Using the Health and Retirement Study, we examine the prevalence of depression in different groups of Hispanic older adults. Respondents (n = 759) were aged 59 and older and identified themselves as Mexican American (56%), Cuban American (13%), Puerto Rican (8%), other (8%), or not specified (15%). We used a modified version of the Center for Epidemiologic Studies-Depression scale and the Composite International Diagnostic Interview to assess depressive symptoms and the presence of major depression. Relative to Puerto Ricans, each Hispanic group had significantly lower levels of depressive symptoms, except for Cuban Americans; and each Hispanic group had lower prevalence rates for major depression, except for other Hispanics, even after we adjusted for sociodemographic, cultural factors, socioeconomic, functional limitations, and chronic health conditions.


Planarian MBD2/3 is required for adult stem cell pluripotency independently of DNA methylation.
Planarian adult stem cells (pASCs) or neoblasts represent an ideal system to study the evolution of stem cells and pluripotency as they underpin an unrivaled capacity for regeneration. We wish to understand the control of differentiation and pluripotency in pASCs and to understand how conserved, convergent or divergent these mechanisms are across the Bilateria. Here we show the planarian methyl-CpG Binding Domain 2/3 (mbd2/3) gene is required for pASC differentiation during regeneration and tissue homeostasis. The genome does not have detectable levels of 5-methylcytosine (5(m)C) and we find no role for a potential DNA methylase. We conclude that MBD proteins may have had an ancient role in broadly controlling animal stem cell pluripotency, but that DNA methylation is not involved in planarian stem cell differentiation.


Isolation and characterization of the human CP49 gene promoter.
This study focuses on the identification of regulatory elements that contribute to lens-specific expression of the human CP49 gene within the 5'-flanking DNA sequences. The DNA sequence upstream of the human CP49 coding region was subcloned as a set of 5' and 3' deletion series. The constructs were transfected into lens (N/N1003A) and nonlens (NIH3T3) cell lines and chicken primary lens cultures, to test for promoter activity and specificity. To further test the specificity, a portion of the 5' flanking DNA sequence was used to drive transgene expression in mice. The flanking DNA sequence was analyzed for potential transcription factor-binding sites. The 5'-flanking DNA preferentially activated reporter gene expression in a lens-preferred manner when transfected into cultured cells. Transgene expression driven by the CP49 promoter region was lens specific. Analysis of the proximal promoter sequence revealed the presence of potential binding sites for the AP-1, AP-2, and OCT-1 transcription factors and the absence of TATA and CAAT boxes. The sequence upstream of the CP49 gene possesses promoter activity and is able to drive lens-preferred expression in both transfection and transgenic experiments. Promoter activity is dependent on the presence of the proximal 300 bp directly upstream of the coding region.


Nucleostemin in injury-induced liver regeneration.
The high regenerative capacity of liver contributes to the maintenance of its size and function when injury occurs. Partial hepatectomy induces division of mature hepatocytes to maintain liver function, whereas severe injury stimulates expansion of undifferentiated hepatic precursor cells, which supply mature cells. Although several factors reportedly function in liver regeneration, the precise mechanisms underlying regeneration remain unclear. In this study, we analyzed expression of nucleostemin (NS) during development and in injured liver by using transgenic green fluorescent protein reporter (NS-GFP Tg) mice. In neonatal liver, the hepatic precursor cells that give rise to mature hepatocytes were enriched in a cell population expressing high levels of NS. In adult liver, NS was abundantly expressed in mature hepatocytes and rapidly upregulated by partial hepatectomy. Severe liver injury promoted by a diet containing 3,5-diethoxycarbonyl-1,4-dihydrocollidine induced the emergence of NS-expressing ductal epithelial cells as hepatic precursor cells. NS knockdown inhibited both hepatic colony formation in vitro and proliferation of hepatocytes in vivo. These data strongly suggest that NS plays a critical role in regeneration of both hepatic precursor cells and hepatocytes in response to liver injury.


A Third Class: Functional Gibberellin Biosynthetic Operon in Beta-Proteobacteria.
The ability of plant-associated microbes to produce gibberellin A (GA) phytohormones was first described for the fungal rice pathogen Gibberella fujikuroi in the 1930s. Recently the capacity to produce GAs was shown for several bacteria, including symbiotic alpha-proteobacteria (α-rhizobia) and gamma-proteobacteria phytopathogens. All necessary enzymes for GA production are encoded by a conserved operon, which appears to have undergone horizontal transfer between and within these two phylogenetic classes of bacteria. Here the operon was shown to be present and functional in a third class, the beta-proteobacteria, where it is found in several symbionts (β-rhizobia). Conservation of function was examined by biochemical characterization of the enzymes encoded by the operon from Paraburkholderia mimosarum LMG 23256T. Despite the in-frame gene fusion between the short-chain alcohol dehydrogenase/reductase and ferredoxin, the encoded enzymes exhibited the expected activity. Intriguingly, together these can only produce GA9, the immediate precursor to the bioactive GA4, as the cytochrome P450 (CYP115) that catalyzes the final hydroxylation reaction is missing, similar to most α-rhizobia. However, phylogenetic analysis indicates that the operon from β-rhizobia is more closely related to examples from gamma-proteobacteria, which almost invariably have CYP115 and, hence, can produce bioactive GA4. This indicates not only that β-rhizobia acquired the operon by horizontal gene transfer from gamma-proteobacteria, rather than α-rhizobia, but also that they independently lost CYP115 in parallel to the α-rhizobia, further hinting at the possibility of detrimental effects for the production of bioactive GA4 by these symbionts.


Acinetobacter pneumonia: Is the outcome different from the pneumonias caused by other agents.
The principal aim of the present study was to determine whether Acinetobacter spp. pneumonia differs from hospital-acquired pneumonias (HAPs) caused by other agents with respect to therapeutic success and survival rate. This study includes 140 adult patients diagnosed with HAPs caused by identified etiologic agents between March 2005 and February 2006. These patients were divided into two groups according to the agent responsible for their infection (Acinetobacter spp. [n = 63] or non-Acinetobacter spp. [n = 77]). The groups were compared in terms of risk factors, therapeutic success and six-week survival rates. Previous antibiotic use and the risk of aspiration were independent factors responsible for the development of Acinetobacter spp. pneumonia. Hypoalbuminemia, steroid use and the use of a mechanical ventilator were determined to be mortality-associated independent risk factors for Acinetobacter spp. pneumonia. The clinical success rate at the end of therapy was 41.6% and, at the sixth week, the survival rate was 35% among patients in whom Acinetobacter spp. was the causative agent. Conversely, in the control group, these values were 43 and 32%, respectively (P > 0.05). We found that the use of the appropriate antibiotics for the treatment of Acinetobacter spp. pneumonia was an important factor in survival (P < 0.001). The outcomes of Acinetobacter spp. pneumonia do not differ from HAPs associated with non-Acinetobacter spp. in terms of therapeutic success and survival rates.


The true cost of greenhouse gas emissions: analysis of 1,000 global companies.
This study elucidated the shadow price of greenhouse gas (GHG) emissions for 1,024 international companies worldwide that were surveyed from 15 industries in 37 major countries. Our results indicate that the shadow price of GHG at the firm level is much higher than indicated in previous studies. The higher shadow price was found in this study as a result of the use of Scope 3 GHG emissions data. The results of this research indicate that a firm would carry a high cost of GHG emissions if Scope 3 GHG emissions were the focus of the discussion of corporate social responsibility. In addition, such shadow prices were determined to differ substantially among countries, among sectors, and within sectors. Although a number of studies have calculated the shadow price of GHG emissions, these studies have employed country-level or industry-level data or a small sample of firm-level data in one country. This new data from a worldwide firm analysis of the shadow price of GHG emissions can play an important role in developing climate policy and promoting sustainable development.


Advances in surgical management of malignancies of the cranial base: the extended transbasal approach.
The extended transbasal approach combines a bifrontal craniotomy with an orbital nasal and potentially a sphenoethmoidal osteotomy to provide excellent access to malignancies of the anterior, middle and posterior skull base. The approach enables the en bloc resection of tumors within the frontal lobes, orbits, paranasal sinuses and sphenoclival corridors without brain retraction and may obviate the need for transfacial access. We present our 7-year experience during which 29 patients underwent surgery with the extended transbasal exposure. In 25 patients the extended transbasal approach was used alone; in the remaining four it was combined with additional approaches. With exception of two patients, all lesions were removed en bloc. Reconstruction was accomplished with the use of pericranium and in some instances a temporalis muscle pedicle or a gracilis microvascular free flap. There were no mortalities associated with this approach. Seven patients experienced infections, four patients experienced cerebral spinal fluid (CSF) leakage, two patients who had received adjuvant radiation experienced scalp necrosis, three patients experienced pneumocephalus, and 29 patients experienced cranial neuropathies, the majority of which were loss of olfaction. The average follow-up for our patients was 34 months with a range of 2--62 months.


Cultural Stress: The Ubiquitous Stressor Hiding in Plain Sight.
While most healthcare providers are generally aware of the role stress plays in disease development, few are attuned to a phenomenon I call "Cultural Stress," which compounds all of life's other stressors. Cultural Stress is the constant, pervasive, ever-increasing stress of modern living. It includes overexposure to technology at the expense of in-person relationships; 24-hour connectivity, which blurs the boundaries between work and personal life; on-demand delivery of goods and services, which severs neighborhood and community relationships and results in long sedentary hours in front of our screens; the pace of technological change, which contributes to professional, economic, and social insecurity; our global economic system, which sows uncertainty as it outsources jobs overseas or to new technologies; environmental contaminants, noise, crowding, and various new sources of radiation; changes to our diet, which has become heavily reliant on a few processed commodities (corn, wheat, sugar, and soy); and changes to our urban development and transportation patterns, which result in less physical activity and more hours spent sitting in long commutes. It is the chronic nature of Cultural Stress that distinguishes it from conventional stress. Because Cultural Stress is ubiquitous, the body seldom gets a chance to recover without active intervention strategies.


High thigh muscle strength but not bone mass in young horseback-riding females.
To evaluate whether the type of weight-bearing loading subjected to the skeleton during horseback-riding was associated with differences in bone mass and muscle strength of the thigh, we investigated bone mass and isokinetic muscle strength in 20 female horse riders (age 17.9 +/- 0.6 years) who were riding 7.0 +/- 3.4 hours/week, and 20 nonactive females (age 17.8 +/- 1.1 years). The groups were matched according to age, weight, and height. Areal bone mineral density was measured in total body, head, lumbar spine, right femoral neck, Ward's triangle, and trochanter, the whole dominant and nondominant humerus, and in specific sites in the right femur diaphysis, distal femur, proximal tibia, and tibia diaphysis using dual X-ray absorptiometry. Isokinetic concentric and eccentric peak torque of the quadricep and hamstring muscles were measured using an isokinetic dynamometer. There were no significant differences in bone mass between the horseback riders and nonactives at any site measured. The horse riders were significantly (P < 0.05-0.01) stronger in concentric hamstrings strength at 90 degrees/second and 225 degrees/second and in eccentric quadricep and hamstring strength at 90 degrees/second. Horseback riding in young females is associated with a high muscle strength of the thigh, but not with a high bone mass.


Computed tomography of dural sinus thrombosis.
For 150 years, the variable clinical appearance of dural sinus thrombosis has plagued clinicians. Computed tomography (CT) has alleviated much of the difficulty in making this diagnosis. A high density lesion in the involved sinus on precontrast scans and a filling defect in the sinus on postcontrast scans were the most frequently observed CT abnormalities in 12 patients with sinus thrombosis. Other findings include the "cord" sign and diffuse cerebral edema. Thrombosis of the superior sagittal sinus, transverse sinus, or sigmoid sinus can occur as an isolated event or in association with other disease entities and may cause no neurologic impairment.


Intestinal, but not hepatic, ChREBP is required for fructose tolerance.
Increased sugar consumption is a risk factor for the metabolic syndrome including obesity, hypertriglyceridemia, insulin resistance, diabetes, and nonalcoholic fatty liver disease (NAFLD). Carbohydrate responsive element-binding protein (ChREBP) is a transcription factor that responds to sugar consumption to regulate adaptive metabolic programs. Hepatic ChREBP is particularly responsive to fructose and global ChREBP-KO mice are intolerant to diets containing fructose. It has recently been suggested that ChREBP protects the liver from hepatotoxicity following high-fructose diets (HFrDs). We directly tested this hypothesis using tissue-specific ChREBP deletion. HFrD increased adiposity and impaired glucose homeostasis in control mice, responses that were prevented in liver-specific ChREBP-KO (LiChKO) mice. Moreover, LiChKO mice tolerated chronic HFrD without marked weight loss or hepatotoxicity. In contrast, intestine-specific ChREBP-KO (IChKO) mice rapidly lost weight after transition to HFrD, and this was associated with dilation of the small intestine and cecum, suggestive of malabsorption. These findings were associated with downregulation of the intestinal fructose transporter, Slc2a5, which is essential for fructose tolerance. Altogether, these results establish an essential role for intestinal, but not hepatic, ChREBP in fructose tolerance.


The Role of Early Maternal Support in Balancing Full-Time Work and Infant Exclusive Breastfeeding: A Qualitative Study.
Support of others is a key factor for mothers who choose to breastfeed their infants, including those who balance work outside the home and breastfeeding. However, little research has been done to understand how maternal support during the postpartum period impacts mothers' ability to later balance work and breastfeeding, in particular full-time work and exclusive breastfeeding. The results of this qualitative study indicate that the timing of support plays a key role in mothers' ability to successfully overcome barriers during the early postpartum period, thus building maternal self-efficacy in addressing problems encountered when they return to work. To understand the experience of low-income women who successfully balance full-time work and exclusive breastfeeding for the recommended 6 months, interviews were conducted with women who met study criteria for income level, work status, and exclusive breastfeeding. Breastfeeding peer counselors were also interviewed as key informants. Interviews were recorded, transcribed, and coded for themes. The results of both sets of interviews were triangulated with a focused literature review to assure the soundness of the qualitative analysis. Timing of support included acute support, such as help establishing a successful latch needed during the first 2 weeks after delivery, to deal with breastfeeding problems that mothers perceived as being mentally and emotionally overwhelming and longer-term support needed to overcome problems perceived as being less intense. The research invites further exploration into the relationship between breastfeeding support provided by mothers' support system, including healthcare professionals, during the postpartum period and rates of breastfeeding duration and exclusivity.


Effect of progressive resistive exercise training in improving mobility and functional ability of middle adulthood patients with chronic kidney disease.
Patients with chronic kidney disease (CKD) in their middle adulthood are more prone to reduced mobility than younger patients having the same medical condition. Progressive resistive exercise training (PRT) is deemed an effective treatment approach for the management of muscular weakness in patients with CKD. The present review is an attempt to understand the effectiveness of PRT in the mobility and functional ability of patients suffering from CKD. We systematically searched electronic databases, including Medline, Scopus, PubMed, CINAHL, PEDRo and Cochrane, to review the published literature on this subject. Electronic searches were limited to training programs carried out on resistive, aerobic, endurance and therapeutic exercises reporting outcome measures including muscular strength, size, physical function and functional capacity in the clinical population with CKD aged > 40 years. Studies with a minimum duration of eight weeks of exercise training or more were considered eligible for review. The methodological criteria of the included studies were assessed with the PEDro scale. A total of 80 articles were identified using the keywords in the above-mentioned databases. However, based on the study's inclusion and exclusion criteria, only 11 articles were finally included. The results of this review substantiate the effectiveness of PRT in patients with CKD. However, further research is warranted in this area due to the limited availability of high-quality published evidence.


Characterisation of pore structures of pharmaceutical tablets: A review.
Traditionally, the development of a new solid dosage form is formulation-driven and less focus is put on the design of a specific microstructure for the drug delivery system. However, the compaction process particularly impacts the microstructure, or more precisely, the pore architecture in a pharmaceutical tablet. Besides the formulation, the pore structure is a major contributor to the overall performance of oral solid dosage forms as it directly affects the liquid uptake rate, which is the very first step of the dissolution process. In future, additive manufacturing is a potential game changer to design the inner structures and realise a tailor-made pore structure. In pharmaceutical development the pore structure is most commonly only described by the total porosity of the tablet matrix. Yet it is of great importance to consider other parameters to fully resolve the interplay between microstructure and dosage form performance. Specifically, tortuosity, connectivity, as well as pore shape, size and orientation all impact the flow paths and play an important role in describing the fluid flow in a pharmaceutical tablet. This review presents the key properties of the pore structures in solid dosage forms and it discusses how to measure these properties. In particular, the principles, advantages and limitations of helium pycnometry, mercury porosimetry, terahertz time-domain spectroscopy, nuclear magnetic resonance and X-ray computed microtomography are discussed.


Niemann-Pick C1 protein facilitates the efflux of the anticancer drug daunorubicin from cells according to a novel vesicle-mediated pathway.
Niemann-Pick C1 (NPC1) is a late endosomal/lysosomal membrane protein originally reported on for its role in cholesterol trafficking in mammalian cells. NPC1 has been shown recently to share significant structural homology with a family of prokaryotic permeases and was proposed to play a role in intracellular drug transport; however, the mechanism for this has not been fully understood. We provide evidence here that is consistent with NPC1's involvement in a vesicle-mediated clearance of the anticancer agent daunorubicin from cells. In experiments with human fibroblasts, we demonstrate that lysosomal efflux of daunorubicin, as well as dextran molecules, are significantly reduced in cells with mutated and dysfunctional NPC1 compared with wild-type fibroblasts. Furthermore, we show that NPC1 is implicated in a lysosomal drug sequestration phenotype exhibited by the multidrug-resistant (MDR) human leukemic HL-60 cancer cell line. Evaluations of cholesterol trafficking, NPC1 mRNA levels, and protein expression are all consistent with a loss of NPC1 activity that is associated with the emergence of the MDR phenotype in this cell line. Collectively, this work proposes a novel role for NPC1 in a vesicle-mediated pathway responsible for the clearance of drugs from cells and provides an explanation for a drug sequestration phenotype exhibited by the MDR HL-60 cell line.


Psychometric properties of startle and corrugator response in NPU, affective picture viewing, and resting state tasks.
The current study provides a comprehensive evaluation of critical psychometric properties of commonly used psychophysiology laboratory tasks/measures within the NIMH RDoC. Participants (N = 128) completed the no-shock, predictable shock, unpredictable shock (NPU) task, affective picture viewing task, and resting state task at two study visits separated by 1 week. We examined potentiation/modulation scores in NPU (predictable or unpredictable shock vs. no-shock) and affective picture viewing tasks (pleasant or unpleasant vs. neutral pictures) for startle and corrugator responses with two commonly used quantification methods. We quantified startle potentiation/modulation scores with raw and standardized responses. We quantified corrugator potentiation/modulation in the time and frequency domains. We quantified general startle reactivity in the resting state task as the mean raw startle response during the task. For these three tasks, two measures, and two quantification methods, we evaluated effect size robustness and stability, internal consistency (i.e., split-half reliability), and 1-week temporal stability. The psychometric properties of startle potentiation in the NPU task were good, but concerns were noted for corrugator potentiation in this task. Some concerns also were noted for the psychometric properties of both startle and corrugator modulation in the affective picture viewing task, in particular, for pleasant picture modulation. Psychometric properties of general startle reactivity in the resting state task were good. Some salient differences in the psychometric properties of the NPU and affective picture viewing tasks were observed within and across quantification methods.


Novel biomarkers for the detection of prostate cancer.
Prostate-specific antigen (PSA) is widely used as a biomarker in the detection of prostate cancer and for decision making regarding treatment options, response to therapy, and clinical follow-up. Despite its widespread use, it is well recognised that PSA has suboptimal performance as a screening tool due to poor specificity, resulting in high negative biopsy rates and potential 'over-diagnosis' and 'over-treatment' of clinically insignificant cancers. In particular, PSA does not reliably distinguish either cancer from benign prostatic conditions, or 'clinically significant' from 'indolent cancers', and it is inaccurate in predicting disease burden and response to treatment. There is an urgent demand for novel biomarkers to address these clinical needs. This article provides an update on the novel candidate biomarkers in development, which have shown potential for improving the detection of clinically significant cases of this malignancy.


The Bloom's syndrome helicase (BLM) interacts physically and functionally with p12, the smallest subunit of human DNA polymerase delta.
Bloom's syndrome (BS) is a cancer predisposition disorder caused by mutation of the BLM gene, encoding a member of the RecQ helicase family. Although the phenotype of BS cells is suggestive of a role for BLM in repair of stalled or damaged replication forks, thus far there has been no direct evidence that BLM associates with any of the three human replicative DNA polymerases. Here, we show that BLM interacts specifically in vitro and in vivo with p12, the smallest subunit of human POL delta (hPOL delta). The hPOL delta enzyme, as well as the isolated p12 subunit, stimulates the DNA helicase activity of BLM. Conversely, BLM stimulates hPOL delta strand displacement activity. Our results provide the first functional link between BLM and the replicative machinery in human cells, and suggest that BLM might be recruited to sites of disrupted replication through an interaction with hPOL delta. Finally, our data also define a novel role for the poorly characterized p12 subunit of hPOL delta.


The future of novel diagnostics in medical mycology.
Several fungal diseases have become serious threats to human health and life, especially upon the advent of human immunodeficiency virus/AIDS epidemics and of other typical immunosuppressive conditions of modern life. Accordingly, the burden posed by these diseases and, concurrently, by intensive therapeutic regimens against these diseases has increased worldwide. Existing and available rapid tests for point-of-care diagnosis of important fungal diseases could enable the limitations of current laboratory methods for detection and identification of medically important fungi to be surpassed, both in low-income countries and for first-line diagnosis (screening) in richer countries. As with conventional diagnostic methods and devices, former immunodiagnostics have been challenged by molecular biology-based platforms, as a way to enhance the sensitivity and shorten the assay time, thus enabling early and more accurate diagnosis. Most of these tests have been developed in-house, without adequate validation and standardization. Another challenge has been the DNA extraction step, which is especially critical when dealing with fungi. In this paper, we have identified three major research trends in this field: (1) the application of newer biorecognition techniques, often applied in analytical chemistry; (2) the development of new materials with improved physico-chemical properties; and (3) novel bioanalytical platforms, allowing fully automated testing. Keeping up to date with the fast technological advances registered in this field, primarily at the proof-of-concept level, is essential for wise assessment of those that are likely to be more cost effective and, as already observed for bacterial and viral pathogens, may provide leverage to the current tepid developmental status of novel and improved diagnostics for medical mycology.


High prevalence of liver and intestinal fluke infections among residents of Savannakhet Province in Laos.
The prevalence of liver and intestinal fluke infections was surveyed on residents of Savannakhet Province, Laos. Fecal specimens were collected from a total of 981 residents in 4 Mekong riverside villages and examined by the Kato-Katz thick smear technique. The results revealed that the overall helminth egg positive rate was 84.2%, and the positive rate for small trematode eggs, including Opisthorchis viverrini, heterophyids, or lecithodendriids, was 67.1%. To obtain adult flukes, 38 small trematode egg positive cases were treated with a 20-30 mg/kg single dose of praziquantel and purged. Diarrheic stools were then collected from 29 people and searched for helminth parasites using stereomicroscopes. Mixed infections with O. viverrini and 6 kinds of intestinal flukes were found, namely, Haplorchis taichui, Haplorchis pumilio, Haplorchis yokogawai, Prosthodendrium molenkampi, Phaneropsolus bonnei, and echinostomes. The total number of flukes collected was 7,693 specimens (av. no. per treated person; 265.3). The most common species was O. viverrini, followed by H. taichui, P. molenkampi, echinostomes, H. pumilio, P. bonnei, and H. yokogawai. The results indicate that foodborne liver and intestinal fluke infections are prevalent among residents of Savannakhet Province, Laos.


Memory-like innate response to booster vaccination with MF-59 adjuvanted influenza vaccine in children.
The pediatric population receives the majority of vaccines globally, yet there is a paucity of studies on the transcriptional response induced by immunization in this special population. In this study, we performed a systems-level analysis of immune responses to the trivalent inactivated influenza vaccine adjuvanted with MF-59 in children (15-24 months old) and in young, healthy adults. We analyzed transcriptional responses elicited by vaccination in peripheral blood, as well as cellular and antibody responses following primary and booster vaccinations. Our analysis revealed that primary vaccination induced a persistent transcriptional signature of innate immunity; booster vaccination induced a transcriptional signature of an enhanced memory-like innate response, which was consistent with enhanced activation of myeloid cells assessed by flow cytometry. Furthermore, we identified a transcriptional signature of type 1 interferon response post-booster vaccination and at baseline that was correlated with the local reactogenicity to vaccination and defined an early signature that correlated with the hemagglutinin antibody titers. These results highlight an adaptive behavior of the innate immune system in evoking a memory-like response to secondary vaccination and define molecular correlates of reactogenicity and immunogenicity in infants.


Comparison of superior-level facet joint violations during open and percutaneous pedicle screw placement.
Superior-level facet joint violation by pedicle screws may result in increased stress to the level above the instrumentation and may contribute to adjacent segment disease. Previous studies have evaluated facet joint violations in open or percutaneous screw cases, but there are no reports describing a direct institutional comparison. To compare the incidence of superior-level facet violation for open vs percutaneous pedicle screws and to evaluate patient and surgical factors that affect this outcome. We reviewed 279 consecutive patients who underwent an index instrumented lumbar fusion from 2007 to 2011 for degenerative spine disease with stenosis with or without spondylolisthesis. We used a computed tomography grading system that represents progressively increasing grades of facet joint violation. Patient and surgical factors were evaluated to determine their impact on facet violation. Our cohort consisted of 126 open and 153 percutaneous cases. Percutaneous procedures had a higher overall violation grade (P = .02) and a greater incidence of high-grade violations (P = .006) compared with open procedures. Bivariate analysis showed significantly greater violations in percutaneous cases for age < 65 years, obesity, pedicle screws at L4, and 1- and 2-level surgeries. Multivariate analysis showed the percutaneous approach and depth of the spine to be independent risk factors for high-grade violations. This study demonstrates greater facet violations for percutaneously placed pedicle screws compared with open screws.


Magnetic properties of the lunar crystalline rock and fines.
Magnetic measurements have shown that nondiamagnetic minerals in a lunar crystalline rock of type B are (free Fe(2)+ in paramagnetic pyroxenes) : (antiferromagnetic FeSiO(3)) : (antiferromagnetic FeTiO(3)) : (ferromagnetic iron) = 4.3 : 7 : 20 : 0.08 in weight percentage. The abundance of ferromagnetic Fe in the lunar fines is about 7.5 times its abundance in the crystalline rock. The natural remanent magnetization of the crystalline rock of 7.5 x 10(-6) emu/ g in intensity may not be attributable to its thermoremanent magnetization.


Rewriting the history of Chinese families in nineteenth-century Australia.
The nineteenth-century Chinese population in Australia was made up mostly of men, drawing many commentators to the conclusion these men faced an absence of family life, resulting in prostitution, gambling, opium use and other so-called vices. Recent research has, however, expanded and complicated our knowledge of Chinese families in New South Wales and Victoria, particularly concerning the extent to which Chinese men and white Australian women formed intimate relationships. This article traces the origins of the misconceptions about Chinese families in nineteenth-century Australia, and considers how new directions in scholarship over the past decade are providing methods for enlarging our knowledge. It argues that instead of being oddities or exceptions, Chinese-European families were integral to the story of Australia's early Chinese communities.


Model-based clustering of multi-tissue gene expression data.
Recently, it has become feasible to generate large-scale, multi-tissue gene expression data, where expression profiles are obtained from multiple tissues or organs sampled from dozens to hundreds of individuals. When traditional clustering methods are applied to this type of data, important information is lost, because they either require all tissues to be analyzed independently, ignoring dependencies and similarities between tissues, or to merge tissues in a single, monolithic dataset, ignoring individual characteristics of tissues. We developed a Bayesian model-based multi-tissue clustering algorithm, revamp, which can incorporate prior information on physiological tissue similarity, and which results in a set of clusters, each consisting of a core set of genes conserved across tissues as well as differential sets of genes specific to one or more subsets of tissues. Using data from seven vascular and metabolic tissues from over 100 individuals in the STockholm Atherosclerosis Gene Expression (STAGE) study, we demonstrate that multi-tissue clusters inferred by revamp are more enriched for tissue-dependent protein-protein interactions compared to alternative approaches. We further demonstrate that revamp results in easily interpretable multi-tissue gene expression associations to key coronary artery disease processes and clinical phenotypes in the STAGE individuals. Revamp is implemented in the Lemon-Tree software, available at https://github.com/eb00/lemon-tree. Supplementary data are available at Bioinformatics online.


Principled Design and Runtime Analysis of Abstract Convex Evolutionary Search.
Geometric crossover is a formal class of crossovers that includes many well-known recombination operators across representations. In previous work, it was shown that all evolutionary algorithms with geometric crossover (but no mutation) do the same form of convex search regardless of the underlying representation, the specific selection mechanism, offspring distribution, search space, and problem at hand. Furthermore, it was suggested that the generalised convex search could perform well on generalised forms of concave and approximately concave fitness landscapes regardless of the underlying space and representation. In this article, we deepen this line of enquiry and study the runtime of generalised convex search on concave fitness landscapes. This is a first step toward linking a geometric theory of representations and runtime analysis in the attempt to (1) set the basis for a more general, unified approach for the runtime analysis of evolutionary algorithms across representations, and (2) identify the essential matching features of evolutionary search behaviour and landscape topography that cause polynomial performance. We present a general runtime result that can be systematically instantiated to specific search spaces and representations and present its specifications to three search spaces. As a corollary, we obtain that the convex search algorithm optimises LeadingOnes in [Formula: see text] fitness evaluations, which is faster than all unbiased unary black box algorithms.


The immobilization of fibronectin- and fibroblast growth factor 2-derived peptides on a culture plate supports the attachment and proliferation of human pluripotent stem cells.
Pluripotent stem cells (PSCs) offer a promising tool for regenerative medicine. The clinical application of PSCs inevitably requires a large-scale culture in a highly defined environment. The present study aimed to devise defined coating materials for the efficient adhesion and proliferation of human PSCs (hPSCs). We tested the activity of seven fibronectin-derived peptides and three laminin-derived peptides for the attachment and proliferation of hPSCs through their immobilization on the bottom of culture dishes by creating a fusion protein with the mussel adhesion protein. Among the extracellular matrix (ECM) mimetics tested, one fibronectin-derived peptide, PHSRN-GRGDSP, significantly promoted adhesion, enhanced alkaline phosphatase activity, and increased pluripotency-related gene expression in hPSCs compared to Matrigel. Furthermore, co-immobilization of a particular canofin peptide derived from fibroblast growth factor 2 increased pluripotency marker expression, which may offer the possibility of culture without growth factor supplementation. Our findings afford a novel defined condition for the efficient culture of hPSCs and may be utilized in future clinical applications.


RD114 envelope proteins provide an effective and versatile approach to pseudotype lentiviral vectors.
Lentiviral vectors derived from the HIV-1 genome offer great promise for gene therapy due to their ability to transduce non-dividing cells and sustain long-term expression of transgenes. The majority of current lentiviral vectors are pseudotyped with the vesicular stomatitis viral envelope (VSV-G). VSV-G equips lentiviral vectors with a broad host cell tropism and increased stability. Increased particle stability enables viral supernatants to be concentrated by high-speed centrifugation to enhance their infectivity. Despite its efficacy, VSV-G is cytotoxic - a feature that prohibits the development of stable cell lines that constitutively express this envelope. Therefore, non-toxic envelope proteins are being investigated. RD114 is an attractive alternative because it also provides increased particle stability and its receptor is widely expressed on hematopoietic stem cells (HSCs). In this study, the packaging efficiency of three envelope proteins, RD114, RDpro and VSV-G, were evaluated with two lentiviral vectors (TRIP GFP and HPV-402). RDpro is an RD114-HIV chimera designed to pseudotype lentiviral vectors more efficiently. In transient systems, VSV-G generated titers of 10(8) and 10(7) viral particles/mL for TRIP GFP and HPV-402. RDpro possessed titers of 10(7) and 10(6), while RD114 titers were one log lower for each vector. Despite having relatively lower titers, RD114 proteins are less toxic; this was demonstrated in the extension of transient transfection reactions from 48 to 96 h. VSV-G transfections are generally limited to 48 h. In regard to gene therapy applications, we show that RDpro supernatants efficiently transduce peripheral blood HSCs. The versatility of RD114 envelopes was again demonstrated by using a 'mixed' expression system; composed of stably expressed RD114 envelope proteins to pseudotype lentiviral vectors generated in trans (titer range 10(3)-10(5)). Our data show that RD114 envelope proteins are effective and versatile constructs that could prove to be essential components of therapeutic lentiviral gene transfer systems.


An NMR and MD study of complexes of bacteriophage lambda lysozyme with tetra- and hexa-N-acetylchitohexaose.
The X-ray structure of lysozyme from bacteriophage lambda (λ lysozyme) in complex with the inhibitor hexa-N-acetylchitohexaose (NAG6) (PDB: 3D3D) has been reported previously showing sugar units from two molecules of NAG6 bound in the active site. One NAG6 is bound with four sugar units in the ABCD sites and the other with two sugar units in the E'F' sites potentially representing the cleavage reaction products; each NAG6 cross links two neighboring λ lysozyme molecules. Here we use NMR and MD simulations to study the interaction of λ lysozyme with the inhibitors NAG4 and NAG6 in solution. This allows us to study the interactions within the complex prior to cleavage of the polysaccharide. 1 HN and 15 N chemical shifts of λ lysozyme resonances were followed during NAG4/NAG6 titrations. The chemical shift changes were similar in the two titrations, consistent with sugars binding to the cleft between the upper and lower domains; the NMR data show no evidence for simultaneous binding of a NAG6 to two λ lysozyme molecules. Six 150 ns MD simulations of λ lysozyme in complex with NAG4 or NAG6 were performed starting from different conformations. The simulations with both NAG4 and NAG6 show stable binding of sugars across the D/E active site providing low energy models for the enzyme-inhibitor complexes. The MD simulations identify different binding subsites for the 5th and 6th sugars consistent with the NMR data. The structural information gained from the NMR experiments and MD simulations have been used to model the enzyme-peptidoglycan complex.


Runx2 trans-activation mediated by the MSX2-interacting nuclear target requires homeodomain interacting protein kinase-3.
Runt-related transcription factor 2 (Runx2) and muscle segment homeobox homolog 2-interacting nuclear target (MINT) (Spen homolog) are transcriptional regulators critical for mammalian development. MINT enhances Runx2 activation of osteocalcin (OC) fibroblast growth factor (FGF) response element in an FGF2-dependent fashion in C3H10T1/2 cells. Although the MINT N-terminal RNA recognition motif domain contributes, the muscle segment homeobox homolog 2-interacting domain is sufficient for Runx2 activation. Intriguingly, Runx1 cannot replace Runx2 in this assay. To better understand this Runx2 signaling cascade, we performed structure-function analysis of the Runx2-MINT trans-activation relationship. Systematic truncation and domain swapping in Runx1:Runx2 chimeras identified that the unique Runx2 activation domain 3 (AD3), encompassed by residues 316-421, conveys MINT+FGF2 trans-activation in transfection assays. Ala mutagenesis of Runx2 Ser/Thr residues identified that S301 and T326 in AD3 are necessary for full MINT+FGF2 trans-activation. Conversely, phosphomimetic Asp substitution of these AD3 Ser/Thr residues enhanced activation by MINT. Adjacent Pro residues implicated regulation by a proline-directed protein kinase (PDPK). Systematic screening with PDPK inhibitors identified that the casein kinase-2/homeodomain-interacting protein kinase (HIPK)/dual specificity tyrosine phosphorylation regulated kinase inhibitor 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT), but not ERK, c-Jun N-terminal kinase, p38MAPK, or other casein kinase-2 inhibitors, abrogated Runx2-, MINT-, and FGF2-activation. Systematic small interfering RNA-mediated silencing of DMAT-inhibited PDPKs revealed that HIPK3 depletion reduced MINT+FGF2-dependent activation of Runx2. HIPK3 and Runx2 coprecipitate after in vitro transcription-translation, and recombinant HIPK3 recognizes Runx2 AD3 as kinase substrate. Furthermore, DMAT treatment and HIPK3 RNAi inhibited MINT+FGF2 activation of Runx2 AD3, and nuclear HIPK3 colocalized with MINT. HIPK3 antisense oligodeoxynucleotide selectively reduced Runx2 protein accumulation and OC gene expression in C3H10T1/2 cells. Thus, HIPK3 participates in MINT+FGF2 regulation of Runx2 AD3 activity and controls Runx2-dependent OC expression.


Fibulin-3 as a diagnostic biomarker in patients with malignant mesothelioma.
New tumour biomarkers are being intensely investigated for malignant mesothelioma (MM). Fibulin-3 is produced in MM but its role remains uncertain. The aim of this study was to evaluate the validity of measuring serum fibulin-3 in the diagnosis and prognosis of MM. This prospective study was performed on 43 patients and 40 healthy controls who were admitted to our hospital between January 2012 and January 2014. Data from MM patients, including demographic and clinical features, routine laboratory data, levels of serum fibulin-3, and treatment outcomes were defined as potential prognostic factors. The receiver operating characteristic (ROC) curve for fibulin-3 was used to detect the cut-off value with highest sensitivity and specificity. Univariate survival analysis was performed using the Kaplan-Meier method in patients with MM. Afterwards, the possible factors identified with univariate analyses were entered into the cox regression analysis. Our results revealed that patients with MM had significantly higher serum levels of fibulin-3 than controls. The results showed that the best cut-off point was 36.6 ng/ml with an AUC (area under the curve)=0.976, sensitivity=93.0% and specificity=90.0. In our study, the initial significant poor prognostic factors were advanced stage, high white blood cell count, high platelet count, high C-reactive protein (p<0.05 for each variable). Later, according to multivariate analysis the results showed only advanced stage as significant parameter (p=0.040). We determined that real use for serum fibulin-3 was not for prognosis but for diagnosis in MM. Also advanced stage was associated with poor MM prognosis.