The Anal Pap Smear: Cytomorphology of squamous intraepithelial lesions.
BACKGROUND: Anal smears are increasingly being used as a screening test for anal squamous intraepithelial lesions (ASILs). This study was undertaken to assess the usefulness and limitations of anal smears in screening for ASILs. METHODS: The cytomorphological features of 200 consecutive anal smears collected in liquid medium from 198 patients were studied and findings were correlated with results of surgical biopsies and/or repeat smears that became available for 71 patients within six months. RESULTS: Adequate cellularity was defined as an average of 6 or more nucleated squamous cells/hpf. A glandular/transitional component was not required for adequacy. Dysplastic cells, atypical parakeratotic cells and bi/multinucleated cells were frequent findings in ASIL while koilocytes were infrequent. Smears from LSIL cases most frequently showed mildly dysplastic and bi/multinucleate squamous cells followed by parakeratotic cells (PK), atypical parakeratotic cells (APK), and koilocytes. HSIL smears contained squamous cells with features of moderate/severe dysplasia and many APKs. Features of LSIL were also found in most HSIL smears. CONCLUSIONS: In this study liquid based anal smears had a high sensitivity (98%) for detection of ASIL but a low specificity (50%) for predicting the severity of the abnormality in subsequent biopsy. Patients with cytologic diagnoses of ASC-US and LSIL had a significant risk (46-56%) of HSIL at biopsy. We suggest that all patients with a diagnosis of ASC-US and above be recommended for high resolution anoscopy with biopsy.


Executive Function, Dexterity, and Discharge Disposition in Older Intensive Care Unit Survivors.
Dexterity is a component of motor function. Executive function, a subdomain of cognition, may affect dexterity in older adults recovering from critical illness after discharge from an intensive care unit (ICU). To explore associations between executive function (attention and cognitive flexibility) and dexterity (fine motor coordination) in the early post-ICU period and examine dexterity by acuity of discharge disposition. The study involved 30 older adults who were functionally independent before hospitalization, underwent mechanical ventilation in the ICU, and had been discharged from the ICU 24 to 48 hours previously. Dexterity was evaluated with the National Institutes of Health Toolbox (NIHTB) Motor Battery 9-Hole Pegboard Dexterity Test (PDT); attention, with the NIHTB Cognition Battery Flanker Inhibitory Control and Attention Test (FICAT); and cognitive flexibility, with the NIHTB Cognition Battery Dimensional Change Card Sort Test (DCCST). Exploratory regression was used to examine associations between executive function and dexterity (fully corrected T scores). Independent-samples t tests were used to compare dexterity between participants discharged home and those discharged to a facility. FICAT (β = 0.375, P = .03) and DCCST (β = 0.698, P = .001) scores were independently and positively associated with PDT scores. Further, PDT scores were worse among participants discharged to a facility than among those discharged home (mean [SD], 26.71 [6.14] vs 36.33 [10.30]; t24 = 3.003; P = .006). Poor executive function is associated with worse dexterity; thus, dexterity may be a correlate of both post-ICU cognitive impairment and functional decline. Performance on dexterity tests could identify frail older ICU survivors at risk for worse discharge outcomes.


Regional expression of P2Y(4) receptors in the rat central nervous system.
P2Y receptors are G protein-coupled receptors composed of eight known subunits (P2Y(1, 2, 4, 6, 11, 12, 13, 14)), which are involved in different functions in neural tissue. The present study investigates the expression pattern of P2Y(4) receptors in the rat central nervous system (CNS) using immunohistochemistry and in situ hybridization. The specificity of the immunostaining has been verified by preabsorption, Western blot, and combined use of immunohistochemistry and in situ hybridization. Neurons expressing P2Y(4) receptors were distributed widely in the rat CNS. Heavy P2Y(4) receptor immunostaining was observed in the magnocellular neuroendocrine neurons of the hypothalamus, red nucleus, pontine nuclei, mesencephalic trigeminal nucleus, motor trigeminal nucleus, ambiguous nucleus, inferior olive, hypoglossal nucleus, and dorsal motor vagus nucleus. Both neurons and astrocytes express P2Y(4) receptors. P2Y(4) receptor immunostaining signals were mainly confined to cell bodies and dendrites of neurons, suggesting that P2Y(4) receptors are mainly involved in regulating postsynaptic events. In the hypothalamus, all the vasopressin (VP) and oxytocin (OT) neurons and all the orexin A neurons were immunoreactive for P2Y(4) receptors. All the neurons expressing P2Y(4) receptors were found to express N-methyl-D: -aspartate receptor 1 (NR1). These data suggest that purines and pyrimidines might be involved in regulation of the release of the neuropeptides VP, OT, and orexin in the rat hypothalamus via P2Y(4) receptors. Further, the physiological and pathophysiological functions of the neurons may operate through coupling between P2Y(4) receptors and NR1.


Gene expression polymorphism underpins evasion of host immunity in an asexual lineage of the Irish potato famine pathogen.
Outbreaks caused by asexual lineages of fungal and oomycete pathogens are a continuing threat to crops, wild animals and natural ecosystems (Fisher MC, Henk DA, Briggs CJ, Brownstein JS, Madoff LC, McCraw SL, Gurr SJ, Nature 484:186-194, 2012; Kupferschmidt K, Science 337:636-638, 2012). However, the mechanisms underlying genome evolution and phenotypic plasticity in asexual eukaryotic microbes remain poorly understood (Seidl MF, Thomma BP, BioEssays 36:335-345, 2014). Ever since the 19th century Irish famine, the oomycete Phytophthora infestans has caused recurrent outbreaks on potato and tomato crops that have been primarily caused by the successive rise and migration of pandemic asexual lineages (Goodwin SB, Cohen BA, Fry WE, Proc Natl Acad Sci USA 91:11591-11595, 1994; Yoshida K, Burbano HA, Krause J, Thines M, Weigel D, Kamoun S, PLoS Pathog 10:e1004028, 2014; Yoshida K, Schuenemann VJ, Cano LM, Pais M, Mishra B, Sharma R, Lanz C, Martin FN, Kamoun S, Krause J, et al. eLife 2:e00731, 2013; Cooke DEL, Cano LM, Raffaele S, Bain RA, Cooke LR, Etherington GJ, Deahl KL, Farrer RA, Gilroy EM, Goss EM, et al. PLoS Pathog 8:e1002940, 2012). However, the dynamics of genome evolution within these clonal lineages have not been determined. The objective of this study was to use a comparative genomics and transcriptomics approach to determine the molecular mechanisms that underpin phenotypic variation within a clonal lineage of P. infestans. Here, we reveal patterns of genomic and gene expression variation within a P. infestans asexual lineage by comparing strains belonging to the South American EC-1 clone that has dominated Andean populations since the 1990s (Yoshida K, Burbano HA, Krause J, Thines M, Weigel D, Kamoun S, PLoS Pathog 10e1004028, 2014; Yoshida K, Schuenemann VJ, Cano LM, Pais M, Mishra B, Sharma R, Lanz C, Martin FN, Kamoun S, Krause J, et al. eLife 2:e00731, 2013; Delgado RA, Monteros-Altamirano AR, Li Y, Visser RGF, van der Lee TAJ, Vosman B, Plant Pathol 62:1081-1088, 2013; Forbes GA, Escobar XC, Ayala CC, Revelo J, Ordonez ME, Fry BA, Doucett K, Fry WE, Phytopathology 87:375-380, 1997; Oyarzun PJ, Pozo A, Ordonez ME, Doucett K, Forbes GA, Phytopathology 88:265-271, 1998). We detected numerous examples of structural variation, nucleotide polymorphisms and loss of heterozygosity within the EC-1 clone. Remarkably, 17 genes are not expressed in one of the two EC-1 isolates despite apparent absence of sequence polymorphisms. Among these, silencing of an effector gene was associated with evasion of disease resistance conferred by a potato immune receptor. Our findings highlight the molecular changes underpinning the exceptional genetic and phenotypic plasticity associated with host adaptation in a pandemic clonal lineage of a eukaryotic plant pathogen. We observed that the asexual P. infestans lineage EC-1 can exhibit phenotypic plasticity in the absence of apparent genetic mutations resulting in virulence on a potato carrying the Rpi-vnt1.1 gene. Such variant alleles may be epialleles that arose through epigenetic changes in the underlying genes.


Coordinated multitissue transcriptional and plasma metabonomic profiles following acute caloric restriction in mice.
Caloric restriction (CR) increases healthy life span in a range of organisms. The underlying mechanisms are not understood but appear to include changes in gene expression, protein function, and metabolism. Recent studies demonstrate that acute CR alters mortality rates within days in flies. Multitissue transcriptional changes and concomitant metabolic responses to acute CR have not been described. We generated whole genome RNA transcript profiles in liver, skeletal muscle, colon, and hypothalamus and simultaneously measured plasma metabolites using proton nuclear magnetic resonance in mice subjected to acute CR. Liver and muscle showed increased gene expressions associated with fatty acid metabolism and a reduction in those involved in hepatic lipid biosynthesis. Glucogenic amino acids increased in plasma, and gene expression for hepatic gluconeogenesis was enhanced. Increased expression of genes for hormone-mediated signaling and decreased expression of genes involved in protein binding and development occurred in hypothalamus. Cell proliferation genes were decreased and cellular transport genes increased in colon. Acute CR captured many, but not all, hepatic transcriptional changes of long-term CR. Our findings demonstrate a clear transcriptional response across multiple tissues during acute CR, with congruent plasma metabolite changes. Liver and muscle switched gene expression away from energetically expensive biosynthetic processes toward energy conservation and utilization processes, including fatty acid metabolism and gluconeogenesis. Both muscle and colon switched gene expression away from cellular proliferation. Mice undergoing acute CR rapidly adopt many transcriptional and metabolic changes of long-term CR, suggesting that the beneficial effects of CR may require only a short-term reduction in caloric intake.


ATR pathway inhibition is synthetically lethal in cancer cells with ERCC1 deficiency.
The DNA damage response kinase ATR and its effector kinase CHEK1 are required for cancer cells to survive oncogene-induced replication stress. ATR inhibitors exhibit synthetic lethal interactions, with deficiencies in the DNA damage response enzymes ATM and XRCC1 and with overexpression of the cell cycle kinase cyclin E. Here, we report a systematic screen to identify synthetic lethal interactions with ATR pathway-targeted drugs, rationalized by their predicted therapeutic utility in the oncology clinic. We found that reduced function in the ATR pathway itself provided the strongest synthetic lethal interaction. In addition, we found that loss of the structure-specific endonuclease ERCC1-XPF (ERCC4) is synthetic lethal with ATR pathway inhibitors. ERCC1-deficient cells exhibited elevated levels of DNA damage, which was increased further by ATR inhibition. When treated with ATR or CHEK1 inhibitors, ERCC1-deficient cells were arrested in S-phase and failed to complete cell-cycle transit even after drug removal. Notably, triple-negative breast cancer cells and non-small cell lung cancer cells depleted of ERCC1 exhibited increased sensitivity to ATR pathway-targeted drugs. Overall, we concluded that ATR pathway-targeted drugs may offer particular utility in cancers with reduced ATR pathway function or reduced levels of ERCC4 activity.


Measurement variability in duplex scan assessment of carotid atherosclerosis.
The reproducibility of duplex scan measures of carotid atherosclerosis was evaluated as part of a study assessing the prevalence of carotid disease in elderly adults. Doppler measures of blood flow velocity were used to evaluate disease severity, and extent of carotid plaque was scored from the B-mode image. A reader assigned a grade from 0 to 3 to each of seven segments in the carotid system, based on the number and size of lesions present. Reproducibility data were obtained from 30 study participants who underwent a repeat scan by a second sonographer. Each scan was then scored by two readers. Doppler measures of blood flow velocity were found to be highly reproducible, with intraclass correlation coefficients of 0.81 for the common carotid artery, 0.84 for the internal carotid artery, and 0.77 for the internal carotid artery velocity to common carotid artery velocity ratio. Reproducibility of plaque grade was evaluated using segment as the unit of analysis, and both sonographer and reader variation were analyzed. When readers differed perfect agreement was achieved in 84% of the segments (K = 0.67), and when sonographers differed perfect agreement was obtained in 78% of the segments (K = 0.56). When both sonographer and reader differed, perfect agreement was obtained in 77% of the segments (K = 0.53). The plaque index, created by summing plaque grades from selected segments, was highly reproducible, with an intraclass correlation coefficient of 0.86. The duplex scan protocol described here provides reliable measures of both extent and severity of carotid disease that are appropriate for use in cross-sectional studies.


Black bean anthocyanin-rich extracts as food colorants: Physicochemical stability and antidiabetes potential.
Black beans contain anthocyanins that could be used as colorants in foods with associated health benefits. The objective was to optimize anthocyanins extraction from black bean coats and evaluate their physicochemical stability and antidiabetes potential. Optimal extraction conditions were 24% ethanol, 1:40 solid-to-liquid ratio and 29°C (P<0.0001). Three anthocyanins were identified by MS ions, delphinidin-3-O-glucoside (465.1m/z), petunidin-3-O-glucoside (479.1m/z) and malvidin-3-O-glucoside (493.1m/z). A total of 32mg of anthocyanins were quantified per gram of dry extract. Bean anthocyanins were stable at pH 2.5 and low-temperature 4°C (89.6%), with an extrapolated half-life of 277days. Anthocyanin-rich extracts inhibited α-glucosidase (37.8%), α-amylase (35.6%), dipeptidyl peptidase-IV (34.4%), reactive oxygen species (81.6%), and decreased glucose uptake. Black bean coats are a good source of anthocyanins and other phenolics with the potential to be used as natural-source food colorants with exceptional antidiabetes potential.


Stabilin-1 and Stabilin-2 are specific receptors for the cellular internalization of phosphorothioate-modified antisense oligonucleotides (ASOs) in the liver.
Phosphorothioate (PS)-modified antisense oligonucleotides (ASOs) have been extensively investigated over the past three decades as pharmacological and therapeutic agents. One second generation ASO, Kynamro™, was recently approved by the FDA for the treatment of homozygous familial hypercholesterolemia and over 35 second generation PS ASOs are at various stages of clinical development. In this report, we show that the Stabilin class of scavenger receptors, which were not previously thought to bind DNA, do bind and internalize PS ASOs. With the use of primary cells from mouse and rat livers and recombinant cell lines each expressing Stabilin-1 and each isoform of Stabilin-2 (315-HARE and 190-HARE), we have determined that PS ASOs bind with high affinity and these receptors are responsible for bulk, clathrin-mediated endocytosis within the cell. Binding is primarily dependent on salt-bridge formation and correct folding of the intact protein receptor. Increased internalization rates also enhanced ASO potency for reducing expression of the non-coding RNA Malat-1, in Stabilin-expressing cell lines. A more thorough understanding of mechanisms by which ASOs are internalized in cells and their intracellular trafficking pathways will aid in the design of next generation antisense agents with improved therapeutic properties.


Pilot study investigating infant vagal reactivity and visual behavior during object perception.
The authors examined whether changes in vagal tone were related to infant visual attention during auditory and visual events paired (synchronous) and not paired (asynchronous) in time. They predicted that infants would demonstrate greater visual attention to the synchronous slideshow and that vagal tone would decrease with visual attention. Nineteen infants, 3.5 months old, watched computer-generated synchronous or asynchronous slideshows of auditory and visual stimuli. Visual behavior and vagal tone data were collected. Vagal tone reflects physiological responses during attention or exposure to mild stressors. Repeated-measures analysis of variance examined differences in vagal tone across conditions. Visual behavior did not differ between the synchronous and asynchronous slideshow conditions. Vagal tone was significantly lower during the asynchronous slideshow. Infants may discriminate synchronous from asynchronous stimuli without changing visual behavior. Implications related to play with toys or objects are discussed.


Material Flow Analysis with Multiple Material Characteristics to Assess the Potential for Flat Steel Prompt Scrap Prevention and Diversion without Remelting.
Thirty-two percent of the liquid metal used to make flat steel products in Europe does not end up in a final product. Sixty percent of this material is instead scrapped during manufacturing and the remainder during fabrication of finished steel products. Although this scrap is collected and recycled, remelting this scrap requires approximately 2 MWh/t, but some of this material could instead be diverted for use in other applications without remelting. However, this diversion depends not just on the mass of scrapped steel but also on its material characteristics. To enhance our understanding of the potential for such scrap diversion, this paper presents a novel material flow analysis of flat steel produced in Europe in 2013. This analysis considers the flow of steel characterized not only by mass but, for the first time, also by grade, thickness, and coating. The results show that thin-gauge galvanized drawing steel is the most commonly demanded steel grade across the industry, and most scrap of this grade is generated by the automotive industry. There are thus potential opportunities for preventing and diverting scrap of this grade. We discuss the role of the geometric compatibility of parts and propose tessellating blanks for various car manufacturers in the same coil of steel to increase the utilization rates of steel.


Adverse effects of prolonged hyperventilation in patients with severe head injury: a randomized clinical trial.
There is still controversy over whether or not patients should be hyperventilated after traumatic brain injury, and a randomized trial has never been conducted. The theoretical advantages of hyperventilation are cerebral vasoconstriction for intracranial pressure (ICP) control and reversal of brain and cerebrospinal fluid (CSF) acidosis. Possible disadvantages include cerebral vasoconstriction to such an extent that cerebral ischemia ensues, and only a short-lived effect on CSF pH with a loss of HCO3-buffer from CSF. The latter disadvantage might be overcome by the addition of the buffer tromethamine (THAM), which has shown some promise in experimental and clinical use. Accordingly, a trial was performed with patients randomly assigned to receive normal ventilation (PaCO2 35 +/- 2 mm Hg (mean +/- standard deviation): control group), hyperventilation (PaCO2 25 +/- 2 mm Hg: HV group), or hyperventilation plus THAM (PaCO2 25 +/- 2 mm Hg: HV + THAM group). Stratification into subgroups of patients with motor scores of 1-3 and 4-5 took place. Outcome was assessed according to the Glasgow Outcome Scale at 3, 6, and 12 months. There were 41 patients in the control group, 36 in the HV group, and 36 in the HV + THAM group. The mean Glasgow Coma Scale score for each group was 5.7 +/- 1.7, 5.6 +/- 1.7, and 5.9 +/- 1.7, respectively; this score and other indicators of severity of injury were not significantly different. A 100% follow-up review was obtained. At 3 and 6 months after injury the number of patients with a favorable outcome (good or moderately disabled) was significantly (p less than 0.05) lower in the hyperventilated patients than in the control and HV + THAM groups. This occurred only in patients with a motor score of 4-5. At 12 months posttrauma this difference was not significant (p = 0.13). Biochemical data indicated that hyperventilation could not sustain alkalinization in the CSF, although THAM could. Accordingly, cerebral blood flow (CBF) was lower in the HV + THAM group than in the control and HV groups, but neither CBF nor arteriovenous difference of oxygen data indicated the occurrence of cerebral ischemia in any of the three groups. Although mean ICP could be kept well below 25 mm Hg in all three groups, the course of ICP was most stable in the HV + THAM group. It is concluded that prophylactic hyperventilation is deleterious in head-injured patients with motor scores of 4-5.(ABSTRACT TRUNCATED AT 400 WORDS)


The OpenMRS Implementers Network.
OpenMRS (www.openmrs.org) is a configurable open source electronic medical record application developed and maintained by a large network of open source developers coordinated by the Regenstrief Institute and Partners in Health and mainly used for HIV patient and treatment information management in Africa. Our objective is to develop an open Implementers Network for OpenMRS to provide regional support for the growing number of OpenMRS implementations in Africa and to include African developers and implementers in the future growth of OpenMRS. We have developed the OpenMRS Implementers Network using a dedicated Wiki site and e-mail server. We have also organized annual meetings in South Africa and regional training courses at African locations where OpenMRS is being implemented. An OpenMRS Internship program has been initiated and we have started collaborating with similar networks and projects working in Africa. To evaluate its potential, OpenMRS was implemented initially at one site in South Africa by a single implementer using a downloadable OpenMRS application and only the OpenMRS Implementers Network for support. The OpenMRS Implementers Network Wiki and list server have grown into effective means of providing implementation support and forums for exchange of implementation experiences. The annual OpenMRS Implementers meeting has been held in South Africa for the past three years and is attracting successively larger numbers of participants with almost 200 implementers and developers attending the 2008 meeting in Durban, South Africa. Six African developers are presently registered on the first intake of the OpenMRS Internship program. Successful collaborations have been started with several African developer groups and projects initiated to develop interoperability between OpenMRS and various applications. The South African OpenMRS Implementer group successfully configured, installed and maintained an integrated HIV/TB OpenMRS application without significant programming support. Since then, this model has been replicated in several other African sites. The OpenMRS Implementers Network has contributed substantially to the growth and sustainability of OpenMRS in Africa and has become a useful way of including Africans in the development and implementation of OpenMRS in developing countries. The Network provides valuable support and enables a basic OpenMRS application to be implemented in the absence of onsite programmers.


A pilot study on in vitro solubility of phosphorus from mineral sources, feed ingredients and compound feed for pigs, poultry, dogs and cats.
Excess phosphorus (P) as seen in cat foods can have a negative effect on health (Dobenecker, Webel, Reese, & Kienzle, ; Pastoor, Klooster, Mathot, & Beynen, ). P surpluses may affect the environment, and economics in food producing animals, whereas marginal supply may impair performance and health. P can only be absorbed if it is soluble. Solubility of feed P in water and weak acid solution-as a precondition for absorption-was investigated in feed for dogs, cats, pigs and poultry. Different P containing mineral compounds (Ca(H2 PO4 )2 , CaHPO4 •2H2 O, Ca4 Na(PO4 )3 , KH2 PO4 , K4 P2 O7 , NaH2 PO4 , Na5 P3 O10 (29 samples), as well as eight different ingredients such as wheat or meat, 64 compound feeds for pig and poultry, eight complete dry and 13 complete moist dog foods, 25 complete moist cat foods and 29 experimental diets were analysed for P solubility. Finely ground feeds were soaked in water or hydrochloric acid (0.4%) for 1 and 90 min. The samples were centrifuged and the supernatant was analysed for P (photometric vanadate molybdate method after wet ashing). The solubility of P from inorganic sources reflected the solubility of the main compound of the feed grade material. "organic" ingredients, such as fish meal or meat, showed a lower P solubility than inorganic sources. Most ingredients from animal origin (exception fish meal) had a higher P solubility than those from plant origin. When inorganic and "organic" P sources were mixed, the P solubility of the mixture reflected the P solubility and percentages of its compounds. In chicken, turkey and pig compound feed the percentage of acid soluble P increased with increasing P content. Pet moist food showed high percentages of water-soluble P. The results show that the method is suitable to obtain data on water and acid solubility of P in feed and ingredients.


Wastewater garden--a system to treat wastewater with environmental benefits to community.
Many communities and villages around the world face serious problems with lack of sanitation especially in disposing of the wastewater-black water and grey water from the houses, or wash outs from animal rearing sheds. Across the world diverting wastewater to the surroundings or to the public spaces are not uncommon. This is responsible for contaminating drinking water sources causing health risks and environmental degradation as they become the breeding grounds of mosquitoes and pathogens. Lack of collection and treatment facilities or broken down sewage systems noticed throughout the developing world are associated with this situation. Diverting the wastewater to trees and vegetable gardens was historically a common practice. However the modern world has an array of problems associated with such disposal such as generation of large quantity of wastewater, unavailability of space for onsite disposal or treatment and increase in population. This paper considers the wastewater garden as a means for wastewater treatment and to improve the vegetation and biodiversity of rural areas. This can also be implemented in urban areas in association with parks and open spaces. This also highlights environmental safety in relation to the nutrient, pathogen and heavy metal content of the wastewater. The possibilities of different types of integration and technology that can be adopted for wastewater gardens are also discussed.


A taxonomy of medical uncertainties in clinical genome sequencing.
Clinical next-generation sequencing (CNGS) is introducing new opportunities and challenges into the practice of medicine. Simultaneously, these technologies are generating uncertainties of an unprecedented scale that laboratories, clinicians, and patients are required to address and manage. We describe in this report the conceptual design of a new taxonomy of uncertainties around the use of CNGS in health care. Interviews to delineate the dimensions of uncertainty in CNGS were conducted with genomics experts and themes were extracted in order to expand on a previously published three-dimensional taxonomy of medical uncertainty. In parallel, we developed an interactive website to disseminate the CNGS taxonomy to researchers and engage them in its continued refinement. The proposed taxonomy divides uncertainty along three axes-source, issue, and locus-and further discriminates the uncertainties into five layers with multiple domains. Using a hypothetical clinical example, we illustrate how the taxonomy can be applied to findings from CNGS and used to guide stakeholders through interpretation and implementation of variant results. The utility of the proposed taxonomy lies in promoting consistency in describing dimensions of uncertainty in publications and presentations, to facilitate research design and management of the uncertainties inherent in the implementation of CNGS.Genet Med advance online publication 19 January 2017.


Differential priming of RNA templates during cDNA synthesis markedly affects both accuracy and reproducibility of quantitative competitive reverse-transcriptase PCR.
Quantitative competitive reverse-transcriptase PCR is the most sensitive method for studying gene expression. To investigate whether the accuracy of the calculated target mRNA copy number is affected by the cDNA priming process, we utilized primers of different lengths, concentrations and primer sequences to prime cDNA synthesis reactions. Our results show a approximately 19-fold increase in the calculated mRNA copy number from cDNA synthesis reactions primed with random hexamers (P<0.001, n=4), and a approximately 4-fold increase in copy number with a specific hexamer (P<0.001, n=4) compared with that obtained with a 22-mer-sequence-specific primer. The increase in calculated mRNA copy number obtained by priming cDNA synthesis with the shorter specific and non-specific primers could be explained largely by the synthesis of truncated standard cDNA molecules lacking a requisite binding site for amplification with PCR primers. Since these truncated standard cDNA molecules could not be amplified and standard RNA is used to quantify target mRNA copy number, this phenomenon resulted in overestimation of target mRNA copy number. In conclusion, accurate determination of target mRNA copy number is most likely if a long specific antisense primer is used to prime cDNA synthesis.


Suffocation using plastic bags: a retrospective study of suicides in Ontario, Canada.
One hundred and ten cases of suicidal suffocation using a plastic bag were identified in the files of the Office of the Chief Coroner of Ontario, Canada, between 1993 and 1997. The records were reviewed to determine the demographic characteristics of this group compared with all cases of suicide in Ontario, the scene information, autopsy findings and toxicology results. Most suicides occurred in people over 60 years of age, with older women making up a considerable proportion of cases as compared with other methods of suicide. In 40% of cases the deceased was suffering from a serious illness. Autopsy findings were usually minimal, with facial, conjunctival and visceral petechiae present in a minority of cases. One or more drugs were detected in the blood in 92.6% of cases where toxicologic testing was performed. Benzodiazepines, diphenhydramine and antidepressants were the most common drugs found, with diphenhydramine the most common drug present at an elevated concentration. Information at the scene from "right to die" societies was uncommon. One quarter of decedents took additional measures, besides the use of drugs or alcohol, to ensure the rapidity, certainty or comfort of their death. This study further elucidates the characteristics of this uncommon method of suicide. It emphasizes additional scene findings, such as the presence of dust masks, physical restraints and modification of the plastic bag that may be of use to death investigators in determining the correct manner of death.


Introduction: new tools for enhancing collaborative endometriosis research.
This issue of Fertility and Sterility contains four articles by the World Endometriosis Research Foundation whose present objective is global standardization of the collection of phenotypic data and biological samples, designated as the Endometriosis Phenome and Biobanking Harmonisation Project. The aim is to facilitate large-scale international, multicenter trials that are robust, and will result in biomarker and treatment targets to advance research in endometriosis.


AT-rich repeats associated with chromosome 22q11.2 rearrangement disorders shape human genome architecture on Yq12.
Low copy repeats (LCRs; segmental duplications) constitute approximately 5% of the sequenced human genome. Nonallelic homologous recombination events between LCRs during meiosis can lead to chromosomal rearrangements responsible for many genomic disorders. The 22q11.2 region is susceptible to recurrent and nonrecurrent deletions, duplications as well as translocations that are mediated by LCRs termed LCR22s. One particular DNA structural element, a palindromic AT-rich repeat (PATRR) present within LCR22-3a, is responsible for translocations. Similar AT-rich repeats are present within the two largest LCR22s, LCR22-2 and LCR22-4. We provide direct sequence evidence that the AT-rich repeats have altered LCR22 organization during primate evolution. The AT-rich repeats are surrounded by a subtype of human satellite I (HSAT I), and an AluSc element, forming a 2.4-kb tripartite structure. Besides 22q11.2, FISH and PCR mapping localized the tripartite repeat within heterochromatic, unsequenced regions of the genome, including the pericentromeric regions of the acrocentric chromosomes and the heterochromatic portion of Yq12 in humans. The repeat is also present on autosomes but not on chromosome Y in other hominoid species, suggesting that it has duplicated on Yq12 after speciation of humans from its common ancestor. This demonstrates that AT-rich repeats have shaped or altered the structure of the genome during evolution.


Clinical and pH-metric characteristics of gastro-oesophageal reflux secondary to cows' milk protein allergy.
The primary aim was to assess whether there were differences in symptoms, laboratory data, and oesophageal pH-metry between infants with primary gastro-oesophageal reflux and those with reflux secondary to cows' milk protein allergy (CMPA). 96 infants (mean(SD) age 7.8(2.0) months) with either primary gastro-oesophageal reflux, reflux with CMPA, CMPA only, or none of these (controls) were studied. Symptoms, immunochemical data, and oesophageal pH were compared between the four groups and the effect of a cows' milk protein-free diet on the severity of symptoms was also assessed. 14 out of 47(30%) infants with gastro-oesophageal reflux had CMPA. These infants had similar symptoms to those with primary gastro-oesophageal reflux but higher concentrations of total IgE and circulating eosinophils (p < 0.005) and IgG anti-beta lactoglobulin (p < 0.003). A progressive constant reduction in oesophageal pH at the end of a feed, which continued up to the next feed, was seen in 12 out of 14 patients with gastro-oesophageal reflux secondary to CMPA and in 24 of 25 infants with CMPA only. No infants with primary gastro-oesophageal reflux and none of the controls had this pattern. A cows' milk protein-free diet was associated with a significant improvement in symptoms only in infants with gastro-oesophageal reflux with CMPA. A characteristic oesophageal pH pattern is useful in distinguishing infants with gastro-oesophageal reflux associated with CMPA.


Gene expression in epithelial cells in response to pneumovirus infection.
Respiratory syncytial virus (RSV) and pneumonia virus of mice (PVM) are viruses of the family Paramyxoviridae, subfamily pneumovirus, which cause clinically important respiratory infections in humans and rodents, respectively. The respiratory epithelial target cells respond to viral infection with specific alterations in gene expression, including production of chemoattractant cytokines, adhesion molecules, elements that are related to the apoptosis response, and others that remain incompletely understood. Here we review our current understanding of these mucosal responses and discuss several genomic approaches, including differential display reverse transcription-polymerase chain reaction (PCR) and gene array strategies, that will permit us to unravel the nature of these responses in a more complete and systematic manner.


Analysis of 3D genomic interactions identifies candidate host genes that transposable elements potentially regulate.
The organization of chromatin in the nucleus plays an essential role in gene regulation. About half of the mammalian genome comprises transposable elements. Given their repetitive nature, reads associated with these elements are generally discarded or randomly distributed among elements of the same type in genome-wide analyses. Thus, it is challenging to identify the activities and properties of individual transposons. As a result, we only have a partial understanding of how transposons contribute to chromatin folding and how they impact gene regulation. Using PCR and Capture-based chromosome conformation capture (3C) approaches, collectively called 4Tran, we take advantage of the repetitive nature of transposons to capture interactions from multiple copies of endogenous retrovirus (ERVs) in the human and mouse genomes. With 4Tran-PCR, reads are selectively mapped to unique regions in the genome. This enables the identification of transposable element interaction profiles for individual ERV families and integration events specific to particular genomes. With this approach, we demonstrate that transposons engage in long-range intra-chromosomal interactions guided by the separation of chromosomes into A and B compartments as well as topologically associated domains (TADs). In contrast to 4Tran-PCR, Capture-4Tran can uniquely identify both ends of an interaction that involve retroviral repeat sequences, providing a powerful tool for uncovering the individual transposable element insertions that interact with and potentially regulate target genes. 4Tran provides new insight into the manner in which transposons contribute to chromosome architecture and identifies target genes that transposable elements can potentially control.


Sensorineural hearing loss after ossicular manipulation and drill-generated acoustic trauma in type I tympanoplasty with and without mastoidectomy: A series of 51 cases.
Recognized causes of high-frequency sensorineural hearing loss (SNHL) after tympanoplasty with and without mastoidectomy include acoustic trauma from manipulation of the ossicles, the noise generated by suctioning and, in the case of mastoidectomy, the noise generated by temporal bone drilling. We conducted a retrospective study to identify the effects of ossicular manipulation and mastoid drilling on high-frequency SNHL. Our study population was made up of 51 patients-20 males and 31 females, aged 10 to 59 years (mean: 28.5). Of this group, 26 patients had undergone a unilateral over-under tympanoplasty only (tympanoplasty group) and 25 had undergone a unilateral tympanoplasty plus additional mastoid surgery (mastoidectomy group). Bone-conduction audiograms were obtained pre- and postoperatively; the latter were obtained within 24 hours after surgery and again at 6 months of follow-up. In the tympanoplasty group, a significant SNHL, primarily at 2 kHz, was seen in 6 patients (23%) at 24 hours, but at 6 months there was no depression of bone-conduction thresholds. In the mastoidectomy group, a significant SNHL, primarily at 2 and 4 kHz, occurred in 12 patients (48%) at 24 hours, and bone-conduction deterioration was still present in 4 patients (16%) 6 months after surgery. The difference between the preoperative audiograms and the 6-month audiograms in both groups was statistically significant (p = 0.034). We conclude that (1) over-under tympanoplasty, which requires significant manipulation of the ossicles, can cause temporary SNHL after surgery, and (2) prolonged exposure to the noise generated by mastoid drilling can result in permanent SNHL.


Fluorescent and colorimetric ion probes based on conjugated oligopyrroles.
Metal ions and anions play important roles in many industrial and biochemical processes, and thus it is highly desired to detect them in the relevant systems. Small organic molecule based sensors for selective and sensitive detection of target ions show the advantages of low cost, high sensitivity and convenient implementation. In this area, pyrrole has incomparable advantages. It can be easily incorporated into linear and macrocyclic conjugated structures such as dipyrrins, porphyrins, and N-confused porphyrins, which may utilize the imino N and amino NH moieties for binding metal ions and anions, respectively. In this tutorial review, we focus on representative examples to describe the design, syntheses, sensing mechanisms, and applications of the conjugated oligopyrroles. These compounds could be used as colorimetric or fluorescent ion probes, with the advantages of vivid colour and fluorescence changes, easy structural modification and functionalization, and tunable emission wavelengths. Compared with normal porphyrins, simple di- and tripyrrins, as well as some porphyrinoids are more suitable for designing fluorescence "turn-on" metal probes, because they may exhibit flexible confirmations, and metal coordination will improve the rigidity, resulting in vivid fluorescence enhancement. It is noteworthy that the oligopyrrolic moieties may simultaneously act as the binding unit as well as the reporting moiety, which simplifies the design and syntheses of the probes.


The clinicopathological characteristics and survival outcomes of primary expansile vs. infiltrative mucinous ovarian adenocarcinoma: a retrospective study sharing the experience of a tertiary centre.
Mucinous ovarian carcinomas (MOCs) are rare ovarian tumours accounting for 3% of all epithelial ovarian carcinomas (EOCs). They are either expansile or infiltrative, based on the tumour's histological pattern of invasion. MOCs have a distinct molecular profile, natural history, chemo-sensitivity, and prognosis compared to other EOCs. The aim of this study was to describe patient and tumour characteristics, as well as survival outcomes of expansile and infiltrative primary MOCs. This was a retrospective cohort study conducted at a tertiary cancer centre. Patients had surgery for primary MOC between Jul 1, 2010 and Oct 28, 2022. All patients discussed at the Oxford multidisciplinary team (MDT) meeting with a diagnosis of MOC were included. We excluded patients with mucinous metastatic carcinoma (MMC), dual histological diagnoses, those who died before treatment was initiated, and patients with incomplete records. A total of 47 patients were identified and 14 were excluded. Out of the remaining 33 MOCs, 23 (70.6%) were expansile and 10 (30.4%) were infiltrative. The median follow-up was 37 months (95% CI: 14.1-69.8). Patients with infiltrative tumours were older than those with expansile tumours (median age 62 vs. 55 years, P=0.049). Infiltrative tumours were diagnosed at a more advanced International Federation of Gynaecology and Obstetrics (FIGO) stage compared to expansile tumours: FIGO stage II/III 50% vs. 8.2% (P=0.002). We found paired-box gene 8 (PAX8) more frequently expressed in expansile tumours (75% vs. 37.5%, P=0.099). Adjuvant treatment was administered in 50% of patients with infiltrative disease, compared to only 13% of those with expansile disease (P=0.036). 80% of patients who have relapsed had received adjuvant chemotherapy, compared to 17.2% of patients without relapse (P=0.012). At 3 years, there was a statistically significant difference in progression-free survival (PFS) (94.7% vs. 65.6%, P=0.02) between the expansile and infiltrative groups, but no difference in overall survival (OS) (88.8% vs. 90%, P=0.875). Patients with infiltrative tumours were older, more likely to have bilateral tumours and more likely to have an advanced FIGO stage at diagnosis. Adjuvant treatment was more likely to be administered to patients with infiltrative tumours, however, this did not prevent relapse. PFS at 3 years was significantly higher in patients with expansile tumours. PAX8 was more frequently expressed by expansile tumours.


Murine lung tumor response after exposure to cigarette mainstream smoke or its particulate and gas/vapor phase fractions.
Knowledge on mechanisms of smoking-induced tumorigenesis and on active smoke constituents may improve the development and evaluation of chemopreventive and therapeutic interventions, early diagnostic markers, and new and potentially reduced-risk tobacco products. A suitable laboratory animal disease model of mainstream cigarette smoke inhalation is needed for this purpose. In order to develop such a model, A/J and Swiss SWR/J mouse strains, with a genetic susceptibility to developing lung adenocarcinoma, were whole-body exposed to diluted cigarette mainstream smoke at 0, 120, and 240 mg total particulate matter per m(3) for 6h per day, 5 days per week. Mainstream smoke is the smoke actively inhaled by the smoker. For etiological reasons, parallel exposures to whole smoke fractions (enriched for particulate or gas/vapor phase) were performed at the higher concentration level. After 5 months of smoke inhalation and an additional 4-month post-inhalation period, both mouse strains responded similarly: no increase in lung tumor multiplicity was seen at the end of the inhalation period; however, there was a concentration-dependent tumorigenic response at the end of the post-inhalation period (up to 2-fold beyond control) in mice exposed to the whole smoke or the particulate phase. Tumors were characterized mainly as pulmonary adenomas. At the end of the inhalation period, epithelial hyperplasia, atrophy, and metaplasia were found in the nasal passages and larynx, and cellular and molecular markers of inflammation were found in the bronchoalveolar lavage fluid. These inflammatory effects were mostly resolved by the end of the post-inhalation period. In summary, these mouse strains responded to mainstream smoke inhalation with enhanced pulmonary adenoma formation. The major tumorigenic potency resided in the particulate phase, which is contrary to the findings published for environmental tobacco smoke surrogate inhalation in these mouse models.


Nucleolar diameter and microvascular factors as independent predictors of mortality from malignant melanoma of the choroid and ciliary body.
To determine whether nucleolar diameter and microvascular factors are independent predictors of mortality in malignant uveal melanoma of the choroid and ciliary body. A population-based retrospective cohort study was conducted of melanoma-specific and all-cause mortality in 167 consecutive patients who had an eye enucleated because of choroidal and ciliary body melanoma from 1972 through 1981. The largest nucleoli were measured from digital photographs of silver-stained tumors along a central 5-mm-wide linear field parallel to the base of the tumor. The mean of the 10 largest nucleoli (MLN) was calculated. Microvascular loops and networks and microvascular density (MVD) were assessed. Kaplan-Meier and Cox regression analyses were performed. Associations between MLN and other variables were determined. The MLN could be determined in 126 (75%) melanomas. It ranged from 2.60 to 6.18 microm (median, 4.05). The association of large MLN with the presence of epithelioid cells (P = 0.017) and high MVD (P = 0.0053) was statistically significant. MLN was not significantly associated with tumor diameter and microvascular loops and networks. The 10-year melanoma-specific survival decreased with MLN (0.74, 0.60, and 0.42, arranged in tertiles; P = 0.0060), presence of loops and networks (P = 0.0001), and increasing MVD (P = 0.0001). By Cox regression, MLN was an independent predictor of survival, when adjusting in turn for presence of epithelioid cells, loops and networks, and MVD. In multivariate models with MVD, the independent prognostic information carried by MLN decreased, but the model as a whole was a better predictor of survival. The magnitude of this effect depended on whether MLN was modeled as a continuous or categorical variable. In this population-based data set, MLN and microvascular loops and networks were unrelated, independent predictors of survival. MLN and MVD were found to be partially interrelated. Multivariate models that included MVD in addition to MLN fitted better with observed melanoma-specific survival than models that excluded MVD.


Gender differences in the processes of change for condom use: patterns across stages of change in crack cocaine users.
Interventions to increase condom use in crack users have had mixed results. For interventions to achieve greater success, the mechanisms of behavior change in this population need to be understood. One mechanism, the processes of change, was examined across stages of change for condom use. Results from the analysis of variance for males and females revealed that stage of change was associated with different levels of three experiential processes: consciousness raising, social liberation and self-reevaluation. However, these analyses found that male and females seem to have different patterns of behavioral process use. Specifically, females in the preparation stage were different from those in precontemplation, whereas this difference was not pronounced in males. In general, people had high levels of experimental processes in every stage of change. The patterns of behavior process use mimicked patterns found for other behaviors with a linear increase across the stages of change. This may indicate that for maintaining condom use, more emotional and behavioral activities are required throughout the process of acquisition and maintenance than are necessary for other health-related activities. Implications of this research are that interventions for increasing condom use in drug users may target behavioral steps differently for males and females.


Epidemiological characteristics, safety and efficacy of medical cannabis in the elderly.
There is a substantial growth in the use of medical cannabis in recent years and with the aging of the population, medical cannabis is increasingly used by the elderly. We aimed to assess the characteristics of elderly people using medical cannabis and to evaluate the safety and efficacy of the treatment. A prospective study that included all patients above 65 years of age who received medical cannabis from January 2015 to October 2017 in a specialized medical cannabis clinic and were willing to answer the initial questionnaire. Outcomes were pain intensity, quality of life and adverse events at six months. During the study period, 2736 patients above 65 years of age began cannabis treatment and answered the initial questionnaire. The mean age was 74.5 ± 7.5 years. The most common indications for cannabis treatment were pain (66.6%) and cancer (60.8%). After six months of treatment, 93.7% of the respondents reported improvement in their condition and the reported pain level was reduced from a median of 8 on a scale of 0-10 to a median of 4. Most common adverse events were: dizziness (9.7%) and dry mouth (7.1%). After six months, 18.1% stopped using opioid analgesics or reduced their dose. Our study finds that the therapeutic use of cannabis is safe and efficacious in the elderly population. Cannabis use may decrease the use of other prescription medicines, including opioids. Gathering more evidence-based data, including data from double-blind randomized-controlled trials, in this special population is imperative.


DNA viruses: the really big ones (giruses).
Viruses with genomes greater than 300 kb and up to 1200 kb are being discovered with increasing frequency. These large viruses (often called giruses) can encode up to 900 proteins and also many tRNAs. Consequently, these viruses have more protein-encoding genes than many bacteria, and the concept of small particle/small genome that once defined viruses is no longer valid. Giruses infect bacteria and animals although most of the recently discovered ones infect protists. Thus, genome gigantism is not restricted to a specific host or phylogenetic clade. To date, most of the giruses are associated with aqueous environments. Many of these large viruses (phycodnaviruses and Mimiviruses) probably have a common evolutionary ancestor with the poxviruses, iridoviruses, asfarviruses, ascoviruses, and a recently discovered Marseillevirus. One issue that is perhaps not appreciated by the microbiology community is that large viruses, even ones classified in the same family, can differ significantly in morphology, lifestyle, and genome structure. This review focuses on some of these differences than on extensive details about individual viruses.


A systematic review of the unmet supportive care needs of men on active surveillance for prostate cancer.
Understanding the unmet supportive care needs of men on active surveillance for prostate cancer may enable researchers and health professionals to better support men and prevent discontinuation when there is no evidence of disease progression. This review aimed to identify the specific unmet supportive care needs of men on active surveillance. A systematic review following PRISMA guidelines was conducted. Databases (Pubmed, Embase, PsycINFO, and CINAHL) were searched to identify qualitative and/or quantitative studies that reported unmet needs specific to men on active surveillance. Quality appraisals were conducted before results were narratively synthesised. Of the 3613 unique records identified, only eight articles were eligible (five qualitative and three cross-sectional studies). Unmet Informational, Emotional/Psychological, Social, and "Other" needs were identified. Only three studies had a primary aim of investigating unmet supportive care needs. Small active surveillance samples, use of nonvalidated measures, and minimal reporting of author reflexivity in qualitative studies were the main quality issues identified. The unmet needs of men on active surveillance is an underresearched area. Preliminary evidence suggests the information available and provided to men during active surveillance is perceived as inadequate and inconsistent. Men may also be experiencing unmet psychological/emotional, social, and other needs; however, further representative, high-quality research is required to understand the magnitude of this issue. Reporting results specific to treatment type and utilising relevant theories/models (such as the social ecological model [SEM]) is recommended to ensure factors that may facilitate unmet needs are appropriately considered and reported.


Two asymmetric hadamard transform spectrometers.
We develop the theory of operation for dispersive spectrometers that modulate radiation at both the entrance and exit apertures by means of Hadamard codes. Specifically, we examine the operation of instruments illuminated by a beam of radiation known to be homogeneous. In this case, all spatial information obtained in the operation of the instrument can be effectively suppressed at no loss of spectral performance and at a considerable reduction in the number of measurements that need to be made. A particularly interesting instrument which mocks a monochromator is described. The spectrum is directly obtained from the data by simply subtracting a constant intensity value from all readings. This instrument bears a resemblance to the Girard grill spectrometer. We describe the construction and operation of an instrument that has been tested in both modes of operation and show some of the spectra obtained.


Estimation of Tree Size Diversity Using Object Oriented Texture Analysis and Aster Imagery.
This study investigates the potential of object-based texture parameters extracted from 15m spatial resolution ASTER imagery for estimating tree size diversity in a Mediterranean forested landscape in Turkey. Tree size diversity based on tree basal area was determined using the Shannon index and Gini Coefficient at the sampling plot level. Image texture parameters were calculated based on the grey level co-occurrence matrix (GLCM) for various image segmentation levels. Analyses of relationships between tree size diversity and texture parameters found that relationships between the Gini Coefficient and the GLCM values were the most statistically significant, with the highest correlation (r=0.69) being with GLCM Homogeneity values. In contrast, Shannon Index values were weakly correlated with image derived texture parameters. The results suggest that 15m resolution Aster imagery has considerable potential in estimating tree size diversity based on the Gini Coefficient for heterogeneous Mediterranean forests.


Pyrroloquinoline Quinone Resists Denervation-Induced Skeletal Muscle Atrophy by Activating PGC-1α and Integrating Mitochondrial Electron Transport Chain Complexes.
Denervation-mediated skeletal muscle atrophy results from the loss of electric stimulation and leads to protein degradation, which is critically regulated by the well-confirmed transcriptional co-activator peroxisome proliferator co-activator 1 alpha (PGC-1α). No adequate treatments of muscle wasting are available. Pyrroloquinoline quinone (PQQ), a naturally occurring antioxidant component with multiple functions including mitochondrial modulation, demonstrates the ability to protect against muscle dysfunction. However, it remains unclear whether PQQ enhances PGC-1α activation and resists skeletal muscle atrophy in mice subjected to a denervation operation. This work investigates the expression of PGC-1α and mitochondrial function in the skeletal muscle of denervated mice administered PQQ. The C57BL6/J mouse was subjected to a hindlimb sciatic axotomy. A PQQ-containing ALZET® osmotic pump (equivalent to 4.5 mg/day/kg b.w.) was implanted subcutaneously into the right lower abdomen of the mouse. In the time course study, the mouse was sacrificed and the gastrocnemius muscle was prepared for further myopathological staining, energy metabolism analysis, western blotting, and real-time quantitative PCR studies. We observed that PQQ administration abolished the denervation-induced decrease in muscle mass and reduced mitochondrial activities, as evidenced by the reduced fiber size and the decreased expression of cytochrome c oxidase and NADH-tetrazolium reductase. Bioenergetic analysis demonstrated that PQQ reprogrammed the denervation-induced increase in the mitochondrial oxygen consumption rate (OCR) and led to an increase in the extracellular acidification rate (ECAR), a measurement of the glycolytic metabolism. The protein levels of PGC-1α and the electron transport chain (ETC) complexes were also increased by treatment with PQQ. Furthermore, PQQ administration highly enhanced the expression of oxidative fibers and maintained the type II glycolytic fibers. This pre-clinical in vivo study suggests that PQQ may provide a potent therapeutic benefit for the treatment of denervation-induced atrophy by activating PGC-1α and maintaining the mitochondrial ETC complex in skeletal muscles.


Socioeconomic and dietary risk factors for anemia in children aged 6 to 59 months.
The objective of this study was to use statistical modeling to identify risk factors for anemia in children aged 6 to 59 months in the state of Pernambuco, covering socioeconomic and dietary aspects. The sample comprised 746 children aged between 6 and 59 months from the state of Pernambuco. Their hemoglobin was assayed and a 24-hour dietary recall performed. Risk of anemia was analyzed with relation to socioeconomic variables and to dietary intakes, using multivariate analysis models. The risk factors for anemia were: a high proportion of calories from cow's milk, low density of nonheme iron, low age and low maternal educational level; age was the most prominent factor, with children under 24 months exhibiting 3.61 times greater risk of being anemic than the older children. This study confirms the need for a clear picture of the dietary intake of children in Brazil, in order that associations with anemia can be better understood. Our results revealed that the dietary factors which were most responsible for risk of anemia were a greater proportion of calories from cow's milk and lower density of nonheme iron, in addition to age below 24 months and low maternal educational level.


High Bolus Tirofiban vs Abciximab in Acute STEMI Patients Undergoing Primary PCI - The Tamip Study.
Primary percutaneous coronary intervention (PCI) has been shown to be an effective therapy for patients with acute myocardial infarction (MI). Glycoprotein (GP) IIb/IIIa receptor blockers reduce thrombotic complications in patients undergoing PCI. Most available data relate to Reopro, which has been registered for this indication. GP IIb/IIIa reduce unfavorable outcome in U/A and non ST-elevation myocardial infarction (STEMI) patients. Only few studies focused on high dose Aggrastat for STEMI patients in the emergency department (ED) before PCI. The aim is to increase the patency during the time awaiting coronary angioplasty in patients with acute MI. To study the effect of upfront high bolus dose (HDR) of tirofiban on the extent of residual ST segment deviation 1 hour after primary PCI and the incidence of TIMI 3 flow of the infarct-related artery (IRA). A randomized, open label, single center study in the ED. A total of 90 patients with acute ST-elevation MI, diagnosed clinically by ECG criteria (ST segment elevation of >2 mm in two adjacent ECG leads), and with an expectation that a patient will undergo primary PCI. Patients were aged 21-85 years and all received heparin 5000 u, aspirin 160 mg, and Plavix 600 mg. Patients were divided in two groups (group I: triofiban high bolus vs group II: Reopro) with 45 patients in each group. In group I, high bolus triofiban 25 mcg/kg over 3 min was started in the ED with maintenance infusion of 0.15 mcg/ kg/min continued for 12 hours and transferred to cath lab for PCI. Patients in group II were transferred to cath lab, where a standard dose of Reopro was given with a bolus of 0.25 mcg/kg and maintenance infusion of 0.125 mcg/kg/min over 12 hours. ST segment resolution and TIMI flow were evaluated in both groups before and after PCI. Thirty-five patients (78%) enrolled in group I and 29 patients (64%) in group II had resolution of ST segment (P-value 0.24). Twenty-one patients (47% group I) vs 23 patients (51% group II) with P-value 0.83 achieved TIMI 0 flow. Twenty-four patients (53% group I) compared with 22 patients (49% group II) with P-value 0.83 had TIMI 1 to 3 flow before PCI. TIMI 3 flow was achieved in 40 patients (89% group I) compared with 38 patients (84% group II) with P-value 0.76. In this study there was a trend toward better ST segment resolution and patency of IRA (i.e., improved TIMI flow) in patients given high bolus dose Aggrastat in the ED. Larger studies are needed to confirm this finding.


A Randomized Control Trial Study to Determine the Effect of Melatonin on Serum Levels of IL-1β and TNF-α in Patients with Multiple Sclerosis.
Multiple sclerosis (MS) is the most common neurological disease that happens at a young age. MS is an inflammatory disease; associated with the demyelination of the central nervous system. Therefore, some inflammatory factors are effective in the mechanism and progression of the disease. Melatonin, as a multi-effect substance including anti-inflammatory effects, can reduce symptoms of MS in patients with a change in their inflammatory factors level. In this study, 50 MS patients who were referred to the MS Society of Markazi Province were randomly selected. All patients were treated with routine MS treatment (interferon) and were divided into control (25 placebo recipients) and treatment (25 recipients of 3 mg melatonin per day for 24 weeks) groups. Anthropometric data of patients including height, weight, and age were determined. Blood samples were collected after fasting in order to determine serum levels of interleukin 1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α). Then, samples were immediately centrifuged for serum separation and sera were transferred to a freezer at -80°C and serum levels of these factors were determined; using ELISA kit. The results of this study showed that there was no significant difference between the control and treatment groups in terms of serum levels of TNF-α. However, the level of IL-1β was significantly reduced in the treatment group compared to the control group, indicating that melatonin decreases this inflammatory substance. Our findings suggest a valuable strategy in the treatment of patients who suffer from MS.


A Study on the Role of Mobile Phone Communication in Tuberculosis DOTS Treatment.
Every year, a lot of Tuberculosis (TB) patients undergo Directly Observed Treatment Short-course (DOTS) in Salem city, one of the high TB districts in South India. Mobile phone usage among these patients and health workers is common. Mobile phone communication has a great potential in TB treatment. To analyze the mobile phone usage and its effectiveness in TB DOTS treatment. A cross-sectional survey with 150 TB patients was followed by a focus group discussion with treatment supervisors, DOTS providers, and health workers. Majority of patients use mobile phones to make calls to health workers to clarify their doubts on side effects, food, and symptoms of the disease. TB treatment supervisors effectively use mobile phones to counsel patients to adhere to the treatment regimen. Patients see mobile phones as a useful communication tool in TB treatment though they prefer direct interpersonal communication with health workers. Though the mobile ownership is 68% among the TB patients, many of them are not able to send text messages or read messages in English. Mobile phone possession and usage is high among the patients. Patients need to be trained to use mobile phone features such as alarm, voice mail, and interactive voice response. Incentives like free talk time and short message service (SMS) will encourage patients to communicate frequently with health workers, thereby, increasing the chances of better adherence to DOTS. SMS could be made available in the regional languages.


A curated database reveals trends in single-cell transcriptomics.
The more than 1000 single-cell transcriptomics studies that have been published to date constitute a valuable and vast resource for biological discovery. While various 'atlas' projects have collated some of the associated datasets, most questions related to specific tissue types, species or other attributes of studies require identifying papers through manual and challenging literature search. To facilitate discovery with published single-cell transcriptomics data, we have assembled a near exhaustive, manually curated database of single-cell transcriptomics studies with key information: descriptions of the type of data and technologies used, along with descriptors of the biological systems studied. Additionally, the database contains summarized information about analysis in the papers, allowing for analysis of trends in the field. As an example, we show that the number of cell types identified in scRNA-seq studies is proportional to the number of cells analysed. Database URL: www.nxn.se/single-cell-studies/gui.


Fetoscopic laser photocoagulation in the management of twin-twin transfusion syndrome: local experience from Hong Kong.
To review the perinatal outcome of monochorionic twin pregnancies treated by fetoscopic laser coagulation for twin-twin transfusion syndrome. Retrospective study. A university teaching hospital in Hong Kong. Thirty consecutive cases of fetoscopic laser coagulation of placental anastomoses for twin-twin transfusion syndrome performed in a single centre. Operative complications and perinatal survival rates. The median gestational age at initial presentation, laser photocoagulation, and delivery were 22 (range, 16-27) weeks, 23 (18-28) weeks, and 32 (21-37) weeks, respectively. To improve the visualisation, in three cases amnio-exchange was undertaken; the procedure was abandoned in two due to poor visualisation. The overall fetal survival rate, the double infant survival rate, and survival rate for at least one twin were 72% (43/60), 60% (18/30), and 83% (25/30), respectively. The most common peri-operative complication was bleeding from the uterine wall into the amniotic cavity, which affected three (10%) patients. Our results of fetoscopic laser surgery for twin-twin transfusion syndrome were similar to those in specialised centres in other countries.


Only severe COPD is associated with being underweight: results from a population survey.
Low body mass index (BMI) and malnutrition in chronic obstructive pulmonary disease (COPD) are associated with a poor prognosis. The prevalence of underweight, as well as overweight, in severity grades of COPD is sparsely investigated in studies of the general population and the associated patterns of risk factors are not well established. The aim of the present study was to determine the association between severity grades of airflow limitation in COPD, and both underweight and obesity when corrected for possible confounding factors. The study is based on pooled data from the OLIN (Obstructive Lung Disease in Northern Sweden) studies. Complete records with lung function, BMI and structured interview data were available from 3942 subjects (50.7% women and 49.3% men). COPD and severity grading were defined using the Global Initiative for Chronic Obstructive Lung Disease criteria. In sensitivity analyses, the lower limit of normal was used. The prevalence of underweight was 7.3% in severe COPD (grades 3 and 4) versus 2.0% in those with normal spirometry. The prevalence of obesity increased from 9.7% in grade 1, to 16.3% in grade 2 and 20.0% in severe COPD, versus 17.7% in those with normal spirometry. In adjusted analysis, of the COPD severity grades, only severe COPD was associated with underweight (OR 3.24, 95% CI 1.0004-10.5), while the COPD severity grades tended to be inversely associated with overweight.


Atrial fibrillation in heart failure: what should we do?
Heart failure (HF) and atrial fibrillation (AF) are two conditions that are likely to dominate the next 50 years of cardiovascular (CV) care. Both are increasingly prevalent and associated with high morbidity, mortality, and healthcare cost. They are closely inter-related with similar risk factors and shared pathophysiology. Patients with concomitant HF and AF suffer from even worse symptoms and poorer prognosis, yet evidence-based evaluation and management of this group of patients is lacking. In this review, we evaluate the common mechanisms for the development of AF in HF patients and vice versa, focusing on the evidence for potential treatment strategies. Recent data have suggested that these patients may respond differently than those with HF or AF alone. These results highlight the clear clinical need to identify and treat according to best evidence, in order to prevent adverse outcomes and reduce the huge burden that HF and AF are expected to have on global healthcare systems in the future. We propose an easy-to-use clinical mnemonic to aid the initial management of newly discovered concomitant HF and AF, the CAN-TREAT HFrEF + AF algorithm (Cardioversion if compromised; Anticoagulation unless contraindication; Normalize fluid balance; Target initial heart rate <110 b.p.m.; Renin-angiotensin-aldosterone modification; Early consideration of rhythm control; Advanced HF therapies; Treatment of other CV disease).


Thyroid Therapy or Dysfunction in Athletes: Is it Time to Revisit the Clinical Practice Guidelines?
Recent media have highlighted the controversy surrounding treatment of elite athletes for hypothyroidism. The World Anti-Doping Agency denied a request by the United States Anti-Doping Agency to ban the use of thyroid medication. At present, there is no scientific evidence that thyroid medication has the potential to enhance performance. Clinical practice guidelines are not definitive in regard to what classifies a patient as having hypothyroidism. Thyroid-stimulating hormone and free T4 are recommended to screen for thyroid disease; however, the thyrotropin-releasing hormone stimulation test is still advocated by some for detecting the earliest stages of hypothyroidism. Hypothyroidism has been demonstrated to reduce cardiopulmonary function and result in musculoskeletal symptoms, such as fatigue and muscle stiffness. Symptoms of hypothyroidism, including depression, fatigue, and impaired sleep, are similar to those reported in overtraining. These patients may have hypothalamic-pituitary dysfunction that may complicate interpretation of basal thyroid-stimulating hormone and free T4. To date, no association has been identified between training state and hypothyroidism. Research to more clearly define hypothyroidism using provocative testing, evaluate the potential for thyroid medication to enhance performance, and examine whether training may induce hypothyroidism in athletes is desirable.


Gliclazide produces high-affinity block of KATP channels in mouse isolated pancreatic beta cells but not rat heart or arterial smooth muscle cells.
Sulphonylureas stimulate insulin secretion by closing ATP-sensitive potassium (KATP) channels in the pancreatic beta-cell membrane. KATP channels are also found in other tissues, including heart and smooth muscle, where they link cellular metabolism to electrical activity. The sulphonylurea gliclazide blocks recombinant beta-cell KATP channels (Kir6.2/SUR1) but not heart (Kir6.2/SUR2A) or smooth muscle (Kir6.2/SUR2B) KATP channels with high potency. In this study, we examined the specificity of gliclazide for the native (as opposed to recombinant) KATP channels in beta cells, heart and smooth muscle. The action of the drug was studied by whole-cell current recordings of native KATP channels in isolated pancreatic beta-cells and myocytes from heart and smooth muscle. Gliclazide blocked whole-cell beta-cell KATP currents with an IC50 of 184 +/- 30 nmol/l (n = 6-10) but was much less effective in cardiac and smooth muscle (IC50s of 19.5 +/- 5.4 micromol/l (n = 6-12) and 37.9 +/- 1.0 micromol/l (n = 5-10), respectively). In all three tissues, the action of the drug on whole-cell KATP currents was rapidly reversible. In inside-out patches on beta-cells, gliclazide (1 micromol/l) produced a maximum of 66 +/- 13 % inhibition (n = 5), compared with more than 98 % block in the whole-cell configuration. Gliclazide is a high-potency sulphonylurea which shows specificity for the pancreatic beta-cell KATP channel over heart and smooth muscle. In this respect, it differs from glibenclamide. The difference in the maximal block observed in the excised patch and whole-cell recordings from beta-cells, may be due to the absence of intracellular Mg-nucleotides in the excised patch experiments.


Epidermal cell proliferation in calorie-restricted aging rats.
Calorie restriction (CR) has been known to produce many beneficial health effects, and lowered cell proliferation from CR has been shown to produce anti-cancer effects in some tissues. In this study the rate of epidermal cell proliferation in aging Fischer 344 rats from ad libitum fed (AL) and CR colonies was assessed in relation to changes in epidermal thickness with age. Proliferating cell nuclear antigen (PCNA) was detected using immunohistochemical method on paraffin sections in the epidermis of dorsal skin and footpad in these animals obtained from the National Institute on Aging. The proliferating cell index was compared with morphometric measurement of epidermis in young, young adult and old animals (six per group). Data were analyzed by Excel and SPSS 14.0 softwares for statistical evaluation. Two-way analysis of variance (ANOVA) was applied to data to test the effects of age, diet, and age-diet interaction. The following significant effects were noted: (I) age and age-diet effects in dorsal skin epidermal width, and PCNA; (II) age, and diet effect on footpad epidermal thickness, and PCNA index. There was a trend of increasing epidermal thickness in the dorsal skin in normally feeding aging rats which was depressed with CR in the two younger groups. PCNA index showed a trend of attrition from young to old. The thickness of epidermis in foot pad showed a curvilinear trend in both AL and CR groups with lowest mean values in the old group, and more predominant effect in CR-exposed animals. The proliferation index in the foot pad demonstrated a trend of reduction in old specimens with lower mean values in each corresponding CR age group. This report agrees with CR-inhibited cell proliferation reported in many organs by other investigators, and the observed results might have been caused by physiological or endocrine mechanisms affecting the epithelium in these calorie-restricted animals.


Neuropsychiatric symptoms are common in immunocompetent adult patients with Toxoplasma gondii acute lymphadenitis.
Chronic toxoplasmosis has been shown to be strongly associated with a range of neuropsychiatric effects including schizophrenia and suicide. However there have not been any prospective, community-based studies of the neuropsychiatric effects of acute toxoplasmosis in adult immunocompetent patients. Adult patients with a positive serum IgM anti-Toxoplasma gondii test result, in the context of an acute illness with lymphadenopathy, were invited to complete a questionnaire seeking information relating to the nature, severity, and duration of symptoms in the months following the diagnosis of acute toxoplasmosis. Laboratory testing identified a total of 187 adults who had a positive serum IgM anti-T. gondii test result between 1 January and 30 November 2011. Consent to contact 108/187 (58%) patients was provided by their family doctor; 37 (34%) of these 108 patients completed and returned the questionnaire. Questionnaires from the 31/108 (29%) patients who reported swollen lymph nodes during their illness were included in the study. Fatigue (90%), headache (74%), difficulty concentrating (52%), and muscle aches (52%) were the most commonly reported symptoms. These symptoms commonly persisted for at least 4 weeks. Twenty-seven of 31 (87%) subjects reported a moderate or severe reduction in their overall physical and mental health during the first 2 months of illness. Acute toxoplasmosis in immunocompetent adults commonly causes moderately severe neuropsychiatric symptoms that might result from replication of the organism in the central nervous system with consequent effects on brain function. Patients should be advised that such symptoms are common and reassured that they usually resolve completely within a few months.


Snacktivity™ to promote physical activity and reduce future risk of disease in the population: protocol for a feasibility randomised controlled trial and nested qualitative study.
Many people do not regularly participate in physical activity, which may negatively impact their health. Current physical activity guidelines are focused on promoting weekly accumulation of at least 150 min of moderate to vigorous intensity physical activity (MVPA). Whilst revised guidance now recognises the importance of making small changes to physical activity behaviour, guidance still focuses on adults needing to achieve at least 150 min of MVPA per week. An alternative 'whole day' approach that could motivate the public to be more physically active, is a concept called Snacktivity™. Instead of focusing on achieving 150 min per week of physical activity, for example 30 min of MVPA over 5 days, Snacktivity™ encourages the public to achieve this through small, but frequent, 2-5 min 'snacks' of MVPA throughout the whole day. The primary aim is to undertake a feasibility trial with nested qualitative interviews to assess the feasibility and acceptability of the Snacktivity™ intervention to inform the design of a subsequent phase III randomised trial. A two-arm randomised controlled feasibility trial aiming to recruit 80 inactive adults will be conducted. Recruitment will be from health and community settings and social media. Participants will be individually randomised (1:1 ratio) to receive either the Snacktivity™ intervention or usual care. The intervention will last 12 weeks with assessment of outcomes completed before and after the intervention in all participants. We are interested in whether the Snacktivity™ trial is appealing to participants (assessed by the recruitment rate) and if the Snacktivity™ intervention and trial methods are acceptable to participants (assessed by Snacktivity™/physical activity adherence and retention rates). The intervention will be delivered by health care providers within health care consultations or by researchers. Participants' experiences of the trial and intervention, and health care providers' views of delivering the intervention within health consultations will be explored. The development of physical activity interventions that can be delivered at scale are needed. The findings from this study will inform the viability and design of a phase III trial to assess the effectiveness and cost-effectiveness of Snacktivity™ to increase physical activity. ISRCTN: 64851242.


Neurofilament light chain in serum is significantly increased in chorea-acanthocytosis.
Chorea-acanthocytosis (ChAc) is a rare hereditary neurodegenerative disease, characterized by hyper- and hypokinetic movement disorders, peripheral neuropathy and acanthocytosis. Biomarkers are not established; possible candidates include neurofilament reflecting neuroaxonal damage. We studied serum neurofilament light chain (sNfL) of six ChAc patients compared to two healthy control cohorts (A, six age/sex matched and B, historical cohort of 59 healthy adult subjects) and in two patients with the very similar condition of McLeod syndrome (MLS), the second core syndrome of neuroacanthocytosis. sNfL was quantified using single-molecule array analysis. sNfL concentration was significantly higher in the ChAc cohort (18.73 pg/ml; IQR 15.65-27.70) compared to both healthy control cohorts (A, 7.37 pg/ml; IQR 5.60-9.05; B, 3.10 pg/ml; IQR 2.43-3.98). In MLS patients, a similar sNfL increase was observed. sNfL is significantly increased in ChAc and MLS and seems to reflect neuroaxonal damage in the peripheral as well as the central nervous system.


Origin and specificity of autoreactive T cells in antigen-induced populations.
We have characterized the major histocompatibility complex (MHC) specificity of autoreactive T cell clones arising from diverse donors after immunization with different antigens. The MHC fine specificity of autoreactive T cells for unique F1 hybrid determinants of BALB.K X BALB.B F1, and for the mutant I-Ab determinants of the B6.C-H-2bm12 (bm 12) strain is similar to that previously described for antigen-specific T cells. We find, furthermore, that the MHC specificity of autoreactive T cell clones selected from primed populations grown in the absence of Con A-stimulated supernatant factors reflects the predominant MHC restriction specificity of T cells specific for the immunogen. Thus, I-E subregion-specific autoreactive T cells are detected at a much higher frequency after immunization with the I-E-restricted antigen, GL (terpolymer of glutamic acid, lysine, and phenylalanine), than with the predominantly I-A-restricted antigen, keyhole limpet hemocyanin (KLH). These experiments strongly suggest that some autoreactive T cells are derived from antigen-stimulated precursors. This result contrasts with that obtained when autoreactive T cells are selected in bulk cultures, or in the presence of exogenous T cell factors. We conclude that, under optimal conditions, most autoreactive T cells are recruited from a relatively stable pool of predominantly I-A-specific precursors. Autoreactive precursors in this pool might themselves derive from previous antigenic stimulation, or be of independent origin.


Hydrated nucleus pulposus extrusion in dogs: correlation of magnetic resonance imaging and microsurgical findings.
Magnetic resonance imaging (MRI) patterns of canine cervical hydrated nucleus pulposus extrusion (HNPE) have been described by a few reports, but the correlation between microsurgical and MRI features has never been investigated. The aim of this study was to compare the MRI features of HNPE with microsurgical findings and cytological outcomes and also to investigate the anatomical and pathophysiological aspects of the disease. A prospective clinical study was conducted in 36 dogs suffering from HNPE. The diagnosis was based on high-field MRI findings of ventral extradural lesions, adjacent to the dorsal aspect of intervertebral discs, characterised by high signal intensity in T2-weighted sequences and hypointensity in T1-weighted sequences. MRI images were analysed with regard to the intervertebral space involved, the grading of spinal cord compression, the signal intensity and distribution of the material, and the thickness and signal intensity of the involved discs. All patients underwent microsurgical decompression and direct observations were recorded and films of the surgical procedure analysed. The majority of patients had acute onset of clinical signs (78 %), the patient did not exhibit signs of pain in 75 % of dogs and neurological deficits varied from slight tetraparesis (33 %) to tetraplegia (28 %). The localization of the extruded disc material was ventral relative to the dorsal longitudinal ligament that was lifted dorsally and appeared intact at the site of compression. Direct microsurgical observations of the HNPE sites showed that extruded disc material was collected within the fibres of the dorsal longitudinal ligament. The consistency was gelatinous in 42 %, water-like in 33 %, and lumpy liquid in 25 % of cases. Cytological samples did not detect the presence of inflammation, bacteria, fungi, neoplastic cells or foreign material. Microsurgical features of HNPE suggest that the extruded disc is collected within the fibres of the dorsal longitudinal ligament and this may explain the typical MRI appearance of this disease. Further pathophysiological studies are needed to investigate why the cervical nucleus pulposus extrusion appears to occur without obvious trauma.


Toll-like receptor 4: a novel signaling pathway during renal fibrogenesis.
The toll-like receptor (TLR) family serves an important regulatory role in the innate immune system, and recent evidence has implicated TLR signaling in the pro-inflammatory response of a variety of endogenous and exogenous stimuli within the kidney. The role of TLR signaling in fibrotic renal injury, however, remains unknown. C3H/HeJ TLR4 hyporesponsive mice (TLR4(Lps-d)) or WT controls (C3H/HeOu/J) underwent either sham operation or 1 wk of unilateral ureteral obstruction (UUO). The kidneys were harvested and tissues were analyzed for TLR4 expression (Western blot; RTPCR), E-cadherin and alpha smooth muscle actin (α-SMA) expression (Western blot), fibroblast accumulation (fibroblast specific protein (FSP-1+) staining), renal fibrosis (collagen I RTPCR, total collagen assay, Masson's trichrome staining), cytokine gene expression (tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta1 (TGF-β1) RTPCR), and pSMAD2 and integrin α1 expression (Western blot). Mice with intact TLR4 signaling demonstrate a significant increase in TLR4 expression, α-SMA expression, fibroblast accumulation, collagen deposition, and interstitial fibrosis, and a significant decrease in E-cadherin expression in response to UUO. TLR4 deficient mice, however, exhibit a significant reduction in obstruction-induced α-SMA expression, fibroblast accumulation, and renal fibrosis, with preservation of E-cadherin expression. TLR4's influence on fibroblast accumulation and renal fibrosis occurred independent of any alterations in TNF-α, TGF-β1, or pSMAD2 expression, but did involve alterations integrin α1 expression. TLR4 appears to be a significant mediator of fibrotic renal injury. While TLR4 signaling is recognized as a critical component of the innate immune response, this is the first study to demonstrate a novel role for TLR4 in renal fibroblast accumulation and tubulointerstitial fibrosis.


Activity of human motor system during action observation is modulated by object presence.
Neurons in the monkey mirror neuron system (MNS) become active when actions are observed or executed. Increases in activity are greater when objects are engaged than when the actions are mimed. This modulation occurs even when object manipulation is hidden from view. We examined whether human motor systems are similarly modulated during action observation because such observation-related modulations are potentially mediated by a putative human MNS. Transcranial magnetic stimulation (TMS) was used to elicit motor-evoked potentials (MEPs) of a grasping muscle while participants observed actual or pantomimed grasping movements whose endpoints were sometimes hidden from view. MEP amplitudes were found to be modulated by object presence. Critically, the object-based modulation was found when the participant directly observed object manipulation and when the object manipulation had to be inferred because it was hidden. These findings parallel studies of MNS activity in monkeys and support the hypothesis that the MNS influences motor system activity during action observation. Although the object-based modulation of MEP amplitudes was consistent with the hypotheses, the direction of the modulation was not--MEP amplitudes decreased during action observation in contrast to the increase that has previously been observed. We suggest that the decrease in MEP amplitude on object-present trials resulted from inhibitory mechanisms that were activated to suppress the observation-evoked response codes from generating overt muscle activity.


The role of pemetrexed in recurrent epithelial ovarian cancer: A scoping review.
Ovarian cancer is the leading cause of mortality among gynecologic malignancies, with most cases diagnosed at an advanced stage. Despite an initial response, most develop a recurrence and subsequent resistance to standard therapies. Pemetrexed (AlimtaTM) is a new generation multi-targeted antifolate initially approved for the treatment of malignant pleural mesothelioma. In recent years, it has shown promise in the treatment of recurrent epithelial ovarian cancer. In this review, we outline the current literature and discuss the future of pemetrexed in the setting of recurrent epithelial ovarian cancer.


Neural dynamics of sentiment processing during naturalistic sentence reading.
When we read, our eyes move through the text in a series of fixations and high-velocity saccades to extract visual information. This process allows the brain to obtain meaning, e.g., about sentiment, or the emotional valence, expressed in the written text. How exactly the brain extracts the sentiment of single words during naturalistic reading is largely unknown. This is due to the challenges of naturalistic imaging, which has previously led researchers to employ highly controlled, timed word-by-word presentations of custom reading materials that lack ecological validity. Here, we aimed to assess the electrical neural correlates of word sentiment processing during naturalistic reading of English sentences. We used a publicly available dataset of simultaneous electroencephalography (EEG), eye-tracking recordings, and word-level semantic annotations from 7129 words in 400 sentences (Zurich Cognitive Language Processing Corpus; Hollenstein et al., 2018). We computed fixation-related potentials (FRPs), which are evoked electrical responses time-locked to the onset of fixations. A general linear mixed model analysis of FRPs cleaned from visual- and motor-evoked activity showed a topographical difference between the positive and negative sentiment condition in the 224-304 ms interval after fixation onset in left-central and right-posterior electrode clusters. An additional analysis that included word-, phrase-, and sentence-level sentiment predictors showed the same FRP differences for the word-level sentiment, but no additional FRP differences for phrase- and sentence-level sentiment. Furthermore, decoding analysis that classified word sentiment (positive or negative) from sentiment-matched 40-trial average FRPs showed a 0.60 average accuracy (95% confidence interval: [0.58, 0.61]). Control analyses ruled out that these results were based on differences in eye movements or linguistic features other than word sentiment. Our results extend previous research by showing that the emotional valence of lexico-semantic stimuli evoke a fast electrical neural response upon word fixation during naturalistic reading. These results provide an important step to identify the neural processes of lexico-semantic processing in ecologically valid conditions and can serve to improve computer algorithms for natural language processing.


Design and synthesis of cenocladamide analogues and their evaluation against breast cancer cell lines.
This work describes the total synthesis of the alkaloid cenocladamide and a concise library of nine structural analogues aiming at their evaluation against the breast cancer cell line MDA-MB-231. The most promising compound (3; IC50 = 6.6 μM) was also evaluated in a panel of seven breast cancer cell lines and two non-tumorigenic cell lines. We further conducted an initial investigation on the mechanism of action of analogue 3, which lacks the endocyclic double bond when compared to cenocladamide. The present study presents the discovery of a cenocladamide analogue with interesting cytotoxic activity, which could be useful for further optimization towards new chemotherapeutic agents for breast cancer treatment.


Inhibition of human recombinant T-type calcium channels by the endocannabinoid N-arachidonoyl dopamine.
N-arachidonoyl dopamine (NADA) has complex effects on nociception mediated via cannabinoid CB(1) receptors and the transient receptor potential vanilloid receptor 1 (TRPV1). Anandamide, the prototypic CB(1)/TRPV1 agonist, also inhibits T-type voltage-gated calcium channel currents (I(Ca)). These channels are expressed by many excitable cells, including neurons involved in pain detection and processing. We sought to determine whether NADA and the prototypic arachidonoyl amino acid, N-arachidonoyl glycine (NAGly) modulate T-type I(Ca) Human recombinant T-type I(Ca) (Ca(V)3 channels) expressed in HEK 293 cells and native mouse T-type I(Ca) were examined using standard whole-cell voltage clamp electrophysiology techniques. N-arachidonoyl dopamine completely inhibited Ca(V)3 channels with a rank order of potency (pEC(50)) of Ca(V)3.3 (6.45) > or = Ca(V)3.1 (6.29) > Ca(V)3.2 (5.95). NAGly (10 micromol.L(-1)) inhibited Ca(V)3 I(Ca) by approximately 50% or less. The effects of NADA and NAGly were voltage- but not use-dependent, and both compounds produced significant hyperpolarizing shifts in Ca(V)3 channel steady-state inactivation relationships. By contrast with anandamide, NADA and NAGly had modest effects on Ca(V)3 channel kinetics. Both NAGly and NADA inhibited native T-type I(Ca) in mouse sensory neurons. N-arachidonoyl dopamine and NAGly increase the steady-state inactivation of Ca(V)3 channels, reducing the number of channels available to open during depolarization. These effects occur at NADA concentrations at or below to those affecting CB(1) and TRPV1 receptors. Together with anandamide, the arachidonoyl neurotransmitter amides, NADA and NAGly, represent a new family of endogenous T-type I(Ca) modulators.


Calculating free energies from the vibrational density of states function: Validation and critical assessment.
We explore and show the usefulness of the density of states function for computing vibrational free energies and free energy differences between small systems. Therefore, we compare this density of states integration method (DSI) to more established schemes such as Bennett's Acceptance Ratio method (BAR), the Normal Mode Analysis (NMA), and the Quasiharmonic Analysis (QHA). The strengths and shortcomings of all methods are highlighted with three numerical examples. Furthermore, the free energy of the ionization of ammonia and the mutation from serine to cysteine are computed using extensive ab initio molecular dynamics simulations. We conclude that DSI improves upon the other frequency-based methods (NMA and QHA) regarding the treatment of anharmonicity and yielding results comparable to BAR in all cases without the need for alchemical transformations. Low-frequency modes lead to larger errors indicating that long simulation times might be required for larger systems. In addition, we introduce the use of DSI for the localization of the vibrational free energy to specific atoms or residues, leading to insights into the underlying process, a unique feature that is only offered by this method.


Frontal Hemodynamic Response During Step Initiation Under Cognitive Conflict in Older and Young Healthy People.
Gait initiation is a daily challenge even for healthy individuals as it requires the timely coupling between the automatic anticipatory postural adjustment (APA) and the voluntary step according to the context. Modulation of this motor event has been thought to involve higher level brain control, including cognitive inhibitory circuitries. Despite the known participation of the supplementary motor area (SMA) in the modulation of some parameters of APA, the participation of areas controlling inhibition during gait initiation still needs to be investigated. In this study, the hemodynamic responses of the SMA and dorsolateral prefrontal cortex (DLPFC) were assessed using functional near-infrared spectroscopy (fNIRS) during a gait initiation task under cognitive conflict to select the foot to step (congruent [CON] and incongruent [INC] conditions). The older group (OG) showed worse inhibitory control than the young group (YG) along with more impairments in APA parameters. OG also had a lower amplitude of hemodynamic responses in both areas than YG in the INC. The INC increased the correlation between SMA and DLPFC only in the YG. Aging seems to impair the interaction between the hemodynamic responses of SMA and DLPFC, which influences APA performance in gait initiation under cognitive conflict.