Ticagrelor: from discovery to Phase III clinical trial.
Ticagrelor (AZD6140), a cyclopentyl-triazolo-pyrimidine, is the first orally available antagonist of the ADP receptor of the P2Y12 subtype. Ticagrelor inhibits platelets in a reversible manner and does not require hepatic bioactivation. The pharmacology of ticagrelor indicates that it provides more consistent, more rapid and more potent platelet inhibition than clopidogrel. Preclinical and clinical studies with ticagrelor have demonstrated that this drug has excellent oral bioavailability. The Phase III clinical study of Platelet Inhibition and Patient Outcomes (PLATO) has shown that ticagrelor reduced ischemic events and all-cause mortality without an increase in major bleeding complications. Potential advantages of ticagrelor include more flexibility in its use if rapid onset of action is needed before percutaneous coronary interventions or when cessation is required before coronary artery bypass graft surgery. Potential disadvantages include more side effects such as dyspnea, ventricular pauses or an increase in concentrations of uric acid and creatinine. However, ticagrelor did not only reduce death due to vascular causes but also all-cause mortality. Further clinical trials in indications other than acute coronary syndrome are awaited.


SUN proteins and nuclear envelope spacing.
The nuclear envelope consists of 2 membranes separated by 30-50 nm, but how the 2 membranes are evenly spaced has been an open question in the field. Nuclear envelope bridges composed of inner nuclear membrane SUN proteins and outer nuclear membrane KASH proteins have been proposed to set and regulate nuclear envelope spacing. We tested this hypothesis directly by examining nuclear envelope spacing in Caenorhabditis elegans animals lacking UNC-84, the sole somatic SUN protein. SUN/KASH bridges are not required to maintain even nuclear envelope spacing in most tissues. However, UNC-84 is required for even spacing in body wall muscle nuclei. Shortening UNC-84 by 300 amino acids did not narrow the nuclear envelope space. While SUN proteins may play a role in maintaining nuclear envelope spacing in cells experiencing forces, our data suggest they are dispensable in most cells.


Sensitivity of the Speech Intelligibility Index to the Assumed Dynamic Range.
This study aims to evaluate the sensitivity of the speech intelligibility index (SII) to the assumed speech dynamic range (DR) in different languages and with different types of stimuli. Intelligibility prediction uses the absolute transfer function (ATF) to map the SII value to the predicted intelligibility for a given stimuli. To evaluate the sensitivity of the predicted intelligibility to the assumed DR, ATF-transformed SII scores for English (words), Korean (sentences), and Mandarin (sentences) were derived for DRs ranging from 10 dB to 60 dB. Increasing the assumed DR caused steeper ATFs for all languages. However, high correlation coefficients between predicted and measured intelligibility scores were observed for DRs from 20 dB to 60 dB for ATFs in English, Korean, and Mandarin. Results of the present study indicate that the intelligibility computed from the SII is not sensitive to the assumed DR. The 30-dB DR commonly used in computing the SII is thus a reasonable assumption that produces accurate predictions for different languages and different types of stimuli.


Lateral Gene Transfer Dynamics in the Ancient Bacterial Genus Streptomyces.
Lateral gene transfer (LGT) profoundly shapes the evolution of bacterial lineages. LGT across disparate phylogenetic groups and genome content diversity between related organisms suggest a model of bacterial evolution that views LGT as rampant and promiscuous. It has even driven the argument that species concepts and tree-based phylogenetics cannot be applied to bacteria. Here, we show that acquisition and retention of genes through LGT are surprisingly rare in the ubiquitous and biomedically important bacterial genus Streptomyces Using a molecular clock, we estimate that the Streptomyces bacteria are ~380 million years old, indicating that this bacterial genus is as ancient as land vertebrates. Calibrating LGT rate to this geologic time span, we find that on average only 10 genes per million years were acquired and subsequently maintained. Over that same time span, Streptomyces accumulated thousands of point mutations. By explicitly incorporating evolutionary timescale into our analyses, we provide a dramatically different view on the dynamics of LGT and its impact on bacterial evolution.IMPORTANCE Tree-based phylogenetics and the use of species as units of diversity lie at the foundation of modern biology. In bacteria, these pillars of evolutionary theory have been called into question due to the observation of thousands of lateral gene transfer (LGT) events within and between lineages. Here, we show that acquisition and retention of genes through LGT are exceedingly rare in the bacterial genus Streptomyces, with merely one gene acquired in Streptomyces lineages every 100,000 years. These findings stand in contrast to the current assumption of rampant genetic exchange, which has become the dominant hypothesis used to explain bacterial diversity. Our results support a more nuanced understanding of genetic exchange, with LGT impacting evolution over short timescales but playing a significant role over long timescales. Deeper understanding of LGT provides new insight into the evolutionary history of life on Earth, as the vast majority of this history is microbial.


Production of the Plant Hormone Auxin by Salmonella and Its Role in the Interactions with Plants and Animals.
The ability of human enteric pathogens to colonize plants and use them as alternate hosts is now well established. Salmonella, similarly to phytobacteria, appears to be capable of producing the plant hormone auxin via an indole-3-pyruvate decarboxylase (IpdC), a key enzyme of the IPyA pathway. A deletion of the Salmonella ipdC significantly reduced auxin synthesis in laboratory culture. The Salmonella ipdC gene was expressed on root surfaces of Medicago truncatula. M. truncatula auxin-responsive GH3::GUS reporter was activated by the wild type Salmonella, and not but the ipdC mutant, implying that the bacterially produced IAA (Indole Acetic Acid) was detected by the seedlings. Seedling infections with the wild type Salmonella caused an increase in secondary root formation, which was not observed in the ipdC mutant. The wild type Salmonella cells were detected as aggregates at the sites of lateral root emergence, whereas the ipdC mutant cells were evenly distributed in the rhizosphere. However, both strains appeared to colonize seedlings well in growth pouch experiments. The ipdC mutant was also less virulent in a murine model of infection. When mice were infected by oral gavage, the ipdC mutant was as proficient as the wild type strain in colonization of the intestine, but it was defective in the ability to cross the intestinal barrier. Fewer cells of the ipdC mutant, compared with the wild type strain, were detected in Peyer's patches, spleen and in the liver. Orthologs of ipdC are found in all Salmonella genomes and are distributed among many animal pathogens and plant-associated bacteria of the Enterobacteriaceae, suggesting a broad ecological role of the IpdC-catalyzed pathway.


Plant-specific microtubule-associated protein SPIRAL2 is required for anisotropic growth in Arabidopsis.
In diffusely growing plant cells, cortical microtubules play an important role in regulating the direction of cell expansion. Arabidopsis (Arabidopsis thaliana) spiral2 (spr2) mutant is defective in directional cell elongation and exhibits right-handed helical growth in longitudinally expanding organs such as root, hypocotyl, stem, petiole, and petal. The growth of spr2 roots is more sensitive to microtubule-interacting drugs than is wild-type root growth. The SPR2 gene encodes a plant-specific 94-kD protein containing HEAT-repeat motifs that are implicated in protein-protein interaction. When expressed constitutively, SPR2-green fluorescent protein fusion protein complemented the spr2 mutant phenotype and was localized to cortical microtubules as well as other mitotic microtubule arrays in transgenic plants. Recombinant SPR2 protein directly bound to taxol-stabilized microtubules in vitro. Furthermore, SPR2-specific antibody and mass spectrometry identified a tobacco (Nicotiana tabacum) SPR2 homolog in highly purified microtubule-associated protein fractions from tobacco BY-2 cell cultures. These results suggest that SPR2 is a novel microtubule-associated protein and is required for proper microtubule function involved in anisotropic growth.


Systemic short chain fatty acids limit antitumor effect of CTLA-4 blockade in hosts with cancer.
Gut microbiota composition influences the clinical benefit of immune checkpoints in patients with advanced cancer but mechanisms underlying this relationship remain unclear. Molecular mechanism whereby gut microbiota influences immune responses is mainly assigned to gut microbial metabolites. Short-chain fatty acids (SCFA) are produced in large amounts in the colon through bacterial fermentation of dietary fiber. We evaluate in mice and in patients treated with anti-CTLA-4 blocking mAbs whether SCFA levels is related to clinical outcome. High blood butyrate and propionate levels are associated with resistance to CTLA-4 blockade and higher proportion of Treg cells. In mice, butyrate restrains anti-CTLA-4-induced up-regulation of CD80/CD86 on dendritic cells and ICOS on T cells, accumulation of tumor-specific T cells and memory T cells. In patients, high blood butyrate levels moderate ipilimumab-induced accumulation of memory and ICOS + CD4 + T cells and IL-2 impregnation. Altogether, these results suggest that SCFA limits anti-CTLA-4 activity.


Identification of spinal neurons in the embryonic and larval zebrafish.
Previous studies indicated that the developing fish spinal cord was a simple system containing a small number of distinguishable neuronal cell types (Eisen et al., Nature 320:269-271, '86; Kuwada, Science, 233:740-746, '86). To verify this we have characterized the cellular anatomy of the spinal cord of developing zebrafish in order to determine the number, identities, and organization of the spinal neurons. Spinal neurons were labeled by intracellular dye injections, application of an axonal tracer dye to all or subsets of the axonal tracts, and application of antibodies which recognize embryonic neurons. We found that nine classes of neurons could be identified based on soma size and position, pattern of dendrites, axonal trajectory, and time of axonogenesis. These are two classes of axial motor neurons, which have been previously characterized (Myers, J. Comp. Neurol. 236:555-561, '85), one class of sensory neurons, and six classes of interneurons. One of the interneuron classes could be subclassified as primary and secondary based on criteria similar to those used to classify the axial motor neurons into primary and secondary classes. The early cord (18-20 hours) is an extremely simple system and contains approximately 18 lateral cell bodies per hemisegment, which presumably are post-mitotic cells. By this stage, five of the neuronal classes have begun axonogenesis including the primary motor neurons, sensory neurons, and three classes of interneurons. By concentrating on these early stages when the cord is at its simplest, pathfinding by growth cones of known identities can be described in detail. Then it should be possible to test many different mechanisms which may guide growth cones in the vertebrate central nervous system (CNS).


Stoichiometric and catalytic scavengers as protection against nerve agent toxicity: a mini review.
Currently fielded treatments for nerve agent intoxication promote survival, but do not afford complete protection against either nerve agent-induced motor and cognitive deficits or neuronal pathology. The use of human plasma-derived butyrylcholinesterase (HuBuChE) to neutralize the toxic effects of nerve agents in vivo has been shown to both aid survival and protect against decreased cognitive function after nerve agent exposure. Recently, a commercially produced recombinant form of human butyrylcholinesterase (r-HuBuChE; PharmAthene Inc.) expressed in the milk of transgenic goats has become available. This material is biochemically similar to plasma-derived HuBuChE in in vitro assays. The pharmacokinetic characteristics of a polyethylene glycol coated (pegylated) form of r-HuBuChE were determined in guinea pigs; the enzyme was rapidly bioavailable with a half-life (t(1/2)) and pharmacokinetic profile that resembled that of plasma-derived huBuChE. Guinea pigs were injected with 140mg/kg (i.m.) of pegylated r-HuBuChE 18h prior to exposure (sc) to 5.5xLD(50) VX or soman. VX and soman were administered in a series of three injections of 1.5xLD(50), 2.0xLD(50), and 2.0xLD(50), respectively, with injections separated by 2h. Pretreatment with pegylated r-HuBuChE provided 100% survival against multiple lethal doses of VX and soman. Guinea pigs displayed no signs of nerve agent toxicity following exposure. Assessments of motor activity, coordination, and acquisition of spatial memory were performed for 2 weeks following nerve agent exposure. There were no measurable decreases in motor or cognitive function during this period. In contrast, animals receiving 1.5xLD(50) challenges of soman or VX and treated with standard atropine, 2-PAM, and diazepam therapy showed 50 and 100% survival, respectively, but exhibited marked decrements in motor function and, in the case of GD, impaired spatial memory acquisition. The advances in this field have resulted in the decision to select both the plasma-derived and the recombinant form of BuChE for advanced development and transition to clinical trials. Efforts have now been expanded to identify a catalytic protein capable of not only binding, but also rapidly hydrolyzing the standard threat nerve agents. Recent work has focused on paraoxonase-1 (PON1), a naturally occurring human serum enzyme with the capacity to catalyze the hydrolysis of nerve agents, albeit too slowly to afford dramatic protection. Using rational design, several amino acids involved in substrate binding have been identified and site-directed mutations have revealed that residue H115 plays an important role in binding. In addition, the stereospecificity of PON1 for the catalytic hydrolysis of soman has been examined. The enzyme exhibits a slight stereospecificity for the C+P+ isomer of soman, which is due more to preferential binding than to selective hydrolysis of this isomer. The results suggest that it may be possible to engineer a mutant form of PON1 with enhanced activity and stereospecificity for the most toxic nerve agent isoforms.


Testing Separability and Independence of Perceptual Dimensions with General Recognition Theory: A Tutorial and New R Package (grtools).
Determining whether perceptual properties are processed independently is an important goal in perceptual science, and tools to test independence should be widely available to experimental researchers. The best analytical tools to test for perceptual independence are provided by General Recognition Theory (GRT), a multidimensional extension of signal detection theory. Unfortunately, there is currently a lack of software implementing GRT analyses that is ready-to-use by experimental psychologists and neuroscientists with little training in computational modeling. This paper presents grtools, an R package developed with the explicit aim of providing experimentalists with the ability to perform full GRT analyses using only a couple of command lines. We describe the software and provide a practical tutorial on how to perform each of the analyses available in grtools. We also provide advice to researchers on best practices for experimental design and interpretation of results when applying GRT and grtools.


p53 Codon 72 polymorphism and the risk of esophageal squamous cell carcinoma.
The polymorphisms of the tumor suppressor gene p53 have been extensively investigated in numerous malignant tumors, particularly carcinomas associated with human papillomavirus (HPV) infection. However, the results remain controversial. To address a potential correlation between the p53 genotypes and the risk of esophageal squamous cell carcinoma (ESCC), we investigated the p53 codon 72 polymorphism in 435 patients with ESCC and 550 cancer-free subjects from the same geographical region. p53 Arg/Arg genotype was significantly increased in ESCC cases compared with control subjects (85.7 vs. 49.6%, P < 0.001), resulting in an elevated ESCC risk (OR = 6.48, 95% CI = 4.65-9.03). In addition, among p53 Arg/Arg carriers, HPV infection, smoking, and drinking might further increase the risk of ESCC development.


Effectiveness of allogeneic CD3AK cells on transplanted human renal cell cancer in mice with severe combined immune deficiency.
To assess the activity of allogeneic anti-CD3 antibody induced activated killer (CD3AK) cells on transplanted human renal cell cancer (RCC) in mice with severe combined immune deficiency (SCID), thus to provide theoretical and experimental support for clinical application of allogeneic CD3AK cells in the treatment of RCC. A culture system which can massively increase allogeneic CD3AK cells was constructed. CCK-8 method was used to detect lethal effect of allogeneic CD3AK cells on human OS-RC-2 renal cancer cell line. Then, tumor-bearing mice models were constructed. SCID mice were randomly divided into four groups: group A (caudal vein was injected with allogeneic CD3AK cells before tumor bearing), group B (the control group of group A: caudal vein was injected with PBS before tumor bearing), group C (caudal vein was injected with allogeneic CD3AK cells after tumor bearing) and group D (the control group of group C: caudal vein was injected with PBS after tumor bearing), and spleen parameters were calculated to observe any inhibitory effect of allogeneic CD3AK cells on the growth of renal cancer cells, as well as their effect on the immune system of mice. Compared with the control groups B and D, spleen parameters of groups A and C increased significantly (p<0.05); compared with group C, spleen parameters of group A showed no obvious difference (p>0.05); compared with the control groups B and D, tumor weight of groups A and C decreased significantly and tumors grew slowly (p<0.05); the weight of mice in all groups increased while the weight of mice in the experimental groups increased less than the weight of mice in the control groups (p<0.05); compared with group C, tumor weight of group A reduced and grew slowly (p<0.05). Immunohistochemistry revealed that CD3+ cells showed positive expression in tumor tissues of mice in group C, while no positive expression was found in tumor tissues of mice in other groups. Allogeneic CD3AK cells exerted lethal effect on OS-RC-2 cell line and significant inhibitory activity on the growth of transplanted cancer in mice with SCID. Also CD3AK cells expressed certain preventive effect on the development of implanted cancer in SCID mice; allogeneic CD3AK cells possessed antitumor activity and could enhance the immunologic functions of SCID mice with human renal cell-bearing cancer.


Photoacoustic/Fluorescence Dual-Modality Probe for Biothiol Discrimination and Tumor Diagnosis in Cells and Mice.
Developing probes to simultaneously detect and discriminate biothiols is important, yet challenging. Activatable photoacoustic (PA) probes for discriminating biothiols in vivo are still lacking, and this hinders the diagnosis of thiol-related diseases. Herein we present the first PA and fluorescence dual-modality probe MB-NBD for discriminating different biothiol species. The probe has the advantages of both fluorescence imaging and PA imaging (high sensitivity and deep penetration) with distinct signal patterns toward hydrogen sulfide (H2S), cysteine/homocysteine (Cys/Hcy), and glutathione (GSH) treatment. The biothiol-activated product of MB-NBD exhibits enhancements in near-infrared fluorescence (NIRF) at 690 nm and absorbance/PA at 664 nm upon fast reaction, allowing it to selectively detect biothiol species over other reactive species. On the other hand, MB-NBD displays characteristic absorbance enhancement at 547 nm toward H2S, rendering specific detection of H2S. In addition, the specific enhancements in absorbance/PA at 470 nm and fluorescence at 550 nm toward Cys/Hcy treatment endows the probe with the capability of selectively detecting Cys/Hcy. Furthermore, MB-NBD is able to discriminate Cys and GSH by fluorescent imaging in live-cell and ratiometric PA imaging in mice experiments. MB-NBD has been successfully used to diagnose tumors by dual-channel ratiometric PA imaging.


Using phenolic polymers to control the size and morphology of calcium carbonate microparticles.
Calcium carbonate (CaCO3) is a naturally occurring mineral that occurs in biology and is used industrially. Due to its benign nature, CaCO3 microparticles have found use in the food and medical fields, where the specific size of the microparticles determine their functionality and potential applications. We demonstrate that phenolic polymers with different numbers of hydroxy groups can be used to control the diameter of CaCO3 microparticles in a range of 2-9 μm, and obtained particles were relatively uniform. The largest particles (∼9 μm in diameter) were obtained using poly(2,3,4,5-tetrahydroxystyrene) (P4HS), which showed the highest water solubility among the tested phenolic polymers. The polymer concentration and stirring speed influenced the size of microparticles, where the size of the obtained particles became smaller as the concentrations of phenolic polymers increased and as the stirring speed increased, both likely due to promoting the formation of a large number of individual crystal seeds by shielding seed-seed fusion and increasing the chances for precursor contact, respectively. The preparation time and temperature had a great influence on the morphology of the CaCO3 particles, where vaterite transforms into calcite over time. Specifically, aragonite crystals were observed at preparation temperature of 80 °C and vaterite particles with rough surfaces were obtained at 40 °C. Molecular weight and scale of reaction were also factors which affect the size and morphologies of CaCO3 particles. This research represents a facile method for producing relatively monodisperse CaCO3 microparticles with diameters that have previously proven difficult to access.


Blood flow switching among pulmonary capillaries is decreased during high hematocrit.
Pulmonary capillary perfusion within a single alveolar wall continually switches among segments, even when large-vessel hemodynamics are constant. The mechanism is unknown. We hypothesize that the continually varying size of plasma gaps between individual red blood cells affects the likelihood of capillary segment closure and the probability of cells changing directions at the next capillary junction. We assumed that an increase in hematocrit would decrease the average distance between red blood cells, thereby decreasing the switching at each capillary junction. To test this idea, we observed 26 individual alveolar capillary networks by using videomicroscopy of excised canine lung lobes that were perfused first at normal hematocrit (31-43%) and then at increased hematocrit (51-62%). The number of switches decreased by 38% during increased hematocrit (P < 0.01). These results support the idea that a substantial part of flow switching among pulmonary capillaries is caused by the particulate nature of blood passing through a complex network of tubes with continuously varying hematocrit.


[Results of irradiation using different radiation sources in the treatment of malignant parotid tumors].
Patients with malignant parotid tumors were treated at two different centres with slightly ionizing radiation (110 patients) and neutron therapy (15 patients). The treatment results are compared. The principles of comparison applied in this analysis are identical. The histologic classification was made according to the W.H.O. recommendation of 1972 and the staging according to Becske. Local tumor control is the criterion applied in the comparative assessment of both methods. The analysis has shown that there is no significant difference in the treatment results within the individual stages of tumor's advancement after application of an orthovolt therapy and a telecobalt therapy. Patients treated in early stages (stages I and III) had a substantially higher rate of local tumor control (72%) than patients in advanced stages (46%, stages III and IV). A more favorable treatment results was achieved by neutron therapy in all stages of tumor's advancement. In this case the rate of patients with local tumor control (87%) was significantly higher than after radiotherapy with slightly ionizing radiation (46%).


Essential role of MD-2 in TLR4-dependent signaling during Helicobacter pylori-associated gastritis.
TLR4, a member of pattern recognition receptors, is the main receptor of LPS. MD-2 physically associates with TLR4 on the cell surface and confers LPS responsiveness. Helicobacter pylori LPS is one of the major virulence factors for induction of gastritis. We demonstrated in this study the role of MD-2 in TLR4-dependent signaling in H. pylori-associated gastritis. Gastric biopsy samples collected from patients with and without H. pylori infection and four gastric cancer cell lines were used for this study. TLR-4 and MD-2 expression in biopsy specimens and the cell lines was examined by using RT-PCR. Localization of TLR-4 in histological sections was evaluated by immunohistochemistry. For in vitro functional assays, we established stable transfectants of AGS cells expressing TLR4 and MD-2. Cellular distribution of TLR4 was examined by flow cytometry. NF-kappaB activation and activation of IL-8 and MD-2 promoters were assessed by reporter gene assay. H. pylori infection up-regulated the TLR4 and MD-2 expression in gastric mucosa. TLR4 staining was observed predominantly in epithelial cells, located in both the cytoplasm and at the apical surface. MD-2 transfection in AGS cells markedly increased cell surface expression of TLR4 and augmented the activation of NF-kappaB and IL-8 promoter upon stimulation with H. pylori LPS. Live H. pylori also stimulated transcriptional activation of MD-2. This study revealed that MD-2 expression is elevated in gastric epithelial cells during H. pylori infection, suggesting that the TLR4/MD-2 system is a potent receptor complex involved in the response to H. pylori LPS in the stomach.


Hospital Consumer Assessment of Healthcare Providers and Systems: Do Patient Demographics Affect Outcomes in Total Hip Arthroplasty?
The Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) score is a nationally standardized measure of a patient's hospital experience. This study aims to assess whether HCAHPS scores vary by demographic or surgical factors in patients undergoing primary total hip arthroplasty. Patients who completed an HCAHPS survey after a primary total hip arthroplasty between October 2011 and November 2016 were included in this study. Patient demographics and surgical factors were evaluated for correlations with individual HCAHPS questions. One thousand three hundred eighty-three HCAHPS questionnaires were reviewed for this study. Patients with a submitted HCAHPS response had an average age of 63.83 ± 10.17 years. Gender distribution was biased toward females at 57.27% (792 females) versus 42.73% (591 males). The average body mass index (BMI) was 28.68 ± 5.86 kg/m2. Race distribution was predominantly Caucasian at 81.49% (1127 patients), followed by "unknown" at 8.60% (119 patients) and African-American at 8.46% (117 patients). Home discharge occurred for 93.06% (1287 patients) versus 6.94% for facility discharge (96 patients). Mean length of stay was 2.41 ± 1.17 days. Each 1-year increase in age was positively correlated with a 0.16% increase in top-box response rate (β = 0.0016 ± 0.0008; P < .05). Male gender was correlated with a 4.61% increase in top-box response rate when compared to female gender (β = 0.0461 ± 0.0118; P < .01). BMI was found to be correlated with a 0.20% increase in HCAHPS response rates for each 1 kg/m2 increase (β = 0.0020 ± 0.0010; P < .05). For each day increase in length of stay, HCAHPS top-box response rates decrease by 3.41% (β = -0.0341 ± 0.0051; P < .0001). Race, marital status, smoking status, insurance type, and discharge disposition were not found to be significantly correlated with HCAHPS top-box response rate (P > .05). The HCAHPS quality measurement metric affects physician reimbursement and may be biased by a number of variables including sex, length of stay, and BMI, rather than a true reflection of the quality of their hospital experience. Further research is warranted to determine whether HCAHPS scores are an appropriate measure of the quality of care received.


Acute hemorrhagic pancreatic necrosis in mice. Effects of proteinase inhibitors on its induction.
An acute hemorrhagic pancreatitis with fat necrosis (AHPN) was induced in female mice fed a choline-deficient diet containing 0.5% DL-ethionine. The effect of various proteinase inhibitors on the induction of the pancreatitis was evaluated using three parameters, the mortality of the animals, the appearance before death of a shock-like state, and the severity of the pancreatic pathology. Treatment of the animals with leupeptin, pepstatin, chymostatin and/or antipain, proteinase inhibitors of microbial origin, resulted in a distinct attenuation of the severity of the induced process, whereas aprotinin and chloroquine had no effect. The results indicate that use of the microbial proteinase inhibitors should be considered as potential therapeutic agents in cases of pancreatitis.


The fermentation, isolation and characterization of macromolecular peptide antibiotics: AN-7A, -7B and -7D.
A group of new macromolecular peptide antibiotics, named AN-7, was isolated from the culture broth of Streptomyces griseoincarnatus AJ9424. AN-7 was fractionated into three different components, A, B and D. From 18 liters of culture broth (78 units/ml), 10 mg of AN-7A with a specific activity of 2,053 units/mg, 9 mg of AN-7B (1,167 units/mg) and 11 mg of AN-7D (6,225 units/mg) were obtained. All of these samples gave single bands on polyacrylamide gel electrophoresis. They are acidic polypeptides with molecular weights ranging from 12,400 to 13,000. Their UV absorption spectra showed maxima peaks at 280 nm and shoulders at 290 nm. Each AN-7 component has a nonprotein chromophoric component. AN-7A, -7B and -7D have no antibacterial activity against the Gram-negative bacteria tested but strongly inhibit the growth of Gram-positive bacteria and Escherichia coli MP2, a macromolecule permeable mutant strain. The AN-7 components are mutagenic. These antibiotics inhibit the in vitro growth of L1210 cells (ED50 0.13 approximately 0.18 micrograms/ml). AN-7A, -7B and -7D also inhibit the growth of L1210 cells in mice.


RPE65 operates in the vertebrate visual cycle by stereospecifically binding all-trans-retinyl esters.
RPE65 is a major protein of unknown function found associated with the retinyl pigment epithelial (RPE) membranes [Hamel, C. P., Tsilou, E., Pfeffer, B. A., Hooks, J. J., Detrick, B., and Redmond, T. M. (1993) J. Biol. Chem. 268, 15751-15757; Bavik, C. O., Levy, F., Hellman, U., Wernstedt, C., and Eriksson, U. (1993) J. Biol. Chem. 268, 20540-20546]. RPE65 knockouts fail to synthesize 11-cis-retinal, the chromophore of rhodopsin, and accumulate all-trans-retinyl esters in the RPE. Previous studies have also shown that RPE65 is specifically labeled with all-trans-retinyl ester based affinity labeling agents, suggesting a retinyl ester binding role for the protein. In the present work, we show that purified RPE65 binds all-trans-retinyl palmitate (tRP) with a K(D) = 20 pM. These quantitative experiments are performed by measuring the quenching of RPE65 fluorescence by added tRP. The binding for tRP is highly specific because 11-cis-retinyl palmitate binds with a K(D) = 14 nM, 11-cis-retinol binds with a K(D) = 3.8 nM, and all-trans-retinol (vitamin A) binds with a K(D) = 10.8 nM. This stereospecificity for tRP is to be compared to the binding of retinoids to BSA, where virtually no discrimination is found in the binding of the same retinoids. This work provides further evidence that RPE65 functions by binding to and mobilizing the highly hydrophobic all-trans-retinyl esters, allowing them to enter the visual cycle.


Multi-frequency bioelectrical impedance analysis (BIA) compared to magnetic resonance imaging (MRI) for estimation of fat-free mass in colorectal cancer patients treated with chemotherapy.
Changes in body composition in cancer patients during chemotherapy are associated with treatment related toxicities or mortalities. Thus, it is relevant to identify accessible, relatively inexpensive, portable and reliable tools for evaluation of body composition in cancer patients during the course of their treatments. To examine relationships between single cross-sectional thighs magnetic resonance imaging (MRI), skeletal muscle mass (SM) as reference and multi-frequency bioelectrical impedance analysis (BIA) fat free mass (FFM) in patients with colorectal cancer undergoing chemotherapy. In an observational, prospective study we examine the relationships between single cross-sectional thighs MRI (T1-weighted (1.5 T) SM compared to FFM BIA (8-electrodes multi-frequency Tanita MC780MA)) and FFM skin-fold thickness (ST) (4-points (Harpenden, Skinfold Caliper)) and SM equation for non-obese persons from Lee et al. 2000 (L2000) (based on age, height, weight, sex and race). FFM and SM (kg) were calculated based on either area (MRI) or weight. 18 CRC patients (10 males and 8 females) with mean (SD) age 67 yr (6) were measured at baseline, and 13 were available for follow-up. BIA overestimated FFM kg for all 31 measurements with mean (SD) 18.0 kg (6.0) compared to the MRI. ST overestimated FFM kg with mean 12.4 kg (6.2) and L2000 underestimated SM kg in 18 measurements and overestimated in 13 with a total mean of -4.3 kg (6.8). BIA and ST were the best alternatives to MRI as they showed constant and thereby correctable errors. The equation, L2000, carried the smallest average measurement error but it was non-constant.


Follicular traction urticaria.
Inducible urticaria is a heterogeneous subgroup of chronic urticarias caused by a wide variety of environmental stimuli, such as exercise, cold, heat, pressure, sunlight, vibration, and water. A new term, follicular traction urticaria, was suggested as an unusual form of inducible urticarias. We report a patient who was diagnosed with follicular traction urticaria.


Striatal D2 receptors in symptomatic and asymptomatic carriers of dopa-responsive dystonia measured with [11C]-raclopride and positron-emission tomography.
We tested the hypothesis that asymptomatic carriers of dopa-responsive dystonia (DRD) have increased dopamine D2 receptors in the striatum that protect them from the clinical manifestations of dopaminergic deficiency. We examined striatal D2-receptor binding in (1) symptomatic subjects (treated and untreated) and (2) asymptomatic gene carriers. Using [11C]-raclopride PET, we found elevated striatal D2-receptor binding in both groups. In one of our drug-naive symptomatic subjects, 7 months of treatment with levodopa/carbidopa did not affect the receptor binding as measured on a second scan. We conclude that increased D2-receptor binding in DRD may be a homeostatic response to the dopaminergic deficit in subjects carrying the DRD gene, but is not the sole factor determining the clinical state of these individuals.


Alpha interferon administration to infants with congenital rubella.
Three infants with congenital rubella syndrome were given human leukocyte (alpha) interferon at doses of 2 x 10(5) to 7 X 10(5) U/kg per day for 10 days. A transient decrease in pharyngeal virus excretion was observed with treatment. No significant side effects were associated with the administration of human leukocyte interferon to these infants.


Longitudinal decline in speech production in Parkinson's disease spectrum disorders.
We examined narrative speech production longitudinally in non-demented (n=15) and mildly demented (n=8) patients with Parkinson's disease spectrum disorder (PDSD), and we related increasing impairment to structural brain changes in specific language and motor regions. Patients provided semi-structured speech samples, describing a standardized picture at two time points (mean±SD interval=38±24months). The recorded speech samples were analyzed for fluency, grammar, and informativeness. PDSD patients with dementia exhibited significant decline in their speech, unrelated to changes in overall cognitive or motor functioning. Regression analysis in a subset of patients with MRI scans (n=11) revealed that impaired language performance at Time 2 was associated with reduced gray matter (GM) volume at Time 1 in regions of interest important for language functioning but not with reduced GM volume in motor brain areas. These results dissociate language and motor systems and highlight the importance of non-motor brain regions for declining language in PDSD.


Phi-X-174 bacteriophage structural mutants which affect deoxyribonucleic acid synthesis.
Seven cistrons in phiX-174 were identified and one in particular was studied intensively: cistron A, which is assigned a protein in the mature phage. Amber mutants in this cistron synthesize a new deoxyribonucleic acid (DNA) form in addition to circular phage DNA upon infection of the restrictive host. This DNA is linear, non-infectious, and single-stranded; it is formed from the phage strand of replicative form phiX-174 DNA. These mutants produce two different defective particles in the restrictive host. One particle contains circular phage DNA but is not infectious; the other contains the new DNA form and is similar to the 70S particles found in wild-type phage lysates. The mutant A gene product acts independently of normal A protein upon mixed infection of the restrictive host with an A mutant and a mutant from any other cistron or wild type.


Isoiridomyrmecin rich essential oil from Nepeta erecta Benth. and its antioxidant activity.
The essential oil composition of the aerial parts of Nepeta erecta Benth. (Family: Lamiaceae) from Uttarakhand, India was analysed by capillary gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). A total of 34 constituents were identified representing 94.6% of the oil composition. Oxygenated monoterpenes (74.0%) constituted the major proportion of the oil, dominated by isoiridomyrmecin (70.6%) as a single major constituent. Other significant constituents were caryophyllene oxide (9.6%), β-Bourbonene (2.0%), humulene epoxide II (1.7%) and linalool (1.0%). The in vitro antioxidant activity (AOA) was assessed using β-Carotene bleaching assay, reducing power, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and inhibition of lipid peroxidation methods. The oil was found to exhibit AOA by inhibiting β-Carotene bleaching (54.6 ± 2.52%) and by scavenging DPPH free radical (IC(50) = 0.74 ± 0.12 mg mL(-1)). The AOA of the essential oil of N. erecta and its major compound isoiridomyrmecin has not been reported so far.


Phosphoenolpyruvate carboxylase of Escherichia coli. Inhibition by various analogs and homologs of phosphoenolpyruvate.
In an attempt to investigate the topography of the catalytic site of phosphoenolpyruvate (PEP) carboxylase [EC 220.127.116.11] of Escherichia coli, the inhibitor constants (Ki) for more than 20 compounds were determined with the reaction system containing dioxane, a non-physiological activator of the enzyme. The Ki values for the compounds lacking methylene-, carboxylate-, or phosphate groups were all more than 10-fold larger than the Km value for PEP, indicating the significant contribution of these groups to the binding of PEP with the enzyme. The Ki value for L-phospholactate (0.30 mM) was almost equal to the Km value for PEP (0.25 mM), whereas that for D-phospholactate (0.89 mM) was about 3-fold larger than the Km value. It was presumed that PEP binds with the enzyme on its si-side. Among 6 PEP homologs, the Ki values for phosphoenol alpha-ketobutyrate (0.024 mM) and phosphoenol alpha-ketovalerate (0.034 mM) were about one-tenth the Km value, indicating the presence of a hydrophobic pocket around the binding site of the methylene group of PEP, where the carboxylation reaction is supposed to occur. DL-Phosphomalate, a presumptive carboxylated substrate, was a weak inhibitor with a Ki value of 2.20 mM.


The Drosophila RASSF homolog antagonizes the hippo pathway.
Correct organ size is determined by the balance between cell death and proliferation. Perturbation of this delicate balance leads to cancer formation . Hippo (Hpo), the Drosophila ortholog of MST1 and MST2 (Mammalian Sterile 20-like 1 and 2) is a key regulator of a signaling pathway that controls both cell death and proliferation . This pathway is so far composed of two Band 4.1 proteins, Expanded (Ex) and Merlin (Mer), two serine/threonine kinases, Hpo and Warts (Wts), the scaffold proteins Salvador (Sav) and Mats, and the transcriptional coactivator Yorkie (Yki). It has been proposed that Ex and Mer act upstream of Hpo, which in turn phosphorylates and activates Wts. Wts phosphorylates Yki and thus inhibits its activity and reduces expression of Yki target genes such as the caspase inhibitor DIAP1 and the micro RNA bantam. However, the mechanisms leading to Hpo activation are still poorly understood. In mammalian cells, members of the Ras association family (RASSF) of tumor suppressors have been shown to bind to MST1 and modulate its activity . In this study, we show that the Drosophila RASSF ortholog (dRASSF) restricts Hpo activity by competing with Sav for binding to Hpo. In addition, we observe that dRASSF also possesses a tumor-suppressor function.


[Gastrointestinal hemorrhage after operation for dissecting aneurysm of the thoracic aorta].
We present the case of a 45-year-old man who underwent surgical repair of a dissecting aneurysm of the descending thoracic aorta. 25 days postoperatively, the patient unexpectedly developed hematemesis and hypovolemic shock. Emergency gastroscopy was performed and a suspected aortoesophageal fistula was diagnosed. Unfortunately, the patient died prior to emergency surgery. Aneurysms of the aorta or of aortic grafts occasionally lead to the development of fistulae to the upper gastrointestinal tract. Once this rare, but life-threatening complication is suspected, endoscopy must be performed immediately. Emergency surgery is associated with a high risk, but represents the only possibly chance of survival for the patient.


Hydrothermal synthesis, characterization and photocatalytic properties of Cu2PO4OH with hierarchical morphologies.
Cu2PO4OH with various hierarchical morphologies were synthesized using a hydrothermal method by simple adjusting the pH value and temperature. The prepared powders were characterized by X-ray diffraction (XRD), Raman spectroscopy, fourier-transform infrared spectroscopy (FT-IR), and UV-Vis diffuse reflectance spectroscopy. It was found that overall morphologies of Cu2PO4OH were varied from micro-rods to walnut-like microspheres as increasing the initial solution pH value and their size of the assembled crystallites was smaller at lower temperature. From the FT-IR and Raman analysis, the incorporation of the hydroxyl ions (OH-) into the crystal structure plays important roles to their crystallization process, and finally affects their morphologies. Moreover, the optical absorption properties of the Cu2PO4OH were strongly related to their morphologies. The photocatalytic activity of Cu2PO4OH with various morphologies was also investigated via the degradation of rhodamine B (RhB) dye solution under visible light irradiation (lambda > 410 nm). The walnut-like microsphere assembled with nanoparticles exhibited higher photocatalytic activity due to the higher optical absorption as well as a higher surface area.


Splenectomy for splenic metastases from malignant adrenal pheochromocytoma: a case report.
Splenic metastasis is generally not a common clinical event. However, metastasis to the spleen from adrenal pheochromocytoma is extremely rare and has not been reported in literature. This report presents a case of a 58 year-old male patient who developed spleen-only metastases in July 2007. The patient had a previous history of left epinephroectomy for adrenal pheochromocytoma in January 2003. Abdominal computed tomography demonstrated multiple enhancing lesions suggestive of metastases; thus splenectomy was performed. Pathological examinations confirmed the diagnosis of splenic metastases from pheochromocytoma. The patient was alive without recurrence 48 months after splenectomy. This study is the first report on splenic metastasis from previous adrenal pheochromocytoma, and long-term survival was achieved by splenectomy. A history of malignancy indicates a high index of suspicion for splenic metastasis, and long-term survival can be achieved by splenectomy for spleen-only metastasis.


Oxygen Vacancy Engineering of Fe-Doped NiMoO4 for Electrocatalytic N2 Fixation to NH3.
Electrochemical nitrogen reduction reaction (NRR) is a promising method for ammonia synthesis under ambient conditions. However, the NRR performance is limited to an extremely strong N≡N bond in N2 and the competing hydrogen evolution reaction. Introducing oxygen vacancies (OVs) has been considered as a forceful means to accelerate the sluggish NRR reaction kinetics. Herein, we reported the design of Fe-doped NiMoO4 catalysts for NRR. Fe doping can increase the amount of OVs in the catalyst and contribute to lattice strain enhancement, thereby leading to the improvement of the electron transport rate and catalytic active for NRR. In 0.1 M Na2SO4 solution, the 5% Fe-NiMoO4 catalyst achieves a NH3 yield rate of 15.36 μg h-1 mgcat.-1 and a Faradaic efficiency of 26.85% under -0.5 V versus RHE. Furthermore, the 5% Fe-NiMoO4 catalyst exhibits excellent stability (up to 13 h) during the reaction.


Premolar root morphology and metric variation in Pan troglodytes verus.
Premolar root form remains an important taxonomic character in hominin alpha taxonomy. Variation in detailed aspects of root structure remains poorly characterized in extant apes. This limited comparative context hinders evaluations of the significance of root form variation in hominin systematics. Using micro-computed tomography we examine morphological variation in premolars in 51 (n = 128 premolars) West African chimpanzees (Pan troglodytes verus). We categorize premolar root/canal form and number, based on the external root surface and pulp canal morphology, and test for a relationship between canal configuration and sex, jaw size, tooth/root size and cervix shape. Jaw size and root size/shape were quantified using standard metrics, and geometric morphometrics was used to examine root form and cervix shape. Our results confirm previous findings in external root form, but reveal previously undocumented variation in mandibular premolar canal number/form in this subspecies. The LP3 and UP4 exhibit variation in canal number/form, while the UP3 is restricted to external root configurations. The LP4 expresses only a single root/canal form. Generally, in LP3 and UP4 there is no correlation between canal form/number and sex, root size, and jaw size; UP4 canal variation covaried with cervix shape and size. Cervix size is significantly greater in three canal UP4s than two canal UP4s. Our results highlight canal form/number as an important aspect when characterizing root form. The implications of our results for the taxonomic utility of root form within the hominoid and hominin clades are discussed.


Long-term follow-up in managing anaplastic astrocytoma by multimodality approach with surgery followed by postoperative radiotherapy and PCV-chemotherapy: phase II trial.
Overall survival and progression-free survival after 5 and 10 years of 31 patients with malignant glioma treated by a combination of surgery, postoperative radiotherapy, and chemotherapy with a PCV regimen (procarbazine, CCNU [lomustine] and vincristine) is described. Parameters were evaluated by age at diagnosis, gender, ethnic origin, pre- and postsurgery Karnofsky Performance Status (KPS) score, limit and amount of surgical resection, histopathologic type, number of chemotherapy courses, time between surgery and radiotherapy, response to combined therapy, and dosage and type of radiotherapy. Progression-free survival was 29% at 24 months and 22% at 60 and 120 months. Overall survival was 47%, 36%, and 36% after 24, 60, and 120 months, respectively. Favorable prognostic factors for survival in univariate analysis were pre- and postoperative KPS (> or =70; p = 0.015; p = 0.0025, respectively), age of patients (<40; p = 0.01), number of chemotherapy cycles (> or =6; p = 0.02), and radiation dose (> or =60 Gy; p = 0.0015). The only significant prognostic factors for overall survival in a stepwise multivariate analysis were irradiation dose (p = 0.0001), number of chemotherapy cycles (p = 0.001), and preoperative KPS (p = 0.05); for progression-free survival it was number of chemotherapy cycles (p = 0.004). Survival was not affected by excision size, radiation method, histopathologic type of tumor, gender, ethnic origin, or time lapsed between surgery and irradiation.


LBH589 enhances T cell activation in vivo and accelerates graft-versus-host disease in mice.
Histone deacetylase inhibitors (HDACis) are a new class of compounds that induce acetylation of histone lysine tails in chromatin and modify gene expression. The Food & Drug Administration approved HDACi, Vorinostat, or suberoylanilide hydroxamic acid (SAHA), has been shown to inhibit tumor cell growth and the production of proinflammatory cytokines. In preclinical allogeneic transplant models, SAHA induces graft-versus-host disease (GVHD) amelioration in treated mice without impairing graft-versus-leukemia. LBH589 (Panobinostat), a structurally novel cinnamic hydroxamic acid class, is an HDACi more potent than SAHA. In the current work, we tested the hypothesis that LBH589 would be highly effective in the prevention of GVHD. Using mouse model of allogeneic bone marrow transplant (BMT), we unexpectedly found that treatment with LBH589 accelerated GVHD, in contrast to the treatment with SAHA that alleviated GVHD. Accelerated GVHD in the recipients treated with LBH589 was associated with elevated Th1 cytokines in recipient serum, enhanced CXCR3 expression on donor T cells, and T cell infiltration in the liver. The current study highlights the distinct effects of pan HDACi on allogeneic BMT and alerts that LBH589 (Panobinostat) could have an adverse effect on GVHD, and possibly on other inflammatory diseases.


[Mismatch of ASPECTS based on arterial spin labeling and diffusion-weighted imaging as an indicator for mechanical thrombectomy in patients with wake-up stroke].
To retrospectively analyze the outcomes of wake-up stroke (WUS) patients with occlusion of large vessel occlusion (LVO), who were selected for mechanical thrombectomy according to the mismatch of Alberta Stroke Program Early CT Score (ASPECTS) based on arterial spin labeling (ASL) and diffusion-weighted image (DWI) on admission magnetic resonance (MR) scans. Twelve consecutive WUS patients with acute LVO of the anterior circulation undergoing MR scans with ASL and DWI prior to thrombectomy were retrospectively evaluated. The mismatch of ASPECTS was defined as the difference between ASL-ASPECTS and DWI-ASPECTS, and a higher score indicates a greater mismatch. The procedures led to successful reperfusion in all the cases (Thrombolysis in Cerebral Infarction Grade 2b-3). Eleven patients (91.7%) had significantly decreased National Institute of Health Stroke scale (NIHSS) score at discharge.AmRS score of ≤2 at 90 days was achieved in 8 of the 12 patients (66.7%). The mismatch between ASPECTS assessed based on ASL and DWI can detect a true mismatch in patients with acute LVO of the anterior circulation, and can be used for rapid screening of patients eligible for thrombectomy.


Effect of moderate prenatal ethanol exposure on the differential expression of two histamine H3 receptor isoforms in different brain regions of adult rat offspring.
We have reported that prenatal alcohol exposure (PAE) elevates histamine H3 receptor (H3R) agonist-mediated inhibition of glutamatergic neurotransmission in the dentate gyrus. Here, we hypothesized that PAE alters the expression of two prominent H3R isoforms namely, the rH3A and rH3C isoforms, which have differing intrinsic activities for H3R agonists, in a manner that may contribute to heightened H3R function in PAE rats. In contrast to our predictions, we found different effects of sex and PAE in various brain regions with significant interactions between sex and PAE in dentate gyrus and entorhinal cortex for both isoforms. Subsequently, to confirm the PAE-and sex-induced differences on H3R isoform mRNA expression, we developed a polyclonal antibody selective for the rH3A inform. Western blots of rH3A mRNA-transfected HEK-293 cells identified a ~ 48 kDa band of binding consistent with the molecular weight of rH3A, thus confirming antibody sensitivity for rH3A protein. In parallel, we also established a pan-H3R knockout mice line to confirm antibody specificity in rodent brain membranes. Both qRT-PCR and H3R agonist-stimulated [35S]-GTPγS binding confirmed the absence of mH3A mRNA and H3 receptor-effector coupling in H3R knockout (KO) mice. Subsequent western blotting studies in both rat and mouse brain membranes were unable to detect rH3A antibody binding at ~48 kDa. Rather, the H3RA antibody bound to a ~ 55 kDa band in both rat and mouse membranes, including H3R KO mice, suggesting H3RA binding was not specific for H3Rs in rodent membranes. Subsequent LC/MS analysis of the ~55 kDa band in frontal cortical membranes identified the highly abundant beta subunit of ATPase in both WT and KO mice. Finally, LC/MS analysis of the ~48 kDa band from rH3A mRNA-transfected HEK-293 cell membranes was able to detect rH3A protein, but its presence was below the limits of quantitative reliability. We conclude that PAE alters rH3A and rH3C mRNA expression in some of the same brain regions where we have previously reported PAE-induced alterations in H3R-effector coupling. However, interpreting the functional consequences of altered H3R isoform expression was limited given the technical challenges of measuring the relatively low abundance of rH3A protein in native membrane preparations.


Mechanistic insights into sequestration of U(VI) toward magnetic biochar: Batch, XPS and EXAFS techniques.
The magnetic iron oxide (Fe3O4) nanoparticles stabilized on the biochar were synthesized by fast pyrolysis of Fe(II)-loaded hydrophyte biomass under N2 conditions. The batch experiments showed that magnetic biochar presented a large removal capacity (54.35mg/g) at pH3.0 and 293K. The reductive co-precipitation of U(VI) to U(IV) by magnetic biochar was demonstrated according to X-ray diffraction, X-ray photoelectron spectroscopy and X-ray absorption near edge structure analysis. According to extended X-ray absorption fine structure analysis, the occurrence of U-Fe and U-U shells indicated that high effective removal of uranium was primarily inner-sphere coordination and then reductive co-precipitation at low pH. These observations provided the further understanding of uranium removal by magnetic materials in environmental remediation.


LigB subunit vaccine confers sterile immunity against challenge in the hamster model of leptospirosis.
Neglected tropical diseases, including zoonoses such as leptospirosis, have a major impact on rural and poor urban communities, particularly in developing countries. This has led to major investment in antipoverty vaccines that focus on diseases that influence public health and thereby productivity. While the true, global, impact of leptospirosis is unknown due to the lack of adequate laboratory diagnosis, the WHO estimates that incidence has doubled over the last 15 years to over 1 million cases that require hospitalization every year. Leptospirosis is caused by pathogenic Leptospira spp. and is spread through direct contact with infected animals, their urine or contaminated water and soil. Inactivated leptospirosis vaccines, or bacterins, are approved in only a handful of countries due to the lack of heterologous protection (there are > 250 pathogenic Leptospira serovars) and the serious side-effects associated with vaccination. Currently, research has focused on recombinant vaccines, a possible solution to these problems. However, due to a lack of standardised animal models, rigorous statistical analysis and poor reproducibility, this approach has met with limited success. We evaluated a subunit vaccine preparation, based on a conserved region of the leptospiral immunoglobulin-like B protein (LigB(131-645)) and aluminium hydroxide (AH), in the hamster model of leptospirosis. The vaccine conferred significant protection (80.0-100%, P < 0.05) against mortality in vaccinated animals in seven independent experiments. The efficacy of the LigB(131-645)/AH vaccine ranged from 87.5-100% and we observed sterile immunity (87.5-100%) among the vaccinated survivors. Significant levels of IgM and IgG were induced among vaccinated animals, although they did not correlate with immunity. A mixed IgG1/IgG2 subclass profile was associated with the subunit vaccine, compared to the predominant IgG2 profile seen in bacterin vaccinated hamsters. These findings suggest that LigB(131-645) is a vaccine candidate against leptospirosis with potential ramifications to public and veterinary health.


A case of primary pericardial undifferentiated sarcoma.
Primary pericardial sarcomas are extremely rare. The authors report a case of a 46-yr-old woman in whom a large mediastinal mass was discovered. The patient presented with cough, dyspnea, and orthopnea. Diagnostic investigations, such as echocardiography, computed tomography, and exploratory thoracotomy provided the evidence of a large mass in the mediastinum, attached by a broad base to the superior portion of the pericardium. A excisional biopsy was performed, and histologic examination of a biopsy specimen showed undifferentiated sarcoma. However, the complete removal of the mass was impossible due to adhesion to the adjacent great vessels. After the completion of the chemotherapy the patient was completely asymptomatic. However, follow-up transesophageal echocardiography showed a residual 3 x 4 cm-sized mass. The patient received the radiotherapy with a total dose of 55 Gy over 6 weeks. At present, there is no evidence of disease progression.


The cerebro-costo-mandibular syndrome: seven patients and review of the literature.
We report seven patients with the cerebro-costo-mandibular syndrome, a multiple congenital anomaly syndrome with, as the most distinctive features extreme micrognathia and abnormal rib development. Twice a parent to child transmission was found, compatible with autosomal dominant mode of inheritance. One of our patients presented with absence of the auditory canals and subluxation of the radial head, and another with choanal atresia, findings that have not been published previously. Longterm follow-up of some of the patients is described, and an overview of the literature is given.


Emergence of photoswitchable states in a graphene-azobenzene-Au platform.
The perfect transmission of charge carriers through potential barriers in graphene (Klein tunneling) is a direct consequence of the Dirac equation that governs the low-energy carrier dynamics. As a result, localized states do not exist in unpatterned graphene, but quasibound states can occur for potentials with closed integrable dynamics. Here, we report the observation of resonance states in photoswitchable self-assembled molecular(SAM)-graphene hybrid. Conductive AFM measurements performed at room temperature reveal strong current resonances, the strength of which can be reversibly gated on- and off- by optically switching the molecular conformation of the mSAM. Comparisons of the voltage separation between current resonances (∼ 70-120 mV) with solutions of the Dirac equation indicate that the radius of the gating potential is ∼ 7 ± 2 nm with a strength ≥ 0.5 eV. Our results and methods might provide a route toward optically programmable carrier dynamics and transport in graphene nanomaterials.


Multidrug-resistant tuberculosis outbreak among transvestite sex workers, Buenos Aires, Argentina.
We describe the first outbreak of multidrug-resistant tuberculosis (MDR-TB) that occurred in Argentina among transvestite sex workers, and actions undertaken for its control. In Buenos Aires city, transmission was documented between 2001 and 2004 by conventional and molecular methods in a hotel where transvestites used to reside and work. The source case was traced back to 1998. Six secondary cases were diagnosed and treated. Thirty-two contacts were investigated. The outbreak strain had formerly caused nosocomial transmission in Rosario, a city 300 km from Buenos Aires. Our findings highlight the difficulties controlling MDR-TB in Argentina.


Effects of alliin on LPS-induced acute lung injury by activating PPARγ.
Alliin is a garlic organosulfur compound that possesses various pharmacological properties. In the present study, the protective effects and molecular mechanism of alliin on Lipopolysaccharides (LPS)-induced acute lung injury (ALI) were analyzed. LPS-induced ALI was induced in BALB/c mice by intranasal instillation of LPS. Alliin was administered intraperitoneally to mice 1 h after LPS treatment. The results showed that alliin markedly inhibited lung myeloperoxidase (MPO) activity and wet/dry (W/D) ratio induced by LPS. Alliin also inhibited TNF-α and IL-1β in the bronchoalveolar lavage fluid (BALF) induced by LPS. Furthermore, LPS-induced lung pathological injury was attenuated by treatment of alliin. LPS-induced NF-κB activation was significantly inhibited by alliin. In addition, the expression of peroxisome proliferator-activated receptor γ (PPARγ) was up-regulated by treatment of alliin. Taken together, these results suggested that alliin protected against LPS-induced ALI by activating PPARγ, which subsequently inhibited LPS-induced NF-κB activation and inflammatory response. Alliin might be used as an anti-inflammatory agent in the treatment of ALI.


A novel neutralizing monoclonal antibody against cell-binding polypeptide of ricin.
One strain of neutralizing monoclonal antibody (MAb) against cell-binding polypeptide of ricin, named 3E1, was generated efficiently. The antibody recognized the linearity epitope of RTB located in a toxin structure domain characterized by Western blotting. The safe period of mice for intraperitoneal injection of 100 microg of antibody was 20 min after intraperitoneal injection of 2 microg of Ricin (10 times LD50). The neutralizing MAb we obtained could be developed into an immunotherapeutic agent to counteract the use of ricin as a terrorist or biological warfare weapon. It might be useful, as well, for antibody-based prophylaxis.


Follow-up Care among Medicaid Patients with Schizophrenia treated with Antipsychotics in the Inpatient Setting.
Background: For patients with schizophrenia, the transition from inpatient hospitalization to outpatient care presents a challenge to providing continuous care. Lapses in care during this time period can result in poor clinical outcomes. To date, there is little information regarding the association between inpatient antipsychotic treatment and outpatient care. Objectives: The objectives of this study were to describe trends in and identify factors associated with post-discharge follow-up outpatient care among Medicaid enrollees with schizophrenia treated with antipsychotics. Methods: Adults administered oral or long-acting injectable antipsychotic medication during a hospitalization for schizophrenia were identified in the linked MarketScan® Hospital Drug and Multi-State Medicaid Databases. Psychiatric-related follow-up outpatient visits within 30 days of discharge were identified from Medicaid claims based on Healthcare Effectiveness Data and Information Set specifications. Kaplan-Meier curves and Cox proportional hazards models were used to describe and analyze time to follow-up visit and to identify patient and hospitalization characteristics associated with follow-up visit. Results: The study sample (N=1,312) had a mean age of 40.5 years and was 57% male. A follow-up outpatient visit was identified among 47% of patients. The proportion of patients with a follow-up visit ranged from 25% in 2005 to 48% in 2010/2011. The Cox proportional hazard model suggests that capitated health plan, attending physician specialty of psychiatry/psychology, and later year of index hospitalization significantly increase the probability of a follow-up visit, while substance-related disorders significantly decrease the probability. Type of antipsychotic received during index hospitalization was not significantly associated with probability of a follow-up visit. Conclusion: While follow-up visit rates have increased over time, this study highlights the ongoing need for improvements in effective linkage to outpatient care for patients hospitalized and treated for schizophrenia, particularly among patients with comorbid substance abuse disorder.


Biological Activities of Libidibia (Caesalpinia) ferrea var. parvifolia (Mart. ex Tul.) L. P. Queiroz Pod Preparations.
Libidibia ferrea has been used in folk medicine throughout Brazil, and this study evaluated the biological activities of crude extract (CE) as well as a partially purified fraction (F80) obtained from its pods. Results from the MTT assay revealed that only F80 inhibited NCI-H292 cell growth; however, neither CE nor F80 reduced HEp-2 cell growth or sarcoma 180 tumor weight with the in vivo assay. Acute oral toxicity of the extract and fraction was evaluated following the steps of Guideline 423, using female mice; LD(50) for both preparations was determined as 2,500 mg/kg body weight. CE and F80 promoted a reduction of the leukocyte number and nitrite level in inflammatory exudates when the anti-inflammatory assay (carrageenan-induced peritonitis) was performed. CE and F80 inhibited writhing regarding antinociceptive activity (acetic acid-induced writhing response in mice). In conclusion, CE and F80 have no significant cytotoxic or antitumor activities in cell lines showing low toxicity and no action against tumors in vivo. Both preparations revealed anti-inflammatory and antinociceptive activities, corroborating the pharmacological basis of L. ferrea for ethnomedical use.


Antiphotoaging effect of conditioned medium of dedifferentiated adipocytes on skin in vivo and in vitro: a mechanistic study.
Photoaging of skin occurs partially due to decreased synthesis and increased degradation of dermal collagen. Antiphotoaging therapy aims to counteract these effects. This study aimed to investigate whether secretory factors from dedifferentiated adipocytes (DAs) could alleviate photoaging in human dermal fibroblasts (HDFs) and in mice and to clarify the underlying mechanism. DAs were acquired and verified based on cellular biomarkers and multilineage differentiation potential. The concentrations of several cytokines in conditioned medium from DAs (DA-CM) were determined. In vivo pathological changes, collagen types I and III, and matrix metalloproteinase (MMP)-1 and -3 were evaluated following the injection of 10-fold concentrated DA-CM into photoaged mice. In vitro, the effect of DA-CM on stress-induced premature senescence in HDFs was investigated by 5-ethynyl-2'-deoxyuridine (EdU) staining and β-galactosidase staining. The influence of DA-CM and transforming growth factor-β1 (TGF-β1) on the secretion of collagen types I and III, MMP-1, and MMP-3 in HDFs was evaluated by ELISA. In vivo, we found that subcutaneously injected 10-fold concentrated DA-CM increased the expression of collagen types I and III. In vitro, DA-CM clearly mitigated the decreased cell proliferation and delayed the senescence status in HDFs induced by ultraviolet B (UVB). HDFs treated with DA-CM exhibited higher collagen types I and III secretion and significantly lower MMP-1 and MMP-3 secretion. The TGF-β1-neutralizing antibody could partially reduce the recovery effect. Our results suggest that DAs may be useful for aging skin and their effects are mainly due to secreted factors, especially TGF-β1, which stimulate collagen synthesis and alleviate collagen degradation in HDFs.


Identification of differentially expressed inflammatory factors in Wilms tumors and their association with patient outcomes.
The present study aimed to identify differentially expressed inflammatory factors observed in Wilms tumors (WT), and to investigate the association of these factors with clinical stage, pathological type, lymph node metastasis and vascular involvement of WT. Surface-enhanced laser desorption/ionization-time of flight mass spectrometry was performed to screen differentially expressed proteins among WT and normal tissue pairs. Upregulated proteins in WT were separated and purified by solid phase extraction and Tricine SDS-PAGE, respectively. Following in-gel digestion, the peptide mixture was subjected to liquid chromatography mass spectrometry to identify proteins on the basis of their amino acid sequences. Immunohistochemistry was used to confirm the expression of differentially expressed inflammatory proteins. Of the proteins that were upregulated in WT, two proteins with mass/charge (m/z) ratio of 12,138 and 13,462 were identified as macrophage migration inhibitory factor (MIF) and C-X-C motif ligand 7 (CXCL7) chemokine, respectively. The expression of these two proteins was increased in WT compared with adjacent normal tissues and normal renal tissues, and increased with increasing clinical stage. In addition, their expression was significantly increased in patients with unfavorable pathological type, lymph node metastasis and vascular involvement compared with the groups with favorable type, and without lymph node metastasis or vascular involvement (P<0.05). Increased pro-inflammatory MIF and CXCL7 expression in WT is closely associated with the clinical stage, pathological type, lymph node metastasis and vascular involvement, and may represent biomarkers for the clinical diagnosis of WT.


Disentangling the impact of resistance and ambivalence on therapy outcomes in cognitive behavioural therapy for generalized anxiety disorder.
Resistance and ambivalence about change are increasingly recognized as important determinants of treatment outcomes. Moreover, resistance and ambivalence are thought to be theoretically related in that clients who are more ambivalent about change are more likely to demonstrate resistance to the process and tasks of treatment. In the context of cognitive behavioural therapy (CBT) for generalized anxiety disorder, the present study simultaneously examined early resistance and ambivalence using two observer-based coding systems in order to determine their inter-relationship and, importantly, to investigate their relative contributions to outcome. Resistance was also coded during mid-treatment in order to investigate possible mediation pathways. Early ambivalence (clients' arguments against change or counter-change talk) was found to be no longer related to outcomes when early resistance was taken into account, suggesting that disharmony in the therapeutic relationship is more important to outcomes than ambivalence per se. Moreover, mid-treatment resistance partially mediated the relationship between early resistance and post-treatment worry severity. That is, higher early opposition to therapist direction is related to poorer outcomes, in part because it is associated with greater resistance during the working phase of CBT. The findings underscore the critical need for therapists to be sensitive to identifying resistance early and throughout treatment.


[Moroccan experience in the surgical treatment of multiple hydatid cysts in the liver].
We studied 94 cases of multiple hydatid cysts in the liver, over a period of ten years. These cases accounted for 31.3% of all cases of hydatid cysts treated surgically in the Visceral Surgery Department of Avicenne Military Hospital in Marrakech. In these patients, who were often young and male, the principal symptoms were pain in the right hypochondrium (71.3%) and hepatomegaly (24.5%). In about 10% of cases, the cysts were discovered by chance. Ultrasound and CT scans facilitated diagnosis and determination of the position of the cysts, with reliability reaching 100% for CT scans. The cysts had burst in the bile ducts in 26.6% of cases and were infected in 8 cases. They were multivesicular in 77.5% of cases. Association with hydatidosis at another site was observed in 28 cases: in the peritoneum in 15, the thorax in 7, the diaphragm in 4, the spleen in 2 and the kidney in 1 case. Surgically, the route most frequently used was double incision below the rib cage (49.5%). It is not possible to recommend one particular way to treat cysts and the most appropriate approach to treatment depends on the site, type and number of cysts. Resection of the prominent dome is the technique most frequently used (57.25%). However, in recent years, the use of cystectomy has been increasing (20.2%) due to the considerable decreases in post-operative morbidity and duration of hospital stay that it affords. The principal post-operative complications observed were abscesses under the diaphragm (6 cases), biliary leakage (5 cases), pleurisy (6 cases) and the formation of abscesses in the vestigial cavity (4 cases). The rate of morbidity in the RDS appeared high, accounting for 75% of total morbidity. Only one patient died. This patient died from severe hepatic insufficiency due to the near destruction of the liver by the hydatosis. We observed two recurrences during follow up. Both underwent further surgery and neither suffered complications.


Spinal manipulation impacts cervical spine movement and fitts' task performance: a single-blind randomized before-after trial.
The objective of this study was to determine if active cervical range of motion (ROM) and Fitts' task movement time differences occurred after high-velocity low-amplitude cervical spinal manipulation (SM) across various indexes of difficulty. A single-blind randomized before-after trial was performed in a motor performance laboratory. Fifteen volunteers (21-42 years) with asymptomatic palpable intervertebral motion restriction at the C1-C2 level were randomly assigned to an SM group or to a no-intervention (NI) group. A single episode of upper cervical manipulation was performed on the SM group. Active cervical ROM and movement time were measured pre and posttreatment in the SM group and compared to similar measurements in the NI group. In the SM group, active cervical ROM into rotation increased after the intervention (pre, 74.75 degrees +/- 7.63 degrees ; post, 78.50 degrees +/- 7.23 degrees ; t (7) = -3.07; P < .02). During the second trial, significant group differences were present in the SM group for movement time in direction congruent conditions (F((8,48)) = 2.83; P < .02; eta(p)(2) = .320) and direction incongruent conditions (F((8,48)) = 2.31; P < .05; eta(p)(2) = .278) but not for the NI group. A linear relationship between indexes of difficulty and movement time as predicted by Fitts' law was observed. Significant group effects indicate that SM not only increases cervical active ROM but also facilitates the performance of a cervical spine Fitts' task requiring rotation. This task may be used to quantify motor performance in clinically symptomatic populations with reduced ROM who are appropriate candidates for SM.


Effects of Cardiovascular Risk Factor Variability on Health Outcomes.
Innumerable studies have suggested "the lower, the better" for cardiovascular risk factors, such as body weight, lipid profile, blood pressure, and blood glucose, in terms of health outcomes. However, excessively low levels of these parameters cause health problems, as seen in cachexia, hypoglycemia, and hypotension. Body weight fluctuation is related to mortality, diabetes, obesity, cardiovascular disease, and cancer, although contradictory findings have been reported. High lipid variability is associated with increased mortality and elevated risks of cardiovascular disease, diabetes, end-stage renal disease, and dementia. High blood pressure variability is associated with increased mortality, myocardial infarction, hospitalization, and dementia, which may be caused by hypotension. Furthermore, high glucose variability, which can be measured by continuous glucose monitoring systems or self-monitoring of blood glucose levels, is associated with increased mortality, microvascular and macrovascular complications of diabetes, and hypoglycemic events, leading to hospitalization. Variability in metabolic parameters could be affected by medications, such as statins, antihypertensives, and hypoglycemic agents, and changes in lifestyle patterns. However, other mechanisms modify the relationships between biological variability and various health outcomes. In this study, we review recent evidence regarding the role of variability in metabolic parameters and discuss the clinical implications of these findings.


Mesenchymal stem cell-based bioengineered constructs: foreign body response, cross-talk with macrophages and impact of biomaterial design strategies for pelvic floor disorders.
An excessive foreign body response (FBR) has contributed to the adverse events associated with polypropylene mesh usage for augmenting pelvic organ prolapse surgery. Consequently, current biomaterial research considers the critical role of the FBR and now focuses on developing better biocompatible biomaterials rather than using inert implants to improve the clinical outcomes of their use. Tissue engineering approaches using mesenchymal stem cells (MSCs) have improved outcomes over traditional implants in other biological systems through their interaction with macrophages, the main cellular player in the FBR. The unique angiogenic, immunomodulatory and regenerative properties of MSCs have a direct impact on the FBR following biomaterial implantation. In this review, we focus on key aspects of the FBR to tissue-engineered MSC-based implants for supporting pelvic organs and beyond. We also discuss the immunomodulatory effects of the recently discovered endometrial MSCs on the macrophage response to new biomaterials designed for use in pelvic floor reconstructive surgery. We conclude with a focus on considerations in biomaterial design that take into account the FBR and will likely influence the development of the next generation of biomaterials for gynaecological applications.


Lack of evidence of transmission of HIV-1 to family contacts of HIV-1 infected children.
Although a number of studies have documented that casual household contact does not result in the transmission of HIV, isolated cases of person-to-person transmission have been reported. We report a study of household transmission in which the families were unaware the children were infected with HIV and thus took no precautions to prevent transmission. Twenty-two family members of nine transfusion-associated HIV-infected children were studied for transmission of HIV in households. There was a total of 174 person-year of household exposure; 76 of these exposure years were before the diagnosis of HIV infection in the index child. All family members tested negative for HIV by ELISA. Sharing household facilities, and interactions with the infected child including kissing, bathing, sleeping with, and helping to bathe, dress, and eat, did not result in transmission. Interactions that could theoretically result in person-to-person transmission occurred in these households such as caring for nose bleeds, biting, and home health care procedures. The findings of this and other studies support the participation of HIV-infected infants and children in out-of-home care programs. It remains prudent, however, to observe current recommendations for prevention of HIV-1 for all individuals regardless of whether HIV status is known.


Morphogenesis of rat lingual filiform papillae.
The morphogenesis of filiform papillae on rat tongue was investigated with the electron microscope. Tongue rudiments were first seen on the 12th day of gestation. At 15-17 days, dermal papillae had formed and were arranged in hexagonal array on the dorsal lingual surface. Capping each dermal papilla was a two-layered epithelium that protruded slightly above the lingual surface, thus forming the early filiform papilla. In the next stage of development, at 18-19 days of gestation, the epithelium lining the papilla had differentiated into two cell populations, one producing hard keratin, the other producing soft keratin. Some of the keratinized epithelial cells assumed a position at an acute angle to the tongue surface and extended deep into the epithelium. In the next stage, 20-21 days, a cleft appeared within these angularly oriented cells. This resulted in the division of the epithelium into keraatin-lined individual filiform papillae. Finally, the individual papillae increased in size to the adult form.


Human Mesenchymal Stem Cell-Derived Conditioned Media for Hair Regeneration Applications.
Hair loss can have major psychological impact on affected population belonging to varied ethnic background. Hair is a mini organ in itself and serves many distinguishing functions ranging from maintaining body temperature to promoting social interactions. Major cause of hair loss is androgenic alopecia. Hair follicles possess receptor for androgen. However, DHT (Dihydrotestosterone) in excess results into shrinkage of hair follicle affecting hair growth adversely. The present review is focused on etiology of hair loss, traditional treatment approach and their limitations with side effects with special emphasis on unique properties of stem cells, favourable growth factors secreted by stem cells and strategies to enhance favourable growth factor/cytokine production for hair loss therapeutics. We discussed in details the present available treatment options for hair loss like drugs (Finasteride and Minoxidil), follicular hair transplant, laser therapy and serum therapy. These treatment options have their own disadvantages and side effects with appropriate alerts from regulatory authorities. The side effects of these modalities cannot be ignored and demands alternate therapy approach with less or no side effects. We feel that the stem cell therapy is advancing and is a promising modality in near future owing to its advantages and promising outcomes. This review article discusses possible stem cell therapy for hair regrowth and its advantages. We focused on use of conditioned media derived from stem cells instead of using stem cells directly for the therapy.


A new factor controlling cell envelope integrity in Alphaproteobacteria in the context of cell cycle, stress response and infection.
The bacterial envelope is a remarkable and complex compartment of the prokaryotic cell in which many essential functions take place. The article by Herrou and collaborators (Herrou et al., in press), by a clever combination of structural analysis, genetics and functional characterization in free-living bacterial cells and during infection in animal models, elucidates a new factor, named EipA, that plays a major role in Brucella spp envelope biogenesis and cell division. The authors demonstrate a genetic connection between eipA and lipopolysaccharide synthesis, specifically genes involved in the synthesis of the O-antigen portion of lipopolysaccharide (LPS). Beyond its crucial role in Brucella physiology, the conservation of EipA in the class Alphaproteobacteria urges microbiologists to pursue future investigation of its homologs in other species belonging to this important group of bacteria.


A model-based simulator for testing rule-based decision support systems for mechanical ventilation of ARDS patients.
A model-based simulator was developed for testing rule-based decision support systems that manages ventilator therapy of patients with the Adult Respiratory Distress Syndrome (ARDS). The simulator is based on a multi-compartment model of the human body and mathematical models of the gas exchange abnormalities associated with ARDS. Initial testing of this system indicates that model-based simulators are a viable tool for testing rule-based expert systems used in health-care.