Evaluation of the role of Candida albicans agglutinin-like sequence (Als) proteins in human oral epithelial cell interactions.
The fungus C. albicans uses adhesins to interact with human epithelial surfaces in the processes of colonization and pathogenesis. The C. albicans ALS (agglutinin-like sequence) gene family encodes eight large cell-surface glycoproteins (Als1-Als7 and Als9) that have adhesive function. This study utilized C. albicans Δals mutant strains to investigate the role of the Als family in oral epithelial cell adhesion and damage, cytokine induction and activation of a MAPK-based (MKP1/c-Fos) signaling pathway that discriminates between yeast and hyphae. Of the eight Δals mutants tested, only the Δals3 strain showed significant reductions in oral epithelial cell adhesion and damage, and cytokine production. High fungal:epithelial cell multiplicities of infection were able to rescue the cell damage and cytokine production phenotypes, demonstrating the importance of fungal burden in mucosal infections. Despite its adhesion, damage and cytokine induction phenotypes, the Δals3 strain induced MKP1 phosphorylation and c-Fos production to a similar extent as control cells. Our data demonstrate that Als3 is involved directly in epithelial adhesion but indirectly in cell damage and cytokine induction, and is not the factor targeted by oral epithelial cells to discriminate between the yeast and hyphal form of C. albicans.


Studies on infection and immunity in experimental typhoid fever. II. Susceptibility of recovered animals to re-exposure.
Chimpanzees recovered from typhoid fever induced by ingestion of large numbers of phage-type T S. typhosa were rechallenged approximately a year and a half later with a like number of a phage-type E(1) strain. Control animals exhibited febrile responses, bacteriemia, and other significant laboratory and clinical findings, including increases in C-reactive protein levels and sedimentation rates. All of the previously infected and recovered chimpanzees were negative with respect to the aforementioned observations, and appeared to have resisted significant re-infection. On the other hand, recovery from typhoid fever induced by smaller numbers of S. typhosa failed to protect completely against rechallenge with larger numbers of these organisms. These findings indicate that chimpanzees recovered from typhoid fever may be protected against re-infection, even though the re-infecting organism is of a different phage type. Differences in the magnitude of the original and subsequent infecting inocula appear to influence the response to rechallenge.


Prevention and management of malaria during pregnancy: findings from a comparative qualitative study in Ghana, Kenya and Malawi.
In endemic regions of sub-Saharan Africa, malaria during pregnancy (MiP) is a major preventable cause of maternal and infant morbidity and mortality. Current recommended MiP prevention and control includes intermittent preventive treatment (IPTp), distribution of insecticide-treated bed nets (ITNs) and appropriate case management. This article explores the social and cultural context to the uptake of these interventions at four sites across Africa. A comparative qualitative study was conducted at four sites in three countries: Ghana, Malawi and Kenya. Individual and group interviews were conducted with pregnant women, their relatives, opinion leaders, other community members and health providers. Observations, which focused on behaviours linked to MiP prevention and treatment, were also undertaken at health facilities and in local communities. ITNs were generally recognized as important for malaria prevention. However, their availability and use differed across the sites. In Malawi and Kenya, ITNs were sought-after items, but there were complaints about availability. In central Ghana, women saved ITNs until the birth of the child and they were used seasonally in northern Ghana. In Kenya and central Ghana, pregnant women did not associate IPTp with malaria, whereas, in Malawi and northern Ghana, IPTp was linked to malaria, but not always with prevention. Although IPTp adherence was common at all sites, whether delivered with directly observed treatment or not, a few women did not comply with IPTp often citing previous side effects. Although generally viewed as positive, experiences of malaria testing varied across the four sites: treatment was sometimes administered in spite of a negative diagnosis in Ghana (observed) and Malawi (reported). Despite generally following the advice of healthcare staff, particularly in Kenya, personal experience, and the availability and accessibility of medication--including anti-malarials--influenced MiP treatment. Although ITNs were valued as malaria prevention, health messages could address issues that reduce their use during pregnancy in particular contexts. The impact of previous side effects on adherence to IPTp and anti-malarial treatment regimens during pregnancy also requires attention. Overtreatment of MiP highlights the need to monitor the implementation of MiP case management guidelines.


Evidence for tautomerism in nucleic acid base pairs. 1H NMR study of 15N labeled tRNA.
The imino proton resonances of 15N labeled tRNA appear as asymmetric doublet signals, the asymmetry being dependent on the applied magnetic field strength. Assuming a tautomerism of the type N-H...N not equal to N...H-N in the base pairs the line shapes can be simulated. The most important parameters fitted in the simulation are the rate constants of the proton transfer and the mole fractions of either tautomeric state. The rate constants are of the order of 100s-1 and the mole fractions of the non dominant tautomer about 0.1 depending on the temperature and on the nature of the base pairing. The observations are attributed to a double proton transfer in the base pairs. The unexpectedly slow rates of the double proton transfer process may be connected with a concomitant conformational change of the duplex structure.


Influence of fluorinated graphene-modified epoxy coatings on the corrosion behaviour of 2024 aluminium alloy.
This study provides an enhanced corrosion resistance of epoxy resin (EP) by embedding fluorinated graphene (FG) into the epoxy matrix. FG with different fluorine contents was obtained by reacting nitrogen trifluoride (NF3) gas with GO and then incorporated into the EP matrix to fabricate the different composites. Through a series of characterization methods, the chemical composition and microstructures of FG were systematically analyzed, and its corrosion resistance was also studied. Results revealed that F atoms were bonded to the GO surface to form C-F covalent bonds, and an FG lamellar thickness less than 2 nm. The contact angle of the coatings increased with the incorporation of FG, and the coating resistance of FG2/EP coating was 3 orders of magnitude more than that of the EP coating after immersion for 4080 h. Thus, the incorporation of FG into epoxy matrix significantly enhanced its hydrophobic properties and barrier performance, which was beneficial to improving the long-term corrosion resistance of the coating.


One-pot catalytic enantio- and diastereoselective syntheses of anti-, syn-cis-disubstituted, and syn-vinyl cyclopropyl alcohols.
Highly enantio- and diastereoselective methods for the synthesis of a variety of cyclopropyl alcohols are reported. These methods represent the first one-pot approaches to syn-vinyl cyclopropyl alcohols, syn-cis-disubstituted cyclopropyl alcohols, and anti-cyclopropyl alcohols from achiral precursors. The methods begin with enantioselective C-C bond formations promoted by a MIB-based zinc catalyst to generate allylic alkoxide intermediates. The intermediates are then subjected to in situ alkoxide-directed cyclopropanation to provide cyclopropyl alcohols. In the synthesis of vinyl cyclopropyl alcohols, hydroboration of enynes is followed by transmetalation of the resulting dienylborane to zinc to provide dienylzinc reagents. Enantioselective addition to aldehydes generates the requisite dienyl zinc alkoxides, which are then subjected to in situ cyclopropanation to furnish vinyl cyclopropyl alcohols. Cyclopropanation occurs at the double bond allylic to the alkoxide. Using this method, syn-vinylcyclopropyl alcohols are obtained in 65-85% yield, 76-93% ee, and > 19:1 dr. To prepare anti-cyclopropanols, enantioselective addition of alkylzinc reagents to conjugated enals provides allylic zinc alkoxides. Because direct cyclopropanation provides syn-cyclopropyl alcohols, the intermediate allylic alkoxides were treated with TMSCl/Et(3)N to generate intermediate silyl ethers. In situ cyclopropanation of the allylic silyl ether resulted in cyclopropanation to form the anti-cyclopropyl silyl ether. Workup with TBAF affords the anti-cyclopropyl alcohols in one pot in 60-82% yield, 89-99% ee, and > or = 10:1 dr. For the synthesis of cis-disubstituted cyclopropyl alcohols, in situ generated (Z)-vinyl zinc reagents were employed in asymmetric addition to aldehydes to generate (Z)-allylic zinc alkoxides. In situ cyclopropanation provides syn-cis-disubstituted cyclopropyl alcohols in 42-70% yield, 88-97% ee, and > 19:1 dr. These one-pot procedures enable the synthesis of a diverse array of cyclopropyl alcohol building blocks with high enantio- and diastereoselectivities.


Characteristics and outcomes of pediatric rapid response teams before and after mandatory triggering by an elevated Pediatric Early Warning System (PEWS) score.
The Pediatric Early Warning System (PEWS) was created to identify unstable patients before their deterioration. Rapid response teams (RRTs) were developed to assist with management of such patients. In 2009, our institution mandated the activation of RRTs if a PEWS score was elevated (ie, ≥5). The goal of this study was to examine changes in characteristics of RRT calls before and after the implementation of a mandatory hospital policy requiring RRT activation due to an elevated PEWS score. This study was a retrospective database review, with RRT data from June 2007 to December 2010 examined. A total of 44 RRTs were recorded before mandatory triggering and 69 RRTs afterward in the study period (P = .32). Compared with the premandatory group, the mandatory triggering group found that tachycardia was a more frequent trigger for RRTs, with an increase of 26.1% (P = .004). RRTs triggered by a change in mental status/agitation decreased by 22.9% (P = .009). An increase of 15.1% of RRTs required no interventions with mandatory triggering. Nighttime RRTs increased by17.5% (P = .07). There was a trend toward decreased PICU transfers in the mandatory triggering group, with no significant change in code blue calls. A hospital policy of mandating RRT activation based on PEWS scores increased nighttime calls and altered the primary reasons for RRT activation in our center, with no evidence of improvements in patient care. These findings should be interpreted with caution given the relatively rare outcomes the policy is intended to prevent; however, our findings highlight the difficulties inherent in evaluating methods to improve pediatric patient safety.


Minimally Invasive Techniques for Iliac Bolt Placement: 2-Dimensional Operative Video.
As the popularity of minimally invasive surgery (MIS) continues to grow, novel techniques are needed to meet the demands of multisegment fixation for advanced spinal diseases. In one such example, iliac bolts are often required to anchor large fusion constructs, but MIS technical notes are missing from the literature. A 67-yr-old female presented with a symptomatic coronal deformity: preoperative pelvic incidence = 47°, pelvic tilt = 19°, and lumbar lordosis = 29°, sagittal vertical axis = +5.4 cm with 30° of scoliosis. The operative plan included T10-ilium fusion with transforaminal interbody grafts at L2-3, L3-4, L4-5, and L5-S1. The intraoperative video is of minimally invasive placement of iliac bolts using the O-Arm Surgical Imaging System (Medtronic®). The patient consented to the procedure. A mini-open exposure that remains above the fascial planes allows for multilevel instrumentation with appropriate decompression at the interbody segments. After the placement of the pedicle screws under image-guidance, the direction is turned to the minimally invasive iliac bolts. Following the trajectory described in the standard open approach,1 the posterior superior iliac spine (PSIS) is identified with the navigation probe, which will guide the Bovie cautery through the fascia. This opening assists in the trajectory of the navigated-awl tap toward the anterior superior iliac spine (ASIS). Next, 8.5 mm x 90 mm iliac screws were placed in the cannulated bone under navigation. After intraoperative image confirmation of screw placement, the contoured rods are threaded under the fascia. The setscrews lock the rod in position. MIS approaches obviate cross-linking the rods, rendering pelvic fixation more facile. This technique allows for minimal dissection of the posterior pelvic soft tissue while maintaining adequate fixation.


Acetylcholinesterase active centre and gorge conformations analysed by combinatorial mutations and enantiomeric phosphonates.
A series of eight double and triple mutants of mouse acetylcholinesterase (AChE; EC 18.104.22.168), with substitutions corresponding to residues found largely within the butyrylcholinesterase (BChE; EC 188.8.131.52) active-centre gorge, was analysed to compare steady-state kinetic constants for substrate turnover and inhibition parameters for enantiomeric methylphosphonate esters. The mutations combined substitutions in the acyl pocket (Phe(295)-->Leu and Phe(297)-->Ile) with the choline-binding site (Tyr(337)-->Ala and Phe(338)-->Ala) and with a side chain (Glu(202)--> Gln) N-terminal to the active-site serine, Ser(203). The mutations affected catalysis by increasing K (m) and decreasing k (cat), but these constants were typically affected by an order of magnitude or less, a relatively small change compared with the catalytic potential of AChE. To analyse the constraints on stereoselective phosphonylation, the mutant enzymes were reacted with a congeneric series of S (P)- and R (P)-methylphosphonates of known absolute stereochemistry. Where possible, the overall reaction rates were deconstructed into the primary constants for formation of the reversible complex and intrinsic phosphonylation. The multiple mutations greatly reduced the reaction rates of the more reactive S (P)-methylphosphonates, whereas the rates of reaction with the R (P)-methylphosphonates were markedly enhanced. With the phosphonates of larger steric bulk, the enhancement of rates for the R (P) enantiomers, coupled with the reduction of the S (P) enantiomers, was sufficient to invert markedly the enantiomeric preference. The sequence of mutations to enlarge the size of the AChE active-centre gorge, resembling in part the more spacious gorge of BChE, did not show an ordered conversion into BChE reactivity as anticipated for a rigid template. Rather, the individual aromatic residues may mutually interact to confer a distinctive stereospecificity pattern towards organophosphates.


Nitric oxide and sphingolipids: mechanisms of interaction and role in cellular pathophysiology.
Nitric oxide is a short-lived messenger with pleiotropic roles in the regulation of cell patho-physiological processes, including survival, death, proliferation and differentiation. Increasing evidence over the last few years has shown that nitric oxide effects in apoptosis, growth and differentiation originate in significant part from its interplay with signalling members of the sphingolipid family. In many cell types belonging to different lineages, nitric oxide and sphingolipids interact in two-way pathways leading to regulation of the activity and expression of enzymes involved in each other's signalling events. These crosstalk signalling events involve various sphingolipids, with key roles for ceramide and sphingosine-1-phosphate, and signal transduction molecules downstream of nitric oxide, with cyclic GMP as a main player. The biological implications of some of these interactions are now being understood. The best-characterised so far, the mutual regulation of sphingomyelinases and endothelial nitric oxide synthase, acts as a tuning system in crucial patho-physiological processes such as inflammation, proliferation and cell death.


Characterization of Triatoma virus, a picorna-like virus isolated from the triatomine bug Triatoma infestans.
Some properties of Triatoma virus (TrV), a picorna-like virus recently isolated from Triatoma infestans, have been studied. Electron microscopic observations of purified viral preparations showed the presence of non-enveloped viral particles 30 nm in diameter. The sedimentation coefficient of virus particles was about 165S and the buoyant density in CsCl was 1.39 g/ml. The viral genome was composed of one single-stranded RNA molecule with an Mr of 3 x 10(6). Three major polypeptides with Mr values of 39K, 37K and 33K and a minor one of about 45K were found in the virus particle. TrV particles contain about 35% RNA and 65% protein by weight. These data support the classification of this virus in the family Picornaviridae.


Kinematic characteristics of standing disequilibrium: reliability and validity of a posturographic protocol.
To investigate retest reliability and concurrent validity of the fundamental measurements made of a posturographic protocol that employs quiet standing to quantify the severity and the nature of patients' postural disturbances. Retrospective complete block design. Geriatric rehabilitation department. Thirty-six participants (age range, 67 to 86 yrs) having normal, moderate, or severe levels of disequilibrium. Quiet standing was evaluated on three occasions using a three-dimensional motion analysis system and a force platform. Eight testing conditions, designed to vary task difficulty by controlling the contributions of vision, foot proprioception, and base-of-support width, were administered. Retest reliability of body sway, joint alignment, body position, and motor coordination indicators were evaluated by intraclass correlation coefficients (ICCs). Concurrent validity of protocol measures was evaluated by the prediction of disequilibrium from a stepwise linear discriminant analysis. ICCs indicated high level of retest reliability for all variables but those of motor coordination, which was not influenced by testing conditions. Discriminant analysis resulted in a four-factor discriminator that included measures of body sway, position, alignment, and motor coordination. The derived linear discriminate function correctly classified 96% of the patients' level of disequilibrium. The posturographic protocol has the potential to be a useful tool for evaluating severity and nature of postural instability and the effects of pharmacologic and rehabilitative treatment. Results also indicate that combining direct body measurements with force-plate data has the potential to expose the underlying impairments that cause disequilibrium, determine their pathogenesis, and evaluate compensatory strategies.


Dental fragment embedded in the lower lip after facial trauma: Brief review literature and report of a case.
Upper incisors are the most frequently involved teeth in traumatic dental injuries. Soft tissues (lips and/or oral mucosa) adjacent to incisal edge can receive direct and/or indirect traumas. Laceration of the lower lip is a not rare eventuality and teeth fragments could be embedded in labial soft tissue. The reattachment of these fragments, if possible, is the elective treatment choice, thanks to the modern adhesive and restorative techniques. The authors present a case of a white Caucasian 10-year-old child, who attended the dental clinic for the treatment of both upper central incisors' crown fractures. The fragment of the left incisor was retrieved embedded in the lower lip. It was successfully surgically removed and reattached using a composite adhesive technique. A careful clinical and radiographic examination with the surgical removal of tooth fragments could prevent undesirable foreign body reaction, infection and scarring. The authors also reviewed the most relevant literature concerning tooth fragment reattachment after removal from oral soft tissues.


Physicochemical and microbiological characteristics of kitoza, a traditional salted/dried/smoked meat product of Madagascar.
Kitoza samples collected from producers in Madagascar were analyzed for their physicochemical and microbial properties. Lactic acid bacteria and coagulase-negative staphylococci were the two codominant populations with average counts of 6-7 log cfu/g. Good hygienic practices were sometimes lacking but samples were not contaminated with Salmonella, Clostridium perfringens, and Bacillus cereus and only once with Listeria monocytogenes. Staphylococcus aureus was found occasionally with higher counts in salted/dried products than in salted/smoked products. Moisture, protein, fat, and salt contents varied considerably and were on average 41.5, 43.5, 14.3, and 3.3 g/100 g, respectively, and water activity was 0.893 on average. Smoked kitoza showed higher moisture content compared to dried kitoza. Most of the smoked kitoza had a water activity higher than 0.9 which is not in accordance with their storage at ambient temperatures. Benzo(a)pyrene content was above 2 µg/kg in 11 out of 30 smoked samples (17 ± 16.5 µg/kg on average).


Respiratory effects from industrial talc exposure among former mining workers.
To evaluate late respiratory effects from occupational inhalation of talc contaminated with asbestos. This was a case series study on 29 former talc mining workers with asbestos contamination, in the municipality of Carandaí, Southeastern Brazil, who were attended at the State Workers' Health Reference Center in 2004 and 2005. Their clinical and occupational histories were obtained and they underwent spirometry and chest radiography. An exposure score was created; multiplying this by duration produced a cumulative talc exposure index. To confirm the association between the cumulative exposure index and the presence of radiological abnormalities suggestive of pneumoconiosis and/or pleural abnormalities, an exact logistic regression model was fitted to this. All the former workers were males, with an average age of 48.2 years. Chest radiographs showed pleural abnormalities in three of them; parenchymatous opacity compatible with pneumoconiosis in one; and suspected pneumoconiosis in six. Spirometric abnormalities were found in three workers. Logistic regression showed an odds ratio of 1.059 (95% CI: 1.012; 1.125) for the cumulative exposure index, i.e. each unit increase in the index resulted in an increase of 5.9% in the chance of presenting radiological abnormalities compatible with or suspicious of pneumoconiosis. With regard to the median estimated latency period between the start of exposure and the diagnosing of pleural plaque, there was a significant difference (p = 0.013) between the cases (27.0 years) and non-cases (14.3 years). These findings indicate the need for clinical control among workers who have been exposed to asbestos, particularly because of the late effects from exposure to this mineral.


Soil and water contamination with carbofuran residues in agricultural farmlands in Kenya following the application of the technical formulation Furadan.
This study was undertaken to determine the concentrations of carbofuran residues in water, soil and plant samples from selected sites in the farmlands in Kenya and to demonstrate the impact of Furadan use on the local environment. Soil, water and plant samples obtained from agricultural farmlands where the technical formulation Furadan has been used extensively showed high environmental contamination with concentrations of carbofuran and its two toxic metabolites 3-hydroxycarbofuran and 3-ketocarbofuran, separately, ranging from 0.010-1.009 mg/kg of dry surface soil, 0.005-0.495 mg/L in water samples from two rivers flowing through the farms and bdl-2.301 mg/L in water samples from ponds and dams located close to the farms. Maize plant samples contained these residues in concentrations ranging from 0.04-1.328 mg/kg of dry plant tissue. The significantly high concentration levels of carbofuran and its metabolites, 3-ketocarbofuran and 3-hydroxycarbofuran, found in various matrices demonstrate that Furadan was used extensively in the two areas and that there was environmental distribution and exposure of residues in water which posed risks when used for domestic purposes or as drinking water for animals in two wildlife conservancies where the dams and ponds are located. Surface soil contamination was also high and posed risks through run-off into the dams and rivers as well as through secondary exposure to small birds and mammals.


Design of computer-generated beam-shaping holograms by iterative finite-element mesh adaption.
Computer-generated phase-only holograms can be used for laser beam shaping, i.e., for focusing a given aperture with intensity and phase distributions into a pregiven intensity pattern in their focal planes. A numerical approach based on iterative finite-element mesh adaption permits the design of appropriate phase functions for the task of focusing into two-dimensional reconstruction patterns. Both the hologram aperture and the reconstruction pattern are covered by mesh mappings. An iterative procedure delivers meshes with intensities equally distributed over the constituting elements. This design algorithm adds new elementary focuser functions to what we call object-oriented hologram design. Some design examples are discussed.


diseaseGPS: auxiliary diagnostic system for genetic disorders based on genotype and phenotype.
The next-generation sequencing brought opportunities for the diagnosis of genetic disorders due to its high-throughput capabilities. However, the majority of existing methods were limited to only sequencing candidate variants, and the process of linking these variants to a diagnosis of genetic disorders still required medical professionals to consult databases. Therefore, we introduce diseaseGPS, an integrated platform for the diagnosis of genetic disorders that combines both phenotype and genotype data for analysis. It offers not only a user-friendly GUI web application for those without a programming background but also scripts that can be executed in batch mode for bioinformatics professionals. The genetic and phenotypic data are integrated using the ACMG-Bayes method and a novel phenotypic similarity method, to prioritize the results of genetic disorders. diseaseGPS was evaluated on 6085 cases from Deciphering Developmental Disorders project and 187 cases from Shanghai Children's hospital. The results demonstrated that diseaseGPS performed better than other commonly used methods. diseaseGPS is available to freely accessed at https://diseasegps.sjtu.edu.cn with source code at https://github.com/BioHuangDY/diseaseGPS.


Point sources of air pollution.
Many people live near point sources of air pollution such as industrial sites and waste disposal operations and there are often suggestions of clusters of disease around such activities. Such alleged clusters will generate significant public concern and media interest and in many cases will warrant detailed investigation. However, the ability of current epidemiological methods to investigate such clusters is limited, particularly with regard to obtaining reliable and accurate population exposure data. In many cases, the key question is whether releases from a point source result in a significant increase in exposure or whether other sources (background exposure) give rise to the dominant exposure. This review considers some of the issues around point sources including methods of estimating exposure and briefly discusses some of the epidemiological evidence linking respiratory disease and cancer with specific industries such as coking works and incinerators.


Virus-tumor synergism.
More than 30 mouse tumors are associated with virus-like agents that may readily be detected by enzymic techniques. Radiation and chemically induced tumors do not ordinarily give evidence of such activity. The present report is of experiments in which a synergistic effect has been observed to occur when animals were inoculated with both the filtrable agent and a virus-free tumor. Synergism was shown by accelerated growth of the tumor and by elevation of a glycolytic enzyme (lactic dehydrogenase) in the host plasma


Nuclear Lamin B1 Interactions With Chromatin During the Circadian Cycle Are Uncoupled From Periodic Gene Expression.
Many mammalian genes exhibit circadian expression patterns concordant with periodic binding of transcription factors, chromatin modifications, and chromosomal interactions. Here we investigate whether chromatin periodically associates with nuclear lamins. Entrainment of the circadian clock is accompanied, in mouse liver, by a net gain of lamin B1-chromatin interactions genome-wide, after which the majority of lamina-associated domains (LADs) are conserved during the circadian cycle. By tailoring a bioinformatics pipeline designed to identify periodic gene expression patterns, we also observe hundreds of variable lamin B1-chromatin interactions among which oscillations occur at 64 LADs, affecting one or both LAD extremities or entire LADs. Only a small subset of these oscillations however exhibit highly significant 12, 18, 24, or 30 h periodicity. These periodic LADs display oscillation asynchrony between their 5' and 3' borders, and are uncoupled from periodic gene expression within or in the vicinity of these LADs. Periodic gene expression is also unrelated to variations in gene-to-nearest LAD distances detected during the circadian cycle. Accordingly, periodic genes, including central clock-control genes, are located megabases away from LADs throughout circadian time, suggesting stable residence in a transcriptionally permissive chromatin environment. We conclude that periodic LADs are not a dominant feature of variable lamin B1-chromatin interactions during the circadian cycle in mouse liver. Our results also suggest that periodic hepatic gene expression is not regulated by rhythmic chromatin associations with the nuclear lamina.


Pruned tree-structured vector quantization of medical images with segmentation and improved prediction.
The authors use predictive pruned tree-structured vector quantization for the compression of medical images. Their goal is to obtain a high compression ratio without impairing the image quality, at least so far as diagnostic purposes are concerned. The authors use a priori knowledge of the class of images to be encoded to help them segment the images and thereby to reserve bits for diagnostically relevant areas. Moreover, the authors improve the quality of prediction and encoding in two additional ways: by increasing the memory of the predictor itself and by using ridge regression for prediction. The improved encoding scheme was tested via computer simulations on a set of mediastinal CT scans; results are compared with those obtained using a more conventional scheme proposed recently in the literature. There were remarkable improvements in both the prediction accuracy and the encoding quality, above and beyond what comes from the segmentation. Test images were encoded at 0.5 bit per pixel and less without any visible degradation for the diagnostically relevant region.


Hitting multiple targets with multimeric ligands.
Multimeric ligands consist of multiple monomeric ligands attached to a single backbone molecule, creating a multimer that can bind to multiple receptors or targets simultaneously. Numerous examples of multimeric binding exist within nature. Due to the multiple and simultaneous binding events, multimeric ligands bind with an increased affinity compared to their corresponding monomers. Multimeric ligands may provide opportunities in the field of drug discovery by providing enhanced selectivity and affinity of binding interactions, thus providing molecular-based targeted therapies. However, gaps in our knowledge currently exist regarding the quantitative measures for important design characteristics, such as flexibility, length and orientation of the inter-ligand linkers, receptor density and ligand sequence. In this review, multimeric ligand binding in two separate phases is examined. The prerecruitment phase describes the binding of one ligand of a multimer to its corresponding receptor, an event similar to monomeric ligand binding. This results in transient increases in the local concentration of the other ligands, leading to apparent cooperativity. The postrecruitment phase only occurs once all receptors have been aligned and bound by their corresponding ligand. This phase is analogous to DNA-DNA interactions in that the stability of the complex is derived from physical orientation. Multiple factors influence the kinetics and thermodynamics of multimeric binding, and these are discussed.


A Molecular Dual Pump.
Artificial molecular machines (AMMs) built from mechanically interlocked molecules (MIMs) can use energy ratchets to control the unidirectional motion of their component parts. These energy ratchets are operated by the alteration of kinetic barriers and thermodynamic wells, which are, in turn, determined by the switching on and off of noncovalent interactions. Previously, we have developed artificial molecular pumps (AMPs) capable of pumping rings consecutively onto a collecting chain as part of a molecular dumbbell, leading to the formation of rotaxanes. Here, we report a molecular dual pump (MDP) consisting of two individual AMPs linked in a head-to-tail fashion, wherein a single ring is pumped, in a linear manner, on and off a dumbbell involving a [2]rotaxane intermediate by exploiting the redox properties of the two pumps. This MDP, defined by the finely tuned noncovalent interactions and fueled by either chemicals or electricity, utilizes an energy ratchet mechanism to capture a ring and subsequently release it back into solution. The unidirectional motion and the resulting controlled capture and release of the ring were followed by 1D and 2D 1H NMR spectroscopy and supported by control experiments. This molecular dual pump may be considered to be a forerunner of AMMs that are capable of pumping rings across a membrane in a way similar to how bacteriorhodopsin transports protons from one side of a membrane to the other under the influence of light. Such extensive multicomponent AMMs can lead potentially to molecular transporting platforms with positional and directional control of cargo uptake and release when, and only when, instructed.


Incidence of refractory CMV infection with or without antiviral resistance in Spain: A systematic literature review.
Solid organ transplantation (SOT) and hematopoietic stem cell transplantation (HSCT) recipients are susceptible to cytomegalovirus (CMV) infection. The incidence of refractoriness to antivirals, with or without resistance, is unclear. The purpose of this review was to describe the epidemiology of refractory CMV infection in Spain to understand the current unmet needs. PubMed, EMBASE, Cochrane and MEDES were searched systematically for relevant articles. We included randomized controlled trials and observational studies published during the period from January 1990 to June 2021. From 212 screened records, we selected 19 papers including 1973 transplant recipients. Refractory infection ranged from 3 to 10% in studies with SOT recipients. The incidence of CMV resistance ranged from 1% to 36% in these patients. The incidence of CMV refractory infection in HSCT recipients ranged from 11 to 50%, while values for resistant infection ranged from 0% to 21%. The wide range of definitions and values observed does not allow us to establish the true incidence of refractory CMV infection with or without resistances in SOT and HSCT patients in Spain. This review highlights the gap between clinical practice and clinical trials' definitions which needed to be updated to be easier followed in current clinical practice.


New approach for correcting distortions in echo planar imaging.
A new method is described that can correct the distortions due to multiple off-resonance effects in echo planar imaging, including those caused by B(0) field inhomogeneities, eddy currents, and gradient waveform imperfections. The proposed method uses a phase encoded acquisition and is as effective as the method of Chen and Wyricz (Chen and Wyricz, Magn Reson Med 1999;41:1206-1213) in correcting for distortions. Unlike Chen and Wyricz's approach, this new method works directly in distorted space and requires fewer scans. It also avoids the difficulties of phase unwrapping inherent in field mapping methods. Results using this new method with phantoms and human head scans at 3.0 T demonstrate the efficacy of the method in correcting distortions in both spin echo echo planar imaging (EPI) and gradient echo EPI.


Chronic exposure to a 60-Hz electric field: effects on synaptic transmission and peripheral nerve function in the rat.
Several reports have suggested that the nervous system can be affected by exposure to electric fields and that these effects may have detrimental health consequences for the exposed organism. The purpose of this study was to investigate the effects of chronic (30-day) exposure of rats to a 60Hz, 100-kV/m electric field on synaptic transmission and peripheral-nerve function. One hundred forty-four rats, housed in individual polycarbonate cages were exposed to uniform, vertical, 60-Hz electric fields in a system free of corona discharge and ozone formation and in which the animals did not receive spark discharges or other shocks during exposure. Following 30 days of exposure to the electric field, superior cervical sympathetic ganglia, vagus and sciatic nerves were removed from rats anesthetized with urethan, placed in a temperature-controlled chamber, and superfused with a modified mammalian Ringer's solution equilibrated with 95% O2 and 5% CO2. Several measures and tests were used to characterize synaptic transmission and peripheral-nerve function. These included amplitude, area, and configuration of the postsynaptic or whole-nerve compound-action potential; conduction velocity; accommodation; refractory period; strength-duration curves; conditioning-test (C-T) response, frequency response; post-tetanic response; and high-frequency-induced fatigue. The results of a series of neurophysiologic tests and measurements indicate that only synaptic transmission is significantly and consistently affected by chronic (30-day) exposure to a 60-Hz, 100-kV/m electric field. Specifically, and increase in synaptic excitability was detected in replicated measurements of the C-T response ratio. In addition, there are trends in other data that can be interpreted to suggest a generalized increase in neuronal excitability in exposed animals.


[Organization of lung digital X-ray fluorography and its introduction into the health care services of Russia].
Based on comprehensive study of the capacities of digital X-ray fluorography (DXRF), the authors give their views on two main aspects of its use. These are firstly the most acceptable organizational forms of lung DXRF into the health care services of Russia and, secondly, its place in the diagnosis of pulmonary tuberculosis, lung cancer, chronic nonspecific lung diseases, and mediastinal tumors. As for its organization and introduction, the authors are against the fact that film fluorography should be rapidly replaced by DXRF. They give recommendations based on the reasonable and stepwise introduction of DXRF, which may rule out their negative perception versus the present attitude to the so-called prophylactic film fluorography. In the part dealing with diagnosis that analyzes 955 cases of various lung diseases (from over 8000 DXRFs) verified by different studies (morphological, cytological, etc.) and chest X-ray computed tomography, the authors show the results of monitoring imaging, by demonstrating all its potentialities. At the same time they recommend that image printing fixing should be also rather widely employed. In the authors' opinion, DXRF may become the method of choice in detecting pulmonary tuberculosis, lung cancer, chronic nonspecific lung diseases, and mediastinal tumors.


Right ventricular echocardiographic indices predict poor outcomes in infants with persistent pulmonary hypertension of the newborn.
Infants with persistent pulmonary hypertension of the newborn (PPHN) have elevated pulmonary vascular resistance that can lead to right ventricular (RV) failure and death. Clinicians must decide which infants will fail conventional therapy and require transfer to extra corporeal membrane oxygenation (ECMO) centres, but accurate echocardiographic predictors have not been identified. We assessed echocardiographic measurements of RV pressure and function in predicting progression to death or ECMO in infants with PPHN. Echocardiograms for infants ≥35-week gestation with a clinical diagnosis of PPHN were retrospectively reviewed. Traditional and strain echocardiographic measures were compared for those with or without the primary outcome of ECMO/cardiovascular death. Receiver operator curves identified cut points for measures that were significantly different. Of the 86 subjects analysed, 25 (29%) of the patients had the primary outcome of ECMO/death. The ECMO/death group had diminished tricuspid annular plane systolic excursion (TAPSE; P = 0.002) and RV global longitudinal peak strain (GLPS; P = 0.03), a predominant right-to-left shunt across the patent ductus arteriosus (PDA; P = 0.05), and an elevated oxygenation index (OI; P < 0.001). Sensitivity/specificity for TAPSE <4 mm was 56 and 85%, and for GLPS greater than or equal to -9% was 52 and 77%. TAPSE, GLPS, and right-to-left PDA shunting were associated with progression to death/ECMO. RV free wall strain was not associated with the outcome, suggesting that diminished global strain better reflects clinical outcomes in this group. These thresholds may assist in the decision-making to transfer high-risk infants to ECMO centres.


Combined effects of nocturnal exposure to artificial light and habitat complexity on fish foraging.
Due to the widespread use of artificial light, freshwater ecosystems in urban areas at night are often subjected to light of intensities exceeding that of the moonlight. Nocturnal dim light could modify fish behaviour and benefit visual predators because of enhanced foraging success compared to dark nights. However, effects of nocturnal light could be mitigated by the presence of structured habitats providing refuges for prey. We tested in laboratory experiments whether nocturnal light of low intensity (2 lx) increases foraging efficiency of the Eurasian perch (Perca fluviatilis) on invertebrate prey (Gammarus fossarum). The tests were conducted at dusk and night under two light regimes: natural cycle with dark nights and disturbed cycle with artificially illuminated nights, in habitats differing in structural complexity: sand and woody debris. We found that nocturnal illumination significantly enhanced the consumption of gammarids by fish compared to dark nights. In addition, the perch was as effective predator in illuminated nights (2 lx) as at dusk (10 lx). Woody debris provided an effective refuge only in combination with undisturbed darkness, but not in illuminated nights. Our results suggest that nocturnal illumination in aquatic ecosystems may contribute to significant reductions in invertebrate population sizes through fish predation. The loss of darkness reduces the possibility of using shelters by invertebrates and hence the effects of elevated light levels at night could not be mitigated by an increased habitat complexity.


The impact of multiple sclerosis on the identity of mothers in Italy.
This paper reports on one of the themes that emerged from the analysis of the study, regarding the perceived influence of multiple sclerosis (MS) on the identity of mothers in the socio-cultural context of Italy. In-depth interviews were conducted with 16 women at various stages of MS, with follow up interviews with seven of the women. Phenomenology guided the methodology and the analysis was conducted using interpretative phenomenological analysis. Through the research the value of motherhood to the women who participated emerged. The findings illustrated how many strove to maintain control of their MS, which led to some making comparisons of themselves and other mothers and feeling different. Some women described how they adjusted their roles and found strength in being mothers but others spoke of their feelings of loss. Most women described living in the moment, appreciating the present and living each day as it came. Another significant experience was fear of stigma, both realized in the form of "pity" from others, and the perceived and actual associated stigma for their families. This contributed to why some women were reluctant to disclose their condition. The mothers who took part in this study differed in how they perceived their disabled identity. Although this study was conducted in the socio-cultural setting of Italy, the findings have implications for professionals working with disabled mothers and women with MS in Italy and beyond; including recognizing the value associated with fully identifying oneself as a mother, rather than solely focusing on doing mothering tasks. Implications for Rehabilitation Professionals need to be mindful of the value of motherhood for women with multiple sclerosis. Professionals should support women who feel like they are battling with maintaining control of their multiple sclerosis, who may be adjusting their identity as mothers; recognizing that they may be influenced by the stage of their multiple sclerosis and whether they were diagnosed before or after having their children. Women can have feelings of loss related to their ability to fully participate in their children's lives and professionals should work with women to help them identify the value of their mothering role not only in physically participating in activities but also in being emotionally and physically present as a mother.


Mechanism of early and late polykaryocytosis induced by the bovine leukaemia virus.
Syncytia formation induced by the bovine leukaemia virus (BLV) has been classified as 'early' or 'late' polykaryocytosis. Early polykaryocytosis arises in the first 24 h in mixed cultures of BLV-infected cells with indicator cells. Late polykaryocytosis is observed 4 to 8 days after infection of sensitive cells with cell-free infectious BLV. Our results demonstrate that the two phenomena proceed from different mechanisms. Late polykaryocytosis results from an active process dependent on the integrity of the virus genome. In contrast, early polykaryocytosis is a passive process which does not require any de novo virus synthesis but is dependent on the presence of virus proteins in inducer cells.


Vancomycin and teicoplanin use in Victorian hospitals. The Victorian Drug Usage Evaluation Group.
To determine patterns of prescribing of glycopeptide antibiotics (vancomycin and teicoplanin) in Victorian hospitals and identify areas for targeted intervention. A concurrent, observational, multisite evaluation of drug use. Thirty-five Victorian hospitals, 1-14 September 1997. Patients commencing a glycopeptide antibiotic course. Rate of glycopeptide antibiotic use; indications; duration of use; main hospitals using glycopeptide antibiotics. 293 patients (269 adults and 24 neonates) commenced on 302 glycopeptide antibiotic courses: 296 intravenous (i.v.) vancomycin courses and three each of oral vancomycin and parenteral teicoplanin. The overall rate of use was 10.3 courses per 1000 inpatient separations. Of 271 i.v. vancomycin courses for adults, 176 (65%) were for treatment--120 empirically. The median duration of treatment courses was 4.7 days (interquartile range, 2.0-8.2 days). A flucloxacillin-resistant organism was confirmed for 44% of treatment courses. Ninety-five i.v. vancomycin courses were for prophylaxis, including for cardiac (54%) and vascular surgery (21%); 82% of prophylactic courses were administered for less than 24 hours. Of all the glycopeptide antibiotic courses, 69% were administered at five major metropolitan hospitals. Glycopeptide antibiotic use in Victoria is concentrated in the major metropolitan hospitals. Prolonged durations of vancomycin therapy, including for surgical prophylaxis and empirical therapy not subsequently confirmed by microbiology findings, would be suitable targets for interventional strategies.


Preliminary outcomes after transcatheter aortic valve implantation in patients with systemic sclerosis.
The aim of this study was to report the feasibility and the preliminary outcomes after transcatheter aortic valve implantation (TAVI) in the frail and unique high-risk population of patients with systemic sclerosis (SSc) and severe aortic stenosis (AS). All patients with SSc and severe AS treated with TAVI between November 2007 and June 2013 in our centre were included in the current study. Outcomes were assessed using the Valve Academic Research Consortium 2 (VARC-2) definitions. Of 526 surgical high-risk patients treated in the study period, only six had SSc (1.1%). All SSc patients underwent transfemoral TAVI under conscious sedation. The procedure was successfully performed in all patients. There was one case of a major vascular complication. During a median follow-up time of 550 days (interquartile range 390-1,009 days), five of the six patients were alive, free from any complications and with an NYHA Class I or II. One patient died at 670 days from a stroke of unknown aetiology. The present case series suggests that the TAVI procedure is feasible in patients with SSc and severe AS.


The effect of lipoic acid on macro and trace metal levels in living tissues exposed to oxidative stress.
Environmental pollution resulting from fast-paced industrialization, various chemicals used in agriculture, additives in food, smoking and use of alcohol, radiation, some viruses and poor dietary habits all have currently increased the incidence and types of cancer. Polycyclic hydrocarbons are an example of this type of carcinogens. Living things are exposed to this free radical-increasing substance due to various reasons. Oxidative stress caused by reactive oxygen species has an important place in the etiology of cancer, which develops in relation to many factors. Injury caused by cancer in the organism may affect other organs, as well as the tumors organs and tissues. In addition, it is known that some changes take place in the content of macro and trace elements due to cancer in the organism. Our study is intended to explore the protective role of alpha-lipoic acid, which has antioxidant characteristics in living tissues exposed to oxidative stress, in the macro and trace element levels.


Factor X Friuli coagulation disorder. Report of a newly recognized patient and some considerations on the frequency of the disease in the Friuli area.
A newly recognized patient with the factor X Friuli coagulation disorder is presented. The propositus is a 10-year-old boy who presented excessive bleeding after a tooth extraction. This is the 11th patient with this peculiar coagulation disorder discovered in Friuli and the 12th so far described. His parents are consanguineous and heterozygotes together with other family members. The main laboratory features, typical of the disease were prolonged prothrombin time, a prolonged partial thromboplastin time, and a normal Stypven-cephalin clotting time. The incidence of the homozygotes in the Friuli area appear to be 0.000005 (11 cases in a population of 2,000,000). The probable frequency of the abnormal f gene is 0.0033, whereas that of the normal gene F is 0.9967. The theoretical frequency of the heterozygote, Ff condition is 0.007, namely 7 heterozygotes every 1,000 persons. However, only about 70 ascertained heterozygotes have been discovered so far.


Allee effects within small populations of Aconitum napellus ssp. lusitanicum, a protected subspecies in northern France.
Plants growing at low density can suffer from Allee effects as a result of pollen limitation. Previous studies of Allee effects have focused on the effects of variation among populations in size or density on reproduction. Here, the effects of plant distribution within populations on fitness components are explored in a rare plant, Aconitum napellus ssp. lusitanicum, and ecological and genetic mechanisms underlying these effects are identified. To detect pollen limitation, seed production was compared under natural versus hand-supplemented pollinations on inflorescences of different sizes in natural patches differing both in flower density and in isolation from other patches. Germination rate and juvenile survival of seeds produced in low- and high-density patches were also compared. Pollen-supplemented flowers always produced more seeds than open-pollinated flowers, especially among small plants and plants growing at low density. Offspring produced in low-density patches exhibited lower fitness that those produced in high-density patches. This could have been caused by post-fertilization mechanisms, including inbreeding depression or differential maternal resource allocation. These results show that Allee effects on fitness components (ecological and genetic Allee effects) occur within A. napellus populations at different spatial scales. The spatial distribution of plants seems to be a crucial factor affecting reproductive output and fitness.


Anti-osteoporosis Effect of Fisetin against Ovariectomy Induced Osteoporosis in Rats: In silico, in vitro and in vivo Activity.
Osteoporosis is a bone related disease that is characterised by bone loss that further increases the susceptibility to bone fractures and bone frailty due to disturbances in the micro-architecture of bone tissue. Fisetin (flavonoids) exhibited anti-inflammatory and antioxidative stress effects against various diseases. In this protocol, we make an effort to comfort the anti-osteoporosis effect of fisetin against ovariectomy (OVX) induced osteoporosis. A docking study of fisetin and alendronate on the estrogen (α and β) and vitamin D receptors was carried out. SaOS-2 (osteoblast like human) cells were used for the estimation of cell proliferation. The OVX induced OVX model was used and three doses of fisetin and alendronate was given to rats till 16 weeks. The hormone levels, bone turnover markers and biochemical parameters were estimated. Fisetin was docked into estrogen (α and β) and vitamin D receptors, resulting in stable complexes with lower binding scores. Fisetin significantly (p < 0.001) exhibited the induction of cell proliferation against the SaOS-2 cells. OVX induced osteoporosis rats exhibited a suppression of body weight and uterus index, after the Fisetin treatment. Fisetin treatment significantly (p < 0.001) improved the level of bone mineral content (BMC), bone mineral density (BMD) and biochemical parameters such as energy, maximum load, stiffness, young modules, maximum stress and reduced the level of 1,25(OH) 2 D3 and E 2 . Fisetin treatment significantly (p < 0.001) declined the level of phosphorus (P), calcium (Ca) and boosted the level of VitD. Fisetin treatment significantly (p < 0.001) reduced the malonaldehyde (MDA) level and enhanced the glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) level in the bone, intestine and hepatic tissue. Fisetin treatment suppressed the cytokines, RANKL/OPG ratio, receptor activator of nuclear factor-κB ligand (RANKL) and improved the level of osteoprotegerin (OPG). The findings suggest that fisetin could be a beneficial phytoconstituent for the treatment and prevention of postmenopausal osteoporotic complications.


Chlamydomonas and Arabidopsis. A dynamic duo.
A translational biology perspective on Chlamydomonas and Arabidopsis might be expected to focus on those genes, cellular components, or biological processes found first in Chlamydomonas and subsequently studied in Arabidopsis. There are indeed many such examples. There are also examples that flow from Arabidopsis to Chlamydomonas in terms of initial discovery and subsequent study. However, the differences can also be illuminating. In this brief essay, we make a case for the pairing of Chlamydomonas and Arabidopsis as model organisms that bracket a major subset of photosynthetic eukaryotes, the green plants (Mishler, 2000). By analogy with the yeast-mouse dyad, this green pair has tremendous potential as we enter an era of renewed interest in comparative biology.


Squamous cell carcinoma of the nipple following radiation therapy for ductal carcinoma in situ: a case report.
Radiation-induced nonmelanoma skin cancer was first reported seven years after the discovery of X-rays, but has received relatively little consideration in the literature. Specifically, nonmelanoma skin cancer after conservative surgery and radiation for early stage breast cancer has not been well studied. We report the case of a woman who developed squamous cell carcinoma of the nipple nine years after conservative surgery and radiation for ductal carcinoma in situ of the ipsilateral breast. We also review the relevant literature available to date. A 66-year-old African-American woman presented to the hospital with a non-healing ulcer of the right nipple. Her past medical history was significant for right breast ductal carcinoma in situ for which she had undergone lumpectomy and whole breast radiation therapy nine years previously. Mammography and magnetic resonance imaging studies were negative for recurrent breast cancer. However, the latter demonstrated abnormal enhancement in the nipple-areolar region. An incisional biopsy of the lesion demonstrated invasive squamous cell carcinoma. Subsequently, the patient underwent wide excision of the nipple-areolar complex. Sentinel lymph-node biopsy was offered but our patient declined. She was considered to have local disease and hence no further treatment was recommended. This case represents the first reported occurrence of squamous cell carcinoma of the nipple to follow conservative surgery and radiation for ductal carcinoma in situ of the ipsilateral breast. It is likely that radiation overexposure resulted in a radiation burn and subsequent radiodermatitis, placing it at risk for squamous cell carcinoma. A diagnosis of squamous cell carcinoma should be considered in a patient with a nipple lesion following radiation therapy for breast cancer.


Randomized Controlled Trial of a Primary Care-Based Child Obesity Prevention Intervention on Infant Feeding Practices.
To determine the effects of a child obesity prevention intervention, beginning in pregnancy, on infant feeding practices in low-income Hispanic families. The Starting Early randomized controlled trial enrolled pregnant women at a third trimester visit. Women (n = 533) were randomly allocated to a standard care control group or an intervention group participating in prenatal and postpartum individual nutrition/breastfeeding counseling and subsequent nutrition and parenting support groups coordinated with well-child visits. Outcome measures included infant feeding practices and maternal infant feeding knowledge at infant age 3 months, using questions adapted from the Infant Feeding Practices Study II and an infant 24-hour diet recall. A total of 456 families completed 3-month assessments. The intervention group had higher prevalence of exclusive breastfeeding on the 24-hour diet recall (42.7% vs 33.0%, P = .04) compared with controls. The intervention group reported a higher percentage of breastfeeding vs formula feeding per day (mean [SD] 67.7 [39.3] vs 59.7 [39.7], P = .03) and was less likely to introduce complementary foods and liquids compared with controls (6.3% vs 16.7%, P = .001). The intervention group had higher maternal infant feeding knowledge scores (Cohen d, 0.29, 95% CI .10-.48). The effect of Starting Early on breastfeeding was mediated by maternal infant feeding knowledge (Sobel test 2.86, P = .004). Starting Early led to increased exclusive breastfeeding and reduced complementary foods and liquids in 3-month-old infants. Findings document a feasible and effective infrastructure for promoting breastfeeding in families at high risk for obesity in the context of a comprehensive obesity prevention intervention. ClinicalTrials.gov: NCT01541761.


Density functional theory studies of electron interaction with DNA: can zero eV electrons induce strand breaks?
The discovery of DNA strand breaks induced by low energy secondary electrons sparks a necessity to elucidate the mechanism. Through theoretical studies based on a sugar-phosphate-sugar model that mimics a backbone section of the DNA strand, it is found that bond cleavages at 3' or 5'C-O sites after addition of an electron are possible with a ca. 10 kcal/mol activation barrier. Moreover, the potential energy surfaces show that dissociation at both sites is highly favorable thermodynamically. Although the phosphate group in DNA is not a favored site for electron attachment because of competitive electron transfer to the bases, any electrons which attach to phosphates on first encounter may induce strand breaks even when the electron energy is near zero eV. These findings have profound implication as low energy secondary electrons are abundantly generated in all types of ionization radiation.


HIV Trafficking Between Blood and Semen During Early Untreated HIV Infection.
Understanding the dynamics of HIV across anatomic compartments is important to design effective eradication strategies. In this study, we evaluated viral trafficking between blood and semen during primary HIV infection in 6 antiretroviral-naive men who have sex with men. Deep sequencing data of HIV env were generated from longitudinal blood plasma, peripheral blood mononuclear cells, and seminal plasma samples. The presence or absence of viral compartmentalization was assessed using tree-based Slatkin-Maddison and distance-based Fst methods. Phylogeographic analyses were performed using a discrete Bayesian asymmetric approach of diffusion with Markov jump count estimation to evaluate the gene flow between blood and semen during primary HIV infection. Levels of DNA from human herpesviruses and selected inflammatory cytokines were also measured on genital secretions collected at baseline to evaluate potential correlates of increased viral migration between anatomic compartments. We detected varying degrees of compartmentalization in all 6 individuals evaluated. None of them maintained viral compartmentalization between blood and seminal plasma throughout the analyzed time points. Phylogeographic analyses revealed that the HIV population circulating in blood plasma populated the seminal compartment during the earliest stages of infection. In our limited data set, we found no association between local inflammation or herpesvirus shedding at baseline and viral trafficking between semen and blood. The early spread of virus from blood plasma to genital tract and the complex viral interplay between these compartments suggest that viral eradication efforts will require monitoring viral subpopulations in anatomic sites and viral trafficking during the course of infection.


Investigating site-selection mechanisms of retroviral integration in supercoiled DNA braids.
We theoretically study the integration of short viral DNA in a DNA braid made up by two entwined double-stranded DNA molecules. We show that the statistics of single integration events substantially differ in the straight and buckled, or plectonemic, phase of the braid and are more likely in the latter. We further discover that integration is most likely close to plectoneme tips, where the larger bending energy helps overcome the associated energy barrier and that successive integration events are spatio-temporally correlated, suggesting a potential mechanistic explanation of clustered integration sites in host genomes. The braid geometry we consider provides a novel experimental set-up to quantify integration in a supercoiled substrate in vitro, and to better understand the role of double-stranded DNA topology during this process.


Extreme Insolation: Climatic Variation Shapes the Evolution of Thermal Tolerance at Multiple Scales.
The climatic variability hypothesis (CVH) is a cornerstone of thermal ecology, predicting the evolution of wider organismal thermal tolerance ranges in more thermally variable environments. Thermal tolerance ranges depend on both upper and lower tolerance limits (critical thermal maxima [[Formula: see text]] and critical thermal minima [[Formula: see text]]), which may show different responses to environmental gradients. To delineate the relative effects of mean and extreme temperatures on thermal tolerances, we conducted a within-latitude comparative test of CVH predictions for army ants (Dorylinae) at multiple scales: across elevations, in seasonal versus aseasonal forests, and in subterranean versus surface microhabitats. Consistent with the CVH, thermally buffered subterranean species had narrower thermal tolerance ranges. Both [Formula: see text] and [Formula: see text] decreased with elevation for subterranean species. In contrast, aboveground species (those exposed to insolation) showed a decrease in [Formula: see text] but no change in [Formula: see text] across elevations. Furthermore, greater seasonal temperature variation in dry forests correlated with increased [Formula: see text] but not [Formula: see text]. These patterns suggest that [Formula: see text] and [Formula: see text] respond to different abiotic selective forces: habitat-specific exposure to extreme insolation corresponds to [Formula: see text] differences but not to [Formula: see text] variation. We predict that increasingly frequent heat spikes associated with climate change will have habitat-specific physiological consequences for ectothermic animals. Models predicting climate change impacts should account for species microhabitat uses and within-latitude differences in temperature seasonality.


Identification of TmcN as a pathway-specific positive regulator of tautomycetin biosynthesis in Streptomyces sp. CK4412.
Tautomycetin (TMC) is a novel activated T-cell-specific immunosuppressive compound with a unique structure, containing an ester bond linkage between a terminal cyclic anhydride moiety and a linear polyketide chain bearing an unusual terminal alkene. A 3 kb gene, tmcN, with a deduced product of 1029 amino acid residues, located on the 3'-terminus of an approximately 70 kb contiguous TMC biosynthetic gene cluster, was found to have amino acid sequence homology with bacterial regulatory proteins. In silico database comparisons revealed that TmcN belongs to the large ATP-binding regulators of the LuxR protein family. Gene disruption of tmcN from the Streptomyces sp. CK4412 chromosome resulted in significantly reduced antifungal activity against Aspergillus niger, as well as the absence of TMC. In addition, complementation by an integrative plasmid carrying tmcN restored TMC biosynthesis, strongly suggesting that TmcN is a positive regulator of TMC biosynthesis. Gene expression analysis by RT-PCR of the TMC biosynthetic genes revealed that a TmcN mutant strain exhibited reduced expression levels for most of the biosynthetic genes except for its own tmcN. It is thus suggested that TmcN is a pathway-specific positive regulator that activates transcription of the TMC biosynthetic pathway genes in Streptomyces sp. CK4412.


Exercise-related abdominal complaints in a large cohort of runners: a survey with a particular focus on nutrition.
Abdominal complaints (AC) during exercise are a common problem in runners. Nutrition is known to play a role in exercise-related AC, but information on the role of habitual dietary intake is limited. We assessed the prevalence of AC in a large cohort of runners, and investigated its association with potential risk factors, with a particular focus on nutritional factors in the habitual diet. A total of 1993 runners completed two online questionnaires: a general questionnaire on, among others, running habits and exercise-related AC and a Food Frequency Questionnaire. Runners with and without either upper AC (UAC) or lower AC (LAC) were compared regarding personal characteristics, running characteristics and habitual dietary intake. 1139 runners (57%) reported AC during and/or up to 3 hours after running: 302 runners (15%) reported UAC, 1115 (56%) LAC and 278 (14%) both. In about one-third of runners with AC, these complaints negatively affected their running. Exercise-related AC were positively associated with female gender, younger age and more intense running. Most associations with nutritional factors were observed only for LAC in men, with a higher intake of energy, all macronutrients and grain products in men with LAC. In both men and women, a higher intake of tea and unhealthy choices were associated with AC. Exercise-related AC were quite prevalent, and in about one-third of the cases, AC impacted their running. Being female, having a younger age and running at higher intensity were positively associated with AC. Some aspects of the habitual diet were associated with AC. Most notable were positive associations for intake of fat, tea and unhealthy choices.


Transcriptional Reactivation of the FMR1 Gene. A Possible Approach to the Treatment of the Fragile X Syndrome.
Fragile X syndrome (FXS) is the most common cause of inherited intellectual disability, caused by CGG expansion over 200 repeats (full mutation, FM) at the 5' untranslated region (UTR) of the fragile X mental retardation 1 (FMR1) gene and subsequent DNA methylation of the promoter region, accompanied by additional epigenetic histone modifications that result in a block of transcription and absence of the fragile X mental retardation protein (FMRP). The lack of FMRP, involved in multiple aspects of mRNA metabolism in the brain, is thought to be the direct cause of the FXS phenotype. Restoration of FMR1 transcription and FMRP production can be obtained in vitro by treating FXS lymphoblastoid cell lines with the demethylating agent 5-azadeoxycytidine, demonstrating that DNA methylation is key to FMR1 inactivation. This concept is strengthened by the existence of rare male carriers of a FM, who are unable to methylate the FMR1 promoter. These individuals produce limited amounts of FMRP and are of normal intelligence. Their inability to methylate the FMR1 promoter, whose cause is not yet fully elucidated, rescues them from manifesting the FXS. These observations demonstrate that a therapeutic approach to FXS based on the pharmacological reactivation of the FMR1 gene is conceptually tenable and worthy of being further pursued.


Effects of vibrotactile feedback and grasp interface compliance on perception and control of a sensorized myoelectric hand.
Current myoelectric prosthetic limbs are limited in their ability to provide direct sensory feedback to users, which increases attentional demands and reliance on visual cues. Vibrotactile sensory substitution (VSS), which can be used to provide sensory feedback in a non-invasive manner has demonstrated some improvement in myoelectric hand control. In this work, we developed and tested two VSS configurations: one with a single burst-rate modulated actuator and another with a spatially distributed array of five coin tactors. We performed a direct comparative assessment of these two VSS configurations with able-bodied subjects to investigate sensory perception, myoelectric control of grasp force and hand aperture with a prosthesis, and the effects of interface compliance. Six subjects completed a sensory perception experiment under a stimulation only paradigm; sixteen subjects completed experiments to compare VSS performance on perception and graded myoelectric control during grasp force and hand aperture tasks; and ten subjects completed experiments to investigate the effect of mechanical compliance of the myoelectric hand on the ability to control grasp force. Results indicated that sensory perception of vibrotactile feedback was not different for the two VSS configurations in the absence of active myoelectric control, but it was better with feedback from the coin tactor array than with the single actuator during myoelectric control of grasp force. Graded myoelectric control of grasp force and hand aperture was better with feedback from the coin tactor array than with the single actuator, and myoelectric control of grasp force was improved with a compliant grasp interface. Further investigations with VSS should focus on the use of coin tactor arrays by subjects with amputation in real-world settings and on improving control of grasp force by increasing the mechanical compliance of the hand.


An update on current treatment strategies for laryngopharyngeal reflux symptoms.
Laryngopharyngeal reflux (LPR) is a syndrome caused by reflux of gastric contents into the pharynx or larynx, which leads to symptoms of throat clearing, hoarseness, pain, globus sensation, cough, excess mucus production in the throat, and dysphonia. LPR is a challenging condition, as there is currently no gold standard for diagnosis or treatment, and thus this presents a burden to the healthcare system. Strategies for treatment of LPR are numerous. Medical therapies include proton pump inhibitors, which are first line, H2 receptor antagonists, alginates, and baclofen. Other noninvasive treatment options include lifestyle therapy and the external upper esophageal sphincter compression device. Endoscopic and surgical options include antireflux surgery, magnetic sphincter augmentation, and transoral incisionless fundoplication. Functional laryngeal disorders and laryngeal hypersensitivity can present as LPR symptoms with or without gastroesophageal reflux disease. Though there are minimal studies in this area, neuromodulators and behavioral interventions are potential treatment options. Given the complexity of these patients and numerous available treatment options, we propose a treatment algorithm to help clinicians diagnose and triage patients into an appropriate therapy.


The elusive antigen. Thoughts on the structure/function relationship of homogeneous human antibodies.
The rapid progress in the elucidation of antibody structure and function had a major impact on our understanding of monoclonal immunoglobulins. It appears that monoclonicity is a basic immunologic phenomenon, closely related to specificity. The amplification of this phenomenon, observed in pathological conditions such as the various plasma cell dyscrasias, has not been fully understood due to the lack of identifiable antigens capable of triggering this response. The functional and structural data accumulated from animal experiments and the clinical investigation of anti-dextran, anti-phosphorylcholine, anti-thioflavine and homogeneous anti-arsonate antibodies as well as of cold agglutinins, point to the presence of a common structure/function relationship in these molecules. Structural and serologic data, obtained from homogeneous antibodies isolated by us from the serum of patients with cancer of the colon and of the urinary bladder, revealed significant structural similarities expressed by HVR homology and cross-idiotypic specificity.


Suppression of tumorigenicity of glioblastoma cells by adenovirus-mediated MMAC1/PTEN gene transfer.
Mutated in multiple advanced cancers 1/phosphatase and tensin homologue (MMAC1/PTEN) is a novel tumor suppressor gene candidate located on chromosome 10 that is commonly mutated in human glioblastoma multiforme and several other cancer types. To evaluate the function of this gene as a tumor suppressor, we constructed a replication-defective adenovirus (MMCB) for efficient, transient transduction of MMAC1 into tumor cells. Infection of MMAC1-mutated U87MG glioblastoma cells with MMCB resulted in dose-dependent exogenous MMAC1 protein expression as detected by Western blotting of cell lysates. In vitro proliferation of U87MG cells was inhibited by MMCB in comparison to several control adenoviruses at equal viral doses, implying a specific effect of MMAC1 expression. Anchorage-independent growth in soft agar was also inhibited by MMCB compared to control adenovirus. Tumorigenicity in nude mice of transiently transduced mass cell cultures was then assessed. MMCB-infected U87MG cells were almost completely nontumorigenic compared to untreated and several control adenovirus-treated cells at equal viral doses. These data support an in vivo tumor suppression activity of MMAC1/PTEN and suggest that in vivo gene transfer with this recombinant adenoviral vector has a potential use in cancer gene therapy.


Cholinesterase research outreach project (CROP): point of care cholinesterase measurement in an Australian agricultural community.
Australian farmers are routinely exposed to a wide variety of agrichemicals, including herbicides and insecticides. Organophosphate (OP) insecticides are widely used for agricultural production, horticulture and animal husbandry practices. Symptoms of OP toxicity are the results of inhibition of the enzyme acetylcholinesterase (AChE) which is found in many types of conducting tissue in human bodies such as nerve and muscle, central and peripheral tissues, motor and sensory fibres. Cholinesterase can be measured in red blood cells/erythrocytes (AChE) and plasma (PChE). This study aims to explore integration of AChE monitoring into routine health checks for those at risk and also to examine any association between AChE activity and agrichemical use in a Victorian farming community in Australia. This was a prospective cohort study, where farmers and non-famers were compared on the levels of AChE at four time points of baseline, 3-4 weeks, 6-weeks and at 9-weeks. Study participants (N = 55) were residents from South West Victoria, aged between 18 and 75 years, spoke English, and had not had a previous known acute chemical accident. A total of 41 farming (had been farming for more than 5 years) and a convenience sample of 14 non-farming individuals met the inclusion criteria. Testing of AChE was repeated for all participants with a maximum of three times over 10 weeks. The integration of AChE monitoring was very well accepted by all participants. There was no significant difference in average AChE activity between farming and non-farming participants (one-way ANOVA p > 0.05) in this study. There was no significant difference between personal use of agricultural chemicals on farm and the levels of AChE at baseline (measurement 1) or any of the follow up periods (p > 0.05). However, the mean activity of AChE was significantly lower within follow up periods [F (2.633, 139.539) = 14.967, p < 0.001]. There was a significant reduction of AChE between the follow up at 3-weeks and 6-weeks period (p = 0.015). The routine monitoring of AChE may allow for early recognition of chronic low-level exposure to OPs when they are used by farmers, provided a reasonable estimate of baseline AChE is available. This work provides an evidence for recommending the integration of AChE monitoring into point of care (POC) procedures in rural health clinics and quantifying pesticide exposure and personal protection both on the farm and in the home. Farmer engagement is crucial to the successful integration of AChE monitoring into rural health clinics in Australia. ACTRN12613001256763 .


Myelin basic protein and proteolipid protein reactivity of brain- and cerebrospinal fluid-derived T cell clones in multiple sclerosis and postinfectious encephalomyelitis.
T cells were directly cloned from autopsied MS brain plaque tissue and reactivity was measured with the major encephalitogenic neuroantigens, myelin basic protein (MBP), and proteolipid protein (PLP). Control clones were simultaneously derived from the blood. The proportion of T4+ and T8+ T cell clones from the brain tissue differed from that of peripheral blood T cell clones derived at the same time, suggesting that the clones were not derived from the peripheral blood. None of 57 brain-derived T cell clones proliferated to either MBP or PLP, although they responded well to PHA and IL 2. An additional 235 clones derived from the cerebrospinal fluid and 126 clones from the peripheral blood of other subjects with multiple sclerosis also did not proliferate to MBP or PLP. In contrast, five of nine T4+ clones from the CSF of a subject with postinfectious encephalomyelitis exhibited low but clear reactivity to human MBP, supporting the possible role of MBP as the target antigen in this disease. These studies, the first to clone T cells directly from MS plaque tissue, suggest that the lack of consistent T cell reactivity to MBP or PLP in the peripheral blood of MS patients does not appear to be secondary to the sequestration of a large number of these cells in the brain.


The impact of preterm birth on hospital inpatient admissions and costs during the first 5 years of life.
To compare the cumulative use and cost of hospital inpatient services to 5 years of age by individuals divided into 4 subgroups by gestational age at birth. Costs applied to the hospital service utilization profile of each infant born in 2 areas covered by the Oxford Record Linkage Study during 1970-1993. Oxfordshire and West Berkshire, southern United Kingdom. 239 694 individuals divided into 4 subgroups by gestational age at birth: <28 weeks, 28 to 31 weeks, 32 to 36 weeks, >or=37 weeks. Number and duration of hospital admissions during the first 5 years of life and costs, expressed in pound sterling and valued at 1998-1999 prices, of hospital inpatient services. The total duration of hospital admissions for infants born at <28 and at 28 to 31 gestational weeks was 85 and 16 times that for term infants, respectively, once duration of life had been taken into account. Hospital inpatient service costs were significantly higher for preterm infants than for term infants, with the cost differences persisting throughout infancy and early and mid-childhood. Over the first 5 years of life, the adjusted mean cost difference was estimated at pound 14,614 ( 22,798 US dollars) when infants born at <28 weeks gestational age were compared with term infants and pound 11,958 ( 18,654 US dollars) when infants born at 28 to 31 weeks gestational age were compared with term infants. Independent contributions to total cost came from being born: small for gestational age, a multiple, during the 1970s and early 1980s, to a woman of extreme maternal age or who was hospitalized antenatally, and from experiencing extended survival or childhood disease. However, preterm birth remained the strongest predictor of high cost. Preterm birth is a major predictor of how much an individual will cost hospital service providers during the first 5 years of life.


Independent regulation of adherens and tight junctions by tyrosine phosphorylation in Caco-2 cells.
To study the role of tyrosine phosphorylation in the control of intercellular adhesion of intestinal cells, we have generated several clones of Caco-2 cells that express high levels of pp60v-src only after addition of butyrate. Expression of this oncogene in cells 5 days after confluence induced beta-catenin and p120-ctn tyrosine phosphorylation, redistribution of E-cadherin to the cytosol and disassembly of adherens junctions. However, tight junctions of Caco-2 cells at 5 days after confluence were not altered by expression of pp60v-src. Similar results were obtained when Caco-2 cells were incubated with phosphotyrosine phosphatase inhibitor orthovanadate. Although addition of this compound to postconfluent cells disrupt adherens junctions, tight junctions remain unaltered, as determined measuring monolayer permeability to mannitol or hyperphosphorylation of Triton-insoluble occludin. Modifications in tight junction permeability of Caco-2 were only observed at high concentrations of orthovanadate (1 mM). Interestingly, this tyrosine phosphorylation-refractory state was achieved after confluence since early postconfluent cells (day 2) showed a limited but significant response to low doses of orthovanadate. These results suggest that tight junctions of differentiated Caco-2 cells are uncoupled from adherens junctions and are insensitive to regulation by tyrosine phosphorylation.


Investigation of the genotoxic effects of patent blue V (E131) in human peripheral lymphocytes and in silico molecular docking.
Patent Blue V (PBV) is a water-soluble synthetic dyestuff that is used as a coloring agent in the food industry and for medical imaging in health monitoring. The aim of this study was to investigate the in vitro clastogenic, aneugenic and cytotoxic effects of PBV in human peripheral lymphocytes using micronucleus assay, comet assay, as well as plasmid DNA interaction and bacterial AMES tests. In addition to in vitro tests, the affinity of PBV against DNA was determined by molecular docking analysis in silico. PBV produced significant MN formation only at high doses and longer treatment time, however, it did not significantly affect the nuclear division index (NDI). Furthermore, PBV was unable to cause DNA single-strand breaks and significant oxidative damage on the pBR322 plasmid DNA and it didn't reverse the harmful effects caused by the clastogenic treatment of UV + H2O2 on plasmid DNA. In the Ames test, no significant increase was detected in the number of revertant colonies of mutant strains, TA98 and TA100, following PBV treatment. No significant molecular interaction between B-DNA and PBV occured in molecular docking simulations. In conclusion, PBV had no significant genotoxic and cytotoxic effects in this study. However, considering that the information intensity related to the genotoxic effects of PBV in the literature is still insufficient, reports of further studies with different genotoxicity endpoints will be needed to elucidate the exact genotoxic feature.


Use of drug-eluting stents as a function of predicted benefit: clinical and economic implications of current practice.
Benefits of drug-eluting stents (DES) in percutaneous coronary intervention (PCI) are greatest in those at the highest risk of target-vessel revascularization (TVR). Drug-eluting stents cost more than bare-metal stents (BMS) and necessitate prolonged dual antiplatelet therapy (DAPT), which increases costs, bleeding risk, and risk of complications if DAPT is prematurely discontinued. Our objective was to assess whether DES are preferentially used in patients with higher predicted TVR risk and to estimate if lower use of DES in low-TVR-risk patients would be more cost-effective than the existing DES use pattern. We analyzed more than 1.5 million PCI procedures in the National Cardiovascular Data Registry (NCDR) CathPCI registry from 2004 through 2010 and estimated 1-year TVR risk with BMS using a validated model. We examined the association between TVR risk and DES use and the cost-effectiveness of lower DES use in low-TVR-risk patients (50% less DES use among patients with <10% TVR risk) compared with existing DES use. There was marked variation in physicians' use of DES (range 2%-100%). Use of DES was high across all predicted TVR risk categories (73.9% in TVR risk <10%; 78.0% in TVR risk 10%-20%; and 83.2% in TVR risk >20%), with a modest relationship between TVR risk and DES use (relative risk, 1.005 per 1% increase in TVR risk [95% CI, 1.005-1.006]). Reducing DES use by 50% in low-TVR-risk patients was projected to lower US health care costs by $205 million per year while increasing the overall TVR event rate by 0.5% (95% CI, 0.49%-0.51%) in absolute terms. Use of DES in the United States varies widely among physicians, with only a modest correlation to patients' risk of restenosis. Less DES use among patients with low risk of restenosis has the potential for significant cost savings for the US health care system while minimally increasing restenosis events.


Common oligosaccharide moieties inhibit the adherence of typical and atypical respiratory pathogens.
Intervention in bacterial adhesion to host cells is a novel method of overcoming current problems associated with antibiotic resistance. Antibiotic-resistant strains of bacteria that cause respiratory tract infections are a problem in hospitals and could be used in bioterrorist attacks. A range of bacterial species was demonstrated to attach to an alveolar epithelial (A549) cell line. In all cases, cell surface oligosaccharides were important in attachment, demonstrated by reduced adhesion when A549 cells were pre-treated with tunicamycin. Bacillus anthracis and Yersinia pestis displayed a restricted tropism for oligosaccharides compared to the environmental, opportunistic pathogens, Pseudomonas aeruginosa, Burkholderia cenocepacia, Burkholderia pseudomallei and Legionella pneumophila. The compound with the greatest anti-adhesion activity was p-nitrophenol. Other generic attachment inhibitors included the polymeric saccharides (dextran and heparin), GalNAcbeta1-4Gal, GalNAcbeta1-3Gal, Galbeta1-4GlcNAc and Galbeta1-3GlcNAc. Burkholderia pseudomallei attachment was particularly susceptible to oligosaccharide inhibition. Combinations of such compounds may serve as a novel generic therapeutics for respiratory tract infections.


Use of blood components in critically ill patients in the medical intensive care unit of a tertiary care hospital.
The art of fluid administration and hemodynamic support is one of the most challenging aspects of treating critically ill patients. Transfusions of blood products continue to be an important technique for resuscitating patients in the intensive care settings. Concerns about the rate of inappropriate transfusion exist, particularly given the recognized risks of transfusions and the decreasing availability of donor blood. We investigated the current transfusion practice in the critically ill patients at our hospital. A total of 1817 consecutive critically ill patients admitted between January 2006 and December 2006 were included in this retrospective study. The blood request forms of the patients were analyzed, and their pretransfusion investigations, indications for transfusions, etc. were studied. Nine hundred and eleven (50.1%) critically ill patients, comprising 71.6% males and 28.4% females, received blood/blood components. About 43.8% patients were administered packed red cells (PRC), 18.27% fresh frozen plasma (FFP) and 8.4% transfused platelets. Among those receiving PRC, 31.1% had a pretransfusion Hb below 7.5g%, 34.4% had Hb between 7.5 and 9g%, while 21.4% had Hb above 9g%. Among those receiving FFP, 14.5% had an international normalized ratio INR < 1.5, and 19% had a pretransfusion platelet count above 50,000/cumm. During the study, there were 7% of the patients who received red cells and FFP, 2% of the patients received red cells and platelets, 1% of the patients received platelets and FFP, and 5% of the patients had received all the three components, i.e., red cells, FFP and Platelets. The baseline investigations and/or clinical indications were not mentioned in 13.1% of patients receiving PRC, 57% receiving FFP and 49.7% receiving platelets. About 21.4% of PRC, 14.5% of FFP, and 19% of platelets were inappropriately indicated. Clinicians in our centre were conservative in keeping with recent transfusion guidelines. A significant number of blood request forms were still incomplete with baseline investigations not mentioned in the request forms.


Beyond ManifoldEM: geometric relationships between manifold embeddings of a continuum of 3D molecular structures and their 2D projections.
ManifoldEM is an established method of geometric machine learning developed to extract information on conformational motions of molecules from their projections obtained by cryogenic electron microscopy (cryo-EM). In a previous work, in-depth analysis of the properties of manifolds obtained for simulated ground-truth data from molecules exhibiting domain motions has led to improvements of this method, as demonstrated in selected applications of single-particle cryo-EM. In the present work this analysis has been extended to investigate the properties of manifolds constructed by embedding data from synthetic models represented by atomic coordinates in motion, or three-dimensional density maps from biophysical experiments other than single-particle cryo-EM, with extensions to cryo-electron tomography and single-particle imaging with a X-ray free-electron laser. Our theoretical analysis revealed interesting relationships between all these manifolds, which can be exploited in future work.


Children's oral health in the medical curriculum: a collaborative intervention at a university-affiliated hospital.
The purpose of this study was to 1) describe the structure of the oral health program in a university-affiliated hospital; 2) evaluate staff's knowledge and attitudes toward oral health; and 3) propose ways to strengthen the incorporation of oral health prevention for children into clinical medical education. Qualitative methods were used to evaluate the program. Structured interviews with seventeen medical center personnel were conducted, and clinic utilization reports provided ICD-9 diagnostic frequency and visits. Clinic staff, pediatric residents, dental and pediatric faculty, hospital administrators, and clinic directors were interviewed. The themes identified during these interviews were motivation, roles, operational and organizational issues, and integration into the larger medical care system. Integration of an early childhood caries prevention program into the clinical medical education curriculum can be accomplished. After implementation of the oral health program described in this paper, dental caries became the eleventh most common diagnosis seen in the clinic when previously it did not appear in the top forty. However, institutional and organizational barriers are significant. Barriers identified were 1) lack of clarity in defining leadership and roles regarding oral health, 2) time and work overload in a busy pediatric clinic, 3) a tracking system was not available to quickly determine which children needed caries prevention procedures and education, and 4) billing and medical record form changes could not be fully established prior to starting the program.


Survey of the year 2009: applications of isothermal titration calorimetry.
Isothermal titration calorimetry (ITC) is now an established and invaluable method for determining the thermodynamic constants, association constant and stoichiometry of molecular interactions in aqueous solutions. The technique has become widely used by biochemists to study protein interaction with other proteins, small molecules, metal ions, lipids, nucleic acids and carbohydrates; and nucleic acid interaction with small molecules. The drug discovery industry has utilized this approach to measure protein (or nucleic acid) interaction with drug candidates. ITC has been used to screen candidates, guide the design of potential drugs and validate the modelling used in structure-based drug design. Emerging disciplines including nanotechnology and drug delivery could benefit greatly from ITC in enhancing their understanding and control of nano-particle assembly, and drug binding and controlled release.


LcrV of Yersinia pestis enters infected eukaryotic cells by a virulence plasmid-independent mechanism.
Yersinia pestis is the causative agent of bubonic plague and possesses a set of plasmid-encoded, secretable virulence proteins termed LcrV and Yops which are essential for survival in mammalian hosts. Yops and LcrV are secreted by a type III mechanism (Ysc), and Yops are unidirectionally targeted into the cytosol of associated eukaryotic cells in a tissue culture infection model. LcrV is required for Yops targeting, and recent findings have revealed that it can localize to the bacterial surface; however, its fate in this infection model has not been investigated in detail. In this study, we compared the localization of LcrV to that of the targeted proteins YopE and YopM by immunoblot analysis of fractions of Yersinia-infected HeLa cultures or by laser-scanning confocal microscopy of infected monolayers. Both LcrV and YopE were secreted by contact-activated, extracellularly localized yersiniae and were targeted to the HeLa cell cytosol. Although a significant amount of LcrV partitioned to the culture medium (unlike YopE), this extracellular pool of LcrV was not the source of the LcrV that entered HeLa cells. Unlike targeting of YopE and YopM, targeting of LcrV occurred in the absence of a functional Ysc apparatus and other virulence plasmid (pCD1)-expressed proteins. However, the Ysc is necessary for LcrV to be released into the medium, and our recent work has shown that localization of LcrV on the bacterial surface requires the Ysc. These results indicate that two mechanisms exist for the secretion of LcrV by Y. pestis, both of which are activated by contact with eukaryotic cells. LcrV secreted by the Ysc reaches the bacterial surface and the surrounding medium, whereas the second is a novel, Ysc-independent pathway which results in localization of LcrV in the cytosol of infected cells but not the surrounding medium.


Various enzyme activities in muscle and other organs of dystrophic mice.
To elucidate the metabolic abnormality of musclar dystrophy, 27 kinds of enzyme activity in various organs of control and dystrophic mice were examined. The organs examined included muscle, bone, heart, testis, uterus, spleen, thymus, submaxillary gland, stomach, pancreas, liver, kidney, brain, and lung. The activities of 14 different aminopeptidases, 5 endopeptidases, 4 glycosidases, phosphatase, esterase, and ribonuclease were measured. Most of the enzyme activities were significantly elevated in muscles and bones of dystrophic mice. These organs were similar in their patterns of enzyme abnormality. Among the 14 kinds of aminopeptidase activity studied, the degree of increased activity was greater for the aminopeptidases (AP):Ala-AP, Leu-AP, Met-AP, Phe-AP, Trp-AP, Gly-Pro-Leu-AP. In addition to aminopeptidases, there were significant increases in activities of chymotrypsinlike enzyme, cathepsin C, cathepsin D, several glycosidases and neutral ribonuclease in the muscles of dystrophic mice. Similarly increased enzyme activity was also observed in organs other than muscle and bone. Furthermore, protein content in most organs was higher in dystrophic mice than in those of control mice. These abnormalities were seen in both males and females. The present results suggest that there are extensive abnormalities in the protein metabolism in dystrophic mice. It seems therefore that the therapeutic approach to muscular dystrophy should be studies not only from the well-known abnormality of intramuscular endopeptidases, but from other aspects as well.