High PARP-1 expression predicts poor survival in acute myeloid leukemia and PARP-1 inhibitor and SAHA-bendamustine hybrid inhibitor combination treatment synergistically enhances anti-tumor effects.
PARP-1 plays a critical role in DNA damage repair and contributes to progression of cancer. To explore the role of PARP-1 in acute myeloid leukemia (AML), we analyzed the expression of PARP-1 in AML and its relation to the clinical prognosis. Then, we investigated the efficacy and mechanism of PARP inhibitor BMN673 (Talazoparib) combined with NL101, a novel SAHA-bendamustine hybrid in vitro and in vivo. The expression of PARP-1 in 339 cytogenetically normal AML (CN-AML) cases was evaluated using RT-PCR. According to the expression of PARP-1, the clinical characteristics and prognosis of the patients were grouped and compared. The combination effects of BMN673 and NL101 were studied in AML cells and B-NSG mice xenograft model of MV4-11. We found patients in high PARP-1 expression group had higher levels of blast cells in bone marrow (P = .003) and white blood cells (WBC) in peripheral blood (P = .008), and were associated with a more frequent FLT3-ITD mutation (28.2% vs 17.3%, P = .031). The overall survival (OS) and event free survival (EFS) of the high expression group were significantly shorter than those in the low expression group (OS, P = .005 and EFS, P = .004). BMN673 combined with NL101 had a strong synergistic effect in treating AML. The combination significantly induced cell apoptosis and arrested cell cycle in G2/M phase. Mechanistically, BMN673 and NL101 combinatorial treatment promoted DNA damage. In vivo, the combination effectively delayed the development of AML and prolonged survival. High PARP-1 expression predicts poor survival in CN-AML patients. The synergistic effects of PARP inhibitor BMN673 in combination with SAHA-bendamustine hybrid, NL101, provide a new therapeutic strategy against AML. FUND: National Natural Science Foundation of China and Zhejiang Provincial Key Innovation Team.


Nanoscale Archimedean Tilings Formed by 3-Miktoarm Star Terpolymer Thin Films.
3-Miktoarm star terpolymer architecture (3µ-ABC), consisting of three dissimilar polymer chains, A, B, and C connected at a junction point, provides a unique opportunity in the design of complex nanoscale patterns such as Archimedean tilings that are not accessible from linear ABC terpolymers. In this work, the synthesis and the self-assembly of 3-miktoarm star terpolymers, namely, polystyrene-arm-poly(2-vinylpyridine)-arm-polyisoprene (3µ-SPI) into Archimedean tiling patterns is described. Several 3µ-SPI terpolymers are produced via a mid-functional PS-b-P2VP, synthesized by sequential anionic polymerization, using a 1,1-diphenylethylene bearing a tert-butyldimethylsilyl-protected hydroxyl functionality as a core molecule. PI arms with different sizes are then linked to the deprotected hydroxyl function of the core molecule via a Steglich esterification. Solvent-annealed 3µ-SPI thin films exhibit nanoscale prisms arranged into a (4.6.12) Archimedean tiling pattern. Depending on the size of the low etch-resistant PI arm and the solvent selected to promote the self-assembly of 3µ-SPI thin films, the voided columns occupy the square or decagonal sites of the (4.6.12) pattern. The use of this (4.6.12) tiling produced for the first time from self-assembled 3µ-ABC thin films can be a promising route to build 2D photonic crystals having complete photonic band gaps, where the light propagation is completely prohibited.


Phylodynamic analyses of Brazilian antigenic variants of infectious bursal disease virus.
Infectious bursal disease virus (IBDV) is a very important pathogen to poultry production and it is classified into three main groups: classical virulent (cvIBDV), very virulent (vvIBDV) and antigenic variants (avIBDV). This last group is composed by five different genetic lineages (recently classified in genogroups G2, G4, G5, G6, and G7) distributed in specific regions around the world. Brazil is one of the biggest poultry producers in the world and the present study aimed to investigate the evolutionary history of avIBDVs of the genogroup G4 in Brazil. A total of 5331 IBDV positive bursa samples, from different Brazilian poultry flocks, were genotyped in a period of ten years (2005 to 2014) and 1888 (35.42%) were identified as local avIBDVs. The highly variable region of the viral protein 2 (hvvp2) gene of 28 avIBDVs was sequenced and used in phylogenetic analyses and evaluation of local amino acid signatures. In addition, all complete and partial IBDV vp2 gene sequences, with local and year of collection information available on GenBank, were retrieved. Phylogenetic analyses were carried out based on a maximum likelihood method for the classification of genogroups occurring in Brazil. Based on a Maximum Likelihood (ML) phylogenetic tree, all Brazilian avIBDVs grouped into the genogroup 4. Bayesian phylodynamics analysis demonstrated the ancestor virus of this group was probably introduced in South America in 1968 (1960 to 1974, 95% HPD) and in Brazil in 1974 (1968 to 1977, 95% HPD) and the most likely source was East Europe (Hungary or Poland). All Brazilian avIBDV sequences, as well as the other genogroup 4 sequences, showed a specific pattern of amino acid: S222, T272, P289, I290, and F296. This report brings new insights about the IBDV epidemiology in Brazil and South America.


IBS: transformation of our governance.
In my Presidential Address at the 2010 International Biometric Conference in Florianopolis, I outlined the revised governance structure for the International Biometric Society (IBS). That structure was subsequently modified, with the final version being approved by the society membership in June 2012. The membership also approved the merger of the constitution with the bylaws, effectively dissolving the constitution (as it was no longer consistent with practice). From January 1, 2013, responsibility for the governance and leadership of the IBS rests with a 15-member Executive Board which will be supported by a larger Representative Council whose members are selected by and from each of the society's regions. The Representative Council is responsible for overseeing the determination of the Executive Board, providing advice on strategic and policy issues, and contributing to the operation of the IBS. This Council will be an effective conduit between the regions and the Executive Board, aided by the Chair attending all Board meetings.


Open-label, single-dose, parallel-group study in healthy volunteers to determine the drug-drug interaction potential between KAE609 (cipargamin) and piperaquine.
KAE609 represents a new class of potent, fast-acting, schizonticidal antimalarials. This study investigated the safety and pharmacokinetics of KAE609 in combination with the long-acting antimalarial piperaquine (PPQ) in healthy volunteers. A two-way pharmacokinetic interaction was hypothesized for KAE609 and PPQ, as both drugs are CYP3A4 substrates and inhibitors. The potential for both agents to affect the QT interval was also assessed. This was an open-label, parallel-group, single-dose study with healthy volunteers. Subjects were randomized to four parallel dosing arms with five cohorts (2:2:2:2:1), receiving 75 mg KAE609 plus 320 mg PPQ, 25 mg KAE609 plus 1,280 mg PPQ, 25 mg KAE609 alone, 320 mg PPQ alone, or 1,280 mg PPQ alone. Triplicate electrocardiograms were performed over the first 24 h after dosing, with single electrocardiograms at other time points. Routine safety (up to 89 days) and pharmacokinetic (up to 61 days) assessments were performed. Of the 110 subjects recruited, 99 completed the study. Coadministration of PPQ had no overall effect on exposure to KAE609, although 1,280 mg PPQ decreased the KAE609 maximum concentration (Cmax) by 17%. The group that received 25 mg KAE609 plus 1,280 mg PPQ showed a 32% increase in the PPQ area under the concentration-time curve from 0 to infinity (AUCinf), while the group that received 75 mg KAE609 plus 320 mg PPQ showed a 14% reduction. Mean changes from baseline in the QT interval corrected by Fridericia's method (QTcF) and the QT interval corrected by Bazett's method (QTcB) with PPQ were consistent with its known effects. PPQ but not KAE609 exposure correlated with corrected QT interval (QTc) increases, and KAE609 did not affect the PPQ exposure-QTc relationship. The QTcF effect for PPQ (least-squares estimate of the difference in mean maximal changes from baseline of 7.47 ms [90% confidence interval, 3.55 to 11.4 ms]) was consistent with the criteria for a positive thorough QT study. No subject had QTcF or QTcB values of >500 ms. Both drugs given alone or in combination were well tolerated, with no deaths, serious adverse events (AEs), or severe AEs reported. Most AEs were mild; upper respiratory tract infections, headache, diarrhea, and oropharyngeal pain were most common. PPQ and KAE609 coadministration had no relevant effect on exposure to either agent, and KAE609 did not affect or potentiate the known effects of PPQ on cardiac conduction.


Identification of amino acid residues of the T-cell epitope of Mycobacterium tuberculosis alpha antigen critical for Vbeta11(+) Th1 cells.
Stimulation of Mycobacterium tuberculosis-primed lymph node cells from C57BL/6 mice with alpha antigen (also known as antigen 85B and MPT59) induced cell proliferation, production of interleukin 2 and gamma interferon, and expansion of Vbeta11(+) CD4(+) T cells in conjunction with antigen-presenting cells in an I-A(b)-restricted manner. Using a series of 15-amino-acid peptides that overlapped each other by 5 amino acids and spanned the mature alpha antigen, we identified the antigenic epitope for alpha antigen-specific Vbeta11(+) Th1 cells. That peptide (peptide-25), which corresponds to amino acid residues 240 to 254 of alpha antigen, contains a motif that is conserved in I-A(b) and requires processing by antigen-presenting cells. Using peptide-25-reactive Vbeta11(+) T-cell clones and substituted peptide-25 mutants, we determined which amino acid residues within peptide-25 were critical for T-cell receptor (TCR) recognition. Our results showed that the amino acid residues at positions 245, 246, 248, 250, and 251 are important for recognition of TCRVbeta11 and that residues at positions 244, 247, 249, and 252 are I-A(b) contact residues. We also observed that active immunization of C57BL/6 mice with peptide-25 can lead to decreased bacterial load in the lungs of M. tuberculosis H37Rv-infected mice. These results should provide us with a useful tool for delineating the regulation of Vbeta11(+) Th1-cell development during M. tuberculosis infection and for developing a vaccine inducing a Th1-dominant immune response.


Experimental and clinical applications and outcomes of using different forms of suction in retrograde intrarenal surgery. Results from a systematic review.
To identify the latest advances in suction devices and evaluate their effect in Retrograde intrarenal surgery (RIRS) and ureteroscopy for stones. A systematic literature search was performed on 4th January 2023 using Scopus, PubMed, and EMBASE. Only English papers were included; both pediatric and adult studies were accepted. Duplicate studies, case reports, letters to the editor, and meeting abstracts were excluded. Twenty-one papers were selected. Several methods have been proposed for suction use in RIRS, such as through the ureteral access sheath or directly to the scope. Artificial intelligence can also regulate this system, monitoring pressure and perfusion flow values. All the proposed techniques showed satisfactory perioperative results for operative time, stone-free rate (SFR), and residual fragments. Moreover, the reduction of intrarenal pressure (induced by aspiration) was also associated with a lower infection rate. Even the studies that considered kidney stones with a diameter of 20 mm or higher reported higher SFR and reduced postoperative complications. However, the lack of well-defined settings for suction pressure and fluid flow prevents the standardization of the procedure. Aspiration device in the surgical treatment of urinary stones favours a higher SFR, reducing infectious complications, as supported by the included studies. RIRS with a suction system provided to be a natural successor to the traditional technique, regulating intrarenal pressure and aspirating fine dust.


Idiopathic intracranial hypertension in the Middle East: A growing concern.
Idiopathic Intracranial Hypertension (IIH) is a disorder of increased intracranial pressure without any identifiable etiology. It is defined by elevated intracranial pressure (ICP) with normal neuroimaging and normal cerebrospinal fluid (CSF) contents. IIH typically affects young obese women and produces symptoms and signs related to high ICP. Headache and blurred vision are the most common symptoms, and papilledema is the major clinical sign. In this review we examine the epidemiology and demographic features of IIH in Middle Eastern countries and compare and contrast them with the published IIH literature from Western countries. The incidence of IIH in several Middle East countries has been estimated at 2.02-2.2/100,000 in the general population, which is higher than the Western rate. Obesity is a major risk factor globally and it is associated with an increased risk of severe vision loss due to IIH. There has been an increase in obesity prevalence in the Middle East countries mainly affecting the Gulf Council Countries (GCC), which parallels increased industrial development. This rise may be contributing to the increasing incidence of IIH in these countries. Other risk factors may also be contributing to IIH in Middle East countries and the differences and similarities to Western IIH merit further study.


Regulation of bovine rod outer segment membrane guanylate cyclase by ATP, phosphodiesterase and metal ions.
In vertebrate retina, rod outer segment is the site of visual transduction. The inward cationic current in the dark-adapted outer segment is regulated by cyclic GMP. A light flash on the outer segment activates a cyclic GMP phosphodiesterase resulting in rapid hydrolysis of the cyclic nucleotide which in turn causes a decrease in the dark current. Restoration of the dark current requires inactivation of the phosphodiesterase and synthesis of cyclic GMP. The latter is accomplished by the enzyme guanylate cyclase which catalyzes the formation of cyclic GMP from GTP. Therefore, factors regulating the cyclase activity play a critical role in visual transduction. But regulation of the cyclase by some of these factors--phosphodiesterase, ATP, the soluble proteins and metal cofactors (Mg and Mn)--is controversial. The availability of different types of cyclase preparations, dark-adapted rod outer segments with fully inhibited phosphodiesterase activity, partially purified cyclase without PDE contamination, cloned rod outer segment cyclase free of other rod outer segment proteins, permitted us to address these controversial issues. The results show that ATP inhibits the basal cyclase activity but enhances the stimulation of the enzyme by soluble activator, that cyclase can be activated in the dark at low calcium concentrations under conditions where phosphodiesterase activity is fully suppressed, and that greater activity is observed with manganese as cofactor than magnesium. These results provide a better understanding of the controls on cyclase activity in rod outer segments and suggest how regulation of this cyclase by ATP differs from that of other known membrane guanylate cyclases.


Examining the reliability and factor structure of the Malay version of the Recovery Self-Assessment in a sample of individuals with schizophrenia.
Without a reliable and valid tool, it is neither possible to gauge recovery orientation of persons with serious mental illness nor to evaluate whether existing mental health system encourages recovery in both its policies and current practice in Malaysia. Therefore, the aim of this study was to examine the reliability and factor structure of the Malay version of the Recovery Self-Assessment (RSA-PIRV-M) in a psychiatric sample. Psychiatric patients were eligible for recruitment to the study, when they fulfilled the following criteria: a diagnosis of schizophrenia, were aged between 18- and 65-years-old, and were able to give consent themselves. We recruited 118 individuals with schizophrenia. They were selected via systematic random sampling technique. All RSA-PIRV-M items were derived from the parent scale. These items were translated based on established procedures. The reliability estimates of the RSA-PIRV-M were based on Cronbach's alpha. We performed confirmatory factor analyses to examine the factor structure of the RSA-PIRV-M. All Cronbach's alphas for the RSA-PIRV-M subscales were at least .70. With respect to the factor structure of the RSA-PIRV-M, our structural equation modeling findings suggest a five-factor model encompassing life goals, involvement, diversity of treatment options, choice, and individually-tailored services. The interpretation of our findings is limited by small sample size, unique sample characteristics, and lack of further evidence for convergent validity. Notwithstanding these limitations, the RSA-PIRV-M is a promising tool in bridging gaps between our knowledge on recovery orientation and existing mental health service provision in Malaysia. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Hip and ankle range of motion and hip muscle strength in young female ballet dancers and controls.
To compare the hip and ankle range of motion and hip muscle strength in 8-11 year old novice female ballet dancers and controls. Subjects were 77 dancers and 49 controls (mean (SD) age 9.6 (0.8) and 9.6 (0.7) years respectively). Supine right active hip external rotation (ER) and internal rotation (IR) were measured using an inclinometer. A turnout protractor was used to assess standing active turnout range. The measure of ER achieved from below the hip during turnout (non-hip ER) was calculated by subtracting hip ER range from turnout range, and hip ER:IR was derived by dividing ER range by IR range. Range of right weight bearing ankle dorsiflexion was measured in a standing lunge using two methods: the distance from the foot to the wall (in centimetres) and the angle of the shank to the vertical via an inclinometer (in degrees). Right calf muscle range was measured in weight bearing using an inclinometer. A manual muscle tester was used to assess right isometric hip flexor, internal rotator, external rotator, abductor, and adductor strength. Dancers had less ER (p<0.05) and IR (p<0.01) range than controls but greater ER:IR (p<0.01). Although there was no difference in turnout between groups, the dancers had greater non-hip ER. Dancers had greater range of ankle dorsiflexion than controls, measured in both centimetres (p<0.01) and degrees (p<0.05), but similar calf muscle range. After controlling for body weight, controls had stronger hip muscles than dancers except for hip abductor strength which was similar. Regression analyses disclosed a moderate relation between turnout and hip ER (r = 0.40). There were no significant correlations between range of motion and training years and weekly training hours. Longitudinal follow up will assist in determining whether or not hip and ankle range in young dancers is genetically fixed and unable to be improved with further balletic training.


Circular permutation of Bacillus circulans xylanase: a kinetic and structural study.
The 20 kDa Bacillus circulans Bcx is a well-studied endoxylanase with a beta-jellyroll fold that places its N- and C-termini in salt bridge contact. Initial experiments verified that Bcx could be circularly permuted by PCR methods to introduce new termini in loop regions while linking its native termini directly or via one or two glycines. Subsequently, a library of circular permutants, generated by random DNase cleavage of the circularized Bcx gene, was screened for xylanase activity on xylan in Congo Red-stained agar. Analysis of 35 unique active circular permutants revealed that, while many of the new termini were introduced in external loops as anticipated, a surprising number were also located within beta-strands. Furthermore, several permutations placed key catalytic residues at or near the new termini with minimal deleterious effects on activity and, in one case, a 4-fold increase. The structure of one permutant was determined by X-ray crystallography, whereas three others were probed by NMR spectroscopy. These studies revealed that the overall conformation of Bcx changed very little in response to circular permutation, with effects largely being limited to increased local mobility near the new and the linked old termini and to a decrease in global stability against thermal denaturation. This library of circularly permuted xylanases provides an excellent set of new start points for directed evolution of this commercially important enzyme, as well as valuable constructs for intein-mediated replacement of key catalytic residues with unnatural analogues. Such approaches should permit new insights into the mechanism of enzymatic glycoside hydrolysis.


hCG: Biological Functions and Clinical Applications.
Human chorionic gonadotropin (hCG) is produced primarily by differentiated syncytiotrophoblasts, and represents a key embryonic signal that is essential for the maintenance of pregnancy. hCG can activate various signaling cascades including mothers against decapentaplegic homolog 2 (Smad2), protein kinase C (PKC), and/or protein kinase A (PKA) in several cells types by binding to luteinizing hormone/chorionic gonadotropin receptor (LHCGR) or potentially by direct/indirect interaction with transforming growth factor beta receptor (TGFβR). The molecule displays specialized roles in promoting angiogenesis in the uterine endothelium, maintaining myometrial quiescence, as well as fostering immunomodulation at the maternal-fetal interface. It is a member of the glycoprotein hormone family that includes luteinizing hormone (LH), thyroid-stimulating hormone (TSH), and follicle-stimulating hormone (FSH). The α-subunit of hCG displays homologies with TSH, LH, and FSH, whereas the β subunit is 80-85% homologous to LH. The hCG molecule is produced by a variety of organs, exists in various forms, exerts vital biological functions, and has various clinical roles ranging from diagnosis and monitoring of pregnancy and pregnancy-related disorders to cancer surveillance. This review presents a detailed examination of hCG and its various clinical applications.


Functional characterization of AF-04, an afucosylated anti-MARV GP antibody.
Marburg virus (MARV), one member of the Filoviridae family, cause sporadic outbreaks of hemorrhagic fever with high mortality rates. No countermeasures are currently available for the prevention or treatment of MARV infection. Monoclonal antibodies (mAbs) are promising candidates to display high neutralizing activity against MARV infection in vitro and in vivo. Recently, growing evidence has shown that immune effector function including antibody-dependent cell-mediated cytotoxicity (ADCC) is also required for in vivo efficacy of a panel of antibodies. Glyco-engineered methods are widely utilized to augment ADCC function of mAbs. In this study, we generated a fucose-knockout MARV GP-specific mAb named AF-04 and showed that afucosylation dramatically increased its binding affinity to polymorphic FcγRIIIa (F176/V176) compared with the parental AF-03. Accordingly, AF-04-mediated NK cell activation and NFAT expression downstream of FcγRIIIa in effector cells were also augmented. In conclusion, this work demonstrates that AF-04 represents a novel avenue for the treatment of MARV-caused disease.


Influence of Risk Perception on Task and Contextual Performance: A Case of Work-Related Musculoskeletal Disorders in Nurses.
Assessing the risk perception of workers can be very informative in predicting their behavioral outcomes, including task and contextual performance. Yet, research to assess the effect of risk perception on task performance and contextual performance remains scarce. Thus, this study explored the effect of risk perception of work-related musculoskeletal disorders on task and contextual performance in nurses. This study further examined safety behavior as a mediator of these relationships. Using structural equation modeling, the researchers examined these relationships by employing a cross-sectional survey with a quantitative approach. The data was collected via questionnaires from 382 nurses who work in three major hospitals in Accra Metropolis, Ghana. The results showed that nurses' risk perception positively influenced their task and contextual performance, and that safety behavior partially mediated the effects of risk perception on task and contextual performance. This study offers a theoretical framework and empirical evidence for the concept of risk perception and its association with safety behavior, task, and contextual performance. To the best of our knowledge, this is the first study to assess the relationships that exist among these variables. Thus, future studies are needed to verify the causality of the relationships.


Drug distribution in spinal cord during administration with spinal loop dialysis probes in anaesthetized rats.
The present investigation aimed to study two methodological concerns of an experimental model, where a spinal loop dialysis probe is used for administration of substances to the spinal cord and sampling of neurotransmitters by microdialysis from the same area of anaesthetized rats. [(3)H]Epibatidine in concentrations of 1, 10 and 100 nM was dissolved in Ringer's solution and administered through the dialysis membrane into the dorsal region of the cervical spinal cord. First, the outflow of [(3)H]epibatidine from the probe into the spinal cord was examined with respect to different concentrations and changes over time. Then, the distribution of the different [(3)H]epibatidine concentrations along the spinal cord was studied. It was found that the percentage of [(3)H]epibatidine entering the spinal cord did not differ between different administered concentrations after a stabilization period of 60 min. The administered [(3)H]epibatidine was found to be distributed to the area closest to the dialysis probe and not dispersed along the spinal cord, and the distribution was equal for all concentrations. The data presented in this investigation provide information, which is important for interpretation of data from intraspinal administration of substances through the spinal loop dialysis probe.


Localization and detection of breast cancer tumors with digital image elasto-tomography.
Digital Image Elasto-Tomography (DIET) is a novel elastic contrast based breast imaging method using time-harmonic motion data obtained from a calibrated array of high resolution digital cameras scanning the tissue surface. The method is currently undergoing initial clinical testing and preliminary results in cases of malignant breast tumors are now available. The method has proved capable of detecting and localizing the stiff lesions within the heterogeneous tissue structure of the beast through the use of an evolution based optimization algorithm implemented in linear finite elements. The method has also proved successful at detecting both inclusion and non-inclusion cases in specially constructed tissue mimicking silicon phantoms.


Effect of prolactin on testicular lipid metabolism in pubertal rats.
Prolactin administration for 7 day to pubertal rats resulted in marked depletion of total lipids, phospholipids and particularly phosphatidyl choline and phosphatidyl ethanolamine. There was a concomitant increase in total cholesterol and cholesterol ester with a fall in free cholesterol. The increase in total cholesterol in the testis of prolactin-treated animals appears not to be due to its increased synthesis, as the hormonal treatment had no significant effect on HMG-Co A reductase, the rate limiting enzyme of sterol synthesis. Prolactin also had no significant effect on the enzymes glucose-6-phosphate dehydrogenase, malic- and isocitrate dehydrogenases reported to generate NADPH required for active steroidogenesis. Thus, it appears that the fall observed in testicular phospholipids and free cholesterol with concurrent increases in total cholesterol and cholesterol ester after prolactin administration is not due to prolactin's effect directly on testicular lipid metabolism in pubertal rats.


Nomenclatural changes in Centroptella Braasch & Soldán, 1980 (Ephemeroptera, Baetidae).
The genus Centroptella Braasch & Soldán, 1980 is accepted here in a wide sense, i.e., including Chopralla Waltz & McCafferty, 1987. This genus concept is similar to the concept of the genus Bungona Harker, 1957 proposed by Salles et al. (2016), but with the generic name Centroptella instead of Bungona. The type species of Bungona, B. narilla Harker, 1957, has an unknown systematic position; the neotype designation proposed by Suter and Pearson (2001) is invalid, being inconsistent with the International Code of Zoological Nomenclature; the species name B. narilla and the generic name Bungona are nomina dubia, so the name Centroptella is the senior name for the genus under consideration. The generic names Chopralla and Crassolus Salles, Gattolliat & Sartori, 2016 both are junior synonyms of Centroptella (syn. nov.). The subgenera Bungona, Centroptella and Chopralla proposed by Salles et al. (2016) are unnatural. The following new combinations are proposed: Centroptella bintang (Marle, Salles & Gattolliat, 2016) comb. nov., Centroptella bifida (Shi & Tong, 2019) comb. nov., Centroptella fusina (Tong & Dudgeon, 2003) comb. nov., Centroptella fustipalpus (Lugo-Ortiz & McCafferty, 1998) comb. nov., Centroptella illiesi (Lugo-Ortiz & McCafferty, 1998) comb. nov., Centroptella inzingae (Crass, 1947) comb. nov., Centroptella papilionodes (Marle, Salles & Gattolliat, 2016) comb. nov., Centroptella pontica (Sroka, Godunko & Gattolliat, in Sroka et al. 2019) comb. nov., Centroptella ovata (Shi & Tong, 2019) comb. nov., Centroptella quadrata (Shi & Tong, 2019) comb. nov. and Centroptella saxophila (Agnew, 1961) comb. nov. The two Australian species, C. fustipalpus and C. illiesi, differ from each other in the shape of tergalii; corrections to the original description of C. fustipalpus are given based on re-examination of the holotype and paratypes; details of larval structures of C. illiesi are figured. Corrections to the former descriptions of the South African species C. inzingae and C. saxophila are given. Examination of type material led to the discovery that the original description of the Oriental species Centroptella liebenauae Soldán, Braasch & Muu, 1987 was based on two different species: the descriptions of imago and subimago belong to Centroptella longisetosa Braasch & Soldán, 1980 (the type species of Centroptella), and the description of larva belongs to a different species, which we describe here as Centroptella ingridae sp. nov. The holotype of C. liebenauae, a larva, should be considered lost; based on the date of collection, it belonged to C. longisetosa; a set of larval exuviae with the same collecting data as the holotype, is designated as the neotype of C. liebenauae, and a new synonymy is established: C. longisetosa = C. liebenauae syn. nov. The larvae originally assigned to C. liebenauae are placed to a new species Centroptella ingridae sp. nov. belonging to the inzingae-ingridae species group; all stages of development of this species are described based on male and female imagines reared from larvae in Thailand and on the misidentified paratypes of C. liebenauae from Vietnam. Centroptella longisetosa is redescribed based on the single paratype from China, the neotype and paratypes of C. liebenauae from Vietnam, and additional material from India. Additional data on the holotype of Centroptella colorata Soldán, Braasch & Muu, 1987 are given.


Pulmonary sequestration associated with tuberculosis, aspergillosis and pseudomycosis.
Three cases of pulmonary sequestrations are reported, one associated with tuberculosis, another with aspergillosis and a third cases associated with pseudomycosis (bothryomycosis). All the patients had a localized infection. They were treated surgically and had an uneventful recovery from their disease.


In vitro evaluation of lipids adsorbed on silicone hydrogel contact lenses using a new gas chromatography/mass spectrometry analytical method.
To establish a new analytical method using gas chromatography/mass spectrometry (GC/MS) for evaluating the lipids adsorbed on silicone hydrogel (SH) contact lenses: cholesterol, cholesterol ester, wax, and squalene. A novel GC/MS method was developed and validated for the repeatability, specificity, linearity, detection and quantitation limits, and percentage recovery. The lipids in an artificial tear solution were adsorbed on 5 SH lenses (asmofilcon A, balafilcon A, galyfilcon A, lotrafilcon A, and lotrafilcon B) and 1 conventional hydrogel lens (etafilcon A) in vitro. The lipids adsorbed were then extracted and analyzed by the GC/MS method. Repeatability of this analytical method was less than 2.2% deviation for all test lipids; however, the analytes were completely discriminated with sharp peak shapes, and identified specifically. The correlation coefficient showing linearity was at least 0.991 under 50 microg/mL of lipid concentration. Detection and quantitation limits were statistically 0.5 to 0.8 microg/mL and 1.4 to 2.5 microg/mL, respectively, for all analytes. Percentage recovery was estimated as approximately 80% for 3 microg/lens, 90% for 5 microg/lens, and almost 100% for larger amounts of lipids. Quantitatively, the lipids absorbed on contact lenses were lotrafilcon A = (near equal) lotrafilcon B = (near equal) etafilcon A < asmofilcon A < galyfilcon A = (near equal) balafilcon A with the value of 0.4 to 7.6 microg/lens. Lipid adsorption on SH lenses varied depending on the lipid components and lens surface properties. The GC/MS method established in this study is excellent for the repeatability, specificity, linearity, detection and quantitation limits, and percentage recovery, and provides a novel highly sensitive and useful tool for evaluating lipids adsorbed on SH lenses.


Direction of force generated by the inner row of dynein arms on flagellar microtubules.
Our goal was to determine the direction of force generation of the inner dynein arms in flagellar axonemes. We developed an efficient means of extracting the outer row of dynein arms in demembranated sperm tail axonemes, leaving the inner row of dynein arms structurally and functionally intact. Sperm tail axonemes depleted of outer arms beat at half the beat frequency of sperm tails with intact arms over a wide range of ATP concentrations. The isolated, outer arm-depleted axonemes were induced to undergo microtubule sliding in the presence of ATP and trypsin. Electron microscopic analysis of the relative direction of microtubule sliding (see Sale, W. S. and P. Satir, 1977, Proc. Natl. Acad. Sci. USA, 74:2045-2049) revealed that the doublet microtubule with the row of inner dynein arms, doublet N, always moved by sliding toward the proximal end of the axoneme relative to doublet N + 1. Therefore, the inner arms generate force such that doublet N pushes doublet N + 1 tipward. This is the same direction of microtubule sliding induced by ATP and trypsin in axonemes having both inner and outer dynein arms. The implications of this result for the mechanism of ciliary bending and utility in functional definition of cytoplasmic dyneins are discussed.


Significance of diagnostic parameters in [(13)c]octanoic Acid gastric emptying breath tests*.
Abstract Two novel characteristic parameters, the latency time (t (lat)) and the ascension time (t (asc)), are proposed for evaluation of non-invasive [(13)C]octanoic acid breath tests for assessment of the gastric emptying of solids. In breath tests performed in control subjects (n = 30) and diabetic patients (n = 100), the usefulness of these parameters was compared to conventional parameters, i.e., gastric half emptying-time t (1/2,b )) and lag phase (t (lag,b )). The proposed parameters were only loosely correlated (controls, r = 0.199; diabetics, 0.616). A strong correlation was found between the conventional parameters (controls, r = 0.891; diabetics, r = 0.962). Based on the conventional method, 36 patients were suspicious of delayed gastric emptying including 24 patients which exhibited a simultaneous delay in both parameters. Using the new parameters, a total of 46 patients were suspicious of delayed gastric emptying with 15 and 20 having isolated delay in t (lat) and t (asc), respectively. We conclude that the novel parameters may be more appropriate for examination of the different phases of gastric emptying and for evaluation of gastric emptying disturbances in diabetic patients than the parameters conventionally used for this purpose.


A label-free selected reaction monitoring workflow identifies a subset of pregnancy specific glycoproteins as potential predictive markers of early-onset pre-eclampsia.
Pre-eclampsia (PE) is a serious complication of pregnancy with potentially life threatening consequences for both mother and baby. Presently there is no test with the required performance to predict which healthy first-time mothers will go on to develop PE. The high specificity, sensitivity, and multiplexed nature of selected reaction monitoring holds great potential as a tool for the verification and validation of putative candidate biomarkersfor disease states. Realization of this potential involves establishing a high throughput, cost effective, reproducible sample preparation workflow. We have developed a semi-automated HPLC-based sample preparation workflow before a label-free selected reaction monitoring approach. This workflow has been applied to the search for novel predictive biomarkers for PE. To discover novel candidate biomarkers for PE, we used isobaric tagging to identify several potential biomarker proteins in plasma obtained at 15 weeks gestation from nulliparous women who later developed PE compared with pregnant women who remained healthy. Such a study generates a number of "candidate" biomarkers that require further testing in larger patient cohorts. As proof-of-principle, two of these proteins were taken forward for verification in a 100 women (58 PE, 42 controls) using label-free SRM. We obtained reproducible protein quantitation across the 100 samples and demonstrated significant changes in protein levels, even with as little as 20% change in protein concentration. The SRM data correlated with a commercial ELISA, suggesting that this is a robust workflow suitable for rapid, affordable, label-free verification of which candidate biomarkers should be taken forward for thorough investigation. A subset of pregnancy-specific glycoproteins (PSGs) had value as novel predictive markers for PE.


End-of-life care pathways for improving outcomes in caring for the dying.
This is an updated version of a Cochrane review first published in Issue 1, 2010 of The Cochrane Library. In many clinical areas, integrated care pathways are utilised as structured multidisciplinary care plans that detail essential steps in caring for patients with specific clinical problems. In particular, care pathways for the dying have been developed as a model to improve care of patients who are in the last days of life. The care pathways were designed with an aim of ensuring that the most appropriate management occurs at the most appropriate time and that it is provided by the most appropriate health professional. There have been sustained concerns about the safety of implementing end-of-life care pathways, particularly in the UK. Therefore, there is a significant need for clinicians and policy makers to be informed about the effects of end-of-life care pathways with a systematic review. To assess the effects of end-of-life care pathways, compared with usual care (no pathway) or with care guided by another end-of-life care pathway across all healthcare settings (e.g. hospitals, residential aged care facilities, community).In particular, we aimed to assess the effects on symptom severity and quality of life of people who are dying; those related to the care such as families, carers and health professionals; or a combination of these. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (Issue 6, 2013), MEDLINE, EMBASE, PsycINFO, CINAHL, review articles and reference lists of relevant articles. We conducted the original search in September 2009, and the updated search in June 2013. All randomised controlled trials (RCTs), quasi-randomised trial or high-quality controlled before-and-after studies comparing use versus non-use of an end-of-life care pathway in caring for the dying. Two review authors assessed the results of the searches against the predetermined criteria for inclusion. The original review identified 920 titles. The updated search found 2042 potentially relevant titles (including the original 920), but no additional studies met criteria for inclusion in the review update. With sustained concerns about the safety of the pathway implementation and the lack of available evidence on important patient and relative outcomes, recommendations for the use of end-of-life pathways in caring for the dying cannot be made. Since the last version of this review, no new studies met criteria for inclusion in the review update. With recently documented concerns related to the potential adverse effects associated with Liverpool Care Pathway (the most commonly used end-of-life care pathway), we do not recommend decision making based on indirect or low-quality evidence. All health services using end-of-life care pathways are encouraged to have their use of the pathway, to date, independently audited. Any subsequent use should be based on carefully documented evaluations. Large RCTs or other well-designed controlled studies are urgently required for the evaluation of the use of end-of-life care pathways in caring for dying people in various clinical settings. In future studies, outcome measures should include benefits or harms concerning the outcomes of interest in this review in relation to patients, families, carers and health professionals.


Indonesian couples' pregnancy ambivalence and contraceptive use.
Most studies on pregnancy ambivalence are based on data from women and depend on the women's perceptions to measure their partner's pregnancy intentions. Because these perceptions may not be accurate, data collected directly from men are needed to understand the role of couple dynamics in fertility behavior. Matched couple data from the 2002-2003 Indonesia Demographic and Health Survey were used to examine contraceptive use, fertility desires and attitudes about becoming pregnant in the next few weeks-whether it would be a big problem, a small problem or no problem. Concordance between partners on these issues was evaluated. Inconsistent fertility desires and responses to the problem question are used to define ambivalence within couples. Multivariate logistic regression analyses were used to assess whether couples' pregnancy ambivalence was associated with contraceptive use. Seventy-one percent of husbands and 54% of wives reported that a pregnancy in the next few weeks would be "no problem"; couples' concordance on this question was 64% among contraceptive users and 61% among nonusers. In the multivariate analysis, couples who were discordant on the issue of a pregnancy in the near future had 26% lower odds of using contraceptives than couples in which both partners agreed a pregnancy would be a big or small problem. Contraceptive use was also less likely for couples in which one partner wanted to delay or stop childbearing and the other wanted more children or was undecided (odds ratio, 0.4). Husbands and wives influence each other's fertility attitudes and family planning use. Both husbands' and wives' pregnancy attitudes should be taken into account at the time of screening and method selection.


Differential regulation of drug transporter expression by all-trans retinoic acid in hepatoma HepaRG cells and human hepatocytes.
All-trans retinoic acid (atRA) is the active form of vitamin A, known to activate retinoid receptors, especially the heterodimer retinoid X receptor (RXR):retinoic acid receptor (RAR) that otherwise may play a role in regulation of some drug transporters. The present study was designed to characterize the nature of human hepatic transporters that may be targeted by atRA and the heterodimer RXR:RAR. Exposure of human hepatoma HepaRG cells and primary human hepatocytes to 5 μM atRA down-regulated mRNA levels of various sinusoidal solute carrier (SLC) influx transporters, including organic anion transporting polypeptide (OATP) 2B1, OATP1B1, organic cation transporter (OCT) 1 and organic anion transporter (OAT) 2, and induced those of the canalicular breast cancer resistance protein (BCRP). The retinoid concomitantly reduced protein expression of OATP2B1 and OATP1B1 and activity of OATPs and OCT1 and induced BCRP protein expression in HepaRG cells. Some transporters such as OATP1B3 and the bile salt export pump (BSEP) were however down-regulated by atRA in primary human hepatocytes, but induced in HepaRG cells, thus pointing out discrepancies between these two liver cell models in terms of detoxifying protein regulation. atRA-mediated repressions of OATP2B1, OATP1B1, OAT2 and OCT1 mRNA expression were finally shown to be counteracted by knocking-down expression of RARα and RXRα through siRNA transfection in HepaRG cells. atRA thus differentially regulated human hepatic drug transporters, mainly in a RXR:RAR-dependent manner, therefore establishing retinoids and retinoid receptors as modulators of liver drug transporter expression.


[Hirsutism. Diagnosis and therapy].
Increased hair growth represents a severe problem for many women. In rare cases an adrenal or ovarian cause can be detected which can be treated specifically. Drug-induced hirsutism and other rare syndromes must be excluded. Idiopathic hirsutism is to be assumed if no clear-cut evidence for ovarian or adrenal disease can be found. Idiopathic hirsutism can be treated symptomatically with anti-androgens. However, after anti-androgen withdrawal hirsutism often recurs, necessitating additional cosmetic therapy.


Animal models for evaluation of oral delivery of biopharmaceuticals.
Biopharmaceuticals are increasingly important for patients and the pharmaceutical industry due to their ability to treat and, in some cases, even cure chronic and potentially life-threatening diseases. Most biopharmaceuticals are administered by injection, but intensive focus on development of systems for oral delivery of biopharmaceuticals may result in new treatment modalities to increase the patient compliance and reduce product cost. In the preclinical development phase, use of experimental animal models is essential for evaluation of new formulation designs. In general, the limited oral bioavailability of biopharmaceuticals, of just a few percent, is expected, and therefore, the animal models and the experimental settings must be chosen with utmost care. More knowledge and focus on this topic is highly needed, despite experience from the numerous studies evaluating animal models for oral drug delivery of small molecule drugs. This review highlights and discusses pros and cons of the most currently used animal models and settings. Additionally, it also looks into the influence of anesthetics and sampling methods for evaluation of drug delivery systems for oral delivery of biopharmaceuticals primarily with examples on insulin.


Thinking in three's: changing surgical patient safety practices in the complex modern operating room.
The three surgical patient safety events, wrong site surgery, retained surgical items (RSI) and surgical fires are rare occurrences and thus their effects on the complex modern operating room (OR) are difficult to study. The likelihood of occurrence and the magnitude of risk for each of these surgical safety events are undefined. Many providers may never have a personal experience with one of these events and training and education on these topics are sparse. These circumstances lead to faulty thinking that a provider won't ever have an event or if one does occur the provider will intuitively know what to do. Surgeons are not preoccupied with failure and tend to usually consider good outcomes, which leads them to ignore or diminish the importance of implementing and following simple safety practices. These circumstances contribute to the persistent low level occurrence of these three events and to the difficulty in generating sufficient interest to resource solutions. Individual facilities rarely have the time or talent to understand these events and develop lasting solutions. More often than not, even the most well meaning internal review results in a new line to a policy and some rigorous enforcement mandate. This approach routinely fails and is another reason why these problems are so persistent. Vigilance actions alone have been unsuccessful so hospitals now have to take a systematic approach to implementing safer processes and providing the resources for surgeons and other stakeholders to optimize the OR environment. This article discusses standardized processes of care for mitigation of injury or outright prevention of wrong site surgery, RSI and surgical fires in an action-oriented framework illustrating the strategic elements important in each event and focusing on the responsibilities for each of the three major OR agents-anesthesiologists, surgeons and nurses. A Surgical Patient Safety Checklist is discussed that incorporates the necessary elements to bring these team members together and influence the emergence of a safer OR.


Deletion of Thioredoxin-Interacting Protein (TXNIP) Abrogates High Fat Diet-induced Retinal Leukostasis, Barrier Dysfunction and Microvascular Degeneration in a Mouse Obesity Model.
We have shown that a high fat diet (HFD) induces the activation of retinal NOD-like receptor protein (NLRP3)-inflammasome that is associated with enhanced expression and interaction with thioredoxin-interacting protein (TXNIP). Here, the specific contribution of TXNIP and the impact of HFD on retinal leukostasis, barrier dysfunction and microvascular degeneration were investigated. Wild-type (WT) and TXNIP knockout (TKO) mice were fed with normal diet or 60% HFD for 8-18 weeks. TXNIP was overexpressed or silenced in human retinal endothelial cells (REC). At 8 weeks, HFD significantly induced retinal leukostasis and breakdown of the blood-retina barrier in WT mice, but not in TKO mice. In parallel, HFD also induced retinal expression of adhesion molecules and cleaved IL-1β in WT mice, which were also abrogated in TKO mice. In culture, TXNIP overexpression induced NLRP3, IL-1b, and adhesion molecules expression, while TXNIP silencing inhibited them. Blocking the IL-1β receptor significantly suppressed TXNIP-induced expression of NLRP3-inflammasome and adhesion molecules in HREC. Ex-vivo assay showed that leukocytes isolated from WT-HFD, but not from TKO-HFD, induced leukostasis and cell death. At 18 weeks, HFD triggered development of degenerated (acellular) capillaries and decreased branching density in WT but not in TKO mice. Together, HFD-induced obesity triggered early retinal leukostasis and microvascular dysfunction at least in part via TXNIP-NLRP3-inflammasome activation.


Effect of nitrogen intake on nitrogen recycling and urea transporter abundance in lambs.
Urea recycling in ruminants has been studied extensively in the past, but the mechanisms regulating the amount of urea recycled or excreted remain obscure. To elucidate the role of urea transporters (UT) in N recycling, nine Dorset-Finn ewe lambs (20.8 +/- 0.8 kg) were fed diets containing 15.5, 28.4, and 41.3 g of N/kg of DM for 25 d. Nitrogen balance and urea N kinetics were measured during the last 3 d of the period. Animals were then slaughtered and mucosa samples from the rumen, duodenum, ileum, and cecum, as well as kidney medulla and liver, were collected. Increasing N intake tended to increase N balance quadratically (1.5, 5.1, and 4.4 +/- 0.86 g of N/d, P < 0.09), and linearly increased urinary N excretion (2.4, 10, and 16.5 +/- 0.86 g N/d, P < 0.001) and plasma urea N concentration (4.3, 20.3, and 28.4 +/- 2.62 mg of urea N/dL, P < 0.001), but did not affect fecal N excretion (5.0 +/- 0.5 g of N/d; P < 0.94). Urea N production (2.4, 11.8, and 19.2 +/- 0.83 g of N/d; P < 0.001) and urinary urea N excretion (0.7, 7.0, and 13.4 +/- 0.73 g N/d; P < 0.001) increased linearly with N intake, as well as with the urea N recycled to the gastrointestinal tract (1.8, 4.8, and 5.8 +/- 0.40 g of N/d, P < 0.001). No changes due to N intake were observed for creatinine excretion (518 +/- 82.4 mg/d; P < 0.69) and clearance (46 +/- 10.7 mL/min; P < 0.56), but urea N clearance increased linearly with N intake (14.9, 24.4, and 34.9 +/- 5.9 mL/min; P < 0.04). Urea N reabsorption by the kidney tended to decrease (66.3, 38.5, 29.1 +/- 12.6%; P < 0.06) with increasing N content of the diet. Increasing the level of N intake increased linearly the weight of the liver as a proportion of BW (1.73, 1.88, and 2.22 +/- 0.15%, P < 0.03) but only tended to increase the weight of the kidneys (0.36, 0.37, and 0.50 +/- 0.05%, P < 0.08). Urea transporter B was present in all the tissues analyzed, but UT-A was detected only in kidney medulla, liver, and duodenum. Among animals on the three diets, no differences (P > 0.10) in UT abundance, quantified by densitometry, were found. Ruminal-wall urease activity decreased linearly (P < 0.02) with increasing level of N intake. Urease activity in duodenal, ileal, and cecal mucosa did not differ from zero (P > 0.10) in lambs on the high-protein diet. In the present experiment, urea transporter abundance in the kidney medulla and the gastrointestinal tract did not reflect the increase in urea-N reabsorption by the kidney and transferred into the gut.


Cultural determinants influence assisted reproduction usage in Europe more than economic and demographic factors.
To what extent do financial, demographic and cultural determinants explain the vast cross-national differences in ART treatments in Europe? The normative cultural acceptance of ART is a major driver of ART treatments in Europe, above and beyond differences in country wealth, demographic aspects and religious composition. There are vast differences in the number of ART treatments across European countries, which are to some extent related to country affluence, regulation, and insurance coverage and costs. The role and impact of cultural and normative factors has not been explored in a larger cross-national comparison. A descriptive and comparative cross-national analysis of ART treatment prevalence in over 30 European countries in 2010, with the outcome defined as the total number of ART cycles per million women of reproductive age (15-44 years). Data is drawn from multiple sources (ICMART, US Census Bureau Library, World Bank, Barro-Lee Educational Attainment Dataset, IFFS Surveillance reports, European Values Study and World Religion Database). Our sample includes data from 35 European countries, where we describe the associations between demographic and cultural factors and the prevalence of ART treatments. Bivariate correlation and ordinary least squares multiple regression analysis serves to establish the relationships between predictor variables and the number of ART treatments per million women aged 15-44 years in a country. A one-percent increase in national GDP is associated with 382 (95% CI: 177-587) additional ART procedures per million women of reproductive age, yet this effect is reduced to 99 (-92 to 290) treatments once cultural values and demographic factors are accounted for. In our fully adjusted model, normative cultural values measuring the acceptability of ART are the strongest predictor of ART usage, with a one-point increase of average approval in a country associated with 276 (167-385) additional ART treatments per million women of reproductive age. Findings are based on a cross-sectional, cross-national analysis, making formal tests of causality impossible and prohibiting inferences to the individual level. Results indicate that reproductive health policy should openly acknowledge the importance of cultural norms in informally shaping and regulating the wider availability of ART treatment. Funding for this project was provided by the European Union's Seventh Framework Program (FP7 2007-2013) (No. 320116 Families and Societies), European Research Council for the SOCIOGENOME Consolidator Grant (ERC-2013-CoG-615603) and the Wellcome Trust Institutional Strategic Support Fund (all to M.C.M.). The authors have no conflicts of interest to declare. N/A.


Usefulness of growth hormone (GH) stimulation tests and IGF-I concentration measurement in GH deficiency diagnosis.
The diagnosis of growth hormone (GH) deficiency (GHD) is still problematic for the clinician. There is no gold standard for estimating GH secretion. The aim of this study was to compare the diagnostic usefulness of spontaneous GH secretion test, pharmacological tests with insulin, clonidine, L-dopa, and glucagon, and IGF-I measurement in GHD. We studied 180 prepubertal, short children. Predictive values were calculated for different GH cutoff levels for each diagnostic test. ROC curves were used to estimate the diagnostic usefulness of the tests. The results show that sleep is the strongest stimulatory agent for GH secretion. The estimation of GH secretion after onset of sleep can be used as a screening test in GHD diagnosis. The insulin test has the highest discrimination. A combination of insulin test with another provocative test allows high discrimination and accuracy for standard cut-off GH level. Measurement of IGF-I is characterized by low predictive values. IGF-I level below the mean according to age indicates high probability of GHD. Auxological parameters should be the most important factor in diagnosing GHD.


Cyclic AMP-dependent positive feedback signaling pathways in the cortex contributes to visceral pain.
Cortical areas including the anterior cingulate cortex (ACC) play critical roles in different types of chronic pain. Most of previous studies focus on the sensory inputs from somatic areas, and less information about plastic changes in the cortex for visceral pain. In this study, chronic visceral pain animal model was established by injection with zymosan into the colon of adult male C57/BL6 mice. Whole cell patch-clamp recording, behavioral tests, western blot, and Cannulation and ACC microinjection were employed to explore the role of adenylyl cyclase 1 (AC1) in the ACC of C57/BL6 and AC1 knock out mice. Integrative approaches were used to investigate possible changes of neuronal AC1 in the ACC after the injury. We found that AC1, a key enzyme for pain-related cortical plasticity, was significantly increased in the ACC in an animal model of irritable bowel syndrome. Inhibiting AC1 activity by a selective AC1 inhibitor NB001 significantly reduced the up-regulation of AC1 protein in the ACC. Furthermore, we found that AC1 is required for NMDA GluN2B receptor up-regulation and increases of NMDA receptor-mediated currents. These results suggest that AC1 may form a positive regulation in the cortex during chronic visceral pain. Our findings demonstrate that the up-regulation of AC1 protein in the cortex may underlie the pathology of chronic visceral pain; and inhibiting AC1 activity may be beneficial for the treatment of visceral pain.


Assessment of Drug Resistance during Phase 2b Clinical Trials of Presatovir in Adults Naturally Infected with Respiratory Syncytial Virus.
This study summarizes drug resistance analyses in 4 recent phase 2b trials of the respiratory syncytial virus (RSV) fusion inhibitor presatovir in naturally infected adults. Adult hematopoietic cell transplant (HCT) recipients, lung transplant recipients, or hospitalized patients with naturally acquired, laboratory-confirmed RSV infection were enrolled in 4 randomized, double-blind, placebo-controlled studies with study-specific presatovir dosing. Full-length RSV F sequences amplified from nasal swabs obtained at baseline and postbaseline were analyzed by population sequencing. Substitutions at RSV fusion inhibitor resistance-associated positions are reported. Genotypic analyses were performed on 233 presatovir-treated and 149 placebo-treated subjects. RSV F variant V127A was present in 8 subjects at baseline. Population sequencing detected treatment-emergent substitutions in 10/89 (11.2%) HCT recipients with upper and 6/29 (20.7%) with lower respiratory tract infection, 1/35 (2.9%) lung transplant recipients, and 1/80 (1.3%) hospitalized patients treated with presatovir; placebo-treated subjects had no emergent resistance-associated substitutions. Subjects with substitutions at resistance-associated positions had smaller decreases in viral load during treatment relative to those without, but they had similar clinical outcomes. Subject population type and dosing regimen may have influenced RSV resistance development during presatovir treatment. Subjects with genotypic resistance development had decreased virologic responses compared to those without genotypic resistance but had comparable clinical outcomes.


Effect of brominated vegetable oils on heart lipid metabolism.
Normal rats fed for 105 days on an experimental diet made up of standard laboratory chow supplemented with 0.5% of a mixture of brominated sunflower-olive oil (BVO) developed a significant increase in the triacylglycerol content of the heart, liver and soleus muscle compared to controls. In addition, BVO-treated rats had a decrease in plasma levels of triacylglycerol and total and HDL cholesterol. Plasma fatty acid levels and plasma post-heparin lipolytic activities, such as H-TGL, LPL, T-TGL and MGH were similar to those of control animals fed the standard chow alone. Heart PDHa (active portion of pyruvate dehydrogenase) was dramatically decreased in the BVO-fed rats. A faster rate of spontaneous lipolysis was recorded in the isolated perfused preparation of hearts from the experimental animals. The addition of 10(-7) M of glucagon to the perfusate, however, revealed a lipolytic effect comparable to the one observed in the control rats. In summary, our findings of normal fatty acids and low triacylglycerol plasma levels associated with normal activities of the various PHLA (post-heparin lipolytic activity) enzymes suggest that accumulation of triacylglycerol in heart muscle may not be explained essentially in terms of an elevated uptake and/or increased delivery of plasma fatty acids or plasma triacylglycerol. A decreased in situ catabolism of tissue triacylglycerol also appears unlikely because the spontaneous as well as the glucagon induced lipolysis in the heart both were found to be unimpaired.(ABSTRACT TRUNCATED AT 250 WORDS)


Silver Nanoparticles Stimulates Spermatogenesis Impairments and Hematological Alterations in Testis and Epididymis of Male Rats.
The potential pharmaceutical application of nanoparticles has led to the toxicity within the male reproductive system. In the present study, the effects of silver nanoparticles (Ag-NPs) on hematological parameters, free radical generation, antioxidant system, sperm parameters, and organ histo-morphometry in male rats were investigated. Ag-NPs were produced by the reduction of silver ions, while the formation of which was monitored by UV-visible spectrophotometry. Zeta potential, transmission, and scanning electron microscopies were applied for the characterization of AgNPs. A total of 30 rats were divided into 6 groups and were sub-dermally exposed to Ag-NPs at the dosage of 0 (control), 10, and 50 mg/kg bodyweight (bw) doses for either 7 or 28 days. Ag-NP administration altered hematological indices and caused dose-dependent decreases in sperm motility, velocity, kinematic parameters, concentrations of luteinizing hormone, follicle-stimulating hormone, and testosterone. In the epididymis and testis, the concentrations of malondialdehyde and peroxide increases while superoxide dismutase, catalase, reduced glutathione, and total thiol group decreases. These findings suggest that Ag-NP triggered hormonal imbalance and induce oxidative stress in testis and epididymis; which negatively affect sperm parameters of male rats.


[The use of chronic epidural electrostimulation of the spinal cord in children with spastic diplegia--a type of infantile cerebral palsy].
The authors' first experience with spinal cord stimulation (SCS) at the lumbar level in the treatment of cerebral palsies (CP) in children aged 3-11 years who had lower spastic paraparesis of varying severity is presented. Complex clinical and physiological studies of all 9 children were made before and after surgery (follow-ups lasting 6-12 months). Preliminary results have indicated that the most significant changes occur primarily in pathological postural tonic reflexes. SCS has virtually no effect on fixed limb deformities, which requires further orthopedic surgical correction. A medical care schedule for CP children affected predominantly with lower spastic paraparesis should include, among its first priorities, electrode implantation for SCS. Due to the specific rehabilitative dynamics of motor functions, SCS should be made to treat these children before the deformities in the lower extremity joints get fixed.


BBA52 facilitates Borrelia burgdorferi transmission from feeding ticks to murine hosts.
Borrelia burgdorferi, the pathogen of Lyme borreliosis, persists in nature through a tick-rodent transmission cycle. A selective assessment of the microbial transcriptome, limited to gene-encoding putative membrane proteins, reveals that bba52 transcription in vivo is strictly confined to the vector-specific portion of the microbial life cycle, with the highest levels of expression noted in feeding ticks and with swift down-regulation noted in mice. bba52 deletion did not affect murine disease as assessed by the genesis of arthritis and carditis or long-term persistence of pathogens in mice or ticks. However, bba52 deficiency did impair microbial transitions between hosts and vector, defects that could be fully rescued when bba52 expression was genetically restored to the original genomic locus. These studies establish that BBA52 facilitates vector-host transitions by the pathogen and therefore is a potential antigenic target for interference with transmission of B. burgdorferi from ticks to mammalian hosts.


Early Prediction of Wound Healing Outcome Based on Chronic Wound Registry Database.
Chronic wounds cause a number of unnecessary amputations due to a delay in proper treatment. To expedite timely treatment, this paper presents an algorithm which uses a logistic regression classifier to predict whether the wound will heal or not within a specified time. The prediction is made at three time-points: one month, three months, and six months from the first visit of the patient to the healthcare facility. This prediction is made using a systematically collected chronic wound registry and is based entirely on data collected during patients' first visit. The algorithm achieves an area under the receiver operating characteristic curve (AUC) of 0.75, 0.72, and 0.71 for the prediction at the three time-points, respectively.Clinical relevance- Using the proposed prediction model, the clinicians will have an early estimate of the time taken to heal thereby providing appropriate treatments. We hope this will ensure timely treatments and reduce the number of unnecessary amputations.


Hydrostatic isolated limb perfusion with adeno-associated virus vectors enhances correction of skeletal muscle in Pompe disease.
Glycogen storage disease type II (Pompe disease; MIM 232300) stems from the inherited deficiency of acid-α-glucosidase (GAA; acid maltase; EC 220.127.116.11), which primarily involves cardiac and skeletal muscles. We hypothesized that hydrostatic isolated limb perfusion (ILP) administration of an adeno-associated virus (AAV) vector containing a muscle-specific promoter could achieve relatively higher transgene expression in the hindlimb muscles of GAA-knockout (GAA-KO) mice, in comparison with intravenous (IV) administration. ILP administration of AAV2/8 vectors encoding alkaline phosphatase or human GAA-transduced skeletal muscles of the hindlimb widely, despite the relatively low number of vector particles administered (1 × 10¹¹), and IV administration of an equivalent vector dose failed to transduce skeletal muscle detectably. Similarly, ILP administration of fewer vector particles of the AAV2/9 vector encoding human GAA (3 × 10¹⁰) transduced skeletal muscles of the hindlimb widely and significantly reduced glycogen content to, in comparison with IV administration. The only advantage for IV administration was moderately high-level transduction of cardiac muscle, which demonstrated compellingly that ILP administration sequestered vector particles within the perfused limb. Reduction of glycogen storage in the extensor digitorum longus demonstrated the potential advantage of ILP-mediated delivery of AAV vectors in Pompe disease, because type II myofibers are resistant to enzyme replacement therapy. Thus, ILP will enhance AAV transduction of multiple skeletal muscles while reducing the required dosages in terms of vector particle numbers.


Rapid counting of liquid-borne microorganisms by light scattering spectrometry.
Fast and sensitive techniques are needed to determine microorganism presence in liquid samples. In this research, the feasibility of using light scattering spectrometry for enumerating the biological particles in liquid samples was investigated. A particle size spectrometer was used to count six commonly found microbial species suspended in liquid with and without microbiological stains applied: Pseudomonas fluorescens, Micrococcus spp. vegetative cells and Bacillus subtilis var. niger endospores were stained with Acridine Orange and Crystal Violet, while Cladosporium cladosporioides, Penicillium melinii and Aspergillus versicolor fungi were stained with Acridine Orange and Lactophenol Cotton Blue. The counts obtained with the spectrometer were compared with those obtained with a phase-contrast microscope. It was found that the spectrometer counted about 32 % of non-stained B. subtilis endospores and this percentage increased to almost 90 % for stained endospores. Among the investigated species of fungi, the counting efficiency of P. melinii was the only one significantly affected by the application of the stain Lactophenol Cotton Blue: the fraction of counted fungal spores increased from 64 % (non-stained spores) to about 100 % (stained spores). The observed difference in counting efficiency may serve as a basis for differentiating biological from non-biological particles in liquid samples.


[Epidemiology of severe chronic renal insufficiency in Burkina Faso].
We identified 174 cases of chronic severe renal failure (blood creatinine > 650 mumol/l) and/or blood urea > 35 mmol/l) in a retrospective study of patients admitted to hospital between January 1989 and June 1996. Of these patients, 110 were men and 64 were women. The mean age was 36 +/- 15 years. Fifty three patients had a history of hypertension before admission, 3 patients had diabetes and 3 had gout. The most frequent clinical signs were dyspnea (55.2% of all patients), fatigue (78.2%), vomiting (63.2%) and edema (66.1%). The prevalence of hypertension was 64.9%. Glomerulonephritis was found in 42.5% of patients, chronic interstitial nephritis in 16.1%, polycystic kidney disease in 2 cases, congenital renal hypoplasia in 4 cases and unclassified kidney disease in 14.4% of cases. End-stage renal failure was complicated by heart failure in 40.2% of patients, pericarditis in 31.6%, hemorrhage of the gastrointestinal tract in 15% and infections in 22.4%. 47.7% of the patients died following admission.


Alpha 1-adrenergic stimulation of rat myocardial cells increases protein synthesis.
The effects of adrenergic stimulation on the rates of protein synthesis, degradation, and accumulation were examined in primary cultures of neonatal rat heart cells. Treatment of myocardial cells with norepinephrine increased total cellular protein content and the rate of incorporation of radiolabeled tyrosine into trichloroacetic acid insoluble protein. alpha 1-Adrenergic, but not alpha 2- or beta-adrenergic blockade, inhibited these norepinephrine induced increases. The rate of protein synthesis estimated from the kinetics of equilibrium labeling and from combined equilibrium and pulse labeling was increased by norepinephrine stimulation, whereas protein degradation estimated by release of previously incorporated radiolabeled tyrosine or in pulse-chase experiments was unaffected. To determine whether alpha 1-adrenergic stimulation produced similar effects on the turnover of myofibrillar proteins, rates of synthesis and degradation were estimated for a myofibrillar-enriched protein fraction and for myosin heavy chain and actin. Norepinephrine treatment produced increases in the synthesis of myofibrillar protein without significantly altering degradation rates. These experiments suggest that alpha 1-adrenergic stimulation increases myocardial cell protein content by accelerating protein synthesis.


Surface enhanced coherent anti-stokes Raman scattering on nanostructured gold surfaces.
Coherent anti-Stokes Raman spectroscopy (CARS) is a well-known tool in multiphoton imaging and nonlinear spectroscopy. In this work we combine CARS with plasmonic surface enhancement on reproducible nanostructured surfaces. We demonstrate strong correlation between plasmon resonances and surface-enhanced CARS (SECARS) intensities on our nanostructured surfaces and show that an enhancement of ∼10(5) can be obtained over standard CARS. Furthermore, we find SECARS to be >10(3) times more sensitive than surface-enhanced Raman Spectroscopy (SERS). We also demonstrate SECARS imaging of molecular monolayers. Our work paves the way for reliable single molecule Raman spectroscopy and fast molecular imaging on plasmonic surfaces.


Pharmacokinetic interaction of megestrol acetate with zidovudine in human immunodeficiency virus-infected patients.
This nonrandomized, two-period crossover study was performed to assess whether concomitant administration of megestrol acetate influences the steady-state pharmacokinetics of zidovudine and its inactive 5'-O-glucuronide metabolite. Twelve HIV-positive, asymptomatic male volunteers received a 100-mg oral capsule dose of zidovudine at least 30 min before meals five times a day at 0700, 1100, 1500, 1900, and 2300 h on study days 1 to 3 and a single 100-mg dose at 0700 h on day 4. On days 5 to 17, 800 mg of megestrol acetate, as a 40-mg/ml aqueous suspension, was administered orally immediately before the 0700 h dose of zidovudine. On days 5 to 16, zidovudine was also administered at 1100, 1500, 1900, and 2300 h. Serial blood samples were collected for 12 h after the single 100-mg dose of zidovudine on days 4 and 17; trough samples were also obtained just before the 0700 h dose on days 2 to 4 and 15 to 17. Levels of zidovudine and its glucuronide in plasma were assayed by a validated radioimmunoassay. Statistical analysis of trough plasma level data indicated that steady-state levels of zidovudine and its glucuronide in plasma had been attained when pharmacokinetic assessments were made on days 4 and 17. When megestrol acetate and zidovudine were coadministered for 13 days, differences of -14, -6.5, and -4.6% in mean zidovudine peak concentration and areas under the curve at 0 to 4 and 0 to 12 h, respectively, +22.5% in mean trough concentration, +2.6% in mean plasma half-life, and no change in median time to peak were observed compared to conditions when zidovudine was administered alone; for zidovudine 5'-O-glucuronide the respective differences were -9, -7.3, -4.4, +2.3, and +10% and no change. None of the differences were statistically significant (P > 0.05). Concomitant therapy with megestrol acetate, at the dose employed to treat anorexia, cachexia, or an unexplained, significant weight loss in AIDS patients, did not alter the steady-state pharmacokinetics of zidovudine or its 5'-O-glucuronide metabolite.


Structural basis of matrix metalloproteinase function.
The matrix metalloproteinases (MMPs) constitute a family of multidomain zinc endopeptidases which contain a catalytic domain with a common metzincin-like topology. The MMPs are involved not only in extracellular matrix degradation, but also in a number of other biological processes. Normally, their proteolytic activity is regulated precisely by their main endogenous protein inhibitors, in particular the tissue inhibitors of metalloproteinases (TIMPs). Disruption of this balance results in serious diseases, such as arthritis, tumour growth and metastasis, rendering the MMPs attractive targets for inhibition therapy. Knowledge of their tertiary structures is crucial for a full understanding of their functional properties. Since the first publication of atomic MMP structures in 1994, much more structural information has become available on details of the catalytic domain, on its interaction with synthetic and protein inhibitors, on domain organization and on the formation of complexes with other proteins. This review will outline our current knowledge of MMP structure.


Memory bias for schema-related stimuli in individuals with bulimia nervosa.
This study investigates whether individuals with bulimia nervosa (BN) have a memory bias in relation to explicit memory (cued and free recall vs. verbal and pictorial recognition tasks). Twenty-five participants diagnosed with BN and 27 normal controls (NC) were exposed to body-related, food-related, and neutral TV commercials, and then recall and recognition rates were assessed. Poorer recognition and recall of body-related stimuli was found for BN in comparison to NC, suggesting a memory bias. Results are discussed in relation to previous studies, along with suggestions as to how future studies can gain more insight into dysfunctions in information processing that can lead to the maintenance of eating disorders.


Pyogranulomatous mural folliculitis in a cat treated with methimazole.
An 11-year-old spayed female domestic shorthair cat was presented for polydipsia, hyperactivity and bilateral thyroid gland enlargement. Total T4 (TT4) was in the upper interval range; therefore, an early hyperthyroidism was suspected. A treatment trial with methimazole was started, as the owner refused further tests. Six months later the owner stopped the treatment. One year later, clinical signs persisted and TT4 was still in the upper interval range. Methimazole was re-introduced but 48 h later the cat presented non-pruritic alopecia with erythema, scales and perilesional yellowish crusts. Pyogranulomatous mural folliculitis was diagnosed by histopatological examination of the skin biopsies. Methimazole was withdrawn and macroscopic lesions healed and disappeared histologically in 15 days. An idiosyncratic drug reaction to methimazole was suspected. To the best of our knowledge, this is the first report of feline pyogranulomatous mural folliculitis likely secondary to an adverse drug reaction to methimazole administration.


First Report of Coexistence of Two Ectopic Pituitary Tumors: Rathke Cleft Cyst and Silent Adrenocorticotropic Hormone Adenoma.
Rathke cleft cysts (RCCs) and pituitary adenomas (PAs) are thought to have a common embryonic ancestry; however, PAs with a concomitant RCC inside the sella turcica are rarely observed. Ectopic pituitary tumors are also rare. We present the case of a 65-year-old woman with an ectopic RCC in the sphenoid sinus and outside the sella turcica concomitant with an adrenocorticotropic hormone (ACTH)-staining, clinically silent PA. The patient had headache but no endocrine or visual disturbances. Preoperative magnetic resonance imaging revealed infrasellar cystic lesion in the sphenoid sinus with erosion of the clivus and intact sellar floor. The patient underwent gross total microsurgical resection through the transnasal route with an uneventful postoperative course. To our knowledge, this is the first reported ectopic RCC located outside the sella turcica with a concomitant ACTH-staining PA. This also appears to be the first ACTH-staining adenoma concomitant with RCC reported in the literature, regardless of location, not presenting with Cushing disease. This case shows that we can now include pituitary adenoma with or without a concomitant RCC in the differential diagnosis of processes in the sphenoid sinus. As both PAs and RCCs are benign sellar lesions, surgical management of a concomitant occurrence of these tumors mainly depends on the size of the lesions and their clinical manifestations. For patients with PA and concomitant RCC, surgical resection should be considered, as there is an approximatrely 20% recurrence rate of the cyst after resection and the possibility of future clival erosion, if left untreated.


Assessing content validity and user perspectives on the Team Check-up Tool: expert survey and user focus groups.
This study assesses content validity and user feedback on the Team Check-up Tool (TCT), an instrument used for measuring dynamic context of quality improvement (QI) teams and their implementation of QI activities. We conducted two focus groups and one larger feedback session with TCT users to assess feasibility, importance of areas of inquiry and barriers to use. A panel of eight QI experts evaluated the item-by-item content (content validity) of TCT by rating the relevance of each item to implementation success. We calculated item-level and scale-level content validity using the content validity index (CVI). Scale-level CVI was 0.872. Highly rated items included implementation of recommended interventions, educational activities, team review of performance data, team sharing of performance data with staff and specific barriers to progress. Four items were rated relatively low: presentation of performance data to the hospital/health system board; manner of provision of feedback of data to staff; to what other units the team attempted to spread and turnover of QI team members. Items identified in user focus groups as important included whether there were events distracting staff from the initiative, number of team meetings and turnover of QI team members. Focus groups also identified barriers to the completion of the tool, including lack of feedback and response fatigue during stable activity periods. The findings support the conclusion that the TCT measures meaningful areas of context and implementation in team-based QI initiatives, particularly intervention activity tracking, review and sharing of performance data and team progress barriers. We offer a modified instrument with a framework for real-time measurement of important elements of implementation and context of QI teams based on the findings.


Crystal structures of di-bromido-{N-[(pyridin-2-yl-κN)methyl-idene]picolinohydrazide-κ2N',O}cadmium methanol monosolvate and di-iodido{N-[(pyridin-2-yl-κN)methyl-idene]picolinohydrazide-κ2N',O}cadmium.
The title compounds, [CdBr2(C12H10N4O)]·CH3OH, (I), and [CdI2(C12H10N4O)], (II), are cadmium bromide and cadmium iodide complexes of the ligand (E)-N'-(pyridin-2-yl-methyl-ene)picolinohydrazide. Complex (I) crystallizes as the methanol monosolvate. In both compounds, the Cd2+ cation is ligated by one O atom and two N atoms of the tridentate ligand, and by two bromide anions forming a Br2N2O penta-coordination sphere for (I), and by two iodide anions forming an I2N2O penta-coordination sphere for (II), both with a distorted square-pyramidal geometry. In the crystal of complex (I), mol-ecules are linked by pairs of N-H⋯O and O-H⋯Br hydrogen bonds, involving the solvent mol-ecule, forming dimeric units, which are linked by C-H⋯Br hydrogen bonds forming layers parallel to (101). In the crystal of complex (II), mol-ecules are linked by N-H⋯I hydrogen bonds, forming chains propagating along [010]. In complex (II), measured at room temperature, the two iodide anions are each disordered over two sites; the refined occupancy ratio is 0.75 (2):0.25 (2).


Acquired prosopagnosia with structurally intact and functional fusiform face area and with face identity-specific configuration processing deficits.
Prosopagnosia or loss of face perception and recognition is still poorly understood and rare single cases of acquired prosopagnosia can provide a unique window on the behavioural and brain basis of normal face perception. The present study of a new case of acquired prosopagnosia with bilateral occipito-temporal lesions but a structurally intact FFA and OFA investigated whether the lesion overlapped with the face network and whether the structurally intact FFA showed a face selective response. We also investigated the behavioral correlates of the neural findings and assessed configural processing in the context of facial and non-facial identity recognition, expression recognition and memory, also focusing on the face-selectivity of each specific deficit. The findings reveal a face-selective response in the FFA, despite lesions in the face perception network. At the behavioural level, the results showed impaired configural processing for facial identity, but not for other stimulus categories and not for facial expression recognition. These findings challenge a critical role of the FFA for face identity processing and support a domain-specific account of configural processing.


Characterization of type-2 diacylglycerol acyltransferases in Haematococcus lacustris reveals their functions and engineering potential in triacylglycerol biosynthesis.
Haematococcus lacustris is an ideal source of astaxanthin (AST), which is stored in oil bodies containing esterified AST (EAST) and triacylglycerol (TAG). Diacylglycerol acyltransferases (DGATs) catalyze the last step of acyl-CoA-dependent TAG biosynthesis and are also considered as crucial enzymes involved in EAST biosynthesis in H. lacustris. Previous studies have identified four putative DGAT2-encoding genes in H. lacustris, and only HpDGAT2D allowed the recovery of TAG biosynthesis, but the engineering potential of HpDGAT2s in TAG biosynthesis remains ambiguous. Five putative DGAT2 genes (HpDGAT2A, HpDGAT2B, HpDGAT2C, HpDGAT2D, and HpDGAT2E) were identified in H. lacustris. Transcription analysis showed that the expression levels of the HpDGAT2A, HpDGAT2D, and HpDGAT2E genes markedly increased under high light and nitrogen deficient conditions with distinct patterns, which led to significant TAG and EAST accumulation. Functional complementation demonstrated that HpDGAT2A, HpDGAT2B, HpDGAT2D, and HpDGAT2E had the capacity to restore TAG synthesis in a TAG-deficient yeast strain (H1246) showing a large difference in enzymatic activity. Fatty acid (FA) profile assays revealed that HpDGAT2A, HpDGAT2D, and HpDGAT2E, but not HpDGAT2B, preferred monounsaturated fatty acyl-CoAs (MUFAs) for TAG synthesis in yeast cells, and showed a preference for polyunsaturated fatty acyl-CoAs (PUFAs) based on their feeding strategy. The heterologous expression of HpDGAT2D in Arabidopsis thaliana and Chlamydomonas reinhardtii significantly increased the TAG content and obviously promoted the MUFAs and PUFAs contents. Our study represents systematic work on the characterization of HpDGAT2s by integrating expression patterns, AST/TAG accumulation, functional complementation, and heterologous expression in yeast, plants, and algae. These results (1) update the gene models of HpDGAT2s, (2) prove the TAG biosynthesis capacity of HpDGAT2s, (3) show the strong preference for MUFAs and PUFAs, and (4) offer target genes to modulate TAG biosynthesis by using genetic engineering methods.


Comparison of Risk of Silicosis in Metal Mines and Pottery Factories: A 44-Year Cohort Study.
Little is known about the different risk of silicosis in metal mines and pottery factories. We aimed to compare the silicosis risks among silica-exposed workers in different industrial circumstances. Are the silicosis risks among silica-exposed workers in industrial circumstances different? We studied 39,808 workers followed up from January 1, 1960 to December 31, 2003 in China. Cumulative respirable silica dust exposure (CDE) was estimated by linking a job-exposure matrix to personal work history. Silicosis of stage I or higher was diagnosed by Chinese pneumoconiosis Roentgen diagnostic criteria. A total of 9,377 silicosis patients were diagnosed during 1,153,580.9 person-years' follow-up in the cohort. Hazard ratios of silicosis for each 1 mg/m3-year increase in CDE were 1.08 (1.07-1.08) for tungsten mines, 1.41 (1.33-1.48) for iron and copper mines, 1.14 (1.11-1.17) for tin mines, and 1.03 (1.02-1.04) for pottery factories, respectively. When exposed to 0.05 mg/m3 of respirable silica dust for 45 years, the cumulative risks in metal mines (2.3%, 9.9%, 1.5% for tungsten mines, iron and copper mines, and tin mines, respectively) were still higher than those in pottery factories (0.6%). The joint effect of silica and smoking on silicosis was more than multiplicative. The risk of silicosis in metal miners is higher than that in pottery workers when exposed to the same level of silica dust. The silica dust exposed years should be under 10 years for metal miners and 40 years for pottery workers at 0.05 mg/m3 to keep lifetime risk within 0.1%. Current exposure limits should take into account differences in various industrial circumstances. Smoking cessation could help reduce silicosis risk for silica-exposed workers.


Determination of alpha-2-macroglobulin complexes by a new immuno-activity assay.
Alpha-2-macroglobulin (α(2)M) is a broad specificity protease inhibitor which impacts several hemostatic pathways. Selective detection of various α(2)M complexes may be useful to define markers for the status of different hemostatic components. We present proof of principle for a novel assay to quantitatively measure α(2)M in complex with a variety of hemostatic factors. The assay makes use of the fact that α(2)M entraps proteases within a molecular "cage", leaving them inaccessible to macromolecular substrates while retaining functionality against small synthetic substrates. Wells coated with anti-α(2)M antibodies were used to isolate the complexes from buffer or plasma, followed by detection of specific proteases with chromogenic substrates. Macromolecular inhibitors were added to eliminate signal from any unbound proteases. Calibration curves constructed with purified protease-α(2)M complexes were sigmoidal in nature, as is typical with immuno-assays. The specificity of signal production was confirmed with inhibitors that target either free protease, or both free and α(2)M-bound protease. The detection range of the assay was dependent on the protease being measured, and the surrounding matrix. Interference in detection of complexes in plasma was found to be caused, in part, by free α(2)M. Thrombin-α(2)M complexes were quantified in adult and newborn plasma following induction of thrombin generation and found to be significantly higher in adults, likely due to higher prothrombin levels. This assay provides a versatile platform method for quantification of multiple protease-α(2)M complexes. It may prove useful for mechanistic in vitro studies of hemostatic pathways, and potentially for clinical applications.


High-resolution, small animal radiation research platform with x-ray tomographic guidance capabilities.
To demonstrate the computed tomography, conformal irradiation, and treatment planning capabilities of a small animal radiation research platform (SARRP). The SARRP uses a dual-focal spot, constant voltage X-ray source mounted on a gantry with a source-to-isocenter distance of 35 cm. Gantry rotation is limited to 120 degrees from vertical. X-rays of 80-100 kVp from the smaller 0.4-mm focal spot are used for imaging. Both 0.4-mm and 3.0-mm focal spots operate at 225 kVp for irradiation. Robotic translate/rotate stages are used to position the animal. Cone-beam computed tomography is achieved by rotating the horizontal animal between the stationary X-ray source and a flat-panel detector. The radiation beams range from 0.5 mm in diameter to 60 x 60 mm(2). Dosimetry is measured with radiochromic films. Monte Carlo dose calculations are used for treatment planning. The combination of gantry and robotic stage motions facilitate conformal irradiation. The SARRP spans 3 ft x 4 ft x 6 ft (width x length x height). Depending on the filtration, the isocenter dose outputs at a 1-cm depth in water were 22-375 cGy/min from the smallest to the largest radiation fields. The 20-80% dose falloff spanned 0.16 mm. Cone-beam computed tomography with 0.6 x 0.6 x 0.6 mm(3) voxel resolution was acquired with a dose of <1 cGy. Treatment planning was performed at submillimeter resolution. The capability of the SARRP to deliver highly focal beams to multiple animal model systems provides new research opportunities that more realistically bridge laboratory research and clinical translation.


The Cox protein is a modulator of directionality in bacteriophage P2 site-specific recombination.
The P2 Cox protein is known to repress the Pc promoter, which controls the expression of the P2 immunity repressor C. It has also been shown that Cox can activate the late promoter PLL of the unrelated phage P4. By this process, a P2 phage infecting a P4 lysogen is capable of inducing replication of the P4 genome, an example of viral transactivation. In this report, we present evidence that Cox is also directly involved in both prophage excision and phage integration. While purified Cox, in addition to P2 Int and Escherichia coli integration host factor, was required for attR x attL (excisive) recombination in vitro, it was inhibitory to attP x attB (integrative) recombination. The same amounts of Int and integration host factor which mediated optimal excisive recombination in vitro also mediated optimal integrative recombination. We quantified and compared the relative efficiencies of attB, attR, and attL in recombination with attP and discuss the functional implications of the results. DNase I protection experiments revealed an extended 70-bp Cox-protected region on the right arm of attP, centered at about +60 bp from the center of the core sequence. Gel shift assays suggest that there are two Cox binding sites within this region. Together, these data support the theory that in vivo, P2 can exert control over the direction of recombination by either expressing Int alone or Int and Cox together.


A Hybrid alldifferent-Tabu Search Algorithm for Solving Sudoku Puzzles.
The Sudoku problem is a well-known logic-based puzzle of combinatorial number-placement. It consists in filling a n(2) × n(2) grid, composed of n columns, n rows, and n subgrids, each one containing distinct integers from 1 to n(2). Such a puzzle belongs to the NP-complete collection of problems, to which there exist diverse exact and approximate methods able to solve it. In this paper, we propose a new hybrid algorithm that smartly combines a classic tabu search procedure with the alldifferent global constraint from the constraint programming world. The alldifferent constraint is known to be efficient for domain filtering in the presence of constraints that must be pairwise different, which are exactly the kind of constraints that Sudokus own. This ability clearly alleviates the work of the tabu search, resulting in a faster and more robust approach for solving Sudokus. We illustrate interesting experimental results where our proposed algorithm outperforms the best results previously reported by hybrids and approximate methods.


Acute oral tetrahydrobiopterin administration ameliorates endothelial dysfunction in systemic sclerosis.
Systemic sclerosis (SSc) is a rare, autoimmune disease characterised by endothelial dysfunction, which is associated with peripheral vasculopathy, such as digital ulcers (DU). We sought to determine if acute oral administration of tetrahydrobiopterin (BH4), an essential cofactor for endothelial nitric oxide synthase, would augment endothelial function in patients with SSc. Twelve SSc patients, of whom a majority had a history of DU, were studied 5 hours after oral BH4 administration (10 mg/kg body mass) or placebo on separate days using controlled, counterbalanced, double-blind, crossover experimental design. There were no differences in blood markers of oxidative stress and brachial artery blood pressure, diameter, blood velocity, shear rate, or blood flow at rest between placebo and BH4 (p>0.05). Whereas, after a 5 minute suprasystolic forearm cuff occlusion, brachial artery peak reactive hyperemia (placebo: 313±30 vs. BH4: 347±37 ml/min, p<0.05) and flow-mediated dilation (FMD) (placebo: 3.0±0.8 vs. BH4: 4.8±0.8%, p<0.05) were significantly higher after acute BH4 administration, indicating an improvement in endothelial function. To determine if the vasodilatory effects of BH4 were specific to the vascular endothelium, brachial artery blood flow and vasodilation in response to sublingual nitroglycerin were assessed, and were found to be unaffected by BH4 (p>0.05). These findings indicate that acute BH4 administration ameliorates endothelial dysfunction in patients with SSc. Given that endothelial dysfunction is known to be associated with DU in SSc patients, this study provides a proof-of-concept for the potential therapeutic benefits of BH4 in the prevention or treatment of DU in this population.