Cloning, overexpression, crystallization and preliminary X-ray crystallographic analysis of a slow-processing mutant of penicillin G acylase from Kluyvera citrophila.
Kluyvera citrophila penicillin G acylase (KcPGA) has recently attracted increased attention relative to the well studied and commonly used Escherichia coli PGA (EcPGA) because KcPGA is more resilient to harsh conditions and is easier to immobilize for the industrial hydrolysis of natural penicillins to generate the 6-aminopenicillin (6-APA) nucleus, which is the starting material for semi-synthetic antibiotic production. Like other penicillin acylases, KcPGA is synthesized as a single-chain inactive pro-PGA, which upon autocatalytic processing becomes an active heterodimer of α and β chains. Here, the cloning of the pac gene encoding KcPGA and the preparation of a slow-processing mutant precursor are reported. The purification, crystallization and preliminary X-ray analysis of crystals of this precursor protein are described. The protein crystallized in two different space groups, P1, with unit-cell parameters a = 54.0, b = 124.6, c = 135.1 Å, α = 104.1, β = 101.4, γ = 96.5°, and C2, with unit-cell parameters a = 265.1, b = 54.0, c = 249.2 Å, β = 104.4°, using the sitting-drop vapour-diffusion method. Diffraction data were collected at 100 K and the phases were determined using the molecular-replacement method. The initial maps revealed electron density for the spacer peptide.


Overexpression of the ribosomal S30 subunit leads to indole-3-carbinol tolerance in Arabidopsis thaliana.
Indole-3-carbinol (I3C), a hydrolysis product of indole-3-methylglucosinolate, is toxic to herbivorous insects and pathogens. In mammals, I3C is extensively studied for its properties in cancer prevention and treatment. Produced in Brassicaceae, I3C reversibly inhibits root elongation in a concentration-dependent manner. This inhibition is partially explained by the antagonistic action of I3C on auxin signaling through TIR1. To further elucidate the mode of action of I3C in plants, we have identified and characterized a novel Arabidopsis mutant tolerant to I3C, ICT1. This mutant was identified following screening of the Full-length cDNA Over-eXpression library (FOX) seed collection for root growth in the presence of exogenous I3C. ICT1 carries the AT2G19750 gene, which encodes an S30 ribosomal protein. Overexpression, but not knockout, of the S30 gene causes tolerance to I3C. The tolerance is specific to I3C, since ICT1 did not exhibit pronounced tolerance to other indole or benzoxazinoid molecules tested. ICT1 maintains I3C-induced antagonism of auxin signaling, indicating that the tolerance is due to an auxin-independent mechanism. Transcript profiling experiments revealed that ICT1 is transcriptionally primed to respond to I3C treatment.


Evolution of postural stability after subthalamic nucleus stimulation in Parkinson's disease: a combined clinical and posturometric study.
The occurrence of postural and balance disorders is a frequent feature in advanced forms of Parkinson's disease (PD). However, the pathological substrate of these disturbances is poorly understood. In the present work, we investigated the evolution of posturometric parameters [center of pressure (CoP) displacement and CoP area] and axial scores between the pre-operative period and 3 months post-operative in seven PD patients who underwent bilateral deep brain stimulation (DBS) of the subthalamic nucleus (STN). After surgery, the patients leaned backwards much more regardless of the STN stimulation, suggesting that surgery could have a deleterious effect on postural adaptation. During the post-operative period, the improvement in axial and postural scores was similar under levodopatherapy and DBS. On the other hand, DBS of the STN significantly reduced the CoP displacement and the CoP area, whereas levodopatherapy tended only to reduce the CoP displacement and to increase the CoP area significantly. These data suggest that DBS of the STN and levodopa do not act on the same neurological systems involved in posture regulation. DBS of the STN could improve posture via a direct effect on the pedunculopontine nucleus, which is known to be involved in posture regulation.


Characterization of ram (Ovis aries) sperm head morphometry using the Sperm-Class Analyzer.
Sperm morphology has been identified as a characteristic that can be useful in the prediction of fertilizing capacity. The aim of the current study was to characterize ram sperm heads morphometrically as a basis for future studies on the relationship between sperm quality and male fertility. For this purpose, ejaculates from 241 mature rams (Ovis aries) belonging to 36 different dairy herds were used to evaluate sperm head morphometry by means of the Sperm-Class Analyzer. Sperm samples, collected by artificial vagina, were diluted in phosphate buffered saline (PBS) for the analysis. A microscope slide was prepared from single-diluted fresh sperm samples. Slides were air-dried and stained with Hemacolor. A minimum of 115 sperm heads were analyzed from each male. Each sperm head was measured for four primary parameters (area, perimeter, length, width), and four derived parameters of head shape were obtained. Significant differences in sperm head morphometry were found between rams (CV for morphometric parameters ranging from 0.9 to 10.1), and there were marked differences in the sperm morphometric composition of the ejaculates. For all parameters, within-animal CVs were greater than between-animal CVs. Within-animal CVs ranged from 4.2 to 10.6, showing the high degree of sperm polymorphism present in the sheep ejaculate. Significant differences in sperm head morphometry were found between rams belonging to the different herds (i.e., origin). An important part of the variability observed on morphometric parameters was due to the male itself, with an explained variance ranging from 3.6% for regularity to 34.0% for p2a (perimeter(2)/[4xpixarea]). The explained variance by the herd of origin of the males ranged from 0.6% for regularity to 10.8% for area. Our results suggest that a genetic component might be responsible for the observed sperm head morphometry differences between herds.


Mechanical stretch and hypoxia inducible factor-1 alpha affect the vascular endothelial growth factor and the connective tissue growth factor in cultured ACL fibroblasts.
The adult human anterior cruciate ligament (ACL) has poor functional healing response. Hypoxia plays an important role in regulating the microenvironment of the joint cavity after ACL injury, however, its role in mechanical injury is yet to be examined fully in ACL fibroblasts. In this study, we used CoCl2 to induce Hypoxia-inducible factor-1α (HIF-1α) in our experimental model to study its affect on matrix metalloproteinase-2 (MMP-2), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) expression in ACL fibroblasts after mechanical stretch. Cell treatments were performed in the stretch chamber in all experimental groups. Quantitative real-time PCR was used to check mRNA expression levels of MMP-2, CTGF, VEGF, and HIF-1α. Western blot was used to detect the HIF-1α production. Enzyme-Linked immunosorbent assay was performed to check the VEGF and CTGF protein contents in supernatant. MMP-2 activity was assayed by gelatin zymography. The real-time PCR results show that mechanical stretch or CoCl2 treatment increases the expression of MMP-2, VEGF, CTGF, and HIF-1α; however, the combined effects of mechanical stretch and CoCl2-induced HIF-1α increased MMP-2 production but decreased the VEGF and CTGF expression, compared to the CoCl2 treatment group alone. Western blot analysis and ELISA also confirmed these results. Our results demonstrated that mechanical stretch and CoCl2-induced HIF-1α together increased the level of MMP-2 and decreased the levels of VEGF and CTGF in cultured ACL fibroblasts. The differential expression and production of HIF-1α, VEGF, MMP-2, and CTGF might help to explain the poor healing ability of ACL.


Associations between respiratory signs, thoracic CT findings and results of tracheobronchoscopy and bronchoalveolar lavage in dogs.
Several diagnostic techniques are used in dogs with signs of respiratory disease. The aims of the present study are to estimate the relative sensitivities and associations between the results of diagnostic tests in dogs with respiratory conditions. A retrospective cross-sectional study of dogs referred for investigation of respiratory signs. Associations between clinical signs, thoracic CT findings, tracheobronchoscopic findings, cytology results and bacterial culture results were tested using binary logistic regression. One hundred and thirty-three dogs were included. Abnormalities were detected by cytology, tracheobronchoscopy, CT and bacterial culture in 91%, 88%, 80% and 25% cases, respectively. There were associations between cough and bronchial lesions on thoracic CT (odds ratio [OR] 2.6, 95% confidence interval [CI] 1.1-6.4, p = 0.037), and between cough and neutrophilic inflammation on cytology (OR 4.5, 95% CI 1.3-15.8, p = 0.020). Bronchial foreign body at bronchoscopy was associated with pulmonary consolidation on CT (OR 8.0, 95% CI 1.6-41.7, p = 0.013) and with positive bacterial culture (OR 10.9, 95% CI 2.1-57.0, p = 0.005). In dogs with normal thoracic CT, abnormalities were detected by cytology, tracheobronchoscopy and bacterial culture in 89%, 77% and 23% cases, respectively. Airway cytology and tracheobronchoscopy provided useful information for diagnosis in many dogs with respiratory signs that had a normal thoracic CT.


Fatigue and herpesvirus latency in women newly diagnosed with breast cancer.
Fatigue is a notable clinical problem in cancer survivors, and understanding its pathophysiology is important. The current study sought to determine biomarkers of fatigue that exist before cancer treatment. Relationships between the expression of latent Epstein-Barr virus (EBV) and cytomegalovirus (CMV) and fatigue were examined in 158 women newly diagnosed with breast cancer or awaiting a positive diagnostic result. Higher CMV antibody titers, but not EBV antibody titers, were associated with a greater likelihood of being fatigued. Associations between fatigue and higher CMV antibody titers remained after controlling for alcohol use, smoking, comorbidities, depressive symptoms, age, BMI, cancer stage, and sleep problems. More sleep problems and higher levels of depressive symptoms were also associated with a greater likelihood of being fatigued. CMV antibody titers, but not EBV antibody titers, were associated with higher levels of C-reactive protein (CRP), but CRP was not associated with fatigue. When the cellular immune system is compromised, reactivation of latent herpesviruses may fuel chronic inflammatory responses. Prior work has suggested that fatigue may be related to inflammation and its associated sickness behaviors; accordingly, our findings may be tapping into this same physiological substrate.


Bmp2 is essential for cardiac cushion epithelial-mesenchymal transition and myocardial patterning.
Cardiac cushion development provides a valuable system to investigate epithelial to mesenchymal transition (EMT), a fundamental process in development and tumor progression. In the atrioventricular (AV) canal, endocardial cells lining the heart respond to a myocardial-derived signal, undergo EMT, and contribute to cushion mesenchyme. Here, we inactivated bone morphogenetic protein 2 (Bmp2) in the AV myocardium of mice. We show that Bmp2 has three functions in the AV canal: to enhance formation of the cardiac jelly, to induce endocardial EMT and to pattern the AV myocardium. Bmp2 is required for myocardial expression of Has2, a crucial component of the cardiac jelly matrix. During EMT, Bmp2 promotes expression of the basic helix-loop-helix factor Twist1, previously implicated in EMT in cancer metastases, and the homeobox genes Msx1 and Msx2. Deletion of the Bmp type 1A receptor, Bmpr1a, in endocardium also resulted in failed cushion formation, indicating that Bmp2 signals directly to cushion-forming endocardium to induce EMT. Lastly, we show that Bmp2 mutants failed to specify the AV myocardium with loss of Tbx2 expression uncovering a myocardial, planar signaling function for Bmp2. Our data indicate that Bmp2 has a crucial role in coordinating multiple aspects of AV canal morphogenesis.


Blending Donors with Different Molecular Weights: An Efficient Strategy to Resolve the Conflict between Coherence Length and Intermixed Phase in Polymer/Nonfullerene Solar Cells.
Long coherence lengths (CLs) of crystals and proper intermixed phase amount guarantee charge transport and exciton dissociate efficiently, which is crucial for organic solar cells (OSCs) to achieve high device performance. However, extending CLs usually reduces the intermixed phase, leading to an insufficient interface for exciton dissociation. Herein, a strategy using a binary polymer with different molecular weights as donor is employed, that is, poly(3-hexylthiophene-2,5-diyl) (P3HT) with high (P3HT-H) and low (P3HT-L) molecular weight are blended as donor, and (5Z,5'Z)-5,5'-(((4,4,9,9-tetraoctyl-4,9-dihydro-s-indaceno[1,2-b:5,6-b']dithiophene-2,7-diyl)bis(benzo[c][1,2,5]thiadiazole-7,4-diyl))bis(methanylylidene))bis(3-ethyl-2-thioxothiazolidin-4-one) (O-IDTBR) is used as acceptor. In kinetics, the entanglements of P3HT-H are relieved due to the higher molecular diffusivity of P3HT-L. In thermodynamics, the miscibility of P3HT-L/O-IDTBR, P3HT-H/O-IDTBR, and P3HT-L/P3HT-H blends increases in turn. Hence, P3HT forms a more ordered structure with longer CLs after adding P3HT-L, which also drives O-IDTBR dispersed in P3HT crystalline regions diffuse to the O-IDTBR crystalline regions to further self-organize. Consequently, the CLs of both P3HT and O-IDTBR are extended, while keeping the intermixed phase amount proper. The optimized microstructure boosts device performance from 7.03% to 7.80%, which is one of the highest values reported for P3HT/O-IDTBR blends. This is a novel way to solve the conflict mentioned above, which may provide guidance to finely regulating the morphology of the active layer.


Transmastoid repair of superior semicircular canal dehiscence.
Objective/Hypothesis Superior semicircular canal (Sup SC) dehiscence syndrome is a rare condition, causing a variety of auditory and vestibular symptoms. The traditional surgical management is a middle cranial fossa, extradural approach to resurface the Sup SC. Recently, a transmastoid approach for plugging of the Sup SC has been developed. We present further data supporting the use of the transmastoid approach in preference to the middle fossa approach. Design This is a retrospective multi-institutional case series. Method We included 10 patients in this case series from two tertiary otology institutions. Sup SC dehiscence was confirmed by correlation of clinical symptoms with positive audiometric, vestibular evoked myogenic potential, and computed tomography findings. A transmastoid approach was used for plugging of the Sup SC. Either a single fenestration was created at the site of dehiscence or separate fenestrations sited ampullopetal and ampullofugal to the dehiscence. Results All patients who underwent this procedure had good symptom control and hearing preservation postoperatively. Conclusion In patients with adequate temporal bone pneumatization, the transmastoid approach provides a safe and effective alternative to the middle cranial fossa approach. This series has demonstrated excellent symptom control and preservation of hearing with the transmastoid approach.


Control of Cajal body number is mediated by the coilin C-terminus.
Cajal bodies (CBs) are nuclear suborganelles implicated in the post-transcriptional maturation of small nuclear and small nucleolar RNAs. The number of CBs displayed by various cell lines and tissues varies, and factors that control CB numbers within a given cell have yet to be described. In this report, we show that specific regions within the C-terminus of coilin, the CB marker protein, are responsible for regulating the number of nuclear foci. Despite the fact that the coilin N-terminal domain is responsible for its self-oligomerization activity, truncation or mutation of predicted sites of phosphorylation in the conserved C-terminal region leads to a striking alteration in the number of nuclear bodies. Similarly, coilin constructs from various species display differential propensities to form nuclear foci when expressed in heterologous backgrounds. We mapped the domain responsible for this variability to the coilin C-terminus utilizing chimeric proteins. Furthermore, the activities responsible for regulating coilin self-association must reside in the nucleus, as constructs lacking critical nuclear localization sequences fail to form foci in the cytoplasm. Factors controlling the putative signal transduction cascade that phosphorylates coilin are also discussed. The results point to a model whereby phosphorylation of the coilin C-terminus regulates the availability of the N-terminal self-interaction domain.


First-phase insulin secretion is positively correlated with alanine aminotransferase in young adults.
Type 2 diabetes (T2D) is known to be one of the most prevalent diseases, and its prevalence is significantly associated with age and metabolic syndrome (MetS). Few studies have been conducted on liver function, MetS and insulin secretion among young adults. In the present study, we explored the relationship between the liver function enzyme - alanine aminotransferase (ALT) - and first-phase insulin secretion (FPIS) among young adults. There were 22,971 men and 28,740 women, aged 18-27 years, assigned to subgroups according to the presence of MetS and quartiles of ALT values. Simple correlation was applied to evaluate their relationship. The difference between the slopes of these relationships and FPIS were statistically analyzed with Chris's calculator. Most values for metabolic parameters, including ALT and FPIS, were determined to be relatively high in individuals with MetS. By contrast, individuals with MetS had lower high-density-lipoprotein cholesterol (HDL-C) counts and FPIS. Similar results were observed in the quartiles of ALT. Significant positive results were also found in the linear model. Depending on the ALT level, the slope change of FPIS still demonstrated a positive correlation between ALT and FPIS. This correlation was stronger for men than for women. A positive correlation between ALT and FPIS exists among young adults. Moreover, this correlation was stronger for men than for women. Both the cause and the effect require further investigation.


New species in the genus Thagria Melichar from the Oriental and Australian regions, with a revised key to genera and species and a synoptic catalogue of the genus (Hemiptera: Cicadellidae: Coelidiinae).
The following 21 new species of Thagria from the Oriental and Australian regions are described, illustrated and photographed: T. aenigmatis, sp. nov., T. asperitas sp. nov., T. biretrorsa sp. nov., T. colorata, sp. nov., T. coniunctionis, sp. nov., T. constanti, sp.nov., T. fidelitas sp. nov., T. freytagi sp. nov., T. intorta sp. nov., T. iuxta, sp. nov., T. lobata sp. nov., T. longicatilla sp. nov., T. malenovskyi sp. nov., T. oldfieldi, sp. nov., T. paraunca, sp. nov., T. quadrimaculata, sp. nov., T. quadrispinosa sp. nov., T. trimaculata sp. nov., T. unibasispinosa, sp. nov.; T. unica sp. nov. and T. viraktamathi, sp. nov. Thagria bidentata Xu & Kuoh 1998, preoccupied by Thagria bidentata Nielson 1982 is renamed herein Thagria xui nom. nov. Thagria multicalcara Nielson is suppressed junior synonym of Coelidia inscripta Walker. Thagria hongdoensis Kwon & Lee is suppressed junior synonym of resurrected Coelidia satsumensis Matsumura. Cambodia is a new record for T. longistyla Freytag. Laos is a new record for T. acrodens Freytag, T. boulardi Nielson, T.emeiensis Zhang, T. fuscoscuta Zhang, T. janssoni Nielson, T. obrienae Nielson and T. ungulata Nielson. Vietnam is a new record for T. grandis Nielson and T. marissae Nielson. Intra-inter specific variation involving 6 closely related species (melichari species complex) in Southeast Asia is discussed. Morphology and taxonomic value of the dorsal connective, revised key to species and genera in Thagriini and an updated synoptic catalogue of the genus are also presented. Problematical species in the genus Thagria are reviewed. Distribution of 235 known species and the relationship between clypellus configuration and geographical origin are given.


Comparison of the toxin binding sites of the nicotinic acetylcholine receptor from Drosophila to human.
Recombinant toxin binding proteins have been previously found to provide a convenient experimental system for the study of receptor-ligand recognition (Aronheim et al., 1988). Here, this system has been used to produce the binding sites of the cholinergic receptor derived from seven organisms, Torpedo californica, Xenopus, chick, mouse, calf, human, and Drosophila. These have been compared with respect to their toxin binding capacity. Scatchard analyses show that the KD values of alpha-bungarotoxin binding to the above sites are 63, 536, 150, 3200, 6200, 6470, and 1700 nM, respectively. These results reiterate the importance of alpha 183-204 as a ligand binding site. In order to increase the repertoire of sites available for study, chimeric structures were constructed. Through the analysis of such chimeras, some themes of the gross anatomy of the binding site can be learned. A positive subsite followed by a hydrophobic patch preceding a nucleophilic domain appears to be required for efficient toxin binding.


Space-time clustering of Hodgkin's Disease in parts of the UK, 1984-1993.
The object of this study was to examine cases of Hodgkin's Disease (HD) for evidence of space-time clustering of onsets by age group, sex and disease subtype. Data comprised 2024 cases of HD aged 0-79 years arising throughout the period 1984 to 1993 in the areas covered by a specialist population based register of leukaemias and lymphomas. Knox space-time analysis was used separately for 3 different age groups: childhood (0-14 years), young adult (15-34 years) and older adults (35-79 years); for adult cases separate analysis was carried out by sex and for the nodular and non-nodular sclerosing subtypes. Results showed that space-time clustering of onsets was limited to the nodular sclerosing cases. It was more prominent in young adult nodular sclerosing cases aged 15-34 years (particularly females) diagnosed in the period 1984-88, than in those diagnosed in 1989-93. We conclude that clustering may provide further evidence that an infectious process is involved in the aetiology of young adult nodular sclerosing cases of HD.


Influence of Macromolecular Architecture on the Optical and Humidity-Sensing Properties of Poly(N,N-Dimethylacrylamide)-Based Block Copolymers.
The influence of the macromolecular architecture of block copolymers containing poly(N,N-dimethyl acrylamide) (PDMA) on the optical characteristics and sensing properties of corresponding thin films is discussed. Series of hydrophilic PDMA-based copolymers of different chemical composition and chain architecture such as triblock, star-shaped, and branched were synthesized. The copolymers were characterized using conventional spectroscopic techniques as well as methods for characterization of copolymer macromolecular characteristics in solution, namely size-exclusion chromatography and static light scattering. Thin films of the copolymers of nanometer scale thickness were deposited on silicon substrates by the spin-coating method. The refractive index and extinction coefficient of the copolymer films were calculated from the reflectance spectra by using non-linear curve fitting methods and the composition-structure-optical properties relationships were evaluated. Humidity-sensing properties of the films were studied by measuring reflectance spectra of the films at a relative humidity range from 5 to 95%RH. The implementation of the copolymer films as optical sensors of humidity is justified and discussed.


The dark lymph node sign on magnetic resonance imaging: a novel finding in patients with sarcoidosis.
The purpose of this study was to describe a characteristic magnetic resonance imaging (MRI) appearance of lymphadenopathy in sarcoidosis--the dark lymph node sign (DLNS)--and to determine its prevalence in a retrospective review of cardiopulmonary MRI examinations obtained in patients with sarcoidosis. Fifty-one adult patients with a clinical history of sarcoidosis were evaluated with thoracic MRI during a 15-month span; 29 patients were men, and 22 patients were women. The average age of patients was 53.7±11.2 years. Patients were considered to have the DLNS on MRI if mediastinal or hilar lymph nodes demonstrated internal low intensity with a peripheral circumferential rim of hyperintensity (relative to paraspinal muscle) on post-gadolinium volume-interpolated 3-dimensional gradient echo (3D-GRE/VIBE) and fat-saturated T2-weighted fast spin echo (T2-FSE/BLADE) sequences. Univariate analyses and a logistic regression were used to determine how variables of interest related to the DLNS. Of the 51 patients with sarcoidosis, 49% (25 patients) demonstrated the DLNS. Nodal calcification was present on computed tomography in 45.7% (16/35) of patients with computed tomography scans obtained within 90 days of MRI. The DLNS sign was not more common in those with nodal calcification. When the DLNS occurred in conjunction with calcified nodes, the extent of hypointensity on MRI was not strictly limited to the calcified portions of the lymph node in 71.4% (5/7) of such cases. The DLNS is commonly present on MRI examinations of patients with sarcoidosis, occurring in approximately half of the participants in our study.


Simulation study of surface-plasmon-resonance electro-optic light modulator based on a polymer grating coupler.
An electro-optic light modulator design based on a grating-coupled surface-plasmon-resonance structure is numerically investigated using finite-difference time-domain simulation. The thickness effect of a dielectric layer acting as a waveguide on metal structures is investigated. The results show that the new structure has much higher modulation index than the conventional one and can implement an electro-optic modulator with low operating voltage.


Electrochemical CO2 Reduction Catalyzed by Binuclear LRe2(CO)6Cl2 and LMn2(CO)6Br2 Complexes with an Internal Proton Source.
The synthesis of certain commodity chemicals, e.g., methanol and acetic acid, relies on CO, which is currently mainly produced by the combustion of carbon or natural gas. Photo- or electrochemical conversion of atmospheric CO2 to CO represents an attractive alternative strategy as this approach is carbon-neutral. Such photo- or electrochemically formed CO can also be used in the Fischer-Tropsch process forming liquid hydrocarbons for energy storage applications. The multiple electroreduction of CO2 is preferably coupled with proton transfer steps as this requires less energy than the single outer-sphere 1e- reduction of CO2.In 1984 and 2011, it was shown that [(Lbpy)Re(CO)3Cl] (1) and [(Lbpy)Mn(CO)3Br] (2), respectively, mediate the electrochemical 2e-/2H+ reduction of CO2 forming CO and water (Lbpy = 2,2'-bipyridine). Since proton management is crucial for catalysis, recently the impact of internal proton sources close to the axial position in such complexes has been investigated. However, binuclear complexes have been used rarely as mediators although it has been shown very early for 1 that electron management is also important: the 2e-/2H+ reduction pathway with 1 exhibits a higher reaction rate than going via the singly reduced species, though the pathway requires a higher overpotential. In this Account, we focus on recent developments of binuclear LMn2(CO)6 and LRe2(CO)6 mediators with an internal phenol group in the electroreduction of CO2. In contrast to mononuclear derivatives, for which the impact of the internal proton source on catalysis is very diverse, we always observed a higher reaction rate and for the Mn complexes also a lower overpotential with the binuclear complexes compared to the mononuclear variants. Spectroscopic, electrochemical, and computational studies on the mono- and binuclear complexes shed light on their reactivity under reductive conditions, elucidated the structure of reduced species, unraveled the kinetics for catalytically productive and unproductive (side) reactions, and allowed us to derive some hypothesis on the CO2 reduction mechanism. Finally, I emphasize that the electrohydrogenation of the polar double bonds by the binuclear complex LMn2(CO)6 with a central phenol unit is not restricted to CO2 but is also applicable to organic compounds with C═O bonds.


Mutant huntingtin alters MAPK signaling pathways in PC12 and striatal cells: ERK1/2 protects against mutant huntingtin-associated toxicity.
Huntington's disease (HD) is a devastating neurodegenerative disorder caused by an expanded polyglutamine (polyQ) tract within the huntingtin protein (Htt). Identifying the pathways that are altered in response to the mutant protein is crucial for understanding the cellular processes impacted by the disease as well as for the rational development of effective pharmacological interventions. Here, expression profiling of a cellular HD model identifies genes that implicate altered mitogen-activated protein kinase (MAPK) signaling. Targeted biochemical studies and pharmacological modulation of these MAPK pathways suggest that mutant Htt affects signaling at upstream points such that both ERK and JNK are activated. Modulation of the ERK pathway suggests that this pathway is associated with cell survival, whereas inhibition of JNK was found to effectively suppress pathogenesis. These studies suggest that pharmacological intervention in MAPK pathways, particularly at the level of ERK activation, may be an appropriate approach to HD therapy.


Bifunctional liposomes reduce the chemotherapy resistance of doxorubicin induced by reactive oxygen species.
Doxorubicin (DOX) liposome is a widely used nano-medicine for colorectal cancer treatment. However, doxorubicin therapy increases the level of reactive oxygen species (ROS) in tumor cells, such as hydrogen peroxide (H2O2), which can stabilize hypoxia-inducible-factor-1α (HIF-1α). In a tumor hypoxic microenvironment, HIF-1 can up-regulate tumor-resistance related proteins, including P-glycoprotein (P-gp), glucose transporter 1 (GLUT-1), and matrix metalloproteinase 9 (MMP-9), leading to tumor tolerance to chemotherapy. The functional inhibition of HIF-1 can overcome this resistance and enhance the efficacy of tumor therapy. Here, we encapsulated one of the most effective HIF-1 inhibitors, acriflavine (ACF), and DOX in liposomes (DOX-ACF@Lipo) to construct bifunctional liposomes. ACF and DOX, released from DOX-ACF@Lipo, could effectively suppress the function of HIF-1 and the process of DNA replication, respectively. Consequently, the bifunctional liposome has great potential to be applied in clinics to overcome chemotherapy resistance induced by hypoxia.


Inflammatory mediators involved in antigen-induced airway microvascular leakage in guinea pigs.
Antigen challenge of ovalbumin (OA)-sensitized guinea pigs results in significant (p less than 0.05) increases in vascular permeability to Evans blue (EB) dye in the airways, esophagus, and bladder. Mean values +/- SEM in ng EB/mg wet weight tissue for unsensitized versus sensitized animals were: trachea, 23.6 +/- 6.6 versus 92.5 +/- 11.1; main bronchi, 31.1 +/- 12.2 versus 153.1 +/- 14.9; "central" intrapulmonary airways (ipa), 34.6 +/- 11.2 versus 101.3 +/- 6.2; and "peripheral" ipa, 26.2 +/- 6.8 versus 93.5 +/- 13.6. We investigated the involvement of several mediators of inflammation in this process. FPL 55712, a sulfidopeptide leukotriene receptor antagonist, caused significant inhibition of leakage in trachea (to 55.1 +/- 9.8) and main bronchi (91.7 +/- 15.8). Blockade of the cyclooxygenase and lipoxygenase pathways with BW 755C, but not of the cyclooxygenase pathway alone with indomethacin, also significantly reduced EB dye extravasation in trachea (55.1 +/- 18.0), main bronchi (71.7 +/- 23.0), and "central" ipa (62.7 +/- 16.4). The histamine antagonists, chlorpheniramine and cimetidine, only inhibited microvascular leakage in main bronchi (94.4 +/- 20.0). PAF-receptor blockade with the ginkgolide mixture BN 52063 had no effect. Nedocromil sodium, a mast cell stabilizer and an inhibitor of inflammatory cell activation, caused significant inhibition throughout the airways: trachea, 50.4 +/- 10.6; main bronchi, 72.0 +/- 15.3; "central" ipa 61.0 +/- 8.6; "peripheral" ipa 41.9 +/- 12.2. Thus, histamine and lipoxygenase products (in particular, leukotrienes), but not PAF, may mediate the antigen-induced increase in vascular permeability to different degrees in differing regions of the respiratory tract in guinea pigs.


Apigenin attenuates 2-deoxy-D-ribose-induced oxidative cell damage in HIT-T15 pancreatic β-cells.
Glucose toxicity contributes to progressive β-cell failure and the development of overt diabetes. Oxidative stress is an important aspect of glucose toxicity in pancreatic β-cells. We investigated whether the flavonoid apigenin protects pancreatic β-cells from 2-deoxy-D-ribose (dRib)-induced oxidative cell damage. HIT-T15 pancreatic β-cells were cultured with or without apigenin in the presence of dRib. Time- and dose-dependent cell viability was monitored using a cell counting kit (CCK-8), while the induction of apoptosis was analyzed using a cell death enzyme-linked immunosorbent assay (ELISA) kit. Mitochondrial membrane potential (ΔΨ(m)) was determined using the JC-1 kit. Intracellular oxidative stress was measured by fluorometric analysis of DCFH oxidation using 2',7'-dichlorofluorescin diacetate (DCFH-DA) as the probe. In addition, the DNA binding activity of the oxidative stress-related transcriptional factors nuclear factor-κB (NF-κB) and activator protein 1 (AP-1) were analyzed. dRib reduced cell survival and ΔΨ(m), while it markedly increased intracellular levels of reactive oxygen species (ROS), apoptosis, and the activity of the oxidative stress-related transcription factors NF-κB and AP-1. However, pretreatment of cells with apigenin attenuated all the dRib-induced effects. The anti-oxidants, N-acetyl-L-cysteine (NAC) and alpha lipoic acid (ALA), also prevented both dRib-induced oxidative damage and activation of NF-κB and AP-1. Taken together, these results suggest that apigenin attenuates dRib-induced cell damage in pancreatic β-cells via oxidative stress-related signaling.


Eye mucosa: an efficient vaccine delivery route for inducing protective immunity.
The external part of the eye shares mucosa-associated common characteristics and is an obvious entry site for foreign Ags. We assessed the potential of eyedrop vaccination for effective delivery of vaccines against viral or bacterial infection in mice. Both OVA-specific IgG Ab in serum and IgA Ab in mucosal compartments were induced by eyedrops of OVA with cholera toxin (CT). Eyedrop vaccination of influenza A/PR/8 virus (H1N1) induced both influenza virus-specific systemic and mucosal Ab responses and protected mice completely against respiratory infection with influenza A/PR/8 virus. In addition, eyedrop vaccination of attenuated Salmonella vaccine strains induced LPS-specific Ab and complete protection against oral challenge of virulent Salmonella. Unlike with the intranasal route, eyedrop vaccinations did not redirect administered Ag into the CNS in the presence of CT. When mice were vaccinated by eyedrop, even after the occlusion of tear drainage from eye to nose, Ag-specific systemic IgG and mucosal IgA Abs could be induced effectively. Of note, eyedrops with OVA plus CT induced organogenesis of conjunctiva-associated lymphoid tissue and increased microfold cell-like cells on the conjunctiva-associated lymphoid tissue in the nictitating membrane on conjunctiva, the mucosal side of the external eye. On the basis of these findings, we propose that the eyedrop route is an alternative to mucosal routes for administering vaccines.


Pain in children with juvenile rheumatoid arthritis: a descriptive study.
The purpose of this study is to describe the pain experienced by children with juvenile rheumatoid arthritis. A patient sample composed of 33 children, 7-16 y of age, with juvenile rheumatoid arthritis was included in the study. The children, their parents, and the attending rheumatologist assessed the child's pain using the Abu-Saad pediatric pain assessment tool. In addition, a disease activity index was used by the physician. All children reported pain. This was most often described as hurting, stinging, warm, and uncomfortable. Significant correlations were found for present pain between the child, parent, and physician. The worst pain of the child for the previous week, and not the present pain, correlated with the disease activity as rated by the physician. These findings indicate a need for pain assessment in patients with juvenile rheumatoid arthritis and emphasize the importance of longitudinal studies in this area.


Pharmacokinetics of total and unbound darunavir in HIV-1-infected pregnant women.
To describe the pharmacokinetics of darunavir in pregnant HIV-infected women in the third trimester and post-partum. This was a non-randomized, open-label, multicentre, Phase IV study in HIV-infected pregnant women recruited from HIV treatment centres in Europe. HIV-infected pregnant women treated with darunavir (800/100 mg once daily or 600/100 mg twice daily) as part of their combination ART were included. Pharmacokinetic curves were recorded in the third trimester and post-partum. A cord blood sample and maternal sample were collected. The study is registered at ClinicalTrials.gov under number NCT00825929. Twenty-four women were included in the analysis [darunavir/ritonavir: 600/100 mg twice daily (n=6); 800/100 mg once daily (n=17); and 600/100 mg once daily (n=1)]. Geometric mean ratios of third trimester versus post-partum (90% CI) were 0.78 (0.60-1.00) for total darunavir AUC0-tau after 600/100 mg twice-daily dosing and 0.67 (0.56-0.82) for total darunavir AUC0-tau after 800/100 mg once-daily dosing. The unbound fraction of darunavir was not different during pregnancy (12%) compared with post-partum (10%). The median (range) ratio of darunavir cord blood/maternal blood was 0.13 (0.08-0.35). Viral load close to delivery was <300 copies/mL in all but two patients. All children were tested HIV-negative and no congenital abnormalities were reported. Darunavir AUC and Cmax were substantially decreased in pregnancy for both darunavir/ritonavir regimens. This decrease in exposure did not result in mother-to-child transmission. For antiretroviral-naive patients, who are adherent, take darunavir with food and are not using concomitant medication reducing darunavir concentrations, 800/100 mg of darunavir/ritonavir once daily is adequate in pregnancy. For all other patients 600/100 mg of darunavir/ritonavir twice daily is recommended during pregnancy.


Morphology of Marburg virus NP-RNA.
When Marburg virus (MBGV) nucleoprotein (NP) is expressed in insect cells, it binds to cellular RNA and forms NP-RNA complexes such as insect cell-expressed nucleoproteins from other nonsegmented negative-strand RNA viruses. Recombinant MBGV NP-RNA forms loose coils that resemble rabies virus N-RNA. MBGV NP monomers are rods that are spaced along the coil similar to the nucleoprotein monomers of the rabies virus N-RNA. High salt treatment induces tight coiling of the MBGV NP-RNA, again a characteristic observed for other nonsegmented negative-strand virus N-RNAs. Electron microscopy of fixed Marburg virus particles shows that the viral nucleocapsid has a smaller diameter than the free, recombinant NP-RNA. This difference in helical parameters could be caused by the interaction of other viral proteins with the NP-RNA. A similar but opposite phenomenon is observed for rhabdovirus nucleocapsids that are condensed by the viral matrix protein upon which they acquire a larger diameter. Finally, there appears to be an extensive and regular protein scaffold between the viral nucleocapsid and the membrane that seems not to exist in the other negative-strand RNA viruses.


Approaching pulmonary fat embolism on postmortem computed tomography.
Pulmonary fat embolism (PFE) is a relevant diagnosis playing a role as a sign of vitality or a cause of death. Its severity is assessed according to histological grading systems like that of Falzi. The aim of this study was to determine the utility of unenhanced postmortem computed tomography (PMCT) for PFE diagnosis based on the detection of fat layers. Consecutive cases with PMCT and autopsy were studied retrospectively. The case group consisted of cases with positive PFE, and the control group included cases with negative PFE. Three observers independently assessed PMCT data for fat layers in the pulmonary trunk and the right and left pulmonary artery. For cases with fat layers, autopsy protocols were assessed for the cause of death, relation to trauma, and undertaken resuscitation measures. Eight hundred thirty cases were included: 366 PFE positive cases (144 of Falzi grade 1, 63 of 1.5, 99 of 2, 28 of 2.5, and 32 of 3) and 464 PFE negative cases. Interrater reliabilities varied between substantial and almost perfect, and discrepancies were solved according to majority. Eighteen cases showed fat layers on PMCT (2 controls-traumatic instantaneous deaths-, 16 PFE positive cases). PMCT showed low sensitivity but high specificity for PFE diagnosis. The layers were located at the same position in the pulmonary trunk directly adjacent to the pulmonary valve distal to the right ventricle. Fat layer on PMCT is a rare finding but relates to PFE diagnosis, especially of severe histological grade. It is to be expected in a typical position within the pulmonary trunk.


Use of mean platelet volume in the assessment of intrauterine infection in newborns with combined thrombocytopenia and leukopenia at birth.
Objective: Intrauterine stress can be reflected on hematological disturbance at birth. Thrombocytopenia and leukopenia may be a result of exposure to maternal hypertensive disorders but may also indicate fetal inflammatory response to intrauterine infection, prompting empiric antibiotics use during the initial assessment after birth. Emerging data suggest long-term adverse health outcomes associated with antibiotics exposure early in life. In this report, we sought to assess the use of mean platelet volume (MPV) at birth in predicting fetal inflammatory response in newborns with combined thrombocytopenia and leukopenia.Materials and methods: This is a retrospective study from a single academic medical center. Data were collected prospectively on all newborns with thrombocytopenia and leukopenia within the first 24 h of life. The primary outcome was a composite of fetal tachycardia, premature preterm rupture of membrane with preterm labor, and histological evidence of chorioamnionitis/funisitis/villitis on placental pathology reports evaluated using a multiple logistical regression analysis.Results: The prevalence of combined thrombocytopenia and leukopenia was 5.8% (99 out of 1693 newborns) during the study period. The prevalence was highly associated with gestational age (R2 = 0.873). Twenty-four (32.4%) had abnormal MPV values at birth, defined as MPV > 9 or < 7 fL. Newborns with abnormal MPV had lower platelet counts and were more likely to have I:T ratio ≥0.2. In a univariate analysis, abnormal MPV (OR: 6.205, 95% CI: 1.923-20.022, p = .002), I:T ratio ≥0.2 (OR: 8.462, 95% CI: 1.396-51.281, p = .02), and platelet counts (OR: 98.4, 95% CI: 96.9%-99.9%, p = .035) were each significantly associated with a positive composite outcome. In a multivariate analysis, only abnormal MPV remained significantly associated with an increased likelihood of having a positive composite outcome, with an OR of 3.922 (95% CI: 1.094-14.06, p = .036).Conclusions: MPV may be a more reliable marker than I:T ratio ≥0.2 for fetal inflammatory response in newborns with combined thrombocytopenia and leukopenia during the initial assessment of intrauterine infection. Future prospective studies are required to confirm findings from this report.


Selection for high gamete encounter rates explains the success of male and female mating types.
Sexual reproduction occurs in many small eukaryotes by fusion of similar gametes (isogamy). In the absence of distinguishable sperm and eggs, male and female mating types are missing. However, species with distinct males and females have so prospered that almost all familiar plants and animals have these mating types. Why has sexual reproduction involving sperm and eggs been so successful? An answer is obtained by considering physical limitations on encounter rates between gametes. A biophysical model based on well-established relationships produces fitness landscapes for the evolution of gamete size and energy allocation between motility and pheromone production. These landscapes demonstrate that selection for high gamete encounter rates favors large, pheromone-producing eggs and small, motile sperm. Thus, broadcast-spawning populations with males and females can reproduce at lower population densities and survive under conditions where populations lacking males and females go extinct. It appears that physical constraints on gamete encounter rates are sufficient to explain the first two steps in the isogamy-->anisogamy-->oogamy-->internal fertilization evolutionary sequence observed in several lineages of the eukaryotes. Unlike previous models, assumptions concerning zygote fitness or decreasing speed of swimming with increasing gamete size are not required.


Enhancing Fluoride: Clinical Human Studies of Alternatives or Boosters for Caries Management.
Dental caries remains a major public health problem, especially for certain high-risk population groups. The goal of this study was to assess the evidence regarding strategies meant to be used as alternatives or booster/supplements to fluoride for caries prevention and management. Articles were selected for inclusion if they had a prospective longitudinal design, with a fluoride control arm, and were conducted in human subjects. Of the included studies, 7/18 studies on calcium-based strategies favored the test product (the majority of studies included exposure of fluoride in all groups). All the arginine studies (8/8) included a combination of arginine and a calcium base, and concluded that this has the potential to significantly boost the performance of fluoride. The remaining included studies focused on the addition of microbial-related strategies to a fluoride-containing vehicle (2 xylitol studies and 1 study using a probiotic milk), and all favored the combination as a booster to fluoride. Thus, the current study did not identify evidence for any strategy to effectively be used as a substitute or alternative to fluoride, but identified some consistent evidence derived from the use of prebiotic strategies (primarily from use of arginine combined with calcium) to support their potential use to boost the mechanism of action of fluoride. Thus, fluoride-based strategies remain the standard for caries prevention and management, with some evidence that boosting the effects of fluoride by the use of prebiotic strategies is a promising possibility.


miR-125a-5p regulates megakaryocyte proplatelet formation via the actin-bundling protein L-plastin.
There is heritability to interindividual variation in platelet count, and better understanding of the regulating genetic factors may provide insights for thrombopoiesis. MicroRNAs (miRs) regulate gene expression in health and disease, and megakaryocytes (MKs) deficient in miRs have lower platelet counts, but information about the role of miRs in normal human MK and platelet production is limited. Using genome-wide miR profiling, we observed strong correlations among human bone marrow MKs, platelets, and differentiating cord blood-derived MK cultures, and identified MK miR-125a-5p as associated with human platelet number but not leukocyte or hemoglobin levels. Overexpression and knockdown studies showed that miR-125a-5p positively regulated human MK proplatelet (PP) formation in vitro. Inhibition of miR-125a-5p in vivo lowered murine platelet counts. Analyses of MK and platelet transcriptomes identified LCP1 as a miR-125a-5p target. LCP1 encodes the actin-bundling protein, L-plastin, not previously studied in MKs. We show that miR-125a-5p directly targets and reduces expression of MK L-plastin. Overexpression and knockdown studies show that L-plastin promotes MK progenitor migration, but negatively correlates with human platelet count and inhibits MK PP formation (PPF). This work provides the first evidence for the actin-bundling protein, L-plastin, as a regulator of human MK PPF via inhibition of the late-stage MK invagination system, podosome and PPF, and PP branching. We also provide resources of primary and differentiating MK transcriptomes and miRs associated with platelet counts. miR-125a-5p and L-plastin may be relevant targets for increasing in vitro platelet manufacturing and for managing quantitative platelet disorders.


Response of triiodothyronine-dependent enzyme activities to insulin-like growth factor I and growth hormone in cultured rat hepatocytes.
Triiodothyronine (T3) is involved in the regulation of the growth hormone-insulin-like growth factor I (GH-IGF-I) axis. In this study we investigated the effect of GH and IGF-I on the metabolic response of T3 in target tissues by evaluating the activity of two T3-dependent liver enzymes: mitochondrial alpha-glycerophosphate dehydrogenase (alpha-GPD) and cytosolic malic enzyme (ME) in rat hepatocytes in primary culture. Growth hormone (35 nmol/l) as well as IGF-I (0.5 mumol/l) reduced alpha-GPD and ME activities (p < 0.01) compared to the control group. Timecourse studies indicated that IGF-I (1.5 mumol/l) significantly decreased alpha-GPD and ME activities (P < 0.01) after 24 h, whereas the effect of GH (35 nmol/l) was recorded only after 36 h (p < 0.01). This delayed effect of GH compared to IGF-I suggested the possibility that the effect of GH could be mediated by IGF-I synthesis. To test this hypothesis, the effect of GH on the two enzyme activities was studied in the presence of anti-IGF-I antibodies. A gradual recovery of alpha-GPD and ME activities (p < 0.01) was observed in the presence of GH (35 nmol/l) plus increasing concentrations of anti-IGF-I antiserum. The maximal alpha-GPD and ME activities attained after the incubation of the liver cells with 1 mumol/l T3, a concentration high enough to fully saturate the nuclear T3 receptors for 24 h, were lowered significantly by 1.0 mumol/l IGF-I (p < 0.01). This finding suggests that the IGF-I effect might be independent of the saturation of the nuclear T3 receptors. In conclusion, in cultured rat hepatocytes, GH and IGF-I reduced the metabolic response of T3 evaluated by two liver T3-dependent enzyme activities. The effect of GH was mediated at least in part by IGF-I.


Third-Generation Minimally Invasive Chevron Akin Osteotomy for Hallux Valgus.
Multiple operative techniques have been developed for hallux valgus with varying success. The most recent developments in minimally invasive surgery have evolved into the third-generation minimally invasive chevron Akin (MICA) osteotomy. Good results have been shown from originator centers, but this is one of the first series from a nonoriginator center, and the first to use a validated patient-reported outcome measure. Forty consecutive patients undergoing third-generation MICA for hallux valgus were included. Primary outcome measures included Manchester-Oxford Foot Questionnaire (MOXFQ) and American Orthopaedic Foot & Ankle Society (AOFAS) scores and Coughlin satisfaction rates at 12 months. Secondary outcome measures included radiographic parameters, complications, and recurrence rates. At 12 months, the MOXFQ score improved from 58 to 10 and the AOFAS score improved from 48 to 93, with 70% of patients reporting excellent outcomes and 30% good ones. Two cases started as mild, 29 cases as moderate, and 9 cases as severe as defined by radiographic criteria. Hallux valgus angles improved from 32 degrees to 12 degrees, and intermetatarsal angles improved from 13 degrees to 7 degrees. There were 4 cases of Akin screw removal for soft tissue irritation. There were no other complications, including recurrence. The third-generation MICA technique was a safe and effective approach to treating hallux valgus. Further research should focus on long-term outcomes and comparative data with other commonly performed operative techniques. Level IV, case series.


Development and Performance Evaluation of a Novel Ancestry Informative DIP Panel for Continental Origin Inference.
Ancestry informative markers (AIMs) are useful to infer individual biogeographical ancestry and to estimate admixture proportions of admixed populations or individuals. Although a growing number of AIM panels for forensic ancestry origin analyses were developed, they may not efficiently infer the ancestry origins of most populations in China. In this study, a set of 52 ancestry informative deletion/insertion polymorphisms (AIDIPs) were selected with the aim of effectively differentiate continental and partial Chinese populations. All of the selected markers were successfully incorporated into a single multiplex PCR panel, which could be conveniently and efficiently detected on capillary electrophoresis platforms. Genetic distributions of the same 50 AIDIPs in different continental populations revealed that most loci showed high genetic differentiations between East Asian populations and other continental populations. Population genetic analyses of different continental populations indicated that these 50 AIDIPs could clearly discriminate East Asian, European, and African populations. In addition, the 52 AIDIPs also exhibited relatively high cumulative discrimination power in the Eastern Han population, which could be used as a supplementary tool for forensic investigation. Furthermore, the Eastern Han population showed close genetic relationships with East Asian populations and high ancestral components from East Asian populations. In the future, we need to investigate genetic distributions of these 52 AIDIPs in Chinese Han populations in different regions and other ethnic groups, and further evaluate the power of these loci to differentiate different Chinese populations.


Preparedness components of health systems in the Eastern Mediterranean Region for effective responses to dust and sand storms: a systematic review.
Background: Dust and Sand Storm (DSS), according to estimates by global reports, will increase dramatically in the Eastern Mediterranean Region (EMR). Numerous health problems caused by DSS will be severely affected regions and vulnerable groups. This study aimed to identify the components of the preparedness of health systems for the DSS phenomenon in EMR. Methods: In this systematic review, the peer-reviewed papers in four electronic databases, including Medline through PubMed, Scopus, ISI Web of Science and the Cochrane library, as well as available grey literature, were searched and selected. The research process was carried out by including papers whose results were related to the potential health effects caused by desert dusts in EMR. Was used the combination of three groups of keywords: the exposure factor, health effects as outcomes, and the countries located in EMR. The focus was on the PRISMA checklist, with no time limitations until December 2017. Finally, through 520 related citations, 30 articles were included. Descriptive and thematic content analyses were evaluated. Results: The preparedness components were divided into three and ten main categories and subcategories, respectively. The three categories covered the areas of DSS hazard identification, planning and policy-making, and risk assessment. Conclusions: Recognition of the health system preparedness factors for DSS in EMR will help policy-makers and managers perform appropriate measures when dealing with this hazard. More studies should be conducted to understand these factors in other parts of the world. Registration: PROSPERO registration number CRD42018093325.


The incidence and risk factors of hypofibrinogenemia in cardiovascular surgery.
Cardiovascular surgery often causes massive bleeding due to coagulopathy, with hypofibrinogenemia being a major causative factor. We assessed the intraoperative incidence of hypofibrinogenemia and explored predictors of hypofibrinogenemia. The intraoperative serum fibrinogen level (SFL) was routinely measured in 872 consecutive patients [mean age: 66.9 ± 13.3 years; 598 men (68.6%)] undergoing cardiovascular surgery from July 2013 to November 2016 at Nagoya University Hospital. There were 275 aortic surgeries, 200 cases of coronary artery bypass grafting (CABG), 334 valvular surgeries and 63 other surgeries. We estimated hypofibrinogenemia incidence (intraoperative lowest SFL ≤ 150 mg/dL) and identified its predictors by a logistic regression analysis. The average intraoperative lowest SFL of all cases, aortic surgery, CABG and valvular surgery was 185 ± 71, 156 ± 65, 198 ± 69 and 198 ± 68 mg/dL, respectively. Aortic surgery had a significantly lower intraoperative lowest SFL than CABG (p < 0.001) and valvular surgery (p < 0.001). The incidence of hypofibrinogenemia was 32.8%, 50.2%, 26.5% and 22.8% in all cases, aortic surgery, CABG and valvular surgery, respectively. The predictors of hypofibrinogenemia were the preoperative SFL, re-do surgery and perfusion time. A receiver operating characteristics curve analysis showed that the best preoperative SFL cutoff value for predicting hypofibrinogenemia was 308.5 mg/dL. Assuming preoperative SFL 300 mg/dL as the cutoff, the odds ratio for hypofibrinogenemia was 7.22 (95% confidence interval 5.26-9.92, p < 0.001). The incidence of hypofibrinogenemia in aortic surgery was high. The preoperative SFL, re-do surgery and perfusion time were identified as predictors for hypofibrinogenemia. Intraoperative measurement of SFL is important for detecting hypofibrinogenemia and applying appropriate and prompt transfusion treatment.


A case of long-term survival after multimodal local treatments of intramedullary spinal cord metastasis of squamous cell lung cancer.
Intramedullary spinal cord metastasis of non-small cell lung cancer is rare, and it has a short prognosis. We report a 53-year-old man diagnosed with cT4N0M0, stage IIIA squamous cell lung cancer. Ten months after left pneumonectomy (pT4N0M0), an intramedullary spinal cord tumor developed at the axis level. The intramedullary spinal cord tumor was resected, and he was diagnosed with metastatic squamous cell lung cancer. Radiotherapies and another tumor resection were conducted, as he had a good performance status and the discrete lesion was associated with the risk of brain stem compression. Multimodal local treatments for intramedullary spinal cord metastasis caused the tumor to shrink, and he lived for 25 months after the spinal metastasis occurred.


Clinical presentation of exclusive cystic fibrosis lung disease.
The diagnosis of cystic fibrosis (CF) is based on the occurrence of two mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene and on assays that measure the basic defect of abnormal chloride transport in the affected organs. However, in cases of atypical CF not all diagnostic tests may be positive. We present a patient with an atypical CF phenotype in whom the only presenting symptom was severe CF-like lung disease substantiated by an abnormal nasal potential difference. Genetic analysis showed that the patient was a symptomatic heterozygote, which suggests that one lesion in the CFTR gene may be sufficient to cause CF-like lung disease.


A new statistical detector for DWT-based additive image watermarking using the Gauss-hermite expansion.
Traditional statistical detectors of the discrete wavelet transform (DWT)-based image watermarking use probability density functions (PDFs) that show inadequate matching with the empirical PDF of image coefficients in view of the fact that they use a fixed number of parameters. Hence, the decision values obtained from the estimated thresholds of these detectors provide substandard detection performance. In this paper, a new detector is proposed for the DWT-based additive image watermarking, wherein a PDF based on the Gauss-Hermite expansion is used, in view of the fact that this PDF provides a better statistical match to the empirical PDF by utilizing an appropriate number of parameters estimated from higher-order moments of the image coefficients. The decision threshold and the receiver operating characteristics are derived for the proposed detector. Experimental results on test images demonstrate that the proposed watermark detector performs better than other standard detectors such as the Gaussian and generalized Gaussian (GG), in terms of the probabilities of detection and false alarm as well as the efficacy. It is also shown that detection performance of the proposed detector is more robust than the competitive GG detector in the case of compression, additive white Gaussian noise, filtering, or geometric attack.


SPRC Suppresses Experimental Periodontitis by Modulating Th17/Treg Imbalance.
Object: The aims of the study were to explore the protective effects of S-propargyl-cysteine (SPRC) on periodontitis and to determine the underlying mechanisms. Methods: A rat periodontitis model was constructed by injecting LPS and SPRC (0, 25, and 50 mg/kg/d) was administered intraperitoneally. H2S and CSE level were detected. The alveolar bone level was evaluated by micro-CT, HE staining and methylene blue staining analysis. Inflammation-related factors, Treg and Th17 cells were detected by immunohistochemistry, RT-PCR, immunofluorescence, Western blot and flow cytometry. Phosphorylation levels of ERK1/2 and CREB were analysed. Results: The administration of SPRC significantly increased the expression of CSE in the gingival tissue and the concentration of endogenous H2S in the peripheral blood. Simultaneously, SPRC significantly inhibited the resorption of alveolar bone based on the H&E staining, micro-CT and methylene blue staining analysis. Compared with the periodontitis group, the levels of IL-17A, IL-10 were downregulated and IL-6,TGF-β1 were upregulated in the SPRC groups. In the SPRC group, the percentage of TH17 cells and the expression of ROR-γt were downregulated, while the percentage of Tregs and the expression of Foxp3 were upregulated accompanied with inhibition of phosphorylation ERK1/2 and CREB. Conclusion: SPRC can prevent the progression of periodontitis by regulating the Th17/Treg balance by inhibition of the ERK/CREB signalling pathway.


The influence of Achilles tendon mechanical behaviour on "apparent" efficiency during running at different speeds.
We investigated the role of elastic strain energy on the "apparent" efficiency of locomotion (AE), a parameter that is known to increase as a function of running speed (up to 0.5-0.7) well above the values of "pure" muscle efficiency (about 0.25-0.30). In vivo ultrasound measurements of the gastrocnemius medialis (GM) muscle-tendon unit (MTU) were combined with kinematic, kinetic and metabolic measurements to investigate the possible influence of the Achilles tendon mechanical behaviour on the mechanics (total mechanical work, WTOT) and energetics (net energy cost, Cnet) of running at different speeds (10, 13 and 16 km h-1); AE was calculated as WTOT/Cnet. GM fascicles shortened during the entire stance phase, the more so the higher the speed, but the majority of the MTU displacement was accommodated by the Achilles tendon. Tendon strain and recoil increased as a function of running speed (P < 0.01 and P < 0.001, respectively). The contribution of elastic energy to the positive work generated by the MTU also increased with speed (from 0.09 to 0.16 J kg-1 m-1). Significant negative correlations (P < 0.01) were observed between tendon work and metabolic energy at each running speed (the higher the tendon work the lower the metabolic demand) and significant positive correlations were observed between tendon work and AE (P < 0.001) at each running speed (the higher the tendon work the higher the efficiency). These results support the notion that the dynamic function of tendons is integral in reducing energy expenditure and increasing the "apparent" efficiency of running.


Mechlorethamine, vincristine, melphalan and prednisolone rescue chemotherapy protocol for resistant feline lymphoma.
Objectives The goals of this retrospective study were to evaluate the use of mechlorethamine, vincristine, melphalan and prednisolone (MOMP) chemotherapy for rescue of feline lymphoma, to describe the protocol's toxicity and to determine prognostic indicators for progression-free survival. Methods The medical records of 12 cats treated with MOMP chemotherapy at the University of Tennessee Veterinary Medical Center between 2007 and 2017 were evaluated. Parameters assessed included lymphoma cell size, anatomical location, number of previous chemotherapy drugs and number of previous rescue protocols received. Chemotherapy-related toxicity was also described. Results Seven of 12 cats responded to this rescue protocol. Three cats experienced complete response and four cats achieved partial response for a median duration of 39 days (range 14-345 days). Cats that achieved complete response had a significantly longer median progression-free survival than cats that did not respond to treatment. Five of 12 cats developed hematologic toxicity (neutropenia) and one cat developed gastrointestinal toxicity. Toxicity was mild in most cases; no cats needed to be hospitalized. Neutropenia was associated with increased progression-free survival. Conclusions and relevance MOMP is a safe and effective rescue chemotherapy protocol for cats with relapsing and refractory lymphoma.


Dopamine and reward: comment on Hernandez et al. (2006).
Many lines of evidence suggest that the dopaminergic projection from the midbrain tegmentum to the forebrain must play a critical role in mediating the behavioral effects of natural and artificial rewards, with brain stimulation reward and addictive drugs included in the latter category. However, a closer look reveals many incongruities. The work of G. Hernandez et al. (2006) resolves several puzzles. It implies that the dopaminergic projection does not carry the signal that encodes the magnitude of a brain stimulation reward. It suggests that the elevation in the tonic levels of dopamine consequent on brain stimulation reward modulates the registration of the magnitude of the reward. This reconciles the psychophysical evidence with the pharmacological, electrophysiological, and anatomical evidence. However, some serious puzzles do remain.


Δ9-Tetrahydrocannabinol-like discriminative stimulus effects of compounds commonly found in K2/Spice.
A number of cannabinoid compounds are being sold in the form of incense as 'legal' alternatives to marijuana. The purpose of these experiments was to determine whether the most common of these compounds have discriminative stimulus effects similar to Δ-tetrahydrocannabinol (Δ-THC), the main active component in marijuana. Locomotor depressant effects of JWH-018, JWH-073, JWH-200, JWH-203, JWH-250, AM-2201, and CP 47,497-C8-homolog were tested in mice. The compounds were then tested for substitution in rats trained to discriminate Δ-THC (3 mg/kg, intraperitoneally). The time course of the peak dose of each compound was also tested. Each of the synthetic cannabinoids dose-dependently decreased locomotor activity for 1-2 h. Each of the compounds fully substituted for the discriminative stimulus effects of Δ-THC, mostly at doses that produced only marginal amounts of rate suppression. JWH-250 and CP 47,497-C8-homolog suppressed response rates at doses that fully substituted for Δ-THC. The time courses varied markedly between compounds. Most of the compounds had a shorter onset than Δ-THC, and the effects of three of the compounds lasted substantially longer (JWH-073, JWH-250, and CP 47,497-C8-homolog). Several of the most commonly used synthetic cannabinoids produce behavioral effects comparable with those of Δ-THC, which suggests that these compounds may share the psychoactive effects of marijuana responsible for abuse liability. The extremely long time course of the discriminative stimulus effects and adverse effects of CP 47,497-C8-homolog suggest that CP 47,497-C8-homolog may be associated with increased hazards among humans.


Antigen-Independent Restriction of Pneumococcal Density by Mucosal Adjuvant Cholera Toxin Subunit B.
For many bacterial respiratory infections, development of (severe) disease is preceded by asymptomatic colonization of the upper airways. For Streptococcus pneumoniae, the transition to severe lower respiratory tract infection is associated with an increase in nasopharyngeal colonization density. Insight into how the mucosal immune system restricts colonization may provide new strategies to prevent clinical symptoms. Several studies have provided indirect evidence that the mucosal adjuvant cholera toxin subunit B (CTB) may confer nonspecific protection against respiratory infections. Here, we show that CTB reduces the pneumococcal load in the nasopharynx, which required activation of the caspase-1/11 inflammasome, mucosal T cells, and macrophages. Our findings suggest that CTB-dependent activation of the local innate response synergizes with noncognate T cells to restrict bacterial load. Our study not only provides insight into the immunological components required for containment and clearance of pneumococcal carriage, but also highlights an important yet often understudied aspect of adjuvants.


Advanced rescue techniques: lessons learned from the collapse of a building in Nairobi, Kenya.
This paper examines the collapse of a five-storey building in Nairobi, Kenya, on 23 January 2006. It draws on reports from local authorities and on debriefings by Israel's Home Front Command (HFC), including information on injury distribution, rescue techniques, and the mode of operation. Most of the 117 people found under the structure were evacuated on the first day to a public hospital, which was overwhelmed by the incident. HFC forces arrived 23 hours after the disaster. At that stage, two people were still buried under the building and special techniques (tunnelling and scalping) were required to secure their evacuation. The two people quickly recovered after a short stay in hospital. Local technology is the preferred option during such events because time is crucial. International cooperation is required when this technology is not available. All of the hospitals in the disaster area, including private facilities, should participate in treating casualties.


A consensus statement for safety monitoring guidelines of treatments for major depressive disorder.
This paper aims to present an overview of screening and safety considerations for the treatment of clinical depressive disorders and make recommendations for safety monitoring. Data were sourced by a literature search using MEDLINE and a manual search of scientific journals to identify relevant articles. Draft guidelines were prepared and serially revised in an iterative manner until all co-authors gave final approval of content. Screening and monitoring can detect medical causes of depression. Specific adverse effects associated with antidepressant treatments may be reduced or identified earlier by baseline screening and agent-specific monitoring after commencing treatment. The adoption of safety monitoring guidelines when treating clinical depression is likely to improve overall physical health status and treatment outcome. It is important to implement these guidelines in the routine management of clinical depression.


Progress in research and application of PLGA embolic microspheres.
Transcatheter arterial chemoembolization was an important method in the treatment of solid cancers and the effectiveness of such treatment depends on the nature of embolic agent. Due to the biocompatibility, controllable degradation rate, and both hydrophobicity-hydrophilicity, researches of application on poly(lactic-co-glycolic acid) (PLGA) in medical practice has been ongoing for more than 40 years. We have seen many benefits for patients in recent years. There were five different methods of preparing micrometer-scale microspheres, and three kinds of PLGA microspheres have been subjected to experimental research or used in clinical applications, namely blank microspheres, drug-loaded microspheres, and radioactive microspheres. Hereby, we reviewed the production and clinical and experimental applications of PLGA microspheres in practice.


Effect of 1.5% sodium hydroxide final concentration on recovery rate of Mycobacterial Species and decontamination of other Bacterial and Fungal contaminants on sputum.
Digestion and decontamination of non-sterile clinical specimens such as sputum are an essential step in the isolation of mycobacteria. Masking of mycobacteria in Mycobacterial growth indicator tube (MGIT) 960 liquid culture system by fungi and bacteria other than mycobacteria is a major problem. To assess the effect of 1.5% sodium hydroxide final concentration on recovery rate of mycobacterial species and decontamination of other bacterial and fungal contaminants from sputum sample. Laboratory based cross sectional study with convenient sampling technique was carried out on subjects referred to the National Tuberculosis Reference Laboratory of Ethiopian Public Health Institute from November 2015 to February 2016. Single morning sputum was collected from each patient and analyzed. A total of 264 subjects were enrolled in the study. The mean age of participant was 31 (SD 20.14 - 41.42) years old. The majority (61%) were male. Increasing the final concentration of NaOH from 1% to 1.5% reduced the contamination rate from 22.4% to 6.8% (P<0.001) without affecting mycobacterial recovery (P=1.00). A total of 26 different species of microbial contaminants were identified as being associated with BACTEC MGIT 960 culture system. Results presented in this study demonstrated that the use of a final concentration of 1.5% NaOH with NALC method aids in reducing culture contamination rate for decontaminating sputum samples referred for tuberculosis culture diagnosis. Among the identified microbial contaminants, the most predominant was coagulase negative Staphylococcus species.


Metal uptake of recombinant cambialistic superoxide dismutase from Propionibacterium shermanii is affected by growth conditions of host Escherichia coli cells.
We constructed the complete nucleotide sequence coding for the cambialistic superoxide dismutase from Propionibacterium shermanii by ligation of a synthetic linker to a polymerase chain reaction amplification product obtained using degenerate primers. We set up an expression system yielding large amounts of recombinant superoxide dismutase in the cytoplasm of Escherichia coli and purified the enzyme from cells grown in a complex medium. The physicochemical properties of the recombinant enzyme were identical to those of the natural protein. Under anaerobic conditions the enzyme produced in an iron-supplemented medium incorporated iron as metal cofactor, while the enzyme purified from cells grown under aerobic conditions contained a variable amount of iron and manganese depending on metal availability. Functional equivalence of the two metals in this superoxide dismutase variant was indicated by independence of enzyme activity from Fe/Mn ratio.


Potent single-domain antibodies that arrest respiratory syncytial virus fusion protein in its prefusion state.
Human respiratory syncytial virus (RSV) is the main cause of lower respiratory tract infections in young children. The RSV fusion protein (F) is highly conserved and is the only viral membrane protein that is essential for infection. The prefusion conformation of RSV F is considered the most relevant target for antiviral strategies because it is the fusion-competent form of the protein and the primary target of neutralizing activity present in human serum. Here, we describe two llama-derived single-domain antibodies (VHHs) that have potent RSV-neutralizing activity and bind selectively to prefusion RSV F with picomolar affinity. Crystal structures of these VHHs in complex with prefusion F show that they recognize a conserved cavity formed by two F protomers. In addition, the VHHs prevent RSV replication and lung infiltration of inflammatory monocytes and T cells in RSV-challenged mice. These prefusion F-specific VHHs represent promising antiviral agents against RSV.


Protein quantification by LC-MS: a decade of progress through the pages of Bioanalysis.
Over the past 10 years, there has been a remarkable increase in the use of LC-MS for the quantitative determination of proteins, and this technique can now be considered an established bioanalytical platform for the quantification of macromolecular drugs and biomarkers, next to the traditional ligand-binding assays. Many researchers have contributed to the field and helped improve both the technical possibilities of LC-MS-based workflows and our understanding of the meaning of the results that are obtained. As a tribute to Bioanalysis, which has published many important contributions, this report gives a high-level overview of the most important trends in the field of protein LC-MS, as published in this journal since its inauguration a decade ago. It describes the major technical developments with regard to sample handling, separation and MS detection of both digested and intact protein analysis. In addition, the relevance of the complex structure and in vivo behavior of proteins is discussed and the effect of protein-protein interactions, biotransformation and the occurrence of isoforms on the analytical result is addressed.


Lyophilized type-I collagen and chronic leg ulcers.
Lyophilized type I collagen (L.C.) can stimulate wound healing by recruiting a number of different cell types (i.e. platelets and macrophages) and proteins (i.e. fibronectin). Platelets and macrophages produce locally-acting growth factors that in turn induce fibroblast and epidermal migration, angiogenesis and increase matrix synthesis. Chronic leg ulcers (C.L.U.) are the end result of microvascular failure owing to ischemia and stasis. When L.C. has been used in the treatment of C.L.U. we have observed that: a) it is significantly more effective in stimulating the healing of chronic venous ulcers when compared to hydrocolloids (p less than .05), the two products being applied upon half of the same ulcer; b) in the treatment of C.L.U. due to arterial obstruction L.C. is more effective than hydrocolloids without achieving statistical significance; c) it is very effective in the treatment of C.L.U. in thalassaemic patients; d) telethermographic studies have demonstrated an increase of blood perfusion and histological studies have shown the stimulation of angiogenesis, fibropoiesis and epidermal growth; e) the application of L.C. determines the maximum obtainable increase also under conditions of proven cicatrization difficulty; and f) enzymatic degradation of L.C. has not promoted any bacterial infection and no local or generalized sensibilization phenomena have been observed. We can conclude that L.C. is a pharmacological approach to wound healing, directly interfering with cellular and non-cellular components, and significantly improves the reparative process when delayed.


Single and double substrate insertion into the Ti=N(alpha) bonds of terminal titanium hydrazides.
Nitriles, CO(2) and isocyanates undergo net single or double insertion reactions into the Ti=N(alpha) multiple bonds of terminal titanium hydrazides. These are the first such examples of this type of reactivity for any transition metal hydrazide complex.


[In vitro and in vivo intestinal absorption of Huanglian decoction in multi-component environment].
The effect of drugs in the body is always inseparable with the dynamic processes of absorption, distribution, metabolism and excretion. For oral drug, absorption, as the first link to enter the body, is particularly valued. Traditional Chinese medicine has multiple components and multiple targets, and the nature of its single composition is different from that in the multicomponent environment. Alkaloids in Huanglian extract was used as the main object in this study to establish an analytical method for determining the content of alkaloids in Huanglian. In addition, the compositions of the Huanglian aqueous extract solution which can be absorbed through intestinal wall into blood, were initially determined by the means of everted gut sac and in intestinal perfusion with venous sampling experiment. This paper can provide data reference and support for the further study on the absorption and metabolism of Huanglian.


Latitudinal patterns of travel among returned travelers with influenza: results from the GeoSentinel Surveillance Network, 1997-2007.
Influenza is a common vaccine-preventable disease among international travelers, but few data exist to guide use of reciprocal hemisphere or out-of-season vaccines. We analyzed records of ill-returned travelers in the GeoSentinel Surveillance Network to determine latitudinal travel patterns in those who acquired influenza abroad. Among 37,542 ill-returned travelers analyzed, 59 were diagnosed with influenza A and 11 with influenza B. Half of travelers from temperate regions to the tropics departed outside influenza season. Twelve travelers crossed hemispheres from one temperate region to another, five during influenza season. Ten of 12 travelers (83%) with influenza who crossed hemispheres were managed as inpatients. Proportionate morbidity estimates for influenza A acquisition were highest for travel to the East-Southeast Asian influenza circulation network with 6.13 (95% CI 4.5-8.2) cases per 1000 ill-returned travelers, a sevenfold increased proportionate morbidity compared to travel outside the network. Alternate hemisphere and out-of-season influenza vaccine availability may benefit a small proportion of travelers. Proportionate morbidity estimates by region of travel can inform pre-travel consultation and emphasize the ease of acquisition of infections such as influenza during travel.


[Cerebral monitoring in anesthesia and intensive care].
To ensure efficient computer-based cerebral monitoring in the ICU and during anaesthesia, sophisticated and sometimes expensive equipment is needed. It is thus all the more important that the clinician should have a good understanding and knowledge of the various methods involved, the origins of the different signals and their relationships, and of the possibilities of artefacts occurring, if the information that this equipment is capable of providing is to be fully and properly utilized. The present paper describes a number of new techniques of cerebral monitoring, and their successful application in the intensive care unit and the operating theatre.


[Computed tomography in the study of gastrointestinal perforation].
Plain radiography is the method of choice in suspected perforating pneumoperitoneum. Nevertheless, especially when air collections are small, the technique must be very accurate, with patient mobilization and long examination times, which may be unfeasible in acute abdomen patients. To overcome these limitations, such cross-sectional imaging methods as US and especially CT are increasingly used. Our series consisted of 38 patients with gastrointestinal tract perforation examined 1990-94; thirty-one of them had surgical confirmation. CT had high sensitivity, demonstrating the presence of free intraperitoneal gas in more patients than conventional radiography (92% vs. 74%). Pneumoperitoneum was depicted between liver surface and anterior abdominal wall in 30 cases, in the subhepatic region in 17, posterior to the abdominal wall at paraumbilical level in 14, between the mesenteric folds in 8, in the pelvis in 7 and in other locations in 11. Extraluminal fluid collections were apparent in 79% of cases and contrast agent collections in 73%. The three most common findings were: intraperitoneal gas, fluid effusion and extraluminal contrast agent leaks (61.5%), gas and effusion (29%) and gas only (16%). The origin of the perforation was demonstrated in 82% of cases and its cause in 37%. CT was particularly useful in the diagnosis of clinically occult, of early and of confined perforations. Small gas bubbles, mild fluid effusion or minimum contrast agent leaks near perforation site are valuable signs. In selected cases CT can play an integrative role, thus improving the diagnostic accuracy of plain films.


Voluntary stopping of eating and drinking: is medical support ethically justified?
Physician-assisted dying has been the subject of extensive discussion and legislative activity both in Europe and North America. In this context, dying by voluntary stopping of eating and drinking (VSED) is often proposed, and practiced, as an alternative method of self-determined dying, with medical support for VSED being regarded as ethically and legally justified. In our opinion, this view is flawed. First, we argue that VSED falls within the concept of suicide, albeit with certain unique features (non-invasiveness, initial reversibility, resemblance to the natural dying process). Second, we demonstrate, on the basis of paradigmatic clinical cases, that medically supported VSED is, at least in some instances, tantamount to assisted suicide. This is especially the case if a patient's choice of VSED depends on the physician's assurance to provide medical support. Thus, for many jurisdictions worldwide, medically supported VSED may fall within the legal prohibitions on suicide assistance. Physicians, lawmakers, and societies should discuss specific ways of regulating medical support for VSED in order to provide clear guidance for both patients and healthcare professionals. Please see related article: http://bmcmedicine.biomedcentral.com/articles/10.1186/s12916-017-0951-0 .


Genetic manipulation of single neurons in vivo reveals specific roles of flamingo in neuronal morphogenesis.
To study the roles of intracellular factors in neuronal morphogenesis, we used the mosaic analysis with a repressible cell marker (MARCM) technique to visualize identifiable single multiple dendritic (MD) neurons in living Drosophila larvae. We found that individual neurons in the peripheral nervous system (PNS) developed clear morphological polarity and diverse dendritic branching patterns in larval stages. Each MD neuron in the same dorsal cluster developed a unique dendritic field, suggesting that they have specific physiological functions. Single-neuron analysis revealed that Flamingo did not affect the general dendritic branching patterns in postmitotic neurons. Instead, Flamingo limited the extension of one or more dorsal dendrites without grossly affecting lateral branches. The dendritic overextension phenotype was partially conferred by the precocious initiation of dorsal dendrites in flamingo mutant embryos. In addition, Flamingo is required cell autonomously to promote axonal growth and to prevent premature axonal branching of PNS neurons. Our molecular analysis also indicated that the amino acid sequence near the first EGF motif is important for the proper localization and function of Flamingo. These results demonstrate that Flamingo plays a role in early neuronal differentiation and exerts specific effects on dendrites and axons.


Molecular diagnosis and biochemical studies of tick-borne diseases (anaplasmosis and babesiosis) in Aberdeen Angus Cattle in New Valley, Egypt.
Anaplasmosis and babesiosis are tick-borne diseases that threaten livestock production with subsequent considerable economic losses. This study was conducted to diagnose Anaplasma and Babesia infection using molecular techniques in imported Aberdeen Angus cattle imported from Uruguay to El-Kharga Oasis in New Valley, Egypt, and to investigate the effects of disease on some serum biochemical and oxidative stress parameters. Blood samples were collected from 31 cattle, 21 diseased and ten apparently normal, of varying ages and sex. The blood was used for the preparation of blood smears, polymerase chain reaction assay, and separation of serum for biochemical investigation. The experimental production farm at the Faculty of Agriculture, New Valley University, was infested with ticks and variable clinical manifestations during the period from December 2017 to March 2018. One calf died of a suspected blood parasite infection. The blood film examination revealed infection by blood parasites in 21 samples. Anaplasma marginale and Babesia bovis were identified in 12 and 14 samples, respectively. A total of 14 samples were examined by polymerase chain reaction (PCR) to make these identifications. Biochemical parameters showed significantly elevated serum alanine aminotransferase, aspartate aminotransferase, total bilirubin (T. Bil), and urea in blood from parasite-infected female cattle and male calves compared with controls. Increased serum total protein, globulin, and creatinine were recorded only in infected female cattle. The blood glucose level was significantly decreased in infected female cattle and male calves compared with controls. Furthermore, albumin and albumin/globulin ratio was significantly reduced in the infected female cattle. Oxidative stress profiles of infected animals showed a significant increase in serum nitric oxide and malondialdehyde, and both total antioxidant capacity and reduced glutathione (GSH) were significantly reduced in comparison with control animals. The incidence of A. marginale and B. bovis infection is high in imported Aberdeen Angus cattle in New Valley Province. PCR methods provide a short-term assessment of disease. An extensive epidemiological survey, employing serology together with molecular genetic methods, monitoring of abundance and distribution of tick vectors, availability of vaccination programs, and tracking of animal transport is also needed for control of blood parasites.


Rheological properties and baking performance of new oat beta-glucan-rich hydrocolloids.
Two new oat beta-glucan hydrocolloids (designated C-trim20 and C-trim30) obtained through a thermal-shearing process were evaluated for their potential use in food products as functional ingredients. Their rheological characteristics were investigated using steady and dynamic shear measurements. Both samples exhibited typical shear-thinning and viscoelastic properties of random coil polysaccharides. The Cross equation was also used to examine the dependence of their apparent viscosity on shear rates. Furthermore, the effects of flour replacement with C-trim20 on the physical, rheological, and sensory properties of cookies were studied. The cookies containing C-trim20 exhibited reduced spreading characteristics compared with the control due to their increased elastic properties. Also, higher water content and water activity were observed in the C-trim20 cookies. However, flour replacement with C-trim20 up to 10% produced cookies with instrumental texture properties similar to those of the control, which was in good agreement with the sensory results.


Expression of the herpes simplex virus type 1 latency-associated transcripts does not influence latency establishment of virus mutants deficient for neuronal replication.
Herpes simplex virus type 1 establishes latency within neurons of the trigeminal ganglion. During latency, viral gene expression is largely restricted to the latency-associated transcripts (LATs), which, whilst not essential for any aspect of latency, function to suppress lytic gene expression and enhance the survival of virus-infected neurons. The latent cell population comprises primary-order neurons infected directly from peripheral tissues and cells infected following further virus spread within the ganglion. In order to assess the role of LAT expression on latency establishment within first-order neurons, we infected ROSA26R reporter mice with Cre recombinase-expressing recombinant viruses harbouring deletion of the thymidine kinase lytic gene and/or the core LAT promoter. We found that LAT expression did not impact on latency establishment in viruses unable to replicate in neurons, and under these conditions, it was not required for the survival of neurons between 3 and 31 days post-infection.


A heterotopic pregnancy involving a caesarean section scar.
Heterotopic pregnancy refers to the presence of simultaneous pregnancies in two different implantation sites, generally one intra-uterine pregnancy and one extra-uterine pregnancy (usually tubal). This is a rare case of a heterotopic pregnancy involving concurrent intra-uterine pregnancy and caesarean section scar pregnancy (CSEP). CSEPs are at a high risk of bleeding and uterine rupture, carrying with them significant maternal morbidity.


Neovascularisation: has the angiogenic factor already been found?
Recently many soluble growth factors capable of influencing neovascularisation (angiogenesis) have been isolated and molecularly cloned. As such they are now available in a highly purified and active form. One or several of these already quite well known molecules may be of importance in the control of ocular neovascularisation. This article reviews what is presently known about growth factor control of neovascularisation with particular emphasis on both the eye and those factors that have already been molecularly cloned. In addition several recently reported inhibitors of neovascularisation are discussed. Such research is of particular interest to the ophthalmologist as knowledge gained in this area may allow for the use of both growth factors as well as growth factor inhibitors in the management of several ocular diseases involving neovascularisation.


Concomitant therapy: an outcome variable for musculoskeletal disorders? Part 2: total joint replacement in osteoarthritis trials.
Interest has grown in using the requirement of total joint replacement (TJR) as a "hard" outcome measure. Limitations exist, however, in the use of such an outcome, in particular the variability in the decision to perform surgery, length of surgical waiting lists, and sensitivity to change. This special interest group is exploring ways of retaining the clinical relevance of TJR but overcoming the problems--2 alternative outcomes are being considered: "time to physician's decision to recommend surgery" and "time to fulfilling criteria for total joint replacement."


The speed of feature-based attention: attentional advantage is slow, but selection is fast.
When paying attention to a feature (e.g., red), no attentional advantage is gained in perceiving items with this feature in very brief displays. Therefore, feature-based attention seems to be slow. In previous feature-based attention studies, attention has often been measured as the difference in performance in a secondary task. In our recent work on Boolean map theory (Huang & Pashler, 2007), we distinguished between 2 concepts that are often conflated with the term attention, namely the selection of information from stimulus and the following processing optimization (i.e., attentional advantage) of the selected stimulus. Attention, as examined in previous feature-based attention studies, only fits the definition of processing optimization, but does not fit the definition of selection of information. Therefore, it is open to question whether feature-based attention, when defined as selection, is fast or slow. In this study, I systematically measured the speed of feature-based attention in relation to both definitions. Attention was found to be slow (~100 ms) in terms of processing optimization (i.e., attentional advantage) but fast in terms of the selection of information (<50 ms). These results support the view that feature-based attention works by creating a spatial representation (i.e., a Boolean map; Huang & Pashler, 2007) for the stimulus of a feature and a processing optimization of the visual information residing in the region of this spatial representation.


Pharmaceutical cost management in an ambulatory setting using a risk adjustment tool.
Pharmaceutical expenditure is undergoing very high growth, and accounts for 30% of overall healthcare expenditure in Spain. In this paper we present a prediction model for primary health care pharmaceutical expenditure based on Clinical Risk Groups (CRG), a system that classifies individuals into mutually exclusive categories and assigns each person to a severity level if s/he has a chronic health condition. This model may be used to draw up budgets and control health spending. Descriptive study, cross-sectional. The study used a database of 4,700,000 population, with the following information: age, gender, assigned CRG group, chronic conditions and pharmaceutical expenditure. The predictive model for pharmaceutical expenditure was developed using CRG with 9 core groups and estimated by means of ordinary least squares (OLS). The weights obtained in the regression model were used to establish a case mix system to assign a prospective budget to health districts. The risk adjustment tool proved to have an acceptable level of prediction (R2 ≥ 0.55) to explain pharmaceutical expenditure. Significant differences were observed between the predictive budget using the model developed and real spending in some health districts. For evaluation of pharmaceutical spending of pediatricians, other models have to be established. The model is a valid tool to implement rational measures of cost containment in pharmaceutical expenditure, though it requires specific weights to adjust and forecast budgets.