Single incision laparoscopic hepatectomy: Advances in laparoscopic liver surgery.
Laparoscopic liver surgery is now an established practice in many institutions. It is a safe and feasible approach in experienced hands. Single incision laparoscopic surgery (SILS) has been performed for cholecystectomies, nephrectomies, splenectomies and obesity surgery. However, the use of SILS in liver surgery has been rarely reported. We report our initial experience in seven patients on single incision laparoscopic hepatectomy (SILH). From October 2010 to September 2012, seven patients underwent single-incision laparoscopic liver surgery. The abdomen was approached through a 25 mm periumbilical incision. No supplemental ports were required. The liver was transected using a combination of LigaSure™ (Covidien-Valleylab. Boulder. USA), Harmonic Scalpel and Ligaclips (Ethicon Endo-Surgery, Inc.). Liver resection was successfully completed for the seven patients. The procedures consisted of two partial resections of segment three, two partial resections of segment five and three partial resections of segment six. The mean operative time was 98.3 min (range: 60-150 min) and the mean estimated blood loss was 57 ml (range: 25-150 ml). The postoperative courses were uneventful and the mean hospital stay was 5.1 days (range: 1-13 days). Pathology identified three benign and four malignant liver tumours with clear margins. SILH is a technically feasible and safe approach for wedge resections of the liver without oncological compromise and with favourable cosmetic results. This surgical technique requires relatively advanced laparoscopic skills. Further studies are needed to determine the potential advantages of this technique, apart from the better cosmetic result, compared to the conventional laparoscopic approach.


Use of preventive dental care among medicaid-enrolled, school-aged US children in immigrant and nonimmigrant families: trends in Pennsylvania From 2005 through 2010.
We describe trends in receipt of preventive dental care among Medicaid-enrolled children in Pennsylvania between 2005 and 2010, comparing the US children of immigrants with their co-ethnic peers in nonimmigrant families. We analyzed Pennsylvania Medicaid claims, birth records, and census data for children born in Pennsylvania and enrolled in Medicaid for 10 or more months during any of the calendar years assessed. Receipt of preventive dental care was more likely among Latino children in immigrant families than among their peers in nonimmigrant families; also, it was more likely among White children in immigrant families than among their peers in nonimmigrant families. Rates of preventive dental care use among African American and Asian children in immigrant and nonimmigrant families were comparable. From 2005 to 2010, the percentage of Latino children in nonimmigrant families who received preventive dental care increased from 33% to 61%. Changes in other groups were significant but less dramatic. Receipt of preventive dental care has increased among Medicaid-enrolled children in Pennsylvania, with marked gains among Latino children. Within each racial/ethnic group, the children of immigrants were either more likely than or equally likely as children in nonimmigrant families to receive care.


Acute effects of propranolol and metoprolol on plasma concentrations of parathyroid hormone and calcitonin in uraemic patients.
Nine uraemic patients not yet on dialysis received IV 1 microgram/kg/min of propranolol for 85 min after a priming dose of 1 mg. Fifteen days later six of them received IV 1.2 microgram/kg/min of metoprolol after a priming dose of 1.2 mg. Plasma concentrations of PTH and calcitonin decreased significantly with propranolol but not with metoprolol. No change was observed with either drug as regards plasma concentration of total and ionised Ca and PO4. Heart rate was decreased similarly with both drugs. We conclude that (i) propranolol acutely suppresses PTH and Calcitonin secretion in uraemic patients. This warrants further studies to assess its long term effects on the secretion of these hormones and on renal osteodystrophy; (ii) the contrast between the significant effect of propranolol and the lack of effect with metoprolol supports the concept that PTH and CT secretion are moderated through specific beta 2 receptors.


Effect of retinyl acetate on transglutaminase 2 activity in carcinogen treated rat liver.
Transglutaminase 2 (TG2) has been implicated in wound healing, cellular differentiation, apoptosis and cell survival. TG2 activity increases following acute and chronic liver injury; however, the role of TG2 in tumors, is controversial. TG2 is a retinoid-inducible enzyme. We investigated the effects of retinyl acetate (RA) on the activity and levels of TG2 during the initiation and promotion stages of liver cancer. p-Dimethylaminoazobenzene (p-DAB) was used as initiator and 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) was used as promoter in our model of carcinogenesis. Rats were divided into four groups of 24: control, corn oil control, p-DAB + TCDD, and p-DAB + TCDD + RA. Six rats from each group were sacrificed at days 30, 60, 90 and 120. TG2 activity decreased in the p-DAB + TCDD treated group, but TG2 immunostaining scores did not change by days 90 and 120. Neither TG2 enzyme activity nor the immunostaining score of TG2 protein changed in the tissues of the p-DAB + TCDD + RA group by days 90 and 120. TG2 activity was not be ameliorated by RA during the initiation or promotion stages of carcinogen induced liver cancer.


Cloning of the koi herpesvirus genome as an infectious bacterial artificial chromosome demonstrates that disruption of the thymidine kinase locus induces partial attenuation in Cyprinus carpio koi.
Koi herpesvirus (KHV) is the causative agent of a lethal disease in koi and common carp. In the present study, we describe the cloning of the KHV genome as a stable and infectious bacterial artificial chromosome (BAC) clone that can be used to produce KHV recombinant strains. This goal was achieved by the insertion of a loxP-flanked BAC cassette into the thymidine kinase (TK) locus. This insertion led to a BAC plasmid that was stably maintained in bacteria and was able to regenerate virions when permissive cells were transfected with the plasmid. Reconstituted virions free of the BAC cassette but carrying a disrupted TK locus (the FL BAC-excised strain) were produced by the transfection of Cre recombinase-expressing cells with the BAC. Similarly, virions with a wild-type revertant TK sequence (the FL BAC revertant strain) were produced by the cotransfection of cells with the BAC and a DNA fragment encoding the wild-type TK sequence. Reconstituted recombinant viruses were compared to the wild-type parental virus in vitro and in vivo. The FL BAC revertant strain and the FL BAC-excised strain replicated comparably to the parental FL strain. The FL BAC revertant strain induced KHV infection in koi carp that was indistinguishable from that induced by the parental strain, while the FL BAC-excised strain exhibited a partially attenuated phenotype. Finally, the usefulness of the KHV BAC for recombination studies was demonstrated by the production of an ORF16-deleted strain by using prokaryotic recombination technology. The availability of the KHV BAC is an important advance that will allow the study of viral genes involved in KHV pathogenesis, as well as the production of attenuated recombinant candidate vaccines.


Sharing risk management: an implementation model for cardiovascular absolute risk assessment and management in Australian general practice.
Despite considerable work in developing and validating cardiovascular absolute risk (CVAR) algorithms, there has been less work on models for their implementation in assessment and management. The aim of our study was to develop a model for a joint approach to its implementation based on an exploration of views of patients, general practitioners (GPs) and key informants (KIs). We conducted six focus group (three with GPs and three with patients) and nine KI interviews in Sydney. Thematic analysis was used with comparison to highlight the similarities and differences in perspectives of participants. Conducting CVAR was seen as more acceptable for regular patients rather than new patients for whom GPs had to attract their interest and build rapport before doing so at the next visit. GPs' interest and patients' positive attitude in managing risk were important in implementing CVAR. Long consultations, good communication skills and having a trusting relationship helped overcome the barriers during the process. All the participants supported engaging patients to self-assess their risk before the consultation and sharing decision making with GPs during consultation. Involving practice staff to help with the patient self-assessment, follow-up and referral would be helpful in implementing CVAR assessment and management, but GPs, patients and practices may need more support for this to occur. Multiple strategies are required to promote the better use of CVAR in the extremely busy working environment of Australian general practice. An implementation model has been developed based on our findings and the Chronic Care Model. Further research needs to investigate the effectiveness of the proposed model.


Update on diagnosis of canine hypothyroidism.
Until a clinically useful canine TSH assay is available, a valid test of thyroid hypofunction should (1) take into consideration valid (or at least predictive) measurement of the free T4 concentration in order to factor out the binding effects of drugs and nonthyroidal illness, and (2) should assess thyroid functional reserve. A validated equilibrium dialysis measurement of free T4 and the TSH stimulation test, provide the most useful diagnostic combination of tests now available. A therapeutic trial with L-thyroxine, despite being relatively safe, should be reserved once other diagnoses have been ruled out, and should be reevaluated to assure a clinical response which can be attributable to the medication.


Maternal germline-specific genes in the Asian malaria mosquito Anopheles stephensi: characterization and application for disease control.
Anopheles stephensi is a principal vector of urban malaria on the Indian subcontinent and an emerging model for molecular and genetic studies of mosquito biology. To enhance our understanding of female mosquito reproduction, and to develop new tools for basic research and for genetic strategies to control mosquito-borne infectious diseases, we identified 79 genes that displayed previtellogenic germline-specific expression based on RNA-Seq data generated from 11 life stage-specific and sex-specific samples. Analysis of this gene set provided insights into the biology and evolution of female reproduction. Promoters from two of these candidates, vitellogenin receptor and nanos, were used in independent transgenic cassettes for the expression of artificial microRNAs against suspected mosquito maternal-effect genes, discontinuous actin hexagon and myd88. We show these promoters have early germline-specific expression and demonstrate 73% and 42% knockdown of myd88 and discontinuous actin hexagon mRNA in ovaries 48 hr after blood meal, respectively. Additionally, we demonstrate maternal-specific delivery of mRNA and protein to progeny embryos. We discuss the application of this system of maternal delivery of mRNA/miRNA/protein in research on mosquito reproduction and embryonic development, and for the development of a gene drive system based on maternal-effect dominant embryonic arrest.


Alternative promoter usage at the Notch1 locus supports ligand-independent signaling in T cell development and leukemogenesis.
Loss of the transcription factor Ikaros is correlated with Notch receptor activation in T cell acute lymphoblastic leukemia (T-ALL). However, the mechanism remains unknown. We identified promoters in Notch1 that drove the expression of Notch1 proteins in the absence of a ligand. Ikaros bound to both canonical and alternative Notch1 promoters and its loss increased permissive chromatin, facilitating recruitment of transcription regulators. At early stages of leukemogenesis, increased basal expression from the canonical and 5'-alternative promoters initiated a feedback loop, augmenting Notch1 signaling. Ikaros also repressed intragenic promoters for ligand-independent Notch1 proteins that are cryptic in wild-type cells, poised in preleukemic cells, and active in leukemic cells. Only ligand-independent Notch1 isoforms were required for Ikaros-mediated leukemogenesis. Notch1 alternative-promoter usage was observed during T cell development and T-ALL progression. Thus, a network of epigenetic and transcriptional regulators controls conventional and unconventional Notch signaling during normal development and leukemogenesis.


Phoenix Mars Mission--the thermal evolved gas analyzer.
The Phoenix spacecraft that was launched to Mars in August 2007 landed safely on the Martian northern arctic region on May 25, 2008. It carried six experiments to study the history of water on the planet and search for organic molecules in the icy subsurface Martian soil. The spacecraft is a lander with an arm and scoop designed to dig a trench though the top soil to reach an expected ice layer near the surface. One of the instruments on board is the thermal evolved gas analyzer (TEGA), which consists of two components, a set of eight very small ovens that will heat samples of the ice soil mixtures from the trench to release imbedded gases and mineral decomposition products, and a mass spectrometer that serves as the analysis tool for the evolved gases, and also for measurements of the composition and isotopic ratios of the gases that comprise the atmosphere of Mars. The mass spectrometer is a miniature magnetic sector instrument controlled by microprocessor-driven power supplies. One feature is the gas enrichment cell that will increase the partial pressures of the noble gases in an atmosphere sample by removing all the active gases, carbon dioxide, and nitrogen, to improve the accuracy of their isotopic ratio measurements.


Spontaneous giving and calculated greed.
Cooperation is central to human social behaviour. However, choosing to cooperate requires individuals to incur a personal cost to benefit others. Here we explore the cognitive basis of cooperative decision-making in humans using a dual-process framework. We ask whether people are predisposed towards selfishness, behaving cooperatively only through active self-control; or whether they are intuitively cooperative, with reflection and prospective reasoning favouring 'rational' self-interest. To investigate this issue, we perform ten studies using economic games. We find that across a range of experimental designs, subjects who reach their decisions more quickly are more cooperative. Furthermore, forcing subjects to decide quickly increases contributions, whereas instructing them to reflect and forcing them to decide slowly decreases contributions. Finally, an induction that primes subjects to trust their intuitions increases contributions compared with an induction that promotes greater reflection. To explain these results, we propose that cooperation is intuitive because cooperative heuristics are developed in daily life where cooperation is typically advantageous. We then validate predictions generated by this proposed mechanism. Our results provide convergent evidence that intuition supports cooperation in social dilemmas, and that reflection can undermine these cooperative impulses.


Selective lysis of erythrocytes infected with the trophozoite stage of Plasmodium falciparum by polyene macrolide antibiotics.
The continuous increase in strains of the human malaria parasite Plasmodium falciparum resistant to most front-line antimalarial compounds is reason for grave clinical concern. The search for new drugs led us to investigate a number of membrane active polyene macrolide antibiotics, such as amphotericin B, nystatin, filipin and natamycin. The interaction of these compounds with sterols in bilayer cell membranes can lead to cell damage and ultimately cell lysis. The malaria parasite modifies the host erythrocyte membrane by changing the protein and lipid composition and thus the infected cell could be a selective target for membrane active compounds. We found that erythrocytes infected with the trophozoite stage of P. falciparum were particularly susceptible to lysis by amphotericin B (Fungizone) and, to a lesser extent, nystatin, as determined by ELISA and various microscopy assays. Liposomal amphotericin B (AmBisome) displayed a similar specificity for parasitised erythrocytes, but complete lysis required a longer incubation period. In contrast, filipin and natamycin did not distinguish between normal and parasite-infected erythrocytes, but lysed both at similar concentrations. In addition, when added to ring-stage cultures, the amphotericin B preparations and nystatin produced a marked disruption in parasite morphology in less than 2 h without an accompanying permeabilisation of the infected host cell, suggesting a second plasmodicidal mode of action. The results imply that selected polyene macrolide antibiotics or their derivatives could find application in the treatment of severe malaria caused by of P. falciparum.


Differential effects of unipolar versus bipolar depression on episodic memory updating.
Episodic memories, when reactivated, can be modified or updated by new learning. Since such dynamic memory processes remain largely unexplored in psychiatric disorders, we examined the impact of depression on episodic memory updating. Unipolar and bipolar depression patients, and age/education matched controls, first learned a set of objects (List-1). Two days later, participants in all three groups were either reminded of the first learning session or not followed by the learning of a new set of objects (List-2). Forty-eight hours later, List-1 recall was impaired in unipolar and bipolar patients compared to control participants. Further, as expected, control participants who received a reminder spontaneously recalled items from List-2 during recall of List-1, indicative of an updated List-1 memory. Such spontaneous intrusions were also seen in the unipolar and bipolar patients that received the reminder, suggesting that memory updating was unaffected in these two patient groups despite impaired recall of List 1. Unexpectedly, we observed a trend towards higher intrusions, albeit statistically insignificant, not only in the reminder but also in the no-reminder subgroups of bipolar patients. We probed this further in a second cohort by testing recall of List-2, which was also impaired in both depression groups. Again bipolar patients showed intrusions, but this time in the reverse order from List-1 into List-2, independent of a reminder. Taken together, despite impaired recall, updating of episodic memories was intact and unidirectional in unipolar depression. In contrast, indiscriminate updating, as evidenced by bidirectional interference between episodic memories, was seen in bipolar depression. These findings reveal a novel distinction between unipolar versus bipolar depression using a reactivation-dependent memory updating paradigm.


Automatic versus contingent mechanisms of sensory-driven neural biasing and reflexive attention.
Recent studies have generated debate regarding whether reflexive attention mechanisms are triggered in a purely automatic stimulus-driven manner. Behavioral studies have found that a nonpredictive "cue" stimulus will speed manual responses to subsequent targets at the same location, but only if that cue is congruent with actively maintained top-down settings for target detection. When a cue is incongruent with top-down settings, response times are unaffected, and this has been taken as evidence that reflexive attention mechanisms were never engaged in those conditions. However, manual response times may mask effects on earlier stages of processing. Here, we used event-related potentials to investigate the interaction of bottom-up sensory-driven mechanisms and top-down control settings at multiple stages of processing in the brain. Our results dissociate sensory-driven mechanisms that automatically bias early stages of visual processing from later mechanisms that are contingent on top-down control. An early enhancement of target processing in the extrastriate visual cortex (i.e., the P1 component) was triggered by the appearance of a unique bright cue, regardless of top-down settings. The enhancement of visual processing was prolonged, however, when the cue was congruent with top-down settings. Later processing in posterior temporal-parietal regions (i.e., the ipsilateral invalid negativity) was triggered automatically when the cue consisted of the abrupt appearance of a single new object. However, in cases where more than a single object appeared during the cue display, this stage of processing was contingent on top-down control. These findings provide evidence that visual information processing is biased at multiple levels in the brain, and the results distinguish automatically triggered sensory-driven mechanisms from those that are contingent on top-down control settings.


Comprehensive classification of the plant non-specific lipid transfer protein superfamily towards its sequence-structure-function analysis.
Non-specific Lipid Transfer Proteins (nsLTPs) are widely distributed in the plant kingdom and constitute a superfamily of related proteins. Several hundreds of different nsLTP sequences-and counting-have been characterized so far, but their biological functions remain unclear. It has been clear for years that they present a certain interest for agronomic and nutritional issues. Deciphering their functions means collecting and analyzing a variety of data from gene sequence to protein structure, from cellular localization to the physiological role. As a huge and growing number of new protein sequences are available nowadays, extracting meaningful knowledge from sequence-structure-function relationships calls for the development of new tools and approaches. As nsLTPs show high evolutionary divergence, but a conserved common right handed superhelix structural fold, and as they are involved in a large number of key roles in plant development and defense, they are a stimulating case study for validating such an approach. In this study, we comprehensively investigated 797 nsLTP protein sequences, including a phylogenetic analysis on canonical protein sequences, three-dimensional structure modeling and functional annotation using several well-established bioinformatics programs. Additionally, two integrative methodologies using original tools were developed. The first was a new method for the detection of (i) conserved amino acid residues involved in structure stabilization and (ii) residues potentially involved in ligand interaction. The second was a structure-function classification based on the evolutionary trace display method using a new tree visualization interface. We also present a new tool for visualizing phylogenetic trees. Following this new protocol, an updated classification of the nsLTP superfamily was established and a new functional hypothesis for key residues is suggested. Lastly, this work allows a better representation of the diversity of plant nsLTPs in terms of sequence, structure and function.


D-myo-inositol 1,4,5-trisphosphate analogues modified at the 3-position inhibit phosphatidylinositol 3-kinase.
Several natural and unnatural inositol phosphates and analogues were analyzed for their ability to inhibit the in vitro phosphatidylinositol 3-kinase (PI 3-kinase) activity immunoprecipitated from a leukemic T cell line by a p85 monoclonal antibody. A 3-position ring-modified analogue of D-myo-inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), L-chiro-inositol 2,3,5-trisphosphate (L-chiro-Ins(2,3,5)P3) and its phosphorothioate analogue, L-chiro-inositol 2,3,5-trisphosphorothioate, as well as the analogue benzene 1,2,4-trisphosphate induced reversible inhibition of PI 3-kinase activity, which correlated with decreased Vmax but unchanged Km values for PI 3-kinase. Other inositol phosphates, including D- and L-Ins(1,4,5)P3, D-myo-inositol 1,3,4,5-tetrakisphosphate, the enantiomers of myo-inositol 1,3,4-trisphosphate, DL-myo-inositol 1,4,6-trisphosphate (DL-Ins(1,4,6)P3), and DL-scyllo-inositol 1,2,4-trisphosphate (DL-scyllo-Ins(1,2,4)P3), did not inhibit PI 3-kinase activity under identical conditions. L-chiro-Ins(2,3,5)P3 closely resembles Ins(1,4,5)P3 and D-Ins(1,4,6)P3 except for a difference in the orientation of a single hydroxyl group at either the equivalent 3-OH or 2-OH position of Ins(1,4,5)P3, respectively. Similarly, L-chiro-Ins(2,3,5)P3 resembles D-scyllo-Ins(1,2,4)P3, but has a different orientation of both the equivalent 3-OH and 2-OH positions. Since Ins(1,4,5)P3, DL-Ins(1,4,6)P3, and DL-scyllo-Ins(1,2,4)P3 did not inhibit PI 3-kinase activity, this suggests that the orientation of the two hydroxyl groups at the 2- and 3-positions plays a pivotal role in the inhibitory action of inositol phosphate analogues on PI 3-kinase activity. Thus, inositol phosphate analogues inter alia are shown for the first time to inhibit PI 3-kinase and may be useful tools for determining the function of PI 3-kinase and its substrate binding specificities.


RLIMS-P 2.0: A Generalizable Rule-Based Information Extraction System for Literature Mining of Protein Phosphorylation Information.
We introduce RLIMS-P version 2.0, an enhanced rule-based information extraction (IE) system for mining kinase, substrate, and phosphorylation site information from scientific literature. Consisting of natural language processing and IE modules, the system has integrated several new features, including the capability of processing full-text articles and generalizability towards different post-translational modifications (PTMs). To evaluate the system, sets of abstracts and full-text articles, containing a variety of textual expressions, were annotated. On the abstract corpus, the system achieved F-scores of 0.91, 0.92, and 0.95 for kinases, substrates, and sites, respectively. The corresponding scores on the full-text corpus were 0.88, 0.91, and 0.92. It was additionally evaluated on the corpus of the 2013 BioNLP-ST GE task, and achieved an F-score of 0.87 for the phosphorylation core task, improving upon the results previously reported on the corpus. Full-scale processing of all abstracts in MEDLINE and all articles in PubMed Central Open Access Subset has demonstrated scalability for mining rich information in literature, enabling its adoption for biocuration and for knowledge discovery. The new system is generalizable and it will be adapted to tackle other major PTM types. RLIMS-P 2.0 online system is available online (http://proteininformationresource.org/rlimsp/) and the developed corpora are available from iProLINK (http://proteininformationresource.org/iprolink/).


Situation analysis of cutaneous leishmaniasis in an endemic area, south of Iran.
To update current situation of the cutaneous leishmaniasis (CL) in Kazerun County, southwest of Iran and to analyse the epidemiological aspects of the disease during 2005-2015. Data on CL were obtained from the Health Center of Kazerun County, and then were analysed and mapped using SPSS and Arc GIS 10.3. A total of 700 cases of CL were recorded during the study period with an overall decreasing trend from 2005 to 2015. More than 60% of the patients were inhabitants of rural areas and males were infected more than females. Although there was not a significant difference between gender, job categories, residence and CL infection (P > 0.05), age groups were significantly different (P < 0.05). But there was no significant correlation between monthly cases of the disease with average temperature (P > 0.05). Most of the acute lesions were found to be present on the hand, leg and face, respectively. The average CL incidence in the study area was calculated as 24.9/100000 population. A hot spot for the disease was found in southern part of the area (P < 0.05). This study revealed that CL is present in Kazerun country. Thus, effective monitoring and sustained surveillance system is crucial in counteracting the disease, and if possible, to eliminate it.


DNAseq Workflow in a Diagnostic Context and an Example of a User Friendly Implementation.
Over recent years next generation sequencing (NGS) technologies evolved from costly tools used by very few, to a much more accessible and economically viable technology. Through this recently gained popularity, its use-cases expanded from research environments into clinical settings. But the technical know-how and infrastructure required to analyze the data remain an obstacle for a wider adoption of this technology, especially in smaller laboratories. We present GensearchNGS, a commercial DNAseq software suite distributed by Phenosystems SA. The focus of GensearchNGS is the optimal usage of already existing infrastructure, while keeping its use simple. This is achieved through the integration of existing tools in a comprehensive software environment, as well as custom algorithms developed with the restrictions of limited infrastructures in mind. This includes the possibility to connect multiple computers to speed up computing intensive parts of the analysis such as sequence alignments. We present a typical DNAseq workflow for NGS data analysis and the approach GensearchNGS takes to implement it. The presented workflow goes from raw data quality control to the final variant report. This includes features such as gene panels and the integration of online databases, like Ensembl for annotations or Cafe Variome for variant sharing.


Mechanism of antibody synthesis: size differences between mouse kappa chains.
Structural analysis of immunoglobulin light chains has been carried out in an attempt to elucidate the genetic mechanisms involved in antibody synthesis. Analysis of two mouse kappa-chain proteins is almost complete. The differences are localized in one-half of the molecules, and do not reflect the operation of any one mutational mechanism. The peculiar character of the differences is discussed with reference to various theories of antibody formation. The finding that the two proteins differ in size is incompatible with certain proposed theories.


PH domain of ELMO functions in trans to regulate Rac activation via Dock180.
The members of the Dock180 superfamily of proteins are novel guanine nucleotide exchange factors (GEF) for Rho family GTPases and are linked to multiple biological processes from worms to mammals. ELMO is a critical regulator of Dock180, and the Dock180-ELMO complex functions as a bipartite GEF for Rac. We identified a mechanism wherein the PH domain of ELMO, by binding the Dock180-Rac complex in trans, stabilizes Rac in the nucleotide-free transition state. Mutagenesis studies reveal that this ELMO PH domain-dependent regulation is essential for the Dock180-ELMO complex to function in phagocytosis and cell migration. Genetic rescue studies in Caenorhabditis elegans using ELMO and its homolog CED-12 support the above observations in vivo. These data reveal a new mode of action of PH domains and a novel, evolutionarily conserved mechanism by which a bipartite GEF can activate Rac.


High-Molecular-Weight Polynucleotides by Transferase-Catalyzed Living Chain-Growth Polycondensation.
We present terminal deoxynucleotidyl transferase-catalyzed enzymatic polymerization (TcEP) for the template-free synthesis of high-molecular-weight, single-stranded DNA (ssDNA) and demonstrate that it proceeds by a living chain-growth polycondensation mechanism. We show that the molecular weight of the reaction products is nearly monodisperse, and can be manipulated by the feed ratio of nucleotide (monomer) to oligonucleotide (initiator), as typically observed for living polymerization reactions. Understanding the synthesis mechanism and the reaction kinetics enables the rational, template-free synthesis of ssDNA that can be used for a range of biomedical and nanotechnology applications.


Identification and characterisation of a prototype for a new class of competitive PPARγ antagonists.
Understanding of the physiological role of peroxisome proliferator-activated receptor gamma (PPARγ) offers new opportunities for the treatment of cancers, immune disorders and inflammatory diseases. In contrast to PPARγ agonists, few PPARγ antagonists have been studied, though they do exert immunomodulatory effects. Currently, no therapeutically useful PPARγ antagonist is commercially available. The aim of this study was to identify and kinetically characterise a new competitive PPARγ antagonist for therapeutic use. A PPARγ-dependent transactivation assay was used to kinetically characterise (E)-2-(5-((4-methoxy-2-(trifluoromethyl)quinolin-6-yl)methoxy)-2-((4-(trifluoromethyl)benzyl)oxy)-benzylidene)-hexanoic acid (MTTB) in kidney, T and monocytic cell lines. Cytotoxic effects were analysed and intracellular accumulation of MTTB was assessed by tandem mass spectrometry (LC-MS/MS). Potential interactions of MTTB with the PPARγ protein were suggested by molecular docking analysis. In contrast to non-competitive, irreversible inhibition caused by 2-chloro-5-nitrobenzanilide (GW9662), MTTB exhibited competitive antagonism against rosiglitazone in HEK293T and Jurkat T cells, with IC50 values in HEK293T cells of 4.3µM and 1.6µM, using the PPARγ ligand binding domain (PPARγ-LBD) and the full PPARγ protein, respectively. In all cell lines used, however, MTTB showed much higher intracellular accumulation than GW9662. MTTB alone exhibited weak partial agonistic effects and low cytotoxicity. Molecular docking of MTTB with the PPARγ-LBD supported direct interaction with the nuclear receptor. MTTB is a promising prototype for a new class of competitive PPARγ antagonists. It has weak partial agonistic and clear competitive antagonistic characteristics associated with rapid cellular uptake. Compared to commercially available PPARγ modulators, this offers the possibility of dose regulation of PPARγ and immune responses.


Topiramate use in pregnancy and the birth prevalence of oral clefts.
First marketed in the USA in 1996, topiramate (TPM) is an antiepileptic drug later approved for migraine prophylaxis, and in 2012 for weight loss in combination with phentermine. Some studies indicate an elevated prevalence of oral cleft (OC) in infants exposed to TPM in utero. We evaluated the association between TPM use in early pregnancy and the risk of OC. This retrospective cohort study used 1997-2011 automated data from four sources: HealthCore and OptumInsight (commercial insurance claims), Truven Health (Medicaid claims), and Kaiser Permanente Northern California Region (electronic medical records). We compared the prevalence of OCs in infants of women exposed to TPM in the first trimester (TPM cohort) with the prevalence in infants of women formerly exposed to TPM or other antiepileptic drugs (formerly exposed [FE] cohort) and infants of women with similar medical profiles (SMPs) to the TPM cohort that were not exposed to TPM (SMP cohort). To control for confounding, we used stratification and standardization for individual variables and propensity score deciles. The birth prevalence of OCs was 0.36% (7/1945) in the TPM cohort, 0.14% (20/13 512) in the FE cohort, and 0.07% (9/13 614) in the SMP cohort. Standardized by site, the prevalence ratio (PR) for TPM versus FE was 2.5 (95% CI: 1.0-6.0) and for TPM versus SMP was 5.4 (95% CI: 2.0-14.6). Adjustment for covariates one at a time or by propensity score yielded similar results. Consistent with other recent epidemiologic research, first-trimester TPM exposure was associated with an elevated birth prevalence of OC.


Understanding CPR architecture: an HIM professional's guide.
The continued evolution of the CPR offers numerous career opportunities for HIM professionals. But a knowledge and understanding of the fundamental concepts is critical for those who would take advantage of these opportunities. The authors offer an overview of the differences and interaction between the CPR component parts, the basic clinical and business functions they support, and the technology used to implement the CPR.


Transcranial magnetic stimulation of medial-frontal cortex impairs the processing of angry facial expressions.
Growing evidence suggests that the recognition of different emotional states involves at least partly separable neural circuits. Here we assessed the discrimination of both anger and happiness in healthy subjects receiving transcranial magnetic stimulation (TMS) over the medial-frontal cortex or over a control site (mid-line parietal cortex). We found that TMS over the medial-frontal cortex impairs the processing of angry, but not happy, facial expressions of emotion.


Personal tobacco pack display before and after the introduction of plain packaging with larger pictorial health warnings in Australia: an observational study of outdoor café strips.
We tested whether prevalence of cigarette pack display and smoking at outdoor venues and pack orientation changed following the introduction of plain packaging and larger pictorial health warnings in Australia. Between October and April 2011-12 (pre-plain packaging, pre-PP) and 2012-13 (post-plain packaging, post-PP), we counted patrons, smokers and tobacco packs at cafés, restaurants and bars with outdoor seating. Pack type (fully branded, plain or unknown) and orientation were noted. Rates of pack display, smoking and pack orientation were analysed using multi-level Poisson regression. Pack display declined by 15% [adjusted incident rate ratio (IRR) = 0.85, 95% confidence interval (CI) = 0.79-0.91, P < 0.001], driven by a 23% decline in active smoking (IRR = 0.77, 95% CI = 0.71-0.84, P < 0.001) between phases. The decline in pack display coincided with the full implementation of plain packaging from December 2012, was stronger in venues with children present and was limited to mid and high socio-economic status (SES) areas. The proportion of packs orientated face-up declined from 85.4% of fully branded packs pre-PP to 73.6% of plain packs post-PP (IRR = 0.87, 95% CI = 0.79-0.95, P = 0.002). Alternatively, the proportions concealed by telephones, wallets or other items (4.4% of fully branded packs pre-PP and 9.5% of plain packs post-PP; IRR = 2.33, 95% CI = 1.72-3.17, P < 0.001) and in an external case (1.5-3.5% of all packs; IRR = 2.79, 95% CI = 1.77-4.40, P < 0.001) increased. Low SES areas evidenced the greatest increase in pack concealment and the greatest decline in face-up pack orientation. Following Australia's 2012 policy of plain packaging and larger pictorial health warnings on cigarette and tobacco packs, smoking in outdoor areas of cafés, restaurants and bars and personal pack display (packs clearly visible on tables) declined. Further, a small proportion of smokers took steps to conceal packs that would otherwise be visible. Both are promising outcomes to minimize exposure to tobacco promotion.


Bone density and fracture risk following SBRT for non-spine bone metastases.
This retrospective study evaluated local recurrence (LR) and fracture risk in non-spine bone metastases treated with SBRT. 181 lesions in 116 patients are reported. The median dose was 27 Gy (range 15-40) in 3 fractions (range 1-6). The cumulative incidence of LR was 2.8%, 7.2% and 12.5% at 6 mo, 1 yr and 2 yrs. Fractures occurred in 11 lesions (6%). Radioresistant histology and increasing PTV predicted for LR on univariate analysis, while rib location was associated with control. Increasing PTV remained a significant predictor for LR on multivariate analysis. Univariate predictors of fracture risk included female gender, lytic lesions and poorer KPS. Average CT-approximated L1 trabecular attenuation in patients with fracture was significantly lower than in patients without fracture (112.2 vs. 142.6 Hounsfield units). In the largest series to date, we report excellent local control for SBRT to non-spine bone metastases and a novel relationship between CT-based bone quality assessment and fracture risk.


A possible antihypertensive mechanism of propranolol: antagonism of angiotensin II enhancement of sympathetic nerve transmission through prostaglandins.
The effects of propranolol on angiotensin II (AII) enhancement of sympathetic nerve transmission were investigated in the in situ blood-perfused mesenteric vascular bed of the rat. Angiotensin II in subpressor concentrations (3 ng/ml) potentiated the vasoconstrictor responses to both sympathetic nerve stimulation (NS) and exogenous norepinephrine (NE). The dl-propranolol had no effect on the basal vasoconstrictor responses to NS and NE, yet inhibited the AII-enhanced vasoconstrictor responses to NS by 47% (p less than 0.05) and 81% (p less than 0.001) at 100 and 300 ng/ml respectively. In contrast, the potentiation of NE responses by AII was unaffected by propranolol. A similar blockade of AII enhancement of NS was observed with the d-isomer of propranolol. Dibucaine (300 ng/ml), a local anesthetic, failed to alter the basal or AII-enhanced responses to either NS or NE. Indomethacin, a prostaglandin synthetase inhibitor (5 mg/kg, s.c.), abolished the inhibitory effect of dl-propranolol on AII enhancement of NS. Prostaglandin E2 (PGE2), but not prostaglandin I2, (3 ng/ml) inhibited AII enhancement of NS without altering the basal response to NS or NE in indomethacin-pretreated animals. Intraarterial infusions of dl-propranolol, d-propranolol, AII, and dl-propranolol-plus-AII into the superior mesenteric artery increased mesenteric venous PGE2 concentrations from 216 +/- 33 to 355 +/- 33 (p less than 0.01), 328 +/- 44 (p less than 0.05), 325 +/- 27 (p less than 0.02), and 407 +/- 44 pg/ml (p less than 0.01) respectively. We conclude that propranolol antagonizes AII enhancement of NS by increasing prostaglandin levels in vascular tissue. Furthermore, these findings suggest that propranolol may exert its antihypertensive effect through the release of prostaglandins when used in therapeutic doses in excess of those required for beta-adrenergic blockade.


Professional formation: extending medicine's lineage of service into the next century.
In his 1910 report on medical education, Flexner emphasized the importance of competency in basic sciences. Less widely recognized is that he also emphasized the necessity of liberal education. On the Flexner Report's 100th anniversary, medicine is challenged to realize Flexner's full vision for medical education to ensure that physicians are prepared to lead lives of compassion and service as well as to perform with technical proficiency. To meet the complex medical and social challenges of the next century, medical educators must continue to promote cognitive expertise while concurrently supporting "professional formation"-the moral and professional development of students, their ability to stay true to their personal service values and the core values of the profession, and the integration of their individual maturation with growth in clinical competency. The goal of professional formation is to anchor students to foundational principles while helping them navigate the inevitable moral conflicts in medical practice. The consequences of inadequate support for professional formation are profound, impacting individual learners, patients, the profession, and society at large. Among the many successful professional formation projects nationally, two long-standing programs are described in modest detail to identify common elements that might guide future developments elsewhere. Key elements include experiential and reflective processes, use of personal narratives, integration of self and expertise, and candid discussion within a safe community of learners. Committing to professional formation within medical education will require transformation of formal and informal curricula and will necessitate a rebalancing of attention and financial support within schools of medicine.


Effects of alkalosis on skeletal muscle metabolism and performance during exercise.
This study examined the effects of extracellular alkalosis on the metabolism and performance of perfused rat hindlimb muscles during electrical stimulation. Three acid-base conditions were used: control (C, normal acid-base state), metabolic alkalosis (MALK, increased bicarbonate concentration), and respiratory alkalosis (RALK, decreased PCO2). A one-pass system was used to perfuse the hindlimb via the femoral artery for 20 min at rest and during 5 min of tetanic stimulation via the sciatic nerve. The isometric tension generated by the gastrocnemius-plantaris-soleus muscle group was recorded. Arterial and venous perfusates were periodically sampled for substrate and metabolite measurements, and muscle samples were taken pre- and postperfusion. Peak isometric tensions in C, MALK, and RALK were similar: 3,367 +/- 107, 3,317 +/- 110, and 3,404 +/- 69 g, respectively. The rate of tension decay was also unaffected by alkalosis and represented 78 and 55% of the peak tension following 2 and 5 min of stimulation, respectively. Muscle O2 uptake, glycogen utilization, and total lactate (La-) production were similar following 5 min of stimulation in all conditions. However, alkalosis resulted in an enhanced La- release from working muscle (peak La- release: C, 15.5 +/- 1.1; MALK, 19.7 +/- 1.6; RALK, 18.3 +/- 2.2 mumol/min), and a 15-20% reduction in intramuscular La- accumulation. Alkalosis had no effect on muscle creatine phosphate and ATP concentrations. Thus, in the perfused rat hindlimb, alkalosis was not associated with changes in tetanic force or glycolysis, but La- release from the working muscle was enhanced by increased extracellular pH and bicarbonate.


Arthropathy-like findings and a carpal tunnel syndrome as the presenting features of Scheie syndrome: Three cases from the same family.
Gökay S, Kardaş F, Kendirci M, Sözeri B. Arthropathy-like findings and a carpal tunnel syndrome as the presenting features of Scheie syndrome: Three cases from the same family. Turk J Pediatr 2018; 60: 344-347. Mucopolysaccharidosis (MPS) type I is a rare autosomal recessive disease caused by a deficiency of the lysosomal enzyme α-L-iduronidase. MPS I is divided into three subtypes based on the severity of symptoms: Hurler, Hurler-Scheie, and Scheie syndrome (severe, intermediate, and mild forms, respectively). Musculoskeletal involvement may be the only presenting sign in the patients with Scheie syndrome. We have reviewed three cases with prominent features of carpal tunnel syndrome (CTS) at the onset of their disease. Diagnosis was delayed in almost all cases (range 16-19 years). During one year of follow-up period, alleviations of the pain in the hands of patients were observed after enzyme replacement therapy. MPS type I should be considered in the differential diagnosis of the patients with CTS in the first and second decades of life, particularly with stiffness of the fingers and difficulty using the hands without an inflammatory component. An increased awareness of the disease may contribute to more accurate diagnosis, and patients may benefit from early intervention.


Bridging and downstaging treatments for hepatocellular carcinoma in patients on the waiting list for liver transplantation.
Several therapeutic procedures have been proposed as bridging treatments for patients with hepatocellular carcinoma (HCC) awaiting liver transplantation (LT). The most used treatments include transarterial chemoembolization and radiofrequency ablation. Surgical resection has also been successfully used as a bridging procedure, and LT should be considered a rescue treatment in patients with previous HCC resection who experience tumor recurrence or post-treatment severe decompensation of liver function. The aims of bridging treatments include decreasing the waiting list dropout rate before transplantation, reducing HCC recurrence after transplantation, and improving post-transplant overall survival. To date, no data from prospective randomized studies are available; however, for HCC patients listed for LT within the Milan criteria, prolonging the waiting time over 6-12 mo is a risk factor for tumor spread. Bridging treatments are useful in containing tumor progression and decreasing dropout. Furthermore, the response to pre-LT treatments may represent a surrogate marker of tumor biological aggressiveness and could therefore be evaluated to prioritize HCC candidates for LT. Lastly, although a definitive conclusion can not be reached, the experiences reported to date suggest a positive impact of these treatments on both tumor recurrence and post-transplant patient survival. Advanced HCC may be downstaged to achieve and maintain the current conventional criteria for inclusion in the waiting list for LT. Recent studies have demonstrated that successfully downstaged patients can achieve a 5-year survival rate comparable to that of patients meeting the conventional criteria without requiring downstaging.


Gene transfer of suicide genes for the treatment of malignant gliomas: efficacy, limitations, and perspectives for a combined immunotherapy.
The potential of gene therapy strategies for malignant gliomas that are based on retroviral-mediated transfer of a "suicide gene" such as Herpes Simplex Virus-thymidine kinase HSV-tk and subsequent treatment by a prodrug (ganciclovir, for example), has been emphasized by the promising results obtained by several groups. However, further experimental data as well as preliminary clinical results indicate that the low efficiency of retroviral-mediated gene transfer in vivo as well as difficulties for the diffusion of the prodrug inside the tumour mass can limit the efficacy of this form of gene therapy. To achieve a more effective limitation of tumour growth other approaches may be combined with the "suicide gene" strategy and the enhancement of the immunological response to the tumour by cytokine gene transfer is prominent among these approaches. The authors' experiments in nude mice confirm the antineoplastic role of IL-4 and encourage testing the effects of the simultaneous transfer of IL-4 and HSV-tk genes in immunocompetent animals.


Role of sympathetic nerve activity in the generation of vascular nitric oxide in urethane-anesthetized rats.
The aim of the present study was to examine the involvement of the sympathetic nervous system in the generation or release of vascular nitric oxide. In urethane-anesthetized rats, the administration of the novel nitric oxide synthesis inhibitor L-N-nitro arginine (LNA) (0.02 mmol/kg i.v.) increased mean arterial pressure and renal, mesenteric, and hindquarter vascular resistances. The intravenous administration of L-arginine (60 mg/kg plus 12 mg/kg/min i.v.) produced small reductions in arterial pressure and vascular resistances and abolished the hemodynamic effects of LNA. Pretreatment with the ganglion blocking agent chlorisondamine lowered mean arterial pressure and vascular resistances, abolished the LNA-induced pressor and renal vasoconstrictor response, and attenuated the increases in mesenteric and hindquarter resistances. In contrast, the vasodilator hydralazine lowered mean arterial pressure and vascular resistances to levels equivalent to that of ganglionic blockade; however, the subsequent administration of LNA still produced significant increases in arterial pressure and regional vascular resistances. In ganglion-blocked rats in which pressure and vascular resistances were returned to normal levels by infusion of arginine vasopressin or phenylephrine, the pressor and vasoconstrictor effects of LNA were restored. However, phenylephrine was significantly more efficacious and markedly exaggerated the action of LNA. These results suggest that the sympathetic nervous system plays an important role in modulating the synthesis or release of vascular nitric oxide through the effects of 1) normal sympathetic discharge, 2) humoral activation of alpha-adrenergic receptors, and 3) vascular tone per se.


Children hospitalized with severe acute respiratory syndrome-related illness in Toronto.
An outbreak of severe acute respiratory syndrome (SARS) occurred in the greater Toronto area between February and June 2003. We describe the clinical, laboratory, and epidemiologic features of children who were admitted to the Hospital for Sick Children, Toronto, with a presumptive diagnosis of suspect or probable SARS. A prospective investigational study protocol was established for the management of children with a presumptive diagnosis of suspect or probable SARS. All were ultimately classified as having probable SARS, suspect SARS, or another cause on the basis of their epidemiologic exposure, clinical and radiologic features, and results of microbiologic investigations. Twenty-five children were included; 10 were classified as probable SARS and 5 were classified as suspect SARS, and in 10 another cause was identified. The exposure consisted of direct contact with at least 1 adult probable SARS case in 11 children, travel from a World Health Organization-designated affected area in Asia in 9 children, and presence in a Toronto area hospital in which secondary SARS spread had occurred in 5 children. The predominant clinical manifestations of probable cases were fever, cough, and rhinorrhea. With the exception of 1 teenager, none of the children developed respiratory distress or an oxygen requirement, and all made full recoveries. Mild focal alveolar infiltrates were the predominant chest radiograph abnormality. Lymphopenia; neutropenia; thrombocytopenia; and elevated alanine aminotransferase, aspartate aminotransferase, and creatine kinase were present in some cases. Nasopharyngeal swab specimens were negative for the SARS-associated coronavirus by an in-house reverse transcriptase-polymerase chain reaction in all 25 children. Our results indicate that SARS is a relatively mild and nonspecific respiratory illness in previously healthy young children. The presence of fever in conjunction with a SARS exposure history should prompt one to consider SARS as a possible diagnosis in children irrespective of the presence or absence of respiratory symptoms. Reverse-transcriptase polymerase chain reaction analysis of nasopharyngeal specimens seems to be of little utility for the diagnosis of SARS during the early symptomatic phase of this illness in young children.


Effects of emotion and emotional valence on the neural correlates of episodic memory search and elaboration.
Successful retrieval of an event includes an initial search phase in which the information is accessed and a subsequent elaboration phase in which an individual expands on event details. Traditionally, functional neuroimaging studies examining episodic memory retrieval either have not made a distinction between these two phases or have focused on the initial search process. The current study used an extended retrieval trial to compare the neural correlates of search and elaboration and to examine the effects of emotion on each phase. Before scanning, participants encoded positive, negative, and neutral images paired with neutral titles. After a 30-min delay, participants engaged in a scanned recognition task in which they viewed the neutral titles and indicated whether the title had been presented with an image during the study phase. Retrieval was divided into an initial memory search and a subsequent 5-sec elaboration phase. The current study identified neural differences between the search and elaboration phases, with search being associated with widespread bilateral activations across the entire cortex and elaboration primarily being associated with increased activity in the medial pFC. The emotionality of the retrieval target was more influential during search relative to elaboration. However, valence influenced when the effect of emotion was greatest, with search engaging many more regions for positive events than negative ones, but elaboration engaging the dorsomedial pFC more for negative events than positive events.


Unequal division in Saccharomyces cerevisiae and its implications for the control of cell division.
The budding yeast, Saccharomyces cerevisiae, was grown exponentially at different rates in the presence of growth rate-limiting concentrations of a protein synthesis inhibitor, cycloheximide. The volumes of the parent cell and the bud were determined as were the intervals of the cell cycle devoted to the unbudded and budded periods. We found that S. cerevisiae cells divide unequally. The daughter cell (the cell produced at division by the bud of the previous cycle) is smaller and has a longer subsequent cell cycle than the parent cell which produced it. During the budded period most of the volume increase occurs in the bud and very little in the parent cell, while during the unbudded period both the daughter and the parent cell increase significantly in volume. The length of the budded interval of the cell cycle varies little as a function of population doubling time; the unbudded interval of the parent cell varies moderately; and the unbudded interval for the daughter cell varies greatly (in the latter case an increase of 100 min in population doubling time results in an increase of 124 min in the daughter cell's unbudded interval). All of the increase in the unbudded period occurs in that interval of G1 that precedes the point of cell cycle arrest by the S. cerevisiae alpha-mating factor. These results are qualitatively consistent with and support the model for the coordination of growth and division (Johnston, G. C., J. R. Pringle, and L. H. Hartwell. 1977. Exp. Cell. Res. 105:79-98.) This model states that growth and not the events of the DNA division cycle are rate limiting for cellular proliferation and that the attainment of a critical cell size is a necessary prerequisite for the "start" event in the DNA-division cycle, the event that requires the cdc 28 gene product, is inhibited by mating factor and results in duplication of the spindle pole body.


Let it flow: Morpholino knockdown in zebrafish embryos reveals a pro-angiogenic effect of the metalloprotease meprin alpha2.
Meprin metalloproteases are thought to be involved in basic physiological functions such as cell proliferation and tissue differentiation. However, the specific functions of these enzymes are still ambiguous, although a variety of growth factors and structural proteins have been identified as meprin substrates. The discovery of meprins alpha(1), alpha(2) and beta in teleost fish provided the basis for uncovering their physiological functions by gene silencing in vivo. A Morpholino knockdown in zebrafish embryos targeting meprin alpha(1) and beta mRNA caused defects in general tissue differentiation. But meprin alpha(2) morphants were affected more specifically and showed severe failures in the formation of the vascular system provoking the hypothesis of a pro-angiogenic effect. The blood circulation was largely diminished resulting in erythrocyte accumulation. These phenotypes mimic a previously described VEGF-A morphant, revealing a possible role of meprin alpha in VEGF-A activation. Indeed, human recombinant meprin alpha processed the vascular endothelial growth factor-A (VEGF-A) specifically, revealing the same cleavage products detectable for VEGF from zebrafish whole lysate. Our results demonstrate that meprin metalloproteases are important for cell differentiation and proliferation already during embryogenesis, predominantly by the activation of growth factors. Thus, we conclude that meprins play a significant role in VEGF-A processing, subsequently regulating angiogenesis. Therefore, meprin alpha might be a new therapeutic target in cardiovascular diseases or in tumor growth inhibition.


Acute psychosocial stress-mediated changes in the expression and methylation of perforin in chronic fatigue syndrome.
Perforin (PRF1) is essential for immune surveillance and studies report decreased perforin in chronic fatigue syndrome (CFS), an illness potentially associated with stress and/or infection. We hypothesize that stress can influence regulation of PRF1 expression, and that this regulation will differ between CFS and non-fatigued (NF) controls. We used the Trier Social Stress Test (TSST) as a standardized acute psychosocial stress, and evaluated its effect on PRF1 expression and methylation in CFS (n = 34) compared with NF (n = 47) participants. During the TSST, natural killer (NK) cells increased significantly in both CFS (P = <0.0001) and NF subjects (P = <0.0001). Unlike previous reports, there was no significant difference in PRF1 expression at baseline or during TSST between CFS and NF. However, whole blood PRF1 expression increased 1.6 fold during the TSST in both CFS (P = 0.0003) and NF (P = <0.0001). Further, the peak response immediately following the TSST was lower in CFS compared with NF (P = 0.04). In addition, at 1.5 hours post TSST, PRF1 expression was elevated in CFS compared with NF (whole blood, P = 0.06; PBMC, P = 0.02). Methylation of seven CpG sites in the methylation sensitive region of the PRF1 promoter ranged from 38%-79% with no significant differences between CFS and NF. Although, the average baseline methylation of all seven CpG sites did not differ between CFS and NF groups, it showed a significant negative correlation with PRF1 expression at all TSST time points in both CFS (r = -0.56, P = <0.0001) and NF (r = -0.38, P = <0.0001). Among participants with high average methylation (≥65%), PRF1 expression was significantly lower in CFS than NF subjects immediately following TSST. These findings suggest methylation could be an important epigenetic determinant of inter-individual differences in PRF1 expression and that the differences in PRF1 expression and methylation between CFS and NF in the acute stress response require further investigation.


A comparison of primary care graduates from schools with increasing production of family physicians to those from schools with decreasing production.
This study investigated factors related to declining interest in family medicine by US medical school graduates. A questionnaire was sent to all physicians who graduated from 24 medical schools in 1997-1999, and who entered a family medicine residency, and a randomly selected equal number of graduates from the same years who entered other primary care specialties. Between 1997 and 1999, 12 of these schools had increases and 12 had decreases in the proportion of graduates choosing family medicine residencies. Between 1997 and 1999, at schools with increasing proportions of graduates choosing family medicine, there were significant increases in the proportion of graduates who (1). had entered medical school with a specialty preference of family medicine, (2). spent their required family medicine clerkship at two or more sites, (3). ranked the competence of family medicine faculty highly, (4). reported the faculty member they most wanted to be like was a family physician, and (5). experienced clinical rotations in both family medicine and primary care. At schools with declines in the proportion of graduates choosing family medicine, there were significant declines in the proportion of graduates who (1). ranked the competence of family medicine faculty highly, (2). stated that they were encouraged to go into family medicine, and (3). reported that the faculty member they most wanted to be like was a family physician. In schools with decreases in family medicine graduates, there was a significant increase in the proportion of graduates intending a large city or suburban practice. Using binary logistic regression, the variables that remained significantly correlated with attending a school with increases or decreases in students selecting family medicine were the number of required clinical rotations in family medicine and primary care, the perception of the clinical competence of the family medicine faculty, and an intent to practice, or subsequently having a practice, in a rural area. Schools that want to increase their production of family physicians should consider admissions policies that select students inclined toward family medicine and rural practice, should adopt a curriculum that maximizes clinical training with family physicians and other primary care physicians, and should ensure that their family medicine faculty are perceived as competent role models.


Interresponse time changes as a function of water deprivation and amphetamine.
Drug effects on operant behavior are often characterized by their effects on rate of responding, usually expressed as the number of responses per unit of time. The time between two consecutive responses constitutes an interresponse time (IRT), and this measure has been used also to characterize the effects of drugs on operant behavior. IRTs which occur during a session can be classified on a statistical basis as: 1) short-IRT, an IRT(s) of short duratio generated by high-frequency responses; 2) pause, an IRT of long duratio generated by low-frequency responses; and 3) post-reinforcement pause, an IRT which immediately follows reinforcement. This investigation used three schedules of water reinforcement (fixed-ratio 20, fixed-interval 90-seconds and variable-interval 20-seconds) to examine how these IRT classes are influenced by changes in water deprivation conditions or amphetamine administration. Base-line IRT distributions depended upon the schedule of reinforcement. Changes induced by doses of amphetamine or alterations in level of water deprivation were compared and contrasted. Short-IRTs that characterized fixed-ratio 20 performance were resistant to change with increasing doses of amphetamine, but were increased in duration with decreasing water deprivation. Animals responding on a fixed-interval 90-second schedule showed a decreased postreinforcement pause after amphetamine, but an increased postreinforcement pause after access to water. An additional experiment studied the combined effects of presession water consumption and d-amphetamine administration on variable interval performances. Making water available before the session lowered the amphetamine dose-response curve along the vertical axis, suggesting that amphetamine did not mimic satiation. In most cases the effect of amphetamine and changing levels of water deprivation were dissimilar in their effects on IRT distributions, suggesting that amphetamine does not exert its major action on behavior through its adipsic effect.


Relationship between bidet toilet use and haemorrhoids and urogenital infections: a 3-year follow-up web survey.
Given the growing use of electric bidet toilets in Japan and other countries, we assessed the relationship between bidet toilet use and haemorrhoids or urogenital infections. Data were collected using a web-based longitudinal survey. In total, 10 305 subjects randomly selected from panels of a Japanese website research company for the baseline survey in 2013 were asked about their frequency of bidet toilet use and receipt of a doctor's diagnosis or subjective symptom of haemorrhoids and urogenital infections. One- and three-year follow-up surveys were performed in 2014 and 2016, respectively, and information on newly diagnosed/experienced outcomes occurring during the follow-up period were collected. Cumulative incidence of haemorrhoids and urogenital infections was not significantly increased by habitual use of a bidet toilet. In men, more habitual users reported subjective symptoms of irritated skin around the anus, which were newly experienced during follow-up than non-habitual users (adjusted risk ratio 1.36 (95% confidence interval 1.06-1.75)). Further studies are needed to confirm this relationship. Several of the outcomes were significantly more prevalent in habitual users, but these results were probably explained by reverse causation.


Comparisons of Female and Egg Assays to Identify Rotylenchulus reniformis Resistance in Cotton.
More plants can be screened for reniform nematode resistance each year if the time involved can be shortened. In this study, the hypothesis that female counts are as efficient as egg counts in identifying resistant genotypes was tested. In two greenhouse experiments Gossypium genotypes which varied from resistant to susceptible to reniform nematode (Rotylenchulus reniformis) were compared to a susceptible control cultivar. Infested field soil served as the inoculum source for the first experiment, and vermiform stages extracted from greenhouse cultures were used to infest soil in the second experiment. Six replicates of each genotype were harvested 25 d after planting and swollen females were counted. The remaining plants were harvested 35 d after planting and eggs extracted from the roots were counted. Processing and counting times recorded in the first experiment were similar for both assessment methods, but 10 additional days were required for egg-based assessment. Contrast analyses showed that assessments based on females per gram of root were equivalent to assessments based on eggs per gram of root for the five genotypes tested in the first experiment and for an expanded set of 13 genotypes tested in the second experiment. The results indicated that either life stage can be used to screen for resistance.


Endoscopic treatment for distal malignant biliary obstruction.
Distal malignant biliary obstruction (MBO) leads to obstructive jaundice as a result of when the bile excretion from the liver is disturbed and induces hepatic failure and sepsis, which when complicated with cholangitis, it becomes necessary to perform drainage for the MBO. For biliary drainage, we can perform a surgical bypass operation, percutaneous transhepatic biliary drainage (PTBD), endoscopic biliary drainage (EBD) via duodenal papilla, or endoscopic ultrasound (EUS)-guided biliary drainage (EUS-BD), which is a transgastrointestinal biliary drainage. Although currently we usually perform EBD for distal MBO to begin with, the choice is different for biliary drainage in patients in whom EBD has failed in a preoperative case or an unresectable case. In other words, we choose PTBD for preoperative cases, and PTBD or EUS-BD according to the ability of the institution for their procedures when EBD has failed. It is desirable not to choose a plastic stent (PS) but a self-expandable metallic stent (SEMS), in particular for the unresectable cases of pancreatic cancer it is desirable not to choose an uncovered SEMS but a covered SEMS in EBD. Nevertheless, further examinations are expected to decide which, a covered or uncovered SEMS, we should choose in unresectable biliary tract cancer (BTC) and whether we should select PS, SEMS or ENBD in preoperative cases.


Right- and left-loop short shRNAs have distinct and unusual mechanisms of gene silencing.
Small hairpin RNAs (shRNAs) having duplex lengths of 25-29 bp are normally processed by Dicer into short interfering RNAs (siRNAs) before incorporation into the RNA-induced silencing complex (RISC). However, shRNAs of ≤ 19 bp [short shRNAs (sshRNAs)] are too short for Dicer to excise their loops, raising questions about their mechanism of action. sshRNAs are designated as L-type or R-type according to whether the loop is positioned 3' or 5' to the guide sequence, respectively. Using nucleotide modifications that inhibit RNA cleavage, we show that R- but not L-sshRNAs require loop cleavage for optimum activity. Passenger-arm slicing was found to be important for optimal functioning of L-sshRNAs but much less important for R-sshRNAs that have a cleavable loop. R-sshRNAs could be immunoprecipitated by antibodies to Argonaute-1 (Ago1); complexes with Ago1 contained both intact and loop-cleaved sshRNAs. In contrast, L-sshRNAs were immunoprecipitated with either Ago1 or Ago2 and were predominantly sliced in the passenger arm of the hairpin. However, 'pre-sliced' L-sshRNAs were inactive. We conclude that active L-sshRNAs depend on slicing of the passenger arm to facilitate opening of the duplex, whereas R-sshRNAs primarily act via loop cleavage to generate a 5'-phosphate at the 5'-end of the guide strand.


The far-lateral approach for foramen magnum meningiomas.
Foramen magnum meningiomas (FMMs) are slow growing, most often intradural and extramedullary tumors that pose significant challenges to the skull base neurosurgeon. The indolent clinical course of FMMs and their insidious onset of symptoms are important factors that contribute to delayed diagnosis and relative large size at the time of presentation. Symptoms are often produced by compression of surrounding structures (such as the medulla oblongata, upper cervical spinal cord, lower cranial nerves, and vertebral artery) within a critically confined space. Since the initial pathological description of a FMM in 1872, various surgical approaches have been described with the aim of achieving radical tumor resection. The surgical treatment of FMMs has evolved considerably over the last 4 decades due to the progress in microsurgical techniques and development of a multitude of skull base approaches. Posterior and posterolateral FMMs can be safely resected via a standard midline suboccipital approach. However, controversy still exits regarding the optimal management of anterior or anterolateral lesions. Independently of technical variations and the degree of bone removal, all modern surgical approaches to the lower clivus and anterior foramen magnum derive from the posterolateral (or far-lateral) craniotomy originally described by Roberto Heros and Bernard George. This paper is a review of the surgical management of FMMs, with emphasis on the far-lateral approach and its variations. Clinical presentation, imaging findings, important neuroanatomical correlations, recurrence rates, and outcomes are discussed.


I222 Neuraminidase mutations further reduce oseltamivir susceptibility of Indonesian Clade 2.1 highly pathogenic Avian Influenza A(H5N1) viruses.
We have tested the susceptibility to neuraminidase inhibitors of 155 clade 2.1 H5N1 viruses from Indonesia, isolated between 2006-2008 as well as 12 clade 1 isolates from Thailand and Cambodia from 2004-2007 using a fluorometric MUNANA-based enzyme inhibition assay. The Thailand and Cambodian clade 1 isolates tested here were all susceptible to oseltamivir and zanamivir, and sequence comparison indicated that reduced oseltamivir susceptibility we observed previously with clade 1 Cambodian isolates correlated with an S246G neuraminidase mutation. Eight Indonesian viruses (5%), all bearing I222 neuraminidase mutations, were identified as mild to extreme outliers for oseltamivir based on statistical analysis by box plots. IC50s were from 50 to 500-fold higher than the reference clade 1 virus from Viet Nam, ranging from 43-75 nM for I222T/V mutants and from 268-349 nM for I222M mutants. All eight viruses were from different geographic locales; all I222M variants were from central Sumatra. None of the H5N1 isolates tested demonstrated reduced susceptibility to zanamivir (IC50s all <5 nM). All I222 mutants showed loss of slow binding specifically for oseltamivir in an IC50 kinetics assay. We identified four other Indonesian isolates with higher IC50s which also demonstrated loss of slow binding, including one virus with an I117V mutation. There was a minimal effect on the binding of zanamivir and peramivir for all isolates tested. As H5N1 remains a potential pandemic threat, the incidence of mutations conferring reduced oseltamivir susceptibility is concerning and emphasizes the need for greater surveillance of drug susceptibility.


Twenty-four-week clevudine therapy showed potent and sustained antiviral activity in HBeAg-positive chronic hepatitis B.
Clevudine is a pyrimidine analogue with potent and sustained antiviral activity against HBV. The present study evaluated the safety and efficacy of 30 mg clevudine once daily for 24 weeks and assessed the durable antiviral response for 24 weeks after cessation of dosing. A total of 243 hepatitis B e antigen (HBeAg)-positive chronic hepatitis B patients were randomized (3:1) to receive clevudine 30 mg once daily (n=182) or placebo (n=61) for 24 weeks. Patients were followed for a further 24 weeks off therapy. Median serum HBV DNA reductions from baseline at week 24 were 5.10 and 0.27 log10 copies/mL in the clevudine and placebo groups, respectively (P<0.0001). Viral suppression in the clevudine group was sustained off therapy, with 3.73 log10 reduction at week 34 and 2.02 log10 reduction at week 48. At week 24, 59.0% of patients in the clevudine group had undetectable serum HBV DNA levels by Amplicor PCR assay (less than 300 copies/mL). The proportion of patients who achieved normalization of alanine aminotransferase (ALT) levels was 68.2% in the clevudine group and 17.5% in the placebo group at week 24 (P<0.0001). ALT normalization in the clevudine group was well maintained during post-treatment follow-up period. The incidence of adverse events (AEs) was similar between the clevudine group and the placebo group. No resistance to clevudine was detected with 24 weeks of administration of drug. A 24-week clevudine therapy was well tolerated and showed potent and sustained antiviral effect without evidence of viral resistance during treatment period in HBeAg-positive chronic hepatitis B.


Permissive role of neutrophils in pathogenesis of indomethacin-induced gastric lesions in rats.
We examined the possible role of neutrophils in the pathogenesis of indomethacin-induced gastric lesions, in comparison with prostaglandin (PG) deficiency. Rats were given indomethacin (35 mg/kg, s.c.) and killed 4 hr later. Gastric motility, mucosal PGE2 levels, and myeloperoxidase (MPO) activity were measured following indomethacin. Atropine was given s.c. 30 min before administration of indomethacin, while 16, 16-dimethyl PGE2 (dmPGE2) or anti-rat neutrophil antiserum (ANS) was given i.v. 10 min or 1 hr, respectively, before indomethacin treatment. Indomethacin reduced PGE2 contents in the stomach and produced hemorrhagic lesions in the stomach, with an increase of gastric motility and MPO activity. Indomethacin-induced gastric lesions were significantly prevented by dmPGE2 as well as atropine, at any time points during a 4 hr-test period. By contrast, the pretreatment of ANS did not prevent the development of gastric lesions when examined at either 1, 2 or 3 hr following indomethacin, but significantly reduced the severity of these lesions at 4 hr after indomethacin treatment. Both dmPGE2 and atropine inhibited the increase of gastric motility and MPO activity in response to indomethacin, whereas ANS prevented the increase of MPO activity, without any effect on the gastric hypermotility. These results confirmed that indomethacin-induced gastric lesions occurred in association with gastric hypermotility, in both atropine and PG-sensitive manners, and further suggest that the neutrophil activation/migration is not sufficient by itself to induce damage in the stomach and may be implicated in the process of later extension of damage.


Analysis of data from the International Outcome Inventory for Hearing Aids (IOI-HA) using Bayesian Item Response Theory.
IOI-HA response data are conventionally analysed assuming that the ordinal responses have interval-scale properties. This study critically considers this assumption and compares the conventional approach with a method using Item Response Theory (IRT). A Bayesian IRT analysis model was implemented and applied to several IOI-HA data sets. Anonymised IOI-HA responses from 13273 adult users of one or two hearing aids in 11 data sets using the Australian English, Dutch, German and Swedish versions of the IOI-HA. The raw ordinal responses to IOI-HA items do not represent values on interval scales. Using the conventional rating sum as an overall score introduces a scale error corresponding to about 10 - 15% of the true standard deviation in the population. Some interesting and statistically credible differences were demonstrated among the included data sets. It is questionable to apply conventional statistical measures like mean, variance, t-tests, etc., on the raw IOI-HA ratings. It is recommended to apply only nonparametric statistical test methods for comparisons of IOI-HA results between groups. The scale error can sometimes cause incorrect conclusions when individual results are compared. The IRT approach is recommended for analysis of individual results.


Catastrophic cognitions about coronavirus: the Oxford psychological investigation of coronavirus questionnaire [TOPIC-Q].
Cognitive therapies are developed on the principle that specific cognitive appraisals are key determinants in the development and maintenance of mental health disorders. It is likely that particular appraisals of the coronavirus pandemic will have explanatory power for subsequent mental health outcomes in the general public. To enable testing of this hypothesis we developed a questionnaire assessing coronavirus-related cognitions. 12 285 participants completed online a 46-item pool of cognitions about coronavirus and six measures of different mental health problems. The sample was randomly split into derivation and validation samples. Exploratory factor analyses determined the factor structure, selection of items, and model fit in the derivation sample. Confirmatory factor analysis (CFA) then tested this model in the validation sample. Associations of the questionnaire with mental health outcomes were examined. The 26-item, seven-factor, Oxford Psychological Investigation of Coronavirus Questionnaire [TOPIC-Q] was developed. CFA demonstrated a good model fit (χ2 = 2108.43, df = 278, p < 0.001, comparative fit index (CFI) = 0.950, Tucker-Lewis index (TLI) = 0.942, root mean square error of approximation (RMSEA) = 0.033, standardized root mean square residual (SRMR) = 0.038). The factors were: cognitions about (1) safety and vulnerability, (2) negative long-term impact, (3) having the virus, (4) spreading the virus, (5) social judgment, (6) negative self, and (7) being targeted. The questionnaire explained significant variance in depression (45.8%), social anxiety (37.3%), agoraphobia (23.2%), paranoia (27.3%), post-traumatic stress disorder (57.1%), and panic disorder (31.4%). Cognitions about negative long-term impact had the greatest explanatory power across disorders. TOPIC-Q provides a method to assess appraisals of the pandemic, which is likely to prove helpful both in longitudinal studies assessing mental health outcomes and in delivery of psychological therapy.


Medial ulnar collateral ligament reconstruction of the elbow in major league baseball players: Where do we stand?
The ulnar collateral ligament (UCL) is a vital structure to the overhead athlete, especially the baseball pitcher. For reasons not completely understood, UCL injuries have become increasingly more common in major league baseball (MLB) pitchers over the past 10 years. UCL reconstruction (UCLR) is the current gold standard of treatment for these injuries in MLB pitchers who wish to return to sport (RTS) at a high level and who have failed a course of non-operative treatment. Results following UCLR in MLB pitchers have been encouraging, with multiple RTS rates now cited at greater than 80%. Unfortunately, with the rising number of UCLR, there has also been a spike in the number of revision UCLR in MLB pitchers. Similar to primary UCLR, the etiology of the increase in revision UCLR, aside from an increase in the number of pitchers who have undergone a primary UCLR, remains elusive. The current literature has attempted to address several questions including those surrounding surgical technique (method of exposure, graft choice, management of the ulnar nerve, concomitant elbow arthroscopy, etc.), post-operative rehabilitation strategies, and timing of RTS following UCLR. While some questions have been answered, many remain unknown. The literature surrounding UCLR in MLB pitchers will be reviewed, and future directions regarding this injury in these high level athletes will be discussed.


Estriol retards and stabilizes atherosclerosis through an NO-mediated system.
Estriol (E3) has little effect on the female genitals. E3 is used in hormone replacement therapy, particularly in Europe and Japan, since it obviates the need for progestin administration. However, the effect of E3 on atherosclerosis has not been elucidated. In this study, we evaluated the effect of E3 on the progression of atherosclerosis in a rabbit model. Thirty-six rabbits total were used. Twenty-eight were bilaterally oophorectomized, and 8 were not. The rabbits were divided into 5 groups and treated for 12 weeks as follows. Gp I (n = 8) was fed a high cholesterol diet (HCD; standard diet plus 0.5% cholesterol); Gp II (n = 8) was fed a HCD with E3 (0.3 mg/kg/day); Gp III (n = 8) was fed a HCD with 17beta estradiol (E2) (0.1 mg/kg/day); Gp IV (n = 8), the non oophorectomized group, was fed a HCD; and Gp NC was oophorectomized (n = 4), and fed a regular diet. E3 treatment increased the plasma E2 and E3 levels in Gp II. The plasma lipid levels were not altered by the E2 or E3 treatment. A HCD diminished the acetylcholine-induced NO mediated relaxation in the thoracic aorta. The E2 treatment (Gp III) and E3 treatment (Gp II) restored the aortic basal NO release and the aortic cyclic GMP levels, particularly effectively in the E3 group. E3 treatment also decreased the atherosclerotic area, and its effect was comparable with E2 (surface involvement: 41.2 +/- 5.1% in Gp I; 10.1 +/- 2.7% in Gp II; and 6.5 +/- 1.3% in Gp III). All four hyperlipidemic groups showed an increase of eNOS mRNA in the aortae, and this was especially pronounced in Gps II and III. The level of peroxynitrite, as determined by immunohistochemical nitrotyrosine staining, was lower in Gps II and III than in Gp I. E3 strongly activates NO-mediated systems, and could play a role in retarding the progression of atherosclerosis and in stabilizing atheroma.


Targeted genotyping of variable number tandem repeats with adVNTR.
Whole-genome sequencing is increasingly used to identify Mendelian variants in clinical pipelines. These pipelines focus on single-nucleotide variants (SNVs) and also structural variants, while ignoring more complex repeat sequence variants. Here, we consider the problem of genotyping Variable Number Tandem Repeats (VNTRs), composed of inexact tandem duplications of short (6-100 bp) repeating units. VNTRs span 3% of the human genome, are frequently present in coding regions, and have been implicated in multiple Mendelian disorders. Although existing tools recognize VNTR carrying sequence, genotyping VNTRs (determining repeat unit count and sequence variation) from whole-genome sequencing reads remains challenging. We describe a method, adVNTR, that uses hidden Markov models to model each VNTR, count repeat units, and detect sequence variation. adVNTR models can be developed for short-read (Illumina) and single-molecule (Pacific Biosciences [PacBio]) whole-genome and whole-exome sequencing, and show good results on multiple simulated and real data sets.


Physical activity, physical fitness, and coronary heart disease risk factors.
The relationships between physical activity, physical fitness, and coronary heart disease risk factors measured in a large community sample were evaluated. Self-reported physical activity using a single question, maximal oxygen consumption estimates derived from the Pawtucket Heart Health Step Test, blood pressure, nonfasting lipids, and body mass index were cross-sectionally evaluated in 381 men and 556 women. The correlation of estimated maximal oxygen consumption and self-reported physical activity was modest but statistically significant (r = 0.13 in men and r = 0.19 in women). Blood pressure, body mass index, and HDL cholesterol were correlated with physical fitness (r = 0.24-0.65) and correlated to self-reported physical activity (r = 0.09-0.14). Evaluation of coronary heart disease risk factors using both physical activity and physical fitness revealed a complex relationship that generally showed a stronger relationship with measures of physical fitness than with physical activity. This study suggests that simultaneous measurement of physical activity and physical fitness may be useful in epidemiologic studies of habitual physical activity and chronic disease.


An update on pediatric bronchiectasis.
The prevalence and awareness of bronchiectasis not related to cystic fibrosis (CF) is increasing and it is now recognized as a major cause of respiratory morbidity, mortality and healthcare utilization worldwide. The need to elucidate the early origins of bronchiectasis is increasingly appreciated and has been identified as an important research priority. Current treatments for pediatric bronchiectasis are limited to antimicrobials, airway clearance techniques and vaccination. Several new drugs targeting airway inflammation are currently in development. Areas covered: Current management of pediatric bronchiectasis, including discussion on therapeutics, non-pharmacological interventions and preventative and surveillance strategies are covered in this review. We describe selected adult and pediatric data on bronchiectasis treatments and briefly discuss emerging therapeutics in the field. Expert commentary: Despite the burden of disease, the number of studies evaluating potential treatments for bronchiectasis in children is extremely low and substantially disproportionate to that for CF. Research into the interactions between early life respiratory tract infections and the developing immune system in children is likely to reveal risk factors for bronchiectasis development and inform future preventative and therapeutic strategies. Tailoring interventions to childhood bronchiectasis is imperative to halt the disease in its origins and improve adult outcomes.


Content Validity Index and Reliability of a New Protocol for Evaluation of Lifting Technique in the Powerlifting Squat and Deadlift.
Sjöberg, H, Aasa, U, Rosengren, M, and Berglund, L. Content validity index and reliability of a new protocol for evaluation of lifting technique in the powerlifting squat and deadlift. J Strength Cond Res 34(9): 2528-2536, 2020-The aim of the study was to create a protocol to cover aspects of technique considered to be associated with risk of injury in the powerlifting squat and deadlift and to examine the content validity and reliability of the aspects included in the protocols. For the content validity investigation, a consensus group of 3 powerlifting physiotherapists identified the domains of content (risk of injury) for 2 protocols (1 for squat and 1 for deadlift) of essential aspects of lifting technique through discussions and a review of the literature. Eight selected powerlifting experts rated the relevance of each aspect in relation to risk of injury (acute or by overuse), and a quantitative estimate of the content validity of each aspect was measured through calculations of a Content Validity Index (CVI). Aspects of low content validity were discarded, and the remainders were evaluated for their inter-rater and intra-rater reliability among 4 experienced powerlifters used to coaching and evaluating powerlifting technique. The reliability was calculated and analyzed with kappa and percentage of agreement. The final protocols included 17 aspects of squat technique and 10 aspects of deadlift technique that showed good to excellent CVI and percentage of agreement between 64 and 100%. The protocols, formed in this study, will provide evidence-based recommendations on safe lifting technique for coaches and strength practitioners' to use to make relevant assessments and instructions.


IL-17 is a neuromodulator of Caenorhabditis elegans sensory responses.
Interleukin-17 (IL-17) is a major pro-inflammatory cytokine: it mediates responses to pathogens or tissue damage, and drives autoimmune diseases. Little is known about its role in the nervous system. Here we show that IL-17 has neuromodulator-like properties in Caenorhabditis elegans. IL-17 can act directly on neurons to alter their response properties and contribution to behaviour. Using unbiased genetic screens, we delineate an IL-17 signalling pathway and show that it acts in the RMG hub interneurons. Disrupting IL-17 signalling reduces RMG responsiveness to input from oxygen sensors, and renders sustained escape from 21% oxygen transient and contingent on additional stimuli. Over-activating IL-17 receptors abnormally heightens responses to 21% oxygen in RMG neurons and whole animals. IL-17 deficiency can be bypassed by optogenetic stimulation of RMG. Inducing IL-17 expression in adults can rescue mutant defects within 6 h. These findings reveal a non-immunological role of IL-17 modulating circuit function and behaviour.


Investigation of temporal illicit drugs, alcohol and tobacco trends in Istanbul city: Wastewater analysis of 14 treatment plants.
Wastewater-based drug monitoring is a complementary tool that has been used worldwide in recent years, and many cities have periodically reported monitoring results. However, this study is the first to analyze drugs in wastewater in a single city with a high population during four periods simultaneously from 14 treatment plants. The aim was to estimate the consumption of conventional illicit drugs [amphetamine (AMP), methamphetamine (METH), 3,4-methylenedioxymethamphetamine (MDMA), cannabis, cocaine, and heroin], tobacco, and alcohol in 2019 for quarterly periods in Istanbul city, which has a population of almost 20 million, to aid in implementing evidence-based measures. Additionally, the seasonal variations among the 14 wastewater treatment plants and their weekday/weekend comparison of drug use patterns and consumption per substance were examined. Solid phase extraction was followed by liquid chromatography-tandem mass spectrometry and a previously validated method was enhanced by adding new parameters (morphine, cotinine, and ethyl sulfate), and satisfactory results were obtained. In this study, alcohol, nicotine, and cannabis had the highest estimated per capita consumption with mean values of 29655.3 mL/1000 inh/day, 5507.6 mg/1000 inh/day and 3607.0 mg/1000 inh/day, respectively. These results were followed by heroin and cocaine consumption with mean values of 557.0 and 200.9 mg/1000 inh/day, respectively, whereas AMP-type stimulants had the lowest values among the targeted substances. METH and cannabis were also highly consumed drugs when compared with the results of other metropolitan cities, whereas heroin consumption was remarkably high owing to Turkey's location on a possible heroin trafficking route. Because Istanbul is the business center of the country and has the potential to attract tourists in all four seasons, meaningful seasonal consumption differences were not observed for all substances.


A model for the self-organization of vesicular flux and protein distributions in the Golgi apparatus.
The generation of two non-identical membrane compartments via exchange of vesicles is considered to require two types of vesicles specified by distinct cytosolic coats that selectively recruit cargo, and two membrane-bound SNARE pairs that specify fusion and differ in their affinities for each type of vesicles. The mammalian Golgi complex is composed of 6-8 non-identical cisternae that undergo gradual maturation and replacement yet features only two SNARE pairs. We present a model that explains how distinct composition of Golgi cisternae can be generated with two and even a single SNARE pair and one vesicle coat. A decay of active SNARE concentration in aging cisternae provides the seed for a cis[Formula: see text]trans SNARE gradient that generates the predominantly retrograde vesicle flux which further enhances the gradient. This flux in turn yields the observed inhomogeneous steady-state distribution of Golgi enzymes, which compete with each other and with the SNAREs for incorporation into transport vesicles. We show analytically that the steady state SNARE concentration decays exponentially with the cisterna number. Numerical solutions of rate equations reproduce the experimentally observed SNARE gradients, overlapping enzyme peaks in cis, medial and trans and the reported change in vesicle nature across the Golgi: Vesicles originating from younger cisternae mostly contain Golgi enzymes and SNAREs enriched in these cisternae and extensively recycle through the Endoplasmic Reticulum (ER), while the other subpopulation of vesicles contains Golgi proteins prevalent in older cisternae and hardly reaches the ER.


Polymorphisms in oxidative stress and inflammation pathway genes, low-dose ionizing radiation, and the risk of breast cancer among US radiologic technologists.
Ionizing radiation, an established breast cancer risk factor, has been shown to induce oxidative damage and chronic inflammation. Polymorphic variation in oxidative stress and inflammatory-mediated pathway genes may modify radiation-related breast cancer risk. We estimated breast cancer risk for 28 common variants in 16 candidate genes involved in these pathways among 859 breast cancer cases and 1,083 controls nested within the US Radiologic Technologists cohort. We estimated associations between occupational and personal diagnostic radiation exposures with breast cancer by modeling the odds ratio (OR) as a linear function in logistic regression models and assessed heterogeneity of the dose-response across genotypes. There was suggestive evidence of an interaction between the rs5277 variant in PTGS2 and radiation-related breast cancer risk. The excess OR (EOR)/Gy from occupational radiation exposure = 5.5 (95%CI 1.2-12.5) for the GG genotype versus EOR/Gy < 0 (95%CI < 0-3.8) and EOR/Gy < 0 (95%CI < 0-14.8) for the GC and CC genotypes, respectively, (p (interaction) = 0.04). The association between radiation and breast cancer was not modified by other SNPs examined. This study suggests that variation in PTGS2 may modify the breast cancer risk from occupational radiation exposure, but replication in other populations is needed to confirm this result.