FimK regulation on the expression of type 1 fimbriae in Klebsiella pneumoniae CG43S3.
Klebsiella pneumoniae CG43, a heavy encapsulated liver abscess isolate, mainly expresses type 3 fimbriae. Type 1 fimbriae expression was only apparent in CG43S3ΔmrkA (the type 3 fimbriae-deficient strain). The expression of type 1 fimbriae in CG43S3ΔmrkA was reduced by deleting the fimK gene, but was unaffected by removing the 3' end of fimK encoding the C-terminal EIL domain (EILfimK). Quantitative RT-PCR and promoter activity analysis showed that the putative DNA-binding region at the N terminus, but not the C-terminal EIL domain, of FimK positively affects transcription of the type 1 fimbrial major subunit, fimA. An electrophoretic mobility shift assay demonstrated that the recombinant FimK could specifically bind to fimS, which is located upstream of fimA and contains a vegetative promoter for the fim operon, also reflecting possible transcriptional regulation. EILfimK was shown to encode a functional phosphodiesterase (PDE) via enhancing motility in Escherichia coli JM109 and in vitro using PDE activity assays. Moreover, EILfimK exhibited higher PDE activity than FimK, implying that the N-terminal DNA-binding domain may negatively affect the PDE activity of FimK. FimA expression was detected in CG43S3 expressing EILfimK or AILfimK, suggesting that FimA expression is not directly influenced by the c-di-GMP level. In summary, FimK influences type 1 fimbriation by binding to fimS at the N-terminal domain, and thereafter, the altered protein structure may activate C-terminal PDE activity to reduce the intracellular c-di-GMP level.


Twenty four hour manometric recording of colonic motor activity in healthy man.
The motor activity of the transverse, descending, and sigmoid colon was recorded for 24 hours in 14 healthy volunteers with a colonoscope positioned catheter. During the study the patients ate two 1000 kcal mixed meals and one continental breakfast. Colonic motor activity was low before meals and minimal during sleep; the motility index increased significantly after meals and at morning awakening. Most of the motor activity was represented by low amplitude contractions present singly or in bursts, which showed no recognisable pattern. All but two subjects also showed isolated high amplitude (up to 200 mmHg) contractions that propagated peristaltically over long distances at approximately 1 cm/sec. Most of these contractions occurred after morning awakening, and some in the late postprandial period, with a mean of 4.4/subject/24 h. The peristaltic contractions were often felt as an urge to defecate or preceded defecation, and could represent the manometric equivalent of the mass movements.


Fenugreek (Trigonella foenum graecum) seed polyphenols protect liver from alcohol toxicity: a role on hepatic detoxification system and apoptosis.
The present study investigates the hepatoprotective effect of fenugreek seed polyphenolic extract (FPEt) against ethanol-induced hepatic injury and apoptosis in rats. Chronic ethanol administration (6 g/kg/day x 60 days) caused liver damage that was manifested by the elevation of markers of liver dysfunction--aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), bilirubin and gamma-glutamyl transferase (GGT) in plasma and reduction in liver glycogen. The effects on alcohol metabolizing enzymes such as alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) were studied and found to be altered in the alcohol-treated group. Ethanol administration resulted in adaptive induction of the activities of cytochrome p450 (cyt-p-450) and cytochrome-b5 (cyt-b5) and reduction in cytochrome-c-reductase (cyt-c-red) and glutathione-S-tranferase (GST), a phase II enzyme. Further, ethanol reduced the viability of isolated hepatocytes (ex vivo) as assessed by the trypan blue exclusion test and increased hepatocyte apoptosis as assessed by propidium iodide staining (PI). Treatment with FPEt restored the levels of markers of liver injury and mitigated the alterations in alcohol metabolizing and detoxification enzymes and the electron transport component cytochrome-c reductase. Increased hepatocyte viability and reduced apoptotic nuclei were observed in FPEt-treated rats. These findings demonstrate that FPEt acts as a protective agent against ethanol-induced abnormalities in the liver. The effects of FPEt are comparable with those of a known hepatoprotective agent, silymarin.


A Surgical Procedure for the Administration of Drugs to the Inner Ear in a Non-Human Primate Common Marmoset (Callithrix jacchus).
Hearing research has long been facilitated by rodent models, although in some diseases, human symptoms cannot be recapitulated. The common marmoset (Callithrix jacchus) is a small, easy-to-handle New World monkey which has a similar anatomy of the temporal bone, including the middle ear ossicular chains and inner ear to humans, than in comparison with that of rodents. Here, we report a reproducible, safe, and rational surgical approach to the cochlear round window niche for the drug delivery to the inner ear of the common marmoset. We adopted posterior tympanotomy, a procedure used clinically in human surgery, to avoid manipulation of the tympanic membrane that may cause conductive hearing loss. This surgical procedure did not lead to any significant hearing loss. This approach was possible due to the large bulla structure of the common marmoset, although the lateral semicircular canal and vertical portion of the facial nerve should be carefully considered. This surgical method allows us to perform the safe and accurate administration of drugs without hearing loss, which is of great importance in obtaining pre-clinical proof of concept for translational research.


Trajectory association across multiple airborne cameras.
A camera mounted on an aerial vehicle provides an excellent means for monitoring large areas of a scene. Utilizing several such cameras on different aerial vehicles allows further flexibility, in terms of increased visual scope and in the pursuit of multiple targets. In this paper, we address the problem of associating objects across multiple airborne cameras. Since the cameras are moving and often widely separated, direct appearance-based or proximity-based constraints cannot be used. Instead, we exploit geometric constraints on the relationship between the motion of each object across cameras, to test multiple association hypotheses, without assuming any prior calibration information. Given our scene model, we propose a likelihood function for evaluating a hypothesized association between observations in multiple cameras that is geometrically motivated. Since multiple cameras exist, ensuring coherency in association is an essential requirement, e.g. that transitive closure is maintained between more than two cameras. To ensure such coherency we pose the problem of maximizing the likelihood function as a k-dimensional matching and use an approximation to find the optimal assignment of association. Using the proposed error function, canonical trajectories of each object and optimal estimates of inter-camera transformations (in a maximum likelihood sense) are computed. Finally, we show that as a result of associating objects across the cameras, a concurrent visualization of multiple aerial video streams is possible and that, under special conditions, trajectories interrupted due to occlusion or missing detections can be repaired. Results are shown on a number of real and controlled scenarios with multiple objects observed by multiple cameras, validating our qualitative models, and through simulation quantitative performance is also reported.


Dysregulation of the miR-25-IMPA2 axis promotes metastatic progression in clear cell renal cell carcinoma.
The molecular mechanism underlying clear cell renal cell carcinoma (ccRCC) metastasis remains unclear. We therefore aimed to elucidate the role of IMPA2 in ccRCC metastatic progression. Using the Cancer Genome Atlas (TCGA) database and immunohistochemistry (IHC) staining, we investigated differences in IMPA2 mRNA and protein expression, as well as their clinical relevance, in ccRCC. To investigate the function of IMPA2 in ccRCC metastasis, we performed in vitro migration and in vivo lung colony-forming assays. We further explored the effect of microRNA (miR)-25 on IMPA2 expression by performing a luciferase reporter assay. We show that ccRCC expresses relatively lower transcript levels of IMPA2 than normal kidney tissue. IMPA2 downregulation was greater in high-grade ccRCC than in low-grade ccRCC and was correlated with a poor prognosis in ccRCC patients. Importantly, we demonstrate that IMPA2 expression is inversely associated with the metastatic potential of ccRCC cells. We found that IMPA2 knockdown promotes, but overexpression suppresses, the cellular migration and lung colony-forming abilities of ccRCC cells. By using in silico and luciferase reporter assays, we found that IMPA2 expression is primarily influenced by miR-25 in ccRCC cells. Significantly, the inhibition of miR-25 function restored IMPA2 expression, thereby diminishing the metastatic potential of ccRCC cells. We conclude that miR-25-mediated IMPA2 downregulation constitutes a novel signature for cancer metastasis and poor outcomes in ccRCC. We further postulate that the therapeutic targeting of miR-25 can be useful for preventing the metastatic progression of ccRCC associated with IMPA2 downregulation. FUND: This study was supported by the Ministry of Science and Technology, Taiwan (MOST 107-2314-B-038-094, MOST 106-2314-B-038-069-MY3, MOST 105-2320-B-038-021-MY3 and MOST 107-2320-B-038-056).


Identification of the bHLH factor Math6 as a novel component of the embryonic pancreas transcriptional network.
Basic helix-loop-helix (bHLH) transcription factors play important roles in differentiation processes during embryonic development of vertebrates. In the pancreas, the atonal-related bHLH gene Neurogenin3 (Neurog3) controls endocrine cell fate specification in uncommitted progenitor cells. Therefore, it is likely that Neurog3-regulated factors will have important functions during pancreatic endocrine cell differentiation. The gene for the atonal-related bHLH factor Math6 was recognized as a potential target of Neurog3 in a genomic scale profiling during endocrine differentiation. Herein we have explored the role of Math6 during endocrine pancreas development. We demonstrate that the Math6 gene is a direct target of Neurog3 in vitro and that, during mouse development, Math6 is expressed in both endocrine and exocrine pancreatic precursor cells. We have investigated the role of Math6 in endocrine differentiation by over-expressing this factor in pancreatic duct cells. Math6 possesses intrinsic transcriptional repressor activity and, in contrast to Neurog3 it does not induce the endocrine differentiation program; however, it can modulate some of the pro-endocrine functions of Neurog3 in this system. In addition, we show that Math6 is broadly expressed in mouse embryonic tissues and its expression is induced by tissue-specific bHLH genes other than Neurog3. Furthermore, inactivation of the Math6 gene in the mouse results in early embryonic lethality demonstrating an essential role of this factor in organismal development. These data demonstrate that Math6 is a novel component of the pancreatic transcriptional network during embryonic development and suggest a potential role for Math6 as a modulator of the differentiation program initiated by the pro-endocrine factor Neurog3. Furthermore, our results demonstrate that Math6 is indispensable for early embryonic development and indicate a more widespread function for this factor in tissue-specific differentiation processes that are dependent on class II bHLH genes.


Antitumor effects of OSU-2S, a nonimmunosuppressive analogue of FTY720, in hepatocellular carcinoma.
Accumulating evidence suggests the therapeutic potential of the immunosuppressive agent FTY720 (fingolimod) in hepatocellular carcinoma (HCC). Based on our previous finding that FTY720 mediates apoptosis in HCC cells by activating reactive oxygen species (ROS)-protein kinase Cδ (PKCδ) signaling independent of effects on sphingosine-1-phosphate (S1P) receptors, we embarked on the pharmacological exploitation of FTY720 to develop a nonimmunosuppressive analogue with antitumor activity. This effort led to the development of OSU-2S, which exhibits higher potency than FTY720 in suppressing HCC cell growth through PKCδ activation. In contrast to FTY720, OSU-2S was not phosphorylated by sphingosine kinase 2 (SphK2) in vitro, and did not cause S1P1 receptor internalization in HCC cells or T lymphocyte homing in immunocompetent mice. Although devoid of S1P1 receptor activity, OSU-2S exhibited higher in vitro antiproliferative efficacy relative to FTY720 against HCC cells without cytotoxicity in normal hepatocytes. Several lines of pharmacological and molecular genetic evidence indicate that ROS-PKCδ-caspase-3 signaling underlies OSU-2S-mediated antitumor effects, and that differences in the antitumor activity between FTY720 and OSU-2S were attributable to SphK2-mediated phosphorylation of FTY720, which represents a metabolic inactivation of its antitumor activity. Finally, OSU-2S exhibited high in vivo potency in suppressing xenograft tumor growth in both ectopic and orthotopic models without overt toxicity. Using the molecular platform of FTY720, we developed OSU-2S, a novel PKCδ-targeted antitumor agent, which is devoid of S1P1 receptor activity and is highly effective in suppressing HCC tumor growth in vivo. These findings suggest that OSU-2S has clinical value in therapeutic strategies for HCC and warrants continued investigation in this regard.


Error analysis of moment-based modal wavefront sensing.
The shape of a focus-modulated point spread function (PSF) is used as a quick visual assessment tool of aberration modes in the PSF. Further analysis in terms of shape moments can permit quantifying the modal coefficients with an accuracy comparable to that of typical wavefront sensors. In this Letter, the error of the moment-based wavefront sensing is analytically described in terms of the pixelation and photon/readout noise. All components highly depend on the (unknown) PSF shape but can be estimated from the measured PSF sampled at a reasonable spatial resolution and photon count. Numerical simulations verified that the models consistently predicted the behavior of the modal estimation error of the moment-based wavefront sensing.


Thoracic Epidural analgesia versus Rectus Sheath Catheters for open midline incisions in major abdominal surgery within an enhanced recovery programme (TERSC): study protocol for a randomised controlled trial.
Thoracic epidural analgesia (TEA) is recommended for post-operative pain relief in patients undergoing major abdominal surgery via a midline incision. However, the effectiveness of TEA is variable with high failure rates reported post-operatively. Common side effects such as low blood pressure and motor block can reduce mobility and hinder recovery, and a number of rare but serious complications can also occur following their use.Rectus sheath catheters (RSC) may provide a novel alternative approach to somatic analgesia without the associated adverse effects of TEA. The aim of this study is to compare the efficacy of both techniques in terms of pain relief, patient experience, post-operative functional recovery, safety and cost-effectiveness. This is a single-centre randomised controlled non-blinded trial, which also includes a nested qualitative study. Over a two-year period, 132 patients undergoing major abdominal surgery via a midline incision will be randomised to receive either TEA or RSC for post-operative analgesia. The primary outcome measures pain scores on moving from a supine to a sitting position at 24 hours post wound closure, and the patient experience between groups evaluated through in-depth interviews. Secondary outcomes include pain scores at rest and on movement at other time points, opiate consumption, functional recovery, morbidity and cost-effectiveness. This will be the first randomised controlled trial comparing thoracic epidurals to ultrasound-guided rectus sheath catheters in adults undergoing elective midline laparotomy. The standardised care provided by an Enhanced Recovery Programme makes this a comparison between two complex pain packages and not simply two analgesic techniques, in order to ascertain if RSC is a viable alternative to TEA. Current Controlled Trials ISRCTN81223298 (16 January 2014).


Effects of some pesticides on the vital organs of juvenile rainbow trout (Oncorhynchus mykiss).
Gill, trunk kidney, spleen, and liver of rainbow trout (Oncorhynchus mykiss) were examined after exposure to different sublethal concentrations of carbosulfan (25, 50 and 200 μgL(-1)), propineb (3, 6 and 24 mgL(-1)), and benomyl (2, 5 and 20 mgL(-1)) for 14 days. Lesions were observed in gill, trunk kidney, spleen, and liver of rainbow trout exposed to either concentration of pesticides. The most important lesions were determined in the highest concentrations of pesticides. Lamellar fusion, lamellar hyperplasia, epithelial lifting, vacuolization of epithelial tissue, epithelial necrosis, hypertrophy and sloughing of epithelium were observed on fish exposed to carbosulfan, propineb and benomyl. Fish had cell necrosis, degeneration and oedemas in liver, trunk kidney and spleen. None of these lesions were seen in control fish.


Effects of body habitus on contrast-induced acute kidney injury after percutaneous coronary intervention.
Limiting the contrast volume to creatinine clearance (V/CrCl) ratio is crucial for preventing contrast-induced acute kidney injury (CI-AKI) after percutaneous coronary intervention (PCI). However, the incidence of CI-AKI and the distribution of V/CrCl ratios may vary according to patient body habitus. We aimed to identify the clinical factors predicting CI-AKI in patients with different body mass indexes (BMIs). We evaluated 8782 consecutive patients undergoing PCI and who were registered in a large Japanese database. CI-AKI was defined as an absolute serum creatinine increase of 0.3 mg/dL or a relative increase of 50%. The effect of the V/CrCl ratio relative to CI-AKI incidence was evaluated within the low- (≤25 kg/m2) and high- (>25 kg/m2) BMI groups, with a V/CrCl ratio > 3 considered to be a risk factor for CI-AKI. A V/CrCl ratio > 3 was predictive of CI-AKI, regardless of BMI (low-BMI group: odds ratio [OR], 1.77 [1.42-2.21]; P < 0.001; high-BMI group: OR, 1.67 [1.22-2.29]; P = 0.001). The relationship between BMI and CI-AKI followed a reverse J-curve relationship, although baseline renal dysfunction (creatinine clearance <60 mL/min, 46.9% vs. 21.5%) and V/CrCl ratio > 3 (37.3% vs. 20.4%) were predominant in the low-BMI group. Indeed, low BMI was a significant predictor of a V/CrCl ratio > 3 (OR per unit decrease in BMI, 1.08 [1.05-1.10]; P < 0.001). A V/CrCl ratio > 3 was strongly associated with the occurrence of CI-AKI. Importantly, we also identified a tendency for physicians to use higher V/CrCl ratios in lean patients. Thus, recognizing this trend may provide a therapeutic target for reducing the incidence of CI-AKI.


Triple-catheter technique in the transvenous coil embolization of an isolated sinus dural arteriovenous fistula.
Dural arteriovenous fistulae involving the transverse-sigmoid sinus, which is occluded at its proximal and distal ends (i.e., an isolated sinus), carry a high risk of intracranial hemorrhage or progressive neurological deficits. Although transvenous coil embolization is a useful and safe treatment for such lesions, it is often difficult to reach into the isolated sinus through the occluded sinus using the percutaneous catheter approach. We report the successful treatment of two patients with transverse-sigmoid dural arteriovenous fistulae with isolated sinus using the percutaneous transvenous triple-catheter technique. A 6-French guiding catheter was placed at the internal jugular vein followed by a second 4-French catheter positioned at the end of the occluded sinus. A third microcatheter was then navigated into the isolated sinus with support of the second catheter. Although initial attempts to reach into the isolated sinus without the second catheter failed, insertion of the second catheter resulted in successful navigation of the third microcatheter into the affected sinus in both cases. Complete cure was obtained in both cases by coil packing of the affected sinus. Although careful maneuvering is required, this triple-catheter technique is useful for treatment of dural arteriovenous fistulae with isolated sinus.


A reappraisal of Bateman's classic study of intrasexual selection.
Bateman's (1948) study showing greater variances in number of mates and reproductive success in male than female Drosophila melanogaster is a foundational paper in sexual selection. Here we show for the first time that his methods had flaws, including the elimination of genetic variance, sampling biases, miscalculations of fitness variances, statistical pseudo-replication, and selective presentation of data. We conclude that Bateman's results are unreliable, his conclusions are questionable, and his observed variances are similar to those expected under random mating. Despite our analysis, we do not intend this article as a criticism of Bateman; he accomplished his work without modern computational tools, and his approach was groundbreaking emphasizing the significance of fitness variance for sexual selection. However, this reanalysis has implications for what counts as evidence for sexual selection and we believe that our concerns should be of interest to contemporary students of sexual selection. We call for repetitions of Bateman's study using modern statistical and molecular methods.


One versus two doses: What is the best use of vaccine in an influenza pandemic?
Avian influenza A (H7N9), emerged in China in April 2013, sparking fears of a new, highly pathogenic, influenza pandemic. In addition, avian influenza A (H5N1) continues to circulate and remains a threat. Currently, influenza H7N9 vaccines are being tested to be stockpiled along with H5N1 vaccines. These vaccines require two doses, 21 days apart, for maximal protection. We developed a mathematical model to evaluate two possible strategies for allocating limited vaccine supplies: a one-dose strategy, where a larger number of people are vaccinated with a single dose, or a two-dose strategy, where half as many people are vaccinated with two doses. We prove that there is a threshold in the level of protection obtained after the first dose, below which vaccinating with two doses results in a lower illness attack rate than with the one-dose strategy; but above the threshold, the one-dose strategy would be better. For reactive vaccination, we show that the optimal use of vaccine depends on several parameters, with the most important one being the level of protection obtained after the first dose. We describe how these vaccine dosing strategies can be integrated into effective pandemic control plans.


Significance of surface charge and shell material of superparamagnetic iron oxide nanoparticle (SPION) based core/shell nanoparticles on the composition of the protein corona.
As nanoparticles (NPs) are increasingly used in many applications their safety and efficient applications in nanomedicine have become concerns. Protein coronas on nanomaterials' surfaces can influence how the cell "recognizes" nanoparticles, as well as the in vitro and in vivo NPs' behaviors. The SuperParamagnetic Iron Oxide Nanoparticle (SPION) is one of the most prominent agents because of its superparamagnetic properties, which is useful for separation applications. To mimic surface properties of different types of NPs, a core-shell SPION library was prepared by coating with different surfaces: polyvinyl alcohol polymer (PVA) (positive, neutral and negative), SiO2 (positive and negative), titanium dioxide and metal gold. The SPIONs with different surfaces were incubated at a fixed serum : nanoparticle surface ratio, magnetically trapped and washed. The tightly bound proteins were quantified and identified. The surface charge has a great impact on protein adsorption, especially on PVA and silica where proteins preferred binding to the neutral and positively charged surfaces. The importance of surface material on protein adsorption was also revealed by preferential binding on TiO2 and gold coated SPION, even negatively charged. There is no correlation between the protein net charge and the nanoparticle surface charge on protein binding, nor direct correlation between the serum proteins' concentration and the proteins detected in the coronas.


HIV-Associated Central Nervous System Disease in Patients Admitted at the Douala General Hospital between 2004 and 2009: A Retrospective Study.
Background. Studies on HIV-associated central nervous system (CNS) diseases in Cameroon are rare. The aim of this study was to describe the clinical presentation, identify aetiological factors, and determine predictors of mortality in HIV patients with CNS disease. Methods. From January 1, 2004 and December 31, 2009, we did at the Douala General Hospital a clinical case note review of 672 admitted adult (age ≥ 18 years) HIV-1 patients, and 44.6% (300/672) of whom were diagnosed and treated for HIV-associated CNS disease. Results. The mean age of the study population was 38.1 ± 13.5 years, and median CD4 count was 49 cells/mm(3) (interquartile range (QR): 17-90). The most common clinical presentations were headache (83%), focal signs (40.6%), and fever (37.7%). Toxoplasma encephalitis and cryptococcal meningitis were the leading aetiologies of HIV-associated CNS disease in 32.3% and 25% of patients, respectively. Overall mortality was 49%. Primary central nervous system lymphoma (PCNSL) and bacterial meningitis had the highest case fatality rates of 100% followed by tuberculous meningitis (79.8%). Low CD4 count was an independent predictor of fatality (AOR: 3.2, 95%CI: 2.0-5.2). Conclusions. HIV-associated CNS disease is common in Douala. CNS symptoms in HIV patients need urgent investigation because of their association with diseases of high case fatality.


Review of Geant4-DNA applications for micro and nanoscale simulations.
Emerging radiotherapy treatments including targeted particle therapy, hadron therapy or radiosensitisation of cells by high-Z nanoparticles demand the theoretical determination of radiation track structure at the nanoscale. This is essential in order to evaluate radiation damage at the cellular and DNA level. Since 2007, Geant4 offers physics models to describe particle interactions in liquid water at the nanometre level through the Geant4-DNA Package. This package currently provides a complete set of models describing the event-by-event electromagnetic interactions of particles with liquid water, as well as developments for the modelling of water radiolysis. Since its release, Geant4-DNA has been adopted as an investigational tool in kV and MV external beam radiotherapy, hadron therapies using protons and heavy ions, targeted therapies and radiobiology studies. It has been benchmarked with respect to other track structure Monte Carlo codes and, where available, against reference experimental measurements. While Geant4-DNA physics models and radiolysis modelling functionalities have already been described in detail in the literature, this review paper summarises and discusses a selection of representative papers with the aim of providing an overview of a) geometrical descriptions of biological targets down to the DNA size, and b) the full spectrum of current micro- and nano-scale applications of Geant4-DNA.


COST-effectiveness of salmeterol/fluticasone propionate combination (Advair(®)) in uncontrolled asthma in Canada.
To evaluate the cost-utility of the treatment with a long acting beta-agonist (LABA) and inhaled corticosteroid (ICS) combination inhaler [salmeterol xinafoate (SAL)/fluticasone propionate (FP) combination inhaler (SFC) (Advair(®))] to continuing on current ICS dose (no ICS dose change) or increased ICS dose [fluticasone propionate (FP)] in patients with uncontrolled asthma in Canada. A cost-utility analysis was conducted from a Canadian public healthcare perspective with a one year time horizon. In the no FP dose change scenarios, remaining on daily low (FP 100 ug BID) or medium (FP 200-250 ug BID) or high dose (FP 500 ug BID) was considered. In the increased FP dose scenarios, doubling the FP dose from low to medium dose and from medium to high dose regimens were considered. A decision model was developed with two health states: "symptom free" or "with symptoms". Clinical efficacy was based on a meta-analysis of relevant randomized controlled trials. Over the one year time horizon the percentage with symptom free days (SFD) was used as the measure of differential treatment scenario effectiveness. Drug costs and non-drug costs were incorporated into the analysis. Utilities, derived from EQ5D scores and health services resource use based on patient diaries for 'symptom free' and 'with symptoms' were based on regression analyses of individual patient data from the Gaining Optimal Asthma controL (GOAL) trial. Costs were assessed by assigning unit cost for each health services resource use for each patient. The incremental cost-utility ratios (ICUR) for SFC vs no FP dose change or increased FP dose were estimated using descriptive statistics. Uncertainty was assessed by deterministic and probabilistic sensitivity analysis (PSA). Over one year, SFC resulted in an incremental cost per patient of $544-$655 compared to no FP dose change and $47-$380 per year compared to increased FP dose. SFC results in incremental QALYs per patient of 0.0100-0.0149 compared to no FP dose change and 0.0136-0.0152 compared to increased FP dose. The one year ICURs were $43,000 to $54,400 per QALY gained for SFC compared to no FP dose change and $25,000 to $3500 per QALY gained compared to increased FP dose scenarios. The probability of SFC being cost-effective at $50,000 per QALY gained was greater than 75% compared to increased FP dose scenarios and compared to no dose change for patients on low or medium dose FP. The results were robust to changes in assumptions within the model. In Canadian patients with inadequately controlled asthma on FP, it is cost-effective to use SFC for patients 12 years and over compared to doubling their FP dose. It is also cost-effective to use SFC for patients on low or medium dose FP compared to remaining on the current FP dose in patients with uncontrolled asthma.


Design considerations for the synthesis of polymer coated iron oxide nanoparticles for stem cell labelling and tracking using MRI.
Iron oxide nanoparticles (IONPs, sometimes called superparamagnetic iron oxide nanoparticles or SPIONs) have already shown promising results for in vivo cell tracking using magnetic resonance imaging (MRI). To fully exploit the potential of these materials as contrast agents, there is still a need for a greater understanding of how they react to physiological conditions. A key aspect is the specific nature of the surface coating, which can affect important properties of the IONPs such as colloidal stability, toxicity, magnetism and labelling efficiency. Polymers are widely used as coatings for IONPs as they can increase colloidal stability in hydrophilic conditions, as well as protect the iron oxide core from degradation. In this tutorial review, we will examine the design and synthesis approaches currently being employed to produce polymer coated IONPs as cell tracking agents, and what considerations must be made. We will also give some perspective on the challenges and limitations that remain for polymer coated IONPs as MRI contrast agents for stem cell tracking.


On-demand one-step synthesis of small-sized fluorescent-magnetic bifunctional microparticles on a droplet-splitting chip.
A facile and controllable method for one-step synthesis of small-sized monodisperse fluorescent-magnetic bifunctional microparticles based on droplet splitting and photopolymerization was established. Using this method, magnetic nanoparticles and acryloyl rhodamine B (acryloyl-RhB) molecules were incorporated into the dispersed phase, which allowed the direct generation of functional microparticles. Functional microparticles with diameters in the range of 2.8 to 19.2 μm were obtained by controlling the fluidic characteristics. The fluorescence intensity of the microparticles could be regulated by adjusting the concentration of the acryloyl-RhB molecule in the dispersed phase. Furthermore, the fluorescent-magnetic bifunctional microparticles were successfully used to detect proteins specifically by ratiometric fluorescence analysis, showing their potential in biological sample detection.


Regulation of cyclin E transcription by E2Fs and retinoblastoma protein.
Cyclin E is critical for the advance of cells through the G1 phase of their growth cycle. Transcription of the cyclin E gene is known to be cell cycle-dependent. We have shown previously that mRNA levels of cyclin E are regulated positively by mitogens and negatively by TGF-beta. Much circumstantial evidence implicates both E2F transcription factors and the retinoblastoma protein (pRB) in the control of cyclin E expression. However, the molecular basis of this control has remained unclear. We report here the cloning of the cyclin E promoter and the identification of several putative E2F binding sites within the promoter sequence. We have found that cell cycle regulation of cyclin E transcription is mediated by E2F binding sites present in the promoter. The activity of this promoter can be regulated negatively by pRB. Our results suggest the operation of a positive-feedback loop in late G1 that functions to ensure continued cyclin E expression and pRB inactivation.


Static magnets for reducing pain: systematic review and meta-analysis of randomized trials.
Static magnets are marketed with claims of effectiveness for reducing pain, although evidence of scientific principles or biological mechanisms to support such claims is limited. We performed a systematic review and meta-analysis to assess the clinical evidence from randomized trials of static magnets for treating pain. Systematic literature searches were conducted from inception to March 2007 for the following data sources: MEDLINE, EMBASE, AMED (Allied and Complementary Medicine Database), CINAHL, Scopus, the Cochrane Library and the UK National Research Register. All randomized clinical trials of static magnets for treating pain from any cause were considered. Trials were included only if they involved a placebo control or a weak magnet as the control, with pain as an outcome measure. The mean change in pain, as measured on a 100-mm visual analogue scale, was defined as the primary outcome and was used to assess the difference between static magnets and placebo. Twenty-nine potentially relevant trials were identified. Nine randomized placebo-controlled trials assessing pain with a visual analogue scale were included in the main meta-analysis; analysis of these trials suggested no significant difference in pain reduction (weighted mean difference [on a 100-mm visual analogue scale] 2.1 mm, 95% confidence interval -1.8 to 5.9 mm, p = 0.29). This result was corroborated by sensitivity analyses excluding trials of acute effects and conditions other than musculoskeletal conditions. Analysis of trials that assessed pain with different scales suggested significant heterogeneity among the trials, which means that pooling these data is unreliable. The evidence does not support the use of static magnets for pain relief, and therefore magnets cannot be recommended as an effective treatment. For osteoarthritis, the evidence is insufficient to exclude a clinically important benefit, which creates an opportunity for further investigation.


Exploring the capabilities of optical pump X-ray probe NEXAFS spectroscopy to track photo-induced dynamics mediated by conical intersections.
X-ray spectroscopy is gaining a growing interest in the scientific community, as it represents a versatile and powerful experimental toolbox for probing the dynamics of both core and valence electronic excitations, nuclear motions and material structure, with element and site specificity. Among the various X-ray based techniques, near-edge X-ray absorption fine structure (NEXAFS) spectroscopy, which investigates the energy and probability of resonant core-to-valence transitions, has started to be applied to organic molecules: a recent UV-pump X-ray probe time-resolved NEXAFS experiment [Wolf et al., Nat. Commun., 2017, 8, 1] has shown the capability of the technique to provide information about the ultrafast internal conversion between the bright ππ* and the dark nπ* electronic states of the nucleobase thymine. In the present contribution we introduce an accurate theoretical approach for the simulation of NEXAFS spectra of organic molecules, employing azobenzene as a test case. The electronic structure calculations, which provide both energy levels and transition probabilities of core-to-valence excitations, were here performed with a high level multiconfigurational method, the restricted active space self consistent field (RASSCF/RASPT2). GS- and nπ*-NEXAFS spectra were obtained on the top of key molecular geometries (as the optimized cis, trans and conical intersection(s) structures) as well as along the fundamental isomerization coordinates (namely, symmetric and asymmetric bendings of the phenyl rings, and torsion around the central dihedral). We eventually characterize and explain the origin of the simulated signals, highlighting the specific signatures that make it possible to follow the excited state evolution from the nπ* Franck-Condon point, towards the conical intersection(s).


N-heterocyclic carbene tethered amido complexes of palladium and platinum.
With a view to applications in bifunctional catalysis, a modular cross-coupling strategy has been used to prepare amine bis(imidazolium) salts (3a and 3b) and an amine mono(imidazolium) salt (6) as precursors to chelating amido-NHC ligands. Treating the pro-ligands 3 with 3 equivalents of the bulky base KHMDS and Pd(OAc)(2) or PtCl(2)(COD) gave the four amido bis(N-heterocyclic carbene) pincer complexes [CNC-R]M-I [M = Pd (7) or Pt (8); R = i-Pr (a) or n-Bu (b)], including the first examples of platinum complexes of a CNC ligand. The reaction of 7a with AgOTf in pyridine gave the cationic complex {[CNC-i-Pr]Pd-py}OTf (9a). Heating a mixture of amine mono(imidazolium) salt 6 with PdCl(2) or K(2)PtCl(4), K(2)CO(3) and KI in pyridine at 100 °C gave the complexes [C,NH]MI(2)py [M = Pd (10) or Pt (11)], in which the amine arm of the NHC ligand is not deprotonated and does not coordinate to the metal. For a solution of 10 in 1,4-dioxane, deprotonation of the amine occurred in a biphasic reaction with aqueous KOH at 40 °C, giving the dimeric amido complex {[C,N]Pd(μ-OH)}(2) (12). The more inert Pt analogue 11 was unreactive under the same conditions. Solid-state structures of the complexes 7a, 7b, 9a, 10, 11 and 12 have been determined by single crystal X-ray diffraction.


Effectiveness of computerized decision support systems linked to electronic health records: a systematic review and meta-analysis.
We systematically reviewed randomized controlled trials (RCTs) assessing the effectiveness of computerized decision support systems (CDSSs) featuring rule- or algorithm-based software integrated with electronic health records (EHRs) and evidence-based knowledge. We searched MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and Cochrane Database of Abstracts of Reviews of Effects. Information on system design, capabilities, acquisition, implementation context, and effects on mortality, morbidity, and economic outcomes were extracted. Twenty-eight RCTs were included. CDSS use did not affect mortality (16 trials, 37395 patients; 2282 deaths; risk ratio [RR] = 0.96; 95% confidence interval [CI] = 0.85, 1.08; I(2) = 41%). A statistically significant effect was evident in the prevention of morbidity, any disease (9 RCTs; 13868 patients; RR = 0.82; 95% CI = 0.68, 0.99; I(2) = 64%), but selective outcome reporting or publication bias cannot be excluded. We observed differences for costs and health service utilization, although these were often small in magnitude. Across clinical settings, new generation CDSSs integrated with EHRs do not affect mortality and might moderately improve morbidity outcomes.


Limiting dilution analysis of interleukin-2-producing mature T cells. Interleukin 2 secretion is an exclusive property of L3T4+ lymphocytes.
Evaluation of lymphokine production by individual activated T cells is necessary to characterize their growth requirements. We have studied interleukin 2 (IL-2) secretion by mature T lymphocytes using a high resolution assay system with the following characteristics: (a) a threshold of IL-2 detection 25 times lower than classic IL-2 titration; (b) the ability to discriminate between IL-2 and IL-4 activities; (c) absence of 'in situ' IL-2 absorption; (d) IL-2 production revealed at the single cell level. By this method an average of 10% of spleen cells, and 75% of L3T4+ cells were detected as producers in a concanavalin A (Con A)-dependent T cell activation system. Our results also suggest the complete restriction of IL-2 secretion to cells with this phenotype. Therefore, factors other than IL-2 must play a major role in Lyt 2+ mitogen-driven, helper-independent T cell proliferation.


Hearing impairment in hypothyroid dwarf mice caused by mutations of the thyroid peroxidase gene.
Thyroid hormone (TH) is essential for proper cochlear development and function, and TH deficiencies cause variable hearing impairment in humans and mice. Thyroid peroxidase (TPO) catalyzes key reactions in TH synthesis, and TPO mutations have been found to underlie many cases of congenital hypothyroidism in human patients. In contrast, only a single mutation of the mouse TPO gene has been reported previously (Tpo(R479C)) but was not evaluated for auditory function. Here, we describe and characterize two new mouse mutations of Tpo with an emphasis on their associated auditory deficits. Mice homozygous for these recessive mutations have dysplastic thyroid glands and lack detectable levels of TH. Because of the small size of mutant mice, the mutations were named teeny (symbol Tpo(tee)) and teeny-2 Jackson (Tpo(tee-2J)). Tpo(tee) is a single base-pair missense mutation that was induced by ENU, and Tpo(tee-2J) is a 64 bp intragenic deletion that arose spontaneously. The Tpo(tee) mutation changes the codon for a highly conserved tyrosine to asparagine (p.Y614N), and the Tpo(tee-2J) mutation deletes a splice donor site, which results in exon skipping and aberrant transcripts. Mutant mice are profoundly hearing impaired with auditory brainstem response (ABR) thresholds about 60 dB above those of non-mutant controls. The maturation of cochlear structures is delayed in mutant mice and tectorial membranes are abnormally thick. To evaluate the effect of genetic background on auditory phenotype, we produced a C3.B6-Tpo(tee-2J) congenic strain and found that ABR thresholds of mutant mice on the C3H/HeJ strain background are 10-12 dB lower than those of mutant mice on the C57BL/6 J background. The Tpo mutant strains described here provide new heritable mouse models of congenital hypothyroidism that will be valuable for future studies of thyroid hormones' role in auditory development and function.


Dual beam fibre trap for Raman micro-spectroscopy of single cells.
Raman spectroscopy permits acquisition of molecular signatures from both cellular and sub-cellular samples. When combined with optical trapping we may interrogate an isolated cell reducing extraneous signals from the local environment. To date, experimental configurations have employed combinations of the single beam optical tweezers trap and Raman spectroscopy, using either the same beam or separate beams for Raman interrogation and trapping. A key problem in optical tweezers is the ability to hold and manoeuvre large cells. In this paper, we use a dual beam fibre trap to hold and manoeuvre cells combined with an orthogonally placed objective to record Raman spectra. The dual beam trap, due to its divergent light fields, offers an as yet unexploited ability to hold and move large cellular objects with reduced prospects of photodamage. We additionally show how this system permits us to move large primary human keratinocytes (approximately 30 microns in diameter), such that we may record Raman spectra from local parts of a trapped cell with ease. Finally, we develop a rudimentary microfluidic system used to generate a flow of cells. Using our dual beam trap, combined with this flow system, we hold and acquire Raman spectra from individual cells chosen from a sample of HL60 human promyelocytic leukemia cells.


Characterisation of clinically isolated Streptococcus pyogenes from balanoposthitis patients, with special emphasis on emm89 isolates.
Streptococcus pyogenes causes a variety of diseases, such as pharyngitis and toxic shock syndrome. In addition, this bacterium is a causative agent of balanoposthitis. To reveal the bacteriological characteristics of the isolates from balanoposthitis patients, we analysed 47 isolates. In addition, novel clade genotype emm89 S. pyogenes isolates have been reported to be spreading worldwide recently. Hence, we further analysed eight emm89 isolates. A drug susceptibility experiment was performed and emm types were determined. More detailed experiments, such as PCR analysis for the presence of virulence-associated genes and MLST analysis, were performed especially using emm89 isolates. All isolates were sensitive to ampicillin, but 34 % of the isolates were resistant to at least one antibiotic. The emm types of the isolates varied, with emm89 and emm11 being the most prevalent, but the emm1 type was not detected. The analysis of emm89 isolates revealed that drug susceptibilities varied. All isolates were negative for the hasABC gene and produced active NADase that are characteristics of novel clade genotype emm89 S. pyogenes. MLST analysis demonstrated that six isolates were of the ST101 type, the most predominant type reported thus far, but two isolates were of the ST646 type. According to the PCR analysis used to determine the presence of streptococcal pyrogenic exotoxin-related genes, the six ST101 isolates were further classified into four groups. These results suggest that balanoposthitis is caused by a variety of types of S. pyogenes, with novel clade genotype emm89 isolates playing a role in balanoposthitis infections in Japan.


Indices for decreased cerebral blood flow control--a modelling study.
Time-dependent interactions between pressure, flow and volume of cerebral blood and cerebrospinal fluid were mathematically modelled. The model was designed to simulate blood inflow and storage, arteriolar and capillary blood circulation controlled by cerebral autoregulation, venous blood outflow and storage modulated by intracranial pressure, and cerebrospinal fluid production, storage and reabsorption. The software implementation of the model was used to calculate the response to a gradual decrease in cerebral perfusion pressure corresponding to either systemic hypotension or intracranial hypertension. We computed flow pulsatility index (PI) and short range correlation coefficients between systolic, diastolic and mean flow velocity (FVs,d,m) and mean cerebral perfusion pressure (CPP). In simulation, the changes in cerebral flow produced by intracranial hypertension and systemic hypotension were practically indistinguishable. The relationship between PI and CPP was reciprocal, independent of the state of autoregulation. The short range running correlation coefficients between FVs, FVm and CPP indicated both combined safe CPP range and preserved autoregulation, promising a clear clinical detection of "non-worsening" blood supply conditions. A similar procedure was applied to selected clinical data to illustrate the theoretical considerations.


The associations between occupational health and safety management system programming level and prior injury and illness rates in the U.S. dairy industry.
U.S. dairy workers suffer occupational injuries and illnesses at rates higher than the national average. Occupational health and safety management systems (OHSMS) have been proposed as a way to reduce injuries and illnesses for businesses of all types and sizes. The Occupational Safety and Health Administration (OSHA) On-Site Consultation Service provides assistance establishing an OHSMS to U.S. businesses. As part of this service, the consultants determine the level of OHSMS programming using the Safety and Health Program Assessment Worksheet (Revised OSHA Form 33). A total of 167 dairy industry records were obtained from OSHA. Forty-five of those records had both injury rate and OHSMS data. Using these records, a Spearman Rank-Order correlation was used to determine the strength and significance of the associations between prior injury rates and OSHA measured OHSMS programming level for dairy operations. Additional analyses were conducted to examine potential relationships between workforce size, injury rates, and OHSMS programming levels. There was a negative correlation between OHSMS programming level and injury rates, both for the overall OHSMS and by OHSMS component. Management Leadership was the OHSMS component most strongly associated with lower injury and illness rates. OHSMS interventions, as part of a comprehensive risk management approach for the U.S. dairy industry, may be warranted to help reduce the unacceptable number of injury and illnesses in the U.S. dairy industry. Further research is needed to determine if similar relationships between OHSMS programming and injury rates occur in other industries.


Design Rationale Behind the Serious Self-Regulation Game Intervention "Balance It": Overweight Prevention Among Secondary Vocational Education Students in The Netherlands.
This article describes the design rationale behind a serious self-regulation game intervention. The aim of the game is to promote healthy dietary intake and physical activity among secondary vocational educational students in The Netherlands (approximately 16-20 years of age). The game "Balance It" was developed according to an intervention mapping (IM) protocol. Following IM, we specified health promotion and usability objectives and linked these objectives to selected behavior change and persuasive methods. Accordingly, these methods were translated into a coherent intervention program. The health behavior change objectives were derived from the determinants of the behavior and from Self-Regulation Theory (e.g., students are asked to set goals, monitor and evaluate their behavior, and create coping plans). Usability objectives were derived from the RE-AIM model (i.e., Reach, Effectiveness, Adoption, Implementation, and Maintenance). Next, behavior change and persuasive techniques were selected (e.g., goal setting and prompting, respectively) that did fit with the targeted change objectives and were translated in the design of a (mobile) serious self-regulation game intervention. Subsequently, three concept and usability tests were performed to improve intervention usability, and an evaluation plan was developed. The aim of this study was to provide a design rationale for game interventions targeting health-related behaviors. We developed a coherent program design in which both health behavior change and usability factors are addressed. The IM protocol can serve as a useful guide for decision making in program development and evaluation.


Immune-checkpoint expression in Epstein-Barr virus positive and negative plasmablastic lymphoma: a clinical and pathological study in 82 patients.
Plasmablastic lymphoma is a rare and aggressive diffuse large B-cell lymphoma commonly associated with Epstein-Barr virus co-infection that most often occurs in the context of human immunodeficiency virus infection. Therefore, its immune escape strategy may involve the upregulation of immune-checkpoint proteins allowing the tumor immune evasion. However, the expression of these molecules was poorly studied in this lymphoma. We have investigated 82 plasmablastic lymphoma cases of whom half were Epstein-Barr virus positive. Although they harbored similar pathological features, Epstein-Barr virus positive plasmablastic lymphomas showed a significant increase in MYC gene rearrangement and had a better 2-year event-free survival than Epstein-Barr virus negative cases (P=0.049). Immunostains for programmed cell death-1, programmed cell death-ligand 1, indole 2,3-dioxygenase and dendritic cell specific C-type lectin showed a high or moderate expression by the microenvironment cells in 60%-72% of cases, whereas CD163 was expressed in almost all cases. Tumor cells also expressed programmed cell death-1 and its ligand in 22.5% and 5% of cases, respectively. Both Epstein-Barr virus positive and negative plasmablastic lymphomas exhibited a high immune-checkpoint score showing that it involves several pathways of immune escape. However, Epstein-Barr virus positive lymphomas exhibited a higher expression of programmed cell death-1 and its ligand in both malignant cells and microenvironment as compared to Epstein-Barr virus negative cases. In conclusion, plasmablastic lymphoma expresses immune-checkpoint proteins through both malignant cells and the tumor microenvironment. The expression of programmed cell death-1 and its ligand constitutes a strong rationale for testing monoclonal antibodies in this often chemoresistant disease.


The impact of an integrated nursing handover system on nurses' satisfaction and work practices.
This study examined the impact of an Integrated Nursing Handover System--structured content, a minimum data set and an electronic module within the patient clinical information system--on nurses' satisfaction with handover and changes to practice. Poor transfer of patient information between clinicians at handover has been associated with adverse patient outcomes. A mixed methods pre-post evaluative approach was used. The Integrated Nursing Handover System was introduced and evaluated within an Australian hospital. Changes to nurses' satisfaction were measured using the modified Bradley Clinical Handover Survey (n = 40 pre, n = 80 post). Three focus groups with clinicians (2) and mangers and educators (1) examined changes to clinical practice. The location of handover was observed. Nurses' satisfaction with handover was improved. A two stage approach to handover emerged: nurses received handover of all patients within meeting rooms followed by handover delivered at the bedside. Major categories identified through content analysis included: implementation and the transition, work practice changes and bedside handover, accessible and standardised patient information, accountability for information transfer and a central repository of patient information. An integrated system has been implemented with positive outcomes of: improved nurse satisfaction with handover, nurses being informed about all patients, enhanced patient transfers and improved patient information for all health professionals. Further research into the potential use of stored patient handover data for research is recommended. This comprehensive system of nursing handover represents the first integrated system of this nature ever reported in the nursing and health literature. This integrated nursing handover system has been successfully implemented resulting in delivery of more comprehensive, logical and standardised patient information at handover.


Discovery and characterization of a new cell-penetrating protein.
We describe a new cell-penetrating protein, B1, capable of delivering conjugated proteins and nucleic acids into mammalian cells. B1 is a 244-amino-acid product of a single-base frameshift in the gene encoding enhanced green fluorescent protein (eGFP). The molecule has a net positive charge of 43 and a very high charge-to-mass ratio of 1.5. eGFP-fused B1 potently penetrates both adherent and suspension cells with >80% of cells taking up the protein when exposed to concentrations as low as 1 μM. The protein was found to cluster in the paranuclear region of TZM-bl cells. Most importantly, we show that B1 not only facilitates cellular uptake but allows biomolecular cargo to reach sites of biological relevance. For example, baby hamster kidney cells underwent DNA recombination when exposed to B1-tagged Cre recombinase at protein concentrations as low as 2.5 μM, indicating potent nuclear delivery of functional protein cargos. Additionally, B1 delivers noncovalently conjugated RNA and DNA across the cell membrane to cytosolic and nuclear sites accessible to the cellular translation and transcription machinery, as gauged by detection of encoded reporter functions, with efficiency comparable to commercially available cationic lipid reagents. B1 appears to utilize cell-surface glycans and multiple competing endocytic pathways to enter and traffic through cells. These studies provide both a new tool for intracellular delivery of biomolecules and insights that could aid in the design of more effective cell penetrating proteins.


Genetic counselling and consent for tumour testing in HNPCC.
Molecular pathological tests are performed on stored tumour material in order to identify individuals with hereditary non-polyposis colorectal cancer. We have previously identified that there is widespread use of this testing and now describe what counselling occurs prior to testing and the approaches in seeking consent. A respondent from every cancer genetic centre in UK offering microsatellite instability and/or immunohistochemistry testing (n= 20, response rate = 100%) was interviewed in order to ascertain pre-test counselling and consent protocols. Individuals providing consent are not always seen in person prior to providing consent but few services had supporting written information. Nine (of 19) consent forms documented consent to perform genetic testing, while the majority (14/19) sought consent to release pathology samples to the genetic service. Less than half of the services routinely seek consent to test samples from a deceased individual. Concerns were raised about spousal consent when the implications of results are for blood relatives. The differences identified between genetic counselling for testing of tumour tissue and for germ-line genetic testing suggest that counselling protocols specific for somatic testing should be developed. The results are discussed in the context of a changing legal environment and anticipated growing demand for testing.


Ending preventable child deaths in South Africa: What role can ward-based outreach teams play?
South Africa (SA) has emerged from the Millennium Development Goal era with a mixture of success and failure. The successful national scale-up of prevention of mother-to-child transmission of HIV services with increasingly efficacious antiretroviral regimens has reduced the mother-to-child transmission rate dramatically; however, over the same period there appears to have been no progress in coverage of high-impact interventions for pneumonia and diarrhoea, which are now leading causes of under-5 mortality. SA embarked on a strategy to re-engineer the primary healthcare system in 2011, which included the creation of ward-based outreach teams consisting of community health workers (CHWs). In this article we argue that the proposed ratio of CHWs to population is too low for public health impact and that the role and scope of CHWs should be extended beyond giving of health information to include assessment and treatment of childhood illnesses (particularly diarrhoea and suspected pneumonia). Evidence and experience amply demonstrate that CHWs in sufficient density can have a rapid and positive impact on neonatal and young child mortality, especially when they are allowed to treat common acute conditions. SA's mediocre performance in child survival could be dramatically improved if there were more CHWs who were allowed to do more.


Characterizing the nature of visual conscious access: the distinction between features and locations.
The difference between the roles of features and locations has been a central topic in the theoretical debates on visual attention. A recent theory proposed that momentary visual awareness is limited to one Boolean map, that is the linkage of one feature per dimension with a set of locations (L. Huang & H. Pashler, 2007). This theory predicts that: (a) access to the features of a set of objects is inefficient whereas access to their locations is efficient; (b) shuffling the locations of objects disrupts access to their features whereas shuffling the features of objects has little impact on access to their locations. Both of these predictions were confirmed in Experiments 1 and 2. Experiments 3 and 4 showed that this feature/location distinction remains when the task involves the detection of changes to old objects rather than the coding of new objects. Experiments 5 and 6 showed that, in a pre-specified set, one missing location can be readily detected, but detecting one missing color is difficult. Taken together, multiple locations seem to be accessed and represented together as a holistic pattern, but features have to be handled as separate labels, one at a time, and do not constitute a pattern in featural space.


Stereotactic Pelvic Navigation With Augmented Reality for Transanal Total Mesorectal Excision.
Technical difficulty and unfamiliar surgical anatomy are the main challenges in transanal total mesorectal excision. Precise 3-dimensional real-time image guidance may facilitate the safety, accuracy, and efficiency of transanal total mesorectal excision. A preoperative CT was obtained with 10 skin fiducials and further processed to emphasize the border of the anatomical structure by 3-dimensional modeling and pelvic organ segmentation. A forced sacral tilt by placing a 10-degree wedge under the patient's sacrum was induced to minimize pelvic organ movement caused by lithotomy position. An optical navigation system with cranial software was used. Preoperative CT images were loaded into the navigation system, and patient tracker was mounted onto the iliac bone. Once the patient-to-image paired point registration using skin fiducials was completed, the laparoscopic instrument mounted with instrument tracker was calibrated for instrument tracking. After validating the experimental setup and process of registration by navigating laparoscopic anterior resection, stereotactic navigation for transanal total mesorectal excision was performed in the low rectal neuroendocrine tumor. The fiducial registration error was 1.7 mm. The accuracy of target positioning was sufficient at less than 3 mm (1.8 ± 0.9 mm). Qualitative assessment using a Likert scale was well matched between the 2 observers. Of the 20 scores, 19 were judged as 4 (very good) or 5 (excellent). There was no statistical difference between mean Likert scales of the abdominal or transanal landmarks (4.4 ± 0.5 vs 4.3 ± 1.0, p = 0.965). Application of an existing navigation system to transanal total mesorectal excision for a low rectal tumor is feasible. The acceptable accuracy of target positioning justifies its clinical use. Further research is needed to prove the clinical need for the procedure and its impact on clinical outcomes.


Surgical Clipping of Very Small Unruptured Intracranial Aneurysms: A Multicenter International Study.
Treatment of very small unruptured intracranial aneurysms (VSUIAs, defined as ≤3 mm) can be indicated in selected circumstances. The feasibility and outcomes of endovascular therapy for VSUIAs have been recently published; however, the efficacy and complication rate of surgical clipping has not been reported in any large series to date. We conducted a multicenter study to examine surgical outcomes for VSUIAs. All consecutive patients undergoing surgery for a VSUIA in 4 neurosurgical centers between October 2001 and December 2012 were retrospectively analyzed. In the study, 183 patients (128 women, mean age 51.3 years) were treated with 190 procedures for a total of 228 aneurysms. Most were anterior circulation aneurysms (n = 215). The majority were directly clipped (n = 222, 97.4%), with coagulation or wrapping in the remainder. After 1 reoperation for incomplete clipping, postoperative imaging of 225 aneurysms confirmed complete occlusion in 221 (98.2%), 1 neck remnant (0.44%), and 3 partial occlusions (1.3%). Mortality was 0%. Early postoperative neurological deficit developed in 12 patients (6.6%); posterior circulation location was a significant risk factor for early neurological deficit (P < .001). Middle cerebral artery aneurysms had the lowest rate of postoperative deficits at 1.5% (P = .023). After the initial 30-day perioperative period, all deficits related to treatment of posterior circulation aneurysms recovered; overall neurological morbidity decreased to 2.7% with no mortality. VSUIA clipping is highly effective and is associated with a low morbidity rate. For VSUIAs selected for treatment, our data support surgical clipping as the modality of choice.


Impact of a statewide trauma system on rural emergency department patient assessment documentation. OHSU Rural Trauma Research Group.
To determine the association of rural ED patient assessment documentation with state trauma system implementation, hospital trauma categorization level (i.e., Level-3 vs Level-4), injury diagnosis, and patient demographics. A pre- vs post-system implementation (historical control) analysis of trauma documentation was performed using a sample of rural ED trauma patients from 4 Level-3 and 5 Level-4 trauma hospitals. The medical records of patients with specific index diagnoses in 4 anatomic regions (head, chest, liver/spleen, and femur/open-tibia) were reviewed for 3-year periods before statewide trauma system implementation and after hospital categorization. Vital sign, % inspired O2, and O2 saturation determinations were identified relative to the first and the last vital signs documented on the ED record. If not documented in the medical chart within 5 minutes of the first or last ED vital sign assessment, these measurements were considered missing. Separately, neurologic documentation (initial and final) also was sought for patients meeting criteria for an index head injury. Of 1,057 patients entered into the database, 532 were evaluated during the pre-system period and 525 were evaluated during the post-system period. Overall, 47% had a head injury, 34% had a chest injury, 23% had a femur/open-tibia injury, and 12% had a spleen/liver injury. There were 142 (13%) patients with an injury in > 1 index area. Except for initial systolic blood pressure, documentation of all other initial and final patient vital signs increased significantly (p < 0.05). Documentation of the Glasgow Coma Scale score (initial and final; p = 0.0001) and a final pupil examination on head-injured patients (p = 0.025) also increased. The effects of hospital level, injury diagnosis, and patient demographics on documentation rate were minimal. The study found overall improved ED documentation of trauma patient status in association with implementation of a statewide trauma system. This improvement in documentation suggests an enhanced process of care with trauma system participation.


Characterization and anti-microbial susceptibility of bacterial isolates: Experience from a tertiary care cancer center in Delhi.
Hospitalization for fever in cancer patients is associated with considerable morbidity, mortality, and cost. The aim of this study was to study the bacterial spectrum and susceptibility patterns of pathogens in culture positive patients from the oncology unit of our hospital. We retrospectively reviewed the medical records of patients admitted in our cancer center (medical, radiation, and surgical oncology) from January to December 2013. Blood and respiratory secretions from the indoor patients were evaluated. Of the total 693 samples, 76.4% were Gram-negative and 23.6% were Gram-positive. The most common bacterial isolates among Gram-negative organisms in blood were Escherichia coli, Salmonella and among the Gram-positive organism were Staphylococcus aureus and Enterococcus. Among the blood isolates extended spectrum of beta-lactamase, multidrug-resistant (carbapenem-resistant) and pan resistant bugs were seen in 47%, 15%, and 5% of the blood isolates. Among the Gram-positive organisms, 25% respiratory isolates were vancomycin-resistant Enterococci. We observed a high incidence of Gram-negative isolates with clinically significant resistance to first-line antibiotics such as cephalosporin's, piperacillin tazobactum, and fluoroquinolones.


Evolution of changes in cognitive function after the initiation of antiretroviral therapy.
Cognitive function is reported to improve after the initiation of combination antiretroviral therapy (cART). Data on the evolution of such changes are limited. We assessed the dynamics of changes in cognitive parameters, in HIV-positive subjects initiating cART. Cognitive function in seven domains was evaluated for HIV-infected patients without clinically significant cognitive impairment prior to the initiation of cART, and 24 and 48 weeks after. Cognitive scores were transformed using standardised z-scores according to the pooled baseline standard deviation. Global, speed, and accuracy composite z-scores were calculated with changes calculated using a paired t test. In 14 subjects, change in global cognitive z-scores from baseline was by 0.08 at week 24 (p = 0.59) and 0.15 at week 48 (p = 0.43). Change in composite speed and accuracy z-scores from baseline at weeks 24/48 were 0.07/0.05 (p = 0.45/0.82) and 0.13/0.23 (p = 0.47/0.45), respectively. In two of the cognitive domains assessing speed (learning and monitoring time), a continued improvement from baseline to weeks 24 and 48 was observed (changes of 0.06-0.08 and 0.10-0.19, respectively), whereas in two domains (detection and identification) an initial improvement at week 24 (changes of -0.10 and 0.04 from baseline, respectively) was followed by a deterioration in score at week 48 (changes of -0.12 and -0.08 from baseline, respectively). None of these changes were statistically significant. A trend for improvement in cognitive function was observed in naïve HIV-positive patients starting cART. The dynamics of this improvement differed both between cognitive domains and the time-points assessed.


Rational Design and Hierarchical Assembly of a Genetically Engineered Resilin-Silk Copolymer Results in Stiff Hydrogels.
Genetically engineered protein polymers, which can combine different unique peptide sequences from natural protein materials, offer great opportunities for making advanced materials with well-defined structures and properties. Here we report for the first time biosynthesis and self-assembly of a recombinant resilin-silk (RS) copolymer consisting of repeating units of silk and resilin blocks. The copolymer in aqueous solution self-assembled into nanoparticles, and the assembled nanoparticles further form nano- to microscale fibers in a time-dependent manner at body temperature, whereas such fibers were not formed upon incubation of the copolymer at either low or high temperatures. In contrast, a resilin-like polypeptide without the silk blocks exhibited a typical thermoresponsive dual-phase transition behavior and was incapable of self-assembling into fibers. More interestingly, the microscale fibers self-assembled from a moderately concentrated RS solution (20 wt %) could interact to give a self-supporting, semitransparent hydrogel with elastic modulus at approximately 195 Pa. Furthermore, photo-cross-linking of either freshly prepared or annealed RS copolymer led to the formation of stiff hydrogels and the material mechanical property was superior upon annealing of the RS solution for a longer time up to 4 h, with elastic modulus ranging from 2.9 to 7.0 kPa. These results not only shed light on the fundamental hierarchical assembly mechanism of a new family of genetically engineered RS copolymer but also suggest future opportunities for these thermoresponsive polymers in fabrication of hydrogel materials with tunable mechanical properties for diverse applications.


Two distinct mechanisms for actin capping protein regulation--steric and allosteric inhibition.
The actin capping protein (CP) tightly binds to the barbed end of actin filaments, thus playing a key role in actin-based lamellipodial dynamics. V-1 and CARMIL proteins directly bind to CP and inhibit the filament capping activity of CP. V-1 completely inhibits CP from interacting with the barbed end, whereas CARMIL proteins act on the barbed end-bound CP and facilitate its dissociation from the filament (called uncapping activity). Previous studies have revealed the striking functional differences between the two regulators. However, the molecular mechanisms describing how these proteins inhibit CP remains poorly understood. Here we present the crystal structures of CP complexed with V-1 and with peptides derived from the CP-binding motif of CARMIL proteins (CARMIL, CD2AP, and CKIP-1). V-1 directly interacts with the primary actin binding surface of CP, the C-terminal region of the alpha-subunit. Unexpectedly, the structures clearly revealed the conformational flexibility of CP, which can be attributed to a twisting movement between the two domains. CARMIL peptides in an extended conformation interact simultaneously with the two CP domains. In contrast to V-1, the peptides do not directly compete with the barbed end for the binding surface on CP. Biochemical assays revealed that the peptides suppress the interaction between CP and V-1, despite the two inhibitors not competing for the same binding site on CP. Furthermore, a computational analysis using the elastic network model indicates that the interaction of the peptides alters the intrinsic fluctuations of CP. Our results demonstrate that V-1 completely sequesters CP from the barbed end by simple steric hindrance. By contrast, CARMIL proteins allosterically inhibit CP, which appears to be a prerequisite for the uncapping activity. Our data suggest that CARMIL proteins down-regulate CP by affecting its conformational dynamics. This conceptually new mechanism of CP inhibition provides a structural basis for the regulation of the barbed end elongation in cells.


Lethal, sublethal, and combined effects of pesticides on bees: A meta-analysis and new risk assessment tools.
Multiple stressors threaten bee health, a major one being pesticides. Bees are simultaneously exposed to multiple pesticides that can cause both lethal and sublethal effects. Risk assessment and most research on bee health, however, focus on lethal individual effects. Here, we performed a systematic literature review and meta-analysis that summarizes and re-interprets the available qualitative and quantitative information on the lethal, sublethal, and combined toxicity of a comprehensive range of pesticides on bees. We provide results (1970-2019) for multiple bee species (Bombus, Osmia, Megachile, Melipona, Partamona, Scaptotrigona), although most works focused on Apis mellifera L. (78 %). Our harmonised results document the lethal toxicity of pesticides in bees (n = 377 pesticides) and the types of sublethal testing methods and related effects that cause a sublethal effect (n = 375 sublethal experiments). We identified the most common combinations of pesticides and mode of actions tested, and summarize the experimental methods, magnitude of the interactions, and robustness of available data (n = 361 experiments). We provide open access searchable, comprehensive, and integrated list of pesticides and their levels causing lethal, sublethal, and combined effects. We report major data gaps related to pesticide's sublethal (71 %) and combined (e.g., ~99 %) toxicity. We identified pesticides and mode of actions of greatest concern in terms of sublethal (chlorothalonil, pymetrozine, glyphosate; neonicotinoids) and combined (tau-fluvalinate combinations; acetylcholinesterase inhibitors and neonicotinoids) effects. Although certain pesticides have faced regulatory restrictions in specific countries (chlorothalonil, pymetrozine, neonicotinoids), most are still widely used worldwide (e.g., glyphosate). This work aims at facilitating the implementation of more comprehensive and harmonised research and risk assessments, considering sublethal and combined effects. To ensure safeguarding pollinators and the environment, we advocate for a more refined and holistic assessment that do not only focus on lethality but uses harmonised methods to test sublethal and relevant combinations.


A comprehensive analysis of the Streptococcus pyogenes and human plasma protein interaction network.
Streptococcus pyogenes is a major human bacterial pathogen responsible for severe and invasive disease associated with high mortality rates. The bacterium interacts with several human blood plasma proteins and clarifying these interactions and their biological consequences will help to explain the progression from mild to severe infections. In this study, we used a combination of mass spectrometry (MS) based techniques to comprehensively quantify the components of the S. pyogenes-plasma protein interaction network. From an initial list of 181 interacting human plasma proteins defined using liquid chromatography (LC)-MS/MS analysis we further subdivided the interacting protein list using selected reaction monitoring (SRM) depending on the level of enrichment and protein concentration on the bacterial surface. The combination of MS methods revealed several previously characterized interactions between the S. pyogenes surface and human plasma along with many more, so far uncharacterised, possible plasma protein interactions with S. pyogenes. In follow-up experiments, the combination of MS techniques was applied to study differences in protein binding to a S. pyogenes wild type strain and an isogenic mutant lacking several important virulence factors, and a unique pair of invasive and non-invasive S. pyogenes isolates from the same patient. Comparing the plasma protein-binding properties of the wild type and the mutant and the invasive and non-invasive S. pyogenes bacteria revealed considerable differences, underlining the significance of these protein interactions. The results also demonstrate the power of the developed mass spectrometry method to investigate host-microbial relationships with a large proteomics depth and high quantitative accuracy.


Economic analysis at the global level: a resource requirement model for HIV prevention in developing countries.
Agencies operating at the international level have a need for economic analysis to help develop global health policies and determine resource requirements to support their advocacy efforts. This paper presents work commissioned by the Global Programme on AIDS to estimate the total resource requirements of implementing a package of HIV prevention strategies in developing countries. The modelling approach identified a hypothetical package which should be implemented and developed a set of assumptions relating the size, number and coverage of programmes required for each strategy to a set of demographic and other characteristics of individual countries. Costs were attached to estimate the total costs of the package for individual countries, regions and the developing world. Results are presented for regions and their implications discussed. Conclusions are drawn on the value of this type of modelling approach to estimating resource requirements.


Effect of genetic polymorphisms on effectiveness of low-dose azathioprine in Japanese patients with systemic lupus erythematosus.
The effect of genetic polymorphisms on the effectiveness of low-dose azathioprine in Japanese patients with systemic lupus erythematosus (SLE) was studied. Sixty-one unrelated Japanese patients with SLE treated with azathioprine were included in the study. The selected genetic polymorphisms evaluated in the analysis were ITPA 138G>A, ITPA 563G>A, IMPDH 1575A>G, XO 837C>T, XO 2211C>T, XO 3030C>T, and MRP4 2269G>A. The DNA isolation and genotyping procedures for ITPA 94C>A and TPMT*3C were the same as those in a previous study. Genotyping of ITPA 138G>A, ITPA 563G>A, IMPDH 1575A>G, MRP4 2269G>A, XO 837C>T, XO 2211C>T, and XO 3030C>T was performed using a validated genotyping assay. The SLE disease activity index (SLEDAI) score was used as a marker for the efficacy of azathioprine treatment, and the correlations between the changes in the SLEDAI score and the considered polymorphisms were evaluated. The mean SLE duration, SLEDAI score, and azathioprine dosage prescribed were 5.6 years, 6.0, and 1 mg/kg/day, respectively. Small (but not significant) p values suggested a tendency for the reduction in the SLEDAI score to be greater in patients with the ITPA 94A allele. Further, the ITPA 94C>A polymorphism correlated highly with the change in the SLEDAI score. However, there were no significant associations among TPMT*3C, ITPA 138G>A, ITPA 563G>A, IMPDH 1575A>G, XO 837C>T, XO 2211C>T, and XO 3030C>T and the mean changes in the SLEDAI score over the one-year investigation period. The ITPA 94C>A polymorphism was found to possibly influence the clinical response to low-dose azathioprine in Japanese patients with SLE. The other studied single-nucleotide polymorphisms appeared to have little influence on the effectiveness of azathioprine.


Escherichia coli DNA polymerase II can efficiently bypass 3,N(4)-ethenocytosine lesions in vitro and in vivo.
Escherichia coli DNA polymerase II (pol-II) is a highly conserved protein that appears to have a role in replication restart, as well as in translesion synthesis across specific DNA adducts under some conditions. Here, we have investigated the effects of elevated expression of pol-II (without concomitant SOS induction) on translesion DNA synthesis and mutagenesis at 3,N(4)-ethenocytosine (varepsilonC), a highly mutagenic DNA lesion induced by oxidative stress as well as by exposure to industrial chemicals such as vinyl chloride. In normal cells, survival of transfected M13 single-stranded DNA bearing a single varepsilonC residue (varepsilonC-ssDNA) is about 20% of that of control DNA, with about 5% of the progeny phage bearing a mutation at the lesion site. Most mutations are C-->A and C-->T, with a slight predominance of transversions over transitions. In contrast, in cells expressing elevated levels of pol-II, survival of varepsilonC-ssDNA is close to 100%, with a concomitant mutation frequency of almost 99% suggesting highly efficient translesion DNA synthesis. Furthermore, an overwhelming majority of mutations at varepsilonC are C-->T transitions. Purified pol-II efficiently catalyzes translesion synthesis at varepsilonC in vitro, accompanied by high levels of mutagenesis with the same specificity. These results suggest that the observed in vivo effects in pol-II over-expressing cells are due to pol-II-mediated DNA synthesis. Introduction of mutations in the carboxy terminus region (beta interaction domain) of polB eliminates in vivo translesion synthesis at varepsilonC, suggesting that the ability of pol-II to compete with pol-III requires interaction with the beta processivity subunit of pol-III. Thus, pol-II can compete with pol-III for translesion synthesis.


Aluminium in brain tissue in autism.
Autism spectrum disorder is a neurodevelopmental disorder of unknown aetiology. It is suggested to involve both genetic susceptibility and environmental factors including in the latter environmental toxins. Human exposure to the environmental toxin aluminium has been linked, if tentatively, to autism spectrum disorder. Herein we have used transversely heated graphite furnace atomic absorption spectrometry to measure, for the first time, the aluminium content of brain tissue from donors with a diagnosis of autism. We have also used an aluminium-selective fluor to identify aluminium in brain tissue using fluorescence microscopy. The aluminium content of brain tissue in autism was consistently high. The mean (standard deviation) aluminium content across all 5 individuals for each lobe were 3.82(5.42), 2.30(2.00), 2.79(4.05) and 3.82(5.17) μg/g dry wt. for the occipital, frontal, temporal and parietal lobes respectively. These are some of the highest values for aluminium in human brain tissue yet recorded and one has to question why, for example, the aluminium content of the occipital lobe of a 15year old boy would be 8.74 (11.59) μg/g dry wt.? Aluminium-selective fluorescence microscopy was used to identify aluminium in brain tissue in 10 donors. While aluminium was imaged associated with neurones it appeared to be present intracellularly in microglia-like cells and other inflammatory non-neuronal cells in the meninges, vasculature, grey and white matter. The pre-eminence of intracellular aluminium associated with non-neuronal cells was a standout observation in autism brain tissue and may offer clues as to both the origin of the brain aluminium as well as a putative role in autism spectrum disorder.


Non-adaptive origins of interactome complexity.
The boundaries between prokaryotes, unicellular eukaryotes and multicellular eukaryotes are accompanied by orders-of-magnitude reductions in effective population size, with concurrent amplifications of the effects of random genetic drift and mutation. The resultant decline in the efficiency of selection seems to be sufficient to influence a wide range of attributes at the genomic level in a non-adaptive manner. A key remaining question concerns the extent to which variation in the power of random genetic drift is capable of influencing phylogenetic diversity at the subcellular and cellular levels. Should this be the case, population size would have to be considered as a potential determinant of the mechanistic pathways underlying long-term phenotypic evolution. Here we demonstrate a phylogenetically broad inverse relation between the power of drift and the structural integrity of protein subunits. This leads to the hypothesis that the accumulation of mildly deleterious mutations in populations of small size induces secondary selection for protein-protein interactions that stabilize key gene functions. By this means, the complex protein architectures and interactions essential to the genesis of phenotypic diversity may initially emerge by non-adaptive mechanisms.


The use of accelerometer-based navigation for coronal TKA alignment: a prospective, single surgeon comparative study.
There is the need for a device that can be used to accurately position components during total knee arthroplasty (TKA) with minimal impact on procedure time, workflow and cost. This study aimed to investigate the accuracy and time efficiency of a novel, accelerometer-based navigation system (ABN). This prospective, single surgeon study of patients undergoing TKA for osteoarthritis over a 5 year period involved a total of 138 patients: 110 using the ABN system and 28 without. The ABN system consists of two coupled inertial pods that are secured to resection guides, providing a body-fixed 3D coordinate system for limb segments. Post-operative coronal alignment was measured from standardised long-leg AP radiographs. Deviation of the femur and tibia from the neutral coronal mechanical axis was recorded. Intra-observer repeatability was performed on three independent blinded data sets. The BMI and the surgical time (skin to skin) were recorded for all patients. The mean BMI was 34 in the ABN group and 33 in the control group (p = 0.92). The skin-to-skin time was also similar between the groups; 105 min in the navigation group and 100 min in the control group (p = 0.297). The use of navigation resulted in significantly fewer outliers as defined by < 3º deviation from the target angle. 3 of 110 navigated patients recorded an AP femur angle of more than 3º from the target of 90º, where 5 of 28 control patients fell outside of the ± 3º window (p = 0.009, Fig. 1). The use of the ABN system significantly improved accuracy of implant position and alignment without increasing surgical time.


[Critical ischemia of the diabetic patient].
Critical ischemia of the lower limbs is the final result of diabetic arteriopathy. The surgeon is more and more forced to choose between amputation d emblée and the attempt to revascularization in very old patients and in deficient general conditions. Shall we point out some principle to address our strategy? To answer this question we reviewed retrospectively our surgical choices and the results obtained during two years, from January 1997 to December 1998 (medium follow-up 11.3 months), at the Emergency Division of the Cannizzaro Hospital of Catania. In this period 143 diabetic patients were selected by our ambulatory. Of these, nineteen had a critical lower limb ischemia and therefore were submitted to a revascularization and/or amputation. All the revascularized patients were controlled by Doppler-sonography immediately after operation and then daily, till their discharge. Controls were done at the first, third, sixth month and after one year, except for patients who complained a new objective or subjective ischemic symptomatology. The primary patency rate was 84.2% and the secondary patency rate was 89.4%. The amputation rate due to the procedure failure was 5%. The complications were three: two graft infections (10.5%) and one early thrombosis of a femoro-popliteal bypass graft, due to technical defect (5.2%). Perioperative mortality rate was 15.7%. These results, if related with those reported in the letterature about amputations, are in favour of the attempt to revascularization.


Association of Jumping Ability and Maximum Strength With Dive Distance in Swimmers.
The aim of the current study was to investigate the relationship between dive distance (DD) and countermovement jump (CMJ) height, track start CMJ height, countermovement broad jump (CMBJ) distance, track start broad jump distance, and isometric midthigh pull peak force and relative peak force. A total of 27 (11 female and 16 male) regional-national-international-standard swimmers (mean [SD]; age = 19.5 [5.5] y; mass = 69.3 [10.5] kg; height = 1.77 [0.09] m) performed 3 trials of a track start dive, CMJ, track start CMJ, CMBJ, track start broad jump, and isometric midthigh pull. Data were separated into pooled (females and males combined), females, and males. Large to very large correlations were found between DD and all variables tested for pooled data (r = .554-.853, P < .001-.008), with DD-CMBJ displaying the highest correlation (r = .853, P < .001). CMBJ accounted for 70% of the variance in DD. Females demonstrated moderate nonsignificant correlations between DD isometric midthigh pull (r = .379, P < .125). Males demonstrated very large significant correlations between DD-CMJ (r = .761, P < .001). DD demonstrated strong correlations with jump performances and multijoint isometric force production in pooled data. Males showed stronger correlations than females due to being stronger and being able to perform the jumping/strength tasks to a higher standard. Enhanced jump performance and increased maximal force production may, therefore, enhance DD in swimmers.


Topographic dissociation between mitochondrial dysfunction and cell death during low-flow hypoxia in perfused rat liver.
Although it has been postulated that mitochondrial dysfunction and oxygen radicals may play a role in the mechanisms of hypoxic liver injury, little is known about the intralobular topographic relationship between a disorder in oxygen metabolism and cell death during hypoxia. Intralobular heterogeneity of mitochondrial dysfunction and its topographic relationship to cell death during 25% low-flow hypoxia was visually demonstrated in perfused rat hepatic microcirculatory units by dual-color digital microfluorography using rhodamine 123, a fluorochrome sensitive to the mitochondrial membrane potential, and propidium iodide, which labels the nuclei of nonviable cells with a concomitant enhancement of fluorescence. In the normoperfused liver from fasted rats, no significant depletion of the mitochondrial membrane potential values was observed in any portion of lobules during 100 minutes. In the liver treated with the 40-minute hypoxia, the mitochondrial membrane potential values within the entire lobule were significantly decreased. After a longer period of hypoxia, the pericentral fluorescence decreased to background levels, and the intralobular gradient showed a steep decline midway between the periportal and pericentral regions. Cell death was at first prominent in the midzone at 40 minutes, and then extended towards the pericentral regions, forming centrilobular necrosis at 100 minutes. Although allopurinol diminished the early midzonal cell death and retarded the development of centrilobular necrosis, it did not attenuate depression of the mitochondrial membrane potential. It is conceivable that mitochondrial dysfunction per se may not be related to the intralobular distribution of cell death during hypoxia. On the other hand, mitochondrial dysfunction in the centrilobular regions seems to be an important event leading to early midzonal oxidative tissue damage in that this change may involve ATP breakdown and the subsequent enhancement of xanthine oxidase-mediated oxidative cell injury in the midzonal regions, which are incompletely oxygenated.


Gene mutations versus clinically relevant phenotypes: lyso-Gb3 defines Fabry disease.
Currently, no method is available to identify α-galactosidase A (agalA) mutations determining clinically relevant Fabry disease. In our largest European Fabry cohort, we investigated whether a biomarker, specific for the defect, could stratify persons at risk. A total of 124 individuals with agalA mutations were investigated with a comprehensive clinical workup, genetic analysis, and laboratory testing, including measurements of agalA activity and lyso-Gb3 (degradation product of the accumulating Gb3). Additionally, an extensive family screening with a clinical workup of relatives was performed. The patient population was divided into 2 samples: previously described mutations (n=72) and novel mutations (n=52). The patients with previously described mutations were subdivided into 2 groups: classical mutations, which were known to cause the classic type of Fabry disease with specific symptoms and a high risk for major events in all 3 main organs (heart, kidney, and central nervous system), and atypical mutations without the typical presentation. All patients with atypical mutations (n=17) had lower lyso-Gb3 levels than any of the patients with classical Fabry disease (n=55). A cutoff value of 2.7 ng/mL separated the 2 groups. Six out of 52 patients with novel mutations showed a lyso-Gb3 level <2.7 ng/mL. Clinical investigation, blinded to lyso-Gb3 results, revealed no classic organ involvement in these patients or their relatives. In contrast, the characterization of patients with lyso-Gb3≥2.7 ng/mL suggested classical Fabry mutations in most of the patients (93%). Our data show that the biomarker lyso-Gb3 may identify the clinically relevant agalA mutations leading to Fabry disease.


Cyclopenta[b]naphthalene cyanoacrylate dyes: synthesis and evaluation as fluorescent molecular rotors.
We describe the design, synthesis and fluorescent profile of a family of environment-sensitive dyes in which a dimethylamino (donor) group is conjugated to a cyanoacrylate (acceptor) unit via a cyclopenta[b]naphthalene ring system. This assembly satisfies the typical D-π-A motif of a fluorescent molecular rotor and exhibits solvatochromic and viscosity-sensitive fluorescence emission. The central naphthalene ring system of these dyes was synthesized via a novel intramolecular dehydrogenative dehydro-Diels-Alder (IDDDA) reaction that permits incorporation of the donor and acceptor groups in variable positions around the aromatic core. A bathochromic shift of excitation and emission peaks was observed with increasing solvent polarity but the dyes exhibited a complex emission pattern with a second red emission band when dissolved in nonpolar solvents. Consistent with other known molecular rotors, the emission intensity increased with increasing viscosity. Interestingly, closer spatial proximity between the donor and the acceptor groups led to decreased viscosity sensitivity combined with an increased quantum yield. This observation indicates that structural hindrance of intramolecular rotation dominates when the donor and acceptor groups are in close proximity. The examined compounds give insight into how excited state intramolecular rotation can be influenced by both the solvent and the chemical structure.