Rate-limiting steps in yeast protein translation.
Deep sequencing now provides detailed snapshots of ribosome occupancy on mRNAs. We leverage these data to parameterize a computational model of translation, keeping track of every ribosome, tRNA, and mRNA molecule in a yeast cell. We determine the parameter regimes in which fast initiation or high codon bias in a transgene increases protein yield and infer the initiation rates of endogenous Saccharomyces cerevisiae genes, which vary by several orders of magnitude and correlate with 5' mRNA folding energies. Our model recapitulates the previously reported 5'-to-3' ramp of decreasing ribosome densities, although our analysis shows that this ramp is caused by rapid initiation of short genes rather than slow codons at the start of transcripts. We conclude that protein production in healthy yeast cells is typically limited by the availability of free ribosomes, whereas protein production under periods of stress can sometimes be rescued by reducing initiation or elongation rates.


Advanced approaches to characterize the human intestinal microbiota by computational meta-analysis.
We describe advanced approaches for the computational meta-analysis of a collection of independent studies, including over 1000 phylogenetic array datasets, as a means to characterize the variability of human intestinal microbiota. The human intestinal microbiota is a complex microbial community, consisting of several thousands of phylotypes, is specific for each individual, and impacts health and disease. We have developed a phylogenetic microarray, the Human Intestinal Tract Chip, to address the microbial diversity of the intestinal microbiota and used this tool to generate large datasets. It is of significant interest to use these datasets to be able to provide relations between microbial taxa, describe the extent and type of variability of the microbiota in the human gut, and establish relations between microbial taxa and their interaction with the host, intestinal location, or genotype. We present an advanced computational meta-analysis approach for studying human intestinal microbiota, outline the advantages and disadvantages of such a meta-analysis, and reflect it to analogous approaches in other fields. Finally, we illustrate the potential of this meta-analysis by identifying salient signatures of site-specific microbial communities, describe impact of genotype, and provide first examples of relevant relations between microbial taxa. We are in the process of designing and applying appropriate methods for carrying out a full meta-analysis of the present data. Beyond that, the next large challenges in future meta-analyses lie in the integration of data from several heterogeneous measurement methods such as next generation sequencing techniques, metaproteomics, or metabolomics. We have shown the feasibility of an advanced computational meta-analysis of the large datasets derived from the human intestinal microbiota.


[Association between the decline in global fertility rate and the incorporation of women to the workforce].
The global fertility rate (GFR) is defined as the mean number of children that a woman could have in a hypothetical cohort, not exposed to death during the fertile period. GFR has fallen from 3.4 to 1.9 children per women in the period 1970-2010. To explore the relationship between the fall in GFR and the incorporation of women to work in the period 1960-2011. Data from the National Statistics Institute was used. GRF was calculated using specific fertility rates for each year considering women aged 15 to 49 years. Work rates were obtained from yearly vital statistics reports. Between 1960 and 2011, GRF decreased from 5.5 to 1.9 in Chile. The first inflection occurred in 1970. In the same period, female workforce increased from 22.4 to 40.2%. To motivate the participation of female work-force without decrease the GRF allowing population replacement, it is suggested the need to create new public policies with benefits and support from the state.


Is there a need for a revised table of equivalent square fields for the determination of phantom scatter correction factors?
The use of the British Journal of Radiology (BJR) (supplement 17) tables of equivalent square fields for dose calculations is widespread. A revised version of the supplement was published recently, with a more elaborate discussion, but without changes in data given in these tables (Br. J. Radiol. suppl 25). The tables were generated for use in dose calculations, with relative beam data such as PDD, BSF, PSF, all with d(max) as the reference depth. However, the current philosophy in dose calculational methods is based on quantities defined at a reference depth, d(ref) = 10 cm, on a separation of phantom and head scatter, and on the use of the relative depth-dose or tissue-phantom ratios normalized at d(ref). By using these quantities as a starting point, problems at shallow depths related to the influence of contaminating electrons in the beam can be eliminated. Recently, a comprehensive set of phantom scatter factor data with d(ref) = 10 cm has been published for a set of square field sizes and a wide range of photon beam energies, showing that phantom scatter is a smoothly varying function of field size and quality index. It is not a priori evident that the conventional concept of equivalent squares for rectangular fields is also fully applicable for phantom scatter factors and phantom scatter related quantities at a depth of 10 cm. It was questioned whether or not new tables of equivalent square fields are needed for this purpose. In this paper, new tables have been constructed for four photon beam energies in the range of Co-60 to 25 MV (quality index from 0.572 to 0.783). The small differences between the outcome of these new tables allowed the construction of one averaged table of equivalent square fields. Phantom scatter factors were calculated for rectangular fields based on the use of the BJR table and on the use of the newly constructed tables and the differences were quantified. For Co-60 no improvements could be shown when using the new averaged table, but for beam energies of 6 to 10 MV small improvements of the order of 0.5 to 1.0% were found. For a higher beam energy of 25 MV the improvement is smaller. Deviations resulting from the BJR table are within the limits of accuracy as stated by the authors. Therefore, for clinical use, the continued use of the BJR table of equivalent squares for phantom scatter factors and phantom scatter related quantities of rectangular fields is justified, irrespective of photon beam energy.


Environmental determinants of E. coli, link with the diarrheal diseases, and indication of vulnerability criteria in tropical West Africa (Kapore, Burkina Faso).
In 2017, diarrheal diseases were responsible for 606 024 deaths in Sub-Saharan Africa. This situation is due to domestic and recreational use of polluted surface waters, deficits in hygiene, access to healthcare and drinking water, and to weak environmental and health monitoring infrastructures. Escherichia coli (E. coli) is an indicator for the enteric pathogens that cause many diarrheal diseases. The links between E. coli, diarrheal diseases and environmental parameters have not received much attention in West Africa, and few studies have assessed health risks by taking into account hazards and socio-health vulnerabilities. This case study, carried out in Burkina Faso (Bagre Reservoir), aims at filling this knowledge gap by analyzing the environmental variables that play a role in the dynamics of E. coli, cases of diarrhea, and by identifying initial vulnerability criteria. A particular focus is given to satellite-derived parameters to assess whether remote sensing can provide a useful tool to assess the health hazard. Samples of surface water were routinely collected to measure E. coli, enterococci and suspended particulate matter (SPM) at a monitoring point (Kapore) during one year. In addition, satellite data were used to estimate precipitation, water level, Normalized Difference Vegetation Index (NDVI) and SPM. Monthly epidemiological data for cases of diarrhea from three health centers were also collected and compared with microbiological and environmental data. Finally, semi-structured interviews were carried out to document the use of water resources, contact with elements of the hydrographic network, health behavior and condition, and water and health policy and prevention, in order to identify the initial vulnerability criteria. A positive correlation between E. coli and enterococci in surface waters was found indicating that E. coli is an acceptable indicator of fecal contamination in this region. E. coli and diarrheal diseases were strongly correlated with monsoonal precipitation, in situ SPM, and Near Infra-Red (NIR) band between March and November. Partial least squares regression showed that E. coli concentration was strongly associated with precipitation, Sentinel-2 reflectance in the NIR and SPM, and that the cases of diarrhea were strongly associated with precipitation, NIR, E. coli, SPM, and to a lesser extent with NDVI. Moreover, E. coli dynamics were reproduced using satellite data alone, particularly from February to mid-December (R2 = 0.60) as were cases of diarrhea throughout the year (R2 = 0.76). This implies that satellite data could provide an important contribution to water quality monitoring. Finally, the vulnerability of the population was found to increase during the rainy season due to reduced accessibility to healthcare and drinking water sources and increased use of water of poor quality. During this period, surface water is used because it is close to habitations, easy to use and free from monetary or political constraints. This vulnerability is aggravated by marginality and particularly affects the Fulani, whose concessions are often close to surface water (river, lake) and far from health centers.


Natranaerobius thermophilus gen. nov., sp. nov., a halophilic, alkalithermophilic bacterium from soda lakes of the Wadi An Natrun, Egypt, and proposal of Natranaerobiaceae fam. nov. and Natranaerobiales ord. nov.
Novel halophilic, alkalithermophilic, Gram-type-positive bacterial strains were isolated from sediment of alkaline, hypersaline lakes of the Wadi An Natrun, Egypt. Cells of strain JW/NM-WN-LFT were rod-shaped, non-spore-forming and non-motile. Strain JW/NM-WN-LFT grew (at pH55 degrees C 9.5) between 35 and 56 degrees C, with an optimum at 53 degrees C. The pH55 degrees C range for growth was 8.3-10.6, with an optimum at pH55 degrees C 9.5 and no growth at pH55 degrees C 8.2 or below, or at pH55 degrees C 10.8 or above. At the optimum pH and temperature, the strain grew in the Na+ range of 3.1-4.9 M (1.5-3.3 M added NaCl) and optimally between 3.3 and 3.9 M Na+ (1.7-2.3 M added NaCl). Strain JW/NM-WN-LFT utilized fructose, cellobiose, ribose, trehalose, trimethylamine, pyruvate, Casamino acids, acetate, xylose and peptone as carbon and energy sources. Fumarate (20 mM), S2O3(2-) (20 mM), NO3- (20 mM) and iron(III) citrate (20 mM) were utilized as electron acceptors. During growth on sucrose, the isolate produced acetate and formate as major fermentation products. Main cellular fatty acids were iso-branched 15:0, i17:0 dimethylacetal and 16:0 dimethylacetal. The G+C content of genomic DNA was 40.4 mol% (HPLC). On the basis of genotypic and phenotypic characteristics, it is proposed that strain JW/NM-WN-LFT represents a novel genus and species, Natranaerobius thermophilus gen. nov., sp. nov. The type strain is JW/NM-WN-LFT (=DSM 18059T=ATCC BAA-1301T). Based on 16S rRNA gene sequence analysis, the strain forms a novel lineage within the class 'Clostridia' and clusters with uncultivated bacteria and unidentified strains retrieved from alkaline, hypersaline environments. The phylogenetic data suggest that the lineage represents a novel family, Natranaerobiaceae fam. nov., and order, Natranaerobiales ord. nov.


Disseminated necrotizing panniculitis and pancreatic nodular hyperplasia in a dog.
An 8-year-old female Poodle had severe necrotizing skin disease characterized by numerous large red-purple, nonpruritic lesions. The larger lesions had soft, intensely discolored centers that contained sterile viscous blood-tinged material. Biopsies of the lesions revealed necrotizing panniculitis (subcutaneous fat necrosis). The dog did not respond to intensive therapy and euthanasia was performed. Necropsy findings included diffuse fat necrosis and pancreatic nodular hyperplasia. A pathoetiologic relation between pancreatic disease and diffuse fat necrosis in this dog was suggested.


Boar taint detection using parasitoid biosensors.
The off-flavor boar taint associated with the substances skatole, androstenone, and possibly indole represents a significant problem in the pig husbandry industry. Boar taint may occur in meat from uncastrated sexually mature male pigs; consumers commonly show a strong aversion to tainted meat. Consequently, there is a need for rapid methods to sort out and remove tainted carcasses at the slaughterline. We tested the ability of wasps, Microplitis croceipes to perceive and learn the 3 boar taint compounds both individually and in combination using classical conditioning paradigms. We also established the effectiveness and reliability of boar taint odor detection when wasps were used as biosensors in a contained system called the "wasp hound" using a cohort of trained wasps. We found that the wasps are able to successfully learn indole, skatole and to also detect them when presented a 1:1:1 mixture of all 3 compounds. This was shown for both a single hand-manipulated wasp bioassay and when using the "wasp hound" detector device. In contrast, the wasps showed a weak conditioned response to androstenone at the concentration tested. The estimated gas phase concentrations that the wasps perceived during training were in the range of 10 ± 0.4 pg/s for skatole and indole, and 2 ± 0.5 pg/s for androstenone. We conclude that use of these wasps as biosensors presents a promising method for boar taint detection and discuss future training paradigms that may improve their responses to compounds such as androstenone. Practical Application: The development of a perceptive, inexpensive, and reliable means of detecting boar taint before the product is presented to sensitive consumers.


Zebrafish knockout of Down syndrome gene, DYRK1A, shows social impairments relevant to autism.
DYRK1A maps to the Down syndrome critical region at 21q22. Mutations in this kinase-encoding gene have been reported to cause microcephaly associated with either intellectual disability or autism in humans. Intellectual disability accompanied by microcephaly was recapitulated in a murine model by overexpressing Dyrk1a which mimicked Down syndrome phenotypes. However, given embryonic lethality in homozygous knockout (KO) mice, no murine model studies could present sufficient evidence to link Dyrk1a dysfunction with autism. To understand the molecular mechanisms underlying microcephaly and autism spectrum disorders (ASD), we established an in vivo dyrk1aa KO model using zebrafish. We identified a patient with a mutation in the DYRK1A gene using microarray analysis. Circumventing the barrier of murine model studies, we generated a dyrk1aa KO zebrafish using transcription activator-like effector nuclease (TALEN)-mediated genome editing. For social behavioral tests, we have established a social interaction test, shoaling assay, and group behavior assay. For molecular analysis, we examined the neuronal activity in specific brain regions of dyrk1aa KO zebrafish through in situ hybridization with various probes including c-fos and crh which are the molecular markers for stress response. Microarray detected an intragenic microdeletion of DYRK1A in an individual with microcephaly and autism. From behavioral tests of social interaction and group behavior, dyrk1aa KO zebrafish exhibited social impairments that reproduce human phenotypes of autism in a vertebrate animal model. Social impairment in dyrk1aa KO zebrafish was further confirmed by molecular analysis of c-fos and crh expression. Transcriptional expression of c-fos and crh was lower than that of wild type fish in specific hypothalamic regions, suggesting that KO fish brains are less activated by social context. In this study, we established a zebrafish model to validate a candidate gene for autism in a vertebrate animal. These results illustrate the functional deficiency of DYRK1A as an underlying disease mechanism for autism. We also propose simple social behavioral assays as a tool for the broader study of autism candidate genes.


Prism adaptation treatment for upper-limb complex regional pain syndrome: a double-blind randomized controlled trial.
Initial evidence suggested that people with complex regional pain syndrome (CRPS) have reduced attention to the affected side of their body and the surrounding space, which might be related to pain and other clinical symptoms. Three previous unblinded, uncontrolled studies showed pain relief after treatment with prism adaptation, an intervention that has been used to counter lateralised attention bias in brain-lesioned patients. To provide a robust test of its effectiveness for CRPS, we conducted a double-blind randomized controlled trial of prism adaptation for unilateral upper-limb CRPS-I. Forty-nine eligible adults with CRPS were randomized to undergo 2 weeks of twice-daily home-based prism adaptation treatment (n = 23) or sham treatment (n = 26). Outcomes were assessed in person 4 weeks before and immediately before treatment, and immediately after and 4 weeks after treatment. Long-term postal follow-ups were conducted 3 and 6 months after treatment. We examined the effects of prism adaptation vs sham treatment on current pain intensity and the CRPS symptom severity score (primary outcomes), as well as sensory, motor, and autonomic functions, self-reported psychological functioning, and experimentally tested neuropsychological functions (secondary outcomes). We found no evidence that primary or secondary outcomes differed between the prism adaptation and sham treatment groups when tested at either time point after treatment. Overall, CRPS severity significantly decreased over time for both groups, but we found no benefits of prism adaptation beyond sham treatment. Our findings do not support the efficacy of prism adaptation treatment for relieving upper-limb CRPS-I. This trial was prospectively registered (ISRCTN46828292).


[Psychosis during and after disulfiram use].
A 47-year-old man was registered for treatment of his alcohol abuse problem. Disulfiram was administered as part of the treatment, the intake of which was supervised by his wife. During the treatment with disulfiram and two weeks after the withdrawal of this treatment he developed a psychosis. He stated that he had not taken alcohol. During the second psychotic period he was successfully treated with antipsychotic drugs. Afterwards it was discovered that his family history was positive for schizophrenia; it is therefore possible that the patient was more vulnerable for a psychosis due to disulfiram use. Although a psychotic reaction during and after disulfiram therapy is not common, it should nonetheless be taken into consideration when prescribing the drug.


Is primary prevention of childhood obesity by education at 13-month immunisations feasible and acceptable? Results from a general practice based pilot study.
Abstract Prevalence of childhood overweight and obesity remains high in Ireland. In this study an intervention conducted within primary care was evaluated. This involved a structured discussion with parents at the 13 month immunisations with their general practitioner (GP), including measuring weight of the toddler and parental education regarding healthy nutrition and physical activity for their toddler. There was a telephone follow-up interview with parents three months later assessing change in toddler diet/lifestyle. Endpoints assessed included parents' reports of specific lifestyle parameters with regard to the toddler and parental assessment of the usefulness of the intervention. 39 toddlers were studied. Most lifestyle parameters had improved at follow up. Reported fruit and vegetable intake of more than 4 portions per day increased from 20.5% of toddlers at baseline 28.6% at follow up. The number of toddlers abstaining from unhealthy snacks increased from 15.4% to 21.4%. Television watching of more than 2 hours daily decreased from 12.8% to 0%. Supervised exercise of more than thirty minutes per day increased from 69.2% to 89.3%. The majority of parents reported at follow up that they found the intervention acceptable (100%, n = 28) and useful (79%, n = 22).


Supply Considerations for Scaling Up Clean Cooking Fuels for Household Energy in Low- and Middle-Income Countries.
Promoting access to clean household cooking energy is an important subject for policy making in low- and middle-income countries, in light of urgent and global efforts to achieve universal energy access by 2030 (Sustainable Development Goal 7). In 2014, the World Health Organization issued "Guidelines for Indoor Air Quality: Household Fuel Combustion", which recommended a shift to cleaner fuels rather than promotion of technologies that more efficiently combust solid fuels. This study fills an important gap in the literature on transitions to household use of clean cooking energy by reviewing supply chain considerations for clean fuel options in low- and middle-income countries. For the purpose of this study, we consider electricity, liquefied petroleum gas (LPG), alcohol fuels, biogas, and compressed biomass pellets burned in high performing gasifier stoves to be clean fuel options. Each of the clean fuels reviewed in this study, as well as the supply of electricity, presents both constraints and opportunities for enhanced production, supply, delivery, and long-term sustainability and scalability in resource-poor settings. These options are reviewed and discussed together with policy and regulatory considerations to help in making these fuel and energy choices available and affordable. Our hope is that researchers, government officials and policy makers, and development agencies and investors will be aided by our comparative analysis of these clean household energy choices.


Central alpha-1-adrenergic control of vasopressin secretion in newborn lambs.
The hypothesis that central alpha-1 adrenoceptors are inhibitory to the hypotension-induced secretion of vasopressin was tested by subjecting lambs that were instrumented for a long term to varying degrees of hypotension after intracerebroventricular injections of prazosin or placebo. Eight lambs in the 1st wk of life treated with intracerebroventricular injections of placebo had their mean arterial blood pressures decreased 14 and 21% by i.v. infusion of nitroprusside. Arginine vasopressin levels rose to 7.3 +/- 2.4 pmol/L only with the greater degree of hypotension. When the lambs were treated with intracerebroventricular injections of 1 micrograms/kg of prazosin, the blood pressures were decreased 13 and 23%, and the vasopressin levels were 15.4 +/- 16.6 and 27.5 +/- 20.3 pmol/L, respectively. A relationship was shown between the degree of hypotension and the plasma arginine vasopressin levels with both the placebo and prazosin, the slope being much steeper for the prazosin treatment (-1.11) than for the placebo treatment (-0.31). Plasma renin activity was increased a similar amount in both groups, and there was no change in plasma cortisol levels. We conclude that alpha-1 adrenoceptors in the brain are inhibitory to the secretion of arginine vasopressin. These results differ from observations in adult rats and dogs and may be accounted for by developmental or species differences.


Oral health of preschool children in Dhanbad District, Jharkhand: A gander into the maternal behavior and practices.
Mothers have an important role in taking decisions about their children's oral health. The study assessed the echelon of oral health behavior and practices of the mothers in relation to their preschool-aged children, which could possibly influence their oral health. A cross-sectional study was carried out among 312 mothers, selected randomly from eight blocks of Dhanbad District, Jharkhand State, by administering a structured questionnaire on oral health behavior and practices regarding oral health. The overall data indicated that the practices of mothers about oral health of their preschool children increased as their age increased, which was however, not statistically significant (P > 0.47). The mothers who had less than higher secondary school education and belonging to lower socioeconomic groups had poorer oral health practices and was found to be statistically significant (P < 0.002). The mothers ought to inculcate positive practices and perceptions about preventive oral health for themselves first, to be good role models for their children.


Dissecting aneurysms of coronary arteries.
A case of dissecting aneurysms of coronary arteries caused by cardiac surgery was found. Review discloses that most coronary arteries arise spontaneously, occur more often in women than men, frequently in the postpartum period, and are characterized by the sudden onset of acute chest pain in a previously well middle aged patient who abruptly collapses and dies. The anterior descending branch of the left coronary artery is most frequently affected, the dissection usually involves the first portion of that single vessel, and the lesion appears amenable to bypass venous grafting. While the cause is unknown, the frequency of pregnancy, the loosening effect of that condition on connective tissue, and the occasional finding of medial degeneration of the arterial wall suggest cystic medial necrosis as an important etiologic consideration. This case report recommends gentle handling of coronary arteries in surgical procedures.


Sharpness recognition based on synergy between bio-inspired nociceptors and tactile mechanoreceptors.
Touch and pain sensations are complementary aspects of daily life that convey crucial information about the environment while also providing protection to our body. Technological advancements in prosthesis design and control mechanisms assist amputees to regain lost function but often they have no meaningful tactile feedback or perception. In the present study, we propose a bio-inspired tactile system with a population of 23 digital afferents: 12 RA-I, 6 SA-I, and 5 nociceptors. Indeed, the functional concept of the nociceptor is implemented on the FPGA for the first time. One of the main features of biological tactile afferents is that their distal axon branches in the skin, creating complex receptive fields. Given these physiological observations, the bio-inspired afferents are randomly connected to the several neighboring mechanoreceptors with different weights to form their own receptive field. To test the performance of the proposed neuromorphic chip in sharpness detection, a robotic system with three-degree of freedom equipped with the tactile sensor indents the 3D-printed objects. Spike responses of the biomimetic afferents are then collected for analysis by rate and temporal coding algorithms. In this way, the impact of the innervation mechanism and collaboration of afferents and nociceptors on sharpness recognition are investigated. Our findings suggest that the synergy between sensory afferents and nociceptors conveys more information about tactile stimuli which in turn leads to the robustness of the proposed neuromorphic system against damage to the taxels or afferents. Moreover, it is illustrated that spiking activity of the biomimetic nociceptors is amplified as the sharpness increases which can be considered as a feedback mechanism for prosthesis protection. This neuromorphic approach advances the development of prosthesis to include the sensory feedback and to distinguish innocuous (non-painful) and noxious (painful) stimuli.


Potential anticancer heterometallic Fe-Au and Fe-Pd agents: initial mechanistic insights.
A series of gold(III) and palladium(II) heterometallic complexes with new iminophosphorane ligands derived from ferrocenylphosphanes [{Cp-P(Ph2)═N-Ph}2Fe] (1), [{Cp-P(Ph2)═N-CH2-2-NC5H4}2Fe] (2), and [{Cp-P(Ph2)═N-CH2-2-NC5H4}Fe(Cp)] (3) have been synthesized and structurally characterized. Ligands 2 and 3 afford stable coordination complexes [AuCl2(3)]ClO4, [{AuCl2}2(2)](ClO4)2, [PdCl2(3)], and [{PdCl2}2(2)]. The complexes have been evaluated for their antiproliferative properties in human ovarian cancer cells sensitive and resistant to cisplatin (A2780S/R), in human breast cancer cells (MCF7) and in a nontumorigenic human embryonic kidney cell line (HEK-293T). The highly cytotoxic trimetallic derivatives M2Fe (M = Au, Pd) are more cytotoxic to cancer cells than their corresponding monometallic fragments. Moreover, these complexes were significantly more cytotoxic than cisplatin in the resistant A2780R and the MCF7 cell lines. Studies of the interactions of the trimetallic compounds with DNA and the zinc-finger protein PARP-1 indicate that they exert anticancer effects in vitro based on different mechanisms of actions with respect to cisplatin.


Efficient association mapping from k-mers-An application in finding sex-specific sequences.
Genome wide association studies (GWAS) attempt to map genotypes to phenotypes in organisms. This is typically performed by genotyping individuals using microarray or by aligning whole genome sequencing reads to a reference genome. Both approaches require knowledge of a reference genome which hinders their application to organisms with no or incomplete reference genomes. This caveat can be removed by using alignment-free association mapping methods based on k-mers from sequencing reads. Here we present an improved implementation of an alignment free association mapping method. The new implementation is faster and includes additional features to make it more flexible than the original implementation. We have tested our implementation on an E. Coli ampicillin resistance dataset and observe improvement in execution time over the original implementation while maintaining accuracy in results. We also demonstrate that the method can be applied to find sex specific sequences.


Cervical cancer awareness and preventive practices: a challenge for female urban slum dwellers in Lagos, Nigeria.
Cervical cancer is the commonest gynaecological cancer in Nigeria and women of low socio-economic status are at high risk of this condition. A study was conducted on the awareness of cervical cancer, attitude towards the disease and screening practice of women residing in two urban slums of Lagos, Nigeria. It also determined the prevalence of major risk factors for cervical cancer among the women. Multistage sampling was used to select 240 women who were interviewed with a structured questionnaire and data collected was analyzed with Epi-info version 3.5.1 statistical software. Only 10 (4.2%) women in this study were aware of cervical cancer and none of them believed they were at risk of developing the disease. Most (73.3%) were willing to undergo a cervical cancer screening test. Age, education and previous history of vaginal examination were positively associated with willingness to undergo screening (p < 0.05). The respondents had a high prevalence of major risk factors for cervical cancer such as early age at sexual debut, multiple sexual partners and male partner with other female partners. Efforts need to be intensified to increase awareness of this condition and to promote low-cost cervical cancer screening among this underserved population.


Intraspecific diversity within Ganoderma lucidum in the production of laccase and Mn-oxidizing peroxidases during plant residues fermentation.
Comparison of the potential for laccase and Mn-oxidizing peroxidases synthesis by ten strains of Ganoderma lucidum, originating from different worldwide areas, during solid-state fermentation of selected plant raw materials was the aim of this study. The great intraspecific variability in the production of analyzed enzymes as well as the dependence of the enzyme activity on plant raw materials were reported. The strain HAI 957 was the best laccase producer in the presence of corn stem, as a unique carbon source (129.46 U/L). The highest level of Mn-dependent peroxidase activity was noted after wheat straw fermentation by G. lucidum HAI 246 (78.64 U/L), while the maximal versatile peroxidase production (59.72 U/L) was observed in strain HAI 957 in the medium with oak sawdust.


The proneural factors Ascl1a and Ascl1b contribute to the terminal differentiation of dopaminergic GABAergic dual transmitter neurons in zebrafish.
The proneural factor Ascl1 is involved in several steps of neurogenesis, from neural progenitor maintenance to initiation of terminal differentiation and neuronal subtype specification. In neural progenitor cells, Ascl1 initiates the cell-cycle exit of progenitors, and contributes to their differentiation into mainly GABAergic neurons. Several catecholaminergic neuron groups in the forebrain of zebrafish use GABA as co-transmitter, but a potential role of the two paralogues Ascl1a and Ascl1b in their neurogenesis is not understood. Here, we show that ascl1a, ascl1b double mutant embryos develop a significantly reduced number of neurons in all GABAergic and catecholaminergic dual transmitter neuron anatomical clusters in the fore- and hindbrain, while glutamatergic catecholaminergic clusters develop normally. However, none of the affected catecholaminergic cell clusters are lost completely, suggesting an impairment in progenitor pools, or a requirement of Ascl1a/b for differentiation of a subset of neurons in each cluster. Early progenitors which are dlx2a+, fezf2 + or emx2 + are not reduced whereas late progenitors and differentiating neurons marked by the expression of dlx5a, isl1 and arxa are severely reduced in ascl1a, ascl1b double mutant embryos. This suggests that Ascl1a and Ascl1b play only a minor or no role in the maintenance of their progenitor pools, but rather contribute to the initiation of terminal differentiation of GABAergic catecholaminergic neurons.


Vector for IS element entrapment and functional characterization based on turning on expression of distal promoterless genes.
We constructed and characterized a novel trap vector for rapid isolation of insertion sequences. The strategy used for the isolation of IS elements is based on the ability of many IS elements to turn on the expression of otherwise silent genes distal to some sites of insertion. The simple transposition of an IS element can sometimes cause the constitutive expression of promoterless antibiotic resistance genes resulting in selectable phenotypes. The trap vector pAW1326 is based on a pBR322 replicon, it carries ampicillin and streptomycin resistance genes, and also silenced genes that confer chloramphenicol and kanamycin resistance once activated. The trap vector pAW1326 proved to be efficient and 85 percent of all isolated mutations were insertions. The majority of IS elements resident in the studied Escherichia coli strains tested became trapped, namely IS2, IS3, IS5, IS150, IS186 and Tn1000. We also encountered an insertion sequence, called IS10L/R-2, which is a hybrid of the two IS variants IS10L and IS10R. IS10L/R-2 is absent from most E. coli strains, but it is detectable in some strains such as JM109 which had been submitted to Tn10 mutagenesis. The distribution of the insertion sequences within the trap region was not random. Rather, the integration of chromosomal mobile genetic elements into the offered target sequence occurred in element-specific clusters. This is explained both by the target specificity and by the specific requirements for the activation of gene transcription by the DNA rearrangement. The employed trap vector pAW1326 proved to be useful for the isolation of mobile genetic elements, for a demonstration of their transposition activity as well as for the further characterization of some of the functional parameters of transposition.


[Enhancement of toxin production and sporogenesis in Cl. tetani 471 cultures under the influence of endogenous metabolites].
A study was made of the dynamics of the population quantity, the delicate morphology of the bacterial cells and the toxin formation in the Cl. tetani 471 cultures grown in fluid nutrient medium with caseine hydrolysate in the industrial 100-litre reactors and under laboratory conditions. Endogenous metabolites contained in the filtrates of the periodic Cl. tetani 471 cultures which reached the phase of accelerated death were capable of enhancement of the toxin formation when introduced in the amount of up to 1 : 100 at the beginning of the exponential growth of the same cultures in the reactors. The growth of the some oligosporogenic strain in the filtrates of the periodic cultures obtained at the beginning of the stationary phase or of the accelerated death phase was characterized by a slower increase in the vegetative cell count, by enhanced spore-formation and by the second, after the initial, increase in the concentration of the microbes partially connected with the spore vegetation.


Charge distribution as a tool to investigate structural details. III. Extension to description in terms of anion-centred polyhedra.
The charge distribution (CHARDI) method is a self-consistent generalization of Pauling's concept of bond strength which does not make use of empirical parameters but exploits the experimental geometry of the coordination polyhedra building a crystal structure. In the two previous articles of this series [Nespolo et al. (1999). Acta Cryst. B55, 902-916; Nespolo et al. (2001). Acta Cryst. B57, 652-664], we have presented the features and advantages of this approach and its extension to distorted and heterovalent polyhedra and to hydrogen bonds. In this third article we generalize CHARDI to structures based on anion-centred polyhedra, which have drawn attention in recent years, and we show that computations based on both descriptions can be useful to obtain a deeper insight into the structural details, in particular for mixed-valence compounds where CHARDI is able to give precise indications on the statistical distribution of atoms with different oxidation number. A graph-theoretical description of the structures rationalizes and gives further support to the conclusions obtained via the CHARDI approach.


Dramatic improvement of left ventricular function after cytotoxic therapy in lupus patients with acute cardiomyopathy: report of 6 cases.
Although lupus cardiomyopathy is thought to be clinically uncommon, we encountered 6 patients with systemic lupus erythematosus (SLE) over a 10 year period who had severe left ventricular dysfunction and showed remarkable improvement in their cardiac function after cytotoxic therapy. All patients met the American College of Rheumatology criteria for classification of SLE and presented with signs of severe biventricular failure relatively early in their disease. Concurrent manifestations of SLE at the time of cardiomyopathy included rash, arthritis, myalgias, pleuritis, pericarditis, and nephritis. Four of the 6 patients were taking prednisone 20 mg/day at the time heart failure developed. In all cases the CPK were normal. Evaluation of cardiac function by echocardiogram and/or radionuclide gated blood pool scintigraphy revealed a severe depression of ventricular function with initial left ventricular ejection fraction (LVEF) ranging from 11 to 34% (mean 19%). Within 6 months of initiation of cytotoxic treatment all patients showed a dramatic response: the post-treatment LVEF ranged from 25 to 55%. This series of patients suggests that cardiomyopathy may be a more common complication of SLE than previously reported. Cardiomyopathy occurs relatively early in the course of SLE, may lead to severe cardiac dysfunction despite corticosteroid therapy, and appears to be responsive to cytotoxic therapy.


Comparison of the effects of papain and vitamin A on cartilage. II. The effects on organ cultures of embryonic skeletal tissue.
The effects of papain protease and of vitamin A on explanted limb bone rudiments from 7- and 13-day chick embryos and fetal mice have been studied and compared. The incubation of cartilaginous rudiments from 7-day chick embryos in a solution containing papain and cysteine resulted in complete loss of the metachromasia of the cartilage matrix within 1 hour; explants treated in this fashion recovered normal metachromatic staining properties when grown in normal medium for 4 days. The incubation of 7-day chick cartilage rudiments in a solution containing papain without cysteine resulted in partial loss of metachromasia from cartilage within 1 hour; the addition of vitamin A to the solution did not enhance the effect of papain during this period. The addition of papain to the culture medium in which 7-day chick embryo cartilage rudiments were grown resulted in uniform loss of the metachromasia of the cartilage matrix; similar explants grown in the presence of excess vitamin A also showed loss of the metachromasia of cartilage, but certain regions of the cartilage were affected earlier and more severely than others. Changes in cartilage cells, including loss of glycogen, occurred when the rudiment was grown in medium containing excess vitamin A, but not when it was grown in the presence of papain. Bone rudiments from 13-day chick embryos showed changes in cartilage similar to those seen in 7-day chick embryo rudiments when grown in the presence of papain or of excess vitamin A; the existing bone was not affected under these conditions. When grown in the presence of papain or excess vitamin A, the cartilage of late fetal mouse bone underwent changes similar to those already described in chick embryo rudiments. In contrast to the chick embryo rudiments, those from the fetal mouse showed rapid resorption of bone when grown in the presence of excess vitamin A. Papain had no effect on bone from either source. The changes seen in cartilage of explants grown in the presence of vitamin A and papain together were greater than those seen with either agent alone. The changes seen in fetal mouse bone grown in the presence of vitamin A were not enhanced by the additional presence of papain. On the basis of these observations, it is suggested that the changes in cartilage seen in experimental hypervitaminosis A may be the result of activation of a proteolytic enzyme or enzymes with properties similar to papain.


A Subset of Latency-Reversing Agents Expose HIV-Infected Resting CD4+ T-Cells to Recognition by Cytotoxic T-Lymphocytes.
Resting CD4+ T-cells harboring inducible HIV proviruses are a critical reservoir in antiretroviral therapy (ART)-treated subjects. These cells express little to no viral protein, and thus neither die by viral cytopathic effects, nor are efficiently cleared by immune effectors. Elimination of this reservoir is theoretically possible by combining latency-reversing agents (LRAs) with immune effectors, such as CD8+ T-cells. However, the relative efficacy of different LRAs in sensitizing latently-infected cells for recognition by HIV-specific CD8+ T-cells has not been determined. To address this, we developed an assay that utilizes HIV-specific CD8+ T-cell clones as biosensors for HIV antigen expression. By testing multiple CD8+ T-cell clones against a primary cell model of HIV latency, we identified several single agents that primed latently-infected cells for CD8+ T-cell recognition, including IL-2, IL-15, two IL-15 superagonists (IL-15SA and ALT-803), prostratin, and the TLR-2 ligand Pam3CSK4. In contrast, we did not observe CD8+ T-cell recognition of target cells following treatment with histone deacetylase inhibitors or with hexamethylene bisacetamide (HMBA). In further experiments we demonstrate that a clinically achievable concentration of the IL-15 superagonist 'ALT-803', an agent presently in clinical trials for solid and hematological tumors, primes the natural ex vivo reservoir for CD8+ T-cell recognition. Thus, our results establish a novel experimental approach for comparative evaluation of LRAs, and highlight ALT-803 as an LRA with the potential to synergize with CD8+ T-cells in HIV eradication strategies.


Region of hepatitis B virus DNA with a homology to the yeast ARS replication enhancer.
A computer analysis of the primary sequence of hepatitis B virus (HBV) subtype ayw DNA, cloned within the pVG2 recombinant plasmid, which raises its stability in Saccharomyces cerevisiae transformants, was performed. This revealed that the structure of the HBV DNA has: two bends in the termination regions of the HBs and HBc genes, and multiple sequences with a high degree of homology to the ARS (autonomously replicating sequence) core consensus in this region of the HBs gene. DNA fragments from the HBs region (330 bp) and from the HBc region (378 bp) have an abnormal electrophoretic mobility in 8% polyacrylamide gels. The similarity of the structural motifs in the stop-region of HBs gene with the B-domain of the S. cerevisiae ARS element is discussed.


Expression of an Extracellular Protein (SMU.63) Is Regulated by SprV in Streptococcus mutans.
In Streptococcus mutans, SprV (SMU.2137) is a pleiotropic regulator that differentially regulates genes related to competence, mutacin production, biofilm formation, and the stress tolerance response, along with some other pathways. In this study, we established a link between SprV and an ∼67-kDa protein in the culture supernatant of strain UA159 that was later confirmed as SMU.63 by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. We discovered that SprV downregulates the transcription and translation of SMU.63. We found that the seven amino acids from the C-terminal region of SprV were also crucial for the expression of SMU.63. Deletion of smu.63 led to increased sucrose-independent biofilm formation and competence. The sprV deletion also increased biofilm formation although this could be partially attributed to the downregulation of smu.63 In an smu.63 sprV double mutant, a synergistic effect was observed in biofilm formation in contrast to effects on competence development. We found that low or excess magnesium ion repressed sprV transcription that, in turn, affected the expression of smu.63 As expected, a magnesium ion-dependent effect of competence and biofilm formation was observed in the UA159 strain. We also replicated the results of SMU.63 expression and competence in S. mutans GS5 that encodes both SprV and SMU.63 homologs and found that the GS5 strain behaves similarly to the UA159 strain, indicating that SprV's effect is strain independent.IMPORTANCE We previously identified a pleiotropic regulator, SprV, in Streptococcus mutans This regulator appears to be highly conserved among streptococci. Here, we showed that SprV regulates the expression of a secreted protein encoded by SMU.63 in S. mutans SMU.63 has been known to impact biofilm formation and genetic competence, two important characteristics that help in colonization of the organism. SMU.63 is also unique since it is known to form amyloid fiber. We found that SprV regulates the expression of SMU.63 at both the transcriptional and translational levels. We also found that the expression of SprV is regulated by magnesium ion concentration. Interestingly, both low and high magnesium ion concentrations affected biofilm formation and genetic competence. Since SMU.63 is also highly conserved among streptococci, we hypothesized that SprV will have a similar effect on its expression.


Quiescence Promotes Latent HIV Infection and Resistance to Reactivation from Latency with Histone Deacetylase Inhibitors.
Human immunodeficiency virus type 1 (HIV-1) establishes transcriptionally silent latent infections in resting memory T cells and hematopoietic stem and progenitor cells (HSPCs), which allows the virus to persist in infected individuals despite antiretroviral therapy. Developing in vitro models of HIV-1 latency that recapitulate the characteristics of latently infected cells in vivo is crucial to identifying and developing effective latency-reversing therapies. HSPCs exist in a quiescent state in vivo, and quiescence is correlated with latent infections in T cells. However, current models for culturing HSPCs and for infecting T cells in vitro require that the cells be maintained in an actively proliferating state. Here we describe a novel culture system in which primary human HSPCs cultured under hypothermic conditions are maintained in a quiescent state. We show that these quiescent HSPCs are susceptible to predominantly latent infection with HIV-1, while actively proliferating and differentiating HSPCs obtain predominantly active infections. Furthermore, we demonstrate that the most primitive quiescent HSPCs are more resistant to spontaneous reactivation from latency than more differentiated HSPCs and that quiescent HSPCs are resistant to reactivation by histone deacetylase inhibitors or P-TEFb activation but are susceptible to reactivation by protein kinase C (PKC) agonists. We also demonstrate that inhibition of HSP90, a known regulator of HIV transcription, recapitulates the quiescence and latency phenotypes of hypothermia, suggesting that hypothermia and HSP90 inhibition may regulate these processes by similar mechanisms. In summary, these studies describe a novel model for studying HIV-1 latency in human primary cells maintained in a quiescent state.IMPORTANCE Human immunodeficiency virus type 1 (HIV-1) establishes a persistent infection for which there remains no feasible cure. Current approaches are unable to clear the virus despite decades of therapy due to the existence of latent reservoirs of integrated HIV-1, which can reactivate and contribute to viral rebound following treatment interruption. Previous clinical attempts to reactivate the latent reservoirs in an individual so that they can be eliminated by the immune response or viral cytopathic effect have failed, indicating the need for a better understanding of the processes regulating HIV-1 latency. Here we characterize a novel in vitro model of HIV-1 latency in primary hematopoietic stem and progenitor cells isolated from human cord blood that may better recapitulate the behavior of latently infected cells in vivo This model can be used to study mechanisms regulating latency and potential therapeutic approaches to reactivate latent infections in quiescent cells.


Direct Sinus Lift and Immediate Implant Placement Using Piezosurgical Approach- A Case Report.
Numerous studies have shown that placement of implants in the maxillary region with resultant successful osseointegration can be achieved by the use of sinus lift procedures using piezosurgical technique. In this case report a middle aged patient had come to the outpatient department of A. J. Institute of Dental Sciences with a chief complaint of missing right posterior molar. Since the radiographic ball marker showed only 4 mm bone below the sinus in #16 region, direct sinus lift procedure was done for placement of a dental implant. Piezosurgery was performed, as it reduces the risk of damaging vital soft tissues such as nerves, dura matter and blood vessels. To stabilize the implant in the maxillary sinus region and also to stimulate bone regeneration, gamma irradiated cancellous allograft was used. Periapical radiographs were taken 10 months after implant placement which showed good bone growth over the implant collar. Bone formation in the maxillary antrum was seen clearly in the panoramic radiograph. Using piezosurgical unit, sinus lift procedure with sinus grafting proved to be less traumatic and more successful.


A Scoping Review to Map Empirical Evidence Regarding Key Domains and Questions in the Clinical Pathway of Delirium in Palliative Care.
Based on the clinical care pathway of delirium in palliative care (PC), a published analytic framework (AF) formulated research questions in key domains and recommended a scoping review to identify evidence gaps. To produce a literature map for key domains of the published AF: screening, prognosis and diagnosis, management, and the health-related outcomes. A standard scoping review framework was used by an interdisciplinary study team of nurse- and physician-delirium researchers, an information specialist, and review methodologists to conduct the review. Knowledge user engagement provided context in refining 19 AF questions. A peer-reviewed search strategy identified citations in Medline, PsycINFO, Embase, and CINAHL databases between 1980 and 2018. Two reviewers independently screened records for inclusion using explicit study eligibility criteria for the population, design, delirium diagnosis, and investigational intent. Of 104 studies reporting empirical data and meeting eligibility criteria, most were conducted in patients with cancer (73.1%) and in inpatient PC units (52%). The most frequent study design was a one or more group, nonrandomized trial or cohort (67.3%). Evidence gaps were identified: delirium risk prediction; comparative effectiveness and harms of prevention, variability in delirium management across PC settings, advanced directive and substitute decision-maker input, and transition of care location; and estimating delirium reversibility. Future rigorous primary studies are required to address these gaps and preliminary concerns regarding the quality of extant literature. Substantial evidence gaps exist, providing opportunities for future research regarding the assessment, prognosis, and management of delirium in PC settings.


Microbiological investigation of methane- and hydrocarbon-discharging mud volcanoes in the Carpathian Mountains, Romania.
Paclele Mici is a terrestrial mud volcano field located in the Carpathian Mountains (Romania), where thermal alteration of sedimentary organic compounds leads to methane, higher hydrocarbons and other petroleum compounds that are continuously released into the environment. The hydrocarbons represent potential substrates for microorganisms. We studied lipid biomarkers, stable isotope ratios, the effect of substrate (methane, other organic compounds) addition and 16S rRNA genes to gain insights into the hitherto unknown microbial community at this site. Quantitative real-time polymerase chain reaction analysis demonstrated that bacteria were much more abundant than archaea. Phylogenetic analyses of 16S rDNA clone sequences indicated the presence of bacterial and archaeal lineages generally associated with the methane cycle (methanogens, aerobic and anaerobic methanotrophs), the sulfur cycle (sulfate reducers), and groups linked to the anaerobic degradation of alkanes or aromatic hydrocarbons. The presence of sulfate reducers, methanogens and methanotrophs in this habitat was also confirmed by concurrent surveys of lipid biomarkers and their isotopic signatures. Incubation experiments with several common and complex substrates revealed the potential of the indigenous microbial community for sulfate reduction, methanogenesis and aerobic methanotrophy. Additionally, consistently to the detection of methane-oxidizing archaea (ANME) and 13C-depleted archaeal lipids, a weak but significant activity of anaerobic methane oxidation was measured by radiotracer techniques and in vitro. This survey is the first to report the presence and activity of ANME in a terrestrial environment.


Implementation of adaptive critic-based neurocontrollers for turbogenerators in a multimachine power system.
This paper presents the design and practical hardware implementation of optimal neurocontrollers that replace the conventional automatic voltage regulator (AVR) and the turbine governor of turbogenerators on multimachine power systems. The neurocontroller design uses a powerful technique of the adaptive critic design (ACD) family called dual heuristic programming (DHP). The DHP neurocontrollers' training and testing are implemented on the Innovative Integration M67 card consisting of the TMS320C6701 processor. The measured results show that the DHP neurocontrollers are robust and their performance does not degrade unlike the conventional controllers even when a power system stabilizer (PSS) is included, for changes in system operating conditions and configurations. This paper also shows that it is possible to design and implement optimal neurocontrollers for multiple turbogenerators in real time, without having to do continually online training of the neural networks, thus avoiding risks of instability.


New neotropical species of Opiinae (Hymenoptera, Braconidae) reared from fruit-infesting and leaf-mining Tephritidae (Diptera) with comments on the Diachasmimorpha mexicana species group and the genera Lorenzopius and Tubiformopius.
Four new species of opiine Braconidae are described from Mexico. These are Diachasmimorpha martinalujai Wharton reared from Rhagoletis infesting fruits of Crataegus spp., Diachasmimorpha norrbomi Wharton reared from Euphranta mexicana infesting fruits of Ribes pringlei, Eurytenes (Stigmatopoea) norrbomi Wharton reared from Trypeta concolor mining leaves of Barkleyanthus salicifolia and Eurytenes (Stigmatopoea) maya Wharton reared from Rhagoletis pomonella infesting apples and fruits of Crataegus spp. Morphological features of the first metasomal segment and occipital carina, useful for placement of these species, are discussed relative to the genera Diachasmimorpha, Eurytenes, Lorenzopius, Tubiformopius, and Opius s.l. Descriptions and diagnoses are referenced to the Hymenoptera Anatomy Ontology. The following represent new combinations: Diachasmimorpha hildagensis, Lorenzopius euryteniformis, and Tubiformopius tubibasis. Revised diagnoses are provided for Diachasmimorpha hildagensis, Diachasmimorpha mexicana, Diachasmimorpha sanguinea, Eurytenes (Stigmatopoea), Lorenzopius, Lorenzopius euryteniformis, Tubiformopius, Tubiformopius tubigaster, Tubiformopius tubibasis, Opius incoligma, and Opius rugicoxis. Two species groups are delineated within Lorenzopius and a key to species of Diachasmimorpha occurring in the New World is provided.


Susceptibility of monomicrobial or polymicrobial biofilms derived from infected diabetic foot ulcers to topical or systemic antibiotics in vitro.
Diabetic foot ulcers can become chronic and non-healing despite systemic antibiotic treatment. The penetration of systematically-administered antibiotics to the site of infection is uncertain, as is the effectiveness of such levels against polymicrobial biofilms. We have developed an in vitro model to study the effectiveness of different treatments for infected diabetic foot ulcers in a wound-like environment and compared the activity of systemic levels of antibiotics with that for topically applied antibiotics released from calcium sulfate beads. This is the first study that has harvested bacteria from diabetic foot infections and recreated similar polymicrobial biofilms to those present in vivo for individual subjects. After treatment with levels of gentamicin attained in serum after systemic administration (higher than corresponding tissues concentrations) we measured a 0-2 log reduction in bacterial viability of P. aeruginosa, S. aureus or a polymicrobial biofilm. Conversely, addition of gentamicin loaded calcium sulfate beads resulted in 5-9 log reductions in P. aeruginosa, S aureus and polymicrobial biofilms derived from three subjects. We conclude that systemically administered antibiotics are likely to be inadequate for successfully treating these infections, especially given the vastly increased concentrations required to inhibit cells in a biofilm, and that topical antibiotics provide a more effective alternative.


Perspectives of US Youth During Initial Month of the COVID-19 Pandemic.
Asymptomatic youth in the United States acting as "silent spreaders" during the coronavirus disease 2019 (COVID-19) pandemic are an ongoing public health concern, particularly given their depiction as unengaged with recommendations. Our goal was to understand the knowledge, beliefs, and experiences of US youth at the onset of the COVID-19 pandemic. We posed 2 open-ended surveys to the national MyVoice text message cohort of youth, aged 14-24 years. On March 6, 2020, 4 questions were asked regarding knowledge and experiences during the COVID-19 pandemic with 3 questions repeated on March 20, 2020. Qualitative responses were coded using thematic analysis and summarized with descriptive statistics. Of 1,174 youth, 1,087 responded to at least 1 question (response rate of 88%). The average age of respondents was 19 (SD 2.8) years with 52% female and 56% non-Hispanic White respondents. On March 6, 2020, most (70%) respondents reported knowing about COVID-19 and primarily cited the news (46%) as their source of information. Nearly all (95%) respondents reported impact by March 20, 2020, and respondents expressing worry increased from 25% to 51%. In both surveys, worried youth primarily cited concern for others (26% and 34%). Regarding preparation, respondents primarily reported doing nothing (36%) on March 6, 2020, and practicing social distancing (50%) on March 20, 2020. Many youths in our sample are engaged with the COVID-19 pandemic and most are feeling knowledgeable, are concerned about its impacts on others, and are practicing social distancing. Sustained public health efforts should focus on maintaining youth engagement with accurate public information and youth-centered messaging promoting prevention measures to protect the health and well-being of youth and their friends and family.


Transcytotic vesicular carriers for polymeric IgA receptors accumulate in rat hepatocytes after bile duct ligation.
In rat hepatocytes, transcytotic vesicular carriers transport the mature 120 x 10(3) Mr form of the polymeric IgA receptor (pIgA-R), with or without its ligand, pIgA, from the sinusoidal to the biliary plasmalemma, where the ectodomain of the receptor is cleaved to produce an 80 x 10(3) Mr fragment that is secreted into the bile. Here we show that cholestasis induced by bile duct ligation results in the accumulation of transcytotic carriers, identified by the 120 x 10(3) Mr pIgA-R and pIgA, in the pericanalicular cytoplasm of hepatocytes. To determine the extent of pIgA-R accumulation, hepatic total microsomes (TM) were prepared from control and cholestatic rats. Solubilized TM proteins were separated by SDS-PAGE and receptor forms were detected by immunoblotting and autoradiography. Quantitative densitometry of these autoradiograms showed that after duct ligation the 120 x 10(3) Mr receptor accumulated to a level approximately threefold higher than the control. Concomitantly, immunologically related, novel 124, 90 and 80 x 10(3) Mr proteins (cholestatic antigens) became detectable. Immunoblot analyses of biliary and serum proteins showed that cholestasis resulted in: (1) a marked decrease in the concentrations of the 80 x 10(3) Mr receptor and pIgA in the bile, whereas albumin concentrations remained at control levels; and (2) a marked increase in the concentration of the 80 x 10(3) Mr receptor in the serum. Positive sites for pIgA-R were localized to the pericanalicular cytoplasm of hepatocytes by indirect immunofluorescence on semithin frozen sections in cholestatic hepatocytes. The sites were more numerous and the positive signal stronger than in controls. One day post-ligation, pIgA-positive sites were located to the same pericanalicular cytoplasm of hepatocytes; by three days, however, most pIgA appeared in sinusoidal endothelia and Kupffer cells. To validate the vesicular character of the receptor-positive sites, sham-operated and cholestatic livers were processed for either transmission electron microscopy (TEM) or immunogold localization of receptors on thin frozen sections. TEM verified the accumulation of pericanalicular vesicles in cholestatic hepatocytes. Immunogold tests localized pIgA-R to pleiomorphic, pericanalicular vesicles, which were increased in number, size and concentration of antigenic sites in cholestatic hepatocytes. These findings indicate that bile duct ligation provides a method for manipulating the in vivo transcytotic pathway and for accumulating previously unstudied transcytotic carriers in hepatocytes.


[Ontogenic properties of peptidohydrolase activity in tissue extracts from Drosophila melanogaster].
The paper deals with the activity of peptidohydrolases splitting hemoglobin, casein and benzoylarginine-p-nitroanilide (BAPNA) in the ontogenesis of Drosophila mutant white apricot (wa). The enzymic activity is high in the intestine tissue at the larval stage, but it does not manifest itself at the pupal stage that may be due to inactivation of the studied enzymes by inhibitors found in extracts of the pupal tissues. The imaginal period of development is characterized by a low level of the peptidohydrolase activity in females and its absence in males in whose tissues there is a high content of inhibitors. The peptidohydrolase activity in extracts of larval tissues is established to be dependent completely on the enzyme being in the intestinal wall. Incomplete purification of this peptidohydrolase and its inhibitor is performed by means of ion-exchange chromatography on DEAE-cellulose. Some of their physicochemical properties are investigated.


Organogenesis from transformed tomato explants.
Tomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transformation and the technique has been useful for the isolation and identification of many genes involved in plant disease resistance, morphology and development. In this method, cotyledon explants from in vitro-grown seedlings are precultured overnight on a tobacco suspension feeder layer. The explants are then inoculated with Agrobacterium and returned to the feeder layer for a 2-d period of cocultivation. After cocultivation, the explants are transferred to an MS-based selective regeneration medium containing zeatin. Regenerated shoots are then rooted on a separate selective medium. This protocol has been used with several tomato cultivars and routinely yields transformation efficiencies of 10-15%.


Klinefelter's syndrome associated with mediastinal germ cell neoplasms.
Several case reports have suggested an association of primary mediastinal germ cell tumor (PMGCT) and Klinefelter's syndrome (KS). In an effort to confirm this association, 22 patients with mediastinal germ cell tumors had chromosome studies performed in a prospective fashion. Five patients (22%) had karyotypic or pathologic evidence of KS. All of the patients with KS had germ cell tumors of the nonseminomatous subtype and were relatively young (median age, 15 years). The literature confirms the findings of a young median age (18 years), nonseminomatous subtype, and mediastinal location of the germ cell neoplasm. We conclude that patients with KS are predisposed to the development of mediastinal nonseminomatous germ cell cancers.


Domain evolution and functional diversification of sulfite reductases.
Sulfite reductases are key enzymes of assimilatory and dissimilatory sulfur metabolism, which occur in diverse bacterial and archaeal lineages. They share a highly conserved domain "C-X5-C-n-C-X3-C" for binding siroheme and iron-sulfur clusters that facilitate electron transfer to the substrate. For each sulfite reductase cluster, the siroheme-binding domain is positioned slightly differently at the N-terminus of dsrA and dsrB, while in the assimilatory proteins the siroheme domain is located at the C-terminus. Our sequence and phylogenetic analysis of the siroheme-binding domain shows that sulfite reductase sequences diverged from a common ancestor into four separate clusters (aSir, alSir, dsr, and asrC) that are biochemically distinct; each serves a different assimilatory or dissimilatory role in sulfur metabolism. The phylogenetic distribution and functional grouping in sulfite reductase clusters (dsrA and dsrB vs. aSiR, asrC, and alSir) suggest that their functional diversification during evolution may have preceded the bacterial/archaeal divergence.


Enhancing Tax Compliance through Coercive and Legitimate Power of Tax Authorities by Concurrently Diminishing or Facilitating Trust in Tax Authorities.
Both coercion, such as strict auditing and the use of fines, and legitimate procedures, such as assistance by tax authorities, are often discussed as means of enhancing tax compliance. However, the psychological mechanisms that determine the effectiveness of each strategy are not clear. Although highly relevant, there is rare empirical literature examining the effects of both strategies applied in combination. It is assumed that coercion decreases implicit trust in tax authorities, leading to the perception of a hostile antagonistic tax climate and enforced tax compliance. Conversely, it is suggested that legitimate power increases reason-based trust in the tax authorities, leading to the perception of a service climate and eventually to voluntary cooperation. The combination of both strategies is assumed to cause greater levels of intended compliance than each strategy alone. We conducted two experimental studies with convenience samples of 261 taxpayers overall. The studies describe tax authorities as having low or high coercive power (e.g., imposing lenient or severe sanctions) and/or low or high legitimate power (e.g., having nontransparent or transparent procedures). Data analyses provide supportive evidence for the assumptions regarding the impact on intended tax compliance. Coercive power did not reduce implicit trust in tax authorities; however, it had an effect on reason-based trust, interaction climate, and intended tax compliance if applied solely. When wielded in combination with legitimate power, it had no effect.


[Computed tomography of the temporal bone in diagnosis of chronic exudative otitis media].
Computed tomography (CT) of the temporal bone was made in 37 patients aged 2 to 55 years with chronic exudative otitis media (CEOM). In 21 of them the pathology was bilateral. The analysis of 58 CT images has identified CT signs of chronic exudative otitis media. They include partial (17 temporary bones) or complete (38 temporal bones) block of the bone opening of the auditory tube, pneumatic defects of the tympanic cavity (58 temporal bones), pneumatic defects of the mastoid process and antrum (47 temporal bones), pathologic retraction of the tympanic membrane. The examination of the temporal bone detected both CT-signs of CEOM and other causes of hearing disorders in 14 patients (26 temporal bones) with CEOM symptoms and inadequately high hypoacusis. Among these causes were malformation of the auditory ossicula (n=5), malformation of the labynthine window (n=2), malformation of the middle and internal ear (n=4), a wide aqueduct of the vestibule, labyrinthine anomaly of Mondini's type (n=1), cochlear hypoplasia (n=4), stenosis of the internal acoustic meatuses (n=2). Sclerotic fibrous dysplasia was suggested in 2 temporal bones (by CT data). CT was repeated after surgical treatment of 10 patients (14 temporal bones) and visual assessment of tympanostomy results was made.


Adjuvant chemotherapy for breast cancer: side effects and quality of life.
In a trial of postoperative adjuvant chemotherapy women with primary breast cancer and spread to one or more axillary nodes were randomised to receive a six-month course of either the single agent chlorambucil or the five-drug combination of chlorambucil, methotrexate, fluorouracil, vincristine, and adriamycin. On completing the treatment 47 patients were asked to fill in questionnaires at home on the side effects of treatment and its influence on the quality of their life. Side effects including nausea, vomiting, malaise, and alopecia had been severe enough to interfere with their lifestyle in 9 (42%) of the patients who had received the single agent and 19 (79%) of those who had received multiple-drug treatment. Various other side effects were reported by a few patients. Seven (29%) of the patients who had received the multiple-drug schedule voluntarily added that the treatment had been "unbearable" or "could never be gone though again." The proportion of patients who had experienced severe side effects while receiving the treatment was considerable; hence such adjuvant chemotherapy is justifiable only if it will substantially improve a patient's prognosis.


Telomere dysfunction cooperates with epigenetic alterations to impair murine embryonic stem cell fate commitment.
The precise relationship between epigenetic alterations and telomere dysfunction is still an extant question. Previously, we showed that eroded telomeres lead to differentiation instability in murine embryonic stem cells (mESCs) via DNA hypomethylation at pluripotency-factor promoters. Here, we uncovered that telomerase reverse transcriptase null (Tert-/-) mESCs exhibit genome-wide alterations in chromatin accessibility and gene expression during differentiation. These changes were accompanied by an increase of H3K27me3 globally, an altered chromatin landscape at the Pou5f1/Oct4 promoter, and a refractory response to differentiation cues. Inhibition of the Polycomb Repressive Complex 2 (PRC2), an H3K27 tri-methyltransferase, exacerbated the impairment in differentiation and pluripotency gene repression in Tert-/- mESCs but not wild-type mESCs, whereas inhibition of H3K27me3 demethylation led to a partial rescue of the Tert-/- phenotype. These data reveal a new interdependent relationship between H3K27me3 and telomere integrity in stem cell lineage commitment that may have implications in aging and cancer.


Effect of music on chronic osteoarthritis pain in older people.
Osteoarthritis is the most common degenerative disease in humans. It usually begins in middle age and is progressive. Chronic pain in older people presents a significant obstacle in maintaining function and independence. Previous studies have shown that music can improve motivation, elevate mood, and increase feelings of control in older people. The purpose of this randomized clinical trial was to examine the influence of music as a nursing intervention on osteoarthritis pain in elders. Data were collected using the short form of the McGill Pain Questionnaire with 66 elders suffering from chronic osteoarthritis pain. Differences in perceptions of pain were measured over 14 days in an experimental group who listened to music for 20 minutes daily and a control group who sat quietly for 20 minutes daily. All participants completed the Short Form McGill Pain Questionnaire (SF-MPQ) on day 1, 7, and 14 of the study. Results of t-tests indicated that those who listened to music had less pain on both the Pain Rating Index on day 1 (P = 0.001), day 7 (P = 0.001) and day 14 (P = 0.001) and on the Visual Analogue Scale on day 1 (P = 0.001), day 7 (P = 0.001) and day 14 (P = 0.001), when compared with those who sat quietly and did not listen to music. A repeated measure analysis of variance controlling for pretest measures demonstrated a significant decrease in pain among experimental group participants when compared with the control group on the pain descriptor section of the SF-MPQ (P = 0.001) and the visual analogue portion of the SF-MPQ (P = 0.001). Listening to music was an effective nursing intervention for the reduction of chronic osteoarthritis pain in the community-dwelling elders in this study.


Bullet points to evaluate the performance of the middle-down proteomics workflow for histone modification analysis.
Middle-down proteomics has emerged as the method of choice to study combinatorial histone post translational modifications (PTMs). In the common bottom-up workflow, histones are digested into relatively short peptides (4-20 aa), separated using reversed-phase chromatography and analyzed using typical proteomics methods in mass spectrometry. In middle-down, histones are cleaved into longer polypeptides (50-60 aa) mostly corresponding to their N-terminal tails, resolved using weak cation exchange-hydrophilic interaction liquid chromatography (WCX-HILIC) and analyzed with less conventional mass spectrometry, i.e. using Electron Transfer Dissociation (ETD) for analyte fragmentation. Middle-down is not nearly as utilized as bottom-up for PTM analysis, partially due to its limited reproducibility and robustness. This has also limited the establishment of rigorous benchmarks to discriminate good vs poor quality experiments. Here, we describe critical aspects of the middle-down workflow to assist the user in evaluating the presence of biased and misleading results. Specifically, we tested the use of porous graphitic carbon (PGC) during the desalting step, demonstrating that desalting using only C18 material leads to sample loss. We also tested different salts in the WCX-HILIC buffers for their effect on retention, selectivity, and reproducibility of analysis of variants of histone tail fragments, in particular replacing ammonium ion with ethylenediammonium ion in buffer A. These substitutions had marked effects on selectivity and retention. Our results provide a streamlined way to evaluate middle-down performance to identify and quantify combinatorial histone PTMs.


The content of sexual fantasies for sexual offenders.
Although the phenomenon of sexual fantasy has been extensively researched, little contemporary inquiry has investigated the content of sexual fantasy within the context of sexual offending. In this study, a qualitative analysis was used to develop a descriptive model of the phenomena of sexual fantasy during the offence process. Twenty-four adult males convicted of sexual offences provided detailed retrospective descriptions of their thoughts, emotions, and behaviors before, during, and after their offences. Using Grounded Theory, a model was developed to elucidate the content and themes of sexual fantasy for sexual offenders, as well as the way fantasy content was used in the process of sexual offending. The Sexual Fantasy Content Model (SFCM) comprises of three higher-order (level 1) and five second-order (level 2) categories that describe the content of sexual fantasy across the offence process. The level 1 categories are general sexual fantasy, nonspecific offence fantasy, and offence-specific fantasy, whereas the level 2 categories included demographic, behavioral, relational, situational, and self-perceptual considerations. The strengths of the SFCM are discussed and its clinical implications are reviewed.


New evidence on the importance of the renin-angiotensin system in the treatment of higher-risk patients with hypertension.
We reviewed the drug treatment of hypertension in the light of recent trials. beta-Blockers and diuretics clearly reduce mortality, strokes, and coronary heart disease (CHD) in hypertension. Recent trials assessed whether newer agents that block the renin-angiotensin-aldosterone system, or calcium blockers, offer any additional advantage, or have benefits in high-risk individuals with conventionally 'normal' blood pressure. The recent ALLHAT study claimed no differences in CHD or mortality when chlorthalidone, amlodipine, and lisinopril were compared. However, the decrease in blood pressure was not the same with the three agents, and a substantial proportion of patients enrolled did not have clinical disease. In contrast, the LIFE study (comparing losartan and a beta-blocker) and the ANBP-2 study [comparing angiotensin-converting enzyme (ACE) inhibition and a diuretic] reduced blood pressure similarly, yet demonstrated benefits in favour of angiotensin II type 1 receptor blockers (ARBs) and ACE inhibitors. Other trials indicated similar advantages of ACE inhibitors or ARBs in patients with diabetic nephropathy. Among high-risk patients with initial blood pressure in the 'normal' range, ACE inhibitors significantly reduce clinical events (mortality, strokes, and myocardial infarction), despite modest decreases in blood pressure, suggesting that additional mechanisms are responsible. Recent results of the Prospective Studies Collaboration show lower risk, even in the normal blood pressure range; high-risk patients will benefit further from ACE inhibitors and ARBs (and beta-blockers after myocardial infarction). Data for other blood pressure decreasing agents are unavailable in such populations. We conclude that blood pressure decreasing per se is of clinical benefit, but drugs that block the renin-angiotensin system offer additional advantages. Drug choice is best determined by the patient's clinical condition.


Deciphering Patterns of Adaptation and Acclimation in the Transcriptome of Phaeocystis antarctica to Changing Iron Conditions1.
The haptophyte Phaeocystis antarctica is endemic to the Southern Ocean, where iron supply is sporadic and its availability limits primary production. In iron fertilization experiments, P. antarctica showed a prompt and steady increase in cell abundance compared to heavily silicified diatoms along with enhanced colony formation. Here we utilized a transcriptomic approach to investigate molecular responses to alleviation of iron limitation in P. antarctica. We analyzed the transcriptomic response before and after (14 h, 24 h and 72 h) iron addition to a low-iron acclimated culture. After iron addition, we observed indicators of a quick reorganization of cellular energetics, from carbohydrate catabolism and mitochondrial energy production to anabolism. In addition to typical substitution responses from an iron-economic toward an iron-sufficient state for flavodoxin (ferredoxin) and plastocyanin (cytochrome c6 ), we found other genes utilizing the same strategy involved in nitrogen assimilation and fatty acid desaturation. Our results shed light on a number of adaptive mechanisms that P. antarctica uses under low iron, including the utilization of a Cu-dependent ferric reductase system and indication of mixotrophic growth. The gene expression patterns underpin P. antarctica as a quick responder to iron addition.


Glucagon-mediated impairments in hepatic and peripheral tissue nutrient disposal are not aggravated by increased lipid availability.
Glucose, fat, and glucagon availability are increased in diabetes. The normal response of the liver to chronic increases in glucose availability is to adapt to become a marked consumer of glucose. Yet this fails to occur in diabetes. The aim was to determine whether increased glucagon and lipid interact to impair the adaptation to increased glucose availability. Chronically catheterized well controlled depancreatized conscious dogs (n = 21) received 3 days of continuous parenteral nutrition (TPN) that was either high in glucose [C; 75% nonprotein calories (NPC)] or in lipid (HL; 75% NPC) in the presence or absence of a low dose (one-third basal) chronic intraportal infusion of glucagon (GN; 0.25 ng.kg(-1).min(-1)). During the 3 days of TPN, all groups received the same insulin algorithm; the total amount of glucose infused (GIR) was varied to maintain isoglycemia ( approximately 120 mg/dl). On day 3 of TPN, hepatic metabolism was assessed. Glucose and insulin levels were similar in all groups. GIR was decreased in HL and C + GN ( approximately 30%) and was further decreased in HL + GN (55%). Net hepatic glucose uptake was decreased approximately 15% in C + GN, and HL and was decreased approximately 50% in HL + GN. Lipid alone or combined with glucagon decreased glucose uptake by peripheral tissues. Despite impairing whole body glucose utilization, HL did not limit whole body energy disposal. In contrast, glucagon suppressed whole body energy disposal irrespective of the diet composition. In summary, failure to appropriately suppress glucagon secretion adds to the dietary fat-induced impairment in both hepatic and peripheral glucose disposal. In addition, unlike increasing the percentage of calories as fat, inappropriate glucagon secretion in the absence of compensatory hyperinsulinemia limits whole body nutrient disposition.


Study of the relationship between tuberous sclerosis complex and autistic disorder.
There has been increasing awareness that there are behavioral phenotypes in tuberous sclerosis complex with neuropsychiatric symptom complex such as autistic disorder and attention-deficit hyperactivity disorder (ADHD). However, the neurobiologic basis of autistic disorder in tuberous sclerosis complex is still unknown. We studied two cohorts of children followed up since 1986 until 2003, one cohort with tuberous sclerosis complex and another cohort with autistic disorder, to determine the incidence of autistic disorder in tuberous sclerosis complex and the incidence of tuberous sclerosis complex in autistic disorder respectively. We established a Tuberous Sclerosis Complex Registry in 1985 at the University of Hong Kong. In 2004, 44 index cases (the male to female ratio was 0.75:1) were registered. Three had a positive family history of tuberous sclerosis complex. Thus, the total number of tuberous sclerosis complex cases was 47. We adopted the diagnostic criteria of tuberous sclerosis complex for case ascertainment. The period prevalence rate of tuberous sclerosis complex for children and adolescents aged < 20 years is 3.5 per 10,000 (on Hong Kong island, excluding the eastern region with 125,100 aged < 20 years in 2003). Of 44 cases with tuberous sclerosis complex, 7 had autistic disorder. Thus, the incidence of autistic disorder in tuberous sclerosis complex is 16%. During the 17-year period (1986-2003), we collected a database of 753 children (668 boys and 84 girls; male to female ratio 8:1) with autistic disorder and pervasive developmental disorders. For all children with autistic disorder or pervasive developmental disorders, we routinely examined for any features of tuberous sclerosis complex by looking for neurocutaneous markers such as depigmented spots, which appear in 50% of children with tuberous sclerosis complex by the age of 2 years. For those with infantile spasm or epilepsy, the clinical features of tuberous sclerosis complex were monitored regularly during follow-up. Of these, seven had tuberous sclerosis complex. Thus, the incidence of tuberous sclerosis complex in autistic disorder is 0.9%. All of these children are mentally retarded, with moderate to severe grades in an intellectual assessment conducted by a clinical psychologist. Future studies should be directed toward looking at the various behavioral phenotypes in tuberous sclerosis complex and defining these with standardized criteria to look for any real association with the underlying genetic mutation of TSC1 or TSC2 gene or even the site of tubers in the brain.


Evaluation of the BBL Minitek system for the identification of Enterobacteriaceae.
Thirty-one different substrate disks were tested in parallel with comparable, prepared media (BBL) against a minimum of 300 cultures of Enterobacteriaceae. An overall correlation of 98% was observed with all the disks tested. In addition, the system was used to identify 461 fresh isolates of Enterobacteriaceae in parallel with conventional media using the schema used at the Veterans Administration Hospital, Baltimore. An overall correlation of 97% was observed. Minitek is a time and space saving system. It is accurate and easily adapted to the clinical laboratory. A wide variety of substrates are available, allowing most laboratories to use their own schema. The long shelf life of most disks is a definite advantage.


A single internalization signal from the di-leucine family is critical for constitutive endocytosis of the type II TGF-beta receptor.
Endocytosis has an important contribution to the regulation of the surface expression levels of many receptors. In spite of the central role of the transforming growth factor beta (TGF-beta) receptors in numerous cellular and physiological processes, their endocytosis is largely unexplored. Current information on TGF-beta receptor endocytosis relies exclusively on studies with chimeric constructs containing the extracellular domain of the GM-CSF receptors, following the internalization of the GM-CSF ligand; the conformation and interactions of the chimeric receptors (and therefore their endocytosis) may differ considerably from those of the native TGF-beta receptors. Furthermore, there are no data on the potential endocytosis motif(s) of the TGF-beta receptors or other receptor Ser/Thr kinases. Here, we report the use of type II TGF-beta receptors, myc-tagged at their extracellular terminus, to investigate their endocytosis. Employing fluorescent antibody fragments to label exclusively the cell surface myc-tagged receptors exposed to the external milieu, made it possible to follow the internalization of the receptors, without the complications that render labeling with TGF-beta (which binds to many cellular proteins) unsuitable for such studies. The results demonstrate that the full-length type II TGF-beta receptor undergoes constitutive endocytosis via clathrin-coated pits. Using a series of truncation and deletion mutants of this receptor, we identified a short peptide sequence (I(218)I(219)L(220)), which conforms to the consensus of internalization motifs from the di-leucine family, as the major endocytosis signal of the receptor. The functional importance of this sequence in the full-length receptor was validated by the near complete loss of internalization upon mutation of these three amino acids to alanine.


Enhanced urinary immunoreactive thromboxane in neonatal necrotizing enterocolitis. A diagnostic indicator of thrombotic activity.
Urinary thromboxane B2 levels increased threefold to 20-fold in infants with neonatal necrotizing enterocolitis compared with healthy infants and infants with benign causes of heme-positive stools. Increased urinary thromboxane B2 levels were detected coincidently with the initial signs of necrotizing enterocolitis, and values paralleled the course of the illness. Infants with serious illnesses other than necrotizing enterocolitis had increased urinary thromboxane B2 levels but with lower values than those of infants with necrotizing enterocolitis. Indomethacin therapy appeared to reduce urinary thromboxane B2 levels and reduce their usefulness as a marker of illness. Another product of platelet activation, beta-thromboglobulin, was increased in the urine of infants with necrotizing enterocolitis. Decreased platelet counts in infants with necrotizing enterocolitis correlated inversely with urinary thromboxane. Results of beta-thromboglobulin and platelet studies are consistent with the concept that platelet consumption due to ischemic thrombosis was the source of enhanced thromboxane excretion.


Studies with anthracyclines in pediatric acute nonlymphocytic leukemia.
The anthracyclines, daunomycin and Adriamycin, have become two of the most important chemotherapeutic agents in the treatment of pediatric acute nonlymphocytic leukemia. Data are presented on 306 children treated after initial diagnosis with an anthracycline or one of the chemotherapeutic agents. The remission rate was 65%. When not used during maintenance therapy, the anthracyclines have been shown to be effective in inducing a second remission even though the drug was used during initial therapy. An ongoing randomized study utilizing Adriamycin in induction of remission and during maintenance therapy is also discussed.


Chronological and subjective age in relation to work demands: survey of Croatian workers.
The aim of the study was to investigate chronological and subjective age and gender differences in terms of the physical, mental, and social demands of work. Subjects, 88 women and 25 men, were workers in a medical setting who completed the Survey of Health Care Professionals. Subjects' reports of effort required by their work, tiredness resulting from their work, and personal skills needed to complete their work were used to assess the physical, mental, and social demands of work. Subjective age measures included cognitive age, desired age, and comparative age. Results showed that, compared to men, women reported being more tense as a result of the work. Age differences were found in the physical and mental skills required by the work. Older workers had lower estimates of their own skills than younger workers. Three measures of subjective age showed different patterns of relationships with aspects of work demands.


Piloting of an oral health education program and knowledge test for midwives.
Abstract Research shows limited emphasis being placed on oral health by midwives in Australia and the need for further education in this area. The study aim was to pilot a midwifery oral health education program and knowledge test and identify any flaws in its content and design. Twenty two midwives from an antenatal ward in South-western Sydney completed the program and 12 feedback forms/knowledge tests were returned. Data was analysed using descriptive statistics and content analysis. Feedback data showed all midwives appreciated that the program was available online and self-paced. Most found the program extremely informative and following completion were more confident in promoting maternal oral health. The mean correct responses in the knowledge test was 79 % (SD=12.3) which suggests most items were suitable for assessing knowledge improvement. However, in 3 items midwives had low correct responses. Various aspects that could be improved or clarified were identified and suggestions discussed.


Meckel's diverticulum: the false-negative examination.
A case of Meckel's diverticulum is presented in a 17-month-old girl following technetium pertechnetate radioisotope scan. The patient had presented 10 months earlier with the first episode of passing bloody stools. A Meckel's diverticulum was resected during surgery. Reasons for the original false-negative examination are presented.


Spatial distribution of microplastics around an inhabited coral island in the Maldives, Indian Ocean.
Small plastic particles are considered environmental pollutants and are highly concentrated in marine sediments. However, knowledge about plastic abundance within coral reef habitat and beach sediments surrounding remote inhabited coral islands is scarce. In this study, microplastic accumulation was investigated on a small inhabited coral island located in the Maldives. Sediments from 22 sampling sites across fore reef, reef flat, and beach environments were analysed for plastic particles <5 mm. Density separation and microscope enumeration revealed a total of 1244 individual microplastic pieces, in filamentous (49%) and fragmented (51%) forms, found across all sampling sites. High concentrations were recorded at all sites, however, there was no significant relationship between microplastic concentration or size across regions (inner atoll and outer atoll) or environments (fore reef, reef flat, and beach). Furthermore, concentrations of microplastic fragment and filament forms, total concentration, and the microplastic community, showed little correlation with sediment particle size. Our findings show microplastics are ubiquitous in marine sediments around a remote coral island, at sizes ingestible by marine organisms, raising concerns about potential effects of microplastic ingestion by coral reef species.


Investigating the Structure of Neurotoxic Protein Aggregates Inside Cells.
Neurodegenerative diseases affect the lives of millions of people across the world, being particularly prevalent in the aging population. Despite huge research efforts, conclusive insights into the disease mechanisms are still lacking. Therefore, therapeutic strategies are limited to symptomatic treatments. A common histopathological hallmark of many neurodegenerative diseases is the presence of large pathognomonic protein aggregates, but their role in the disease pathology is unclear and subject to controversy. Here, we discuss imaging methods allowing investigation of these structures within their cellular environment: conventional electron microscopy (EM), super-resolution light microscopy (SR-LM), and cryo-electron tomography (cryo-ET). Multidisciplinary approaches are key for understanding neurodegenerative diseases and may contribute to the development of effective treatments. For simplicity, we focus on huntingtin aggregates, characteristic of Huntington's disease.