Simultaneous Quantification of Vesicle Size and Catecholamine Content by Resistive Pulses in Nanopores and Vesicle Impact Electrochemical Cytometry.
We have developed the means to simultaneously measure the physical size and count catecholamine molecules in individual nanometer transmitter vesicles. This is done by combining resistive pulse (RP) measurements in a nanopore pipet and vesicle impact electrochemical cytometry (VIEC) at an electrode as the vesicle exits the nanopore. Analysis of freshly isolated bovine adrenal vesicles shows that the size and internal catecholamine concentration of vesicles varies with the occurrence of a dense core inside the vesicles. These results might benefit the understanding about the vesicles maturation, especially involving the "sorting by retention" process and concentration increase of intravesicular catecholamine. The methodology is applicable to understanding soft nanoparticle collisions on electrodes, vesicles in exocytosis and phagocytosis, intracellular vesicle transport, and analysis of electroactive drugs in exosomes.


Condylomata of the uterine cervix and koilocytosis of cervical intraepithelial neoplasia.
In 202 women with koilocytotic atypia in cervical smears, 136 had predominantly small condylomata of the uterine cervix, and 66 had cervical intraepithelial neoplasia (CIN) of varying degree either with koilocytosis of the neoplasia or associated with condylomata. Koilocytosis correlated well with the histological diagnosis of condylomata, but occasionally it obscured the cytological evidence of CIN. Human papilloma virus particles were found in the cells of condylomata in 10 cases and in those of CIN II with koilocytosis in two cases of 21 examined ultrastructurally. There was evidence that the condyloma of the uterine cervix is a well-defined morphological entity and also that cytopathie changes similar to those seen in condylomata are present in some cases of CIN.


Bovine lymphosarcoma: expression of BLV-related proteins in cultured cells.
Bovine leukaemia virus (BLV) is known to be the aetiological agent of enzootic bovine lymphosarcoma. As the mechanism of tumour induction is unknown, we analysed the viral proteins expressed in cultured bovine cells of different origin, i.e. from enzootic or sporadic tumourous tissues, normal cells infected or not with BLV, and the reference FLK-BLV cells. We also investigated BLVs of different origins. As well as the previously described BLV polyproteins in FLK-BLV cells (pr70 and pr45) we have also found two additional polyproteins, p52 and p27. In these four proteins we observed the combined antigenicities of p24, p15 and p12. We observed an additional polyprotein in p42, with the antigenicities of the p24 and the p15 in cells derived from tumour tissue or infected in vitro with naturally occurring BLV isolates. None of these BLV-coded proteins, nor others with cross-reacting antigenicities, were encountered in non-BLV-producing cultured cells from enzootic or sporadic tumours. The use of autologous sera did not permit the detection of any proteins in addition to the above gag-coded and env-coded proteins. Thus, there is no evidence for the existence of a gag-x fusion protein which could play a role in BLV-induced tumourigenicity.


Massively parallel sample preparation for multiplexed single-cell proteomics using nPOP.
Single-cell proteomics by mass spectrometry (MS) allows quantifying proteins with high specificity and sensitivity. To increase its throughput, we developed nPOP, a method for parallel preparation of thousands of single cells in nanoliter volume droplets deposited on glass slides. Here, we describe its protocol with emphasis on its flexibility to prepare samples for different multiplexed MS methods. An implementation with plexDIA demonstrates accurate quantification of about 3,000 - 3,700 proteins per human cell. The protocol is implemented on the CellenONE instrument and uses readily available consumables, which should facilitate broad adoption. nPOP can be applied to all samples that can be processed to a single-cell suspension. It takes 1 or 2 days to prepare over 3,000 single cells. We provide metrics and software for quality control that can support the robust scaling of nPOP to higher plex reagents for achieving reliable high-throughput single-cell protein analysis.


A limited panel of immunomarkers can reliably distinguish between clear cell and high-grade serous carcinoma of the ovary.
The distinction of ovarian clear cell carcinomas (CCCs) from high-grade serous carcinomas (HG-SCs) is sometimes a diagnostic challenge. With the recognition that CCCs respond poorly to conventional chemotherapy there are efforts to initiate clinical trials for CCC, making accurate diagnosis critical. The purpose of this study was to test and validate a set of antibodies that could aid in the diagnosis of CCC, using a series of cases from different centers in North America. Using a test set of 133 CCCs, we identified the following markers: Cyclin E, estrogen receptor, hepatocyte nuclear factor (HNF)-1beta, Ki-67, p21, p53, and Wilms tumor (WT)1 that show significant discrimination from 200 HG-SCs. For validation, these markers were characterized on an independent set of 104 CCCs from 3 other centers. There were no significant differences in expression of these 7 markers between the independent test and validation sets of CCC. Combining all CCC cases (N=237), HNF-1beta showed the highest sensitivity (82.5%) and specificity (95.2%) for CCC, and WT1 for HG-SC (sensitivity: 79.9%, specificity: 97.4%). A diagnostic panel consisting of WT1, ER, and HNF-1beta demonstrated nearly identical performance as a panel using all 7 markers in distinguishing CCCs from HG-SCs, correctly classifying 84% of cases. Three percent of cases were misclassified and 13% carried an uninformative triple negative immunophenotype. CCCs show a distinct, reproducible immunophenotype, compared with HG-SCs, and a panel of 3 immunomarkers can serve as a diagnostic aid in problematic cases.


Ultrastructural changes in the liver after experimental spinal cord injury in rats: effects of methylprednisolone, immunoglobulin g and albumin.
Spinal Cord Injury (SCI) is routinely treated with standardized methyl prednisolone sodium succinate (MPSS) dose, so it is reassuring to find its effects on liver. We also evaluated the effects of albumin and immunoglobulin G (Ig G) therapies on liver if they are used in case of experimental SCI. The rats were allocated into six groups as control, trauma, vehicle, MPSS, Ig G and albumin consisting 8 rats for each. The rats with SCI were assigned to 30mg/kg MPSS, 5 mg/kg albumin and 400 mg/kg Ig G treatments. Tissue samples from liver were obtained for light and electron microscopy examinations and determination of myeloperoxidase (MPO) activity. Trauma increased MPO activity and caused cellular changes of liver tissue. Both albumin and Ig G treatments decreased MPO activity significantly. The light and electron microscopic evaluations showed remarkable preservation of liver ultra-structure with all treatments including MPSS. SCI resulted in neutrophil infiltration and changes in ultrastructure of liver. It was revealed that MPSS has no detrimental effects on liver. Although all treatments preserved liver tissue structure, Although all treatments preserved liver tissue structure, Ig G and albumin treatments also prevented neutrophil infiltration. To provide protection from secondary liver injury after SCI, use of albumin and Ig G treatments may be beneficial.


Hyperbolic spin vortices and textures in exciton-polariton condensates.
From cosmology to the microscopic scales of the quantum world, the study of topological excitations is essential for the understanding of phase conformation and phase transitions. Quantum fluids are convenient systems to investigate topological entities because well-established techniques are available for their preparation, control and measurement. Across a phase transition, a system dramatically changes its properties because of the spontaneous breaking of certain continuous symmetries, leading to generation of topological defects. In particular, attention is given to entities that involve both spin and phase topologies. Exciton-polariton condensates are quantum fluids combining coherence and spin properties that, thanks to their light-matter nature, bring the advantage of direct optical access to the condensate order parameter. Here we report on the spontaneous occurrence of hyperbolic spin vortices in polariton condensates, by directly imaging both their phase and spin structure, and observe the associated spatial polarization patterns, spin textures that arise in the condensate.


Detection of atypical porcine pestivirus genome in newborn piglets affected by congenital tremor and high preweaning mortality1.
Recently, piglets from a high-health status farm began exhibiting congenital tremors, high preweaning mortality and incidence of splayed legs. Postmortem histological examination identified a small number of scattered white matter vacuoles in the cerebellum and underlying brainstem of affected piglets. Presence of potential viral sources associated with this neurologic condition was initially infirmed using quantitative PCR for atypical porcine pestivirus (APPV), porcine teschovirus, and porcine sapelovirus. Using metagenomic analysis, APPV was identified as the main microbial species in serum obtained from piglets affected by congenital tremor. These piglets had higher preweaning mortality rates (46.4% vs. 15.3%) and incidence of splayed legs (33.0% vs. 0.8 %) compared to unaffected piglets. Piglets affected by congenital tremor had higher viral titer (P < 0.15) and larger birth weights (P < 0.05) compared to normal litter mates. Whole-genome sequencing and genome assembly of the novel APPV strain (MK728876) was carried out using Oxford Nanopore and related bioinformatics pipelines. Phylogenic analysis demonstrated that this strain along with other completely sequenced APPV strains were grouped into 2 clades, both including strains-inducing congenital tremor. Strains appear to cluster based on region but there were still significant differences within regions. Future research needs to address potential underdiagnosis due to genetic diversity but also to understand mode of transmission, variation in virulence, and the role of host genetics in APPV susceptibility.


Modulation of monocytic lipopolysaccharide-induced tissue factor expression and tumor necrosis factor alpha release by estrogen and calcitriol.
Modulatory effects of estrogens on both the immune and the coagulation system are only partially understood. In severe infections high estrogen levels have been observed both in men and postmenopausal women and are associated with increased mortality. Monocyte-derived tissue factor (TF) expression can activate the coagulation system and worsen the course of severe infection. T he aim of the current study was to evaluate the in vitro effect of estrogens on differentiation, TF expression and Tumor Necrosis Factor alpha (TNFalpha) release in human monocytes. Isolated peripheral blood monocytes, MM6- and T HP-1 cells were cultured and stimulated by lipopolysaccharides (LPS) in the presence of 17beta-estradiol (E2) and/or calcitriol. Proliferative responses were evaluated by determining the proliferation rate and by cell cycle analysis. Cell surface expression of C D14 and T F was determined by flow cytometry. TNFalpha was determined by ELISA. Although calcitriol induced the expression of the differentiation marker C D14 and decreased the expression of T F in both immature monocytic cell lines and primary monocytes, the LPS stimulation of T F expression was not significantly increased in immature monocytic cells and was decreased in mature monocytes. Calcitriol-treatment increased LPS-induced TNFalpha release in MM6 cells but inhibited TNFalpha release from peripheral blood monocytes. Treatment with E2 did not alter the phenotype or cell proliferation of resting monocytic cells. However, E2-treated monocytic cells and monocytes responded to LPS by increased TF expression and decreased TNFalpha. The results suggest that estrogens may modulate T F expression and cytokine production by monocytes and may thus be involved, at least in part, in the pathophysiology of acute inflammatory processes associated with high estrogen levels.


Nuclear translocation of endonuclease G and apoptosis-inducing factor during acetaminophen-induced liver cell injury.
Mitochondrial dysfunction and internucleosomal DNA fragmentation are well-recognized features of acetaminophen (AAP)-induced hepatocyte cell death. However, the endonucleases responsible for this effect have not been identified. Apoptosis-inducing factor (AIF) and endonuclease G are nucleases located in the intermembrane space of mitochondria. AIF is thought to trigger chromatin condensation and induce cleavage of DNA into high molecular weight fragments (50-300 kb), and endonuclease G can produce oligonucleosomal DNA fragments. Therefore, the objective of this investigation was to test the hypothesis that endonuclease G and AIF could be involved in AAP-induced nuclear DNA fragmentation. Using immunofluorescence microscopy, it was shown that in primary cultured mouse hepatocytes, endonuclease G and AIF translocated to the nucleus between 3 and 6 h after exposure to 5 mM AAP. In contrast, other mitochondrial intermembrane proteins such as cytochrome c or the second mitochondria-derived activator of caspases (Smac) did not accumulate in the nucleus. The translocation of AIF and endonuclease G correlated with mitochondrial dysfunction as indicated by the progressive loss of the mitochondrial membrane potential (measured with the JC-1 assay) and the appearance of nuclear DNA fragments in the cytosol (determined by an anti-histone ELISA). Pretreatment with 20mM N-acetylcysteine prevented mitochondrial dysfunction, the nuclear translocation of endonuclease G and AIF, and the nuclear DNA fragmentation. The data support the conclusion that endonuclease G and AIF translocate to the nucleus in response to AAP-induced mitochondrial dysfunction and may be responsible, at least in part, for the initial DNA fragmentation during AAP hepatotoxicity.


DISAPPEARANCE OF A GENETIC MARKER FROM A CYTOPLASMIC HYBRID PLASMODIUM OF A TRUE SLIME MOLD.
Two strains of the true slime mold Didymium nigripes form distinctive plaques when their spores are plated clonally. Strain C6 gives rise to plaques surrounded by large plasmodia. Plaques of strain A20 at first contain only amoebas, the formation of plasmodia being delayed; and strain A20 is resistant to cycloheximide. Plasmodia of C6 and A20 were fused, and at intervals the fused plasmodia were cut into segments and each segment was permitted to fruit. The resulting spores were plated clonally and the plaques scored as C6-type or A20-type. When equal-sized C6 and A20 plasmodia were fused, the A20 characteristic of delayed plasmodium formation disappeared with time so that after 8 hours few A20 plaques could be recovered even from the A20 side of the fused plasmodium. When the ratio between sizes of C6 and A20 plasmodia to be fused was varied, similar results were obtained. The A20 characteristic did not disappear until fruiting.


A Detail-Based Method for Linear Full Reference Image Quality Prediction.
In this paper, a novel full Reference method is proposed for image quality assessment, using the combination of two separate metrics to measure the perceptually distinct impact of detail losses and of spurious details. To this purpose, the gradient of the impaired image is locally decomposed as a predicted version of the original gradient, plus a gradient residual. It is assumed that the detail attenuation identifies the detail loss, whereas the gradient residuals describe the spurious details. It turns out that the perceptual impact of detail losses is roughly linear with the loss of the positional Fisher information, while the perceptual impact of the spurious details is roughly proportional to a logarithmic measure of the signal to residual ratio. The affine combination of these two metrics forms a new index strongly correlated with the empirical differential mean opinion score (DMOS) for a significant class of image impairments, as verified for three independent popular databases. The method allowed alignment and merging of DMOS data coming from these different databases to a common DMOS scale by affine transformations. Unexpectedly, the DMOS scale setting is possible by the analysis of a single image affected by additive noise.


[The changes of physiological reactivity of cardiorespiratory system to respiratory homeostasis with the use of complex stimulation of special work capacity].
We present the influence of the program of special additional stimulation of work capacity of high-performance athletes on the sensitivity of cardiorespiratory system to hypercapnic and hypoxic shifts in respiratory homeostasis. We found that under the influence of the pre-start complex a decrease in the sensitivity of ventilator responses to CO2-H+ stimuli in combination with a reduction in the thresholds of the reaction take place. This creates conditions for increased mobilization properties of the cardiorespiratory system and economization of its reaction under conditions of changes of respiratory homeostasis characteristic of intense training and competitive loads in the sport.


CO2 Absorption in the Ionic Liquids Immobilized on Solid Surface by Molecular Dynamics Simulation.
Based on our previous experimental research, we studied the absorption of CO2 in the ionic liquid, 1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([hmim][Tf2N]), immobilized on TiO2 [rutile (110) ] with different thickness by molecular dynamics simulation. The effects of the properties (hydrophobicity and hydrophilicity) of solid interfaces were also studied with IL immobilized on graphite and TiO2, respectively. We studied the influence of the thickness of IL immobilized on TiO2 on the absorption of CO2 via structural and dynamical properties. The results show that the self-diffusion coefficients of IL and CO2 increase as the thickness of immobilized IL decreases. And the CO2 absorption capacity increases as the thickness of immobilized IL decreases as well. Additionally, more CO2 molecules are absorbed in the region near the solid interface as the thickness of IL decreases. For IL immobilized on graphite, the self-diffusion coefficients of cations and anions are larger than that of IL immobilized on TiO2 with the same thickness. They are also larger than nonimmobilized cations and anions.Besides, the CO2 absorption capacity of IL immobilized on TiO2 is the largest compared with IL immobilized on graphite and nonimmobilized IL with the same thickness. From our simulation work, we try to explore the microscopic mechanism that is unexplored by experimental work, and we found the important role of IL/solid interface for CO2 absorption in immobilized ILs.


Separation of transfer ribonucleic acids on polystyrene anion exchangers.
The transfer RNA separation by chromatography on strong-base polystyrene exchange materials is examined and compared with the widely used reversed-phase chromatography. Results indicate important differences in some transfer RNA (tRNA) elution patterns by the anion-exchange chromatography, as compared with the reversed-phase chromatography. Transfer RNAs containing hydrophobic groups are adsorbed more strongly. The anion exchanger has twice the number of theoretical plates. Single peaks of tRNA2Glu and tRNA1Phe obtained from the reversed-phase column give multiple peaks only polystyrene anion-exchange chromatography. All six leucine tRNAs (Escherichia coli) and differences in tRNA populations synthesized during early and late stages of the dividing lymphocytes from normal human blood can be characterized by the anion-exchange chromatography. Different separation profiles are obtained by two separation systems for tyrosine tRNAs from mouse liver and mouse-plasma-cell tumor. The results indicate that, in contrast to the reversed-phase chromatography, strong-base-polystyrene anion-exchange chromatography is capable of separating tRNAs with minor structlral differences.


Nonlinear Dynamic Analysis of Vowels in Cleft Palate Children with or without Hypernasality.
Objectives. To clarify the difference between Lyapunov exponents (LEs) for cleft palate (CP) patients with hypernasality versus without hypernasality and to investigate the relationship between their LEs and nasalance scores (NSs). Material and Methods. Six CP patients with severe hypernasality (mean age 9.2 years) and six CP patients without hypernasality (mean age 8.0 years) were enrolled. Five Japanese vowels were recorded at 44.1 KHz, and the NSs were measured simultaneously. The mean first LE (mLE(1)) from all one-second intervals was computed. Results. The mLE(1) for /o/ in patients with hypernasality was significantly higher than that in patients without hypernasality. The correlation coefficients between the mLE(1) and NS for all vowels were not statistically different. Conclusion. The voice signal of /o/ for the patients with hypernasality was more instable than in those without hypernasality. The chaotic phenomenon was independent of nasal resonance in CP voice.


Texture Features of Proton Density Fat Fraction Maps from Chemical Shift Encoding-Based MRI Predict Paraspinal Muscle Strength.
Texture analysis (TA) has shown promise as a surrogate marker for tissue structure, based on conventional and quantitative MRI sequences. Chemical-shift-encoding-based MRI (CSE-MRI)-derived proton density fat fraction (PDFF) of paraspinal muscles has been associated with various medical conditions including lumbar back pain (LBP) and neuromuscular diseases (NMD). Its application has been shown to improve the prediction of paraspinal muscle strength beyond muscle volume. Since mean PDFF values do not fully reflect muscle tissue structure, the purpose of our study was to investigate PDFF-based TA of paraspinal muscles as a predictor of muscle strength, as compared to mean PDFF. We performed 3T-MRI of the lumbar spine in 26 healthy subjects (age = 30 ± 6 years; 15 females) using a six-echo 3D spoiled gradient echo sequence for chemical-shift-encoding-based water-fat separation. Erector spinae (ES) and psoas (PS) muscles were segmented bilaterally from level L2-L5 to extract mean PDFF and texture features. Muscle flexion and extension strength was measured with an isokinetic dynamometer. Out of the eleven texture features extracted for each muscle, Kurtosis(global) of ES showed the highest significant correlation (r = 0.59, p = 0.001) with extension strength and Variance(global) of PS showed the highest significant correlation (r = 0.63, p = 0.001) with flexion strength. Using multivariate linear regression models, Kurtosis(global) of ES and BMI were identified as significant predictors of extension strength (R2adj = 0.42; p < 0.001), and Variance(global) and Skewness(global) of PS were identified as significant predictors of flexion strength (R2adj = 0.59; p = 0.001), while mean PDFF was not identified as a significant predictor. TA of CSE-MRI-based PDFF maps improves the prediction of paraspinal muscle strength beyond mean PDFF, potentially reflecting the ability to quantify the pattern of muscular fat infiltration. In the future, this may help to improve the pathophysiological understanding, diagnosis, monitoring and treatment evaluation of diseases with paraspinal muscle involvement, e.g., NMD and LBP.


Predicting Drug-Target Interactions With Multi-Information Fusion.
Identifying potential associations between drugs and targets is a critical prerequisite for modern drug discovery and repurposing. However, predicting these associations is difficult because of the limitations of existing computational methods. Most models only consider chemical structures and protein sequences, and other models are oversimplified. Moreover, datasets used for analysis contain only true-positive interactions, and experimentally validated negative samples are unavailable. To overcome these limitations, we developed a semi-supervised based learning framework called NormMulInf through collaborative filtering theory by using labeled and unlabeled interaction information. The proposed method initially determines similarity measures, such as similarities among samples and local correlations among the labels of the samples, by integrating biological information. The similarity information is then integrated into a robust principal component analysis model, which is solved using augmented Lagrange multipliers. Experimental results on four classes of drug-target interaction networks suggest that the proposed approach can accurately classify and predict drug-target interactions. Part of the predicted interactions are reported in public databases. The proposed method can also predict possible targets for new drugs and can be used to determine whether atropine may interact with alpha1B- and beta1- adrenergic receptors. Furthermore, the developed technique identifies potential drugs for new targets and can be used to assess whether olanzapine and propiomazine may target 5HT2B. Finally, the proposed method can potentially address limitations on studies of multitarget drugs and multidrug targets.


Increased muscle size and strength from slow-movement, low-intensity resistance exercise and tonic force generation.
The authors investigated the effects of low-intensity resistance training on muscle size and strength in older men and women. Thirty-five participants (age 59-76 yr) were randomly assigned to 2 groups and performed low-intensity (50% of 1-repetition maximum) knee-extension and -flexion exercises with either slow movement and tonic force generation (LST; 3-s eccentric, 3-s concentric, and 1-s isometric actions with no rest between repetitions) or normal speed (LN; 1-s concentric and 1-s eccentric actions with 1-s rests between repetitions) twice a week for 12 wk (2-wk preparation and 10-wk intervention). The LST significantly increased thigh-muscle thickness, as well as isometric knee-extension and -flexion strength. The LN significantly improved strength, but its hypertrophic effect was limited. These results indicate that even for older individuals, the LST can be an effective method for gaining muscle mass and strength.


An objective method for the production of isopach maps and implications for the estimation of tephra deposit volumes and their uncertainties.
Characterization of explosive volcanic eruptive processes from interpretation of deposits is a key for assessing volcanic hazard and risk, particularly for infrequent large explosive eruptions and those whose deposits are transient in the geological record. While eruption size-determined by measurement and interpretation of tephra fall deposits-is of particular importance, uncertainties for such measurements and volume estimates are rarely presented. Here, tephra volume estimates are derived from isopach maps produced by modeling raw thickness data as cubic B-spline curves under tension. Isopachs are objectively determined in relation to original data and enable limitations in volume estimates from published maps to be investigated. The eruption volumes derived using spline isopachs differ from selected published estimates by 15-40 %, reflecting uncertainties in the volume estimation process. The formalized analysis enables identification of sources of uncertainty; eruptive volume uncertainties (>30 %) are much greater than thickness measurement uncertainties (~10 %). The number of measurements is a key factor in volume estimate uncertainty, regardless of method utilized for isopach production. Deposits processed using the cubic B-spline method are well described by 60 measurements distributed across each deposit; however, this figure is deposit and distribution dependent, increasing for geometrically complex deposits, such as those exhibiting bilobate dispersion.


Ratiometric optical sensor for high-resolution imaging of pH with low cross-talk.
We synthesize lipophilic, highly efficient, and pH-insensitive oleic acid-modified quantum dots (QDs) with maximum emission at a wavelength of 628 nm. The pH sensing film is fabricated by encapsulating 5-hexadecanoylamino-fluorescein and QDs as the reference in D4-hydromed and plasticized polystyrene. Using a light-emitting diode with a central wavelength of 410 nm as an excitation source, it is shown that the emission wavelengths of the pH sensitive indicator and reference dye have no spectral overlap and match respectively the channels of a 3CCD (RGB) camera with low cross-talk. A series of validation experiments shows that this ratiometric pH optode has good properties of high sensitivity, long-term stability, and photostability. It had a fast response time of <20 s when going from pH 6.3 to pH 8.0. The pH images suggest that the proposed ratiometric pH-sensing approach has great advantage and promise for field applications.


Home Parenteral Nutrition: Vascular Access and Related Complications.
Patients with chronic intestinal failure are dependent on parenteral nutrition (PN) to maintain health and preserve life. Maintaining safe vascular access is vital to prevent life-threatening complications such as catheter-related bloodstream infection or central venous occlusion. Dedicated central venous catheters with rigorous catheter care aseptic protocols are vital in obtaining good long-term outcomes that allow continuation of PN over many years. Good catheter care requires an experienced multidisciplinary team using appropriate vascular devices, trained to identify and aggressively treat catheter-related bloodstream infections, catheter occlusions, and catheter-related thrombosis. Consideration must also be given to evolving strategies to prevent recurrent infections, including prophylactic central venous catheter locks.


Blood Metabolomic Predictors of 1-Year Outcome in Subarachnoid Hemorrhage.
Delayed neurological deficit (DND) is the most important cause of morbidity and mortality in patients with subarachnoid hemorrhage (SAH) whose aneurysms have been secured. However, the methods currently used to predict the development of DND, such as trans-cranial Doppler or levels biochemical markers in blood and cerebrospinal fluid are not very accurate. Venous blood was drawn from 50 patients with SAH, admitted to the neurosurgical department Umeå University Hospital, at day 1-3 and day 7 after the bleed. The clinical status of the patients was followed up approximately 1 year after this episode and classified according to the Glasgow Outcome Score (GOS). Results showed considerable differences in blood metabolomic patterns between day 1-3 and 7 after the hemorrhage. Fifty-six out of 98 metabolites could be identified from our in-house library and 17 of these metabolites changed significantly from day 1-3 to 7 after the bleed. One of these, myo-inositol, was predictive of clinical outcome even after correction for multiple testing. An estimation of the diagnostic accuracy of high levels of this substance in predicting good outcome (GOS 4-5) yielded a sensitivity of .763 and a specificity of .5 at the optimal cut off point. SAH is an event with a profound effect on blood metabolomics profiles. Myo-inositol might be an interesting compound for future study to focus on in the search for metabolic markers in venous blood of delayed neurological deterioration in SAH patients.


Lophodermium macci sp. nov., a new species on senesced foliage of five-needle pines.
The new species Lophodermium macci is described. It is similar in its morphology, habitat, geographic range and ecology to L. pini-excelsae, L. staleyi and L. nitens and often is misidentified as L. pinastri on Pinus strobus in herbaria. A modified technique was used to extract DNA from minute ascomata on herbarium specimens, and new primers were made to amplify the damaged DNA from these specimens. It provides added evidence to separate L. macci from L. pini-excelsae, its closest morphological taxon.


The association between strenuous physical activity and obesity in a high and low altitude populations in southern Saudi Arabia.
To assess the relationship between obesity and strenuous physical activity in Saudi nationals living at high and low altitude. Cross-sectional randomized study. A total of 437 healthy adult Saudi nationals born and living permanently at high altitude (3150 m) and 468 healthy adult Saudi nationals born and living permanently at relatively low altitude 500 m). Body Mass Index (Weight (kg)/height (m2)) strenuous activity scores using Lipid Research Clinic Questionnaire. Resting radial pulse rate (beats/min). Strenuous physical activity was significantly and inversely associated with obesity in men at both high (chi 2 = 7.13, P < 0.05) and low (chi 2 = 6.14, P < 0.05) altitude but there was no clear trend for women at either altitude. The lack of association between strenuous physical activity and obesity in women was attributed to the low and homogeneous levels of strenuous physical activity. Strenuous physical activity should be encouraged as strategy directed towards weight reduction in obese as well as prevention of obesity in Saudi high and lowlanders.


Cortical disinhibition occurs in chronic neuropathic, but not in chronic nociceptive pain.
The aim of this study was to examine the relationship between chronic neuropathic pain after incomplete peripheral nerve lesion, chronic nociceptive pain due to osteoarthritis, and the excitability of the motor cortex assessed by transcranial magnetic stimulation (TMS). Hence in 26 patients with neuropathic pain resulting from an isolated incomplete lesion of the median or ulnar nerve (neuralgia), 20 patients with painful osteoarthritis of the hand, and 14 healthy control subjects, the excitability of the motor cortex was tested using paired-pulse TMS to assess intracortical inhibition and facilitation. These excitability parameters were compared between groups, and the relationship between excitability parameters and clinical parameters was examined. We found a significant reduction of intracortical inhibition in the hemisphere contralateral to the lesioned nerve in the neuralgia patients. Intracortical inhibition in the ipsilateral hemisphere of neuralgia patients and in both hemispheres of osteoarthritis patients did not significantly differ from the control group. Disinhibition was significantly more pronounced in neuralgia patients with moderate/severe pain intensity than in patients with mild pain intensity, whereas the relative compound motor action potential as a parameter of nerve injury severity did not correlate with the amount of disinhibition. Our results suggest a close relationship between motor cortex inhibition and chronic neuropathic pain in the neuralgia patients, which is independent from nerve injury severity. The lack of cortical disinhibition in patients with painful osteoarthritis points at differences in the pathophysiological processes of different chronic pain conditions with respect to the involvement of different brain circuitry.


Inactivation of the adrenergic receptor β2 disrupts glucose homeostasis in mice.
Three types of beta adrenergic receptors (ARβ1-3) mediate the sympathetic activation of brown adipose tissue (BAT), the key thermogenic site for mice which is also present in adult humans. In this study, we evaluated adaptive thermogenesis and metabolic profile of a mouse with Arβ2 knockout (ARβ2KO). At room temperature, ARβ2KO mice have normal core temperature and, upon acute cold exposure (4 °C for 4 h), ARβ2KO mice accelerate energy expenditure normally and attempt to maintain body temperature. ARβ2KO mice also exhibited normal interscapular BAT thermal profiles during a 30-min infusion of norepinephrine or dobutamine, possibly due to marked elevation of interscapular BAT (iBAT) and of Arβ1, and Arβ3 mRNA levels. In addition, ARβ2KO mice exhibit similar body weight, adiposity, fasting plasma glucose, cholesterol, and triglycerides when compared with WT controls, but exhibit marked fasting hyperinsulinemia and elevation in hepatic Pepck (Pck1) mRNA levels. The animals were fed a high-fat diet (40% fat) for 6 weeks, ARβ2KO mice doubled their caloric intake, accelerated energy expenditure, and induced Ucp1 expression in a manner similar to WT controls, exhibiting a similar body weight gain and increase in the size of white adipocytes to the WT controls. However, ARβ2KO mice maintain fasting hyperglycemia as compared with WT controls despite very elevated insulin levels, but similar degrees of liver steatosis and hyperlipidemia. In conclusion, inactivation of the ARβ2KO pathway preserves cold- and diet-induced adaptive thermogenesis but disrupts glucose homeostasis possibly by accelerating hepatic glucose production and insulin secretion. Feeding on a high-fat diet worsens the metabolic imbalance, with significant fasting hyperglycemia but similar liver structure and lipid profile to the WT controls.


[Chemical protection of golden hamsters against ionizing radiation].
It was shown on Golden hamsters, which are characterized by high radioresistance of the intestine, that S-2-(omega-aminopropylaminoethyl) thiophosphorous acid (gammaphos) exerts a protective action against both X- and neutron-radiation although in the latter case the protection is less pronounced. In conditions of hibernation, the protective effect of gammaphos against X-radiation was not statistically reliable. Hibernation during exposure and at the postirradiation period increases considerably the radioresistance of animals. The influence of hibernation depends upon its duration.


Topical niacin cream-assisted 595-nm pulsed-dye laser treatment for facial flushing: retrospective analysis of 25 Korean patients.
Flushing is defined clinically as a transient reddening of the face and other areas. Due to the transient nature of flushing, a patient may not show signs of flushing during laser treatment. The aim of this study was to evaluate the efficacy and safety of 595-nm pulsed-dye laser treatment of flushing or erythema after provocation of flushing by topical niacin cream. We retrospectively reviewed a total of 25 Korean patients with facial flushing who were treated with three sessions of 595-nm pulsed-dye laser after the application of topical niacin cream. Follow-up results revealed that 12 of the 25 patients demonstrated marked (51-75%) clinical improvement of baseline facial erythema. Eight patients had moderate (26-50%) improvement and three demonstrated near total (≥ 75%) improvement. Two patients showed minimal to no (0-25%) improvement. We observed that the reactivity to topical niacin cream was markedly reduced in 64% of our patients after 595-nm pulsed-dye laser treatments. Minimal post-therapy facial oedema was noted in most of the patients, which usually resolved spontaneously within 2 days. Pronounced facial swelling was observed in four patients. We suggest that 595-nm pulsed-dye laser treatment after provocation of flushing by topical niacin cream may provide a new treatment algorithm for facial flushing in Asians.


Stenotrophomonas, Achromobacter, and nonmelioid Burkholderia species: antimicrobial resistance and therapeutic strategies.
Stenotrophomonas maltophilia, Achromobacter xylosoxidans, and nonmelioid Burkholderia species, namely, Burkholderia cepacia complex, collectively are a group of troublesome nonfermenters. Although not inherently virulent organisms, these environmental Gram negatives can complicate treatment in those who are immunocompromised, critically ill in the intensive care unit and those patients with suppurative lung disease, such as cystic fibrosis. Through a range of intrinsic antimicrobial resistance mechanisms, virulence factors, and the ability to survive in biofilms, these opportunistic pathogens are well suited to persist, both in the environment and the host. Treatment recommendations are hindered by the difficulties in laboratory identification, the lack of reproducibility of antimicrobial susceptibility testing, the lack of clinical breakpoints, and the absence of clinical outcome data. Despite trimethoprim-sulfamethoxazole often being the mainstay of treatment, resistance is widely encountered, and alternative regimens, including combination therapy, are often used. This review will highlight the important aspects and unique challenges that these three nonfermenters pose, and, in the absence of clinical outcome data, our therapeutic recommendations will be based on reported antimicrobial susceptibility and pharmacokinetic/pharmacodynamic profiles.


E-learning as an Effective Method in the Prevention of Patient Falls.
Patient falls deteriorate patients' functional condition and quality of life, and increase their treatment costs. E-learning is considered an effective way to gain knowledge and competencies for quality and safety in nursing practice. The aim of the study is to evaluate the effectiveness of an e-learning course for nurses in preventing in-patient falls. The research design was mixed. In the first phase of the study, a five-year retrospective analysis of 2,280 in-patient falls was performed. Based on the analysis of risk factors for patient falls and group interviews with clinic managers an e-learning course was designed and completed by 250 nurses from five surgical and internal departments. The course's effectiveness was evaluated based on the incidence of patient falls and the consequences of the falls before and after e-learning. At surgical departments, there was a statistically significant decrease in patient fall indices after the implementation of the e-learning course (from 4.4 to 2.6 falls per 1,000 patients; p=0.022). On the contrary, in internal departments, this index increased in the monitored period (from 19.0 to 26.9 falls per 1,000 patients; p=0.001). In all departments, there was a decrease in the incidence of patient injuries caused by falls after the implementation of the e-learning course; in internal medicine, this decrease was statistically significant (from 54.5% to 33.3%; p=0.014). The study confirmed that e-learning forms of education for healthcare professionals have a positive effect in preventing patient falls.


Ultrasound Surface Wave Elastography for Assessing Scleroderma.
Scleroderma, or systemic sclerosis (SSc), is a multi-organ connective tissue disease characterized by immune dysregulation and tissue fibrosis. Skin disease is both a disabling feature of SSc and a predictor of visceral involvement and increased mortality. The Modified Rodnan Skin Score (MRSS) is currently the most common clinical method for assessing skin. We developed ultrasound surface wave elastography (USWE) techniques to measure skin surface wave speeds and analyze skin viscoelasticity. The objective of this research was to determine the correlations of skin surface wave speed and skin viscoelasticity with MRSS. Twenty-six SSc patients were studied using USWE and the MRSS. The subject was tested in a sitting position while his or her left or right forearm and upper arm were placed horizontally on a pillow in a relaxed state. The skin of both left and right forearms and upper arms of patients was tested using USWE. Surface wave speeds are positively correlated with the MRSS. Skin elasticity is also positively correlated with the MRSS. However, there was no correlation between skin viscosity and the MRSS for these SSc patients. We will further study if skin viscosity is sensitive enough to detect early edema from inflammation changes of SSc.


Changing dental caries and periodontal disease patterns among a cohort of Ethiopian immigrants to Israel: 1999-2005.
Dental epidemiology has indicated that immigrants and minority ethnic groups should be regarded as high risk populations on the verge of oral health deterioration. The objectives of this study were to measure the changing pattern of dental caries, periodontal health status and tooth cleaning behaviour among a cohort of Ethiopian immigrants to Israel between the years 1999-2005. Increment of dental caries and periodontal health status was recorded among a cohort of 672 Ethiopian immigrants, utilizing the DMFT and CPI indices. Data were gathered during 1999-2000 and five years later, during 2004-2005. Participants were asked about their oral hygiene habits in Ethiopia and in Israel five years since their immigration. Regarding dental caries, at baseline 70.1% of the examinees were caries-free, as compared to 57.3% after five years. DMFT had increased from 1.48 to 2.31. For periodontal health status, at baseline, 94.7% demonstrated no periodontal pockets (CPI scores 0-2) and 5.3% revealed periodontal pockets (CPI scores 3&4), compared to 75.6% and 24.4%, respectively after five years. At baseline, 74% reported cleaning their teeth exclusively utilizing chewing and cleaning sticks common in Ethiopia. After five years, 97% reported cleaning their teeth exclusively utilizing toothbrushes. The deterioration in the oral health status, especially the alarming and significant worsening of periodontal health status, among this immigrant group, emphasizes the need for health promotion and maintenance among immigrants and minority groups in changing societies. An "acclimatizing and integrating" model of oral health promotion among minority and immigrant groups is suggested.


Lack of Diagnostic Pluripotentiality in Patients at Clinical High Risk for Psychosis: Specificity of Comorbidity Persistence and Search for Pluripotential Subgroups.
More than 20 years after the clinical high risk syndrome for psychosis (CHR) was first articulated, it remains controversial whether the CHR syndrome predicts onset of psychosis with diagnostic specificity or predicts pluripotential diagnostic outcomes. Recently, analyses of observational studies, however, have suggested that the CHR syndrome is not pluripotential for emergent diagnostic outcomes. The present report conducted additional analyses in previously reported samples to determine (1) whether comorbid disorders were more likely to persist in CHR patients compared to a comparison group of patients who responded to CHR recruitment efforts but did not meet criteria, termed help-seeking comparison subjects (HSC); and (2) whether clinically defined pluripotential CHR subgroups could be identified. All data were derived from 2 multisite studies in which DSM-IV structured diagnostic interviews were conducted at baseline and at 6-month intervals. Across samples we observed persistence of any nonpsychotic disorder in 80/147 CHR cases (54.4%) and in 48/84 HSC cases (57.1%, n.s.). Findings with persistence of anxiety, depressive, and bipolar disorders considered separately were similar. Efforts to discover pluripotential CHR subgroups were unsuccessful. These findings add additional support to the view that the CHR syndrome is not pluripotential for predicting various diagnostic outcomes but rather is specific for predicting emergent psychosis.


Wait times for gastroenterology consultation in Canada: the patients' perspective.
Long wait times for health care have become a significant issue in Canada. As part of the Canadian Association of Gastroenterology's Human Resource initiative, a questionnaire was developed to survey patients regarding wait times for initial gastroenterology consultation and its impact. A total of 916 patients in six cities from across Canada completed the questionnaire at the time of initial consultation. Self-reported wait times varied widely, with 26.8% of respondents reporting waiting less than two weeks, 52.4% less than one month, 77.1% less than three months, 12.5% reported waiting longer than six months and 3.6% longer than one year. One-third of patients believed their wait time was too long, with 9% rating their wait time as 'far too long'; 96.4% believed that maximal wait time should be less than three months, 78.9% believed it should be less than one month and 40.3% believed it should be less than two weeks. Of those working or attending school, 22.6% reported missing at least one day of work or school because of their symptoms in the month before their appointment, and 9.0% reported missing five or more days in the preceding month. A total of 20.2% of respondents reported being very worried about having a serious disease (ie, scored 6 or higher on 7-point Likert scale), and 17.6% and 14.8%, respectively, reported that their symptoms caused major impairment of social functioning and with the activities of daily living. These data suggest that a significant proportion of Canadians with digestive problems are not satisfied with their wait time for gastroenterology consultation. Furthermore, while awaiting consultation, many patients experience an impaired quality of life because of their gastrointestinal symptoms.


A novel surgical approach for the treatment of tumors in the lesser trochanter.
The proximal femur is a common location for the development of primary benign bone tumors. However, there is currently no surgical technique designed specifically for treating tumors located in the lesser trochanter. In the present study, a novel procedure was developed for directly exposing the lesser trochanter for surgical intervention. This technique may be particularly suited to treating tumors that extend toward the lateral and anterior forward of the lesser trochanter. The new approach involved passing through the femoral triangle, separating the femoral nerve and femoral vessels (artery and vein) and resecting the tumor between the iliopsoas and pectineal muscles. The procedure was performed on six patients with various types of tumor, including one case with osteoid osteoma, one case with non-osteogenic fibroma, one case with osteoma, one case with liposarcoma and two cases of osteochondroma. The preliminary results indicated that the surgical durations were short (60-100 min), blood loss was minimal (30-200 ml) and that pain relief was achieved following surgery. Only one patient continued to experience mild pain, scoring 18 mm on a visual analog scale. The other patients were fully relieved of pain. Sensory dysfunction was experienced by one patient following surgery, with persistent numbness and paresthesias in the distribution of the femoral nerve. No cases of deep vein thrombosis, femur head necrosis, hip joint degeneration disease or local recurrence were identified in any patients during the follow-up period. In order to clarify the virtual tissue, such vessels, nerves and the available space in our approach area, we collected 20 cadaveric specimens and performed anatomical examinations in and around the formal triangle. The spaces between the femoral artery and femoral nerve were measured and analyzed, with the results demonstrating that a definite space existed. Therefore, the novel approach presented in the study may be useful in the resection of benign tumors and the preoperative palliative resection of malignant tumors. The technique may be particularly suited to tumors extending toward the lateral and anterior of the lesser trochanter.


Large-scale network interactions supporting item-context memory formation.
Episodic memory is thought to involve functional interactions of large-scale brain networks that dynamically reconfigure depending on task demands. Although the hippocampus and closely related structures have been implicated, little is known regarding how large-scale and distributed networks support different memory formation demands. We investigated patterns of interactions among distributed networks while human individuals formed item-context memories for two stimulus categories. Subjects studied object-scene and object-location associations in different fMRI sessions. Stimulus-responsive brain regions were organized based on their fMRI interconnectivity into networks and modules using probabilistic module-detection algorithms to maximize measurement of individual differences in modular structure. Although there was a great deal of consistency in the modular structure between object-scene and object-location memory formation, there were also significant differences. Interactions among functional modules predicted later memory accuracy, explaining substantial portions of variability in memory formation success. Increased interactivity of modules associated with internal thought and anti-correlation of these modules with those related to stimulus-evoked processing robustly predicted object-scene memory, whereas decreased interactivity of stimulus-evoked processing modules predicted object-location memory. Assessment of individual differences in network organization therefore allowed identification of distinct patterns of functional interactions that robustly predicted memory formation. This highlights large-scale brain network interactions for memory formation and indicates that although networks are largely robust to task demands, reconfiguration nonetheless occurs to support distinct memory formation demands.


Nano-sized Mo- and Nb-doped TiO2 as anode materials for high energy and high power hybrid Li-ion capacitors.
Nano-sized Mo-doped titania (Mo0.1Ti0.9O2) and Nb-doped titania (Nb0.25Ti0.75O2) were directly synthesized via a continuous hydrothermal flow synthesis process. Materials characterization was conducted using physical techniques such as transmission electron microscopy, powder x-ray diffraction, x-ray photoelectron spectroscopy, Brunauer-Emmett-Teller specific surface area measurements and energy dispersive x-ray spectroscopy. Hybrid Li-ion supercapacitors were made with either a Mo-doped or Nb-doped TiO2 negative electrode material and an activated carbon (AC) positive electrode. Cells were evaluated using electrochemical testing (cyclic voltammetry, constant charge discharge cycling). The hybrid Li-ion capacitors showed good energy densities at moderate power densities. When cycled in the potential window 0.5-3.0 V, the Mo0.1Ti0.9O2/AC hybrid supercapacitor showed the highest energy densities of 51 Wh kg-1 at a power of 180 W kg-1 with energy densities rapidly declining with increasing applied specific current. In comparison, the Nb0.25Ti0.75O2/AC hybrid supercapacitor maintained its energy density of 45 Wh kg-1 at 180 W kg-1 better, showing 36 Wh g-1 at 3200 W kg-1, which is a very promising mix of high energy and power densities. Reducing the voltage window to the range 1.0-3.0 V led to an increase in power density, with the Mo0.1Ti0.9O2/AC hybrid supercapacitor giving energy densities of 12 Wh kg-1 and 2.5 Wh kg-1 at power densities of 6700 W kg-1 and 14 000 W kg-1, respectively.


Medical Tourism, Medical Migration, and Global Justice: Implications for Biosecurity in a Globalized World.
We live in the age of globalization. In medicine, that globalization has brought many benefits such as the diffusion of technology and the spread of health care training, but it has also brought threats to biosecurity. This article examines how medical tourism and medical migration pose risks to biosecurity. It also argues that designing legal responses to these risks requires not only technical competence but also a theory of global justice to guide that design.


Retinal Development and Pathophysiology in Kcnj13 Knockout Mice.
Purpose: We constructed and characterized knockout and conditional knockout mice for KCNJ13, encoding the inwardly rectifying K+ channel of the Kir superfamily Kir7.1, mutations in which cause both Snowflake Vitreoretinal Degeneration (SVD) and Retinitis pigmentosa (RP) to further elucidate the pathology of this disease and to develop a potential model system for gene therapy trials. Methods: A Kcnj13 knockout mouse line was constructed by inserting a gene trap cassette expressing beta-galactosidase flanked by FRT sites in intron 1 with LoxP sites flanking exon two and converted to a conditional knockout by FLP recombination followed by crossing with C57BL/6J mice having Cre driven by the VMD2 promoter. Lentiviral replacement of Kcnj13 was driven by the EF1a or VMD2 promoters. Results: Blue-Gal expression is evident in E12.5 brain ventricular choroid plexus, lens, neural retina layer, and anterior RPE. In the adult eye expression is seen in the ciliary body, RPE and choroid. Adult conditional Kcnj13 ko mice show loss of photoreceptors in the outer nuclear layer, inner nuclear layer thinning with loss of bipolar cells, and thinning and disruption of the outer plexiform layer, correlating with Cre expression in the overlying RPE which, although preserved, shows morphological disruption. Fundoscopy and OCT show signs of retinal degeneration consistent with the histology, and photopic and scotopic ERGs are decreased in amplitude or extinguished. Lentiviral based replacement of Kcnj13 resulted in increased ERG c- but not a- or b- wave amplitudes. Conclusion: Ocular KCNJ13 expression starts in the choroid, lens, ciliary body, and anterior retina, while later expression centers on the RPE with no/lower expression in the neuroretina. Although KCNJ13 expression is not required for survival of the RPE, it is necessary for RPE maintenance of the photoreceptors, and loss of the photoreceptor, outer plexiform, and outer nuclear layers occur in adult KCNJ13 cKO mice, concomitant with decreased amplitude and eventual extinguishing of the ERG and signs of retinitis pigmentosa on fundoscopy and OCT. Kcnj13 replacement resulting in recovery of the ERG c- but not a- and b-waves is consistent with the degree of photoreceptor degeneration seen on histology.


Advances in myelofibrosis: a clinical case approach.
Primary myelofibrosis is a member of the myeloproliferative neoplasms, a diverse group of bone marrow malignancies. Symptoms of myelofibrosis, particularly those associated with splenomegaly (abdominal distention and pain, early satiety, dyspnea, and diarrhea) and constitutional symptoms, represent a substantial burden to patients. Most patients eventually die from the disease, with a median survival ranging from approximately 5-7 years. Mutations in Janus kinase 2 (JAK2), a kinase that is essential for the normal development of erythrocytes, granulocytes, and platelets, notably the V617F mutation, have been identified in approximately 50% of patients with myelofibrosis. The approval of a JAK2 inhibitor in 2011 has improved the outlook of many patients with myelofibrosis and has changed the treatment landscape. This article focuses on some of the important issues in current myelofibrosis treatment management, including differentiation of myelofibrosis from essential thrombocythemia and polycythemia vera, up-dated data on the results of JAK2 inhibitor therapy, the role of epigenetic mechanisms in myelofibrosis pathogenesis, investigational therapies for myelofibrosis, and advances in hematopoietic stem cell transplant. Three myelofibrosis cases are included to underscore the issues in diagnosing and treating this complex disease.


Characterization of a mycoplasma virus (MV-O1) derived from and infecting Acholeplasma oculi.
A new Mycoplasmatales virus, referred to as MV-O1, was isolated during cloning of Acholeplasma oculi 19L. The virus formed plaques only on strains of A. oculi, i.e. the original clone, A. oculi 19L, a subclone of A. oculi 19L (A.oculi-i), A. oculi Goat 5 and wild isolate (K-2) of A. oculi, but not on other acholeplasmas, including strains of A. laidlawii, nor on five human mycoplasma species tested. The virus required horse serum for multiplication as well as for plaque formation and passed through a 100 nm filter. Electron microscopy revealed enveloped, spherical particles 80-130 nm in diameter. The buoyant density of purified virus was 1.23 g ml-1 in CsCl, and agarose gel electrophoresis indicated that the viral nucleic acid was DNA.


On the fidelity of DNA synthesis. Pyrophosphate-induced misincorporation allows detection of two proofreading mechanisms.
The effect of pyrophosphate on the fidelity of in vitro DNA synthesis has been examined. Pyrophosphate enhances misincorporation by Escherichia coli DNA polymerase I in copying phi X174 DNA. The increased misincorporation is directly proportional to the extent of inhibition of the rate of polymerization. In contrast, pyrophosphate is not detectably mutagenic with avian myeloblastosis virus DNA polymerase or DNA polymerases alpha and beta from animal cells, which lack associated proofreading activities. This suggests that increased misincorporation by pyrophosphate is not due to an increase in misinsertions by DNA polymerase, but rather due to inhibition of proofreading by pyrophosphate. However, the pyrophosphate-induced infidelity has a different specificity from, and is not competitive with, two experimental markers of 3'----5' exonuclease proofreading; i.e. the effects of the next nucleotide or the addition of deoxynucleoside monophosphates. These distinctive features suggest a second mode of proofreading susceptible to inhibition by pyrophosphate. This concept is discussed in relation to models for proofreading described in the literature.


Personalized metabolomics for predicting glucose tolerance changes in sedentary women after high-intensity interval training.
High-intensity interval training (HIIT) offers a practical approach for enhancing cardiorespiratory fitness, however its role in improving glucose regulation among sedentary yet normoglycemic women remains unclear. Herein, multi-segment injection capillary electrophoresis-mass spectrometry is used as a high-throughput platform in metabolomics to assess dynamic responses of overweight/obese women (BMI > 25, n = 11) to standardized oral glucose tolerance tests (OGTTs) performed before and after a 6-week HIIT intervention. Various statistical methods were used to classify plasma metabolic signatures associated with post-prandial glucose and/or training status when using a repeated measures/cross-over study design. Branched-chain/aromatic amino acids and other intermediates of urea cycle and carnitine metabolism decreased over time in plasma after oral glucose loading. Adaptive exercise-induced changes to plasma thiol redox and orthinine status were measured for trained subjects while at rest in a fasting state. A multi-linear regression model was developed to predict changes in glucose tolerance based on a panel of plasma metabolites measured for naïve subjects in their untrained state. Since treatment outcomes to physical activity are variable between-subjects, prognostic markers offer a novel approach to screen for potential negative responders while designing lifestyle modifications that maximize the salutary benefits of exercise for diabetes prevention on an individual level.


Enhanced cardiac function in transgenic mice expressing a Ca(2+)-stimulated adenylyl cyclase.
The predominant functional adenylyl cyclases normally expressed in cardiac tissue and coupled to beta-adrenergic receptors are inhibited by micromolar Ca(2+) concentration. To modify the overall balance of activities, we have generated transgenic mice expressing the Ca(2+)-stimulatable adenylyl cyclase type 8 (AC8) specifically in the heart. AC activity is increased by at least 7-fold in heart membranes from transgenic animals and is stimulated by Ca(2+) in the same range of concentration that inhibits the endogenous activity. Moreover, the in vivo basal protein kinase A activity was augmented 4-fold. Overexpression of AC8 in the heart has no detrimental consequences on global cardiac function. Basal heart rate and contractile function, measured by noninvasive echocardiography, were unchanged. In contrast, on release of parasympathetic tone, the intrinsic contractility is heightened and unresponsive to further beta-adrenergic receptor stimulation. AC8 transgenic mice thus represent an original model to investigate the relative influence of Ca(2+) and cAMP on cardiac function within a phenotype of enhanced cardiac contractility and relaxation.


Descriptive study of patients receiving excision and radiotherapy for keloids.
To review and describe our institution's outcomes in patients treated with external beam radiotherapy after keloid excision. This was a retrospective study. Patients who received radiotherapy between July 1994 and January 2004 after keloid excision were identified. A questionnaire was mailed regarding sociodemographic factors, early and late radiation toxicities, the need for additional therapy, and satisfaction level. All patients had received a total of 15 Gy in three daily 5-Gy fractions. Treatment started within 24 h after surgery and was delivered on a Siemens orthovoltage machine. The data were analyzed using the STATA statistical package. A total of 234 patients were approached. The response rate was 41%, and 75% were female. The mean age was 36.5 years (range, 16-69 years). The patients were mainly of European (53.1%) or African (19.8%) descent. For early toxicity outcomes, 54.2% reported skin redness and 24% reported skin peeling. For late toxicity outcomes, 27% reported telangiectasia and 62% reported permanent skin color changes. No association was found with gender, skin color, or age for the late toxicity outcomes. Of the patients responding, 14.6% required adjuvant treatment. On a visual scale of 1-10 for the satisfaction level, 60% reported a satisfaction level of > or =8. Telangiectasia was the most significant predictor of a low satisfaction level (< or =3, p < 0.005). The results of our study have shown that orthovoltage-based radiotherapy after surgical excision for keloids is a good method for the prevention of relapse. It is well tolerated, causes little toxicity, and leads to a high patient satisfaction level.


A case-control study on the association between bladder cancer and prior bladder calculus.
Bladder calculus is associated with chronic irritation and inflammation. As there is substantial documentation that inflammation can play a direct role in carcinogenesis, to date the relationship between stone formation and bladder cancer (BC) remains unclear. This study aimed to examine the association between BC and prior bladder calculus using a population-based dataset. This case-control study included 2,086 cases who had received their first-time diagnosis of BC between 2001 and 2009 and 10,430 randomly selected controls without BC. Conditional logistic regressions were employed to explore the association between BC and having been previously diagnosed with bladder calculus. Of the sampled subjects, bladder calculus was found in 71 (3.4%) cases and 105 (1.1%) controls. Conditional logistic regression analysis revealed that the odds ratio (OR) of having been diagnosed with bladder calculus before the index date for cases was 3.42 (95% CI = 2.48-4.72) when compared with controls after adjusting for monthly income, geographic region, hypertension, diabetes, coronary heart disease, and renal disease, tobacco use disorder, obesity, alcohol abuse, and schistosomiasis, bladder outlet obstruction, and urinary tract infection. We further analyzed according to sex and found that among males, the OR of having been previously diagnosed with bladder calculus for cases was 3.45 (95% CI = 2.39-4.99) that of controls. Among females, the OR was 3.05 (95% CI = 1.53-6.08) that of controls. These results add to the evidence surrounding the conflicting reports regarding the association between BC and prior bladder calculus and highlight a potential target population for bladder cancer screening.


Therapeutic touch for healing acute wounds.
Therapeutic Touch (TT) as a treatment modality has increased in popularity over the past two decades. Its efficacy in relation to the healing of wounds has been expounded in anecdotal publications. To date however there has been no systematic review. To identify and review all relevant data to determine the effectiveness of TT in the promotion of wound healing. The Cochrane Wounds Group Specialised Trials Register and the Cochrane Central Register of Controlled Trials were searched in May 2003. The Cochrane Complementary Medicine field database was searched in September 2002. In addition the Sigma Theta Tau International Registry of Nursing Research (1983 to 2002) was also searched in September 2002. Furthermore searches of MEDLINE (1966 to 2003) were conducted in 1997 and May 2003, CINAHL (1982 to 2002) in September 2002 and dissertation abstracts in September 2002. All randomised or quasi-randomised controlled trials, which compared the effect of TT with a placebo, another treatment, or no treatment control were considered. Studies which used TT as a stand-alone treatment, or as an adjunct to other therapies, were eligible. One reviewer (DO) determined the eligibility for inclusion of all trials. Both reviewers conducted data extraction and evaluation of each trial's quality independently. Each trial was analysed based on predetermined criteria and a narrative of each trial's main conclusions was produced. Four trials in people with experimental wounds were eligible for inclusion. The effect of TT on wound healing in these studies was variable. Two of the studies (n = 44 & 24) demonstrated a significant effect of TT. However the results of the two other trials favoured the control group (n = 15 & 38), one of these trials demonstrated a significant effect in favour of the control group. There is insufficient evidence that TT promotes healing of acute wounds.


Developmental toxicity of diphenyl ether herbicides in nestling American kestrels.
Beginning the day after hatching, American kestrel (Falco sparverius) nestlings were orally dosed for 10 consecutive days with 5 microliters/g of corn oil (controls) or one of the diphenyl ether herbicides (nitrofen, bifenox, or oxyfluorfen) at concentrations of 10, 50, 250, or 500 mg/kg in corn oil. At 500 mg/kg, nitrofen resulted in complete nestling mortality, bifenox in high (66%) mortality, and oxyfluorfen in no mortality. Nitrofen at 250 mg/kg reduced nestling growth as reflected by decreased body weight, crown-rump length, and bone lengths including humerus, radius-ulna, femur, and tibiotarsus. Bifenox at 250 mg/kg had less effect on growth than nitrofen, but crown-rump, humerus, radius-ulna, and femur were significantly shorter than controls. Liver weight as a percent of body weight increased with 50 and 250 mg/kg nitrofen. Other manifestations of impending hepatotoxicity following nitrofen ingestion included increased hepatic GSH peroxidase activity in all nitrofen-treated groups, and increased plasma enzyme activities for ALT, AST, and LDH-L in the 250-mg/kg group. Bifenox ingestion resulted in increased hepatic GSH peroxidase activity in the 50- and 250-mg/kg groups. Nitrofen exposure also resulted in an increase in total plasma thyroxine (T4) concentration. These findings suggest that altricial nestlings are more sensitive to diphenyl ether herbicides than young or adult birds of precocial species.


Custom impression coping for an exact registration of the healed tissue in the esthetic implant restoration.
It is well known that guided soft tissue healing with a provisional restoration is essential to obtain optimal anterior esthetics in the implant prosthesis. What is not well known is how to transfer a record of beautiful anatomically healed tissue to the laboratory. With the advent of emergence profile healing abutments and corresponding impression copings, there has been a dramatic improvement over the original 4.0-mm diameter design. This is a great improvement, however, it still does not accurately transfer a record of anatomically healed tissue, which is often triangularly shaped, to the laboratory, because the impression coping is a round cylinder. This article explains how to fabricate a "custom impression coping" that is an exact record of anatomically healed tissue for accurate duplication. This technique is significant because it allows an even closer replication of the natural dentition.


Analytical validation of a conventional cardiac troponin I assay for dogs and cats.
Cardiac troponins are gold-standard biomarkers of myocardial injury. There is a need for validation of assays with higher availability and lower costs in veterinary medicine. The primary aim of the present study was to perform an analytical validation of the IMMULITE 2000 TnI assay for use in dogs and cats. A secondary aim was to evaluate its agreement with the previously validated and sensitive Siemens ADVIA Centaur TnI-Ultra assay. Intra- and inter-assay variation, detection limits, the linearity under dilution, and a sample addition study (modified spike-and-recovery analysis) were investigated to assess analytical performance in 15 canine and 15 feline serum samples. Agreement between the assays was evaluated by correlation and Bland-Altman analyses including an additional 99 canine serum samples. Intra-assay variation of cTnI in canine and feline serum was 3.71% and 4.68%, while inter-assay variation was 5.88% and 6.54%, respectively. The assay performed with acceptable linearity within a clinically relevant range of serum cTnI concentrations. The sample addition study revealed insufficient recovery in the range of 71.9%-81.4% for dogs and 62.6%-75.7% for cats. This was considered to be due to a negative matrix effect. A significant correlation between the assays was found, and the Bland-Altman analysis showed acceptable agreement for a wide range of concentrations, but revealed a proportional error, with the IMMULITE TnI assay consistently measuring a higher concentration than the Centaur TnI-Ultra assay. This was relevant only at high serum cTnI concentrations. The IMMULITE TnI assay is considered acceptable for clinical use in dogs and cats.


Long-term outcomes of simultaneous heart and kidney transplantation in pediatric recipients.
Pediatric sHKTx has become an effective therapy for patients with combined cardiac and renal failure. Often, these patients develop human leukocyte antigen antibodies from their previous allografts and are therefore more difficult to re-transplant. We describe the largest case series of a predominantly sensitized pediatric sHKTx with emphasis on medical management and patient outcomes. Demographics, clinical characteristics, antibody, and biopsy data were retrospectively collected from University of California, Los Angeles database and correlated with short- and long-term patient and allograft outcomes of all sHKTx performed between 2002 and 2015. We identified seven pediatric patients who underwent sHKTx at our center. Mean age at time of sHKTx was 13.7 years and 85.7% were re-graft patients. 57.1% were sensitized with cPRA >50% and another 57.1% had preformed donor-specific antibody. Five-year renal allograft survival and patient survival was 85.7% for both end-points. The remaining six patients are all alive (mean follow-up 78.5 months) with good kidney and heart function. sHKTx in a population with increased immunological risk can be associated with good long-term outcomes and offers potential guidance to the pediatric transplant community where data are limited.


Pharmacokinetics and pharmacodynamics of sildenafil in fetal lambs on extracorporeal support.
Maternal transplacental administration of sildenafil is being considered for a variety of fetal conditions. Clinical translation also requires evaluation of fetal safety in a higher species, such as the fetal lamb. Experiments with the pregnant ewe are curtailed by minimal transplacental transfer as well as limited access to the fetus. The EXTra-uterine Environment for Neonatal Development (EXTEND) model renders the isolated fetal lamb readily accessible and allows for direct fetal administration of sildenafil. Five fetal lambs were placed on extracorporeal support in the EXTEND device and received continuous intravenous (IV) sildenafil (0.3-0.5-0.7 mg/kg/24hr) for a duration of one to seven days. Plasma sildenafil concentrations were sampled at regular intervals to establish the pharmacokinetic profile using population pharmacokinetic modeling. Serial Doppler ultrasound examination, continuous non-invasive hemodynamic monitoring and blood gas analysis were done to evaluate the pharmacodynamic effects and fetal response. The target concentration range (47-500 ng/mL) was attained with all doses. Sildenafil induced an immediate and temporary reduction of pulmonary vascular resistance, mean arterial pressure and circuit flow, without change in fetal lactate levels and acid-base status. The duration of the systemic effects increased with the dose. Immediate temporary pulmonary vascular and systemic hemodynamic changes induced by sildenafil were biochemically well tolerated by fetal lambs on extracorporeal support, with the 0.5 mg/kg/24 h dose balancing rapid attainment of target concentrations with short-lived systemic effects. None. A literature review was conducted searching online databases (Medline, Embase and Cochrane), using search terms: fetal OR prenatal OR antenatal AND sildenafil, without time-limit and excluding human studies. Where relevant, investigators were contacted in order to avoid duplication of work. Prenatal therapy with sildenafil, a phosphodiesterase-5 inhibitor with vasodilatory and anti-remodeling effects on vascular smooth muscle cells, has been considered for a variety of fetal conditions. One multicenter clinical trial investigating the benefit of sildenafil in severe intrauterine growth restriction (the STRIDER-trial) was halted early due to excess mortality in the sildenafil-exposed arm at one treatment site. Such findings demonstrate the importance of extensive preclinical safety assessment in relevant animal models. Transplacentally administered sildenafil leads to decreased pulmonary arterial muscularization, preventing or reducing the occurrence of pulmonary hypertension in rat and rabbit fetuses with diaphragmatic hernia (DH). Validation of these results in a higher and relevant animal model, e.g. fetal lambs, is the next step to advance clinical translation. We recently demonstrated that, in contrast to humans, transplacental transfer of sildenafil in sheep is minimal, precluding the in vivo study of fetal effects at target concentrations using the conventional pregnant ewe model. We therefore used the extracorporeal support model for fetal lambs, referred to as the EXTra-uterine Environment for Neonatal Development (EXTEND) system, bypassing placental and maternal metabolism, to investigate at what dose the target concentrations are reached, and what the fetal hemodynamic impact and response are. Fetal hemodynamic and metabolic tolerance to sildenafil are a crucial missing element on the road to clinical translation. This is therefore the first study investigating the pharmacokinetics, hemodynamic and biochemical effects of clinical-range concentrations of sildenafil in fetal lambs, free from placental and maternal interference. We demonstrated self-limiting pulmonary vasodilation, a decrease of both systemic arterial pressures and circuit flows, induced by clinical range concentrations of sildenafil, without the development of fetal acidosis. This paves the way for further investigation of prenatal sildenafil in fetal lambs on extracorporeal support. A dose of 0.5 mg/kg/24 h offered the best trade-off between rapid achievement of target concentrations and shortest duration of systemic effects. This is also the first study using the EXTEND as a model for pharmacotherapy during pregnancy.


The UBE2C/CDH1/DEPTOR axis is an oncogene and tumor suppressor cascade in lung cancer cells.
Ubiquitin-conjugating enzyme E2C (UBE2C) mediates ubiquitylation chain formation via the K11 linkage. While previous in vitro studies showed that UBE2C plays a growth-promoting role in cancer cell lines, the underlying mechanism remains elusive. Still unknown is whether and how UBE2C plays a promoting role in vivo. Here we report that UBE2C was indeed essential for growth and survival of lung cancer cells harboring Kras mutations, and UBE2C was required for KrasG12D-induced lung tumorigenesis, since Ube2c deletion significantly inhibited tumor formation and extended the lifespan of mice. Mechanistically, KrasG12D induced expression of UBE2C, which coupled with APC/CCDH1 E3 ligase to promote ubiquitylation and degradation of DEPTOR, leading to activation of mTORC signaling. Importantly, DEPTOR levels fluctuated during cell cycle progression in a manner dependent on UBE2C and CDH1, indicating their physiological connection. Finally, Deptor deletion fully rescued the tumor inhibitory effect of Ube2c deletion in the KrasG12D lung tumor model, indicating a causal role of Deptor. Taken together, our study shows that the UBE2C/CDH1/DEPTOR axis forms an oncogene and tumor suppressor cascade that regulates cell cycle progression and autophagy and validates UBE2C an attractive target for lung cancer associated with Kras mutations.


Successful breeding predicts divorce in plovers.
When individuals breed more than once, parents are faced with the choice of whether to re-mate with their old partner or divorce and select a new mate. Evolutionary theory predicts that, following successful reproduction with a given partner, that partner should be retained for future reproduction. However, recent work in a polygamous bird, has instead indicated that successful parents divorced more often than failed breeders (Halimubieke et al. in Ecol Evol 9:10734-10745, 2019), because one parent can benefit by mating with a new partner and reproducing shortly after divorce. Here we investigate whether successful breeding predicts divorce using data from 14 well-monitored populations of plovers (Charadrius spp.). We show that successful nesting leads to divorce, whereas nest failure leads to retention of the mate for follow-up breeding. Plovers that divorced their partners and simultaneously deserted their broods produced more offspring within a season than parents that retained their mate. Our work provides a counterpoint to theoretical expectations that divorce is triggered by low reproductive success, and supports adaptive explanations of divorce as a strategy to improve individual reproductive success. In addition, we show that temperature may modulate these costs and benefits, and contribute to dynamic variation in patterns of divorce across plover breeding systems.


High-intensity ultrasound promoted the maturation of high-salt liquid-state soy sauce: A mean of enhancing quality attributes and sensory properties.
High-intensity ultrasound was used as a means to promote maturation of soy sauce. The optimal conditions for ultrasound treatment were 90℃ at an ultrasound intensity of 39.48 W/cm2 for 60 min. The total reducing sugars and soluble salt-free solids content was significantly increased after ultrasound-assisted maturation. The free amino acid content was significantly decreased, mainly due to the Maillard reaction (MR). The promoted MR produced several types of flavor compounds, including esters, pyrazines, and ketones, which imparted an attractive aroma to the maturated soy sauce. The proportion of peptides with a molecular weight of 1-5 kDa provided umami as an important flavor characteristic, and the content in the ultrasound-matured soy sauce (10.19 %) was significantly higher than that in the freshly prepared soy sauce (8.34 %) and the thermally treated sample (8.89 %). Ultrasound-assisted maturation would improve product quality and meanwhile, shorten the duration and reduce the cost for the soy sauce industry.


Quantitative analysis of acetates in cigarette tobacco using solid-phase microextraction and gas chromatography-mass spectrometry.
A method incorporating solid-phase microextraction (SPME) and gas chromatography-mass spectrometry for the headspace analysis of selected volatile organic compounds present in cigarette tobacco is developed and evaluated. Quantitative information on methyl, ethyl, n-propyl, isopropyl, isopropenyl, vinyl, and butyl acetates present in 29 different flavor variants (full, light, and ultra-light) of the top ten selling brands in the United States is presented. The concentrations of the various acetate analytes range from the low nanaogram to microgram levels per cigarette. Clear differences are observed in the concentrations of various acetates when comparing the levels in brands from different manufacturers. The SPME technique provides a method that allows high sample throughput, requires little sample preparation, and yields useful analytical information. High precision is obtained on multiple measurements of cigarettes from an individual pack, but lower precision levels are observed in general when comparing results obtained on the analysis of cigarettes from different packs of the same brand. The higher pack-to-pack variations may be due in part to product aging with a proportionate amount of evaporative loss of the relatively volatile acetates.


Adult-to-adult living-donor liver transplantation: a critical analysis.
Adult-to-adult living-donor liver transplantation is the most recent achievement in the evolution of strategies to increase donor organ supply. Justification of the procedure has evolved from increased organ-waiting times, wait-list morbidity and mortality of adult transplant candidates. Successful application of adult-to-adult living-donor liver transplantation mandates unique surgical, medical, and donor considerations as these procedures predispose to unique complications resulting from anatomic variations, technical considerations, and the transplantation of "partial" grafts. Additionally, because the magnitude of the donor operation is greater and potentially fraught with serious short and long term morbidity and possibly mortality, the utilization of living-donors for adult liver transplantation raises bio-ethical concerns heretofore not encountered. This review explores the current state of adult-to-adult living-donor liver transplantation.


A new social communication intervention for children with autism: pilot randomised controlled treatment study suggesting effectiveness.
Psychosocial treatments are the mainstay of management of autism in the UK but there is a notable lack of a systematic evidence base for their effectiveness. Randomised controlled trial (RCT) studies in this area have been rare but are essential because of the developmental heterogeneity of the disorder. We aimed to test a new theoretically based social communication intervention targeting parental communication in a randomised design against routine care alone. The intervention was given in addition to existing care and involved regular monthly therapist contact for 6 months with a further 6 months of 2-monthly consolidation sessions. It aimed to educate parents and train them in adapted communication tailored to their child's individual competencies. Twenty-eight children with autism were randomised between this treatment and routine care alone, stratified for age and baseline severity. Outcome was measured at 12 months from commencement of intervention, using standardised instruments. All cases studied met full Autism Diagnostic Interview (ADI) criteria for classical autism. Treatment and controls had similar routine care during the study period and there were no study dropouts after treatment had started. The active treatment group showed significant improvement compared with controls on the primary outcome measure--Autism Diagnostic Observation Schedule (ADOS) total score, particularly in reciprocal social interaction--and on secondary measures of expressive language, communicative initiation and parent-child interaction. Suggestive but non-significant results were found in Vineland Adaptive Behaviour Scales (Communication Sub-domain) and ADOS stereotyped and restricted behaviour domain. A Randomised Treatment Trial design of this kind in classical autism is feasible and acceptable to patients. This pilot study suggests significant additional treatment benefits following a targeted (but relatively non-intensive) dyadic social communication treatment, when compared with routine care. The study needs replication on larger and independent samples. It should encourage further RCT designs in this area.


The Great American Biotic Interchange in frogs: multiple and early colonization of Central America by the South American genus Pristimantis (Anura: Craugastoridae).
The completion of the land bridge between North and South America approximately 3.5-3.1 million years ago (Ma) initiated a tremendous biogeographic event called the Great American Biotic Interchange (GABI), described principally from the mammalian fossil record. The history of biotic interchange between continents for taxonomic groups with poor fossil records, however, is not well understood. Molecular and fossil data suggest that a number of plant and animal lineages crossed the Isthmus of Panama well before 3.5 Ma, leading biologists to speculate about trans-oceanic dispersal mechanisms. Here we present a molecular phylogenetic analysis of the frog genus Pristimantis based on 189 individuals of 137 species, including 71 individuals of 31 species from Panama and Colombia. DNA sequence data were obtained from three mitochondrial (COI, 12S, 16S) and two nuclear (RAG-1 and Tyr) genes, for a total of 4074 base pairs. The resulting phylogenetic hypothesis showed statistically significant conflict with most recognized taxonomic groups within Pristimantis, supporting only the rubicundus Species Series, and the Pristimantis myersi and Pristimantis pardalis Species Groups as monophyletic. Inference of ancestral areas based on a likelihood model of geographic range evolution via dispersal, local extinction, and cladogenesis (DEC) suggested that the colonization of Central America by South American Pristimantis involved at least 11 independent events. Relaxed-clock analyses of divergence times suggested that at least eight of these invasions into Central America took place prior to 4 Ma, mainly in the Miocene. These findings contribute to a growing list of molecular-based biogeographic studies presenting apparent temporal conflicts with the traditional GABI model.


Fouling assemblage of benthic plastic debris collected from Mersin Bay, NE Levantine coast of Turkey.
The Mediterranean is an ecosystem that faces more and more microplastic pollution every day. This causes the whole of the Mediterranean to face the negative effects of plastic pollution. This study examines the state of plastic debris and fouling organisms found on it in one of the areas most affected by plastic pollution, Mersin Bay. As a result, a total of 3.88kg plastic (mean=0,97kg; n=120; 2670item/km2; 86,3kg/km2) was collected and based on the ATR-FTIR analysis, it was determined that this total contained 9 types of plastics. 17 different fouling species belonging to 6 phylum (Annelida, Arthropoda, Bryozoa, Chordata, Cnidaria, Mollusca) 7 class and 11 order were discovered on plastics. Spirobranchus triqueter, Hydroides sp. and Neopycnodonte cochlear were the most abundant species. In the end, the example of Mersin Bay shows that plastic debris as a substrate can contain a very high diversity of life just like natural substrates.


Chevron osteotomy and single wire reattachment of the greater trochanter in primary and revision total hip arthroplasty.
Even at a medical center where hip arthroplasty is performed routinely without trochanteric osteotomy, selected arthroplasties still require greater trochanteric osteotomy. A technique of greater trochanteric biplane osteotomy and reattachment with one wire is described. The technique provides maximum stability with a minimum of internal fixation and it can be performed quickly. The postoperative rehabilitation protocol is the same as that used in hip arthroplasty patients without trochanteric osteotomy. Results with a minimum two-year follow-up period disclosed osseous union in 98% of 53 primary arthroplasties and 97% of 74 revision hip arthroplasties. Problems related to the greater trochanter necessitated reoperation in only two patients: one for reattachment of a migrated trochanter and one for trochanteric wire removal.


GMI ablates cancer stemness and cisplatin resistance in oral carcinomas stem cells through IL-6/Stat3 signaling inhibition.
Cancer stem cells (CSCs) have been identified to exert tumor-initiating ability, resulting in the recurrence, metastasis and chemoresistance of oral squamous cell carcinomas. In the present study, we showed that GMI, an immunomodulatory protein from Ganoderma microsporum, induc ed a cytotoxic effect in oral carcinomas stem cells (OCSCs). Treatment of GMI dose-dependently inhibited the expression of CSC markers, including ALDH1 activity and CD44 positivity. Moreover, GMI suppressed the self-renewal property, colony formation, migration, and invasion abilities as well as potentiated chemo-sensitivity in OCSCs. Our results suggested that the tumor suppressive effect of GMI was mediated through inhibition of IL-6/Stat3 signaling pathway. Furthermore, tumor growth was reduced in mice bearing xenograft tumors after oral administration of GMI. Taken together, we demonstrated the anti-CSC effect of GMI in oral cancer and GMI may serve as a natural cisplatin adjuvant to prevent cancer recurrence.


Simple construction of Cu2-xS:Pt nanoparticles as nanotheranostic agent for imaging-guided chemo-photothermal synergistic therapy of cancer.
Synergistic therapy has attracted intense attention in medical treatment because it can make up for the disadvantages of single therapy and greatly improve the efficacy of cancer treatment. However, it remains a challenge to build a simple system to achieve synergistic therapy. In this study, X-ray computed tomography (CT) imaging-guided chemo-photothermal synergistic therapy can be easily achieved by simple construction of Cu2-xS:Pt(0.3)/PVP nanoparticles (NPs). Cu2-xS:Pt(0.3)/PVP NPs can passively accumulate within the tumor sites, thus ensuring that many Cu2-xS:Pt(0.3)/PVP NPs are brought into the tumor cells, which can be confirmed by the results of cellular uptake, imaging, and nanoparticle biodistribution. It can be verified that the platinum ions can be released from Cu2-xS:Pt(0.3)/PVP NPs under 808 nm laser irradiation. Simultaneously, Pt(iv) ions are reduced to Pt(ii) ions by excess glutathione and then, they exhibit chemo-anticancer activities. In addition, Cu2-xS:Pt(0.3)/PVP NPs can be used as an effective photothermal agent. The results demonstrate that the efficient tumor growth inhibition effect can be realized from the mice treated with Cu2-xS:Pt(0.3)/PVP NPs under 808 nm laser irradiation by chemo-photothermal synergistic therapy. Furthermore, Cu2-xS:Pt(0.3)/PVP NPs can be thoroughly cleared through feces in a short time, showing high biosafety for further potential clinical translations.


Structures of the P. aeruginosa FleQ-FleN master regulators reveal large-scale conformational switching in motility and biofilm control.
Pseudomonas aeruginosa can cause a wide array of chronic and acute infections associated with its ability to rapidly switch between planktonic, biofilm, and dispersed lifestyles, each with a specific arsenal for bacterial survival and virulence. At the cellular level, many of the physiological transitions are orchestrated by the intracellular second messenger c-di-GMP and its receptor-effector FleQ. A bacterial enhancer binding protein, FleQ acts as a master regulator of both flagellar motility and adherence factor secretion and uses remarkably different transcription activation mechanisms depending on its dinucleotide loading state, adenosine triphosphatase (ATPase) activity, interactions with polymerase sigma (σ) factors, and complexation with a second ATPase, FleN. How the FleQ-FleN tandem can exert diverse effects through recognition of a conserved FleQ binding consensus has remained enigmatic. Here, we provide cryogenic electron microscopy (cryo-EM) structures of both c-di-GMP-bound and c-di-GMP-free FleQ-FleN complexes which deepen our understanding of the proteins' (di)nucleotide-dependent conformational switching and fine-tuned roles in gene expression regulation.


The current role of invasive staging in lung cancer.
Precise mediastinal lymph node staging in otherwise operable non-small cell lung cancer (NSCLC) is imperative as it determines subsequent treatment and prognosis. For mediastinal staging, even present-day computed tomography (CT) scanners have a low accuracy, and routine cervical mediastinoscopy is advised for precise lymph node assessment, certainly when considering a neoadjuvant protocol. Cervical mediastinoscopy remains the gold standard, and sampling of subcarinal nodes is essential. The role of remediastinoscopy after induction chemo- or radiotherapy remains to be determined. Extended mediastinoscopy is indicated to take biopsies of scalene lymph nodes or aorto-pulmonary nodes, which can also be reached by anterior mediastinoscopy. Video-assisted thoracic surgery (VATS) does not replace cervical mediastinoscopy but is a valid alternative to anterior or extended mediastinoscopy. A more complete exploration of the ipsilateral hemithorax is possible, with biopsies of aortopulmonary and inferior mediastinal nodes and judgment about resectability of the primary tumour.


Epidemiologic impact of invasion and post-invasion conflict in Iraq.
There has been little systematic analysis of casualty data from the Iraq invasion and postinvasion conflict since 2003. Here we combine well known sources on military casualties and little known or understood sources on non-combatant mortality to identify major trends and impacts. This conflict is unique in many ways. It is associated with high risk of death to previously little affected groups ? female and older adult soldiers. From the early days of combat, the conflict has resulted in a relatively high rate of death among soldiers, reversing a long term trend toward declining mortality among U.S. troops. Despite a high survival rate among those with serious injuries, it is the first conflict for which most deaths occurred after the end of major hostilities. Deaths among non-combatant groups are much higher, and have resulted in far more confusion regarding actual mortality rates. This is not surprising; in few of the major conflicts or humanitarian crisis have epidemiologic estimates been made. It is shown that pre-invasion projections regarding civilian casualties were uniformly mistaken regarding the major risks and risk levels to be faced. More research is needed to improve and standardize approaches to identifying mortality risk to major population groups.