The primary transcriptome of the marine diazotroph Trichodesmium erythraeum IMS101.
Blooms of the dinitrogen-fixing marine cyanobacterium Trichodesmium considerably contribute to new nitrogen inputs into tropical oceans. Intriguingly, only 60% of the Trichodesmium erythraeum IMS101 genome sequence codes for protein, compared with ~85% in other sequenced cyanobacterial genomes. The extensive non-coding genome fraction suggests space for an unusually high number of unidentified, potentially regulatory non-protein-coding RNAs (ncRNAs). To identify the transcribed fraction of the genome, here we present a genome-wide map of transcriptional start sites (TSS) at single nucleotide resolution, revealing the activity of 6,080 promoters. We demonstrate that T. erythraeum has the highest number of actively splicing group II introns and the highest percentage of TSS yielding ncRNAs of any bacterium examined to date. We identified a highly transcribed retroelement that serves as template repeat for the targeted mutation of at least 12 different genes by mutagenic homing. Our findings explain the non-coding portion of the T. erythraeum genome by the transcription of an unusually high number of non-coding transcripts in addition to the known high incidence of transposable elements. We conclude that riboregulation and RNA maturation-dependent processes constitute a major part of the Trichodesmium regulatory apparatus.


Mechanistic analysis of an asymmetric palladium-catalyzed conjugate addition of arylboronic acids to β-substituted cyclic enones.
An asymmetric palladium-catalyzed conjugate addition reaction of arylboronic acids to enone substrates was investigated mechanistically. Desorption electrospray ionization coupled to mass spectrometry was used to identify intermediates of the catalytic cycle and delineate differences in substrate reactivity. Our findings provide evidence for the catalytic cycle proceeding through formation of an arylpalladium(II) cation, subsequent formation of an arylpalladium-enone complex, and, ultimately, formation of the new C-C bond. Reaction monitoring in both positive and negative ion modes revealed that 4-iodophenylboronic acid formed a relatively stable trimeric species under the reaction conditions.


The Clinical Significance of HPV Type in Women with Recurrent Cytological Atypia.
The human papillomavirus (HPV) test has proven to be efficient in triaging women with abnormal Pap findings in women with low cytological atypia, but there is no data about the accuracy for large loop excision of transformation zone in cases of recurrent atypia. To assess the clinical correlation between results of HPV typing and conization histology in women who had recurrent abnormal Pap test results with no colposcopy findings. Our retrospective cohort study included 138 women enrolled in the Maccabi Healthcare Services who had consecutive atypical Pap test results for 2 years in which no abnormal colposcopic findings were detected. These women had an HPV typing and then conization. Among the total study population (n=138), 71.7% had negative histology, 19.6% had ≤ cervical intraepithelial neoplasia grade 1 (≤ CIN1), and 8.7% had CIN2+. With regard to HPV typing, 34.8% were negative and 65.2% were positive. Of those testing positive, 34.4% were positive for HPV 16 or 18. Sensitivity, specificity, positive predictive value, and negative predictive values of HPV typing for women were 89.7%, 44.4%, 38.9%, and 91.7%, respectively, and for HPV 16 or 18: 71.4%, 67.7%, 32.3%, and 100.0%, respectively. After stratification by cytological grades, for women with high-grade cervical cytology, the sensitivity and negative predictive values of the HPV typing were higher than among low-grade cervical cytology, while specificity and positive predictive values were lower. HPV typing is a useful tool for the management of patients with persistently abnormal Pap test results.


Amber-Compatible Parametrization Procedure for Peptide-like Compounds: Application to 1,4- and 1,5-Substituted Triazole-Based Peptidomimetics.
Peptidomimetics are molecules of particular interest in the context of drug design and development. They are proteolytically and metabolically more stable than their natural peptide counterparts but still offer high specificity toward their biological targets. In recent years, 1,4- and 1,5-disubstituted 1,2,3-triazole-based peptidomimetics have emerged as promising lead compounds for the design of various inhibitory and tumor-targeting molecules as well as for the synthesis of peptide analogues. The growing popularity of triazole-based peptidomimetics and a constantly broadening range of their application generated a demand for elaborate theoretical investigations by classical molecular dynamics simulations and molecular docking. Despite this rising interest, accurate and coherent force field parameters for triazole-based peptidomimetics are still lacking. Here, we report the first complete set of parameters dedicated to this group of compounds, named TZLff. This parametrization is compatible with the latest version of the AMBER force field (ff14SB) and can be readily applied for the modeling of pure triazole-based peptidomimetics as well as natural peptide sequences containing one or more triazole-based modifications in their backbone. The parameters were optimized to reproduce HF/6-31G* electrostatic potentials as well as MP2/cc-pVTZ equilibrium Hessian matrices and conformational potential energy surfaces through the use of a genetic algorithm-based search and least-squares fitting. Following the standards of AMBER, we introduce residue building units, thus allowing the user to define any given sequence of triazole-based peptidomimetics. Validation of the parameter set against ab initio- and NMR-based reference systems shows that we obtain fairly accurate results, which properly capture the conformational features of triazole-based peptidomimetics. The successful and efficient parametrization strategy developed in this work is general enough to be applied in a straightforward manner for parametrization of other peptidomimetics and, potentially, any polymeric assemblies.


Rapid recovery of Escherichia coli from estuarine water.
The efficiencies of two 24-hr elevated-temperature tests to recover Escherichia coli from estaurine water were compared simultaneously with the 72-hr standard methods procedure of the American Public Health Association (APHA). From 1,710 tubes, E. coli was recovered 222 times in lauryl tryptose medium incubated at 44 +/- 0.2 C for 24 hr, 261 times in an experimental medium incubated at 44.5 +/- 0.2 C for 24 hr, and 257 times by the 72-hr APHA method. The number of false positives enumerated was similar in all three tests. The data indicated that E. coli in raw seawater could be determined in 24 hr without a significant loss of accuracy.


Expression of Tas1 taste receptors in mammalian spermatozoa: functional role of Tas1r1 in regulating basal Ca²⁺ and cAMP concentrations in spermatozoa.
During their transit through the female genital tract, sperm have to recognize and discriminate numerous chemical compounds. However, our current knowledge of the molecular identity of appropriate chemosensory receptor proteins in sperm is still rudimentary. Considering that members of the Tas1r family of taste receptors are able to discriminate between a broad diversity of hydrophilic chemosensory substances, the expression of taste receptors in mammalian spermatozoa was examined. The present manuscript documents that Tas1r1 and Tas1r3, which form the functional receptor for monosodium glutamate (umami) in taste buds on the tongue, are expressed in murine and human spermatozoa, where their localization is restricted to distinct segments of the flagellum and the acrosomal cap of the sperm head. Employing a Tas1r1-deficient mCherry reporter mouse strain, we found that Tas1r1 gene deletion resulted in spermatogenic abnormalities. In addition, a significant increase in spontaneous acrosomal reaction was observed in Tas1r1 null mutant sperm whereas acrosomal secretion triggered by isolated zona pellucida or the Ca²⁺ ionophore A23187 was not different from wild-type spermatozoa. Remarkably, cytosolic Ca²⁺ levels in freshly isolated Tas1r1-deficient sperm were significantly higher compared to wild-type cells. Moreover, a significantly higher basal cAMP concentration was detected in freshly isolated Tas1r1-deficient epididymal spermatozoa, whereas upon inhibition of phosphodiesterase or sperm capacitation, the amount of cAMP was not different between both genotypes. Since Ca²⁺ and cAMP control fundamental processes during the sequential process of fertilization, we propose that the identified taste receptors and coupled signaling cascades keep sperm in a chronically quiescent state until they arrive in the vicinity of the egg - either by constitutive receptor activity and/or by tonic receptor activation by gradients of diverse chemical compounds in different compartments of the female reproductive tract.


The m-xylene biodegradation capacity of Pseudomonas putida mt-2 is submitted to adaptation to abiotic stresses: evidence from expression profiling of xyl genes.
The effect of archetypal environmental stresses on expression of the catabolic xyl genes of the TOL plasmid pWW0 of the m-xylene degrading strain Pseudomonas putida mt-2 has been investigated. To this end, a subgenomic DNA chip was employed which included structural and regulatory DNA sequences of the TOL pathway along with selected descriptors of specific physiological conditions. Cells were separately exposed to m-xylene under various oxygen tensions, temperatures and nitrogen sources as well as situations of DNA damage, oxidative stress, carbon and iron starvation, respiratory chain damage, and contact with arsenic, but at doses which did not cause a gross effect on growth or cell viability. The incidence of each stress class was categorized through the corresponding descriptors in the chip in respect to the relative output of xyl transcripts. While most of the stresses downregulated the m-xylene biodegradation-related genes, some uncouplers of the respiratory chain (azide) and small doses of arsenate appeared to stimulate their expression. The replacement of NH4+ by NO3- as N source augmented expression of the TOL cistrons also. We subsequently subjected P. putida mt-2 cells to the multiple abiotic stress brought about by exposure to crude tar from the 2002 oil spill of the Prestige tanker, which embraces a complex mixture of hydrocarbons. The resulting expression profile of xyl genes and stress-responding markers over time suggested that adaptation to external insults precedes any significant expression of the catabolic genes. The consequences of this hierarchy of responses for microbial biodegradation in situ are discussed.


Hybrid Minimally Invasive Endoscopic Approach for Gaseous Protrusion Causing L5 Radiculopathy.
We present an illustrative case of a hybrid approach between minimally invasive and endoscopic spine surgery techniques. We utilized this hybrid approach for the first 3 cases to help diminish the learning curve as we started a spinal endoscopy program. The patient is an 85-year-old woman with a prior history of arthrodesis at L3-5. She presented with severe leg pain and imaging evidence of degenerative disk disease with disk protrusion in the extraforaminal zone at L5-S1. Computed tomography imaging shows the disk protrusion to be gaseous in nature. Given failure of conservative management and patient refusal for extension of her prior fusion, she was offered a combined minimally invasive navigated technique and endoscopic approach for far lateral diskectomy. After obtaining the correct trajectory, with confirmation by intraoperative fluoroscopy, microscopic visualization was used to identify the protruded disk and the exiting nerve root (Video 1). Given the difficult visualization, a 30-degree endoscopic probe was used, which enabled real-time visualization of the gaseous protrusion being released in the liquid medium. After decompression, microscopic visualization was used for confirmation, with the patient obtaining a good surgical outcome and complete relief of her presenting pain. This case illustrates the synergism between endoscopic and minimally invasive spine surgery techniques and the unique advantages of enabling visualization of spinal anatomy through a liquid medium with the use of an endoscope.1-3.


Properties of neurons from dissociated fetal rat brain in serum-free culture.
Because of the unknown constituents and varying composition of serum, its presence in media used in cell culture unavoidably compromises attempts to study cellular mechanisms of growth and differentiation. To overcome this, we have devised a serum-free, chemically defined medium which maintains primary cultures of fetal rat brain cells for more than 6 weeks. This medium allows expression of characteristic properties of neurons and prevents overgrowth of non-neuronal elements without use of antimitotic agents. Cells prepared and plated without exposure to serum attach in less than 20 min to poly-D-lysine substratum and begin to extend processes within 1 hr. After 2 days in culture, process-bearing cells can be divided into those with characteristic neuronal morphology, including long processes which generally branch at a distance from the perikaryon, and those having the appearance of glial cells with many short, thin processes which branch frequently near the cell body. The remaining non-neuronal cells are large and flat with few or no processes. The presence of neurons and astroglia was demonstrated by immunofluorescence detection of bound tetanus toxin as a neuron-specific surface marker, and glial fibrillary acidic protein as an astroglial marker. By the 3rd day in culture, many cells of neuronal morphology were able to generate action potentials in response to electrical stimulation. The ionic composition of the inward current changes from Ca2+ to predominantly Na+ by about 10 days in culture. The presence of synaptic vesicles and myelin was demonstrated by electron microscopy. The ability of dissociated cells from mammalian brain to grow in defined medium without serum and acquire selected properties of mature cells in vivo demonstrates the potential of this culture system for neurobiological studies at the cellular level.


Variable selection under multiple imputation using the bootstrap in a prognostic study.
Missing data is a challenging problem in many prognostic studies. Multiple imputation (MI) accounts for imputation uncertainty that allows for adequate statistical testing. We developed and tested a methodology combining MI with bootstrapping techniques for studying prognostic variable selection. In our prospective cohort study we merged data from three different randomized controlled trials (RCTs) to assess prognostic variables for chronicity of low back pain. Among the outcome and prognostic variables data were missing in the range of 0 and 48.1%. We used four methods to investigate the influence of respectively sampling and imputation variation: MI only, bootstrap only, and two methods that combine MI and bootstrapping. Variables were selected based on the inclusion frequency of each prognostic variable, i.e. the proportion of times that the variable appeared in the model. The discriminative and calibrative abilities of prognostic models developed by the four methods were assessed at different inclusion levels. We found that the effect of imputation variation on the inclusion frequency was larger than the effect of sampling variation. When MI and bootstrapping were combined at the range of 0% (full model) to 90% of variable selection, bootstrap corrected c-index values of 0.70 to 0.71 and slope values of 0.64 to 0.86 were found. We recommend to account for both imputation and sampling variation in sets of missing data. The new procedure of combining MI with bootstrapping for variable selection, results in multivariable prognostic models with good performance and is therefore attractive to apply on data sets with missing values.


Genetic variability of porcine pegivirus in pigs from Europe and China and insights into tissue tropism.
Pegiviruses belong to the family Flaviviridae and have been found in humans and other mammalian species. To date eleven different pegivirus species (Pegivirus A-K) have been described. However, little is known about the tissue tropism and replication of pegiviruses. In 2016, a so far unknown porcine pegivirus (PPgV, Pegivirus K) was described and persistent infection in the host, similar to human pegivirus, was reported. In this study, qRT-PCR, phylogenetic analyses and fluorescence in situ hybridization (FISH) were implemented to detect and quantify PPgV genome content in serum samples from domestic pigs from Europe and Asia, in tissue and peripheral blood mononuclear cell (PBMC) samples and wild boar serum samples from Germany. PPgV was detectable in 2.7% of investigated domestic pigs from Europe and China (viral genome load 2.4 × 102 to 2.0 × 106 PPgV copies/ml), while all wild boar samples were tested negative. Phylogenetic analyses revealed pairwise nucleotide identities >90% among PPgVs. Finally, PPgV was detected in liver, thymus and PBMCs by qRT-PCR and FISH, suggesting liver- and lymphotropism. Taken together, this study provides first insights into the tissue tropism of PPgV and shows its distribution and genetic variability in Europe and China.


Genomic insights into cyanobacterial protein translocation systems.
Cyanobacteria are ubiquitous oxygenic photosynthetic bacteria with a versatile metabolism that is highly dependent on effective protein targeting. Protein sorting in diderm bacteria is not trivial and, in cyanobacteria, even less so due to the presence of a complex membrane system: the outer membrane, the plasma membrane and the thylakoid membrane. In cyanobacteria, protein import into the thylakoids is essential for photosynthesis, export to the periplasm fulfills a multifunctional role in maintaining cell homeostasis, and secretion mediates motility, DNA uptake and environmental interactions. Intriguingly, only one set of genes for the general secretory and the twin-arginine translocation pathways seem to be present. However, these systems have to operate in both plasma and thylakoid membranes. This raises the question of how substrates are recognized and targeted to their correct, final destination. Additional complexities arise when a protein has to be secreted across the outer membrane, where very little is known regarding the mechanisms involved. Given their ecological importance and biotechnological interest, a better understanding of protein targeting in cyanobacteria is of great value. This review will provide insights into the known knowns of protein targeting, propose hypotheses based on available genomic sequences and discuss future directions.


A CYP19 (aromatase) polymorphism is associated with increased premenopausal breast cancer risk.
Due to the established association between estrogen levels and breast cancer risk, polymorphic variation in genes regulating estrogen levels is thought to be related to breast cancer risk. Aromatase, the protein product of the CYP19 gene, is involved in the production of endogenous estrogens via androgen conversion. We examined whether polymorphic variation in CYP19 associated with increased breast cancer risk in a population based case-control study. We examined two single nucleotide polymorphisms (SNP), rs1008805 (A/G) and rs730154 (C/T), which have been shown to tag SNPs within two different haplotype blocks in CYP19. Among premenopausal women, the presence of at least one G allele at rs1008805 was significantly associated with an increase in the risk of breast cancer (OR = 1.72 [95% CI, 1.20-2.49]), especially with estrogen and progesterone receptor negative breast cancer (OR = 3.89 [1.74-8.70] and OR = 2.52 [1.26-5.05], respectively). No association was observed among postmenopausal women (OR = 1.06 [0.82-1.36]). There was no significant association between rs730154 and breast cancer, regardless of menopausal status. Our results suggest that premenopausal women carrying the G allele at CYP19 rs1008805 have increased risk of breast cancer. The finding supports the potential role of variation in estrogen biosynthesis genes in premenopausal breast cancer risk.


Characterization and IL-15 dependence of NK cells in humanized mice.
Natural Killer cells can distinguish between healthy and malignant cells and have the unique ability to lyse tumour cells without prior sensitization. Differences between murine and human NK cells complicate the translation of this knowledge into useful therapeutics. Humanized mouse models that support the development of human leukocytes are a promising avenue of research that aims to address this problem. Here we provide an in-depth phenotypic analysis of human NK cells in Balb/c Rag2(-/-)γ(c)(-/-) mice reconstituted with human hematopoietic stem cells. We have examined the development of NK cells in bone marrow, thymous, spleen, lymph node (LN) and liver. Interestingly, in naive reconstituted mice, NK cells were found in thymus and LN, but not in bone marrow. These NK cells expressed several inhibitory and activating receptors needed for malignant cell detection. Furthermore, we confirm that administration of recombinant human interleukin-15 (rhIL-15) or Ad-vector expressing hIL-15 is able to significantly enhance NK cell development and maturation, particularly in bone marrow and liver, in this model. Our results suggest that human NK cells developed in mice may have phenotypes and tissue distributions similar to those seen in human.


[Half a century of personally experienced history of psychiatry].
This report consists of the author's personal experience in German psychiatry during the second half of the last century. The focus is on scientific development in the area of biological psychiatry, a branch of psychiatry dominated, during the second quarter of the 20th century, by Kretschmer's constitutional conceptions. These, however, could not be substantiated by the author's and his co-worker's extensive research using biometrical and statistical methods still rather unusual for German psychiatry at that time. In particular, the extent as well as kind of correlation between physique and personality and the differences in physical shape between patients with either schizophrenic or affective psychoses were invalidated. The latter could largely be explained by the age dependence of physique and type of psychosis. In the psychopharmacological phase of psychiatric development following that of constitutional biology, the focus of research moved to pharmacology and, in connection with it, to neurochemistry and neuroendocrinology. In this context, objectifying and quantifying techniques, also in the area of psychopathology, became more and more important for biological research in psychiatry. This is exemplified by selected investigations of ours at the Max Planck Institute of Psychiatry in Munich. Methodologically, this prepared the ground for the neurobiological phase of development induced by progress in neuroimaging and molecular genetics which marked the transition into the 21st century.


Navigating ambivalence.
This commentary explores the sometimes uncomfortable ambivalence that colors most people's experience of disability, either as an aspect of a person's own identity, or as an aspect of a person's interactions with a person with a disability. Disabilities are experienced by many as both a positive and a negative aspect of a person's identity. The commentary describes the work of two disability scholars who have recently explored how this ambivalence affects both legal schemes and parenting decisions, and argue for a 'destigmatizing' of this ambivalence. The framework provided by these scholars illuminates ways in which ambivalence about the experience of disability is revealed by many of the contributors to this collection of essays, and suggests that an open confrontation with this ambivalence may facilitate more satisfactory relationships between people with and people without disabilities.


Halogen bond tunability I: the effects of aromatic fluorine substitution on the strengths of halogen-bonding interactions involving chlorine, bromine, and iodine.
In the past several years, halogen bonds have been shown to be relevant in crystal engineering and biomedical applications. One of the reasons for the utility of these types of noncovalent interactions in the development of, for example, pharmaceutical ligands is that their strengths and geometric properties are very tunable. That is, substitution of atoms or chemical groups in the vicinity of a halogen can have a very strong effect on the strength of the halogen bond. In this study we investigate halogen-bonding interactions involving aromatically-bound halogens (Cl, Br, and I) and a carbonyl oxygen. The properties of these halogen bonds are modulated by substitution of aromatic hydrogens with fluorines, which are very electronegative. It is found that these types of substitutions have dramatic effects on the strengths of the halogen bonds, leading to interactions that can be up to 100% stronger. Very good correlations are obtained between the interaction energies and the magnitudes of the positive electrostatic potentials (σ-holes) on the halogens. Interestingly, it is seen that the substitution of fluorines in systems containing smaller halogens results in electrostatic potentials resembling those of systems with larger halogens, with correspondingly stronger interaction energies. It is also shown that aromatic fluorine substitutions affect the optimal geometries of the halogen-bonded complexes, often as the result of secondary interactions.


Variance components in discrete force production tasks.
The study addresses the relationships between task parameters and two components of variance, "good" and "bad", during multi-finger accurate force production. The variance components are defined in the space of commands to the fingers (finger modes) and refer to variance that does ("bad") and does not ("good") affect total force. Based on an earlier study of cyclic force production, we hypothesized that speeding-up an accurate force production task would be accompanied by a drop in the regression coefficient linking the "bad" variance and force rate such that variance of the total force remains largely unaffected. We also explored changes in parameters of anticipatory synergy adjustments with speeding-up the task. The subjects produced accurate ramps of total force over different times and in different directions (force-up and force-down) while pressing with the four fingers of the right hand on individual force sensors. The two variance components were quantified, and their normalized difference was used as an index of a total force stabilizing synergy. "Good" variance scaled linearly with force magnitude and did not depend on force rate. "Bad" variance scaled linearly with force rate within each task, and the scaling coefficient did not change across tasks with different ramp times. As a result, a drop in force ramp time was associated with an increase in total force variance, unlike the results of the study of cyclic tasks. The synergy index dropped 100-200 ms prior to the first visible signs of force change. The timing and magnitude of these anticipatory synergy adjustments did not depend on the ramp time. Analysis of the data within an earlier model has shown adjustments in the variance of a timing parameter, although these adjustments were not as pronounced as in the earlier study of cyclic force production. Overall, we observed qualitative differences between the discrete and cyclic force production tasks: Speeding-up the cyclic tasks was associated with better adjustments of the timing accuracy, which helps achieve comparable force variance in tasks with different rates of force production. This does not happen in discrete tasks. The lack of scaling of the anticipatory changes in the synergy index with ramp time is the first reported feature that distinguishes anticipatory synergy adjustments from anticipatory postural adjustments. We discuss the differences between the cyclic and discrete tasks within a hierarchical control scheme offered by Schöner.


Wheat litter and feed with aluminosilicates for improved growth and meat quality in broiler chickens.
Natural minerals have many beneficial properties in poultry production, taking into account production as well as hygiene, health, safety, and quality of broiler meat. The aim of the study was to evaluate the effect of aluminosilicates in feed and litter on the growth performance and meat quality in chickens. Aluminosilicates, including halloysite and zeolite, could be a good alternative for synthetics, as a good solution for the environment in line with the current trends. Five-hundred male Ross 308 chickens were managed in 5 groups (10 replicates/group): LITTER: 1, control; 2, 0.95 kg/m2 of halloysite; 3, 0.475 kg/m2 of halloysite and 0.475 zeolite; 4, 0.95 kg/m2 of zeolite; 5, 0.25 kg/m2of halloysite and 0.7 kg/m2 of zeolite. FEED: groups 2-5, halloysite and zeolite addition (25:75 ratio; 0.5-2%). Growth performance (body weight and feed indicators), carcass, and meat quality (pH, colour, water-holding capacity, chemical composition of muscles) were recorded. The experimental setup, where the aluminosilicate additives were applied simultaneously, was proposed and approved by experts after pilot testing and on the basis of extensive literature where feed or litter additives were tested. Body weight and its gain were higher in groups 3 and 4 than in 1, and feed intake was higher in 4. The weight of the carcass and some of its components, including muscles and skin with subcutaneous fat, were higher in 2-4. Water loss from leg muscles was lower in 4. The content of protein in muscles was significantly higher in 3. The addition of aluminosilicates in feed and litter had a positive effect on the growth performance and some traits of carcasses and meat quality, especially in group 3. Halloysite and zeolite can be used in feed and litter (especially 0.475 kg/m2 for each mineral in the wheat litter).


Proximity inheritance explains the evolution of cooperation under natural selection and mutation.
In this paper, a mechanism called proximity inheritance is introduced in the birth-death process of a networked population involving the Prisoner's Dilemma game. Different from the traditional birth-death process, in the proposed model, players are distributed in a spatial space and offspring is distributed in the neighbourhood of its parents. That is, offspring inherits not only the strategy but also the proximity of its parents. In this coevolutionary game model, a cooperative neighbourhood gives more neighbouring cooperative offspring and a defective neighbourhood gives more neighbouring defective offspring, leading to positive feedback among cooperative interactions. It is shown that with the help of proximity inheritance, natural selection will favour cooperation over defection under various conditions, even in the presence of mutation. Furthermore, the coevolutionary dynamics could lead to self-organized substantial network clustering, which promotes an assortment of cooperative interactions. This study provides a new insight into the evolutionary mechanism of cooperation in the absence of social attributions such as reputation and punishment.


Dental hygiene education for nursing staff in a nursing home for older people.
This paper is a report of a study evaluating the effect of a repeated education programme for nursing staff in a home for older people. A strong relationship exists between oral infections and general health complications (especially aspiration pneumonia) among nursing home residents and hospitalized older people. Thus, nursing staff need to be educated in oral hygiene measures. Forty-three nursing home resident older people (12 men, 31 women, age range 69-99 years) were included in a dental hygiene and gingivitis evaluation using gingival bleeding scores and modified plaque scores. Evaluation was conducted before and 3 weeks after a repeated dental hygiene education for nursing staff at a nursing home in Sweden in 2008. Dental hygiene education had been given 1.5 years previously. Forty-one residents (12 men and 29 women) were available for evaluation after the repeated dental hygiene education (one died, one had had teeth extracted). There was a reduction in gingival bleeding scores (P < 0.001), and in plaque scores (P < 0.001). Repeated dental hygiene education improves the dental hygiene among nursing home resident older people. In order to succeed it may be necessary to address attitudes and perceptions towards oral care in such a dental hygiene education programme for nursing staff. Improved oral hygiene contributes to reducing the incidence of healthcare-associated pneumonia among nursing home resident older people, and thus to reduced healthcare costs.


Interferon-Beta, a Decisive Factor in Angiogenesis and Arteriogenesis.
In this review we discuss the current literature on the effects of type I interferons (IFN) and their downstream effectors on vascular growth in experimental models in vitro and in vivo. In addition to its well-documented role in angiogenesis, that is, the growth of new capillaries from existing vessels, we will also describe emerging evidence and mechanisms by which type I IFN may inhibit arteriogenesis, that is, the expansive remodeling of existing collateral arteries. Crucial in both processes is the common role of circulating monocytes, which are known to act as pivotal cellular modulators in revascularization through secreted chemokines, proteases, and growth factors. These secreted molecules, which are all modulated by IFN signaling, act via degradation of the extracellular matrix and by stimulating the proliferation of vascular smooth muscle cells and endothelial cells. Thus, next to the antiviral and immunomodulatory activities of type I IFNs, a potent role of IFN-β as modulator of revascularization is now emerging and may be considered a potential clinical target for the stimulation of angiogenesis and arteriogenesis in ill-perfused tissues.


Dose-finding study of the novel tuberculosis vaccine, MVA85A, in healthy BCG-vaccinated infants.
BCG, the only licensed tuberculosis vaccine, affords poor protection against lung tuberculosis in infants and children. A new tuberculosis vaccine, which may enhance the BCG-induced immune response, is urgently needed. We assessed the safety of and characterized the T cell response induced by 3 doses of the candidate vaccine, MVA85A, in BCG-vaccinated infants from a setting where tuberculosis is endemic. Infants aged 5-12 months were vaccinated intradermally with either 2.5 × 10(7), 5 × 10(7), or 10 × 10(7) plaque-forming units of MVA85A, or placebo. Adverse events were documented, and T-cell responses were assessed by interferon γ (IFN-γ) enzyme-linked immunospot assay and intracellular cytokine staining. The 3 MVA85A doses were well tolerated, and no vaccine-related serious adverse events were recorded. MVA85A induced potent, durable T-cell responses, which exceeded prevaccination responses up to 168 d after vaccination. No dose-related differences in response magnitude were observed. Multiple CD4 T cell subsets were induced; polyfunctional CD4 T cells co-expressing T-helper cell 1 cytokines with or without granulocyte-macrophage colony-stimulating factor predominated. IFN-γ-expressing CD8 T cells, which peaked later than CD4 T cells, were also detectable. MVA85A was safe and induced robust, polyfunctional, durable CD4 and CD8 T-cell responses in infants. These data support efficacy evaluation of MVA85A to prevent tuberculosis in infancy. Clinical Trials Registration. NCT00679159.


[Prevalence in adults of attention deficit hyperactivity disorder using the medical records of primary care].
ADHD in adults is not uncommon and, according to recent epidemiological data, has a population prevalence of 3-4%. However, there is major unawareness of this disorder among doctors, particularly in primary care. The aim of this study is to determine the prevalence of the diagnosis of ADHD recorded in adults and the proportion of patients with drug prescriptions for this disorder in primary care. This is a cross-sectional study on databases of computerised primary care medical records. The target population is adults (18-44 years) assigned to primary care centres of the Catalan Health Institute (n = 2,452,107). We obtained the proportion of patients with ADHD (F90/ICD- 10 code) registered as active problem, and the proportion of patients with a specific prescription for ADHD in adults: methylphenidate, methylphenidate extended release or atomoxetine. The prevalence of recorded ADHD is 0.04% (0.07% in men, 0.02% in women). The percentage of patients with a specific prescription for ADHD is 0.07% (0.08% in men, 0.05% in women). 32.05% of ADHD patients had specific prescription. The diagnosis of ADHD in adults and specific treatment are extremely low in primary care. These results contrast with population data: registered ADHD is 1/85 of the population prevalence.


Eurasian Red Squirrels Show Little Seasonal Variation in Metabolism in Food-Enriched Habitat.
Energy expenditure and ambient temperature (Ta) are intrinsically linked through changes in an animal's metabolic rate. While the nature of this relationship is stable, the breadth of change in thermoregulatory cost varies with body size and physiological acclimatization to season. To explore seasonal metabolic changes of small mammals, we studied a population of Eurasian red squirrels (Sciurus vulgaris) in a seminatural environment with a year-round supply of natural and supplemented food. In each season we measured the metabolic rate of wild-caught red squirrels, using open-flow respirometry, and hypothesized that individuals would make adjustments to contend with seasonal weather conditions. In comparison to summer animals, we predicted that winter squirrels would show (1) an increase in metabolic rate within the thermoneutral zone, (2) a decrease in the lower critical temperature of the thermoneutral zone, (3) a shallower slope of resting metabolic rate with decreasing Ta, and (4) lower thermal conductance. Surprisingly, we observed only minor changes in resting metabolic rate, and energetic modeling suggested that the scope of change was unlikely to be of ecological consequence. Hair area density was higher in winter than in summer, corresponding to a slightly elevated thermal conductance in summer, while body mass was reasonably constant year-round. We conclude that the scope of physiological seasonal adaptation is minimal when food is abundant and that squirrels instead rely on adjustments in activity to reduce exposure to low Ta. We suggest that this may explain the squirrel's success in a wide range of habitats, including urban areas, which require a rapid and flexible response to environmental changes and may indicate the capacity of other small mammal species to cope with environmental disturbance.


Genetics of bicuspid aortic valve aortopathy.
The incidence of aortic dilation and acute complications (rupture and dissection) is higher in patients with a bicuspid aortic valve (BAV), the most frequent congenital heart defect.The present review focuses on the current knowledge in the genetics of BAV, emphasizing the clinical implications for early detection and personalized care. BAV is a highly heritable trait, but the genetic causes remain largely elusive. NOTCH1 is the only proven candidate gene to be associated with both familial and sporadic BAV. Other genes have been reported to be associated with BAV, but some of these associations may result from coexisting disease.The application of modern high-throughput technologies (next generation sequencing, genome-wide copy number and genome-wide methylation arrays) have begun to dissect the genetic heterogeneity underlying BAV as well as the diverse molecular pathways involved in the progression of BAV aortopathy. The clinical variability seen in BAV aortopathy, in terms of phenotype and natural/clinical history, suggests complex interactions between primary genetic defects, other modifier genes, epigenetic factors (DNA methylation or histone modifications, microRNA) and environmental factors (disturbed flow). Integrated, more comprehensive studies are needed for elucidating these connections to develop more individualized and accurate risk assessment methods.


Metabolic cooperation and spatiotemporal niche partitioning in a kefir microbial community.
Microbial communities often undergo intricate compositional changes yet also maintain stable coexistence of diverse species. The mechanisms underlying long-term coexistence remain unclear as system-wide studies have been largely limited to engineered communities, ex situ adapted cultures or synthetic assemblies. Here, we show how kefir, a natural milk-fermenting community of prokaryotes (predominantly lactic and acetic acid bacteria) and yeasts (family Saccharomycetaceae), realizes stable coexistence through spatiotemporal orchestration of species and metabolite dynamics. During milk fermentation, kefir grains (a polysaccharide matrix synthesized by kefir microorganisms) grow in mass but remain unchanged in composition. In contrast, the milk is colonized in a sequential manner in which early members open the niche for the followers by making available metabolites such as amino acids and lactate. Through metabolomics, transcriptomics and large-scale mapping of inter-species interactions, we show how microorganisms poorly suited for milk survive in-and even dominate-the community, through metabolic cooperation and uneven partitioning between grain and milk. Overall, our findings reveal how inter-species interactions partitioned in space and time lead to stable coexistence.


[Characteristics of pharmacological and toxic effects of verapamil during cardiac arrhythmia in thyrotoxic and hypothyroid rats].
Aim of this study was to investigate antiarrhythmic and toxic effects of verapamil in mice and rats with thyrotoxicosis and hypothyroidism. We found that changes of thyroid status lead to the increased sensitivity of animals to toxicity of verapamil. At single intraperitoneal introduction of verapamil, LD50 was 118 +/- 7,7 mg/kg for control euthyroid mice, 53 +/- 4,1 mg/kg for hypothyroid mice, and 77 +/- 6,5 mg/kg for thyrotoxic mice. Sensitivity of rat myocardium to arrhythmogenic effects of calcium chloride increased with development of thyrotoxicosis and hypothyroidism. Effective arrhythmogenic dose of CaCl2 was 200 mg/kg for euthyroid rats and 140 mg/kg for rats with thyroid dysfunction. Intravenous introduction of verapamil had antiarrhythmic activity in rats with experimental thyroid dysfunction but at a lower effective dose. Effective preventive dose of verapamil was 3 - 4,5 mg/kg for euthyroid rats and 2 - 2,5 mg/kg for rats with thyrotoxicosis and hypothyroidism. Effective dose of verapamil during rhythm disturbance was 3 mg/kg for control euthyroid rats and 1,5 - 2 mg/kg for rats with abnormal thyroid status. These results provide a basis for the new individual approach for treatment of patients with cardiac arrhythmia combined with thyroid dysfunction.


Incubation conditions are more important in determining early thermoregulatory ability than posthatch resource conditions in a precocial bird.
Recent research in birds suggests that investing in incubation is one mechanism by which parents can enhance the phenotype of their offspring. Posthatch environmental conditions can also shape an individual's phenotype, and it is thus possible for pre- and posthatch conditions to have interactive effects on an individual's phenotype. In this study, we examined the individual and interactive effects of prehatch incubation temperature and posthatch food availability on growth, food consumption, and thermoregulatory ability in wood duck (Aix sponsa) ducklings. Eggs were incubated at one of three temperatures (35.0°, 35.9°, or 37.0°C), and then ducklings were reared on an either ad lib. or time-restricted diet for 12 d after hatching. We found that food availability influenced duckling growth, with the slowest growth occurring in ducklings fed the restricted diet. Incubation temperature also interacted with food conditions to influence duckling growth: ducklings fed ad lib. from the lowest incubation temperature grew slower than ducklings fed ad lib. from the higher incubation temperatures. Most importantly, we found that the improvement in a duckling's ability to maintain body temperature in the face of a thermal challenge was influenced by embryonic incubation temperature but not feeding conditions. Ducklings from the highest incubation temperature experienced the greatest improvement in thermoregulatory performance with age. Our findings suggest that the prehatch environment is more important than posthatch resource conditions in determining some physiological functions and underscores the important role that incubation temperature plays in determining offspring phenotype in birds.


Lenalidomide can induce long-term responses in patients with multiple myeloma relapsing after multiple chemotherapy lines, in particular after allogeneic transplant.
Evidence of long-term response to lenalidomide in heavily pretreated patients with multiple myeloma is lacking. This study sought to assess whether long-term responders exist, long-term responders' characteristics, and predictive factors of a long-term response. One hundred and four patients with multiple myeloma treated with lenalidomide and dexamethasone after ≥2 therapy lines (median, 3) were analyzed. Long-term response was defined as at least a partial response (≥PR) lasting ≥12 months. The overall response rate was 73%, and 80.3% of the responses were achieved within 5 months. The median response was 14.3 months. Patients evaluable for long-term response numbered 87, and a total of 47% were long-term responders. Compared to non-long-term responders, long-term responders had better overall survival, less light-chain multiple myeloma, and higher incidence of t(11;14). Previous allogeneic transplant (alloSCT) and the response quality predicted a long-term response. In conclusion, patients treated with lenalidomide can become long-term responders; alloSCT and response quality predict long-term response.


[Clinical and pathogenetic features of occupational bronchitis].
With multidimentional correlation analysis, the authors studied clinical and pathogenetic features of occupational bronchitis development. Analysis covered relationships of symptoms and syndromes of the disease, main spirometric signs of bronchial obstruction, presence and character of exogenic risk factors and the patients age. The authors revealed regularities of interrelations of clinical, instrumental and environmental characteristics of dust diseases, their role in unfavorable course and outcomes of occupational bronchitis.


Interstitial fluid drainage is impaired in ischemic stroke and Alzheimer's disease mouse models.
The interstitial fluid (ISF) drainage pathway has been hypothesized to underlie the clearance of solutes and metabolites from the brain. Previous work has implicated the perivascular spaces along arteries as the likely route for ISF clearance; however, it has never been demonstrated directly. The accumulation of amyloid β (Aβ) peptides in brain parenchyma is one of the pathological hallmarks of Alzheimer disease (AD), and it is likely related to an imbalance between production and clearance of the peptide. Aβ drainage along perivascular spaces has been postulated to be one of the mechanisms that mediate the peptide clearance from the brain. We therefore devised a novel method to visualize solute clearance in real time in the living mouse brain using laser guided bolus dye injections and multiphoton imaging. This methodology allows high spatial and temporal resolution and revealed the kinetics of ISF clearance. We found that the ISF drains along perivascular spaces of arteries and capillaries but not veins, and its clearance exhibits a bi-exponential profile. ISF drainage requires a functional vasculature, as solute clearance decreased when perfusion was impaired. In addition, reduced solute clearance was observed in transgenic mice with significant vascular amyloid deposition; we suggest the existence of a feed-forward mechanism, by which amyloid deposition promotes further amyloid deposition. This important finding provides a mechanistic link between cerebrovascular disease and Alzheimer disease and suggests that facilitation of Aβ clearance along the perivascular pathway should be considered as a new target for therapeutic approaches to Alzheimer disease and cerebral amyloid angiopathy.


Preoperative COVID-19 Testing for Elective Ophthalmological Procedure in a Tertiary Health Care Centre: Our Experience During the Pandemic.
To study the prevalence of coronavirus infection among asymptomatic patients requiring eye surgery and the role of screening in prevention of spread of infection among the healthcare workers. The prospective observational study was conducted in the Ophthalmology department of a tertiary care center in Delhi from September 2020 to December 2020. Patients requiring elective ophthalmological procedures in a tertiary care hospital were screened for coronavirus using the RT-PCR method. Testing methods and results were documented. Among the 218 asymptomatic patients posted for elective surgery in that period, 16 (7.3%) were found to be positive for COVID-19. Those who tested positive were advised home isolation and surgery was postponed for the next 14 days. No complications were reported in these patients. One health-care worker also tested positive for COVID-19 during our study period. In our study, 1 out of 14 asymptomatic patients were found to be carriers for the novel virus. Asymptomatic COVID-infected patients may lead to transmission of the virus inside the hospital among the visiting patients and hospital staff while they have no adverse effect on the surgery and its outcome.


Age-related decreases in GTP-cyclohydrolase-I immunoreactive neurons in the monkey and human substantia nigra.
Guanosine triphosphate cyclohydrolase I (GTPCHI) is a critical enzyme in catecholamine function and is rate limiting for the synthesis of the catecholamine co-factor tetrahydrobiopterin. The present study assessed the distribution of GTPCHI immunoreactivity (-ir) within the monkey and human ventral midbrain and determined whether its expression is altered as a function of age. Light and confocal microscopic analyses revealed that young monkeys and humans displayed GTPCHI-ir within melanin-containing and tyrosine-hydroxylase-ir neurons in primate substantia nigra. Stereological counts revealed that there was a 67.4% reduction in GTPCHI-ir neuronal number, a 63.5% reduction in GTPCHI-ir neuronal density, and a 37.6% reduction in neuronal volume in aged monkeys relative to young cohorts. Similar age-related changes were seen in humans, in whom there were significant reductions in the number of GTPCHI-ir nigral neurons in middle age (58.4%) and aged (81.5%) cases relative to young cohorts. The density of GTPCHI-ir neurons within the nigra was similarly reduced in middle-aged (63.0%) and aged (81.8%) cases. In contrast to monkeys, aged humans did not display shrinkage in the volume of GTPCHI-ir nigral neurons. The presence of numerous melanin-positive, but GTPCHI-ir immunonegative, neurons in the aged monkey and human nigra indicates that these decreases represent an age-related phenotypic downregulation of this enzyme and not a loss of neurons per se. These data indicate that there is a dramatic decrease in GTPCHI-ir in nonhuman primates and humans as a function of age and that loss of this enzyme may be partly responsible for the age-related decrease in dopaminergic tone within nigrostriatal systems.


Fatty acid oxidation is directly regulated by carbohydrate metabolism during exercise.
We determined whether increased glycolytic flux from hyperglycemia and hyperinsulinemia directly reduces fatty acid oxidation during exercise. Fatty acid oxidation rates were measured during constant-rate intravenous infusion of trace amounts of a long-chain fatty acid ([1-13C]palmitate; Pal) vs. a medium-chain fatty acid ([1-13C]octanoate; Oct). Six endurance-trained men cycled for 40 min at 50% of maximal O2 uptake 1) after an overnight fast ("fasting") and 2) after ingestion of 1.4 g/kg of glucose at 60 min and again 10 min before exercise (Glc). Glc caused hyperinsulinemia, a preexercise blood glucose of 6 mM, and a 34% reduction in total fat oxidation during exercise due to an approximately equal reduction in oxidation of plasma-free fatty acids (FFA) and intramuscular triglycerides (all P < 0.05). Oxidation of Pal was significantly reduced during Glc compared with fast (i.e., 70.0 +/- 4.1 vs. 86.0 +/- 1.9% of tracer infusion rate; P < 0.05). However, Glc had no effect on Oct oxidation, which is apparently not limited by mitochondrial transport. Furthermore, Glc reduced plasma FFA appearance 36% (P < 0.05), indicating a coordination of effects on adipose tissue and muscle. In summary, substrate oxidation during exercise can be regulated by increased glycolytic flux that is accompanied by a direct inhibition of long-chain fatty acid oxidation. These observations indicate that carbohydrate availability can directly regulate fat oxidation during exercise.


N1-guanyl-1,7-diaminoheptane enhances the chemosensitivity of NSCLC cells to cetuximab through inhibition of eukaryotic translation initiation factor 5A2 activation.
N1-guanyl-1, 7-diaminoheptane (GC7), an inhibitor of deoxyhypusine synthase has been shown to exhibit significant anti-cancer activity. However, the biological role of eukaryotic translation initiation factor 5A2 activation (EIF5A2) and GC7 on drug resistance in non-small cell lung cancer (NSCLC) has not been investigated. In this study, we aimed to investigate the therapeutic effect of GC7 combined with cetuximab in NSCLC therapy. The current study used cell viability assays, EdU incorporation assays, and western blot to detect that the GC7 exhibited synergistic cytotoxicity with cetuximab in NSCLC. CCK-8 assays showed that combined treatment with GC7 and cetuximab significantly inhibited the viabilities in three NSCLC cell lines. In addition, EdU incorporation assays also indicated that GC7 co-treatment remarkably enhanced the cetuximab sensitivity in NSCLC cells. Nevertheless, down-regulation of EIF5A2 diminished the regulatory role of GC7 in cetuximab cytotoxicity. Western blot showed that transfection of EIF5A2 siRNA significantly suppressed the protein expression of EIF5A2 in NSCLC cells. These findings demonstrate that combined treatment with GC7 could enhance cetuximab sensitivity by inhibiting EIF5A2 in NSCLC cells, implying the potential clinical application of GC7 in cetuximab-based chemotherapy for NSCLC patients.


High-Quality Genome Sequence of the Highly Resistant Bacterium Staphylococcus haemolyticus, Isolated from a Neonatal Bloodstream Infection.
Using Illumina HiSeq and PacBio technologies, we sequenced the genome of the multidrug-resistant bacterium Staphylococcus haemolyticus, originating from a bloodstream infection in a neonate. The sequence data can be used as an accurate reference sequence.


Molecular Mechanisms Underlying the Link between Diet and DNA Methylation.
DNA methylation is a vital modification process in the control of genetic information, which contributes to the epigenetics by regulating gene expression without changing the DNA sequence. Abnormal DNA methylation-both hypomethylation and hypermethylation-has been associated with improper gene expression, leading to several disorders. Two types of risk factors can alter the epigenetic regulation of methylation pathways: genetic factors and modifiable factors. Nutrition is one of the strongest modifiable factors, which plays a direct role in DNA methylation pathways. Large numbers of studies have investigated the effects of nutrition on DNA methylation pathways, but relatively few have focused on the biochemical mechanisms. Understanding the biological mechanisms is essential for clarifying how nutrients function in epigenetics. It is believed that nutrition affects the epigenetic regulations of DNA methylation in several possible epigenetic pathways: mainly, by altering the substrates and cofactors that are necessary for proper DNA methylation; additionally, by changing the activity of enzymes regulating the one-carbon cycle; and, lastly, through there being an epigenetic role in several possible mechanisms related to DNA demethylation activity. The aim of this article is to review the potential underlying biochemical mechanisms that are related to diet modifications in DNA methylation and demethylation.


Variability in exercise physiology: Can capturing intra-individual variation help better understand true inter-individual responses?
Exploring individual responses to exercise training is a growing area of interest. Understanding reasons behind true observed inter-individual responses may help personalise exercise training to maximise the benefits received. While numerous factors have been explored, an often underappreciated consideration in the sport and exercise science field is the influence intra-individual variation, both in a single measurement and in response to an intervention, may have on training outcomes. Several study designs and statistical approaches are available to incorporate intra-individual variation into interventions and accordingly provide information on whether 'true' inter-individual responses are present or if they are an artefact of intra-individual variation. However, such approaches are sparingly applied. Moreover, intra-individual variation may also be important when true inter-individual response differences are present. In this perspective piece, the concept of intra-individual variation is described before briefly summarising study designs and statistical practices to account for intra-individual variation. We then outline two examples of physiological practices (stratified randomisation and prescribing exercise programmes upon training parameters) to demonstrate why sport and exercise scientists should acknowledge intra-individual variation prior to the implementation of an intervention, which potentially offers an additional explanation behind observed true inter-individual responses to training. Repeated testing pre-implementation of exercise training would conceptually provide more confident estimates of training parameters, which if utilised in a study design will help attenuate biases that may dictate inter-individual differences. Moreover, the incorporation of intra-individual differences will facilitate insights into alternative factors that may predict and/or explain true observed individual responses to an exercise training programme.


A universal fluorometric assay strategy for glycosidases based on functional carbon quantum dots: β-galactosidase activity detection in vitro and in living cells.
The development of highly sensitive assays for glycosidases is of critical significance to understand their functions, facilely detect associated diseases and screen potential new drugs. In this work, we develop a universal assay strategy for glycosidase enzymes and inhibitor screening based on functional carbon quantum dots through a combined host-guest recognition and specific static quenching-induced signal transduction mechanism. This detection strategy is established in terms of the following facts: (1) β-cyclodextrin as a perfect host can selectively associate with p-nitrophenol due to its hydrophobic character and right size match of the cavity, which renders specific binding between β-cyclodextrin and p-nitrophenol via a host-guest recognition. (2) The formation of an inclusion complex between β-cyclodextrin modified carbon quantum dots (β-CD-CQDs) and p-nitrophenol results in fluorescence quenching with a high quenching efficiency due to the static quenching mechanism. Glycoconjugates of p-nitrophenol as the substrates could be rapidly hydrolyzed to corresponding glycose and p-nitrophenol in the presence of specific glycosidase, and the resulting p-nitrophenol induces the following host-guest interaction and static quenching leading to a change in the fluorescence signal. The activity of different glycosidase enzymes could be evaluated in the same way as long as the glycosyl unit of glycosylated substrates was changed. Here we take β-galactosidase as an example to demonstrate the applicability of the proposed detection strategy because it can act as a molecular target for primary ovarian cancers. A highly sensitive assay for β-galactosidase activity in terms of linear correlation of the fluorescence change with the β-galactosidase level was established with a low detection limit of 0.6 U L-1. Its function of inhibitor screening was also assessed by using d-galactal as the inhibitor for β-galactosidase, and the positive results indicated its feasibility to screen potential inhibitors. It is also illustrated that the nanoprobe possesses excellent biocompatibility, and can sensitively monitor the intracellular β-galactosidase level in ovarian cancer cells. This work provides a general detection method for glycosidase activity, demonstrates its applicability of monitoring the enzyme level in living cells, and broadens fluorogenic probes in fluorescence-guided diagnostics.


Emotional intelligence of mental health nurses.
The aim of this study is to gain insight into the level of emotional intelligence of mental health nurses in the Netherlands. The focus in research on emotional intelligence to date has been on a variety of professionals. However, little is known about emotional intelligence in mental health nurses. The emotional intelligence of 98 Dutch nurses caring for psychiatric patients is reported. Data were collected with the Bar-On Emotional Quotient Inventory within a cross-sectional research design. The mean level of emotional intelligence of this sample of professionals is statistically significant higher than the emotional intelligence of the general population. Female nurses score significantly higher than men on the subscales Empathy, Social Responsibility, Interpersonal Relationship, Emotional Self-awareness, Self-Actualisation and Assertiveness. No correlations are found between years of experience and age on the one hand and emotional intelligence on the other hand. The results of this study show that nurses in psychiatric care indeed score above average in the emotional intelligence required to cope with the amount of emotional labour involved in daily mental health practice. The ascertained large range in emotional intelligence scores among the mental health nurses challenges us to investigate possible implications which higher or lower emotional intelligence levels may have on the quality of care. For instance, a possible relation between the level of emotional intelligence and the quality of the therapeutic nurse-patient relationship or the relation between the level of emotional intelligence and the manner of coping with situations characterised by a great amount of emotional labour (such as caring for patients who self-harm or are suicidal).


Blood pressure visit-to-visit variability and outcomes in patients with heart failure with preserved ejection fraction.
Previous studies report that blood pressure (BP) variability is associated with increased risk of adverse outcomes in patients diagnosed with cardiovascular disease. However, studies have not fully explored this association in patients with heart failure with preserved ejection fraction (HFpEF). This study sought to explore the association between visit-to-visit variability (VVV) of BP and clinical outcomes in patients with HFpEF. A total of 1988 patients (mean age of 67.73 ± 9.22, 51.7% female) from the Treatment of Preserved Cardiac Function Heart Failure with an Aldosterone Antagonist (TOPCAT) trial were included in this study. BP-VVV was determined by standard deviation (SD) of mean systolic BP (SBP-SD) from six measurements (baseline and months 1, 2, 4, 8, and 12) during the first 12 months after randomization. Mean on-treatment SBP during the first 12 months was 127.77 ± 10.42 mmHg, and the median of SBP-SD was 8.15 mmHg. A total of 192 (9.7%) patients met the primary outcome during the subsequent median follow-up of 35.16 months, including a composite of cardiovascular death, heart failure hospitalization, or aborted cardiac arrest. Multiple Cox regression analysis showed that SBP-SD was independently associated with the increased risk of the primary outcome after adjusting for age, gender, method of BP measurement, treatment, renal function and common co-morbidities, and the mean SBP during the first 12 months [hazard ratio (HR) for fourth vs. first quartile, 1.63; 95% confidence interval (CI), 1.07-2.49; P = 0.024]. Analysis showed that SBP-SD as continuous variable was associated with a 23% increase in the risk of primary outcome (HR 1.23, 95% CI 1.06-1.43; P = 0.006). The findings of the current study show that high SBP-VVV in patients with HFpEF is associated with an increased risk of adverse outcomes independent of the mean on-treatment SBP.


The production and activity in vivo of Proteus mirabilis IgA protease in infections of the urinary tract.
Immunoblotting of urine from 21 patients of both sexes and of wide age range who had a Proteus mirabilis urinary tract infection (UTI) showed that 14 (64%) specimens contained immunoglobulin A (IgA). In nine (64%) of these the IgA heavy chain had been degraded to fragments of a size identical to those formed when purified IgA was degraded by pure P. mirabilis protease. Urine from patients with clinical evidence of upper UTI contained fragmented IgA and in some of these urine samples P. mirabilis protease activity was detectable. Urine infected with a non-proteolytic strain contained only intact IgA. It is concluded that P. mirabilis IgA protease is produced and is active during infections of the urinary tract.


A cytogenetic follow-up of some highly irradiated victims of the Chernobyl accident.
A follow-up of 10 highly irradiated men, mostly reactor crew, from the Chernobyl accident is described. Their pre-accident medical conditions and relevant medical status approximately 10-13 y later are listed. A comparison is made between estimates of their average whole-body penetrating radiation doses derived from several biological parameters. First estimates were based on their presenting severity of prodromal sickness, early changes in blood cell counts and dicentric chromosome aberrations in lymphocytes. In three cases ESR measurements on tooth enamel were also made. Retrospective dosimetry using FISH translocations was attempted 10-13 y later. This showed good agreement for those patients with the lower earlier dose estimates, up to about 3 Gy. For the others, extending up to about 12 Gy, the translocations indicated lower values, suggesting that in these cases translocations had somewhat declined. Repeated chromosomal examinations during the follow-up period showed an expected decline in dicentric frequencies. The pattern of decline was bi-phasic with a more rapid first phase, with a half-life of approximately 4 months followed by a slower decline with half-lives around 2-4 y. The rapid phase persisted for a longer time in those patients who had received the highest doses. 10-13 y later dicentric levels were still above normal background, but well below the translocation frequencies.


Subjective but Not Actigraphy-Defined Sleep Predicts Next-Day Fatigue in Chronic Fatigue Syndrome: A Prospective Daily Diary Study.
This study aimed to (1) examine the relationship between subjective and actigraphy-defined sleep, and next-day fatigue in chronic fatigue syndrome (CFS); and (2) investigate the potential mediating role of negative mood on this relationship. We also sought to examine the effect of presleep arousal on perceptions of sleep. Twenty-seven adults meeting the Oxford criteria for CFS and self-identifying as experiencing sleep difficulties were recruited to take part in a prospective daily diary study, enabling symptom capture in real time over a 6-day period. A paper diary was used to record nightly subjective sleep and presleep arousal. Mood and fatigue symptoms were rated four times each day. Actigraphy was employed to provide objective estimations of sleep duration and continuity. Multilevel modelling revealed that subjective sleep variables, namely sleep quality, efficiency, and perceiving sleep to be unrefreshing, predicted following-day fatigue levels, with poorer subjective sleep related to increased fatigue. Lower subjective sleep efficiency and perceiving sleep as unrefreshing predicted reduced variance in fatigue across the following day. Negative mood on waking partially mediated these relationships. Increased presleep cognitive and somatic arousal predicted self-reported poor sleep. Actigraphy-defined sleep, however, was not found to predict following-day fatigue. For the first time we show that nightly subjective sleep predicts next-day fatigue in CFS and identify important factors driving this relationship. Our data suggest that sleep specific interventions, targeting presleep arousal, perceptions of sleep and negative mood on waking, may improve fatigue in CFS.


A prospective evaluation of antithyroid antibody prevalence in 100 patients with systemic lupus erythematosus.
During a 6-month period, 100 patients with systemic lupus erythematosus (SLE) were consecutively studied for the presence of antithyroid antibodies and thyroid disease. Overall, the prevalence of antithyroid antibodies was similar in patients with SLE (21%) and controls (16%). However, antithyroglobulin antibodies were found in 11% of patients with SLE and only 2% of controls (p = 0.009). The levels of antimicrosomal antibodies were also different (median levels: SLE = 400; controls = 100) but this difference did not reach statistical significance. We found a significant correlation between thyroid stimulating hormone (TSH) levels and the levels of both the antithyroglobulin dilutions (p less than 0.05) and the antimicrosomal dilutions (p less than 0.05). This was not seen in the controls. A higher frequency of clinical thyroid disease was seen in patients with SLE with thyroid antibodies (5/21; 3 hypothyroid, 2 hyperthyroid) than in those without these antibodies (1/79; p = 0.001). Levels of antithyroid antibodies correlated with clinical or subclinical (marked by elevations of TSH) thyroid disease. Patients with SLE with these antibodies were significantly older (mean age 47.5 +/- 13 years) than those without antithyroid antibodies (mean age 37.5 +/- 12 years; p less than 0.001). Antithyroid antibodies define a subset of older patients with SLE with increased prevalence of both clinical and subclinical thyroid disease.


Dorsal cortical regions subserving visually guided saccades in humans: an fMRI study.
Neurophysiological studies in non-human primates have identified saccade-related neuronal activity in cortical regions including frontal (FEF), supplementary (SEF) and parietal eye fields. Lesion and neuroimaging studies suggest a generally homologous mapping of the oculomotor system in humans; however, a detailed mapping of the precise anatomical location of these functional regions has not yet been achieved. We investigated dorsal frontal and parietal cortex during a saccade task vs. central fixation in 10 adult subjects using functional magnetic resonance imaging (fMRI). The FEF were restricted to the precentral sulcus, and did not extend anteriorly into Brodmann area 8, which has traditionally been viewed as their location in humans. The SEF were located in cortex along the interhemispheric fissure and extended minimally onto the dorsal cortical surface. Parietal activation was seen in precuneus and along the intraparietal sulcus, extending into both superior and inferior parietal lobules. These findings localize areas in frontal and parietal cortex involved in saccade generation in humans, and indicate significant differences from the macaque monkey in both frontal and parietal cortex. These differences may have functional implications for the roles these areas play in visuomotor processes.


The safety and efficacy of the 5-HT 1F receptor agonist lasmiditan in the acute treatment of migraine.
Migraine is among the most disabling disorders worldwide, with a significant therapeutic need. Triptans are drugs of choice in the acute attack treatment, but they are contraindicated in patients with vascular conditions due to their potential vasoconstrictive properties. Further limitations include side effects, inconsistency in therapeutic action and possible non-response. Lasmiditan, a highly selective 5-HT1F receptor agonist, is a novel acute anti-migraine substance devoid of vasoconstriction. Areas covered: This article reviews the clinical efficacy and safety of oral and intravenous lasmiditan as a possible acute migraine treatment. We analyze all currently available results in Phase I to III studies. Expert opinion: Lasmiditan is a promising acute migraine therapy, in particular for patients at cardiovascular risk. Phase II and the first Phase III clinical trials show a significant better headache response in comparison to placebo. The efficacy of lasmiditan proves that vasoconstriction is not essential for acute migraine therapy and thereby points, in addition to a well-established trigeminal contribution, to central neuronal mechanisms in migraine pathophysiology. Lasmiditan penetrates the blood-brain barrier and CNS associated adverse events are common, but mostly in mild to moderate severity. The results of long-term Phase III studies will determine if these adverse events represent a limitation in clinical practice.


Genome-wide analysis of ZmDREB genes and their association with natural variation in drought tolerance at seedling stage of Zea mays L.
The worldwide production of maize (Zea mays L.) is frequently impacted by water scarcity and as a result, increased drought tolerance is a priority target in maize breeding programs. While DREB transcription factors have been demonstrated to play a central role in desiccation tolerance, whether or not natural sequence variations in these genes are associated with the phenotypic variability of this trait is largely unknown. In the present study, eighteen ZmDREB genes present in the maize B73 genome were cloned and systematically analyzed to determine their phylogenetic relationship, synteny with rice, maize and sorghum genomes; pattern of drought-responsive gene expression, and protein transactivation activity. Importantly, the association between the nucleic acid variation of each ZmDREB gene with drought tolerance was evaluated using a diverse population of maize consisting of 368 varieties from tropical and temperate regions. A significant association between the genetic variation of ZmDREB2.7 and drought tolerance at seedling stage was identified. Further analysis found that the DNA polymorphisms in the promoter region of ZmDREB2.7, but not the protein coding region itself, was associated with different levels of drought tolerance among maize varieties, likely due to distinct patterns of gene expression in response to drought stress. In vitro, protein-DNA binding assay demonstrated that ZmDREB2.7 protein could specifically interact with the target DNA sequences. The transgenic Arabidopsis overexpressing ZmDREB2.7 displayed enhanced tolerance to drought stress. Moreover, a favorable allele of ZmDREB2.7, identified in the drought-tolerant maize varieties, was effective in imparting plant tolerance to drought stress. Based upon these findings, we conclude that natural variation in the promoter of ZmDREB2.7 contributes to maize drought tolerance, and that the gene and its favorable allele may be an important genetic resource for the genetic improvement of drought tolerance in maize.


[The national history space chronobiology].
Analyzed were theories stated in the treatises of Russian researchers V.I. Nikolsky, N.Ya. Perna and A.L. Chizhevsky dedicated to the place of the phenomenon of rhythm in biology and medicine. The authors underline the inceptive role of the fundamental ideas of these scientists in formation and development of space biorhythmology in this country. Presented are main postulates in the area of space biorhythmology formulated in the period of fulfillment of the Russian piloted space programmes.


Metrologies for the phase characterization of attosecond extreme ultraviolet optics.
Extreme ultraviolet (EUV) optics play a key role in attosecond science since only with higher photon energies is it possible to achieve the wide spectral bandwidth required for ultrashort pulses. Multilayer EUV mirrors have been proposed and are being developed to temporally shape (compress) attosecond pulses. To fully characterize a multilayer optic for pulse applications requires not only knowledge of the reflectivity, as a function of photon energy, but also the reflected phase of the mirror. We develop the metrologies to determine the reflected phase of an EUV multilayer mirror using the photoelectric effect. The proposed method allows one to determine the optic's impulse response and hence its pulse characteristics.


Electro-acupuncture in the treatment of children with monosymptomatic nocturnal enuresis.
The aim of this study was to investigate the effects of a long series of electro-acupuncture (EAP) sessions on bedwetting symptoms. Twenty-five children (age range 7-16 years) with monosymptomatic nocturnal enuresis and treated earlier without success were included in the study. The median number of wet nights per week was 4.7 before treatment. Bedwetting, voided volume, sleep and nocturia were evaluated 3 weeks, 3 months and 6 months after 20 sessions of EAP lasting 8 weeks. All the children, with the exception of one, tolerated EAP treatment well. At the three follow-up sessions it was found that the number of dry nights had increased gradually from a median of 2.3 in the pre-test to 3.0, 4.3 and 5.0 per week, respectively. Compared to pre-treatment findings there were more dry nights in 65% of the children (p < 0.001) and 5 out of 23 children were responders (> 90% reduction of the numbers of wet nights) at the 6 months' follow-up. According to the parents, the sleep arousal threshold had decreased in about 50% of the children.


[Osteoprotegerin (OPG/OCIF) inhibits bone destruction in rheumatoid arthritis models].
Recent discovery of factors involved in bone destruction in Rheumatoid Arthritis (RA) identified its molecular mechanism. Osteoclast differentiation factor (ODF, also called receptor activator of NF-kappaB ligand (RANKL) ) that controls osteoclast differentiation and function has a major role in the bone destruction among them. Osteoclastogenesis inhibitory factor (OCIF, also called osteoprotegerin (OPG) ) that is a decoy receptor for ODF/RANKL is a specific inhibitor of bone destruction. OPG/OCIF may be useful for and applicable to the treatment of bone destruction in RA.


Clonal structure through space and time: High stability in the holothurian Stichopus chloronotus (Echinodermata).
Sea cucumbers are increasingly exploited for human consumption and for their curative properties, and many wild populations are now depleted or in danger of extinction. While aquaculture is seen as an alternative to fisheries and as a mean to restore wild populations, more knowledge is needed on their reproductive strategies to render this practice efficient, notably for fissiparous holothurians, which are some of the mobile animals able of asexual reproduction by transverse fission. Little information is available on their population genetic diversity and structure. Here, the clonal structure of populations of the fissiparous sea cucumber Stichopus chloronotus has been investigated using nine microsatellite loci and a random sampling, at different spatial (intra-reef and inter-reef) and temporal (inter-season and inter-year) scales. Our findings highlight the importance of asexual reproduction in maintaining these populations, and the prevalence of the "initial seedling recruitment" strategy (ISR), leading to a high stability of clonal composition over seasons and years. It also seemed that clonal propagation was limited to the reef scale (<10 km) while reefs were connected by sexual dispersal. This is the first time that clonal structure in sea cucumbers has been studied at such a fine scale, with a specific sampling strategy. It provides key findings on the genetic diversity and structure of fissiparous sea cucumbers, which will be useful for the management of wild populations and aquaculture.


Comparison of synthetic oviductal fluid and G1/G2 medium under low-1 oxygen atmosphere on embryo production and pregnancy rates in Nelore (Bos indicus) cattle.
In this work, we evaluated whether embryo development and pregnancy rates would be affected by culturing bovine Bos indicus embryos in Synthetic Oviductal Fluid with amino acids (SOFaa) or G1/G2 sequential medium under a low-oxygen atmosphere. Using Ovum Pick Up, we obtained 1,538 oocytes, divided into G1/G2 (n = 783) and SOFaa (n = 755). No difference was observed for blastocyst development among the groups (27.8% ± 14.6 and 34.9% ± 20.0 for G1/G2 and SOFaa respectively, p > 0.05). Transferring the embryos (n = 450) from both groups to recipients resulted in similar pregnancy rates for the G1/G2 (38.4% n = 78/203) compared to the SOFaa (39.7% n = 98/247). Our findings confirm that Bos indicus embryos cultured in SOFaa and G1/G2 under low-oxygen atmosphere have similar in vitro (blastocyst rate) and in vivo (pregnancy rate) developmental capacity. However, embryos cultured in G1/G2 medium have higher cleavage than those cultured in SOFaa medium.


Autoradiographic distribution of the D1 agonist [3H]SKF 38393, in the rat brain and spinal cord. Comparison with the distribution of D2 dopamine receptors.
The regional distribution of the specific D1 agonist [3H]SKF 38393 (SKF 38393, 2,3,4,5-tetra-hydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine) has been studied autoradiographically in the rat CNS. The binding of [3H]SKF 38393 to striatal sections was saturable, stereospecific, reversible, of high affinity (Kd = 9.9 nM) and partly sodium sensitive; it occurred at a single population of sites and possessed the pharmacological characteristics of the dopamine D1 receptor. The highest levels of [3H]SKF 38393 binding sites were found in the caudate-putamen, nucleus accumbens, olfactory tubercle and substantia nigra. Moderately high concentrations of the [3H]ligand were observed in the amygdala, endopyriform nucleus, nucleus olfactorius anterior, lateral septum, primary olfactory cortex, cerebellum (molecular layer) and spinal cord. An intermediate labelling was found in the thalamus, habenula, subthalamic nucleus, hypothalamus, ventral tegmental area, superior colliculus, hippocampus and cerebral cortex. Moderate levels of [3H]SKF 38393 binding were observed in the globus pallidus and arcuate nucleus. The autoradiographic distribution of [3H]SKF 38393 overlapped with that of [3H]N,n-propylnorapomorphine, a radioligand which labels the D2 dopamine receptors, in a number of dopamine-rich brain areas but there were several areas which exhibited a high density of [3H]SKF 38393 binding sites but undetectable concentrations of [3H]N,n-propylnorapomorphine. Moreover, in the spinal cord, the subregional localization of these [3H]ligands clearly differed. Intrastriatal injection of ibotenic acid caused a large decrease in [3H]SKF 38393 and [3H]N,n-propylnorapomorphine binding in the striatum and provoked a reduction of [3H]SKF 38393 but not [3H]N,n-propylnorapomorphine binding in the substantia nigra confirming the view that nigral D1 but not D2 receptors are located on striatonigral fibres.


Use of computerized clinical support systems in medical settings: United States, 2001-03.
This report presents information on the use of electronic clinical systems to support patient care in physician offices and hospital emergency and outpatient settings. Percentages of hospital emergency and outpatient departments with electronic patient medical records and automated drug dispensing systems are presented by selected hospital characteristics for 2001-02. Percentages of physicians using electronic patient billing records, electronic patient medical records, and computerized prescription order entry systems are presented by selected physician characteristics for 2003. Data are from provider induction interviews from the National Ambulatory Medical Care Survey (NAMCS) and the National Hospital Ambulatory Medical Care Survey (NHAMCS), the ambulatory care component of the National Health Care Survey (NHCS). The NHCS is a family of provider-based surveys that collect information on the care provided in various medical care settings. During 2001-03, electronic medical records were used less often in physician offices (17 percent) than in hospital emergency (31 percent) and outpatient departments (29 percent). In physician offices, information technology was more frequently used for billing patients (73 percent) than for maintaining medical records electronically (17 percent) or ordering prescriptions electronically (8 percent). Automated drug dispensing systems were available in hospital emergency departments (40 percent) more frequently than in outpatient departments (18 percent).


Short-term soil bioassays may not reveal the full toxicity potential for nanomaterials; bioavailability and toxicity of silver ions (AgNO₃) and silver nanoparticles to earthworm Eisenia fetida in long-term aged soils.
This study investigated if standard risk assessment hazard tests are long enough to adequately provide the worst case exposure for nanomaterials. This study therefore determined the comparative effects of the aging on the bioavailability and toxicity to earthworms of soils dosed with silver ions and silver nanoparticles (Ag NP) for 1, 9, 30 & 52 weeks, and related this to the total Ag in the soil, Ag in soil pore water and earthworm tissue Ag concentrations. For ionic Ag, a classical pattern of reduced bioavailability and toxicity with time aged in the soil was observed. For the Ag NP, toxicity increased with time apparently driven by Ag ion dissolution from the added Ag NPs. Internal Ag in the earthworms did not always explain toxicity and suggested the presence of an internalised, low-toxicity Ag fraction (as intact or transformed NPs) after shorter aging times. Our results indicate that short-term exposures, without long-term soil aging, are not able to properly assess the environmental risk of Ag NPs and that ultimately, with aging time, Ag ion and Ag NP effect will merge to a common value.


Prevalence of Functional Gastrointestinal Disorders in School Children and Adolescents.
The epidemiology of functional gastrointestinal disorders (FGIDs) in developed and developing countries involves a high prevalence of constipation and irritable bowel syndrome. This study examined the prevalence of functional gastrointestinal disorders in schoolchildren and adolescents in Colombia using the Rome III criteria. A cross-sectional study was performed on Colombian children between 8 and 17 years old. The Spanish version of the Questionnaire on Pediatric Gastrointestinal Symptoms-Rome III Version self-report form was answered by students from two schools. The prevalence of FGID was calculated and correlation tests were conducted among the variables analyzed. A total of 864 children with a mean age of 12.5±2.5 years were analyzed; 50.7% were female. Two hundred and fifty-nine children (30%) had at least one FGID, and of these, 163 were female (62.9%). Sixty-nine children had two or more FGIDs (8%). Functional constipation was the most prevalent disorder (13.2%), followed in order by abdominal migraine (8.3%), irritable bowel syndrome (6.9%), and aerophagia (3.1%). A significantly higher prevalence of FGID was observed in females (p=0.000). No significant difference was observed between the age groups or type of school they attended. The overall prevalence of FGID in the sample was 30%, with functional constipation being the most common. These results are similar to those of other prevalence studies reported elsewhere.


Meeting report: national workshops for the communication of air pollution and health information: summary of four workshops in different regions of Europe.
AIRNET was a thematic network project (2002-2004) initiated to stimulate the interaction between researchers in air pollution and health in Europe. As part of AIRNET's communication strategy, a standardized workshop model was developed to organize national meetings on air pollution and health (AIRNET network days) . Emphasis was given to tailor the national workshop information and related activities to the specific needs of a wider range of stakeholders (e.g., policy makers, nongovernmental organizations, industry representatives) . In this report we present an overview of the results of four workshops held in western, northern, central/eastern, and southern regions of Europe in 2004. Overall, workshop experiences indicated that by actively involving participants in the planning of each meeting, AIRNET helped create an event that addressed participants' needs and interests. A wide range of communication formats used to discuss air pollution and health also helped stimulate active interaction among participants. Overall, the national workshops held by AIRNET offered a way to improve communication among the different stakeholders. Because a broad stakeholder involvement in decision making can positively affect the development of widely supported policies, such meetings should be continued for Europe and elsewhere.


IVF recovery of mutant mouse lines using sperm cryopreserved with mtg in cryovials.
Modification of cryoprotective medium (CPM) R18S3 (18% raffinose and 3% skim milk) by addition of monothioglycerol (MTG) or L-glutamine (Glu) has been shown to improve in vitro fertilization (IVF) using mouse sperm cryopreserved in cryostraws. However, whether these CPMs can be applied effectively to sperm cryopreserved in cryovials is unknown. The study was to determine the comparative effectiveness of using R18S3, R18S3+Glu (100mM and 87 mM), or R18S3+MTG (477 µM) to cryopreserve various sample volumes of mouse sperm in cryovials and cryostraws. This study compared the effects of different CPMs on motility of fresh and frozen-thawed C57BL/6J sperm and on IVF rate of C57BL/6J sperm cryopreserved in different CPMs and containers with different volumes, and then used technologies developed to cryopreserve and recover sperm of knockout mouse lines on inbred C57BL/6 backgrounds. Glutamine at 100 mM inhibited, but MTG at 477 µM protected, fresh sperm motility significantly (P < 0.05). Sperm cryopreserved in R18S3+MTG had significantly better (P < 0.05) post-thaw progressive motility and IVF rate than when cryopreserved in R18S3 alone, R18S3+Glu (100 mM), or RSGlu87 (15.7% raffinose, 2.6% skim milk, and 87 mM L-glutamine). There was no significant difference in IVF rates among sperm cryopreserved with R18S3+MTG in cryovials or in cryostraws (P > 0.05). Sperm from 63 knockout mouse lines on C57BL/6 backgrounds cryopreserved using R18S3+MTG in cryovials were all recovered successfully to genotypically-confirmed offspring. Mouse sperm on C56BL/6 backgrounds can be successfully cryopreserved in cryovials using R18S3+MTG.


Identification and characterization of KCASH2 and KCASH3, 2 novel Cullin3 adaptors suppressing histone deacetylase and Hedgehog activity in medulloblastoma.
Medulloblastoma is the most common pediatric malignant brain tumor, arising from aberrant cerebellar precursors' development, a process mainly controlled by Hedgehog (Hh) signaling pathway. Histone deacetylase HDAC1 has been recently shown to modulate Hh signaling, deacetylating its effectors Gli1/2 and enhancing their transcriptional activity. Therefore, HDAC may represent a potential therapeutic target for Hh-dependent tumors, but still little information is available on the physiological mechanisms of HDAC regulation. The putative tumor suppressor REN(KCTD11) acts through ubiquitination-dependent degradation of HDAC1, thereby affecting Hh activity and medulloblastoma growth. We identify and characterize here two REN(KCTD11) homologues, defining a new family of proteins named KCASH, as "KCTD containing, Cullin3 adaptor, suppressor of Hedgehog." Indeed, the novel genes (KCASH2(KCTD21) and KCASH3(KCTD6)) share with REN(KCTD11) a number of features, such as a BTB domain required for the formation of a Cullin3 ubiquitin ligase complex and HDAC1 ubiquitination and degradation capability, suppressing the acetylation-dependent Hh/Gli signaling. Expression of KCASH2 and -3 is observed in cerebellum, whereas epigenetic silencing and allelic deletion are observed in human medulloblastoma. Rescuing KCASHs expression reduces the Hedgehog-dependent medulloblastoma growth, suggesting that loss of members of this novel family of native HDAC inhibitors is crucial in sustaining Hh pathway-mediated tumorigenesis. Accordingly, they might represent a promising class of endogenous "agents" through which this pathway may be targeted.


[ATPase and regulatory properties of skeletal muscle myosin in Citellus undulatus susliks during various stages of winter hibernation].
The paper is devoted to the study on enzymatic and regulatory properties of skeletal muscle myosin of hibernating ground squirrels (Citellus undulatus) for the purpose of elucidating the contribution of myosin and myosin filaments to the change of physiological state of the animal at the different stages of hibernation (hibernation, arousing, winter activity). It has been revealed that actin-activated ATPase activity of myosins of hibernating and arousing ground squirrels is less by 60 and 20%, correspondingly, than that of the myosin of active animals. In the absence of troponin-tropomyosin complex the Ca(2+)-sensitivity of actomyosins of the hibernating and arousing ground squirrels is less by 60 and 55%, correspondingly, than that of the actomyosin of active animals. The results obtained by us point to the essential decrease of functional properties of the main contractile protein, myosin upon hibernation, which evidently contributes to the inhibition of moving capacity of skeletal muscles at this period.


Comparing data on the reproduction of Varroa destructor.
Varroa destructor reproductive success is considered an important character for determining the resistance of honey bees to this mite parasite. However, most of the published data are not comparable due to the different methods of ascertaining and reporting reproduction. A recently published technique that involves reconstructing mite families in older worker brood gives repeatable and reliable parameters. This methodology was used to compare various categories of reproduction of approximately 1,000 V. destructor females in each of three studies on Africanized bees in Brazil and Mexico and European bees in England. The most objective and useful measure was the determination of the number of viable females per female that had invaded the worker brood in singly infested cells, which was denominated the "effective reproduction rate". Viable females are those that can reach the adult stage and have a mate available. The effective reproduction rate in worker brood was 0.64, 0.73 and 1.01 in Brazil, Mexico and England, respectively. Standardization of reproduction determination techniques would make published data comparable and much more useful.


Physiologically interpretable prediction equations for spirometric indexes.
The need for ethnic-specific reference values of lung function variables (LFs) is acknowledged. Their estimation requires expensive and laborious examinations, and therefore additional use of results in physiology and epidemiology would be profitable. To this end, we proposed a form of prediction equations with physiologically interpretable coefficients: a baseline, the onset age (A0) and rate (S) of LF decline, and a height coefficient. The form was tested with data from healthy, nonsmoking Poles aged 18-85 yr (1,120 men, 1,625 women) who performed spirometry maneuvers according to American Thoracic Society criteria. The values of all the coefficients (also A0) for several LFs were determined with regression of LF on patient's age and deviation of patient's height from the mean height in the year group of this patient. S values for forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), peak expiratory flow, and maximal expiratory flow at 75% of FVC (MEF75) were very similar in both sexes (1.03+/-0.07%/yr). FEV1/FVC declines four to five times slower. S for MEF25 appeared age dependent. A0 was smallest (28-32 yr) for MEF25 and FEV1. About 50% of each age subgroup (18-40, 41-60, 61-85 yr) exhibited LFs below the mean, and 4-6% were below the 5th percentile lower limits of normal, and thus the form of equations proposed in the paper appeared appropriate for spirometry. Additionally, if this form is accepted, epidemiological and physiological comparison of different LFs and populations will be possible by means of direct comparison of the equation coefficients.


When cellular networks run out of control: global dysregulation of the RNAi machinery in human pathology and therapy.
RNA interference (RNAi) is an evolutionarily conserved fundamental cellular mechanism of potent gene and genome regulation whose misfunction is associated with numerous major human pathologies, from metabolic disorders and viral infections to cancers. Over the past 5 years, compelling evidence has been accumulated that this association is provided by dysregulations of specific mi(cro)RNAs and the ensuing aberrant expression of their target genes. Moreover, a string of interesting reports has now added proof that human disorders are also frequently characterized by global alterations in the RNAi machinery, comprising irregular expression and function of the key protein players Drosha, DGCR8, Exportin-5, Dicer, TRBP, and Argonaute. Here, we comprehensively review these emerging findings in the specific contexts of cancers and infections with viral pathogens and, in addition, describe related observations in preclinical gene/RNAi therapy studies. Finally, we also thoroughly discuss the relevance of these results for future basic RNAi research as well as for the looming clinical translation of RNAi-based technologies and therapeutic concepts.