Abundant Monovalent Ions as Environmental Signposts for Pathogens during Host Colonization.
Host colonization by a pathogen requires proper sensing and response to local environmental cues, to ensure adaptation and continued survival within the host. The ionic milieu represents a critical potential source of environmental cues, and indeed, there has been extensive study of the interplay between host and pathogen in the context of metals such as iron, zinc, and manganese, vital ions that are actively sequestered by the host. The inherent non-uniformity of the ionic milieu also extends, however, to "abundant" ions such as chloride and potassium, whose concentrations vary greatly between tissue and cellular locations, and with the immune response. Despite this, the concept of abundant ions as environmental cues and key players in host-pathogen interactions is only just emerging. Focusing on chloride and potassium, this review brings together studies across multiple bacterial and parasitic species that have begun to define both how these abundant ions are exploited as cues during host infection, and how they can be actively manipulated by pathogens during host colonization. The close links between ion homeostasis and sensing/response to different ionic signals, and the importance of studying pathogen response to cues in combination, are also discussed, while considering the fundamental insight still to be uncovered from further studies in this nascent area of inquiry.


The genetic basis of speciation in the Giliopsis lineage of Ipomopsis (Polemoniaceae).
One of the most powerful drivers of speciation in plants is pollinator-mediated disruptive selection, which leads to the divergence of floral traits adapted to the morphology and behavior of different pollinators. Despite the widespread importance of this speciation mechanism, its genetic basis has been explored in only a few groups. Here, we characterize the genetic basis of pollinator-mediated divergence of two species in genus Ipomopsis, I. guttata and I. tenuifolia, using quantitative trait locus (QTL) analyses of floral traits and other variable phenotypes. We detected one to six QTLs per trait, with each QTL generally explaining small to modest amounts of the phenotypic variance of a backcross hybrid population. In contrast, flowering time and anthocyanin abundance (a metric of color variation) were controlled by a few QTLs of relatively large effect. QTLs were strongly clustered within linkage groups, with 26 of 37 QTLs localized to six marker-interval 'hotspots,' all of which harbored pleiotropic QTLs. In contrast to other studies that have examined the genetic basis of pollinator shifts, our results indicate that, in general, mutations of small to modest effect on phenotype were involved. Thus, the evolutionary transition between the distinct pollination modes of I. guttata and I. tenuifolia likely proceeded incrementally, rather than saltationally.


Importance of bulk states for the electronic structure of semiconductor surfaces: implications for finite slabs.
We investigate the influence of slab thickness on the electronic structure of the Si(1 0 0)- p([Formula: see text]) surface in density functional theory (DFT) calculations, considering both density of states and band structure. Our calculations, with slab thicknesses of up to 78 atomic layers, reveal that the slab thickness profoundly affects the surface band structure, particularly the dangling bond states of the silicon dimers near the Fermi level. We find that, to precisely reproduce the surface bands, the slab thickness needs to be large enough to completely converge the bulk bands in the slab. In the case of the Si(1 0 0) surface, the dispersion features of the surface bands, such as the band shape and width, converge when the slab thickness is larger than 30 layers. Complete convergence of both the surface and bulk bands in the slab is only achieved when the slab thickness is greater than 60 layers.


Retinal pigment epithelial integrity is compromised in the developing albino mouse retina.
In the developing murine eye, melanin synthesis in the retinal pigment epithelium (RPE) coincides with neurogenesis of retinal ganglion cells (RGCs). Disruption of pigmentation in the albino RPE is associated with delayed neurogenesis in the ventrotemporal retina, the source of ipsilateral RGCs, and a reduced ipsilateral RGC projection. To begin to unravel how melanogenesis and the RPE regulate RGC neurogenesis and cell subpopulation specification, we compared the features of albino and pigmented mouse RPE cells during the period of RGC neurogenesis (embryonic day, E, 12.5 to 18.5) when the RPE is closely apposed to developing RGC precursors. At E12.5 and E15.5, although albino and pigmented RPE cells express RPE markers Otx2 and Mitf similarly, albino RPE cells are irregularly shaped and have fewer melanosomes compared with pigmented RPE cells. The adherens junction protein P-cadherin appears loosely distributed within the albino RPE cells rather than tightly localized on the cell membrane, as in pigmented RPE. Connexin 43 (gap junction protein) is expressed in pigmented and albino RPE cells at E13.5 but at E15.5 albino RPE cells have fewer small connexin 43 puncta, and a larger fraction of phosphorylated connexin 43 at serine 368. These results suggest that the lack of pigment in the RPE results in impaired RPE cell integrity and communication via gap junctions between RPE and neural retina during RGC neurogenesis. Our findings should pave the way for further investigation of the role of RPE in regulating RGC development toward achieving proper RGC axon decussation. J. Comp. Neurol. 524:3696-3716, 2016. © 2016 Wiley Periodicals, Inc.


Distribution of Cellular Carbohydrates during Development of the Mycelium and Fruitbodies of Flammulina velutipes.
Flammulina velutipes (Curt. ex Fr.) Sing. was grown on potato-glucose solution freed of most starch. Glucose uptake and dry weight changes in the colony indicated that the large fruitbodies derived their substrates partly from glucose remaining in the medium and partly from cellular constituents stored in the mycelium and small fruitbodies. Changes in the amounts of low molecular weight carbohydrates, glycogen, and four cell wall polysaccharide fractions were followed in the mycelium and fruitbodies. Trehalose, arabitol, and smaller amounts of mannitol were the main stored low molecular weight carbohydrates. A large net loss of these compounds occurred in the mycelium and small fruitbodies after their growth ceased. The carbohydrates accumulated in the large fruitbodies, but were also partly metabolized in the colony. Reducing sugars were minor components, and included about 30 to 50% glucose and a small undetermined quantity of fructose. Glycogen was the main storage carbohydrate in the mycelium, and was also stored in the small fruitbodies. It was broken down in both structures during growth of the large fruitbodies which accumulated only small amounts. During the same period, almost 45% of the maximum amount of cell wall polysaccharides were degraded in the small fruitbodies, but not in the mycelium.By feeding (14)C-glucose in replacement medium, incorporation of radioactivity into carbohydrates was followed in the colony during fruit-body development. Total incorporation was highest in trehalose, next highest in glycogen, and the rest was found in polyols and cell wall polysaccharides except for a few per cent which remained in endogenous glucose. In the large fruitbodies, specific radioactivity in glucose was much lower than in trehalose and mannitol. The labeling patterns in the mycelium and large fruitbodies suggested that trehalose, mannitol, and possibly arabitol were translocated into the stipes and pilei.


A simple and versatile one-pot synthesis of meso-substituted trans-A2B-corroles.
We have developed a new methodology that affords regioisomerically pure trans-A2B-corroles. The corrole formation reaction involves the acid-catalyzed condensation of a dipyrromethane and an aldehyde followed by oxidation with DDQ. Optimal conditions for the condensation were identified after examining various reaction parameters (solvent, acid, concentration, time). The conditions identified (CH2Cl2, [DPM] = 33 mM, [aldehyde] = 17 mM, [TFA] = 1.3 mM (for sterically hindered DPMs) or [TFA] = 0.26 mM (for sterically unhindered DPMs), 5 h, room temperature) resulted in the formation of corroles in 3-25% yield without detectable scrambling. The synthesis is compatible with diverse functionalities: ester, nitrile, ether, fluoro, hydroxy, iodo, nitro, thioacetate, methylsulfoxy. In total 21 corroles of type trans-A2B were prepared. Three exemplary corrole syntheses were successfully carried out at 8 mmol scale. Corroles 23, 30, and 41 (160-600 mg) were obtained in essentially the same yield as in small scale experiments.


Outcomes of surgery in the Medicare aged population: rehospitalization after surgery.
Using 1979 and 1980 data, rehospitalization rates following eight common surgical procedures are examined for aged Medicare beneficiaries. Rehospitalization rates within 30 days after discharge from the surgical stay varied considerably among procedures and were higher for older beneficiaries. Patients residing in the Northeast had the lowest rates of rehospitalization, although their rehospitalizations tended to be lengthier than those elsewhere. Rehospitalization rates were also tracked for 9 months following discharge from the surgical stay. For all procedures, rehospitalization rates decreased during the 9 months after discharge, but they remained above the prevailing hospitalization rate for the Medicare aged population for the entire 9 months. Principal diagnoses associated with rehospitalizations within 30 days were often related to the body system on which surgery was initially performed, suggesting that many rehospitalizations are for continuing problems related to the initial condition that necessitated surgery. Rehospitalization rates presented in this article will serve as baseline data for monitoring trends under medicare's prospective payment system for hospitals.


Patient safety culture and leadership within Canada's Academic Health Science Centres: towards the development of a collaborative position paper.
Currently, the Academy of Canadian Executive Nurses (ACEN) is working with the Association of Canadian Academic Healthcare Organizations (ACAHO) to develop a joint position paper on patient safety cultures and leadership within Academic Health Science Centres (AHSCs). Pressures to improve patient safety within our healthcare system are gaining momentum daily. Because AHSCs in Canada are the key organizations that are positioned regionally and nationally, where service delivery is the platform for the education of future healthcare providers, and where the development of new knowledge and innovation through research occurs, leadership for patient safety logically must emanate from them. As a primer, ACEN provides an overview of current patient safety initiatives in AHSCs to date. In addition, the following six key areas for action are identified to ensure that AHSCs continue to be leaders in delivering quality, safe healthcare in Canada. These include: (1) strategic orientation to safety culture and quality improvement, (2) open and transparent disclosure policies, (3) health human resources integral to ensuring patient safety practices, (4) effective linkages between AHSCs and academic institutions, (5) national patient safety accountability initiatives and (6) collaborative team practice.


Quality Metrics in Neonatal and Pediatric Critical Care Transport: A National Delphi Project.
The transport of neonatal and pediatric patients to tertiary care facilities for specialized care demands monitoring the quality of care delivered during transport and its impact on patient outcomes. In 2011, pediatric transport teams in Ohio met to identify quality indicators permitting comparisons among programs. However, no set of national consensus quality metrics exists for benchmarking transport teams. The aim of this project was to achieve national consensus on appropriate neonatal and pediatric transport quality metrics. Modified Delphi technique. The first round of consensus determination was via electronic mail survey, followed by rounds of consensus determination in-person at the American Academy of Pediatrics Section on Transport Medicine's 2012 Quality Metrics Summit. All attendees of the American Academy of Pediatrics Section on Transport Medicine Quality Metrics Summit, conducted on October 21-23, 2012, in New Orleans, LA, were eligible to participate. Candidate quality metrics were identified through literature review and those metrics currently tracked by participating programs. Participants were asked in a series of rounds to identify "very important" quality metrics for transport. It was determined a priori that consensus on a metric's importance was achieved when at least 70% of respondents were in agreement. This is consistent with other Delphi studies. Eighty-two candidate metrics were considered initially. Ultimately, 12 metrics achieved consensus as "very important" to transport. These include metrics related to airway management, team mobilization time, patient and crew injuries, and adverse patient care events. Definitions were assigned to the 12 metrics to facilitate uniform data tracking among programs. The authors succeeded in achieving consensus among a diverse group of national transport experts on 12 core neonatal and pediatric transport quality metrics. We propose that transport teams across the country use these metrics to benchmark and guide their quality improvement activities.


Computing the compliance of physician drug orders with guidelines using an OWL2 reasoner and standard drug resources.
Assessing the conformity of a physician's prescription to a given recommended prescription is not obvious since both prescriptions are expressed at different levels of abstraction and may concern only a subpart of the whole order. Recent formalisms (OWL2) and tools (reasoners) from the semantic web technologies are becoming available to represent defined concepts and to handle classification services. We propose a generic framework based on such technologies, using available standardized drug resources, to compute the compliance of a given drug order to a recommended prescription, such that the subsumption relationship yields the conformity relationship between the order and the recommendation. The ATC drug classification has been used as a local ontology. The method has been successfully implemented for arterial hypertension management for which we had a sample of antihypertensive orders. However, supplemental standardized drug knowledge is needed to correctly compare drug orders to recommended orders.


[Clinical study on single-dose rhG-CSF combined with chemotherapy in autologous peripheral blood stem cell transplantation].
This study was designed to evaluate the effectiveness of high-dose chemotherapy plus single daily dose of 250 micrograms/d rhG-CSF(lenograstim) in autologous peripheral blood stem cells mobilization and hematopoietic reconstitution after transplantation. The study included a group of 55 consecutive patients with hematological malignancies (28 cases of acute myelogenous leukemia, 9 cases of acute lymphoblastic leukemia, 14 cases of non-Hodgkin's lymphoma, 4 cases of other disease) from October 1999 to December 2001 undergoing either EA or other intensive chemotherapies as CTX, G-CSF mobilization began from the day when the WBC count reached nadir until the day of apheresis, then after stem cell reinfusion G-CSF support was given on day +3 until neutrophil recovery. The dose of G-CSF both in mobilization and recovery support was 250 micrograms/d s.c. despite the body weight of patients. The average time of G-CSF administrating as a mobilizing reagent was 7.12 days(range 3-14 days), and the mean time of interval from the beginning of chemotherapy to apheresis was 18.08 +/- 3.63 days. All patients need an average of 1.3 times for apheresis. The median mononuclear cell and CD34+ cell yield were 4.09 x 10(8)/kg and 8.5 x 10(6)/kg, respectively. G-CSF was used after transplantation for a median time of 8 days(range 4-19). All patients achieved an ANC > 0.5 x 10(9)/L after a median of 10.5 days (range 8-26). No severe toxic reaction was observed. We concluded that chemotherapy plus rhG-CSF(lenograstim) given in the single dose fashion can be used in both mobilizing and post transplantation setting with satisfying outcome.


The roles of motilin and ghrelin in gastrointestinal motility.
In structure, ghrelin resembles motilin. The two peptides are considered to be members of the motilin-ghrelin peptide family. Motilin is considered to be an endocrine regulator of the interdigestive migrating contractions, the fasted motor pattern in the gastrointestinal (GI) tract. It has been reported that ghrelin stimulates GI motility. The gastrokinetic capacity of ghrelin has been well documented in the rodent. However, there have been few positive reports of the gastrokinetic capacity of ghrelin in dogs. Some reports with human subjects have shown that an i.v. ghrelin injection accelerated gastric emptying of a meal and improved meal-related symptoms. These results suggest that ghrelin has potential as a prokinetic. However, it seems unlikely that plasma ghrelin would play a physiological role in these digestive physiological events and stimulate gastric emptying, as these outcomes would appear to be in contradiction with the suppression of the endogenous release of ghrelin after eating. The physiological roles of ghrelin need to be clarified.


Hepatic metastases: in vivo assessment of perfusion parameters at dynamic contrast-enhanced MR imaging with dual-input two-compartment tracer kinetics model.
This study was institutional review board approved, with waived patient consent for retrospective analysis of the data. The hepatic perfusion at dynamic contrast material-enhanced magnetic resonance (MR) imaging was commonly described and assessed by using a dual-input one-compartment tracer kinetics model. Although the tracer kinetics in normal liver parenchyma can be described by using a single compartment, functional changes in the tumor microenvironment can result in distinctly different tracer behavior that entails a second tissue compartment. A dual-input two-compartment model is proposed to describe the tracer behavior in hepatic metastases. The authors applied this model to the dynamic MR imaging data obtained in three patients. Perfusion parameter maps and region-of-interest analysis revealed that tracer behavior in hepatic metastases-in contrast to that in surrounding normal liver tissue, which effectively involves one compartment-can be described by using two compartments.


Tissue-specific stilbene accumulation is an early response to wounding/grafting as revealed by using spatial and temporal metabolomics.
Grafting is widely used in horticulture. Shortly after grafting, callus tissues appear at the graft interface and the vascular tissues of the scion and rootstock connect. The graft interface contains a complex mix of tissues, we hypothesised that each tissue has its own metabolic response to wounding/grafting and accumulates different metabolites at different rates. We made intact and wounded cuttings and grafts of grapevine, and then measured changes in bulk flavonoid, phenolic acid and stilbenoid concentration and used metabolite imaging to study tissue-specific responses. We show that some metabolites rapidly accumulate in specific tissues after grafting, for example, stilbene monomers accumulate in necrotic tissues surrounding mature xylem vessels. Whereas other metabolites, such as complex stilbenes, accumulate in the same tissues at later stages. We also observe that other metabolites accumulate in the newly formed callus tissue and identify genotype-specific responses. In addition, exogenous resveratrol application did not modify grafting success rate, potentially suggesting that the accumulation of resveratrol at the graft interface is not linked to graft union formation. The increasing concentration of complex stilbenes often occurs in response to plant stresses (via unknown mechanisms), and potentially increases antioxidant activity and antifungal capacities.


Allograft tracheoplasty technique for management of refractory tracheal stenosis.
Extensive tracheal airway defects represent a clinical dilemma. Although resection and reanastomosis and staged tracheoplasty may prove beneficial in some cases, recurrent or extensive circumferential stenosis remains a reconstructive challenge. We report the use of the allograft tracheoplasty technique for the reconstruction of recurrent, extensive defects of the trachea and cricoid. Nine consecutive patients with recurrent tracheal stenosis were treated with the two-stage allograft tracheoplasty technique. A retrospective review was performed to evaluate for prior surgery, length of stenosis, surgical technique, and outcome. All 9 patients underwent multiple surgical procedures for acquired tracheal stenosis (average, 3.4 procedures) before undergoing the allograft tracheoplasty technique. Before surgery, all patients were tracheotomy-dependent. The patients were assessed 8 to 39 months after allograft tracheoplasty. The primary airway disorders included postintubation stenosis (n = 6), surgical resection for malignancy (n = 1), and idiopathic stenosis (n = 2). Three defects involved 30% to 60% of the cricoid cartilage, and 4 defects were complete circumferential tracheal defects. Five patients underwent an island deltopectoral flap for closure of the tracheoplasty site. One patient had a superficial wound infection at the cartilage recipient site, and 1 patient had a hematoma at the deltopectoral flap donor site. All 9 patients were successfully decannulated without shortness of breath, stridor, or recurrent stenosis at the time of follow-up. Allograft tracheoplasty is a new technique for the reconstruction of recurrent tracheal stenosis. It appears to be reliable for extensive airway defects that are refractory to conventional tracheoplasty techniques.


Enantioselective Synthesis of Indole-Annulated Medium-Sized Rings.
Asymmetric synthesis of indole-annulated medium-sized-ring compounds is developed through an iridium-catalyzed allylic dearomatization/retro-Mannich/hydrolysis cascade reaction. The reaction features mild conditions and a broad substrate scope. Under the optimal conditions, various seven-, eight-, or nine-membered-ring compounds can be afforded in good to excellent yields and excellent enantioselectivity. The proposed mechanism is supported by capturing the dearomatized intermediate through in situ reduction.


Top-down and bottom-up mechanisms in biasing competition in the human brain.
The biased competition theory of selective attention has been an influential neural theory of attention, motivating numerous animal and human studies of visual attention and visual representation. There is now neural evidence in favor of all three of its most basic principles: that representation in the visual system is competitive; that both top-down and bottom-up biasing mechanisms influence the ongoing competition; and that competition is integrated across brain systems. We review the evidence in favor of these three principles, and in particular, findings related to six more specific neural predictions derived from these original principles.


[Plasmodium falciparum drug resistance in the Congo. Evaluation of surveys carried out from 1985 to 1989].
Surveys on drug sensitivity of Plasmodium falciparum carried out between 1985 and 1989 included 7-day in vitro tests and in vivo tests. 485 in vivo tests were carried out in eight surveys conducted in Brazzaville and in several inland regions. The subjects were congolese children aged between 3 months and 15 years old. They were recruited in hospital, mother-child clinics or at school. The drugs studied were chloroquine, amodiaquine and the sulfadoxine-pyrimethamine combination. 182 strains were tested in vitro in two surveys (December 1985 and January 1987); amino-4-quinolines, quinine and mefloquine were studied. Although resistance to amino-4-quinolines is a recent occurrence, by 1985 it had spread widely in the indigenous population in the Centre and South of the country. Resistance has since increased gradually, especially for chloroquine which undergoes specific surveillance. The situation is less serious in the North, a less densely populated region which is still enclosed. In an in vivo comparative study with chloroquine conducted in Brazzaville in November 1986, amodiaquine was found to be only slightly more effective at a similar dosage. At that time, certain isolated observations already seem to imply that the sulfadoxine-pyrimethamine combination was also affected by resistance. This was not corroborated in an in vivo study carried out in 1989 on 40 children presenting with a malarial attack. Although the sensitivity to quinine may probably be decreased. This drug cannot yet be considered as being truly affected by resistance. The activity of mefloquine, the use of which is still limited, was satisfactory in 1987 in two different regions of the country.


Impact of deep coalescence on the reliability of species tree inference from different types of DNA markers in mammals.
An important challenge for phylogenetic studies of closely related species is the existence of deep coalescence and gene tree heterogeneity. However, their effects can vary between species and they are often neglected in phylogenetic analyses. In addition, a practical problem in the reconstruction of shallow phylogenies is to determine the most efficient set of DNA markers for a reliable estimation. To address these questions, we conducted a multilocus simulation study using empirical values of nucleotide diversity and substitution rates obtained from a wide range of mammals and evaluated the performance of both gene tree and species tree approaches to recover the known speciation times and topological relationships. We first show that deep coalescence can be a serious problem, more than usually assumed, for the estimation of speciation times in mammals using traditional gene trees. Furthermore, we tested the performance of different sets of DNA markers in the determination of species trees using a coalescent approach. Although the best estimates of speciation times were obtained, as expected, with the use of an increasing number of nuclear loci, our results show that similar estimations can be obtained with a much lower number of genes and the incorporation of a mitochondrial marker, with its high information content. Thus, the use of the combined information of both nuclear and mitochondrial markers in a species tree framework is the most efficient option to estimate recent speciation times and, consequently, the underlying species tree.


Nano-engineered living bacterial motors for active microfluidic mixing.
Active micromixers with rotating elements are attractive microfluidic actuators in many applications because of their mixing ability at a short distance. However, miniaturising the impeller design poses technical challenges including the fabrication and driving means. As a possible solution inspired by macro magnetic bar-stirrers, this study proposes the use of tethered, rotating bacteria as mixing elements. A tethered cell is a genetically engineered, harmless Escherichia coli (E. coli) attached to a surface by a single, shortened flagellum. The tethered flagellum acts as a pivot around which the entire cell body smoothly rotates. Videomicroscopy, image analysis and computational fluid dynamics (CFD) are utilised to demonstrate a proof-of-concept for the micro mixing process. Flow visualisation experiments show that a approximately 3 microm long tethered E. coli rotating at approximately 240 rpm can circulate a 1 microm polystyrene bead in the adjacent area at an average speed of nearly 4 microm/s. The Peclet (Peb) number for the stirred bead is evaluated to approximately 4. CFD simulations show that the rotary motion of a tethered E. coli rotating at 240 rpm can generate fluid velocities, up to 37 microm/s bordering the cell envelop. Based on these simulations, the Strouhal number (St) is calculated to about 2. This hybrid bio-inorganic micromxer could be used as a local, disposable mixer.


[Spatial Prediction Method of Farmland Soil Organic Matter in Weibei Dryland of Shaanxi Province].
Accurately predicting the spatial distribution of soil organic matter (SOM) content is of great significance for improving soil quality and improving the level of regional soil management. In order to explore the optimal model for predicting the SOM content of farmland in the Weibei Dryland of Shaanxi Province, the influence factors closely related to SOM content were selected as the modeling covariables, and a geographic detector, the ordinary kriging method (OK), geographic weighted regression model (GWR), partial least squares regression model (PLS), geographically weighted regression extended model (GWRPLS), and random forest model (RF) were used to predict the spatial distribution of SOM content in training samples. Additionally, the validation set samples were used to compare and analyze the prediction accuracy of the five methods. The results showed:① the main factors affecting the spatial variability of soil SOM were total nitrogen, fertilizer application, available potassium, available phosphorus, and altitude, and the interaction between any two factors was more explanatory for SOM than any single factor. ②ω(SOM) in farmland was between 2.25 and 30.23 g·kg-1, with an average value of 15.14 g·kg-1 and a coefficient of variation of 30.00. Although there were local differences in the prediction results of SOM by the five methods, the overall spatial distribution trend was basically the same. In the study area, the content of organic matter was low in the north and northeast and high in the west and southeast. ③ From the perspective of the prediction accuracy of the five methods, the root mean square error (RMSE) and mean absolute error (MAE) of RF were the smallest, and the prediction deviation (RPD) of GWRPLS was the largest. Compared with the OK method, the correlation coefficients (r) of GWR, PLS, RF, and GWRPLS increased to 0.907, 0.836, 0.968, and 0.972, respectively. Comprehensive analysis results showed that the random forest model had the highest prediction accuracy.


Assays for IGF binding proteins.
Growing interest in IGF binding proteins and their roles in the modulation of IGF actions has prompted several laboratories to establish immunological assays to measure these proteins. Three distinct IGF binding proteins, referred as IGFBP-1, IGFBP-2 and IGFBP-3, have so far had their cDNAs cloned and sequenced. The availability of purified proteins and the development of polyclonal and monoclonal antibodies now allow specific quantitation of different IGFBP classes. This brief review will address on the current progress in the immunological assays of IGFBPs. The importance of the sensitivity, the reproducibility, and the standardization of the assays is discussed. Some additional information, like levels in amniotic fluid and serum of healthy adults and children, is given.


The association between radiographic embrasure morphology and interdental papilla reconstruction using injectable hyaluronic acid gel.
The purpose of this study was to evaluate the clinical efficacy of enhancing deficient interdental papilla with hyaluronic acid gel injection by assessing the radiographic anatomical factors affecting the reconstruction of the interdental papilla. Fifty-seven treated sites from 13 patients (6 males and 7 females) were included. Patients had papillary deficiency in the upper anterior area. Prior to treatment, photographic and periapical radiographic standardization devices were designed for each patient. A 30-gauge needle was used with an injection-assistance device to inject a hyaluronic acid gel to the involved papilla. This treatment was repeated up to 5 times every 3 weeks. Patients were followed up for 6 months after the initial gel application. Clinical photographic measurements of the black triangle area (BTA), height (BTH), and width (BTW) and periapical radiographic measurements of the contact point and the bone crest (CP-BC) and the interproximal distance between roots (IDR) were undertaken using computer software. The interdental papilla reconstruction rate (IPRR) was calculated to determine the percentage change of BTA between the initial and final examination and the association between radiographic factors and the reconstruction of the interdental papilla by means of injectable hyaluronic acid gel were evaluated. All sites showed improvement between treatment examinations. Thirty-six sites had complete interdental papilla reconstruction and 21 sites showed improvement ranging from 19% to 96%. The CP-BC correlated with the IPRR. More specifically, when the CP-BC reached 6 mm, virtually complete interdental papilla reconstruction via injectable hyaluronic acid gel was achieved. These results suggest that the CP-BC is closely related to the efficacy of hyaluronic acid gel injection for interdental papilla reconstruction.


Comparative landscape genetics reveals the evolution of viviparity reduces genetic connectivity in fire salamanders.
Evolutionary changes in reproductive mode may affect co-evolving traits, such as dispersal, although this subject remains largely underexplored. The shift from aquatic oviparous or larviparous reproduction to terrestrial viviparous reproduction in some amphibians entails skipping the aquatic larval stage and, thus, greater independence from water. Accordingly, amphibians exhibiting terrestrial viviparous reproduction may potentially disperse across a wider variety of suboptimal habitats and increase population connectivity in fragmented landscapes compared to aquatic-breeding species. We investigated this hypothesis in the fire salamander (Salamandra salamandra), which exhibits both aquatic- (larviparity) and terrestrial-breeding (viviparity) strategies. We genotyped 426 larviparous and 360 viviparous adult salamanders for 13 microsatellite loci and sequenced a mitochondrial marker for 133 larviparous and 119 viviparous individuals to compare population connectivity and landscape resistance to gene flow within a landscape genetics framework. Contrary to our predictions, viviparous populations exhibited greater differentiation and reduced genetic connectivity compared to larviparous populations. Landscape genetic analyses indicate viviparity may be partially responsible for this pattern, as water courses comprised a significant barrier only in viviparous salamanders, probably due to their fully terrestrial life cycle. Agricultural areas and, to a lesser extent, topography also decreased genetic connectivity in both larviparous and viviparous populations. This study is one of very few to explicitly demonstrate the evolution of a derived reproductive mode affects patterns of genetic connectivity. Our findings open avenues for future research to better understand the eco-evolutionary implications underlying the emergence of terrestrial reproduction in amphibians.


In Silico Identification of Contradictory Role of ADAMTS5 in Hepatocellular Carcinoma.
ADAMTS5 has different roles in multiple types of cancers and participates in various molecular mechanisms. However, the prognostic value of ADAMTS5 in patients with hepatocellular carcinoma (HCC) still remains unclear. We carried the study to evaluate the prognostic value and identified underlying molecular mechanisms in HCC. Firstly, the association of ADAMTS5 expression and clinicopathological parameters was evaluated by in GSE14520. Next, ADAMTS5 expression in HCC was performed using GSE14520, GSE36376, GSE76427 and The Cancer Genome Atlas (TCGA) profile. Furthermore, Kaplan-Meier analysis, Univariate and Multivariate Cox regression analysis, subgroup analysis was performed to evaluate the prognostic value of ADAMTS5 in HCC. Finally, GO enrichment analysis, gene set enrichment analysis (GSEA) and weighted gene co-expression network analysis (WGCNA) were performed to revealed underlying molecular mechanisms. The expression of ADAMTS5 was positively correlated with the development of HCC. Next, high ADAMTS5 expression was significantly associated with poorer survival (all P < 0.05) and the impact of ADAMTS5 on all overall survival (OS), disease-free survival (DFS), relapse-free survival (RFS), disease specific survival (DSS) and progression free interval (PFI) was specific for HCC among other 29 cancer types. Subgroup analysis showed that ADAMTS5 overexpression was significantly associated with poorer OS in patients with HCC. Finally, ADAMTS5 might participate in the status conversion from metabolic-dominant to extracellular matrix-dominant, and the activation of ECM-related biological process might contribute to high higher mortality risk for patients with HCC. ADAMTS5 may play an important role in the progression of HCC, and may be considered as a novel and effective biomarker for predicting prognosis for patients with HCC.


[Study on the prediction of soil heavy metal elements content based on mid-infrared diffuse reflectance spectra].
The present paper analyzed the possibility of mid-infrared diffuse reflectance spectra for quick assessment of heavy metal element content in soil quickly. Soil samples were collected from Jiangning District and Baguazhou Island, and the numbers of sample were 103 and 58 separately. Jiangning District samples were used as calibration set while Baguazhou Island samples as validation set. To assess the utility of different pre-treatment process of MIR spectroscopy for soil heavy metal element content analysis, we used PLSR method to develop the calibration between spectral data and soil elements content. Three spectral pretreating techniques such as smooth, log(1/N), baseline correction, multiplicative scatter correction were used for promotion of predicting performance. The result showed that the progress of (log-BC-MSC) in turn achieved optimal calibration of MIR spectra and better prediction for ex-situ soils. Though the calibration data were treated by different pre-treating schema, the R2 of the 8 elements followed the same law: Ni > 0.8 > Cr, Cu, Zn, Pb, Hg > 0.6 > As, Cd. When we applied these calibrations to Baguazhou Island soils, (log-BC-MSC) treated data results in the smallest RMSEp-BGZ. We used the same calibration method to compare the predictive ability of MIR spectra to VNIR spectra. The R2 of 8 elements developed by VNIR spectral calibration are sometime larger than MIR's, but after we applied these calibrations to validation set, the RSME of MIR data for prediction of BGZ soil samples is 21% to 73% of VNIR's. This result showed us that for predicting ex-situ soils, MIR analysis substantially outperformed VNIR These results indicated that MIR spectra can be used to predict soil heavy metal content quickly and non-destructively.


Stent Thrombosis.
The most dreaded complication with percutaneous coronary intervention with stents, either bare-metal or drug-eluting stents is stent thrombosis (ST) and it has a significant detrimental effect on the outcome for the patient. The initial attempts at intervention with bare-metal stents had much higher rates of ST compared with what is currently prevailing in the modern interventional world. Significant changes with respect to the stent technology, pharmacology, and most importantly our understanding of this phenomenon have decreased the risk of ST. There are many factors that can be performed to minimize the risk of ST and this review will describe the incidence, pathophysiology, and contributing risk factors to ST.


Hematopoietic stem cell gene therapy for inherited bone marrow disorders: past accomplishments and continued challenges.
From the time that the genes encoding the defective proteins were cloned for a number of inherited diseases, it became a goal to correct those conditions by restoring the normal gene and thereby, its product. For the inherited disorders affecting the blood and its progenitor cells, the hematopoietic stem cells were the ideal target cells for gene transfer, because the normal gene would then be transferred to all of the progeny cells, theoretically for the lifetime of the recipient. However, the tasks of isolating the hematopoietic stem cells, introducing the new genes in such a manner as to preserve engraftment of the manipulated cells, and achieving long-term gene expression, have not been straightforward in the clinical trial setting, although there has been moderate success for cells in vitro, and in murine studies. With the report of clinical efficacy of gene transfer in children with X-linked severe combined immunodeficiency disease, the dream of clinical gene transfer to hematopoietic cells has become a reality. But there are still significant impediments remaining for a number of diseases. The innovations of introduction of synthetic receptors that confer growth advantage, the use of lentiviral vectors with increased stem cell transduction efficiency, and the addition of modified promoter/enhancer sequences to augment and preserve gene expression may bring wider success to gene therapy clinical trials for bone marrow disorders in the near future.


Functional exploration of the adult ovarian granulosa cell tumor-associated somatic FOXL2 mutation p.Cys134Trp (c.402C>G).
The somatic mutation in the FOXL2 gene c.402C>G (p.Cys134Trp) has recently been identified in the vast majority of adult ovarian granulosa cell tumors (OGCTs) studied. In addition, this mutation seems to be specific to adult OGCTs and is likely to be a driver of malignant transformation. However, its pathogenic mechanisms remain elusive. We have sequenced the FOXL2 open reading frame in a panel of tumor cell lines (NCI-60, colorectal carcinoma cell lines, JEG-3, and KGN cells). We found the FOXL2 c.402C>G mutation in the adult OGCT-derived KGN cell line. All other cell lines analyzed were negative for the mutation. In order to gain insights into the pathogenic mechanism of the p.Cys134Trp mutation, the subcellular localization and mobility of the mutant protein were studied and found to be no different from those of the wild type (WT). Furthermore, its transactivation ability was in most cases similar to that of the WT protein, including in conditions of oxidative stress. A notable exception was an artificial promoter known to be coregulated by FOXL2 and Smad3, suggesting a potential modification of their interaction. We generated a 3D structural model of the p.Cys134Trp variant and our analysis suggests that homodimer formation might also be disturbed by the mutation. Here, we confirm the specificity of the FOXL2 c.402C>G mutation in adult OGCTs and begin the exploration of its molecular significance. This is the first study demonstrating that the p.Cys134Trp mutant does not have a strong impact on FOXL2 localization, solubility, and transactivation abilities on a panel of proven target promoters, behaving neither as a dominant-negative nor as a loss-of-function mutation. Further studies are required to understand the specific molecular effects of this outstanding FOXL2 mutation.


Structural insight into the mechanisms of targeting and signaling of focal adhesion kinase.
Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase whose focal adhesion targeting (FAT) domain interacts with other focal adhesion molecules in integrin-mediated signaling. Localization of activated FAK to focal adhesions is indispensable for its function. Here we describe a solution structure of the FAT domain bound to a peptide derived from paxillin, a FAK-binding partner. The FAT domain is composed of four helices that form a "right-turn" elongated bundle; the globular fold is mainly maintained by hydrophobic interactions. The bound peptide further stabilizes the structure. Certain signaling events such as phosphorylation and molecule interplay may induce opening of the helix bundle. Such conformational change is proposed to precede departure of FAK from focal adhesions, which starts focal adhesion turnover.


Boron nitride nanotubes and primary human osteoblasts: in vitro compatibility and biological interactions under low frequency ultrasound stimulation.
In this paper we investigated a novel and non-invasive approach for an endogenous osteoblast stimulation mediated by boron nitride nanotubes (BNNTs). Specifically, following the cellular uptake of the piezoelectric nanotubes, cultures of primary human osteoblasts (hOBs) were irradiated with low frequency ultrasound (US), as a simple method to apply a mechanical input to the cells loaded with BNNTs. This in vitro study was aimed at investigating the main interactions between hOBs and BNNTs and to study the effects of the 'BNNTs + US' stimulatory method on the osteoblastic function and maturation.A non-cytotoxic BNNT concentration to be used in vitro with hOB cultures was established. Moreover, investigation with transmission electron microscopy/electron energy loss spectroscopy (TEM/EELS) confirmed that BNNTs were internalized in membranal vesicles. The panel of investigated osteoblastic markers disclosed that BNNTs were capable of fostering the expression of late-stage bone proteins in vitro, without using any mineralizing culture supplements. In our samples, the maximal osteopontin expression, with the highest osteocalcin and Ca(2+) production, in the presence of mineral matrix with nodular morphology, was observed in the samples treated with BNNTs + US. In this group was also shown a significantly enhanced synthesis of TGF-β1, a molecule sensitive to electric stimulation in bone. Finally, gene deregulations of the analyzed osteoblastic genes leading to depletive cellular effects were not detected. Due to their piezoelectricity, BNNT-based therapies might disclose advancements in the treatment of bone diseases.


Pertussis outbreak in Papua New Guinea: the challenges of response in a remote geo-topographical setting.
A large outbreak of pertussis was detected during March 2011 in Goilala, a remote district of the Central Province in Papua New Guinea, characterized by rugged topography with no road access from the provincial headquarters. This outbreak investigation highlights the difficulties in reporting and responding to outbreaks in these settings. The suspected pertussis cases, reported by health workers from the Ononge health centre area, were investigated and confirmed for the presence of Bordetella pertussis DNA using the polymerase chain reaction (PCR) method. There were 205 suspected pertussis cases, with a case-fatality rate (CFR) of 3%. All cases were unvaccinated. The Central Province conducted a response vaccination programme providing 65% of children less than five years of age with diphtheria-pertussis-tetanus-HepB-Hib vaccine at a cost of US$ 12.62 per child. The incurred cost of vaccination in response to this outbreak was much higher than the US$ 3.80 per child for routine outreach patrol. To prevent further outbreaks of vaccine-preventable diseases in these areas, local health centres must ensure routine vaccination is strengthened through the "Reaching Every District" initiative of the National Department of Health.


Nitrendipine once or twice daily in hypertensive non-insulin-dependent diabetes mellitus.
Twenty (12 male, 8 female) hypertensive patients were recruited into the study; their mean (+/- SD) sitting blood pressure (BP) was 166 +/- 13/101 +/- 4 mm Hg, age 59.6 +/- 5.8 years, weight 83.4 +/- 14.2 kg, and glycosylated hemoglobin (HbA1) 9.9 +/- 2.6%. After a run-in period of 4 weeks, patients were randomized to receive nitrendipine once or twice daily, with 15 patients completing the 12-week study period. Prior to treatment with nitrendipine, the patients subsequently treated on a twice-daily regimen had a higher sitting systolic BP (SBP) of 173 +/- 15 mm Hg compared to the once-daily patient group at 159 +/- 7 mm Hg (p less than 0.05). There were no differences between the two treatment groups in weight or diabetes control. Once-daily nitrendipine effectively reduced the sitting SBP and diastolic BP (DBP), i.e., SBP fell from 159 +/- 7 to 147 +/- 12 mm Hg (p less than 0.05) and DBP fell from 100 +/- 4 to 86.3 +/- 8 mm Hg (p less than 0.001). Nonsignificant reductions in weight from 81.5 +/- 11.3 to 80.9 +/- 11.2 kg and HbA1 from 9.8 +/- 3.3 to 8.4 +/- 1.3% were also seen. Similarly, with twice-daily nitrendipine, a significant (p less than 0.01) fall in sitting systolic BP from 173 +/- 15 to 148 +/- 9 mm Hg and DBP 103 +/- 4 to 85 +/- 8 mm Hg was also achieved. Little or no change was observed in weight or glycemic control in this patient group.(ABSTRACT TRUNCATED AT 250 WORDS)


Detection of nonstructural protein 6 in murine coronavirus-infected cells and analysis of the transmembrane topology by using bioinformatics and molecular approaches.
Coronaviruses encode large replicase polyproteins which are proteolytically processed by viral proteases to generate mature nonstructural proteins (nsps) that form the viral replication complex. Mouse hepatitis virus (MHV) replicase products nsp3, nsp4, and nsp6 are predicted to act as membrane anchors during assembly of the viral replication complexes. We report the first antibody-mediated Western blot detection of nsp6 from MHV-infected cells. The nsp6-specific peptide antiserum detected the replicase intermediate p150 (nsp4 to nsp11) and two nsp6 products of approximately 23 and 25 kDa. Analysis of nsp6 transmembrane topology revealed six membrane-spanning segments and a conserved hydrophobic domain in the C-terminal cytosolic tail.


Cytoskeletal Mechanics Regulating Amoeboid Cell Locomotion.
Migrating cells exert traction forces when moving. Amoeboid cell migration is a common type of cell migration that appears in many physiological and pathological processes and is performed by a wide variety of cell types. Understanding the coupling of the biochemistry and mechanics underlying the process of migration has the potential to guide the development of pharmacological treatment or genetic manipulations to treat a wide range of diseases. The measurement of the spatiotemporal evolution of the traction forces that produce the movement is an important aspect for the characterization of the locomotion mechanics. There are several methods to calculate the traction forces exerted by the cells. Currently the most commonly used ones are traction force microscopy methods based on the measurement of the deformation induced by the cells on elastic substrate on which they are moving. Amoeboid cells migrate by implementing a motility cycle based on the sequential repetition of four phases. In this paper we review the role that specific cytoskeletal components play in the regulation of the cell migration mechanics. We investigate the role of specific cytoskeletal components regarding the ability of the cells to perform the motility cycle effectively and the generation of traction forces. The actin nucleation in the leading edge of the cell, carried by the ARP2/3 complex activated through the SCAR/WAVE complex, has shown to be fundamental to the execution of the cyclic movement and to the generation of the traction forces. The protein PIR121, a member of the SCAR/WAVE complex, is essential to the proper regulation of the periodic movement and the protein SCAR, also included in the SCAR/WAVE complex, is necessary for the generation of the traction forces during migration. The protein Myosin II, an important F-actin cross-linker and motor protein, is essential to cytoskeletal contractility and to the generation and proper organization of the traction forces during migration.


[Amplification of L-myc oncogene in malignant meningioma. Case report].
Amplification of L-myc oncogene was noticed in a malignant meningioma originating from the right sphenoidal wing of a 54-year-old female. The patient underwent three surgical resections plus radiotherapy over a period of 11 years and then the growth rate of the tumor became much greater with a severely invasive appearance. Using Southern blot hybridization, L-myc amplification was examined on the specimen resected at the fourth operation. As a result, approximately five-fold amplification was confirmed, which has not been previously reported except for that in a small cell carcinoma of the lung. This result may suggest that L-myc amplification is responsible to some extent for the malignant transformation in this meningioma.


Banknote authenticity is signalled by rapid neural responses.
Authenticating valuable objects is widely assumed to involve protracted scrutiny for detection of reproduction flaws. Yet, accurate authentication of banknotes is possible within one second of viewing, suggesting that rapid neural processes may underpin counterfeit detection. To investigate, we measured event-related brain potentials (ERPs) in response to briefly viewed genuine or forensically recovered counterfeit banknotes presented in a visual oddball counterfeit detection task. Three ERP components, P1, P3, and extended P3, were assessed for each combination of banknote type (genuine, counterfeit) and overt response ("real", "fake"). P1 amplitude was greater for oddballs, demonstrating that the initial feedforward sweep of visual processing yields the essential information for differentiating genuine from counterfeit. A similar oddball effect was found for P3. The magnitude of this P3 effect was positively correlated with behavioural counterfeit sensitivity, although the corresponding correlation for P1 was not. For the extended P3, amplitude was greatest for correctly detected counterfeits and similarly small for missed counterfeits, incorrectly and correctly categorised genuine banknotes. These results show that authentication of complex stimuli involves a cascade of neural processes that unfolds in under a second, beginning with a very rapid sensory analysis, followed by a later decision stage requiring higher level processing.


Epidemiology of the metabolic syndrome in Hungary.
Hungary has high cardiovascular mortality. Recent studies have revealed a high prevalence of several cardiovascular risk factors, including obesity, diabetes and hypertension. The objective of this study was to assess the prevalence of the metabolic syndrome in Hungary. Cross-sectional study. Within the framework of the Hungarian General Practitioners' Morbidity Sentinel Stations Programme, a random sample of 2006 individuals aged 20-69 years was selected in 2006. Physical examinations, blood sampling and data collection were performed by general practitioners. Information on environmental factors was gathered using a questionnaire. The population prevalence was estimated based on the sample frequencies. The overall response rate was 91%. The age-adjusted prevalence of the metabolic syndrome using the 2009 Harmonized definition was 38% [95% confidence interval (CI) 35-42%] in males and 30% (95% CI 28-33%) in females aged 20-69 years. There were no significant regional differences in the frequency figures. The high prevalence of the metabolic syndrome is a serious public health problem in Hungary, and remains a major determinant of the high burden of cardiovascular disease.


The chloroplast genome sequence of mungbean (Vigna radiata) determined by high-throughput pyrosequencing: structural organization and phylogenetic relationships.
Mungbean is an economically important crop which is grown principally for its protein-rich dry seeds. However, genomic research of mungbean has lagged behind other species in the Fabaceae family. Here, we reported the complete chloroplast (cp) genome sequence of mungbean obtained by the 454 pyrosequencing technology. The mungbean cp genome is 151 271 bp in length which includes a pair of inverted repeats (IRs) of 26 474 bp separated by a small single-copy region of 17 427 bp and a large single-copy region of 80 896 bp. The genome contains 108 unique genes and 19 of these genes are duplicated in the IR. Of these, 75 are predicted protein-coding genes, 4 ribosomal RNA genes and 29 tRNA genes. Relative to other plant cp genomes, we observed two distinct rearrangements: a 50-kb inversion between accD/rps16 and rbcL/trnK-UUU, and a 78-kb rearrangement between trnH/rpl14 and rps19/rps8. We detected sequence length polymorphism in the cp homopolymeric regions at the intra- and inter-specific levels in the Vigna species. Phylogenetic analysis demonstrated a close relationship between Vigna and Phaseolus in the phaseolinae subtribe and provided a strong support for a monophyletic group of the eurosid I.


Sexual Orientation Disparities in Risk Factors for Adverse COVID-19-Related Outcomes, by Race/Ethnicity - Behavioral Risk Factor Surveillance System, United States, 2017-2019.
Sexual minority persons experience health disparities associated with sexual stigma and discrimination and have a high prevalence of several health conditions that have been associated with severe coronavirus disease 2019 (COVID-19) (1,2). Current COVID-19 surveillance systems do not capture information about sexual orientation. To begin bridging the gap in knowledge about COVID-19 risk among sexual minority adults, CDC examined disparities between sexual minority and heterosexual adults in the prevalence of underlying conditions with strong or mixed evidence of associations with severe COVID-19-related illness (3), by using data from the 2017-2019 Behavioral Risk Factor Surveillance System (BRFSS).* When age, sex, and survey year are adjusted, sexual minority persons have higher prevalences than do heterosexual persons of self-reported cancer, kidney disease, chronic obstructive pulmonary disease (COPD), heart disease (including myocardial infarction, angina, or coronary heart disease), obesity, smoking, diabetes, asthma, hypertension, and stroke. Sexual minority adults who are members of racial/ethnic minority groups disproportionately affected by the pandemic also have higher prevalences of several of these health conditions than do racial/ethnic minority adults who are heterosexual. Collecting data on sexual orientation in COVID-19 surveillance and other studies would improve knowledge about disparities in infection and adverse outcomes by sexual orientation, thereby informing more equitable responses to the pandemic.


Four new species of Cymadusa Savigny, 1816 (Amphipoda: Ampithoidae) and new records of C. filosa Savigny, 1816 from Brazilian coast.
Four new species of the genus Cymadusa are described from Brazil (states of Ceará, Espírito Santo and Rio de Janeiro). Cymadusa icapui is described from Ceará state, C. trindadensis is described from Espírito Santo state, C. rasae and C. tartarugae are described from Rio de Janeiro state. Two identification keys are presented: one to all known species of the family Ampithoidae from Brazil, and another one to world species of Cymadusa. Based on literary review and analysis of collection material, we extend the distribution of Cymadusa filosa Savigny, 1816 along the Brazilian coast by finding new location records of the species based on analysis of stored collection material.


Clinical manifestation and current therapeutics in X-juvenile retinoschisis.
X-linked juvenile retinoschisis (XLRS) is one of the common early-onset hereditary retinal degenerative diseases in men. The common symptoms of XLRS range from mild to severe central vision loss and radial stripes created by the fovea, the division of the inner layer of the retina in the peripheral retina and the significant decrease in b-wave amplitude (ERG). Retinoschisin, the 224-amino-acid protein product of the retinoschisis 1 (RS1) gene, contains a discoid domain as the primary structural unit, an N-terminal cleavable signal sequence, and an oligomerization-area component. Retinoschisin is a homo-octamer complex with disulfide links that are released by retinal cells. It helps preserve the retina's integrity by binding to the surface of photoreceptors and bipolar cells. As a recessive genetic disease, XLRS was usually treated by prescribing low vision aids in most clinical cases. A gene replacement therapy based on adeno-associated virus vectors was initiated and showed a breakthrough in treating XLRS in 2014. Understanding the revolution of gene therapy for treating XLRS may accelerate its development and make this gene therapy the template for developing therapeutics against other inherited retinal diseases.


Diagnostic value of the combined detection of CEA, NSE and IL-18 for lung cancer and their relationship with apoptosis gene Bcl-2.
This study aims to investigate the value of the combined detection of carcinoembryonic antigen (CEA), Neuron-specific enolase (NSE) and the level of Interleukin-18 (IL-18) in the serum in the diagnosis of lung cancer. The correlation between these parameters and the expression levels of B-cell lymphoma-2 (Bcl-2) protein were also studied. Eighty patients with lung cancer were included in the lung cancer group. These patients underwent surgery in the Department of Oncology of Huai'an Second People's Hospital between February 2016 and February 2018. During the same period, another 80 patients with benign lung lesions were registered in the benign lesion group and 80 healthy people were enrolled in the control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of CEA, NSE and IL-18. The diagnostic critical value of these factors was used as positive indicator. When CEA, NSE and IL-18 levels were positive at the same time, the combined detection was considered to be positive. WB was used to detect Bcl-2 expression level. We also analyzed the possible correlation between CEA, NSE, IL-18 levels and the Bcl-2 expression levels. The CEA, NSE and IL-18 expression levels in the serum of the lung cancer group were significantly higher than those in the benign lesion and the control groups (p<0.05). The area under ROC curve for CEA, NSE and IL-18 respectively was 0.770 (0.697-0.843), 0.829 (0.767-0.890), 0.721 (0.642-0.800) (p<0.001). IL-18 level was negatively correlated with the level of Bcl-2 mRNA in the tissue (r=-0.380, p<0.001). In conclusion, CEA, NSE and IL-18 have a good auxiliary diagnostic value in patients with lung cancer. The combined detection could improve the sensitivity and specificity of lung can¬cer diagnosis. There was a negative correlation between IL-18 and Bcl-2 levels which suggested a potential inhibitory role of IL-18 on the lung cancer cells apoptosis pathway.


The COVID-19 Pandemic and the TRIPS Waiver: Patents and Flexibility.
The World Trade Organization's Agreement on Trade-Related Aspects of Intellectual Property Rights (TRIPS) provides for global minimum standard patents. These patents potentially limit access to products and processes for the surveillance, tracking, diagnosis and treatment of COVID-19. A possible solution currently under consideration is a TRIPS waiver of the implementation, application and enforcement for the prevention, containment or treatment of COVID-19. This article addresses the ways that TRIPS patents might be mediated including through TRIPS flexibilities. The article argues that there are sufficient means of derogating from patents (and potentially copyright, industrial designs and undisclosed information), although they alone will not resolve the access problems. The article concludes that the key patent problem is the transfer of know-how and that developing new ideas about addressing these patent know-how transfers is the presently unaddressed challenge.


Sunitinib in the Treatment of Thyroid Cancer.
Sunitinib (SU11248) is an oral multi-target tyrosine kinase inhibitor (TKI) with low molecular weight, that inhibits platelet-derived growth factor receptors (PDGF-Rs) and vascular endothelial growth factor receptors (VEGFRs), c-KIT, fms-related tyrosine kinase 3 (FLT3) and RET. The concurrent inhibition of these pathways reduces tumor vascularization and causes cancer cell apoptosis, inducing a tumor shrinkage. Sunitinib is approved for the treatment of imatinib-resistant gastrointestinal stromal tumor (GIST), renal carcinoma, and pancreatic neuroendocrine tumors. We searched the literature on PubMed library. In vitro studies showed that sunitinib targeted the cytosolic MEK/ERK and SAPK/JNK pathways in the RET/PTC1 cell inhibiting cell proliferation and causing stimulation of sodium/iodide symporter (NIS) gene expression in RET/PTC1 cells. Furthermore sunitinib is active in vitro and in vivo against anaplastic thyroid cancer (ATC) cells. Most of the clinical studies report that sunitinib is effective as first- and second-line TKI therapy in patients with advanced dedifferentiated thyroid cancer (DeTC), or medullary thyroid cancer (MTC). Sunitinib 37.5 mg/day is well tolerated, and effective. The most common adverse events include: reduction in blood cell counts (in particular leukocytes), hand-foot skin reaction, diarrhea, fatigue, nausea, hypertension, and musculoskeletal pain. Even if sunitinib is promising in the therapy of differentiated thyroid carcinoma (DTC), until now no phase III studies have been published, and additional prospective researches are necessary in order to evaluate the real efficacy of sunitinib in aggressive thyroid cancer.


[Airway obstruction caused by large blood vessel anomalies: assessment by flexible bronchoscopy].
To evaluate the diagnostic value and safety of flexible bronchoscopy in congenital great vessel diseases complicated with airway compression. The medical records of patients with great vessels abnormalities who were admitted to the neonatal intensive care unit (NICU) from October 2005 to June 2009 were retrospectively reviewed; 34 cases were diagnosed as airway compression by flexible bronchoscopy, 10 cases as vascular ring, 24 cases as aortal arch obstruction. The age of the patients was 6 d - 11 m, body weight 2.2 - 8.7 kg [(4.6 +/- 1.4) kg]. Recorded airway abnormalities detected by bronchoscopy and CT, cardiac vascular defects and airway compression were consistent with the findings on operation. The relation between the airway compression and cardiac vascular abnormalities, treatment of the airway compression and outcome were analysed. Bronchoscopic assessment was successfully performed in NICU or operating room for all the patients. (1) Initial presentation of the 34 cases were tachypnea, stridor, refractory lung infection and prolonged mechanical ventilation. (2) Extrinsic compression was found in all the 10 cases with vascular ring by bronchoscopy initially which indicated vascular ring, airway compression was mainly of lower part of trachea. Diagnosis of 9 cases was consistent with CT diagnosis and in 1 case the diagnosis was confirmed by surgery; among these cases, 7 had congenital tracheal stenosis. (3) In the 24 cases with aortic obstructive lesion, 5 were detected to have tracheal stenosis by CT before correction of vascular abnormality, among whom one case was indicated to have tracheal stenosis by bronchoscopy, the other 19 cases were found with airway compression by bronchoscopy during or after vascular correction. Among the 24 cases, 21 had left main bronchial stenosis, 2 had congenital tracheal stenosis. Airway compression diagnosed by bronchoscopy agreed with the findings of CT. Two cases developed transient decrease of oxygen saturation, 5 cases developed transient tachycardia. Flexible bronchoscopy plays an important role in assessment of the airway compression complicated with great vessel abnormalities. Bronchoscopy is an accurate, convenient, safe and rapid way for airway assessment, but further examination of the peripheral structure and vascular malformation need combined examination with CT.


Autochthonous and Travel Acquired Hepatitis E Virus in Australia.
Hepatitis E virus (HEV) is a common cause of acute viral hepatitis with significant morbidity and mortality, particularly in pregnant women. There are four major genotypes which can cause disease in humans. Genotypes 1 and 2 are usually associated with outbreaks and spread via facal/oral route or contaminated water. Genotypes 3 and 4 are zoonotic and usually associated with handling of pigs or consumption of contaminated pork. The strains circulating in Australia have never been characterized. The aims for this project are to identify the HEV genotypes found in Australia and link them to possible sources of transmission by phylogenetic analysis. Between 2015 and 2020, 91 HEV isolates were sequenced and genotyped using an in-house PCR. Sixty-six of these were also sequenced by using the international HEVnet primers. Genotypes were determined using the BLASTn program. Relatedness to other strains in Australia was determined by phylogenetic analyses of the HEVnet sequences. Isolates were also stratified by state of origin, gender, age, predisposing factors and travel history (if known). Of the 91 HEV isolates sequenced, 55 (60.4%) were genotype 1. There were 34 (37.4%) genotype 3 strains and two genotype 4 (2.2%). At least 20 of the genotype 1 strains have been linked to travel in India, and another three with Pakistan. Five of the "Indian" strains were closely related and are suspected to have originated in Gujarat. Phylogenetic analysis also showed that 12 genotype 3 strains were genetically related and potentially acquired in/from New South Wales, Australia. The two genotype 4 strains may have originated in China. This is the first study to describe the HEV isolates identified in Australia. The results infer that HEV may be acquired during overseas travel as well as locally, presumably from consumption of pork or pork-related products. The phylogenetic analyses also reveal clusters of infection originating from India and Pakistan. This study provides some insight into the source and epidemiology of HEV infection in Australia which may be used to guide public health procedure and enable the implementation of measures to deal with potential outbreaks of infection.


[Effects of transgenic Bt crops on non-target soil animals].
Transgenic Bt crops are widely planted around the world. With the quick development and extension of genetically modified crops, it is needed to make a deep study on the effects of Bt crops on soil ecosystem. This paper reviewed the research progress on the effects of transgenic Bt crops on the population dynamics and community structure of soil animals, e.g., earthworm, nematode, springtail, mite, and beetle, etc. The development history of Bt crops was introduced, the passway the Bt protein comes into soil as well as the residual and degradation of Bt protein in soil were analyzed, and the critical research fields about the ecological risk analysis of transgenic Bt crops on non-target soil animals in the future were approached, which would provide a reference for the research of the effects of transgenic Bt crops on non-target soil animals.


Targeting kinase signaling pathways with constrained peptide scaffolds.
Kinases are amongst the largest families in the human proteome and serve as critical mediators of a myriad of cell signaling pathways. Since altered kinase activity is implicated in a variety of pathological diseases, kinases have become a prominent class of proteins for targeted inhibition. Although numerous small molecule and antibody-based inhibitors have already received clinical approval, several challenges may still exist with these strategies including resistance, target selection, inhibitor potency and in vivo activity profiles. Constrained peptide inhibitors have emerged as an alternative strategy for kinase inhibition. Distinct from small molecule inhibitors, peptides can provide a large binding surface area that allows them to bind shallow protein surfaces rather than defined pockets within the target protein structure. By including chemical constraints within the peptide sequence, additional benefits can be bestowed onto the peptide scaffold such as improved target affinity and target selectivity, cell permeability and proteolytic resistance. In this review, we highlight examples of diverse chemistries that are being employed to constrain kinase-targeting peptide scaffolds and highlight their application to modulate kinase signaling as well as their potential clinical implications.


Immunohistochemical demonstration of neuronal and astrocytic markers and oncofoetal antigens in retinoblastomas.
General opinion is that retinoblastomas, though not everyone agrees with that view. Some authors suggest that retinoblastomas are derived from a primitive retinal cell able to differentiate into both neuronal and glial cell lines. The aim of the present work was to study immunohistochemically the expression of neuronal and astrocytic markers in retinoblastomas and at the same time the presence of the oncofoetal antigens carcinoembryonic antigen (CEA) and alpha Foeto Protein (AFP), since patients with retinoblastomas often show high oncofoetal antigen in serum levels. For this purpose we employed the streptavidin-biotin immunoperoxidase technique in 13 cases of retinoblastoma to evaluate the presence and distribution of neuron-specific enolase (NSE), neurofilament protein (NF), glial fibrillary acidic protein (GFAP), S-100 protein, CEA and AFP. All 13 tumours studied stained for NSE. Seven of them showed GFAP- and S-100 positive perivascular glial cells as well as cells distributed randomly in the tumour that were interpreted as non tumour cells. All 13 retinoblastomas lacked detectable NF, CEA, and AFP. These results support the idea that retinoblastomas are neuronal tumours, although retinal glial cells may become incorporated in the tumour and proliferate in response to the tumour.


A crossover interaction between sex, sexual orientation, and handedness.
The distribution of handedness scores was analysed in 1414 heterosexual, bisexual, and homosexual men and women. Hand preference was assessed by a questionnaire that distinguishes consistently right-handed from partially or completely left-handed individuals, and sexual orientation was measured by the Kinsey scales. As expected from population-based studies, heterosexual men were, on average, more left-handed than heterosexual women. By contrast, gay men were more right-handed than lesbians or heterosexual men, and lesbians were more left-handed than gay men or heterosexual women. This crossover interaction suggests that a common variable influences sex, sexual orientation, and hand preference.


Risk for recurrent coronary heart disease and all-cause mortality among individuals with chronic kidney disease compared with diabetes mellitus, metabolic syndrome, and cigarette smokers.
Lipid-lowering guidelines endorse a low-density lipoprotein cholesterol goal of <100 mg/dL for people with coronary heart disease (CHD). A more stringent threshold of <70 mg/dL is recommended for those with CHD and "very high-risk" conditions such as diabetes mellitus, metabolic syndrome, or cigarette smoking. Whether chronic kidney disease (CKD) confers a similar risk for recurrent CHD events is unknown. We evaluated the risk for recurrent CHD events and all-cause mortality among 3,938 participants ≥45 years with CHD in the Reasons for Geographic and Racial Differences in Stroke (REGARDS) study. Chronic kidney disease was defined by estimated glomerular filtration rate <60 mL/min per 1.73 m(2) or urinary albumin to creatinine ratio ≥30 mg/g. Participants were categorized by the presence or absence of CKD and any very high-risk condition. Over a median of 4.1 years, the crude incidence (95% CI) of recurrent CHD events were 12.1 (9.0-15.2), 18.9 (15.5-22.3), 35.0 (25.4-44.6), and 34.2 (28.2-40.3) among those without CKD or high-risk conditions; very high-risk conditions alone; and CKD alone and both CKD and very high-risk conditions. After multivariable adjustment, compared with those without CKD or very high-risk conditions, the hazard ratio (95% CI) for recurrent CHD events was 1.45 (1.02-2.05), 2.24 (1.50-3.34), and 2.10 (1.47-2.98) among those with very high-risk conditions alone, CKD alone, and both CKD and very high-risk conditions, respectively. Results were consistent for all-cause mortality. Chronic kidney disease is associated with risk for recurrent CHD events that approximates or is larger than other established very high-risk conditions.


Effect of adenosine triphosphate-sensitive potassium channel openers on lung preservation.
ATP-sensitive potassium channel (KATP) openers have been proven to be involved in ischemic preconditioning, which protects ischemic tissue. However, the effect of KATP openers on ischemia-reperfusion injury of the lungs remains unknown. We investigated whether a KATP opener, pinacidil, can attenuate ischemia-reperfusion injury using an ex vivo rat lung model. Heart-lung blocks were flushed and preserved with phosphate-buffered saline (control group) or with one of the solutions containing pinacidil (pinacidil group) or pinacidil + glibenclamide (a KATP blocker) (glibenclamide group). The control and glibenclamide groups showed significantly higher values with respect to shunt fraction, pulmonary arterial pressure, and peak inspiratory pressure than the pinacidil group. The concentrations of total adenine nucleotides and ATP in the lung after reperfusion became significantly lower in the control and glibenclamide groups than in the fresh group. Lipid peroxidation of the lungs increased significantly in the control and glibenclamide groups after reperfusion. State 3 mitochondrial respiration and State 3/4 ratios of mitochondrial respiration were significantly decreased in the lungs of the control and glibenclamide groups. These findings suggested that KATP openers would maintain the mitochondrial respiratory function during lung preservation, prevent lipid peroxidation after reperfusion, and attenuate ischemia-reperfusion injury.


Effect of a pharmacy-based health literacy intervention and patient characteristics on medication refill adherence in an urban health system.
Limited health literacy is associated with poor understanding of medication instructions and may be related to medication adherence. To evaluate a 3-part intervention (automated telephone reminder calls to refill prescriptions, picture prescription card, and clear health communication training for pharmacists) intended to increase refill adherence through attention to health literacy. Three pharmacies that serve a primarily indigent, minority population served as the intervention sites, and one pharmacy served as the control site. To evaluate the impact of the 6-month intervention on medication adherence, pharmacy refill data were used to calculate the cumulative medication gap (CMG), in which values close to zero indicate better adherence. The primary measure of treatment effect was a comparison of the change in refill adherence within each group, from baseline to follow-up. A total of 173 patients were enrolled in the intervention group and 102 patients in the control group. Medication adherence was significantly different between intervention (CMG = 0.25) and control (CMG = 0.18) groups at baseline (p = 0.004). Refill adherence in the intervention group improved slightly during follow-up (CMG = 0.23), while it worsened slightly in the control group (CMG = 0.21), but the change in adherence between intervention and control groups was not significantly different (p = 0.4). Between 80% and 90% of intervention participants indicated that receiving the picture prescription and phone call interventions helped them remember when and how to take their medications and refill their prescriptions. No patient characteristics, including health literacy, were consistently associated with adherence. Implementation of a 3-part intervention--automated telephone reminders, picture prescription card, and pharmacist communication skills training--did not significantly improve refill adherence among inner-city patients. Further study should explore whether other aspects of medication management, such as dosing or adverse events, can be improved through these types of interventions, implemented either alone or in combination.


The sandflies (Diptera: Psychodidae) in the Turkish province of Hatay: some possible vectors of the parasites causing human cutaneous leishmaniasis.
The province of Hatay, which lies on the Mediterranean coast in south-eastern Turkey, forms an important endemic area for human cutaneous leishmaniasis (HCL). Between the June and November of 2001, the sandflies in eight areas of the province, Erzin, Dortyol, Iskenderun, Kirikhan, Antakya, Samandag, Altinozu and Yayladagi, were surveyed, using sticky-paper traps placed in goat, sheep, cow or chicken sheds close to houses. Although 217 of the 998 sandflies caught belonged to the genus Sergentomyia, the rest were of Phlebotomus species, including some species that, elsewhere, are known to act as vectors of the parasites causing HCL. The Sergentomyia caught were either S. theodori (59.9%) or S. dentata (40.1%). Most of the Phlebotomus were P. sergenti (60.8%) but P. papatasi (18.8%), P. syriacus (9.3%), P. neglectus (5.1%), P. tobbi (2.7%), P. simici (2.2%), P. alexandri (0.9%) and P. kyreniae (0.1%) were also identified (the latter three species for the first time in Hatay province). The identification of the main vector species of sandfly in the province is complicated because it is still uncertain which Leishmania species cause(s) the local HCL. It seems likely, however, that P. sergenti is the main vector in Antakya, Iskenderun, Dortyol, Erzin and Kirikhan. In areas such as Altinozu and Yayladagi, where, although P. sergenti is very rare or absent, HCL cases still occur each year, other species, such as the relatively abundant P. papatasi and P. syriacus, may be the main vectors.


Psychosocial and physical factors associated with lower extremity injury.
With the increasing availability of modern occupant restraints, more drivers and passengers are surviving high-energy crashes. However, a large number, especially those involved in frontal and offset frontal crashes, incur disabling lower extremity injuries. In the past, not much attention was paid to these injuries, as they were usually not life threatening. Despite the low AIS scores associated with injuries to the lower extremities, they pose a major physical and psychological burden on patients' and their ability to return to pre-crash functioning. Associated injuries, such as mild brain injuries, and psychosocial factors such as depression, also influence the long-term outcome.


Semi-exact concentric atomic density fitting: reduced cost and increased accuracy compared to standard density fitting.
A local density fitting scheme is considered in which atomic orbital (AO) products are approximated using only auxiliary AOs located on one of the nuclei in that product. The possibility of variational collapse to an unphysical "attractive electron" state that can affect such density fitting [P. Merlot, T. Kjærgaard, T. Helgaker, R. Lindh, F. Aquilante, S. Reine, and T. B. Pedersen, J. Comput. Chem. 34, 1486 (2013)] is alleviated by including atom-wise semidiagonal integrals exactly. Our approach leads to a significant decrease in the computational cost of density fitting for Hartree-Fock theory while still producing results with errors 2-5 times smaller than standard, nonlocal density fitting. Our method allows for large Hartree-Fock and density functional theory computations with exact exchange to be carried out efficiently on large molecules, which we demonstrate by benchmarking our method on 200 of the most widely used prescription drug molecules. Our new fitting scheme leads to smooth and artifact-free potential energy surfaces and the possibility of relatively simple analytic gradients.


Construction of a high affinity zinc switch in the kappa-opioid receptor.
Very limited structural information is available concerning the superfamily of G-protein-coupled receptors with their seven-transmembrane segments. Recently a non-peptide antagonist site was structurally and functionally replaced by a metal ion site in the tachykinin NK-1 receptor. Here, this Zn(II) site is transferred to the kappa-opioid receptor by substituting two residues at the outer portion of transmembrane V (TM-V), Asp223 and Lys227, and one residue at the top of TM-VI, Ala298, with histidyl residues. The histidyl residues had no direct effect on the binding of either the non-peptide antagonist [3H]diprenorphine or the non-peptide agonist, [3H]CI977, just as these mutations/substitutions did not affect the apparent affinity of a series of other peptide and non-peptide ligands when tested in competition binding experiments. However, zinc ions in a dose-dependent manner prevented binding of both agonist and antagonist ligands with an apparent affinity for the metal ion, which gradually was built up to 10(-6) M. This represents an increase in affinity for the metal ion of about 1000-fold as compared with the wild-type kappa receptor and is specific for Zn(II) as the affinity for e.g. Cu(II) was almost unaffected. The direct transfer of this high affinity metal ion switch between two only distantly related receptors indicates a common overall arrangement of the seven-helix bundle among receptors of the rhodopsin family.


Paircoil2: improved prediction of coiled coils from sequence.
We introduce Paircoil2, a new version of the Paircoil program, which uses pairwise residue probabilities to detect coiled-coil motifs in protein sequence data. Paircoil2 achieves 98% sensitivity and 97% specificity on known coiled coils in leave-family-out cross-validation. It also shows superior performance compared with published methods in tests on proteins of known structure.


Detection of microbial contamination in chicken meat from local markets in Surabaya, East Java, Indonesia.
Chicken meat can be contaminated by microorganisms anywhere in the supply chain, from farm to market, and these microorganisms can be transmitted to humans through direct contact, contact with the environment, and food consumption. The microbial contamination has a serious impact on public health. This study aimed to analyze the microbial contamination of chicken meat sampled from local markets in Surabaya, East Java, Indonesia. A total of 60 samples of fresh chicken meat obtained from 10 traditional markets (six samples per market) were examined for the presence of bacteria. Staphylococcus aureus, Salmonella spp., and Escherichia coli were identified using Gram staining, culturing, and biochemical tests. The most probable number (MPN) method was used to identify E. coli. Most chicken meat samples were positive for S. aureus (58.3%), Salmonella spp. (48.3%), and E. coli (40%). The samples were considered positive for E. coli if the MPN value was higher than 1×101 CFU/g. High microbial contamination was found in all the chicken meat sampled from local markets in Surabaya. Such contamination can lead to foodborne diseases so, proper hygiene and sanitation standards should be followed from slaughterhouses to the end-users.


Disulfide assignment of the C-terminal cysteine knot of agouti-related protein (AGRP) by direct sequencing analysis.
We have assigned the disulfide structure of Md-65 agouti-related protein (Md65-AGRP) using differential reduction and alkylation followed by direct sequencing analysis. The mature human AGRP is a single polypeptide chain of 112 amino acid residues, consisting of an N-terminal acidic region and a unique C-terminal cysteine-rich domain. The C-terminal domain, a 48 amino acid peptide named Md65-AGRP, was expressed in Escherichia coil cells and refolded under different conditions from the mature recombinant protein. The disulfide bonds in the cystine knot structure of Md65-AGRP were partially reduced using tris(2-carboxyethyl) phosphine (TCEP) under acidic conditions, followed by alkylation with N-ethylmaleimide (NEM). The procedure generated several isoforms with varying degrees of NEM alkylation. The multiple forms of Md65-AGRP generated by partial reduction and NEM modification were then completely reduced and carboxymethylated to identify unreactive disulfide bonds. Differentially labeled Md65-AGRP were directly sequenced and analyzed by MALDI mass spectrometry. The results confirmed that Md65-AGRP contained the same disulfide structure as that of Md5-AGRP reported previously [Bures, E. J., Hui, J. O., Young, Y. et al. (1998) Biochemistry 37, 12172-12177].


Delivery of care for hypertension.
To overcome shortcomings in the delivery of care for hypertension various approaches have been developed including a system whereby care of hypertension is shared in a formal manner between general practitioners and hospital specialists. The feasibility, acceptability and cost-effectiveness of a computerised model of shared care were investigated in three matched groups of patients attending hypertension clinics in Glasgow. Glasgow Blood Pressure Clinic attenders considered suitable for shared care by their consultants were randomised to shared care (n = 277) or continued clinic follow-up; a further control group (n = 277) was identified from an independent nurse-practitioner clinic. After 2 years of follow-up, feasibility was estimated by the proportion of patients who had undergone an adequate review (blood pressure, serum creatinine and electrocardiograph); acceptability to general practitioners and patients was assessed by questionnaires; cost-effectiveness was calculated as the cost (to National Health Service and patient) per adequate review. The drop-out rate for shared care over 2 years was 3% compared with 14% for the outpatient clinic and 9% for the nurse-practitioner clinic. In year 2, rates of adequate reviews were 82%, 52% and 75%, respectively. Blood pressure control was similar in the groups. Of 297 general practitioners invited to participate, 85% agreed and 68% wished to continue participation after 2 years. About 50% of shared care patients preferred this method of follow-up compared with their earlier experience of clinic attendance. Shared care was more cost-effective than either conventional or nurse-practitioner clinic follow-up, especially with respect to cost to the patient; costs per adequate review (pound sterling) were 28.96, 50.55 and 30.95, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


V1 interneurons regulate the pattern and frequency of locomotor-like activity in the neonatal mouse spinal cord.
In the mouse spinal cord, V1 interneurons are a heterogeneous population of inhibitory spinal interneurons that have been implicated in regulating the frequency of the locomotor rhythm and in organizing flexor and extensor alternation. By introducing archaerhodopsin into engrailed-1-positive neurons, we demonstrate that the function of V1 neurons in locomotor-like activity is more complex than previously thought. In the whole cord, V1 hyperpolarization increased the rhythmic synaptic drive to flexor and extensor motoneurons, increased the spiking in each cycle, and slowed the locomotor-like rhythm. In the hemicord, V1 hyperpolarization accelerated the rhythm after an initial period of tonic activity, implying that a subset of V1 neurons are active in the hemicord, which was confirmed by calcium imaging. Hyperpolarizing V1 neurons resulted in an equalization of the duty cycle in flexor and extensors from an asymmetrical pattern in control recordings in which the extensor bursts were longer than the flexor bursts. Our results suggest that V1 interneurons are composed of several subsets with different functional roles. Furthermore, during V1 hyperpolarization, the default state of the locomotor central pattern generator (CPG) is symmetrical, with antagonist motoneurons each firing with an approximately 50% duty cycle. We hypothesize that one function of the V1 population is to set the burst durations of muscles to be appropriate to their biomechanical function and to adapt to the environmental demands, such as changes in locomotor speed.


Electrophoresis reliability: I. The contaminant issue.
The effects of the common contaminants--soil, oil, gasoline, salt, acid, base, bleach, and detergent--on various forensically used genetic marker systems were studied. The predicted effects of the various contaminants on the proteins and the electrophoretic separations agreed with the observed results. A contaminant that affected protein conformation also adversely affected the integrity of the electrophoretic system, thus signalling an anomaly. It also was pointed out that the ideal control study for the effects of contaminants on genetic markers in evidentiary material is often provided to forensic scientists--that is victim's blood on victim's clothing and other substrata. The data presented in this paper support the validity and reliability of electrophoretic analyses of evidentiary material with respect to the contaminant issue.


Intersubtype recombinant HIV type 1 involving HIV-MAL-like and subtype H-like sequence in four Norwegian cases.
Suspected epidemiological links between three cases of human immunodeficiency virus type 1 (HIV-1) infection were verified by the finding of a shared unique virus genotype. A probable male index case was not available for testing. Case 1 was a female sexual partner of the index case. Case 2 was an adult son of case 1. Case 3 was a female sexual partner of case 2. The link to the index case was substantiated by the subsequent finding of another female sexual contact of the index case, harboring the same HIV-1 genotype as the three other cases. To characterize the genotype further, the complete provirus nucleotide sequence was obtained directly from blood cell DNA of case 3. HIV cultivated from case 3 demonstrated CCR5 dependence, an extreme slow-low phenotype, and some genotypic features not present in its directly sequenced counterpart. Most of the gag, pol, and vif genes of these viruses clustered with one of the earliest African HIV-1 strains, MAL, previously classified as a recombinant between the subtypes A, D, and I. Most of the rest of the genome was related to subtype H, albeit with less than 90% identity in most regions. These viruses are the only ones shown to display extensive similarity with MAL in the gag-pol region and among the first HIV-1 recombinants described involving subtype H. We postulate that the gag-pol genes of MAL and these viruses are derived from a common ancestor that is not necessarily intersubtype recombinant in the pol region.


Pretreatment pathologic prognostic factors in head and neck squamous cell carcinoma.
Squamous carcinomas of the head and neck region are a common problem for the otolaryngologist; surgeons are obliged to draw together disparate lines of evidence-from physical examination, pathology, and radiology-to plan optimal therapy for their patients. This article explores some of the ways in which pathologic analysis of a biopsy specimen in the past (determination of extent of invasion and degree of differentiation), present (including an analysis of the pattern of infiltration and the tumor' s DNA content), and, perhaps, the future (possibilities including cytogenetics and analysis of discrete steps in the cell cycle) plays a role in this process.


Ligand Selectivity by Inserting GCGC-Tetrads into G-Quadruplex Structures.
G-Quadruplexes (G4s) assembled from tandem G-rich repeat sequences exhibit significant biological functions and applications, which may well depend on their structural features, such as the planar arrangement of G-tetrads and flexibility of loop regions. It has been found that cytosine-intercalated G-repeat sequences also assemble to be quadruplex structures, involving the formation of nonplanar GCGC-tetrads. Herein, to investigate the effect of GCGC-tetrads on structural properties of G4s, some previously studied quadruplexes with or without GCGC-tetrads were selected, and were used to interact with various developed G4 ligands. Our data show that stacked G-tetrads in quadruplexes are important for the π-π stacking interactions, thus promoting the combination with end-stacking ligands, such as porphyrins or planar small molecules. This is confirmed by the observation that the quadruplex formed by d(GGGCT4 GGGC) with two internal G-tetrads and two external GCGC-tetrads can bind to planar ligands in the presence of specific G4-stabilizing cations, including K+ and Pb2+ , and can realize the sensitive detection of Pb2+ . However, the quadruplex composed of two external G-tetrads and two internal GCGC-tetrads formed by d(GCGGT3 GCGG) facilitates the binding of nonplanar ligands, such as triphenylmethane (TPM) dyes, owing to the structural flexibility induced by internal GCGC-tetrads. This work provides new insights into the interaction between DNA quadruplexes and specific ligands, which is beneficial to the development of quadruplex-based biosensors and the design of anticancer drugs.


Management of the crushed chest.
Thoracic injuries are very common among trauma victims. This article reviews the current literature on the management of multiple aspects of the care of the patient with severe chest injury. The mechanics of chest injury are complex and varied. Chest wall injuries are the most common and noticeable manifestation of thoracic trauma. Overall morbidity and mortality are primarily determined by associated injuries. New ventilatory strategies permit oxygenation of the severely hypoxic patient. Acute pain management modalities offer the potential of decreasing associated pulmonary complications. Surgical chest wall fixation is clearly indicated in extreme cases of pulmonary herniation and chest wall disruption. There are potential benefits of surgical fixation in other settings, although further trials are needed.


Prostate carcinoma--the value of T stage and grade in predicting metastases and prognosis. A cost-effective approach to clinical staging.
In order to evaluate the initial T stage and tumour grade as predictors of metastatic disease and prognosis in adenocarcinoma of the prostate, 963 patients were reviewed. Of the patients, 41% presented with metastatic disease. Stage T4 tumours were associated with a consistently poor prognosis, and 70% of such patients had demonstrable distant metastases. No patient with TOf local disease had metastases or died of prostate cancer during follow-up. The incidence of metastases was also low in stages T1 and T2. High tumour grade correlated strongly with more advanced disease. Using this information a more cost-effective approach to the staging of prostate carcinoma is proposed.