Allicin, a natural antimicrobial defence substance from garlic, inhibits DNA gyrase activity in bacteria.
Allicin (diallylthiosulfinate) is a potent antimicrobial substance, produced by garlic tissues upon wounding as a defence against pathogens and pests. Allicin is a reactive sulfur species (RSS) that oxidizes accessible cysteines in glutathione and proteins. We used a differential isotopic labelling method (OxICAT) to identify allicin targets in the bacterial proteome. We compared the proteomes of allicin-susceptible Pseudomonas fluorescens Pf0-1 and allicin-tolerant PfAR-1 after a sublethal allicin exposure. Before exposure to allicin, proteins were in a predominantly reduced state, with approximately 77% of proteins showing less than 20% cysteine oxidation. Protein oxidation increased after exposure to allicin, and only 50% of proteins from allicin-susceptible Pf0-1, but 65% from allicin-tolerant PfAR-1, remained less than 20% oxidised. DNA gyrase was identified as an allicin target. Cys433 in DNA gyrase subunit A (GyrA) was approximately 6% oxidized in untreated bacteria. After allicin treatment the degree of Cys433 oxidation increased to 55% in susceptible Pf0-1 but only to 10% in tolerant PfAR-1. Allicin inhibited E. coli DNA gyrase activity in vitro in the same concentration range as nalidixic acid. Purified PfAR-1 DNA gyrase was inhibited to greater extent by allicin in vitro than the Pf0-1 enzyme. Substituting PfAR-1 GyrA into Pf0-1 rendered the exchange mutants more susceptible to allicin than the Pf0-1 wild type. Taken together, these results suggest that GyrA was protected from oxidation in vivo in the allicin-tolerant PfAR-1 background, rather than the PfAR-1 GyrA subunit being intrinsically less susceptible to oxidation by allicin than the Pf0-1 GyrA subunit. DNA gyrase is a target for medicinally important antibiotics; thus, allicin and its analogues may have potential to be developed as gyrase inhibitors, either alone or in conjunction with other therapeutics.


Circ-ILF2 in oral squamous cell carcinoma promotes cisplatin resistance and induces M2 polarization of macrophages.
Cisplatin (CDDP) chemoresistance is one of the predominant factors in oral squamous cell carcinoma (OSCC) treatment failure. Uncovering the mechanisms underlying CDDP resistance is of great importance in OSCC therapy. Circular RNAs (circRNAs) are a newly discovered class of noncoding RNAs, which are reported to participate in the progression of various diseases, including cancer. However, the function of circRNAs in CDDP resistance in OSCC remains unclear. Quantitative reverse transcription PCR was used to search for different circRNAs between OSCC cell lines and CDDP-resistant cell lines. The results showed that circ-ILF2 expression was higher in CDDP-resistant OSCC cell lines. The stability of circ-ILF2 was also confirmed using RNase R and actinomycin D assays. Functional experiments, including cytotoxicity, apoptosis and growth rate assays, showed that upregulation of circ-ILF2 contributes to CDDP resistance. Luciferase reporter-gene, RNA pull-down and quantitative real-time PCR (RT-qPCR) assays showed that circ-ILF2 functions as a microRNA sponge for miR-1252. Luciferase reporter assays, RNA pull-down, RT-qPCR and Western blotting showed that miR-1252 directly targeted and regulated the expression of KLF8. Circ-ILF2 plays an important role in CDDP resistance in OSCC. Circ-ILF2 exerts its function through the miR-1252/KLF8 pathway. In addition, tumour-associated macrophages (TAM) play important roles in cancer progressions, our results showed that circ-ILF2 in OSCC cells induced the M2 polarization of macrophages which provided new thoughts on immunotherapy. Our results suggest that circ-ILF2 may represent a potential therapeutic target in CDDP-resistant OSCC.


Ultraviolet photoproducts of tryptophan can act as dioxin agonists.
Tryptophan (TRP) has been implicated in the mechanism of induction of microsomal aryl hydrocarbon hydroxylase (AHH) activity by UV light. UV irradiation of TRP resulted in formation of chloroform-soluble photoproducts that can specifically bind to the Ah receptor (AhR), an intracellular protein that mediates the induction of AHH activity by xenobiotics. Gel retardation analysis revealed that the TRP photoproducts, like other AHH inducers, complex to its specific DNA recognition site. Transient transfection experiments also demonstrated that the photoproducts could induce gene expression in an AhR-dependent manner. Thus, induction of AHH activity by UV light could be attributed, at least in part, to the formation of TRP photoproducts, which appear to act as AhR ligands.


Population bottlenecks and nonequilibrium models in population genetics. III. Genic homozygosity in populations which experience periodic bottlenecks.
The amount of variability in a population that experiences repeated restrictions in population size has been calculated. The restrictions in size occur cyclically with a fixed cycle length. Analytical formulas for describing the gene identity at any specific time in the expanded and restricted phases of the cycle, and for the average and second moment of the gene identity, have been derived. It is shown that the level of genetic diversity depends critically on the two parameters that account for the population size, mutation rate and the time of duration for each of the two phases in the cycle. If one or both of these composite parameters are small, the gene diversity will be much reduced, and population gene diversity will then be predictable from knowledge of the harmonic mean population size over the entire cycle. If these parameters take on intermediate values, diversity changes constantly during the cycle, fluctuating steadily from a high to a low value and back again. If these parameters are large, gene diversity will fluctuate rapidly between extreme values and will stay at the extremes for long periods of time.


Retinal stimulates ATP hydrolysis by purified and reconstituted ABCR, the photoreceptor-specific ATP-binding cassette transporter responsible for Stargardt disease.
Many substrates for P-glycoprotein, an ABC transporter that mediates multidrug resistance in mammalian cells, have been shown to stimulate its ATPase activity in vitro. In the present study, we used this property as a criterion to search for natural and artificial substrates and/or allosteric regulators of ABCR, the rod photoreceptor-specific ABC transporter responsible for Stargardt disease, an early onset macular degeneration. ABCR was immunoaffinity purified to apparent homogeneity from bovine rod outer segments and reconstituted into liposomes. All-trans-retinal, a candidate ligand, stimulates the ATPase activity of ABCR 3-4-fold, with a half-maximal effect at 10-15 microM. 11-cis- and 13-cis-retinal show similar activity. All-trans-retinal stimulates the ATPase activity of ABCR with Michaelis-Menten behavior indicative of simple noncooperative binding that is associated with a rate-limiting enzyme-substrate intermediate in the pathway of ATP hydrolysis. Among 37 structurally diverse non-retinoid compounds, including nine previously characterized substrates or sensitizers of P-glycoprotein, only four show significant ATPase stimulation when tested at 20 microM. The dose-response curves of these four compounds are indicative of multiple binding sites and/or modes of interaction with ABCR. Two of these compounds, amiodarone and digitonin, can act synergistically with all-trans-retinal, implying that they interact with a site or sites on ABCR different from the one with which all-trans-retinal interacts. Unlike retinal, amiodarone appears to interact with both free and ATP-bound ABCR. Together with clinical observations on Stargardt disease and the localization of ABCR to rod outer segment disc membranes, these data suggest that retinoids, and most likely retinal, are the natural substrates for transport by ABCR in rod outer segments. These observations have significant implications for understanding the visual cycle and the pathogenesis of Stargardt disease and for the identification of compounds that could modify the natural history of Stargardt disease or other retinopathies associated with impaired ABCR function.


Who is using snus? - Time trends, socioeconomic and geographic characteristics of snus users in the ageing Swedish population.
The prevalence of smoking in Sweden has decreased in recent decades, and is now among the lowest in the world. During the same period, the use of Swedish moist oral snuff, a smokeless tobacco called snus, has increased. Few studies have evaluated time trends of the socioeconomic and geographic characteristics of snus users in Sweden. This paper contributes to filling that gap. This study utilized the Linnaeus Database, which links national registers with comprehensive individual data on socioeconomic status (SES) to health data from a large ongoing health survey, the Västerbotten Intervention Programme (VIP). The VIP targets the entire middle-aged population of Västerbotten county at ages 40, 50 and 60 years with yearly cross-sectional surveys including self-reported data on tobacco habits. Time trends of snus use among 92,563 VIP-participants across different areas of residence and smoking groups were investigated graphically. Logistic regression was performed to estimate the associations between SES and geographical variables and current use versus non-use of snus. Overall, in parallel to decreasing smoking, the increasing trend of snus use in this middle-aged population continues, particularly in 40-year-olds. In both genders, the highest prevalence of snus use was observed among previous smokers. The prevalence of snus use also increased over time among smokers, and was consistently higher compared to those who had never smoked. Among males - both those who had never smoked and previous smokers - low education (OR 1.21, 95%CI 1.06-1.40 and OR 1.28, 95%CI 1.14-1.43), living alone (OR 1.16, 95%CI 1.07-1.27 and OR 1.13, 95%ci 1.04-1.23), low income and living in rural areas was associated with using snus, while this was not seen among male current smokers. Among women, living alone was associated with using snus irrespective of smoking habits. Among female smokers, the OR for snus use increased with higher education. A disadvantaged social profile and also higher prevalence in rural areas is observed among male snus users who had never smoked or were previous smokers. Among male smokers there was no association between SES and use of snus. The prevalence of snus use among women is increasing, but is still considerably lower than that of men. The association between snus and SES characteristics is less pronounced among women, although snus is clearly linked to living alone. These patterns should be taken into consideration in tobacco control policies.


Rare cardiac defect in Holt-Oram syndrome.
Holt-Oram syndrome is an autosomal dominant condition associated with skeletal malformations of the upper limbs, and congenital heart disease. Approximately 40% of cases represent new mutations. Defective development of the embryonic radial ray (e.g. aplasia, hypoplasia, fusion, other anomalous development) results in a wide spectrum of phenotypes, including triphalangeal or absent thumbs, foreshortened arms and phocomelia. The syndrome is associated with defective development of cardiac structures that results in atrial septal defect (ASD), most commonly the secundum type, heart block of varying degrees, or both. We report a rare cardiac defect patent ductus arteriosus (PDA) and ventricular septal defect (VSD) in a case of Holt-Oram syndrome.


Skin texture aging trend analysis using dermoscopy images.
To date, the degree of skin damage caused by diverse factors, such as aging and persistent sunlight exposure, has been evaluated based on the personal experience and knowledge of dermatologists because there is no standard method for objective evaluation. If a standard method were available, patients could obtain more consistent information about their skin condition, and hence perform more effective treatment of the skin damage. In this paper, we demonstrate how to establish a standard method using dermoscopy images of subjects of various ages. We focus on three body parts, specifically the face, neck, and hands, and extract various skin texture features to quantitatively and objectively represent the skin condition. We construct a model for skin damage evaluation based on various skin texture features. To accomplish this objective, we consider various features from face, neck, and hand dermoscopy images, including texture length, width and depth, cell area, the number of cells in a fixed region, radius ratio of inscribed and circumscribed circles of a wrinkle cell, and average perimeter of a wrinkle cell. In this study, a wrinkle cell represents the smallest skin region enclosed by textures. We then perform a linear regression for texture features based on subject age. A dermoscopy image can be automatically analyzed by extracting skin texture features. We demonstrate aging trends by performing linear regression on these features. Based on this result, a quantitative and objective evaluation of the skin condition can be provided. We proposed several new skin texture features and developed algorithms to accurately extract them. We analyzed these features and demonstrated their age-related change trends by using graphs and charts. We believe that our result can be used as a standard method for evaluating degrees of skin damage. Moreover, we believe that our proposed method can be applied in various areas, such as performance evaluation of certain skin products.


Hormone replacement therapy and cardioprotection: what is good and what is bad for the cardiovascular system?
The incidence of cardiovascular diseases (CVDs) increases after menopause and at any age postmenopausal women have a significantly higher incidence of CVD compared to premenopausal women. Several epidemiological findings suggest the causative pathogenetic role of ovarian hormone deficiency in the development of CVD in women. Ovarian hormones have several potential protective effects on the cardiovascular system and despite several observational studies have shown the beneficial effect of estrogens and estrogen/progestin associations on CVD, at the present, after the findings of randomized studies, the effect of hormone replacement therapy (HRT) in the prevention of CVD is still under debate. The randomized studies (Heart and Estrogen/Progestin Replacement Study [HERS] and Women's Health Initiative [WHI]) found largely concordant results with the observational studies except for the divergent findings about coronary heart disease (CHD). The discrepancy between the two arms of the WHI study suggests that two factors, time to initiation of HRT since menopause and estrogen/progestin associations, are of pivotal importance to explain the widely divergent findings on the cardiovascular effects of observational studies and randomized clinical studies. Basic science and animal studies together with clinical investigations and the results of clinical studies are concordant in suggesting that a long time since menopause is associated with a reduced protective effect of estrogens while the unfavorable effects upon coagulation remain unaltered. In early postmenopausal women, like the ones included in the observational studies, ovarian hormone replacement may be cardioprotective because of the responsiveness of the endothelium to estrogens that also buffer the detrimental effects upon coagulation. In late postmenopausal women ovarian hormones have either a null effect or even a detrimental effect because of the predominance of the procoagulant or plaque-destabilizing effects over the vasoprotective effects. Therefore, HRT has beneficial cardiovascular effects in younger women while it may have detrimental effect on coagulative balance and vascular inflammation and has little effect on cardiovascular functions in older women.


Electron Transfer and Modification of Oligosilanylsilatranes and Related Derivatives.
Several silatranyl -substituted oligosilanes were prepared starting from bis(trimethylsilyl)silatranylsilanide. Electrochemical and theoretical investigations of some oligosilanes revealed that electrooxidation occurs by formation of a short-lived cation radical. This species undergoes structural relaxation to form a pair of conformers, with endo and exo relationships with respect to the Si-N interaction. Reaction of a 1,4-disilatranyl-1,4-disilanide with 1,2-dichlorotetramethyldisilane gave a mixture of cis and trans diastereomers of a cyclohexasilane with the trans isomer showing a diminished Si-N distance.


Prokaryotic adenylate cyclase toxin stimulates anterior pituitary cells in culture.
Bordetella pertussis synthesizes a variety of virulence factors including a calmodulin-dependent adenylate cyclase (AC) toxin. Treatment of anterior pituitary cells with this AC toxin resulted in an increase in cellular cAMP levels that was associated with accelerated exocytosis of growth hormone (GH), prolactin, adrenocorticotropic hormone (ACTH), and luteinizing hormone (LH). The kinetics of release of these hormones, however, were markedly different; GH and prolactin were rapidly released, while LH and ACTH secretion was more gradually elevated. Neither dopamine agonists nor somatostatin changed the ability of AC toxin to generate cAMP (up to 2 h). Low concentrations of AC toxin amplified the secretory response to hypophysiotrophic hormones. We conclude that bacterial AC toxin can rapidly elevate cAMP levels in anterior pituitary cells and that it is this response that explains the subsequent acceleration of hormone release.


Sperm competitive ability evolves in response to experimental alteration of operational sex ratio.
In naturally polygamous organisms such as Drosophila, sperm competitive ability is one of the most important components of male fitness and is expected to evolve in response to varying degrees of male-male competition. Several studies have documented the existence of ample genetic variation in sperm competitive ability of males. However, many experimental evolution studies have found sperm competitive ability to be unresponsive to selection. Even direct selection for increased sperm competitive ability has failed to yield any measurable changes. Here we report the evolution of sperm competitive ability (sperm defense-P1, offense-P2) in a set of replicate populations of Drosophila melanogaster subjected to altered levels of male-male competition (generated by varying the operational sex ratio) for 55-60 generations. Males from populations with female-biased operational sex ratio evolved reduced P1 and P2, without any measurable change in the male reproductive behavior. Males in the male-biased regime evolved increased P1, but there was no significant change in P2. Increase in P1 was associated with an increase in copulation duration, possibly indicating greater ejaculate investment by these males. This study is one of the few to provide empirical evidence for the evolution of sperm competitive ability of males under different levels of male-male competition.


miR‑210 regulates esophageal cancer cell proliferation by inducing G2/M phase cell cycle arrest through targeting PLK1.
micro (mi)RNAs are short regulatory RNAs that negatively modulate protein expression at the post‑transcriptional level, and are being considered as novel therapeutic targets for the treatment of cancer. In the present study, an elevated expression level of circulating miR‑210 was observed in patients with esophageal squamous cell carcinoma (ESCC) for the first time, to the best of our knowledge, and the induction of miR‑210 under hypoxic conditions in ESCC was confirmed. Cell counting kit‑8 assay and bromodeoxyuridine incorporation assay indicated that miR‑210 markedly inhibited the proliferation of ESCC cells. In addition, the effect of miR‑210 on the cell cycle was examined. Transfection of miR‑210 resulted in a significant increase in the proportion of cells in G2/M phase. Polo‑like kinase 1 (PLK1) was investigated as a candidate target of miR‑210, which is a critical regulator of cell cycle transmission at multiple levels. It was demonstrated that miR‑210 reduced the levels of PLK1 protein by binding the 3' untranslated region of its mRNA. The results of the present study demonstrated that miR‑210 inhibited the proliferation of ESCC cells by inducing G2/M phase cell cycle arrest, and these effects of miR‑210 were mediated by the targeting of PLK1.


Rac2 GTPase deficiency depletes BCR-ABL+ leukemic stem cells and progenitors in vivo.
Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease (MPD) initiated by p210-BCR-ABL-mediated transformation of hematopoietic stem cells (HSCs). Inhibition of the ABL kinase alone is not sufficient to eradicate leukemic stem cells (LSCs). We have previously shown that the deficiency of Rac2 GTPase signaling, but not Rac1, in p210-BCR-ABL-transduced hematopoietic cells prolonged survival of mice with MPD. Here we demonstrate that absence of Rac2 GTPase prolongs survival of HSC-initiated, inducible Scl/p210-BCR-ABL (Scl/p210) binary transgenic mice, it induces apoptosis, and, unlike in normal HSC and progenitor (HSC/P), impairs LSC and progenitor (LSC/P) proliferation in vivo. As a result, Rac2 deficiency causes functional exhaustion of the LSC pool in vivo. This defect is not due to impaired interaction with the hematopoietic microenvironment as reflected by its unaltered adhesion, migration, and homing to recipient organs. In summary, Rac2 deficiency exhausts the LSC pool in vivo through impairment of oncogene-induced proliferation and survival signals.


The effects of glyphosate exposure on gene transcription and immune function of the silkworm, Bombyx mori.
Glyphosate is a widely used herbicide and crop desiccant. However, whether its extensive use has any effect on the species diversity of nontarget organisms is still unclear. In this study, we used the silkworm, Bombyx mori, as the research subject, and performed RNA sequencing to analyze the transcriptional profile of silkworm midgut after exposure to glyphosate at 2975.20 mg/L (a concentration commonly used at mulberry fields). A total of 125 significantly differentially expressed genes (DEGs) were detected in the midgut of glyphosate-exposed silkworm (q < 0.05), of which 53 were upregulated and 72 were downregulated. Gene ontology enrichment analysis showed that the DEGs were mainly enriched in biological process, cellular component, and molecular function. Kyoto encyclopedia of genes and genomes analysis showed that the differential genes were mainly related to oxidative stress, nutrient metabolism, and immune defense pathways, including oxidative stress-related Cat and Jafrac1, nutrient metabolism-related Fatp and Scpx, and immune-related CYP6AN2, UGT40B4, CTL11, serpin-2, and so forth. Experimental verification showed that glyphosate exposure led to a 4.35-fold increase in the mortality of silkworm after Beauveria bassiana infection, which might be caused by the decreased PO (phenoloxidase) activity and impaired immunity. These results provide evidence for the potential effects of residue glyphosate on the physiological functions of silkworm, and also provide a reference for the biosafety evaluation of glyphosate.


Conformational preferences of modified nucleobases in RNA aptamers and their effect on Förster resonant energy transfer.
Förster resonant energy transfer (FRET) can be utilized in the study of tertiary structures of RNA aptamers, which bind specific fluorophoric ligands to form a fluorogenic aptamer complex. By introducing the emissive nucleobase analog 4-cyanoindole into the fluorogenic Chili RNA aptamer a FRET pair was established. The interpretation of studies aiming to investigate those tertiary structures using FRET, however, relies on prior knowledge about conformational properties of the nucleobase, which govern exciton transfer capabilities. Herein we employed classical molecular dynamics combined with Förster exciton theory to elucidate the preferred orientation relative to proximate bases and the influence on exciton transfer efficiency in multiple substitution sites. We did this by comparing the chromophoric distances emergent from MD simulations with experimental FRET data based on structural data of the native aptamer. We present the outlined methodology as a means to reliably evaluate future nucleobase analogue candidates in terms of their structural behavior and emergent exciton transfer properties as exemplified in the study of the preferred orientation of 4-cyanoindole in the Chili RNA aptamer.


Establishing ionising radiation safety culture during interventional cardiovascular procedures.
The safety culture of an interventional laboratory was investigated in terms of the application of the three cardinal principles of radiation protection, namely distance, time and shielding. The application of these principles was observed and recorded with a radiation safety-culture checklist that was compiled by consulting international recommendations. The checklist evaluated the optimal compliance, especially with reference to monitoring of staff exposure, distance from the X-ray source, fluoroscopy techniques pertaining to frame rate, protective devices and personal shielding. The effective radiation dose was measured to the eyes, thyroid, hands and feet of the cardiologist, nurse, floor nurse and radiographer by means of finger dosimeters that were readily available from the local radiation-protection dosimetry service. The results, after observing 11 procedures, indicated the absence of table and ceiling-suspended shields, and the distance of the cardiologist's and scrub nurse's feet from the X-ray tube were between 16 and 68 cm, with a mean distance of 59.7 and 58.5 cm, respectively. Most staff (91%) wore the dosimeter inside the lead apron at the collar level without eye protection. The highest dosimeter values recorded were 0.73 mSv to the hand of the cardiologist, 0.45 mSv to the eye of the cardiologist, 0.65 mSv to the hand of the scrub nurse, 0.54 mSv to the eye of the scrub nurse and 0.52 mSv to the foot of the scrub nurse. The dosimeter value to the radiographer's thyroid was 0.42 mSv. The dosimeter readings confirmed the highest doses were to the scrub nurse and hand of the interventionalist. The safety culture was non-compliant in terms of staff distance being too close to the X-ray tube, the absence of ceiling and table screens, the theatre door not always being completely closed, and for staff without lead eye glasses, wearing dosimeters outside the lead apron at the collar level.


Ca2+/calmodulin-dependent protein kinase IIβ and IIδ mediate TGFβ-induced transduction of fibronectin and collagen in human pulmonary fibroblasts.
It is now clear that in addition to activating several complex kinase pathways (Smad, MAP kinase, PI3 kinase), TGFβ also acts by elevating cytosolic Ca2+ concentration within human pulmonary fibroblasts. Ca2+/calmodulin-dependent protein kinase II (CamK II) is also known to regulate gene expression in fibroblasts. In this study, we examined the interactions between calcium signaling, activation of CamK and other kinases, and extracellular matrix (ECM) gene expression. Human pulmonary fibroblasts were cultured and stimulated with artificially generated Ca2+ pulses in the absence of TGFβ, or with TGFβ (1 nM) or vehicle in the presence of various blockers of Ca2+ signaling. PCR and Western blotting were used to measure gene expression and protein levels, respectively. We found that Ca2+ pulses in the absence of TGFβ increased ECM gene expression in a pulse frequency-dependent manner, and that blocking Ca2+ signaling and the CamK II pathway significantly reduced TGFβ-mediated ECM gene expression, without having any effects on other kinase pathways (Smad, PI3 kinase, or MAP kinase). We also found that TGFβ elevated the expression of CamK IIβ and CamK IIδ, while siRNA silencing of those two subtypes significantly reduced TGFβ-mediated expression of collagen A1 and fibronectin 1. Our data suggest that TGFβ induces the expression of CamK IIβ and CamK IIδ, which in turn are activated by TGFβ-evoked Ca2+ waves in a frequency-dependent manner, leading to increased expression of ECM proteins.


Executive dysfunction in remitted late-life depression: Juntendo University Mood Disorder Projects (JUMP).
The authors aimed to investigate whether remitted adult and elderly major depressive disorder patients show different patterns of executive dysfunction. Executive functions of 20 euthymic major depressive disorder patients and 29 healthy comparison subjects were evaluated using the Behavioral Assessment of the Dysexecutive Syndrome. Relative to adult patients and healthy comparison subjects, euthymic elderly patients were more impaired in the subtest of Modified Six Elements. Since the regions most implicated in this subtest are the medial prefrontal, the anterior cingulate, and the dorsolateral prefrontal areas, the authors conclude that dysfunctions of such frontal neural networks remain unresolved even in the remission phase of late-life depression.


Bioluminescent imaging of HPV-positive oral tumor growth and its response to image-guided radiotherapy.
The treatment paradigms for head and neck squamous cell cancer (HNSCC) are changing due to the emergence of human papillomavirus (HPV)-associated tumors possessing distinct molecular profiles and responses to therapy. Although patients with HNSCCs are often treated with radiotherapy, preclinical models are limited by the ability to deliver precise radiation to orthotopic tumors and to monitor treatment responses accordingly. To better model this clinical scenario, we developed a novel autochthonous HPV-positive oral tumor model to track responses to small molecules and image-guided radiation. We used a tamoxifen-regulated Cre recombinase system to conditionally express the HPV oncogenes E6 and E7 as well as a luciferase reporter (iHPV-Luc) in the epithelial cells of transgenic mice. In the presence of activated Cre recombinase, luciferase activity, and by proxy, HPV oncogenes were induced to 11-fold higher levels. In triple transgenic mice containing the iHPV-Luc, K14-CreER(tam), and LSL-Kras transgenes, tamoxifen treatment resulted in oral tumor development with increased bioluminescent activity within 6 days that reached a maximum of 74.8-fold higher bioluminescence compared with uninduced mice. Oral tumors expressed p16 and MCM7, two biomarkers associated with HPV-positive tumors. After treatment with rapamycin or image-guided radiotherapy, tumors regressed and possessed decreased bioluminescence. Thus, this novel system enables us to rapidly visualize HPV-positive tumor growth to model existing and new interventions using clinically relevant drugs and radiotherapy techniques.


Therapy-refractory gastrointestinal motility disorder in a child with c-kit mutations.
Constipation and fecal impaction are frequent and distressing complaints in pediatric gastroenterology. Especially in neurologically handicapped children, treatment of severe forms of slow-transit constipation (STC) can be difficult. In the majority of cases, STC is of unknown etiology. However, in recent years, there is growing evidence that interstitial cells of Cajal (ICCs), which serve as electrical pacemakers and generate spontaneous electrical slow waves in the gastrointestinal tract, might play an important role in the pathophysiology of STC. It remains unclear whether morphological ICC alterations seen in affected patients are based on congenital developmental anomalies, or whether they are a consequence of long-term constipation with secondary damage of the gastrointestinal nervous system. To the best of our knowledge, we present the first case of a patient with histological alterations in ICC morphology who displayed multiple alterations of c-kit at the level of mRNA. The protein encoded by c-kit is the receptor tyrosine kinase Kit (CD117), which is crucial for development and function of ICCs. Therefore, these findings provide a new explanation for congenital alterations of ICC development that result in gastrointestinal motility disorders.


Regulation of Chlamydia spreading from the small intestine to the large intestine via an immunological barrier.
The obligate intracellular bacterium Chlamydia is a genital tract pathogen that can also colonize the gastrointestinal tract for long periods. The long-lasting colonization is dependent on chlamydial spreading from the small intestine to the large intestine. We previously reported that a mutant Chlamydia was able to activate an intestinal barrier for blocking its own spreading to the large intestine. In the current study, we used the mutant Chlamydia colonization model to confirm the intestinal barrier function and further to determine the immunological basis of the barrier with gene-deficient mice. Recombination activating gene 1-/- mice failed to block the mutant Chlamydia spreading, while mice deficient in toll-like receptors, myeloid differentiation primary response 88 or stimulator of interferon genes still blocked the spreading, suggesting that the intestinal barrier function is dependent on lymphocytes that express antigen receptors. Mice deficient in CD4, but not CD8 nor μ chain failed to prevent the chlamydial spreading, indicating a protective role of CD4+ cells in the intestinal barrier. Consistently, adoptive transfer of CD4+ T cells reconstituted the intestinal barrier in CD4-/- mice. More importantly, CD4+ but not CD8+ T cells nor B cells restored the intestinal barrier function in recombination activating gene 1-/- mice. Thus, CD4+ T cells are necessary and sufficient for maintaining the intestinal barrier function, indicating that the spread of an intracellular bacterium from the small intestine to the large intestine is regulated by an immunological barrier. This study has also laid a foundation for further illuminating the mechanisms by which a CD4+ T cell-dependent intestinal barrier regulates bacterial spreading in the gut.


Nurses' self-care strategies: A mapping review.
Nurses have long received recommendations to employ self-care behaviors to ensure their ability to remain in the profession; however, nurses are rarely asked what self-care behaviors are beneficial. This literature review aimed to map studies on nurses' self-care strategies to provide an understanding of how these strategies are addressed in the literature and to identify gaps in need of additional exploration. Searches were conducted in accordance with published mapping review methodologies across MEDLINE, Embase, CINAHL, Scopus, PsycINFO, Web of Science, and identified influential nursing journals. Included studies were from the United States, peer-reviewed, and described self-care strategies related to nurses or the nursing profession. Thirty-five articles were identified for inclusion and were evaluated based on agreed-upon criteria to define how the concept of self-care was studied and applied to nurses. Three main categories were identified in this review: self-care and stress; self-care and burnout; and self-care and leadership, workplace factors, and physical activity. This mapping review revealed an abundance of literature related to self-care recommendations for nurses; however, there remains a dearth of studies investigating how nurses utilize self-care behaviors independently to improve their own professional well-being. Recommendations for next steps in research in this area are included.


Functional effects of the antigen glatiramer acetate are complex and tightly associated with its composition.
Glatiramer acetate (Copaxone®; GA) is a non-biological complex drug for multiple sclerosis. GA modulated thousands of genes in genome-wide expression studies conducted in THP-1 cells and mouse splenocytes. Comparing GA with differently-manufactured glatiramoid Polimunol (Synthon) in mice yielded hundreds of differentially expressed probesets, including biologically-relevant genes (e.g. Il18, adj p<9e-6) and pathways. In human monocytes, 700+ probesets differed between Polimunol and GA, enriching for 130+ pathways including response to lipopolysaccharide (adj. p<0.006). Key differences were confirmed by qRT-PCR (splenocytes) or proteomics (THP-1). These studies demonstrate the complexity of GA's mechanisms of action, and may help inform therapeutic equivalence assessment.


Deletions and recombinations in the core region of hepatitis B virus genotype E strains from asymptomatic blood donors in Guinea, west Africa.
The prevalence of hepatitis B virus (HBV) surface antigen (HBsAg) chronic carriage in west Africa is the highest in the world, but its molecular epidemiology remains relatively poorly investigated. Plasma samples from random asymptomatic carriers of HBsAg in Conakry, Guinea, were studied and the complete genome sequences of 81 strains were obtained. Three additional samples from Kumasi, Ghana, were also included in the analysis. Phylogenetic analyses confirmed the dominance of genotype E (95.1%), including 8.6% of strains (viral load, 5x10(3)-2.6x10(8) IU ml(-1)) comprising dominant variants with large deletions in the core region and minority wild-type variants. The presence of two different patterns of deletions in two and four donors suggested targeted genome fragility between nt 1979 and 2314. The remaining sequences included one subgenotype A3 (1%) and six A/E recombinant forms (4-7%). A/E strains with identical points of recombination in three donors suggested strongly that these recombinant HBV strains are circulating and transmitted in the population. Recombination points were concentrated in the core gene. The detection of similar A/E recombinant strains in Ghana suggested a geographical extension of recombinant HBV to the region. The quasispecies of one additional Ghanaian strain sequenced in the pre-surface/surface region resolved into dominant clones of either the A or E genotype, but also three different patterns of A/E recombinant variants. The observation that both deletions of genotype E strains and A/E recombination points are mostly located in the core gene at specific positions indicates a region of the genome where genetic rearrangements preferentially take place.


Evidence for pituitary regulation of placental lactogen in the rat.
The present studies examined whether the pituitary hormones, particularly prolactin, have any role in the regulation of rat placental lactogen (rPL) secretion. Total rPL was measured using a lymphoma cell bioassay. Hypophysectomy on day 13 of pregnancy increased total serum rPL levels above those of intact control rats and delayed for 3 days the decline in rPL usually seen by day 14. To test the effect of early hypophysectomy on rPL secretion, a delayed implantation model was used. Hypophysectomy was carried out on day 3, progesterone (2 mg) was given daily until day 8 and oestrone (1.0 micrograms) was given on day 8. This initiated implantation on day 9, which is 4 days later than normal. Rats hypophysectomized on day 3 had high serum levels of rPL (10-13 mg/l) 7 days after initiation of implantation compared with control values of 2-3 mg/l, and these levels remained raised for the duration of the experiment. Termination of maintenance injections of steroids did not affect the high levels of rPL for several days, even though there was fetal and placental resorption. When a pair of anterior pituitaries was transplanted under the kidney capsule on day 13 (day 9 of development) of rats hypophysectomized on day 3, serum rPL levels still increased for the next 3 days. However, unlike similarly treated rats not receiving transplants, rPL levels fell rapidly thereafter and were only 5% of those in control animals 1 week later.(ABSTRACT TRUNCATED AT 250 WORDS)


Evoked response audiometry in scrub typhus: prospective, randomised, case-control study.
To investigate the hypothesis of cochlear and retrocochlear damage in scrub typhus, using evoked response audiometry. Prospective, randomised, case-control study. The study included 25 patients with scrub typhus and 25 controls with other febrile illnesses not known to cause hearing loss. Controls were age- and sex-matched. All subjects underwent pure tone audiometry and evoked response audiometry before commencing treatment. Six patients presented with hearing loss, although a total of 23 patients had evidence of symmetrical high frequency loss on pure tone audiometry. Evoked response audiometry found significant prolongation of absolute latencies of wave I, III, V, and wave I-III interpeak latency. Two cases with normal hearing had increased interpeak latencies. These findings constitute level 3b evidence. Findings were suggestive of retrocochlear pathology in two cases with normal hearing. In other patients, high frequency hearing loss may have led to altered evoked response results. Although scrub typhus appears to cause middle ear cochlear and retrocochlear damage, the presence of such damage could not be fully confirmed by evoked response audiometry.


Stretchable Loudspeaker using Liquid Metal Microchannel.
Considering the various applications of wearable and bio-implantable devices, it is desirable to realize stretchable acoustic devices for body-attached applications such as sensing biological signals, hearing aids, and notification of information via sound. In this study, we demonstrate the facile fabrication of a Stretchable Acoustic Device (SAD) using liquid metal coil of Galinstan where the SAD is operated by the electromagnetic interaction between the liquid metal coil and a Neodymium (Nd) magnet. To fabricate a liquid metal coil, Galinstan was injected into a micro-patterned elastomer channel. This fabricated SAD was operated simultaneously as a loudspeaker and a microphone. Measurements of the frequency response confirmed that the SAD was mechanically stable under both 50% uniaxial and 30% biaxial strains. Furthermore, 2000 repetitive applications of a 50% uniaxial strain did not induce any noticeable degradation of the sound pressure. Both voice and the beeping sound of an alarm clock were successfully recorded and played back through our SAD while it was attached to the wrist under repeated deformation. These results demonstrate the high potential of the fabricated SAD using Galinstan voice coil in various research fields including stretchable, wearable, and bio-implantable acoustic devices.


Evidence for articular cartilage regeneration in MRL/MpJ mice.
A major clinical problem in Orthopaedics is the repair of traumatic articular cartilage lesions. The MRL/MpJ strain of mice has the remarkable ability to regenerate ear hole punch wounds seamlessly including the scarless replacement of multiple tissues. The objective of this study was to assess whether articular cartilage defects repair or regenerate in the MRL/MpJ 'healer' strain of mice. Full thickness and partial thickness lesions were introduced into trochlear groove articular cartilage of MRL/MpJ and C57Bl/6 mice, a control strain that does not undergo ear hole regeneration. The wound sites were assessed 6 weeks and 12 weeks post-surgery using a histological scoring scheme and immunohistochemistry for markers of articular cartilage including proteoglycan, collagen II and collagen VI. The partial thickness lesions did not repair in either strain. However, at both 6 weeks and 12 weeks timepoints the MRL/MpJ mice had a superior healing response of full thickness lesions with abundant chondrocytes and an extracellular matrix rich in proteoglycan, collagen II and collagen VI at the wound site. At the 12 week timepoint the enhanced cartilage healing was restricted to male MRL/MpJ mice. In contrast, the C57Bl/6 control strain produced an extracellular matrix at the wound site that, overall, had significantly less matrix proteoglycan and collagen II. Male MRL/MpJ mice appear to possess an intrinsic ability to 'regenerate' articular cartilage. Understanding the biochemical and genetic basis for articular cartilage regeneration may open up new treatment options for traumatic articular cartilage defects.


The integration of dominance and social bonding in primates.
Social dominance is usually viewed from the perspective of intragroup competition over access to limited resources. The present paper, while not denying the importance of such competition, discusses the dominance concept among monkeys and apes in the context of affiliative bonding, social tolerance, and the reconciliation of aggressive conflicts. Two basic proximate mechanisms are supposed to provide a link between dominance and interindividual affiliation, namely, formalization of the dominance relationship (i.e., unequivocal communication of status), and conditional reassurance (i.e., the linkage of friendly coexistence to formalization of the relationship). Ritualized submission is imposed upon losers of dominance struggles by winners; losers are offered a "choice" between continued hostility or a tolerant relationship with a clearly signalled difference in status. If these two social mechanisms are lacking, aggression is bound to have dispersive effects. In their presence, aggression becomes a well-integrated, even constructive component of social life. In some higher primates this process of integration has reached the stage where status differences are strongly attenuated. In these species, sharing and trading can take the place of overt competition. The views underlying this "reconciled hierarchy" model are only partly new, as is evident from a review of the ethological literature. Many points are illustrated with data on a large semi-captive colony of chimpanzees (Pan troglodytes), particularly data related to striving for status, reconciliation behavior, and general association patterns. These observations demonstrate that relationships among adult male chimpanzees cannot be described in terms of a dichotomy between affiliative and antagonistic tendencies. Male bonding in this species has not been achieved by an elimination of aggression, but by a set of powerful buffering mechanisms that mitigate its effects. Although female chimpanzees do exhibit a potential for bonding under noncompetitive conditions, they appear to lack the buffering mechanisms of the males.


Mitochondrial DNA damage and dysfunction, and oxidative stress are associated with endoplasmic reticulum stress, protein degradation and apoptosis in high fat diet-induced insulin resistance mice.
Recent studies showed a link between a high fat diet (HFD)-induced obesity and lipid accumulation in non-adipose tissues, such as skeletal muscle and liver, and insulin resistance (IR). Although the mechanisms responsible for IR in those tissues are different, oxidative stress and mitochondrial dysfunction have been implicated in the disease process. We tested the hypothesis that HFD induced mitochondrial DNA (mtDNA) damage and that this damage is associated with mitochondrial dysfunction, oxidative stress, and induction of markers of endoplasmic reticulum (ER) stress, protein degradation and apoptosis in skeletal muscle and liver in a mouse model of obesity-induced IR. C57BL/6J male mice were fed either a HFD (60% fat) or normal chow (NC) (10% fat) for 16 weeks. We found that HFD-induced IR correlated with increased mtDNA damage, mitochondrial dysfunction and markers of oxidative stress in skeletal muscle and liver. Also, a HFD causes a change in the expression level of DNA repair enzymes in both nuclei and mitochondria in skeletal muscle and liver. Furthermore, a HFD leads to activation of ER stress, protein degradation and apoptosis in skeletal muscle and liver, and significantly reduced the content of two major proteins involved in insulin signaling, Akt and IRS-1 in skeletal muscle, and Akt in liver. Basal p-Akt level was not significantly influenced by HFD feeding in skeletal muscle and liver. This study provides new evidence that HFD-induced mtDNA damage correlates with mitochondrial dysfunction and increased oxidative stress in skeletal muscle and liver, which is associated with the induction of markers of ER stress, protein degradation and apoptosis.


Studies on the efficacy of hyperbaric rendering procedures in inactivating bovine spongiform encephalopathy (BSE) and scrapie agents.
The efficacy of the procedures in use at the two rendering plants in the Netherlands was assessed on a laboratory-scale using procedures that simulated the pressure cooking part of the rendering process. A pool of bovine spongiform encephalopathy (BSE)-infected brainstem from the United Kingdom and a pool of scrapie-infected brainstem from Dutch sheep were used to spike the rendering materials. The mixtures were subjected to various time-temperature combinations of hyperbaric heat treatment related to the conditions used in Dutch rendering plants in the early 1990s, and to the combination of 20 minutes at 133 degrees C required by the EU Directive on rendering of 1996. The efficacy of the procedures in inactivating BSE or scrapie infectivity was measured by titrating the materials before and after heat treatment in inbred mice, by combined intracerebral and intraperitoneal inoculations at limiting dilutions. Two independent series of experiments were carried out. The design of the study allowed for minimum inactivations of up to 2.2 log (2.0 in the second series) to be measured in the diluted infective material and 3.1 log in the undiluted material. After 20 minutes at 133 degrees C there was a reduction of BSE infectivity of about 2.2 log in the first series (with some residual infectivity detected), and in the second series more than 2.0 log (with no residual infectivity detected). With undiluted brain material there was an inactivation of about 3.0 log (with some residual infectivity detected). With the same procedure, scrapie infectivity was reduced by more than 1.7 log in the first series and by more than 2.2 log in the second series. With undiluted brain material there was an inactivation of more than 3.1 log. In each case no residual scrapie infectivity was detected. The BSE agent consistently appeared to be more resistant to heat inactivation procedures than the scrapie agent, particularly at lower temperatures and shorter times.


Characterization of one alpha 2-macroglobulin with anticomplementary activity from pregnant women.
In 15 pregnant women during the third term of pregnancy, the immunomodulatory property of alpha 2-macroglobulin (alpha 2M) was initially detected by measuring the inhibitory effect on immune complement-dependent haemolysis of serum alpha 2M fractions obtained by gel filtration. By a two-step chromatography procedure consisting of gel filtration followed by anion-exchange chromatography, different sub-forms of alpha 2M in serum were separated. Amongst them, it was shown that the inhibition of complement activity was almost exclusively linked to one particular subform. Additional studies revealed that the observed effect was not due to proteases bound to alpha 2M during clotting since, by using protease-specific inhibitors, no change was observed in complement inhibition. This subform, though present at very low levels in control sera, appeared in strikingly increased amounts during the third trimester of pregnancy (35 mg/l) and comprised between 3 and 5% of the total alpha 2M. Results show that the increase of alpha 2M anticomplementary activity is linked to the increase in alpha 2M levels in serum.


The mitogen-activated protein kinase phosphatase PHS1 regulates flowering in Arabidopsis thaliana.
Arabidopsis PHS1, initially known as an actor of cytoskeleton organization, is a positive regulator of flowering in the photoperiodic and autonomous pathways by modulating both CO and FLC mRNA levels. Protein phosphorylation and dephosphorylation is a major type of post-translational modification, controlling many biological processes. In Arabidopsis thaliana, five genes encoding MAPK phosphatases (MKP)-like proteins have been identified. Among them, PROPYZAMIDE HYPERSENSITIVE 1 (PHS1) encoding a dual-specificity protein tyrosine phosphatase (DsPTP) has been shown to be involved in microtubule organization, germination and ABA-regulated stomatal opening. Here, we demonstrate that PHS1 also regulates flowering under long-day and short-day conditions. Using physiological, genetic and molecular approaches, we have shown that the late flowering phenotype of the knock-out phs1-5 mutant is linked to a higher expression of FLOWERING LOCUS C (FLC). In contrast, a decline of both CONSTANS (CO) and FLOWERING LOCUS T (FT) expression is observed in the knock-out phs1-5 mutant, especially at the end of the light period under long-day conditions when the induction of flowering occurs. We show that this partial loss of sensitivity to photoperiodic induction is independent of FLC. Our results thus indicate that PHS1 plays a dual role in flowering, in the photoperiodic and autonomous pathways, by modulating both CO and FLC mRNA levels. Our work reveals a novel actor in the complex network of the flowering regulation.


Association of a Novel Mutation in the Plasmodium falciparum Chloroquine Resistance Transporter With Decreased Piperaquine Sensitivity.
Amplified copy number in the plasmepsin II/III genes within Plasmodium falciparum has been associated with decreased sensitivity to piperaquine. To examine this association and test whether additional loci might also contribute, we performed a genome-wide association study of ex vivo P. falciparum susceptibility to piperaquine. Plasmodium falciparum DNA from 183 samples collected primarily from Cambodia was genotyped at 33716 genome-wide single nucleotide polymorphisms (SNPs). Linear mixed models and random forests were used to estimate associations between parasite genotypes and piperaquine susceptibility. Candidate polymorphisms were evaluated for their association with dihydroartemisinin-piperaquine treatment outcomes in an independent dataset. Single nucleotide polymorphisms on multiple chromosomes were associated with piperaquine 90% inhibitory concentrations (IC90) in a genome-wide analysis. Fine-mapping of genomic regions implicated in genome-wide analyses identified multiple SNPs in linkage disequilibrium with each other that were significantly associated with piperaquine IC90, including a novel mutation within the gene encoding the P. falciparum chloroquine resistance transporter, PfCRT. This mutation (F145I) was associated with dihydroartemisinin-piperaquine treatment failure after adjusting for the presence of amplified plasmepsin II/III, which was also associated with decreased piperaquine sensitivity. Our data suggest that, in addition to plasmepsin II/III copy number, other loci, including pfcrt, may also be involved in piperaquine resistance.


Comparison of the efficacy and safety of once-daily dosing and on-demand use of udenafil for type 2 diabetic patients with erectile dysfunction.
We compared the efficacy and safety between once-daily dosing and on-demand use of udenafil for type 2 diabetic patients with erectile dysfunction (ED). A multi-center, randomized, open-label, parallel-group, 12-week study was conducted. 161 patients who improved with on-demand 200 mg of udenafil according to Sexual Encounter Profile (SEP) diary Question 2 and 3 (Q2 and Q3) were randomized into 200 mg on-demand (n = 80) or 50 mg once-daily (n = 81) dosing groups for 8 weeks. The dosing period was followed by a 4-week treatment-free period. The primary efficacy endpoint was the change of the International Index of Erectile Function (IIEF) erectile function domain (EFD) score. The secondary efficacy endpoints included changes to the SEP diary Q2, Q3, IIEF Q3, Q4, other domains of IIEF, Global Assessment Question, and shift to the normal rate (EFD ≥ 26). Vascular endothelial markers were also assessed. The IIEF-EFD score of both groups improved after 8 weeks of treatment (P < 0.0001). There was no statistically significant difference between two groups. Improvement was not maintained after the treatment-free follow-up period. Similar results were observed in the secondary efficacy endpoints. There was also no significant difference in vascular endothelial markers. Daily udenafil was well-tolerated, and there was no significant difference in the adverse drug reactions and adverse events between the two groups. Flushing and headache were the most frequent adverse events. Both regimens improved ED in diabetic patients and were well-tolerated. Further studies are needed to assess the effect of daily udenafil treatment in diabetic patients.


Development and initial validation of the Influences on Patient Safety Behaviours Questionnaire.
Understanding the factors that make it more or less likely that healthcare practitioners (HCPs) will perform certain patient safety behaviors is important in developing effective intervention strategies. A questionnaire to identify determinants of HCP patient safety behaviors does not currently exist. This study reports the development and initial validation of the Influences on Patient Safety Behaviors Questionnaire (IPSBQ) based on the Theoretical Domains Framework. Two hundred and thirty-three HCPs from three acute National Health Service Hospital Trusts in the United Kingdom completed the 34-item measure focusing on one specific patient safety behavior (using pH as the first line method for checking the position of a nasogastric tube). Confirmatory factor analysis (CFA) was undertaken to generate the model of best fit. The final questionnaire consisted of 11 factors and 23 items, and CFA produced a reasonable fit: χ² (175) = 345.7, p < 0.001; CMIN/DF = 1.98; GFI = 0.90 and RMSEA = 0.06, as well as adequate levels of discriminant validity, and internal consistency (r = 0.21 to 0.64). A reliable and valid theoretically underpinned measure of determinants of HCP patient safety behavior has been developed. The criterion validity of the measure is still unknown and further work is necessary to confirm the reliability and validity of this measure for other patient safety behaviors.


Current biochemical understanding regarding the metabolism of acinetobactin, the major siderophore of the human pathogen Acinetobacter baumannii, and outlook for discovery of novel anti-infectious agents based thereon.
Covering: 1994 to 2019Owing to the rapid increase in nosocomial infections by antibiotic-resistant Acinetobacter baumannii and the paucity of effective treatment options for such infections, interest in the virulence factors involved in its successful dissemination and propagation in the human host have escalated in recent years. Acinetobacin, a siderophore of A. baumannii, is responsible for iron acquisition under nutritional depravation and has been shown to be one of the key virulence factors for this bacterium. In this Highlight, recent findings regarding various chemical and biological aspects of acinetobactin metabolism closely related to the fitness of A. baumannii at the infection sites have been described. In addition, several notable efforts for identifying novel anti-infectious agents based thereon have been discussed.


Effective Partnerships in School Reform: Lessons Learned From the Midwest Child-Parent Center Expansion.
The Midwest Expansion of the Child-Parent Center Education Program (MCPC) is a pre-K to 3rd grade intervention program aimed at improving economically disadvantaged children's school success by enhancing continuity in instruction and increasing parental involvement. Opened in Chicago in the 1960s, this school reform model has undergone significant changes and is currently being expanded in five demographically heterogeneous school districts in Illinois and Minnesota. This paper describes the collaborative process that has contributed to effective implementation of the expansion project within at-risk communities. Three themes of collaboration are emphasized: (a) improving the quality of data, (b) establishing and maintaining implementation fidelity, and (c) using research to inform practice. We discuss lessons learned from our partnerships with a number of collaborators including those involved with implementing the pre-K program, researching and evaluating the project, and providing professional development to teachers. Consideration is given to advancing the field of implementation science by successfully initiating and enhancing exemplary partnerships in comprehensive school reforms through effective research design.


Staphylococcus aureus clumping factor B mediates biofilm formation in the absence of calcium.
Staphylococcus aureus is the leading cause of nosocomial infections and a major cause of community-acquired infections. Biofilm formation is a key virulence determinant in certain types of S. aureus infection, especially those involving inserted medical devices. We found in a previous study that the calcium chelators sodium citrate and EGTA inhibit biofilm formation in certain strains of S. aureus but actually augment biofilm formation in other strains. Even two closely related strains, Newman and 10833, exhibited strikingly different biofilm phenotypes in the presence of calcium chelators, in that biofilm formation was inhibited in Newman but augmented in 10833. We also found that the surface protein clumping factor B (ClfB) plays a role in this phenomenon. In this study, we confirm that ClfB is required for biofilm formation under calcium-depleted conditions. We investigated the post-translational regulation of ClfB-mediated biofilm formation and found evidence that both calcium and the protease aureolysin disrupt established ClfB-dependent biofilms. Finally, we investigated the genetic basis for the biofilm-negative phenotype in strain Newman versus the biofilm-positive phenotype in strain 10833 under calcium-depleted conditions and found that strain 10833 contains a deletion that results in a stop codon within the aureolysin gene (aur). When 10833 expressed Newman aur, surface-associated ClfB and the ability to form a biofilm in chelating conditions was lost. Thus, the positive effect of chelating agents on biofilm formation in certain strains can be explained by increased ClfB activity in the absence of calcium and the discrepancy in the response of strains 10833 and Newman can be explained by point mutations in aur. This study reveals a previously unknown, to our knowledge, role for ClfB in biofilm formation and underscores the potential for striking phenotypic variability resulting from minor differences in strain background.


Urinary N-nitrosoproline excretion: a further evaluation of the nitrosamine hypothesis of gastric carcinogenesis in precancerous conditions.
Measurement of N-nitroso compounds in gastric juice by different methods has given conflicting results. In order to resolve this controversy, we have assessed endogenous nitrosation by the independent N-nitrosoproline excretion test in subjects who had previously undergone gastric juice analysis by one of these methods. Ten Polya gastrectomy, 10 pernicious anaemia and nine matched control subjects were fed 380 mg of nitrate in beetroot juice and 500 mg proline. N-nitrosoproline (N-Pro) synthesised intragastrically from these precursors, and quantitatively excreted by the kidneys, was measured in 24 hour urine samples (collection checked by creatinine clearance). N-Pro excretion (mean +/- SEM) was reduced (p less than 0.01) in pernicious anaemia (1.1 +/- 0.8 ng/day) compared with matched control (18.0 +/- 7.2 ng/day), and also tended to be lower (NS) in polya gastrectomy (3.2 +/- 2.3 ng/day). Twenty four hour intragastric pH was monitored on a separate occasion in 23 of the 29 subjects; 13 were hypoacidic (pH greater than 4 greater than 50% of 24 hours) and 10 were acidic. N-Pro yields were reduced (p less than 0.01) in the hypoacidic group (0.9 +/- 0.6 ng/day) compared with the acidic group (17.9 +/- 6.6 ng/day), and N-Pro was negatively associated with mean intragastric pH (tau = -0.53, p = 0.001). We conclude that endogenous synthesis of this specific N-nitroso compound is favoured by low rather than high pH. These results are concordant with those previously reported in gastric juice from the same subjects and suggest that nitrosation is chemically rather than bacterially mediated, contrary to the nitrosamine hypothesis of gastric carcinogenesis.


Head size, weaponry, and cervical adaptation: Testing craniocervical evolutionary hypotheses in Ceratopsia.
The anterior cervical vertebrae form the skeletal connection between the cranial and postcranial skeletons in higher tetrapods. As a result, the morphology of the atlas-axis complex is likely to be shaped by selection pressures acting on either the head or neck. The neoceratopsian (Reptilia:Dinosauria) syncervical represents one of the most highly modified atlas-axis regions in vertebrates, being formed by the complete coalescence of the three most anterior cervical vertebrae. In ceratopsids, the syncervical has been hypothesized to be an adaptation to support a massive skull, or to act as a buttress during intraspecific head-to-head combat. Here, we test these functional/adaptive hypotheses within a phylogenetic framework and critically examine the previously proposed methods for quantifying relative head size in the fossil record for the first time. Results indicate that neither the evolution of cranial weaponry nor large head size correlates with the origin of cervical fusion in ceratopsians, and we, therefore, reject both adaptive hypotheses for the origin of the syncervical. Anterior cervical fusion has evolved independently in a number of amniote clades, and further research on extant groups with this peculiar anatomy is needed to understand the evolutionary basis for cervical fusion in Neoceratopsia.


Adaptable DNA interactions regulate surface triggered self assembly.
DNA-mediated multivalent interactions between colloidal particles have been extensively applied for their ability to program bulk phase behaviour and dynamic processes. Exploiting the competition between different types of DNA-DNA bonds, here we experimentally demonstrate the selective triggering of colloidal self-assembly in the presence of a functionalised surface, which induces changes in particle-particle interactions. Besides its relevance to the manufacturing of layered materials with controlled thickness, the intrinsic signal-amplification features of the proposed interaction scheme make it valuable for biosensing applications.


Cerebral Emboli Generation During Off-Pump Coronary Artery Bypass Grafting With a Clampless Device versus Partial Clamping of the Ascending Aorta.
: It is not known whether use of a clampless facilitating device during proximal graft anastamosis decreases intraoperative cerebral emboli in patients with mild atherosclerotic ascending aorta (AAA) having off-pump coronary artery bypass. : After intraoperative epiaortic ultrasound showed no more than mild (grade I-II) atherosclerotic ascending aorta, 20 patients were randomized to receive either partial clamping (PC, n = 10) or the HEARTSTRING clampless device (HS, n = 9) for proximal graft construction on the ascending aorta. Continuous transcranial Doppler monitoring, with capability to discern gaseous from solid particulates, was used intraoperatively to monitor high-intensity transient signals (HITS) in the middle cerebral arteries. Postoperative diffusion-weighted brain magnetic resonance imaging documented old and new ischemic brain lesions. : There were no significant differences between the groups in the number of proximals (P = 0.14), distals (P = 0.4), or intraoperative cell saver transfusions (P = 0.69). The total number of HITS was not significantly different between the PC and HS groups (P = 0.2). However, the number of solid HITS was significantly lower in the HS than in the PC group (2.7 ± 2.6 versus 14.0 ± 8.1; P < 0.001). The number of gaseous emboli in the HS group was fourfold greater when a mister-blower rather than a suction device was used to clear blood away from the HS site. Postoperatively, there were no deaths, myocardial infarctions, or clinical strokes observed in either group. Diffusion-weighted cerebral magnetic resonance imaging revealed no statistical difference between groups for new infarct lesions. : Use of the HS device during off-pump coronary artery bypass was associated with significantly fewer intraoperative solid emboli in the middle cerebral artery than PC of the ascending aorta.


Studying for a doctorate: developing a strong voice.
Studying for a doctoral degree can be an exciting, invigorating and ultimately satisfying process but equally it can be a long, arduous, frustrating and lonely journey. In this article I speak of my own experience of the process and offer some thoughts about the 'what, why, when and how' of studying at this level. I consider the excitement of developing your own ideas and project alongside the challenges inherent in doctoral level study, such as coping with uncertainty and rarely having a sense of closure, and I offer some practical challenges to help you decide whether doctoral study is for you.


Is urinary tract infection after shock wave lithotripsy an aggravating factor for renal damage?
To prospectively assess if urinary tract infection (UTI) after extracorporeal shock wave lithotripsy (ESWL) enhances immediate renal damage in patients with renal stones. Levels of lactic dehydrogenase (LDH), creatine phosphokinase (CPK), and beta2 microglobulin were measured in the urine of 171 patients, before and after ESWL for treatment of up to 3 cm renal stones. Standard investigation included urine sample before and after treatment. Patients were divided in two groups in accordance to presence of positive bacterial urine sample after ESWL. Changes in biomarker values pre and post- ESWL were assessed via X2 test for both groups and compared between the two groups for post-treatment values. Despite pre-treatment antibiotic therapy, 21.6% of the patients presented with UTI after ESWL. In the group with clear post-treatment urine samples there was a 5.8, 2.2, and 0.7 fold increase of CPK, LDH, and beta2 microglobulin levels after ESWL, respectively (p<0.001), indicating some degree of immediate renal damage. In patients with UTI, the levels of the enzymes elevated by 80, 15, and 2.2 fold after ESWL, respectively, which represents a significantly increased elevation comparing with the sterile urine group (p<0.001). ESWL results in transient immediate renal damage, which is exaggerated in patients with persistent or treatment prompted UTI. Long-term importance of this finding must be determined.


Plasma cholinesterase correlations in acute surgical and critical illness.
The properties of plasma cholinesterase (CHE) are partly undiscovered. Equally unknown are the correlations between changes in CHE and other blood variables during the acute phase response related to acute surgical and critical illness. Data from 432 measurements of CHE and other variables performed in 92 patients were systematically evaluated and processed by regression analysis. There was a strong direct correlation between CHE and albumin (r=0.77, P<0.0001). CHE was also directly correlated to cholesterol, iron binding capacity, hematocrit, prothrombin activity, and inversely correlated to bilirubin and to presence of sepsis or liver dysfunction (P<0.0001 for all). Postoperatively CHE decreased to about 60% of the preoperative value, remaining directly related to it (r=0.69, P<0.0001), and decreasing further in the presence of sepsis or liver dysfunction, with slow reversal of the decrease during recovery from illness. In parenterally fed septic patients the decrease in CHE was moderated by increasing the amino acid dose (P<0.0001). In acute surgical and critical illness CHE mostly behaves as a negative acute phase reactant, independently of the modifications related to other already known factors. This should be taken into account when interpreting the implications of decreased CHE in the clinical setting.


Reduced muscle function in patients with osteoarthritis.
The purpose of this study was to determine whether subjects with knee osteoarthritis (OA) had reduced muscle strength at various muscle lengths, endurance, contraction velocity and functional capacity, compared with control subjects and whether the decrease was related to functional capacity. Forty-five men and 45 women with knee OA were compared with a control group (41 males, 63 females) of similar age for functional capacity, maximal isometric strength (in vivo length-tension relationship) and endurance (in vivo force-time relationship) of knee flexion and extension and maximal angular velocity (in vivo force-velocity relationship) of knee extension. The OA subjects had increased difficulty (2.03 +/- 0.53) and pain (1.65 +/- 0.29) for activities of daily living (ADLs) and significantly lower strength for extension (72%) and flexion (56%), endurance for the quadriceps (203%) and hamstrings (214%) and velocity (128%). The reductions were greater at longer muscle lengths. These data demonstrate that patients with knee OA have reduced muscle function and functional capacity compared to controls.


Self-reported drunkenness among adolescents in four sub-Saharan African countries: associations with adverse childhood experiences.
Consumption of alcohol is associated with acute and chronic adverse health outcomes. There is a paucity of studies that explore the determinants of alcohol use among adolescents in sub-Saharan Africa and, in particular, that examine the effects of adverse childhood experiences on alcohol use. The paper draws on nationally-representative data from 9,819 adolescents aged 12-19 years from Burkina Faso, Ghana, Malawi, and Uganda. Logistic regression models were employed to identify correlates of self-reported past-year drunkenness. Exposure to four adverse childhood experiences comprised the primary independent variables: living in a food-insecure household, living with a problem drinker, having been physically abused, and having been coerced into having sex. We controlled for age, religiosity, current schooling status, the household head's sex, living arrangements, place of residence, marital status, and country of survey. All analyses were conducted separately for males and females. At the bivariate level, all independent variables (except for coerced sex among males) were associated with the outcome variable. Overall, 9% of adolescents reported that they had been drunk in the 12 months preceding the survey. In general, respondents who had experienced an adverse event during childhood were more likely to report drunkenness. In the multivariate analysis, only two adverse childhood events emerged as significant predictors of self-reported past-year drunkenness among males: living in a household with a problem drinker before age 10, and being physically abused before age 10. For females, exposure to family-alcoholism, experience of physical abuse, and coerced sex increased the likelihood of reporting drunkenness in the last 12 months. The association between adverse events and reported drunkenness was more pronounced for females. For both males and females there was a graded relationship between the number of adverse events experienced and the proportion reporting drunkenness. We find an association between experience of adverse childhood events and drunkenness among adolescents in four sub-Saharan African countries. The complex impacts of adverse childhood experiences on young people's development and behavior may have an important bearing on the effectiveness of interventions geared at reducing alcohol dependence among the youth.


Status of low-energy accelerator-based BNCT worldwide and in Argentina.
Existing and active low-energy Accelerator-Based BNCT programs worldwide will be reviewed and compared. In particular, the program in Argentina will be discussed which consists of the development of an Electro-Static-Quadrupole (ESQ) Accelerator-Based treatment facility. The facility is conceived to operate with the deuteron-induced reactions 9Be(d,n)10B and 13C(d,n)14N at 1.45 MeV deuteron energy, as neutron sources. Neutron production target development status is specified. The present status of the construction of the new accelerator development laboratory and future BNCT centre is shown.


Influenza vaccination and the risk of primary cardiac arrest.
Influenza epidemics are associated with an excess of mortality not only from respiratory diseases but also from other causes, and cardiovascular mortality increases abruptly during influenza epidemics, with little evidence of a lag period. In a population-based case-control study, the authors examined whether influenza vaccination was associated with a reduced risk of out-of-hospital primary cardiac arrest (PCA), a major contributor to cardiovascular mortality in the community. Cases of PCA (n = 342) without prior heart disease or life-threatening comorbidity that occurred in King County, Washington, were identified from paramedic incident reports from October 1988 to July 1994. Demographically similar controls (n = 549) were identified from the community by using random digit dialing. Spouses of subjects were interviewed to assess treatment with influenza vaccine during the previous year and other risk factors. After adjustment for demographic, clinical, and behavioral risk factors, influenza vaccination was associated with a reduced risk of PCA (odds ratio = 0.51, 95 percent confidence interval: 0.33, 0.79). The authors suggest that while the association of influenza vaccination with a reduced risk of PCA is consistent with cohort studies of influenza vaccination and total mortality, further studies are needed to determine whether the observed association reflects protection or selection.


Long term trends in oral antidiabetic drug use among children and adolescents in the Netherlands.
The aim of the study was to document long term trends in oral antidiabetic drug (OAD) use among children and adolescents in the Netherlands. A population-based cohort study was conducted using the Dutch PHARMO Database Network. All patients younger than 20 years old with at least one OAD dispensing were identified. Age-adjusted and age-specific incidence (1999-2011) and prevalence (1998-2011) rates of OAD use were calculated. Trends over time were assessed using joinpoint regression software. A subset of PHARMO Database Network (including community pharmacy dispensing records linked to general practitioner data (OPD-GP database)) was used to assess indications for OADs. In 2011, the overall age-adjusted incidence and prevalence rates of OAD use were 20.7/100 000 (95% CI 19.2, 22.1) person-years (PY) and 53.8/100 000 (95% CI 51.5, 56.1) persons, respectively. The average annual percentage change (AAPC) in the overall age-adjusted incidence rates from 1999 to 2011 was 18.9% (95% CI 4.5, 35.2). The incidence and prevalence rates of OAD use were higher among females and older age categories. The increases in rates of OAD use were mainly driven by metformin. For only 50% of the 98 patients in the OPD-GP database, indications for OAD prescriptions were reported with type 1 diabetes (n = 20), type 2 diabetes (n = 16), and overweight/obesity (n = 10). Incidence and prevalence rates of OAD use in children and adolescents substantially increased in the Netherlands, especially among older age categories (10-14 and 15-19 years) and females. The main indications for use of OADs were type 1 and 2 diabetes and overweight/obesity.


Stiffness of the distal tip of bipolar pacing leads.
The stiffness of a bipolar pacing lead, particularly between anode and cathode, may be responsible for myocardial penetration and perforation. Following an unprecedented 7% incidence of high threshold exit block with a single model bipolar ventricular endocardial lead, a study was undertaken to compare pacing lead stiffness between anode and cathode of six models of bipolar leads from two manufacturers; Telectronics (T) and Medtronics (M). Four leads had polyurethane insulation; T 030-284 (Laser Dish), T 329-259 (Cordis, Encor), M 4012 (Target Tip), and M 4004 (Capsure). Two leads had silicone rubber insulation; M 5026 (Capsure) and M 5024 (Capsure SP). All leads were subjected to two stiffness tests. The Tip Deflection Test involved securing the lead at 45 degrees at the indifferent electrode and applying a force to deflect the tip 5 mm. The three point bending test involved placing the lead over two fixed bars in contact with the anode and cathode. Midway a third bar was pushed onto the lead and the force to deflect the lead 2 mm was recorded. The results showed that pacing leads with polyurethane insulation were much stiffer than those with silicone rubber insulation. The T 030-284 because of its construction was found to be the stiffnest. The next stiffnest was the M 4012. Both these leads had an unacceptable incidence of high threshold exit block; 7% with the T 030-284 (89 implants) and 3% with the M 4012 (102 implants). No cases of high threshold exit block were documented with the other four pacing leads and in particular the silicone rubber M 5026 (344 implants).(ABSTRACT TRUNCATED AT 250 WORDS)


Saturated hydrogen saline protects the lung against oxygen toxicity.
Exposure to high oxygen concentrations leads to acute lung injury, including lung tissue and alveolar edema formation, congestion, intra-alveolar hemorrhage, as well as endothelial and epithelial cell apoptosis or necrosis. Several studies have reported that molecular hydrogen is an efficient antioxidant by gaseous rapid diffusion into tissues and cells. Moreover, consumption of water with dissolved molecular hydrogen to a saturated level (hydrogen water) prevents stress-induced cognitive decline in mice and superoxide formation in mice. The purpose of the present study was to investigate the effect of saturated hydrogen saline on pulmonary injury-induced exposure to >98% oxygen at 2.5 ATA for five hours. Adult male Sprague-Dawley (SD) rats were randomly divided into three groups: control group, saline group and saturated hydrogen saline group. Hematoxylin and eosin (H&E) staining were used to examine histological changes. The lung wet to dry (W/D) weight ratio was calculated. The concentration of protein and total cell counts in bronchoalveolar lavage fluid (BALF) were measured. Lactate dehydrogenase (LDH) in serum and BALF were measured by spectrophotometer. The light microscope findings showed that saturated hydrogen saline reduced the impairment when compared with the saline group: Saturated hydrogen saline decreased lung edema, reduced LDH activity in BALF and serum, and decreased total cells and protein concentration in BALF. These results demonstrated that saturated hydrogen saline alleviated hyperoxia-induced pulmonary injury, which was partly responsible for the inhibition of oxidative damage.


Negotiation Practices of Athletic Trainers Employed in the Clinical Setting.
Athletic trainers (ATs) are comparatively underpaid relative to peer health care professionals. Whereas many factors contribute to the salary and benefits of a given employment position, negotiation is a factor of the final salary and benefits package that is achieved. It is unclear to what extent ATs negotiate salary or other terms of employment during the hiring process. To explore the negotiation practices of ATs during the hiring process. Cross-sectional study. Web-based survey. A total of 587 ATs employed in the clinical setting who previously held at least 1 full-time employment position. Independent variables were several demographic factors as well as the current salary range. Dependent variables were participants' responses to various survey items focused on experiences with salary and terms-of-employment negotiation. Summary statistics were used to characterize all variables and multiple χ2 analyses (P < .05) were performed to determine the significant influences of independent variables on negotiation practices. More than half of ATs (57.6%) did not attempt to negotiate their salary, and almost three-quarters of ATs (70.5%) did not negotiate their terms of employment during the hiring process. The most successfully negotiated terms were moving expenses (72.3%) and continuing education funding and reimbursement (62.7%). The influence of demographic factors on negotiation and negotiation success varied, with significant findings for the number of previous full-time employment positions, gender, marital status, salary range, and number of dependents. It is alarming that more than half of ATs did not negotiate salary or terms of employment during the hiring process. Whereas widespread training on negotiation practices is warranted, our findings suggest it would be most beneficial for early-career and female ATs. All ATs must become comfortable with negotiating salary and terms of employment to effect change in the average salary and employment status of those in the profession.


Estimating the Risk of Domestic Water Source Contamination Following Precipitation Events.
Climate change is expected to increase precipitation extremes, threatening water quality. In low resource settings, it is unclear which water sources are most vulnerable to contamination following rainfall events. We evaluated the relationship between rainfall and drinking water quality in southwest Guatemala where heavy rainfall is frequent and access to safe water is limited. We surveyed 59 shallow household wells, measured precipitation, and calculated simple hydrological variables. We compared Escherichia coli concentration at wells where recent rainfall had occurred versus had not occurred, and evaluated variability in the association between rainfall and E. coli concentration under different conditions using interaction models. Rainfall in the past 24 hours was associated with greater E. coli concentrations, with the strongest association between rainfall and fecal contamination at wells where pigs were nearby. Because of the small sample size, these findings should be considered preliminary, but provide a model to evaluate vulnerability to climate change.


[Theater of Life: theory, method and practice].
Theater of Life is an educational model that integrate theater theories and techniques as a strategy for health education. Theater as an educational technique has become a useful tool for the health context. In this work the author discuss the role of social change as an important element in health education and suggests the use of theatrical techniques for it's promotion. Also offers information about the different approaches of popular theatre and popular education incorporated in this model, Theatre of the Oppressed by A. Boal, Popular Education by P. Freire, Poor Theater by J. Grotowoski and Education for Peace by C. Beristain and P. Cascón and explain the basic principles of each one of them. In the methodology section the author explains the different steps for implementing the strategy: solidarity and connection games, story telling technique and script development, presentation and forum. In the practice section the author shares the process of model development and the significance events that had contribute to their elaboration.


Structure of the chloroplast gene for the precursor of the Mr 32,000 photosystem II protein from mustard (Sinapis alba L.).
The nucleotide sequence of the mustard chloroplast gene for the precursor of the Mr 32,000 photosystem II protein is presented. A comparison with the corresponding genes from spinach and Nicotiana debneyi (14) reveals less than 5% nucleotide divergence in the coding region. The derived protein of mustard differs from the corresponding proteins by three amino acid positions at the C-terminus. We have defined the presumed transcription start and termination sites of the mustard gene. Upstream from the start site are sequences typical of a prokaryotic promoter and, also, a sequence that resembles the eukaryotic 'TATA' box. A search for intrastrand base pairing revealed stem-loop secondary structure at the transcription start and termination sites and in the region preceding the presumed promoter. This latter region is a 69-base-pair sequence element unique to the 5'flanking sequence of the mustard gene.


Genetic diversity for root plasticity and nitrogen uptake in wheat seedlings.
Enhancing nitrogen use efficiency (NUE) of wheat is a major focus for wheat breeding programs. NUE may be improved by identifying genotypes that are competitive for nitrogen (N) uptake in early vegetative stages of growth and are able to invest that N in grain. Breeders tend to select high yielding genotypes under conditions of medium to high N supply, but it is not known whether this influences the selection of root plasticity traits or whether, over time, breeders have selected genotypes with higher N uptake efficiency. To address this, genotypes were selected from CIMMYT (1966-1985) and Australian (1999-2007) breeding programs. Genotypes from both programs responded to low N supply by expanding their root surface area through increased total root number and/or length of lateral roots. Australian genotypes were N responsive (accumulated more N under high N than under low N) whereas CIMMYT genotypes were not very N responsive. This could not be explained by differences in N uptake capacity as shown by 15N flux analysis of two representative genotypes with contrasting N accumulation. Expression analysis of nitrate transporter genes revealed that the high-affinity transport system accounted for the majority of root nitrate uptake in wheat seedlings under both low and high N conditions.


[Evaluation of the function of the left ventricle of the heart according to the results of radionuclide ventriculography in the diagnosis of ischemic heart disease].
Radionuclide (99mTc) ventriculography demonstrated a tendency to reduced ejection fraction, mean normalized systolic expulsion rate and myocardial circular fibre shortening rate of coronary patients that was proportionate to the severity of their condition. Myocardial lesion was shown to be heterogeneous and typically involve asynergic areas. As the disease progresses, the disturbance of myocardial contractility grows more severe, and the number of hypokinetic areas increases while that of normo- and hyperkinetic areas declines.


Reprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and sequence-specific RNA biosensors.
In type II CRISPR systems, the guide RNA (gRNA) comprises a CRISPR RNA (crRNA) and a hybridized trans-acting CRISPR RNA (tracrRNA), both being essential in guided DNA targeting functions. Although tracrRNAs are diverse in sequence and structure across type II CRISPR systems, the programmability of crRNA-tracrRNA hybridization for Cas9 is not fully understood. Here, we reveal the programmability of crRNA-tracrRNA hybridization for Streptococcus pyogenes Cas9, and in doing so, redefine the capabilities of Cas9 proteins and the sources of crRNAs, providing new biosensing applications for type II CRISPR systems. By reprogramming the crRNA-tracrRNA hybridized sequence, we show that engineered crRNA-tracrRNA interactions can not only enable the design of orthogonal cellular computing devices but also facilitate the hijacking of endogenous small RNAs/mRNAs as crRNAs. We subsequently describe how these re-engineered gRNA pairings can be implemented as RNA sensors, capable of monitoring the transcriptional activity of various environment-responsive genomic genes, or detecting SARS-CoV-2 RNA in vitro, as an Atypical gRNA-activated Transcription Halting Alarm (AGATHA) biosensor.


Endobronchial valve treatment in chronic obstructive pulmonary disease: A qualitative study of patients' expectations.
Endobronchial valve (EBV) treatment is a treatment option for selected patients with severe chronic obstructive pulmonary disease (COPD) to reduce dyspnea and improve quality of life. However, the procedure is associated with risk of complications, and not all patients achieve the expected outcomes. The present study explores patients' expectations while waiting for EBV treatment. Fifteen patients accepted for EBV treatment at Aarhus University Hospital in Denmark from October 2020 to June 2021 participated in a 30-min, semi-structured interview about (1) the experience of living with COPD and (2) expectations regarding EBV. Four themes were identified: a life of reduced quality; hopes and expectations; information about EBV; and perception of risks. Most patients described their lives as not worth living, and they hoped that EBV would give them their physical and social lives back, while also being aware that EBV would probably not work miracles. Patients' information seeking was influenced by their hopes and expectations to the treatment. They filtered out negative information, focusing on the hope for a positive outcome. This made them willing to run the risk of complications. Living a life of considerably reduced quality, patients might have an increased focus on potential positive effects of EBV treatment while filtering out information about potential side effects. This might bias their decision-making process.


Neural modulation by stimulation.
Spinal cord stimulation (SCS) for the treatment of neuropathic pain is supported by good-quality randomized controlled trials, prospective and retrospective case studies, and observational case series that confirm its efficacy and safety. SCS has been successfully used in various refractory neuropathic pain conditions, including failed back surgery syndrome (FBSS), neuropathic back and leg pain, and complex regional pain syndrome (CRPS) types I and II. According to the Harbour and Miller Scale (2001), the evidence for SCS in FBSS has been classified as grade B, while that for CRPS type I has been classified as grade A. Clinical evidence has shown that compared to conventional pain therapy, more than two-thirds of carefully selected patients treated with SCS achieved sustained pain relief of 50% or more, with minimal side effects. Many patients were able to reduce their analgesic consumption. Quality of life improved and the majority of patients were happy with their treatment; in some cases, patients were able to return to work. Trial stimulation, which is relatively inexpensive and completely reversible, provides predictive value for long-term efficacy and increases the cost-effectiveness of permanent implantation. Studies consistently report that over time, SCS is potentially cost saving to the healthcare system. At present, SCS is considered a "last resort" in the treatment of refractory neuropathic pain, yet evidence suggests that early intervention with SCS results in greater efficacy and, in the case of FBSS, should be considered before re-operation.


Selenoprotein P and apolipoprotein E receptor-2 interact at the blood-brain barrier and also within the brain to maintain an essential selenium pool that protects against neurodegeneration.
Selenoprotein P (Sepp1) and its receptor, apolipoprotein E receptor 2 (apoER2), account for brain retaining selenium better than other tissues. The primary sources of Sepp1 in plasma and brain are hepatocytes and astrocytes, respectively. ApoER2 is expressed in varying amounts by tissues; within the brain it is expressed primarily by neurons. Knockout of Sepp1 or apoER2 lowers brain selenium from ∼120 to ∼50 ng/g and leads to severe neurodegeneration and death in mild selenium deficiency. Interactions of Sepp1 and apoER2 that protect against this injury have not been characterized. We studied Sepp1, apoER2, and brain selenium in knockout mice. Immunocytochemistry showed that apoER2 mediates Sepp1 uptake at the blood-brain barrier. When Sepp1(-/-) or apoER2(-/-) mice developed severe neurodegeneration caused by mild selenium deficiency, brain selenium was ∼35 ng/g. In extreme selenium deficiency, however, brain selenium of ∼12 ng/g was tolerated when both Sepp1 and apoER2 were intact in the brain. These findings indicate that tandem Sepp1-apoER2 interactions supply selenium for maintenance of brain neurons. One interaction is at the blood-brain barrier, and the other is within the brain. We postulate that Sepp1 inside the blood-brain barrier is taken up by neurons via apoER2, concentrating brain selenium in them.


High-Density Lipoproteins for Therapeutic Delivery Systems.
High-density lipoproteins (HDL) are a class of natural nanostructures found in the blood and are composed of lipids, proteins, and nucleic acids (e.g. microRNA). Their size, which appears to be well-suited for both tissue penetration/retention as well as payload delivery, long circulation half-life, avoidance of endosomal sequestration, and potential low toxicity are all excellent properties to model in a drug delivery vehicle. In this review, we consider high-density lipoproteins for therapeutic delivery systems. First we discuss the structure and function of natural HDL, describing in detail its biogenesis and transformation from immature, discoidal forms, to more mature, spherical forms. Next we consider features of HDL making them suitable vehicles for drug delivery. We then describe the use of natural HDL, discoidal HDL analogs, and spherical HDL analogs to deliver various classes of drugs, including small molecules, lipids, and oligonucleotides. We briefly consider the notion that the drug delivery vehicles themselves are therapeutic, constituting entities that exhibit "theralivery." Finally, we discuss challenges and future directions in the field.