Establishment of an index with increased sensitivity for assessing murine arthritis.
The goals of our study were to establish quantitative outcomes for assessing murine knee arthritis and develop an Arthritis Index that incorporates multiple outcomes into a single calculation that provides enhanced sensitivity. Using an accepted model of meniscal/ligamentous injury (MLI)-induced osteoarthritis (OA), we assessed mouse knee arthritis using several approaches. Histology-based methods were performed to visualize joint tissues including articular cartilage and subchondral bone. Accepted histologic scoring methods and histomorphometry were performed to grade cartilage degeneration and determine articular cartilage area, respectively. MicroCT was used to visualize and quantify the bony structures of the joint including osteophytes and joint bone volume. A statistical algorithm was then developed that combined histologic scores and cartilage areas into a single Arthritis Index. MLI induced progressive, OA-like articular cartilage degeneration characterized by increasing (worsening) histologic score and decreasing cartilage area. MicroCT revealed osteophytes and increased joint bone volume between the femoral and tibial physes following MLI. Lastly, an Arthritis Index calculation was established, which incorporated histologic scoring and cartilage area. The Arthritis Index provided enhanced quantitative sensitivity in assessing the level of joint degeneration compared to either histologic scoring or cartilage area determination alone; when using the Index, between 29% and 43% fewer samples are needed to establish statistical significance in studies of murine arthritis. Arthritis in the mouse knee can be quantitatively assessed by histologic scoring, measuring cartilage area, and determining joint bone volume. Enhanced sensitivity can be achieved by performing the Arthritis Index calculation, a novel method for quantitatively assessing mouse knee arthritis.


Fostering trust in healthcare: Participants' experiences, views, and concerns about the 100,000 genomes project.
In this paper, we present findings from a project involving 20 patients with rare diseases, or parents thereof, participating in the 100,000 genomes project (100 kG P). We explored their experiences of, and views about, the project, including why they took part, and their hopes and concerns about the future of genomic medicine. Patients who attended genetic clinics for testing were offered the opportunity to undergo the more extensive whole genome sequencing (WGS) if they agreed to take part in the 100 kG P. Once people had agreed, a specific additional appointment was organised for them. Taking part in the project therefore involved additional travel and appointments ('clinical labour'). We found that interviewees' decisions to participate in 100 kG P were based on interpersonal and institutional trust in the NHS, and on an investment in improving care for the future. Interviewees relied upon receiving good ongoing NHS care for managing their own or their child's rare disease, but they worried about what their relationships with NHS healthcare professionals would be like in future. A few participants worried about whether Genomics England's biorepository would remain protected and an asset of the NHS. To honour and foster participants' trust - which may easily be lost - and their clinical labour, we therefore recommend ongoing public engagement and consultation about how genomics is being integrated more widely across specialties (especially given current funding and staffing constraints in the NHS) within the newly formed NHS Genomic Medicine Service.


An essential role of the cysteine-rich domain of FZD4 in Norrin/Wnt signaling and familial exudative vitreoretinopathy.
The Wnt pathway plays important yet diverse roles in health and disease. Mutations in the Wnt receptor FZD4 gene have been confirmed to cause familial exudative vitreoretinopathy (FEVR). FEVR is characterized by incomplete vascularization of the peripheral retina, which can lead to vitreous bleeding, tractional retinal detachment, and blindness. We screened for mutations in the FZD4 gene in five families with FEVR and identified five mutations (C45Y, Y58C, W226X, C204R, and W496X), including three novel mutations (C45Y, Y58C, and W226X). In the retina, Norrin serves as a ligand and binds to FZD4 to activate the Wnt signaling pathway in normal angiogenesis and vascularization. The cysteine-rich domain (CRD) of FZD4 has been shown to play a critical role in Norrin-FZD4 binding. We investigated the effect of mutations in the FZD4 CRD in Norrin binding and signaling in vitro and in vivo. Wild-type and mutant FZD4 proteins were assayed for Norrin binding and Norrin-dependent activation of the canonical Wnt pathway by cell-surface and overlay binding assays and luciferase reporter assays. In HEK293 transfection studies, C45Y, Y58C, and C204R mutants did not bind to Norrin and failed to transduce FZD4-mediated Wnt/β-catenin signaling. In vivo studies using Xenopus embryos showed that these FZD4 mutations disrupt Norrin/β-catenin signaling as evidenced by decreased Siamois and Xnr3 expression. This study identified a new class of FZD4 gene mutations in human disease and demonstrates a critical role of the CRD in Norrin binding and activation of the β-catenin pathway.


EmtA, a rRNA methyltransferase conferring high-level evernimicin resistance.
Enterococcus faecium strain 9631355 was isolated from animal sources on the basis of its resistance to the growth promotant avilamycin. The strain also exhibited high-level resistance to evernimicin, a drug undergoing evaluation as a therapeutic agent in humans. Ribosomes from strain 9631355 exhibited a dramatic reduction in evernimicin binding, shown by both cell-free translation assays and direct-binding assays. The resistance determinant was cloned from strain 9631355; sequence alignments suggested it was a methyltransferase and therefore it was designated emtA for evernimicin methyltransferase. Evernimicin resistance was transmissible and emtA was localized to a plasmid-borne insertion element. Purified EmtA methylated 50S subunits from an evernimicin-sensitive strain 30-fold more efficiently than those from a resistant strain. Reverse transcription identified a pause site that was unique to the 23S rRNA extracted from resistant ribosomes. The pause corresponded to methylation of residue G2470 (Escherichia coli numbering). RNA footprinting revealed that G2470 is located within the evernimicin-binding site on the ribosome, thus providing an explanation for the reduced binding of the drug to methylated ribosomes.


Nitrogen Arylation for Macrocyclization of Unprotected Peptides.
We describe an efficient and mild method for the synthesis of macrocyclic peptides via nitrogen arylation from unprotected precursors. Various electrophiles and lysine-based nucleophiles were investigated and showed high-yielding product formation, even for a macrocyclization scan with 14 variants. We found that nitrogen-linked aryl products were more stable to base and oxidation when compared to thiol arylated species, thereby highlighting the utility of this methodology. Finally, N-aryl macrocyclization was performed on a p53 peptide inhibitor of MDM2 and resulted in identification of a nanomolar binder with improved proteolytic stability and cell permeability.


How to Determine the Environmental Exposure of PAHs Originating from Biochar.
Biochars are obtained by pyrolyzing biomass materials and are increasingly used within the agricultural sector. Owing to the production process, biochars can contain polycyclic aromatic hydrocarbons (PAHs) in the high mg/kg range, which makes the determination of the environmental exposure of PAHs originating from biochars relevant. However, PAH sorption to biochar is characterized by very high (10(4)-10(6) L/kg) or extreme distribution coefficients (KD) (>10(6) L/kg), which makes the determination of exposure scientifically and technically challenging. Cyclodextrin extractions, sorptive bioaccessibility extractions, Tenax extractions, contaminant traps, and equilibrium sampling were assessed and selected methods used for the determination of bioavailability parameters for PAHs in two model biochars. Results showed that: (1) the KD values of typically 10(6)-10(9) L/kg made the biochars often act as sinks, rather than sources, of PAHs. (2) Equilibrium sampling yielded freely dissolved concentrations (pg-ng/L range) that were below or near environmental background levels. (3) None of the methods were found to be suitable for the direct measurement of the readily desorbing fractions of PAHs (i.e., bioacessibility) in the two biochars. (4) The contaminant-trap method yielded desorption-resistant PAH fractions of typically 90-100%, implying bioaccessibility in the high μg/kg to low mg/kg range.


Amplicon sequencing for the quantification of spoilage microbiota in complex foods including bacterial spores.
Spoilage of food products is frequently caused by bacterial spores and lactic acid bacteria. Identification of these organisms by classic cultivation methods is limited by their ability to form colonies on nutrient agar plates. In this study, we adapted and optimized 16S rRNA amplicon sequencing for quantification of bacterial spores in a canned food matrix and for monitoring the outgrowth of spoilage microbiota in a ready-to-eat food matrix. The detection limit of bar-coded 16S rRNA amplicon sequencing was determined for the number of bacterial spores in a canned food matrix. Analysis of samples from a canned food matrix spiked with a mixture of equinumerous spores from the thermophiles, Geobacillus stearothermophilus and Geobacillus thermoglucosidans, and the mesophiles, Bacillus sporothermodurans, Bacillus cereus, and Bacillus subtilis, led to the detection of these spores with an average limit of 2 × 10(2) spores ml(-1). The data were normalized by setting the number of sequences resulting from DNA of an inactivated bacterial species, present in the matrix at the same concentration in all samples, to a fixed value for quantitative sample-to-sample comparisons. The 16S rRNA amplicon sequencing method was also employed to monitor population dynamics in a ready-to-eat rice meal, incubated over a period of 12 days at 7 °C. The most predominant outgrowth was observed by the genera Leuconostoc, Bacillus, and Paenibacillus. Analysis of meals pre-treated with weak acids showed inhibition of outgrowth of these three genera. The specificity of the amplicon synthesis was improved by the design of oligonucleotides that minimize the amplification of 16S rRNA genes from chloroplasts originating from plant-based material present in the food. This study shows that the composition of complex spoilage populations, including bacterial spores, can be monitored in complex food matrices by bar-coded amplicon sequencing in a quantitative manner. In order to allow sample-to-sample comparisons, normalizations based on background DNA are described. This method offers a solution for the identification and quantification of spoilage microbiota, which cannot be cultivated under standard laboratory conditions. The study indicates variable detection limits among species of bacterial spores resulting from differences in DNA extraction efficiencies.


Plasma protein synthesis by isolated rat hepatocytes.
A system of preparation of rat hepatocytes with extended viability has been developed to study the role of hormones and other plasma components upon secretory protein synthesis. Hepatocytes maintained in minimal essential medium reduced the levels of all amino acids in the medium except the slowly catabolized amino acids leucine, isoleucine, and valine, which steadily increase as the result of catabolism of liver protein. Although the liver cells catabolize 10-15% of their own protein during a 20-h incubation, the cells continue to secrete protein in a linear fashion throughout the period. The effects of insulin, cortisol, and epinephrine on general protein synthesis, and specifically on fibrinogen and albumin synthesis, have been tested on cells from both normal rats and adrenalectomized rats. Cells from normal animals show preinduction of tyrosine amino transferase (TAT), having at the time of isolation a high level of enzyme which shows only an increase of approximately 60% upon incubation with cortisol. In contrast, cells from adrenalectomized animals initially have a low level of enzyme which increases fourfold over a period of 9 h. The effects of both epinephrine and cortisol on protein synthesis are also much larger in cells from adrenalectomized animals. After a delay of several hours, cortisol increases fibrinogen synthesis sharply, so that at the end of the 20-h incubation, cells treated with hormone have secreted nearly 2.5 times as much fibrinogen as control cells. The effect is specific; cortisol stimulates neither albumin secretion nor intracellular protein synthesis. The combination of cortisol and epinephrine strongly depresses albumin synthesis in both types of cells. Insulin enhances albumin and general protein synthesis but has little effect on fibrinogen synthesis.


Little appetite for obesity: meta-analysis of the effects of maternal obesogenic diets on offspring food intake and body mass in rodents.
There is increasing recognition that maternal effects contribute to variation in individual food intake and metabolism. For example, many experimental studies on model animals have reported the effect of a maternal obesogenic diet during pregnancy on the appetite of offspring. However, the consistency of effects and the causes of variation among studies remain poorly understood. After a systematic search for relevant publications, we selected 53 studies on rats and mice for a meta-analysis. We extracted and analysed data on the differences in food intake and body weight between offspring of dams fed obesogenic diets and dams fed standard diets during gestation. We used meta-regression to study predictors of the strength and direction of the effect sizes. We found that experimental offspring tended to eat more than control offspring but this difference was small and not statistically significant (0.198, 95% highest posterior density (HPD)=-0.118-0.627). However, offspring from dams on obesogenic diets were significantly heavier than offspring of control dams (0.591, 95% HPD=0.052-1.056). Meta-regression analysis revealed no significant influences of tested predictor variables (for example, use of choice vs no-choice maternal diet, offspring sex) on differences in offspring appetite. Dietary manipulations that extended into lactation had the largest effect on body weight. Subgroup analysis revealed that high protein to non-protein ratio of the maternal diet may promote increased body weight in experimental offspring in comparison with control offspring; low protein content in the maternal chow can have opposite effect. Exposure to maternal obesogenic diets in early life is not likely to result in a substantial change in offspring appetite. Nevertheless, we found an effect on offspring body weight, consistent with permanent alterations of offspring metabolism in response to maternal diet. Additionally, it appears that protein content of the obesogenic diet and timing of manipulation modulate the effects on offspring body weight in later life.


Frequency of the CHEK2 1100delC mutation among women with breast cancer: an international study.
A founder allele in the CHEK2 gene (1100delC) has been associated with an elevated risk of breast cancer. This allele is responsible for the majority of CHEK2-associated breast cancers in women from northern European countries; however, within Europe, it seems to be rare in countries that are close to the Mediterranean. The frequency of the 1100delC allele has not been measured in non-White populations. We measured the frequency of the CHEK2 founder allele in 3,882 breast cancer patients and 8,609 controls from various countries. The allele was not seen among Asian patients (from Pakistan or the Philippines) and was present in 1 of 155 cases from Brazil. Among White women, the allele was present in 1.5% of 825 familial cases of breast cancer and in 0.7% of 1,106 patients with nonfamilial breast cancer. The allele was equally frequent in Jewish and non-Jewish patients. We estimate that the CHEK2 1100delC allele is associated with an odds ratio of 2.6 for breast cancer, which corresponds to a lifetime risk of approximately 24% in Ontario.


Tactile spatial negative priming occurs without feature mismatch.
Negative priming (NP) refers to fact that people respond more slowly and make more errors when responding to target stimuli that were previously ignored. This phenomenon has also been observed when participants respond to the location, and not only to the identity, of the stimulus. Intriguingly, while roughly the same pattern of results has been observed in the visual, auditory, and tactile modalities when it comes to identity-based NP, the same does not hold true for spatial NP: In particular, feature mismatch seems to be the sole cause of auditory spatial NP, whereas response inhibition would appear to be the sole cause of spatial NP in vision. We conducted a novel tactile variant of the spatial NP task. We investigated whether spatial NP in the tactile modality exists, and further, we investigated whether the pattern of spatial NP in the tactile modality compares with what has been documented previously in vision or audition. Tactile spatial NP was observed, and it was independent of feature mismatch, thereby reflecting a comparable pattern as visual spatial NP. We discuss spatial selection with respect to possible modality-specific processes.


Deactylase inhibitors disrupt cellular complexes containing protein phosphatases and deacetylases.
Affinity isolation of protein serine/threonine phosphatases on the immobilized phosphatase inhibitor microcystin-LR identified histone deacetylase 1(HDAC1), HDAC6, and HDAC10 as novel components of cellular phosphatase complexes. Other HDACs, specifically HDAC2, -3, -4, and -5, were excluded from such complexes. In vitro biochemical studies showed that recombinant HDAC6, but not HDAC4, bound directly to the protein phosphatase (PP)1 catalytic subunit. No association was observed between HDAC6 and PP2A, another major protein phosphatase. PP1 binding was mapped to the second catalytic domain and adjacent C-terminal sequences in HDAC6, and treatment of cells with trichostatin A (TSA) disrupted endogenous HDAC6.PP1 complexes. Consistent with the inhibition of tubulin deactylase activity of HDAC6, TSA enhanced cellular tubulin acetylation, and acetylated tubulin was present in the PP1 complexes from TSA-treated cells. Trapoxin B, a weak HDAC6 inhibitor, and calyculin A, a cell-permeable phosphatase inhibitor, had no effect on the stability of the HDAC6.PP1 complexes or on tubulin acetylation. Mutations that inactivated HDAC6 prevented its incorporation into cellular PP1 complexes and suggested that when bound together both enzymes were active. Interestingly, TSA disrupted all the cellular HDAC.phosphatase complexes analyzed. This study provided new insight into the mechanism by which HDAC inhibitors elicited coordinate changes in cellular protein phosphorylation and acetylation and suggested that changes in these protein modifications at multiple subcellular sites may contribute to the known ability of HDAC inhibitors to suppress cell growth and transformation.


Does Malleolus non-Lifting Tympanoplasty have any Advantage Over Malleus Lifting Techniques?
In order to achieve a higher success rate for tympanoplasty, different techniques have been developed, and a wide variety of grafting materials have been developed. One of the techniques currently receiving considerable attention involves not lifting the remaining of eardrum from the malleus and embedding the graft underneath in order to repair the eardrum correctly in its original position, as well as minimizing graft lateralization leading to progression of hearing rehabilitation. We compared the effects of tympanoplasty with and without malleus lifting on hearing loss in patients with chronic otitis media. In this study, 30 consecutive patients diagnosed as having chronic otitis media without cholesteatoma were randomly assigned to two tympanoplasty groups; with or without malleus lifting. Air and bone conduction thresholds were recorded before and 45 days after the intervention. In groups, except for 8000 Hz, the air conduction was significantly improved following surgery. According to air conduction there was no difference between the groups before surgery at different frequencies, although it was improved to a greater degree in the group without lifting at 250 Hz postoperatively. The average post-operative air-bone gap (ABG) gain was significantly higher in all study frequencies in the target group. One of the effects of this technique is inner-ear protection from physical trauma to the ossicular chain, and prevention of damage to bone conduction. A higher hearing threshold and also higher ABG gain can be achieved by not lifting the remaining eardrum from the malleus and embedding the graft undereath it, especially at lower frequencies.


Trans-cuff portal for arthroscopic posterior capsulorrhaphy.
Arthroscopic repair of posterior shoulder instability is becoming an accepted method of treatment. Most surgeons perform this procedure with the patient in a lateral decubitus position to facilitate access to the posterior glenoid. We have developed an accessory portal placed through the midportion of the rotator cuff that allows easy and complete viewing of the posterior glenohumeral joint when the patient is in a beach chair position and does not require the use of traction. This portal provides a superior-to-inferior view of the posterior glenoid rim and capsule and allows use of anterior and posterior routine portals for posterior Bankart repair. In this study, the technique of posterior labral repair in the beach chair position with use of the trans-cuff portal is described, and preliminary results in 5 patients at an average follow-up of 24 months are presented. Through this approach, we were able to place 3 anchors, with the lowest at the 7 o'clock position (for a right shoulder), in all patients. Mean American Shoulder Elbow Surgeons (ASES) score improved from 53 +/- 15 preoperatively to 87 +/- 8 postoperatively (P < .01). All patients had an excellent result with complete resolution of pain and instability.


MFGE8 does not influence chorio-retinal homeostasis or choroidal neovascularization in vivo.
Milk fat globule-epidermal growth factor-factor VIII (MFGE8) is necessary for diurnal outer segment phagocytosis and promotes VEGF-dependent neovascularization. The prevalence of two single nucleotide polymorphisms (SNP) in MFGE8 was studied in two exsudative or "wet" Age-related Macular Degeneration (AMD) groups and two corresponding control groups. We studied the effect of MFGE8 deficiency on retinal homeostasis with age and on choroidal neovascularization (CNV) in mice. The distribution of the SNP (rs4945 and rs1878326) of MFGE8 was analyzed in two groups of patients with "wet" AMD and their age-matched controls from Germany and France. MFGE8-expressing cells were identified in Mfge8(+/-) mice expressing ß-galactosidase. Aged Mfge8(+/-) and Mfge8(-/-) mice were studied by funduscopy, histology, electron microscopy, scanning electron microscopy of vascular corrosion casts of the choroid, and after laser-induced CNV. rs1878326 was associated with AMD in the French and German group. The Mfge8 promoter is highly active in photoreceptors but not in retinal pigment epithelium cells. Mfge8(-/-) mice did not differ from controls in terms of fundus appearance, photoreceptor cell layers, choroidal architecture or laser-induced CNV. In contrast, the Bruch's membrane (BM) was slightly but significantly thicker in Mfge8(-/-) mice as compared to controls. Despite a reproducible minor increase of rs1878326 in AMD patients and a very modest increase in BM in Mfge8(-/-) mice, our data suggests that MFGE8 dysfunction does not play a critical role in the pathogenesis of AMD.


Current diagnostic problems in melanoma pathology.
The histopathological diagnosis of cutaneous melanocytic lesions may be difficult to assess. Frequently encountered diagnostic problems include: 1) Dysplastic nevus or melanoma in situ?; 2) Melanoma in situ or superficial spreading melanoma?; 3) Lentigo maligna or lentigo maligna melanoma?; 4) Compound nevocellular nevus or nevoid melanoma?; and 5) Spitz nevus or Spitzoid melanoma? Moreover, less frequently encountered diagnostic challenges are discussed: 1) Deep penetrating nevus or nodular melanoma?; and 2) Cellular blue nevus or melanoma metastasis? In this contribution, these problems are discussed after a systematic approach involving a concise histopathological description of the classic lesions considered in the differential diagnoses, a presentation of the deviating histopathological features that give rise to the diagnostic problems, and finally diagnostic recommendations on the classification of the problematic lesions. We also briefly discuss the contribution of additional immunohistochemistry and molecular pathology in aiding to establish a correct diagnosis.


Junk DNA and the long non-coding RNA twist in cancer genetics.
The central dogma of molecular biology states that the flow of genetic information moves from DNA to RNA to protein. However, in the last decade this dogma has been challenged by new findings on non-coding RNAs (ncRNAs) such as microRNAs (miRNAs). More recently, long non-coding RNAs (lncRNAs) have attracted much attention due to their large number and biological significance. Many lncRNAs have been identified as mapping to regulatory elements including gene promoters and enhancers, ultraconserved regions and intergenic regions of protein-coding genes. Yet, the biological function and molecular mechanisms of lncRNA in human diseases in general and cancer in particular remain largely unknown. Data from the literature suggest that lncRNA, often via interaction with proteins, functions in specific genomic loci or use their own transcription loci for regulatory activity. In this review, we summarize recent findings supporting the importance of DNA loci in lncRNA function and the underlying molecular mechanisms via cis or trans regulation, and discuss their implications in cancer. In addition, we use the 8q24 genomic locus, a region containing interactive SNPs, DNA regulatory elements and lncRNAs, as an example to illustrate how single-nucleotide polymorphism (SNP) located within lncRNAs may be functionally associated with the individual's susceptibility to cancer.


First Report of Nerine yellow stripe virus in Amaryllis in the United States.
Ornamental flower bulbs (including true bulbs, bulbils, corms, tubers, and rhizomes) are increasingly important floriculture crops. Amaryllis is a small genus of flowering bulbs, with two species. The South African native, Amaryllis belladonna, also known as belladonna lily, Jersey lily, naked lady, Amarillo, or March lily, is one of numerous ornamental species with the common name "lily" due to their flower shape and growth habit. Amaryllis are popular for their 6- to 10-inch trumpet shaped colorful flowers that are borne on 1- to 2-foot stalks. In January, 2011, a home gardener in California observed mosaic symptoms on the leaves of A. belladonna growing in her garden. Leaf samples were sent to Agdia Inc. for testing. Samples tested positive for the presence of Potyvirus in a reverse transcription (RT)-PCR screen using universal potyvirus primers (2) yielding the expected ∼1,600-bp product corresponding to the partial nuclear inclusion body (NIb) gene, full-length coat protein (CP) gene, and 3' end untranslated region (UTR). Electron microscopy of symptomatic leaves confirmed the presence of filamentous potyvirus-like particles. The RT-PCR amplicon was cloned and sequenced (2); the 1,616-bp consensus sequence was deposited in GenBank (Accession No. JX865782). NCBI BLAST analysis of the consensus sequence revealed highest identities with isolates of Nerine yellow stripe virus (NeYSV; family Potyviridae, genus Potyvirus). Pair-wise analyses of the 261 amino acid sequence of the predicted CP had 88% sequence identity with a Stenomesson isolate reported from the Netherlands (EU042758); 87% identity with Hymenocallis and Nerine isolates, both also from the Netherlands (EF362622 and EF362621, respectively); and, 86% with two New Zealand isolates infecting Amaryllis or Vallota (FJ618537 and DQ407932, respectively). The five Netherlands and New Zealand isolates are more closely related to each other than to the U.S. isolate as they share 93 to 98% CP identity. When using viral genome sequence relatedness as a criterion for defining potyvirus species, isolates with CP amino acid identity greater than 80% are considered the same species (1). The predicted coat protein gene of the California isolate was sub-cloned into the bacterial expression vector pET44 EK/LIC. Serological analysis of coat protein expressing clones in ELISA and Western Blot analysis using a potyvirus broad-spectrum reacting monoclonal antibody PTY-2 (3) and a NeYSV-specific rabbit antiserum (Applied Plant Research, Lisse, The Netherlands) resulted in positive reactions. NeYSV has previously been reported in the United Kingdom, the Netherlands, Australia, and New Zealand. Based on the results of electron microscopy, RT-PCR, nucleotide and amino acid identity, and serological reactivity, we identify this virus as a U.S. isolate of NeYSV, NeYSV-US. To our knowledge, this is the first report of Nerine yellow stripe virus in the United States. Development of antisera specific to this U.S. isolate is in progress. References: (1) A. Gibbs and K. Ohshima. Ann. Rev. Phytopathol. 48:205, 2010. (2) R. L. Jordan et al. Acta Hortic. 901:159, 2011. (3) R. L. Jordan and J. Hammond. J. Gen. Virol. 72:1531, 1991.


Low-dose ketamine adjuvant treatment for refractory pain in children, adolescents and young adults with cancer: a pilot study.
Ketamine, an N-methyl-D-aspartate receptor antagonist, is effective at relieving adult cancer pain, although there have been very few reports to date regarding its use in children and in adolescents and young adults (AYA). This study assessed the efficacy, safety and opioid-sparing effects of low doses of ketamine added to opioid analgesics to alleviate persistent cancer pain. This prospective, multicentre, observational trial collected data regarding demographics, pain characteristics, pain score assessment within the first 48 hours of ketamine administration, tolerance and satisfaction from 38 patients aged 2-24 years prescribed with ketamine as an adjuvant antalgic for refractory cancer pain in 10 French paediatric oncology centres. The mean visual analogue scale pain score decreased from 6.7 to 4.3 out of 10 (n=39, p<0.001) from day 1 to day 3 and by at least 2 points in 56% of the patients (n=22) 48 hours after initiation of ketamine. Nine patients experienced poor tolerance (≥2 side effects), all with infusion rates lower than 0.05 mg/kg/hour. None had limiting toxicities. An opioid-sparing effect was highlighted in four patients. Fifty-four per cent of the prescribers and 47% of the patients found the addition of ketamine 'very helpful'. Low doses of ketamine as an adjuvant to opioids significantly reduced the intensity of pain in half of the study population. A tendency towards better pain control is shown, although a lack of statistical power somewhat limits our conclusions, especially in children. Nevertheless, ketamine may be a useful option for improving the treatment of refractory pain in children and AYA with cancer.


Senescence and apoptosis block hematopoietic activation of quiescent hematopoietic stem cells with short telomeres.
Telomere shortening limits the proliferative capacity of human cells, and age-dependent shortening of telomeres occurs in somatic tissues including hematopoietic stem cells (HSCs). It is currently unknown whether genomic and molecular damage that occurs in HSCs induced by telomere shortening is transmitted to the progenitor cells. Here we show that telomere shortening results in DNA damage accumulation and gene expression changes in quiescent HSCs of aged mice. Upon activation, a subset of HSCs with elevated levels of DNA damage and p16 expression are blocked from cell cycle entry, and apoptosis is induced in HSCs entering the cell cycle. Activation of both checkpoints associates with normalization of DNA damage and gene expression profiles at early progenitor stages. These findings indicate that quiescent HSCs have an elevated tolerance to accumulate genomic alterations in response to telomere shortening, but the transmission of these aberrations to the progenitor cell level is prevented by senescence and apoptosis.


Protection against amyloid beta cytotoxicity by sulforaphane: role of the proteasome.
The 26S proteasome plays a major role in degradation of abnormal proteins within the cell. The indirect antioxidant including sulforaphane (SFN) protects cells from oxidative damage by increasing the expression of Nrf2-target genes. It has been observed that the expression of multiple subunits of the proteasome was up-regulated by indirect antioxidants through the Nrf2 pathway. In the current study, the role of SFN in amyloid beta(1-42) (Abeta(1-42))-induced cytotoxicity has been investigated in murine neuroblastoma cells. Treatment with SFN protected cells from Abeta(1-42)-mediated cell death in Neuro2A and N1E 115 cells. Inhibition of proteasome activities by MG132 could abolish the protective effect of SFN against Abeta(1-42). Neuro2A cells, which were stably overexpressing the catalytic subunit of the proteasome PSMB5, showed an elevated resistance toward Abeta(1-42) toxicity compared to control cells. Furthermore, the in vitro assay demonstrated that the Abeta(1-42) peptide is degraded by the proteasome fraction. These results suggest that proteasome-inducing indirect antioxidants may facilitate the removal of the Abeta(1-42) peptide and lead to the amelioration of abnormal protein-associated etiologies.


Adaptive female-mimicking behavior in a scorpionfly.
This study provides a clear example of female-mimicking behavior by males in insects and evaluates quantitatively the adaptive significance of this behavior, which is poorly understood in many other organisms. Males of Hylobittacus apicalis provide females with a prey arthropod during copulation. Some males mimic female behavior when interacting with males that have captured nuptial prey, resulting in males stealing prey which they will use for copulation. Males that pirate prey copulate more frequently and probably incur fewer predation-related risks.


Optically controlled loss in an optical fiber.
The use of upconversion to attenuate/modulate optical signals is proposed, and experimental results are presented. In this light controlling light approach, a 711.2 nm, 700 mW beam propagating in a Tm(3+)-doped fluorozirconate optical fiber was attenuated down to 150 mW by copropagating a 1117 nm beam. Further characterizations concerning time response of the transmittance revealed the limits and potential of the scheme. The maximum attenuation of a 45 cm fiber, 75%, was reached up to 400 Hz, and at least 37% modulation was measured at 770 Hz. Potential applications include Q switching of lasers and in-fiber light-controllable optical attenuation.


Multiple electron capture from isolated protein poly-anions in collision with slow highly charged ions.
Collisions of 375 keV Xe25+ ions with trapped mass/charge selected poly-anions of the cytochrome C protein (∼12.5 kDa) were studied by coupling a linear quadrupole ion trap with low-energy ion beam facility. Tandem mass spectra were recorded for the protein precursor charge states ranging from -9 to -17. The present work reports the first study of slow highly charged ion collisions with poly-anions. A high signal to noise ratio allowed the study of the intensity of single and multiple electron removal by a projectile, as well as associated neutral losses, as a function of the target charge state. Relative single and double electron detachment cross sections were found to increase with increasing charge state of the precursor anion. The experimental findings are supported by the calculations of the total electron capture cross sections, based on the classical over-the-barrier model, restricted to a simple uniformly charged linear protein structure and a near-end electron capture.


A pilot study on the refinement of acute inhalation toxicity studies: the isolated perfused rat lung as a screening tool for surface-active substances.
New surface-active agents in waterproofing sprays are frequently tested for acute inhalation toxicity in vivo according to OECD Test Guideline 403. In order to refine and reduce the number of acute inhalation tests performed, we propose a screening test that uses isolated lungs. The test consists of the exposure of isolated, ventilated and perfused rat lungs, to aerosolised formulations of waterproofing agents (mass median aerodynamic diameter = 1μm), and on-line monitoring of respiratory parameters and gross pathology analysis. A pilot evaluation of the isolated perfused rat lung model for use in a screening test was carried out by blind testing 12 surface-active substances. The results obtained compared well with data available from in vivo acute inhalation studies. Substances that triggered harmful effects, such as impaired lung compliance and atelectasis of the isolated perfused lung, were also found to cause changes in respiratory parameters, some of which would be severe enough to lead to death in in vivo tests with rats. The changes in respiratory parameters suggest that the mode-of-action is associated with impairment of the surfactant layer. Therefore, pre-testing in the isolated perfused rat lung allows the identification of surface-active substances with the potential for causing acute inhalation toxicity.


USP9X Deubiquitylates DVL2 to Regulate WNT Pathway Specification.
The WNT signaling network is comprised of multiple receptors that relay various input signals via distinct transduction pathways to execute multiple complex and context-specific output processes. Integrity of the WNT signaling network relies on proper specification between canonical and noncanonical pathways, which presents a regulatory challenge given that several signal transducing elements are shared between pathways. Here, we report that USP9X, a deubiquitylase, and WWP1, an E3 ubiquitin ligase, regulate a ubiquitin rheostat on DVL2, a WNT signaling protein. Our findings indicate that USP9X-mediated deubiquitylation of DVL2 is required for canonical WNT activation, while increased DVL2 ubiquitylation is associated with localization to actin-rich projections and activation of the planar cell polarity (PCP) pathway. We propose that a WWP1-USP9X axis regulates a ubiquitin rheostat on DVL2 that specifies its participation in either canonical WNT or WNT-PCP pathways. These findings have important implications for therapeutic targeting of USP9X in human cancer.


Reproducibility, stability, and accuracy of microbial profiles by fecal sample collection method in three distinct populations.
The gut microbiome likely plays a role in the etiology of multiple health conditions, especially those affecting the gastrointestinal tract. Little consensus exists as to the best, standard methods to collect fecal samples for future microbiome analysis. We evaluated three distinct populations (N = 132 participants) using 16S rRNA gene amplicon sequencing data to investigate the reproducibility, stability, and accuracy of microbial profiles in fecal samples collected and stored via fecal occult blood test (FOBT) or Flinders Technology Associates (FTA) cards, fecal immunochemical tests (FIT) tubes, 70% and 95% ethanol, RNAlater, or with no solution. For each collection method, based on relative abundance of select phyla and genera, two alpha diversity metrics, and four beta diversity metrics, we calculated intraclass correlation coefficients (ICCs) to estimate reproducibility and stability, and Spearman correlation coefficients (SCCs) to estimate accuracy of the fecal microbial profile. Comparing duplicate samples, reproducibility ICCs for all collection methods were excellent (ICCs ≥75%). After 4-7 days at ambient temperature, ICCs for microbial profile stability were excellent (≥75%) for most collection methods, except those collected via no-solution and 70% ethanol. SCCs comparing each collection method to immediately-frozen no-solution samples ranged from fair to excellent for most methods; however, accuracy of genus-level relative abundances differed by collection method. Our findings, taken together with previous studies and feasibility considerations, indicated that FOBT/FTA cards, FIT tubes, 95% ethanol, and RNAlater are excellent choices for fecal sample collection methods in future microbiome studies. Furthermore, establishing standard collection methods across studies is highly desirable.


Simultaneous identification of multiple driver pathways in cancer.
Distinguishing the somatic mutations responsible for cancer (driver mutations) from random, passenger mutations is a key challenge in cancer genomics. Driver mutations generally target cellular signaling and regulatory pathways consisting of multiple genes. This heterogeneity complicates the identification of driver mutations by their recurrence across samples, as different combinations of mutations in driver pathways are observed in different samples. We introduce the Multi-Dendrix algorithm for the simultaneous identification of multiple driver pathways de novo in somatic mutation data from a cohort of cancer samples. The algorithm relies on two combinatorial properties of mutations in a driver pathway: high coverage and mutual exclusivity. We derive an integer linear program that finds set of mutations exhibiting these properties. We apply Multi-Dendrix to somatic mutations from glioblastoma, breast cancer, and lung cancer samples. Multi-Dendrix identifies sets of mutations in genes that overlap with known pathways - including Rb, p53, PI(3)K, and cell cycle pathways - and also novel sets of mutually exclusive mutations, including mutations in several transcription factors or other genes involved in transcriptional regulation. These sets are discovered directly from mutation data with no prior knowledge of pathways or gene interactions. We show that Multi-Dendrix outperforms other algorithms for identifying combinations of mutations and is also orders of magnitude faster on genome-scale data. Software available at: http://compbio.cs.brown.edu/software.


Identification of genes in the tomato big bud phytoplasma and comparison to those in sweet potato little leaf-V4 phytoplasma.
Genetic relatedness of phytoplasmas is commonly defined on the basis of differences in the highly conserved 16S rRNA gene, which may not resolve closely related phytoplasmas. An example of this is the closely related tomato big bud (TBB) and sweet potato little leaf strain V4 (SPLL-V4) phytoplasmas, which cannot easily be differentiated by their 16S rRNA gene sequences. This study aimed to identify genes on the TBB phytoplasma chromosome which could be used to examine genetic variation between these two closely related phytoplasmas. Random clones generated from TBB phytoplasma genomic DNA were sequenced and characterized by database analysis. Twenty-three genes were identified within 19 random clones, which contained approximately 18.0 kbp of TBB phytoplasma genomic DNA. Half of the TBB phytoplasma genes identified were involved in DNA replication, transcription and translation. The remaining TBB phytoplasma genes were involved in protein secretion, cellular processes and energy metabolism. Phylogenetic analysis of representative genes showed that the TBB phytoplasma grouped with the mycoplasmas with the exception of the TBB phytoplasma secA gene, which grouped with the onion yellows phytoplasma. PCR primers were designed based on the new genes and tested on isolates of the TBB and SPLL-V4 phytoplasmas. Most primers amplified a product from TBB and SPLL-V4 phytoplasma samples. When amplified products were subjected to RFLP analysis, the restriction patterns were the same as the respective original clones. This result confirmed that the same sequence had been amplified by PCR and showed that these isolates were indistinguishable using the new genes. This study showed that in fact the TBB and SPLL-V4 phytoplasmas are closely related even with the analysis of new genes. These new genes have, however, provided insight into the biology of the TBB and SPLL-V4 phytoplasmas.


Biocompatibility of intraocular lens power adjustment using a femtosecond laser in a rabbit model.
To evaluate the biocompatibility (uveal and capsular) of intraocular lens (IOL) power adjustment by a femtosecond laser obtained through increased hydrophilicity of targeted areas within the optic, creating the ability to build a refractive-index shaping lens within an existing IOL. John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. Experimental study. Six rabbits had phacoemulsification with bilateral implantation of a commercially available hydrophobic acrylic IOL. The postoperative power adjustment was performed 2 weeks after implantation in 1 eye of each rabbit. The animals were followed clinically for an additional 2 weeks and then killed humanely. Their globes were enucleated and bisected coronally just anterior to the equator for gross examination from the Miyake-Apple view to assess capsular bag opacification. After IOL explantation for power measurements, the globes were sectioned and processed for standard histopathology. Slitlamp examinations performed after the laser treatments showed the formation of small gas bubbles behind the lenses that disappeared within a few hours. No postoperative inflammation or toxicity was observed in the treated eyes, and postoperative outcomes and histopathological examination results were similar to those in untreated eyes. The power measurements showed that the change in power obtained was consistent and within ±0.1 diopter of the target. Consistent and precise power changes can be induced in the optic of commercially available IOLs in vivo by using a femtosecond laser to create a refractive-index shaping lens. The laser treatment of the IOLs was biocompatible.


Genetic determinants of aromatase inhibitor-related arthralgia: the B-ABLE cohort study.
A major side effect of aromatase inhibitor (AI) therapy is AI-related arthralgia (AIA), which often leads to therapy discontinuation. We aimed to identify genetic variants associated with AIA and therapy discontinuation in the first year of AI treatment. Our prospective cohort study included 343 postmenopausal women with early breast cancer starting AI therapy. Single nucleotide polymorphisms (SNPs) in candidate genes involved in estrogen and vitamin D signaling were selected. Univariate and multivariate linear/logistic regressions were fitted in order to asses the association between studied SNPs and AIA intensity (visual analogic scale score) at 3 and 12 months of follow-up, worsening pain, and therapy discontinuation. We also tested for a priori-defined interactions by introducing multiplicative terms in the regression equations. SNPs in CYP17A1 and VDR genes appeared significantly associated with AIA (P = 0.003, P = 0.012, respectively). One SNP in CYP27B1 gene was related to therapy discontinuation [P = 0.02; OR 0.29 (0.09-0.99)]. We revealed interactions between CYP27B1 and both CYP17A1 (P = 0.01) and VDR SNPs (P = 0.06). Furthermore, an additive effect on pain intensity was shown for unfavorable alleles, with two points higher mean absolute pain increase and up to 5.3-fold higher risk of worsening pain compared to favorable genotypes. SNPs in CYP17A1, VDR, and CYP27B1 genes predict the risk of AIA. Their determination would be useful to trigger the monitoring strategies in women at risk of therapy discontinuation.


Potential Therapeutic Value of a Novel FAAH Inhibitor for the Treatment of Anxiety.
Anxiety disorders are among the most prevalent psychiatric diseases with high personal costs and a remarkable socio-economic burden. However, current treatment of anxiety is far from satisfactory. Novel pharmacological targets have emerged in the recent years, and attention has focused on the endocannabinoid (eCB) system, given the increasing evidence that supports its central role in emotion, coping with stress and anxiety. In the management of anxiety disorders, drug development strategies have left apart the direct activation of type-1 cannabinoid receptors to indirectly enhance eCB signalling through the inhibition of eCB deactivation, that is, the inhibition of the fatty acid amide hydrolase (FAAH) enzyme. In the present study, we provide evidence for the anxiolytic-like properties of a novel, potent and selective reversible inhibitor of FAAH, ST4070, orally administered to rodents. ST4070 (3 to 30 mg/kg per os) administered to CD1 male mice induced an increase of time spent in the exploration of the open arms of the elevated-plus maze. A partial reduction of anxiety-related behaviour by ST4070 was also obtained in Wistar male rats, which moderately intensified the time spent in the illuminated compartment of the light-dark box. ST4070 clearly inhibited FAAH activity and augmented the levels of two of its substrates, N-arachidonoylethanolamine (anandamide) and N-palmitoylethanolamine, in anxiety-relevant brain regions. Altogether, ST4070 offers a promising anxiolytic-like profile in preclinical studies, although further studies are warranted to clearly demonstrate its efficacy in the clinic management of anxiety disorders.


Dynamic measurements of CO diffusing capacity using discrete samples of alveolar gas.
It has been shown that measurements of the diffusing capacity of the lung for CO made during a slow exhalation [DLCO(exhaled)] yield information about the distribution of the diffusing capacity in the lung that is not available from the commonly measured single-breath diffusing capacity [DLCO(SB)]. Current techniques of measuring DLCO(exhaled) require the use of a rapid-responding (less than 240 ms, 10-90%) CO meter to measure the CO concentration in the exhaled gas continuously during exhalation. DLCO(exhaled) is then calculated using two sample points in the CO signal. Because DLCO(exhaled) calculations are highly affected by small amounts of noise in the CO signal, filtering techniques have been used to reduce noise. However, these techniques reduce the response time of the system and may introduce other errors into the signal. We have developed an alternate technique in which DLCO(exhaled) can be calculated using the concentration of CO in large discrete samples of the exhaled gas, thus eliminating the requirement of a rapid response time in the CO analyzer. We show theoretically that this method is as accurate as other DLCO(exhaled) methods but is less affected by noise. These findings are verified in comparisons of the discrete-sample method of calculating DLCO(exhaled) to point-sample methods in normal subjects, patients with emphysema, and patients with asthma.


Highlights in nonhymenoptera anaphylaxis.
The purpose of this review is to summarize new interesting findings in both research and clinical practice of nonhymenoptera anaphylaxis with particular emphasis on food and drug allergy. Recent findings include some aspects of crossreactivity among betalactams, nonsteroidal anti-inflammatory drugs with special attention to an immunogenic metabolite of diclofenac and the mode of diclofenac-induced basophil degranulation. Also, new insights into the sometimes atypical anaphylaxis-like reactions after administration of monoclonal antibodies and the immunologic response to these molecules will be discussed, as well as the role of low-dose pholcodine exposure on the development of sensitization to neuromuscular blocking agents. In food-related anaphylaxis, a significant increase of incidents has been observed over the last few years, especially among children of less than 5 years of age. Unfortunately, the severity of subsequent reactions cannot be predicted from the reaction history. Tree nuts and peanuts are the most important elicitors of fatal food-induced anaphylaxis. In food-related anaphylaxis, prospectively evaluated marker allergens that are indicative of a potential anaphylactic reaction are still lacking. The drug groups covered in this review are of special interest either due to their prominent role as allergens or due to new findings.


Optimal FDG PET/CT volumetric parameters for risk stratification in patients with locally advanced non-small cell lung cancer: results from the ACRIN 6668/RTOG 0235 trial.
In recent years, multiple studies have demonstrated the value of volumetric FDG-PET/CT parameters as independent prognostic factors in patients with non-small cell lung cancer (NSCLC). We aimed to determine the optimal cut-off points of pretreatment volumetric FDG-PET/CT parameters in predicting overall survival (OS) in patients with locally advanced NSCLC and to recommend imaging biomarkers appropriate for routine clinical applications. Patients with inoperable stage IIB/III NSCLC enrolled in ACRIN 6668/RTOG 0235 were included. Pretreatment FDG-PET scans were quantified using semiautomatic adaptive contrast-oriented thresholding and local-background partial-volume-effect-correction algorithms. For each patient, the following indices were measured: metabolic tumor volume (MTV), total lesion glycolysis (TLG), SUVmax, SUVmean, partial-volume-corrected TLG (pvcTLG), and pvcSUVmean for the whole-body, primary tumor, and regional lymph nodes. The association between each index and patient outcome was assessed using Cox proportional hazards regression. Optimal cut-off points were estimated using recursive binary partitioning in a conditional inference framework and used in Kaplan-Meier curves with log-rank testing. The discriminatory ability of each index was examined using time-dependent receiver operating characteristic (ROC) curves and corresponding area under the curve (AUC(t)). The study included 196 patients. Pretreatment whole-body and primary tumor MTV, TLG, and pvcTLG were independently prognostic of OS. Optimal cut-off points were 175.0, 270.9, and 35.5 cm3 for whole-body TLG, pvcTLG, and MTV, and were 168.2, 239.8, and 17.4 cm3 for primary tumor TLG, pvcTLG, and MTV, respectively. In time-dependent ROC analysis, AUC(t) for MTV and TLG were uniformly higher than that of SUV measures over all time points. Primary tumor and whole-body parameters demonstrated similar patterns of separation for those patients above versus below the optimal cut-off points in Kaplan-Meier curves and in time-dependent ROC analysis. We demonstrated that pretreatment whole-body and primary tumor volumetric FDG-PET/CT parameters, including MTV, TLG, and pvcTLG, are strongly prognostic for OS in patients with locally advanced NSCLC, and have similar discriminatory ability. Therefore, we believe that, after validation in future trials, the derived optimal cut-off points for primary tumor volumetric FDG-PET/CT parameters, or their more refined versions, could be incorporated into routine clinical practice, and may provide more accurate prognostication and staging based on tumor metabolic features.


Chronic depression and comorbid personality disorders: response to sertraline versus imipramine.
Chronic subtypes of depression appear to be associated with high rates of Axis II personality disorder comorbidity. Few studies, though, have systematically examined the clinical correlates of Axis II personality disorder comorbidity or its effect on treatment response or time to response. 635 patients diagnosed with DSM-III-R chronic major depression or "double depression" (dysthymia with concurrent major depression) were randomized to 12 weeks of double-blind treatment with either sertraline or imipramine between February 1993 and December 1994. Axis II diagnoses were made using the personality disorders version of the DSM-III-R Structured Clinical Interview. The effect of study treatment was measured utilizing the Hamilton Rating Scale for Depression and the Clinical Global Impressions scale. Forty-six percent of patients met criteria for at least 1 comorbid Axis II personality disorder, with cluster C diagnoses being the most frequent at 39%; 21% met criteria for at least 2 Axis II personality disorders. A cluster C diagnosis was associated with significantly higher rates of early-onset depression (before age 21; 47% vs. 32% for no cluster C; p =.005) and comorbid anxiety disorder (34% vs. 18% for no cluster C; p <.001). Overall, the presence of Axis II personality disorder comorbidity had minimal-to-no effect on the ability to achieve either an antidepressant response or remission and had inconsistent effects on time to response. The presence of Axis II personality disorder comorbidity did not appear to reduce functional and quality-of-life improvements among patients responding to acute treatment with sertraline or imipramine. In this treatment sample, rates of Axis II personality disorder comorbidity were substantial in patients suffering from chronic forms of depression. Axis II personality disorder comorbidity did not appear to diminish symptomatic response to acute treatment or associated improvement in functioning and quality of life.


The Role of Gender in Neuropsychological Assessment in Healthy Adolescents.
Research in college athletes has revealed significant gender differences in areas of verbal memory, visual memory, and reaction time. Additionally, research has focused on differences in neuropsychological components and gender in college populations; however, such differences in gender have not been documented in healthy adolescent populations. To identify potential differences between males and females using different components of a common computerized neuropsychological test. A computerized neuropsychological test battery (ImPACT®) was administered to 662 high-school age adolescent athletes (male: n = 451 female: n = 262). Differences between genders were calculated using a 1-way ANOVA. All statistical analyses were conducted using SPSS 23.0. Significance levels were set a priori at P < .05. A 1-way ANOVA revealed statistically significant differences between males and females for composite reaction time (F1,660 = 10.68, P = .001) and total symptom score (F1,660 = 81.20, P < .001). However, no statistically significant differences were found between males and females in composite verbal memory, visual memory, visual motor, or impulse control (P > .05). Significant differences between males and females were discovered for composite reaction time and total symptom scores, with females reporting more symptoms and slower reaction times at a baseline assessment. Increased symptom reporting by females may be attributed to both hormonal differences and increased honesty. Quicker reaction times in males may support theories that repetition of activities and quicker muscle contraction are gender dependent. However, additional research is necessary to understand gender differences in adolescent athletes during periods of cognitive and physical maturation.


Loss of endocytic capacity in aging Paramecium. The importance of cytoplasmic organelles.
Aged cells have significantly fewer food vacuoles and ingest fewer bacteria than young cells. Loss of food vacuoles was explained by a decreasing difference in the food vacuole formation and excretion rates; the formation rate declined more rapidly than the excretion rate, approaching equivalence at 160 fissions, when the proportion of cells with no food vacuoles, in the presence of excess food, abruptly increased. A model for cellular aging is presented in which control of organelle numbers and cyclical interactions between the nucleus and cytoplasm may be of critical importance.


Polymerization led selective detection and removal of Zn2+ and Cd2+ ions: isolation of Zn- and Cd-MOFs and reversibility studies.
This work illustrates two post-functionalized chemosensors and their remarkable role in the sensing of Zn2+ and Cd2+ ions via producing the corresponding insoluble metal-organic frameworks (MOFs). The structural characterization of the resultant Zn- and Cd-MOFs revealed their polymeric one- or two-dimensional architectures. Importantly, nitrate and nitrite ions reversed the MOF-polymerization process and regenerated the chemosensors and the corresponding metal salts. These results are significant in terms of recyclability and remediation processes for the removal of metals both from living organisms and the environment.


Effects of changing immunosuppressive regimen on the incidence, duration, and viral load of cytomegalovirus infection in renal transplantation: a single center report.
Background. In this retrospective single center study we have evaluated the relation between the immunosuppressive regimen and the incidence and characteristics of cytomegalovirus (CMV) infection in the setting without CMV prophylaxis from 1989 through 1998. Methods. All (470) first cadaveric renal transplantations in nonsensitized (PRA < 60%) patients were analyzed. Immunosuppression consisted of cyclosporine A (Sandimmune) and prednisolone from 1989 through 2-1993 (S; 189 patients), of cyclosporine microemulsion (Neoral) and prednisolone from 3-1993 through 5-1997 (N; 200 patients) and of mycophenolate mofetil, Neoral and prednisolone from 5-1997 until 1998 (M; 81 patients). The CMV pp65-antigenemia was measured routinely at least once weekly from day 10 till 12 weeks after transplantation or until pp65-antigenemia became negative. No CMV-prophylaxis was given. Results. By changing from Sandimmune to Neoral and by adding mycophenolate mofetil, respectively, we observed a higher frequency of especially secondary CMV infections (S vs. N vs. M, respectively, 28 vs. 50 vs. 63%, P = 0.026; S vs. N, P = 0.027; S vs. M, P = 0.015; and N vs. M, n.s). The CMV infections lasted longer (median duration antigenemia S vs. N vs. M, respectively, 3 vs. 5 vs. 7 weeks, P = 0.0003; S vs. N, P < 0.002; S vs. M, P < 0.001; and N vs. M, P < 0.05). Viral load was higher in M (median maximal pp65-antigenemia S vs. N vs. M, respectively, 19 vs. 14.5 vs. 73, P < 0.01; S vs. N, n.s.; S vs. M, P < 0.001 and N vs. M, P < 0.01). Conclusions. The use of Neoral and the addition of mycophenolate mofetil caused significant changes in the incidence, duration and viral load of CMV infections.


3D plasmonic chiral colloids.
3D plasmonic chiral colloids are synthesized through deterministically grouping of two gold nanorod AuNRs on DNA origami. These nanorod crosses exhibit strong circular dichroism (CD) at optical frequencies which can be engineered through position tuning of the rods on the origami. Our experimental results agree qualitatively well with theoretical predictions.


The role of synaptic GTPase-activating protein in neuronal development and synaptic plasticity.
Synaptic GTPase-activating protein (SynGAP) is a neuronal RasGAP (Ras GTPase-activating protein) that is selectively expressed in brain and highly enriched at excitatory synapses, where it negatively regulates Ras activity and its downstream signaling pathways. To investigate the physiological role of SynGAP in the brain, we have generated mutant mice lacking the SynGAP protein. These mice exhibit postnatal lethality, indicating that SynGAP plays a critical role during neuronal development. In addition, cell biological experiments show that neuronal cultures from mutant mice have more synaptic AMPA receptor clusters, suggesting that SynGAP regulates glutamate receptor synaptic targeting. Moreover, electrophysiological studies demonstrated that heterozygous mutant mice have a specific defect in hippocampal long-term potentiation (LTP). These studies show that the regulation of synaptic Ras signaling by SynGAP is important for proper neuronal development and glutamate receptor trafficking and is critical for the induction of LTP.


Effectiveness of teaching evidence-based medicine to undergraduate medical students: a BEME systematic review.
Despite the widespread teaching of evidence-based medicine (EBM) to medical students, the relevant literature has not been synthesized appropriately as to its value and effectiveness. To systematically review the literature regarding the impact of teaching EBM to medical students on their EBM knowledge, attitudes, skills and behaviors. MEDLINE, SCOPUS, Web of science, ERIC, CINAHL and Current Controlled Trials up to May 2011 were searched; backward and forward reference checking of included and relevant studies was also carried out. Two investigators independently extracted data and assessed the quality of the studies. 10,111 potential studies were initially found, of which 27 were included in the review. Six studies examined the effect of clinically integrated methods, of which five had a low quality and the other one used no validated assessment tool. Twelve studies evaluated the effects of seminars, workshops and short courses, of which 11 had a low quality and the other one lacked a validated assessment tool. Six studies examined e-learning, of which five having a high or acceptable quality reported e-learning to be as effective as traditional teaching in improving knowledge, attitudes and skills. One robust study found problem-based learning less effective compared to usual teaching. Two studies with high or moderate quality linked multicomponent interventions to improved knowledge and attitudes. No included study assessed the long-term effects of the teaching of EBM. Our findings indicated that some EBM teaching strategies have the potential to improve knowledge, attitudes and skills in undergraduate medical students, but the evidenced base does not demonstrate superiority of one method. There is no evidence demonstrating transfer to clinical practice.


Microdevice in Cellular Pathology: Microfluidic Platforms for Fluorescence in situ Hybridization and Analysis of Circulating Tumor Cells.
Microfluidic devices enable the miniaturization, integration, automation, and parallelization of chemical and biochemical processes. This new technology also provides opportunity for expansion in the field of cellular pathology. Fluorescence in situ hybridization (FISH) is a well-known gene-based method to image genetic abnormalities. Development of a FISH microfluidic platform has offered the possibility of automation with significant time and cost reductions, which overcomes many drawbacks of the current protocols. Microfluidic devices are also powerful tools for single-cell analysis. Capturing the circulating tumor cells (CTCs) from blood samples is one of the most promising approaches to enable the early diagnosis of cancer. The microfluidic devices are also useful to isolate rare CTCs at high efficiency and purity. In this review, I outline recent FISH and CTC analyses using microfluidic devices, and describe their applications for the cellular diagnosis of cancers.


Treg and Th17 cells in inflammatory periapical disease: a systematic review.
The process involved in periapical lesions, which occur as an outcome of pulpal necrosis, is regulated by the immune system including regulatory T cells (Treg) and T helper 17 cell (Th17) responses. The objective of this study was to conduct a frequency systematic review to determine the presence of Treg/Th17 responses and the influence of these cells in the progression of chronic inflammatory periapical lesions in humans. A systematic computerized search was carried out in Pubmed, Medline, Web of Science and Scopus electronic databases from their date of inception through the first week of May 2017. In addition, the reference lists of the included articles and the grey literature were hand-searched. Articles that evaluated the presence and influence of Treg/Th17 in the progression of human periapical lesions were included. Study selection and the quality assessment of the included articles (using the Newcastle-Ottawa scale) were carried out by two authors. Fifty-seven titles/abstracts were screened and eight studies met the eligibility criteria and were included in this systematic review. The included studies showed large variation in the type of periapical lesion assessed, mean age, age range, type of experiment and findings regarding the participation of Th17 and Treg in the status of inflammatory periapical lesions. The studies showed the involvement of Treg in the modulation of the inflammatory response in radicular cysts and periapical granulomas. This systematic review highlights the relationship between Treg and Th17 acting in a subtle balance inhibiting or promoting the progression of human periapical lesions.


Costs of formal and informal care in the last year of life for patients in receipt of specialist palliative care.
Economic evaluation of palliative care has been slow to develop and the evidence base remains small. This article estimates formal and informal care costs in the last year of life for a sample of patients who received specialist palliative care in three different areas in Ireland. Formal care costs are calculated for community, specialist palliative care, acute hospital and other services. Where possible, a bottom-up approach is used, multiplying service utilisation by unit cost. Informal care is valued at the replacement cost of care. Data on utilisation were collected during 215 'after death' telephone interviews with a person centrally involved in the care in the last year of life of decedents who received specialist palliative care in three areas in Ireland with varying levels of specialist palliative care. Mean total formal and informal costs in the last year of life do not vary significantly across the three areas. The components of formal costs, however, do vary across areas, particularly for hospital and specialist palliative care in the last 3 months of life. Costs in the last year of life for patients in receipt of specialist palliative care are considerable. Where inpatient hospice care is available, there are potential savings in hospital costs to offset specialist palliative care inpatient costs. Informal care accounts for a high proportion of costs during the last year of life in each area, underlining the important role of informal caregivers in palliative care.


Interventions for preventing unintended pregnancies among adolescents.
Unintended pregnancy among adolescents represents an important public health challenge in high-income countries, as well as middle- and low-income countries. Numerous prevention strategies such as health education, skills-building and improving accessibility to contraceptives have been employed by countries across the world, in an effort to address this problem. However, there is uncertainty regarding the effects of these interventions, hence the need to review the evidence-base. To assess the effects of primary prevention interventions (school-based, community/home-based, clinic-based, and faith-based) on unintended pregnancies among adolescents. We searched all relevant studies regardless of language or publication status up to November 2015. We searched the Cochrane Fertility Regulation Group Specialised trial register, The Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2015 Issue 11), MEDLINE, EMBASE, LILACS, Social Science Citation Index and Science Citation Index, Dissertations Abstracts Online, The Gray Literature Network, HealthStar, PsycINFO, CINAHL and POPLINE and the reference lists of articles. We included both individual and cluster randomised controlled trials (RCTs) evaluating any interventions that aimed to increase knowledge and attitudes relating to risk of unintended pregnancies, promote delay in the initiation of sexual intercourse and encourage consistent use of birth control methods to reduce unintended pregnancies in adolescents aged 10 years to 19 years. Two authors independently assessed trial eligibility and risk of bias, and extracted data. Where appropriate, binary outcomes were pooled using a random-effects model with a 95% confidence interval (Cl). Where appropriate, we combined data in meta-analyses and assessed the quality of the evidence using the GRADE approach. We included 53 RCTs that enrolled 105,368 adolescents. Participants were ethnically diverse. Eighteen studies randomised individuals, 32 randomised clusters (schools (20), classrooms (6), and communities/neighbourhoods (6). Three studies were mixed (individually and cluster randomised). The length of follow up varied from three months to seven years with more than 12 months being the most common duration. Four trials were conducted in low- and middle- income countries, and all others were conducted in high-income countries. Multiple interventionsResults showed that multiple interventions (combination of educational and contraceptive-promoting interventions) lowered the risk of unintended pregnancy among adolescents significantly (RR 0.66, 95% CI 0.50 to 0.87; 4 individual RCTs, 1905 participants, moderate quality evidence. However, this reduction was not statistically significant from cluster RCTs. Evidence on the possible effects of interventions on secondary outcomes (initiation of sexual intercourse, use of birth control methods, abortion, childbirth, sexually transmitted diseases) was not conclusive.Methodological strengths included a relatively large sample size and statistical control for baseline differences, while limitations included lack of biological outcomes, possible self-report bias, analysis neglecting clustered randomisation and the use of different statistical tests in reporting outcomes. Educational interventionsEducational interventions were unlikely to significantly delay the initiation of sexual intercourse among adolescents compared to controls (RR 0.95, 95% CI 0.71 to 1.27; 2 studies, 672 participants, low quality evidence).Educational interventions significantly increased reported condom use at last sex in adolescents compared to controls who did not receive the intervention (RR 1.18, 95% CI 1.06 to 1.32; 2 studies, 1431 participants, moderate quality evidence).However, it is not clear if the educational interventions had any effect on unintended pregnancy as this was not reported by any of the included studies. Contraceptive-promoting interventionsFor adolescents who received contraceptive-promoting interventions, there was little or no difference in the risk of unintended first pregnancy compared to controls (RR 1.01, 95% CI 0.81 to 1.26; 2 studies, 3,440 participants, moderate quality evidence).The use of hormonal contraceptives was significantly higher in adolescents in the intervention group compared to those in the control group (RR 2.22, 95% CI 1.07 to 4.62; 2 studies, 3,091 participants, high quality evidence) A combination of educational and contraceptive-promoting interventions appears to reduce unintended pregnancy among adolescents. Evidence for programme effects on biological measures is limited. The variability in study populations, interventions and outcomes of included trials, and the paucity of studies directly comparing different interventions preclude a definitive conclusion regarding which type of intervention is most effective.


Effects of pretreatment with reducing sugars or an enzymatic cocktail before extrusion of fava bean on nitrogen metabolism and performance of dairy cows.
The aim of this study was to determine the effects of pretreatment with reducing sugars or with an enzymatic cocktail before extrusion of fava bean on intake, milk yield and composition, N partitioning, and plasma and ruminal parameters. The main hypothesis was that these pretreatment conditions would allow better N protection in the rumen compared with classic pretreatment before extrusion, thanks to an increase of sugar proportion, either exogenous or endogenous with enzymatic actions. Sixteen Holstein cows were used in a 4 × 4 Latin square design experiment. Cow were fed a diet with a 56:44 forage to concentrate ratio and containing 16.2% of crude protein (CP; dry matter basis). Concentrate consisted of fava bean:linseed blends (90:10%, raw basis) distributed either raw, extruded after pretreatment without additive, extruded after pretreatment with reducing sugars, or extruded after pretreatment with an enzymatic cocktail. The experimental blends provided 53% of total CP in the diet. Intake, milk yield and composition, ruminal pH, volatile fatty acids and ammonia kinetics, apparent total-tract nutrient digestibilities and N partitioning, Maillard compounds in feed and feces, plasma AA, and 15N natural enrichment were measured. Data were analyzed using analysis of variance according to the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Extrusion without additive during pretreatment led to higher contents of Maillard compounds in the blend, a lower enzymatic CP degradability, a numerically lower ammonia content in ruminal fluid, and a trend of higher plasma EAA concentration, all suggesting a decrease in degradability of proteins in the rumen, and a subsequent increase in metabolizable protein supply. Compared with pretreatment without additive, adding reducing sugars or an enzymatic cocktail during pretreatment led to an increase in Maillard compound contents in the extruded blends, and to an increase (+50 mg/L) in rumen ammonia content. With reducing sugars, digestibility of Nɛ-carboxymethyl-lysine was numerically lower, and plasma EAA concentration tended to decrease, suggesting an overprotection of proteins in the intestine. With the enzymatic cocktail, plasma EAA concentration was similar than without additive during pretreatment, suggesting a similar metabolizable protein supply with both treatments. Finally, no change in N partitioning between milk, urine, and feces was observed whatever the diet.


A female head-neck model for rear impact simulations.
Several mathematical cervical models of the 50th percentile male have been developed and used for impact biomechanics research. However, for the 50th percentile female no similar modelling efforts have been made, despite females being subject to a higher risk of soft tissue neck injuries. This is a limitation for the development of automotive protective systems addressing Whiplash Associated Disorders (WADs), most commonly caused in rear impacts, as the risk for females sustaining WAD symptoms is double that of males. In this study, a finite element head and neck model of a 50th percentile female was validated in rear impacts. A previously validated ligamentous cervical spine model was complemented with a rigid body head, soft tissues and muscles. In both physiological flexion-extension motions and simulated rear impacts, the kinematic response at segment level was comparable to that of human subjects. Evaluation of ligament stress levels in simulations with varied initial cervical curvature revealed that if an individual assumes a more lordotic posture than the neutral, a higher risk of WAD might occur in rear impact. The female head and neck model, together with a kinematical whole body model which is under development, addresses a need for tools for assessment of automotive protection systems for the group which is at the highest risk to sustain WAD.


Power Doppler scanning in the diagnosis of carotid body tumors.
The aim of this work was to show contribution of power Doppler imaging in the diagnosis of the carotid body tumors. Six patients with a nontender mass beneath the mandibular angle were evaluated with gray scale and power Doppler sonography. Well-defined, solid, weakly hyperechoic masses were noted on gray scale sonography in the carotid bifurcation. Power Doppler sonography showed abundant flow, characterized as an intense blush, throughout the entire tumor in all patients. We believe that invasive and expensive diagnostic modalities are not necessary to evaluate carotid body tumors. Gray scale sonography and power Doppler imaging are sufficient for primary diagnosis of carotid body tumors.


Marginal Discrepancy of Cast Copings to Abutments with Three Different Luting Agents.
The purpose of this study was to evaluate the MD (marginal discrepancy) on the calcinable copings in abutments for cemented prostheses with three luting agents. Sixty-four analogs of CeraOne-type abutments (NACONIH code, Titanium Fix Implant Sytem SP, Brazil) were divided into four groups (n = 16). The copings were cast and placed on the CeraOne abutment analogs and cemented with eugenol-free zinc oxide (EfZO) (n = 16), with glass ionomer (GI) (n = 16), and with zinc phosphate (ZP) (n = 16), and as a control group, there were CeraOne plastic copings (NACOC code, Titanium Fix Implant Sytem SP, Brazil) (n = 16) which were not cemented with any material. After 24 hours, the MD of the four groups was measured. MD was evaluated using a stereoscopic microscope (Leica EZ4 W, Leica Microsystems, Germany) with an increase of ×100. MD was measured at four predetermined and equidistant sites with respect to the marginal line of the cast adaptation. The measurement was made from the distance between the free edge of the cast cylinder and the margin of preparation of the titanium abutment, with a level of statistical significance of p < 0.05. Of the three fixing agents, the ZP was found to have the highest MD (53.59 ± 14.21 μm); however, the lowest MD (41.72 ± 9.10 μm) was found in the GI group. These differences are statistically significant at p < 0.001. In summary, according to our results, it was found that ZP cement showed the highest MD after cementation, followed by the glass ionomer, while EfZO showed the lowest MD.


Dividing attention between two transparent motion surfaces results in a failure of selective attention.
In object-based attention, it is easier to divide attention between features within a single object than between features across objects. In this study we test the prediction of several capacity models in order to best characterize the cost to dividing attention between objects. Here we studied behavioral performance on a divided attention task in which subjects attended to the motion and luminance of overlapping random dot kinemategrams, specifically red upward moving dots superimposed with green downward moving dots. Subjects were required to detect brief changes (transients) in the motion or luminance within the same surface or across different surfaces. There were two primary results. First, the dual-task deficit was large when attention was divided across two surfaces and near zero when attention was divided within a surface. This is consistent with limited-capacity processing across surfaces and unlimited-capacity processing within a surface-a pattern predicted by established theories of object-based attention. Second and unexpectedly, there was evidence of crosstalk between features: when cued to monitor transients on one surface, response rates were inflated by the presence of a transient on the other surface. Such crosstalk is a failure of selective attention between surfaces.


Pulmonary sarcoidosis.
Sarcoidosis is a granulomatous disease of unknown cause, occurs worldwide and has a highly variable prevalence. The disease is typically dominant in the lungs, although it can affect virtually any organ and is unpredictable in its clinical course. The severity of pulmonary sarcoidosis ranges from incidentally discovered radiographic abnormalities in asymptomatic patients to a chronic progressive disease that is refractory to treatment. Mortality from sarcoidosis appears to have increased in the past three decades, with respiratory failure being the most common cause of sarcoidosis-related death. Pulmonary fibrosis, extensive disease on high-resolution chest CT, impaired lung function, and pulmonary hypertension are well established predictors of poor clinical outcomes. In patients who need systemic therapy to control their disease, corticosteroids are the most commonly used first-line treatment, with antimetabolites generally representing an alternative for patients who are unresponsive to corticosteroids or who cannot tolerate them. Indeed, corticosteroid therapy is associated with toxic effects that correlate with both the cumulative dose and duration of treatment. The scarcity of truly effective therapies and shortage of reliable predictors of the unpredictable development of disease in individual patients greatly contribute to making sarcoidosis such a difficult disease to manage.


Effect of Antidepressant Switching vs Augmentation on Remission Among Patients With Major Depressive Disorder Unresponsive to Antidepressant Treatment: The VAST-D Randomized Clinical Trial.
Less than one-third of patients with major depressive disorder (MDD) achieve remission with their first antidepressant. To determine the relative effectiveness and safety of 3 common alternate treatments for MDD. From December 2012 to May 2015, 1522 patients at 35 US Veterans Health Administration medical centers who were diagnosed with nonpsychotic MDD, unresponsive to at least 1 antidepressant course meeting minimal standards for treatment dose and duration, participated in the study. Patients were randomly assigned (1:1:1) to 1 of 3 treatments and evaluated for up to 36 weeks. Switch to a different antidepressant, bupropion (switch group, n = 511); augment current treatment with bupropion (augment-bupropion group, n = 506); or augment with an atypical antipsychotic, aripiprazole (augment-aripiprazole group, n = 505) for 12 weeks (acute treatment phase) and up to 36 weeks for longer-term follow-up (continuation phase). The primary outcome was remission during the acute treatment phase (16-item Quick Inventory of Depressive Symptomatology-Clinician Rated [QIDS-C16] score ≤5 at 2 consecutive visits). Secondary outcomes included response (≥50% reduction in QIDS-C16 score or improvement on the Clinical Global Impression Improvement scale), relapse, and adverse effects. Among 1522 randomized patients (mean age, 54.4 years; men, 1296 [85.2%]), 1137 (74.7%) completed the acute treatment phase. Remission rates at 12 weeks were 22.3% (n = 114) for the switch group, 26.9% (n = 136)for the augment-bupropion group, and 28.9% (n = 146) for the augment-aripiprazole group. The augment-aripiprazole group exceeded the switch group in remission (relative risk [RR], 1.30 [95% CI, 1.05-1.60]; P = .02), but other remission comparisons were not significant. Response was greater for the augment-aripiprazole group (74.3%) than for either the switch group (62.4%; RR, 1.19 [95% CI, 1.09-1.29]) or the augment-bupropion group (65.6%; RR, 1.13 [95% CI, 1.04-1.23]). No significant treatment differences were observed for relapse. Anxiety was more frequent in the 2 bupropion groups (24.3% in the switch group [n = 124] vs 16.6% in the augment-aripiprazole group [n = 84]; and 22.5% in augment-bupropion group [n = 114]). Adverse effects more frequent in the augment-aripiprazole group included somnolence, akathisia, and weight gain. Among a predominantly male population with major depressive disorder unresponsive to antidepressant treatment, augmentation with aripiprazole resulted in a statistically significant but only modestly increased likelihood of remission during 12 weeks of treatment compared with switching to bupropion monotherapy. Given the small effect size and adverse effects associated with aripiprazole, further analysis including cost-effectiveness is needed to understand the net utility of this approach. clinicaltrials.gov Identifier: NCT01421342.


Bag exchange in continuous ambulatory peritoneal dialysis without use of a face mask: experience of five years.
This article describes our five-year experience of continuous ambulatory peritoneal dialysis (CAPD) with bag exchanges performed without use of a face mask. The study took place in the renal unit at a university hospital. All patients admitted to the CAPD program from February 1995 to March 2000 were trained to perform bag exchanges without use of a face mask. Occurrence of peritonitis episodes was the outcome of interest. We evaluated 94 patients (52 women, 42 men) with a mean age of 48 +/- 21 years and a total follow-up of 50,502 patient-days. During that time, 79 episodes of peritonitis occurred in 46 patients, for a peritonitis rate of 0.57 episodes/year. The most frequently isolated micro-organisms were Staphylococcus epidermidis in 20 patients (25.3%) and S. aureus in 11 patients (13.9%). Renal transplantation was the major cause of drop-out [23 patients (43.4%)], followed by peritonitis [14 patients (26.4%)], death due to cardiovascular complications [9 patients (17.0%)], loss of ultrafiltration [2 patients (3.8%)], and other causes [5 patients (9.4%)]. The probability of being free of peritonitis at 12 months was 0.60, and at 60 months, 0.37. Peritonitis rates during the study period were not different from those reported by other centers, supporting the hypothesis that routine use of a face mask during CAPD bag exchange may be unnecessary.


Variation in emergency department use of cervical spine radiography for alert, stable trauma patients.
To, assess the emergency department use of cervical spine radiography for alert, stable adult trauma patients in terms of utilization, yield for injury and variation in practices among hospitals and physicians. Retrospective survey of health records. Emergency departments of 6 teaching and 2 community hospitals in Ontario and British Columbia. Consecutive alert, stable adult trauma patients seen with potential cervical spine injury between July 1, 1994, and June 30, 1995. Total number of eligible patients, referral for cervical spine radiography (overall, by hospital and by physician), presence of cervical spine injury, patient characteristics and hospitals associated with use of radiography. Of 6855 eligible patients, cervical spine radiography was ordered for 3979 (58.0%). Only 60 (0.9%) patients were found to have an acute cervical spine injury (fracture, dislocation or ligamentous instability); 98.5% of the radiographic films were negative for any significant abnormality. The demographic and clinical characteristics of the patients were similar across the 8 hospitals, and no cervical spine injuries were missed. Significant variation was found among the 8 hospitals in the rate of ordering radiography (p < 0.0001), from a low of 37.0% to a high of 72.5%. After possible differences in case severity and patient characteristics at each hospital were controlled for, logistic regression analysis revealed that 6 of the hospitals were significantly associated with the use of radiography. At 7 hospitals, there was significant variation in the rate of ordering radiography among the attending emergency physicians (p < 0.05), from a low of 15.6% to a high of 91.5%. Despite considerable variation among institutions and individual physicians in the ordering of cervical spine radiography for alert, stable trauma patients with similar characteristics, no cervical spine injuries were missed. The number of radiographic films showing signs of abnormality was extremely low at all hospitals. The findings suggest that cervical spine radiography could be used more efficiently, possibly with the help of a clinical decision rule.


Integration, segregation, and binocular combination.
The human visual system can accurately judge the mean of a distribution of different orientation samples. We ask whether the site of this integration is before or after the sites of binocular combination and disparity processing. Furthermore, we are interested in whether the efficiency with which local orientation information is integrated depends on the eye of origin. Our results suggest that orientation integration occurs after binocular integration but before disparity coding. We show that the effectiveness of added orientation noise is not only less than expected on signal or noise grounds but also that it depends on the dominance of the eye to which it is presented, suggesting an interocular opponent interaction in which the dominant eye input has higher gain.


The nicotinic acetylcholine receptor Dalpha7 is required for an escape behavior in Drosophila.
Acetylcholine is the major excitatory neurotransmitter in the central nervous system of insects. Mutant analysis of the Dalpha7 nicotinic acetylcholine receptor (nAChR) of Drosophila shows that it is required for the giant fiber-mediated escape behavior. The Dalpha7 protein is enriched in the dendrites of the giant fiber, and electrophysiological analysis of the giant fiber circuit showed that sensory input to the giant fiber is disrupted, as is transmission at an identified cholinergic synapse between the peripherally synapsing interneuron and the dorsal lateral muscle motor neuron. Moreover, we found that gfA1, a mutation identified in a screen for giant fiber defects more than twenty years ago, is an allele of Dalpha7. Therefore, a combination of behavioral, electrophysiological, anatomical, and genetic data indicate an essential role for the Dalpha7 nAChR in giant fiber-mediated escape in Drosophila.


Sputum CD34+IL-5Ralpha+ cells increase after allergen: evidence for in situ eosinophilopoiesis.
Eosinophil lineage-committed progenitors increase in the bone marrow of subjects with asthma developing allergen-induced airway hyperresponsiveness and eosinophilia. Also, higher numbers of circulating eosinophil/basophil cfu have been demonstrated 24 hours after allergen inhalation and in bronchial and nasal biopsies of allergic individuals. These cells may undergo in situ eosinophilopoiesis, suggesting that after allergen inhalation, progenitor cells traffic from the bone marrow to the airways, providing an ongoing source of effector cells. To examine this possibility, CD34(+) and CD34(+)IL-5Ralpha(+) cells were measured in induced sputum from allergic subjects with asthma at baseline and at 7 and 24 hours after allergen and diluent inhalation, using flow cytometry. Isolated early responders (n = 9) were contrasted to dual responders (n = 9), who develop allergen-induced sputum and blood eosinophilia and airway hyperresponsiveness, and to normal control subjects. At baseline, there were significantly fewer sputum eosinophils and CD34(+) cells in normal control subjects compared with subjects with asthma. Sputum CD34(+) cells increased at 7 hours after allergen inhalation in both groups of subjects with asthma, which was sustained at 24 hours in the dual responder group only, associated with sustained increases in sputum CD34(+)IL-5Ralpha(+) cells, eosinophils, and interleukin-5. These results indicate that eosinophil progenitors can migrate to the airways and may differentiate toward an eosinophilic phenotype.


Bee venom effects on ubiquitin proteasome system in hSOD1(G85R)-expressing NSC34 motor neuron cells.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that results from a progressive loss of motor neurons. Familial ALS (fALS) is caused by missense mutations in Cu, Zn-superoxide dismutase 1 (SOD1) that frequently result in the accumulation of mutant protein aggregates that are associated with impairments in the ubiquitin-proteasome system (UPS). UPS impairment has been implicated in many neurological disorders. Bee venom (BV) extracted from honey bees has been used as a traditional medicine for treating inflammatory diseases and has been shown to attenuate the neuroinflammatory events that occur in a symptomatic ALS animal model. NSC34 cells were transiently transfected with a WT or G85R hSOD1-GFP construct for 24 hrs and then stimulated with 2.5 μg/ml BV for 24 hrs. To determine whether a SOD1 mutation affects UPS function in NSC34 cells, we examined proteasome activity and performed western blotting and immunofluorescence using specific antibodies, such as anti-misfolded SOD1, anti-ubiquitin, anti-GRP78, anti-LC3, and anti-ISG15 antibodies. We found that GFP-hSOD1G85R overexpression induced SOD1 inclusions and reduced proteasome activity compared with the overexpression of GFP alone in NSC34 motor neuronal cells. In addition, we also observed that BV treatment restored proteasome activity and reduced the accumulation of ubiquitinated and misfolded SOD1 in GFP-hSOD1G85R-overexpressing NSC34 motor neuronal cells. However, BV treatment did not activate the autophagic pathway in these cells. Our findings suggest that BV may rescue the impairment of the UPS in ALS models.


Effect of preoperative angiotensin-converting enzyme inhibitor and angiotensin II receptor blocker use on hemodynamic variables in pediatric patients undergoing cardiopulmonary bypass.
Some have suggested that children undergoing cardiac surgery who receive angiotensin-converting enzyme (ACE) inhibitors experience a greater degree of hypotension after anesthesia induction and in the immediate postcardiopulmonary bypass period than children who did not receive these drugs. Therefore, we examined the effect of ACE inhibitor/angiotensin II receptor blocker (ARB) therapy on intraoperative hemodynamics and vasopressor use in pediatric patients undergoing cardiac surgery. In a retrospective cohort study of patients younger than 18 years who underwent cardiopulmonary bypass between March 1, 2010, and April 1, 2011, we compared intraoperative hemodynamics and vasopressor use between patients who received preoperative ACE inhibitor/ARB therapy and those who did not. The primary outcome was vasoactive infusion score after cardiopulmonary bypass. The occurrence of hypotension did not differ significantly between the ACE inhibitor/ARB group and the control group during induction of anesthesia or at any time point after cardiopulmonary bypass. At 0, 30, 60, and 90 minutes after cessation of cardiopulmonary bypass, patients on ACE inhibitor/ARB therapy tended to have a higher vasoactive infusion score (7.1, 7.6, 9.4, and 11.3) than patients in the control group (6.3, 6.1, 6.0, and 6.7). Although this difference became more pronounced over time, it did not reach statistical significance. The use of preoperative ACE inhibitors and ARBs in pediatric patients undergoing cardiac surgery did not significantly increase the incidence of hypotension after induction of anesthesia and did not increase significantly the vasoconstrictor requirements upon weaning from cardiopulmonary bypass; however, additional prospective studies are needed.