Mental fatigue in central-field and peripheral-field steady-state visually evoked potential and its effects on event-related potential responses.
The steady-state visually evoked potential (SSVEP) is a natural response of the brain to visual stimulation at specific frequencies and is used widely for electroencephalography-based brain-computer interface (BCI) systems. Although the SSVEP is useful for its high level of decoding accuracy, visual fatigue from the repetitive visual flickering is an unavoidable problem. In addition, hybrid BCI systems that combine the SSVEP with the event-related potential (ERP) have been proposed recently. These hybrid BCI systems would improve the decoding accuracy; however, the competing effect by simultaneous presentation of the visual stimulus could possibly supervene the signal in the hybrid system. Nevertheless, previous studies have not sufficiently reported these problems of visual fatigue with SSVEP stimuli or the competing effect in the SSVEP+ERP system. In this study, two different experiments were designed to explore our claims. The first experiment evaluated the visual fatigue level and decoding accuracy for the different types of SSVEP stimuli, which were the peripheral-field SSVEP (pSSVEP) and the central-field SSVEP (cSSVEP). We report that the pSSVEP could reduce the visual fatigue level by avoiding direct exposure of the eye-retina to the flickering visual stimulus, while also delivering a decoding accuracy comparable to that of cSSVEP. The second experiment was designed to examine the competing effect of the SSVEP stimuli on ERP performance and vice versa. To do this, the visual stimuli of ERP and SSVEP were presented simultaneously as part of the BCI speller layout. We found a clear competing effect wherein the evoked brain potentials were influenced by the SSVEP stimulus and the band power at the target frequencies was also decreased significantly by the ERP stimuli. Nevertheless, these competing effects did not lead to a significant loss in decoding accuracy; their features preserved sufficient information for discriminating a target class. Our work is the first to evaluate the visual fatigue and competing effect together, which should be considered when designing BCI applications. Furthermore, our findings suggest that the pSSVEP is a viable substitution for the cSSVEP because of its ability to reduce the level of visual fatigue while maintaining a minimal loss of decoding accuracy.


[National health research systems in Latin America: a 14-country review].
This article discusses the main features of the national health research systems (NHRS) of Argentina, Bolivia, Brazil, Chile, Costa Rica, Cuba, Ecuador, El Salvador, Honduras, Panama, Paraguay, Peru, Uruguay, and Venezuela, based on documents prepared by their country experts who participated in the First Latin American Conference on Research and Innovation for Health held in April 2008, in Rio de Janeiro, Brazil. The review also includes sources cited in the reports, published scientific papers, and expert opinion, as well as regional secondary sources. Six countries reported having formal entities for health research governance and management: Brazil and Costa Rica's entities are led by their ministries of health; while Argentina, Cuba, Ecuador, and Venezuela have entities shared by their ministries of health and ministries of science and technology. Brazil and Ecuador each reported having a comprehensive national policy devoted specifically to health science, technology, and innovation. Argentina, Brazil, Costa Rica, Cuba, Ecuador, Panama, Paraguay, Peru, and Venezuela reported having established health research priorities. In conclusion, encouraging progress has been made, despite the structural and functional heterogeneity of the study countries' NHRS and their disparate levels of development. Instituting good NHRS governance/management is of utmost importance to how efficiently ministries of health, other government players, and society-at-large can tackle health research.


Induction of endogenous murine type C virus by ultraviolet-irradiated herpes simplex virus: effect of metabolic inhibitors.
Ultraviolet-irradiated herpes simplex virus (u.v.-HSV) induced endogenous xenotropic type C virus from AI--2 cells, derived from the BALB/c mouse, as determined by infectious centre focus-forming assay on permissive normal rat kidney (NRK) cells. The number of cells induced to release type C virus by irradiated HSV was dependent on the level of u.v. exposure received by the HSV. Optimal induction occurred when cells were infected with irradiated HSV during their exponential growth phase. Virus induction decreased under conditions of simultaneous cellular exposure to hydroxyurea or actinomycin D, inhibitors of DNA and RNA synthesis, respectively, with actinomycin D having a greater inhibitory effect. This suggests that both DNA and RNA synthesis are required for irradiated HSV induction of murine xenotropic virus. Hydroxyurea decreased induction in the first few hours after infection of A1--2 cells with irradiated HSV, suggesting that the biological events involving DNA synthesis which are required for induction by u.v.-HSV occur shortly after infection.


Covalent Grafting of Polyoxometalate Hybrids onto Flat Silicon/Silicon Oxide: Insights from POMs Layers on Oxides.
Immobilization of polyoxometalates (POMs) onto oxides is relevant to many applications in the fields of catalysis, energy conversion/storage, or molecular electronics. Optimization and understanding the molecule/oxide interface is crucial to rationally improve the performance of the final molecular materials. We herein describe the synthesis and covalent grafting of POM hybrids with remote carboxylic acid functions onto flat Si/SiO2 substrates. Special attention has been paid to the characterization of the molecular layer and to the description of the POM anchoring mode at the oxide interface through the use of various characterization techniques, including ellipsometry, AFM, XPS, and FTIR. Finally, electron transport properties were probed in a vertical junction configuration and energy level diagrams have been drawn and discussed in relation with the POM molecular electronic features inferred from cyclic-voltammetry, UV-visible absorption spectra, and theoretical calculations. The electronic properties of these POM-based molecular junctions are driven by the POM LUMO (d-orbitals) whatever the nature of the tether or the anchoring group.


Staging of papillary thyroid carcinoma with ultrasonography: performance in a large series.
This study was designed to evaluate the performance of ultrasonography (US) for the preoperative staging of papillary thyroid carcinoma (PTC) in a large series according to the TNM classification. Preoperative US was performed for the evaluation of primary tumors and lymph node metastasis in 722 consecutive patients with cytologically proven PTC. Three experienced radiologists prospectively determined T and N categories of PTC. N categorization was based on compartments, including left and right lateral compartments (levels II-V) and central compartment (level VI). All patients underwent surgery and central compartment dissection. Lateral compartment dissection was selectively performed. We assessed the diagnostic performance of preoperative US for staging of PTC on the basis of pathologic findings of surgical specimens. Subgroup analysis according to suspicion of diffuse thyroid disease (DTD) on US was performed to compare US accuracies for N categorization. US predicted 61.7% (142/230) of patients with multifocal PTC and 67.1% (100/149) of patients with bilateral malignancy. Overall accuracy of US for T categorization was 69.7% (503/722) and that of US for N categorization was 59% (426/722). Accuracies of sonographic categorization for N0, N1a, and N1b were 66% (276/418), 33.3% (70/210), and 85.1% (80/94), respectively. Overall US accuracy for prediction of an N category was significantly lower in patients with US-indicated DTD (51.1%, 67/131) than it was in patients without DTD (60.7%, 359/591; P = 0.043). Preoperative US is a feasible technique for the preoperative staging of PTC and is helpful for the detection of lateral compartment metastasis. Presence of DTD can affect the staging of lymph node metastasis.


Proteomic and transcriptomic analyses of fecundity in the brown planthopper Nilaparvata lugens (Stål).
As an r-strategy insect species, the brown planthopper (BPH) Nilaparvata lugens (Stål) is a serious pest of rice crops in the temperate and tropical regions of Asia and Australia, which may be due to its robust fecundity. Here we combined 2-DE comparative proteomic and RNA-seq transcriptomic analyses to identify fecundity-related proteins and genes. Using high- and low-fecundity populations as sample groups, a total of 54 and 75 proteins were significantly altered in the third and sixth day brachypterous female stages, respectively, and 39 and 54 of these proteins were identified by MALDI-TOF/TOF MS. In addition, 71,966 unigenes were quantified by Illumina sequencing. On the basis of the transcriptomic analysis, 7408 and 1639 unigenes demonstrated higher expression levels in the high-fecundity population in the second day brachypterous female adults and the second day fifth instar nymphs, respectively, and 411 unigenes were up-regulated in both groups. Of these dozens of proteins and thousands of unigenes, five were differentially expressed at both the protein and mRNA levels at all four time points, suggesting that these genes may regulate fecundity. Glutamine synthetase (GS) was chosen for further functional studies. RNAi knockdown of the GS gene reduced the fecundity of N. lugens by 64.6%, disrupted ovary development, and inhibited vitellogenin (Vg) expression. Our results show that a combination of proteomic and transcriptomic analyses provided five candidate proteins and genes for further study. The knowledge gained from this study may lead to a more fundamental understanding of the fecundity of this important agricultural insect pest.


Genetic deficiency of glycogen synthase kinase-3beta corrects diabetes in mouse models of insulin resistance.
Despite treatment with agents that enhance beta-cell function and insulin action, reduction in beta-cell mass is relentless in patients with insulin resistance and type 2 diabetes mellitus. Insulin resistance is characterized by impaired signaling through the insulin/insulin receptor/insulin receptor substrate/PI-3K/Akt pathway, leading to elevation of negatively regulated substrates such as glycogen synthase kinase-3beta (Gsk-3beta). When elevated, this enzyme has antiproliferative and proapoptotic properties. In these studies, we designed experiments to determine the contribution of Gsk-3beta to regulation of beta-cell mass in two mouse models of insulin resistance. Mice lacking one allele of the insulin receptor (Ir+/-) exhibit insulin resistance and a doubling of beta-cell mass. Crossing these mice with those having haploinsufficiency for Gsk-3beta (Gsk-3beta+/-) reduced insulin resistance by augmenting whole-body glucose disposal, and significantly reduced beta-cell mass. In the second model, mice missing two alleles of the insulin receptor substrate 2 (Irs2-/-), like the Ir+/- mice, are insulin resistant, but develop profound beta-cell loss, resulting in early diabetes. We found that islets from these mice had a 4-fold elevation of Gsk-3beta activity associated with a marked reduction of beta-cell proliferation and increased apoptosis. Irs2-/- mice crossed with Gsk-3beta+/- mice preserved beta-cell mass by reversing the negative effects on proliferation and apoptosis, preventing onset of diabetes. Previous studies had shown that islets of Irs2-/- mice had increased cyclin-dependent kinase inhibitor p27(kip1) that was limiting for beta-cell replication, and reduced Pdx1 levels associated with increased cell death. Preservation of beta-cell mass in Gsk-3beta+/- Irs2-/- mice was accompanied by suppressed p27(kip1) levels and increased Pdx1 levels. To separate peripheral versus beta-cell-specific effects of reduction of Gsk3beta activity on preservation of beta-cell mass, mice homozygous for a floxed Gsk-3beta allele (Gsk-3(F/F)) were then crossed with rat insulin promoter-Cre (RIP-Cre) mice to produce beta-cell-specific knockout of Gsk-3beta (betaGsk-3beta-/-). Like Gsk-3beta+/- mice, betaGsk-3beta-/- mice also prevented the diabetes of the Irs2-/- mice. The results of these studies now define a new, negatively regulated substrate of the insulin signaling pathway specifically within beta-cells that when elevated, can impair replication and increase apoptosis, resulting in loss of beta-cells and diabetes. These results thus form the rationale for developing agents to inhibit this enzyme in obese insulin-resistant individuals to preserve beta-cells and prevent diabetes onset.


Multicenter Advanced Pediatric Otolaryngology Fellowship Prep Surgical Simulation Course with 3D Printed High-Fidelity Models.
To assess the effect of 3-dimensional (3D)-printed surgical simulators used in an advanced pediatric otolaryngology fellowship preparatory course on trainee education. Quasi-experimental pre/postsurvey. Multicenter collaborative course conducted at a contract research organization prior to a national conference. A 5-station, 7-simulator prep course was piloted for 9 pediatric otolaryngology fellows and 17 otolaryngology senior residents, with simulators for airway graft carving, microtia ear framework carving, and cleft lip/palate repair. Prior to the course, trainees were provided educational materials electronically along with presurveys rating confidence, expertise, and attitude around surgical simulators. In October 2018, surgeons engaged in simulation stations with direction from 2 attending faculty per station, then completed postsurveys for each simulator. Statistically significant increases (P < .05) in self-reported confidence (average, 53%; range, 18%-80%) and expertise (average, 68%; range, 9%-95%) were seen across all simulators, corresponding to medium to large effect sizes as measured by Cohen's d statistic (0.41-1.71). Positive attitudes around 3D printing in surgical education also demonstrated statistically significant increases (average, 10%; range, 8%-13%). Trainees commented positively on gaining such broad exposure, although consistently indicated a preference for more practice time during the course. We demonstrate the benefit of high-fidelity, 3D-printed simulators in exposing trainees to advanced procedures, allowing them hands-on practice in a zero-risk environment. In the future, we hope to refine this course design, develop standardized tools to assess their educational value, and explore opportunities for integration into use in milestone assessment and accreditation.


Testing the phenotypic gambit: phenotypic, genetic and environmental correlations of colour.
Evolutionary theory is primarily concerned with genetic processes, yet empirical testing of this theory often involves data collected on phenotypes. To make this tenable, the implicit assumption is often made that phenotypic patterns are good predictors of genetic patterns; an assumption that coined the phenotypic gambit. Although this assumption has been validated for traits with high heritability, such as morphology, its generality for traits with low heritabilities, such as life-history and behavioural traits, remains controversial. Using a large-scale cross-fostering experiment, we were able to measure genetic, common environmental and phenotypic correlations between four colour traits and two skeletal traits in a wild population of passerine birds, the blue tit (Parus caeruleus). Colour traits had little heritable variation but common environment effects were found to be important; skeletal traits showed the opposite pattern. Positive correlations because of a shared natal environment were found between all traits, obscuring negative genetic correlations between some colour and skeletal traits. Consequently, phenotypic patterns were poor surrogates for genetic patterns and we suggest that this may be common if trade-offs or substantial parental effects exist. For this group of traits, the phenotypic gambit cannot be made and we suggest caution when inferring genetic patterns from phenotypic data, especially for behavioural and life-history traits.


Distinct biogenesis pathways may have led to functional divergence of the human and Drosophila Arglu1 sisRNA.
Stable intronic sequence RNAs (sisRNAs) are stable, long noncoding RNAs containing intronic sequences. While sisRNAs have been found across diverse species, their level of conservation remains poorly understood. Here we report that the biogenesis and functions of a sisRNA transcribed from the highly conserved Arglu1 locus are distinct in human and Drosophila melanogaster. The Arglu1 genes in both species show similar exon-intron structures where the intron 2 is orthologous and positionally conserved. In humans, Arglu1 sisRNA retains the entire intron 2 and promotes host gene splicing. Mechanistically, Arglu1 sisRNA represses the splicing-inhibitory activity of ARGLU1 protein by binding to ARGLU1 protein and promoting its localization to nuclear speckles, away from the Arglu1 gene locus. In contrast, Drosophila dArglu1 sisRNA forms via premature cleavage of intron 2 and represses host gene splicing. This repression occurs through a local accumulation of dARGLU1 protein and inhibition of telescripting by U1 snRNPs at the dArglu1 locus. We propose that distinct biogenesis of positionally conserved Arglu1 sisRNAs in both species may have led to functional divergence.


Human lipodystrophies: genetic and acquired diseases of adipose tissue.
Human lipodystrophies represent a heterogeneous group of diseases characterized by generalized or partial fat loss, with fat hypertrophy in other depots when partial. Insulin resistance, dyslipidemia and diabetes are generally associated, leading to early complications. Genetic forms are uncommon: recessive generalized congenital lipodystrophies result in most cases from mutations in the genes encoding seipin or the 1-acyl-glycerol-3-phosphate-acyltransferase 2(AGPAT2). Dominant partial familial lipodystrophies result from mutations in genes encoding the nuclear protein lamin A/C or the adipose transcription factor PPARgamma. Importantly, lamin A/Cmutations are also responsible for metabolic laminopathies, resembling the metabolic syndrome and progeria, a syndrome of premature aging. A number of lipodystrophic patients remain undiagnosed at the genetic level. Acquired lipodystrophy can be generalized, resembling congenital forms, or partial, as the Barraquer-Simons syndrome, with loss of fat in the upper part of the body contrasting with accumulation in the lower part. Although their etiology is generally unknown, they could be associated with signs of autoimmunity. The most common forms of lipodystrophies are iatrogenic. In human immunodeficiency virus-infected patients, some first-generation antiretroviral drugs were strongly related with peripheral lipoatrophy and metabolic alterations. Partial lipodystrophy also characterize patients with endogenous or exogenous long-term corticoid excess. Treatment of fat redistribution can sometimes benefit from plastic surgery. Lipid and glucose alterations are difficult to control leading to early occurrence of diabetic, cardiovascular and hepatic complications.


5-[123I/125I]iodo-2'-deoxyuridine in metastatic lung cancer: radiopharmaceutical formulation affects targeting.
This study assesses targeting of lung metastases in mice with the radioiodinated thymidine analog 5-[(123)I/(125)I]iodo-2'-deoxyuridine ((123)I-IUdR/(125)I-IUdR), formulated with varying amounts of tributyltin precursor and injected intravenously. Six- to 8-wk-old C57BL/6 mice were injected intravenously with B16F10 melanoma cells. Two weeks later, when lung tumors were established, the animals were injected intravenously with (125)I-IUdR synthesized using 1, 35, 100, 150, 200, or 250 microg 5-tributylstannyl-2'-deoxyuridine (SnUdR) in the presence of an oxidant. Nontumor-bearing mice were also injected with these formulations and served as control animals. Twenty-four hours later, the animals were killed, and the radioactivity associated with the lungs and other tissues was measured in a gamma-counter. The percentage injected dose per gram tissue (%ID/g) and tumor-to-nontumor ratios (T/NT ratios) were calculated. Phosphor imaging was done on lungs from tumor-bearing and nontumor-bearing mice injected with (125)I-IUdR formulated with each tin precursor concentration. Scintigraphy was also performed 3 and 24 h after intravenous injection of (123)I-IUdR. The %ID/g (125)I-IUdR was higher in lungs of tumor-bearing animals than in lungs of control animals. Although the increase in SnUdR present led to a small but statistically significant decrease in the radioactive content of normal lungs, a 3-fold increase was observed in the lungs of tumor-bearing animals with radiopharmaceutical formulated with 100 microg SnUdR (5 microg per mouse). This enhancement in radioactive uptake by the lungs led to approximately 14-fold increases in T/NT ratios. Phosphor imaging ((125)I-IUdR) of lungs as well as scintigraphy ((123)I-IUdR) of whole animals substantiated these findings. The formulation for the synthesis of radio-IUdR that leads to the highest %ID/g in tumor and the best T/NT ratio has been identified. Further studies are required to determine the factors responsible for specific enhancement in IUdR tumor uptake.


Percutaneous aortic valve replacement: emerging tractability for sufficient intracardiac resection of the aortic valve.
: The feasibility of endovascular resection of highly calcified aortic valves has already been demonstrated by our group. Different endovascular and intracardiac tractability methods were applied. In this study, these technologies were analyzed comparing the tractability, the resection time, and the lesions in the surrounding tissue. : All aortic valve resections (seven human hearts and 21 porcine hearts) were performed using a Thulium:YAG laser (continuous wave, wavelength of 2.01 μm, 20 watts power rating). In the first resection system, the laser fiber was controlled by a free in-lying flexible endoscope (Ø 2.5 mm, length of 600 mm). The distal part of the endoscope (40 mm) was moved in one plane by proximal manual control (three degrees of freedom). The resection system was separated into defined rooms assigning one room for one tool. The fiber was controlled by the above-mentioned endoscope (*) (three degrees of freedom). The third resection system was a mechanical microactuator carrying the laser fiber (three degrees of freedom). The fourth resection system contains a rotatable inlay with defined rooms and a newly designed nitinol (NiTi) microactuator that controlled the laser fiber (four degrees of freedom). The resection time per leaflet was measured in minutes. Gross anatomy and histology in the surrounding tissue were evaluated. : The resection time in approaches 1, 2, 3, and 4 was 5.5 ± 2.3 minutes, 7.4 ± 2.7 minutes, ± 6.6 minutes, and2.3 ± 1.2 minutes, respectively. The gross anatomy and histology of collateral damages revealed only superficial lesions of the surrounding tissue. The amount of lesions and the resection time were lower in the fourth approach with four degrees of freedom. : This analysis demonstrated that a precise tractability with four degrees of freedom is necessary for a faster and safer endovascular resection of the aortic valve. The analysis will help to optimize the ongoing development of the endovascular and intracardiac resection technology.


Two decades of external peer review of cancer care in general hospitals; the Dutch experience.
External peer review was introduced in general hospitals in the Netherlands in 1994 to assess and improve the multidisciplinary team approach in cancer care. This paper aims to explore the value, perceived impact, and (future) role of external peer review in cancer care. Semistructured interviews were held with clinicians, oncology nurses, and managers from fifteen general hospitals that participated in three rounds of peer review over a period of 16 years. Interviewees reflected on the goals and expectations, experiences, perceived impact, and future role of external peer review. Transcriptions of the interviews were coded to discover recurrent themes. Improving clinical care and organization were the main motives for participation. Positive impact was perceived on multiple aspects of care such as shared responsibilities, internal prioritization of cancer care, improved communication, and a clear structure and position of cancer care within general hospitals. Establishing a direct relationship between the external peer review and organizational or clinical impact proved to be difficult. Criticism was raised on the content of the program being too theoretical and organization-focussed after three rounds. According to most stakeholders, external peer review can improve multidisciplinary team work in cancer care; however, the acceptance is threatened by a perceived disbalance between effort and visible clinical impact. Leaner and more clinically focused programs are needed to keep repeated peer reviews challenging and worthwhile.


Comparison of film dosimetry techniques used for quality assurance of intensity modulated radiation therapy.
Accurate dosimetry is essential to ensure the quality of advanced radiation treatments, such as intensity modulated radiation therapy (IMRT). Therefore, a comparison study was conducted to assess the accuracy of various film dosimetry techniques that are widely used in clinics. A simulated IMRT plan that produced an inverse pyramid dose distribution in a perpendicular plane of the beam axis was designed with 6 MV x rays to characterize the large contribution of scattered photons to low dose regions. Three film dosimetry techniques, EDR2, EDR2 with low-energy photon absorption lead filters (EDR2 WF), and GafChromic EBT, were compared to ionization chamber measurements as well as Monte Carlo (MC) simulations. The accuracy of these techniques was evaluated against the ionization chamber data. Two-dimensional comparisons with MC simulation results were made by computing the gamma index (gamma) with criteria ranging from 2% of dose difference or 2 mm of distance to agreement (2%/2 mm) to 4%/4 mm on the central vertical plane (20 x 20 cm2) of a square solid water phantom. Depth doses and lateral profiles at depths of 5, 10, and 15 cm were examined to characterize the deviation of film measurements and MC predictions from ionization chamber measurements. In depth dose comparisons, the deviation between the EDR2 films was 9% in the low dose region and 5% in high dose region, on average. With lead filters, the average deviation was reduced to -1.3% and -0.3% in the low dose and high dose regions, respectively. EBT film results agreed within 1.5% difference on average with ionization chamber measurements in low and high dose regions. In two-dimensional comparisons with MC simulation, EDR2 films passed gamma tests with a 2%/2 mm criterion only in the high dose region (gamma < or = 1, total of 63.06% of the tested region). In the low dose region, EDR2 films passed gamma tests with 3%/3 mm criterion (gamma < or = 1, total of 98.4% of the tested region). For EDR2 WF and GafChromic EBT films, gamma tests with a 2% /2 mm criterion (gamma < or = 1) in the tested area was 97.3% and 96.8% of the tested region, respectively. The EDR2 film WF and GafChromic EBT film achieved an average accuracy level of 1.5% against an ionization chamber. These two techniques agreed with the MC prediction in 2%/2mm criteria evaluated by the gamma index, whereas EDR2 without filters achieved an accuracy level of 3%/3 mm with the decision criteria of agreement greater than 95% of the tested region. The overall results will provide a useful quantitative reference for IMRT verifications.


A Molecular Investigation of Malaria Infections From High-Transmission Areas of Southern Togo Reveals Different Species of Plasmodium Parasites.
Malaria particularly burdens people in poor and neglected settings across the tropics of Africa. Meanwhile, a large proportion of the Togo population have poor understanding of malaria epidemiology and parasites. This study carried out a molecular survey of malaria cases in southern Togo during 2017-2019. We estimated Plasmodium species infection rates and microscopic examination compliance with nested PCR results. Sensitivity and specificity analyses were performed in conjunction with predictive values. Also, phylogenetic characterization of species of malaria parasites was assessed. Plasmodium genus-specific nested PCR identified 565 positive cases including 536/611 (87.8%) confirmed cases from the microscopy-positive group and 29/199 (14.6%) diagnosed malaria cases from the microscopy-negative group. Our findings revealed a disease prevalence (69.8%) higher than that reported (25.5-55.1%) for the country. The diagnostic test had 94.9% sensitivity and 69.4% specificity, i.e., it missed 120 of the people who had malaria and about one-third of the people tested positive for the disease, which they did not have, respectively. In conjunction, the test showed 87.7% positive predictive value and 85.4% negative predictive value, which, from a clinical perspective, indicates the chance that a person with a positive diagnostic test truly has the disease and the probability that a person with a negative test does not have the disease, respectively. Further species-specific nested PCR followed by analysis of gene sequences confirmed species of malaria parasites and indicated infection rates for Plasmodium falciparum (Pf), 95.5% (540/565); P. ovale (Po), 0.5% (3/565); and P. malariae (Pm), 0.4% (2/565). In addition, 20 cases were coinfection cases of Pf-Po (15/565) and Pf-Pm (5/565). This study publicly reports, for the first time, a molecular survey of malaria cases in Togo and reveals the presence of other malaria parasites (Po and Pm) other than Pf. These findings might provide answers to some basic questions on the malaria scenario and, knowledge gained could help with intervention deployment for effective malaria control in Togo.


Oxidative Stress Induction Is a Rational Strategy to Enhance the Productivity of Antrodia cinnamomea Fermentations for the Antioxidant Secondary Metabolite Antrodin C.
Antioxidant metabolites contribute to alleviating oxidative stress caused by reactive oxygen species (ROS) in microorganisms. We utilized oxidative stressors such as hydrogen peroxide supplementation to increase the yield of the bioactive secondary metabolite antioxidant antrodin C in submerged fermentations of the medicinal mushroom Antrodia cinnamomea. Changes in the superoxide dismutase and catalase activities of the cells indicate that ROS are critical to promote antrodin C biosynthesis, while the ROS production inhibitor diphenyleneiodonium cancels the productivity-enhancing effects of H2O2. Transcriptomic analysis suggests that key enzymes in the mitochondrial electron transport chain are repressed during oxidative stress, leading to ROS accumulation and triggering the biosynthesis of antioxidants such as antrodin C. Accordingly, rotenone, an inhibitor of the electron transport chain complex I, mimics the antrodin C productivity-enhancing effects of H2O2. Delineating the steps connecting oxidative stress with increased antrodin C biosynthesis will facilitate the fine-tuning of strategies for rational fermentation process improvement.


Compositional and abundance changes of nitrogen-cycling genes in plant-root microbiomes along a salt marsh chronosequence.
Disentangling the relative influences of soil properties and plant-host on root-associated microbiomes in natural systems is challenging, given that spatially segregated soil types display distinct historical legacies. In addition, distant locations may also lead to biogeographical patterns of microbial communities. Here, we used an undisturbed salt marsh chronosequence spanning over a century of ecosystem development to investigate changes in the community composition and abundance of a set of nitrogen-cycling genes. Specifically, we targeted genes of diazotrophs and ammonia oxidizers associated with the bulk and rhizosphere soil of the plant species Limonium vulgare. Samples were collected across five distinct successional stages of the chronosequence (ranging from 5 to 105 years) at two time-points. Our results indicate that soil variables such as sand:silt:clay % content and pH strongly relates to the abundance of N-cycling genes in the bulk soil. However, in the rhizosphere samples, the abundance of ammonia-oxidizing organisms (both bacteria and archaea, AOB and AOA, respectively) was relatively constant across most of the successional stages, albeit displaying seasonal variation. This result indicates a potentially stronger control of plant host (rather than soil) on the abundance of these organisms. Interestingly, the plant host did not have a significant effect on the composition of AOA and AOB communities, being mostly divergent according to soil successional stages. The abundance of diazotrophic communities in rhizosphere samples was more affected by seasonality than those of bulk soil. Moreover, the abundance pattern of diazotrophs in the rhizosphere related to the systematic increase of plant biomass and soil organic matter along the successional gradient. These results suggest a potential season-dependent regulation of diazotrophs exerted by the plant host. Overall, this study contributes to a better understanding of how the natural formation of a soil and host plants influence the compositional and abundance changes of nitrogen-cycling genes in bulk and rhizosphere soil microhabitats.


Tracing organizing principles: learning from the history of systems biology.
With the emergence of systems biology, the identification of organizing principles is being highlighted as a key research aim. Researchers attempt to "reverse engineer" the functional organization of biological systems using methodologies from mathematics, engineering and computer science while taking advantage of data produced by new experimental techniques. While systems biology is a relatively new approach, the quest for general principles of biological organization dates back to systems theoretic approaches in early and mid-twentieth century. The aim of this paper is to draw on this historical background in order to increase the understanding of the motivation behind the search for general principles and to clarify different epistemic aims within systems biology. We pinpoint key aspects of earlier approaches that also underlie the current practice. These are i) the focus on relational and system-level properties, ii) the inherent critique of reductionism and fragmentation of knowledge resulting from overspecialization, and iii) the insight that the ideal of formulating abstract organizing principles is complementary to, rather than conflicting with, the aim of formulating detailed explanations of biological mechanisms. We argue that looking back not only helps us understand the current practice but also points to possible future directions for systems biology.


Fundamental Frequency in Excised Human Larynges after Anterior Glottic Web Formation of Various Extents.
To determine the effect of vocal fold anterior web formation on fundamental frequency with a cadaveric excised larynx model. Experimental study with excised human larynges. Academic tertiary care hospital. Sixteen freshly excised human larynges were evaluated with high-speed videoendoscopy and digital kymography during artificially produced vibration. Each larynx was assessed in 4 conditions: preoperative controls and after 25%, 33%, and 50% decreases in the vibratory portion of the vocal folds. The following parameters were evaluated: fundamental frequency, periodicity, vocal fold vibration amplitude, phase symmetry, and glottic closure. The mean fundamental frequencies were 208.87, 250.20, 292.37, and 342.67 Hz for preoperative controls and 25%, 33%, and 50% reductions in vibratory length of the vocal folds, respectively. Fundamental frequency increased with each increase in anterior glottic web extent, and the difference among the groups was statistically significant in absolute values in hertz and in semitone elevation. The mathematical models for estimating postoperative fundamental frequency had a statistically significant coefficient. The vibration of the vocal folds remained periodic in all larynges before and after the procedures. There was a significant and progressive increase in the fundamental frequency with each enlargement of anterior glottic web. Based on the control frequency, mathematical models could estimate the value of the fundamental frequency after the procedure.


Partial mixture model for tight clustering of gene expression time-course.
Tight clustering arose recently from a desire to obtain tighter and potentially more informative clusters in gene expression studies. Scattered genes with relatively loose correlations should be excluded from the clusters. However, in the literature there is little work dedicated to this area of research. On the other hand, there has been extensive use of maximum likelihood techniques for model parameter estimation. By contrast, the minimum distance estimator has been largely ignored. In this paper we show the inherent robustness of the minimum distance estimator that makes it a powerful tool for parameter estimation in model-based time-course clustering. To apply minimum distance estimation, a partial mixture model that can naturally incorporate replicate information and allow scattered genes is formulated. We provide experimental results of simulated data fitting, where the minimum distance estimator demonstrates superior performance to the maximum likelihood estimator. Both biological and statistical validations are conducted on a simulated dataset and two real gene expression datasets. Our proposed partial regression clustering algorithm scores top in Gene Ontology driven evaluation, in comparison with four other popular clustering algorithms. For the first time partial mixture model is successfully extended to time-course data analysis. The robustness of our partial regression clustering algorithm proves the suitability of the combination of both partial mixture model and minimum distance estimator in this field. We show that tight clustering not only is capable to generate more profound understanding of the dataset under study well in accordance to established biological knowledge, but also presents interesting new hypotheses during interpretation of clustering results. In particular, we provide biological evidences that scattered genes can be relevant and are interesting subjects for study, in contrast to prevailing opinion.


The histology of subcutaneously implanted donor bronchial rings correlates with rejection scores of lung allografts in a primate lung transplant model.
The diagnosis of acute rejection in lung transplantation generally relies on transbronchial biopsies. This invasive procedure may be associated with bronchial bleeding or pneumothorax and may not be feasible in patients with severely compromised lung function. The hypothesis of the current study was that histopathological findings of donor bronchial segments implanted into the subcutaneous tissue of lung allograft recipients would predict lung tissue rejection scores, thus providing the clinician with an alternate source of information. Unilateral left lung transplantation was performed in 34 cynomolgus monkeys as part of a drug efficacy study. After completion of the transplant procedure, 4 bronchial ring segments of the explanted recipient left lung and 4 bronchial ring segments of the non-transplanted right donor lung were implanted subcutaneously in the abdominal region. Lung allograft rejection was evaluated by open lung biopsies of the allograft performed on postoperative (PO) Day 14 and during sacrifice on PO Day 28. At the time of each biopsy, 2 donor and 2 recipient subcutaneous bronchial rings were explanted. Histologic evaluation of the lung tissue samples was performed according to the working formulation of the International Society for Heart and Lung Transplantation. Bronchial rings were independently evaluated by assessing the degree of airway narrowing; percentage of intact epithelial coverage as well as its specific histology (respiratory ciliated, flattened cuboidal, squamous); presence of lymphocytes, macrophages or spindle cells; and presence of peribronchial inflammation, luminal fibrosis, lymphocytic bronchitis or luminal mucous. Statistical analysis was performed by logistic regression. In the recipient bronchial rings, there was no evidence of airway narrowing. There was 98% epithelial coverage, 71% that were respiratory ciliated cells, and there was no inflammation. Donor bronchial rings showed no airway narrowing for monkeys with grade A0 to A2 rejection in tissue biopsies and a maximum narrowing (41.2%) with A4 rejection. Epithelial cell coverage was approximately 100% with grade A0-A2 and 44+/-11% with A4 rejection. Lymphocytic bronchitis was most severe in A4 rejection and minimal in A0 to A2 rejection. By logistic regression analysis, independent predictors of a likelihood of rejection were the degree of airway obliteration, the percentage of epithelial cell coverage, the degree of lymphocytic bronchitis and the product of respiratory and flattened cuboidal cell coverage. The current data show that histologic alterations of subcutaneously implanted donor bronchial rings correlate with lung tissue biopsy scores based on the ISHLT working formulation. Because subcutaneous bronchial rings can be explanted under local anesthesia, they may provide useful information for the diagnosis of acute allograft rejection in patients with impaired lung function, patients that obtaining lung tissue samples may not be feasible.


Hybrid approach for coronary artery revascularization: where do we stand?
Recently, hybrid coronary artery revascularization (HCAR), combining the benefits of both percutaneous coronary intervention and coronary artery bypass graft surgery (CABG) while minimizing their respective shortcomings, has been developed. This review is aimed to explore and discuss recent clinical outcomes and patient selection, and comment on surgical approaches for HCAR. Current forms of HCAR include off-pump mini-sternotomy or on-pump full sternotomy CABG [left internal mammary artery (LIMA)-to-left anterior descending artery(LAD) CABG followed by drug-eluting stents (DES) to non-LAD territories], robotic-assisted off-pump HCAR (robotic LIMA-to-LAD CABG and DES to non-LAD territories), and off-pump mini-thoracotomy single-vessel small thoracotomy (LIMA-to-LAD CABG), all of which have reported acceptable early to mid-term patency rates and freedom from major cardiac and cerebrovascular adverse events. As long-term effectiveness compared with conventional CABG remains to be demonstrated, especially in patients with diabetes and patients with higher SYNTAX scores, appropriate discussion between the 'Heart Team' and patient is needed prior to HCAR. HCAR presents an attractive alternative option for treating patients with multivessel coronary artery disease because it maximizes the clear survival benefits of LIMA-LAD grafting, improves quality assurance with completion angiography, and allows quicker patient recovery; furthermore, patients avoid the negative systemic inflammatory effects of cardiopulmonary bypass and delayed healing after sternotomy.


UBAP2L is amplified in a large subset of human lung adenocarcinoma and is critical for epithelial lung cell identity and tumor metastasis.
The ubiquitin-associated protein 2-like (UBAP2L) gene remains poorly studied in human and mouse development. UBAP2L interacts with the Polycomb group protein B lymphoma Mo-MLV insertion region 1 homolog (BMI1) and determines the activity of mouse hematopoietic stem cells in vivo Here we show that loss of Ubap2l leads to disorganized respiratory epithelium of mutant neonates, which die of respiratory failure. We also show that UBAP2L overexpression leads to epithelial-mesenchymal transition-like phenotype in a non-small cell lung carcinoma (NSCLC) cell line. UBAP2L is amplified in 15% of human primary lung adenocarcinoma specimens. Such patients express higher levels of UBAP2L and show a reduction in survival when compared with those who do not have this gene amplification. Supporting a possible role for UBAP2L in lung tumor progression, NSCLC cells engineered to express low levels of this gene produce much smaller tumors in vivo than wild-type control cells. Together, these results suggest that UBAP2L contributes to epithelial lung cell identity in mice and that it plays an important role in human lung adenocarcinoma.-Aucagne, R., Girard, S., Mayotte, N., Lehnertz, B., Lopes-Paciencia, S., Gendron, P., Boucher, G., Chagraoui, J., Sauvageau, G. UBAP2L is amplified in a large subset of human lung adenocarcinoma and is critical for epithelial lung cell identity and tumor metastasis.


Serum markers of type III and IV procollagen processing predict recurrence of fibrosis in liver transplanted patients.
Following liver transplantation (LT), 10-30% of patients develop recurrent cirrhosis (RC). There is an urgent need for predictive non-invasive markers for improved monitoring of these patients. Here we studied extracellular matrix biomarkers as predictors of RC after LT. Forty-seven LT patients were divided into groups of fast, intermediate or non-progressors towards RC (<1 year, 3-5 years or no advanced fibrosis >5 years after LT), assessed by follow-up liver biopsies. Markers of interstitial matrix type III and V collagen formation (PRO-C3 and PRO-C5), basement membrane type IV collagen formation (PRO-C4) and degradation (C4M) were assessed in serum samples collected 3, 6 and 12 months post-LT using specific ELISAs. PRO-C3, PRO-C4, and C4M were elevated in fast progressors compared to non-progressors 3 months after LT. C4M and PRO-C4 additionally differentiated between intermediate and fast progressors at 3 months. PRO-C3 was best predictor of survival, with LT patients in the highest PRO-C3 tertile having significantly shorter survival time. This shows that interstitial matrix and basement membrane remodeling in RC may be distinguishable. Markers originating from different sites in the extracellular matrix could be valuable tools for a more dynamic monitoring of patients at risk of RC. However, this needs validation in larger cohorts.


An evidence-based approach to understanding the competency development needs of the health service management workforce in Australia.
Competent managers are essential to the productivity of organisations and the sustainability of health systems. Effective workforce development strategies sensitive to the current competency development needs of health service managers (HSMs) are required. To conduct a 360° assessment of the competence of Australian HSMs to identify managerial competence levels, and training and development needs. Assessment of 93 middle-level HSMs from two public hospitals (n = 25) and five community health services (CHS) (n = 68), using the Managerial Competency Assessment Partnership (MCAP) framework and tool, conducted between 2012 and 2014 in Victoria, Australia. Mean competency scores from both self- and combined colleagues' assessments indicated competence (scores greater than five but less than six) without guidance, but many HSMs have not had extensive experience. Around 12% of HSMs were unable to demonstrate the competency of 'evidence-informed decision-making' and 4% of HSMs were unable to demonstrate the competency of 'enabling and managing change'. The assessments confirmed managerial competence for the majority of middle-level HSMs from hospitals and CHS in Victoria, but found competency gaps. In addition, the assessment confirmed managerial strengths and weaknesses varied across management groups from different organisations. These findings suggest that the development of strategies to strengthen the health service management workforce should be multifaceted. A focus on competency in performance evaluation and development using the MCAP framework and tool not only provides insights into performance of HSMs, but also has the potential to provide an organisation strategic advantage through succession planning and advancing managers' competence via learning needs analysis and targeted professional development. Linking competencies of HSMs to organisational objectives and strategies provides optimal use of the human resource capacity, improving the organisation's productivity and sustainability.


Pharmacology of a Potent and Novel Inhibitor of the NOD-Like Receptor Pyrin Domain-Containing Protein 3 (NLRP3) Inflammasome that Attenuates Development of Nonalcoholic Steatohepatitis and Liver Fibrosis.
The NOD-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome is a multiprotein complex and component of the innate immune system that is activated by exogenous and endogenous danger signals to promote activation of caspase-1 and the maturation and release of the proinflammatory cytokines interleukin (IL)-1β and IL-18. Inappropriate activation of NLRP3 has been implicated in the pathophysiology of multiple inflammatory and autoimmune diseases, including cardiovascular disease, neurodegenerative diseases, and nonalcoholic steatohepatitis (NASH), thus increasing the clinical interest of this target. We describe in this study the preclinical pharmacologic, pharmacokinetic, and pharmacodynamic properties of a novel and highly specific NLRP3 inhibitor, JT001 (6,7-dihydro-5H-pyrazolo[5,1-b][1,3]oxazine-3-sulfonylurea). In cell-based assays, JT001 potently and selectively inhibited NLRP3 inflammasome assembly, resulting in the inhibition of cytokine release and the prevention of pyroptosis, a form of inflammatory cell death triggered by active caspase-1. Oral administration of JT001 to mice inhibited IL-1β production in peritoneal lavage fluid at plasma concentrations that correlated with mouse in vitro whole blood potency. Orally administered JT001 was effective in reducing hepatic inflammation in three different murine models, including the Nlrp3A350V /+CreT model of Muckle-Wells syndrome (MWS), a diet-induced obesity NASH model, and a choline-deficient diet-induced NASH model. Significant reductions in hepatic fibrosis and cell damage were also observed in the MWS and choline-deficient models. Our findings demonstrate that blockade of NLRP3 attenuates hepatic inflammation and fibrosis and support the use of JT001 to investigate the role of NLRP3 in other inflammatory disease models. SIGNIFICANCE STATEMENT: Persistent inflammasome activation is the consequence of inherited mutations of NLRP3 and results in the development of cryopyrin-associated periodic syndromes associated with severe systemic inflammation. NLRP3 is also upregulated in nonalcoholic steatohepatitis, a metabolic chronic liver disease currently missing a cure. Selective and potent inhibitors of NLRP3 hold great promise and have the potential to overcome an urgent unmet need.


Time course of and stimuli to compensatory growth of the lung after pneumonectomy.
Left pneumonectomy in the mature rat led to an increase of [3Ha1 thymidine incorporation into DNA of the remaining lung in the first 3 postoperative days, and resulted in a subsequent 38% increase of lung weight and 41% increase of lung tissue volume measured 1 wk after surgery. Despite these early changes, total lung volume (TLV) did not increase until the 2nd postoperative wk, reaching values 33% greater than in controls. Analysis of lung pressure-volume curves revealed that lung recoil was increased at low lung volumes 1 wk after surgery, but returned to normal by the 2nd postoperative wk, suggesting that synthesis of both lung elastin and collagen had occurred by this time. Increased inspired oxygen concentration (28% or 35%) during the 1st but not the 2nd postoperative wk abolished the change in TLV without influencing the increase in lung weight, while diminished inspired oxygen (17% or 14%) accentuated the postoperative increase in TLV. Lung pressure-volume curves demonstrated changes in distensibility at low lung volumes, suggesting that oxygen may have influenced synthesis or cross-linking of lung elastin. Alterations of minute ventilation in the postoperative period produced by 3% CO2 did not influence the compensatory growth process, nor did administration of cyclophosphamide. These studies suggest that postpneumonectomy lung growth is a two-phase process, beginning with cell proliferation and increased tissue volume, followed by increasing lung volume associated with formation of lung structural proteins. The latter process is profoundly influenced by inspired oxygen concentration in the early postoperative period.


Anamorphic transformation and its application to time-bandwidth compression.
A general method for compressing the modulation time-bandwidth product of analog signals is introduced. As one of its applications, this physics-based signal grooming, performed in the analog domain, allows a conventional digitizer to sample and digitize the analog signal with variable resolution. The net result is that frequency components that were beyond the digitizer bandwidth can now be captured and, at the same time, the total digital data size is reduced. This compression is lossless and is achieved through a feature selective reshaping of the signal's complex field, performed in the analog domain prior to sampling. Our method is inspired by operation of Fovea centralis in the human eye and by anamorphic transformation in visual arts. The proposed transform can also be performed in the digital domain as a data compression algorithm to alleviate the storage and transmission bottlenecks associated with "big data."


Diethylstilbestrol effects on serum enzymes and isozymes in muscular dystrophy.
Diethyistibestrol was administered orally to 11 boys with Duchene muscular dystrophy (DMD). Creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) activities, characteristically high in DMD, and presumably of muscle origin, were reduced significantly (P LESS THAN .05). On ther other hand, the activity of alkaline phosphatase, an enzyme not of muscle origin, increased. These enzyme changes were reversible when diethyistibestrol was discontinued. Despite appreciable alterations in totoal serum enzyme activity, no important change was found in the isozyme patterns. Piperazine estrone sulfate was administered to another patient with DMD. The effects of this physiologic hormone were, in part, similar to those of diethyistibestrol. Experimentally, diethylstilbestrol reduced the efflux of CPK and LDH from mouse skeletal muscle. This may be the manner by which diethylstilbesterol reduced the serum enzyme levels in DMD, but this has not been proved directly. These studies are the first step in an effort to identify various agents that in combinations may normalize serum enzyme levels.


Identification of candidate microRNA markers of endometriosis with the use of next-generation sequencing and quantitative real-time polymerase chain reaction.
To identify novel candidate diagnostic microRNA (miRNA) markers of endometriosis by means of an unbiased search with confirmation by means of targeted polymerase chain reaction (PCR). Retrospective cohort. University teaching hospitals. Women with endometriosis and control women, confirmed with the use of laparoscopy. Diagnostic laparoscopy and blood sample. Next-generation sequencing (NGS) and quantitative real-time PCR (qRT-PCR). Candidate miRNAs differentially expressed in women with endometriosis compared with control women were identified by means of NGS and selected for qRT-PCR. Plasma samples from another cohort of women with surgically confirmed endometriosis (n = 53) and disease-free control women (n = 53) were checked for hemolysis using spectrophotometry and the ratio of miR-23a and miR-451 by means of qRT-PCR. MicroRNA signatures were quantified by means of qRT-PCR in hemolysis-free plasma samples of case subjects (n = 25) and control subjects (n = 28) with the use of miRcury LNA miRNA. Circulating levels of eight miRNAs (miR-199a-3p, miR-143-3p, miR-340-5p, let-7b-5p, miR-21-5p, miR-17-5p, miR-20a-5p, and miR-103a-3p) were significantly lower in case subjects compared to control subjects. The sensitivity and specificity for individual miRNAs ranged from 0.36 to 1.00 and from 0.43 to 1.00, respectively, but when combined produced sensitivity and specificity of 0.92 and 0.86 with positive (PPV) and (NPV) predictive values of 0.85 and 0.92, respectively. However, combination of five miRNAs (miR-17-5p, miR-20a-5p, miR-199a-3p, miR-143-3p, and let-7b-5p) produced sensitivity and specificity of 0.96 and 0.79 with PPV and NPV of 0.80 and 0.96, respectively. We conclude that a panel of candidate miRNAs was comparable to laparoscopy in distinguishing between women with endometriosis and control women.


Pathological findings correlated with magnetic resonance imaging in subcortical arteriosclerotic encephalopathy (Binswanger's disease).
Formalin-fixed brain slices from four cases of subcortical arteriosclerotic encephalopathy in which a firm diagnosis could be made both clinically and pathologically have been studied by magnetic resonance imaging (MRI). The slices were subsequently embedded in paraffin-wax or celloidin and sections were cut in the same plane as the MRI slices. There was a good correlation between the extent and severity of the abnormal MRI signal and the pathological changes. Areas of diffuse MRI abnormality corresponded with areas of axonal and myelin loss with gliosis, and small "lacune"-like lesions corresponded with lacunar infarcts histologically. Sparing of the subcortical U-fibres was seen histologically and on MRI. The abnormal signal probably originates from increased tissue water attributable to gliosis and an expanded extracellular space.


Protein binder for affinity purification of human immunoglobulin antibodies.
The importance of a downstream process for the purification of immunoglobulin antibodies is increasing with the growing application of monoclonal antibodies in many different areas. Although protein A is most commonly used for the affinity purification of antibodies, certain properties could be further improved: higher stability in alkaline solution and milder elution condition. Herein, we present the development of Fc-specific repebody by modular engineering approach and its potential as an affinity ligand for purification of human immunoglobulin antibodies. We previously developed the repebody scaffold composed of Leucine-rich repeat (LRR) modules. The scaffold was shown to be highly stable over a wide range of pH and temperature, exhibiting a modular architecture. We first selected a repebody that binds the Fc fragment of human immunoglobulin G (IgG) through a phage display and increased its binding affinity up to 1.9 × 10(-7) M in a module-by-module approach. The utility of the Fc-specific repebody was demonstrated by the performance of an immobilized repebody in affinity purification of antibodies from a mammalian cell-cultured medium. Bound-antibodies on an immobilized repebody were shown to be eluted at pH 4.0 with high purity (>94.6%) and recovery yield (>95.7%). The immobilized repebody allowed a repetitive purification process more than ten times without any loss of binding capability. The repebody remained almost intact even after incubation with 0.5 M NaOH for 15 days. The present approach could be effectively used for developing a repeat module-based binder for other target molecules for affinity purification.


Evidence that transcriptional activation by p53 plays a direct role in the induction of cellular senescence.
Wild-type p53 is necessary for the growth arrest of human diploid fibroblasts (HDF) (and many other cell types) at the end of their proliferative lifespan. Although p53 may actively mediate senescence, possibly in response to telomere erosion, it is however equally possible that it is merely a permissive factor required for response to some other inducer. To address this question, we have generated stable transfectants of early passage HDF, represented here by clone LacZ21, in which expression of a beta-galactosidase reporter construct reflects p53 transactivation activity. During continuous passage, the proportion of beta-gal positive LacZ21 cells remained below 2% for 25 population doublings (pd), first became significantly increased after 29 pd, and thereafter increased rapidly, reaching a maximum of 88% in fully-senescent cells (32 pd), which exceeded the response observed following an optimum dose (20 J/m2) of u.v. radiation. Correspondingly, the proportion of cells incorporating bromodeoxyuridine (BrdU) (initially 45-50%) began to fall at 29 pd and thereafter dropped rapidly to below 1% by pd 32. There was therefore a near-perfect reciprocal relationship between reporter construct expression and DNA synthesis as cells approached senescence. Furthermore, a dominant-negative p53 mutant (introduced by retroviral transduction) rescued LacZ21 cells from senescence and generated colonies with extended lifespan in which beta-gal expression was totally abolished. These data, although not excluding the need for other p53 functions, strongly suggest that p53-mediated transactivation of growth regulatory genes is a direct trigger, rather than a permissive factor, for cellular senescence.


Sexual behaviour, condom use, and disclosure of HIV status in HIV infected heterosexual individuals attending an inner London HIV clinic.
The National Strategy for Sexual Health and HIV for England (2001) emphasised the role of HIV services in reducing secondary transmission of HIV through prevention work with HIV infected people. To determine the sexual behaviour, condom use, and disclosure of HIV status of HIV infected heterosexuals attending an inner London HIV clinic. Cross sectional questionnaire study of heterosexual HIV infected individuals attending an HIV outpatient clinic. We collected demographic data for all respondents and sexual behaviour data for those sexually active over the past year using a self administered questionnaire. Viral load and CD4 count for responders and age, sex, ethnicity, viral load, and CD4 count for non-responders were obtained from the clinic database. The response rate was 47.3% (n = 142). 100 participants reported being sexually active in the past year, of whom 73% used condoms when they last had vaginal sex. Knowledge of partner's HIV status was the only variable significantly associated with the participant disclosing their HIV status to their partner (p<0.001). In those who had disclosed their status, only knowledge of partner's HIV status was significantly associated with condom use (p = 0.03). Issues relating to non-disclosure and partner notification in HIV infected heterosexuals will need to be better understood to improve sexual health in this group and to reduce onward transmission of HIV.


Amino acids in sheep production.
Increasing production efficiency with a high standard of animal welfare and respect for the environment is a goal of sheep farming systems. Substantial gains in productivity have been achieved through improved genetics, nutrition and management changes; however the survival and growth performance of multiple-born lambs still remains a problem. This is a significant production efficiency and animal well-being issue. There is a growing body of evidence that some amino acids have a role in regulating growth, reproduction and immunity through modulation of metabolic and cell signaling pathways. The purpose of this review is to provide an overview of what is currently known about the role of amino acids in sheep production and the potential for supplementation strategies to influence on-farm survival and growth of lambs.


Practical Advice for South Korean Medical Researchers Regarding Open-Access and Predatory Journals.
In recent decades, the volume of scholarly literature worldwide has increased significantly, and open-access publishing has become commonplace. These changes are even more dominant in South Korea. Comparing the periods of 1981-2000 and 2001-2020, the number of medical articles produced in Korea increased by 16.8 times on the Web of Science platform (13,223 to 222,771 papers). Before 1990, almost no open-access articles were produced in South Korea, but in the last 10 years open-access publications came to account for almost 40% of all South Korean publications on Web of Science. Along with the expansion of literature and the development of open-access publishing, predatory journals that seek profit without conducting quality assurance have appeared and undermined the academic corpus. In this rapidly changing environment, medical researchers have begun contemplating publication standards. In this article, recent trends in academic publishing are examined from international and South Korean perspectives, and the significance of open-access publishing and recent changes are discussed. Practical methods that can be used to select legitimate publishers, including open-access journals, and identify predatory journals are also discussed.


Pyogenic vertebral osteomyelitis.
I retrospectively reviewed the records of 111 patients who had pyogenic vertebral osteomyelitis unrelated to an open procedure on the spine. The mean age at the time of the diagnosis was sixty years (range, eighteen to eighty-four years); sixty-one patients (55 per cent) were sixty years old or more. Forty-four patients (40 per cent) had an impaired immune system secondary to diabetes mellitus, the use of corticosteroids, chemotherapy for cancer, rheumatic or immunological disease, renal or hepatic failure, malnutrition, or myelodysplasia. Magnetic resonance imaging, critical for the determination of an early diagnosis, was performed for 103 patients (93 per cent). The infection in sixty-eight patients (61 per cent) was diagnosed within one month after the onset of symptoms. The most frequent infecting organism was Staphylococcus aureus (forty patients; 36 per cent). The infection in forty-one patients (37 per cent) was caused by organisms, such as Staphylococcus epidermidis, Propionibacterium acnes, and diphtheroid species, that are traditionally considered to be of low virulence. The urinary tract was the most frequent source of infection (confirmed in thirteen patients and suspected in twenty-one). The success of non-operative treatment was predicted by four independent variables: an age of less than sixty years, the immune status, infection with Staphylococcus aureus, and a decreasing erythrocyte sedimentation rate. Forty-two patients were managed with debridement and arthrodesis. Fourteen of these patients also had instrumentation of the spine, in the presence of infection, without compromise of the outcome. Eighteen patients died by the time of the latest follow-up evaluation at a mean of four years (range, two years and two months to six years and six months): seven who had been managed non-operatively died in the first month after the diagnosis was made, three died in the acute postoperative period, three died of late complications of paraplegia, and five died of unrelated causes. None of the eighty-nine patients who were seen at a minimum of two years postoperatively had had late recurrence of infection. Chronic, severe back pain was noted in only seven patients.


Choroid plexus and cerebrospinal fluid production.
Acute progressive hydrocephalus occurred in 76 rhesus monkeys undergoing ventricular obstruction 2 to 6 months after choroid plexectomy and was only slightly less pronounced than in 73 nonplexectomized animals with similar ventricular obstructions. The composition of cerebrospinal fluid after choroid plexectomy was essentially normal. Experiments with ventricular perfusion techniques indicate only a slight reduction in cerebrospinal fluid production after choroid plexectomy. Thus, the choroid plexus is probably not the sole or even the major source of cerebrospinal fluid within the primate ventricular system.


Measuring Total Mercury Through Freeze-Thaw Cycles.
Total mercury (THg) analysis of fish tissue requires the samples to be frozen for long-term storage or analyzed immediately after the fish sample has been obtained. Such rigid field sampling protocols do not account for the logistical challenges that can occur if sampling is conducted in remote areas or collaboratively via community-based monitoring initiatives. Here we present a study that examines the loss of THg from Walleye tissue that has thawed for 3 and 5 days, mimicking potential thaw during transport from remote locations. THg concentrations were not significantly influenced (Kruskal-Wallace, p-value > 0.05) by the thaw and refreeze events. The mean THg concentration per day group was 0.67 ug/g, SD = 0.03, 0.65 ug/g, SD = 0.01, and 0.72 ug/g, SD = 0.01 for the control (no thaw), 3-day thaw and 5-day thaw, respectively. Results from this study suggest the analytical integrity of samples may be maintained if freeze-thaw situations arise when conducting community-based mercury monitoring programs.


Maternal transmission of a humanised Igf2r allele results in an Igf2 dependent hypomorphic and non-viable growth phenotype.
The cation independent mannose 6-phosphate/insulin-like growth factor 2 receptor (IGF2R) functions in the transportation and regulation of insulin-like growth factor 2 (IGF2) and mannose 6-phosphate modified proteins. The relative and specific titration of IGF2 by high affinity binding of IGF2R represents a mechanism that supports the parental conflict theory of genomic imprinting. Imprinting of Igf2 (paternal allele expressed) and Igf2r (maternal allele expressed) arose to regulate the relative supply of both proteins. Experiments in the mouse have established that loss of the maternal allele of Igf2r results in disproportionate growth and peri-natal lethality. In order to systematically investigate the consequences of loss of function and of hypomorphic alleles of Igf2r on growth functions, we introduced a conditional human IGF2R exon 3-48 cDNA into the intron 2 region of murine Igf2r. Here we show that the knock-in construct resulted in over-growth when the humanised Igf2r allele was maternally transmitted, a phenotype that was rescued by either paternal transmission of the humanised allele, expression of a wild-type paternal allele or loss of function of Igf2. We also show that expression of IGF2R protein was reduced to less than 50% overall in tissues previously known to be Igf2 growth dependent. This occurred despite the detection of mouse derived peptides, suggesting that trans-splicing of the knock-in human cDNA with the endogenous maternal mouse Igf2r allele. The phenotype following maternal transmission of the humanised allele resulted in overgrowth of the embryo, heart and placenta with partial peri-natal lethality, suggesting that further generation of hypomorphic Igf2r alleles are likely to be at the borderline of maintaining Igf2 dependent viability.


FOXM1/DVL2/Snail axis drives metastasis and chemoresistance of colorectal cancer.
Colorectal cancer (CRC) is the third most common type of cancer worldwide. Metastasis and chemoresistance are regarded as the two leading causes of treatment failure and high mortality in CRC. Forkhead Box M1 (FOXM1) has been involved in malignant behaviors of cancer. However, the role and mechanism of FOXM1 in simultaneously regulating metastasis and chemoresistance of CRC remain poorly understood. Here, we found that FOXM1 was overexpressed in oxaliplatin- and vincristine-resistant CRC cells (HCT-8/L-OHP and HCT-8/VCR) with enhanced metastatic potential, compared with HCT-8 cells. FOXM1 overexpression increased migration, invasion and drug-resistance to oxaliplatin and vincristine in HCT-8 cells, while FOXM1 knockdown using shFOXM1 impaired metastasis and drug-resistance in HCT-8/L-OHP and HCT-8/VCR cells. Moreover, FOXM1 up-regulated Snail to trigger epithelial-mesenchymal transition-like molecular changes and multidrug-resistance protein P-gp expression, while silencing Snail inhibited FOXM1-induced metastasis and drug-resistance. We further identified that disheveled-2 (DVL2) was crucial for FOXM1-induced Snail expression, metastasis and chemoresistance. Furthermore, FOXM1 bound to DVL2, and enhanced nuclear translocation of DVL2 and DVL2-mediated transcriptional activity of Wnt/β-catenin known to induce Snail expression. In conclusion, FOXM1/DVL2/Snail axis triggered aggressiveness of CRC. Blocking FOXM1/DVL2/Snail pathway simultaneously inhibited metastasis and chemoresistance in CRC cells, providing a new strategy for successful CRC treatment.


Adverse Economic Impact Associated With Blood Culture Contamination in a Pediatric Emergency Department.
Blood culture contamination (BCC) leads to unnecessary interventions including hospitalization, antibiotic administration and additional laboratory tests. Previous studies in adults revealed that BCC was associated with unnecessary financial expenditures. However, information pertaining to the pediatric population is limited. Therefore, we investigated the details of the adverse economic impact associated with BCC in a pediatric emergency department (ED) in Japan. This study was a retrospective, observational study. We collected data on blood cultures performed in patients 18 years of age in a pediatric ED. Medical records of patients with positive blood cultures were reviewed, and the information regarding adverse events related to BCC was extracted. Medical costs related to BCC were estimated from the data. In total, 13,139 sets of blood cultures were performed from April 2013 to June 2016, and 141 cases (1.1%) of BCC were identified. Among these, 106 patients (75%) experienced at least 1 adverse event associated with BCC. The total medical cost due to BCC was 4,076,713 Japanese yen. Multifaceted approaches targeting ED physicians including lectures on optimal blood collection methods and monthly feedback on BCC rates were effective in reducing the BCC rate and its related costs. Interventions associated with BCC were common and accounted for significant adverse economic impact on pediatric patients. Regular education and monitoring were effective in reducing BCC and its related costs.


Characterization of mutant human fibroblast cultures transformed with simian virus 40.
Fibroblast cell strains derived from a normal individual and from eight patients with various genetic mutations were transformed by a small-plaque variant of simian virus 40 (SV40, strain 776), cloned and studied after long-term in vitro maintenance. Seven of the cultures continued to express the mutant phenotype. Cultures derived from a patient with phosphoglycerate kinase I deficiency exhibited reappearance of normal enzyme activity after transformation. Compared to untransformed controls, all transformed cultures displayed decreased population doubling times, an increase in the relative number of cycling cells and increased saturation density on solid substrates, and did not show evidence of cellular senescence after long-term cultivation. Unlike previous studies on wild-type SV40-transformed human fibroblasts, the majority of cultures transformed by the small-plaque variant of SV40 did not exhibit signs of crisis. The cells also exhibited a decreased dependence on serum and were able to grow in semi-solid medium. The different transformed cultures expressed variable levels of SV40 large T-antigen, synthesized some infectious SV40 virus, and contained both unique arrangements and quantities of covalently integrated and episomal SV40 DNA. No correlation was observed between the rate of growth and synthesis of infectious virus in the different transformed clones. These studies indicate that this small-plaque variant of SV40 can be used effectively to generate long-lived human cultures, which generally retain their mutant phenotype. Transformation with this SV40 variant permits the generation of large quantities of clonal cell cultures for the biochemical and molecular analysis of their genetic defects.


Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies.
Neutralizing monoclonal antibodies (MAbs) were produced in BALB/c mice immunized with live modified vaccinia virus Ankara or infected with sublethal doses of the neurovirulent vaccinia virus strain Munich 1. The immunization scheme proved to be important for obtaining MAbs of different specificity. The MAbs could be classified into three epitope groups (1 A, 1 B and 2). Immunogold electron microscopy demonstrated that the epitopes were localized on the virus surface. In immunoblotting, MAbs were reactive with polypeptides of 14K, 16K and 30K. Purified MAbs binding to the epitopes 1 A and 2 showed a 50% reduction of 100 p.f.u./0.05 ml vaccinia virus M1 with respectively 3.9 and 5.9 ng of immunoglobulin/0.05 ml. MAbs binding to the epitope 1 B neutralized the virus at a concentration of 250 ng/0.05 ml. In intraperitoneal challenge experiments, MAbs binding to the epitopes 1 A and 2 protected mice against 4 LD50 of vaccinia virus M1, but not against local lesions by subcutaneous application. MAbs against epitope 1 B had no protective effect in vivo. The three epitopes were present in 14 of 16 orthopoxviruses tested but with quantitative differences. Maximal binding (Vmax) and the antibody concentration at half-maximal binding (Km) which were calculated as for Michaelis-Menten kinetics from regression analysis of the ELISA data and the MAb concentration giving 50% plaque reduction were the basis for the evaluation. In monkey-pox virus Kopenhagen the epitopes 1 A and 1 B were absent. MAbs binding to epitope 2 reacted just as well as with vaccinia viruses. Ectromelia virus lacked all the epitopes.


Unsafe and violent behavior in commercials aired during televised major sporting events.
Injuries are the leading cause of death in children, and media exposure seems to increase children's risk-taking behavior. Televised sports are commonly viewed by children. The objective of this study was to determine the proportion of commercials that depict violence or other unsafe behavior during major televised sporting events that are aired before 9:00 pm. We obtained a list of the 50 sports programs that were most highly rated by Nielsen Media Research and that were televised between September 1, 2001, and September 1, 2002. These 50 programs included Winter Olympics events (n = 15), National Football League (NFL) regular season games (n = 14), NFL playoff games (n = 10), Major League Baseball World Series and playoff games (n = 7), the NFL Super Bowl (n = 1), the National Basketball Association Western Conference Final Game (n = 1), the College Football Rose Bowl (n = 1), and the National Collegiate Athletic Association Basketball Championship game (n = 1). Two other events were reviewed as well: the final round of the Masters Golf Championship, because it was the only sporting event rated in the top 50 of the previous year that was not represented by a similar sporting event in the study year, and the Daytona 500 National Association for Stock Car Auto Racing race, because it was the only event rated among the top 75 of the study year that was not represented by a similar event (ie, there were no other golfing or auto racing events reviewed). These events were included because different sporting events may attract different viewers and different advertisements; thus, their inclusion provides a more comprehensive evaluation of the topic. For sporting events with >3 programs in the top 50 (NFL regular season games, NFL playoff games, Winter Olympic events, and Major League Baseball World Series), representative samples of events were assessed. Surrogate events were analyzed for programs that were aired after 9:00 PM (Eastern Time) to control for the reduced likelihood of viewing by children after 9:00 PM. For example, afternoon telecasts of the National Collegiate Athletic Association Regional Final and National Semifinal games were assessed in place of the Championship game, which started after 9:00 PM. Weekend afternoon telecasts of the Winter Olympics (3 successive weekends) were assessed as surrogates for all Olympics telecasts. For the World Series, the Super Bowl, and the National Basketball Association Conference Championship games, which began before 9:00 PM but ended after that time, we evaluated only commercials that aired before 9:00 PM. All commercials that were aired during these programs were reviewed at standard speed for unsafe behavior or violence. Commercials that were aired during pregame, postgame, or halftime periods were not assessed. Commercials were categorized according to the product being advertised. Unsafe behavior was simply defined as any action that could have harmful consequences or that contravened the injury prevention recommendations of national organizations. Violence was defined as any intentional physical contact by an aggressor that had the potential to inflict injury or harm or the legitimate threat of such action. All commercials were reviewed by at least 2 investigators; when the 2 could not agree on the findings, a third investigator was used to resolve differences. The percentage of commercials and of commercial breaks that portrayed violent or unsafe behavior was determined for each category of event and advertised product. A commercial break was defined as a series of commercials shown during a single break from the sporting event. chi2 analysis was used for all analyses, and relative risks with 95% confidence intervals were determined. The proportion of commercials that depicted unsafe behavior and/or violence during each sporting event was compared with the proportion of such commercials that were observed during the Masters Golf Tournament (which had the lowest proportion of commercials depicting such behavior). The proportion of commercials that contained violent/unsafe behavior for each advertised product was compared with the proportion of such commercials that advertised food or nonalcoholic beverages. Food and nonalcoholic beverage commercials were selected as the reference because they are a well-defined, common category of commercial. Of the 1185 commercials assessed, 14% (n = 165) displayed unsafe behavior and 6% (n = 66) depicted violence. Of the 322 commercial breaks, 158 (49%) contained at least 1 commercial showing unsafe behavior or violence. Sixty-three commercials required review by a third investigator to adjudicate violence or unsafe behavior; 20 of 52 were ultimately judged to portray unsafe behavior, and 4 of 11 were ultimately judged to portray violence. Sporting events differed in the proportion of commercials that showed violence or unsafe behavior. The Super Bowl had the highest proportion of such commercials, and the Masters Golf Tournament had the least (relative risk: 4.3; 95% confidence interval: 1.4-12.5). The Masters Golf Tournament was noteworthy for the complete absence of violent commercials. Only 18% of reviewed commercials advertised movies or television programs, yet these commercials accounted for 86% of all violent commercials. Forty-eight percent of commercials that contained violence were for movies, and an additional 38% were for television programs. Nearly two thirds of all commercials for movies contained violence, whereas 15% of all commercials for television programs contained violence, a rate that increased to 22% when commercials for other sporting events were excluded. Several categories of commercials portrayed unsafe behaviors; commercials for automobiles accounted for the most. In 8 different categories, 10% or more of the commercials depicted unsafe behavior, and 7 were significantly more likely to depict such behavior than a food or beverage commercial. Children who watch televised sports view a significant amount of violent and unsafe behavior. In accordance with American Academy of Pediatrics recommendations for television viewing, parents should both limit and directly supervise children's viewing of these events. Our findings suggest that parents should remain present during commercials or should consider implementing commercial-skipping technology. In addition, efforts should be made to regulate the content of commercials that promote television programs and movies on the basis of the hour at which the sporting event is aired. Moreover, the sports, movie, and television industries should be encouraged to adopt models of advertising that limit or eliminate such content. These efforts could help to ensure that the viewing of televised sporting events is a safe and positive experience for children.


The comparison of latent variable models of non-psychotic psychiatric morbidity in four culturally diverse populations.
Factor analysis has been employed to identify latent variables that are unifying constructs and that parsimoniously describe correlations among a related group of variables. Confirmatory factor analysis is used to test hypothesized factor structures for a set of variables; it can also, as in this paper be used to model data from two or more groups simultaneously to determine whether they have the same factor structure. Non-psychotic psychiatric morbidity, elicited by the Revised Clinical Interview Schedule (CIS-R), from four culturally diverse populations was compared. Confirmatory factor analysis was employed to compare the factor structures of CIS-R data sets from Santiago, Harare, Rotherhithe and Ealing. These structures were compared with hypothetical one and two factor (depression-anxiety) models. The models fitted well with the different data sets. The depression-anxiety model was marginally superior to the one factor model as judged by various statistical measures of fit. The two factors in depression-anxiety model were, however, highly correlated. The findings suggest that symptoms of emotional distress seem to have the same factor structure across cultures.


Direct silica coating of drug crystals for ultra-high loading.
To push the limit of synthetic control, we create a thin layer (5 nm) of silica on the surface of drug nanocrystals, achieving a loading content (88%) that approaches the theoretical limit. The uniform silica shell provides a tailored diffusion barrier for controlled drug release. The method can be generally applied to 11 organic crystals, including 4 drugs.


Functional assay for BRCA1: mutagenesis of the COOH-terminal region reveals critical residues for transcription activation.
The breast and ovarian cancer susceptibility gene product BRCA1 is a tumor suppressor, but its precise biochemical function remains unknown. The BRCA1 COOH terminus acts as a transcription activation domain, and germ-line cancer- predisposing mutations in this region abolish transcription activation, whereas benign polymorphisms do not. These results raise the possibility that loss of transcription activation by BRCA1 is crucial for oncogenesis. Therefore, identification of residues involved in transcription activation by BRCA1 will help understand why particular germ-line missense mutations are deleterious and may provide more reliable presymptomatic risk assessment. The BRCA1 COOH terminus (amino acids 1560-1863) consists of two BRCTs preceded by a region likely to be nonglobular. We combined site-directed and random mutagenesis, followed by a functional transcription assay in yeast: (a) error-prone PCR-induced random mutagenesis generated eight unique missense mutations causing loss of function, six of which targeted hydrophobic residues conserved in canine, mouse, rat, and human BRCA1; (b) random insertion of a variable pentapeptide cassette generated 21 insertion mutants. All pentapeptide insertions NH2-terminal to the BRCTs retained wild-type activity, whereas insertions in the BRCTs were, with few exceptions, deleterious; and (c) site-directed mutagenesis was used to characterize five known germ-line mutations and to perform deletion analysis of the COOH terminus. Deletion analysis revealed that the integrity of the most COOH-terminal hydrophobic cluster (I1855, L1854, and Y1853) is necessary for activity. We conclude that the integrity of the BRCT domains is crucial for transcription activation and that hydrophobic residues may be important for BRCT function. Therefore, the yeast-based assay for transcription activation can be used successfully to provide tools for structure-function analysis of BRCA1 and may form the basis of a BRCA1 functional assay.


Development of a model system for rapid assessment of insect mortality in heated controlled atmosphere quarantine treatments.
The development of postharvest quarantine treatments can be both expensive and time-consuming. It is necessary to determine the species and stage of the pest most tolerant to the treatment, if more than one species is the target of the treatment. Initial laboratory studies often include infesting the commodity with various egg and larval stages of the pest and performing treatments and evaluations of the fruit. In collaboration with others, I have previously developed combination high temperature under controlled atmosphere treatments against two quarantine pests in apples (Malus spp.) and peaches and nectarines (both Prunus spp.). I decided to develop an artificial system that can be used for these initial tests without the need for infesting large quantities of the fruit. I tested the system on the immature stages of the pests under regular air and controlled atmospheres by using the controlled atmosphere water bath system. This system can be used for rapid assessment of the most tolerant stage and species of a pest to a combination heat and controlled atmosphere treatment without the expense of infesting, treating, and evaluating the commodity.


Quantitative competitive PCR as a technique for exploring flea-Yersina pestis dynamics.
We used a quantitative competitive polymerase chain reaction assay to quantify Yersinia pestis loads in fleas and bacteremia levels in mice that were used as sources of infectious blood meals for feeding the fleas. Xenopsylla cheopis, the Oriental rat flea, achieved higher infection rates, developed greater bacterial loads, and became infectious more rapidly than Oropsylla montana, a ground squirrel flea. Both flea species required about 10(6) Y. pestis cells per flea to be able to transmit to mice. Most fleas that achieved these levels, however, were incapable of transmitting. Our results suggest that at the time of flea feeding, host blood must contain > or = 10(6) bacteria/ml to result in detectable Y. pestis infections in these fleas, and > or = 10(7) bacteria/mL to cause infection levels sufficient for both species to eventually become capable of transmitting Y. pestis to uninfected mice. Yersinia pestis colonies primarily developed in the midguts of O. montana, whereas infections in X. cheopis often developed simultaneously in the proventriculus and the midgut. These findings were visually confirmed by infecting fleas with a strain of Y. pestis that had been transformed with the green fluorescent protein gene.


Upregulation of soluble and membrane-bound human leukocyte antigen G expression is primarily observed in the milder histopathological stages of chronic hepatitis C virus infection.
Chronic hepatitis C virus (HCV) infection is a worldwide health problem that may evolve to cirrhosis and hepatocellular carcinoma. Incompletely understood immune system mechanisms have been associated with impaired viral clearance. The nonclassical class I human leukocyte antigen G (HLA-G) molecule may downregulate immune system cell functions exhibiting well-recognized tolerogenic properties. HCV genotype was analyzed in chronic HCV-infected patients. Because HLA-G expression may be induced by certain viruses, we evaluated the presence of HLA-G in the liver microenvironment obtained from 89 biopsies of patients harboring chronic HCV infection and stratified according to clinical and histopathological features. Overall, data indicated that HCV genotype 1 was predominant, especially subgenotype 1a, with a prevalence of 87%. HLA-G expression was observed in 45 (51%) liver specimens, and it was more frequent in milder stages of chronic hepatitis (67.4%) than in moderate (27.8%; p = 0.009) and severe (36.0%; p = 0.021) stages of the disease. Altogether, these results suggest that the expression of HLA-G in the context of HCV is a complex process modulated by many factors, which may contribute to an immunologic environment favoring viral persistence. However, because the milder forms predominantly expressed HLA-G, a protective role of this molecule may not be excluded.


Expression pattern in the antennae of a newly isolated lepidopteran Gq protein alpha subunit cDNA.
From the antennae of the moth Mamestra brassicae, we have identified a lepidopteran G protein alpha subunit belonging to the Gq family, through immunological detection in crude antennal extract and antennal primary cell cultures, followed by molecular cloning. The complete cDNA sequence (1540 bp) contains an open reading frame encoding a protein of 353 amino acids. This deduced sequence possesses all of the characteristics of the Gq family and shares a very high degree of amino-acid sequence identity with vertebrate (80% with mouse or human Gqalpha) and invertebrate subunits (varying between 60 and 87% for Gqalpha from organisms as diverse as sponge and Drosophila). The expression pattern of the Gq subunit in adult antennae was associated with the olfactory sensilla suggesting a specific role in olfaction. These data provide molecular evidence for a component of the phosphoinositide signaling pathway in moth antennae: this G protein alpha subunit may be involved in the olfaction transduction process through interaction with G-protein-coupled receptors, stimulating the phospholipase C mediated second messenger pathway.


Patient-Specific Variations in Local Strain Patterns on the Surface of a Trussed Titanium Interbody Cage.
Introduction: 3D printed trussed titanium interbody cages may deliver bone stimulating mechanobiological strains to cells attached at their surface. The exact size and distribution of these strains may depend on patient-specific factors, but the influence of these factors remains unknown. Therefore, this study aimed to determine patient-specific variations in local strain patterns on the surface of a trussed titanium interbody fusion cage. Materials and Methods: Four patients eligible for spinal fusion surgery with the same cage size were selected from a larger database. For these cases, patient-specific finite element models of the lumbar spine including the same trussed titanium cage were made. Functional dynamics of the non-operated lumbar spinal segments, as well as local cage strains and caudal endplate stresses at the operated segment, were evaluated under physiological extension/flexion movement of the lumbar spine. Results: All patient-specific models revealed physiologically realistic functional dynamics of the operated spine. In all patients, approximately 30% of the total cage surface experienced strain values relevant for preserving bone homeostasis and stimulating bone formation. Mean caudal endplate contact pressures varied up to 10 MPa. Both surface strains and endplate contact pressures varied more between loading conditions than between patients. Conclusions: This study demonstrates the applicability of patient-specific finite element models to quantify the impact of patient-specific factors such as bone density, degenerative state of the spine, and spinal curvature on interbody cage loading. In the future, the same framework might be further developed in order to establish a pipeline for interbody cage design optimizations.


[The nature of the influenza C virus receptor and the specificity of the receptor-destroying enzyme].
Bacterial neuraminidases destroy influenza C virus receptors of chick erythrocytes and inactivate hemagglutination inhibitors: rat alpha 1-macroglobulin (RMG) and bovine submaxillary mucin (BSM). These data indicate that neuraminic acid may be a component of influenza C virus receptor. The inhibiting activity of RMG and BSM is also eliminated by the receptor-destroying enzyme (RDE) of influenza C virus. After inactivation, the inhibitors (RMG and BSM) contain a reduced amount of N-acetyl-9-0-acetylneuraminic acid (Neu5, 9Ac2) and a larger amount of N-acetylneuraminic acid (Neu5 Ac). Transformation of Neu5, 9Ac2 into Neu5 Ac may also occur upon incubation of free neuraminic acid with influenza C virus. These data indicate that the RDE of influenza C virus is neuraminate-O-acetylesterase (N-acyl-9 4-O-acetylneuraminate O-acetylhydrolase (EC 22.214.171.124). It was shown that inhibition of influenza C virus hemagglutination by RMG and BSM and, apparently, adhesion of the virus to the cell surface involves binding of influenza C virus with Neu5, 9Ac2.


The role of the natural epidemic dynamics and migration in explaining the course of the HIV epidemic in rural Uganda: a modelling study.
Human immunodeficiency virus (HIV) prevalence in Uganda fell during the 1990s and has risen since 2000. The changing trends since 2000 may be due to changing risky sexual behaviour, but other factors may also contribute. We explore the possible impact that two factors may have on cohort trends: natural epidemic dynamics and local migration. We simultaneously fit a mathematical model to the contrasting prevalence trends by age/gender in a southwest Ugandan cohort, which has never been done before. We fit a model to HIV trends, by age groups and gender, assuming: (i) neither migration nor intentional behaviour change, (ii) migration changes, (iii) intentional behaviour changes and (iv) both change. The model fits were assessed through sum of squares goodness of fits. We also explored the impact of the natural course of the epidemic on average partner acquisition rates, under the condition of no intentional behaviour change over time. Without allowing intentional change in partner acquisition rates, an excellent fit was achievable to prevalence trends by gender. However, all of the contrasting trends by age/gender could not be replicated simultaneously. Adding intentional behaviour change improved the fit to some groups, but not all. Adding both intentional behaviour change and migration further improved the fit. While some of the increasing HIV prevalence in Uganda since 2000 may be due to increased risky behaviour, some of the observed epidemiologic trends would likely occur without any intentional change in behaviour. Average population-level behaviour can change due to preferential mortality among higher risk takers, without individuals changing their behaviour.