The D-dimer assay: a possible tool in the evaluation of atrial thrombosis.
Atrial fibrillation (AF) is a common arrhythmia seen in clinical practice, and affects more than 4% of the population older than 60 years of age. Peripheral thromboembolism contributes significantly to the observed morbidity and mortality. Symptomatic AF, before cardioversion to normal sinus rhythm, requires either exclusion of atrial thrombi using transesophageal echocardiography (TEE) or the conventional use of three weeks of adequate anticoagulation. The exclusion of atrial thrombi by TEE, a nontomographic technique but comparable with conventional treatment of AF in outcomes, has inherent limitations due to the complex three-dimensional multilobed anatomy of the left atrial appendage, where the majority of atrial thrombi arise. Also, the conventional treatment of three weeks of therapeutic anticoagulation before cardioversion reportedly does not always eliminate atrial thrombi. Plasma D-dimer constitutes an antigen-antibody reaction to the dimeric final degradation product of a mature clot. An elevated fibrin D-dimer has a high sensitivity for intravascular thrombosis and, hence, may improve the evaluation of a patient with AF before cardioversion in addition to a TEE. A case is presented in which a positive D-dimer resulted in performing TEE to document atrial thrombosis and the complications of previous bacterial endocarditis. In the present case, this involved aortic root abscess formation and acute aortic regurgitation because of flailing of the noncoronary cusp that resulted in recurrent pulmonary edema.


Local inhibition modulates odor-evoked synchronization of glomerulus-specific output neurons.
At the first stage of olfactory processing in the brain, synchronous firing across glomeruli may help to temporally bind multiple and spatially distributed input streams activated by a given odor. This hypothesis, however, has never been tested in an organism in which the odor-tuning properties of several spatially identifiable glomeruli are known. Using the sphinx moth, an insect that meets these specific criteria, we recorded odor-evoked responses simultaneously from pairs of projection neurons (PNs) innervating the same or different glomeruli in the macroglomerular complex (MGC), which is involved in processing pheromonal information. PNs that branched in the same glomerulus and were activated by the same pheromone component also showed the strongest coincident responses to each odor pulse. Glomerulus-specific PN pairs were also inhibited by the pheromone component that selectively activated PNs in the neighboring glomerulus, and about 70% of all intraglomerular pairs showed increased synchronization when stimulated with a mixture of the two odorants. Thus, when two adjacent glomeruli receive their inputs simultaneously, the temporal tuning of output from each glomerulus is enhanced by reciprocal and inhibitory interglomerular interactions.


Molecular biological evaluation of bioactive glass microspheres and adjunct bone morphogenetic protein 2 gene transfer in the enhancement of new bone formation.
Bioactive glass is a promising osteoconductive silica-based biomaterial for guidance of new bone growth. On the basis of several in vitro studies, the material appears able to promote osteoblast functions. In our in vivo study, the osteopromotive effect of bioactive glass microspheres seemed to surpass the osteoinductive action of direct adenovirus-mediated human bone morphogenetic protein 2 (BMP-2) gene transfer in a noncritical size bone defect model. The current study was initiated to elucidate the molecular mechanism behind bioactive glass action with or without adjunct BMP-2 gene transfer. A standardized bone defect of the rat tibia was filled with bioactive glass microspheres and injected with adenovirus carrying the human BMP-2 gene (RAdBMP-2). Control defects were left empty or filled with bioactive glass microspheres with injection of adenovirus carrying the lacZ reporter gene or saline. Quantitative polymerase chain reaction confirmed the expression of the transferred human BMP-2 gene at the defect area at 4 days, but not in intact reference tissues. Bone matrix components (collagens I, II, and III, osteocalcin, osteonectin, and osteopontin) and resorption markers (cathepsin K and MMP-9), determined by Northern analysis, showed a completely different pattern of gene expression in defects filled with bioactive glass compared with control defects left to heal without filling. Bioactive glass induced a long-lasting production of bone matrix with concurrent upregulation of osteoclastic markers, a sign of high bone turnover. Combining RAdBMP-2 gene transfer with bioactive glass decelerated the high turnover, but did not influence the balance of synthesis and resorption. This molecular analysis confirmed not only the highly osteopromotive effect of bioactive glass microspheres, but also the accelerated rate of new bone resorption on its surface. At least in noncritical size defects this impact of bioactive glass seems to saturate new bone formation on its surface and thereby overshadow the effect of BMP-2 gene transfer.


Displaced femoral neck fractures in the elderly: outcomes and cost effectiveness.
The optimal treatment for displaced femoral neck fractures in elderly patients is a matter of controversy. Four surgical options are well supported in the orthopaedic literature: reduction with internal fixation, unipolar hemiarthroplasty, bipolar hemiarthroplasty, and total hip arthroplasty. Based on a review of the outcomes literature regarding treatment of femoral neck fractures and a cost-effectiveness analysis, an algorithm for surgical treatment of displaced femoral neck fractures in elderly patients is presented. Cost-effectiveness analysis of these four surgical treatment options shows that arthroplasty is the most cost-effective treatment when complication rate, mortality, reoperation rate, and function are evaluated during a 2-year postoperative period. These data were strongly supported by a two-way sensitivity analysis that varied the effectiveness of the interventions and the costs. Literature derived outcome studies show that elderly patients with displaced femoral neck fractures achieve the best functional results with a well healed femoral neck without osteonecrosis after reduction and internal fixation. Achieving this result may be difficult, and it is not as cost effective as arthroplasty.


Effects of dietary fiber and feeding frequency on ruminal fermentation, digesta water-holding capacity, and fractional turnover of contents.
To determine the effects of different sources of fiber and feeding frequency on digesta water-holding capacity (WHC; g H2O/g DM) and ruminal liquid contents, four ruminally fistulated Jersey steers were fed a 60:40 roughage-concentrate diet at 1.5 times NEm. Diets contained either sorghum silage (SS) or a 67:33 mixture of SS and soyhulls (SH) as roughage and were fed either once or 12 times daily, in a 2 x 2 factorial experiment with 15-d periods. Ruminal fluid was sampled at 2, 4, 6, 8, 10, 12, 16, 20, and 24 h after a dose of Co-EDTA on d 10 and analyzed for Co, VFA, ammonia, buffering capacity, and osmolality. Ruminal WHC, NDF, ADF, lignin, and starch were measured in samples obtained by ruminal evacuation at 3, 6, 12, and 24 h after feeding on d 11, 12, 14, and 15, respectively. Substitution of SH for SS decreased ruminal pH .32 units and dilution rate by 26.8% but increased total VFA by 10.9%, osmolality by 13.6%, and the fractional turnover rate (FTR) of ADF by 22.5% (P less than .05). Frequent feeding resulted in 4.7, 21.9, and 74.4% increases in total VFA and FTR of ruminal DM and starch (P less than .05), respectively. Interactions (P less than .05) were observed between dietary fiber source and feeding frequency for ruminal fluid molar percentage acetate to propionate ratio (A/P), liquid volume (evacuated), and WHC (kilograms). Substituting SH for SS decreased ruminal WHC (kilograms), liquid volume, and A/P only in steers fed once daily. Ruminal WHC (kilograms) was correlated positively with ruminal liquid volume but negatively with DM FTR. The dynamics of digesta WHC (kilograms) associated with dietary fiber source and feeding frequency suggest that it may influence the contribution of water and salivary secretions to ruminal liquid contents.


Single-cell dissection of cellular components and interactions shaping the tumor immune phenotypes in ovarian cancer.
Distinct T cell infiltration patterns, i.e., immune infiltrated, excluded, and desert, result in different responses to cancer immunotherapies. However, the key determinants and biology underpinning these tumor immune phenotypes remain elusive. Here, we provide a high-resolution dissection of the entire tumor ecosystem through single-cell RNA-sequencing analysis of 15 ovarian tumors. Immune-desert tumors are characterized by unique tumor cell-intrinsic features, including metabolic pathways and low antigen presentation, and an enrichment of monocytes and immature macrophages. Immune-infiltrated and -excluded tumors differ markedly in their T cell composition and fibroblast subsets. Furthermore, our study reveals chemokine receptor-ligand interactions within and across compartments as potential mechanisms mediating immune cell infiltration, exemplified by the tumor cell-T cell cross talk via CXCL16-CXCR6 and stromal-immune cell cross talk via CXCL12/14-CXCR4. Our data highlight potential molecular mechanisms that shape the tumor immune phenotypes and may inform therapeutic strategies to improve clinical benefit from cancer immunotherapies.


Edge states of periodically kicked quantum rotors.
We present a quantum localization phenomenon that exists in periodically kicked three-dimensional rotors, but is absent in the commonly studied two-dimensional ones: edge localization. We show that under the condition of a fractional quantum resonance there are states of the kicked rotor that are strongly localized near the edge of the angular momentum space at J=0. These states are analogs of surface states in crystalline solids, and they significantly affect resonant excitation of molecular rotation by laser pulse trains.


Epsilon carbide: a low-temperature component of interplanetary dust particles.
Transmission electron microscope study of a chondritic interplanetary dust particle has revealed the presence of epsilon iron-nickel carbide, a low- temperature carbide previously encountered only in metallurgical studies. In these studies in-carbide was synthesized by carburization of iron or nickel grains in a stream of carbon monoxide or carbon monoxide plus hydrogen. Similar carburization of an iron-nickel metal in situ may have produced in-carbide during particle heating on atmospheric entry or in solar orbit. Alternatively, the epsilon-carbide may be a by-product of Fischer-Tropsch reactions in the solar nebula. Such reactions have been proposed as the mechanism of hydrocarbon formation in the early solar system.


Hypoestrogenism in young women and its influence on bone mass density.
One of the most important hormonal factors responsible for bone health is estradiol. Genetic factors, adequacy of hormonal functioning, nutrition and physical activity may be the markers of bone status and development in young women. During adolescence, women reach peak bone acquisition and develop a skeletal mass. This process is largely regulated by endocrine factors mainly such as adequate levels of gonadal, adrenal and pituitary hormones. The crucial role played by estradiol and its impact on bones are very multiple. Estradiol induces growth factors' activation, receptor activator of nuclear factor kappa B ligand (RANKL) production inhibition and is mainly referred to antiresorptive activity. Clinical situations leading to hypoestrogenism has been linked to decreased bone mineral density leading to osteopenia and osteoporosis. This status both in fertile and perimenopausal women can increase the risk of pathological fractures. Such conditions as hypothalamic-pituitary insufficiency (functional hypothalamic amenorrhea, anorexia nervosa, Kallmann syndrome, hyperprolactinemia), ovarian failure (gonadal dysgenesis, premature ovarian failure) and iatrogenic treatment (surgery, chemotherapy, radiotherapy) can cause hypoestrogenism. The treatment of osteopenia and osteoporosis caused by hypoestrogenism is very essential and multidirectional. The crucial role of the therapy is the achievement of proper serum estradiol concentration and eliminate the causes of hypoestrogenism.


Capacity of UV-irradiated human fibroblasts to support adenovirus DNA synthesis correlates with transcription-coupled repair and is reduced in SV40-transformed cells and cells expressing mutant p53.
We have examined the capacity of UV-irradiated human diploid fibroblasts to support adenovirus (Ad) DNA synthesis in order to assess repair of UV-damaged DNA. The capacity of UV-irradiated xeroderma pigmentosum group C (XP-C) fibroblasts to support Ad DNA synthesis was similar to that of UV-irradiated normal diploid fibroblasts, following UV exposures of greater than 9 J/m2. In contrast, XP-A, Cockayne syndrome groups A and B (CS-A and CS-B) fibroblasts were reduced in their capacity to support Ad DNA synthesis compared to normal diploid fibroblasts following a similar UV treatment. These results demonstrate that the capacity of UV-irradiated fibroblasts to support Ad DNA synthesis correlates with their ability to remove UV-induced DNA damage from active genes by transcription-coupled repair (TCR). We also demonstrate that simian virus 40 (SV40)-transformed human fibroblasts, Li-Fraumeni syndrome (LFS) fibroblasts, heterozygous for mutations in one allele of the p53 gene and immortalized LFS cell lines expressing only mutant p53 are reduced in their capacity to support Ad DNA replication following similar UV treatments. These results suggest that the capacity of UV-irradiated cells to support viral DNA synthesis involves TCR of UV-damaged DNA and is disrupted by SV40 transformation and expression of mutant p53. We propose a model in which p53-dependent TCR regulates p53 stability in response to UV.


A golgi apparatus associated with mating in Tetrahymena pyriformis.
The classical Golgi apparatus has not been observed in the several strains of Tetrahymena pyriformis examined in this laboratory at the ultrastructural level when the ciliates are grown vegetatively. However, sexually active strains, when starved for the purpose of inducing conjugation, contain stacked saccules in the oral region. When such opposite mating types are mixed for mating, the stacked saccules become swollen at their ends and vesicles appear to pinch off from them. These bodies possess the configuration of the classical Golgi apparatus of other eucells. Vesicles seem to be formed from the saccules just prior to, and toward the end of conjugation, suggesting a relationship with the mating process.


[Asking answerable questions].
Using evidence-based methods to integrate current best evidence into clinical and health care decision-making is widely acknowledged as an effective way to resolve clinical problems and promote nurses' professional abilities. While many nurses are familiar with the five steps of the evidence-based method (i.e., asking, acquiring, appraising, applying, and assessing), novices often encounter difficulties in the first step (asking) because either their questions are not adequately specific or they fail to formulate the PICO structure. This article explains specific skills needed to formulate questions and provides practical examples for clinical staff in order to provide a step-by-step approach intended to make it easier for novices to perform evidence-based nursing proficiently.


Adherence to respiratory epithelia by recombinant Escherichia coli expressing Klebsiella pneumoniae type 3 fimbrial gene products.
We examined the role of Klebsiella fimbrial types 1 and 3 in mediating adherence to human buccal and tracheal cells and to lung tissue sections. We found that clinical isolates of Klebsiella pneumoniae producing type 3 fimbriae and Escherichia coli HB101 containing a recombinant plasmid encoding expression of Klebsiella type 3 fimbriae (pFK10) demonstrated increased adherence to tracheal cells, trypsinized buccal cells, and lung tissue sections, in contrast to nonfimbriate and to type 1 fimbriate bacteria. Adherence by type 3 fimbriate bacteria was inhibited by purified type 3 fimbriae and Fab fragments derived from type 3 fimbrial-specific polyclonal immunoglobulin G. Type 3 fimbriae mediated attachment to the basolateral surface of tracheal cells and to the basal epithelial cells and the basement membrane regions of bronchial epithelia. Using an E. coli transformant (pDC17/pFK52), which expresses nonadherent P fimbrial filaments, along with the type 3 fimbrial adhesin (MrkD), we demonstrated that type 3 fimbrial attachment to respiratory cells was attributable to the MrkD adhesin subunit. Subsequent experiments demonstrated that the epithelial target of the type 3 fimbrial adhesin was most likely a peptide molecule rather than a carbohydrate. The results of this study demonstrate that, in vitro, the Klebsiella type 3 fimbrial adhesin mediates adherence to human respiratory tissue.


Creating an amateur press corps of graduate students and postdoctoral fellows to cover breaking science and improve lay-writing skills.
The Science Writing Initiative for Trainees is a science communications internship program for biomedical graduate students and postdoctoral fellows at the Medical University of South Carolina. Interns serve as an amateur press corps, writing news stories and press releases about recent high-impact research articles. Since 2016, 25 interns have written more than 100 EurekAlert! press releases that have received more than a half million views. Interns learn to explain science across the translational spectrum and to convey findings in plain language to a lay audience, serving as ambassadors for science.


Multiple early transcripts and splicing of the Autographa californica nuclear polyhedrosis virus IE-1 gene.
The immediate-early IE-1 gene of Autographa californica nuclear polyhedrosis virus was cloned, and its nucleotide sequence was determined. Sequence analysis indicated that this gene would encode a protein of 582 amino acids with a predicted molecular weight of 66,822. Analysis of IE-1 gene expression during baculovirus infection identified two transcripts. One, 1.9 kilobases (kb), was expressed at constant steady-state levels throughout infection, whereas the other, 2.1 kb, was expressed only early in infection. Analysis of IE-1 cDNA clones demonstrated that the 2.1-kb transcript contained the entire 1.9-kb transcript (exon 1) plus an additional 5' end (exon 0). Genomic Southern analysis placed the exon 0 sequences on the EcoRI B fragment, 4 kilobase pairs upstream of exon 1. Sequencing of the upstream region identified an open reading frame whose 5' end was identical to the exon 0 sequences in the cDNAs. Examination of the genomic DNA sequences around the exon-exon junction revealed sequences similar to published consensus splice acceptor and donor sequences. This is the first example of splicing of any viral transcript during baculovirus infection.


Manual laxity tests for anterior cruciate ligament injuries.
The manual laxity examination is the primary means by which clinicians evaluate ACL injuries. This paper reviews the literature and identifies the following ACL laxity tests: anterior drawer test, Lachman test. MacIntosh test, jerk test, flexion rotation drawer test, Slocum test, and the Losee test. Test technique, grading, limitations, and reliability are discussed for each test. General limitations of manual laxity tests are also presented. A review of ACL anatomy and the biomechanics of the pivot shift sign are provided to facilitate an understanding of the underlying principles of ACL laxity tests. J Orthop Sports Phys Ther 1990;11(10):474-481.


Efficient migration of complex off-line computer vision software to real-time system implementation on generic computer hardware.
This paper addresses the problem of migrating large and complex computer vision code bases that have been developed off-line, into efficient real-time implementations avoiding the need for rewriting the software, and the associated costs. Creative linking strategies based on Linux loadable kernel modules are presented to create a simultaneous realization of real-time and off-line frame rate computer vision systems from a single code base. In this approach, systemic predictability is achieved by inserting time-critical components of a user-level executable directly into the kernel as a virtual device driver. This effectively emulates a single process space model that is nonpreemptable, nonpageable, and that has direct access to a powerful set of system-level services. This overall approach is shown to provide the basis for building a predictable frame-rate vision system using commercial off-the-shelf hardware and a standard uniprocessor Linux operating system. Experiments on a frame-rate vision system designed for computer-assisted laser retinal surgery show that this method reduces the variance of observed per-frame central processing unit cycle counts by two orders of magnitude. The conclusion is that when predictable application algorithms are used, it is possible to efficiently migrate to a predictable frame-rate computer vision system.


[The effect of mandibular immobilization in fracture on the function of the salivary glands].
Studies of functional changes in the salivary glands of patients with mandibular fractures treated with intermandibular immobilization have shown that the major factors responsible for these changes were not only the absence of chewing, changed quality and quantity of food, but the duration of immobilization and viscero-visceral reactions of the body as well.


Aquaporin-4 autoantibodies in Neuromyelitis Optica: AQP4 isoform-dependent sensitivity and specificity.
Neuromyelitis Optica (NMO) is an autoimmune demyelinating disease, characterized by the presence of autoantibody (NMO-IgG) to Aquaporin-4 (AQP4). NMO-IgG identification supports NMO diagnosis and several diagnostic tests have been developed, but their sensitivity is too variable, and some assay show low sensitivity. This impairs correct diagnosis of NMO. By cell based assay (CBA) we here evaluate the efficacy of different strategies to express AQP4 in mammalian cells in terms of: a) AQP4 translation initiation signals; b) AQP4 isoforms (M1 and M23) and fluorescent tag position; c) NMO serum concentration and AQP4 degradation. Our results demonstrate that when using AQP4-M1, the nucleotide in position -3 of the AUG greatly affects the AQP4-M1/M23 protein ratio, NMO-IgG binding, and consequently test sensitivity. Test sensitivity was highest with M23 expressing cells (97.5%) and only 27.5% with AQP4-M1. The fluorescent tag added to the N-terminus of AQP4-M23 considerably affected the NMO-IgG binding, and test sensitivity, due to disruption of AQP4 suprastructures. Furthermore, sera used at high concentration resulted in AQP4 degradation which affected test sensitivity. To further evaluate the reliability of the M23 based CBA test, samples of one NMO patient collected during about 2 years clinical follow-up were tested. The results of serum titer correlated with disease activity and treatment response. In conclusion, we provide a molecular explanation for the contrasting CBA test data reported and suggest the use of M23 with a C-terminus fluorescent tag as the proper test for NMO diagnosis.


Sci-fate characterizes the dynamics of gene expression in single cells.
Gene expression programs change over time, differentiation and development, and in response to stimuli. However, nearly all techniques for profiling gene expression in single cells do not directly capture transcriptional dynamics. In the present study, we present a method for combined single-cell combinatorial indexing and messenger RNA labeling (sci-fate), which uses combinatorial cell indexing and 4-thiouridine labeling of newly synthesized mRNA to concurrently profile the whole and newly synthesized transcriptome in each of many single cells. We used sci-fate to study the cortisol response in >6,000 single cultured cells. From these data, we quantified the dynamics of the cell cycle and glucocorticoid receptor activation, and explored their intersection. Finally, we developed software to infer and analyze cell-state transitions. We anticipate that sci-fate will be broadly applicable to quantitatively characterize transcriptional dynamics in diverse systems.


Clinical, microbiological, and radiographic evaluation of sealed carious dentin after minimal intervention in primary molars.
In continuation with the ultraconservative minimal intervention approach for carious lesions treatment, lesion sterilization and tissue repair (LSTR) is a novel concept of using antibacterial drugs to disinfect dentinal, pulpal, and periapical lesions. The purpose of this study is to evaluate the clinical, radiographic, and microbiologic success rate of indirect pulp treatment (IPT) using a new technique minimal excavation and LSTR with triple antibiotic paste (TAP), for primary molars. This was randomized controlled trial design. Forty-two healthy cooperative children aged 5-10 years having deep carious lesion in primary molars were randomly assigned to receive either traditional IPT with calcium hydroxide or minimal excavation and LSTR with TAP. Follow-up was done at 6 weeks, 3, 6, and 15-18 months intervals, and treatment success or failure was determined by a combination of clinical, microbiological, and radiographic findings. Qualitative data were analyzed using Pearson's Chi-square test. Mann-Whitney U nonparametric test was used for statistically significant differences between the bacterial counts (median values and percent reduction) between the two groups and the Wilcoxon sign rank test for the intragroup evaluation of bacterial counts. LSTR with TAP was found to be as effective as traditional indirect pulp treatment (P < 0.05). Minimal excavation and LSTR with TAP can be an effective treatment methodology for the management of deep carious lesions in primary molars.


Sodium permeability of a cloned small-conductance calcium-activated potassium channel.
Small-conductance Ca2+-activated potassium channels (SK(Ca) channels) are heteromeric complexes of pore-forming main subunits and constitutively bound calmodulin. SK(Ca) channels in neuronal cells are activated by intracellular Ca2+ that increases during action potentials, and their ionic currents have been considered to underlie neuronal afterhyperpolarization. However, the ion selectivity of neuronal SK(Ca) channels has not been rigorously investigated. In this study, we determined the monovalent cation selectivity of a cloned rat SK(Ca) channel, rSK2, using heterologous expression and electrophysiological measurements. When extracellular K+ was replaced isotonically with Na+, ionic currents through rSK2 reversed at significantly more depolarized membrane potentials than the value expected for a Nernstian relationship for K+. We then determined the relative permeability of rSK2 for monovalent cations and compared them with those of the intermediate- and large-conductance Ca2+-activated K+ channels, IK(Ca) and BK(Ca) channels. The relative permeability of the rSK2 channel was determined as K+(1.0)>Rb+(0.80)>NH(4)+(0.19) approximately Cs+(0.19)>Li+(0.14)>Na+(0.12), indicating substantial permeability of small ions through the channel. Although a mutation near the selectivity filter mimicking other K+-selective channels influenced the size-selectivity for permeant ions, Na+ permeability of rSK2 channels was still retained. Since the reversal potential of endogenous SK(Ca) current is determined by Na+ permeability in a physiological ionic environment, the ion selectivity of native SK(Ca) channels should be reinvestigated and their in vivo roles may need to be restated.


Laser Doppler Velocimetry study of the effect of pure oxygen breathing on retinal blood flow.
The noninvasive Laser Doppler Velocimetry (LDV) technique was used in normal volunteers to determine retinal blood flow, F, during pure oxygen breathing at atmospheric pressure. Changes in blood flow were calculated based on Poiseuille relation, F = (pi D2/4) X Vmax, where Vmax is the red blood cell maximum or center-line velocity and D, the diameter of the vessel at the site of the LDV recordings. After five minutes of pure oxygen breathing, Vmax decreased by about 53%, vessel diameter by 12%, and retinal blood flow by about 60%. In arteries, pulsatility of red blood cell velocity (Vmax, systole/Vmax, diastole) during the cardiac cycle increased by about 10%. Decrease in red blood cell velocity was detectable within 1-1 1/2 min. From these LDV flow measurements and the changes in retinal mean circulation time (MCT) reported by other investigators, based on the relation F = v/MCT, a 45% decrease in retinal blood volume (v) during 100% O2 breathing was obtained.


[The 1996 epizootic of African swine fever in the Ivory Coast].
African swine fever was reported for the first time in the Côte d'Ivoire on 16 April 1996, in Abidjan. The epizootic resulted in the death of 135,000 pigs, equivalent to 29% of the pig population, of which 80% came from the commercial sector. By November, the outbreaks had stabilised and were then eliminated. All epidemiological evidence showed that the active form of disease ceased in September 1996. A variety of factors were responsible for limiting and then halting progression of the disease from the end of August 1996, including a new strategy which gave priority to local information, better control of the situation, depopulation of the infected area in Abidjan, the epidemiological containment of most cases within the villages, and also the assistance and direct involvement of the Food and Agriculture Organization (FAO) of the United Nations. The contribution of the FAO was decisive. It provided a diagnosis of the situation, recommended emergency measures, developed a control strategy, ensured the implementation of the control programme, set up a diagnostic laboratory unit and provided training for staff, slaughtered and destroyed any remaining pigs, set up and implemented a local information programme, developed and implemented a scheme for sentinel animals and for epidemiological surveillance, and designed, performed and analysed an epidemiological survey. From the commencement of the epizootic, two major shortcomings became apparent: inadequate dissemination of information, and a shortage of technical resources allocated for the control of African swine fever. These two shortcomings were compounded by other technical and socio-economic constraints.


Impact of Clinical Decision Support Within the Electronic Medical Record on Opioid Prescribing and Dispensing.
Introduction Clinical decision support tools support prescribers and pharmacists as they select and verify appropriate opioid regimens in efforts to combat the high variability in opioid prescribing. This study seeks to examine the impact of alerts within the electronic medical record and pharmacy system on day supply of initial opioid prescribing and dispensing. Methods This retrospective study compared a 6-month pre- and postimplementation of clinical decision support tool alerts at an integrated health care system. Data were analyzed to assess changes in the day supply of an opioid at the point of initial prescribing and dispensing based on alerts. Results The best practice alert in the electronic medical record was associated with a 27% change (p = 0.007) in prescribing by the physician, which resulted in a reduction of average day supply from 12.09 to 6.58 days. The alert in the pharmacy system was associated with a 41.3% change (p < 0.001) in dispensing, which resulted in a reduction of average day supply from 13.46 to 6.96 days. Discussion To promote judicious opioid prescribing, the best practice alert in the electronic medical record led to a statistically significant change in prescribing. To support appropriate dispensing, the alert in the pharmacy system led to a statistically significant change in dispensing. Conclusion Implementation of two clinical decision support tools that mirrored Centers for Disease Control and Prevention recommendations of prescribing less than a 7-day supply when initiating opioids resulted in a decrease in day supply of the opioid prescription for patients identified as opioid-naïve at the point of prescribing and dispensing.


[Antiviral defense by APOBEC3 family proteins].
APOBEC3G is a potent antiretroviral factor, which belongs to the APOBEC superfamily of cytidine deaminases. It deaminates cytidine to uridine in nascent minus-strand viral DNA, inducing G-to-A hypermutation in the plus-strand viral DNA. HIV-1 Vif protein overcomes the antiviral activity of APOBEC3G by targeting it for ubiquitin-dependent degradation. Recent accumulating evidences that other members of APOBEC proteins also show antiviral activity on a wide variety of viruses suggest that APOBEC family proteins play a crucial role in an antiviral defense as an innate immunity. Here, we review recent progress in research on APOBEC3 proteins.


Elena: A case of dissociative identity disorder from the 1920s.
In 1930, Italian psychiatrist Giovanni Enrico Morselli described the history, diagnosis, and treatment of his patient Elena. The case of Elena has been considered in literature as one of the most remarkable cases of multiple personality ever published. In fact, before treatment, Elena showed alternating French- and Italian-speaking personalities, with the Italian personality knowing nothing of her French counterparts. After a difficult treatment involving recovered memories of incestuous attacks by her father, which were proven to be true, Elena fully recovered from her symptoms. In this article, the author presents details of the case that were not available in the international literature before. He also discusses Elena's psychological and somatoform symptoms according to a contemporary perspective on the relationally traumatic origins of dissociation and dissociative identity disorder.


Biased genotype distributions of Candida albicans strains associated with vulvovaginal candidosis and candidal balanoposthitis in China.
Vulvovaginal candidosis (VVC), which is most frequently caused by Candida albicans, is one of the most common vaginal infections and is a common problem worldwide. Despite the fact that extensive epidemiological studies have been performed, what triggers VVC, especially recurrence of the infection, is still uncertain. Genotypes of C. albicans strains associated with VVC and balanoposthitis and of strains isolated from samples from vaginas of asymptomatic women and from various extragenital sites were determined with use of C. albicans microsatellite locus I polymorphism analysis. Genetic similarity of representative strains with the same and different C. albicans microsatellite locus I genotypes were examined by sequence analysis of housekeeping genes CaADP1, CaSYA1, and CaVPS13. The C. albicans microsatellite locus I genotypes of independent C. albicans strains isolated from samples from extragenital sites were mostly of individual specificity. In contrast, strains associated with VVC were mainly concentrated to a few genotypes, with genotypes 30-45 and 32-46 being the most common. The overall frequencies of the 2 genotypes among C. albicans strains from vaginal samples from patients with VVC and from asymptomatic women were 59.1% and 24.0%, respectively (P = .002); the frequencies among patients with complicated VVC and among patients with uncomplicated VVC were 69.2% and 35.7%, respectively (P = .003). A similar genotype distribution pattern of C. albicans strains associated with balanoposthitis was also revealed. The genetic similarity of strains with the dominant genotypes associated with both VVC and balanoposthitis was confirmed by sequence analysis of the 3 genes. The results suggest the existence of vaginopathic C. albicans strains with enhanced virulence and tropism for the vagina and the high possibility of sexual transmission of genital C. albicans infection. Identification of specific genotypes that correlate with severity of VVC is also of diagnostic and therapeutic significance.


Modelling geographically referenced survival data with a cure fraction.
The emergence of geographical information systems and related softwares nowadays enables medical databases to incorporate the geographical information on patients, allowing studies in spatial associations. Public health administrators and researchers are often interested in detecting variation in survival patterns by region or county in order to understand the possible factors that contribute towards such spatial discrepancies. These issues have led statisticians to develop survival models that account for spatial clustering and variation. Additionally, with rapid developments in medical and health sciences, researchers increasingly encounter data sets where a substantial portion of patients are cured. Models accounting for cure in the population assist in the prognosis of potentially terminal diseases. This article proposes a Bayesian modelling framework that models spatial associations for areally referenced survival data using a general class of cure models proposed by Cooner et al. The special models we outline are alternatives to the traditional proportional hazards models and can be fitted using standard Bayesian software such as WinBUGS.


Non-Invasive In Vivo Imaging and Quantification of Tumor Growth and Metastasis in Rats Using Cells Expressing Far-Red Fluorescence Protein.
Non-invasive in vivo imaging is emerging as an important tool for basic and preclinical research. Near-infrared (NIR) fluorescence dyes and probes have been used for non-invasive optical imaging since in the NIR region absorption and auto fluorescence by body tissue is low, thus permitting for greater penetration depths and high signal to noise ratio. Currently, cell tracking systems rely on labeling cells prior to injection or administering probes targeting the cell population of choice right before imaging. These approaches do not enable imaging of tumor growth, as the cell label is diluted during cell division. In this study we have developed cell lines stably expressing the far-red fluorescence protein E2-Crimson, thus enabling continuous detection and quantification of tumor growth. In a xenograft rat model, we show that E2-Crimson expressing cells can be detected over a 5 week period using optical imaging. Fluorescence intensities correlated with tumor volume and weight and allowed for a reliable and robust quantification of the entire tumor compartment. Using a novel injection regime, the seeding of MDA-MB-231 breast cancer cells in the lungs in a rat model was established and verified.


[Method of arrest and fixation of growth of yeast cultures in the postincubation period].
In order to arrest the growth and to fix yeast cultures during the postincubation period on natural and synthetic nutrient media, 10% oxalic acid can be used. The fixative is used at a concentration of 0.025 and 0.05 ml/l. Therefore, it induces on noticeable dilution of the suspension and allows precise measurements of the yeast weight.


Genome-wide CpG island profiling of intraductal papillary mucinous neoplasms of the pancreas.
Intraductal papillary mucinous neoplasms (IPMN) are precursors to infiltrating pancreatic ductal adenocarcinomas. Widespread epigenetic alterations are characteristic of many cancers, yet few studies have systematically analyzed epigenetic alterations of neoplastic precursors. Our goal was to conduct genome-wide CpG island methylation profiling to identify aberrantly methylated loci in IPMNs. We compared the CpG island methylation profiles of six IPMNs to normal primary pancreatic duct samples using methylation CpG island amplification (MCA) and Agilent CpG island microarray (MCAM) analysis. When selected 13 genes identified as differentially methylated by MCAM for methylation-specific PCR (MSP) analysis in an independent set of IPMNs and normal pancreas samples and conducted expression analysis of selected genes. We identified 2,259 loci as differentially methylated in at least one of six IPMNs including 245 genes hypermethylated in IPMNs with high-grade dysplasia compared with normal pancreatic duct samples. Eleven of 13 genes evaluated by MSP were more commonly methylated in 61 IPMNs than in 43 normal pancreas samples. Several genes (BNIP3, PTCHD2, SOX17, NXPH1, EBF3) were significantly more likely to be methylated in IPMNs with high-grade than with low-grade dysplasia. One gene, SOX17, showed loss of protein expression by immunohistochemistry in 22% (19 of 88) of IPMNs. The most specific marker, BNIP3, was not methylated in any IPMNs with low-grade dysplasia or in normal pancreas samples. IPMNs undergo extensive aberrant CpG island hypermethylation. The detection of genes selectively methylated in high-grade IPMNs such as BNIP3 may have use in the clinical evaluation of IPMNs.


Rectal temperature after marathon running.
Rectal temperature was measured in 62 male runners who competed in the 1983 Dundee marathon race: all measurements were made immediately after the race. Competitors' times were noted at 5, 10, 15 and 20 miles (8.0, 16.1, 24.1, 32.2 km) and at the finish (26.2 miles, 42.2 km). Mean finishing time of the group was 3 hr 33 min +/- 48 min (mean +/- S.D.; range = 2 hr 17 min-5 hr 11 min). Mean running speed of the group decreased progressively as the distance covered increased. Mean post-race rectal temperature was 38.7 +/- 0.9 degrees C (range 35.6-40.3 degrees C). The post-race temperature was correlated (p less than 0.01) with the time taken to cover the last 6.2 miles (10 km) of the race, but not with the overall finishing time (p greater than 0.05). Only the fastest runners were able to maintain an approximately constant pace throughout the race, whereas the slower runners slowed down progressively. The runners with the highest post-race temperature, although not necessarily the fastest runners, also tended to maintain a steady pace throughout. The runners with the lowest post-race temperature slowed down markedly only over the last 6.2 mile section of the race. The results clearly indicate that runners forced by fatigue or injury to slow down in the latter stages of races held at low ambient temperatures may already be hypothermic or at serious risk of hypothermia.


TRPV4: an exciting new target to promote alveolocapillary barrier function.
Transient receptor potential (TRP) channels are emerging as important players and drug targets in respiratory disease. Amongst the vanilloid-type TRP channels (which includes the six members of the TRPV family), target diseases include cough, asthma, cancer, and more recently, pulmonary edema associated with acute respiratory distress syndrome. Here, we critically evaluate a recent report that addresses TRPV4 as a candidate target for the management of acute lung injury that develops as a consequence of aspiration of gastric contents, or acute chlorine gas exposure. By use of two new TRPV4 inhibitors (GSK2220691 or GSK2337429A) and a trpv4(-/-) mouse strain, TRPV4 was implicated as a key mediator of pulmonary inflammation after direct chemical insult. Additionally, applied therapeutically, TRPV4 inhibitors exhibited vasculoprotective effects after chlorine gas exposure, inhibiting vascular leakage, and improving blood oxygenation. These observations underscore TRPV4 channels as candidate therapeutic targets in the management of lung injury, with the added need to balance these against the potential drawbacks of TRPV4 inhibition, such as the danger of limiting the immune response in settings of pathogen-provoked injury.


Electrocardiographic features of failed and recurrent right ventricular outflow tract catheter ablation of idiopathic ventricular arrhythmias.
Various ECG algorithms have been proposed to identify the origin of idiopathic outflow tract (OT)-ventricular arrhythmia (VA). However, electrocardiographic features of failed and recurrent right ventricular outflow tract (RVOT) ablation of idiopathic OT-VAs have not been clearly elucidated. A total of 264 consecutive patients (mean age: 44.0 ± 13.0 years, 96 male) undergoing RVOT ablation for OT-VAs with a transition ≥V3 , including 241 patients (91.6%) with initially successful procedures and 23 patients (8.4%) with failed ablation. Detailed clinical characteristics and ECG features were analyzed and compared between the two groups. VAs with failed RVOT ablation had larger peak deflection index (PDI), longer V2 R wave duration (V2 Rd), smaller V2 S wave amplitude, higher R/S ratio in V2 , higher V3 R wave amplitude, and larger V2 transition ratio than those with successful ablation. Multivariate analysis demonstrated that PDI, V2 Rd, V2 transition ratio, and pacemapping score acquired during mapping independently predicted failed ablation (P = 0.01, P = 0.01, P = 0.01, and P < 0.001, respectively). In 31 recurrent cases (12.8%) after initially successful ablation, multivariate Cox regression analysis showed that only the earliest activation time acquired during mapping predicted the recurrences after successful ablation (P = 0.001). The recurrent cases displayed different ECG features comparing with those with failed ablation. The electrocardiographic features of failed RVOT ablation of idiopathic OT-VAs with a transition ≥V3 were characterized by PDI, V2 Rd, V2 transition ratio, and pacemapping score acquired during mapping, unlike the recurrent RVOT ablation.


Speciation of oxaliplatin adducts with DNA nucleotides.
This paper describes a set of fast and selective high performance liquid chromatography (HPLC) methods coupled to electro-spray ionisation linear ion trap mass spectrometry (ESI-MS), sector-field inductively coupled plasma mass spectrometry (SF-ICP-MS) and UV detection for in vitro studies of the bifunctional adducts of oxaliplatin with mono-nucleotides, di-nucleotides and cellular DNA. The stationary phases and the optimised conditions used for each separation are discussed. Interaction of oxaliplatin with A and G mono-nucleotides resulted in the formation of five bifunctional platinum diaminocyclohexane (DACHPt) adducts. These were two isomers of the A-DACHPt-A and A-DACHPt-G adducts, and one G-DACHPt-G adduct, as confirmed by MS/MS spectra obtained by collision induced dissociation. These adducts were also characterised by UV absorption data and SF-ICP-MS elemental (195)Pt and (31)P signals. Further, interaction of oxaliplatin with AG and GG di-nucleotides resulted in the formation of three adducts: DACHPt-GG and two isomers of the DACHPt-AG adduct, as confirmed by ESI-MS and the complementary data obtained by UV and SF-ICP-MS. Finally, a very sensitive LC-ICP-MS method for the quantification of oxaliplatin GG intra-strand adducts (DACHPt-GG) was developed and used for monitoring the in vitro formation and repair of these adducts in human colorectal cancer cells. The method detection limit was 0.14 ppb Pt which was equivalent to 0.22 Pt adduct per 10(6) nucleotides based on a 10 μg DNA sample. This detection limit makes this method suitable for in vivo assessment of DACHPt-GG adducts in patients undergoing oxaliplatin chemotherapy.


Cerebral microembolus generation in different extracorporeal circulation systems.
Microemboli generated during extracorporeal circulation (ECC) are likely to induce neurological sequelae. This study examines whether the choice of a distinct type of ECC can reduce intracerebral emboli counts. Middle cerebral artery blood flow during coronary artery bypass grafting was monitored continuously by transcranial Doppler ultrasound in 45 patients. The ECC systems used were a roller pump (n = 16), a centrifugal pump (n = 18) and a combination of centrifugal pump and heparin-coated ECC system (n = 11). Patients' characteristics as well as surgical and anesthesiological procedure did not differ between the groups. Total counts did not differ significantly between the three groups. Intraoperative events in individual patients may lead to massive embolus generation overcoming positive properties of a distinct ECC system.


Molecular, biochemical and functional analysis of a novel and developmentally important fibrillar collagen (Hcol-I) in hydra.
The body wall of hydra (a member of the phylum Cnidaria) is structurally reduced to an epithelial bilayer with an intervening extracellular matrix (ECM). Previous studies have established that cell-ECM interactions are important for morphogenesis and cell differentiation in this simple metazoan. The ECM of hydra is particularly interesting because it represents a primordial form of matrix. Despite progress in our understanding of hydra ECM, we still know little about the nature of hydra collagens. In the current study we provide a molecular, biochemical and functional analysis of a hydra fibrillar collagen that has similarity to vertebrate type I and type II collagens. This fibrillar collagen has been named hydra collagen-I (Hcol-I) because of its structure and because it is the first ECM collagen to be identified in hydra. It represents a novel member of the collagen family. Similar to vertebrate type I and II collagens, Hcol-I contains an N-terminal propeptide-like domain, a triple helical domain containing typical Gly-X-Y repeats and a C-terminal propeptide domain. The overall identity to vertebrate fibrillar collagens is about 30%, while the identity of the C-terminal propeptide domain is 50%. Because the N-terminal propeptide domain is retained after post-translational processing, Hcol-I does not form thick fibers as seen in vertebrates. This was confirmed using transmission electron microscopy to study rotary shadow images of purified Hcol-I. In addition, absence of crucial lysine residues and an overall reduction in proline content, results in reduced crosslinking of fibrils and increased flexibility of the molecule, respectively. These structural changes in Hcol-I help to explain the flexible properties of hydra ECM. Immunocytochemical studies indicate that Hcol-I forms the 10 nm fibrils that comprise the majority of molecules in the central fibrous zone of hydra ECM. The central fibrous zone resides between the two subepithelial zones where hydra laminin is localized. While previous studies have shown that basal lamina components like laminin are expressed by the endoderm, in situ hybridisation studies show that Hcol-I mRNA expression is restricted to the ectoderm. Hcol-I expression is upregulated during head regeneration, and antisense studies using thio-oligonucleotides demonstrated that blocking the translation of Hcol-I leads to a reversible inhibition of head morphogenesis during this regenerative process. Taken in total, the data presented in this study indicate that Hcol-I is required for morphogensis in hydra and represents a novel fibrillar collagen whose structural characteristics help to explain the unique biophysical properties of hydra ECM. Interestingly, the structure of Hcol-I mimics what is seen in Ehlers-Danlos syndrome type VII in humans; an inherited pathological condition that leads to joint and skin abnormalities. Hcol-I therefore illustrates an adaptive trait in which the normal physiological situation in hydra translates into a pathological condition in humans.


The effectiveness of various cleaning regimens and current guidelines in contact lens case biofilm removal.
Lens case hygiene is important for safe contact lens wear. However, there are no evidence-based data to suggest optimum hygiene regimens. This in vitro study aimed to evaluate and compare the effectiveness of manufacturers' guidelines and several other regimens in removing biofilm using various types of contact lens cases and disinfecting agents. Biofilms of Staphylococcus aureus ATCC 6538 and Pseudomonas aeruginosa 122 were formed on two types of unused contact lens cases. Subsequently, each contact lens case was subjected to one of four cleaning regimens using two different multipurpose disinfecting solutions or distilled water: "rinse and air-dry (guidelines)," "rub, rinse and air-dry," "tissue-wipe and air-dry," and "rub, rinse, tissue-wipe, and air-dry." The levels of residual biofilm were then quantified using viable counts and compared. The manufacturers' guidelines resulted in 1 to 2 log CFU reduction of either biofilm. "Rub, rinse, tissue-wipe, and air-dry" was the most effective cleaning regimen (P < 0.001), capable of removing 4 to 6 log CFUs of bacteria; higher levels of biofilm were removed by mechanical friction from nonridged cases than that of ridged cases (P < 0.001). Biofilm removal varied with multipurpose solution tested. Current manufacturers' guidelines are not adequate in eliminating microbial contamination. Simply incorporating a rubbing/wiping step in daily case hygiene reduces viable organism recovery. Factors such as the cleaning regimen, antimicrobial potency of multipurpose solution, and the interior topography of a lens case may impact the surface detachment of biofilm during the cleaning process.


[Control of arterial hypertension in Spain, 1996].
Arterial Hypertension remains a serious public health problem in Spain, given its high degree of prevalence (20% to 30% of individuals suffer from hypertension, the figure increasing with age) and contributes to the main cause of death in the country, that being cardiovascular disease. 15-20% of coronary caused deaths and 20-25% of cerebral vascular deaths of Spanish adults are attributed to arterial hypertension. The level of control, despite having improved, is still low (less than 30% of patients treated). Therapeutic observation does not exceed 50%. Costs are extremely high, close to 120,000 million pesetas per annum (pharmacological treatment comprising 10-20% of the figure). Despite the progress achieved in recent years, the above mentioned figures justify the need to continue promoting and combine activities designed to deal with the problem, such as those referred to in this report. The report is directed to health professionals involved in arterial hypertension work and is intended to achieve greater levels of prevention and control of such a serious problem in Spain. A team of Spanish experts describe updated and consensual issues relating to the criteria defining arterial hypertension in adults, children and pregnant women; refractory and accelerated/malignant hypertension and serious and emergency cases of hypertension; accurate measurement of blood pressure and measuring techniques (including MAPA); diagnosis and control of hypertension (highlighting the role of Primary Medical and Hospital Services); pharmacological and non-pharmacological treatment objectives and programmes; hypertension considerations associated to special situations; as well as health education and patient compliance with therapy and bio-ethical, effectiveness, research and training issues.


Neuroprotective effects of resveratrol in an MPTP mouse model of Parkinson's-like disease: possible role of SOCS-1 in reducing pro-inflammatory responses.
In the present study we used a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) mouse model to analyze resveratrol neuroprotective effects. The MPTP-induced PD model is characterized by chronic inflammation, oxidative stress and loss of the dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc). We observed that resveratrol treatment significantly reduced glial activation, decreasing the levels of IL-1β, IL-6 and TNF-α, as well as their respective receptors in the SNpc of MPTP-treated mice, as demonstrated by Western blotting, RT-PCR and quantitative PCR analysis. This reduction is related to possible neuroprotection as we also observed that resveratrol administration limited the decline of tyrosine hydroxylase-immunoreactivity induced in the striatum and SNpc by MPTP injection. Consistent with these data, resveratrol treatment up-regulated the expression of the suppressor of cytokine signaling-1 (SOCS-1), supporting the hypothesis that resveratrol protects DA neurons of the SNpc against MPTP-induced cell loss by regulating inflammatory reactions, possibly through SOCS-1 induction.


[Expression of the surface membrane receptors of neutrophils in myeloproliferative diseases].
For 147 patients with myeloproliferative diseases, a study was made of the expression of Fc-receptors to immunoglobulins IgC, IgA, and that of FcH-receptor and receptors to C3-components of the complement in peripheral blood neutrophils. The data obtained show the lower level of neutrophils having membrane receptors in patients with acute myeloblastic leukemia and chronic granulocytic leukemia at the stage of blast transformation. A decrease in expression of membrane receptors of neutrophils was shown in patients with chronic granulocytic leukemia and benign subleukemic myelosis. The finding of a higher level of neutrophils having surface receptors revealed in patients with real polycythemia is close to the data obtained in the study of expression of membrane receptors in patients with chronic myeloproliferative diseases and healthy persons.


A phase I pharmacokinetic and pharmacodynamic study of intravenous calcitriol in combination with oral gefitinib in patients with advanced solid tumors.
In preclinical models, calcitriol and the tyrosine kinase inhibitor gefitinib are synergistic and modulate extracellular signal-regulated kinase (Erk) and Akt pathways. Therefore, we conducted a phase I study of calcitriol and gefitinib to determine the maximum tolerated dose (MTD) of this combination. Calcitriol was given i.v. over 1 h on weeks 1, 3, and weekly thereafter. Gefitinib was given at a fixed oral daily dose of 250 mg starting at week 2 (day 8). Escalation occurred in cohorts of three patients until the MTD was defined. Pharmacokinetic studies were done for calcitriol and gefitinib. Serial skin biopsies were done to investigate epidermal growth factor receptor (EGFR) pathway pharmacodynamic interactions. Thirty-two patients were treated. Dose-limiting hypercalcemia was noted in two of four patients receiving 96 mug/wk of calcitriol. One of seven patients developed dose-limiting hypercalcemia at the MTD 74 mug/wk calcitriol dose level. The relationship between calcitriol dose and peak serum calcitriol (C(max)) and systemic exposure (AUC) was linear. Mean (+/-SD) serum calcitriol C(max) at the MTD was 6.68 +/- 1.42 ng/mL. Gefitinib treatment inhibited EGFR, Akt, and Erk phosphorylation in the skin. Calcitriol did not have consistent effects on skin EGFR or its downstream elements. The combination of gefitinib and calcitriol did not modulate tumor EGFR pathway in patients with serial tumor biopsies. High doses of weekly i.v. calcitriol can be administered safely in combination with gefitinib. Calcitriol concentrations achieved at the MTD 74 mug calcitriol exceed in vivo concentrations associated with antitumor activity in preclinical models.


More than 95% reversal of left-right axis induced by right-sided hypodermic microinjection of activin into Xenopus neurula embryos.
In recent years, genes that show left-right (L-R) asymmetric expression patterns have been identified one after another in vertebrate gastrula-neurula embryos. However, we still have little information about when the irreversible L-R specification is established in vertebrate embryos. In this report, we show that almost 100% of the embryos develop to be L-R-inverted larvae after microinjection of activin molecules into the right lateral hypodermic space of Xenopus neurula embryos. After right-side injection of 10-250 pg activin protein, both early neurulae just after gastrulation movement (stage 13-14) and late neurulae just before neural tube closure (stage 17-18) showed almost 100% reversal of the heart and gut L-R axes. At higher doses of activin, more than 90% of the L-R-inverted embryos showed L-R reversal of both heart and gut. The survival ratio of the right-injected 4-day embryos was 90% on average. In the left-injected embryos, the occurrence of L-R inversion was less than 2% as observed in normal untreated siblings (1.7%). When the same amount of activin (1-50 pg) was microinjected into both sides of neurula embryos, the incidence of L-R inversion was reduced to 58%. The injection of activin along the dorsal midline in the trunk region also randomized the visceral L-R axis. Injection of activin into the right side changed normal left-handed expression of Xnr-1 to right-handed or bilateral expression. In contrast, left-handed expression of Pitx2 was switched to the right side by right activin injection. This is the first report of a method that achieves complete inversion of the visceral L-R axis by treatment of embryos at the neurula stage. Activin not only acts on the neurulae to cancel the original L-R specification up to the late neurula stage, but also rebuilds a new L-R axis whose left side coincides with the injection side. It is suggested that the left and right halves of neurulae have equal potential for L-R differentiation.


Comparative genomics of Salmonella enterica serovar Enteritidis ST-11 isolated in Uruguay reveals lineages associated with particular epidemiological traits.
Salmonella enterica serovar Enteritidis is a major cause of foodborne disease in Uruguay since 1995. We used a genomic approach to study a set of isolates from different sources and years. Whole genome phylogeny showed that most of the strains are distributed in two major lineages (E1 and E2), both belonging to MLST sequence type 11 the major ST among serovar Enteritidis. Strikingly, E2 isolates are over-represented in periods of outbreak abundance in Uruguay, while E1 span all epidemic periods. Both lineages circulate in neighbor countries at the same timescale as in Uruguay, and are present in minor numbers in distant countries. We identified allelic variants associated with each lineage. Three genes, ycdX, pduD and hsdM, have distinctive variants in E1 that may result in defective products. Another four genes (ybiO, yiaN, aas, aceA) present variants specific for the E2 lineage. Overall this work shows that S. enterica serovar Enteritidis strains circulating in Uruguay have the same phylogenetic profile than strains circulating in the region, as well as in more distant countries. Based on these results we hypothesize that the E2 lineage, which is more prevalent during epidemics, exhibits a combination of allelic variants that could be associated with its epidemic ability.


Comparative protein expression in different strains of the bloom-forming cyanobacterium Microcystis aeruginosa.
Toxin production in algal blooms presents a significant problem for the water industry. Of particular concern is microcystin, a potent hepatotoxin produced by the unicellular freshwater species Microcystis aeruginosa. In this study, the proteomes of six toxic and nontoxic strains of M. aeruginosa were analyzed to gain further knowledge in elucidating the role of microcystin production in this microorganism. This represents the first comparative proteomic study in a cyanobacterial species. A large diversity in the protein expression profiles of each strain was observed, with a significant proportion of the identified proteins appearing to be strain-specific. In total, 475 proteins were identified reproducibly and of these, 82 comprised the core proteome of M. aeruginosa. The expression of several hypothetical and unknown proteins, including four possible operons was confirmed. Surprisingly, no proteins were found to be produced only by toxic or nontoxic strains. Quantitative proteome analysis using the label-free normalized spectrum abundance factor approach revealed nine proteins that were differentially expressed between toxic and nontoxic strains. These proteins participate in carbon-nitrogen metabolism and redox balance maintenance and point to an involvement of the global nitrogen regulator NtcA in toxicity. In addition, the switching of a previously inactive toxin-producing strain to microcystin synthesis is reported.


Gene expression analysis of melanocortin system in vitiligo.
The melanocortin system in the skin coordinates pigmentation and immune response and could be implicated in the pathogenesis of vitiligo. We aimed to analyze changes in expression of genes involved in skin pigmentation (melanocortin system and enzymes involved in melanin synthesis). With quantitative RT-PCR we measured the mRNA expression levels of eight genes from the melanocortin system and two enzymes involved in melanogenesis. RNA was extracted from both lesional and non-lesional skin of vitiligo patients and in non-sun-exposed skin of healthy subjects. POMC (proopiomelanocortin) expression was lower in lesional skin compared to non-lesional skin. Expression of melanocortin receptors was increased in unaffected skin of vitiligo patients compared to healthy subjects and decreased in lesional skin compared to uninvolved skin of vitiligo patients, the differences were statistically significant in the cases of MC1R (melanocortin receptor 1) and MC4R (melanocortin receptor 4). TRP1 and DCT genes were down-regulated in lesional skin compared to non-lesional vitiligo skin or skin of healthy controls and up-regulated in uninvolved vitiligo skin compared to healthy control samples. In non-lesional skin, POMC expression was not elevated, possibly indicating that systemic influences are involved in up-regulation of MC receptor genes. Decreased expression of the analyzed genes in the lesional skin is not surprising, but statistically significant increased expression of studied genes in non-lesional skin from vitiligo patients is not described previously. In our mind, up-regulation of melanocortin system in non-lesional skin could be systemic compensation to restore normal pigmentation in lesions.


Graphene Oxide/Polyethylene Glycol-Stick for Thin Film Microextraction of β-Blockers from Human Oral Fluid by Liquid Chromatography-Tandem Mass Spectrometry.
A wooden stick coated with a novel graphene-based nanocomposite (Graphene oxide/polyethylene glycol (GO/PEG)) is introduced and investigated for its efficacy in solid phase microextraction techniques. The GO/PEG-stick was prepared and subsequently applied for the extraction of β-blockers, acebutolol, and metoprolol in human oral fluid samples, which were subsequently detected by liquid chromatography tandem mass spectrometry (LC-MS/MS). Experimental parameters affecting the extraction protocol including sample pH, extraction time, desorption time, appropriate desorption solvent, and salt addition were optimized. Method validation for the detection from oral fluid samples was performed following FDA (Food and Drug Administration) guidelines on bioanalytical method validation. Calibration curves ranging from 5.0 to 2000 nmol L-1 for acebutolol and 25.0 to 2000 nmol L-1 for metoprolol were used. The values for the coefficient of determination (R2) were found to be 0.998 and 0.996 (n = 3) for acebutolol and metoprolol, respectively. The recovery of analytes during extraction was 80.0% for acebutolol and 62.0% for metoprolol, respectively. The limit of detections (LODs) were 1.25, 8.00 nmol L-1 for acebutolol and metoprolol and the lower limit of quantifications (LLOQ) were 5.00 nmol L-1 for acebutolol and 25.0 nmol L-1 for metoprolol. Validation experiments conducted with quality control (QC) samples demonstrated method accuracy between 80.0% to 97.0% for acebutolol and from 95.0% to 109.0% for metoprolol. The inter-day precision for QC samples ranged from 3.6% to 12.9% for acebutolol and 9.5% to 11.3% for metoprolol. Additionally, the GO/PEG-stick was demonstrated to be reusable, with the same stick observed to be viable for more than 10 extractions from oral fluid samples.


Patient privacy protection using anonymous access control techniques.
The objective of this study is to develop a solution to preserve security and privacy in a healthcare environment where health-sensitive information will be accessed by many parties and stored in various distributed databases. The solution should maintain anonymous medical records and it should be able to link anonymous medical information in distributed databases into a single patient medical record with the patient identity. In this paper we present a protocol that can be used to authenticate and authorize patients to healthcare services without providing the patient identification. Healthcare service can identify the patient using separate temporary identities in each identification session and medical records are linked to these temporary identities. Temporary identities can be used to enable record linkage and reverse track real patient identity in critical medical situations. The proposed protocol provides main security and privacy services such as user anonymity, message privacy, message confidentiality, user authentication, user authorization and message replay attacks. The medical environment validates the patient at the healthcare service as a real and registered patient for the medical services. Using the proposed protocol, the patient anonymous medical records at different healthcare services can be linked into one single report and it is possible to securely reverse track anonymous patient into the real identity. The protocol protects the patient privacy with a secure anonymous authentication to healthcare services and medical record registries according to the European and the UK legislations, where the patient real identity is not disclosed with the distributed patient medical records.


Ehrlichia canis-like agent isolated from a man in Venezuela: antigenic and genetic characterization.
We report the first isolation and molecular and antigenic characterization of a human ehrlichial species in South America. A retrospective study was performed with serum specimens from 6 children with clinical signs suggestive of human ehrlichiosis and 43 apparently healthy adults who had a close contact with dogs exhibiting clinical signs compatible with canine ehrlichiosis. The evaluation was performed by the indirect fluorescent-antibody assay with Ehrlichia chaffeensis Arkansas, Ehrlichia canis Oklahoma, and Ehrlichia muris antigens. The sera from two apparently healthy humans were positive by the indirect fluorescent-antibody assay for all three antigens. Of the three antigens, samples from humans 1 and 2 showed the highest antibody titers against E. chaffeensis and E. muris, respectively. The remaining serum samples were negative for all three antigens. One year later examination of a blood sample from subject 1 revealed morulae morphologically resembling either E. canis, E. chaffeensis, or E. muris in monocytes in the blood smear. The microorganism, referred to here as Venezuelan human ehrlichia (VHE), was isolated from the blood of this person at 4 days after coculturing isolated blood leukocytes with a dog macrophage cell line (DH82). The organism was also isolated from mice 10 days after intraperitoneal inoculation of blood leukocytes from subject 1. Analysis by electron microscopy showed that the human isolate was ultrastructurally similar to E. canis, E. chaffeensis, and E. muris. When the virulence of VHE in mice was compared with those of E. chaffeensis, E. canis, and E. muris, only VHE and E. muris induced clinical signs in BALB/c mice at 4 and 10 days, respectively, after intraperitoneal inoculation. VHE was reisolated from peritoneal exudate cells of the mice. Only E. chaffeensis- and E. muris-infected mice developed significant splenomegaly. Western immunoblot analysis showed that serum from subject 1 reacted with major proteins of the VHE antigen of 110, 80, 76, 58, 43, 35, and 34 kDa. Human serum against E. chaffeensis reacted strongly with 58-, 54-, 52-, and 40-kDa proteins of the VHE antigen. Anti-E. canis dog serum reacted strongly with 26- and 24-kDa proteins of VHE. In contrast, anti-E. sennetsu rabbit and anti-E. muris mouse sera did not react with the VHE antigen. Serum from subject 1 reacted with major proteins of 90, 64, or 47 kDa of the E. chaffeensis, E. canis, and E. muris antigens. This reaction pattern suggests that this serum sample was similar to serum samples from E. chaffeensis-infected human patients in Oklahoma. The base sequence of the 16S rRNA gene of VHE was most closely related to that of E. canis Oklahoma. On the basis of these observations, we suggest that VHE is a new strain or a subspecies of E. canis which may cause asymptomatic persistent infection in humans.


Broth cultures yield vs traditional approach in the workup of infectious keratitis.
To elucidate whether BACTEC Peds Plus F broth, usually used for culturing body fluids in paediatric departments, can be used for corneal cultures from cases with clinically suspected infectious keratitis, and to compare yields between this method and traditional methods (blood agar, chocolate agar, a fungal media, and swab transport media). All cases with newly diagnosed, nonviral, clinically suspected infectious keratitis with no prior antibiotic therapy, were cultured both in the BACTEC Peds Plus F broth and the traditional method. McNemar's test was used for pairwise comparisons of the rates of positive growth between the two groups. In total, 30 eyes were included in this study. The growth rates for the traditional method and the BACTEC broth were similar (50.0 and 53.33%, respectively, P=1.0). The overall growth rate for the two methods combined was 73.33%, which is 45.29% higher than the reported yield in the literature (average of 50.47%). Our results show that BACTEC Peds Plus F broth can be used successfully in the work-up of clinically suspected infectious keratitis. The method has, apparently, several advantages over the 'Traditional method:' time-savings, as only one medium needs to be inoculated, transportation to the laboratory is simpler as there is no need for immediate incubation, and there is no need to keep and maintain a supply of fresh agar media. This method is especially suitable for office settings and remote clinics, but also can be used in hospital setting, as an adjunct, when available, to increase the growth yield.


Order and disorder in the local and long-range structure of the spin-glass pyrochlore, Tb(2)Mo(2)O(7).
To understand the origin of the spin-glass state in molybdate pyrochlores, the structure of Tb(2)Mo(2)O(7) is investigated using two techniques: the long-range lattice structure was measured using neutron powder diffraction, and local structure information was obtained from the extended x-ray absorption fine structure technique. While the long-range structure appears generally well ordered, enhanced mean-squared site displacements on the O(1) site and the lack of temperature dependence of the strongly anisotropic displacement parameters for both the Mo and O(1) sites indicate some disorder exists. Likewise, the local structure measurements indicate some Mo-Mo and Tb-O(1) nearest-neighbor disorder exists, similar to that found in the related spin-glass pyrochlore, Y(2)Mo(2)O(7). Although the freezing temperature in Tb(2)Mo(2)O(7), 25 K, is slightly higher than in Y(2)Mo(2)O(7), 22 K, the degree of local pair distance disorder is actually less in Tb(2)Mo(2)O(7). This apparent contradiction is considered in light of the interactions involved in the freezing process.


Activators of cyclic adenosine 3':5'-monophosphate accumulation in rat hippocampal slices: action of vasoactive intestinal peptide (VIP).
The present experiments tested the ability of putative neurotransmitters and neuromodulators to regulate cyclic adenosine 3':5'-monophosphate (cAMP) levels in rat hippocampal slices. Slices from ovariectomized adult female rats were equilibrated for 1 hr and incubated for 20 min with various test compounds, and cAMP was extracted and quantified using a competitive protein-binding assay. Norepinephrine, adenosine, histamine, and prostaglandins E1 and E2 alpha, induced moderate (1.5- to 5-fold) increases in cellular cAMP, whereas dopamine, serotonin, prostaglandin F2 alpha, and glutamate were relatively ineffective. Most striking was the observation that vasoactive intestinal peptide (VIP) produced marked elevation (approximately 80-fold at 6 microM) of hippocampal slice cAMP content. In contrast, other peptides produced only 2-fold increased (glucagon, somatostatin) or no change in cellular cAMP levels (enkephalins, LHRH, ACTH analogue, arginine vasopressin). Significant elevations in cAMP were seen with VIP concentrations as low as 20 nM; the cAMP response was half-maximal at 1 microM VIP and maximized between 10 and 20 microM. At maximally effective concentrations, VIP was 86% as effective in increasing cAMP as maximal concentrations of forskolin, a compound which activates adenylate cyclase in most cell types. The cAMP response to 10 microM VIP was pronounced after a 1-min incubation (16-fold elevations) and was maximal at 30 min (140-fold elevation). When slices from other brain areas were compared, it was found that regions known to contain high levels of VIP (cerebral cortex) also responded to VIP treatment with 30- to 50-fold elevations in cAMP.(ABSTRACT TRUNCATED AT 400 WORDS)


Itsy bitsy spider: Fear and avoidance (generalization) in a free-exploratory virtual reality paradigm.
Most experimental avoidance paradigms lack either control over the experimental situation or simplify real-life avoidance behavior to a great extent, making it difficult to generalize the results to the complex approach-avoidance situations that anxious individuals face in daily life. The current study aimed to examine the usability of our recently developed free-exploratory avoidance paradigm in Virtual Reality (VR) that allows for the assessment of subjective as well as behavioral avoidance in participants with varying levels of spider fear. In a VR escape room, participants searched for cues to decipher a code-locked door. Opening a wooden box marked with a post-it note (conditioned stimulus, CS) resulted in exposure to a virtual crawling spider (unconditioned stimulus, US). Avoidance of the original CS and other objects marked with the CS (generalization stimuli, GSs; EXPgen condition) or non-marked (CONT condition) objects was measured via questionnaires and relative manipulation times in a novel room. We expected a positive linear relationship between US aversiveness (levels of spider fear) and (generalization of) fear and avoidance behaviors. Avoidance learning and generalization was demonstrated on both a subjective and behavioral level. Higher levels of spider fear were, overall, related to more negative emotions in response to the encounter with the spider, higher US expectancies for the GSs, and more self-reported and behavioral avoidance of the original CS and the GSs. Finally, we explored relationships between trait anxiety and intolerance of uncertainty and fear and avoidance (generalization), but no robust associations were observed. In conclusion, we confirmed the expected positive linear relationship between spider fear and (generalization of) fear and avoidance behaviors. Our results suggest that our free-exploratory VR avoidance paradigm is well-suited to investigate avoidance behaviors and the generalization of avoidance.


Relationship between phospholipase C-zeta, semen parameters, and chromatin status.
The need for additional tests to complement basic sperm analysis in clinics is well appreciated. In this regard, a number of tests such as sperm DNA integrity test as a tool in diagnosis and treatment of infertility are suggested. But recent studies have focused on main sperm factors involved in oocyte activation such as phospholipase C-zeta (PLCζ) that initiate intracellular Ca2+ signaling and embryogenesis. Therefore, this study aimed to investigate the relationship between PLCζ, basic semen parameters, sperm DNA fragmentation (SDF), and protamine deficiency in men with normal (n=32) and abnormal (n=23) semen parameters. Unlike SDF and protamine deficiency, as negative factors related to fertility, the mean value of PLCζ as positive factor related to infertility was significantly lower in men with abnormal semen parameters compared to men with normal semen parameters. Significant correlations were also observed between sperm concentration, motility, and abnormal morphology with the percentage of PLCζ positive spermatozoa. In addition, logistic regression analysis revealed that sperm morphology is more predictive than sperm motility and concentration for PLCζ presence. In addition, a statistically significant negative relationship was observed between the percentage of PLCζ positive spermatozoa and SDF. These findings suggested during ICSI, selection of sperm based on morphology has a profound effect on its ability to induce oocyte activation based on the likelihood of PLCζ expression. Therefore, assessment of PLCζ as an index for fertilization potential of a semen sample in men with severe teratozoospermia may define individuals who are candidates for artificial oocyte activation (AOA) and may avoid failed fertilization post ICSI.


Ethanolic Extract of Bark from Salix aegyptiaca Ameliorates 1,2-dimethylhydrazine-induced Colon Carcinogenesis in Mice by Reducing Oxidative Stress.
We have previously shown that ethanolic extract from bark (EEB) of Salix aegyptiaca (Musk Willow) can inhibit proliferation and motility and induce apoptosis in colon cancer cells. Tandem mass spectrometry revealed EEB to be rich in catechin, catechol, and salicin. The present study investigated the chemopreventive effect of HPLC-fingerprinted EEB on 1,2-dimethylhydrazine (DMH)-induced aberrant crypt foci (ACF) formation in mice. DMH (20 mg/kg body weight) was weekly injected subcutaneously to mice for the first 2 weeks. EEB (100 and 400 mg/kg body weight) was provided orally from the 7th to 14th week, after which colon tissues were evaluated histologically and biochemically. DMH treatment induced high number of ACF; EEB significantly reduced the number and multiplicity of ACF, along with a restoration in goblet cells and mucin accumulation. EEB supplementation improved the markers of inflammation (myeloperoxidase and neutrophil infiltration) and oxidative stress. More importantly, EEB amplified apoptosis of neoplastic cells in the colon mucosa of DMH-treated mice. It also lowered levels of markers for early transformation events such as EGFR, nuclear β-catenin, and COX-2 in colon cancer cell lines HT-29 and HCT-116. The innocuity of EEB (up to 1600 mg/kg) to mice reinforces its potential as a chemopreventive agent.


[Efficacy analysis of unrelated donor hematopoietic stem cell transplantation for treatment of high risk acute myeloid leukemia].
This study was purposed to investigate the therapeutic efficacy of unrelated donor hematopoietic stem cell transplantation (URD-HSCT) for patients with high risk and refractory acute myeloid leukemia (AML). Twenty-two patients with high-risk and refractory AML receive URD-HSCT were enrolled in this study. All the patients received myeloablative preconditioning regimen consisting of busulfan/cyclophosphamide (for 20 cases) or total body irradiation/cyclophosphamide (for 2 cases) before URD-HSCT. The cyclosporin A (CsA)/MTX/MMF/ATG were used to prevent the acute graft versus host disease (aGVHD). The results showed that 21 out of 22 patients acquired engraftment with implantation rate 95.5%. The median time of ANC ≥ 0.5×10(9)/L was 12 (10-19) days, and that of Plt ≥ 20×10(9)/L was 14 (5 - 22) days. The median follow-up time post transplantation was 18 (3 to 135.5) months. The 2-year overall survival (OS) and leukemia-free survival (LFS) were (53.9 ± 12.2) % and (49.1 ± 10.7)% respectively. Eight cases developed aGVHD. The cumulative incidence of aGVHD was (39.1 ± 10.6) %. Six patients developed I-II grade of aGVHD and two patients developed III-IV grade of aGVHD. The chronic graft versus host disease (cGVHD) was occurred in 6 patients (4 patients limited, 2 patients extensive) of the 19 evaluable patients. The cumulative incidence was (28.8 ± 9.6)%. Seven cases relapsed, and the cumulative response rate of 2 years was (35.8 ± 11) %. One of 9 patients died from sepsis before hematopoietic reconstruction, one died from lung infection, Six died from relapse and one relapsed patient died from IV grade of aGVHD post chemotherapy and donor lymphocyte infusion (DLI). The univariate analysis revealed that relapse was the major factor for the OS, and the sex, age, preconditioning regimen, aGVHD and infection didn't significantly influence the efficacy of URD-HSCT. The survival of patients with cGVHD was superior to those who didn't have cGVHD (83.3% vs 37%, P = 0.152). It is concluded that URD-HSCT is a safe and effective therapy for high-risk AML patients without related donor. Notably, patients with cGVHD had a better survival. Relapse is an unfavourable factor for the efficacy of URD-HSCT and adoptive immunotherapy such as DLI can prevent it and improve the prognosis to achieve the long-time survival.


MUSIC imaging applied to total internal reflection tomography.
This paper applies the multiple signal classification (MUSIC) imaging method to determine the locations of a collection of small anisotropic spherical scatterers in the framework of total internal reflection tomography. Multiple scattering between scatterers is considered, and the inverse scattering problem is nonlinear, which, however, is solved by the proposed fast analytical approach where no associated forward problem is iteratively evaluated. The paper also discusses the role of propagating and evanescent waves, the polarization of incidence waves, separation of scatterers from the surface of the substrate, and the level of noise on the resolution of imaging.


Get real: what will draw physicians to the Web?
What are physicians waiting for? What will it take to stimulate widespread adoption of Internet medical systems? How can health care leaders and physicians help the technology innovators and the executives of technology firms understand the components necessary to assure physician acceptance and utilization of new tools? (1) Don't underestimate the personal nature of a physician's practice. It really isn't a "business." (2) Most physicians are not Luddites; they are just extremely pragmatic and practical. (3) For the majority of physicians to adopt a new technology in their private office practice, it must address three major issues: money, hassle, and patient care. There are many obstacles to adopting the new technologies that are the result of physician training and expectations and the current models of payment and revenue generation. Some technological innovations are presented to physicians without sufficient respect for their knowledge of how medical practices really work. The benefits promised often don't match with the needs structure of the physicians. As a consequence, the cycle of diffusion of these new systems is extended and delayed.


Three-dimensional structure of Theiler murine encephalomyelitis virus (BeAn strain).
Depending on the strain, Theiler murine encephalomyelitis virus (TMEV) may cause acute encephalitis or chronic demyelinating disease, which is associated with viral persistence in mice. Persistent central nervous system infection and demyelination by the less-virulent TMEV has provided a useful animal model for the human demyelinating disease multiple sclerosis. The less-virulent BeAn strain of TMEV was crystallized and its atomic structure was determined by x-ray crystallography. The alpha-carbon coordinates of the closely related Mengo virus were used to calculate the initial phases to 3.5 A resolution and the interpretable electron density map was produced by 10 cycles of 30-fold noncrystallographic molecular replacement averaging. The structure revealed a high degree of overall structural similarity to Mengo virus as well as substantial differences in the surface loops. These structural changes might be correlated with TMEV host-specific recognition, pH-related stability, and neurovirulence.


DYRK1A promotes viral entry of highly pathogenic human coronaviruses in a kinase-independent manner.
Identifying host genes essential for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has the potential to reveal novel drug targets and further our understanding of Coronavirus Disease 2019 (COVID-19). We previously performed a genome-wide CRISPR/Cas9 screen to identify proviral host factors for highly pathogenic human coronaviruses. Few host factors were required by diverse coronaviruses across multiple cell types, but DYRK1A was one such exception. Although its role in coronavirus infection was previously undescribed, DYRK1A encodes Dual Specificity Tyrosine Phosphorylation Regulated Kinase 1A and is known to regulate cell proliferation and neuronal development. Here, we demonstrate that DYRK1A regulates ACE2 and DPP4 transcription independent of its catalytic kinase function to support SARS-CoV, SARS-CoV-2, and Middle East Respiratory Syndrome Coronavirus (MERS-CoV) entry. We show that DYRK1A promotes DNA accessibility at the ACE2 promoter and a putative distal enhancer, facilitating transcription and gene expression. Finally, we validate that the proviral activity of DYRK1A is conserved across species using cells of nonhuman primate and human origin. In summary, we report that DYRK1A is a novel regulator of ACE2 and DPP4 expression that may dictate susceptibility to multiple highly pathogenic human coronaviruses.


Psoriasis upregulated phorbolin-1 shares structural but not functional similarity to the mRNA-editing protein apobec-1.
Earlier studies of psoriatic and normal primary keratinocytes treated with phorbol 12-myristate-1-acetate identified two low-molecular-weight proteins, termed phorbolin-1 (20 kDa; pI 6.6) and phorbolin-2 (17.6 kDa; pI 6.5). As a first step towards elucidating the role of these proteins in psoriasis, we report here the molecular cloning and chromosomal mapping of phorbolin-1 and a related cDNA that codes for a protein exhibiting a similar amino acid sequence. The phorbolins were mapped to position 22q13 immediately centromeric to the c-sis proto-oncogene. Transient expression of the phorbolin-1 cDNA in COS cells and by in vitro transcription/translation, yielded polypeptides that comigrated with phorbolins-1 and -2. Comparative sequence analysis revealed 22% overall identity and a similarity of 44% of the phorbolins to apobec-1, the catalytic subunit of the mammalian apolipoprotein B mRNA editing enzyme; however, recombinant-expressed phorbolin-1 exhibited no cytidine deaminase activity, using either a monomeric nucleoside or apolipoprotein B cRNA as substrate, and failed to bind an AU-rich RNA template. Whereas the precise function of the phorbolins remains to be elucidated, the current data suggest that it is unlikely to include a role in the post-transcriptional modification of RNA in a manner analogous to that described for apobec-1.


Mammalian Wnt3a is released on lipoprotein particles.
Little is known about the release and intercellular transport of Wnt proteins from mammalian cells. Lipoproteins may act as carriers for the intercellular movement and gradient formation of the lipid-linked morphogens Wingless and Hedgehog in Drosophila. To investigate whether such a mechanism can occur in mammals, we have studied Wnt release in cultured mammalian cells. Wnt3a associated with lipoproteins in the culture medium and not with extracellular vesicles or exosomes. Although Wnt3a was associated with both high-density lipoproteins (HDL) and low-density lipoproteins, only HDL allowed Wnt3a release from mouse fibroblasts. Remarkably, Wnt3a lacking its palmitate moiety was released in a lipoprotein-independent manner, demonstrating the dual role of palmitoylation in membrane and lipoprotein binding. We additionally found that Wnt3a can be released from enterocyte cell lines on endogenously expressed lipoproteins. We further discuss the physiological implications of our findings.


What Doctors Want: A Comment on the Financial Preferences of Organized Medicine.
Organized medicine's persistent demand for high payments is one factor that contributes to the rising costs of health care. The profession's long-standing preference for private and fee-for-service practice has pressured payers to increase reimbursement rates in fee-based systems; and it has stalled, thwarted, or otherwise co-opted attempts to contain costs in other payment systems. Yet what doctors want in fact varies. This article revisits classic comparative studies of organized medicine in advanced democracies to highlight two underemphasized findings: (1) physicians' financial preferences can deviate from traditional expectations, and (2) the structure of the organizations that represent doctors can shape whether and how those preferences are expressed. These findings remain relevant today as a discussion of contemporary American health politics illustrates.


Application of the attribution-value model of prejudice to homosexuality.
According to the attribution-value model, prejudice toward a group stems from 2 interrelated variables: attributions of controllability and cultural value. Thus, prejudice toward gay men and lesbians may stem from others' holding them responsible for their behaviors and perceiving negative cultural value regarding homosexuality. The author tested that model by using the issue of homosexuality. The participants were Turkish undergraduates who completed a homophobia scale and answered questions about the origins of homosexuality, cultural attitudes toward homosexuality, and their own gender and sexual preferences. In general, the participants were prejudiced against gay men and lesbians. As expected, attributions of controllability and negative cultural value regarding homosexuality operated jointly to explain 39% of the variation in homophobia. The participants who thought that homosexuality was controllable had more negative attitudes toward gay men and lesbians than did those who thought that homosexuality was uncontrollable. The female participants were more tolerant of homosexuality than were the male participants.


A progress report on the University Group Diabetes Program.
The increase in cardiovascularortality associated with the use of oral hypoglycemic agents is emphasized. It is important that clinical pharmacologists take a more positive stand against the use of medications that have added risks that are not offset by actual benefits to the patient.


A dynamic human health risk assessment system.
An online human health risk assessment system (OHHRAS) has been designed and developed in the form of a prototype database-driven system and made available for the population of India through a website - www.healthriskindia.in. OHHRAS provide the three utilities, that is, health survey, health status, and bio-calculators. The first utility health survey is functional on the basis of database being developed dynamically and gives the desired output to the user on the basis of input criteria entered into the system; the second utility health status is providing the output on the basis of dynamic questionnaire and ticked (selected) answers and generates the health status reports based on multiple matches set as per advise of medical experts and the third utility bio-calculators are very useful for the scientists/researchers as online statistical analysis tool that gives more accuracy and save the time of user. The whole system and database-driven website has been designed and developed by using the software (mainly are PHP, My-SQL, Deamweaver, C++ etc.) and made available publically through a database-driven website (www.healthriskindia.in), which are very useful for researchers, academia, students, and general masses of all sectors.


High efficiency, broadband solar cell architectures based on arrays of volumetrically distributed narrowband photovoltaic fibers.
We propose a novel solar cell architecture consisting of multiple fiber-based photovoltaic (PV) cells. Each PV fiber element is designed to maximize the power conversion efficiency within a narrow band of the incident solar spectrum, while reflecting other spectral components through the use of optical microcavity effects and distributed Bragg reflector (DBR) coatings. Combining PV fibers with complementary absorption and reflection characteristics into volume-filling arrays enables spectrally tuned modules having an effective dispersion element intrinsic to the architecture, resulting in high external quantum efficiency over the incident spectrum. While this new reflective tandem architecture is not limited to one particular material system, here we apply the concept to organic PV (OPV) cells that use a metal-organic-metal-dielectric layer structure, and calculate the expected performance of such arrays. Using realistic material properties for organic absorbers, transport layers, metallic electrodes, and DBR coatings, 17% power conversion efficiency can be reached.


Erythematous capillary-lymphatic malformations mimicking blood vascular anomalies.
Superficial erythematous cutaneous vascular malformations are assumed to be blood vascular in origin, but cutaneous lymphatic malformations can contain blood and appear red. Management may be different and so an accurate diagnosis is important. Cutaneous malformations were investigated through 2D histology and 3D whole-mount histology. Two lesions were clinically considered as port-wine birthmarks and another 3 lesions as erythematous telangiectasias. The aims were (i) to demonstrate that cutaneous erythematous malformations including telangiectasia can represent a lymphatic phenotype, (ii) to determine if lesions represent expanded but otherwise normal or malformed lymphatics, and (iii) to determine if the presence of erythrocytes explained the red color. Microscopy revealed all lesions as lymphatic structures. Port-wine birthmarks proved to be cystic lesions, with nonuniform lymphatic marker expression and a disconnected lymphatic network suggesting a lymphatic malformation. Erythematous telangiectasias represented expanded but nonmalformed lymphatics. Blood within lymphatics appeared to explain the color. Blood-lymphatic shunts could be detected in the erythematous telangiectasia. In conclusion, erythematous cutaneous capillary lesions may be lymphatic in origin but clinically indistinguishable from blood vascular malformations. Biopsy is advised for correct phenotyping and management. Erythrocytes are the likely explanation for color accessing lymphatics through lympho-venous shunts.