Randall Selitto pressure algometry for assessment of bone-related pain in rats.
Deep pain is neglected compared with cutaneous sources. Pressure algometry has been validated in the clinic for assessment of bone-related pain in humans. In animal models of bone-related pain, we have validated the Randall Selitto behavioural test for assessment of acute and pathological bone pain and compared the outcome with more traditional pain-related behaviour measures. Randall Selitto pressure algometry was performed over the anteromedial part of the tibia in naïve rats, sham-operated rats, and rats inoculated with MRMT-1 carcinoma cells in the left tibia, and the effect of morphine was investigated. Randall Selitto measures of cancer-induced bone pain were supplemented by von Frey testing, weight-bearing and limb use test. Contribution of cutaneous nociception to Randall Selitto measures were examined by local anaesthesia. Randall Selitto pressure algometry over the tibia resulted in reproducible withdrawal thresholds, which were dose-dependently increased by morphine. Cutaneous nociception did not contribute to Randall Selitto measures. In cancer-bearing animals, compared with sham, significant differences in pain-related behaviours were demonstrated by the Randall Selitto test on day 17 and 21 post-surgery. A difference was also demonstrated by von Frey testing, weight-bearing and limb use tests. Our results indicate that pressure applied by the Randall Selitto algometer on a region, where the bone is close to the skin, may offer a way to measure bone-related pain in animal models and could provide a supplement to the traditional behavioural tests and a means to study deep pain.


Longitudinal Dynamics of a Blood Transcriptomic Signature of Tuberculosis.
Rationale: Performance of blood transcriptomic tuberculosis (TB) signatures in longitudinal studies and effects of TB-preventive therapy and coinfection with HIV or respiratory organisms on transcriptomic signatures has not been systematically studied. Objectives: We evaluated longitudinal kinetics of an 11-gene blood transcriptomic TB signature, RISK11, and effects of TB-preventive therapy (TPT) and respiratory organisms on RISK11 signature score, in HIV-uninfected and HIV-infected individuals. Methods: RISK11 was measured in a longitudinal study of RISK11-guided TPT in HIV-uninfected adults, a cross-sectional respiratory organisms cohort, or a longitudinal study in people living with HIV (PLHIV). HIV-uninfected RISK11+ participants were randomized to TPT or no TPT; RISK11- participants received no TPT. PLHIV received standard-of-care antiretroviral therapy and TPT. In the cross-sectional respiratory organisms cohort, viruses and bacteria in nasopharyngeal and oropharyngeal swabs were quantified by real-time quantitative PCR. Measurements and Main Results: RISK11+ status was transient in most of the 128 HIV-negative participants with longitudinal samples; more than 70% of RISK11+ participants reverted to RISK11- by 3 months, irrespective of TPT. By comparison, reversion from a RISK11+ state was less common in 645 PLHIV (42.1%). Non-HIV viral and nontuberculous bacterial organisms were detected in 7.2% and 38.9% of the 1,000 respiratory organisms cohort participants, respectively, and among those investigated for TB, 3.8% had prevalent disease. Median RISK11 scores (%) were higher in participants with viral organisms alone (46.7%), viral and bacterial organisms (42.8%), or prevalent TB (85.7%) than those with bacterial organisms other than TB (13.4%) or no organisms (14.2%). RISK11 could not discriminate between prevalent TB and viral organisms. Conclusions: Positive RISK11 signature status is often transient, possibly due to intercurrent viral infection, highlighting potentially important challenges for implementation of these biomarkers as new tools for TB control.


Breast Milk, a Source of Beneficial Microbes and Associated Benefits for Infant Health.
Human breast milk is considered the optimum feeding regime for newborn infants due to its ability to provide complete nutrition and many bioactive health factors. Breast feeding is associated with improved infant health and immune development, less incidences of gastrointestinal disease and lower mortality rates than formula fed infants. As well as providing fundamental nutrients to the growing infant, breast milk is a source of commensal bacteria which further enhance infant health by preventing pathogen adhesion and promoting gut colonisation of beneficial microbes. While breast milk was initially considered a sterile fluid and microbes isolated were considered contaminants, it is now widely accepted that breast milk is home to its own unique microbiome. The origins of bacteria in breast milk have been subject to much debate, however, the possibility of an entero-mammary pathway allowing for transfer of microbes from maternal gut to the mammary gland is one potential pathway. Human milk derived strains can be regarded as potential probiotics; therefore, many studies have focused on isolating strains from milk for subsequent use in infant health and nutrition markets. This review aims to discuss mammary gland development in preparation for lactation as well as explore the microbial composition and origins of the human milk microbiota with a focus on probiotic development.


Gastric cancer and Helicobacter pylori infection.
To identify differences in the prevalence of Helicobacter pylori infection in different groups of patients with gastric cancer. In total 224 cases of gastric cancer were studied: 120 (53.6%) intestinal; 69 (30.8%) diffuse; and 35 (15.6%) unclassified. Site of tumour, presence and severity of gastritis, presence and extent of intestinal metaplasia, and age and sex were also recorded. Infection by H pylori was assessed using modified Giemsa staining. H pylori infection was found in 96 (43%) cases. There was no significant association between infection and histological type of tumour, nor was there any significant association between infection and site of tumour, the presence of intestinal metaplasia, age, or sex. The only significant association identified was between infection and gastritis. These results are in broad agreement with those of other similar studies, although the overall prevalence of infection, at 43%, was lower than has been reported in some series. The findings do not support a role for H pylori in any particular subgroup of patients with gastric cancer but do not exclude a role for the organism in the pathogenesis of gastric cancer as a whole.


Effects of New World scorpion toxins on single-channel and whole cell cardiac sodium currents.
Purified toxins from a North American scorpion, Centruroides noxius (Cn II-10), and a South American scorpion, Tityus serrulatus (Ts-gamma), were tested on cardiac sodium channels using patch-clamp methods to record whole cell and single-channel currents. The two toxins produced similar effects on sodium currents; potassium and calcium currents were not affected. Macroscopic sodium current amplitudes, measured at test potentials greater than -20 mV where the opening probability was high, decreased in a concentration-dependent manner with a half maximum inhibitory concentration of 6 X 10(-8) M. Block was unchanged by repetitive depolarizing pulses. In the presence of scorpion toxin, the currents were rapidly blocked by tetrodotoxin (3 X 10(-5) M). Both toxins shifted the voltage dependence of sodium channel inactivation to more negative potentials. At test potentials between -50 and -70 mV, where the sodium channel opening probability is normally low, both toxins produced an increase in sodium current and slowed the rates of activation and inactivation. At intermediate potentials between -50 and -20 mV the currents in the presence of toxins crossed over the control currents. At a test potential of -20 mV, the toxins decreased single-channel activity and increased the latency to first opening. At a test potential of -60 mV, the toxins significantly prolonged channel open time. The unitary current amplitudes were unchanged at either potential. We conclude that New World scorpion toxins produce apparently complex effects on whole cell currents primarily by retarding activation gating of cardiac sodium channels.


Transcription of the Escherichia coli fatty acid synthesis operon fabHDG is directly activated by FadR and inhibited by ppGpp.
In Escherichia coli, FadR and FabR are transcriptional regulators that control the expression of fatty acid degradation and unsaturated fatty acid synthesis genes, depending on the availability of fatty acids. In this report, we focus on the dual transcriptional regulator FadR. In the absence of fatty acids, FadR represses the transcription of fad genes required for fatty acid degradation. However, FadR is also an activator, stimulating transcription of the products of the fabA and fabB genes responsible for unsaturated fatty acid synthesis. In this study, we show that FadR directly activates another fatty acid synthesis promoter, PfabH, which transcribes the fabHDG operon, indicating that FadR is a global regulator of both fatty acid degradation and fatty acid synthesis. We also demonstrate that ppGpp and its cofactor DksA, known primarily for their role in regulation of the synthesis of the translational machinery, directly inhibit transcription from the fabH promoter. ppGpp also inhibits the fadR promoter, thereby reducing transcription activation of fabH by FadR indirectly. Our study shows that both ppGpp and FadR have direct roles in the control of fatty acid promoters, linking expression in response to both translation activity and fatty acid availability.


Quantitative kinetics of the atmospheric reaction between isocyanic acid and hydroxyl radicals: post-CCSD(T) contribution, anharmonicity, recrossing effects, torsional anharmonicity, and tunneling.
Hydroxyl radicals (OH) are the most important atmospheric oxidant, initiating atmospheric reactions for the chemical transformation of volatile organic compounds. Here, we choose the HNCO + OH reaction as a prototype reaction because it contains the fundamental reaction processes for OH radicals: H-abstraction reaction by OH and OH addition reaction. However, its kinetics are unknown under atmospheric conditions. We investigate the reaction of HNCO with OH by using the GMM(P).L method close to the accuracy of single, double, triple, and quadruple excitations and noniterative quintuple excitations with a complete basis set (CCSDTQ(P)/CBS) as benchmark results and a dual-level strategy for kinetics calculations. The calculated rate constant of HNCO + OH is in good agreement with the experimental data available at the temperatures between 620 and 2500 K. We find that the rate constant cannot be correctly obtained by using experimental data to extrapolate the atmospheric temperature ranges. We find that the post-CCSD(T) contribution is very large for the barrier height with the value of -0.85 kcal mol-1 for the H-abstraction reaction, while the previous investigations were done up to the CCSD(T) level. Moreover, we also find that recrossing effects, tunneling, torsional anharmonicity, and anharmonicity are important for obtaining quantitative kinetics in the OH + HNCO reaction.


The potential for a selective screening strategy for abdominal aortic aneurysm.
To investigate the feasibility of selective screening for abdominal aortic aneurysm (AAA) based on identification of a target group of manageable size defined by risk factors for AAA. Male residents of Perth, Western Australia, aged 65-83 years, who participated in a randomised controlled trial of ultrasound screening for AAA. Eligible men were identified from the electoral roll and invited to attend a screening clinic. Those who attended completed a questionnaire, had a limited physical examination, and underwent an ultrasound examination to identify the maximum diameter of the infrarenal aorta. Data on risk factors collected from the first 8995 men seen were used to calculate a multivariate risk score for the remaining 2755 men who were screened. Centiles of the risk score were used to define potential target groups for screening and the sensitivity and specificity of each of these selective screening strategies were calculated. We repeated the calculation separately for AAAs of at least 30 mm, 40 mm, and 50 mm in diameter. We found that screening half of the male population aged 65-83 years would find approximately 75% of AAAs, regardless of their size, whereas screening only current smokers in this population would find approximately 20% of AAAs. Selective screening for AAA using easily recognisable risk factors is feasible but is not worthwhile as approximately 25% ofclinically significant cases would be missed.


Mortality impact of AIDS in Addis Ababa, Ethiopia.
To measure the impact of HIV on mortality in Addis Ababa, Ethiopia. A retrospective review of burials at three cemeteries, 1987-2001 and a prospective surveillance of burials at all (n = 70) cemeteries, February-May, 2001. The age, sex, and date of burial were recorded; in the absence of denominators, we compared the ratio of deaths of persons 25-49 versus 5-14 years of age per calendar year, using logistic regression, adjusting for sex and site. The age- and sex-specific mortality were calculated and compared with pre-HIV mortality in 1984. Of 17,519 deaths, retrospectively reviewed, complete data were available for 6342 (47%) females and 7269 (53%) males. During 1987-2001, the '25-49' versus '5-14' group all-cause mortality ratio increased by 8.5% per calendar year (P < 0.05). A total of 5101 deaths were recorded in the prospective surveillance. Crude mortality rates were 9.5/1000 per year (men) and 7.1/1000 per year (women). In comparison with 1984, 5.0-times as many men and 5.3-times as many women died in the age group 35-39 years. Attributing the increase in mortality in ages 15-60 to HIV in the period 1984-2001, Ethiopian men and women have a probability of 18.8 and 17.8%, respectively, of dying of HIV before age 60. Burials increased significantly among the '25-49', versus the '5-14' group, during the period 1987-2001. This trend, and a five-times higher mortality in 2001 than in 1984 in those aged 35-39 years demonstrate a severe impact of HIV on mortality. Continuing surveillance of burials is recommended.


A task taxonomy for network evolution analysis.
Visualization has proven to be a useful tool for understanding network structures. Yet the dynamic nature of social media networks requires powerful visualization techniques that go beyond static network diagrams. To provide strong temporal network visualization tools, designers need to understand what tasks the users have to accomplish. This paper describes a taxonomy of temporal network visualization tasks. We identify the 1) entities, 2) properties, and 3) temporal features, which were extracted by surveying 53 existing temporal network visualization systems. By building and examining the task taxonomy, we report which tasks are well covered by existing systems and make suggestions for designing future visualization tools. The feedback from 12 network analysts helped refine the taxonomy.


Importance of critical micellar concentration for the prediction of solubility enhancement in biorelevant media.
This study evaluated if the intrinsic surface properties of compounds are related to the solubility enhancement (SE) typically observed in biorelevant media like fasted state simulated intestinal fluids (FaSSIF). The solubility of 51 chemically diverse compounds was measured in FaSSIF and in phosphate buffer and the surface activity parameters were determined. This study showed that the compound critical micellar concentration parameter (CMC) correlates strongly with the solubility enhancement (SE) observed in FaSSIF compared to phosphate buffer. Thus, the intrinsic capacity of molecules to form micelles is also a determinant for each compound's affinity to the micelles of biorelevant surfactants. CMC correlated better with SE than lipophilicity (logD), especially over the logD range typically covered by drugs (2 < logD < 4). CMC can become useful to guide drug discovery scientists to better diagnose, improve, and predict solubility in biorelevant media, thereby enhancing oral bioavailability of drug candidates.


Polymorphous ventricular tachycardia associated with acute myocardial infarction.
During a 2.9-year period, 11 patients developed polymorphous ventricular tachycardia 1-13 days after acute anterior (seven patients) or inferior (four patients) myocardial infarction. None of the 11 patients had sinus bradycardia (mean heart rate, 90 +/- 23 beats/min), but three had a sinus pause immediately before the onset of polymorphous ventricular tachycardia. In all 11 patients, the QT interval and corrected QT interval (QTc) were normal or minimally prolonged (QT, 385 +/- 34 msec; QTc, 442 +/- 40 msec). None had significant hypokalemia (mean serum potassium concentration, 4.3 +/- 0.5 meq/l) or a grossly abnormal serum magnesium or calcium concentration (2.1 +/- 0.4 and 8.9 +/- 0.7 mg/dl, respectively). Immediately before the onset of polymorphous ventricular tachycardia, symptoms and/or electrocardiographic changes consistent with recurrent myocardial ischemia occurred in nine of 11 patients. One patient died before drug therapy could be initiated. Lidocaine was used in 10 patients and proved to be effective in only one. Intravenous procainamide was used in six patients: one improved, and five had recurrence of polymorphous ventricular tachycardia. Bretylium was used in five patients and was ineffective in all cases. Overdrive pacing was used in four patients and failed to suppress recurrent arrhythmias in all cases. Four patients with persistent polymorphous ventricular tachycardia unresponsive to lidocaine, procainamide, or bretylium responded to intravenous amiodarone. One patient with polymorphous ventricular tachycardia that was consistently preceded by ST segment elevation responded to intravenous nitroglycerin. Two patients with persistent polymorphous ventricular tachycardia and obvious recurrent ischemia unresponsive to pharmacological intervention responded to emergency coronary revascularization. A third patient who experienced recurrent angina and polymorphous tachycardia was initially stabilized with pharmacological therapy but subsequently underwent elective revascularization and has remained stable without antiarrhythmic therapy. Post-myocardial infarction polymorphous ventricular tachycardia is not consistently related to an abnormally long QT interval, sinus bradycardia, preceding sinus pauses, or electrolyte abnormalities. This arrhythmia has a variable response to class I antiarrhythmics but may be suppressed by intravenous amiodarone therapy. It is often associated with signs or symptoms of recurrent myocardial ischemia. Furthermore, coronary revascularization appears to be effective in preventing the recurrence of polymorphous ventricular tachycardia when associated with recurrent postinfarction angina.


Molecular and Cellular Interactions of Scavenger Receptor SR-F1 With Complement C1q Provide Insights Into Its Role in the Clearance of Apoptotic Cells.
The scavenger receptor SR-F1 binds to and mediates the internalization of a wide range of ligands, and is involved in several immunological processes. We produced recombinant SR-F1 ectodomain and fragments deleted from the last 2 or 5 C-terminal epidermal growth factor-like modules and investigated their role in the binding of acetylated low density lipoprotein (AcLDL), complement C1q, and calreticulin (CRT). C1q measured affinity was in the 100 nM range and C1q interaction occurs via its collagen-like region. We identified two different binding regions on SR-F1: the N-terminal moiety interacts with C1q and CRT whereas the C-terminal moiety binds AcLDL. The role of SR-F1 N-linked glycans was also tested by mutating each of the three glycosylated asparagines. The three mutants retained binding activities for both AcLDL and C1q. A stable THP-1 cell line overexpressing SR-F1 was generated and C1q was shown to bind more strongly to the surface of SR-F1 overexpressing macrophages, with C1q/SR-F1 colocalization observed in some membrane areas. We also observed a higher level of CRT internalization for THP-1 SR-F1 cells. Increasing SR-F1 negatively modulated the uptake of apoptotic cells. Indeed, THP-1 cells overexpressing SR-F1 displayed a lower phagocytic capacity as compared with mock-transfected cells, which could be partially restored by addition of C1q in the extracellular milieu. Our data shed some light on the role of SR-F1 in efferocytosis, through its capacity to bind C1q and CRT, two proteins involved in this process.


The value of Ret-Hb and sTfR in the diagnosis of iron depletion in healthy, young children.
Reticulocyte hemoglobin (Ret-Hb) content and soluble transferrin receptor (sTfR) are described as promising biomarkers in the analysis of iron status. However, the value of Ret-Hb and sTfR in the early detection of iron depletion, as frequently observed in children in high-income countries, is unclear. We hypothesized that young children to iron depletion, using the WHO cutoff of ferritin <12 μg/l, would have lower Ret-Hb and higher sTfR concentrations compared to children with a ferritin ⩾level 12 μg/l. In this cross-sectional study, we analyzed mean concentrations of Ret-Hb and sTfR in 351 healthy children aged 0.5-3 years in a high-income country. The Student's t-test was used to compare Ret-Hb and sTfR concentrations between groups. We showed that concentrations of Ret-Hb and sTfR are similar in children with and without iron depletion. A decrease in Ret-Hb concentration was present only when ferritin concentrations were <8 μg/l. sTfR concentrations were similar in children with ferritin concentrations <6 μg/l and ⩾12 μg/l. Our results showed that the discriminative value of Ret-Hb and sTfR for the detection of iron depletion is limited. Our findings suggest that ferritin is the most useful biomarker in the screening of iron depletion in healthy children in high-income countries. However, ideally, reference ranges of iron status biomarkers should be based on studies showing that children with concentrations outside reference ranges have poor neurodevelopmental outcomes.


Male-female differences in households' resource allocation and decision to seek healthcare in south-eastern Nigeria: Results from a mixed methods study.
Ability to influence household decision-making has been shown to increase with improved social capital and power and is linked to better access to household financial resources and other services outside the household including healthcare. To examine the male-female differences in household custody of financial resources, decision-making, and type of healthcare utilised, we used a mixed methods approach of cross-sectional household surveys and focus-group discussions (FGDs). Data was collected between 10 January-28 February 2011. We analyzed a sample of 411 households and a sub-sample of 223 households with a currently married head. We conducted six single-sex FGDs in 3 communities (1 urban, 2 rural) among a random sub-sample of participants in the survey. We performed univariate, bivariate, and logistic regression analyses with a 95% confidence interval. For the qualitative data, we performed thematic analysis where broad themes relevant to the research objective were abstracted. In all households and in those with a married head, sick male members were less likely to forgo healthcare (aORall0.87, 95% CI 0.80-0.90; aORmarried0.52, 95% CI 0.18-0.83) and more likely to utilise formal healthcare relative to female sick members (aORall3.36, 95% CI 3.20-3.87; aORmarried19.50, 95% CI 9.62-39.52). Formal healthcare providers are medically trained while informal providers are untrained vendors that dispense medications for profit. There were more reports of sole custody of household resources among men within households with married heads. Joint decision-making on healthcare expenditure improved women's access to healthcare but is not reflective of unhindered access to household financial resources. Qualitatively, women spoke of seeking permission from male household head before expenditure was incurred, while male heads spoke of concealing household financial resources from their spouse. Gender constructs and male-female differences have important effects on household resource allocation and healthcare utilisation.


Temporal-Spatial Transcriptome Analyses Provide Insights into the Development of Petaloid Androecium in Canna indica.
Canna indica (Zingiberales) is one of the most important ornamental species characterized with beautiful petaloid staminodes, which are considered to evolve from stamens. However, the genetic basis for the development of petaloid staminodes remains unclear largely because the genomic sequences are not available. By using RNA-Seq, we sequenced the transcripts in the flower of C. indica, and quantified the temporal gene expressions in flower primordium and differentiated flower, as well as the spatial gene expressions in petal and petaloid staminode. In total, 118,869 unigenes were assembled, among which 67,299 unigenes were annotated. Quantification analysis identified the differentially expressed genes in the temporal and spatial two comparisons, based on which, Gene Ontology enrichment analysis highlighted the representative terms in each sample, such as specification of organ number in flower primordium, growth in differentiated flower, secondary cell wall biogenesis in petal and cell division in petaloid staminode. Among the 51 analyzed MADS-box unigenes, 37 were up-regulated in differentiated flower compared with those in flower primordium. A-class unigenes were expressed higher in petal than in petaloid staminode, and C-class unigenes were expressed oppositely, whereas B-class unigenes demonstrated close expression levels in these two organs, indicating that petaloid staminode retains stamen identity to some degree. In situ hybridization provided more detailed expression patterns of these unigenes, and revealed the extended expression of B-class to the carpel at later stages when the style turned flat. These results constitute a preliminary basis for the study of flower development in C. indica and can be applied in further study of the evolution of Zingiberales.


Distinctive characteristics of sexual orientation bias crimes.
Despite increased attention in the area of hate crime research in the past 20 years, sexual orientation bias crimes have rarely been singled out for study. When these types of crimes are looked at, the studies are typically descriptive in nature. This article seeks to increase our knowledge of sexual orientation bias by answering the question: What are the differences between sexual orientation motivated bias crimes and racial bias crimes? This question is examined using data from the National Incident Based Reporting System (NIBRS) and multiple regression techniques. This analysis draws on the strengths of NIBRS to look at the incident characteristics of hate crimes and distinguishing characteristics of sexual orientation crimes. Specifically this analysis looks at the types and seriousness of offenses motivated by sexual orientation bias as opposed to race bias as well as victim and offender characteristics. The findings suggest that there are differences between these two types of bias crimes, suggesting a need for further separation of the bias types in policy and research.


A patient with typical clinical features of mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS) but without an obvious genetic cause: a case report.
There are currently 23 missense point mutations and one 4 basepair deletion spanning different mitochondrial genes associated with mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS). The spectrum of mitochondrial DNA mutations in Arab patients with MELAS is largely unknown. A standard clinical examination was carried out on a 34-year-old Saudi woman showing clinical features of MELAS. Fresh frozen muscle tissue was subjected to enzyme histochemical analysis. DNA was extracted from her leukocytes and muscle tissue, and the full mitochondrial genome was screened for base substitution mutations and deletions. Additionally, we screened the polymerase gamma-1 nuclear gene for mutations. The patient was negative for the most common m.3243 A>G MELAS mutation. Sequencing the full mitochondrial genome did not reveal any known or potentially pathogenic sequence changes. The polymerase gamma-1 gene was also free from mutations. The clinical picture described here typically fits that observed in patients with MELAS or mitochondrial stroke-like events, but mutations in recognized genes (mitochondrial DNA and polymerase gamma-1 gene) were absent. We report the case of a patient with typical clinical features of MELAS, but without an obvious genetic cause.


Tracking TCRβ Sequence Clonotype Expansions during Antiviral Therapy Using High-Throughput Sequencing of the Hypervariable Region.
To maintain a persistent infection viruses such as hepatitis C virus (HCV) employ a range of mechanisms that subvert protective T cell responses. The suppression of antigen-specific T cell responses by HCV hinders efforts to profile T cell responses during chronic infection and antiviral therapy. Conventional methods of detecting antigen-specific T cells utilize either antigen stimulation (e.g., ELISpot, proliferation assays, cytokine production) or antigen-loaded tetramer staining. This limits the ability to profile T cell responses during chronic infection due to suppressed effector function and the requirement for prior knowledge of antigenic viral peptide sequences. Recently, high-throughput sequencing (HTS) technologies have been developed for the analysis of T cell repertoires. In the present study, we have assessed the feasibility of HTS of the TCRβ complementarity determining region (CDR)3 to track T cell expansions in an antigen-independent manner. Using sequential blood samples from HCV-infected individuals undergoing antiviral therapy, we were able to measure the population frequencies of >35,000 TCRβ sequence clonotypes in each individual over the course of 12 weeks. TRBV/TRBJ gene segment usage varied markedly between individuals but remained relatively constant within individuals across the course of therapy. Despite this stable TRBV/TRBJ gene segment usage, a number of TCRβ sequence clonotypes showed dramatic changes in read frequency. These changes could not be linked to therapy outcomes in the present study; however, the TCRβ CDR3 sequences with the largest fold changes did include sequences with identical TRBV/TRBJ gene segment usage and high junction region homology to previously published CDR3 sequences from HCV-specific T cells targeting the HLA-B*0801-restricted (1395)HSKKKCDEL(1403) and HLA-A*0101-restricted (1435)ATDALMTGY(1443) epitopes. The pipeline developed in this proof of concept study provides a platform for the design of future experiments to accurately address the question of whether T cell responses contribute to SVR upon antiviral therapy. This pipeline represents a novel technique to analyze T cell dynamics in situations where conventional antigen-dependent methods are limited due to suppression of T cell functions and highly diverse antigenic sequences.


Effects of sodium vanadate on various types of vascular smooth muscles.
Effects of sodium vanadate on various vascular smooth muscles of guinea pigs, rabbits, and Wistar Kyoto rats (WKY) were studied. Sodium vanadate of concentrations higher than 10(-5) M induced contractions in the aortae of all animals. The contractile effects varied among vascular smooth muscles, and mesenteric arteries showed no or only weak contractile response to the drug, while aortae showed higher contractile responses. In the portal veins, potentiation of spontaneous contractions was observed by the application of sodium vanadate. These responses were not blocked by treatments with adrenergic blocking agents or indomethacin, indicating the direct action of the drug on vascular smooth muscles. Treatment with 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) blocked completely the contractile effects of sodium vanadate, whereas it showed no effect on K-contractures. Partial depolarization of the membrane by elevations of K concentration potentiated sodium vanadate-induced contractions and minimized the variations of responses among preparations. In K-depolarized preparations, sodium vanadate often induced relaxation of preparations. The contractile effects of sodium vanadate were not blocked by treatment with ouabain, though ouabain also showed contractile actions in a number of preparations. It was suggested that vanadate acts directly on vascular smooth muscles and causes contractions without relation to the inhibition of Na,K-ATPase. It may cause contractions inhibiting Ca-ATPase of sarcoplasmic reticulum and/or of cell membrane, and cause relaxation by inhibiting ATPase of contractile proteins. The variations of the responses may be explained by differences of membrane permeability to vanadate.


Results of surgery in idiopathic faecal incontinence.
The surgical treatment of idiopathic faecal incontinence remains a difficult problem, particularly in cases where advanced neuropathy is present. The physiological basis for the post-anal repair has now been questioned and the long-term functional results seem to be rather unsatisfactory. Pelvic floor repair is a more comprehensive approach with encouraging initial results, which have now been shown in a prospective randomised trial to be superior to those of conventional post-anal repair. Gracilis transposition in idiopathic faecal incontinence has shown very disappointing results and probably has no role in the treatment of this condition. In very advanced cases intestinal stoma may occasionally provide important hygienic and psychological benefits.


Embedding an enriched environment in an acute stroke unit increases activity in people with stroke: a controlled before-after pilot study.
To determine whether an enriched environment embedded in an acute stroke unit could increase activity levels in acute stroke patients and reduce adverse events. Controlled before-after pilot study. An acute stroke unit in a regional Australian hospital. Acute stroke patients admitted during (a) initial usual care control period, (b) an enriched environment period and (c) a sustainability period. Usual care participants received usual one-on-one allied health intervention and nursing care. The enriched environment participants were provided stimulating resources, communal areas for eating and socializing and daily group activities. Change management strategies were used to implement an enriched environment within existing staffing levels. Behavioural mapping was used to estimate patient activity levels across groups. Participants were observed every 10 minutes between 7.30 am and 7.30 pm within the first 10 days after stroke. Adverse and serious adverse events were recorded using a clinical registry. The enriched environment group ( n = 30, mean age 76.7 ± 12.1) spent a significantly higher proportion of their day engaged in 'any' activity (71% vs. 58%, P = 0.005) compared to the usual care group ( n = 30, mean age 76.0 ± 12.8). They were more active in physical (33% vs. 22%, P < 0.001), social (40% vs. 29%, P = 0.007) and cognitive domains (59% vs. 45%, P = 0.002) and changes were sustained six months post implementation. The enriched group experienced significantly fewer adverse events (0.4 ± 0.7 vs.1.3 ± 1.6, P = 0.001), with no differences found in serious adverse events (0.5 ± 1.6 vs.1.0 ± 2.0, P = 0.309). Embedding an enriched environment in an acute stroke unit increased activity in stroke patients.


Coordination chemistry of N-heterocyclic nitrenium-based ligands.
Comprehensive studies on the coordination properties of tridentate nitrenium-based ligands are presented. N-heterocyclic nitrenium ions demonstrate general and versatile binding abilities to various transition metals, as exemplified by the synthesis and characterization of Rh(I) , Rh(III) , Mo(0) , Ru(0) , Ru(II) , Pd(II) , Pt(II) , Pt(IV) , and Ag(I) complexes based on these unusual ligands. Formation of nitrenium-metal bonds is unambiguously confirmed both in solution by selective (15) N-labeling experiments and in the solid state by X-ray crystallography. The generality of N-heterocyclic nitrenium as a ligand is also validated by a systematic DFT study of its affinity towards all second-row transition and post-transition metals (Y-Cd) in terms of the corresponding bond-dissociation energies.


European guidelines on perioperative venous thromboembolism prophylaxis: Day surgery and fast-track surgery.
: In recent years, day surgery and fast-track surgery have experienced a continuous increase in volume. Many procedures are now performed on an outpatient protocol, including general, orthopaedic, oncological, reconstructive or vascular surgery. The management of these patients is safe, but the incidence of venous thromboembolism in this population remains unknown. Several risk factors can be identified and stratified derived from studies of inpatient surgical management (e.g. Caprini score). Recommendations for thromboprophylaxis should be tailored from the assessment of both personal and procedure-related risk factors, although with a lack of evidence for application in outpatient management. For patients undergoing a low-risk procedure without additional risk factors, we recommend only general measures of thromboprophylaxis (early ambulation, optimal hydration) (Grade 1B). For patients undergoing a low-risk procedure with additional risk factors, or a high-risk procedure without additional risk factors, we recommend general measures of thromboprophylaxis (Grade 1B) and we suggest the administration of pharmacological prophylaxis with low molecular weight heparins (Grade 2B). For patients undergoing a high-risk procedure with additional risk factors we recommend general measures of thromboprophylaxis (Grade 1B) and pharmacological prophylaxis with low molecular weight heparins over other drugs (Grade 1B), or suggest specific mechanical measures in case of increased bleeding risk (Grade 2C). Pharmacological prophylaxis should last a minimum of 7 days (Grade 1B), although in selected cases of fast-track surgery, thromboprophylaxis could be limited to hospitalisation only (Grade 2C) and in specific cases of high-risk procedures, thromboprophylaxis could be extended for up to 4 weeks (Grade 2B).


Expression and purification of single cysteine-containing mutant variants of the mouse prion protein by oxidative refolding.
The folding and aggregation of proteins has been studied extensively, using multiple probes. To facilitate such experiments, introduction of spectroscopically-active moieties in to the protein of interest is often necessary. This is commonly achieved by specifically labelling cysteine residues in the protein, which are either present naturally or introduced artificially by site-directed mutagenesis. In the case of the recombinant prion protein, which is normally expressed in inclusion bodies, the presence of the native disulfide bond complicates the correct refolding of single cysteine-containing mutant variants of the protein. To overcome this major bottleneck, a simple purification strategy for single tryptophan, single cysteine-containing mutant variants of the mouse prion protein is presented, with yields comparable to that of the wild type protein. The protein(s) obtained by this method are correctly folded, with a single reduced cysteine, and the native disulfide bond between residues C178 and C213 intact. The β-sheet rich oligomers formed from these mutant variant protein(s) are identical to the wild type protein oligomer. The current strategy facilitates sample preparation for a number of high resolution spectroscopic measurements for the prion protein, which specifically require thiol labelling.


Breakdown of mucosal immunity in the gut and resultant systemic sensitization by oral antigens in a murine model for systemic lupus erythematosus.
Secreted IgA plays a pivotal role in the mucosal immunity to maintain the front line of body defense. We found that the level of fecal IgA was dramatically decreased in aged (NZB x NZW)F(1) (BWF(1)) mice developing lupus nephritis, whereas levels in similarly aged New Zealand Black (NZB) and New Zealand White (NZW) mice remained unchanged compared with young mice. The number of cells obtained from Peyer's patches was markedly decreased in aged BWF(1) mice. Aged BWF(1) mice showed increased susceptibility to pathogenic bacterial infection. Furthermore, oral administration of OVA failed to inhibit secondary IgG response induced by systemic immunization, suggesting defective oral tolerance in aged BWF(1) mice. A significant amount of orally administered OVA was incorporated directly into the intestinal lamina propria in aged BWF(1) mice whereas it was mainly localized in subepithelial domes and interfollicular region in Peyer's patches in young mice. T cells obtained from renal and pulmonary lymph nodes of aged BWF(1) mice that had been orally administered with OVA showed an Ag-specific T cell proliferation, whereas those from young BWF(1), aged NZB, and aged NZW mice did not. Interestingly, aerosol exposure to OVA of aged BWF(1) mice, which had been orally administered with the same Ag, provoked an eosinophil infiltration in the lung. These results demonstrate that mucosal immunity in the gut is impaired and oral Ags induce systemic sensitization instead of oral tolerance in the development of murine lupus.


[Liver transplantation from elderly donors].
This retrospective, case-controlled study compared the outcome of 17 OLTs (group A) using livers donated by subjects over 70 years of age with 17 OLTs (group B) with livers from donors under 40. Clinical data were used form the period 1996-1998. The following variables were considered in the analysis: donor clinical and laboratory parameters, cold ischemic periods, intra-operative blood and plasma replacement, 30-day mortality rate, incidence of primary graft dysfunction, acute rejection and arterial complication and long term survival. The main post-operative parameters were also included. Liver biopsy, performed in 9/17 of group A, revealed minimal steatosis. There were 2 post-operative deaths in group A and 1 in group B (p = NS). Two arterial complications were observed in group A (p = NS) and only one patient required retransplantation (p = NS). The only other difference found among clinical variables was the amount of total bilirubin at post-operative days 8 and 10, aPTT at days 6 and 13 and albumin at days 5 and 6. A two-year follow-up showed survival rates to be 88.2% and 94.1% for groups A and B, respectively (p = NS). Candidates over 70 years of age should be excluded as liver donors. In such cases, greater care needs to be placed on pathological vascular conditions related to advanced stage atherosclerosis such as calcified plaques on the hepatic artery, a possible factor in severe postoperative complications.


Mammographic breast density response to aromatase inhibition.
Mammographic breast density (MBD) is decreased by tamoxifen, but the effect of aromatase inhibitors is less clear. We enrolled early-stage postmenopausal patients with breast cancer initiating adjuvant aromatase inhibitor therapy and ascertained mammograms before and at an average 10 months of aromatase inhibitor therapy. We matched cases to healthy postmenopausal women (controls) from a large mammography screening cohort on age, baseline body mass index, baseline MBD, and interval between mammograms. We estimated change in MBD using a computer-assisted thresholding program (Cumulus) and compared differences between cases and matched controls. In predominantly White women (96%), we found 14% of the 387 eligible cases had a MBD reduction of at least 5% after an average of 10 months of aromatase inhibitor therapy. MBD reductions were associated with higher baseline MBD, aromatase inhibitor use for more than 12 months, and prior postmenopausal hormone use. Comparing each case with her matched control, there was no evidence of an association of change in MBD with aromatase inhibitor therapy [median case-control difference among 369 pairs was -0.1% (10th and 90th percentile: -5.9%, 5.2%) P = 0.51]. Case-control differences were similar by type of aromatase inhibitor (P's 0.41 and 0.56); prior use of postmenopausal hormones (P = 0.85); baseline MBD (P = 0.55); and length of aromatase inhibitor therapy (P = 0.08). In postmenopausal women treated with aromatase inhibitors, 14% of cases had a MBD reduction of more than 5%, but these decreases did not differ from matched controls. These data suggest that MBD is not a clinically useful biomarker for predicting the value of aromatase inhibitor therapy in White postmenopausal women.


Elective cardioversion: influence of paddle-electrode location and size on success rates and energy requirements.
We compared the success rates and energy requirements of two electrode-paddle positions (anteroposterior vs. anterolateral) and different paddle sizes in the elective cardioversion of atrial arrhythmias. We prospectively studied 173 patients - 111 in atrial fibrillation and 62 in atrial flutter. The anterolateral paddles used were either two standard-size (8.5-cm diameter) paddles or one 13-cm diameter anterior paddle with one standard-size lateral paddle. The anteroposterior paddles used were either a standard-size or a 13-cm anterior paddle with 12-cm posterior paddle. Overall cardioversion success rates with either paddle position were similar (greater than 90 per cent). The larger paddles did not significantly reduce energy requirements for cardioversion of either arrhythmia. We conclude that anterolateral paddles are as effective as anteroposterior paddles for the elective cardioversion of atrial arrhythmias, and that there is no demonstrable advantage to using paddles that are larger than the standard size in either position.


Dissociation between transmissible spongiform encephalopathy (TSE) infectivity and proteinase K-resistant PrP(Sc) levels in peripheral tissue from a murine transgenic model of TSE disease.
Most current diagnostic tests for transmissible spongiform encephalopathies (TSE) rely on the presence of proteinase K (PK)-resistant PrP(Sc) (PrP-res) in postmortem tissues as an indication of TSE disease. However, a number of studies have highlighted a discrepancy between TSE infectivity and PrP-res levels in both natural and experimental cases of TSE disease. Previously, we have shown high TSE infectivity levels in the brain tissue of mice that have a clinical TSE disease with associated vacuolar pathology but little or no detectable PrP-res. Here, the levels of TSE infectivity and PrP-res within a peripheral tissue of this mouse model were investigated. Biochemical analysis showed that low levels of PrP-res were present in the spleen tissue in comparison to the levels observed in the spleen of mice infected with ME7 or 79A. However, upon subpassage of brain and spleen tissue from clinically ill mice with little or no PrP-res detectable, similar short incubation periods to disease were observed, indicating that infectivity levels were similarly high in both tissues. Thus, the discrepancy between PrP-res and TSE infectivity was also present in the peripheral tissues of this disease model. This result indicates that peripheral tissues can contain higher levels of infectivity given the correct combination of host species, PrP genotype, and TSE agent. Therefore, the assumption that the levels of peripheral infectivity are lower than those in the central nervous system is not always correct, and this could have implications for current food safety regulations.


The toxicity of organoarsenic-based warfare agents: in vitro and in vivo studies.
The four arsenic-containing chemical warfare agents (CWA) Adamsite (technical, 10-chloro-9-10-dihydrophenarsazine), Clark 1 (Diphenyl arsine chloride), Clark 2 (Diphenyl arsine cyanide), and Lewisite (2-chloro-ethenyl dichloro arsine) were evaluated toxicologically using an in vitro and an in vivo model. The CWA were tested in vitro, using human leucocytes, in order to evaluate their effects on cell proliferation and cell cycle kinetics assayed by flow cytometry. The concentration for total inhibition of cell proliferation for the CWAs ranged from 0.3 micrograms/mL (Lewisite) to 15 micrograms/mL (Clark 1). The results showed no differences in cell proliferation between the different stages of the S-phase either at no-effect or at effect concentrations when samples treated with the CWAs were compared with untreated controls. As2O5 was used as a reference, and the arsenic concentration required for total inhibition of the cell proliferation was 1.7 mg/mL. A lower arsenic concentration, 0.83 micrograms/mL, showed a decreasing proliferation ration in the different parts of the S-phase with Searly and Slate having the highest and lowest ratios, respectively. In addition, there was a positive correlation between the unlabeled S-phase cell ratio and the arsenic concentration, indicating cessation of DNA-synthesis. Conclusively, the examined CWAs exerts a toxicity more potent than arsenic, with respect to cell proliferation. This might indicate that the toxicity of these arsenic-containing CWAs only to a minor extent can be explained by their arsenic content. Because of their relatively good water solubility and ability to easily degrade to arsenic acid, organo-arsenics, e.g., Lewisite, pose less of an environmental threat than organo-arsenics that are sparingly soluble and chemically persistent, like Adamsite. The four CWAs were also tested in vivo in a dietary exposure study, using juvenile three-spined stickleback (Gasterosteus aculeatus L.) as test organism, with measurement of EROD-activity and studies of liver morphology. The results from the in vivo study indicated small effects, suggesting that a 10-week period of dietary exposure at tested dose levels (1 and 100 ng/ml) affects juvenile three-spined stickleback only to a minor extent.


Toward a dynamical structure of DNA: comparison of theoretical and experimental NOE intensities.
Comparisons of experimental and calculated interproton nuclear Overhauser effect (NOE) buildup curves for duplex d(CGCGAATTCGCG)2 have been made. The calculated NOEs are based on molecular dynamics simulations including counterions and water and on the single-structure canonical A, B, and crystal forms. The calculated NOE effects include consideration of the motions of individual interproton vectors and the anisotropic tumbling of the DNA. The effects due to inclusion of anisotropic tumbling are much larger than those due to the local motion, and both improve the agreement between calculated and experimental results. The predictions based on the dynamical models agree significantly better with experiment than those based on either of the canonical forms or the crystal structure.


Development of a high fidelity pressurized porcine beating heart simulator for cardiac surgery training.
Development of a high-fidelity cardiac surgery simulator (CSS) requires integration of a heart model with a mock cardiopulmonary bypass (CPB) circuit that can provide feedback to mimic the pathophysiology of cardiac surgery. However, the cost of commercially available simulators precludes regular use. We describe steps in the construction of a high-fidelity CSS that integrates a pulsatile paracorporeal ventricular-assist device (Pulse-VAD) and a commercially available CPB simulator. Eight porcine hearts were initially prepared. The configuration consisted of cannulation of the distal descending aorta and the inferior vena cava to enable pressurization of the heart after connection to the Califia® simulator, as well as Pulse-VAD cannulation (fitted with inflatable balloons) of both ventricles. After each simulation run, the team addressed key issues to derive successive model changes through consensus. Key modifications included: a) pressure maintenance of the cardiac chambers (removal of lungs, Pulse-VAD cannulation sites at the left pulmonary artery and vein, double ligation of arch vessels); b) high-fidelity beating of both ventricles (full Pulse-VAD bladder filling and ensuring balloon neck placement at the valvular plane) and c) reproducible management of porcine anatomy (management of porcine aorta, ligation of left azygous vein and shortened ascending thoracic aortic segment). A CSS can be prepared at low cost, with integration into a high-fidelity CPB simulator with a novel beating heart component. This setup can be used in teaching the basics of CPB techniques and complex surgical procedures. Future work is needed to validate this model as a simulation instrument.


[Oxygen transport and hemodynamic reactions in ischemic heart disease patients when reserving and reinfusing massive volumes of autoblood].
The prospective randomized investigation was carried out in 61 patients with ischemic heart disease who were subjected to planned operations of coronary shunting under conditions of extracorporeal circulation. In the first group of patients autoblood as much as 24% of the calculated circulating blood volume (CBV) was reserved at the stage of isolation of the mammary artery, before systemic heparinization. The volume of the reserved blood of the patients of the second group was about 12% of CBV. It was shown that the reserving and reinfusion of massive (about 24% of CBV) volumes of autoblood in surgical treatment of ischemic heart disease under conditions of extracorporeal circulation failed to cause a substantial change of indices of central hemodynamics as compared to the reserving and reinfusion of autoblood in volume of about 12% of CBV.


T Scores, FRAX, Frailty Phenotype, Falls, and Its Relationship to Fractures in Patients on Maintenance Hemodialysis.
Despite the magnitude of fracture and the consequences in patients receiving hemodialysis, optimal risk assessment tools in this population are not well explored. Frailty and falls-known risk factors for fracture in chronic kidney disease (CKD) and non-CKD populations-are common in patients receiving hemodialysis (HD) therapy. While the relationship between T scores in relation to fractures in patients receiving HD is recognized, there is a paucity of data to the additional contributions of fracture assessment tool (FRAX), frailty status, and falls in its relationship with fracture. To evaluate the clinical utility of adding FRAX, frailty status, and falls to T scores at the femoral neck to determine whether it enhances fracture discrimination in patients on maintenance HD. A cross-sectional observational study. Two main dialysis units in Regina, Saskatchewan, Canada. A total of 109 patients on maintenance HD at two dialysis units from January 1, 2017, to December 31, 2018, were included in the study. Fracture (the main outcome) was documented based on the review of medical charts, self-recall, and additionally vertebral fractures were identified by an x-ray. Areal bone mineral density (BMD) was measured by dual-energy x-ray absorptiometry (DXA). FRAX score was calculated using an online algorithm based on 11 clinical risk factors. We calculated the FRAX score for hip fracture and major osteoprotoic fracture with and without the inclusion of BMD. Frailty was assessed using the Fried criteria, which included assessments of unintentional weight loss, weakness (handgrip strength), slowness (walking speed), and questionnaires for physical activity and self-perceived exhaustion. Patients were enquired about the history and frequency of falls. A total of 131 patients underwent frailty assessments at the two dialysis units during the dialysis treatment. Following frailty assessments, they were referred for DXA scans and upon receipt of the results undertook FRAX questionnaires. They were additionally sent for lumbar x-rays and contacted for a history of falls. Association between the BMD-T score, FRAX, frailty status, falls, with fracture were examined with sequential multivariable logistic regression models. Differences were considered statistically significant at P values <.05. A total of 109 patients were included in the data analysis. The composite of fracture occurred in 37.6% of patients. About 59.3% were identified as frail, and 29% of the participants had at least one fall in the last year. On multivariate regression analysis, each lower standard deviation (SD) in femoral neck T score was associated with 48% higher odds of fracture (odds ratio [OR] = 1.48; 95% confidence interval [CI] 1.20-1.68, P = .005). With the inclusion for FRAX scores (hip), the OR for fracture remained significant at 1.38 (OR = 1.38, 95% CI 1.04-1.63, P = .043). The addition of frailty status and history of falls did not further improve the model. Low T score and FRAX were both independent risk factors in patients on HD therapy. This is a single-center study with a small sample size which limits the generalizability of the findings. Due to the cross-sectional study, associations identified may be difficult to interpret. Both BMD measurements by DXA and FRAX are useful tools to assess fracture in patients receiving HD. The addition of frailty status and history of falls is not associated with fractures in this population. Larger prospective studies are needed to determine whether the inclusion of frailty and falls to the conventional models will improve fracture assessment in the population receiving HD. The study was not registered on a publicly accessible registry as it did not involve health care intervention on human participants.


Cromolyn sodium prophylaxis inhibits pulmonary proinflammatory cytokines in infants at high risk for bronchopulmonary dysplasia.
An imbalance of proinflammatory cytokines such as TNF-alpha, IL-1 beta, and the neutrophil chemotactic factor IL-8 and inhibitors (e.g., soluble TNF receptors and IL-1ra) in the lung during the first week of life may contribute to prolonged pulmonary inflammation and fibrosis in bronchopulmonary dysplasia (BPD). Disodium cromoglycate (DSCG) has anti-inflammatory effects in asthma, a disease with many similarities with BPD. In a prospective, randomized, blinded study, we examined whether early DSCG therapy inhibits proinflammatory cytokines in infants at risk for BPD. Twenty-six infants who were identified as high risk (> or = 75% probability) for oxygen-dependency at 28 d by a 12-h predictive score and survived 48 h were randomized to nebulized DSCG 20 mg (n = 13) or 2 cc NS (control, n = 13) every 6 h from Day 3 to Day 28. Lung lavage was collected on Day 3 (pre-study) and Day 7 and analyzed for cell count and differential and TNF-alpha, sTNFR1, sTNFR2, IL-1 beta, IL-1ra, and IL-8 concentrations. The groups' pre-study lavage cytokine concentrations were similar, but TNF-alpha and IL-8 concentrations were 3.6- and 4.9-fold lower in the DSCG group on Day 7 compared with levels in the control group. Soluble TNF receptors were unaffected by DSCG. There was a trend towards lower IL-1 beta levels in DSCG-treated infants on Day 7, but IL-1ra levels were unaffected by DSCG therapy. Three control subjects, but no DSCG-treated infants, died during the study period (p = 0.07). There were no significant differences between survivors of the two groups for oxygen-dependency at 28 d (100% control subjects; 85% DSCG). These results suggest that nebulized DSCG may exert an anti-inflammatory effect in the lungs of infants < or = 1,000 g at risk for BPD.


Polycomb complexes act redundantly to repress genomic repeats and genes.
Polycomb complexes establish chromatin modifications for maintaining gene repression and are essential for embryonic development in mice. Here we use pluripotent embryonic stem (ES) cells to demonstrate an unexpected redundancy between Polycomb-repressive complex 1 (PRC1) and PRC2 during the formation of differentiated cells. ES cells lacking the function of either PRC1 or PRC2 can differentiate into cells of the three germ layers, whereas simultaneous loss of PRC1 and PRC2 abrogates differentiation. On the molecular level, the differentiation defect is caused by the derepression of a set of genes that is redundantly repressed by PRC1 and PRC2 in ES cells. Furthermore, we find that genomic repeats are Polycomb targets and show that, in the absence of Polycomb complexes, endogenous murine leukemia virus elements can mobilize. This indicates a contribution of the Polycomb group system to the defense against parasitic DNA, and a potential role of genomic repeats in Polycomb-mediated gene regulation.


The effective concentration and exposure time of mitomycin-C for the inhibition of lens epithelial cell proliferation in rabbit eyes.
The proliferation of residual lens epithelial cells following cataract surgery is assumed to be a major cause of posterior capsular opacification. To assess the efficacy of mitomycin-C in preventing posterior capsular opacification, we determined the effective concentration and exposure time of mitomycin-C in inhibiting rabbit lens epithelial cell proliferation. The fourth-passaged rabbit lens epithelial cells were maintained for one day and then exposed to mitomycin-C for 1, 2, 3, and 5 minutes, respectively. There were 9 different plating concentrations of mitomycin-C with two-fold serial dilution. The maintenance of the phenotypic properties of lens epithelial cells was confirmed by continuous transcription of lambda-crystalline mRNA determined by reverse transcription-polymerase chain reaction and the polymorphism of the restriction fragment. Cell proliferation was assayed with 3H-thymidine incorporation into DNA. The fourth-passaged cells maintained the expression of lambda-crystalline mRNA, suggesting that they are phenotypically authentic lens epithelial cells. The effective concentrations and exposure time of mitomycin-C were 0.1 mg/ml for 1 minute and 2 minutes, and 0.025 mg/ml for 2 minutes. By these results, we postulated that mitomycin-C at relatively short incubation times could be clinically used for prevention of posterior capsular opacification after cataract surgery.


Low chance of survival among patients requiring adrenaline (epinephrine) or intubation after out-of-hospital cardiac arrest in Sweden.
To relate the outcome of out-of-hospital cardiac arrest to whether medication with adrenaline (epinephrine) was given and whether patients were intubated. A national survey in Sweden between 1990-1995 among patients suffering out-of-hospital cardiac arrest and in whom resuscitation was attempted. Sixty per cent of ambulance organisations in Sweden participated. Prospective evaluation. Survival was defined as survival 1 month after cardiac arrest. In all, 14065 patients were included in the evaluation. Of these, resuscitation was attempted in 10966 cases. Among these adrenaline (epinephrine) was given in 42.4 and 47.5% were intubated. In an univariate analysis treatment with adrenaline (epinephrine) and intubation was associated with a lower survival when all patients were evaluated. In a multivariate analysis including age, sex, place of arrest, bystander-CPR, initial arrhythmia, arrest being witnessed and aetiology, treatment with adrenaline (epinephrine) (OR 0.43, CI 0.27-0.66) and intubation (OR 0.71, CI 0.51-0.99) were both independent predictors of a lower chance of survival. When separately analysing patients with bystander witnessed cardiac arrest found in ventricular fibrillation and requiring more than 3 defibrillatory shocks neither treatment with adrenaline (epinephrine) nor intubation was associated with survival. Among patients with a non-shockable rhythm treatment with adrenaline (epinephrine) was a significant independent predictor for lower survival (OR 0.30, CI 0.07-0.82). In a national survey in Sweden including 10966 cases of out-of-hospital cardiac arrest the outcome was related to whether medication with adrenaline (epinephrine) was given and whether patients were intubated. Neither in total nor in any subgroup did we find results indicating beneficial effects of any of these two interventions. Whether treatment with adrenaline (epinephrine) or intubation will increase survival after out-of-hospital cardiac arrest needs to be confirmed in prospective randomised trials.


Estimating the growth potential of the soil protozoan community.
We have developed a method for determining the potential abundance of free-living protozoa in soil. The method permits enumeration of four major functional groups (flagellates, naked amoebae, testate amoebae, and ciliates) and it overcomes some limitations and problems of the usual 'direct' and 'most probable number' methods. Potential abundance is determined using light microscopy, at specific time intervals, after quantitative re-wetting of air-dried soil with rain water. No exogenous carbon substrates or mineral nutrients are employed, so the protozoan community that develops is a function of the resources and inhibitors present in the original field sample. The method was applied to 100 soil samples (25 plots x 4 seasons) from an upland grassland (Sourhope, Southern Scotland) in the UK. Median abundances for all four functional groups lie close to those derived from the literature on protozoa living in diverse soil types. Flagellates are the most abundant group in soil, followed by the naked amoebae, then the testate amoebae and ciliates. This order is inversely related to typical organism size in each group. Moreover, preliminary evidence indicates that each functional group contains roughly the same number of species. All of these observations would be consistent with soil having fractal structure across the size-scale perceived by protozoa. The method described will be useful for comparing the effects on the soil protozoan community of different soil treatments (e.g. liming and biocides).


Estrous cycle influences organ-specific metastasis of B16F10 melanoma cells.
Some clinical studies suggest that timing of surgery during specific menstrual phases may influence the chances of survival for premenopausal women with breast cancer, whereas other studies failed to find this effect. Because most breast cancer deaths are attributable to metastases, we hypothesized that aspects of the metastatic process might be sensitive to cyclic hormonal fluctuations. Our goal was to develop a mouse model to assess possible mechanisms for the effect of the menstrual cycle on metastatic ability. To separate the effects of the hormonal milieu on the host versus the cancer cells, we began by using melanoma cells. We report here that the estrous phase at the time of entry of B16F10 melanoma cells into the circulation leads to marked differences in organ-specific metastasis, suggesting that this concept merits additional study.


Outcome of Patients with Complete Unilateral Cleft Lip and Palate: 20-Year Follow-Up of a Treatment Protocol.
The treatment plan for cleft lip and palate varies among centers and requires long-term evaluation of its final outcome. A consecutive series of patients born from 1994 to 1996 were reviewed. Inclusion criteria were complete unilateral cleft lip and palate, undergoing all treatment procedures performed by the team, and continuous follow-ups until 20 years of age. Exclusion criteria were incomplete data, having microform cleft lip on the contralateral side, presence of the Simonart band, and other abnormalities. A total of 72 patients were included. Average age at final evaluation was 21.3 years; 83.3 percent of patients underwent one-stage rotation-advancement lip repair and 16.7 percent underwent two-stage repair with an initial adhesion cheiloplasty. All patients underwent palate repair using the two-flap method at an average age of 12.3 months. Velopharyngeal insufficiency occurred and required surgical interventions in 19.4 percent during the preschool age and in 16.7 percent at the time of alveolar bone grafting; 56.9 percent of patients underwent secondary lip/nose revision during the growing age. Regular orthodontic treatment was administered to 34.7 percent of patients between 12 and 16 years of age. Orthodontic treatment and orthognathic surgery were applied in 37.5 percent of the patients after maturity. The average number of surgical procedures to complete the treatment was 4.8 per patient. This treatment protocol provided generally acceptable final outcome after the 20-year follow-up. Some results were less ideal and have resulted in modifications of the planning and methods in the protocol. Therapeutic, IV.


ROS-related redox regulation and signaling in plants.
As sessile oxygenic organisms with a plastic developmental programme, plants are uniquely positioned to exploit reactive oxygen species (ROS) as powerful signals. Plants harbor numerous ROS-generating pathways, and these oxidants and related redox-active compounds have become tightly embedded into plant function and development during the course of evolution. One dominant view of ROS-removing systems sees them as beneficial antioxidants battling to keep damaging ROS below dangerous levels. However, it is now established that ROS are a necessary part of subcellular and intercellular communication in plants and that some of their signaling functions require ROS-metabolizing systems. For these reasons, it is suggested that "ROS processing systems" would be a more accurate term than "antioxidative systems" to describe cellular components that are most likely to interact with ROS and, in doing so, transmit oxidative signals. Within this framework, our update provides an overview of the complexity and compartmentation of ROS production and removal. We place particular emphasis on the importance of ROS-interacting systems such as the complex cellular thiol network in the redox regulation of phytohormone signaling pathways that are crucial for plant development and defense against external threats.


Drug-alcohol interactions on psychomotor skills: zopiclone and flunitrazepam.
Interaction between alcohol and zopiclone or flunitrazepam as well as the residual effects of both these hypnotics were studied in 20 normal male volunteers who received each 3 of 6 different drug and drink combinations according to a balanced incomplete block design as follows: placebo, zopiclone (7.5 mg), or flunitrazepam (2 mg) was administered double-blind in identical capsules at 23.00 h, and 0.5 g/kg body weight of alcohol or placebo alcohol was given at 08.30 h the following morning. Psychomotor skills of the volunteers were measured before drinking and 30 min, 1.5 h and 2.5 h after it. As compared with placebo, zopiclone had no residual effects, while flunitrazepam had some effects on standing steadiness, tracking, and flicker recognition. Alcohol alone had a non-significant effect on skills, and the combination zopiclone plus alcohol behaved as alcohol alone. Flunitrazepam plus alcohol impaired significantly standing steadiness, tracking, and reactive skills when compared with all other treatments. Time anticipation and hand and foot proprioception were not affected by any treatment. The results suggest that flunitrazepam, unlike zopiclone, has residual effects and interacts with alcohol in the morning following overnight ingestion of the drug.


Novel mutation in the SPG3A gene in an African American family with an early onset of hereditary spastic paraplegia.
Mutations in a novel GTPase gene SPG3A cause an autosomal dominant hereditary spastic paraplegia linked to chromosome 14q (SPG3), which accounts for approximately 10% to 15% of all autosomal dominant hereditary spastic paraplegia cases. The mutational spectrum of the SPG3A gene and the phenotype/genotype correlations have not yet been established. To describe a kindred with an infantile onset of hereditary spastic paraplegia caused by a novel mutation in the SPG3A gene. Complete neurological examination and genetic analysis were performed on 6 affected members of a small African American kindred. Linkage analysis to genetic markers near autosomal dominant hereditary spastic paraplegia loci on chromosomes 2p and 14q was performed. The coding sequence of the SPG3A gene was analyzed, and the identified change in the sequence was tested for being a benign polymorphism by sequencing 200 chromosomes from normal controls. Every affected individual had signs of uncomplicated spastic paraparesis without additional neurological abnormalities. None of the affected family members had ever walked normally. The history was consistent with an infantile onset, despite the normal acquisition of motor milestones. Genetic analysis suggested linkage to the SPG3A locus on chromosome 14q. Analysis of the SPG3A gene revealed a missense mutation C635T, predicted to result in a threonine to isoleucine substitution at codon 156. Analysis of 200 normal chromosomes did not identify the same change in healthy subjects. We report a novel mutation in the SPG3A gene in an African American family with an infantile onset of autosomal dominant hereditary spastic paraplegia.


Managing reduced methotrexate clearance in a patient with a heterozygous methylenetetrahydrofolate reductase gene polymorphism.
High dose methotrexate has become one of the treatments of choice for patients with primary CNS lymphomas due to its ability to penetrate the blood-brain barrier. A potentially serious complication of this therapy is methotrexate-related nephrotoxicity. We report the case of a patient with a common genetic polymorphism that may have predisposed this patient experience clinically significant toxicity from systemic folate depletion. After the first cycle of chemotherapy that included high dose methotrexate, the patient's serum creatinine rose and the patient's methotrexate level remained above the toxic range for six days. On cycle two, the patient was treated with a 25% dose reduction in methotrexate and more aggressive hydration and alkalization. With this alteration in the regimen, the patient was able to receive six more cycles and had a complete radiographic tumor response in the brain and a disappearance of tumor cells in the CSF without any further renal complications. This case report illustrates the feasibility of administering high dose methotrexate with modifications as a treatment of choice in individuals with methylenetetrahydrofolate reductase gene mutations.


The impact of group B Streptococcus prophylaxis on early onset neonatal infections.
To determine if changes have occurred in the causative pathogens and/or antibiotic susceptibility profiles in early onset neonatal infections since initiation of group B Streptococcus (GBS) prophylaxis and to determine risk factors for ampicillin/penicillin resistant microorganisms. Data on 220 infants with positive blood, urine, or cerebrospinal fluid cultures for bacteria or fungi at ≤seven days of age from 1990-2007 were examined and divided into three epochs, based on intrapartum antibiotic prophylactic (IAP) practices. Pathogens and antibiotic resistance were compared among epochs. A significant decrease in the incidence of GBS infections occurred over time, with no change in the incidence of other pathogens or the emergence of antibiotic resistance, including the very low-birthweight population. In regression analysis, ampicillin resistance was associated with male gender (OR 3.096). No emergence of antibiotic resistant pathogens was found following IAP use. Changing microorganisms and increasing antibiotic resistance found in prior studies are likely multifactorial. Further study is needed to continue to reduce the rates of common early onset pathogens.


Lipoteichoic acid enhances IL-6 production in human synovial fibroblasts via TLR2 receptor, PKCdelta and c-Src dependent pathways.
Patients with rheumatoid arthritis (RA) are at increased risk of developing infections and appear to be particularly susceptible to septic arthritis. Lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria is an amphiphilic, negatively charged glycolipid. However, the effects of LTA on human synovial fibroblasts are largely unknown. We investigated the signaling pathway involved in IL-6 production stimulated by LTA in rheumatoid arthritis synovial fibroblasts (RASF). LTA caused concentration- and time-dependent increases in IL-6 production. LTA-mediated IL-6 production was attenuated by Toll-like receptor 2 (TLR2) monoclonal antibody or siRNA. Pretreatment with PKCdelta inhibitor (rottlerin), c-Src inhibitor (PP2), AP-1 inhibitor (tanshinone IIA) and NF-kappaB inhibitor (PDTC and TPCK) also inhibited the potentiating action of LTA. However, focal adhesion kinase (FAK) mutant and siRNA did not affect LTA-mediated IL-6 production. Stimulation of cells with LTA increased the PKCdelta and c-Src phosphorylation and kinase activity. LTA increased the accumulation of p-c-Jun and p-p65 in the nucleus, as well as AP-1 and NF-kappaB luciferase activity. LTA-mediated increase of AP-1 and NF-kappaB luciferase activity was inhibited by rottlerin and PP2 or TLR2 and PKCdelta siRNA or c-Src mutant. Our results suggest that LTA-increased IL-6 production in human synovial fibroblasts via the TLR2 receptor, PKCdelta, c-Src, AP-1 and NF-kappaB signaling pathways.


Preparation of polymer gold nanoparticle composites with tunable plasmon coupling and their application as SERS substrates.
The controlled surface functionalisation of polystyrene beads (200 nm) with a lipoic acid derivative is used to assemble composites with between 4 to 20% loadings of citrate stabilised gold nanoparticles (13 nm-30 nm), which exhibit variable optical properties arising from interactions of the nanoparticle surface plasmon resonance (SPR). The decrease in average interparticle distance at higher loadings results in a red-shift in the SPR wavelength, which is well described by a universal ruler equation. The composite particles are shown to act as good SERS substrates for the standard analyte 4-mercaptophenol. The direct assessment of the SERS activity for individual composite particles solution is achieved by Raman optical tweezer measurements on 5.3 μm composite particles. These measurements show an increase in performance with increasing AuNP size. Importantly, the SERS activity of the individual particles compares well with the bulk measurements of samples deposited on a surface, indicating that the SERS activity arises primarily from the composite and not due to composite-composite interactions. In both studies the optimum SERS response is obtained with 30 nm AuNPs.


Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential.
We previously found that human fetal lung is a rich source of mesenchymal stem cells (MSC). Here we characterize and analyze the frequency and function of MSC in other second-trimester fetal tissues. Single cell suspensions of fetal bone marrow (BM), liver, lung, and spleen were made and analyzed by flow cytometry for the expression of CD90, CD105, CD166, SH3, SH4, HLA-ABC, HLA-DR, CD34 and CD45. We assessed the frequency of MSC by limiting dilution assay. The frequency of MSC in BM was significantly higher than in liver, lung, and spleen (p<0.05). On primary non-expanded cells from fetal liver, lung and spleen the number of cells positive for mesenchymal markers was significantly higher within the CD34 positive population than within the CD34 negative population. The phenotype of the culture-expanded MSC was similar for all fetal tissues, i.e. CD90, CD105, CD166, SH3, SH4 and HLA-ABC positive and CD34, CD45 and HLA-DR negative. Culture-expanded cells from all tissues were able to differentiate along adipogenic and osteogenic pathways. However, adipogenic differentiation was less in MSC derived from spleen, and osteogenic differentiation was reduced in liver-derived MSC (p<0.05). Our results indicate that culture-expanded MSC derived from second-trimester fetal tissues, although phenotypically similar, exhibit heterogeneity in differentiating potential. We speculate that these differences may be relevant for the clinical application of MSC.


Pazufloxacin Toyama Chemical Co.
Toyama Chemical Co Ltd is developing pazufloxacin (T-3761), an orally active synthetic quinolone antibiotic, which is awaiting registration following successful clinical trials that demonstrated its pharmacological similarities to tosufloxacin (Toyama Chemical Co) with respect to clinical efficacy, adverse effects and overall safety [272136]. As of June 2000, the company had filed an NDA for marketing approval in Japan [373027]. The company is also developing an injectable formulation, pazufloxacin mesylate (T-3762), which has entered phase III trials in Japan [228819]. There is a licensing agreement with Welfide Corp (formerly Yoshitomi/The Green Cross Corp) in Japan [228819].


[Localization of Ascaris lumbricoides in the thoracic cavity. Report of a case].
Several aspects of the history, evolutive cycle and erratic phases of Ascaris lumbricoides as well as of the symptoms and diagnosis of ascariasis are analyzed. The anatomicophysiological concept of spontaneous pneumothorax and its clinical picture, frequency and treatment are exposed. A patient who underwent thoracic surgery due to an acute respiratory syndrome in who a male Ascaris lumbricoides was surprisingly found obstructing the probe after an explorating left thoracotomy and a resection of the lung apex is presented. The nematode is suggested as the possible etiological agent of the respiratory disease.


Cardiac Amyloidosis: Evolving Diagnosis and Management: A Scientific Statement From the American Heart Association.
Transthyretin amyloid cardiomyopathy (ATTR-CM) results in a restrictive cardiomyopathy caused by extracellular deposition of transthyretin, normally involved in the transportation of the hormone thyroxine and retinol-binding protein, in the myocardium. Enthusiasm about ATTR-CM has grown as a result of 3 simultaneous areas of advancement: Imaging techniques allow accurate noninvasive diagnosis of ATTR-CM without the need for confirmatory endomyocardial biopsies; observational studies indicate that the diagnosis of ATTR-CM may be underrecognized in a significant proportion of patients with heart failure; and on the basis of elucidation of the mechanisms of amyloid formation, therapies are now approved for treatment of ATTR-CM. Because therapy for ATTR-CM may be most effective when administered before significant cardiac dysfunction, early identification of affected individuals with readily available noninvasive tests is essential. This scientific statement is intended to guide clinical practice and to facilitate management conformity by covering current diagnostic and treatment strategies, as well as unmet needs and areas of active investigation in ATTR-CM.


Neurovascular coupling and cerebral autoregulation in atrial fibrillation.
The risk of cognitive decline and stroke is increased by atrial fibrillation (AF). We sought to determine whether neurovascular coupling and cerebral autoregulation are blunted in people with AF in comparison with age-matched, patients with hypertension and healthy controls. Neurovascular coupling was assessed using five cycles of visual stimulation for 30 s followed by 30 s with both eyes-closed. Cerebral autoregulation was examined using a sit-stand test, and a repeated squat-to-stand (0.1 Hz) manoeuvre with transfer function analysis of mean arterial pressure (MAP; input) and middle cerebral artery mean blood flow velocity (MCA Vm; output) relationships at 0.1 Hz. Visual stimulation increased posterior cerebral artery conductance, but the magnitude of the response was blunted in patients with AF (18 [8] %; mean [SD]) and hypertension (17 [8] %), in comparison with healthy controls (26 [9] %) (P < 0.05). In contrast, transmission of MAP to MCA Vm was greater in AF patients compared to hypertension and healthy controls, indicating diminished cerebral autoregulation. We have shown for the first time that AF patients have impaired neurovascular coupling responses to visual stimulation and diminished cerebral autoregulation. Such deficits in cerebrovascular regulation may contribute to the increased risk of cerebral dysfunction in people with AF.


[Mechanism of enzymatic imprinting induced in rats by an early postnatal administration of galactose].
Administration of galactose into young rats within an early postnatal period led to alteration in activity of some enzymes involved in utilization of galactose (galactose-1-phosphaturidyl transferase, galactose-6-phosphate dehydrogenase etc) for a long period of the animals life. This stable alteration in activity of adaptive enzymes was characterized as the enzymatic imprinting. After administration of galactose into neonatal animals synthesis of RNA, matrix activity of chromatin, activities of DNA-dependent RNA polymerase and RNA-dependent DNA polymerase were shown to increase in liver tissue of these animals. These alterations are considered as a possible basis for the stable alterations in the genes expression. The elevated activities of DNA-dependent RNA-polymerase and reverse transcriptase were maintained within a long period of the animals life.


Systematic review of percutaneous interventions for malignant pericardial effusion.
The present systematic review assessed the safety and efficacy of percutaneous interventions for malignant pericardial effusion (MPE), with primary endpoint of recurrence of pericardial effusion. Electronic searches of six databases identified thirty-one studies, reporting outcomes following isolated pericardiocentesis (n=305), pericardiocentesis followed by extended catheter drainage (n=486), pericardial instillation of sclerosing agents (n=392) or percutaneous balloon pericardiotomy (PBP) (n=157). Isolated pericardiocentesis demonstrated a pooled recurrence rate of 38.3%. Pooled recurrence rates for extended catheter drainage, pericardial sclerosis and PBP were 12.1%, 10.8% and 10.3%, respectively. Procedure-related mortality ranged from 0.5-1.0% across the percutaneous interventions. Although isolated pericardiocentesis can safely deliver immediate symptomatic relief, subsequent catheter drainage or sclerotherapy are required to minimize recurrence. PBP has been shown to be highly effective and may be particularly useful in managing recurrent effusions. Ultimately, the choice of intervention must be based on the clinical status of patients, their underlying malignancy and the expertise available.


User needs and requirements analysis for big data healthcare applications.
The realization of big data applications that allow improving the quality and efficiency of healthcare care delivery is challenging. In order to take advantage of the promising opportunities of big data technologies, a clear understanding of user needs and requirements of the various stakeholders of healthcare, such as patients, clinicians and physicians, healthcare provider, payors, pharmaceutical industry, medical product suppliers and government, is needed. Our study is based on internet, literature and market study research as well as on semi-structured interviews with major stakeholder groups of healthcare delivery settings. The analysis shows that big data technologies could be used to align the opposing user needs of improved quality with improved efficiency of care. However, this requires the integrated view of various heterogeneous data sources, legal frameworks for data sharing and incentives that foster collaboration.


PUSK AND PULL FACTORS OF EMIGRATION AMONG PHYSICIAN IN NIGERIA.
Physician emigration is increasing exponentially in developing countries. In Nigeria, with the last decade's unprecedented brain drain, it has gained the popular moniker 'japa syndrome'. This study aimed to determine push and pull factors affecting physician migration in Nigeria, to provide evidence-backed recommendations for physician retention policies. A cross-sectional study was conducted among attendees at the 2022 Abuja Cardiovascular Symposium hosted by Limi Multispecialty Hospital and the Nigerian Cardiac Society. Convenience and snowball sampling were used, and 295/400 responded to comprehensive self-administered questionnaires (73.7% response rate). Data was analysed using SPSS v.26. Most participants (79.4%) were aged 20-39 years (Mean 35 years SD ±10.17); female (58.6%); married (58.4%) and had family size below six (73.6%). About 85.8% were employed, and 55.9% worked in private establishments. Solely basic medical degrees were possessed by 64.4%, and 63.7% earned N300,000-N399,999 (USD 396.82-USD 527.78) monthly. Top destinations were UK (50.5%), Canada (43.3%), and USA (37.9%), with low remuneration (71.2%), insecurity (62.7%), and difficult working environments (55.9%) most frequent push factors. Postgraduate-training frustrations (38.6%), and limited educational opportunities for oneself (37.6%), children (26.4%), or spouse (19.7%) were the least. High earning potential (76.6%), career growth opportunities (70.8%), and high-level equipment/technology (54.9%) were frequent pull factors. Physician emigration threatens Nigeria's health system and should be addressed multi-sectorally to boost physician remuneration and improve work environments and societal security. Additionally, innovative education and digital technology would encourage health workforce retention.


Multiyear Measurements on Δ17O of Stream Nitrate Indicate High Nitrate Production in a Temperate Forest.
Nitrification is a crucial step in ecosystem nitrogen (N) cycling, but scaling up from plot-based measurements of gross nitrification to catchments is difficult. Here, we employed a newly developed method in which the oxygen isotope anomaly (Δ17O) of nitrate (NO3-) is used as a natural tracer to quantify in situ catchment-scale gross nitrification rate (GNR) for a temperate forest from 2014 to 2017 in northeastern China. The annual GNR ranged from 71 to 120 kg N ha-1 yr-1 (average 94 ± 10 kg N ha-1 yr-1) over the 4 years in this forest. This result and high stream NO3- loss (4.2-8.9 kg N ha-1 yr-1) suggest that the forested catchment may have been N-saturated. At the catchment scale, the total N output of 10.7 kg N ha-1 yr-1, via leaching and gaseous losses, accounts for 56% of the N input from bulk precipitation (19.2 kg N ha-1 yr-1). This result indicates that the forested catchment is still retaining a large fraction of N from atmospheric deposition. Our study suggests that estimating in situ catchment-scale GNR over several years when combined with other conventional flux estimates can facilitate the understanding of N biogeochemical cycling and changes in the ecosystem N status.


The highly conserved U small nuclear RNA 3'-end formation signal is quite tolerant to mutation.
Formation of the 3' end of U1 and U2 small nuclear RNA (snRNA) precursors is directed by a conserved sequence called the 3' box located 9 to 28 nucleotides downstream of all metazoan U1 to U4 snRNA genes sequenced so far. Deletion of part or all of the 3' box from human U1 and U2 genes drastically reduces 3'-end formation. To define the essential nucleotides within this box that direct 3'-end formation, we constructed a set of point mutations in the conserved residues of the human U1 3' box. The ability of the various mutations to direct 3'-end formation was tested by microinjection into Xenopus oocytes and transfection into HeLa cells. We found that the point mutations had diverse effects on 3'-end formation, ranging from no effect at all to severe inhibition; however, no single or double point mutation we tested completely eliminated 3'-end formation. We also showed that a rat U3 3' flank can effectively substitute for the human U1 3' flank, indicating that the 3' boxes of the different U snRNA genes are functionally equivalent.


Long-term mortality data from the balloon pump-assisted coronary intervention study (BCIS-1): a randomized, controlled trial of elective balloon counterpulsation during high-risk percutaneous coronary intervention.
There is conflicting evidence on the utility of elective intra-aortic balloon pump (IABP) use during high-risk percutaneous coronary intervention (PCI). Observational series have indicated a reduction in major in-hospital adverse events, although randomized trial evidence does not support this. A recent study has suggested a mortality benefit trend early after PCI, but there are currently no long-term outcome data from randomized trials in this setting. Three hundred one patients with left ventricular impairment (ejection fraction <30%) and severe coronary disease (BCIS-1 jeopardy score ≥8; maximum possible score=12) were randomized to receive PCI with elective IABP support (n=151) or without planned IABP support (n=150). Long-term all-cause mortality was assessed by tracking the databases held at the Office of National Statistics (in England and Wales) and the General Register Office (in Scotland). The groups were balanced in terms of baseline characteristics (left ventricular ejection fraction, 23.6%; BCIS-1 jeopardy score, 10.4) and the amount and type of revascularization performed. Mortality data were available for the entire cohort at a median of 51 months (interquartile range, 41-58) from randomization. All-cause mortality at follow-up was 33% in the overall cohort, with significantly fewer deaths occurring in the elective IABP group (n=42) than in the group that underwent PCI without planned IABP support (n=58) (hazard ratio, 0.66; 95% confidence interval, 0.44-0.98; P=0.039). In patients with severe ischemic cardiomyopathy treated with PCI, all-cause mortality was 33% at a median of 51 months. Elective IABP use during PCI was associated with a 34% relative reduction in all-cause mortality compared with unsupported PCI. URL: http://www.isrctn.org. Unique identifier: ISRCTN40553718; and http://www.clinicaltrials.gov. Unique identifier: NCT00910481.


Continuous-infusion diaziquone in acute myeloid leukemia: a Southeastern Cancer Study Group trial.
Diaziquone (AZQ) is a synthetic quinone with considerable activity against L1210 leukemia and potent myelosuppressive activity in man. To test the efficacy and toxicity of AZQ administered by continuous infusion, a phase II multi-institutional trial was undertaken by the Southeastern Cancer Study Group. Eligible adults with acute myeloid leukemia (AML) received AZQ at a dose of 28 mg/m2 daily by continuous infusion for 5 days. Patients failing to achieve complete remission received a second course utilizing the same dose and schedule. Of 25 evaluable patients with relapsed or refractory AML, three achieved complete response (12%) and two achieved partial response (8%). All patients experienced marked myelosuppression. Severe or life-threatening infection was observed in 15 (56%) patients. Clinical or postmortem evidence of central nervous system hemorrhage was encountered in three (12%) patients with severe refractory thrombocytopenia. Minimal nonhematologic toxicity was observed, suggesting that further studies of this agent in combination regimens and possibly for marrow transplantation preparation in patients with acute leukemia are warranted.


Circulating miRNAs as Diagnostic Biomarkers for Parkinson's Disease.
Parkinson's disease (PD) is the second most common neurodegenerative disorder worldwide. Its main neuropathological hallmarks are the degeneration of dopaminergic neurons in the substantia nigra and alpha-synuclein containing protein inclusions, called Lewy Bodies. The diagnosis of idiopathic PD is still based on the assessment of clinical criteria, leading to an insufficient diagnostic accuracy. Additionally, there is no biomarker available allowing the prediction of the disease course or monitoring the response to therapeutic approaches. So far, protein biomarker candidates such as alpha-synuclein have failed to improve diagnosis of PD. Circulating microRNAs (miRNAs) in body fluids are promising biomarker candidates for PD, as they are easily accessible by non- or minimally-invasive procedures and changes in their expression are associated with pathophysiological processes relevant for PD. Advances in miRNA analysis methods resulted in numerous recent publications on miRNAs as putative biomarkers. Here, we discuss the applicability of different body fluids as sources for miRNA biomarkers, highlight technical aspects of miRNA analysis and give an overview on published studies investigating circulating miRNAs as biomarker candidates for diagnosis of PD and other Parkinsonian syndromes.


The histone variant H2A.Z promotes splicing of weak introns.
Multiple lines of evidence implicate chromatin in the regulation of premessenger RNA (pre-mRNA) splicing. However, the influence of chromatin factors on cotranscriptional splice site usage remains unclear. Here we investigated the function of the highly conserved histone variant H2A.Z in pre-mRNA splicing using the intron-rich model yeast Schizosaccharomyces pombe Using epistatic miniarray profiles (EMAPs) to survey the genetic interaction landscape of the Swr1 nucleosome remodeling complex, which deposits H2A.Z, we uncovered evidence for functional interactions with components of the spliceosome. In support of these genetic connections, splicing-specific microarrays show that H2A.Z and the Swr1 ATPase are required during temperature stress for the efficient splicing of a subset of introns. Notably, affected introns are enriched for H2A.Z occupancy and more likely to contain nonconsensus splice sites. To test the significance of the latter correlation, we mutated the splice sites in an affected intron to consensus and found that this suppressed the requirement for H2A.Z in splicing of that intron. These data suggest that H2A.Z occupancy promotes cotranscriptional splicing of suboptimal introns that may otherwise be discarded via proofreading ATPases. Consistent with this model, we show that overexpression of splicing ATPase Prp16 suppresses both the growth and splicing defects seen in the absence of H2A.Z.