Systematic review of vision-related quality of life questionnaires for older institutionalised seniors with dementia.
In view of our interest in helping older institutionalised adults with co-existing visual and cognitive deficits, our objective was to review vision-related quality of life (VRQoL) questionnaires developed for people with dementia. A systematic literature search was performed in seven relevant databases, limited to peer-reviewed journals published in English, French, Spanish, Italian or German languages, between 1982 and 2012. The retained VRQoL questionnaires were described. Only relevant articles that fully meet our target population and interest criteria were eligible for psychometric properties rating. The literature search identified 461 potentially relevant articles, from which 69 were isolated and reviewed. Eight articles presented questionnaires that were administered to older institutionalised individuals with dementia. Three VRQoL questionnaires were identified, from which only one was designed and validated for those with mild to moderate dementia and none for individuals with more advanced dementia. There is a pressing need to build and validate a questionnaire for assessing VRQoL in older institutionalised individuals with dementia, to provide a research tool capable of evaluating how changes in vision impact their QoL.


cGMP Signaling, Phosphodiesterases and Major Depressive Disorder.
DEFICITS IN NEUROPLASTICITY ARE HYPOTHESIZED TO UNDERLIE THE PATHOPHYSIOLOGY OF MAJOR DEPRESSIVE DISORDER (MDD): the effectiveness of antidepressants is thought to be related to the normalization of disrupted synaptic transmission and neurogenesis. The cyclic adenosine monophosphate (cAMP) signaling cascade has received considerable attention for its role in neuroplasticity and MDD. However components of a closely related pathway, the cyclic guanosine monophosphate (cGMP) have been studied with much lower intensity, even though this signaling transduction cascade is also expressed in the brain and the activity of this pathway has been implicated in learning and memory processes. Cyclic GMP acts as a second messenger; it amplifies signals received at postsynaptic receptors and activates downstream effector molecules resulting in gene expression changes and neuronal responses. Phosphodiesterase (PDE) enzymes degrade cGMP into 5'GMP and therefore they are involved in the regulation of intracellular levels of cGMP. Here we review a growing body of evidence suggesting that the cGMP signaling cascade warrants further investigation for its involvement in MDD and antidepressant action.


Trypanosomes and microfilariae in feral owl and squirrel monkeys maintained in research colonies.
A group of 358 owl and squirrel monkeys imported from Colombia, Peru, and Bolivia for the U.S. Agency for International Development Malaria Vaccine Development Program was examined for trypanosomes and microfilariae. Trypanosoma rangeli, isolated by hemoculture from Aotus nancymai, Saimiri b. boliviensis, and S. b. peruviensis, accounted for 76.6% of all trypanosome infections. Trypanosoma cruzi was isolated from 25 of 194 S. b. boliviensis, including two mixed infections with T. rangeli. Identifications of trypanosomes were confirmed by blinded tests with a panel of five rRNA probes on a subsample of cultures identified morphologically. Although no trypanosomes were isolated from Aotus vociferans or A. lemurinus griseimembra, positive serologic responses to T. cruzi were observed by indirect immunofluorescence assay in all species of monkeys examined and ranged from 42.1% among S. b. peruviensis to 92.3% among A. vociferans. Among T. rangeli-infected monkeys, 43.7% were seronegative for T. cruzi. No microfilariae were found in S. b. boliviensis or A. l. griseimembra. Mansonella barbascalensis and Dipetalonema caudispina were observed in A. vociferans, M. panamensis in A. nancymai, and M. saimiri and D. caudispina in S. b. peruviensis. Such naturally occurring infections in imported animal models are potential sources of accidental transmission to animal handlers and uninfected laboratory animals and can introduce confounding variables into otherwise well-planned and well-executed studies.


Nascent peptides that block protein synthesis in bacteria.
Although the ribosome is a very general catalyst, it cannot synthesize all protein sequences equally well. For example, ribosomes stall on the secretion monitor (SecM) leader peptide to regulate expression of a downstream gene. Using a genetic selection in Escherichia coli, we identified additional nascent peptide motifs that stall ribosomes. Kinetic studies show that some nascent peptides dramatically inhibit rates of peptide release by release factors. We find that residues upstream of the minimal stalling motif can either enhance or suppress this effect. In other stalling motifs, peptidyl transfer to certain aminoacyl-tRNAs is inhibited. In particular, three consecutive Pro codons pose a challenge for elongating ribosomes. The translation factor elongation factor P, which alleviates pausing at polyproline sequences, has little or no effect on other stalling peptides. The motifs that we identified are underrepresented in bacterial proteomes and show evidence of stalling on endogenous E. coli proteins.


Comparison of instream methods for measuring hydraulic conductivity in sandy streambeds.
Streambed hydraulic conductivity (K) values were determined at seven stream transects in the Platte River Basin in Nebraska using different instream measurement techniques. Values were compared to determine the most appropriate technique(s) for use in sandy streambeds. Values of K determined from field falling- and constant-head permeameter tests analyzed using the Darcy equation decreased as permeameter diameter increased. Seepage meters coupled with hydraulic gradient measurements failed to yield K values in 40% of the trials. Consequently, Darcy permeameter and seepage meter tests were not preferred approaches. In the upper 0.25 m of the streambed, field falling- and constant-head permeameter tests analyzed with the Hvorslev solution generally had similar K values that were significantly greater than those determined using the Hazen grain-size, Bouwer and Rice slug test for anisotropic and isotropic conditions, and Alyamani and Sen grain-size methods; median differences between these tests and the Hvorslev falling-head 60 cm diameter permeameter were about 8, 9, 17, and 35 m/day, respectively. The Hvorslev falling-head permeameter test is considered the most robust method for measuring K of the upper 0.25 m of the streambed because of the inherent limitations of the empirical grain-size methods and less sediment disturbance for permeameter than slug tests. However, lateral variability in K along transects on the Platte, North Platte, and Wood Rivers was greater than variability in K between valid permeameter, grain-size, or slug tests, indicating that the method used may matter less than making enough measurements to characterize spatial variability adequately. At the Platte River tributary sites, the upper 0.3 m of the streambed typically had greater K than sediment located 0.3 to 2.5 m below the streambed surface, indicating that deposits below the streambed may limit ground water/surface water fluxes. The Hvorslev permeameter tests are not a practical measurement approach for these greater depths. Thus, selection of a method for measuring streambed K needs to consider the vertical location of the sediments that are most likely to limit the rate of ground water/surface water interaction.


A peptide-stainless steel reaction that yields a new bioorganic-metal state of matter.
A synthetic peptide derived from the native protein sequence of a metal binding bacterial pilus was observed to spontaneously react with stainless steel via a previously unreported type of chemical interaction to generate an altered form of stainless steel which we term bioorganic stainless steel. Bioorganic stainless steel has a significantly increased electron work function (4.9 ± 0.05 eV compared to 4.79 ± 0.07 eV), decreased material adhesive force (19.4 ± 8.8 nN compared to 56.7 ± 10.5 nN), and is significantly harder than regular 304 stainless steel (~40% harder). A formal or semi-formal organo-metallic covalent bond is generated between a pilin receptor binding domain and stainless steel based on XPS analysis which indicates that the electronic state of the surface is altered. Further, we establish that the peptide-steel reaction demonstrates a degree of stereospecificity as the reaction of native L-peptide, D-peptide and a retro-inverso-D-peptide yields bioorganic steel products that can be differentiated via the resulting EWF (4.867 ± 0.008 eV, 4.651 ± 0.008 eV, and 4.919 ± 0.007 eV, respectively). We conclude that electron sharing between the peptide and steel surface results in the stabilization of surface electrons to generate bioorganic steel that displays altered properties relative to the initial starting material. The bioorganic steel generated from the retro-inverso-D-peptide yields a protease stable product that is harder (41% harder at a 400 μN load), and has a 50% lower corrosion rate compared with regular stainless steel (0.11 ± 0.03 mpy and 0.22 ± 0.04 mpy, respectively). Bioorganic steel is readily fabricated.


Exosomes and Their Role in Cancer Progression.
Exosomes are a subset of extracellular vesicles and their size is approximately 100 nm in diameter. They are surrounded by a lipid bilayer membrane and secreted from almost all of cells. Exosomes are generated within the endocytic system as ILV (intraluminal membrane vesicle) and secreted during the fusion of MVB (multivesicular body) with the cell membrane. Recently it has been reported that exosomes modulate cell-cell communication contributing to the maintenance of tissue homeostasis by molecules including exosomes. Moreover, exosomes released from cancer cells are involved in cancer progression. Thus, data regarding the role of the exosomes in malignant cancer will lead to development of novel diagnostic and therapeutic methods.


Post-Stroke Lingual Dystonia: Clinical Description and Neuroimaging Findings.
Lingual dystonia is extremely rare following stroke. We describe clinical features and neuroimaging findings in a series of 11 patients (seven acute and four chronic) with post-stroke lingual dystonia and review the literature. This was a case series using a preformed structured proforma and review of literature using a PubMed search. In our case series, all patients had dysarthria as a presenting symptom. Seven patients had acute presentation (six had an ischemic infarct and one had thalamic hemorrhage) and four had chronic presentation (all had infarct). All patients except one had small infarcts, with the majority of them in the basal ganglia and subcortical white matter regions. Additional chronic ischemic lesions were seen in all patients with acute presentation. The majority of the patients with acute (five out of seven; 71.42%) presentation had left-sided involvement on imaging. We could identify only one case of acute post-stroke lingual dystonia following the PubMed search. Three other cases of post-stroke lingual dystonia with chronic presentation have been described; however, these were associated with oromandibular or cranial dystonia. Our results, based on brain lesions, suggest that all lingual dystonia patients with acute infarcts had underlying chronic infarcts. Overall, more left-sided than right-sided strokes were observed with post-stroke lingual movement disorders including dystonia; however, the data were not significant (p = 1). All patients had dysarthria, with only one having mild tongue weakness and only four having facial weakness. This suggests that the lingual dystonia was responsible for the dysarthria rather than weakness in these patients.


Understanding deregulated cellular and molecular dynamics in the haematopoietic stem cell niche to develop novel therapeutics for bone marrow fibrosis.
Bone marrow fibrosis is the continuous replacement of blood-forming cells in the bone marrow with excessive scar tissue, leading to failure of the body to produce blood cells and ultimately to death. Myofibroblasts are fibrosis-driving cells and are well characterized in solid organ fibrosis, but their role and cellular origin in bone marrow fibrosis have remained obscure. Recent work has demonstrated that Gli1+ and leptin receptor+ mesenchymal stromal cells are progenitors of fibrosis-causing myofibroblasts in the bone marrow. Genetic ablation or pharmacological inhibition of Gli1+ mesenchymal stromal cells ameliorated fibrosis in mouse models of myelofibrosis. Conditional deletion of the platelet-derived growth factor (PDGF) receptor-α (PDGFRA) gene (Pdgfra) and inhibition of PDGFRA by imatinib in leptin receptor+ stromal cells suppressed their expansion and ameliorated bone marrow fibrosis. Understanding the cellular and molecular mechanisms in the haematopoietic stem cell niche that govern the mesenchymal stromal cell-to-myofibroblast transition and myofibroblast expansion will be critical to understand the pathogenesis of bone marrow fibrosis in both malignant and non-malignant conditions, and will guide the development of novel therapeutics. In this review, we summarize recent discoveries of mesenchymal stromal cells as part of the haematopoietic niche and as myofibroblast precursors, and discuss potential therapeutic strategies in the specific targeting of fibrotic transformation in bone marrow fibrosis. © 2018 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.


The severity of nonalcoholic fatty liver disease is associated with gut dysbiosis and shift in the metabolic function of the gut microbiota.
Several animal studies have emphasized the role of gut microbiota in nonalcoholic fatty liver disease (NAFLD). However, data about gut dysbiosis in human NAFLD remain scarce in the literature, especially studies including the whole spectrum of NAFLD lesions. We aimed to evaluate the association between gut dysbiosis and severe NAFLD lesions, that is, nonalcoholic steatohepatitis (NASH) and fibrosis, in a well-characterized population of adult NAFLD. Fifty-seven patients with biopsy-proven NAFLD were enrolled. Taxonomic composition of gut microbiota was determined using 16S ribosomal RNA gene sequencing of stool samples. Thirty patients had F0/F1 fibrosis stage at liver biopsy (10 with NASH), and 27 patients had significant F≥2 fibrosis (25 with NASH). Bacteroides abundance was significantly increased in NASH and F≥2 patients, whereas Prevotella abundance was decreased. Ruminococcus abundance was significantly higher in F≥2 patients. By multivariate analysis, Bacteroides abundance was independently associated with NASH and Ruminococcus with F≥2 fibrosis. Stratification according to the abundance of these two bacteria generated three patient subgroups with increasing severity of NAFLD lesions. Based on imputed metagenomic profiles, Kyoto Encyclopedia of Genes and Genomes pathways significantly related to NASH and fibrosis F≥2 were mostly related to carbohydrate, lipid, and amino acid metabolism. NAFLD severity associates with gut dysbiosis and a shift in metabolic function of the gut microbiota. We identified Bacteroides as independently associated with NASH and Ruminococcus with significant fibrosis. Thus, gut microbiota analysis adds information to classical predictors of NAFLD severity and suggests novel metabolic targets for pre-/probiotics therapies.


Fibre intake in relation to serum total cholesterol levels and CHD risk: a comparison of dietary assessment methods.
Prospective diet diaries may be more accurate than retrospective food frequency questionnaires (FFQ). The objective of this study was to compare FFQ and 7-day diet diary (7DD) measurements of fibre intake with the incidence of coronary heart disease (CHD). We compared 7DD and FFQ fibre intake in a nested case-control study in a population of 25639 men and women aged 40-79 years, surveyed in 1993-97 and followed up until 2007. Among 2151 CHD cases and 5354 controls, FFQ and 7DD fibre intake (6 g/day) was examined in relation to serum total cholesterol and CHD using linear and logistic regression adjusted for age and additionally for body mass index, physical activity, smoking status, family history of CHD, social class, diabetes, alcohol, energy, saturated fat and use of lipid lowering medication, antihypertensive medication or aspirin. Age-adjusted serum total cholesterol was inversely associated with 7DD fibre among men and women, but with FFQ fibre among men only. In the multivariate analysis, associations with 7DD fibre were attenuated among men (regression coefficient -0.036 mmol/l, s.e. 0.021, P-value 0.087) and women (regression coefficient -0.069 mmol/l, s.e. 0.036, P-value 0.053), and were non-significant for FFQ fibre. Among men, age-adjusted CHD risk was inversely associated with 7DD fibre (odds ratio (OR) 0.84, 95% confidence interval (CI) 0.79-0.90), but not with FFQ fibre (OR 0.96, 95% CI 0.90-1.12). Among women, age-adjusted CHD risk was inversely associated with 7DD fibre (OR 0.83, 95% CI 0.75-0.93), and had a weaker inverse borderline-significant association with FFQ fibre (OR 0.93, 95% CI 0.87-1.01). Multivariate models yielded similar results. Inconsistencies in diet-CHD relationships in population studies may be associated with the use of different dietary assessment methods.


Human motor control of landing from a drop in simulated microgravity.
Landing on the ground on one's feet implies that the energy gained during the fall be dissipated. The aim of this study is to assess human motor control of landing in different conditions of fall initiation, simulated gravity, and sensory neural input. Six participants performed drop landings using a trapdoor system and landings from self-initiated counter-movement jumps in microgravity conditions simulated in a weightlessness environment by different pull-down forces of 1-, 0.6-, 0.4-, and 0.2 g External forces applied to the body, orientation of the lower limb segments, and muscular activity of 6 lower limb muscles were recorded synchronously. Our results show that 1) subjects are able to land and stabilize in all experimental conditions; 2) prelanding muscular activity is always present, emphasizing the capacity of the central nervous system to approximate the instant of touchdown; 3) the kinetics and muscular activity are adjusted to the amount of energy gained during the fall; 4) the control of landing seems less finely controlled in drop landings as suggested by higher impact forces and loading rates, plus lower mechanical work done during landing for a given amount of energy to be dissipated. In conclusion, humans seem able to adapt the control of landing according to the amount of energy to be dissipated in an environment where sensory information is altered, even under conditions of non-self-initiated falls.


Environmental persistence of amphibian and reptilian ranaviruses.
Ranaviruses infect fish, amphibians, and reptiles. The present study was conducted to compare the persistence of amphibian and reptilian ranaviruses in a pond habitat. The 4 viruses used in this study included 2 amphibian ranaviruses, Frog virus 3 (FV3, the type species of the genus Ranavirus) and an isolate from a frog, and 2 ranaviruses of reptilian origin (from a tortoise and from a gecko). A sandwich germ-carrier technique was used to study the persistence of these viruses in sterile and unsterile pond water (PW) and soil obtained from the bank of a pond. For each virus, virus-loaded carriers were placed in each of the 3 substrates, incubated at 4 and 20°C, and titrated at regular intervals. Serial data were analyzed using a linear regression model to calculate T-90 values (time required for 90% reduction in the virus titer). Resistance of the viruses to drying was also studied. All 4 viruses were resistant to drying. At 20°C, T-90 values of the viruses were 22 to 31 d in sterile PW and 22 to 34 d in unsterile PW. Inactivation of all 4 viruses in soil at this temperature appeared to be non-linear. T-90 values at 4°C were 102 to 182 d in sterile PW, 58 to 72 d in unsterile PW, and 30 to 48 d in soil. Viral persistence was highest in the sterile PW, followed by the unsterile PW, and was lowest in soil. There were no significant differences in the survival times between the amphibian and reptilian viruses. The results of the present study suggest that ranaviruses can survive for long periods of time in pond habitats at low temperatures.


Mutations defective in ribonucleotide reductase activity interfere with pollen plastid DNA degradation mediated by DPD1 exonuclease.
Organellar DNAs in mitochondria and plastids are present in multiple copies and make up a substantial proportion of total cellular DNA despite their limited genetic capacity. We recently demonstrated that organellar DNA degradation occurs during pollen maturation, mediated by the Mg(2+) -dependent organelle exonuclease DPD1. To further understand organellar DNA degradation, we characterized a distinct mutant (dpd2). In contrast to the dpd1 mutant, which retains both plastid and mitochondrial DNAs, dpd2 showed specific accumulation of plastid DNAs. Multiple abnormalities in vegetative and reproductive tissues of dpd2 were also detected. DPD2 encodes the large subunit of ribonucleotide reductase, an enzyme that functions at the rate-limiting step of de novo nucleotide biosynthesis. We demonstrated that the defects in ribonucleotide reductase indirectly compromise the activity of DPD1 nuclease in plastids, thus supporting a different regulation of organellar DNA degradation in pollen. Several lines of evidence provided here reinforce our previous conclusion that the DPD1 exonuclease plays a central role in organellar DNA degradation, functioning in DNA salvage rather than maternal inheritance during pollen development.


3,4-dimethoxyphenyl bis-benzimidazole, a novel DNA topoisomerase inhibitor that preferentially targets Escherichia coli topoisomerase I.
Antibiotic resistance in bacterial pathogens is a serious clinical problem. Novel targets are needed to combat increasing drug resistance in Escherichia coli. Our objective is to demonstrate that 2-(3,4-dimethoxyphenyl)-5-[5-(4-methylpiperazin-1-yl)-1H-benzimidazol-2yl]-1H-benzimidazole (DMA) inhibits E. coli DNA topoisomerase I more strongly than human topoisomerase I. In addition, DMA is non-toxic to mammalian cells at antibiotic dosage level. In the present study, we have established DMA as an antibacterial compound by determining MICs, post-antibiotic effects (PAEs) and MBCs for different standard as well as clinical strains of E. coli. We have described the differential catalytic inhibitory mechanism of bis-benzimidazole, DMA, for human and E. coli topoisomerase I and topoisomerase II by performing different assays, including relaxation assays, cleavage-religation assays, DNA unwinding assays, ethidium bromide displacement assays, decatenation assays and DNA gyrase supercoiling assays. DMA significantly inhibited bacterial growth at a very low concentration, but did not affect human cell viability at higher concentrations. Activity assays showed that it preferentially targeted E. coli topoisomerase I over human topoisomerase I, topoisomerase II and gyrase. Cleavage-religation assays confirmed DMA as a poison inhibitor of E. coli topoisomerase I. This study illuminates new properties of DMA, which may be further modified to develop an efficient topoisomerase inhibitor that is selective towards bacterial topoisomerase I. This is the first report of a bis-benzimidazole acting as an E. coli topoisomerase I inhibitor. DMA is a safe, non-cytotoxic molecule to human cells at concentrations that are needed for antibacterial activity.


Endothelium-targeted overexpression of heat shock protein 27 ameliorates blood-brain barrier disruption after ischemic brain injury.
The damage borne by the endothelial cells (ECs) forming the blood-brain barrier (BBB) during ischemic stroke and other neurological conditions disrupts the structure and function of the neurovascular unit and contributes to poor patient outcomes. We recently reported that structural aberrations in brain microvascular ECs-namely, uncontrolled actin polymerization and subsequent disassembly of junctional proteins, are a possible cause of the early onset BBB breach that arises within 30-60 min of reperfusion after transient focal ischemia. Here, we investigated the role of heat shock protein 27 (HSP27) as a direct inhibitor of actin polymerization and protectant against BBB disruption after ischemia/reperfusion (I/R). Using in vivo and in vitro models, we found that targeted overexpression of HSP27 specifically within ECs-but not within neurons-ameliorated BBB impairment 1-24 h after I/R. Mechanistically, HSP27 suppressed I/R-induced aberrant actin polymerization, stress fiber formation, and junctional protein translocation in brain microvascular ECs, independent of its protective actions against cell death. By preserving BBB integrity after I/R, EC-targeted HSP27 overexpression attenuated the infiltration of potentially destructive neutrophils and macrophages into brain parenchyma, thereby improving long-term stroke outcome. Notably, early poststroke administration of HSP27 attached to a cell-penetrating transduction domain (TAT-HSP27) rapidly elevated HSP27 levels in brain microvessels and ameliorated I/R-induced BBB disruption and subsequent neurological deficits. Thus, the present study demonstrates that HSP27 can function at the EC level to preserve BBB integrity after I/R brain injury. HSP27 may be a therapeutic agent for ischemic stroke and other neurological conditions involving BBB breakdown.


Development of Kaempferol-Loaded Gelatin Nanoparticles for the Treatment of Corneal Neovascularization in Mice.
Cornea is the transparent layer in front of the eye that does not contain blood vessels. Among eye diseases, corneal neovascularization (NV) is one of the major causes of vision loss, since it can also lead to blindness. An herbal extraction containing flavonoid, kaempferol (KA), with antiangiogenic effect was chosen as a candidate drug for inhibited vessel formation. The use of nanomedicine has led to higher drug bioavailability and slow release of the drug as an effective therapeutic formulation in ocular drug delivery. In this study, we prepared gelatin nanoparticles (GNP) with kaempferol encapsulation (GNP-KA) for corneal NV treatment by topical delivery, i.e., eye drops. We found that GNP with/without KA loading was in the size of 85-150 nm, and its zeta potential was around 22-26 mV. The KA entrapment rate of GNP-KA was around 90-98%, and the loading rate was about 4.6%. The TEM results clearly indicated the GNP-KA NPs to be round spheres. The in vitro test involved the adoption of human umbilical vein endothelial cells (HUVECs) for coculture with these nanoparticles. From WST-8 assay, and cell migration examinations, it was evident that GNP-KA had the capacity to inhibit the cell viability and function of HUVECs. The results from in vivo tests such as ocular vessels observation, hematoxylin & eosin (H&E) stain, and metalloproteinases (MMP)/vascular endothelial growth factor (VEGF) quantification revealed the mice's eyes with corneal NV treated by eye drops containing GNP-KA once daily for 7 days had better therapeutic effects with less vessels in-growths in the cornea, compared to the KA solution group by reducing the production of MMP and VEGF in the cornea. Therefore, we expected to achieve a comfortable treatment with a simple method using nanomedicine (GNP-KA) as ophthalmological agent delivered as eye drops.


Alcohol in decomposed bodies: postmortem synthesis and distribution.
Blood alcohol (ethanol) concentrations in decomposed bodies can mean drinking during life and/or endogenous production after death. The correct interpretation is important in medicolegal cases. This retrospective study of 286 autopsied medical examiner cases was undertaken to evaluate alcohol concentrations and distribution in various fluids and tissues in decomposed bodies. Cases with alcohol present were classified as endogenous production, ingestion, or unable to determine based upon one or more of the following criteria: the presence of ethanol in only one of more than one body fluids, an atypical distribution of ethanol in body fluids, reliable scene or historical information, the presence of C3 alcohols in body fluids. Alcohol was classified as endogenously produced in 55 cases. The presence of alcohol was attributed to ingestion in 130 cases. No alcohol was detected in 39 cases. We were unable to determine the source of the remaining 62 alcohol concentrations. The highest blood alcohol concentration derived from endogenous production was 0.07% in the cases with other fluids negative. The mean blood alcohol concentration was 0.06% and ranged as high as 0.16% in cases having atypical ratios. Alcohol was found in blood and bile while urine and vitreous fluid were negative or had lower concentrations in cases with endogenous production. We conclude that for the majority of cases in which endogenous blood production of alcohol occurs the concentration in blood may be as high as 0.15%.


Systems-level analysis of proteolytic events in increased vascular permeability and complement activation in skin inflammation.
During inflammation, vascular permeability is increased by various proteolytic events, such as the generation of bradykinin, that augment local tissue responses by enabling tissue penetration of serum proteins, including complement and acute-phase proteins. Proteases also govern inflammatory responses by processing extracellular matrix proteins and soluble bioactive mediators. We quantified changes in the proteome and the nature of protein amino termini (the N-terminome) and the altered abundance of murine proteases and inhibitors during skin inflammation. Through analysis of the N-terminome by iTRAQ-TAILS, we identified cotranslational and posttranslational αN-acetylation motifs, quantitative increases in protein abundance, and qualitative changes in the proteolytic signature during inflammation. Of the proteins identified in normal skin, about half were cleaved, and phorbol ester-induced inflammation increased the proportion of cleaved proteins, including chemokines and complement proteins, that were processed at previously uncharacterized sites. In response to phorbol ester-induced inflammation, mice deficient in matrix metalloproteinase 2 (MMP2) showed reduced accumulation of serum proteins in the skin and exhibited different proteolytic networks from those of wild-type mice. We found that the complement 1 (C1) inhibitor attenuated the increase in serum protein accumulation in inflamed skin. Cleavage and inactivation of the C1 inhibitor by MMP2 increased complement activation and bradykinin generation in wild-type mice, leading to increased vessel permeability during inflammation, which was diminished in Mmp2(-/-) mice. Thus, our systems-level analysis of proteolysis dissected cleavage events associated with skin inflammation and demonstrated that loss of a single protease could perturb the proteolytic signaling network and enhance inflammation.


Feasibility study of a take-home array-based functional electrical stimulation system with automated setup for current functional electrical stimulation users with foot-drop.
To investigate the feasibility of unsupervised community use of an array-based automated setup functional electrical stimulator for current foot-drop functional electrical stimulation (FES) users. Feasibility study. Gait laboratory and community use. Participants (N=7) with diagnosis of unilateral foot-drop of central neurologic origin (>6mo) who were regular users of a foot-drop FES system (>3mo). Array-based automated setup FES system for foot-drop (ShefStim). Logged usage, logged automated setup times for the array-based automated setup FES system and diary recording of problems experienced, all collected in the community environment. Walking speed, ankle angles at initial contact, foot clearance during swing, and the Quebec User Evaluation of Satisfaction with Assistive Technology version 2.0 (QUEST version 2.0) questionnaire, all collected in the gait laboratory. All participants were able to use the array-based automated setup FES system. Total setup time took longer than participants' own FES systems, and automated setup time was longer than in a previous study of a similar system. Some problems were experienced, but overall, participants were as satisfied with this system as their own FES system. The increase in walking speed (N=7) relative to no stimulation was comparable between both systems, and appropriate ankle angles at initial contact (N=7) and foot clearance during swing (n=5) were greater with the array-based automated setup FES system. This study demonstrates that an array-based automated setup FES system for foot-drop can be successfully used unsupervised. Despite setup's taking longer and some problems, users are satisfied with the system and it would appear as effective, if not better, at addressing the foot-drop impairment. Further product development of this unique system, followed by a larger-scale and longer-term study, is required before firm conclusions about its efficacy can be reached.


Prediction of low-back trouble frequency in a working population.
This study was performed to estimate the discriminatory power of multiple combinations of risk indicators for the occurrence and recurrence of low-back trouble (LBT) in workers. Two categories of LBT provided groups for discrimination; 1) the presence or absence of LBT history, and 2) three patterns of recurrence characterized by the number of episodes (isolated, periodic, chronic). The risk indicators comprised data reflecting occupational and leisure demands on the back, measures of lumbar sagittal mobility, and anamnestic features of the first episode. Discriminant analysis was the statistical procedure used. The results showed that it was possible to find linear combinations of the discriminating variables that successfully allocated around two-thirds of the sample to the correct group. The presence of a history of LBT was predicted by the combined effect of increasing age and adult sports participation, but only in females did a heavier job contribute to such prediction. A reduction in risk was associated with lumbar flexibility and sports participation at school. Chronic LBT was more accurately identified than the two other groups; increasing age, a long initial spell, and an onset early in life were associated with increased likelihood of chronicity, while a report of symptoms being relieved by sitting reduced this risk. It is concluded that the occurrence and recurrence of LBT are related to combinations of risk indicators, and that it is imperative to consider the interactive effect of a multiplicity of factors in epidemiologic studies.


Evidence that vesicles containing living, virulent Mycobacterium tuberculosis or Mycobacterium avium in cultured human macrophages are not acidic.
Mycobacterium tuberculosis and Mycobacterium avium multiply in cultured human macrophages (MP) within membrane-enclosed vesicles. These vesicles are generally assumed to be acidic. The evidence most frequently cited for this assumption is that pyrazinamide, which requires an acid pH to be effective, is effective and streptomycin, which loses most of its activity at a low pH, is poorly effective against tubercle bacilli. This assumption was tested by using the two weak bases chloroquine and NH4Cl to raise the pH of acidic vesicles in MP experimentally infected with M. tuberculosis or M. avium. An immunocytochemical locator of acidic regions in the MP was used to monitor the association of intracellular bacilli with acidity. MP were infected with M. tuberculosis or M. avium and incubated with various combinations of the drugs and the weak bases. Replication of the bacteria in the MP was measured by culture counts. Intracellular associations of the mycobacteria with acidity were assessed by electron micrographs and by using the weak base 3-(2,4-dinitroanilino)-3'-amino-N-methyl dipropylamine, which was detected with colloidal gold-labeled antibodies. It was confirmed by immunocytochemistry that both chloroquine and NH4Cl raise the pH of acidic vesicles in the infected MP. However, neither caused any pH-related change in the antimycobacterial activities of pyrazinamide or streptomycin or of the pH-independent drug isoniazid. Immunochemical analyses showed acidity to be associated with killed but not living mycobacteria in the MP. These findings suggest that living M. tuberculosis and M. avium are located in human MP in vesicles which are not acidic.


The ROCK inhibitor eye drop accelerates corneal endothelium wound healing.
To evaluate the effect of Rho kinase (ROCK)-inhibitor eye drops on a corneal endothelial dysfunction primate model and human clinical case series of corneal endothelial dysfunction. As a corneal-endothelial partially injured model, the corneal endothelium of seven cynomolgus monkeys was damaged by transcorneal freezing; 10 mm of rock inhibitor Y-27632 was then applied topically 6 times daily. The phenotype of the reconstructed corneal endothelium was evaluated by immunohistochemical analysis and noncontact specular microscopy. For clinical study, the effect of Y-27632 eye drops after transcorneal freezing was evaluated in eight corneal endothelial dysfunction patients: four central corneal edema patients and four diffuse corneal edema patients. Slit-lamp microscopy revealed that both Y-27632-treated and -nontreated corneas became hazy after transcorneal freezing, and then recovered their transparency within 4 weeks. ROCK inhibitor Y-27632 promoted recovery of corneal endothelial cell density and wound healing in terms of both morphology and function. The percentage of ZO-1 and Na(+)/K(+)-ATPase positive cells in the regenerated area in the Y-27632 group was significantly higher than in the controls. Noncontact specular microscopy revealed that corneal endothelial cell density was significantly higher in the Y-27632 group compared with the controls at 4 weeks; cell density reached approximately 3000 cells/mm(2), as opposed to 1500 cells/mm(2) in the control group. In addition to the animal study findings, the clinical study findings showed that Y-27632 eye drops effectively improved corneal edema of corneal endothelial dysfunction patients with central edema. These findings show that rock inhibitor Y-27632 eye drops promote corneal endothelial wound healing in a primate animal model and suggest the possibility of Y-27632 as a novel therapeutic modality for certain forms of corneal endothelial dysfunction. (http://www.umin.ac.jp/ctr/ number, UMIN000003625.).


Comparison of anxiety, depression, and post-traumatic stress symptoms in relatives of ICU patients in an American and an Indian public hospital.
An intensive care unit (ICU) admission of a patient causes considerable stress among relatives. Whether this impact differs among populations with differing sociocultural factors is unknown. The aim was to compare the psychological impact of an ICU admission on relatives of patients in an American and Indian public hospital. A cross-sectional study was carried out in ICUs of two tertiary care hospitals, one each in major metropolitan cities in the USA and India. A total of 90 relatives visiting patients were verbally administered a questionnaire between 48 hours and 72 hours of ICU admission that included the Hospital Anxiety and Depression Scale (HADS), Beck Depression Inventory-II (BDI-II) and Impact of Events Scale-Revised (IES-R) for post-traumatic stress response. Statistical analysis was done using the Mann-Whitney and chi-square tests. Relatives in the Indian ICU had more anxiety symptoms (median HADS-A score 11 [inter-quartile range 9-13] vs. 4 [1.5-6] in the American cohort; P<0.0001), more depression symptoms (BDI-II score 14 [8.5-19] vs. 6 [1.5-10.5], P<0.0001) but a comparable post-traumatic stress response (IES-R score>30). 55% of all relatives had an incongruous perception regarding "change in the patient's condition" compared to the objective change in severity of illness. "Change in worry" was incongruous compared to the perception of improvement of the patient's condition in 78% of relatives. Relatives of patients in the Indian ICU had greater anxiety and depression symptoms compared to those in the American cohort, and had significant differences in factors that may be associated with this psychological impact. Both groups showed substantial discordance between the perceived and objective change in severity of illness.


Lim homeobox genes in the Ctenophore Mnemiopsis leidyi: the evolution of neural cell type specification.
Nervous systems are thought to be important to the evolutionary success and diversification of metazoans, yet little is known about the origin of simple nervous systems at the base of the animal tree. Recent data suggest that ctenophores, a group of macroscopic pelagic marine invertebrates, are the most ancient group of animals that possess a definitive nervous system consisting of a distributed nerve net and an apical statocyst. This study reports on details of the evolution of the neural cell type specifying transcription factor family of LIM homeobox containing genes (Lhx), which have highly conserved functions in neural specification in bilaterian animals. Using next generation sequencing, the first draft of the genome of the ctenophore Mnemiopsis leidyi has been generated. The Lhx genes in all animals are represented by seven subfamilies (Lhx1/5, Lhx3/4, Lmx, Islet, Lhx2/9, Lhx6/8, and LMO) of which four were found to be represented in the ctenophore lineage (Lhx1/5, Lhx3/4, Lmx, and Islet). Interestingly, the ctenophore Lhx gene complement is more similar to the sponge complement (sponges do not possess neurons) than to either the cnidarian-bilaterian or placozoan Lhx complements. Using whole mount in situ hybridization, the Lhx gene expression patterns were examined and found to be expressed around the blastopore and in cells that give rise to the apical organ and putative neural sensory cells. This research gives us a first look at neural cell type specification in the ctenophore M. leidyi. Within M. leidyi, Lhx genes are expressed in overlapping domains within proposed neural cellular and sensory cell territories. These data suggest that Lhx genes likely played a conserved role in the patterning of sensory cells in the ancestor of sponges and ctenophores, and may provide a link to the expression of Lhx orthologs in sponge larval photoreceptive cells. Lhx genes were later co-opted into patterning more diversified complements of neural and non-neural cell types in later evolving animals.


A pilot evaluation of alternating preoperative chemotherapy in the management of patients with locoregionally advanced breast carcinoma.
This prospective trial was conducted to evaluate the outcome of patients treated with preoperative and post operative chemotherapy, mastectomy, and irradiation for locoregionally advanced breast carcinoma. Between June 1986 and September 1990, 71 patients received 2 cycles of doxorubicin that alternated with 2 cycles of cyclophosphamide, methotrexate, and 5-fluorouracil prior to mastectomy; irradiation was administered when the tumor was not amenable to surgical resection. Additional chemotherapy and tamoxifen, in hormone receptor-positive tumors, was used after mastectomy. Post-operative irradiation was given on a selective basis for patients at high risk for locoregional disease recurrence. Although 5 patients (7%) had disease progression, clinical partial or complete tumor response to preoperative chemotherapy was noted in 46 patients (65%). Sixty-eight patients (96%) underwent mastectomy. With a median follow-up of 52 months, the relapse-free and overall survival rates at 5 years were 42% and 57% respectively. Locoregional tumor recurrence occurred in 14 patients (20%), and 28 patients (39%) developed metastatic disease. Menopausal status, clinical presentation (noninflammatory vs. inflammatory), and American Joint Committee on Cancer clinical stage were independent covariates associated with patient outcome. Preoperative alternating chemotherapy, with the selective use of irradiation, resulted in significant locoregional disease regression and the successful integration of mastectomy into the therapeutic strategy. Locoregional tumor control and relapse-free and overall survival estimates for the approach described herein compared favorably with other comtemporary reports for this condition.


Association of raisin consumption with nutrient intake, diet quality, and health risk factors in US adults: National Health and Nutrition Examination Survey 2001-2012.
Raisins are one of the most commonly consumed dried fruits. Because of their unique nutrient profile, raisins may have some distinctive health benefits. The purpose of this study was to examine the cross-sectional association between raisin consumption and nutrient intake, dietary quality, body weight, and metabolic syndrome risk factors in adults. Data from the National Health and Nutrition Examination Survey (NHANES), 2001-2012 (n = 29,684) were used. Raisin consumers (n = 458, 60% female) were defined as those having any amount of raisins during the first 24 h dietary recall. Diet quality was calculated using the Healthy Eating Index-2010 (HEI-2010). Covariate (demographic and lifestyle)-adjusted regression analyses were conducted using appropriate sample weights and significance was set at p < 0.01. Raisin consumers had higher intakes of energy (9%); higher intakes of 'nutrients of public health concern/shortfall nutrients', such as dietary fiber (34%), potassium (16%), magnesium (22%), vitamin C (24%), and vitamin E (22%); and lower intakes of 'nutrients to limit', such as added sugar (-17%), saturated fat (-15%), and sodium (-10%), than non-consumers. No associations were observed for intakes of calcium, iron, vitamin A, vitamin D, and folate. Consumers had higher intakes of total fruit (72%), whole fruit (111%), vegetables (22%), and whole grains (109%), and had a higher diet quality, as indicated by 25% higher total HEI-2010 scores than non-consumers. Compared to non-consumers, raisin consumers had a lower body weight (-4.2%), body mass index (-5.2%), and waist circumference (-3.8%), were 39% less likely to be overweight or obese, and had a 54% reduced risk of metabolic syndrome. In conclusion, raisin consumption was associated with better nutrient intake, diet quality, and weight parameters, and with lower risk of being obese and having metabolic syndrome in US adults.


Isolation and Quantification of Sphingosine and Sphinganine from Rat Serum Revealed Gender Differences.
Sphingolipids are an important group of lipids that play crucial roles in living cells, facilitating cell recognition, signal transduction and endocytosis. The concentration of sphingosine and some of its derivatives like sphinganine may serve as a biomarker for the diagnosis of sphingolipidoses or be used for further research into similar diseases. In this study, a sphingolipid extraction and a high resolution detection method specific for sphingosine and sphinganine was adapted and tested. Lipids were extracted from rats' serum, coupled to o-phthalaldehyde and detected with a fluorescence detector after running through a silica gel column in a high performance liquid chromatography system. With this method, we analysed 20 male and 20 female rat serum samples and compared the concentrations of sphingosine and sphinganine. The results showed a significant difference between the sphingosine concentrations in the male and female rats. The sphingosine concentration in female rats was 805 ng/mL (standard deviation, SD ± 549), while that in males was significantly lower at (75 ng/mL (SD ± 40)). Furthermore, the sphingosine:sphinganine ratio was almost 15-fold higher in the females' samples. The method presented here facilitates the accurate quantification of sphingosine and sphinganine concentrations down to 2.6 ng and 3.0 ng, respectively, and their ratio in small amounts of rat serum samples to study the sphingolipid metabolism and its potential modulation due to gene mutations or the effect of prevalent toxins.


Molecular genetics of herpes simplex virus. II. Mapping of the major viral glycoproteins and of the genetic loci specifying the social behavior of infected cells.
We have mapped the location in herpes simplex virus (HSV) DNA of (i) three mutations at different loci (syn loci) which alter the social behavior of infected cells from clumping of rounded cells to polykaryocytosis, (ii) a mutation which determines the accumulation of one major glycoprotein [VP8.0(C(2))], and (iii) the sequences encoding four major virus glycoproteins [VP8.0(C(2)), VP7(B(2)), VP8.5(A), and VP19E(D(2))]. The experimental design and results were as follows. (i) Analysis of HSV-1 x HSV-2 recombinants showed that the sequences encoding the VP19E(D(2)) glycoprotein map in the S component, whereas the sequences encoding the other three major glycoproteins are in two locations in the L component of HSV DNA. The templates specifying the HSV-1 and HSV-2 glycoprotein VP8.0(C(2)) appear not to be colinear; we isolated recombinants specifying glycoproteins comigrating in sodium dodecyl sulfate-polyacrylamide gels with VP8.0(C(2)) of both HSV-1 and HSV-2. (ii) Marker rescue of a ts mutant defective in accumulation of glycoprotein VP7(B(2)) showed that the mutation maps within a region containing the sequences encoding that glycoprotein. (iii) Marker transfer experiments involving transfection of rabbit skin cells with donor HSV-1(F) DNA and fragments from several donor strains causing fusion of Vero or both Vero and HEp-2 cells revealed the existence of three syn loci specifying the social behavior of cells and one locus (Cr) determining the accumulation of glycoprotein VP8.0(C(2)). The Cr locus maps to the right of the template specifying VP8.0(C(2)) glycoprotein. Loci syn 1 and syn 2 map at or near the Cr locus but can be segregated from it. Locus syn 3 maps at or near the template specifying glycoproteins VP7(B(2)) and VP8.5(A). The expression of mutations in the syn 1 and syn 3 loci appear to be cell type dependent, in that recombinants with these mutations fuse Vero cells but not HEp-2 cells. Recipients of the syn 2 locus or of both syn 2 and syn 1 loci fuse both Vero and HEp-2 cells.


Human nasal rhinosporidiosis: an Italian case report.
Rhinosporidiosis is a disease affecting primarily the mucosa of nose, conjunctiva and urethra. It is endemic in some Asiatic regions, affecting people of any age and sex. Its manifestation is a polypoid mass growing inside the affected cavity and the only treatment is surgical excision. Rhinosporidium seeberi is the aetiological agent. Many discussions arouse regarding the taxonomic classification of the microorganism, recent studies established it is an aquatic protistan parasite. The lesion may recur and sometimes cause osteolytic bone lesions. In endemic areas it is not easy to establish if recurrent lesions are due to relapse or reinfection. A 26-year-old male patient from India, resident in Italy since 2005, presented in March 2006 with a history of nasal obstruction of three months duration. Physical examination showed an erythematous, papillomatous mass, 3 cm in diameter, obstructing the right nasal cavity. A microscopic diagnosis of rhinosporidiosis was made. Few Italian human cases of this disease have been previously reported in the literature. Rhinosporidiosis is a condition which both clinicians and pathologists should keep in mind when managing patients from endemic countries with nasal masses. Moreover, it is very interesting in such cases to follow the clinical course: an eventual recurrence of the lesion in our patient would mean a true relapse, excluding the possibility of a reinfection, more probable in the endemic areas.


Cell type- and subcellular position-dependent summation of unitary postsynaptic potentials in neocortical neurons.
Theoretical studies predict that the modes of integration of coincident inputs depend on their location and timing. To test these models experimentally, we simultaneously recorded from three neocortical neurons in vitro and investigated the effect of the subcellular position of two convergent inputs on the response summation in the common postsynaptic cell. When scattered over the somatodendritic surface, combination of two coincident excitatory or inhibitory synaptic potentials summed linearly in layer 2/3 pyramidal cells, as well as in GABAergic interneurons. Slightly sublinear summation with connection specific kinetics was observed when convergent inputs targeted closely placed sites on the postsynaptic cell. The degree of linearity of summation also depended on the type of connection, the relative timing of inputs, and the activation state of I(h). The results suggest that, when few inputs are active, the majority of afferent permutations undergo linear integration, maintaining the importance of individual inputs. However, compartment- and connection-specific nonlinear interactions between synapses located close to each other could increase the computational power of individual neurons in a cell type-specific manner.


Adolescents' Exposure to Online Risks: Gender Disparities and Vulnerabilities Related to Online Behaviors.
In the last decade, readily available electronic devices have created unprecedented opportunities for teens to access a wide variety of information and media-both positive and negative-on the internet. Despite the increasing number of initiatives taking place worldwide intended to assess and mitigate the online risks encountered by children and adolescents, there is still a need for a better understanding of how adolescents use the internet and their susceptibility to exposure to risks in the online space. We conducted a cross-sectional online survey of a convenience sample of 733 8th and 9th grade students in Utah. The survey contained eight questions regarding students' exposure to three types of online risk scenarios: content risk, contact risk, and criminal risk. Independent variables included students' online behaviors, use of social media and private messaging apps, and adult supervision of online activities. Logistic and negative binomial regression models indicated that gender, social media use, and chatting with strangers were associated with exposure to multiple risky online scenarios. Our results provide critical information to educators involved in the development of initiatives focusing on the reduction of youth online risk by identifying correlates of risky online events, allowing them to tailor their initiatives to meet the needs of potentially vulnerable populations.


Performance monitoring in monkey frontal eye field.
The frontal eye fields (FEF) are thought to mediate response selection during oculomotor decision tasks. In addition, many FEF neurons have robust postsaccadic responses, but their role in postchoice evaluative processes (online performance monitoring) is only beginning to become apparent. Here we report error-related neural activity in FEF while monkeys performed a biased speed-categorization task that enticed the animals to make impulsive errors. Twenty-three percent of cells in macaque FEF coded an internally generated error-related signal, and many of the same cells also coded task difficulty. The observed responses are primarily consistent with three related concepts that have been associated with performance monitoring: (1) response conflict; (2) uncertainty; and (3) reward prediction. Overall, our findings suggest a novel role for the FEF as part of the neural network that evaluates the preceding choice to optimize behavior in the future.


E-cadherin gene mutations provide clues to diffuse type gastric carcinomas.
The calcium-dependent homophilic cell adhesion molecule and candidate suppressor gene, E (epithelial)-cadherin, plays a major role in the organization and integrity of most epithelial tissues. Diffusely growing gastric carcinomas show markedly reduced homophilic cell-to-cell interactions. We speculated that mutations in the E-cadherin gene may be responsible for the scattered phenotype of this type of carcinoma. For that reason we have examined E-cadherin in 26 diffuse type, 20 intestinal type and 7 mixed gastric carcinomas (Laurén's classification) at the DNA, RNA, and protein levels. Reverse transcription polymerase chain reaction and direct sequencing of amplified E-cadherin complementary DNA fragments revealed inframe skipping of either exon 8 or exon 9 in 10 patients with diffuse tumors and an exon 9 deletion in one patient with a mixed carcinoma; both exons encode putative calcium binding domains. These alterations were not seen in nontumorous gastric tissues. Splice site mutations responsible for the exon deletions were identified in six of these patients, eliminating the possibility of alternative splicing mechanisms. Five of these splice site alterations were confirmed as somatic mutations. Non-splice site mutations were observed in three diffuse type tumors, namely a 69-base pair deletion of exon 10 and two point mutations, one of which destroys a putative calcium binding region. Immunohistochemical evaluation showed E-cadherin immunoreactivity in tumors and lymph node metastases of patients expressing abnormal mRNA. The allelic status of the E-cadherin gene was analyzed in one patient, revealing loss of heterozygosity with retention of a mutated E-cadherin allele. Overall, E-cadherin mutations were identified in 50% (13 of 26) of the diffuse type and in 14% (1 of 7) of the mixed carcinomas. In contrast, two silent E-cadherin mutations (not changing the amino acid sequence) were detected in two tumors of the intestinal type. Our study provides strong in vivo evidence that E-cadherin gene mutations may contribute to the development of diffusely growing gastric carcinomas and support a tumor/metastasis suppressor gene hypothesis.


A cross-sectional analysis of the cost and affordability of achieving recommended intakes of non-starchy fruits and vegetables in the capital of Vanuatu.
The low-income Pacific Island nation of Vanuatu is experiencing a double burden of diet-related disease whereby micronutrient deficiencies and underweight occur at the same time as obesity related non-communicable diseases. Increasing intakes of nutrient dense, energy dilute foods such as fruits and vegetables will be important to address this issue. However, reduced access to agricultural land in urban areas provides limited opportunities for traditional subsistence fruit and vegetable production. Set in Port Vila, Vanuatu's capital and main urban centre, this study aimed to determine the cost and affordability of meeting international recommendations to consume at least 400 g of non-starchy fruits and vegetables (NSFV) per person per day, and assess the adequacy of households' NSFV expenditure. NSFV prices from the 2010 Vanuatu Consumer Price Index (n = 56) were used to determine the minimum monthly cost of purchasing 400 g of local NSFV per person, after accounting for wastage. The 2010 Vanuatu Household Income and Expenditure Survey (n = 578 households) was analysed to determine the proportion of households' total and food budget required to purchase 400 g of local NSFV for all household members. Household NSFV costs were also compared against actual household expenditure on these items. Consumption of own-produce and gifts received were included within estimates of food expenditure. The minimum cost of purchasing the recommended amount of local NSFV was 1,486.24 vatu ($16.60 US) per person per month. This level of expenditure would require an average of 9.6% (SD 6.4%) of households' total budget and 26.3% (SD 25.8%) of their food budget. The poorest households would need to allocate 40.9% (SD 34.3%) of their total food budget to NSFV to purchase recommended amounts of these foods. Twenty-one percent of households recorded sufficient NSFV expenditure while 23.4% recorded less than 10% of the expenditure required to meet the NSFV recommendations. Achieving recommended intakes of local NSFV in Port Vila is largely unaffordable, and expenditure on these foods was inadequate for most households in Port Vila in 2010. Addressing fruit and vegetable affordability will be an important consideration in prevention of non-communicable diseases in the Pacific region.


Metabolism of androsta-1,4,6-triene-3,17-dione and detection by gas chromatography/mass spectrometry in doping control.
The urinary metabolism of the irreversible aromatase inhibitor androsta-1,4,6-triene-3,17-dione was investigated. It is mainly excreted unchanged and as its 17beta-hydroxy analogue. For confirmation, 17beta-hydroxyandrosta-1,4,6-trien-3-one was synthesized and characterized by nuclear magnetic resonance (NMR) in addition to the parent compound. In addition, several reduced metabolites were detected in the post-administration urines, namely 17beta-hydroxyandrosta-1,4-dien-3-one (boldenone), 17beta-hydroxy-5beta-androst-1-en-3-one (boldenone metabolite), 17beta-hydroxyandrosta-4,6-dien-3-one, and androsta-4,6-diene-3,17-dione. The identification was performed by comparison of the metabolites with reference material utilizing gas chromatography/mass spectrometry (GC/MS) of the underivatized compounds and GC/MS and GC/tandem mass spectrometry (MS/MS) of their trimethylsilyl (TMS) derivatives. Alterations in the steroid profile were also observed, most obviously in the androsterone/testosterone ratio. Even if not explicitly listed, androsta-1,4,6-triene-3,17-dione is classified as a prohibited substance in sports by the World Anti-Doping Agency (WADA) due to its aromatase-inhibiting properties. In 2006 three samples from human routine sports doping control tested positive for metabolites of androsta-1,4,6-triene-3,17-dione. The samples were initially found suspicious for the boldenone metabolite 17beta-hydroxy-5beta-androst-1-en-3-one. Since metabolites of androst-4-ene-3,6,17-trione were also present in the urine samples, it is presumed that these findings were due to the administration of a product like 'Novedex Xtreme', which could be easily obtained from the sport supplement market.


Partnering with persons in long-term recovery from substance use disorder: experiences from a collaborative research project.
Traditional research about substance use disorder (SUD) treatment is considered, among an increasing number of service users, to be disempowering and poorly reflective of their priorities. Thus, this methodological article sought to examine the experiences of a peer research group (PRG), whose four members were in long-term SUD recovery, and a principal investigator (PI), when collaborating on a study of SUD recovery. This article has also aspired to discern the influence of peer researcher participation on the research process. The purpose of the qualitative research project that formed the basis of this methodological study was to examine the reasons provided and strategies employed for abstaining from problematic substance use among persons with SUDs. The project took place from 2015 to 2018, during which time individual interviews were conducted with 18 persons in recovery from SUDs. The PRG contributed to all parts of the project and worked alongside the PI in preparing the study, during early stages of data analysis, and while writing up the findings. In total, ten group discussions were held over the course of 3 years. The study showed that the PRG offered important contributions with respect to developing the interview guide, preunderstanding among the PRG members, and discussing alternative forms of data collection. Key findings about how this collaborative research process was experienced relate to three matters: the group aspect of participation, the value of predictable routines and clear expectations, and the open sharing of private matters. The PI experienced the research process as having been enriched by alternative ways of asking questions and interpreting findings and as an interactive arena for reciprocal social and professional support. When establishing a PRG while studying recovery processes, it can be advantageous to include several peer researchers with diverse lived experiences concerning substance use, treatment, and recovery. If possible, at least one peer researcher with formal training or qualitative research experience might be included. The PI should be trained in collaborating with peer researchers or should be part of a research environment in which it is possible to discuss methodological challenges with other researchers.


Cationic lipid-mediated gene transfer: current concepts.
Infectious disease, heart disease, cancer, autoimmunity, genetic defects and even traumatic injury may someday be treated with gene therapy and gene transfer strategies. The potential impact of this new technology on human disease has produced optimism and expectation for scientists and lay people alike. As more effort is directed at harvesting the potential of this technology it has become clear that the success or failure of gene therapy will hinge on our ability to manipulate and control the process of gene transfer in somatic cells. Today, somatic gene transfer is accomplished using either viral or non-viral gene transfer methods. The benefits and limitations of each system are aggressively being investigated to determine which characteristics are most compatible with safe and reliable gene transfer. Gene transfer with cationic lipid/plasmid DNA complexes (cationic lipoplexes) has become a popular means of delivering therapeutic genes and is being tested in preclinical and clinical trials. Cationic lipoplexes are easy and inexpensive to produce, they are composed of non-toxic and non-immunogenic precursor, and they have the potential of delivering large polynucleotides into somatic cells. Additionally, these reagents are easily manipulated in the laboratory to incorporate novel biological functions or to produce new formulations that can be screened for in vivo gene transfer activity. The last few years has seen many advances in our understanding of molecular and biological factors that influence cationic lipid-mediated gene transfer. In this review we discuss recent developments in the field of cationic lipid-mediated gene transfer with emphasis on in vivo application where possible. We will consider new discoveries concerning the molecular and cellular events that control the uptake, transit and expression of lipoplexes in somatic cells. Recent biodistribution and pharmacokinetic studies and current concepts regarding the toxicity and immunogenicity of cationic lipoplexes will also be discussed. We also survey some of the many preclinical and clinical trials using cationic lipid-mediated gene transfer, with emphasis on cancer applications.


The effect of recall, reproduction, and restudy on word learning: a pre-registered study.
Certain manipulations, such as testing oneself on newly learned word associations (recall), or the act of repeating a word during training (reproduction), can lead to better learning and retention relative to simply providing more exposure to the word (restudy). Such benefit has been observed for written words. Here, we test how these training manipulations affect learning of words presented aurally, when participants are required to produce these novel phonological forms in a recall task. Participants (36 English-speaking adults) learned 27 pseudowords, which were paired with 27 unfamiliar pictures. They were given cued recall practice for 9 of the words, reproduction practice for another set of 9 words, and the remaining 9 words were restudied. Participants were tested on their recognition (3-alternative forced choice) and recall (saying the pseudoword in response to a picture) of these items immediately after training, and a week after training. Our hypotheses were that reproduction and restudy practice would lead to better learning immediately after training, but that cued recall practice would lead to better retention in the long term. In all three conditions, recognition performance was extremely high immediately after training, and a week following training, indicating that participants had acquired associations between the novel pictures and novel words. In addition, recognition and cued recall performance was better immediately after training relative to a week later, confirming that participants forgot some words over time. However, results in the cued recall task did not support our hypotheses. Immediately after training, participants showed an advantage for cued Recall over the Restudy condition, but not over the Reproduce condition. Furthermore, there was no boost for the cued Recall condition over time relative to the other two conditions. Results from a Bayesian analysis also supported this null finding. Nonetheless, we found a clear effect of word length, with shorter words being better learned than longer words, indicating that our method was sufficiently sensitive to detect an impact of condition on learning. Our primary hypothesis about training conditions conferring specific advantages for production of novel words presented aurally, especially over long intervals, was not supported by this data. Although there may be practical reasons for preferring a particular method for training expressive vocabulary, no difference in effectiveness was detected when presenting words aurally: reproducing, recalling or restudying a word led to the same production accuracy.


fMRI activity patterns in human LOC carry information about object exemplars within category.
Abstract The lateral occipital complex (LOC) is a set of areas in the human occipito-temporal cortex responding to objects as opposed to low-level control stimuli. Conventional functional magnetic resonance imaging (fMRI) analysis methods based on regional averages could not detect signals discriminative of different types of objects in this region. Here, we examined fMRI signals using multivariate pattern recognition (support vector classification) to systematically explore the nature of object-related information available in fine-grained activity patterns in the LOC. Distributed fMRI signals from the LOC allowed for above-chance discrimination not only of the category but also of within-category exemplars of everyday man-made objects, and such exemplar-specific information generalized across changes in stimulus size and viewpoint, particularly in posterior subregions. Object identity could also be predicted from responses of the early visual cortex, even significantly across the changes in size and viewpoint used here. However, a dissociation was observed between these two regions of interest in the degree of discrimination for objects relative to size: In the early visual cortex, two different sizes of the same object were even better discriminated than two different objects (in accordance with measures of pixelwise stimulus similarity), whereas the opposite was true in the LOC. These findings provide the first evidence that direct evoked fMRI activity patterns in the LOC can be different for individual object exemplars (within a single category). We propose that pattern recognition methods as used here may provide an alternative approach to study mechanisms of neuronal representation based on aspects of the fMRI response independent of those assessed in adaptation paradigms.


The 'fractionable autism triad': a review of evidence from behavioural, genetic, cognitive and neural research.
Autism is diagnosed on the basis of a triad of impairments in social interaction, communication, and flexible imaginative functions (with restricted and repetitive behaviors and interests; RRBIs). There has been a strong presumption that these different features of the syndrome are strongly intertwined and proceed from a common cause at the genetic, cognitive and neural levels. In this review we examine evidence for an alternative approach, considering the triad as largely 'fractionable'. We present evidence from our own twin studies, and review relevant literature on autism and autistic-like traits in other groups. We suggest that largely independent genes may operate on social skills/impairments, communication abilities, and RRBIs, requiring a change in molecular-genetic research approaches. At the cognitive level, we suggest that satisfactory accounts exist for each of the triad domains, but no single unitary account can explain both social and nonsocial features of autism. We discuss the implications of the fractionable-triad approach for both diagnosis and future research directions.


Uptake of antiretroviral therapy and male circumcision after community-based HIV testing and strategies for linkage to care versus standard clinic referral: a multisite, open-label, randomised controlled trial in South Africa and Uganda.
Male circumcision decreases HIV acquisition by 60%, and antiretroviral therapy (ART) almost eliminates HIV transmission from HIV-positive people who are virally suppressed; however, coverage of these interventions has lagged behind targets. We aimed to assess whether community-based HIV testing with counsellor support and point-of-care CD4 cell count testing would increase uptake of ART and male circumcision. We did this multisite, open-label, randomised controlled trial in six research-naive communities in rural South Africa and Uganda. Eligible HIV-positive participants (aged ≥16 years) were randomly assigned (1:1:1) in a factorial design to receive lay counsellor clinic linkage facilitation, lay counsellor follow-up home visits, or standard-of-care clinic referral, and then (1:1) either point-of-care CD4 cell count testing or referral for CD4 testing. HIV-negative uncircumcised men (aged 16-49 years) who could receive secure mobile phone text messages were randomly assigned (1:1:1) to receive text message reminders, lay counsellor visits, or standard clinic referral. The study biostatistician generated the randomisation schedule via a computer-generated random number program with varying block sizes (multiples of six or three) stratified by country. Primary outcomes for HIV-positive people were obtaining a CD4 cell count, linkage to an HIV clinic, ART initiation, and viral suppression at 9 months, and for HIV-negative uncircumcised men were visiting a circumcision facility and uptake of male circumcision at 3 months. We assessed social harms as a safety outcome throughout the study. We did the primary analyses by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT02038582. Between June 6, 2013, and March 11, 2015, 15 332 participants were tested. 2339 (15%) participants tested HIV positive, of whom 1325 (57%) were randomly assigned to receive lay counsellor clinic linkage facilitation (n=437), lay counsellor follow-up home visits (n=449), or standard clinic referral (n=439), and then point-of-care CD4 cell testing (n=206, n=220, and n=213, respectively) or referral for CD4 testing (n=231, n=229, and n=226, respectively). 12 993 (85%) participants tested HIV negative, of whom 750 (6%) uncircumcised men were randomly assigned to receive clinic referral (n=230), text message reminders (n=288), or lay counsellor follow-up visits (n=232). 1218 (93%) of 1303 HIV-positive participants were linked to care, but only 488 (37%) participants initiated ART. Overall, 635 (50%) of 1272 HIV-positive individuals achieved viral suppression at 9 months: 219 (52%) of 419 participants in the clinic facilitation group, 202 (47%) of 431 participants in the lay counsellor follow-up group, and 214 (51%) of 422 participants in the clinic referral group, with no significant differences between groups (p=0·668 for clinic facilitation and p=0·273 for lay counsellor follow-up vs clinic referral). 523 (72%) of 734 HIV-negative men visited a circumcision facility, with no difference between groups. 62 (28%) of 224 men were circumcised in the male circumcision clinic referral group compared with 137 (48%) of 284 men in the text message reminder group (relative risk 1·72, 95% CI 1·36-2·17; p<0·0001) and 106 (47%) of 226 men in the lay counsellor follow-up group (1·67, 1·29-2·14; p=0·0001). No cases of study-related social harm were reported, including probing about partnership separation, unintended disclosure, gender-based violence, and stigma. All the community-based strategies achieved high rates of linkage of HIV-positive people to HIV clinics, roughly a third of whom initiated ART, and of those more than 80% were virally suppressed at 9 months. Uptake of male circumcision was almost two-times higher in men who received text message reminders or lay counsellor visits than in those who received standard-of-care clinic referral. Clinic barriers to ART initiation should be addressed in future strategies to increase the proportion of HIV-positive people accessing treatment and achieving viral suppression. National Institute of Allergy and Infectious Diseases, National Institutes of Health.


Antiviral cytotoxic T-cell memory by vaccination with recombinant Listeria monocytogenes.
Listeria monocytogenes is a facultative intracellular bacterium that is able to escape phagocytic vesicles and replicate in the cytoplasm of infected cells. As with viral vectors, this intracytoplasmic life cycle provides a means for introducing foreign proteins into the major histocompatibility complex class I pathway of antigen presentation. Using recombinant L. monocytogenes (rLM) strains expressing the full-length nucleoprotein (NP) or a single cytotoxic T-lymphocyte (CTL) epitope from lymphocytic choriomeningitis virus (LCMV), we analyzed antiviral CTL responses induced by rLM vaccination. After vaccination, rLM was cleared from the host within 7 days while inducing an LCMV-specific ex vivo CD8+ effector CTL response. Virus-specific CTL memory was maintained for 6 months postvaccination, as demonstrated by vigorous secondary CTL responses after in vitro stimulation. A single immunization with rLM that expressed either the full-length NP gene or the CTL epitope alone resulted in LCMV NP-specific CTL precursor frequencies of approximately 1/10(4) CD8+ T cells. A second rLM vaccination resulted in enhanced virus-specific CTL activity and in vitro proliferation. rLM-vaccinated mice were protected against chronic viral infection by an accelerated virus-specific memory CTL response. These mice cleared infectious virus as well as viral antigen, suggesting that sterilizing immunity was achieved. In contrast to mice that received wild-type LM, rLM-vaccinated mice were protected from virally induced immunosuppression and splenic atrophy associated with chronic LCMV infection. The ability to elicit long-term cell-mediated immunity is fundamental in designing vaccines against intracellular pathogens, and these results demonstrate the efficacy of recombinant LM vaccination for inducing protective antiviral CTL memory.


Therapeutic efficacy of dimethyl fumarate in relapsing-remitting multiple sclerosis associates with ROS pathway in monocytes.
Dimethyl fumarate (DMF) is a first-line-treatment for relapsing-remitting multiple sclerosis (RRMS). The redox master regulator Nrf2, essential for redox balance, is a target of DMF, but its precise therapeutic mechanisms of action remain elusive. Here we show impact of DMF on circulating monocytes and T cells in a prospective longitudinal RRMS patient cohort. DMF increases the level of oxidized isoprostanes in peripheral blood. Other observed changes, including methylome and transcriptome profiles, occur in monocytes prior to T cells. Importantly, monocyte counts and monocytic ROS increase following DMF and distinguish patients with beneficial treatment-response from non-responders. A single nucleotide polymorphism in the ROS-generating NOX3 gene is associated with beneficial DMF treatment-response. Our data implicate monocyte-derived oxidative processes in autoimmune diseases and their treatment, and identify NOX3 genetic variant, monocyte counts and redox state as parameters potentially useful to inform clinical decisions on DMF therapy of RRMS.


Neural Signatures of Prediction Errors in a Decision-Making Task Are Modulated by Action Execution Failures.
Decisions must be implemented through actions, and actions are prone to error. As such, when an expected outcome is not obtained, an individual should be sensitive to not only whether the choice itself was suboptimal but also whether the action required to indicate that choice was executed successfully. The intelligent assignment of credit to action execution versus action selection has clear ecological utility for the learner. To explore this, we used a modified version of a classic reinforcement learning task in which feedback indicated whether negative prediction errors were, or were not, associated with execution errors. Using fMRI, we asked if prediction error computations in the human striatum, a key substrate in reinforcement learning and decision making, are modulated when a failure in action execution results in the negative outcome. Participants were more tolerant of non-rewarded outcomes when these resulted from execution errors versus when execution was successful, but reward was withheld. Consistent with this behavior, a model-driven analysis of neural activity revealed an attenuation of the signal associated with negative reward prediction errors in the striatum following execution failures. These results converge with other lines of evidence suggesting that prediction errors in the mesostriatal dopamine system integrate high-level information during the evaluation of instantaneous reward outcomes.


Epidural analgesia in children: planning, organization and development of a new program.
The goal of the present work is to describe the development and results of the pediatric epidural analgesia program at the ''Hospital Nacional de Pediatria Prof. Dr. J. P. Garrahan'' in Argentina. Patients with thoracotomy, abdominal surgery, osteotomy, amputations or severe trauma were included in the program. The program provided training to the entire staff, control and record of pain treatment and its consequences, 24 h a day availability of anesthesia staff and standard polices and procedures. One hundred fifty children under 16 years of age (median age 11 years, median weight 35 kg) were included in the program during the first 18 months. The median of maximum pain reported during activity was 1 (interquartile range 1 to 4 points) using the Visual Analogue Scale (VAS) or Objective Pain Scale (OPS). Eighty seven children (CI 95% 50% to 67%) presented with postoperative nausea and vomiting, urinary retention, itching, motor blockade or sedation. No patient presented with respiratory depression, hypotension, local anaesthetic toxicity, epidural catheter related infection or death during the program evaluation. The postoperative care program enabled a 98-day reduction in treatment in the intensive care unit. The safe use of pediatric epidural analgesia in general wards may require the careful selection of patients, systematic assessment by trained personnel, training of medical and nursing personnel, clear distribution of responsibilities, use of printed indications, systematic record of pain, sedation and complications, information and education of patients and parents, supply of systems for airway resuscitation and management and continuous quality control and revision of the methods.


PBX3 is a putative biomarker of aggressive prostate cancer.
There is a great need to identify new and better prognostic and predictive biomarkers to stratify prostate cancer patients for optimal treatment. The aims of this study were to characterize the expression profile of pre-B cell leukemia homeobox (PBX) transcription factors in prostate cancer with an emphasis on investigating whether PBX3 harbours any prognostic value. The expression profile of PBX3 and PBX1 in prostate tissue was determined by immunohistochemical and immunoblot analysis. Furthermore, the expression of PBX3 transcript variants was analyzed by RT-PCR, NanoString Technologies®, and by analyzing RNA sequence data. The potential of PBX3 to predict prognosis, either at mRNA or protein level, was studied in four independent cohorts. PBX3 was mainly expressed in the nucleus of normal prostate basal cells, while it showed cytosolic expression in prostatic intraepithelial neoplasia and cancer cells. We detected four PBX3 transcript variants in prostate tissue. Competing risk regression analysis revealed that high PBX3 expression was associated with slower progression to castration resistant prostate cancer (sub-hazard ratio (SHR) 0.18, 95% CI: 0.081-0.42, p values < 0.001). PBX3 expression had a high predictive accuracy (area under the curve (AUC) = 0.82) when combined with Gleason score and age. Patients undergoing radical prostatectomy, with high levels of PBX3 mRNA, had improved prostate cancer specific survival compared to patients expressing low levels (SHR 0.21, 95% CI: 0.46-0.93, p values < 0.001, and AUC = 0.75). Our findings strongly indicate that PBX3 has potential as a biomarker, both as part of a larger gene panel and as an immunohistochemical marker, for aggressive prostate cancer.


Markers of acute toxicity of DDT exposure in pancreatic beta-cells determined by a proteomic approach.
Many compounds have the potential to harm pancreatic beta-cells; organochlorine pollutants belong to those compounds. In this work, we aimed to find markers of acute toxicity of p,p'-DDT exposure among proteins expressed in NES2Y human pancreatic beta-cells employing 2-D electrophoresis. We exposed NES2Y cells to a high concentration (150 μM, LC96 after 72 hours) of p,p'-DDT for 24 and 30 hours and determined proteins with changed expression using 2-D electrophoresis. We have found 22 proteins that changed their expression. They included proteins involved in ER stress (GRP78, and endoplasmin), mitochondrial proteins (GRP75, ECHM, IDH3A, NDUS1, and NDUS3), proteins involved in the maintenance of the cell morphology (EFHD2, TCPA, NDRG1, and ezrin), and some other proteins (HNRPF, HNRH1, K2C8, vimentin, PBDC1, EF2, PCNA, biliverdin reductase, G3BP1, FRIL, and HSP27). The proteins we have identified may serve as indicators of p,p'-DDT toxicity in beta-cells in future studies, including long-term exposure to environmentally relevant concentrations.


Computational procedures for probing interactions in OLS and logistic regression: SPSS and SAS implementations.
Researchers often hypothesize moderated effects, in which the effect of an independent variable on an outcome variable depends on the value of a moderator variable. Such an effect reveals itself statistically as an interaction between the independent and moderator variables in a model of the outcome variable. When an interaction is found, it is important to probe the interaction, for theories and hypotheses often predict not just interaction but a specific pattern of effects of the focal independent variable as a function of the moderator. This article describes the familiar pick-a-point approach and the much less familiar Johnson-Neyman technique for probing interactions in linear models and introduces macros for SPSS and SAS to simplify the computations and facilitate the probing of interactions in ordinary least squares and logistic regression. A script version of the SPSS macro is also available for users who prefer a point-and-click user interface rather than command syntax.


Cross-cultural use and development of virtual patients.
Three major issues drive the cross-cultural use of virtual patients (VPs): an increased mobility of healthcare professionals, students and patients; limited resources for developing VPs; and emerging standards for the exchange of VPs across institutions. Many students are trained in countries other than where they were born. In addition, healthcare professionals often move between countries and are today meeting more and more patients from cultures different from their own. VPs can be used both for learning a new "medical" language as well as for illustrating different perspectives on illness in the new culture. Therefore, it may be important to develop cases reflecting patients from a wide variety of regions and cultures to prepare these professionals to understand both the background of these patients as well as the different medical conditions they may present. However, the benefits of using VPs may be limited at many universities by insufficient resources to develop all the VPs needed for their curricula. The option to acquire VPs from other universities may therefore be appealing, but as these may only be available in English, it is important to consider whether VPs reflecting the local illness panoramas and medical procedures are needed.


A randomized study of a monoclonal antibody (pagibaximab) to prevent staphylococcal sepsis.
Pagibaximab, a human chimeric monoclonal antibody developed against lipoteichoic acid, was effective against staphylococci preclinically and seemed safe and well tolerated in phase 1 studies. To evaluate the clinical activity, pharmacokinetics, safety, and tolerability of weekly pagibaximab versus placebo infusions in very low birth weight neonates. A phase 2, randomized, double-blind, placebo-controlled study was conducted at 10 NICUs. Patients with a birth weight of 700 to 1300 g and 2 to 5 days old were randomly assigned to receive 3 once-a-week pagibaximab (90 or 60 mg/kg) or placebo infusions. Blood was collected for pharmacokinetics, bacterial killing, and safety analyses. Adverse event and clinical outcome data were collected. Eighty-eight patients received pagibaximab at 90 (n = 22) or 60 (n = 20) mg/kg or placebo (n = 46). Groups were not different in demography, mortality, or morbidity. Pagibaximab demonstrated linear pharmacokinetics, a 14.5-day half-life, and nonimmunogenicity. Definite staphylococcal sepsis occurred in 0%, 20%, and 13% (P < .11) and nonstaphylococcal sepsis occurred in 0%, 10%, and 15% (P < .15) of patients in the 90 mg/kg, 60 mg/kg, and placebo groups, respectively. In all patients with staphylococcal sepsis, estimated or observed pagibaximab levels were <500 μg/mL (target level) at infection. Three once-a-week 90 or 60 mg/kg pagibaximab infusions, in high-risk neonates, seemed safe and well tolerated. No staphylococcal sepsis occurred in infants who received 90 mg/kg. Target levels were only consistently achieved after 2 to 3 doses. Dose optimization should enhance protection.


Hypoxia in the thymus: role of oxygen tension in thymocyte survival.
Our previous studies using oxygen microelectrodes showed that the thymus is grossly hypoxic under normal physiological conditions. We now have investigated how oxygen tension affects the thymus at the cellular and molecular level. Adducts of the hypoxia marker drug pimonidazole accumulated in foci within the cortex and medulla and at the corticomedullary junction, consistent with the presence of widespread cellular hypoxia in the normal thymus. Hypoxia-associated pimonidazole accumulation was decreased but not abrogated by oxygen administration. Genes previously reported to be induced by hypoxia were expressed at baseline levels in the normal thymus, indicating that physiological adaptation to hypoxia occurred. Despite changes in thymus size and cellularity, thymic PO(2) did not change with age. Combined assays for hypoxia and cell death showed that hypoxia achieved using either hypoxic gas mixtures or high-density culture in normoxia decreased spontaneous thymocyte apoptosis in vitro. Taken together, these data suggest that regulatory mechanisms exist to maintain thymic cellular hypoxia in vivo and that oxygen tension may regulate thymocyte survival both in vitro and in vivo.


The death of the cancer cell.
For a century, the perception that there are qualitative differences between a normal cell and a cell belonging to a tumor has dominated discussions aimed at explaining cancer. However, an analysis of the experimental evidence suggests that individual normal cells and individual cancer cells share the same two fundamental behavioral properties, namely, proliferation and motility. Each individual cancer cell carries no recognizable molecules or structures that make them consistently distinguishable from normal cells. Herein, we argue that the differences between normal and cancerous states are instead identifiable at the tissue level of biological organization, and therefore, the search for identification of a cancer cell should be abandoned.


A model to measure fluid outflow in rabbit capsules post glaucoma implant surgery.
Prior models of glaucoma filtration surgery assess bleb morphology, which does not always reflect function. Our aim is to establish a model that directly measures tissue hydraulic conductivity of postsurgical outflow in rabbit bleb capsules following experimental glaucoma filtration surgery. Nine rabbits underwent insertion of a single-plate pediatric Molteno implant into the anterior chamber of their left eye. Right eyes were used as controls. The rabbits were then allocated to one of two groups. Group one had outflow measurements performed at 1 week after surgery (n = 5), and group two had measurements performed at 4 weeks (n = 4). Measurements were performed by cannulating the drainage tube ostium in situ with a needle attached to a pressure transducer and a fluid column at 15 mm Hg. The drop in the fluid column was measured every minute for 5 minutes. For the control eyes (n = 6), the anterior chamber of the unoperated fellow eye was cannulated. Animals were euthanized with the implant and its surrounding capsule dissected and fixed in 4% paraformaldehyde, and embedded in paraffin before 6-μm sections were cut for histologic staining. By 7 days after surgery, tube outflow was 0.117 ± 0.036 μL/min/mm Hg at 15 mm Hg (mean ± SEM), whereas at 28 days, it was 0.009 ± 0.003 μL/min/mm Hg. Control eyes had an outflow of 0.136 ± 0.007 μL/min/mm Hg (P = 0.004, one-way ANOVA). Hematoxylin and eosin staining demonstrated a thinner and looser arrangement of collagenous tissue in the capsules at 1 week compared with that at 4 weeks, which had thicker and more densely arranged collagen. We describe a new model to directly measure hydraulic conductivity in a rabbit glaucoma surgery implant model. The principal physiologic endpoint of glaucoma surgery can be reliably quantified and consistently measured with this model. At 28 days post glaucoma filtration surgery, a rabbit bleb capsule has significantly reduced tissue hydraulic conductivity, in line with loss of implant outflow facility, and increased thickness and density of fibrous encapsulation.


The relationship between job stress and mental health at work.
In order to evaluate the relationship between job stress and mental health, a cross-sectional study was conducted using a questionnaire relating to demographics, subjective job stress and mental health state. The questionnaire consisted of a 30-item Japanese version of the General Health Questionnaire (GHQ) developed by Goldberg in addition to questions about subjective job stress, to measure mental health and job stress conditions, respectively. All subjects were employees of an electronic company in Japan. Among 782 workers, 763 workers responded to the questionnaire satisfactorily (response rate was 97.6%). People whose GHQ score was more than 7 were classified as having psychiatric problems, while the remaining respondents were considered as having no mental health problems. We employed a multiple logistic regression analysis to estimate the relationship between subjective job stress and mental health, adjusting for gender, age, marital state, familial stress, and physical health state. Subjective job stress was significantly associated with the state of mental health. In particular, the items of "too much trouble at work," "too much responsibility," "are not allowed to make mistakes," "poor relationship with superiors," and "cannot keep up with technology" were significantly related to mental health.


What impact has tendering had on trainees? Results of a national survey by British Association for Sexual Health and HIV Trainees' Collaborative for audit, research and quality improvement projects.
In April 2013, local authorities gained responsibility for commissioning sexual health services in England. With many services going out to tender and resultant change in services or service provider, there is anecdotal evidence that this has impacted on the education, training and morale of genitourinary medicine (GUM) trainees. The aim of this study was to evaluate the impact of tendering on GUM trainees. An electronic survey designed by the British Association for Sexual Health and HIV Trainees' Collaborative for Audit, Research and Quality Improvement Projects (T-CARQ) was distributed to GUM trainees and newly appointed consultants. Eighty-two individuals responded (74% GUM trainees, 25% newly appointed consultants, 1% locum appointed for service). Sixty-three per cent (45/72) had experience of training within a service which was being tendered. Of these, 59% (24/41) felt their training was not considered during the tendering process and 20% (8/41) felt that it was. Forty-four per cent (18/41) felt adequately supported. Thirty per cent (12/40) reported active participation in the tendering process. On a scale of 0 (no impact) to 5 (major impact), the median score for impact of tendering on training was 2. The positive/negative impact of tendering on different training elements was rated: other than management experience the overall impact on all parameters was negative, namely morale, senior support and education. In conclusion, this survey describes the variable impact of service tendering on GUM training. Our recommendations for maintaining training standards despite tendering include actively involving trainees and education partners, inclusion of specialist GUM training in service specifications, development of guidance for commissioners and services for the management of GUM training within tendering.