Stable and sporadic symbiotic communities of coral and algal holobionts.
Coral and algal holobionts are assemblages of macroorganisms and microorganisms, including viruses, Bacteria, Archaea, protists and fungi. Despite a decade of research, it remains unclear whether these associations are spatial-temporally stable or species-specific. We hypothesized that conflicting interpretations of the data arise from high noise associated with sporadic microbial symbionts overwhelming signatures of stable holobiont members. To test this hypothesis, the bacterial communities associated with three coral species (Acropora rosaria, Acropora hyacinthus and Porites lutea) and two algal guilds (crustose coralline algae and turf algae) from 131 samples were analyzed using a novel statistical approach termed the Abundance-Ubiquity (AU) test. The AU test determines whether a given bacterial species would be present given additional sampling effort (that is, stable) versus those species that are sporadically associated with a sample. Using the AU test, we show that coral and algal holobionts have a high-diversity group of stable symbionts. Stable symbionts are not exclusive to one species of coral or algae. No single bacterial species was ubiquitously associated with one host, showing that there is not strict heredity of the microbiome. In addition to the stable symbionts, there was a low-diversity community of sporadic symbionts whose abundance varied widely across individual holobionts of the same species. Identification of these two symbiont communities supports the holobiont model and calls into question the hologenome theory of evolution.


Experimental investigation of pulse generation with one-pump fiber optical parametric amplification.
In a recent study, the theory of pulse generation with fiber optical parametric amplification using sinusoidal (clock) intensity modulated pump was revisited. This work showed that the pulses generated through such parametric interaction exhibit a shape which depends on the signal detuning with respect to the pump position (i.e. linear phase mismatch). A near Gaussian shape can only be achieved over a small region of the gain spectrum, close to the maximum gain location. Towards the extremities of the gain spectrum, the generated pulses take a near Sinc shape which can have many potential applications such as for all-optical Nyquist limited transmitters and/or receivers. In this paper we experimentally verify the theory at repetition rates up to 40 GHz. We also discuss the impact of noise, pump saturation and walk-off on the generated pulses.


"My wife's mistrust. That's the saddest part of being a diabetic": A qualitative study of sexual well-being in men with Type 2 diabetes in sub-Saharan Africa.
Sexual dysfunction is a common complication for men with diabetes, yet little is known about the lived experiences of sexual difficulties within the context of diabetes, particularly in low-and-middle-income countries. This study explores how men with type 2 diabetes in three sub-Saharan African settings (Cape Town and Johannesburg, South Africa; Lilongwe, Malawi) perceive and experience sexual functioning and sexual well-being, and the biopsychosocial contexts in which these occur and are shaped. We used a qualitative research design, including individual interviews (n = 15) and focus group discussions (n = 4). Forty-seven men were included in the study. We used an inductive thematic analysis approach to develop our findings. A biopsychosocial conceptual model on the relationship between chronic illness and sexuality informed the interpretation of findings. Men across the study settings identified sexual difficulties as a central concern of living with diabetes. These difficulties went beyond biomedical issues of erectile dysfunction, comprising complex psychological and relational effects. Low self-esteem, related to a sense of loss of masculinity and reduced sexual and emotional intimacy in partner relationships were common experiences. Specific negative relational effects included suspicion of infidelity, mutual mistrust, general unhappiness, and fear of losing support from partners. These effects may impact on men's ability to cope with their diabetes. Further stressors were a lack of information about the reasons for their sexual difficulties, perceived lack of support from healthcare providers and an inability to communicate with partners about sexual difficulties. More in-depth research is needed to better understand sexual functioning and well-being within the context of diabetes, and its potential impact on diabetes self-management. Holistic and patient-centered care should include raising awareness of sexual problems as a potential complication of diabetes amongst patients, their partners and care providers, and incorporating sexual well-being as part of routine clinical care.


To what extent do spinal interactions between an alpha-2 adrenoceptor agonist and a mu opioid agonist influence noxiously evoked c-Fos expression in the rat? A pharmacological study.
Three hours after the intraplantar injection of carrageenin (6 mg/150 microliters of saline) Fos-like immunoreactivity (Fos-LI) was observed in both superficial and deep laminae of the dorsal horn segments L4 and L5 of the spinal cord. Systemic medetomidine, an alpha-2 adrenoceptor agonist (12.5, 25 or 75 micrograms/kg i.v.), dose-dependently reduced the number of superficial and deep Fos-LI neurons; 75 micrograms/kg produced a 66 +/- 4% and a 90 +/- 4% reduction of superficial and deep Fos-LI neurons, respectively, P < .0001 for both. In addition, systemic medetomidine dose-relatedly reduced the carrageenin-evoked paw and ankle edema; medetomidine 75 micrograms/kg resulted in a 70 +/- 3% reduction of paw edema and in a blockade of the development of ankle edema. The effects of medetomidine were blocked by systemic atipamezole (75 micrograms/kg, i.v.), which, when injected alone, had no effect on the number of Fos-LI neurons or the peripheral edema. Co-administration of a low dose of medetomidine (12.5 micrograms/kg i.v.) with an ineffective dose of morphine (1.5 micrograms/kg i.v.) strongly decreased the number of superficial and deep Fos-LI neurons (40 +/- 5%, P < .0001 and 62 +/- 11%, P < .0001 reduction as compared with control group) without altering the effects of medetomidine on the peripheral edema. Both atipamezole and a combined injection of atipamezole and naloxone blocked the effects of medetomidine plus morphine on both the total number of Fos-LI neurons (86 +/- 11% and 86 +/- 6% of control, respectively) and carrageenin inflammation (87 +/- 6%, P < .05 and 84 +/- 3%, P < .05 of control for the paw edema; 75 +/- 8%, P < .01 and 81 +/- 7%, P < .05 of control for the ankle edema, respectively). Naloxone alone blocked the effects of the co-administered agonists on the total number of Fos-Li neurons (91 +/- 6% of the control carrageenin group) without influencing the effect on the peripheral edema. Our results demonstrate, for the first time, that co-administration of alpha-2 adrenoceptor and mu opioid agonists substantially reduces inflammatory evoked expression of c-Fos, one of the long-term consequences of sustained nociceptive processing.


The RF-powered surface wave sensor oscillator--a successful alternative to passive wireless sensing.
A novel, passive wireless surface acoustic wave (SAW) sensor providing a highly coherent measurand proportional frequency, frequency modulated (FM) with identification (ID) data and immune to interference with multiple-path signals is described. The sensor is appropriate for bandwidth-limited applications requiring high-frequency accuracy. It comprises a low-power oscillator, stabilized with the sensing SAW resonator and powered by the rectified radio frequency (RF) power of the interrogating signal received by an antenna on the sensor part. A few hundred microwatts of direct current (DC) power are enough to power the sensor oscillator and ID modulation circuit and achieve stable operation at 1.0 and 2.49 GHz. Reliable sensor interrogation was achieved over a distance of 0.45 m from a SAW-based interrogation unit providing 50 mW of continuous RF power at 915 MHz. The -30 to -35 dBm of returned sensor power was enough to receive the sensor signal over a long distance and through several walls with a simple superheterodyne FM receiver converting the sensor signal to a low measurand proportional intermediate frequency and retrieving the ID data through FM detection. Different sensor implementations, including continuous and pulsed power versions and the possibility of transmitting data from several measurands with a single sensor, are discussed.


Has the great recession and its aftermath reduced traffic fatalities?
An analysis of state-level data from 1984 to 2014 provides evidence on the relationship between economic recessions and US traffic fatalities. While there are large reductions associated with decreases in household median income, other policy variables tend to have additional and in some cases, larger effects. An increase in the inequality of the income distribution, measured by the Gini index, has reduced traffic fatalities. Graduated licensing policies, cell phone laws, and motorcycle helmet requirements are all associated with reductions in fatalities. Other factors include a proxy for medical technology, and access to emergency medical services (based on the percent of vehicle miles traveled in rural areas); reductions in the latter accounted for a substantial reduction in fatalities and is likely another indicator of reduced economic activity. Changes in the road network, mainly increases in the percent of collector roads has increased fatalities. Population growth is associated with increased traffic fatalities and changes in age cohorts has a small negative effect. Overall, results suggest that there has been a beneficial impact on traffic fatalities from reduced economic activity, but various policies adopted by the states have also reduced traffic fatalities.


Genome Plasticity of agr-Defective Staphylococcus aureus during Clinical Infection.
Therapy for bacteremia caused by Staphylococcus aureus is often ineffective, even when treatment conditions are optimal according to experimental protocols. Adapted subclones, such as those bearing mutations that attenuate agr-mediated virulence activation, are associated with persistent infection and patient mortality. To identify additional alterations in agr-defective mutants, we sequenced and assembled the complete genomes of clone pairs from colonizing and infected sites of several patients in whom S. aureus demonstrated a within-host loss of agr function. We report that events associated with agr inactivation result in agr-defective blood and nares strain pairs that are enriched in mutations compared to pairs from wild-type controls. The random distribution of mutations between colonizing and infecting strains from the same patient, and between strains from different patients, suggests that much of the genetic complexity of agr-defective strains results from prolonged infection or therapy-induced stress. However, in one of the agr-defective infecting strains, multiple genetic changes resulted in increased virulence in a murine model of bloodstream infection, bypassing the mutation of agr and raising the possibility that some changes were selected. Expression profiling correlated the elevated virulence of this agr-defective mutant to restored expression of the agr-regulated ESAT6-like type VII secretion system, a known virulence factor. Thus, additional mutations outside the agr locus can contribute to diversification and adaptation during infection by S. aureus agr mutants associated with poor patient outcomes.


Innovative method for recovery and valorization of hydroxytyrosol from olive mill wastewaters.
The nutritional properties of olive oil can be attributed to its oleic acid and phenolic compounds content, acting as natural oxidants to prevent human diseases. In particular, hydroxytyrosol has an anti-inflammatory action similar to omega 3 fatty acids from fish oil. The olive oil production was conducted by two extraction procedures: first, a two-phase extraction giving extra-virgin olive oil and humid pomace, second, a three-phase working process of humid pomace, obtaining another minimum quantity of extra-virgin olive oil, 'dry' pomace devoid of polyphenols, and mill wastewaters rich in anti-oxidant compounds. The aim of this processing was to employ water to extract the highest concentration of polyphenols from humid pomace and convey them in oil mill wastewaters for extraction. Processed olives were 37,200 kg, pomace deprived of polyphenols was equal to 20,400 kg and processing was performed with 500 kg of olives per hour. This method offers advantages of using cheap equipment and technical simplicity.


Physical and neurodevelopmental outcomes in children with single-ventricle circulation.
To investigate longer-term physical and neurodevelopmental outcomes of patients with hypoplastic left heart syndrome (HLHS) compared with other patients with functionally single-ventricle circulation surviving beyond the age of 10 years. A retrospective, observational study from a UK tertiary centre for paediatric cardiology. 58 patients with HLHS and 44 non-HLHS patients with single-ventricle physiology were included. Subjective reduction in exercise tolerance was reported in 72% (95% CI 61% to 84%) of patients with HLHS and 45% (31% to 60%) non-HLHS patients. Compared with non-HLHS patients, educational concerns were reported more frequently in patients with HLHS, 41% (29% to 54%) vs 23% (10% to 35%), as was a diagnosis of a behaviour disorder (autism or attention deficit hyperactivity disorder) 12% (4% to 21%) vs 0%, and referral to other specialist services 67% (55% to 79%) vs 48% (33% to 63%). Within a group of young people with complex congenital heart disease, those with HLHS are likely to have worse physical, psychological and educational outcomes.


The blood-cerebrospinal fluid barrier--first evidence for an active transport of organic mercury compounds out of the brain.
Exposure to organic mercury compounds promotes primarily neurological effects. Although methylmercury is recognized as a potent neurotoxicant, its transfer into the central nervous system (CNS) is not fully evaluated. While methylmercury and thiomersal pass the blood-brain barrier, limited data are available regarding the second brain regulating interface, the blood-cerebrospinal fluid (CSF) barrier. This novel study was designed to investigate the effects of organic as well as inorganic mercury compounds on, and their transfer across, a porcine in vitro model of the blood-CSF barrier for the first time. The barrier system is significantly more sensitive towards organic Hg compounds as compared to inorganic compounds regarding the endpoints cytotoxicity and barrier integrity. Whereas there are low transfer rates from the blood side to the CSF side, our results strongly indicate an active transfer of the organic mercury compounds out of the CSF. These results are the first to demonstrate an efflux of organic mercury compounds regarding the CNS and provide a completely new approach in the understanding of mercury compounds specific transport.


Whole genome resequencing identifies local adaptation associated with environmental variation for redband trout.
Aquatic ectotherms are predicted to harbour genomic signals of local adaptation resulting from selective pressures driven by the strong influence of climate conditions on body temperature. We investigated local adaptation in redband trout (Oncorhynchus mykiss gairdneri) using genome scans for 547 samples from 11 populations across a wide range of habitats and thermal gradients in the interior Columbia River. We estimated allele frequencies for millions of single nucleotide polymorphism loci (SNPs) across populations using low-coverage whole genome resequencing, and used population structure outlier analyses to identify genomic regions under divergent selection between populations. Twelve genomic regions showed signatures of local adaptation, including two regions associated with genes known to influence migration and developmental timing in salmonids (GREB1L, ROCK1, SIX6). Genotype-environment association analyses indicated that diurnal temperature variation was a strong driver of local adaptation, with signatures of selection driven primarily by divergence of two populations in the northern extreme of the subspecies range. We also found evidence for adaptive differences between high-elevation desert vs. montane habitats at a smaller geographical scale. Finally, we estimated vulnerability of redband trout to future climate change using ecological niche modelling and genetic offset analyses under two climate change scenarios. These analyses predicted substantial habitat loss and strong genetic shifts necessary for adaptation to future habitats, with the greatest vulnerability predicted for high-elevation desert populations. Our results provide new insight into the complexity of local adaptation in salmonids, and important predictions regarding future responses of redband trout to climate change.


Gestational diabetes may represent discovery of preexisting glucose intolerance.
A representative sample of 817 American women aged 20-44 yr who were not pregnant and had no medical history of diabetes were given 75-g 2-h oral glucose tolerance tests (OGTTs). Although these conditions are somewhat different from those recommended for pregnant women (100 g glucose, 3-h OGTT), 3.8% of the women might have been considered to have met O'Sullivan and Mahan criteria for gestational diabetes mellitus (GDM) had they been pregnant. Prevalence was 2-3% below age 35 yr, similar to that found in studies of pregnant women, and rose to 8% at age 40-44 yr. Rates of women meeting World Health Organization criteria for gestational impaired glucose tolerance (G-IGT) rose steadily from 5% at age 20-24 yr to 11% at age 40-44 yr. Risk factors for non-insulin-dependent diabetes mellitus (NIDDM) including parental history and obesity were more prevalent among women meeting these criteria than among women in the entire group; the same risk factors are also more prevalent among pregnant women with GDM. The similarity of rates in this study to rates of GDM and G-IGT, together with their association with risk factors for NIDDM, indicate that these entities are compatible with undiagnosed glucose intolerance occurring before pregnancy and discovered during the metabolic testing that generally accompanies prenatal care rather than conditions that have an etiologic relationship to pregnancy.


Indoleamine-pyrrole 2,3-dioxygenase-1 (IDO-1) mRNA is over-expressed in the duodenal mucosa and is negatively correlated with serum tryptophan concentrations in dogs with protein-losing enteropathy.
Dogs with protein-losing enteropathy (PLE) have decreased serum tryptophan concentrations, which may contribute to disease pathogenesis. Indoleamine-pyrrole 2,3-dioxygenase-1 (IDO-1) expression is associated with low serum tryptophan concentrations and is increased in the gastrointestinal tract of humans with inflammatory bowel disease (IBD). Therefore, the objective of our study was to determine if the mRNA expression of IDO-1 is increased in the duodenal mucosa of dogs with PLE as compared to dogs with chronic enteropathy (CE) and healthy dogs, and whether this expression is correlated with changes in serum tryptophan concentration. Our study was a retrospective study using archived paraffin-embedded duodenal biopsy specimens from 8 healthy Beagle dogs from the Iowa State University Canine Service Colony and 18 and 6 client-owned dogs diagnosed with CE and PLE, respectively at the Bristol Veterinary School. A novel RNA in situ hybridization (ISH) technology, RNAscope, was used to identify IDO-1 mRNA mucosal expression in duodenal tissues. An IDO-1 specific probe was hybridized onto 10 duodenal biopsy sections from each dog whereby RNAscope signal (mRNA expression) was quantified by a single operator using light microscopy. Dogs with PLE had significantly higher mRNA expression of IDO-1 in the duodenal mucosa compared to healthy dogs (mucosal percentage IDO-1 positive: P = 0.0093, (mean ± S.D) control: 19.36 ± 7.08, PLE: 34.12 ± 5.98, average fold difference: 1.76 and mucosal IDO-1 H-score: P = 0.0356, (mean ± S.D) control: 45.26 ± 19.33, PLE: 84.37 ± 19.86, average fold difference: 1.86). The duodenal mucosal mRNA expression of IDO-1 was negatively correlated with serum tryptophan concentrations in dogs with PLE (mucosal IDO-1 H-score: Spearman's rank correlation coefficient = -0.94, P = 0.0048). In conclusion, our study suggests that decreased serum tryptophan concentrations in dogs with PLE is associated with increased intestinal IDO-1 expression. Further studies are needed to determine potential inflammatory pathways responsible for increased expression of IDO-1 in the intestinal tract of dogs with PLE.


Development and Validation of a Clinical Prognostic Model Based on Immune-Related Genes Expressed in Clear Cell Renal Cell Carcinoma.
Background: Clear cell renal cell carcinoma (ccRCC) is the most frequent and terminal subtype of RCC. Reliable markers associated with the immune response are not available to predict the prognosis of patients with ccRCC. We exploited the extensive number of ccRCC samples from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) repository to perform a comprehensive analysis of immune-related genes (IRGs). Methods: Based on TCGA data, we incorporated IRGs and their expression profiles of 72 normal and 539 ccRCC samples. Univariate Cox analysis was used to evaluate the relationship between overall survival (OS) and IRGs expression. The Lasso Cox regression model identified prognostic genes used to establish a clinical immune prognostic model. The TF-IRG network was used to study the potential molecular mechanisms of action and properties of ccRCC-specific IRGs. Multivariate Cox analysis established a clinical prognostic model of IRGs. Results: We found a significant correlation among 15 differentially expressed IRGs with the OS of patients with ccRCC. Gene function enrichment analysis showed that these IRGs are significantly associated with response to receptor ligand activity. Lasso Cox regression analysis identified 10 genes with the greatest prognostic value. A clinical prognostic model based on six IRGs, which performed well for predicting prognosis, revealed significant associations of patients' survival with age, sex, stage, tumor, node, and metastasis. Moreover, these findings reflect the infiltration of tumors by various immune cells. Conclusion: We identified six clinically significant IRGs and incorporated them into a clinical prognostic model with great significance for monitoring and predicting prognosis of ccRCC.


Mass spectrometry of levitated droplets by thermally unconfined infrared-laser desorption.
An ionization scheme for fast online mass spectrometric interrogation of levitated droplets is presented. That renewed method comprises the output of an a Er:YAG laser at λ = 2.94 μm which is in resonance with the OH stretch vibration band of solvents like water and alcohols. A temporal pulse width larger than the time needed for pressure redistribution and also above the temperature redistribution time constant was found to lead to soft evaporation/ionization. Despite these mild desorption conditions, no additional postionization is found to be needed. Accordingly, the ionization is found to be very soft resulting in entirely intact analyte ions and concentration dependent cluster ions. Resulting mass spectra of small amino acids and large antibiotics are presented showing the versatility of the introduced technique. Above a critical mass of m ≈ 1 kDa, the formed ions carry multiple charges as it is typical for thermospray or electrospray ionization. The detection technique enables fast contactless analysis of the chemical composition of levitated microreactors and, thus, paves the way for future contactless reaction monitoring.


Physicochemical and sensory (aroma and colour) characterisation of a non-centrifugal cane sugar ("panela") beverage.
Non-centrifugal cane sugar (NCS), also called "panela", is a high carbohydrate-content food obtained by boil evaporation of the sugar cane juice. This study was undertaken to assess physicochemical properties and sensory characteristics of panela beverage at two different concentrations. Evaluation of pH, °Brix, and colour (tristimulus colorimetry) was carried out in all panela drink samples. In order to characterise the odour-active volatiles of the beverage, a simultaneous steam distillation-solvent extraction method was applied using a mixture of diethyl ether-pentane (1:1,w/w) as solvent. The Aroma Extract Dilution Analysis revealed the presence of six odour-active compounds, being 2-methyl pyrazine the key aroma compound of this beverage. PCA (Principal Component Analysis) showed that there were no differences in the aroma and physicochemical properties (pH and °Brix) with respect to the geographical origin of analysed samples; however colour depends on heating during processing of NCS.


Both epsilon-toxin and beta-toxin are important for the lethal properties of Clostridium perfringens type B isolates in the mouse intravenous injection model.
Clostridium perfringens is capable of producing up to 15 toxins, including alpha-toxin (CPA), beta-toxin (CPB), epsilon-toxin (ETX), enterotoxin, beta2-toxin (CPB2), and perfringolysin O. Type B isolates, which must produce CPA, CPB, and ETX, are associated with animal illnesses characterized by sudden death or acute neurological signs, with or without intestinal damage. Type B pathogenesis in ruminants is poorly understood, with some animals showing lesions and clinical signs similar to those caused by either type C or type D infections. It is unknown whether host or environmental conditions are dominant for determining the outcome of type B disease or if disease outcomes are determined by variable characteristics of type B isolates. To help clarify this issue, 19 type B isolates were evaluated for toxin production during late-log-phase growth via quantitative Western blotting and by biological activity assays. Most type B isolates produced CPB levels similar to those produced by type C isolates in vitro and have the potential to produce genotype C-like disease. The lethality of type B isolate supernatants administered intravenously to mice was evaluated with or without prior trypsin treatment, and monoclonal antibody neutralization studies also were performed. Correlation analyses comparing toxin levels in type B supernatants versus lethality and neutralization studies both found that the main contributor to lethality without pretreatment with trypsin was CPB, whereas neutralization studies indicated that CPB and ETX were both important after trypsin pretreatment. At least part of the CPB produced by type B isolates remained active after trypsin treatment. However, the overall lethalities of most supernatants were lower after trypsin pretreatment. Also, there was a significant association between ETX, CPB2, and CPA production in vitro among type B isolates. However, our results suggest that both CPB and ETX are likely the most important contributors to the pathogenesis of C. perfringens type B infections in domestic animals.


Appendectomy Versus Observation for Appendicitis in Neutropenic Children With Cancer.
Optimal management of neutropenic appendicitis (NA) in children undergoing cancer therapy remains undefined. Management strategies include upfront appendectomy or initial nonoperative management. We aimed to characterize the effect of management strategy on complications and length of stay (LOS) and describe implications for chemotherapy delay or alteration. Sites from the Pediatric Surgery Oncology Research Collaborative performed a retrospective review of children with NA over a 6-year period. Sixty-six children, with a median age of 11 years (range 1-17), were identified with NA while undergoing cancer treatment. The most common cancer diagnoses were leukemia (62%) and brain tumor (12%). Upfront appendectomy was performed in 41% of patients; the remainder had initial nonoperative management. Rates of abscess or perforation at diagnosis were equivalent in the groups (30% vs 24%; P = .23). Of patients who had initial nonoperative management, 46% (17 of 37) underwent delayed appendectomy during the same hospitalization. Delayed appendectomy was due to failure of initial nonoperative management in 65% (n = 11) and count recovery in 35% (n = 6). Cancer therapy was delayed in 35% (n = 23). Initial nonoperative management was associated with a delay in cancer treatment (46% vs. 22%, P = .05) and longer LOS (29 vs 12 days; P = .01). Patients who had initial nonoperative management and delayed appendectomy had a higher rate of postoperative complications (P < .01). In pediatric patients with NA from oncologic treatment, upfront appendectomy resulted in lower complication rates, reduced LOS, and fewer alterations in chemotherapy regimens compared to initial nonoperative management.


De novo transcriptome assembly of a fern, Lygodium japonicum, and a web resource database, Ljtrans DB.
During plant evolution, ferns originally evolved as a major vascular plant with a distinctive life cycle in which the haploid and diploid generations are completely separated. However, the low level of genetic resources has limited studies of their physiological events, as well as hindering research on the evolutionary history of land plants. In this study, to identify a comprehensive catalog of transcripts and characterize their expression traits in the fern Lygodium japonicum, nine different RNA samples isolated from prothalli, trophophylls, rhizomes and sporophylls were sequenced using Roche 454 GS-FLX and Illumina HiSeq sequencers. The hybrid assembly of the high-quality 454 GS-FLX and Illumina HiSeq reads generated a set of 37,830 isoforms with an average length of 1,444 bp. Using four open reading frame (ORF) predictors, 38,142 representative ORFs were identified from a total of 37,830 transcript isoforms and 95 contigs, which were annotated by searching against several public databases. Furthermore, an orthoMCL analysis using the protein sequences of L. japonicum and five model plants revealed various sets of lineage-specific genes, including those detected among land plant lineages and those detected in only L. japonicum. We have also examined the expression patterns of all contigs/isoforms, along with the life cycle of L. japonicum, and identified the tissue-specific transcripts using statistical expression analyses. Finally, we developed a public web resource, the L. japonicum transcriptome database at http://bioinf.mind.meiji.ac.jp/kanikusa/, which provides important opportunities to accelerate molecular research in ferns.


A single nanotrench in a palladium microwire for hydrogen detection.
The hydrogen sensing characteristics of a single nanotrench fabricated by focused ion beam milling (FIB) in an evaporated palladium microwire are presented. In situ atomic force microscopy (AFM) measurements proved that, in the presence of H(2), the trench closes and electrically connects the initially separated parts of the wire due to the increase in volume of the material. Therewith, an electrical current can be switched through the wire. With experiments under various H(2) concentrations and a mathematical model, we describe the closing mechanism of the trench with respect to various parameters, including the substrate material, film thickness, trench size and wire dimensions. Results have been compared with those from equivalent continuous wires. Thin SiO(2) and polyimide (PI) layers on silicon were used to study the effect of substrate elasticity. Sufficient lateral expansion of Pd to close trenches of up to 70 nm in width has only been observed on PI, which we attribute to its advantageous elastic properties. The scale of the response times allowed the observation of two superposing effects: the chemical conversion of Pd to PdH(x) and the mechanical closing of the trench.


Temporary structures as a generator of waste in covered trade fairs.
Events like trade fairs are a complex service activity with a considerable economic, social and environmental impact due, among other factors, to their high level of waste generation. There are few studies of the environmental impact associated with waste generation and typology. An environmental analysis methodology has been developed to characterise the waste associated with the temporary structures used at trade fair events: stands and communal spaces. This methodology has been checked in a pilot test at 6 closed trade fairs in Barcelona, with a range of between 60 and 4400 exhibitors. The methodology developed has made possible to obtain a waste generation profile according to the size of the fair and the types of stands. The stages with the largest amount of temporary structure wastes generated are the assembly and the dismantling of the trade fair. The results indicate that the most common wastes generated are the protective plastic from carpets at the assembly stage and the carpet itself at the dismantling stage. The stand carpet is collected in bulk, while the carpet from the communal spaces is recycled. As the size of the fair increases, and with it the proportion of stands with customised design (or non-reusable stands), the quantity of wood and hazardous waste increases.


Maternal near miss and mortality in a rural referral hospital in northern Tanzania: a cross-sectional study.
Maternal morbidity and mortality in sub-Saharan Africa remains high despite global efforts to reduce it. In order to lower maternal morbidity and mortality in the immediate term, reduction of delay in the provision of quality obstetric care is of prime importance. The aim of this study is to assess the occurrence of severe maternal morbidity and mortality in a rural referral hospital in Tanzania as proposed by the WHO near miss approach and to assess implementation levels of key evidence-based interventions in women experiencing severe maternal morbidity and mortality. A prospective cross-sectional study was performed from November 2009 until November 2011 in a rural referral hospital in Tanzania. All maternal near misses and maternal deaths were included. As not all WHO near miss criteria were applicable, a modification was used to identify cases. Data were collected from medical records using a structured data abstraction form. Descriptive frequencies were calculated for demographic and clinical variables, outcome indicators, underlying causes, and process indicators. In the two-year period there were 216 maternal near misses and 32 maternal deaths. The hospital-based maternal mortality ratio was 350 maternal deaths per 100,000 live births (95% CI 243-488). The maternal near miss incidence ratio was 23.6 per 1,000 live births, with an overall case fatality rate of 12.9%. Oxytocin for prevention of postpartum haemorrhage was used in 96 of 201 women and oxytocin for treatment of postpartum haemorrhage was used in 38 of 66 women. Furthermore, eclampsia was treated with magnesium sulphate in 87% of all cases. Seventy-four women underwent caesarean section, of which 25 women did not receive prophylactic antibiotics. Twenty-eight of 30 women who were admitted with sepsis received parenteral antibiotics. The majority of the cases with uterine rupture (62%) occurred in the hospital. Maternal morbidity and mortality remain challenging problems in a rural referral hospital in Tanzania. Key evidence-based interventions are not implemented in women with severe maternal morbidity and mortality. Progress can be made through up scaling the use of evidence-based interventions, such as the use of oxytocin for prevention and treatment of postpartum haemorrhage.


Evaluation of anti-cancer and immunomodulatory effects of carnosol in a Balb/c WEHI-164 fibrosarcoma model.
Agents that destroy tumor cells and simultaneously boost host anti-tumor immunity are of keen interest in cancer therapy. In the present study, the effect of carnosol on anti-tumor immunity in a Balb/c mouse model of fibrosarcoma was evaluated. Carnosol was administered intraperitoneally daily (at 5 or 10 mg/kg/day, for 7 days) to tumor-bearing mice (i.e. 7 days after initial injection of tumor cells). Another group of tumor-bearing mice was treated with 20 mg cyclophosphamide/kg/d (positive control); a final group received vehicle only (vehicle control). After an initial measure on Day 0, tumor size was measured twice during the 7-day treatment period. One day after the final treatment with vehicle/carnosol (i.e. Day 7), the mice had their tumors measured and then were euthanized to permit their spleen and tumor to be harvested for isolation of, respectively, splenocytes and tumor-associated lymphocytes. Using these materials, spontaneous and mitogen-induced release of interleukin (IL)-4, IL-10, and interferon (IFN)-γ, lymphocyte proliferation, and the absolute numbers/relative percentages of splenic and tumor-associated T-regulatory (Treg) and other T-lymphocyte sub-sets were evaluated. The results showed that carnosol at both doses significantly suppressed tumor growth and caused depletion of splenic and tumor-associated Treg cells. It also caused relative (vs control mouse cell values) decreases in splenocyte spontaneous/inducible production of IL-4 and IL-10 and increases in IFNγ and cell proliferation. Carnosol at either dose did not cause changes in the percentages of CD4(+) or CD8(+) lymphocytes in the spleen or in tumor-associated lymphocyte populations. The observed increases in IFNγ, decreases in IL-10 and IL-4 production, and reductions in splenic/tumor-associated Treg cell levels might be signs reflecting the potential anti-tumor activity of carnosol. Based on the findings here, it is asserted that carnosol is a likely candidate - after more complete toxicologic evaluation - for eventual use as an anti-cancer therapeutic.


Evolution of stress response in the face of unreliable environmental signals.
Most organisms live in ever-changing environments, and have to cope with a range of different conditions. Often, the set of biological traits that are needed to grow, reproduce, and survive varies between conditions. As a consequence, organisms have evolved sensory systems to detect environmental signals, and to modify the expression of biological traits in response. However, there are limits to the ability of such plastic responses to cope with changing environments. Sometimes, environmental shifts might occur suddenly, and without preceding signals, so that organisms might not have time to react. Other times, signals might be unreliable, causing organisms to prepare themselves for changes that then do not occur. Here, we focus on such unreliable signals that indicate the onset of adverse conditions. We use analytical and individual-based models to investigate the evolution of simple rules that organisms use to decide whether or not to switch to a protective state. We find evolutionary transitions towards organisms that use a combination of random switching and switching in response to the signal. We also observe that, in spatially heterogeneous environments, selection on the switching strategy depends on the composition of the population, and on population size. These results are in line with recent experiments that showed that many unicellular organisms can attain different phenotypic states in a probabilistic manner, and lead to testable predictions about how this could help organisms cope with unreliable signals.


Pneumatic AFO Powered by a Miniature Custom Compressor for Drop Foot Correction.
For active AFO applications, pneumatic remote transmission has advantages in minimizing the mass and complexity of the system due to the flexibility in placing pneumatic components and providing high back-drivability via simple valve control. However, pneumatic systems are generally tethered to large stationary air compressors, which greatly limit the practical daily usage. In this study, we implemented a wearable custom compressor that can be worn at the trunk of the body and can generate up to 1050 kPa of pressurized air to power an unilateral active AFO for dorsiflexion (DF) assistance of drop-foot patients. In order to minimize the size and weight of the custom compressor, the compression rate of the custom compressor was optimized to the rate of consumption required to power the active AFO. The finalized system can provide a maximum assistive torque of 9.8 Nm at a functional frequency of 1 Hz and the average resistive torque during free movement was 0.03 Nm. The system was tested for five hemiplegic drop-foot patients. The proposed system showed an average improvement of 12.3° of ankle peak dorsiflexion angle during the mid to late swing phase.


Deceased organ donation in India: where do we go from here?
Transplantation represents one of the best examples of the scientific achievements of medical science. However, its success has also led to some of the fiercest ethical challenges in modern medicine. Partly as a response to the uncovering of a flourishing clandestine kidney trade, the Central government promulgated the Human Organs Transplant Act (HOTA) in 1994. HOTA, along with its amendments, was a step forward in recognising concepts such as brain death. Nevertheless, there are numerous ethical challenges still to be resolved, particularly with regard to consent, incentives to donors and families, and equitable distribution of donated organs.


The latest FAD - Faecal antibody detection in cattle. Protocol and results from three UK beef farms naturally infected with gastrointestinal nematodes.
Antibodies at gastrointestinal mucosal membranes play a vital role in immunological protection against a range of pathogens, including helminths. Gastrointestinal health is central to efficient livestock production, and such infections cause significant losses. Fecal samples were taken from 114 cattle, across three beef farms, with matched blood samples taken from 22 of those animals. To achieve fecal antibody detection, a novel fecal supernatant was extracted. Fecal supernatant and serum samples were then analysed, using adapted enzyme-linked immunosorbent assay protocols, for levels of total immunoglobulin (Ig)A, IgG, IgM, and Teladorsagia circumcincta-specific IgA, IgG, IgM and IgE (in the absence of reagents for cattle-specific nematode species). Fecal nematode egg counts were conducted on all fecal samples. Assays performed successfully and showed that IgA was the predominant antibody in fecal samples, whereas IgG was predominant in serum. Total IgA in feces and serum correlated within individuals (0.581, P = 0.005), but other Ig types did not. Results support the hypothesis that the tested protocols are an effective method for the non-invasive assessment of cattle immunology. The method could be used as part of animal health assessments, although further work is required to interpret the relationship between results and levels of infection and immunity.


Identification of chemical constituents of Zanthoxylum heitzii stem bark and their insecticidal activity against the malaria mosquito Anopheles gambiae.
Zanthoxylum heitzii bark extracts have insecticidal properties and have been reported to be used against malaria in Western Africa. Previously, it has been shown that a hexane extract of the bark is toxic to adult females of the mosquito Anopheles gambiae, a malaria vector. As part of our project on the control of malaria vectors using plant extracts, the phytochemistry of Z. heitzii bark hexane extract has been investigated with the aim to identify the major components with adulticidal and larvicidal effects on An. gambiae. Z. heitzii stem bark was extracted with hexane, and the extract was fractionated to isolate major components from the bark, identified by NMR spectroscopy. Isolated compounds were tested for toxicity towards adult female An. gambiae mosquitoes and for larvicidal effects towards An. gambiae. The alkaloid dihydronitidine, the sesquiterpenoid caryophyllene oxide, the amide pellitorine and the lignan sesamin were identified as the major constituents in Z. heitzii bark. Pellitorine was toxic to both adult insects (LD50 50 ng/mg insect) and larvae (LD50 13 μg/ml). None of the other compounds were toxic to adults, but caryophyllene oxide and sesamin exhibited moderate larvicidal effects (LD50 > 150 μg/ml). A mixture of the four compounds in the same ratio as in the hexane extract showed higher toxicity (LD50 34 ng/mg insect) towards adult insects than the pure compounds. The toxicity of Z. heitzii bark hexane extract to An. gambiae is mostly due to pellitorine, although interactions between pellitorine and other, inactive constituents may enhance the activity of the extract.


Increasing carbon storage in intact African tropical forests.
The response of terrestrial vegetation to a globally changing environment is central to predictions of future levels of atmospheric carbon dioxide. The role of tropical forests is critical because they are carbon-dense and highly productive. Inventory plots across Amazonia show that old-growth forests have increased in carbon storage over recent decades, but the response of one-third of the world's tropical forests in Africa is largely unknown owing to an absence of spatially extensive observation networks. Here we report data from a ten-country network of long-term monitoring plots in African tropical forests. We find that across 79 plots (163 ha) above-ground carbon storage in live trees increased by 0.63 Mg C ha(-1) yr(-1) between 1968 and 2007 (95% confidence interval (CI), 0.22-0.94; mean interval, 1987-96). Extrapolation to unmeasured forest components (live roots, small trees, necromass) and scaling to the continent implies a total increase in carbon storage in African tropical forest trees of 0.34 Pg C yr(-1) (CI, 0.15-0.43). These reported changes in carbon storage are similar to those reported for Amazonian forests per unit area, providing evidence that increasing carbon storage in old-growth forests is a pan-tropical phenomenon. Indeed, combining all standardized inventory data from this study and from tropical America and Asia together yields a comparable figure of 0.49 Mg C ha(-1) yr(-1) (n = 156; 562 ha; CI, 0.29-0.66; mean interval, 1987-97). This indicates a carbon sink of 1.3 Pg C yr(-1) (CI, 0.8-1.6) across all tropical forests during recent decades. Taxon-specific analyses of African inventory and other data suggest that widespread changes in resource availability, such as increasing atmospheric carbon dioxide concentrations, may be the cause of the increase in carbon stocks, as some theory and models predict.


Air-drying of cells, the novel conditions for stimulated synthesis of triacylglycerol in a Green Alga, Chlorella kessleri.
Triacylglycerol is used for the production of commodities including food oils and biodiesel fuel. Microalgae can accumulate triacylglycerol under adverse environmental conditions such as nitrogen-starvation. This study explored the possibility of air-drying of green algal cells as a novel and simple protocol for enhancement of their triacylglycerol content. Chlorella kessleri cells were fixed on the surface of a glass fibre filter and then subjected to air-drying with light illumination. The dry cell weight, on a filter, increased by 2.7-fold in 96 h, the corresponding chlorophyll content ranging from 1.0 to 1.3-fold the initial one. Concomitantly, the triacylglycerol content remarkably increased to 70.3 mole% of fatty acids and 15.9% (w/w), relative to total fatty acids and dry cell weight, respectively, like in cells starved of nitrogen. Reduction of the stress of air-drying by placing the glass filter on a filter paper soaked in H2O lowered the fatty acid content of triacylglycerol to 26.4 mole% as to total fatty acids. Moreover, replacement of the H2O with culture medium further decreased the fatty acid content of triacylglycerol to 12.2 mole%. It thus seemed that severe dehydration is required for full induction of triacylglycerol synthesis, and that nutritional depletion as well as dehydration are crucial environmental factors. Meanwhile, air-drying of Chlamydomonas reinhardtii cells increased the triacylglycerol content to only 37.9 mole% of fatty acids and 4.8% (w/w), relative to total fatty acids and dry cell weight, respectively, and a marked decrease in the chlorophyll content, on a filter, of 33%. Air-drying thus has an impact on triacylglycerol synthesis in C. reinhardtii also, however, the effect is considerably limited, owing probably to instability of the photosynthetic machinery. This air-drying protocol could be useful for the development of a system for industrial production of triacylglycerol with appropriate selection of the algal species.


Alternative RNA Processing of Topoisomerase IIα in Etoposide-Resistant Human Leukemia K562 Cells: Intron Retention Results in a Novel C-Terminal Truncated 90-kDa Isoform.
DNA topoisomerase IIα (TOP2α) is a prominent target for anticancer drugs whose clinical efficacy is often limited by chemoresistance. Using antibody specific for the N-terminal of TOP2α, immunoassays indicated the existence of two TOP2α isoforms, 170 and 90 kDa, present in K562 leukemia cells and in an acquired etoposide (VP-16)-resistant clone (K/VP.5). TOP2α/90 expression was dramatically increased in etoposide-resistant K/VP.5 compared with parental K562 cells. We hypothesized that TOP2α/90 was the translation product of novel alternatively processed pre-mRNA, confirmed by 3'-rapid amplification of cDNA ends, polymerase chain reaction, and sequencing. TOP2α/90 mRNA includes retained intron 19, which harbors an in-frame stop codon, and two consensus poly(A) sites. The processed transcript is polyadenylated. TOP2α/90 mRNA encodes a 90,076-Da translation product missing the C-terminal 770 amino acids of TOP2α/170, replaced by 25 unique amino acids through translation of the exon 19/intron 19 read-through. Immunoassays, utilizing antisera raised against these unique amino acids, confirmed that TOP2α/90 is expressed in both cell types, with overexpression in K/VP.5 cells. Immunodetection of complex of enzyme-to-DNA and single-cell gel electrophoresis (Comet) assays demonstrated that K562 cells transfected with a TOP2α/90 expression plasmid exhibited reduced etoposide-mediated TOP2α-DNA covalent complexes and decreased etoposide-induced DNA damage, respectively, compared with similarly treated K562 cells transfected with empty vector. Because TOP2α/90 lacks the active site tyrosine (Tyr805) of full-length TOP2α, these results strongly suggest that TOP2α/90 exhibits dominant-negative properties. Further studies are underway to characterize the mechanism(s) by which TOP2α/90 plays a role in acquired resistance to etoposide and other TOP2α targeting agents.


Catalytic conversion of biomass to monofunctional hydrocarbons and targeted liquid-fuel classes.
It is imperative to develop more efficient processes for conversion of biomass to liquid fuels, such that the cost of these fuels would be competitive with the cost of fuels derived from petroleum. We report a catalytic approach for the conversion of carbohydrates to specific classes of hydrocarbons for use as liquid transportation fuels, based on the integration of several flow reactors operated in a cascade mode, where the effluent from the one reactor is simply fed to the next reactor. This approach can be tuned for production of branched hydrocarbons and aromatic compounds in gasoline, or longer-chain, less highly branched hydrocarbons in diesel and jet fuels. The liquid organic effluent from the first flow reactor contains monofunctional compounds, such as alcohols, ketones, carboxylic acids, and heterocycles, that can also be used to provide reactive intermediates for fine chemicals and polymers markets.


Pectosomes and chitosomes as delivery systems for metronidazole: the one-pot preparation method.
Mucoadhesive liposomes offer a potential for improved residence time of liposomal systems targeting contact with mucosal tissues, such as in buccal, oral, colon, and vaginal drug delivery. Most of the currently available methods rely on the coating of preformed liposomes by various mucoadhesive polymers. The aim of this study was to develop novel mucoadhesive system by the one-pot preparation method. The pectin- and chitosan-containing liposomes, namely pectosomes and chitosomes, were prepared by the modified solvent injection method. In order to optimize this novel delivery system, we used pectins and chitosans of both high and low degree of esterification/deacetylation (DE/DD), respectively. Sonication was applied to reduce the original vesicle size. All vesicles were characterized for their size, zeta potential, metronidazole entrapment, and stability. Both pectosomes and chitosomes were found to entrap more metronidazole than conventional plain liposomes. Preliminary data indicate that the polymer is present on the liposomal surface, embedded within inner liposomal bilayers, and entrapped inside the aqueous compartment. The next step in the evaluation of this system is the testing of its mucoadhesiveness.


Internet addiction and psychological distress among Chinese schoolchildren before and during the COVID-19 outbreak: A latent class analysis.
The present longitudinal study examined the changes in problematic internet use (problematic smartphone use, problematic social media use, and problematic gaming) and changes in COVID-19-related psychological distress (fear of COVID-19 and worry concerning COVID-19) across three time-points (before the COVID-19 outbreak, during the initial stages of the COVID-19 outbreak, and during the COVID-19 outbreak recovery period). A total of 504 Chinese schoolchildren completed measures concerning problematic internet use and psychological distress across three time-points. Latent class analysis (LCA) was used to classify participants into three groups of problematic internet use comprising Group 1 (lowest level), Group 2 (moderate level), and Group 3 (highest level). Statistical analyses showed that as problematic use of internet-related activities declined among Group 3 participants across the three time points, participants in Group 1 and Group 2 had increased problematic use of internet-related activities. Although there was no between-group difference in relation to worrying concerning COVID-19 infection, Groups 2 and 3 had significantly higher levels of fear of COVID-19 than Group 1 during the COVID-19 recovery period. Regression analysis showed that change in problematic internet use predicted fear of COVID-19 during the recovery period. The varied levels of problematic internet use among schoolchildren reflect different changing trends of additive behaviors during COVID-19 outbreak and recovery periods.


RNA Viruses of Amblyomma variegatum and Rhipicephalus microplus and Cattle Susceptibility in the French Antilles.
Ticks transmit a wide variety of pathogens including bacteria, parasites and viruses. Over the last decade, numerous novel viruses have been described in arthropods, including ticks, and their characterization has provided new insights into RNA virus diversity and evolution. However, little is known about their ability to infect vertebrates. As very few studies have described the diversity of viruses present in ticks from the Caribbean, we implemented an RNA-sequencing approach on Amblyomma variegatum and Rhipicephalus microplus ticks collected from cattle in Guadeloupe and Martinique. Among the viral communities infecting Caribbean ticks, we selected four viruses belonging to the Chuviridae, Phenuiviridae and Flaviviridae families for further characterization and designing antibody screening tests. While viral prevalence in individual tick samples revealed high infection rates, suggesting a high level of exposure of Caribbean cattle to these viruses, no seropositive animals were detected. These results suggest that the Chuviridae- and Phenuiviridae-related viruses identified in the present study are more likely tick endosymbionts, raising the question of the epidemiological significance of their occurrence in ticks, especially regarding their possible impact on tick biology and vector capacity. The characterization of these viruses might open the door to new ways of preventing and controlling tick-borne diseases.


Social support and subjective burden in caregivers of adults and older adults: A meta-analysis.
Despite the generally accepted belief that social support improves caregiver adjustment in general and subjective burden in particular, the literature shows mixed findings, and a recent review concluded that the predictive strength of caregiver social support in determining caregiver burden is less evident, due to the conceptual diversity of this determinant. The purpose of this review is to analyse the relationship of perceived and received social support with subjective burden among informal caregivers of an adult or older adult. A systematic search was carried out up to September 2017 in the following databases: MEDLINE (PubMed), CINAHL, EMBASE, PsycINFO), Scopus and ISI Proceedings, and a meta-analysis was performed with the results of the selected and included studies. Fifty-six studies were included in the meta-analysis, which provided 46 independent comparisons for perceived support and 16 for received support. Most of these studies were cross-sectional. There was a moderate, negative association of perceived social support on subjective burden (r = -0.36; CI 95% = -0.40, -0.32) and a very small, negative association of received support on subjective burden (r = -0.05; CI 95% = -0.095, -0.001). 1) perceived and received support are not redundant constructs, 2) the relationships between social support and subjective burden depend on whether the social support is measured as perceived or received, 3) the relationship of perceived social support with subjective burden has a bigger effect size than that of received social support, the relation between received support and subjective burden being clinically irrelevant, 4) perceived social support may be a good predictor of subjective burden. Our findings broadly support interventions promoting social support in caregivers to prevent or alleviate subjective burden, and specifically, to intervene on the promotion of perceived social support more than on the promotion of received social support when preventing or alleviating burden.


Time on dialysis and cancer risk after kidney transplantation.
Increasing time on dialysis is an established risk factor for certain cancer types for patients on dialysis, but the relationship between the time on dialysis and cancer risk after transplantation is unclear. We aimed to determine if the length of time on maintenance dialysis before the first kidney transplantation was associated with the risk of site-specific and overall incident cancers after transplantation. Using the Australia and New Zealand Dialysis and Transplant Registry, we assessed the association between both all cancers (except for nonmelanocytic skin cancers) and site-specific cancer incidence and the duration on dialysis before the first transplantation using adjusted Cox proportional hazards and competing risk models. Over a median follow-up of 4.4 years (interquartile range, 1.7-7.7 years), the total cumulative incidence of all cancers after the first kidney transplantation was 15.0 per 1000 patient-years. There was a linear relationship between the duration of dialysis and the risk of cancer after transplantation (Ptrend=0.02). The excess risks for lung and urinary tract cancers were highest among recipients who had been on dialysis for the longest duration before transplantation (adjusted hazard ratio [95% confidence interval], 3.32 [1.00-11.4]; P=0.05 and 2.57 [1.33-4.95]; P=0.005, respectively). Increasing time on dialysis is a significant risk factor for common solid organ cancers, such as lung cancer, and urinary tract cancers in kidney transplant recipients, irrespective of age. Strategies to improve cancer surveillance among recipients who had been on dialysis for a longer time may be warranted.


Class III malocclusion and bilateral cross-bite in an adult patient treated with miniscrew-assisted rapid palatal expander and aligners.
This case report describes the use of a miniscrew-assisted rapid palatal expander and aligners to correct bilateral cross-bite and crowding in an adult patient with a Class III skeletal pattern. A digitally designed surgical guide was three-dimensionally printed and used to accurately insert four miniscrews into the palate; these were employed to anchor a novel miniscrew-assisted rapid palatal expander appliance without any dental anchorage. Cone-beam computed tomograms before and after miniscrew-assisted rapid palatal expander treatment demonstrated the orthopedic expansion of the maxilla without dental tipping. The patient was then fitted with aligners to correct crowding and malocclusion. This case report demonstrates the successful treatment of an adult patient with a narrow maxilla and bilateral cross-bite using a nonsurgical, conservative treatment.


Breaking-Cas-interactive design of guide RNAs for CRISPR-Cas experiments for ENSEMBL genomes.
The CRISPR/Cas technology is enabling targeted genome editing in multiple organisms with unprecedented accuracy and specificity by using RNA-guided nucleases. A critical point when planning a CRISPR/Cas experiment is the design of the guide RNA (gRNA), which directs the nuclease and associated machinery to the desired genomic location. This gRNA has to fulfil the requirements of the nuclease and lack homology with other genome sites that could lead to off-target effects. Here we introduce the Breaking-Cas system for the design of gRNAs for CRISPR/Cas experiments, including those based in the Cas9 nuclease as well as others recently introduced. The server has unique features not available in other tools, including the possibility of using all eukaryotic genomes available in ENSEMBL (currently around 700), placing variable PAM sequences at 5' or 3' and setting the guide RNA length and the scores per nucleotides. It can be freely accessed at: http://bioinfogp.cnb.csic.es/tools/breakingcas, and the code is available upon request.


Endotoxin, epinephrine, glucagon, insulin and calcium ionophore A23187 modulation of pyruvate kinase activity in cultured rat hepatocytes.
Altered glucose metabolism is one of the commonly observed sequelae of sepsis and septic shock. The present investigation was undertaken to determine the role of endotoxin (ET) upon hepatocyte glucoregulation, by measuring the activity of pyruvate kinase (PK), a key glycolytic enzyme. Hepatocytes were exposed to endotoxin concentrations known to occur in vivo during sepsis, i.e., from 1 X 10(-14) to 1 X 10(-8) g/ml. The alteration of the enzyme activities after addition of epinephrine, glucagon, insulin and calcium ionophore A23187 with and without ET preincubation were also examined. ET alone decreased the PK activity by 12% at all concentrations tested. The basal inhibition of the enzyme caused by epinephrine (-48%) was partially blocked by ET preincubation above 1 X 10(-10) g/ml. There were no ET-(glucagon, calcium ionophore, insulin) interaction. These in vitro results do not support pyruvate kinase as a site of hepatic enzyme regulation defect in endotoxaemia.


Bioactivity and prostate tissue distribution of metformin in a preprostatectomy prostate cancer cohort.
Metformin has recently been shown to have potential to reduce prostate cancer risk. We conducted a randomized, double-blind, placebo-controlled trial to determine the modulating effects of metformin on tissue and systemic biomarkers of drug activity and its distribution into the prostate tissue. Twenty patients with prostate cancer scheduled to undergo prostatectomy were randomly assigned to receive either extended-release metformin or placebo for a median of 34 days before surgery. Prostatectomy and serum samples were analyzed for metformin concentrations, serum biomarkers of drug activity (prostate-specific antigen, insulin, insulin-like growth factor-1, insulin-like growth factor binding protein 3, sex hormone-binding globulin, and testosterone) and tissue biomarkers of proliferation, apoptosis, cell cycle regulation, and mTOR inhibition. For participants in the metformin arm, the prostate tissue and serum metformin concentrations ranged from 0.88 to 51.2 μg/g tissue and from not detectable to 3.6 μg/ml, respectively. There were no differences between the two groups in either the postintervention tissue biomarker expression in the prostatectomy tissue or pre to postintervention changes in serum biomarkers. We conclude that metformin distributes to human prostate tissue, suggesting that metformin could exert its effects directly on tissue targets. However, there was no difference in tissue and systemic drug effect biomarkers between the two treatment arms. Future studies with longer intervention duration and larger sample size should be considered in order to evaluate the potential of metformin for prostate cancer prevention.


Avian-like attributes of a virtual brain model of the oviraptorid theropod Conchoraptor gracilis.
An almost complete adult endoneurocranium of Conchoraptor gracilis Barsbold 1986 (Oviraptoridae; ZPAL MgD-I/95), discovered at the Hermiin Tsav locality (the Upper Cretaceous) in Mongolia, is analyzed. A virtual model of the endoneurocranial cavity was derived from CT scans and represents the most complete maniraptoran endocast to date. It displays reduced olfactory bulbs, large cerebral hemispheres in contact with the expanded cerebellum, an epiphysial projection, optic lobes displaced latero-ventrally, presumptive cerebellar folia, enlarged cerebellar auricles, and a deep medulla oblongata with a prominent ventral flexure. Contrary to Archaeopteryx, the shortened olfactory tract and cerebellum overtopping cerebral hemispheres of Conchoraptor resemble conditions in modern birds. Calculating brain mass relative to body mass indicates that Conchoraptor falls within the range of extant birds, whereas Archaeopteryx occupies a marginal position. Most of the endoneurocranial attributes, however, have a less birdlike appearance in Conchoraptor than do corresponding structures in Archaeopteryx and modern birds in which 1) postero-laterally expanded hemispheral domains broadly overlap the optic lobes, 2) the epiphysis projects to the posterior cerebrum, 3) lateral extension of the optic lobes substantially decreases a brain length-to-width ratio, 4) optic lobe and anterior hindbrain are superposed in lateral view, and 5) cerebellar and midbrain compartments are in distinct superposition. The endoneurocranial characteristics of Conchoraptor, taken together, suggest that the animal had a keen sense of vision, balance, and coordination. The data presented in this study do not allow an unambiguous assessment whether the avian-like endoneurocranial characteristics of the flightless Conchoraptor evolved convergently to those of avian theropods, or indicate a derivation of oviraptorosaurs from volant ancestors.


Diamond coating of titanium alloys.
A titanium alloy was coated with a thin layer of synthetic diamond by chemical vapor deposition methods, achieving exceptional adhesion. Scientific and technological opportunities exist for the development of diamond-coated metal alloys and for a better understanding of adhesion mechanisms of hard, brittle coatings. An indentation method of wide applicability for measuring the adhesion of such coatings is discussed.


Human Complement Bactericidal Responses to a Group A Meningococcal Conjugate Vaccine in Africans and Comparison to Responses Measured by 2 Other Group A Immunoassays.
PsA-TT (MenAfriVac) is a conjugated polysaccharide vaccine developed to eliminate group A meningococcal disease in Africa. Vaccination of African study participants with 1 dose of PsA-TT led to the production of anti-A polysaccharide antibodies and increased serum bactericidal activity measured using rabbit complement (rSBA). Bactericidal responses measured with human complement (hSBA) are presented here. Sera collected before and at 28 days and 1 year after vaccination with either PsA-TT or quadrivalent polysaccharide vaccine (PsACWY) from a random, age-distributed 360-subject subset of the Meningitis Vaccine Project study of PsA-TT in Africans aged 2-29 years were tested for hSBA. Geometric mean titer, fold-rise, and threshold analyses were compared between vaccine groups and age groups. hSBA, rSBA, and immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) results were compared and assay correlation and agreement determined. hSBA responses to PsA-TT were substantially higher than those to PsACWY at 28 days and 1 year following immunization, similar to previously reported rSBA and IgG results. The hSBA and IgG ELISA results identified differences between age groups that were not evident by rSBA. The rSBA data indicated sustained high titers 1 year after immunization, whereas hSBA GMTs at 1 year approached 4 in young children. The high level of protection following PsA-TT immunization campaigns is consistent with the strong hSBA immune responses observed here. Future implementation decisions will likely depend on immunologic data and their long-term correlation with disease and carriage prevention. Expanded immunologic and epidemiologic surveillance may improve the interpretation of differences between these immunoassays.


Uptake dynamics in the Lactose permease (LacY) membrane protein transporter.
The sugar transporter Lactose permease (LacY) of Escherichia coli has become a prototype to understand the underlying molecular details of membrane transport. Crystal structures have trapped the protein in sugar-bound states facing the periplasm, but with narrow openings unable to accommodate sugar. Therefore, the molecular details of sugar uptake remain elusive. In this work, we have used extended simulations and metadynamics sampling to explore a putative sugar-uptake pathway and associated free energy landscape. We found an entrance at helix-pair 2 and 11, which involved lipid head groups and residues Gln 241 and Gln 359. Furthermore, the protein displayed high flexibility on the periplasmic side of Phe 27, which is located at the narrowest section of the pathway. Interactions to Phe 27 enabled passage into the binding site, which was associated with a 24 ± 4 kJ/mol binding free energy in excellent agreement with an independent binding free energy calculation and experimental data. Two free energy minima corresponding to the two possible binding poses of the lactose analog β-D-galactopyranosyl-1-thio-β-D-galactopyranoside (TDG) were aligned with the crystal structure-binding pocket. This work outlines the chemical environment of a putative periplasmic sugar pathway and paves way for understanding substrate affinity and specificity in LacY.


Is inhibition of oxygen radical production of neutrophils by sympathomimetics mediated via beta-2 adrenoceptors?
Despite their beneficial effects on cardiovascular derangements in patients with severe sepsis, high doses of sympathomimetics might contribute to an impaired neutrophil function. This study was conducted to examine whether various sympathomimetics [(-)-epinephrine (EPI), dopamine (DA) and dobutamine (DOB)] differ in their potency to suppress the formation of oxygen radicals by neutrophils and whether this potency correlates with their affinity to or intrinsic activity for beta-2 adrenoceptors (beta-2 AR). Oxygen radical production of human neutrophils was induced by N-formyl-methionyl-leucyl-phenyl-alanine and detected by chemiluminescence measurements. Dose-response curves for the inhibition of chemiluminescence by sympathomimetics were measured in the absence and presence of 0.1 microM CGP 20,712 A (1-[2(3-carbamoyl-4-hydroxy phenoxy)-ethylamino]-3-[4-(1-methyl-4-trifluoromethyl-2-imidazolyl) phenoxy]-2-propanol methanesulfonate) and 0.1 microM ICI 118,551 (erythro-(+/-)-1-(7-methylindan-4-yloxy)-3 isopropylaminobutan-2-ol hydrochloride) to selectively antagonize beta-1 AR and beta-2 AR, respectively. Inhibition of chemiluminescence of neutrophils by EPI was approximately 100-fold more potent than that by DA and DOB. Only the inhibition curve by EPI exhibited two components, one at nanomolar and one at micromolar concentrations. The nanomolar component was sensitive against beta-2 AR blockade, whereas the micromolar one was insensitive against both beta AR antagonists. Dose-response curves for DA and DOB exhibited a simple hyperbolic shape at micromolar concentrations and were insensitive against both beta AR antagonists. Maximum inhibition by DA and DOB was equipotent to that by EPI. However, the EC50 for DA was much lower than its dissociation constants, KD, assayed in membrane preparations by radioligand binding, whereas the EC50 of DOB matched KD. This difference could not be explained by a different efficiency of signal transduction, which was determined in receptor-coupled adenylate cyclase activity and which only showed a slightly higher efficiency of DA (51%) than of DOB (34%). Therefore, sympathomimetics were also investigated in a cell-free system, in which chemiluminescence was generated by horseradish peroxidase with hydrogen peroxide as substrate. Surprisingly, all of the sympathomimetics suppressed chemiluminescence with micromolar concentrations. We conclude that sympathomimetics with high affinity and high intrinsic activity (EPI) inhibit neutrophil function via occupation of beta-2 AR, whereas sympathomimetics with low affinity (DA) or low intrinsic activity (DOB) may act by direct scavenging of oxygen radicals.


Probing the metallating ability of a polybasic sodium alkylmagnesiate supported by a bulky bis(amido) ligand: deprotomagnesiation reactions of nitrogen-based aromatic substrates.
Exploring the reactivity of sodium butylmagnesiate reagent [{Na(THF)6}(+){(Ph2Si(NAr*)2)Mg(Bu)(THF)}(-)] (1) supported by the bulky chelating silyl(bisamido) ligand {Ph2Si(NAr*)2}(2-) (Ar* = 2,6-iPr2-C6H3) towards N-methylbenzimidazole (bIm(Me)), pyrrole and 2,6-diisopropylaniline (NH2Ar*), this study provides new insights into the ability of this bimetallic base to facilitate direct Mg-H exchange reactions as well as to exhibit polybasicity. Thus 1 effectively promotes the deprotomagnesiation of bIm(Me) under mild reaction conditions to give the α-metallated intermediate [{Na(THF)5}2(+){(Ph2Si(NAr*)2)Mg(bIm(Me)*)}2(-)] (2) (bIm(Me)* = 2-N-methylbenzimidazolyl). Analysis of crystallographic and NMR data of 2 combined with DFT calculations show that the metallated C in the bIm(Me)* ligands possesses a significant carbenic character. Contrasting with previous studies of benzothiazole (btz), 1 does not react with an excess of bIm(Me) even under forcing refluxing conditions. Contrastingly, the amination reactions of equimolar amounts of 1 with pyrrole and 2,6-diisopropylaniline allowed the isolation of [{(Ph2Si(NAr*)(NHAr*))Mg(NC4H4)2(THF)Na(THF)2}] (3) and [{Na(THF)6}(+){(Ph2Si(NAr*)(NHAr*))Mg(NHAr*)2(THF)}(-)] (4) respectively as crystalline solids. Highlighting the ability of 1 to act as a polybasic reagent, 3 and 4 are formed as the result of the deprotonation of two molecules of the relevant amine via its butyl group and one amido arm of the silyl(bisamido) ligand. Similarly, the reactions of 1 with 3 molar equivalents of the relevant amine yielded homoleptic tris(amido) compounds [(THF)2NaMg(NC4H4)3] (5) and [{Na(THF)6}(+){Mg(NHAr*)3}(-)] (7), with the concomitant formation of bis(amine) Ph2Si(NHAr)2, as a result of the complete amination of 1 using its three basic sites. The structures in the solid state of 3 and 4 were elucidated by X-ray crystallography. Despite their similar constitution, these heteroleptic tris(amido)magnesiates adopt contrasting structures, with the former displaying a contacted ion-pair structure, where Na and Mg are connected by two bridging pyrrolyl anions, whereas the latter gives rise to a solvent-separated ion pair motif. To the best of our knowledge 3 represents the first crystallographically characterized magnesium compound containing an anionic non-substituted form of pyrrole. Noticeably, Mg interacts exclusively with the N atoms of the pyrrolyl ligands, forming strong σ-bonds, whereas Na prefers to engage with the π-systems of both NC4-rings.


Quantum correlations in optical angle-orbital angular momentum variables.
Entanglement of the properties of two separated particles constitutes a fundamental signature of quantum mechanics and is a key resource for quantum information science. We demonstrate strong Einstein, Podolsky, and Rosen correlations between the angular position and orbital angular momentum of two photons created by the nonlinear optical process of spontaneous parametric down-conversion. The discrete nature of orbital angular momentum and the continuous but periodic nature of angular position give rise to a special sort of entanglement between these two variables. The resulting correlations are found to be an order of magnitude stronger than those allowed by the uncertainty principle for independent (nonentangled) particles. Our results suggest that angular position and orbital angular momentum may find important applications in quantum information science.


Alcohol drinking exacerbates neural and behavioral pathology in the 3xTg-AD mouse model of Alzheimer's disease.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder that represents the most common cause of dementia in the United States. Although the link between alcohol use and AD has been studied, preclinical research has potential to elucidate neurobiological mechanisms that underlie this interaction. This study was designed to test the hypothesis that nondependent alcohol drinking exacerbates the onset and magnitude of AD-like neural and behavioral pathology. We first evaluated the impact of voluntary 24-h, two-bottle choice home-cage alcohol drinking on the prefrontal cortex and amygdala neuroproteome in C57BL/6J mice and found a striking association between alcohol drinking and AD-like pathology. Bioinformatics identified the AD-associated proteins MAPT (Tau), amyloid beta precursor protein (APP), and presenilin-1 (PSEN-1) as the main modulators of alcohol-sensitive protein networks that included AD-related proteins that regulate energy metabolism (ATP5D, HK1, AK1, PGAM1, CKB), cytoskeletal development (BASP1, CAP1, DPYSL2 [CRMP2], ALDOA, TUBA1A, CFL2, ACTG1), cellular/oxidative stress (HSPA5, HSPA8, ENO1, ENO2), and DNA regulation (PURA, YWHAZ). To address the impact of alcohol drinking on AD, studies were conducted using 3xTg-AD mice that express human MAPT, APP, and PSEN-1 transgenes and develop AD-like brain and behavioral pathology. 3xTg-AD and wild-type mice consumed alcohol or saccharin for 4 months. Behavioral tests were administered during a 1-month alcohol-free period. Alcohol intake induced AD-like behavioral pathologies in 3xTg-AD mice including impaired spatial memory in the Morris Water Maze, diminished sensorimotor gating as measured by prepulse inhibition, and exacerbated conditioned fear. Multiplex immunoassay conducted on brain lysates showed that alcohol drinking upregulated primary markers of AD pathology in 3xTg-AD mice: Aβ 42/40 ratio in the lateral entorhinal and prefrontal cortex and total Tau expression in the lateral entorhinal cortex, medial prefrontal cortex, and amygdala at 1-month post alcohol exposure. Immunocytochemistry showed that alcohol use upregulated expression of pTau (Ser199/Ser202) in the hippocampus, which is consistent with late-stage AD. According to the NIA-AA Research Framework, these results suggest that alcohol use is associated with Alzheimer's pathology. Results also showed that alcohol use was associated with a general reduction in Akt/mTOR signaling via several phosphoproteins (IR, IRS1, IGF1R, PTEN, ERK, mTOR, p70S6K, RPS6) in multiple brain regions including hippocampus and entorhinal cortex. Dysregulation of Akt/mTOR phosphoproteins suggests alcohol may target this pathway in AD progression. These results suggest that nondependent alcohol drinking increases the onset and magnitude of AD-like neural and behavioral pathology in 3xTg-AD mice.


DNA-cellulose: an economical, fully recyclable and highly effective chiral biomaterial for asymmetric catalysis.
The challenge in DNA-based asymmetric catalysis is to perform a reaction in the vicinity of the helix by incorporating a small-molecule catalyst anchored to the DNA in a covalent, dative, or non-covalent yet stable fashion in order to ensure high levels of enantio-discrimination. Here, we report the first generation of a DNA-based catalyst bound to a cellulose matrix. The chiral biomaterial is commercially available, trivial to use, fully recyclable and produces high levels of enantioselectivity in various Cu(II)-catalyzed asymmetric reactions including Friedel-Crafts alkylations and Michael additions. A single-pass, continuous-flow process is also reported affording fast conversions and high enantioselectivities at low catalyst loadings thus offering a new benchmark in the field of DNA-based asymmetric catalysis.


Risk behaviours for organism transmission in health care delivery-A two month unstructured observational study.
Errors in infection control practices risk patient safety. The probability for errors can increase when care practices become more multifaceted. It is therefore fundamental to track risk behaviours and potential errors in various care situations. The aim of this study was to describe care situations involving risk behaviours for organism transmission that could lead to subsequent healthcare-associated infections. Unstructured nonparticipant observations were performed at three medical wards. Healthcare personnel (n=27) were shadowed, in total 39h, on randomly selected weekdays between 7:30 am and 12 noon. Content analysis was used to inductively categorize activities into tasks and based on the character into groups. Risk behaviours for organism transmission were deductively classified into types of errors. Multiple response crosstabs procedure was used to visualize the number and proportion of errors in tasks. One-Way ANOVA with Bonferroni post Hoc test was used to determine differences among the three groups of activities. The qualitative findings gives an understanding of that risk behaviours for organism transmission goes beyond the five moments of hand hygiene and also includes the handling and placement of materials and equipment. The tasks with the highest percentage of errors were; 'personal hygiene', 'elimination' and 'dressing/wound care'. The most common types of errors in all identified tasks were; 'hand disinfection', 'glove usage', and 'placement of materials'. Significantly more errors (p<0.0001) were observed the more multifaceted (single, combined or interrupted) the activity was. The numbers and types of errors as well as the character of activities performed in care situations described in this study confirm the need to improve current infection control practices. It is fundamental that healthcare personnel practice good hand hygiene however effective preventive hygiene is complex in healthcare activities due to the multifaceted care situations, especially when activities are interrupted. A deeper understanding of infection control practices that goes beyond the sense of security by means of hand disinfection and use of gloves is needed as materials and surfaces in the care environment might be contaminated and thus pose a risk for organism transmission.


Antioxidant synergism of green tea polyphenols with alpha-tocopherol and L-ascorbic acid in SDS micelles.
The synergistic antioxidant effect of polyphenols extracted from green tea, i.e. (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin gallate (EGCG) and gallic acid (GA), with alpha-tocopherol (vitamin E) and L-ascorbic acid (vitamin C) against the peroxidation of linoleic acid has been studied in sodium dodecyl sulfate (SDS) micelles. The peroxidation was initiated thermally by a water-soluble azo initiator 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH), and the reaction kinetics were studied by monitoring the formation of linoleic acid hydroperoxides and consumption of the antioxidants. It was found that the mixture of the green tea polyphenol, vitamin E and vitamin C could act synergistically to protect lipid peroxidation. Kinetic and mechanistic studies on the antioxidation process revealed that this antioxidant synergism was due to the regeneration of vitamin E by the green tea polyphenol and the regeneration of the latter by vitamin C.


A new window into the molecular physiology of membrane proteins.
Integral membrane proteins comprise ∼25% of the human proteome. Yet, our understanding of their molecular physiology is still in its infancy. This can be attributed to two factors: the experimental challenges that arise from the difficult chemical nature of membrane proteins, and the unclear relationship between their activity and their native environment. New approaches are therefore required to address these challenges. Recent developments in mass spectrometry have shown that it is possible to study membrane proteins in a solvent-free environment and provide detailed insights into complex interactions, ligand binding and folding processes. Interestingly, not only detergent micelles but also lipid bilayer nanodiscs or bicelles can serve as a means for the gentle desolvation of membrane proteins in the gas phase. In this manner, as well as by direct addition of lipids, it is possible to study the effects of different membrane components on the structure and function of the protein components allowing us to add functional data to the least accessible part of the proteome.


Effectiveness of nutritional interventions in older adults at risk of malnutrition across different health care settings: Pooled analyses of individual participant data from nine randomized controlled trials.
Protein-energy malnutrition is a health concern among older adults. Improving nutritional status by increasing energy and protein intake likely benefits health. We therefore aimed to investigate effects of nutritional interventions in older adults (at risk of malnutrition) on change in energy intake and body weight, and explore if the intervention effect was modified by study or participants' characteristics, analysing pooled individual participant data. We searched for RCTs investigating the effect of dietary counseling, oral nutritional supplements (ONS) or both on energy intake and weight. Principle investigators of eligible studies provided individual participant data. We investigated the effect of nutritional intervention on meaningful increase in energy intake (>250 kcal/day) and meaningful weight gain (>1.0 kg). Logistic generalized estimating equations were performed and ORs with 95% CIs presented. We included data of nine studies with a total of 990 participants, aged 79.2 ± 8.2 years, 64.5% women and mean baseline BMI 23.9 ± 4.7 kg/m2. An non-significant intervention effect was observed for increase in energy intake (OR:1.59; 95% CI 0.95, 2.66) and a significant intervention effect for weight gain (OR:1.58; 95% CI 1.16, 2.17). Stratifying by type of intervention, an intervention effect on increase in energy intake was only observed for dietary counseling in combination with ONS (OR:2.28; 95% CI 1.90, 2.73). The intervention effect on increase in energy intake was greater for women, older participants, and those with lower BMI. Regarding weight gain, an intervention effect was observed for dietary counseling (OR:1.40; 95% CI 1.14, 1.73) and dietary counseling in combination with ONS (OR:2.48; 95% CI 1.92, 3.31). The intervention effect on weight gain was not influenced by participants' characteristics. Based on pooled data of older adults (at risk of malnutrition), nutritional interventions have a positive effect on energy intake and body weight. Dietary counseling combined with ONS is the most effective intervention.


Health status of adult long-term survivors of childhood cancer: a report from the Childhood Cancer Survivor Study.
Adult survivors of childhood cancer are at risk for medical and psychosocial sequelae that may adversely affect their health status. To compare the health status of adult survivors of childhood cancer and siblings and to identify factors associated with adverse outcomes. Health status was assessed in 9535 adult participants of the Childhood Cancer Survivor Study, a cohort of long-term survivors of childhood cancer who were diagnosed between 1970 and 1986. A randomly selected cohort of the survivors' siblings (n = 2916) served as a comparison group. Six health status domains were assessed: general health, mental health, functional status, activity limitations, cancer-related pain, and cancer-related anxiety/fears. The first 4 domains were assessed in the control group. Survivors were significantly more likely to report adverse general health (odds ratio [OR], 2.5; 95% confidence interval [CI], 2.1-3.0; P<.001), mental health (OR, 1.8; 95% CI, 1.6-2.1; P<.001), activity limitations (OR, 2.7; 95% CI, 2.3-3.3; P<.001), and functional impairment (OR, 5.2; 95% CI, 4.1-6.6; P<.001), compared with siblings. Forty-four percent of survivors reported at least 1 adversely affected health status domain. Sociodemographic factors associated with reporting at least 1 adverse health status domain included being female (OR, 1.4; 95% CI, 1.3-1.6; P<.001), lower level of educational attainment (OR, 2.0; 95% CI, 1.8-2.2; P<.001), and annual income less than 20 000 dollars (OR, 1.8; 95% CI, 1.6-2.1; P<.001). Relative to those survivors with childhood leukemia, an increased risk was observed for at least 1 adverse health status domain among those with bone tumors (OR, 2.1; 95% CI, 1.8-2.5; P<.001), central nervous system tumors (OR, 1.7; 95% CI, 1.5-2.0; P<.001), and sarcomas (OR, 1.2; 95% CI, 1.1-1.5; P =.01). Clinicians caring for adult survivors of childhood cancer should be aware of the substantial risk for adverse health status, especially among females, those with low educational attainment, and those with low household incomes.


Effect of a c-Met-specific, ATP-competitive small-molecule inhibitor SU11274 on human ovarian carcinoma cell growth, motility, and invasion.
Increased expression of the receptor tyrosine kinase c-Met has been shown to correlate with enhanced cell proliferation, motility, and invasion. The objectives of this study were to characterize total and activated c-Met expression in both normal and malignant human ovarian epithelial cells and to determine the effects of inhibiting the activation of c-Met on ovarian epithelial cell growth, motility, and invasion. Total c-Met was overexpressed in 82 (68%) of 119 ovarian carcinomas, as shown by immunohistochemistry. Quantitative reverse transcription-polymerase chain reaction and Western blot analyses revealed that ovarian carcinoma cell lines had higher levels of c-Met messenger RNA, total protein, and activated protein expression compared to normal ovarian epithelial cell cultures. Using a specific adenosine triphosphate-competitive small-molecule inhibitor, SU11274, activated c-Met was decreased in normal and ovarian carcinoma cell lines. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays showed that cell growth inhibition directly correlated to the level of activated c-Met detected in each cell line (r =-0.87, P = 0.012). Using modified Boyden chamber assays, ovarian carcinoma cells treated with SU11274 demonstrated significantly decreased cell motility and invasion compared to untreated cells (P = 0.003 and P < 0.001, respectively). These data indicate that c-Met is overexpressed in the majority of malignant ovarian epithelial cells both in vivo and in vitro and that decreasing activated c-Met in vitro can significantly decrease ovarian carcinoma cell growth, motility, and invasion. Developing therapies that specifically inhibit the activation of c-Met may represent a novel therapeutic modality for patients with ovarian carcinomas expressing high levels of c-Met.


Intraoperative blood flow analysis of direct revascularization procedures in patients with moyamoya disease.
Moyamoya disease is characterized by the progressive stenosis and often occlusion of the terminal internal carotid arteries, which leads to ischemic and hemorrhagic injuries. The etiology is unknown and surgical revascularization remains the mainstay treatment. We analyzed various hemodynamic factors in 292 patients with moyamoya disease, representing 496 revascularization procedures, including vessel dimension and intraoperative blood flow, using a perivascular ultrasonic flowprobe. Mean middle cerebral artery (MCA) flow rate was 4.4 ± 0.26 mL/min. After superficial temporal artery (STA)-MCA bypass surgery, flows at the microanastomosis were increased fivefold to a mean of 22.2 ± 0.8 mL/min. The MCA flows were significantly lower in the pediatric (16.2 ± 1.3 mL/min) compared with the adult (23.9 ± 1.0 mL/min; P<0.0001) population. Increased local flow rates were associated with clinical improvement. Permanent postoperative complications were low (<5%), but very high postanastomosis MCA flow was associated with postoperative stroke (31.2 ± 6.8 mL/min; P=0.045), hemorrhage (32.1 ± 10.2 mL/min; P=0.045), and transient neurologic deficits (28.6 ± 5.6 mL/min; P=0.047) compared with controls. Other flow and vessel dimension data are presented to elucidate the hemodynamic changes related to the vasculopathy and subsequent to surgical intervention.


Association of coffee, green tea, and caffeine intakes with serum concentrations of estradiol and sex hormone-binding globulin in premenopausal Japanese women.
Caffeine intake has been proposed to influence breast cancer risk. Its effect may be mediated by hormonal changes. The relationships between caffeine-containing beverages (coffee, green tea, black tea, oolong tea, and cola) and serum concentrations of estradiol and sex hormone-binding globulin were evaluated in 50 premenopausal Japanese women. Intakes of caffeine and caffeine-containing beverages were assessed by a semiquantitative food-frequency questionnaire. Blood samples were obtained from each woman on Days 11 and 22 of her menstrual cycle. High intakes of caffeinated coffee, green tea, and total caffeine were commonly correlated with increasing sex hormone-binding globulin on Days 11 and 22 of the cycle after controlling for potential confounders [Spearman correlation coefficients (r) ranged from 0.23 to 0.31]. Green tea but not caffeinated coffee intake was inversely correlated with estradiol on Day 11 of the cycle (r = -0.32, p = 0.04). Although the effect of caffeine cannot be distinguished from effects of coffee and green tea, consumption of caffeine-containing beverages appeared to favorably alter hormone levels associated with the risk of developing breast cancer.


MR1: An unconventional twist in the tail.
MR1 is a conserved molecule that binds microbial vitamin B metabolites and presents them to unconventional T cells. Lim and colleagues (2022. J. Cell Biol.https://doi.org/10.1083/jcb.202110125) uncover the role of AP2 in ensuring MR1 surface presentation, which relies on an atypical motif within the MR1 cytoplasmic tail.


Actions of snake neurotoxins on an insect nicotinic cholinergic synapse.
Here we examine the actions of six snake neurotoxins (alpha-cobratoxin from Naja naja siamensis, erabutoxin-a and b from Laticauda semifasciata; CM12 from N. haje annulifera, toxin III 4 from Notechis scutatus and a long toxin from N. haje) on nicotinic acetylcholine receptors in the cercal afferent, giant interneuron 2 synapse of the cockroach, Periplaneta americana. All toxins tested reduced responses to directly-applied ACh as well as EPSPs evoked by electrical stimulation of nerve XI with similar time courses, suggesting that their action is postsynaptic. Thus, these nicotinic receptors in a well-characterized insect synapse are sensitive to both long and short chain neurotoxins. This considerably expands the range of snake toxins that block insect nicotinic acetylcholine receptors and may enable further pharmacological distinctions between nAChR subtypes.


Microgravity studies of aqueous wet foams.
Foams and foaming pose important questions and problems for both fundamental research and practical applications. Despite the fact that foams have been extensively studied, many aspects of foam physics and chemistry still remain unclear. Experiments on foams performed under microgravity can be extended far beyond their counterpart where gravity is fully present (i.e. most experiments on Earth). They allow, in particular, observation of the wet foams obtained during the foaming process; on Earth, foams at this stage evolve too quickly due to gravity drainage and cannot be studied. This paper reviews the existing studies of foams under microgravity, which include studies in parabolic flights, in sounding rockets and in the International Space Station.


Independent origins of cultivated coconut (Cocos nucifera L.) in the old world tropics.
As a portable source of food, water, fuel, and construction materials, the coconut (Cocos nucifera L.) played a fundamental role in human migrations and the development of civilization across the humid tropics. Here we investigated the coconut's domestication history and its population genetic structure as it relates to human dispersal patterns. A sample of 1,322 coconut accessions, representing the geographical and phenotypic diversity of the species, was examined using ten microsatellite loci. Bayesian analyses reveal two highly genetically differentiated subpopulations that correspond to the Pacific and Indo-Atlantic oceanic basins. This pattern suggests independent origins of coconut cultivation in these two world regions, with persistent population structure on a global scale despite long-term human cultivation and dispersal. Pacific coconuts show additional genetic substructure corresponding to phenotypic and geographical subgroups; moreover, the traits that are most clearly associated with selection under human cultivation (dwarf habit, self-pollination, and "niu vai" fruit morphology) arose only in the Pacific. Coconuts that show evidence of genetic admixture between the Pacific and Indo-Atlantic groups occur primarily in the southwestern Indian Ocean. This pattern is consistent with human introductions of Pacific coconuts along the ancient Austronesian trade route connecting Madagascar to Southeast Asia. Admixture in coastal east Africa may also reflect later historic Arab trading along the Indian Ocean coastline. We propose two geographical origins of coconut cultivation: island Southeast Asia and southern margins of the Indian subcontinent.