Acute pain: a multifaceted challenge - the role of nimesulide.
This article summarizes the outcome from an international consensus meeting, which took place in Vienna on 4 November 2014. The aim of the meeting was to provide the state of the art on the pathophysiology and treatment of acute pain with special emphasis on nimesulide, a non-steroidal anti-inflammatory drug (NSAID) indicated for the treatment of acute pain and primary dysmenorrhea. Besides the data on the mechanisms of acute inflammatory pain and on the efficacy and safety of nimesulide in patients affected by different forms of acute pain, the clinical experience of attending experts was discussed based on selected case reports. The members of this consensus group recognized that nimesulide is a NSAID highly effective in the treatment of several painful situations with an acute inflammatory component including primary dysmenorrhea. Although safety concerns regarding nimesulide have emerged in recent years, both robust new epidemiological data and clinical experience confirm a positive benefit/risk profile of nimesulide in the treatment of several forms of acute pain. The members of this international consensus group concluded that nimesulide, when used appropriately, remains a particularly valuable and safe option for the treatment of several conditions characterized by the presence of acute inflammatory pain because of the rapid onset of the analgesic action, and the positive evidence-based benefit/risk profile.


Rapid onset opioids in palliative medicine.
Opioids are potent broad-spectrum analgesics useful to treat a variety of painful conditions. Opioids have been classified by many different categorizations (e.g. natural, semisynthetic, synthetic). One classification that may be useful in terms of selecting specific opioid agents is based on the pharmacokinetic characteristics of the various opioid formulations (e.g. longacting opioids, short-acting opioids, and rapid onset opioids). These various opioid agents appear to be useful to "match" specific types of pains. Rapid onset opioids appear especially well-suited to treat breakthrough pain and in particular, rapidonset breakthrough pain. This article will briefly review 5 rapid onset opioids that are FDA approved in the U.S.


Genome-wide differentiation in closely related populations: the roles of selection and geographic isolation.
Population divergence in geographic isolation is due to a combination of factors. Natural and sexual selection may be important in shaping patterns of population differentiation, a pattern referred to as 'isolation by adaptation' (IBA). IBA can be complementary to the well-known pattern of 'isolation by distance' (IBD), in which the divergence of closely related populations (via any evolutionary process) is associated with geographic isolation. The barn swallow Hirundo rustica complex comprises six closely related subspecies, where divergent sexual selection is associated with phenotypic differentiation among allopatric populations. To investigate the relative contributions of selection and geographic distance to genome-wide differentiation, we compared genotypic and phenotypic variation from 350 barn swallows sampled across eight populations (28 pairwise comparisons) from four different subspecies. We report a draft whole-genome sequence for H. rustica, to which we aligned a set of 9493 single nucleotide polymorphisms (SNPs). Using statistical approaches to control for spatial autocorrelation of phenotypic variables and geographic distance, we find that divergence in traits related to migratory behaviour and sexual signalling, as well as geographic distance, together explain over 70% of genome-wide divergence among populations. Controlling for IBD, we find 42% of genomewide divergence is attributable to IBA through pairwise differences in traits related to migratory behaviour and sexual signalling alone. By (i) combining these results with prior studies of how selection shapes morphological differentiation and (ii) accounting for spatial autocorrelation, we infer that morphological adaptation plays a large role in shaping population-level differentiation in this group of closely related populations.


The clinical and prognostic value of motor abnormalities in psychosis, and the importance of instrumental assessment.
Motor abnormalities comprise several clinical signs intrinsic to psychosis. Critically, these features are of prognostic value in individuals at-risk for psychosis, and for those in early stages of psychotic disorders. Motor abnormalities such as tremor, rigidity, and neurological soft signs often go unrecognized. Currently, advances in this area are limited by a paucity of theoretical conceptions categorizing or linking these behaviours to underlying neurobiology affected in psychosis. However, emerging technological advances have significantly improved the ability to detect and assess motor abnormalities with objective instruments in a timely and reliable manner. Further, converging evidence has laid the groundwork for theoretically and empirically derived categorization and conceptualization. This review summarizes these advances, stressing the importance of motor abnormalities for understanding vulnerability across different stages of psychosis and introducing these innovative instrumental approaches. Patients, researchers and clinicians will benefit from these new developments, as better assessment aids the development of targeted interventions to ultimately improve the care for individuals experiencing psychosis.


Validation of a nomogram to predict disease progression following salvage radiotherapy after radical prostatectomy: results from the SEARCH database.
To externally validate the nomogram published by Stephenson et al. (termed the 'Stephenson nomogram') to predict disease progression after salvage radiotherapy (SRT) among patients with prostate cancer from the Shared Equal Access Regional Cancer Hospital (SEARCH) database. We analysed data from 102 men treated with SRT for prostate-specific antigen (PSA) failure after prostatectomy, of whom 30 (29%) developed disease progression after SRT during a median follow-up of 50 months. The predicted 6-year progression-free survival (PFS) was compared to the actuarial PFS using calibration plots. The accuracy of the nomogram to risk-stratify men for progression was assessed by the concordance index. The median PSA and PSA doubling time before SRT was 0.6 ng/mL and 10.3 months, respectively. The 6-year actuarial disease-free progression after SRT was 57% (95% confidence interval 42-69%). The overall concordance index of the Stephenson nomogram was 0.65. The nomogram predicted failure more accurately at the extremes of risk (lowest and highest) but in intermediate groups, the accuracy was less precise. Of the 11 variables used in the nomogram, only negative margins and high PSA level before SRT were significantly associated with increased disease progression. The Stephenson nomogram is an important tool to predict disease progression after SRT following radical prostatectomy. It adequately predicted progression in SEARCH with reasonable accuracy. Also, in SEARCH, disease progression was predicted by similar disease characteristics. However, the overall modest performance of the model in our validation cohort indicates there is still room for improvement in predictive models for disease progression after SRT.


Case Study: Prolonged Infectious SARS-CoV-2 Shedding from an Asymptomatic Immunocompromised Individual with Cancer.
Long-term severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) shedding was observed from the upper respiratory tract of a female immunocompromised individual with chronic lymphocytic leukemia and acquired hypogammaglobulinemia. Shedding of infectious SARS-CoV-2 was observed up to 70 days, and of genomic and subgenomic RNA up to 105 days, after initial diagnosis. The infection was not cleared after the first treatment with convalescent plasma, suggesting a limited effect on SARS-CoV-2 in the upper respiratory tract of this individual. Several weeks after a second convalescent plasma transfusion, SARS-CoV-2 RNA was no longer detected. We observed marked within-host genomic evolution of SARS-CoV-2 with continuous turnover of dominant viral variants. However, replication kinetics in Vero E6 cells and primary human alveolar epithelial tissues were not affected. Our data indicate that certain immunocompromised individuals may shed infectious virus longer than previously recognized. Detection of subgenomic RNA is recommended in persistently SARS-CoV-2-positive individuals as a proxy for shedding of infectious virus.


Combining fish-oil supplements with regular aerobic exercise improves body composition and cardiovascular disease risk factors.
Regular exercise and consuming long-chain n-3 fatty acids (FAs) from fish or fish oil can independently improve cardiovascular and metabolic health, but combining these lifestyle modifications may be more effective than either treatment alone. We examined the individual and combined effects of n-3 FA supplements and regular exercise on body composition and cardiovascular health. Overweight volunteers [body mass index (BMI; in kg/m(2)): >25] with high blood pressure, cholesterol, or triacylglycerols were randomly assigned to one of the following interventions: fish oil (FO), FO and exercise (FOX), sunflower oil (SO; control), or SO and exercise (SOX). Subjects consumed 6 g tuna FO/d ( approximately 1.9 g n-3 FA) or 6 g SO/d. The exercise groups walked 3 d/wk for 45 min at 75% age-predicted maximal heart rate. Plasma lipids, blood pressure, and arterial function were assessed at 0, 6, and 12 wk. Body composition was assessed by dual-energy X-ray absorptiometry at 0 and 12 wk only. FO supplementation lowered triacylglycerols, increased HDL cholesterol, and improved endothelium-dependent arterial vasodilation (P<0.05). Exercise improved arterial compliance (P<0.05). Both fish oil and exercise independently reduced body fat (P<0.05). FO supplements and regular exercise both reduce body fat and improve cardiovascular and metabolic health. Increasing intake of n-3 FAs could be a useful adjunct to exercise programs aimed at improving body composition and decreasing cardiovascular disease risk.


Long-term neuropsychological outcome in preterm twins.
Few long-term studies have yet described neuropsychological outcome in preterm twins. Our aim was to assess, by long-term evaluation, neuropsychological outcome in preterm twins in order to define a correct follow-up program. Our study was a cohort one, with an index and a comparison group. Neonatal medical records of all preterm newborns admitted to our centre between 1991 and 1997 were reviewed and selected patients were recalled. The sample population included two matched groups of children aged 6-12 years, 86 twins and 86 singletons, submitted to paediatric, neurological, psychological, and ophthalmological examinations. Inclusion criteria were twin pregnancy and gestational age 27-36 weeks for index group; same gestational age, but single pregnancy, for the comparison group. All children underwent paediatric and neuropsychiatric examinations, cognitive assessment, and psychological evaluation by standardized tests for screening of learning specific disorders and language difficulties, and finally, ophthalmological examination. In order to study their role in predicting neuropsychological outcome, we examined some perinatal prognostic factors by statistical analysis. Unfavourable neuropsychological outcome was observed in 55/172 (32%) children, with different prevalence in the two groups, 42/172 (24%) in twins and 13/172 (8%) in singletons. Statistical analysis performed for examined prognostic factors showed significant differences in neuropsychological outcome with regard only to gestational age < 32 weeks, low birth weight, intraventricular haemorrhage, and periventricular leukomalacia. The incidence of neuropsychological diseases in the two groups showed significant difference about language and learning difficulties. Our data suggest that preterm twins represent a particular high-risk category of premature babies, mostly regarding the risk of so-called "minimal brain dysfunction", so a careful follow-up is recommended.


The effect of testosterone replacement therapy on prostate cancer: a systematic review and meta-analysis.
Testosterone replacement therapy (TRT) is a widely accepted form of treatment worldwide for aging men with late-onset hypogonadism syndrome. Urologists have been concerned about the possibility of TRT causing prostate cancer. The aim of this study was to assess the relationship between TRT and prostate cancer. A literature review was performed to identify all published, randomized controlled trials (RCTs) of testosterone treatment for hypogonadism. The search included the MEDLINE, Embase and the Cochrane Controlled Trials Register databases. Fixed-effect model was chosen for homogeneous studies; otherwise, a random-effect model was used. Inconsistency was quantified by using the I2 statistic, which tests the proportion of heterogeneity across studies. Results of 22 RCTs involving a total of 2351 patients were analyzed. Eleven RCTs were short-term (<12 months) and 11 were long-term (12-36 months) comparisons of TRT with a placebo; TRT was administered transdermally, orally or by injection. Respective odds ratio (OR) and 95% confidence interval (CI) values for injection, transdermal administration and oral administration of short-term TRT were as follows: prostate cancer: 0.39 (0.06-2.45), 1.10 (0.26-4.65) and no oral; biopsy: 5.28 (0.24-113.87), 2.11 (0.32-13.73) and no oral; and prostate nodule: 1.01 (0.13-7.60), no injection and oral. Respective OR and 95% CI values for injection, transdermal administration and oral administration of long-term TRT were as follows: prostate cancer: 2.09 (0.18-24.73), 3.06 (0.12-76.70) and 0.19 (0.01-4.03); biopsy: 2.09 (0.18-24.73), 3.65 (0.88-15.20) and 0.97 (0.13-7.03); and prostate nodule: 3.13 (0.12-80.68), 1.00 (0.06-16.41) and 0.97 (0.13-7.03). Though for some routes of administration and some end points, the OR associated with testosterone administration were >1 indicating increased risk, none of these reached or even approached statistical significance (all P>0.10), which was consistent with the results of subgroup analyses and sensitivity analysis. Besides, sensitivity analysis indicated that short-term TRT was more likely to increase PSA levels than treatment with placebo (P<0.00001). This meta-analysis shows that regardless of the administration method, TRT is the short-term safety and does not promote prostate cancer development or progression but long-term data are warranted with justifiable end points.


Astronomy and solar physics: measurement of the solar spectral irradiance from 200 to 3000 nanometers.
The solar spectrum experiment on Spacelab 1 measured 98 percent of the sun's total energy output. It improved the absolute accuracy of solar irradiance data, especially in the ultraviolet and infrared regions. In order to detect any variation in the spectrum on future shuttle flights, the data were obtained in a radiation scale that can be preserved with high precision over many years. The instrument performance and preliminary data reduction are described.


The multidrug resistance 1 gene Abcb1 in brain and placenta: comparative analysis in human and guinea pig.
The Multidrug Resistance 1 (MDR1; alternatively ABCB1) gene product P-glycoprotein (P-gp), an ATP binding cassette transporter, extrudes multiple endogenous and exogenous substrates from the cell, playing an important role in normal physiology and xenobiotic distribution and bioavailability. To date, the predominant animal models used to investigate the role of P-gp have been the mouse and rat, which have two distinct genes, Abcb1a and Abcb1b. In contrast, the human has a single gene, ABCB1, for which only a single isoform has been validated. We and others have previously shown important differences between Abcb1a and Abcb1b, limiting the extrapolation from rodent findings to the human. Since the guinea pig has a relatively long gestation, hemomonochorial placentation and neuroanatomically mature offspring, it is more similar to the human, and may provide a more comparable model for investigating the regulation of P-gp in the brain and placenta, however, to date, the Abcb1 gene in the guinea pig remains to be characterized. The placenta and fetal brain are barrier sites that express P-gp and that play a critical role of protection of the fetus and the fetal brain from maternally administered drugs and other xenobiotics. Using RNA sequencing (RNA-seq), reverse transcription-polymerase chain reaction (RT-PCR) and quantitative PCR (QPCR) to sequence the expressed isoforms of guinea pig Abcb1, we demonstrate that like the human, the guinea pig genome contains one gene for Abcb1 but that it is expressed as at least three different isoforms via alternative splicing and alternate exon usage. Further, we demonstrate that these isoforms are more closely related to human than to rat or mouse isoforms. This striking, overall similarity and evolutionary relatedness between guinea pig Abcb1 and human ABCB1 indicate that the guinea pig represents a relevant animal model for investigating the function and regulation of P-gp in the placenta and brain.


Network neuroscience of apathy in cerebrovascular disease.
Apathy is a reduction in motivated goal-directed behavior (GDB) that is prevalent in cerebrovascular disease, providing an important opportunity to study the mechanistic underpinnings of motivation in humans. Focal lesions, such as those seen in stroke, have been crucial in developing models of brain regions underlying motivated behavior, while studies of cerebral small vessel disease (SVD) have helped define the connections between brain regions supporting such behavior. However, current lesion-based models cannot fully explain the neurobiology of apathy in stroke and SVD. To address this, we propose a network-based model which conceptualizes apathy as the result of damage to GDB-related networks. A review of the current evidence suggests that cerebrovascular disease-related pathology can lead to network changes outside of initially damaged territories, which may propagate to regions that share structural or functional connections. The presentation and longitudinal trajectory of apathy in stroke and SVD may be the result of these network changes. Distinct subnetworks might support cognitive components of GDB, the disruption of which results in specific symptoms of apathy. This network-based model of apathy may open new approaches for investigating its underlying neurobiology, and presents novel opportunities for its diagnosis and treatment.


Diversification and niche adaptations of Nitrospina-like bacteria in the polyextreme interfaces of Red Sea brines.
Nitrite-oxidizing bacteria (NOB) of the genus Nitrospina have exclusively been found in marine environments. In the brine-seawater interface layer of Atlantis II Deep (Red Sea), Nitrospina-like bacteria constitute up to one-third of the bacterial 16S ribosomal RNA (rRNA) gene sequences. This is much higher compared with that reported in other marine habitats (~10% of all bacteria), and was unexpected because no NOB culture has been observed to grow above 4.0% salinity, presumably due to the low net energy gained from their metabolism that is insufficient for both growth and osmoregulation. Using phylogenetics, single-cell genomics and metagenomic fragment recruitment approaches, we document here that these Nitrospina-like bacteria, designated as Candidatus Nitromaritima RS, are not only highly diverged from the type species Nitrospina gracilis (pairwise genome identity of 69%) but are also ubiquitous in the deeper, highly saline interface layers (up to 11.2% salinity) with temperatures of up to 52 °C. Comparative pan-genome analyses revealed that less than half of the predicted proteome of Ca. Nitromaritima RS is shared with N. gracilis. Interestingly, the capacity for nitrite oxidation is also conserved in both genomes. Although both lack acidic proteomes synonymous with extreme halophiles, the pangenome of Ca. Nitromaritima RS specifically encodes enzymes with osmoregulatory and thermoprotective roles (i.e., ectoine/hydroxyectoine biosynthesis) and of thermodynamic importance (i.e., nitrate and nitrite reductases). Ca. Nitromaritima RS also possesses many hallmark traits of microaerophiles and high-affinity NOB. The abundance of the uncultured Ca. Nitromaritima lineage in marine oxyclines suggests their unrecognized ecological significance in deoxygenated areas of the global ocean.


Rubella metapopulation dynamics and importance of spatial coupling to the risk of congenital rubella syndrome in Peru.
Rubella is generally a mild childhood disease, but infection during early pregnancy may cause spontaneous abortion or congenital rubella syndrome (CRS), which may entail a variety of birth defects. Consequently, understanding the age-structured dynamics of this infection has considerable public health value. Vaccination short of the threshold for local elimination of transmission will increase the average age of infection. Accordingly, the classic concern for this infection is the potential for vaccination to increase incidence in individuals of childbearing age. A neglected aspect of rubella dynamics is how age incidence patterns may be moulded by the spatial dynamics inherent to epidemic metapopulations. Here, we use a uniquely detailed dataset from Peru to explore the implications of this for the burden of CRS. Our results show that the risk of CRS may be particularly severe in small remote regions, a prediction at odds with expectations in the endemic situation, and with implications for the outcome of vaccination. This outcome results directly from the metapopulation context: specifically, extinction-re-colonization dynamics are crucial because they allow for significant leakage of susceptible individuals into the older age classes during inter-epidemic periods with the potential to increase CRS risk by as much as fivefold.


A nemertean excitatory peptide/CCHamide regulates ciliary swimming in the larvae of Lineus longissimus.
The trochozoan excitatory peptide (EP) and its ortholog, the arthropod CCHamide, are neuropeptides that are only investigated in very few animal species. Previous studies on different trochozoan species focused on their physiological effect in adult specimens, demonstrating a myo-excitatory effect, often on tissues of the digestive system. The function of EP in the planktonic larvae of trochozoans has not yet been studied. We surveyed transcriptomes from species of various spiralian (Orthonectida, Nemertea, Brachiopoda, Entoprocta, Rotifera) and ecdysozoan taxa (Tardigrada, Onychophora, Priapulida, Loricifera, Nematomorpha) to investigate the evolution of EPs/CCHamides in protostomes. We found that the EPs of several pilidiophoran nemerteans show a characteristic difference in their C-terminus. Deorphanization of a pilidiophoran EP receptor showed, that the two splice variants of the nemertean Lineus longissimus EP activate a single receptor. We investigated the expression of EP in L. longissimus larvae and juveniles with customized antibodies and found that EP positive nerves in larvae project from the apical organ to the ciliary band and that EP is expressed more broadly in juveniles in the neuropil and the prominent longitudinal nerve cords. While exposing juvenile L. longissimus specimens to synthetic excitatory peptides did not show any obvious effect, exposure of larvae to either of the two EPs increased the beat frequency of their locomotory cilia and shifted their vertical swimming distribution in a water column upwards. Our results show that EP/CCHamide peptides are broadly conserved in protostomes. We show that the EP increases the ciliary beat frequency of L. longissimus larvae, which shifts their vertical distribution in a water column upwards. Endogenous EP may be released at the ciliary band from the projections of apical organ EP positive neurons to regulate ciliary beating. This locomotory function of EP in L. longissimus larvae stands in contrast to the repeated association of EP/CCHamides with its myo-excitatory effect in adult trochozoans and the general association with the digestive system in many protostomes.


The dentist's exposure to elemental mercury vapor during clinical work with amalgam.
Continuous measurements of mercury vapor in the breathing zone of the dentist were made under ordinary clinical conditions. Fifty old amalgam fillings were removed and replaced by new ones. The mercury vapor measurements were performed by means of atomic absorption spectrophotometry. On the basis of the type of suction device used, the measurements were divided into seven groups, each of which consisted of a series of measurements. During the cutting, filling, and polishing operations the mean mercury vapor levels in the breathing zone of the dentist were in the range of 1-2 micrograms Hg m-3 air when proper mercury hygiene measures were taken. This is far below the currently valid threshold limit value (30 micrograms Hg m-3 air) in Sweden. The saliva extractor and the dental mirror-evacuator did not influence the mercury vapor levels when used together with the high-volume evacuator. However, when only a saliva extractor was used, the cutting of amalgam fillings caused highly fluctuating mercury vapor levels, which were 2-15 times higher than the threshold limit value.


Sensory weighting and realignment: independent compensatory processes.
When estimating the position of one hand for the purpose of reaching to it with the other, humans have visual and proprioceptive estimates of the target hand's position. These are thought to be weighted and combined to form an integrated estimate in such a way that variance is minimized. If visual and proprioceptive estimates are in disagreement, it may be advantageous for the nervous system to bring them back into register by spatially realigning one or both. It is possible that realignment is determined by weights, in which case the lower-weighted modality should always realign more than the higher-weighted modality. An alternative possibility is that realignment and weighting processes are controlled independently, and either can be used to compensate for a sensory misalignment. Here, we imposed a misalignment between visual and proprioceptive estimates of target hand position in a reaching task designed to allow simultaneous, independent measurement of weights and realignment. In experiment 1, we used endpoint visual feedback to create a situation where task success could theoretically be achieved with either a weighting or realignment strategy, but vision had to be regarded as the correctly aligned modality to achieve success. In experiment 2, no endpoint visual feedback was given. We found that realignment operates independently of weights in the former case but not in the latter case, suggesting that while weighting and realignment may operate in conjunction in some circumstances, they are biologically independent processes that give humans behavioral flexibility in compensating for sensory perturbations.


Children as hematopoietic stem cell donors.
In the past half-century, hematopoietic stem cell transplantation has become standard treatment for a variety of diseases in children and adults, including selected hematologic malignancies, immunodeficiencies, hemoglobinopathies, bone marrow failure syndromes, and congenital metabolic disorders. There are 3 sources of allogeneic hematopoietic stem cells: bone marrow, peripheral blood, and umbilical cord blood; each has its own benefits and risks. Children often serve as hematopoietic stem cell donors, most commonly for their siblings. HLA-matched biological siblings are generally preferred as donors because of reduced risks of transplant-related complications as compared with unrelated donors. This statement includes a discussion of the ethical considerations regarding minors serving as stem cell donors, using the traditional benefit/burden calculation from the perspectives of both the donor and the recipient. The statement also includes an examination of the circumstances under which a minor may ethically participate as a hematopoietic stem cell donor, how the risks can be minimized, what the informed-consent process should entail, the role for a donor advocate (or some similar mechanism), and other ethical concerns. The American Academy of Pediatrics holds that minors can ethically serve as stem cell donors when specific criteria are fulfilled.


Palliative home care: health-related quality of life.
To determine the relationship between physical symptoms and health-related quality of life (HRQOL) of people enrolled in a palliative home care programme in Bogota, Colombia. A quantitative, observational, analytical, cross-sectional study. 123 people, chronically ill and enrolled in a palliative home care programme, participated. Convenience sampling was used, and data were collected during home visits. The HRQOL was perceived as fair by 42.3% of the participants, and the overall score was below the expected mean value (140.5). The psychological dimension score was also below the mean, while above-average results were obtained in the physical, social and spiritual dimensions. Pain (78.2%), insomnia (82.3%) and tiredness (77.4%) were the most altered symptoms. Statistically significant differences were found between HRQOL and educational level, occupation, low socioeconomic strata and religious commitment. Statistically significant associations were found between depression and the psychological dimension (r=0.208; p=0.020) and between depression and overall HRQOL (r=0.181; p=0.045). No relationship was observed between HRQOL and physical symptoms. Palliative home care programmes should thoroughly address psychological aspects, such as depression and anxiety, to ensure the best quality of life possible for the patients they assist.


Optical profiling of autonomous Ca2+ nanodomains generated by lysosomal TPC2 and TRPML1.
Multiple families of Ca2+-permeable channels co-exist on lysosomal Ca2+ stores but how each family couples to its own unique downstream physiology is unclear. We have therefore investigated the Ca2+-signalling architecture underpinning different channels on the same vesicle that drive separate pathways, using phagocytosis as a physiological stimulus. Lysosomal Ca2+-channels are a major Ca2+ source driving particle uptake in macrophages, but different channels drive different aspects of Fc-receptor-mediated phagocytosis: TPC2 couples to dynamin activation, whilst TRPML1 couples to lysosomal exocytosis. We hypothesised that they are driven by discrete local plumes of Ca2+ around open channels (Ca2+ nanodomains). To test this, we optimized Ca2+-nanodomain recordings by screening panels of genetically encoded Ca2+ indicators (GECIs) fused to TPC2 to monitor the [Ca2+] next to the channel. Signal calibration accounting for the distance of the GECI from the channel mouth reveals that, during phagocytosis, TPC2 generates local Ca2+ nanodomains around itself of up to 42 µM, nearly a hundred-fold greater than the global cytosolic [Ca2+] rise. We further show that TPC2 and TRPML1, though on the same lysosomes, generate autonomous Ca2+ nanodomains of high [Ca2+] that are largely insulated from one another, a platform allowing their discrete Ca2+-decoding to promote unique respective physiologies.


Infection-induced lung injury is worsened after renal buffering of hypercapnic acidosis.
Prolonged hypercapnia is commonly encountered during the treatment of acute respiratory distress syndrome and acute respiratory failure attributable to other causes with protective ventilation strategies. In these circumstances, compensatory renal buffering returns pH to normal establishing a condition of buffered hypercapnia. It is also common intensive care practice to correct the pH more rapidly using bicarbonate infusions. Although it is well-established that hypercapnic acidosis has potent anti-inflammatory and protective effects, the effect of buffered hypercapnia on acute lung injury and acute respiratory distress syndrome is unknown. We therefore wished to determine the effects of buffered hypercapnia on acute lung injury induced by endotoxin or Escherichia coli infection in vivo. Prospective, randomized animal study. University research laboratory. Adult male Sprague-Dawley rats. We established buffered hypercapnia by exposing rats to a hypercapnic environment for 3 days before the induction of lung injury. Buffered hypercapnia rats (initial pH >7.35, FiCO2 = 0.05) and normocapnic controls (initial pH >7.35, FiCO2 = 0.00) were then anesthetized, mechanically ventilated, and lung injury induced by intra-tracheal inoculation of endotoxin (series I) or Escherichia coli (series II). Buffered hypercapnia significantly increased both endotoxin and Escherichia coli-induced lung injury when compared to normocapnic controls, as assessed by arterial oxygenation, lung compliance, pro-inflammatory pulmonary cytokine concentrations, and measurements of structural lung damage. In additional in vitro experiments buffered hypercapnia did not alter neutrophil phagocytosis ability but did impaired epithelial wound healing. Our results demonstrate that infection-induced injury in vivo is worsened after renal buffering of hypercapnic acidosis independently of any changes in tidal volume. These findings have important implications for our understanding of the pathogenesis of infection-induced lung injury during the use protective ventilation strategies that permits buffered hypercapnia and during infective exacerbations of chronic lung diseases associated with sustained hypercapnia.


Human papillomavirus in transitional cell carcinoma of the urinary bladder
The presence of human papillomavirus (HPV) was detected in transitional cell carcinoma of the urinary bladder. PCR amplification of DNA from 71 tumors, using consensus primers for a fragment of the L1 gene, detected 6 strongly positive tumors (through ethidium bromide staining of a gel) and 22 moderately positive tumors (through Southern blotting of the amplified DNA) for a total of 28 (39%) of the tumors. The presence of HPV was correlated with grade but not stage of the tumors. Typing of HPV was performed on 31 tumors: all positive tumors contained HPV 16 DNA except for one Ta tumor which contained HPV 11 DNA. Our data also showed a large variability in the sensitivity of HPV DNA detection, depending on sample fixation, DNA preparation, and amplification conditions, which may explain in large part the discrepancies reported in the literature on the association of HPV to bladder cancer. Because of the low HPV DNA copy number observed in bladder tumors, our results suggest that HPV should ideally be tested on fresh or frozen tumor material, that SDS detergent should be avoided for the preparation of DNA, and that the amplification conditions are critical for optimal detection.


Radiation dose-dependent changes in tumor metabolism measured by 31P nuclear magnetic resonance spectroscopy.
The effects of radiation dose upon a hypoxic murine mammary carcinoma were followed using 31P nuclear magnetic resonance spectroscopy. Animals were studied before and over the course of 9 days after tumors were irradiated with a single dose of 0, 4, 8, or 17 Gy. The current data is compared to our previous studies of the effects of 32 or 65 Gy on the same tumor model. The energy status of the tumors, as reflected in nucleotide triphosphate:Pi and phosphocreatine:Pi ratios, improved after receiving a dose of 8 to 65 Gy and decreased after receiving 0 or 4 Gy doses. The energy status of the 8- to 65-Gy dose cohorts reached a maximum between 1 and 4 days after irradiation. Additionally, the change in the hypoxic cell fraction 48 h after a 17-Gy dose was determined; it was calculated from changes in the doses required to control 50% of the tumors post radiation for clamped (hypoxic) and unclamped (normoxic) tumors in parallel animal cohorts. A significant decrease compared to preirradiation values was observed in the hypoxic cell fraction following 17 Gy irradiation. This decrease was temporally coincident with increases in tumor energy status measured using nuclear magnetic resonance and was similar to our previously reported results of the change in hypoxic fraction 48 h after a 32-Gy dose. Changes in the relative ratio of phosphomonoesters showed a strong dose dependence after irradiation. The downfield component of the phosphomonoester peak, which consists largely of phosphoethanolamine, increased relative to the upfield component, phosphocholine. This dose-dependent ratio reached a maximum approximately 7 days post radiation. Changes in the levels of membrane phospholipid precursors may be related to alterations in cell proliferation or may be a result of radiation-induced membrane damage.


The Design of SimpleITK.
SimpleITK is a new interface to the Insight Segmentation and Registration Toolkit (ITK) designed to facilitate rapid prototyping, education and scientific activities via high level programming languages. ITK is a templated C++ library of image processing algorithms and frameworks for biomedical and other applications, and it was designed to be generic, flexible and extensible. Initially, ITK provided a direct wrapping interface to languages such as Python and Tcl through the WrapITK system. Unlike WrapITK, which exposed ITK's complex templated interface, SimpleITK was designed to provide an easy to use and simplified interface to ITK's algorithms. It includes procedural methods, hides ITK's demand driven pipeline, and provides a template-less layer. Also SimpleITK provides practical conveniences such as binary distribution packages and overloaded operators. Our user-friendly design goals dictated a departure from the direct interface wrapping approach of WrapITK, toward a new facade class structure that only exposes the required functionality, hiding ITK's extensive template use. Internally SimpleITK utilizes a manual description of each filter with code-generation and advanced C++ meta-programming to provide the higher-level interface, bringing the capabilities of ITK to a wider audience. SimpleITK is licensed as open source software library under the Apache License Version 2.0 and more information about downloading it can be found at http://www.simpleitk.org.


Recombinant DNA technology for melanoma immunotherapy: anti-Id DNA vaccines targeting high molecular weight melanoma-associated antigen.
Anti-idiotypic MK2-23 monoclonal antibody (anti-Id MK2-23 mAb), which mimics the high molecular weight melanoma-associated antigen (HMW-MAA), has been used to implement active immunotherapy against melanoma. However, due to safety and standardization issues, this approach never entered extensive clinical trials. In the present study, we investigated the usage of DNA vaccines as an alternative to MK2-23 mAb immunization. MK2-23 DNA plasmids coding for single chain (scFv) MK2-23 antibody were constructed via the insertion of variable heavy (V H) and light (V L) chains of MK2-23 into the pVAC-1mcs plasmids. Two alternative MK2-23 plasmids format V H/V L, and V L/V H were assembled. We demonstrate that both polypeptides expressed by scFv plasmids in vitro retained the ability to mimic HMW-MAA antigen, and to elicit specific anti-HMW-MAA humoral and cellular immunoresponses in immunized mice. Notably, MK2-23 scFv DNA vaccines impaired the onset and growth of transplantable B16 melanoma cells not engineered to express HMW-MAA. This pilot study suggests that optimized MK2-23 scFv DNA vaccines could potentially provide a safer and cost-effective alternative to anti-Id antibody immunization, for melanoma immunotherapy.


Regulatable systemic production of monoclonal antibodies by in vivo muscle electroporation.
The clinical application of monoclonal antibodies (mAbs) potentially concerns a wide range of diseases including, among others, viral infections, cancer and autoimmune diseases. Although intravenous infusion appears to be the simplest and most obvious mode of administration, it is very often not applicable to long-term treatments because of the restrictive cost of mAbs certified for human use and the side effects associated with injection of massive doses of antibodies. Gene/cell therapies designed for sustained and, possibly, regulatable in vivo production and systemic delivery of mAbs might permit to advantageously replace it. We have already shown that several such approaches allow month- to year-long ectopic antibody production by non-B cells in living organisms. Those include grafting of ex vivo genetically modified cells of various types, in vivo adenoviral gene transfer and implantation of encapsulated antibody-producing cells. Because intramuscular electrotransfer of naked DNA has already been used for in vivo production of a variety of proteins, we have wanted to test whether it could be adapted to that of ectopic mAbs as well. We report here that this is actually the case since both long-term and regulatable production of an ectopic mAb could be obtained in the mouse taken as a model animal. Although serum antibody concentrations obtained were relatively low, these data are encouraging in the perspective of future therapeutical applications of this technology in mAb-based immunotherapies, especially in developing countries where cost-effective and easily implementable technologies would be required for large-scale applications in the context of severe chronic viral diseases such as HIV and HCV infections.


Evolution of primate ABO blood group genes and their homologous genes.
There are three common alleles (A, B, and O) at the human ABO blood group locus. We compared nucleotide sequences of these alleles, and relatively large numbers of nucleotide differences were found among them. These differences correspond to the divergence time of at least a few million years, which is unusually large for a human allelic divergence under neutral evolution. We constructed phylogenetic networks of human and nonhuman primate ABO alleles, and at least three independent appearances of B alleles from the ancestral A form were observed. These results suggest that some kind of balancing selection may have been operating at the ABO locus. We also constructed phylogenetic trees of ABO and their evolutionarily related alpha-1,3-galactosyltransferase genes, and the divergence time between these two families was estimated to be roughly 400 MYA.


Mutations in the microtubule-associated protein 1A (Map1a) gene cause Purkinje cell degeneration.
The structural microtubule-associated proteins (MAPs) are critical for the organization of neuronal microtubules (MTs). Microtubule-associated protein 1A (MAP1A) is one of the most abundantly expressed MAPs in the mammalian brain. However, its in vivo function remains largely unknown. Here we describe a spontaneous mouse mutation, nm2719, which causes tremors, ataxia, and loss of cerebellar Purkinje neurons in aged homozygous mice. The nm2719 mutation disrupts the Map1a gene. We show that targeted deletion of mouse Map1a gene leads to similar neurodegenerative defects. Before neuron death, Map1a mutant Purkinje cells exhibited abnormal focal swellings of dendritic shafts and disruptions in axon initial segment (AIS) morphology. Furthermore, the MT network was reduced in the somatodendritic and AIS compartments, and both the heavy and light chains of MAP1B, another brain-enriched MAP, was aberrantly distributed in the soma and dendrites of mutant Purkinje cells. MAP1A has been reported to bind to the membrane-associated guanylate kinase (MAGUK) scaffolding proteins, as well as to MTs. Indeed, PSD-93, the MAGUK specifically enriched in Purkinje cells, was reduced in Map1a(-/-) Purkinje cells. These results demonstrate that MAP1A functions to maintain both the neuronal MT network and the level of PSD-93 in neurons of the mammalian brain.


How safe are children with COVID-19 from cardiac risks? Pediatric risk assesment; insights from echocardiography and electrocardiography
Approximately 40 million individuals worldwide have been infected with SARS-CoV-2, the virus responsible for the novel coronavirus disease-2019 (COVID-19). Despite the current literature about the cardiac effects of COVID-19 in children, more information is required. We aimed to determine both cardiovascular and arrhythmia assessment via electrocardiographic and echocardiographic parameters. We evaluated seventy children who were hospitalized with COVID-19 infections and seventy children as normal control group through laboratory findings, electrocardiography (ECG), and transthoracic echocardiography (TTE). We observed significantly increased levels of Tp-Te, Tp-Te/QT, and Tp-Te/QTc compared with the control group. Twenty-five of 70 (35.7%) patients had fragmented QRS (fQRS) without increased troponin levels. On the other hand, none of the patients had pathologic corrected QT(QTc) prolongation during the illness or its treatment. On TTE, 20 patients had mild mitral insufficiency, among whom only five had systolic dysfunction (ejection fraction < 55%). There was no significant difference between the patient and control groups, except for isovolumic relaxation time (IVRT) in terms of mean systolic and diastolic function parameters. IVRT of COVID patients was significantly lower than that of control group. Despite all the adult studies, the effects of COVID‐19 on myocardial function are not well established in children. The thought that children are less affected by the illness may be a misconception.


Thiamine Deficiency in Tropical Pediatrics: New Insights into a Neglected but Vital Metabolic Challenge.
In humans, thiamine is a micronutrient prone to depletion that may result in severe clinical abnormalities. This narrative review summarizes current knowledge on thiamine deficiency (TD) and bridges the gap between pathophysiology and clinical presentation by integrating thiamine metabolism at subcellular level with its function to vital organs. The broad clinical spectrum of TD is outlined, with emphasis on conditions encountered in tropical pediatric practice. In particular, TD is associated with type B lactic acidosis and classic forms of beriberi in children, but it is often unrecognized. Other severe acute conditions are associated with hypermetabolism, inducing a functional TD. The crucial role of thiamine in infant cognitive development is also highlighted in this review, along with analysis of the potential impact of TD in refeeding syndrome during severe acute malnutrition (SAM). This review aims to increase clinical awareness of TD in tropical settings where access to diagnostic tests is poor, and advocates for an early therapeutic thiamine challenge in resource-limited settings. Moreover, it provides evidence for thiamine as treatment in critical conditions requiring metabolic resuscitation, and gives rationale to the consideration of increased thiamine supplementation in therapeutic foods for malnourished children.


Redefining fine roots improves understanding of below-ground contributions to terrestrial biosphere processes.
Fine roots acquire essential soil resources and mediate biogeochemical cycling in terrestrial ecosystems. Estimates of carbon and nutrient allocation to build and maintain these structures remain uncertain because of the challenges of consistently measuring and interpreting fine-root systems. Traditionally, fine roots have been defined as all roots ≤ 2 mm in diameter, yet it is now recognized that this approach fails to capture the diversity of form and function observed among fine-root orders. Here, we demonstrate how order-based and functional classification frameworks improve our understanding of dynamic root processes in ecosystems dominated by perennial plants. In these frameworks, fine roots are either separated into individual root orders or functionally defined into a shorter-lived absorptive pool and a longer-lived transport fine-root pool. Using these frameworks, we estimate that fine-root production and turnover represent 22% of terrestrial net primary production globally - a c. 30% reduction from previous estimates assuming a single fine-root pool. Future work developing tools to rapidly differentiate functional fine-root classes, explicit incorporation of mycorrhizal fungi into fine-root studies, and wider adoption of a two-pool approach to model fine roots provide opportunities to better understand below-ground processes in the terrestrial biosphere.


Assessing the performance of general practices caring for patients with asthma.
General practitioners (GPs) have had to record information about chronic disease management and send this to the health authorities since the introduction of the new contract in 1990. To discover the relationship between practice emergency admission rates for asthma and the characteristics of the practice's patients and chronic disease management programme. This was an ecological study comparing practice emergency admission rates of asthma by general practice with the practice's performance in measuring the prevalence, peak flow, and number of patients on regular prophylaxis. In addition, prescribing analysis and costing (PACT) data and census linkage were used to assign social class to patients and, when aggregated, to practices. The practice admission rate was correlated against each of these variables and then the relationships were explored in multiple linear regressions. A high rate of admission in practices was correlated with deprivation of the patients, in the form of a practice Townsend score (r = 0.33, P = 0.003), and also with poorer prescribing, measured by the preventer-reliever ratio (r = -0.38, P = 0.001). Regression analysis showed that the relationship between good prescribing and low admission rates was not explained by confounding variables. Only 32% of the variation in admission rates between practices was explained by the regression equation. None of the variables recorded in the annual report were significantly related to admission rates. Annual reports from the practice to the health authority are unhelpful in monitoring general practice performance, but prescribing, as measured by the preventer-reliever ratio, and hospital admission rates have limited usefulness.


Putting down the revolt: Enactivism as a philosophy of nature.
Enactivists frequently argue their account heralds a revolution in cognitive science: enactivism will unseat cognitivism as the dominant paradigm. We examine the lines of reasoning enactivists employ in stirring revolt, but show that none of these prove compelling reasons for cognitivism to be replaced by enactivism. First, we examine the hard sell of enactivism: enactivism reveals a critical explanatory gap at the heart of cognitivism. We show that enactivism does not meet the requirements to incite a paradigm shift in the Kuhnian sense-there is no internal crisis in cognitivism. Nor does it provide inherently better explanations of cognition as some have claimed. Second, we consider the soft sell of enactivism: enactivism provides a more attractive, parsimonious, or clear-eyed lens on cognition. This move proves to boil down to a misunderstanding of how theories are selected in science. Instead we lend support to a broader and more desirable way to conceive of enactivism, the recent proposal that enactivism is a philosophy of nature. We explain how a philosophy of nature does more than support a single research paradigm by integrating scientific questions into a cohesive picture.


Diels-Alder reaction mechanisms of substituted chiral anthracene: A theoretical study based on the reaction force and reaction electronic flux.
Quantum chemical calculations were used to study the mechanism of Diels-Alder reactions involving chiral anthracenes as dienes and a series of dienophiles. The reaction force analysis was employed to obtain a detailed scrutiny of the reaction mechanisms, it has been found that thermodynamics and kinetics of the reactions are quite consistent: the lower the activation energy, the lower the reaction energy, thus following the Bell-Evans-Polanyi principle. It has been found that activation energies are mostly due to structural rearrangements that in most cases represented more than 70% of the activation energy. Electronic activity mostly due to changes in σ and π bonding were revealed by the reaction electronic flux (REF), this property helps identify whether changes on σ or π bonding drive the reaction. Additionally, new global indexes describing the behavior of the electronic activity were introduced and then used to classify the reactions in terms of the spontaneity of their electronic activity. Local natural bond order electronic population analysis was used to check consistency with global REF through the characterization of specific changes in the electronic density that might be responsible for the activity already detected by the REF. Results show that reactions involving acetoxy lactones are driven by spontaneous electronic activity coming from bond forming/strengthening processes; in the case of maleic anhydrides and maleimides it appears that both spontaneous and non-spontaneous electronic activity are quite active in driving the reactions.


CD22, a B lymphocyte-specific adhesion molecule that regulates antigen receptor signaling.
The development of B lymphocytes is a highly regulated process that depends in part on lineage-specific cell surface molecules. In addition, transmembrane signals generated through the B cell antigen receptor and other surface molecules regulate B cell responses to foreign antigens. Recent studies reveal CD22 to be a functionally significant receptor during these processes. CD22 is first expressed in the cytoplasm of pro-B and pre-B cells, and on the surface as B cells mature to become IgD+. CD22 is a member of the Ig superfamily that serves as an adhesion receptor for sialic acid-bearing ligands expressed on erythrocytes and all leukocyte classes. In addition to its potential role as a mediator of intercellular interactions, signal transduction through CD22 can activate B cells and modulate antigen receptor signaling in vitro. CD22 signaling is mediated via interactions with a number of kinases and phosphatases that bind the cytoplasmic domain through phosphorylated tyrosine residues located within consensus TAM and TIM motifs. The phenotype of CD22-deficient mice suggests that CD22 is primarily involved in the generation of mature B cells within the bone marrow, blood, and marginal zones of lymphoid tissues. Most notable in CD22-deficient mice is a significant diminution of surface Ig levels in these B cell subpopulations, which suggests that CD22 functions in vivo to adjust the signaling threshold of cell surface antigen receptors. A further understanding of CD22 function is required and may reveal roles for CD22 in disease susceptibility or the development of autoimmunity.


Molecular characterisation of two structurally distinct groups of human homers, generated by extensive alternative splicing.
Homer proteins bind specifically to the C termini of the metabotropic glutamate receptor mGluR1alpha/a and mGluR5, play a role in their targeting and modulate their synaptic properties. We have discovered that extensive alternative splicing generates a family of 17 Homer proteins. These fall into two distinct groups of 12 "long" Homers, which all have a coiled-coil domain at their C termini, and five "short" Homers, which lack such a domain. All Homers contain the N-terminal sequence responsible for their binding to mGluR1alpha/a receptors and can be co-localised with the recombinantly expressed mGluR1alpha/a protein in HEK-293 cells. The existence of the long and the short variants of each of the Homer-1, Homer-2 and Homer-3 proteins reflects the fundamental principles of Homer functions.


Profiling of bacterial flora in crevices around titanium orthodontic anchor plates.
The aims of this study were to characterize the microflora in crevices around titanium orthodontic anchor plates using anaerobic culture and molecular biological techniques for bacterial identification, and to compare the microbial composition between crevices around anchor plates and gingival crevices. Samples from crevices around titanium anchor plates and healthy gingival crevices of 17 subjects (aged 20-29) were cultured anaerobically, and isolated bacteria were identified by 16S rRNA sequencing. The average logarithm colony-forming units/ml were 6.84, 7.51 and 8.88 in healthy anchor plate crevices, inflamed anchor plate crevices and healthy gingival crevices, respectively, indicating that the bacterial density of anchor plate crevices was lower than that of healthy gingival crevices. Of 184 strains isolated from healthy anchor plate crevices of seven subjects, 108 (59%) were anaerobic bacteria, while 73 (40%) were facultative bacteria. Predominant isolates were Gram-negative rods, such as Campylobacter (12%), Fusobacterium (10%) and Selenomonas (10%), and Gram-positive facultative bacteria, such as Actinomyces (17%) and Streptococcus (8.2%). Of 133 strains isolated from inflamed anchor plate crevices of three subjects, 110 (83%) were anaerobic bacteria, while predominant isolates were Gram-negative rods, such as Prevotella (47%), Fusobacterium (33%) and Campylobacter (16%). On the other hand, of 146 strains isolated from healthy gingival crevices of seven subjects, 98 (67%) were facultative bacteria, while 45 (31%) were anaerobic bacteria. Predominant isolates were Gram-positive facultative bacteria, such as Actinomyces (37%) and Streptococcus (20%). These results suggest that the environment in crevices around titanium orthodontic anchor plates is anaerobic and supportive of anaerobic growth of bacteria, which may trigger inflammation in the tissue around the plates.


Progress in the synthesis and exploitation of catenanes since the Millennium.
Catenanes - molecules consisting of interlocked macrocyclic rings - have been prepared by templation strategies for some thirty years. The utilization of Cu(I) cation, aromatic donor-acceptor interactions and hydrogen bonding assisted self-assembly strategies has led to the construction of numerous examples of these aesthetically pleasing species. This review seeks to discuss key developments in the synthesis and functional application of catenanes that have occurred since the Millennium. The much expanded range of metal cation templates; the genesis and growth of anion templation, as well as the use of alternative supramolecular interactions (halogen bonding and radical templation) and thermodynamically controlled reactions to synthesize catenanes are detailed. The class of catenanes that may be described as "molecular machines" are then highlighted and to conclude, attempts to fabricate catenanes onto surfaces and into metal organic frameworks (MOFs) are discussed.


Paclitaxel combined with siRNA targeting HPV16 oncogenes improves cytotoxicity for cervical carcinoma.
Cervical cancer is attributable to continuous expression of the E6 and E7 oncoproteins of the high-risk human papillomaviruses. These proteins target p53 and members of the retinoblastoma cellular regulatory protein family respectively for degradation, disrupting cellular control over apoptosis, senescence and the cell cycle. Delivery of short interfering RNAs (siRNAs) targeting mRNA from the HPV16 E6/E7 open reading frame to HPV16-positive cell lines, led to an 80% reduction in full-length transcripts and 60% reduction in total (full-length and spliced) transcripts. Downregulation of E6 mRNA led to increased levels of p53 detectable by western blot and resulted in an eightfold increase in luciferase expression from a p53-responsive reporter plasmid. Downregulation of E7 mRNA reduced the levels of E7 protein and increased the levels of hypophosphorylated pRb. Cellular proliferation was reduced after siRNA delivery; the effect being greater in SiHa cells than CaSki cells and when full-length transcripts encoding E6 and spliced transcripts encoding E7 were both targeted. There was no loss of proliferation in human papillomavirus-negative cell lines. Elevation of p53 in the absence of changes to Rb led to 35% of CaSki cells undergoing apoptosis, whereas in SiHa cells restoration of both p53 and hypophosphorylated Rb had the greatest effect with 25% of cells undergoing apoptosis. The combined use of oncogene-targeting siRNA with either carboplatin, irinotecan, leptomycin B or doxorubicin led to additive toxicity, whereas, with cisplatin, led to sub-additive toxicity. In contrast, siRNA combined with paclitaxel treatment resulted in synergistic toxicity, with intronic siRNA (which mainly targets E6) more effective than exonic siRNA (which targets both E6 and E7). The growth of SiHa xenograft tumors was reduced using paclitaxel combined with intronic and exonic siRNA, compared with exonic siRNA alone, confirming the synergistic relationship between p53 restoration and paclitaxel.


Interactive entity resolution in relational data: a visual analytic tool and its evaluation.
Databases often contain uncertain and imprecise references to real-world entities. Entity resolution, the process of reconciling multiple references to underlying real-world entities, is an important data cleaning process required before accurate visualization or analysis of the data is possible. In many cases, in addition to noisy data describing entities, there is data describing the relationships among the entities. This relational data is important during the entity resolution process; it is useful both for the algorithms which determine likely database references to be resolved and for visual analytic tools which support the entity resolution process. In this paper, we introduce a novel user interface, D-Dupe, for interactive entity resolution in relational data. D-Dupe effectively combines relational entity resolution algorithms with a novel network visualization that enables users to make use of an entity's relational context for making resolution decisions. Since resolution decisions often are interdependent, D-Dupe facilitates understanding this complex process through animations which highlight combined inferences and a history mechanism which allows users to inspect chains of resolution decisions. An empirical study with 12 users confirmed the benefits of the relational context visualization on the performance of entity resolution tasks in relational data in terms of time as well as users' confidence and satisfaction.


A novel postsynaptic mechanism for heterosynaptic sharing of short-term plasticity.
Postsynaptic release of Ca(2+) from intracellular stores is an important means of cellular signaling that mediates numerous forms of synaptic plasticity. Previous studies have identified a postsynaptic intracellular Ca(2+) requirement for a form of short-term plasticity, post-tetanic potentiation (PTP) at sensory neuron (SN)-motor neuron synapses in Aplysia. Here, we show that postsynaptic IP(3)-mediated Ca(2+) release in response to a presynaptic tetanus in an SN that induces PTP can confer transient plasticity onto a neighboring SN synapse receiving subthreshold activation. This heterosynaptic sharing of plasticity represents a dynamic, short-term synaptic enhancement of synaptic inputs onto a common postsynaptic target. Heterosynaptic sharing is blocked by postsynaptic disruption of Ca(2+)- and IP(3)-mediated signaling, and, conversely, it is mimicked by postsynaptic injection of nonhydrolyzable IP(3), and by photolysis of caged IP(3) in the MN. The molecular mechanism for heterosynaptic sharing involves metabotropic glutamate receptors and Homer-dependent interactions, indicating that Homer can facilitate the integration of Ca(2+)-dependent plasticity at neighboring postsynaptic sites and provides a postsynaptic mechanism for the spread of plasticity induced by presynaptic activation. Our results support a model in which postsynaptic summation of IP(3) signals from suprathreshold and subthreshold inputs results in molecular coincidence detection that gives rise to a novel form of heterosynaptic plasticity.


Clinical, imaging and genotypical features of three deceased and five surviving cases with ADA2 deficiency.
Deficiency of adenosine deaminase type 2 (DADA2) is a rare form of autoinflammatory disorder with limited reported cases. In this paper, we have presented the clinico-immunological, radiological and genetic characteristics of five surviving and three deceased childhood-onset DADA2 patients. We aimed to compare surviving and deceased patients in terms of clinical features and treatment modalities. Moreover, we have evaluated the causes of death in our DADA2 subjects together with the previously reported cases. Demographic features, clinical characteristics, imaging findings, mutations and pharmacological treatments of DADA2 subjects were noted from patient records of pediatric and adult rheumatology clinics in a retrospective and longitudinal nature. Eight patients from seven families were enrolled. While five of them were surviving, three of them had died due to various reasons. Median age of the patients at disease onset and diagnosis was 7 years (range 0.5-13 years) and 14 years (range 5-27 years), respectively. The main clinical manifestations were cutaneous findings (7/8), recurrent low-grade fever (6/8), neurological involvement (6/8) and gastrointestinal involvement (5/8). All patients had increased acute phase reactants at presentation and also during the disease flares. Until the diagnosis of DADA2 was confirmed, five patients have been followed-up with the diagnosis of PAN: two patients both with PAN and FMF, and one patient with CAPS and vasculitis. Demographic, clinical, neurological features and genetic mutations did not differ in surviving and deceased DADA2 patients. Deceased and surviving subjects differed in terms of treatment modalities after the diagnosis of DADA2. Anti-TNF alpha treatment has been initiated in five surviving patients as soon as the diagnosis of DADA2 was established. However, three patients who have died were not able to use sufficient doses of anti-TNF alpha treatment; in one case due to reluctance of patient and in two cases due to establishment of the definite diagnosis by genetic analysis at the same time with the last fatal DADA2 episode. Despite limited number of patients, this case series for the first time compares the phenotypic, genotypic and medication differences between surviving and deceased DADA2 patients. Anti-TNF alpha treatment seems to be efficient and lifesaving in DADA2 patients.


Effortful pitch glide: a potential new exercise evaluated by dynamic MRI.
The purpose of this study was to compare the biomechanics of the effortful pitch glide (EPG) with swallowing using dynamic MRI. The EPG is a combination of a pitch glide and a pharyngeal squeeze maneuver for targeting laryngeal and pharyngeal muscles. The authors hypothesized that the EPG would elicit significantly greater structural excursions of anterior hyoid, superior hyoid, hyolaryngeal approximation, laryngeal elevation, and lateral pharyngeal wall medialization compared with swallowing. Eleven healthy, young subjects with a mean age of 25 were recruited. The EPG was first taught and verified via laryngoscopy. Then 2-planar (coronal and sagittal) dynamic MRI acquisitions captured 10 repeated swallows and 3 EPGs. Kinematic analyses of minimum and maximum excursion of anatomical landmarks were calculated. Results showed a nonsignificant difference between the 2 tasks for range of excursion with all measured biomechanics except for superior hyoid, where the swallow showed significantly greater excursion. This indicated that swallowing and EPG biomechanics were comparable, lending support for the potential use of the EPG as another nonswallowing exercise. Findings suggest EPG may be an effective exercise to target several important swallowing muscles, especially the long pharyngeal muscles that elevate the larynx and shorten the pharynx in swallowing.


Is it Time to Update How Suspected Angina Is Evaluated prior to the Use of Specialized Tests Implications Based on a Systematic Review.
Appropriate use of specialized tests to assess chest pain is based classically on minimal information such as age, gender and the patient's description of pain. This approach has not been reevaluated in decades. We examined the relationship between history, examination and routine laboratory tests to identify factors warranting prospective validation as predictors of underlying coronary artery disease (CAD). Studies linking obstructive CAD (≥50% diameter stenosis of at least one vessel by invasive angiography or cardiac computed tomographic angiography) and elements of history, examination and laboratory tests were identified. Forty-one prospectively identified papers were analyzed. Advanced age, gender and chest pain descriptors were extremely important, although the last was less so in women, in whom the presence of risk factors may be more important. Physical examination and chest X-ray were largely noncontributory. Laboratory tests were of variable utility other than to identify risk factors not already known from the history. However, biomarkers such as troponin, brain natriuretic factor and inflammatory markers were promising. The electrocardiogram was mainly important for the identification of ST-T abnormalities. This review identifies the most promising factors warranting prospective validation for improving the pretest probability estimation of CAD, so appropriate use criteria for the utilization of specialized diagnostic tests can be updated and improved.


Adaptation of the Freshwater Bloom-Forming Cyanobacterium Microcystis aeruginosa to Brackish Water Is Driven by Recent Horizontal Transfer of Sucrose Genes.
Microcystis aeruginosa is a bloom-forming cyanobacterium found in eutrophic water bodies worldwide. M. aeruginosa blooms usually occur in freshwater; however, they have also been reported to occur in brackish water. Because M. aeruginosa often produces the cyanotoxin microcystin, they are a major concern to public health and environment. Despite this, the ecology, genomic basis, and evolutionary process underlying the M. aeruginosa bloom invasion from fresh to brackish water have been poorly investigated. Hence, in the present study, we have sequenced and characterized genomes of two newly discovered salt-tolerant M. aeruginosa strains obtained from Japanese brackish water lakes (Lakes Shinji and Tofutsu). Both genomes contain a set of genes for the synthesis of osmolyte sucrose (sppA, spsA, and susA), hitherto identified in only one strain (PCC 7806) of M. aeruginosa. Chemical and gene expression analyses confirmed sucrose accumulation induced by salt. A comprehensive genetic survey of >200 strains indicated that sucrose genes are extremely rare in M. aeruginosa. Most surprisingly, comparative genome analyses of the three strains indicated extremely low genetic diversity in the sucrose genes compared with other core genome genes, suggesting very recent acquisitions via horizontal transfer. Invasion of M. aeruginosa blooms into brackish water may be a recent event triggered by anthropogenic eutrophication of brackish water.


Visual temporal integration windows are adult-like in 5- to 7-year-old children.
The visual system must organize dynamic input into useful percepts across time, balancing between stability and sensitivity to change. The temporal integration window (TIW) has been hypothesized to underlie this balance: If two or more stimuli fall within the same TIW, they are integrated into a single percept; those that fall in different windows are segmented (Arnett & Di Lollo, 1979; Wutz, Muschter, van Koningsbruggen, Weisz, & Melcher, 2016). Visual TIWs have been studied in adults, showing average windows of 65 ms (Wutz et al., 2016); however, it is unclear how windows develop through early childhood. Here we measured TIWs in 5- to 7-year-old children and adults, using a variant of the missing dot task (Di Lollo, 1980; Wutz et al. 2016), in which integration and segmentation thresholds were measured within the same participant, using the same stimuli. Participants saw a sequence of two displays separated by an interstimulus interval (ISI) that determined the visibility of a visual search target. Longer ISIs increased the likelihood of detecting a segmentation target (but decreased detection for the integration target) although shorter ISIs increased the likelihood of detecting the integration target (but decreased detection of the segmentation target). We could then estimate the TIW by measuring the point at which these two functions intersect. Children's TIWs (M = 68 ms) were comparable to adults' (M = 73 ms) with no appreciable age trend within our sample, indicating that TIWs reach adult levels by approximately 5 years of age.


Left anterior small thoracotomy for minimally invasive coronary artery bypass grafting.
Since the 1990 s, minimally invasive cardiac surgery has gained wide acceptance due to patient and economic demand. The advantages are less trauma, bleeding, wound infections, pain and faster recovery. Many studies showed that the outcomes are comparable with those of conventional sternotomy. Left anterior small thoracotomy (LAST) evolved into a routine and safe access in specialized centres for minimally invasive direct coronary artery bypass grafting. The 6-cm incision is localized above the fourth intercostal space, 3-4 cm lateral to the left sternal border and below the left mammilla. With a double-lumen tube, the left lung is deflated before entering the pleural space. The left internal mammary artery is harvested under direct vision with the use of special retractors. The anastomosis of the left anterior descending artery is performed on the beating heart as known from off-pump surgery. One chest tube is inserted. The intercostal space is closed with braided sutures to prevent lung herniation. Ropivacaine is used for local infiltration. The pectoral muscle, subcutaneous tissue and skin are closed with running sutures. Complications of the LAST approach are rare (conversion to sternotomy, re-thoracotomy, phrenic nerve palsy, wound infection and thoracic wall hernia) and well manageable.


Investigation of specific substitutions in virulence genes characterizing phenotypic groups of low-virulence field strains of Listeria monocytogenes.
Several models have shown that virulence varies from one strain of Listeria monocytogenes to another, but little is known about the cause of low virulence. Twenty-six field L. monocytogenes strains were shown to be of low virulence in a plaque-forming assay and in a subcutaneous inoculation test in mice. Using the results of cell infection assays and phospholipase activities, the low-virulence strains were assigned to one of four groups by cluster analysis and then virulence-related genes were sequenced. Group I included 11 strains that did not enter cells and had no phospholipase activity. These strains exhibited a mutated PrfA; eight strains had a single amino acid substitution, PrfAK220T, and the other three had a truncated PrfA, PrfADelta174-237. These genetic modifications could explain the low virulence of group I strains, since mutated PrfA proteins were inactive. Group II and III strains entered cells but did not form plaques. Group II strains had low phosphatidylcholine phospholipase C activity, whereas group III strains had low phosphatidylinositol phospholipase C activity. Several substitutions were observed for five out of six group III strains in the plcA gene and for one out of three group II strains in the plcB gene. Group IV strains poorly colonized spleens of mice and were practically indistinguishable from fully virulent strains on the basis of the above-mentioned in vitro criteria. These results demonstrate a relationship between the phenotypic classification and the genotypic modifications for at least group I and III strains and suggest a common evolution of these strains within a group.


Coronavirus vaccine development: from SARS and MERS to COVID-19.
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a new type of coronavirus that causes the Coronavirus Disease 2019 (COVID-19), which has been the most challenging pandemic in this century. Considering its high mortality and rapid spread, an effective vaccine is urgently needed to control this pandemic. As a result, the academia, industry, and government sectors are working tightly together to develop and test a variety of vaccines at an unprecedented pace. In this review, we outline the essential coronavirus biological characteristics that are important for vaccine design. In addition, we summarize key takeaways from previous vaccination studies of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV), highlighting the pros and cons of each immunization strategy. Finally, based on these prior vaccination experiences, we discuss recent progress and potential challenges of COVID-19 vaccine development.


Association between brain temperature and dentate field potentials in exploring and swimming rats.
Attempts to correlate behavioral learning with cellular changes, such as increased synaptic efficacy, have often relied on increased extracellular potentials as an index of enhanced synaptic strength. A recent example is the enlarged excitatory field potentials in the dentate gyrus of rats that are learning spatial relations by exploration. The altered hippocampal field potentials do not reflect learning-specific cellular changes but result from a concomitant rise in brain temperature that is caused by the associated muscular effort. Enhanced dentate field excitatory potentials followed both passive and active heating and were linearly related to the brain temperature. These temperature-related effects may mask any learning-induced changes in field potential.


New insights into handling missing values in environmental epidemiological studies.
Missing data are unavoidable in environmental epidemiologic surveys. The aim of this study was to compare methods for handling large amounts of missing values: omission of missing values, single and multiple imputations (through linear regression or partial least squares regression), and a fully Bayesian approach. These methods were applied to the PARIS birth cohort, where indoor domestic pollutant measurements were performed in a random sample of babies' dwellings. A simulation study was conducted to assess performances of different approaches with a high proportion of missing values (from 50% to 95%). Different simulation scenarios were carried out, controlling the true value of the association (odds ratio of 1.0, 1.2, and 1.4), and varying the health outcome prevalence. When a large amount of data is missing, omitting these missing data reduced statistical power and inflated standard errors, which affected the significance of the association. Single imputation underestimated the variability, and considerably increased risk of type I error. All approaches were conservative, except the Bayesian joint model. In the case of a common health outcome, the fully Bayesian approach is the most efficient approach (low root mean square error, reasonable type I error, and high statistical power). Nevertheless for a less prevalent event, the type I error is increased and the statistical power is reduced. The estimated posterior distribution of the OR is useful to refine the conclusion. Among the methods handling missing values, no approach is absolutely the best but when usual approaches (e.g. single imputation) are not sufficient, joint modelling approach of missing process and health association is more efficient when large amounts of data are missing.


Glacial earthquakes.
We have detected dozens of previously unknown, moderate earthquakes beneath large glaciers. The seismic radiation from these earthquakes is depleted at high frequencies, explaining their nondetection by traditional methods. Inverse modeling of the long-period seismic waveforms from the best-recorded earthquake, in southern Alaska, shows that the seismic source is well represented by stick-slip, downhill sliding of a glacial ice mass. The duration of sliding in the Alaska earthquake is 30 to 60 seconds, about 15 to 30 times longer than for a regular tectonic earthquake of similar magnitude.


Smoking and diabetes.
The objective of this review is to summarize the literature on diabetes and smoking related to epidemiological risks, efficacy and cost-effectiveness of different cessation approaches, and implications for clinical practice. Over 200 studies were reviewed, with special emphasis placed on publications within the past 10 years. Intervention studies that included patients with diabetes but did not report results separately by disease are included. Diabetes-specific studies are highlighted. There are consistent results from both cross-sectional and prospective studies showing enhanced risk for micro- and macrovascular disease, as well as premature mortality from the combination of smoking and diabetes. The general cessation literature is extensive, generally well-designed, and encouraging regarding the impact of cost-effective practical office-based interventions. In particular, system-based approaches that make smoking a routine part of office contacts and provide multiple prompts, advice, assistance, and follow-up support are effective. Although there is minimal information on the effectiveness of cessation interventions specifically for people with diabetes, there is no reason to assume that cessation intervention would be more or less effective in this population. There is a clear need to increase the frequency of smoking cessation advice and counseling for patients with diabetes given the strong and consistent data on smoking prevalence; combined risks of smoking and diabetes for morbidity, mortality, and several complications; and the proven efficacy and cost-effectiveness of cessation strategies.


Differences in behavioral perceptions between young adult burn survivors and cross-informants.
The purpose of this study is to gain insight into the level of agreement, in terms of personal problems, between burn survivors and an informant with whom they have a close personal relationship. Participants in the study were 72 young adult burn survivors between the ages of 18 and 28 with a mean age of 20.9 +/- 2.6 years. There were 37 males and 35 females in the group. The mean total burn surface area was 55.34 +/- 19.9% with injuries occurring before the age of 18, a minimum of 2 years prior to the study with a mean of 14.3 +/- 4.9 years since burned. To assess this issue the participants completed The Young Adult Self-Report (YASR), and the informants completed The Young Adult Behavior Checklist (YABCL). In relationship to burn survivors, the informant group comprised 67% mothers, 3% fathers, and 30% domestic partners. Paired t-test and Pearson Correlations Coefficients were calculated to compare the differences and variance of the Total Problems Scale, Internalizing Scale and Externalizing Scale for the YABCL and YASR results. The family members of burn survivors reported more problems for their young adult relatives than did the informants of the reference population. In general, the YACBL scores were higher than and highly correlated to the YASR scores. Although there were not ethic differences in the YABCL and the YASR measures, the correlation between the YABCL and the YASR was excellent for Caucasian families but not good for Black and Hispanic families. This study supports concerns about the need for routine follow-up into adulthood for those individuals who were burned as children. This would seem to be especially critical for the female population.


The tight junction component Claudin E is required for zebrafish epiboly.
Zebrafish epiboly results in the thinning and spreading of the blastoderm to cover the yolk cell and close the blastopore. The extra-embryonic yolk syncytial layer (YSL) tows the blastoderm vegetally during epiboly by means of its tight junction attachments to the enveloping layer (EVL). Claudins are the major transmembrane protein components of tight junctions. Here, we focus on the function of Claudin E (Cldne), which is expressed specifically in the EVL. Morpholino knock-down of cldne produced a highly penetrant epiboly delay. Our analysis suggested that the EVL margin, which is attached to the YSL, was under reduced tension in morphant embryos. We propose that local variation in the strength of EVL-YSL attachment in morphant embryos resulted in slow and uneven advancement of the EVL and blastoderm. Our work is the first to demonstrate that Claudins are important for zebrafish epiboly.


Surface-assisted diastereoselective Ullmann coupling of bishelicenes.
Ullmann coupling of chiral 2-bromo[4]helicene has been performed on a Cu(100) surface. Only homochiral 2,2'-bis[4]helicene as the product is observed using STM. Such stereoselectivity is based on the fact that the surface will favour a configuration with the central part of the molecule on the surface, causing the outer ends to spiral away from the surface.


Calcium regulation and signaling in apicomplexan parasites.
Apicomplexan parasites rely on calcium-mediated signaling for a variety of vital functions including protein secretion, motility, cell invasion, and differentiation. These functions are controlled by a variety of specialized systems for uptake and release of calcium, which acts as a second messenger, and on the functions of calcium-dependent proteins. Defining these systems in parasites has been complicated by their evolutionary distance from model organisms and practical concerns in working with small, and somewhat fastidious cells. Comparative genomic analyses of Toxoplasma gondii, Plasmodium spp. and Cryptosporidium spp. reveal several interesting adaptations for calcium-related processes in parasites. Apicomplexans contain several P-type Ca2+ ATPases including an ER-type reuptake mechanism (SERCA), which is the proposed target of artemisinin. All three organisms also contain several genes related to Golgi PMR-like calcium transporters, and a Ca2+/H+ exchanger, while plasma membrane-type (PMCA) Ca2+ ATPases and voltage-dependent calcium channels are exclusively found in T. gondii. Pharmacological evidence supports the presence of IP3 and ryanodine channels for calcium-mediated release. Collectively these systems regulate calcium homeostasis and release calcium to act as a signal. Downstream responses are controlled by a family of EF-hand containing calcium binding proteins including calmodulin, and an array of centrin and caltractin-like genes. Most surprising, apicomplexans contain a diversity of calcium-dependent protein kinases (CDPK), which are commonly found in plants. Toxoplasma contains more than 20 CDPK or CDPK-like proteases, while Plasmodium and Cryptosporidium have fewer than half this number. Several of these CDPKs have been shown to play vital roles in protein secretion, invasion, and differentiation, indicating that disruption of calcium-regulated pathways may provide a novel means for selective inhibition of parasites.


Human ALKBH4 interacts with proteins associated with transcription.
The Fe(II)- and 2-oxoglutarate (2OG)-dependent dioxygenase AlkB from E. coli is a demethylase which repairs alkyl lesions in DNA, as well as RNA, through a direct reversal mechanism. Humans possess nine AlkB homologs (ALKBH1-8 and FTO). ALKBH2 and ALKBH3 display demethylase activities corresponding to that of AlkB, and both ALKBH8 and FTO are RNA modification enzymes. The biochemical functions of the rest of the homologs are still unknown. To increase our knowledge on the functions of ALKBH4 and ALKBH7 we have here performed yeast two-hybrid screens to identify interaction partners of the two proteins. While no high-confidence hits were detected in the case of ALKBH7, several proteins associated with chromatin and/or involved in transcription were found to interact with ALKBH4. For all interaction partners, the regions mediating binding to ALKBH4 comprised domains previously reported to be involved in interaction with DNA or chromatin. Furthermore, some of these partners showed nuclear co-localization with ALKBH4. However, the global gene expression pattern was only marginally altered upon ALKBH4 over-expression, and larger effects were observed in the case of ALKBH7. Although the molecular function of both proteins remains to be revealed, our findings suggest a role for ALKBH4 in regulation of gene expression or chromatin state.


Scoping review protocol on research prioritisation for preparedness and response to outbreaks of high consequence pathogens.
Background : Prioritisation of research activities for infectious disease pathogens is usually undertaken through the identification of important research and knowledge gaps. Research prioritisation is an essential element of both effective responses to disease outbreaks and adequate preparedness. There is however currently no published mapping of activities on and evidence from research prioritisation for high consequence pathogens. The objectives of this review are to map all published research prioritisation exercises on high-consequence pathogens; provide an overview of methodologies employed for prioritising research for these pathogens; describe monitoring and evaluation processes for research areas prioritised; and identify any standards and guidance for effectively undertaking research prioritisation activities for high consequence pathogens. Methods: The Joanna Briggs Institute guidance of scoping review conduct will be used. The search will be undertaken using the key terms of "research prioritisation", "response", "control", and related terms, and a list of high-consequence pathogens derived from WHO (2020), EMERGE (2019), Europe CDC (2022) and the Association of Southeast Asian Nations (2021). We will search WHO Global Index Medicus; Ovid Medline; Ovid Embase; Ovid Global Health; and Scopus. Backward citations review of the included full text documents will also be conducted. Google Scholar and Overton will be searched for grey literature. Two independent reviewers will screen the retrieved documents using Rayyan and extract data in a data extraction template in Microsoft Excel 2021. Screening results will be presented using the PRISMA-ScR template with narrative synthesis undertaken for the extracted data. Conclusion: This review will map existing research priorities for high consequence pathogens. Further, it will provide an understanding of methodologies used for prioritisation, processes for monitoring and evaluation of progress made against research agendas, and evidence on standards that could be recommended for effective prioritisation of research for high consequence pathogens.


Apvasa marks germ-cell migration in the parthenogenetic pea aphid Acyrthosiphon pisum (Hemiptera: Aphidoidea).
In the parthenogenetic and viviparous pea aphid Acyrthosiphon pisum, germline specification depends on the germ plasm localized to the posterior region of the egg chamber before the formation of the blastoderm. During blastulation, germline segregation occurs at the egg posterior, and in early gastrulation germ cells are pushed inward by the invaginating germ band. Previous studies suggest that germ cells remain dorsal in the embryo in subsequent developmental stages. In fact, though, it is not known whether germ cells remain in place or migrate dynamically during katatrepsis and germ-band retraction. We cloned Apvasa, a pea aphid homologue of Drosophila vasa, and used it as a germline marker to monitor the migration of germ cells. Apvasa messenger RNA (mRNA) was first restricted to morphologically identifiable germ cells after blastoderm formation but that expression soon faded. Apvasa transcripts were again identified in germ cells from the stage when the endosymbiotic bacteria invaded the embryo, and after that, Apvasa mRNA was present in germ cells throughout all developmental stages. At the beginning of katatrepsis, germ cells were detected at the anteriormost region of the egg chamber as they were migrating into the body cavity. During the early period of germ-band retraction, germ cells were separated into several groups surrounded by a layer of somatic cells devoid of Apvasa staining, suggesting that the coalescence between migrating germ cells and the somatic gonadal mesoderm occurs between late katatrepsis and early germ-band retraction.


Response variability to glucose facilitation of cognitive enhancement.
Glucose facilitation of cognitive function has been widely reported in previous studies (including our own). However, several studies have also failed to detect glucose facilitation. There is sparsity of research examining the factors that modify the effect of glucose on cognition. The aims of the present study were to (1) demonstrate the previously observed enhancement of cognition through glucose administration and (2) investigate some of the factors that may exert moderating roles on the behavioural response to glucose, including glucose regulation, body composition (BC) and hypothalamic–pituitary–adrenal axis response. A total of twenty-four participants took part in a double-blind, placebo-controlled, randomised, repeated-measures study, which examined the effect of 25 and 60 g glucose compared with placebo on cognitive function. At 1 week before the study commencement, all participants underwent an oral glucose tolerance test. Glucose facilitated performance on tasks of numeric and spatial working memory, verbal declarative memory and speed of recognition. Moderating variables were examined using several indices of glucoregulation and BC. Poorer glucoregulation predicted improved immediate word recall accuracy following the administration of 25 g glucose compared with placebo. Those with better glucoregulation showed performance decrements on word recall accuracy following the administration of 25 g glucose compared with placebo. These findings are in line with accumulating evidence that glucose load may preferentially enhance cognition in those with poorer glucoregulation. Furthermore, the finding that individuals with better glucoregulation may suffer impaired performance following a glucose load is novel and requires further substantiation.


Can recall of birth size be used as a measure of birthweight in Cameroon?
Birthweight is a key predictor of future life outcomes. When this information is not available we must rely on maternal recall of weight or size. We evaluated the use of maternal recall of size as an indicator for birthweight, using data from the 2004 Demographic and Health Survey in Cameroon. The overall agreement between recorded and recalled weight and subjective assessment of birth size was 90% (Kappa = 0.46). Concordant descriptions were associated with higher birthweights (P < 0.001), higher education (P = 0.008) and delivery in a health unit (P = 0.025). Specificity for low birthweight (92.9%) was much higher than sensitivity (59.9%) and the negative predictive value (96.1%) was much higher than the positive predictive value (44.4%). These results indicate that recall of size, in Cameroonian women and in other low resource settings, should be used only in the absence of other sources of data.


Quantification of bone strontium levels in humans by in vivo x-ray fluorescence.
The need for in vivo bone strontium assessment arises because strontium may exert a number of effects on bone, which may be either beneficial or toxic. Measurements discussed here are noninvasive, no sample is taken, nor is there discomfort to patients. The developed source excited x-ray fluorescence system employs a 109Cd source to excite the strontium K x rays, with the source and detector in approximately 90 degree geometry relative to the sample position. The factors affecting the accuracy and minimal detectable limit for bone strontium in vivo measurements are discussed. A system calibration revealed a minimum detectible limit of approximately 0.25 mg Sr/g Ca, which is sufficient for the monitoring of strontium levels in healthy subjects and patients with elevated bone strontium concentrations. Preliminary in vivo measurements in ten healthy subjects at two bone sites (phalanx and tibia) indicated that this system can be applied for cumulative bone strontium estimation while delivering a low effective dose of 80 nSv during the measurement time. Future work will involve attempts to enhance system precision with alternative fluorescing sources and further optimization of the detection system.


Effects of Social Defeat Stress on Sleep in Mice.
Stress plays a key role in the development of psychiatric disorders and has a negative impact on sleep integrity. In mice, chronic social defeat stress (CSDS) is an ethologically valid model of stress-related disorders but little is known about its effects on sleep regulation. Here, we investigated the immediate and long-term effects of 10 consecutive days of social defeat (SD) on vigilance states in C57Bl/6J male mice. Social behavior was assessed to identify susceptible mice, i.e., mice that develop long-lasting social avoidance, and unsusceptible mice. Sleep-wake stages in mice of both groups were analyzed by means of polysomnographic recordings at baseline, after the first, third, and tenth stress sessions and on the 5th recovery day (R5) following the 10-day CSDS. In susceptible mice, each SD session produced biphasic changes in sleep-wake states that were preserved all along 10-day CSDS. These sessions elicited a short-term enhancement of wake time while rapid eye-movement (REM) sleep was strongly inhibited. Concomitantly, delta power was increased during non REM (NREM) sleep. During the following dark period, an increase in total sleep time, as well as wake fragmentation, were observed after each analyzed SD session. Similar changes were observed in unsusceptible mice. At R5, elevated high-frequency EEG activity, as observed in insomniacs, emerged during NREM sleep in both susceptible and unsusceptible groups suggesting that CSDS impaired sleep quality. Furthermore, susceptible but not unsusceptible mice displayed stress-anticipatory arousal during recovery, a common feature of anxiety disorders. Altogether, our findings show that CSDS has profound impacts on vigilance states and further support that sleep is tightly regulated by exposure to stressful events. They also revealed that susceptibility to chronic psychological stress is associated with heightened arousal, a physiological feature of stress vulnerability.