Fast single-molecule FRET spectroscopy: theory and experiment.
Single-molecule spectroscopy is widely used to study macromolecular dynamics. Although this technique provides unique information that cannot be obtained at the ensemble level, the possibility of studying fast molecular dynamics is limited by the number of photons detected per unit time (photon count rate), which is proportional to the illumination intensity. However, simply increasing the illumination intensity often does not help because of various photophysical and photochemical problems. In this Perspective, we show how to improve the dynamic range of single-molecule fluorescence spectroscopy at a given photon count rate by considering each and every photon and using a maximum likelihood method. For a photon trajectory with recorded photon colors and inter-photon times, the parameters of a model describing molecular dynamics are obtained by maximizing the appropriate likelihood function. We discuss various likelihood functions, their applicability, and the accuracy of the extracted parameters. The maximum likelihood method has been applied to analyze the experiments on fast two-state protein folding and to measure transition path times. Utilizing other information such as fluorescence lifetimes is discussed in the framework of two-dimensional FRET efficiency-lifetime histograms.


Immune tolerance split between hepatitis B virus precore and core proteins.
The function of the hepatitis B virus (HBV) precore or HBeAg is largely unknown because it is not required for viral assembly, infection, or replication. However, the HBeAg does appear to play a role in viral persistence. It has been suggested that the HBeAg may promote HBV chronicity by functioning as an immunoregulatory protein. As a model of chronic HBeAg exposure and to examine the tolerogenic potential of the HBV precore and core (HBcAg) proteins, HBc/HBeAg-transgenic (Tg) mice crossed with T cell receptor (TCR)-Tg mice expressing receptors for the HBc/HBeAgs (i.e., TCR-antigen double-Tg pairs) were produced. This study revealed three phenotypes of HBe/HBcAg-specific T-cell tolerance: (i) profound T-cell tolerance most likely mediated by clonal deletion, (ii) T-cell clonal ignorance, and (iii) nondeletional T-cell tolerance mediated by clonal anergy and dependent on the structure, location, and concentration of the tolerogen. The secreted HBeAg is significantly more efficient than the intracellular HBcAg at eliciting T-cell tolerance. The split T-cell tolerance between the HBeAg and the HBcAg and the clonal heterogeneity of HBc/HBeAg-specific T-cell tolerance may have significant implications for natural HBV infection and especially for precore-negative chronic hepatitis.


Artificial Avidin-Based Receptors for a Panel of Small Molecules.
Proteins with high specificity, affinity, and stability are needed for biomolecular recognition in a plethora of applications. Antibodies are powerful affinity tools, but they may also suffer from limitations such as low stability and high production costs. Avidin and streptavidin provide a promising scaffold for protein engineering, and due to their ultratight binding to D-biotin they are widely used in various biotechnological and biomedical applications. In this study, we demonstrate that the avidin scaffold is suitable for use as a novel receptor for several biologically active small molecules: Artificial, chicken avidin-based proteins, antidins, were generated using a directed evolution method for progesterone, hydrocortisone, testosterone, cholic acid, ketoprofen, and folic acid, all with micromolar to nanomolar affinity and significantly reduced biotin-binding affinity. We also describe the crystal structure of an antidin, sbAvd-2(I117Y), a steroid-binding avidin, which proves that the avidin scaffold can tolerate significant modifications without losing its characteristic tetrameric beta-barrel structure, helping us to further design avidin-based small molecule receptors.


ADP-ribosylation factor/COPI-dependent events at the endoplasmic reticulum-Golgi interface are regulated by the guanine nucleotide exchange factor GBF1.
ADP-ribosylation factor (ARF) mediated recruitment of COPI to membranes plays a central role in transport between the endoplasmic reticulum (ER) and the Golgi. The activation of ARFs is mediated by guanine nucleotide exchange factors (GEFs). Although several ARF-GEFs have been identified, the transport steps in which they function are still poorly understood. Here we report that GBF1, a member of the Sec7-domain family of GEFs, is responsible for the regulation of COPI-mediated events at the ER-Golgi interface. We show that GBF1 is essential for the formation, differentiation, and translocation of pre-Golgi intermediates and for the maintenance of Golgi integrity. We also show that the formation of transport-competent ER-to-Golgi intermediates proceeds in two stages: first, a COPI-independent event leads to the formation of an unstable compartment, which is rapidly reabsorbed in the absence of GBF1 activity. Second, the association of GBF1 with this compartment allows COPI recruitment and leads to its maturation into transport intermediates. The recruitment of GBF1 to this compartment is specifically inhibited by brefeldin A. Our findings imply that the continuous recruitment of GBF1 to spatially differentiated membrane domains is required for sustained membrane remodeling that underlies membrane traffic and Golgi biogenesis.


[Alternative splicing of vascular endothelial growth factor A and ocular neovascularization].
Ocular neovascularization is the primary cause of blindness in a wide range of ocular diseases. The vascular endothelial growth factor A (VEGF-A) is the key factor involved in ocular angiogenesis, which can cause eye diseases through the development of pathological angiogenesis and increase of vascular permeability. There are two families of VEGF-A isoforms formed by alternative splicing, the angiogenic VEGF-A family (VEGF(xxx)), known to contribute to ocular neovascularization, and the anti-angiogenic VEGF-A family (VEGF(xxx)b), which is found in normal ocular tissues but downregulated in human diabetic retinopathy. The first member of the VEGF(xxx)b family to be isolated was VEGF(165)b. It can significantly reduce preretinal neovascularization without inhibition of physiological intraretinal angiogenesis. As the studies on the VEGF(xxx)b family proceed more deeply, controlling the balance of VEGF(xxx) to VEGF(xxx)b isoforms may be therapeutically valuable in the treatment of angiogenic eye diseases such as diabetic retinopathy and age-related macular degeneration.


Validation of selected analytical methods using accuracy profiles to assess the impact of a Tobacco Heating System on indoor air quality.
Studies in environmentally controlled rooms have been used over the years to assess the impact of environmental tobacco smoke on indoor air quality. As new tobacco products are developed, it is important to determine their impact on air quality when used indoors. Before such an assessment can take place it is essential that the analytical methods used to assess indoor air quality are validated and shown to be fit for their intended purpose. Consequently, for this assessment, an environmentally controlled room was built and seven analytical methods, representing eighteen analytes, were validated. The validations were carried out with smoking machines using a matrix-based approach applying the accuracy profile procedure. The performances of the methods were compared for all three matrices under investigation: background air samples, the environmental aerosol of Tobacco Heating System THS 2.2, a heat-not-burn tobacco product developed by Philip Morris International, and the environmental tobacco smoke of a cigarette. The environmental aerosol generated by the THS 2.2 device did not have any appreciable impact on the performances of the methods. The comparison between the background and THS 2.2 environmental aerosol samples generated by smoking machines showed that only five compounds were higher when THS 2.2 was used in the environmentally controlled room. Regarding environmental tobacco smoke from cigarettes, the yields of all analytes were clearly above those obtained with the other two air sample types.


Integration column: microwell arrays for mammalian cell culture.
Microwell arrays have emerged as robust and versatile alternatives to conventional mammalian cell culture substrates. Using standard microfabrication processes, biomaterials surfaces can be topographically patterned to comprise high-density arrays of micron-sized cavities with desirable geometry. Hundreds to thousands of individual cells or cell colonies with controlled size and shape can be trapped in these cavities by simple gravitational sedimentation. Efficient long-term cell confinement allows for parallel analyses and manipulation of cell fate during in vitro culture. These live-cell arrays have already found applications in cell biology, for example to probe the effect of cell colony size on embryonic stem cell differentiation, to dissect the heterogeneity in single cell proliferation kinetics of neural or hematopoietic stem/progenitor cell populations, or to elucidate the role of cell shape on cell function. Here, we highlight the key applications of these platforms, hopefully inspiring biologists to apply these systems for their own studies.


A self-tunable Titanium Sapphire laser by rotating a set of parallel plates of active material.
In a recent work, the authors reported the experimental demonstration of wavelength tuning in a single birefringent plate of Ti:sapphire crystal based on its own birefringence properties. In that device, the thickness of the active plate, limited by the width of the single order tuning spectral region, imposed a strong constraint in the power performance of the laser. The aim of this work is to overcome this limitation by using a set of several identical birefringent plates so that the wavelength tuning of the laser is obtained by synchronously rotating the plates in their own plane. A discussion about the laser performance is presented.


Intimal ultrastructure of human umbilical arteries. Observations on arteries from newborn children of smoking and nonsmoking mothers.
The umbilical artery was chosen as a possible model for evaluating the vascular injury provoked by tobacco smoking in humans. Cords from newborn children delivered by 15 nonsmoking and 13 smoking mothers were studied in the transmission and the scanning electron microscope. Pronounced intimal changes were seen in the arteries from smoking mothers; the most important findings were degenerative changes of the endothelium such as swelling, blebbing, contraction, and subsequent opening of the endothelial junctions with formation of subendothelial edema. Other observations included dilation of the endoplasmic reticulum in the endothelium and reparative changes such as a considerable widening of the basement membrane. Since similar changes can be induced in arteries of animals by exposure to carbon monoxide or perfusion with nicotine, we conclude that the present study supports the concept that tobacco smoking is harmful to the vascular endothelium. This study also contributes to an understanding of the mechanism through which vascular injury is provoked in heavy smokers.


Rationing health care: the choice before us.
Rapid technological advances and upward pressure on wages of hospital personnel are leading to a steady increase in health care spending that is absorbing an ever-larger fraction of gross national product. Eliminating inefficiencies in the system can provide brief fiscal relief, but rationing of beneficial services, even to the well-insured, offers the only prospect for sustained reduction in the growth of health care spending. The United States, which has negligible direct experience with rationing, can learn about choices it will face from the experience of Great Britain where health care has been rationed explicitly for many years.


Ponatinib inhibits polyclonal drug-resistant KIT oncoproteins and shows therapeutic potential in heavily pretreated gastrointestinal stromal tumor (GIST) patients.
KIT is the major oncogenic driver of gastrointestinal stromal tumors (GIST). Imatinib, sunitinib, and regorafenib are approved therapies; however, efficacy is often limited by the acquisition of polyclonal secondary resistance mutations in KIT, with those located in the activation (A) loop (exons 17/18) being particularly problematic. Here, we explore the KIT-inhibitory activity of ponatinib in preclinical models and describe initial characterization of its activity in patients with GIST. The cellular and in vivo activities of ponatinib, imatinib, sunitinib, and regorafenib against mutant KIT were evaluated using an accelerated mutagenesis assay and a panel of engineered and GIST-derived cell lines. The ponatinib-KIT costructure was also determined. The clinical activity of ponatinib was examined in three patients with GIST previously treated with all three FDA-approved agents. In engineered and GIST-derived cell lines, ponatinib potently inhibited KIT exon 11 primary mutants and a range of secondary mutants, including those within the A-loop. Ponatinib also induced regression in engineered and GIST-derived tumor models containing these secondary mutations. In a mutagenesis screen, 40 nmol/L ponatinib was sufficient to suppress outgrowth of all secondary mutants except V654A, which was suppressed at 80 nmol/L. This inhibitory profile could be rationalized on the basis of structural analyses. Ponatinib (30 mg daily) displayed encouraging clinical activity in two of three patients with GIST. Ponatinib possesses potent activity against most major clinically relevant KIT mutants and has demonstrated preliminary evidence of activity in patients with refractory GIST. These data strongly support further evaluation of ponatinib in patients with GIST.


Effectiveness and safety of simultaneous hybrid thoracoscopic and endocardial catheter ablation of lone atrial fibrillation.
We evaluated the safety and effectiveness of the hybrid thoracoscopic endocardial epicardial technique for the treatment of lone atrial fibrillation. Between 2009 and 2012, a cohort of 78 consecutive patients (median age 60.5 years, 77% male) underwent ablation of atrial fibrillation (AF) as a stand-alone procedure using a thoracoscopic, hybrid epicardial-endocardial technique. All patients underwent continuous 7-day Holter monitoring at 3 months, 6 months, 1 year and yearly thereafter. All patients reached 1-year follow-up. Median follow-up was 24 months [interquartile range 12-36]. No death or conversion to cardiopulmonary bypass occurred. No patient demonstrated paralysis of the phrenic nerve. Overall, the incidence of perioperative complications was 8% (n=6). At the end of follow-up, sixty-eight patients (87%) were in sinus rhythm (SR) with no episode of AF, atrial flutter or atrial tachycardia lasting longer than 30 seconds and off antiarrhythmic drugs (ADD). Among patients with long-standing persistent AF, 15 (100%) were in SR and off AAD. Success rates were 82% (n=28) in persistent and 76% (n=22) in paroxysmal AF (P=0.08). No patient died and no thromboembolic/bleeding events or procedure-related complications occurred during the follow-up. Thoracoscopic hybrid epicardial endocardial technique proved to be effective and safe in the treatment of LAF and to represent an important new suitable option to treat stand-alone AF. Our findings need to be confirmed by further larger studies.


Enhanced perception of various linguistic features by musicians: a cross-linguistic study.
Two cross-linguistic experiments comparing musicians and non-musicians were performed in order to examine whether musicians have enhanced perception of specific acoustical features of speech in a second language (L2). These discrimination and identification experiments examined the perception of various speech features; namely, the timing and quality of Japanese consonants, and the quality of Dutch vowels. We found that musical experience was more strongly associated with discrimination performance rather than identification performance. The enhanced perception was observed not only with respect to L2, but also L1. It was most pronounced when tested with Japanese consonant timing. These findings suggest the following: 1) musicians exhibit enhanced early acoustical analysis of speech, 2) musical training does not equally enhance the perception of all acoustic features automatically, and 3) musicians may enjoy an advantage in the perception of acoustical features that are important in both language and music, such as pitch and timing.


Study of larvivorous fish for malaria vector control in Somalia, 2002.
An intervention study was conducted on the introduction of the larvivorous fish Oreochromis spilurus spilurus as a method of malaria vector control in Kalabeydh village, northern Somalia. This species is resistant to chlorine in water up to a concentration of 1.0 mg/L. Fish were introduced into 25 berkit (reservoirs). After 1 month the number of larvae in each berkit was reduced by between 16.5% and 78.6% (mean 52.8%). Community acceptance and participation was good. The introduction of larvivorous fish is a cheap method of malaria vector control, but its sustainability needs special consideration and education of the community is important, especially to cover the dry season, when most of the berkit dry up.


Genetic and molecular identification of three human TPP1 functions in telomerase action: recruitment, activation, and homeostasis set point regulation.
Telomere length homeostasis is essential for the long-term survival of stem cells, and its set point determines the proliferative capacity of differentiated cell lineages by restricting the reservoir of telomeric repeats. Knockdown and overexpression studies in human tumor cells showed that the shelterin subunit TPP1 recruits telomerase to telomeres through a region termed the TEL patch. However, these studies do not resolve whether the TPP1 TEL patch is the only mechanism for telomerase recruitment and whether telomerase regulation studied in tumor cells is representative of nontransformed cells such as stem cells. Using genome engineering of human embryonic stem cells, which have physiological telomere length homeostasis, we establish that the TPP1 TEL patch is genetically essential for telomere elongation and thus long-term cell viability. Furthermore, genetic bypass, protein fusion, and intragenic complementation assays define two distinct additional mechanisms of TPP1 involvement in telomerase action at telomeres. We demonstrate that TPP1 provides an essential step of telomerase activation as well as feedback regulation of telomerase by telomere length, which is necessary to determine the appropriate telomere length set point in human embryonic stem cells. These studies reveal and resolve multiple TPP1 roles in telomere elongation and stem cell telomere length homeostasis.


Agreement of dermatopathologists in the evaluation of clinically difficult melanocytic lesions: how golden is the 'gold standard'?
The 'gold standard' for the diagnosis of melanocytic lesions is dermatopathology. Although most of the diagnostic criteria are clearly defined, the interpretation of histopathology slides may be subject to interobserver variability. The aim of this study was to determine the variability among dermatopathologists in the interpretation of clinically difficult melanocytic lesions. This study used the database of MelaFind®, a computer-vision system for the diagnosis of melanoma. All lesions were surgically removed and sent for independent evaluation by four dermatopathologists. Agreement was calculated using kappa statistics. A total of 1,249 pigmented melanocytic lesions were included. There was a substantial agreement among expert dermatopathologists: two-category kappa was 0.80 (melanoma vs. non-melanoma) and three-category kappa was 0.62 (malignant vs. borderline vs. benign melanocytic lesions). The agreement was significantly greater for patients ≥40 years (three-category kappa = 0.67) than for younger patients (kappa = 0.49). In addition, the agreement was significantly lower for patients with atypical mole syndrome (AMS) (kappa = 0.31) than for patients without AMS (kappa = 0.76). The data were limited by the inclusion/exclusion criteria of the MelaFind® study. This might represent a selection bias. The agreement was evaluated using kappa statistics. This is a standard method for evaluating agreement among pathologists, but might be considered controversial by some statisticians. Expert dermatopathologists have a high level of agreement when diagnosing clinically difficult melanocytic lesions. However, even among expert dermatopathologists, the current 'gold standard' is not perfect. Our results indicate that lesions from younger patients and patients with AMS may be more problematic for the dermatopathologists, suggesting that improved diagnostic criteria are needed for such patients.


Macronutrient intake and risk of urothelial cell carcinoma in the European prospective investigation into cancer and nutrition.
Previous studies have suggested that dietary factors may be important in the development of bladder cancer. We examined macronutrient intake in relation to risk of urothelial cell carcinoma among 469,339 men and women in the European Prospective Investigation into Cancer and Nutrition. Associations were examined using Cox regression, stratified by sex, age at recruitment and centre and further adjusted for smoking status and duration, body mass index and total energy intake. After an average of 11.3 years of follow-up, 1,416 new cases of urothelial cell carcinoma were identified. After allowing for measurement error, a 3% increase in the consumption of energy intake from animal protein was associated with a 15% higher risk (95% confidence interval [CI]: 3-30%; p(trend) = 0.01) and a 2% increase in energy from plant protein intake was associated with a 23% lower risk (95% CI: 36-7%, p(trend) = 0.006). Dietary intake of fat, carbohydrate, fibre or calcium was not associated with risk. These findings suggest that animal and/or plant protein may affect the risk of urothelial cell carcinoma, and examination of these associations in other studies is needed.


Enzymatic function of multiple origins regulates the progression of colorectal cancer and the development of metastases.
Enzymes play a crucial role in the progression of colorectal cancer and the development of metastases. They facilitate malignant cell invasion through the degradation of the extracellular matrix, the rupture of the basement membrane and the derangement of cell-cell adhesion. Furthermore, they promote tumour cell migration and support the evolution of metastatic lesions in the liver and other organs, through multiple molecular mechanisms, including growth factor release and angiogenesis. Urokinase plasminogen activator system, matrix metalloproteinases, heparanase and autocrine motility factor constitute important enzymatic complexes which assist colorectal cancer growth, with potential clinical applications in the diagnosis and treatment of the disease.


Effect of pistachio oil on gene expression of IFN-induced protein with tetratricopeptide repeats 2: a biomarker of inflammatory response.
When incorporated into the diet, pistachios have a beneficial effect on lipid and lipoprotein profiles. However, little is known about potential anti-inflammatory properties. This study was conducted to determine whether pistachio oil and an organic extract from pistachio oil extract (PE) regulated expression of inflammation-related genes. A mouse macrophage cell line (RAW 264.7 cells) was treated with pistachio oil and gene expression microarray analyses were performed. Pistachio oil significantly affected genes involved in immune response, defense response to bacteria, and gene silencing, of which INF-induced protein with tetratricopeptide repeats 2 (Ifit-2) was the most dramatically reduced. PE reduced the LPS-induced Ifit-2 by 78% and the bioactive molecules contained in PE, linoleic acid, and beta-sitosterol recapitulated this inhibition. Promoter analysis identified two adjacent IFN-stimulated response elements, which lie between -110 and -85bp of the 5'-flanking region of the Ifit-2 promoter, as being responsive to LPS activation and inhibition by PE. Our results indicate that pistachio oil and bioactive molecules present therein decrease Ifit-2 expressions, and due to the sensitivity of this effect, this gene is a potential biomarker for monitoring diet-induced changes in inflammation.


A PRDX1-p38α heterodimer amplifies MET-driven invasion of IDH-wildtype and IDH-mutant gliomas.
The Peroxiredoxin 1 (PRDX1) gene maps to chromosome arm 1p and is hemizygously deleted and epigenetically silenced in isocitrate dehydrogenase 1 or 2 (IDH)-mutant and 1p/19q-codeleted oligodendroglial tumors. In contrast, IDH-wildtype astrocytic gliomas including glioblastomas mostly lack epigenetic silencing and express PRDX1 protein. In our study, we investigated how PRDX1 contributes to the infiltrative growth of IDH-wildtype gliomas. Focusing on p38α-dependent pathways, we analyzed clinical data from 133 patients of the NOA-04 trial cohort to look for differences in the gene expression profiles of gliomas with wildtype or mutant IDH. Biochemical interaction studies as well as in vitro and ex vivo migration studies were used to establish a biological role of PRDX1 in maintaining pathway activity. Whole-brain high-resolution ultramicroscopy and survival analyses of pre-clinical mouse models for IDH-wildtype gliomas were then used for in vivo confirmation. Based on clinical data, we found that the absence of PRDX1 is associated with changes in the expression of MET/HGF signaling components. PRDX1 forms a heterodimer with p38α mitogen-activated protein kinase 14 (MAPK14), stabilizing phospho-p38α in glioma cells. This process amplifies hepatocyte growth factor (HGF)-mediated signaling and stimulates actin cytoskeleton dynamics that promote glioma cell migration. Whole-brain high-resolution ultramicroscopy confirms these findings, indicating that PRDX1 promotes glioma brain invasion in vivo. Finally, reduced expression of PRDX1 increased survival in mouse glioma models. Thus, our preclinical findings suggest that PRDX1 expression levels may serve as a molecular marker for patients who could benefit from targeted inhibition of MET/HGF signaling.


Cost-effectiveness of artemisinin-naphthoquine versus artemether-lumefantrine for the treatment of uncomplicated malaria in Papua New Guinean children.
A recent randomized trial showed that artemisinin-naphthoquine (AN) was non-inferior to artemether-lumefantrine (AL) for falciparum malaria and superior for vivax malaria in young Papua New Guinean children. The aim of this study was to compare the cost-effectiveness of these two regimens. An incremental cost-effectiveness analysis was performed using data from 231 children with Plasmodium falciparum and/or Plasmodium vivax infections in an open-label, randomized, parallel-group trial. Recruited children were randomized 1:1 to receive once daily AN for 3 days with water or twice daily AL for 3 days given with fat. World Health Organisation (WHO) definitions were used to determine clinical/parasitological outcomes. The cost of transport between the home and clinic, plus direct health-care costs, served as a basis for determining each regimen's incremental cost per incremental treatment success relative to AL by Day 42 and its cost per life year saved. In the usual care setting, AN was more effective for the treatment of uncomplicated malaria in children aged 0.5-5.9 years. AL and AN were equally efficacious for the treatment of falciparum malaria, however AN had increased anti-malarial treatment costs per patient of $10.46, compared with AL. AN was the most effective regimen for treatment of vivax malaria, but had increased treatment costs of $14.83 per treatment success compared with AL. Whilst AN has superior overall efficacy for the treatment of uncomplicated malaria in PNG children, AL was the less costly regimen. An indicative extrapolation estimated the cost per life year saved by using AN instead of AL to treat uncomplicated malaria to be $12,165 for girls and $12,469 for boys (discounted), which means AN may not be cost-effective and affordable for PNG at current cost. However, AN may become acceptable should it become WHO prequalified and/or should donated/subsidized drug supply become available.


Universality in stochastic exponential growth.
Recent imaging data for single bacterial cells reveal that their mean sizes grow exponentially in time and that their size distributions collapse to a single curve when rescaled by their means. An analogous result holds for the division-time distributions. A model is needed to delineate the minimal requirements for these scaling behaviors. We formulate a microscopic theory of stochastic exponential growth as a Master Equation that accounts for these observations, in contrast to existing quantitative models of stochastic exponential growth (e.g., the Black-Scholes equation or geometric Brownian motion). Our model, the stochastic Hinshelwood cycle (SHC), is an autocatalytic reaction cycle in which each molecular species catalyzes the production of the next. By finding exact analytical solutions to the SHC and the corresponding first passage time problem, we uncover universal signatures of fluctuations in exponential growth and division. The model makes minimal assumptions, and we describe how more complex reaction networks can reduce to such a cycle. We thus expect similar scalings to be discovered in stochastic processes resulting in exponential growth that appear in diverse contexts such as cosmology, finance, technology, and population growth.


The influence of the time interval between monoHER and doxorubicin administration on the protection against doxorubicin-induced cardiotoxicity in mice.
Despite its well-known cardiotoxicity, the anthracyclin doxorubicin (DOX) continues to be an effective and widely used chemotherapeutic agent. DOX-induced cardiac damage presumably results from the formation of free radicals by DOX. Reactive oxygen species particularly affect the cardiac myocytes because these cells seem to have a relatively poor antioxidant defense system. The semisynthetic flavonoid monohydroxyethylrutoside (monoHER) showed cardioprotection against DOX-induced cardiotoxicity through its radical scavenging and iron chelating properties. Because of the relatively short final half-life of monoHER (about 30 min), it is expected that the time interval between monoHER and DOX might be of influence on the cardioprotective effect of monoHER. Therefore, the aim of the present study was to investigate this possible effect. Six groups of 6 BALB/c mice were treated with saline, DOX alone or DOX (4 mg/kg i.v.) preceded by monoHER (500 mg/kg i.p.) with an interval of 10, 30, 60 or 120 min. After a 6-week treatment period and additional observation for 2 weeks, the mice were sacrificed. Their cardiac tissues were processed for light microscopy, after which cardiomyocyte damage was evaluated according to Billingham (in Cancer Treat Rep 62(6):865-872, 1978). Microscopic evaluation revealed that treatment with DOX alone induced significant cardiac damage in comparison to the saline control group (P<0.001). The number of damaged cardiomyocytes was 9.6-fold (95% CI 4.4-21.0) higher in mice treated with DOX alone than that in animals of the control group. The ratio of aberrant cardiomyocytes in mice treated with DOX preceded by monoHER and those in mice treated with saline ranged from 1.6 to 2.8 (mean 2.2, 95% CI 1.2-4.1, P=0.019). The mean protective effect by adding monoHER before DOX led to a significant 4.4-fold reduction (P<0.001, 95% CI 2.3-8.2) of abnormal cardiomyocytes. This protective effect did not depend on the time interval between monoHER and DOX administration (P=0.345). The results indicate that in an outpatient clinical setting monoHER may be administered shortly before DOX.


Crypt stem cell survival in the mouse intestinal epithelium is regulated by prostaglandins synthesized through cyclooxygenase-1.
Prostaglandins (PGs) are important mediators of epithelial integrity and function in the gastrointestinal tract. Relatively little is known, however, about the mechanism by which PGs affect stem cells in the intestine during normal epithelial turnover, or during wound repair. PGs are synthesized from arachidonate by either of two cyclooxygenases, cyclooxygenase-1 (Cox-1) or cyclooxygenase-2 (Cox-2), which are present in a wide variety of mamalian cells. Cox-1 is thought to be a constitutively expressed enzyme, and the expression of Cox-2 is inducible by cytokines or other stimuli in a variety of cell types. We investigated the role of PGs in mouse intestinal stem cell survival and proliferation following radiation injury. The number of surviving crypt stem cells was determined 3.5 d after irradiation by the microcolony assay. Radiation injury induced a dose-dependent decrease in the number of surviving crypts. Indomethacin, an inhibitor of Cox-1 and Cox-2, further reduced the number of surviving crypts in irradiated mice. The indomethacin dose response for inhibition of PGE2 production and reduction of crypt survival were similar. DimethylPGE2 reversed the indomethacin-induced decrease in crypt survival. Selective Cox-2 inhibitors had no effect on crypt survival. PGE2, Cox-1 mRNA, and Cox-1 protein levels all increase in the 3 d after irradiation. Immunohistochemistry for Cox-1 demonstrated localization in epithelial cells of the crypt in the unirradiated mouse, and in the regenerating crypt epithelium in the irradiated mouse. We conclude that radiation injury results in increased Cox-1 levels in crypt stem cells and their progeny, and that PGE2 produced through Cox-1 promotes crypt stem cell survival and proliferation.


Rare thyroid malignancies in Europe: Data from the information network on rare cancers in Europe (RARECAREnet).
Limited information is available on the incidence of rare thyroid cancer (TC) subtypes: anaplastic (ATC) and medullary (MTC). The aim of this study was to describe incidence variations and trends across European countries of all TC subtypes. We used the RARECAREnet database including 80721 TC incident cases in the period 2000-2007 from 77 population-based cancer registries (CRs) in Europe. In the trend analyses, we included 68890 TC cases from 53 CRs with at least 6 years of incidence data in the years 2000-2007. In Europe age-standardised incidence rates (ASR) in women were <0.3/100,000 for MTC and ATC whereas ASR were 5.3/100,000 for papillary thyroid cancer (PTC) and 1.1/100,000 for follicular TC (FTC). Corresponding ASRs in men were <0.2/100,000 for MTC and ATC, 1.5 for PTC and 0.4 for FTC. Across countries and in both sexes the incidence of FTC and MTC was moderately correlated (r ~ 0.5) with that of PTC, while a less marked correlation (r < 0.4) emerged for ATC ASRs. The changes of the PTC ASRs across countries and time were weakly (r < 0.3) or moderately (r ~ 0.5) correlated to the changes of the other subtypes for both sexes. The huge increase and heterogeneity between countries of PTC incidence has a small influence on the trends and variations of MTC and ATC in Europe. Large-scale epidemiological and clinical registry-based studies are warranted to increase knowledge about the rarest TC subtypes. This information would be fundamental for the design of new clinical trials and for inference.


Catalyzing the scale-up of community-based primary healthcare in a rural impoverished region of northern Ghana.
Ghana's Community-based Health Planning and Services (CHPS) initiative develops accessible healthcare with participatory community support, using strategies developed and tested by a project of the Navrongo Health Research Centre. In 1996, the project was expanded to a district-wide four-celled trial. In response to evidence that strategies could reduce fertility and childhood mortality, a replication project was launched to develop methods for scale-up. Based on experience gained, CHPS scale-up was launched in 2000. Although CHPS now reaches all of Ghana's districts, the pace of scale-up within districts has been slow. In response, the Ministry of Health conducted a review of factors that constrain CHPS scale-up and problems that detract from its original evidence-based design. To resolve problems that were identified, a project was launched in 2010 to test means of accelerating CHPS scale-up and expand its range of care. Known as the Ghana Essential Health Interventions Program (GEHIP), the project provided catalytic revenue to four treatment district managers for 3 years, in conjunction with implementation of strategies for comprehensive leadership development and community partnership. Monitoring systems were developed to gauge CHPS coverage time trends in all nine study districts. GEHIP successfully accelerated CHPS implementation, producing 100% of its targeted community coverage within 5 years of implementation. Coverage in comparison districts also improved. However, the rate of coverage and per cent of the population reached by CHPS in comparison districts was only half that of GEHIP districts. GEHIP success in completing CHPS coverage represents the initial stage of a national program for strengthening community health systems in Ghana. Copyright © 2015 John Wiley & Sons, Ltd.


New Generation of Orally Disintegrating Tablets for Sustained Drug Release: A Propitious Outlook.
Orally disintegrating tablets (ODTs) or orodispersible tablets are solid dosage forms that disintegrate within 3 minutes in the mouth into a paste that can be easily swallowed. ODTs have improved over the past years, in an attempt to produce a safe and efficient substitute to the conventional oral dosage forms, particularly for dysphagia patients. Since its introduction in the market in the 1980s, ODTs expanded rapidly and achieved revenues over $3 billion in 2006 and sustaining 20% annual growth. It is therefore evident that ODTs carry good commercial value, however there is potential for improvement. Current sustained-release technologies may be exploited and incorporated into an ODT to provide greater therapeutic value by reducing the need for multiple daily dosing regimens and improving patient adherence. A number of technologies such as polymer coated nanoparticles, stimuli-responsive polymers and ion-exchange resins have emerged to produce robust, sustained release orally disintegrating tablets (SR-ODT). The purpose of this review is to highlight these various approaches and techniques and how they have been utilised in an ODT formulation to extend differentiated line, market exclusivity and patent life. The review also explores future perspective and the potential challenges that SR-ODTs will face.


Clinical significance of the isolation of Staphylococcus epidermidis from bone biopsy in diabetic foot osteomyelitis.
Coagulase-negative staphylococci are considered as microorganisms with little virulence and usually as contaminants. In order to establish the role of Staphylococcus epidermidis as a pathogen in diabetic foot osteomyelitis, in addition to the isolation of the sole bacterium from the bone it will be necessary to demonstrate the histopathological changes caused by the infection. A consecutive series of 222 diabetic patients with foot osteomyelitis treated surgically in the Diabetic Foot Unit at La Paloma Hospital (Las Palmas de Gran Canaria, Canary Islands, Spain) between 1 October 2002 and 31 October 2008. From the entire series including 213 bone cultures with 241 isolated organisms, we have analyzed only the 139 cases where Staphylococci were found. We analyzed several variables between the two groups: Staphylococcus aureus versus Staphylococcus epidermidis. Of the 134 patients included in this study, Staphlylococcus epidermidis was found as the sole bacterium isolated in 11 cases and accompanied by other bacteria in 12 cases. Staphlylococcus aureus was found as the sole bacterium isolated in 72 cases and accompanied by other bacteria in 39 cases. Histopathological changes were found in the cases of osteomyelitis where Staphylococcus epidermidis was the sole bacterium isolated. Acute osteomyelitis was found to a lesser extent when Staphylococcus epidermidis was the sole bacterium isolated but without significant differences with the cases where Staphylococcus aureus was the sole bacterium isolated. Staphylococcus epidermidis should be considered as a real pathogen, not only a contaminant, in diabetic patients with foot osteomyelitis when the bacterium is isolated from the bone. No differences in the outcomes of surgical treatment have been found with cases which Staphlylococcus aureus was isolated.


Sex-specific brain erythropoietin regulation of mouse metabolism and hypothalamic inflammation.
The blood hormone erythropoietin (EPO), upon binding to its receptor (EpoR), modulates high-fat diet-induced (HFD-induced) obesity in mice, improves glucose tolerance, and prevents white adipose tissue inflammation. Transgenic mice with constitutive overexpression of human EPO solely in the brain (Tg21) were used to assess the neuroendocrine EPO effect without increasing the hematocrit. Male Tg21 mice resisted HFD-induced weight gain; showed lower serum adrenocorticotropic hormone, corticosterone, and C-reactive protein levels; and prevented myeloid cell recruitment to the hypothalamus compared with WT male mice. HFD-induced hypothalamic inflammation (HI) and microglial activation were higher in male mice, and Tg21 male mice exhibited a lower increase in HI than WT male mice. Physiological EPO function in the brain also showed sexual dimorphism in regulating HFD response. Female estrogen production blocked reduced weight gain and HI. Targeted deletion of EpoR gene expression in neuronal cells worsened HFD-induced glucose intolerance in both male and female mice but increased weight gain and HI in the hypothalamus in male mice only. Both male and female Tg21 mice kept on normal chow and HFD showed significantly improved glycemic control. Our data indicate that cerebral EPO regulates weight gain and HI in a sex-dependent response, distinct from EPO regulation of glycemic control, and independent of erythropoietic EPO response.


Attractants from Bartlett pear for codling moth, Cydia pomonella (L.), larvae.
The alkyl ethyl and methyl esters of (2E,4Z)-2,4-decadienoic acid found in head-space samples of ripe Bartlett pear (Pyrus communis L.) stimulated a response from neonate larvae of the codling moth (CM), Cydia pomonella (L.), in both static-air Petri-plate and in upwind Y-tube and straight-tube olfactometer bioassays. In comparison with the known CM neonate attractant, (E,E)-alpha-farnesene, ethyl (2E,4Z)-2,4-decadienoate was attractive at 10-fold and 1,000-fold lower threshold dosages in the Petri-plate and in the Y-tube bioassays, respectively. Methyl (2E,4Z)-2,4-decadienoate was attractive to CM neonates in these bioassays at much higher doses than ethyl (2E,4Z)-2,4-decadienoate. Other principal head-space volatiles from ripe pear fruit and pear leaves, including butyl acetate, hexyl acetate, (Z)-3-hexenyl acetate, and (E)-beta-ocimene, were not attractive to CM neonates. The potential uses of these pear kairomones for monitoring and control of CM in walnuts and apple are discussed.


Clinicians' Ability to Detect a Palpable Difference in Spinal Stiffness Compared With a Mechanical Device.
The purpose of this study was to quantify the threshold at which clinicians can detect a difference in spinal stiffness of the thoracic and lumbar spine via palpation and then determine if this detection threshold would affect a clinician's ability to identify changes in spinal stiffness as measured by an objective instrument. In this study, the threshold at which a change in spinal stiffness was detected was quantified in 12 experienced clinicians (physical therapists and doctors of chiropractic) by changing the differential stiffness in 2 inflatable targets until the clinician could no longer identify which was stiffer. In the second part of the study, clinicians then were asked to palpate pre-identified pairs of vertebrae in an asymptomatic volunteer and to identify the stiffer of the pair (T7 and L3, T7 and L4, L3 and L4), and the biomechanical stiffness of each vertebral pair was quantified objectively by a validated instrument. The mean stiffness detection threshold for the clinicians was 8%. Objective measurement of the stiffness differential between vertebral pairs was 30% for T7* and L3, 20% for T7* and L4, and 10% for L3* and L4 (*denotes the stiffer of the pair). Ten of 12 clinicians correctly identified T7 as stiffer when compared with L3 and T7 as stiffer than L4. Alternatively, when the differential vertebral pair stiffness was similar to the stiffness detection threshold (~8%), clinicians were less successful in identifying the stiffer vertebra of the pair; 4 of 12 clinicians correctly identified L3 as being stiffer compared with L4. These results suggest that the physiological limits of human palpation may limit the ability of clinicians to identify small alterations in spine stiffness.


Hydrophobic distal pocket affects NO-heme geminate recombination dynamics in dehaloperoxidase and H64V myoglobin.
The recombination dynamics of NO with dehaloperoxidase (DHP) from Amphitrite ornata following photolysis were measured by femtosecond time-resolved absorption spectroscopy. Singular value decomposition (SVD) analysis reveals two important basis spectra. The first SVD basis spectrum reports on the population of photolyzed NO molecules and has the appearance of the equilibrium difference spectrum between the deoxy and NO forms of DHP. The first basis time course has two kinetic components with time constants of tau(11) approximately 9 ps and tau(12) approximately 50 ps that correspond to geminate recombination. The fast geminate process tau(11) arises from a contact pair with the heme iron in a bound state with S = 3/2 spin. The slow geminate process tau(12) corresponds to the recombination from a more remote docking site >3 A from the heme iron with the greater barrier corresponding to a S = 5/2 spin state. The second SVD basis spectrum represents a time-dependent Soret band shift indicative of heme photophysical processes and protein relaxation with time constants of tau(21) approximately 3 ps and tau(22) approximately 17 ps, respectively. A comparison between the more rapid rate constant of the slow geminate phase in DHP-NO and horse heart myoglobin (HHMbNO) or sperm whale myoglobin (SWMbNO) suggests that protein interactions with photolyzed NO are weaker in DHP than in the wild-type MbNOs, consistent with the hydrophobic distal pocket of DHP. The slower protein relaxation rate tau(22) in DHP-NO relative to HHMbNO implies less effective trapping in the docking site of the distal pocket and is consistent with a greater yield for the fast geminate process. The trends observed for DHP-NO also hold for the H64V mutant of SWMb (H64V MbNO), consistent with a more hydrophobic distal pocket for that protein as well. We examine the influence of solution viscosity on NO recombination by varying the glycerol content in the range from 0% to 90% (v/v). The dominant effect of increasing viscosity is the increase of the rate of the slow geminate process, tau(12), coupled with a population decrease of the slow geminate component. Both phenomena are similar to the effect of viscosity on wild-type Mb due to slowing of protein relaxation resulting from an increased solution viscosity and protein surface dehydration.


Does neighborhood violence lead to depression among caregivers of children with asthma?
Prior studies have related community violence to depression among children, but few studies have examined this relationship among adults. We hypothesized that victimization, awareness, and fear of neighborhood violence would increase the odds of depression among adult caregivers of children with asthma. We surveyed caregivers in the Baltimore Indoor Environment Study of Asthma in Kids (BIESAK), USA. The primary outcome was screening positive for depression on the Center for Epidemiological Studies Depression index. We assessed victimization, awareness, and fear of neighborhood violence, and conducted spatial analysis identifying subject homes within 500 ft of a homicide to validate survey measures of neighborhood violence. A multilevel logistic model with clustering by neighborhood estimated odds ratios and 95% confidence intervals. Survey responses about fear of neighborhood violence were strongly predicted by having a home within 500 ft of a homicide. Of 150 caregivers of children with asthma, 49% were aware of a neighborhood violent event, 36% were fearful of neighborhood violence, 22% reported victimization, and 27% had a homicide within 500 ft of the home. In our multilevel model, fear of violence increased the odds of depression by 6.7. Victimization was associated with a possible trend towards depression, and awareness of neighborhood violence did not increase the odds of depression. Based on our findings, personal experience with neighborhood violence may be more important than simple awareness. Health care workers should consider screening for depression among patients exposed to community violence.


Molecular cloning of the canine telomerase reverse transcriptase gene and its expression in neoplastic and non-neoplastic cells.
To perform molecular cloning of the canine telomerase reverse transcriptase (TERT) gene and determine its expression in neoplastic and non-neoplastic cells. 9 canine tumor cell lines derived from various neoplasms, 16 primary canine tumors, and tissues from 15 normal canine organs. Tumor cell lines were derived from canine tumors that included osteosarcoma, mammary gland adenocarcinoma, melanoma, acute lymphoblastic leukemia, lymphoma, and mastocytoma and a canine primary fibroblast culture. Canine TERT complementary DNA (cDNA) was amplified by use of polymerase chain reaction (PCR) and sequenced. Expression of TERT mRNA was examined by reverse transcription (RT)-PCR assay. Telomerase activity was measured by use of the telomeric repeat amplification protocol assay. The canine TERT cDNA clone was 237 base pairs in length and contained a central region encoding the reverse transcriptase motif 2. Expression of TERT mRNA was detected in canine tumor cell lines that had telomerase activity but not in telomerase-negative canine primary fibroblasts. The TERT mRNA was detected in 13 of 16 canine tumor tissues and several normal tissues such as liver, ovary, lymph node, and thymus. A significant correlation between TERT expression level and telomerase activity was noted. Expression of TERT mRNA was closely associated with telomerase activity in neoplastic cells as well as some non-neoplastic cells from dogs. In addition to telomerase activity, expression of TERT mRNA can be used as a marker of tumor cells.


Loss of miR-29a/b1 promotes inflammation and fibrosis in acute pancreatitis.
MicroRNA-29 (miR-29) is a critical regulator of fibroinflammatory processes in human diseases. In this study, we found a decrease in miR-29a in experimental and human chronic pancreatitis, leading us to investigate the regulatory role of the miR-29a/b1 cluster in acute pancreatitis (AP) utilizing a conditional miR-29a/b1-KO mouse model. miR-29a/b1-sufficient (WT) and -deficient (KO) mice were administered supramaximal caerulein to induce AP and characterized at different time points, utilizing an array of IHC and biochemical analyses for AP parameters. In caerulein-induced WT mice, miR-29a remained dramatically downregulated at injury. Despite high-inflammatory milieu, fibrosis, and parenchymal disarray in the WT mice during early AP, the pancreata fully restored during recovery. miR-29a/b1-KO mice showed significantly greater inflammation, lymphocyte infiltration, macrophage polarization, and ECM deposition, continuing until late recovery with persistent parenchymal disorganization. The increased pancreatic fibrosis was accompanied by enhanced TGFβ1 coupled with persistent αSMA+ PSC activation. Additionally, these mice exhibited higher circulating IL-6 and inflammation in lung parenchyma. Together, this collection of studies indicates that depletion of miR-29a/b1 cluster impacts the fibroinflammatory mechanisms of AP, resulting in (a) aggravated pathogenesis and (b) delayed recovery from the disease, suggesting a protective role of the molecule against AP.


Tubulin subunits exist in an activated conformational state generated and maintained by protein cofactors.
The production of native alpha/beta tubulin heterodimer in vitro depends on the action of cytosolic chaperonin and several protein cofactors. We previously showed that four such cofactors (termed A, C, D, and E) together with native tubulin act on beta-tubulin folding intermediates generated by the chaperonin to produce polymerizable tubulin heterodimers. However, this set of cofactors generates native heterodimers only very inefficiently from alpha-tubulin folding intermediates produced by the same chaperonin. Here we describe the isolation, characterization, and genetic analysis of a novel tubulin folding cofactor (cofactor B) that greatly enhances the efficiency of alpha-tubulin folding in vitro. This enabled an integrated study of alpha- and beta-tubulin folding: we find that the pathways leading to the formation of native alpha- and beta-tubulin converge in that the folding of the alpha subunit requires the participation of cofactor complexes containing the beta subunit and vice versa. We also show that sequestration of native alpha-or beta-tubulins by complex formation with cofactors results in the destabilization and decay of the remaining free subunit. These data demonstrate that tubulin folding cofactors function by placing and/or maintaining alpha-and beta-tubulin polypeptides in an activated conformational state required for the formation of native alpha/beta heterodimers, and imply that each subunit provides information necessary for the proper folding of the other.


Electron-Beam Induced Transformations of Layered Tin Dichalcogenides.
By combining high-resolution transmission electron microscopy and associated analytical methods with first-principles calculations, we study the behavior of layered tin dichalcogenides under electron beam irradiation. We demonstrate that the controllable removal of chalcogen atoms due to electron irradiation, at both room and elevated temperatures, gives rise to transformations in the atomic structure of Sn-S and Sn-Se systems so that new phases with different properties can be induced. In particular, rhombohedral layered SnS2 and SnSe2 can be transformed via electron beam induced loss of chalcogen atoms into highly anisotropic orthorhombic layered SnS and SnSe. A striking dependence of the layer orientation of the resulting SnS-parallel to the layers of ultrathin SnS2 starting material, but slanted for transformations of thicker few-layer SnS2-is rationalized by a transformation pathway in which vacancies group into ordered S-vacancy lines, which convert via a Sn2S3 intermediate to SnS. Absence of a stable Sn2Se3 intermediate precludes this pathway for the selenides, hence SnSe2 always transforms into basal plane oriented SnSe. Our results provide microscopic insights into the transformation mechanism and show how irradiation can be used to tune the properties of layered tin chalcogenides for applications in electronics, catalysis, or energy storage.


[Long-term results of calcaneal fracture treatment by open reduction and internal fixation using a calcaneal locking compression plate from an extended lateral approach].
To report on the surgical treatment of intra-articular calcaneal fractures by open reduction and internal fixation with a calcaneal locking compression plate (LCP) from an extended lateral approach, and to retrospectively analyze the mid-term results in a group of patients treated by this technique. In the period from August 2005 till March 2007, a total of 49 patients with 61 calcaneal fractures were treated. Of these, 11 (18 %) were treated conservatively. Reduction combined with Kirschner-wire fixation was used in four fractures (6.6 %). Open reduction with internal calcaneal LCP fixation (ORIF- calcaneal LCP) from an extended lateral approach was carried out to treat 46 fractures (75.4 %) in 38 patients. The group evaluated here comprised 29 patients with 33 calcaneal fractures treated by ORIF-calcaneal LCP at a follow-up longer than 6 months. The fractures were classified on the basis of computer tomography (CT) findings as Sanders types I to IV. The group had two woman (6.9 %) and 27 men (93.1 %) with an average age of 34.2 years (range, 19-55 years). In 11 fractures (33.3 %), the primary treatment included filling a central cancellous bone defect area. Calcium phosphate bone substitute material (resorbable ChronOS) was used in nine cases (27.3 %), a self-solidifying hydroxyapatite implant was injected in two (6.1 %) cases (X3 Wright and Norian SRS, respectively), and a bone allograft was implanted in one case (3 %). Indicated for surgery were patients with an intra-articular calcaneal fracture, Sanders type II or type III, with articular surface displacement by more than 1 mm. Contraindications included age over sixty years, poor cooperation, smoking habits, peripheral vascular disease or skin infection. Surgery was performed only after oedema had resolved. The aim of our treatment was to achieve anatomical reconstruction of all articular surfaces, to restore the height, length, width and axis of the heel bone, to carry out primary stable osteosynthesis, and to enable the patient to begin rehabilitation with early mobilization. A passive rehabilitation usually started on the first post-operative day, and full weight-bearing of the extremity was allowed not earlier than 3 months post-operatively. Regular check-ups were at 6 weeks, 3, 6 and 12 months and then every year. The mid-term results were evaluated by the system of Rowe et al., scoring rest pain, possibility of return to preinjury jobs, use of walking aids, restriction of physical activity and limping. RESULTS The most frequent cause of injury was a fall or jump from height; this was recorded in 27 patients (93.1 %). An open fracture was diagnosed on two occasions (6.9 %). Bilateral calcaneal fractures were found in six patients (20.7 %); four (13.8 %) were treated by bilateral ORIF-calcaneal LPC and two (6.9 %) underwent closed reduction on one and Kirschnerwire transfixation on the other extremity. A combined injury to the musculoskeletal system was diagnosed in 11 patients (38 %), in whom four (13.8 %) had a tibial pylon fracture of the contralateral limb and four (13.8 %) had a thoracolumbar spine fracture. The surgical procedure was performed on average within 11.7 days of injury, and the average hospital stay was 18.2 days (range, 6 to 18 days). Early post-operative complications were recorded in six patients (20.7 %). Wound dehiscence was found in two (6.9 %), necrosis of wound edges in two (6.9 %), and early superficial infection responding to antibiotic therapy also in two patients (6.9 %). Deep infection, non-union or post-operative compartment syndrome were not recorded. Excellent Rowe scores were achieved in 10 patients (34.5 %), good in 15 (51.7 %) and satisfactory in two (6.9 %). Only two patients (6.9 %) reported poor outcome. The methods of classification and treatment of calcaneal fractures continue to be a frequently discussed topic. The technique of ORIF-calcaneal LCP from an extended lateral approach has recently been preferred for patients with displaced Sanders type II or III calcaneal fractures. In agreement with other literature references, this approach allowed us to observe the fracture, to reduce both the subtalar and calcaneocuboid articulations, to stabilize the fracture by internal fixation and to begin early rehabilitation. Because of the risk of early complications, the timing of surgery and a thorough consideration of indications and contraindications are of principal importance.We agree with other authors that filling calcaneal bone defects is not necessary. Pre- and post-operative CT scans are necessary. In accordance with literature data, fractures developing compartment syndrome are indicated for urgent fasciotomy and ORIF-calcaneal LCP should be postponed. In patients with multiple trauma and also in those with open calcaneal fractures, a temporary stabilization with an external fixator medially is performed first, and then converted to a second-stage, ORIF-LCP procedure. The surgical treatment of displaced intra-articular fractures that involves open reduction from an extended lateral approach and internal fixation with a calcaneal LCP shows very good preliminary results. A CT examination is required for the diagnosis and classification of fractures and a correct indication for surgery. Good timing is of principal importance. An urgent surgical intervention is necessary in open fractures or in those in which soft tissues are squashed by bone fragments. In the other fractures, surgery is carried out after oedema subsidence. Foot compartment syndrome is a serious complication of calcaneal fractures and urgent fasciotomy is the only adequate therapy. Full weight-bearing of the operated extremity depends on the rate of bone healing; it is usually allowed at 3 months after surgery.


Probabilistic modeling of sewer deterioration using inspection data.
Accurate prediction of current and future conditions of sewer systems is crucial to manage the sewer system wisely, cost-effectively and efficiently. The application of historical databases of visual inspection data to sewer deterioration modeling seems common sense. However, in The Netherlands, sewer inspection data is only used to determine the direct need for rehabilitation. This paper outlines the possibilities of using inspection data for deterioration modeling and discusses the problems encountered. A case study was performed on the modeling of the condition aspect 'surface damage by corrosion or mechanical action' using a Markov model.


Commuting and Sleep: Results From the Hispanic Community Health Study/Study of Latinos Sueño Ancillary Study.
Commute time is associated with reduced sleep time, but previous studies have relied on self-reported sleep assessment. The present study investigated the relationships between commute time for employment and objective sleep patterns among non-shift working U.S. Hispanic/Latino adults. From 2010 to 2013, Hispanic/Latino employed, non-shift-working adults (n=760, aged 18-64 years) from the Sueño study, ancillary to the Hispanic Community Health Study/Study of Latinos, reported their total daily commute time to and from work, completed questionnaires on sleep and other health behaviors, and wore wrist actigraphs to record sleep duration, continuity, and variability for 1 week. Survey linear regression models of the actigraphic and self-reported sleep measures regressed on categorized commute time (short: 1-44 minutes; moderate: 45-89 minutes; long: ≥90 minutes) were built adjusting for relevant covariates. For associations that suggested a linear relationship, continuous commute time was modeled as the exposure. Moderation effects by age, sex, income, and depressive symptoms also were explored. Commute time was linearly related to sleep duration on work days such that each additional hour of commute time conferred 15 minutes of sleep loss (p=0.01). Compared with short commutes, individuals with moderate commutes had greater sleep duration variability (p=0.04) and lower interdaily stability (p=0.046, a measure of sleep/wake schedule regularity). No significant associations were detected for self-reported sleep measures. Commute time is significantly associated with actigraphy-measured sleep duration and regularity among Hispanic/Latino adults. Interventions to shorten commute times should be evaluated to help improve sleep habits in this minority population.


Quantitative differences in adhesiveness of type 1 fimbriated Escherichia coli due to structural differences in fimH genes.
Type 1 fimbriae are heteropolymeric surface organelles responsible for the D-mannose-sensitive (MS) adhesion of Escherichia coli. We recently reported that variation of receptor specificity of type 1 fimbriae can result solely from minor alterations in the structure of the gene for the FimH adhesin subunit. To further study the relationship between allelic variation of the fimH gene and adhesive properties of type 1 fimbriae, the fimH genes from five additional strains were cloned and used to complement the FimH deletion in E. coli KB18. When the parental and recombinant strains were tested for adhesion to immobilized mannan, a wide quantitative range in the ability of bacteria to adhere was noted. The differences in adhesion do not appear to be due to differences in the levels of fimbriation or relative levels of incorporation of FimH, because these parameters were similar in low-adhesion and high-adhesion strains. The nucleotide sequence for each of the fimH genes was determined. Analysis of deduced FimH sequences allowed identification of two sequence homology groups, based on the presence of Asn-70 and Ser-78 or Ser-70 and Asn-78 residues. The consensus sequences for each group conferred very low adhesion activity, and this low-adhesion phenotype predominated among a group of 43 fecal isolates. Strains isolated from a different host niche, the urinary tract, expressed type 1 fimbriae that conferred an increased level of adhesion. The results presented here strongly suggest that the quantitative variations in MS adhesion are due primarily to structural differences in the FimH adhesin. The observed differences in MS adhesion among populations of E. coli isolated from different host niches call attention to the possibility that phenotypic variants of FimH may play a functional role in populations dynamics.


On the parameter optimization of Support Vector Machines for binary classification.
Classifying biological data is a common task in the biomedical context. Predicting the class of new, unknown information allows researchers to gain insight and make decisions based on the available data. Also, using classification methods often implies choosing the best parameters to obtain optimal class separation, and the number of parameters might be large in biological datasets. Support Vector Machines provide a well-established and powerful classification method to analyse data and find the minimal-risk separation between different classes. Finding that separation strongly depends on the available feature set and the tuning of hyper-parameters. Techniques for feature selection and SVM parameters optimization are known to improve classification accuracy, and its literature is extensive. In this paper we review the strategies that are used to improve the classification performance of SVMs and perform our own experimentation to study the influence of features and hyper-parameters in the optimization process, using several known kernels.


Characterizaiton of two populations of human lymphocytes bearing easily detectable surface immunoglobulin.
Two separate lymphocyte populations, each bearing easily detectable surface immunoglobulin, have been detected in human peripheral blood. The first, B cells, has surface determinants that are stable at 37degreeC, but are removed by pronase and regenerate in culture. The cells are nylon adherent and have a receptor for C3, and studies with unit gravity sedimentation indicate they are mostly small lymphocytes. B cells comprise 9.5% of the total lymphocytes, with the normal range from 3-16%. As many or more lymphocytes lack membrane-incorporated Ig determinants but have an Fc receptor that binds IgG1 and IgG3 in normal serum maximally at 4degreeC. This receptor for cytophilic IgG is removed by pronase but not by trypsin. The second population has been named L lymphocytes because of membrane-labile IgG determinants. L cells do not adhere to nylon, do not form rosettes with sheep erythrocytes sensitized with antibody and mouse complement, and are larger than small lymphocytes. These lymphocytes with cold-reactive Fc receptors for serum IgG do not form E-rosettes or respond to phytohemaggutinin. Since L cells do not have surface markers of T and B lymphocytes, it is likely that they comprise a separate population.


The therapeutic use of the relaxation response in stress-related diseases.
The objective of this work was to investigate a possible (therapeutic) connection between the relaxation response (RR) and stress-related diseases. Further, common underlying molecular mechanisms and autoregulatory pathways were examined. For the question of (patho)physiology and significance of RR techniques in the treatment of stress-related diseases, we analyzed peer-reviewed references only. The RR has been shown to be an appropriate and relevant therapeutic tool to counteract several stress-related disease processes and certain health-restrictions, particularly in certain immunological, cardiovascular, and neurodegenerative diseases/mental disorders. Further, common underlying molecular mechanisms may exist that represent a connection between the stress response, pathophysiological findings in stress-related diseases, and physiological changes/autoregulatory pathways described in the RR. Here, constitutive or low-output nitric oxide (NO) production may be involved in a protective or ameliorating context, whereas inducible, high-output NO release may facilitate detrimental disease processes. In mild or early disease states, a high degree of biological and physiological flexibility may still be possible (dynamic balance). Here, the therapeutic use of RR techniques may be considered particularly relevant, and the observable (beneficial) effects may be exerted via activation of constitutive NO pathways. RR techniques, regularly part of professional stress management or mind/body medical settings, represent an important tool to be added to therapeutic strategies dealing with stress-related diseases. Moreover, as part of 'healthy' life-style modifications, they may serve primary (or secondary) prevention. Further studies are necessary to elucidate the complex physiology underlying the RR and its impact upon stress-related disease states.


Alterations in the gut microbiome implicate key taxa and metabolic pathways across inflammatory arthritis phenotypes.
Musculoskeletal diseases affect up to 20% of adults worldwide. The gut microbiome has been implicated in inflammatory conditions, but large-scale metagenomic evaluations have not yet traced the routes by which immunity in the gut affects inflammatory arthritis. To characterize the community structure and associated functional processes driving gut microbial involvement in arthritis, the Inflammatory Arthritis Microbiome Consortium investigated 440 stool shotgun metagenomes comprising 221 adults diagnosed with rheumatoid arthritis, ankylosing spondylitis, or psoriatic arthritis and 219 healthy controls and individuals with joint pain without an underlying inflammatory cause. Diagnosis explained about 2% of gut taxonomic variability, which is comparable in magnitude to inflammatory bowel disease. We identified several candidate microbes with differential carriage patterns in patients with elevated blood markers for inflammation. Our results confirm and extend previous findings of increased carriage of typically oral and inflammatory taxa and decreased abundance and prevalence of typical gut clades, indicating that distal inflammatory conditions, as well as local conditions, correspond to alterations to the gut microbial composition. We identified several differentially encoded pathways in the gut microbiome of patients with inflammatory arthritis, including changes in vitamin B salvage and biosynthesis and enrichment of iron sequestration. Although several of these changes characteristic of inflammation could have causal roles, we hypothesize that they are mainly positive feedback responses to changes in host physiology and immune homeostasis. By connecting taxonomic alternations to functional alterations, this work expands our understanding of the shifts in the gut ecosystem that occur in response to systemic inflammation during arthritis.


Synthesis and antinociceptive-antimicrobial activities of some new amide derivatives of 3,5-di/-and 1,3,5-trimethylpyrazoles.
Some N-(3,5-di-/1,3,5-trimethylpyrazole-4-yl)-4-substitutedbenzamide derivatives were prepared as possible antiociceptive-antimicrobial agents. New amide derivatives (3-12) were synthesized by reacting 4-amino-3,5-di and 1,3,5-trimethylpyrazoles with 4-substitutedbenzoyl chlorides. Hotplate and tail-immersion tests were used for the determination of the antinociceptive activity. Morphine, was used as a standard test drug. All compounds were administered at a dose of 100 mg/kg ip and some of them had significant antinociceptive activity in both tests. Compound 10 (N-(1,3,5-trimethylpyrazole-4-yl)-4-bromobenzamide), was the most active one in both tests among the compounds. The antinociceptive activity of the compounds 10, 11 (N-(1,3,5-trimethylpyrazole-4-yl)-4-chlorobenzamide), and 12 (N-(1,3,5-trimethylpyrazole-4-yl)-4-fluorobenzamide), started at 30 minutes and continued up to 150 minutes in the hotplate test. Also compounds were tested for their in vitro antimicrobial activity, but exhibited weak antibacterial activity.


Selectivity of C3-opsonin targeted complement inhibitors: A distinct advantage in the protection of erythrocytes from paroxysmal nocturnal hemoglobinuria patients.
Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by complement-mediated cell lysis due to deficiency of GPI-anchored complement regulators. Blockage of the lytic pathway by eculizumab is the only available therapy for PNH patients and shows remarkable benefits, but regularly yields PNH erythrocytes opsonized with fragments of complement protein C3, rendering such erythrocytes prone to extravascular hemolysis. This effect is associated with insufficient responsiveness seen in a subgroup of PNH patients. Novel C3-opsonin targeted complement inhibitors act earlier in the cascade, at the level of activated C3 and are engineered from parts of the natural complement regulator Factor H (FH) or complement receptor 2 (CR2). This inhibitor class comprises three variants of "miniFH" and the clinically developed "FH-CR2" fusion-protein (TT30). We show that the approach of FH-CR2 to target C3-opsonins was more efficient in preventing complement activation induced by foreign surfaces, whereas the miniFH variants were substantially more active in controlling complement on PNH erythrocytes. Subtle differences were noted in the ability of each version of miniFH to protect human PNH cells. Importantly, miniFH and FH-CR2 interfered only minimally with complement-mediated serum killing of bacteria when compared to untargeted inhibition of all complement pathways by eculizumab. Thus, the molecular design of each C3-opsonin targeted complement inhibitor determines its potency in respect to the nature of the activator/surface providing potential functionality in PNH.


Preparation of (Zn1+xGe)(N2Ox) nanoparticles with enhanced NOx decomposition activity under visible light irradiation by nitridation of Zn2GeO4 nanoparticles designed precisely.
Quaternary zinc germanium oxynitride (Zn1+xGe)(N2Ox), a solid solution between ZnGeN2 and ZnO with a wurtzite structure, is one of the attractive photocatalysts under visible-light irradiation. In this study, the synthesis of (Zn1+xGe)(N2Ox) nanoparticles was achieved by the nitridation of Zn2GeO4 nanoparticles designed precisely to enhance their photocatalytic NOx decomposition activity under both UV and visible light irradiation. The obtained (Zn1+xGe)(N2Ox) nanoparticles exhibited a high specific surface area and visible light absorption induced by the narrow band gap of ca. 2.6-2.8 eV, both of which are reasons for the enhancement of photocatalytic activity. The oxide precursors with a nanoparticle morphology were prepared by a facile solvothermal method with various volumes of TEA (triethanolamine) as an additive. The relationships of nitridation time and TEA volume in the solvothermal reaction for the synthesis of the precursor with morphology, specific surface area, and photocatalytic NOx decomposition activity of the nitrided samples were investigated. The increase of active sites by the high surface area and the enhanced visible-light absorption ability as well as the defect amounts and states can be largely related to the excellent NOx decomposition activity of (Zn1+xGe)(N2Ox).


Toxicity of the main electronic cigarette components, propylene glycol, glycerin, and nicotine, in Sprague-Dawley rats in a 90-day OECD inhalation study complemented by molecular endpoints.
While the toxicity of the main constituents of electronic cigarette (ECIG) liquids, nicotine, propylene glycol (PG), and vegetable glycerin (VG), has been assessed individually in separate studies, limited data on the inhalation toxicity of them is available when in mixtures. In this 90-day subchronic inhalation study, Sprague-Dawley rats were nose-only exposed to filtered air, nebulized vehicle (saline), or three concentrations of PG/VG mixtures, with and without nicotine. Standard toxicological endpoints were complemented by molecular analyses using transcriptomics, proteomics, and lipidomics. Compared with vehicle exposure, the PG/VG aerosols showed only very limited biological effects with no signs of toxicity. Addition of nicotine to the PG/VG aerosols resulted in effects in line with nicotine effects observed in previous studies, including up-regulation of xenobiotic enzymes (Cyp1a1/Fmo3) in the lung and metabolic effects, such as reduced serum lipid concentrations and expression changes of hepatic metabolic enzymes. No toxicologically relevant effects of PG/VG aerosols (up to 1.520 mg PG/L + 1.890 mg VG/L) were observed, and no adverse effects for PG/VG/nicotine were observed up to 438/544/6.6 mg/kg/day. This study demonstrates how complementary systems toxicology analyses can reveal, even in the absence of observable adverse effects, subtoxic and adaptive responses to pharmacologically active compounds such as nicotine.


Association of Hodgkin's lymphoma with Epstein Barr virus infection.
The role of Epstein Barr virus (EBV) in the onset of Hodgkin's lymphoma has been a subject of ongoing research. However, confirmation of EBV oncogenic involvement was not possible due to the small number of neoplastic cells characteristic for this type of tumor. Presence of EBV infection in neoplastic and non-neoplastic cells was analyzed in 81 cases of Hodgkin's lymphoma. In neoplastic cells, using an immunohistochemical method, latent membrane protein 1 (LMP1) was found in 33,3% of cases, while in situ hybridization results demonstrated the presence of EBER RNA in 48,1% of the cases. EBER RNA was found in non-neoplastic lymphocytes in 38,3% of cases. EBV is most frequently associated with Hodgkin's lymphoma in the first and seventh decade of life, specifically the nodular sclerosis subtype. No apparent difference was observed in the association of Hodgkin's lymphoma with EBV between genders, or in relation to clinical stage of the disease and average age of the patient. However, association with childhood age is significantly greater in comparison to adults. EBV associated disease shows a significantly greater prevalence in T lymphocytes. Slightly more abundant are cytotoxic T lymphocytes, which are also more frequently in contact with Reed-Sternberg cells, although there is no difference in number and positioning of histiocytes. Variations between the data on the association of EBV with Hodgkin's lymphoma among studies from different parts of the world suggest that factors of age, gender, ethnic background and social status might present biological modifiers of EBV influence on the pathogenesis of this neoplasm. The differences in non-neoplastic infiltrate EBV+ and EBV- lymphoma indicate the effect of the virus on the immune interaction of tumor and host in this disease.


NK1 receptors mediated release of 6-keto-PGF1 alpha from the ex vivo perfused canine ileum.
The purpose of this study was to determine the effects of tachykinins on prostanoid production by the dog ileum and to characterize the tachykinin receptor(s) responsible for the principal eicosanoid shown to be released, 6-keto-PGF1 alpha. Substance P, the selective NK1 receptor agonist [Sar9,Met(O2)11]substance P and neurokinin A caused concentration-dependent production of 6-keto-PGF1 alpha; neurokinin A was least potent. The selective NK2 agonist [Nle10]neurokinin A(4-10) had no effect. The selective NK1 antagonist CP-96,345 (10(-7) M), blocked 6-keto-PGF1 alpha release from substance P (10(-7) M), [Sar9,Met(O2)11]substance P (10(-7) M) and neurokinin A (10(-7) M). Although the putative NK2 antagonist MEN 10207 (10(-7) M) partially blocked the 6-keto-PGF1 alpha release induced by neurokinin A (10(-7) M), we conclude that all these peptides acted primarily on NK1 receptors to induce 6-keto-PGF1 alpha. Additional experiments suggest that a major site of production of 6-keto-PGF1 alpha in the canine ileum may be the vasculature, but these experiments do not exclude other sources such as intestinal muscle for this prostanoid. Calcium-free Krebs' solution partially reduced the release of 6-keto-PGF1 alpha to substance P (10(-7) M), implying that extracellular calcium helps support tachykinin-induced production of 6-keto-PGF1 alpha. Blockade of synthesis of another vasoactive mediator, endothelium-derived relaxing factor (nitric oxide), by N omega-L-arginine methyl ester) did not alter substance P-induced release of 6-keto-PGF1 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)


Substrates, inducers, inhibitors and structure-activity relationships of human Cytochrome P450 2C9 and implications in drug development.
Cytochrome P450 2C9 (CYP2C9) is one of the most abundant CYP enzymes in the human liver. CYP2C9 metabolizes more than 100 therapeutic drugs, including tolbutamide, glyburide, diclofenac, celecoxib, torasemide, phenytoin losartan, and S-warfarin). Some natural and herbal compounds are also metabolized by CYP2C9, probably leading to the formation of toxic metabolites. CYP2C9 also plays a role in the metabolism of several endogenous compounds such as steroids, melatonin, retinoids and arachidonic acid. Many CYP2C9 substrates are weak acids, but CYP2C9 also has the capacity to metabolise neutral, highly lipophilic compounds. A number of ligand-based and homology models of CYP2C9 have been reported and this has provided insights into the binding of ligands to the active site of CYP2C9. Data from the site-directed mutagenesis studies have revealed that a number of residues (e.g. Arg97, Phe110, Val113, Phe114, Arg144, Ser286, Asn289, Asp293 and Phe476) play an important role in ligand binding and determination of substrate specificity. The resolved crystal structures of CYP2C9 have confirmed the importance of these residues in substrate recognition and ligand orientation. CYP2C9 is activated by dapsone and its analogues and R-lansoprazole in a stereo-specific and substrate-dependent manner, probably through binding to the active site and inducing positive cooperativity. CYP2C9 is subject to induction by rifampin, phenobarbital, and dexamethasone, indicating the involvement of pregnane X receptor, constitutive androstane receptor and glucocorticoid receptor in the regulation of CYP2C9. A number of compounds have been found to inhibit CYP2C9 and this may provide an explanation for some clinically important drug interactions. Tienilic acid, suprofen and silybin are mechanism-based inhibitors of CYP2C9. Given the critical role of CYP2C9 in drug metabolism and the presence of polymorphisms, it is important to identify drug candidates as potential substrates, inducer or inhibitors of CYP2C9 in drug development and drug discovery scientists should develop drugs with minimal interactions with this enzyme. Further studies are warranted to explore the molecular determinants for ligand-CYP2C9 binding and the structure-activity relationships.


Reversible Electrochemical Charging of n-Type Conjugated Polymer Electrodes in Aqueous Electrolytes.
Conjugated polymers achieve redox activity in electrochemical devices by combining redox-active, electronically conducting backbones with ion-transporting side chains that can be tuned for different electrolytes. In aqueous electrolytes, redox activity can be accomplished by attaching hydrophilic side chains to the polymer backbone, which enables ionic transport and allows volumetric charging of polymer electrodes. While this approach has been beneficial for achieving fast electrochemical charging in aqueous solutions, little is known about the relationship between water uptake by the polymers during electrochemical charging and the stability and redox potentials of the electrodes, particularly for electron-transporting conjugated polymers. We find that excessive water uptake during the electrochemical charging of polymer electrodes harms the reversibility of electrochemical processes and results in irreversible swelling of the polymer. We show that small changes of the side chain composition can significantly increase the reversibility of the redox behavior of the materials in aqueous electrolytes, improving the capacity of the polymer by more than one order of magnitude. Finally, we show that tuning the local environment of the redox-active polymer by attaching hydrophilic side chains can help to reach high fractions of the theoretical capacity for single-phase electrodes in aqueous electrolytes. Our work shows the importance of chemical design strategies for achieving high electrochemical stability for conjugated polymers in aqueous electrolytes.


Mercuric chloride-induced cytotoxicity and compensatory hypertrophy in rat kidney proximal tubular cells.
Previous work showed that uninephrectomized (NPX) rats are more susceptible to the nephropathy induced by some doses of HgCl2 than sham-operated (SHAM) rats. The aim of the present study was to investigate the cytotoxic effects of HgCl2 in proximal tubular (PT) cells isolated from the kidney(s) of both NPX and SHAM rats. The study was designed to test if isolated PT cells that have undergone compensatory hypertrophy in vivo are more sensitive to the cytotoxic effects of HgCl2 in vitro than PT cells isolated from the kidneys of control animals. PT cells were purified from suspensions of renal cortical cells by Percoll density gradient centrifugation. The cellular content of protein (mg/10(6) cells) was 68% higher and the cellular specific activity (mU/10(6) cells) of lactate dehydrogenase was 56% higher in PT cells isolated from NPX rats than in PT cells isolated from SHAM rats. The cytotoxicity of HgCl2, as assessed by inhibition of lactate dehydrogenase activity, exhibited a steep concentration dependence. In the presence of bovine serum albumin, PT cells from both NPX and SHAM rats displayed no signs of cellular injury at concentrations of HgCl2 less than or equal to 0.1 mM. At concentrations of HgCl2 greater than or equal to 0.25 mM, cellular necrosis occurred rapidly in all PT cells. In the absence of bovine serum albumin, cellular injury and death occurred at concentrations of HgCl2 as low as 0.01 mM. PT cells from NPX rats exhibited signs of cellular injury at lower concentrations of HgCl2 than PT cells from SHAM rats when BSA was absent from the extracellular medium. Preincubation of PT cells from both NPX and SHAM rats with glutathione provided the cells with concentration-dependent protection from the cytotoxic effects of HgCl2. Preincubation of PT cells from both NPX and SHAM rats with the chelating agent 2,3-dimercapto-1-propane sulfonate provided cells with complete protection from the cytotoxic effects of HgCl2 when the concentration of 2,3-dimercapto-1-propane sulfonate was slightly below or higher than the concentration of HgCl2. Results of this study demonstrate the usefulness of freshly isolated PT cells from NPX rats as an in vitro model system to investigate biochemical mechanisms by which compensatory renal growth alters susceptibility to chemical-induced nephrotoxicity.


Myrciaria cauliflora extracts attenuate diabetic nephropathy involving the Ras signaling pathway in streptozotocin/nicotinamide mice on a high fat diet.
Diabetic nephropathy (DN) is a major cause of end-stage renal disease and its mortality is continuously increasing worldwide. Previous studies indicate that reactive oxygen species play an important role in high glucose-induced renal injury. Myrciaria cauliflora has been reported as a functional food rich in anthocyanins possessing anti-oxidative and anti-inflammatory properties. This study examined whether M. cauliflora extracts (MCE) can attenuate diabetic nephropathy progression in type 2 diabetes mellitus mice. First, the composition of the anthocyanins and polyphenols of MCE were determined by high-performance liquid chromatography and spectrophotometry. One hundred mg/kg of streptozotocin and 240 mg/kg nicotinamide were administered to C57BL/6J mice fed a high fat diet and varied concentrations of MCE. The plasma glucose concentration, body weight, oral glucose tolerance, blood pressure, renal ultrasound ultrasonic wave were monitored every 2 weeks. Following euthanasia, the kidneys of the mice were analyzed using hematoxylin-eosin, periodic acid Schiff, Masson's trichrome, and immunohistochemistry staining. The results showed that MCE stabilized the plasma glucose and indirectly improved insulin sensitivity in diabetic mice. In addition, diabetes-caused glomerular atrophy, accumulation of saccharide, and formation of collagen IV were recovered or reduced under treatment with MCE in diabetic mice. Our results indicate that MCE has beneficial effects in DN and the mechanism has been confirmed to inhibit Ras/PI3K/Akt and kidney fibrosis related proteins. This work illustrates the potential of MCE rich in anthocyanins and polyphenols as a natural food to inhibit DN.


Obesity prevention and the role of hospital and community-based health services: a scoping review.
Control of obesity is an important priority to reduce the burden of chronic disease. Clinical guidelines focus on the role of primary healthcare in obesity prevention. The purpose of this scoping review is to examine what the published literature indicates about the role of hospital and community based health services in adult obesity prevention in order to map the evidence and identify gaps in existing research. Databases were searched for articles published in English between 2006 and 2016 and screened against inclusion and exclusion criteria. Further papers were highlighted through a manual search of the reference lists. Included papers evaluated interventions aimed at preventing overweight and obesity in adults that were implemented within and/or by hospital and community health services; were an empirical description of obesity prevention within a health setting or reported health staff perceptions of obesity and obesity prevention. The evidence supports screening for obesity of all healthcare patients, combined with referral to appropriate intervention services but indicates that health professionals do not typically adopt this practice. As well as practical issues such as time and resourcing, implementation is impacted by health professionals' views about the causes of obesity and doubts about the benefits of the health sector intervening once someone is already obese. As well as lacking confidence or knowledge about how to integrate prevention into clinical care, health professional judgements about who might benefit from prevention and negative views about effectiveness of prevention hinder the implementation of practice guidelines. This is compounded by an often prevailing view that preventing obesity is a matter of personal responsibility and choice. This review highlights that whilst a population health approach is important to address the complexity of obesity, it is important that the remit of health services is extended beyond medical treatment to incorporate obesity prevention through screening and referral. Further research into the role of health services in obesity prevention should take a systems approach to examine how health service structures, policy and practice interrelationships, and service delivery boundaries, processes and perspectives impact on changing models of care.


Establishing hospital-based trauma registry systems: lessons from Kenya.
In the developing world, data about the burden of injury, injury outcomes, and complications of care are limited. Hospital-based trauma registries are a data source that can help define this burden. Under the trauma care component of the Bloomberg Global Road Safety Partnership, trauma registries have been implemented at three sites in Kenya. We describe the challenges and lessons learned from this effort. A paper-based trauma surveillance form was developed, in collaboration with local hospital partners, to collect data on all trauma patients presenting for care. The form includes demographic information, pre-hospital care given, and patient care and clinical information necessary to calculate estimated injury surveillance. The type of data collected was standardized across all three sites. Frequent reviews of the data collection process, quality, and completeness, in addition to regular meetings and conference calls, have allowed us to optimize the process to improve efficiency and make corrective actions where required. Trauma registries have been implemented in three hospitals in Kenya, with potential for expansion to other hospitals and facilities caring for injured patients. The process of establishing registries was associated with both general and site-specific challenges. Problems were identified in planning, data collection, entry processes, and analysis. Problems were addressed when identified, resulting in improved data quality. Trauma registries are a key data source for defining the burden of injury and developing quality improvement processes. Trauma registries were implemented at three sites in Kenya. Problems and challenges in data collection were identified and corrected. Through the registry data, gaps in care were identified and systemic changes made to improve the care of the injured.


Peripheral blood cell telomere length measurements indicate that hematopoietic stem cell turnover is not significantly increased in whole blood and apheresis PLT donors.
The telomere length (TEL) of peripheral blood leukocytes (PBLs) can be used to estimate hematopoietic stem cell turnover. The current study investigated whether the repetitive stimulation of the hematopoietic system caused by regular whole blood (WB) and PLT donations would affect PBL TEL. PBLs were obtained from healthy donors (n=94) with a history of at least 3 years of WB donation (median, 7.7 years; range, 3.0-43.0 years) plus additional apheresis PLT donations. The median (range) numbers of WB and PLT donations were 22.0 (6.0-194.0) and 42.0 (7.0-336.0), respectively. Additionally, samples were obtained from healthy nondonors (n=47). PBL TEL was measured with fluorescence in situ hybridization and flow cytometry (flow-FISH). Flow-FISH results were expressed in molecular equivalents of soluble fluorochrome units (MESF; 1000 MESF=1 kMESF) either as absolute (TEL) or as age-adjusted TEL (DeltaTEL). Donor granulocyte and lymphocyte TELs were 12.6 +/- 0.3 (mean +/- SEM) and 13.2 +/- 0.3 kMESF, respectively. No differences were observed when compared with corresponding nondonor data (granulocytes, 12.5 +/- 0.4 kMESF; lymphocytes, 13.6 +/- 0.5 kMESF). Furthermore, DeltaTEL values did not differ between the two groups and were not different from previously established reference values. In addition, neither donor data for age-adjusted TEL for granulocytes nor DeltaTEL for lymphocytes were correlated with either total years or total numbers of WB and/or PLT donations. Long-term WB and PLT donation does not affect PBW TEL as measured by flow-FISH, arguing against a significantly increased stem cell turnover.


Changes in the phosphoproteome of brown adipose tissue during hibernation in the ground squirrel, Ictidomys tridecemlineatus.
Mammalian hibernation is characterized by metabolic rate depression and a strong decrease in core body temperature that together create energy savings such that most species do not have to eat over the winter months. Brown adipose tissue (BAT), a thermogenic tissue that uses uncoupled mitochondrial respiration to generate heat instead of ATP, plays a major role in rewarming from deep torpor. In the present study we developed a label-free liquid chromatography mass spectrometry (LC-MS) strategy to investigate both differential protein expression and protein phosphorylation in BAT extracts from euthermic vs. hibernating ground squirrels (Ictidomys tridecemlineatus). In particular, we incorporated the filter-assisted sample preparation protocol, which provides a more in-depth analysis compared with gel-based and other LC-MS proteomics approaches. Surprisingly, mitochondrial membrane and matrix protein expression in BAT was largely constant between active euthermic squirrels and their hibernating counterparts. Validation by immunoblotting confirmed that the protein levels of mitochondrial respiratory chain complexes were largely unchanged in hibernating vs. euthermic animals. On the other hand, phosphoproteomics revealed that pyruvate dehydrogenase (PDH) phosphorylation increased during squirrel hibernation, confirmed by immunoblotting with phospho-specific antibodies. PDH phosphorylation leads to its inactivation, which suggests that BAT carbohydrate oxidation is inhibited during hibernation. Phosphorylation of hormone-sensitive lipase (HSL) was also found to increase during hibernation, suggesting that HSL would be active in BAT to produce the fatty acids that are likely the primary fuel for thermogenesis upon arousal. Increased perilipin phosphorylation along with that of a number of other proteins was also revealed, emphasizing the importance of protein phosphorylation as a regulatory mechanism during mammalian hibernation.