Calcium transport by isolated skin of rainbow trout.
The skin overlying the cleithrum bone of freshwater-acclimated rainbow trout contains numerous mitochondria-rich (MR) cells, as detected by DASPEI fluorescence. This tissue was mounted in vitro in an Ussing-style chamber with fresh water on the mucosal surface and saline supplemented with bovine serum albumin on the serosal surface. The preparation developed a high transepithelial resistance and a small transepithelial potential (Vt), positive on the serosal side. Radioisotopic flux measurements indicated that the preparation actively transported Ca2+ from the mucosal to the serosal surface, as assessed by the Ussing flux ratio criterion. Ca2+ transport was positively correlated with MR cell density. Cortisol pretreatment in vivo reduced MR cell density and increased Vt but did not significantly alter Ca2+ fluxes. Ca2+ transport was unaffected by adrenergic agonists (10(-5) mol l-1 adrenaline, clonidine, isoprenaline) or cyclic AMP stimulants (10(-3) mol l-1 dibutyryl cyclic adenosine monophosphate, db-cAMP, plus 10(-4) mol l-1 isobutylmethylxanthine, IBMX) applied to the serosal surface. The Ca2+ ionophore ionomycin (1 x 10(-6)-3.2 x 10(-6) mol l-1 on the mucosal surface) increased both unidirectional Ca2+ fluxes and caused Ca2+ to accumulate within the epithelium. Lanthanum (10(-4) mol l-1) did not inhibit unidirectional Ca2+ fluxes, but apparently displaced Ca2+ from binding sites on the mucosal surface. Unlike Ca2+, movements of Na+ and Cl- across the epithelium were passive, as assessed by the flux ratio criterion, and neither adrenaline nor db-cAMP plus IBMX had any effect on Na+ or Cl- fluxes or electrical properties. These results indicate that ion transport across the skin mediated by MR cells ('chloride cells') contributes to Ca2+ but not to NaCl balance in freshwater trout.


Proclivity to elder abuse: a community study on Hong Kong Chinese.
This study aimed to provide preliminary estimates on proclivity to elder abuse and to determine the efficacy of the intergenerational transmission of violence and ecological theories in predicting elder abuse in contemporary Chinese societies. A total of 464 (225 males and 239 females) Chinese residing in Hong Kong completed questionnaires on attitudes toward elderly people, modernity, and filial piety as well as childhood experiences of abuse and proclivity to elder abuse. Results indicated that proclivity to verbal elder abuse was the most common among the three depicted types of abuse, accounting for 20% of the sample, whereas proclivity to physical and social elder abuse was less common, each accounting for 2.4%. A high level of childhood experience of abuse consistently emerged as the single most salient predictor for participants' endorsement of proclivity to elder abuse, while negative attitudes toward elderly people and modernity were the second and third most salient predictors.


A controlled trial of artemether or quinine in Vietnamese adults with severe falciparum malaria.
Artemisinin (qinghaosu) and its derivatives are rapidly effective antimalarial drugs derived from a Chinese plant. Preliminary studies suggest that these drugs may be more effective than quinine in the treatment of severe malaria. We studied artemether in Vietnam, where Plasmodium falciparum has reduced sensitivity to quinine. We conducted a randomized, double-blind trial in 560 adults with severe falciparum malaria. Two hundred seventy-six received intramuscular quinine dihydrochloride (20 mg per kilogram of body weight followed by 10 mg per kilogram every eight hours), and 284 received intramuscular artemether (4 mg per kilogram followed by 2 mg per kilogram every eight hours). Both drugs were given for a minimum of 72 hours. There were 36 deaths in the artemether group (13 percent) and 47 in the quinine group (17 percent; P = 0.16; relative risk of death in the patients given artemether, 0.74; 95 percent confidence interval, 0.5 to 1.11). The parasites were cleared more quickly from the blood in the artemether group (mean, 72 vs. 90 hours; P < 0.001); however, in this group fever resolved more slowly (127 vs. 90 hours, P < 0.001), the time to recovery from coma was longer (66 vs. 48 hours, P = 0.003), and the hospitalization was longer (288 vs. 240 hours, P = 0.005). Quinine treatment was associated with a higher risk of hypoglycemia (relative risk, 2.7; 95 percent confidence interval, 1.7 to 4.4; P < 0.001), but there were no other serious side effects in either group. Artemether is a satisfactory alternative to quinine for the treatment of severe malaria in adults.


Parental exposure to the herbicide diuron results in oxidative DNA damage to germinal cells of the Pacific oyster Crassostrea gigas.
Chemical pollution by pesticides has been identified as a possible contributing factor to the massive mortality outbreaks observed in Crassostrea gigas for several years. A previous study demonstrated the vertical transmission of DNA damage by subjecting oyster genitors to the herbicide diuron at environmental concentrations during gametogenesis. This trans-generational effect occurs through damage to genitor-exposed gametes, as measured by the comet-assay. The presence of DNA damage in gametes could be linked to the formation of DNA damage in other germ cells. In order to explore this question, the levels and cell distribution of the oxidized base lesion 8-oxodGuo were studied in the gonads of exposed genitors. High-performance liquid chromatography coupled with UV and electrochemical detection analysis showed an increase in 8-oxodGuo levels in both male and female gonads after exposure to diuron. Immunohistochemistry analysis showed the presence of 8-oxodGuo at all stages of male germ cells, from early to mature stages. Conversely, the oxidized base was only present in early germ cell stages in female gonads. These results indicate that male and female genitors underwent oxidative stress following exposure to diuron, resulting in DNA oxidation in both early germ cells and gametes, such as spermatozoa, which could explain the transmission of diuron-induced DNA damage to offspring. Furthermore, immunostaining of early germ cells seems indicates that damages caused by exposure to diuron on germ line not only affect the current sexual cycle but also could affect future gametogenesis.


Minimizing the Sample Sizes of Clinical Trials on Preclinical and Early Symptomatic Stage of Alzheimer Disease.
Clinical trials of investigational drugs for Alzheimer disease (AD) increasingly focus on the prodromal (symptomatic) stage of the illness and now its preclinical (asymptomatic) stage. Sensitive and specific cognitive and functional endpoints are needed to track subtle cognitive and functional changes in the early and preclinical stages to minimize sample sizes in these trials. To identify informative items in a standard clinical assessment protocol and a psychometric battery that are predictive of onset of dementia symptom. Longitudinal retrospective study. Washington University (WU) Knight Alzheimer Disease Research Center (ADRC). A total of 735 individuals at least 65 years old and cognitively normal at baseline from a longitudinal clinical cohort at the WU Knight ADRC. The annual clinical assessment included a wide spectrum of functional and cognitive domains; a comprehensive psychometric battery was completed about 2 weeks after the clinical evaluation. Psychometricians are blinded to the results of the clinical evaluation and to the prior performance of the participants on the psychometric tests. The mean age at baseline of the 735 participants was 74.30 and 62.31% were female. 240 individuals developed prodromal dementia symptoms (consistent with mild cognitive impairment due to AD and with very mild AD dementia) during longitudinal follow-up (mean follow-up=6.79 years). Among a total of 562 items in the clinical and cognitive assessments under analysis, 292 (52%) were identified as informative because their longitudinal changes were predictive of symptomatic onset. When these items were used to form the functional and cognitive composites, the longitudinal rates of changes were free of a learning effect and captured subtle longitudinal progression prior to symptomatic onset. The rates of change were much greater right after the symptomatic onset than those from the functional and cognitive composites formed using non-informative items. Although the sample sizes for prevention trials (prior to symptomatic onset) using the informative items still yield large numbers, the sample sizes for early treatment trial (after symptomatic onset) was much smaller than those derived from all the items or from the non-informative items alone. The antecedent longitudinal changes in nearly half of the items in a clinical assessment protocol and a comprehensive cognitive battery did not show statistically significant ability to predict the dementia symptom onset, and hence may be non-informative to track the preclinical functional and cognitive progression of AD. The remaining items, on the other hand, captured some of the preclinical changes prior to the symptom onset, but performed much better right after the symptom onset. Currently ongoing prevention trials on preclinical AD of elderly individuals may need to re-assess the sample sizes and statistical power.


DNA-based identification of forensically important Australian Sarcophagidae (Diptera).
The utility of the forensically important Sarcophagidae (Diptera) for time since death estimates has been severely limited, as morphological identification is difficult and thermobiological histories are inadequately documented. A molecular identification method involving the sequencing of a 658-bp 'barcode' fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene from 85 specimens, representing 16 Australian species from varying populations, was evaluated. Nucleotide sequence divergences were calculated using the Kimura-two-parameter distance model and a neighbour-joining phylogenetic tree generated. All species were resolved as reciprocally monophyletic, except Sarcophaga dux. Intraspecific and interspecific variation ranged from 0.000% to 1.499% (SE = 0.044%) and 6.658% to 8.983% (SE = 0.653%), respectively. The COI 'barcode' sequence was found to be suitable for the molecular identification of the studied Australian Sarcophagidae: 96.5% of the examined specimens were assigned to the correct species. Given that the sarcophagid fauna is poorly described, it is feasible that the few incorrectly assigned specimens represent cryptic species. The results of this research will be instrumental for implementation of the Australian Sarcophagidae in forensic entomology.


Characterisation of TGF-beta2 signalling and function in a human lens cell line.
There is increasing evidence implicating Transforming growth factor beta (TGF-beta) in pathological states of the lens. However, the underlying signalling mechanisms in human cells have not been fully examined. We have therefore investigated in a human lens cell line, FHL 124, the signalling characteristics of TGF-beta and Smad proteins. Moreover, we have tested the effectiveness of a fully human monoclonal anti-TGF-beta2 antibody, CAT-152, in suppressing TGF-beta2 induced changes in a number of conditions. FHL 124 cells were routinely cultured in Eagle's minimum essential medium (EMEM) supplemented with 10% FCS. Characterisation of the cell line was determined using Affymetrix gene microarrays and compared to native human lens epithelium. Cells were serum starved for 24 hr prior to exposure to TGF-beta2 in the presence and absence of CAT-152. Non-stimulated cells served as controls. Smad 4 localisation was observed by immunocytochemistry. To study Smad-dependent transcriptional activity, cells were transfected with SBE4-luc, an artificial smad-specific reporter, using Fugene-6. Transcriptional activity was determined by luciferase activity. Gene expression was assessed using reverse transcriptase-polymerase chain reaction (RT-PCR). Proliferation was determined by 3H-thymidine DNA incorporation. Growth and contraction were assessed using a scratch and patch assay. Affymettrix gene microarrays identified 99.5% homology between FHL 124 cells and the native lens epithelium with respect to expression pattern of the 22,270 genes on the chip. Moreover, FHL 124 cells expressed phenotypic markers, alphaA-crystallin and pax6 along with lens epithelial cell specific marker FoxE3. Immunocytochemical studies revealed the presence of Smad 4 which following TGF-beta2 exposure accumulated in the cell nucleus. Furthermore, Smad-dependent transcriptional activity was also stimulated. TGF-beta2 enhanced the expression of mRNA levels of alpha smooth muscle actin (alphaSMA) and connective tissue growth factor (CTGF). Exposure to TGF-beta2 resulted in a relatively small inhibition of 3H-thymidine incorporation of FHL 124 cells. However, a more marked contractile effect was also observed. In serum-supplemented medium, growth rates and TGF-beta induced contraction were enhanced. Treatment with 0.1-10 microg ml(-1) CAT-152 dose-dependently inhibited 10 ng ml(-1) TGF-beta2 induced effects in the presence and absence of serum. Exposure of FHL 124 cells to TGF-beta therefore induces Smad translocation, transcription, expression of transdifferentiation markers and induces marked contraction. Treatment with CAT-152 can effectively inhibit these responses. TGF-beta2 induced changes can also persist long after the period of exposure and when in the presence of serum TGF-beta induced contraction is enhanced. The work presented therefore demonstrates a platform technology to study TGF-beta2 signalling in human lens epithelial cells and provides evidence to show TGF-beta2 can be a potent factor in the development of posterior capsule opacification following cataract surgery.


Polycyclic aromatic hydrocarbons (PAHs) in sediments from a typical urban impacted river: application of a comprehensive risk assessment.
Soweto and Lenasia, the most densely populated area of South Africa, is simultaneously a thriving metropolis, with a fair share of people still living in squalor conditions directly dependant on the natural resources. Because of industrialisation the populace and environment in this urban area are exposed to various pollutants. The aquatic environment was selected as a proxy to study the effect of industrial pollution in this area. The concentrations, source identification, and various environmental risks of polycyclic aromatic hydrocarbons (PAHs) were determined in sediments of the upper reaches of the Klip River. Composite sediment samples collected in low-flow conditions in 2013 and 2014 ranged from 270-5400 ng/g. The PAHs in this aquatic ecosystem were dominated by 4-ring congeners and could be attributed to combustion of organic fuels by chemical mass balance. Heavy traffic and industrial complexes in the northern part of the study area were responsible for the PAH fingerprints. Probable adverse effects such as toxicity to benthic biota were proven after comparison with international sediment quality guidelines (SQG) both survey years. Toxic equivalence quotients (TEQs) calculated for the sediments using fish potency factors (FPFs) were up to 30 times greater than the Canadian guideline for dioxin-like compounds, indicating high probability of carcinogenic effect to fish mediated through the aryl-hydrocarbon receptor. Finally, sediments in the area posed moderate to high ecological risk, which corroborates the other toxicity assessments. The advantage of investigating multiple risk endpoints, is the comprehensive results obtained that allows for a more realistic representation of the study area. Consequently more aspects are kept into account that results in better conclusions.


Cell-type-specific enrichment of risk-associated regulatory elements at ovarian cancer susceptibility loci.
Understanding the regulatory landscape of the human genome is a central question in complex trait genetics. Most single-nucleotide polymorphisms (SNPs) associated with cancer risk lie in non-protein-coding regions, implicating regulatory DNA elements as functional targets of susceptibility variants. Here, we describe genome-wide annotation of regions of open chromatin and histone modification in fallopian tube and ovarian surface epithelial cells (FTSECs, OSECs), the debated cellular origins of high-grade serous ovarian cancers (HGSOCs) and in endometriosis epithelial cells (EECs), the likely precursor of clear cell ovarian carcinomas (CCOCs). The regulatory architecture of these cell types was compared with normal human mammary epithelial cells and LNCaP prostate cancer cells. We observed similar positional patterns of global enhancer signatures across the three different ovarian cancer precursor cell types, and evidence of tissue-specific regulatory signatures compared to non-gynecological cell types. We found significant enrichment for risk-associated SNPs intersecting regulatory biofeatures at 17 known HGSOC susceptibility loci in FTSECs (P = 3.8 × 10(-30)), OSECs (P = 2.4 × 10(-23)) and HMECs (P = 6.7 × 10(-15)) but not for EECs (P = 0.45) or LNCaP cells (P = 0.88). Hierarchical clustering of risk SNPs conditioned on the six different cell types indicates FTSECs and OSECs are highly related (96% of samples using multi-scale bootstrapping) suggesting both cell types may be precursors of HGSOC. These data represent the first description of regulatory catalogues of normal precursor cells for different ovarian cancer subtypes, and provide unique insights into the tissue specific regulatory variation with respect to the likely functional targets of germline genetic susceptibility variants for ovarian cancer.


Effect of aromatherapy with orange essential oil on salivary cortisol and pulse rate in children during dental treatment: A randomized controlled clinical trial.
Essential oils have been used as an alternative and complementary treatment in medicine. Citrus fragrance has been used by aromatherapists for the treatment of anxiety symptoms. Based on this claim, the aim of present study was to investigate the effect of aromatherapy with essential oil of orange on child anxiety during dental treatment. Thirty children (10 boys, 20 girls) aged 6-9 years participated in a crossover intervention study, according to the inclusion criteria, among patients who attended the pediatric department of Isfahan Dental School in 2011. Every child underwent two dental treatment appointments including dental prophylaxis and fissure-sealant therapy under orange aroma in one session (intervention) and without any aroma (control) in another one. Child anxiety level was measured using salivary cortisol and pulse rate before and after treatment in each visit. The data were analyzed using t-test by SPSS software version 18. The mean ± SD and mean difference of salivary cortisol levels and pulse rate were calculated in each group before and completion of treatment in each visit. The difference in means of salivary cortisol and pulse rate between treatment under orange odor and treatment without aroma was 1.047 ± 2.198 nmol/l and 6.73 ± 12.3 (in minutes), which was statistically significant using paired t-test (P = 0.014, P = 0.005, respectively). It seems that the use of aromatherapy with natural essential oil of orange could reduce salivary cortisol and pulse rate due to child anxiety state.


Differences Between New Immigrants From the Former Soviet Union and Veteran Residents in Knowledge, Perception, and Risk Factors of Stroke.
Stroke is the fourth most common cause of death in developed countries and a leading cause of acquired disability in adults. Awareness of risk factors and warning signs for stroke has a considerable impact on early arrival at the hospital and early thrombolytic treatment. Delays in seeking medical treatment following the onset of stroke symptoms have been shown to be more common among ethnic minorities. The aim of the current study was to examine stroke awareness and knowledge among new immigrants from the Former Soviet Union (IFSUs) compared with veteran residents (VRs). The study was conducted by students of the nursing master of arts program. Data were collected during March 2010 and June 2014. Trained registered nurses conducted interviews, using a structured, pretested, open-ended questionnaire. Participants were recruited by a snowball method from among the interviewers' friends and family members, 40 years or older with no history of stroke. A total of 643 Israelis, 420 VRs (65.3%) and 223 IFSUs (34.7%), were interviewed; 40.7% were men, with a mean age of 52.6 (SD, 9.3) years. Compared with VRs, IFSUs were more likely to report previous myocardial infarction (P = .022), hypertension (P < .001), and diabetes (P = .012). The mean number of stroke warning signs reported by IFSUs was higher than that reported by VRs (P = .031). When asked about risk factors for stroke, IFSUs more often stated hypertension (P = .03), whereas VRs more often reported family history (P = .03). Immigrants from the Former Soviet Union were more aware of strategies for the prevention of stroke than VRs (P = .02). The preferred sources of information about stroke for IFSUs, as distinguished from VRs, were personal doctors (P = .001) and radio programs (P = .03). Veteran residents showed lower levels of knowledge about stroke. Educational campaigns aimed at increasing knowledge of stroke among Israel's general population in Israel, as well as culturally targeting specific subgroups, are recommended.


In Vivo Antimalarial Activity of Annona muricata Leaf Extract in Mice Infected with Plasmodium berghei.
Malaria is one of the most important infectious diseases in the world. The choice for the treatment is highly limited due to drug resistance. Hence, finding the new compounds to treat malaria is urgently needed. The present study was attempted to evaluate the antimalarial activity of the Annona muricata aqueous leaf extract in Plasmodium berghei infected mice. Aqueous leaf extract of A. muricata was prepared and tested for acute toxicity in mice. For efficacy test in vivo, standard 4-day suppressive test was carried out. ICR mice were inoculated with 10(7) parasitized erythrocytes of P. berghei ANKA by intraperitoneal injection. The extracts (100, 500, and 1000 mg/kg) were then given orally by gavage once a day for 4 consecutive days. Parasitemia, percentage of inhibition, and packed cell volume were subsequently calculated. Chloroquine (10 mg/kg) was given to infected mice as positive control while untreated control was given only distilled water. It was found that A. muricata aqueous leaf extract at doses of 100, 500, and 1000 mg/kg resulted in dose dependent parasitemia inhibition of 38.03%, 75.25%, and 85.61%, respectively. Survival time was prolonged in infected mice treated with the extract. Moreover, no mortality to mice was observed with this extract up to a dose of 4000 mg/kg. In conclusion, the A. muricata aqueous leaf extract exerted significant antimalarial activity with no toxicity and prolonged survival time. Therefore, this extract might contain potential lead molecule for the development of a new drug for malaria treatment.


Clarification of the risk for venous thrombosis associated with hereditary protein S deficiency by investigation of a large kindred with a characterized gene defect.
Protein S is an important regulatory protein of the coagulation cascade. The risk for venous thrombosis associated with protein S deficiency has been uncertain because all previous risk estimates used phenotypic evaluation alone, which can be ambiguous. To quantitate the risk for thrombosis associated with a characterized protein S gene mutation that causes a Gly295-->Val substitution and protein S deficiency. Retrospective study of a single extended family. University hospital referral center. A 122-member protein S-deficient family, in which 44 members had a recently characterized gene defect. Comprehensive history of thrombosis, history of exposure to acquired risk factors for thrombosis, levels of total and free protein S antigen, and genotype for the mutation causing the Gly295-->Val substitution. Kaplan-Meier analysis of thrombosis-free survival showed that the probability of remaining free of thrombosis at 30 years of age is 0.5 (95% CI, 0.33 to 0.66) for carriers of the Gly295-->Val mutation compared with 0.97 (CI, 0.93 to 1.0) for normal family members (P < 0.001). In a multivariate Cox regression model that included smoking and obesity, the mutation was a strong independent risk factor for thrombosis (hazard ratio, 11.5 [CI, 4.33 to 30.6]; P < 0.001). For free (but not total) protein S antigen levels, the distributions of persons with and persons without the mutation did not overlap. Protein S deficiency, as defined by the presence of a causative gene mutation or a reduced level of free protein S antigen, is a strong independent risk factor for venous thrombosis in a clinical affected family.


Microspectrometer based on holographically recorded diffractive elements using supplementary holograms.
The conception and realization of an imaging microspectrometer, limited to an optical volume of 11 x 6 x 5 mm(3), is presented. The spectrometer is based on a multi-order concept and offers an overall spectral bandwidth of 400-1030 nm with better than 2.5 nm resolution in the visible range. The numerical aperture of NA = 0.2 allows an appropriate energy efficiency. As the most essential element of the microspectrometer, a concave diffraction grating with a diameter of 5 mm and an image distance of f = 8.6 mm was manufactured in a holographic recording process. For the recording process the specifications of the concave grating require two diffraction limited point sources in very close proximity. To provide a point source distance below 1 mm a recording concept based on the introduction of a supplementary hologram was employed.


Predictors of Adults' Knowledge and Awareness of HPV, HPV-Associated Cancers, and the HPV Vaccine: Implications for Health Education.
High human papillomavirus (HPV) prevalence and low HPV vaccine uptake are significant public health concerns. Disparities in HPV-associated cancers and HPV vaccine uptake rates suggest the need for additional research examining factors associated with vaccine acceptance. This study assessed HPV awareness and knowledge and identified sociodemographic characteristics associated with HPV knowledge at the population level. Data from adult men ( n = 1,197) and women ( n = 1,906) who participated in the National Cancer Institute's 2014 Health Information National Trends Survey were analyzed. Multivariable regression was used to identify predictors of four HPV knowledge categories: (1) general knowledge, (2) cervical cancer knowledge, (3) "other" cancer knowledge (i.e., anal, oral, penile), and (4) vaccine knowledge. Significant gender differences in awareness and knowledge of HPV and the HPV vaccine were revealed. Most participants (>70%) knew that HPV could cause cervical cancer, but fewer (14.9% to 31.5%) knew of the association between HPV and "other" cancers. Women were more likely to report that a health care provider recommended vaccination. Significant predictors of general HPV and HPV vaccine knowledge included gender, education, income, race, and other sociodemographic characteristics. Age and income predicted cervical cancer knowledge. Knowledge of "other" HPV-associated cancers was predicted by having a child under 18 years in the household and relationship status. HPV knowledge appears to be socially patterned. Low HPV knowledge among men and some racial minorities suggests a need for further intervention. Health education should emphasize risks of noncervical HPV-associated cancers. Patient-provider communication that includes education, counseling, and clear recommendations favoring vaccination may improve uptake.


Biotin dependency due to a defect in biotin transport.
We describe a 3-year-old boy with biotin dependency not caused by biotinidase, holocarboxylase synthetase, or nutritional biotin deficiency. We sought to define the mechanism of his biotin dependency. The child became acutely encephalopathic at age 18 months. Urinary organic acids indicated deficiency of several biotin-dependent carboxylases. Symptoms improved rapidly following biotin supplementation. Serum biotinidase activity and Biotinidase gene sequence were normal. Activities of biotin-dependent carboxylases in PBMCs and cultured skin fibroblasts were normal, excluding biotin holocarboxylase synthetase deficiency. Despite extracellular biotin sufficiency, biotin withdrawal caused recurrent abnormal organic aciduria, indicating intracellular biotin deficiency. Biotin uptake rates into fresh PBMCs from the child and into his PBMCs transformed with Epstein Barr virus were about 10% of normal fresh and transformed control cells, respectively. For fresh and transformed PBMCs from his parents, biotin uptake rates were consistent with heterozygosity for an autosomal recessive genetic defect. Increased biotin breakdown was ruled out, as were artifacts of biotin supplementation and generalized defects in membrane permeability for biotin. These results provide evidence for a novel genetic defect in biotin transport. This child is the first known with this defect, which should now be included in the identified causes of biotin dependency.


TolA modulates the oligomeric status of YbgF in the bacterial periplasm.
The trans-envelope Tol complex of Gram-negative bacteria is recruited to the septation apparatus during cell division where it is involved in stabilizing the outer membrane. The last gene in the tol operon, ybgF, is highly conserved, yet does not seem to be required for Tol function. We have addressed this anomaly by characterizing YbgF from Escherichia coli and its interaction with TolA, which, based on previous yeast two-hybrid data, is the only known physical link between YbgF and the Tol system. We show that the stable YbgF trimer undergoes a marked change in oligomeric state on binding TolA, forming a one-to-one complex with the Tol protein. Through a combination of pull-down assays, deletion analysis, and isothermal titration calorimetry, we map the TolA-YbgF interface to the C-terminal tetratricopeptide repeat domain of YbgF and 31 residues at the C-terminal end of TolA domain II (TolA(280-313)). We show that TolB, which binds TolA domain III close to the YbgF binding site, has no impact on the YbgF-TolA association. We also report the crystal structures of the two component domains of YbgF, the N-terminal coiled coil from E. coli YbgF, which forms a stable trimer and controls the oligomeric status of YbgF, and the monomeric tetratricopeptide repeat domain from Xanthomonas campestris YbgF, which is also able to trimerize. Although the coiled coil is not directly involved in TolA binding, we demonstrate that the regular hydrophilic patterning of its otherwise hydrophobic core is a prerequisite for the TolA-induced oligomeric-state transition of YbgF. We postulate that rather than YbgF affecting Tol function, it is the change in YbgF oligomeric status (with an accompanying change in its function) that likely explains the necessity for tight co-regulation of the ybgF and tol genes in Gram-negative bacteria.


Sparse model-based estimation of functional dependence in high-dimensional field and spike multiscale networks.
Behavior is encoded across multiple scales of brain activity, from binary neuronal spikes to continuous fields including local field potentials (LFP). Multiscale models need to describe both the encoding of behavior and the conditional dependencies in simultaneously recorded spike and field signals, which form a high-dimensional multiscale network. However, learning spike-field dependencies in high-dimensional recordings is challenging due to the prohibitively large number of spike-field signal pairs, which makes standard learning techniques subject to overfitting. We present a sparse model-based estimation algorithm to learn these multiscale network dependencies. We develop a multiscale encoding model consisting of a point process model of binary spikes for each neuron whose firing rate is a function of the LFP network features and behavioral states. Doing so, spike-field dependencies constitute the model parameters to be learned. We resolve the parameter learning challenge by forming a constrained optimization problem to maximize the likelihood with an L1 penalty term that eases the detection of significant spike-LFP dependencies. We then apply the Akaike information criterion (AIC) to force a sparse number of nonzero dependency parameters in the model. We validate the algorithm using simulations and spike-field data from two non-human primates (NHP) in a 3D motor task with motor cortical recordings and a pro-saccade visual task with prefrontal recordings. We find that by identifying a model with a sparse set of dependency parameters, the algorithm improves spike prediction compared with models without dependencies. Further, the algorithm identifies significantly fewer dependency parameters compared with standard methods while improving their spike prediction likely due to detecting fewer spurious dependencies. Also, spike prediction on any electrode improves by including LFP features from all electrodes compared with using only those on the same electrode. Finally, unlike standard methods, the algorithm uncovers patterns of spike-field network dependencies as a function of distance, brain region, and frequency band. This algorithm can help study functional dependencies in high-dimensional spike-field networks and leads to more accurate multiscale encoding models.


Two-dimensional imaging through diffusing media using 150-fs gated electronic holography techniques.
We combine 150-fs holographic gating with specifically adapted electronic holography techniques to produce transmission images of objects embedded in diffusing material. The technique captures, without the use of scanning procedures, complete two-dimensional images with 150-fs temporal resolution, thereby yielding submillimeter spatial resolution through diffusing objects several centimeters thick.


Hi! How are you? Response shift, implicit theories and differing epistemologies.
Measures of Health Related Quality of Life (HRQL) occupy a continuum, from highly standardized econometric methods such as the time tradeoff and standard gamble to individualized global measures. Each has its vocal adherents, each involves different assumptions about the nature and interpretation of HRQL, and each has potential advantages and disadvantages. In this paper, I begin by exploring two theories which attempt to explain how people make assessments of health over time: 'response shift' and the 'implicit theory of change' model. I show that the theories, which are based on different views of the underlying cognitive processes, make opposite predictions about the validity of prospective and retrospective judgments. I examine the broader issue of individualized vs. standardized questions, and discuss a fundamental epistemological difference which places the current discussion in a broader philosophical context. I propose that a partial resolution may arise from a more careful consideration of the goals of HRQL assessment in a particular situation.


Thermal comparison of novel bipolar forceps in bovine liver.
The goal of this work was to determine whether bipolar forceps using a novel heat pipe thermal regulation technology result in decreased thermal damage of untargeted collateral tissue compared with traditional bipolar forceps. Fresh ex vivo bovine livers underwent controlled coagulation with forceps with (n=36) or without (n=36) heat pipe technology. Liver specimens were assessed regarding the extent of thermal injury (heat pipe, n=20; non-heat pipe, n=20). During coagulation, tissue temperatures were measured via thermocouple array thermometry and imaged via infrared camera thermography (heat pipe, n=16; non-heat pipe, n=16). Forceps using heat pipe technology were associated with less thermal spread and demonstrated mean tissue temperatures 25% lower than observed with non-heat pipe forceps. The mean width, area, and depth of thermal injury were significantly reduced with heat pipe vs traditional forceps. In an ex vivo study of bovine liver bipolar coagulation, forceps that incorporated heat pipe technology limited thermal spread and reduced the extent of unintended injury to untargeted collateral tissue.


Circulating activated neutrophils in COVID-19: An independent predictor for mechanical ventilation and death.
Critical illness in COVID-19 is attributed to an exaggerated host immune response. Since neutrophils are the major component of innate immunity, we hypothesize that the quantum of activated neutrophils in the blood may predict an adverse outcome. In a retrospective study of 300 adult patients with confirmed COVID-19, we analyzed the impact of neutrophil activation (NEUT-RI), interleukin-6 (IL-6) and the established clinical risk factors of age, diabetes, obesity and hypertension on the clinical outcome. Significant predictors of the need for mechanical ventilation were NEUT-RI (Odds Ratio (OR) = 1.22, P < 0.001), diabetes (OR = 2.56, P = 0.00846) and obesity (OR = 6.55, P < 0.001). For death, the significant predictors were NEUT-RI (OR = 1.14, P = 0.00432), diabetes (OR = 4.11, P = 0.00185) and age (OR = 1.04, P = 0.00896). The optimal cut-off value for NEUT-RI to predict mechanical ventilation and death was 52 fluorescence intensity units (sensitivity 44%, specificity 88%, area under the curve 0.67 and 44%, 86%, 0.64, respectively). This finding supports an aberrant neutrophil response in COVID-19, likely due to uncontained viral replication, tissue hypoxia and exacerbated inflammation, introduces a novel biomarker for rapid monitoring and opens new avenues for therapeutic strategies.


Mechanical thrombectomy combined with recombinant tissue plasminogen activator thrombolysis in the venous sinus for the treatment of severe cerebral venous sinus thrombosis.
The aim of the present study was to assess the effectiveness and safety of endovascular interventional therapy, which is mechanical clot disruption combined with intrasinus thrombolytic therapy with recombinant tissue plasminogen activator (rt-PA), for severe cerebral venous sinus thrombosis (CVST). The records of eight patients with CVST confirmed by computed tomography, magnetic resonance imaging (MRI), magnetic resonance venography (MRV) and/or digital subtraction angiography were analyzed. Of the eight cases, the Glasgow Coma Scale (GCS) scores were between 4 and 9 with a mean value of 8.3±2.7. All the patients had venous infarction and two cases had intracranial hemorrhagic infarcts. Mechanical clot destruction combined with intrasinus thrombolytic therapy with rt-PA was performed under general anesthesia. Intravenous heparin therapy and intracerebral pressure control were applied during this period. One patient succumbed and the other seven patients showed good treatment efficacy. The GCS scores of the seven patients reverted to 15 upon discharge from the Northern Jiangsu People's Hospital (Yangzhou, China). With regard to the modified Rankin score of the seven patients three months following surgery, six patients scored 0 and one patient scored 1. MRI and MRV follow-up examinations were performed for 3-15 months. Complete recanalization of the criminal sinus, which refer to the sinus attributable to the infarction or hemorrhage, was observed in five cases and partial recanalization was observed in two cases. Symptoms were monitored for 3-24 months and no recurrence was observed. Therefore, mechanical thrombectomy combined with intrasinus thrombolytic therapy with rt-PA is safe and effective for patients with severe CVST.


A comparison between one- and two-fluoroscopic techniques in percutaneous vertebroplasty.
Percutaneous vertebroplasty (PV) is generally performed under fluoroscopic guidance. Technically, single fluoroscope is considered sufficient for effectively monitoring PV. However, single fluoroscopic technique might be time-consuming in rotating the C-arm of the fluoroscope for either antero-posterior (AP) or lateral radiographic view, and causing delay in detecting cement leakage that can occur if the correct sight is not given. The aim of the current investigation was to compare the efficacy and safety of performing PV using one or two sets of fluoroscope. This retrospective study enrolled 43 patients with painful osteoporotic vertebral fractures and they were treated with one-level PV. A single orthopaedic surgeon operated on all these patients. The patients were divided into two groups on the basis of the method of fluoroscopic control. In Group 1 (15 patients), PV was performed under the assistance of one fluoroscope. In Group 2 (28 patients), PV was performed under the control of two fluoroscopes. The mean follow-up was 19 months (range, 12 to 30). Neither symptomatic cement leakage nor postoperative infection was found in both groups. The mean operation time in Group 2 was shorter, 37.8 vs. 31.0 minutes for Groups 1 and 2, P = 0.03. The incidence of cement leakage for Groups 1 and 2 was 26.7% (4/15) vs. 14.3% (4/28), respectively, P = 0.19. We found that the two-fluoroscopic technique can provide simultaneous, real-time AP and lateral radiographic views to monitor entry point and cement delivery for PV and therefore reduce the operation time. The two-fluoroscopic technique did not require a complex manpower organization and has been proved to be a safe and effective technique for PV.


Ice ages and butterflyfishes: Phylogenomics elucidates the ecological and evolutionary history of reef fishes in an endemism hotspot.
For tropical marine species, hotspots of endemism occur in peripheral areas furthest from the center of diversity, but the evolutionary processes that lead to their origin remain elusive. We test several hypotheses related to the evolution of peripheral endemics by sequencing ultraconserved element (UCE) loci to produce a genome-scale phylogeny of 47 butterflyfish species (family Chaetodontidae) that includes all shallow water butterflyfish from the coastal waters of the Arabian Peninsula (i.e., Red Sea to Arabian Gulf) and their close relatives. Bayesian tree building methods produced a well-resolved phylogeny that elucidated the origins of butterflyfishes in this hotspots of endemism. We show that UCEs, often used to resolve deep evolutionary relationships, represent an important tool to assess the mechanisms underlying recently diverged taxa. Our analyses indicate that unique environmental conditions in the coastal waters of the Arabian Peninsula probably contributed to the formation of endemic butterflyfishes. Older endemic species are also associated with narrow versus broad depth ranges, suggesting that adaptation to deeper coral reefs in this region occurred only recently (<1.75 Ma). Even though deep reef environments were drastically reduced during the extreme low sea level stands of glacial ages, shallow reefs persisted, and as such there was no evidence supporting mass extirpation of fauna in this region.


Serotonin protects mouse liver from cholestatic injury by decreasing bile salt pool after bile duct ligation.
Obstructive cholestasis induces liver injury, postoperative complications, and mortality after surgery. Adaptive control of cholestasis, including bile salt homeostasis, is necessary for recovery and survival. Peripheral serotonin is a cytoprotective neurotransmitter also associated with liver regeneration. The effect of serotonin on cholestatic liver injury is not known. Therefore, we tested whether serotonin affects the severity of cholestatic liver injury. We induced cholestasis by ligation of the bile duct (BDL) in either wild-type (WT) mice or mice lacking peripheral serotonin (Tph1(-/-) and immune thrombocytopenic [ITP] mice). Liver injury was assessed by the levels of plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT) and tissue necrosis. Bile salt-regulating genes were measured by quantitative polymerase chain reaction and confirmed by western blotting and immunohistochemistry. Tph1(-/-) mice displayed higher levels of plasma AST, ALT, bile salts, and hepatic necrosis after 3 days of BDL than WT mice. Likewise, liver injury was disproportional in ITP mice. Moreover, severe cholestatic complications and mortality after prolonged BDL were increased in Tph1(-/-) mice. Despite the elevation in toxic bile salts, expression of genes involved in bile salt homeostasis and detoxification were not affected in Tph1(-/-) livers. In contrast, the bile salt reabsorption transporters Ostα and Ostβ were up-regulated in the kidneys of Tph1(-/-) mice, along with a decrease in urinary bile salt excretion. Serotonin reloading of Tph1(-/-) mice reversed this phenotype, resulting in a reduction of circulating bile salts and liver injury. We propose a physiological function of serotonin is to ameliorate liver injury and stabilize the bile salt pool through adaptation of renal transporters in cholestasis.


Eosinophilic esophagitis in children following cardiac transplantation: association with post-transplant lymphoproliferative disorder and other transplant outcomes.
Although cardiac transplantation is life-saving, morbidities from immunosuppression are significant. EoE is a complication of calcineurin inhibitors following liver transplant causing feeding intolerance, weight loss, vomiting, and dysphagia. There are limited reports of EoE following heart transplantation. We performed a retrospective single-center review of pediatric cardiac transplant patients from 2000 to 2010. A case-control analysis of patients with and without EoE was performed evaluating heart transplantation outcomes such as rates of rejection, CAV, PTLD, and graft loss. Eighty-six transplants were performed in 84 patients; 34 (40%) underwent diagnostic endoscopy, and 10 (12%) had EoE. Median time to diagnosis of EoE was 3.7 yr (IQR: 2.0-5.2). There were no differences in demographics or use of induction medications between patients with or without EoE. Patients with EoE had fewer episodes of treated rejection (1.0 vs. 2.5; p = 0.04). Four of 10 (40%) EoE patients had PTLD compared with only 2/24 (8%) of those without EoE (p = 0.048; OR 7.33 [95% CI: 1.1-50.2]). There were no differences in CAV or graft loss between groups. EoE should be considered as a cause of GI symptoms in children after cardiac transplantation and may be associated with fewer rejection episodes and increased rates of PTLD, thus representing a marker of over-immunosuppression.


Mechanistic Analysis of Fluorescence Quenching of Reduced Nicotinamide Adenine Dinucleotide by Oxamate in Lactate Dehydrogenase Ternary Complexes.
Fluorescence of Reduced Nicotinamide Adenine Dinucleotide (NADH) is extensively employed in studies of oxidoreductases. A substantial amount of static and kinetic work has focused on the binding of pyruvate or substrate mimic oxamate to the binary complex of lactate dehydrogenase (LDH)-NADH where substantial fluorescence quenching is typically observed. However, the quenching mechanism is not well understood limiting structural interpretation. Based on time-dependent density functional theory (TDDFT) computations with cam-B3LYP functional in conjunction with the analysis of previous experimental results, we propose that bound oxamate acts as an electron acceptor in the quenching of fluorescence of NADH in the ternary complex, where a charge transfer (CT) state characterized by excitation from the highest occupied molecular orbital (HOMO) of the nicotinamide moiety of NADH to the lowest unoccupied molecular orbital (LUMO) of oxamate exists close to the locally excited (LE) state involving only the nicotinamide moiety. Efficient quenching in the encounter complex like in pig heart LDH requires that oxamate forms a salt bridge with Arg-171 and hydrogen bonds with His-195, Thr-246 and Asn-140. Further structural rearrangement and loop closure, which also brings about another hydrogen bond between oxamate and Arg-109, will increase the rate of fluorescence quenching as well.


Environmental factors in genitourinary development.
In the last century the world has experienced an increase in the use of industrial chemicals as well as possible increases in the prevalence of hypospadias and cryptorchidism. Because hormones regulate the fetal development of many organs, numerous investigations have explored the role of environmental factors in genitourinary growth. We summarize the data regarding endocrine disruptors in human genitourinary development. A PubMed literature search was performed for human studies from 2004 to 2009. Few data exist on environmental influences on the kidneys, ureters or bladder. Studies on the influence of pesticides, vegetarian diets, diethylstilbestrol, oral contraceptives and corticosteroids on hypospadias have yielded varied conclusions. Phthalates appear to increase the odds of hypospadias and anogenital distance. The testicular dysgenesis syndrome postulates that cryptorchidism, hypospadias, poor semen quality and testicular cancer share a common environmental origin. In utero exposure to diethylstilbestrol has been shown to increase the risk of testicular dysgenesis syndrome. However, to our knowledge no other environmental factor has been shown to cause testicular dysgenesis syndrome. Some industrial chemicals as well as the pesticide dichloro-diphenyl-trichloroethane have detrimental effects on semen quality. In cases of documented industrial accidents, chemical exposure has also decreased the male-to-female birth ratio. Data on chemical exposure are largely mixed and inconclusive. Studies of populations with high exposure rates due to industrial accidents or in utero exposure to diethylstilbestrol suggest that endocrine disruptors adversely affect genitourinary development.


The mouse Clock mutation behaves as an antimorph and maps within the W19H deletion, distal of Kit.
Clock is a semidominant mutation identified from an N-ethyl-N-nitrosourea mutagenesis screen in mice. Mice carrying the Clock mutation exhibit abnormalities of circadian behavior, including lengthening of endogenous period and loss of rhythmicity. To identify the gene affected by this mutation, we have generated a high-resolution genetic map (> 1800 meioses) of the Clock locus. We report that Clock is 0.7 cM distal of Kit on mouse chromosome 5. Mapping shows that Clock lies within the W19H deletion. Complementation analysis of different Clock and W19H compound genotypes indicates that the Clock mutation behaves as an antimorph. This antimorphic behavior of Clock strongly argues that Clock defines a gene centrally involved in the mammalian circadian system.


Automatic Detection and Classification of Unsafe Events During Power Wheelchair Use.
Using a powered wheelchair (PW) is a complex task requiring advanced perceptual and motor control skills. Unfortunately, PW incidents and accidents are not uncommon and their consequences can be serious. The objective of this paper is to develop technological tools that can be used to characterize a wheelchair user's driving behavior under various settings. In the experiments conducted, PWs are outfitted with a datalogging platform that records, in real-time, the 3-D acceleration of the PW. Data collection was conducted over 35 different activities, designed to capture a spectrum of PW driving events performed at different speeds (collisions with fixed or moving objects, rolling on incline plane, and rolling across multiple types obstacles). The data was processed using time-series analysis and data mining techniques, to automatically detect and identify the different events. We compared the classification accuracy using four different types of time-series features: 1) time-delay embeddings; 2) time-domain characterization; 3) frequency-domain features; and 4) wavelet transforms. In the analysis, we compared the classification accuracy obtained when distinguishing between safe and unsafe events during each of the 35 different activities. For the purposes of this study, unsafe events were defined as activities containing collisions against objects at different speed, and the remainder were defined as safe events. We were able to accurately detect 98% of unsafe events, with a low (12%) false positive rate, using only five examples of each activity. This proof-of-concept study shows that the proposed approach has the potential of capturing, based on limited input from embedded sensors, contextual information on PW use, and of automatically characterizing a user's PW driving behavior.


Bonding to caries affected dentine.
Dentine replacement materials are often placed over caries affected dentine (CAD). The aim of this study was to compare the bonding characteristics and interactions of selected hydraulic calcium silicate-based dentine replacement materials to CAD and sound dentine. Three hydraulic calcium silicate-based dentine replacement materials were assessed: Retro MTA, Biodentine and Theracal LC. Material characterization was done by scanning electron microscopy and X-ray diffraction analyses. Blocks of sound and CAD were prepared and standardized by Vickers microhardness testing. Half of the affected and sound dentine blocks were pretreated with 5.25% NaOCl prior to material placement. The materials were stored either for 1 week or 24 weeks in 37°C in fully saturated conditions. Shear bond strength was assessed at both time periods. Radiopacity of the interfacial dentine was also evaluated to assess the remineralization potential of the dentine replacement materials. The reaction of Theracal was slower than that of the water-based materials. The bond strengths of different materials did not differ after 1 week (P>0.05). The bond strength of Biodentine and Retro MTA increased over time but no change was observed for Theracal. NaOCl pre-treatment deteriorated the bond strength to sound dentine but improvement was observed in affected dentine. Radiopacity changes were observed after 24 weeks. Biodentine and Retro MTA showed better bonding to CAD. Pretreatment with NaOCl improved the bond strength of dentine replacement materials to CAD.


Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus.
A multilocus sequence typing (MLST) scheme has been developed for Staphylococcus aureus. The sequences of internal fragments of seven housekeeping genes were obtained for 155 S. aureus isolates from patients with community-acquired and hospital-acquired invasive disease in the Oxford, United Kingdom, area. Fifty-three different allelic profiles were identified, and 17 of these were represented by at least two isolates. The MLST scheme was highly discriminatory and was validated by showing that pairs of isolates with the same allelic profile produced very similar SmaI restriction fragment patterns by pulsed-field gel electrophoresis. All 22 isolates with the most prevalent allelic profile were methicillin-resistant S. aureus (MRSA) isolates and had allelic profiles identical to that of a reference strain of the epidemic MRSA clone 16 (EMRSA-16). Four MRSA isolates that were identical in allelic profile to the other major epidemic MRSA clone prevalent in British hospitals (clone EMRSA-15) were also identified. The majority of isolates (81%) were methicillin-susceptible S. aureus (MSSA) isolates, and seven MSSA clones included five or more isolates. Three of the MSSA clones included at least five isolates from patients with community-acquired invasive disease and may represent virulent clones with an increased ability to cause disease in otherwise healthy individuals. The most prevalent MSSA clone (17 isolates) was very closely related to EMRSA-16, and the success of the latter clone at causing disease in hospitals may be due to its emergence from a virulent MSSA clone that was already a major cause of invasive disease in both the community and hospital settings. MLST provides an unambiguous method for assigning MRSA and MSSA isolates to known clones or assigning them as novel clones via the Internet.


Influence of taste disorders on dietary behaviors in cancer patients under chemotherapy.
To determine the relationship between energy and nutrient consumption with chemosensory changes in cancer patients under chemotherapy. We carried out a cross-sectional study, enrolling 60 subjects. Cases were defined as patients with cancer diagnosis after their second chemotherapy cycle (n = 30), and controls were subjects without cancer (n = 30). Subjective changes of taste during treatment were assessed. Food consumption habits were obtained with a food frequency questionnaire validated for Mexican population. Five different concentrations of three basic flavors --sweet (sucrose), bitter (urea), and a novel basic taste, umami (sodium glutamate)-- were used to measure detection thresholds and recognition thresholds (RT). We determine differences between energy and nutrient consumption in cases and controls and their association with taste DT and RT. No demographic differences were found between groups. Cases showed higher sweet DT (6.4 vs. 4.4 micromol/ml; p = 0.03) and a higher bitter RT (100 vs. 95 micromol/ml; p = 0.04) than controls. Cases with sweet DT above the median showed significant lower daily energy (2,043 vs.1,586 kcal; p = 0.02), proteins (81.4 vs. 54 g/day; p = 0.01), carbohydrates (246 vs.192 g/day; p = 0.05), and zinc consumption (19 vs.11 mg/day; p = 0.01) compared to cases without sweet DT alteration. Cases with sweet DT and RT above median were associated with lower completion of energy requirements and consequent weight loss. There was no association between flavors DT or RT and nutrient ingestion in the control group. Changes of sweet DT and bitter RT in cancer patients under chemotherapy treatment were associated with lower energy and nutrient ingestion. Taste detection and recognition thresholds disorders could be important factors in malnutrition development on patients with cancer under chemotherapy treatment.


Perceptual Competition Promotes Suppression of Reward Salience in Behavioral Selection and Neural Representation.
Visual attentional selection is influenced by the value of objects. Previous studies have demonstrated that reward-associated items lead to rapid distraction and associated behavioral costs, which are difficult to override with top-down control. However, it has not been determined whether a perceptually competitive environment could render the reward-driven distraction more susceptible to top-down suppression. Here, we trained both genders of human subjects to associate two orientations with high and low magnitudes of reward. After training, we collected fMRI data while the subjects performed a categorical visual search task. The item in the reward-associated orientation served as the distractor, and the relative physical salience between the target and distractor was carefully controlled to modulate the degree of perceptual competition. The behavioral results showed faster searches in the presence of high, relative to low, reward-associated distractors. However, this effect was evident only if the physical salience of the distractor was higher than that of the target, indicating a context-dependent suppression effect of reward salience that relied on high perceptual competition. By analyzing the fMRI data in primary visual cortex, we found that the behavioral pattern of results could be predicted by the suppressed channel responses tuned to the reward-associated orientation in the distractor location, accompanied by increased responses in the midbrain dopaminergic region. Our results suggest that the learned salience of a reward plays a flexible role in solving perceptual competition, enabling the neural system to adaptively modulate the perceptual representation for behavioral optimization.SIGNIFICANCE STATEMENT The predictiveness principle in learning theory suggests that the stimulus with high predictability of reward receives priority in attentional selection. This selection bias leads to difficulties in changing approach behaviors, and thus becomes an important factor related to psychiatric disorders with attentional deficits. Here, we demonstrated that such principle is adaptively implemented in attentional suppression in visual search. We showed that the learned salience induced the suppression of the reward-associated distractor if its competition with the target was strong and could not be readily solved. This behavioral pattern was accompanied by increased midbrain fMRI activity and weakened sensory representation of the reward-associated distractor in V1. Our findings provided direct evidence that our brain flexibly uses learned regularities in attentional control.


Cell death and hepatocarcinogenesis: Dysregulation of apoptosis signaling pathways.
Hepatocellular carcinoma (HCC) remains a disease with a poor prognosis despite recent advances in the pathophysiology and treatment. Although the disease is biologically heterogeneous, dysregulation of cellular proliferation and apoptosis both occur frequently and contribute to the malignant phenotype. Chronic liver disease is associated with intrahepatic inflammation which promotes dysregulation of cellular signaling pathways; this triggers proliferation and thus lays the ground for expansion of premalignant cells. Cancer emerges when immunological control fails and transformed cells develop resistance against cell death signaling pathways. The same mechanisms underlie the poor responsiveness of HCC towards chemotherapy. Only recently advances in understanding the signaling pathways involved has led to the development of an effective pharmacological therapy for advanced disease. The current review will discuss apoptosis signaling pathways and focus on apoptosis resistance of HCC involving derangements in cell death receptors (e.g. tumor necrosis factor-alpha [TNF], CD95/Apo-1, TNF-related apoptosis-inducing ligand [TRAIL]) and associated adapter molecules (e.g. FADD and FLIP) of apoptotic signaling pathways. In addition, the role of the transcription factor nuclear factor-kappaB (NFκB) and members of the B cell leukemia-2 (Bcl-2) family that contribute to the regulation of apoptosis in hepatocytes are discussed. Eventually, the delineation of cell death signaling pathways could contribute to the implementation of new therapeutic strategies to treat HCC.


Nitrogen-fixation activity and the abundance and taxonomy of nifH genes in agricultural, pristine, and urban prairie stream sediments chronically exposed to different levels of nitrogen loading.
Small streams exert great influences on the retention and attenuation of nitrogen (N) within stream networks. Human land use can lead to increased transport of dissolved inorganic N compounds and downstream eutrophication. Microbial activity in streams is important for maintaining an actively functioning N cycle. Chronically high N loading in streams affects the rates of the central processes of the N cycle by increasing rates of nitrification and denitrification, with biota exhibiting decreased efficiency of N use. The LINXII project measured N-cycle parameters in small streams using 15NO3- tracer release experiments. We concurrently measured N2 fixation rates in six streams of three types (agricultural, pristine, and urban prairie streams) as part of this broader study of major N-cycle processes. Nitrogen fixation in streams was significantly negatively correlated with nitrate levels, dissolved inorganic N levels, and denitrification rates. Algal mat and leaf litter samples generally exhibited the highest rates of N2 fixation. The abundance of nifH genes, as measured by real-time PCR, was marginally correlated with N2-fixation rates, but not to other N-cycle processes or stream characteristics. The nifH sequences observed were assigned to cyanobacteria, Deltaproteobacteria, Methylococcus, and Rhizobia. Seasonal changes, disturbances, and varying inputs may encourage a diverse, flexible, stable N2-fixing guild. Patchiness in the streams should be considered when assessing the overall impact of N2 fixation, since algal biomass exhibited high rates of N2 fixation.


The Relative Validity and Reproducibility of Food Frequency Questionnaires in the China Kadoorie Biobank Study.
Short versions of qualitative and quantitative food frequency questionnaires (FFQs) are widely used to assess usual food intake. However, fewer studies evaluated their relative validity and reproducibility in the Chinese population. This study compared 12-day 24-h dietary recalls with qualitative and quantitative FFQs designed by the China Kadoorie Biobank (CKB) study to assess the relative validity. Two FFQs were administered in the second and third seasons and compared to evaluate the reproducibility. Statistical tests included Spearman correlation coefficients, weighted kappa, and cross-classification. A total of 432 participants were eligible after stratifying by age, sex, and four regions. In the validation of qualitative FFQ, adjusted Spearman coefficients were between 0.23 and 0.59, and weighted kappa coefficients ranged from 0.61 to 0.88, except for fresh vegetables. The percentage of correct classification was highest in fresh vegetables and lowest in fresh fruit, but the percentages of extreme classification were below 3.0%. Corresponding Spearman and kappa coefficients for the reproducibility were 0.17-0.56 and 0.62-0.90. Furthermore, the correct classification constituted between 35.6 and 93.3% of all participants. Regarding the relative validity of the quantitative FFQ, Spearman coefficients ranged from 0.14 to 0.69 in addition to dried vegetables and carbonated soft drinks. For items with more than two-thirds of total participants consumed, weighted kappa coefficients were from 0.57 to 0.79; correct classification percentages were between 34.6% and 67.5%. Spearman and kappa coefficients for the reproducibility of the quantitative FFQ were 0.15-0.71 and 0.60-0.86, respectively; correct classification percentages varied from 47.8% to 71.6%. Most food items from the qualitative FFQ showed acceptable or even good relative validity and reproducibility in the CKB study. Likewise, major food items in the quantitative FFQ were valid and reproducible, but poor performances of dried vegetables and carbonated soft drinks indicated the need for modification and validation in future research.


Fibronectin tetrapeptide is target for syphilis spirochete cytadherence.
The syphilis bacterium, Treponema pallidum, parasitizes host cells through recognition of fibronectin (Fn) on cell surfaces. The active site of the Fn molecule has been identified as a four-amino acid sequence, arg-gly-asp-ser (RGDS), located on each monomer of the cell-binding domain. The synthetic heptapeptide gly-arg-gly-asp-ser-pro-cys (GRGDSPC), with the active site sequence RGDS, specifically competed with 125I-labeled cell-binding domain acquisition by T. pallidum. Additionally, the same heptapeptide with the RGDS sequence diminished treponemal attachment to HEp-2 and HT1080 cell monolayers. Related heptapeptides altered in one key amino acid within the RGDS sequence failed to inhibit Fn cell-binding domain acquisition or parasitism of host cells by T. pallidum. The data support the view that T. pallidum cytadherence of host cells is through recognition of the RGDS sequence also important for eukaryotic cell-Fn binding.


Cryo-EM Structure of the 2019-nCoV Spike in the Prefusion Conformation.
The outbreak of a novel betacoronavirus (2019-nCov) represents a pandemic threat that has been declared a public health emergency of international concern. The CoV spike (S) glycoprotein is a key target for urgently needed vaccines, therapeutic antibodies, and diagnostics. To facilitate medical countermeasure (MCM) development we determined a 3.5 Å-resolution cryo-EM structure of the 2019-nCoV S trimer in the prefusion conformation. The predominant state of the trimer has one of the three receptor-binding domains (RBDs) rotated up in a receptor-accessible conformation. We also show biophysical and structural evidence that the 2019-nCoV S binds ACE2 with higher affinity than SARS-CoV S. Additionally we tested several published SARS-CoV RBD-specific monoclonal antibodies and found that they do not have appreciable binding to nCoV-2019 S, suggesting antibody cross-reactivity may be limited between the two virus RBDs. The atomic-resolution structure of 2019-nCoV S should enable rapid development and evaluation of MCMs to address the ongoing public health crisis.


The constructal law of design and evolution in nature.
Constructal theory is the view that (i) the generation of images of design (pattern, rhythm) in nature is a phenomenon of physics and (ii) this phenomenon is covered by a principle (the constructal law): 'for a finite-size flow system to persist in time (to live) it must evolve such that it provides greater and greater access to the currents that flow through it'. This law is about the necessity of design to occur, and about the time direction of the phenomenon: the tape of the design evolution 'movie' runs such that existing configurations are replaced by globally easier flowing configurations. The constructal law has two useful sides: the prediction of natural phenomena and the strategic engineering of novel architectures, based on the constructal law, i.e. not by mimicking nature. We show that the emergence of scaling laws in inanimate (geophysical) flow systems is the same phenomenon as the emergence of allometric laws in animate (biological) flow systems. Examples are lung design, animal locomotion, vegetation, river basins, turbulent flow structure, self-lubrication and natural multi-scale porous media. This article outlines the place of the constructal law as a self-standing law in physics, which covers all the ad hoc (and contradictory) statements of optimality such as minimum entropy generation, maximum entropy generation, minimum flow resistance, maximum flow resistance, minimum time, minimum weight, uniform maximum stresses and characteristic organ sizes. Nature is configured to flow and move as a conglomerate of 'engine and brake' designs.


Infant regulatory function acts as a protective factor for later traits of autism spectrum disorder and attention deficit/hyperactivity disorder but not callous unemotional traits.
Reduced executive functions (EF) are commonly associated with developmental conditions (e.g., autism spectrum disorder, ASD; attention deficit/hyperactivity disorder, ADHD), although EF seems to be typical in children with callous unemotional (CU) traits. Regulatory function (RF) is a proposed infant precursor that maps on onto factors driving later EF. Here, we first test whether RF is specifically and negatively associated with ASD and ADHD traits, but not CU traits. Second, we test whether RF can act as a protective factor, by moderating the association between infant markers and subsequent ASD and ADHD traits. Participants were 79 infants at high (N = 42) and low (N = 37) familial risk for ASD. Data come from the 14-month infant visit (Autism Observational Scale for Infants; AOSI; activity level and RF from the Infant Behavior Questionnaire; IBQ) and the 7-year visit (ASD traits: Social Responsiveness Scale, SRS; ADHD traits: Conners 3, CU traits: Inventory of Callous Unemotional Traits). Infant RF was negatively associated with later traits of ASD (B = - 0.5, p = 0.01) and ADHD inattention (B = - 0.24, p = 0.02) but not hyperactivity (B = - 0.25, p = 0.10) or CU traits (B = 0.02, p = 0.86). RF moderated the association between infant AOSI score and ASD traits, with a significant effect in those with low RF (B = 0.10, p = 0.006), not high RF (B = 0.01, p = 0.78). Similarly, for ADHD, infant activity level was associated with later ADHD inattention in those with low (B = 0.17, p = 0.04) but not high RF (B = 0.07, p = 0.48). For ADHD hyperactivity symptoms, activity level was predictive at both high and low levels of RF. Strong RF may allow children to compensate for other atypicalities, thus attenuating the association between infant markers and later disorder traits. Whilst infant RF was associated with both ASD and ADHD inattention traits, there was no association with ADHD hyperactivity or CU traits. This suggests that any protective effect may not be universal and emphasises the need for a better understanding of the underlying moderating mechanisms.


Brain cortical activity is influenced by exercise mode and intensity.
Previous research suggests that different exercise modes and intensities lead to variations in brain cortical activity. However, because of variability and limitations in previous study designs, the mechanisms behind this link remain unclear. The aim of this study was to evaluate the effects on brain cortical activity that are dependent on exercise mode, intensity, and individual preferences and to localize these changes within the frontal, temporal, parietal, and occipital cortexes of the brain. Twelve runners were asked to perform four modes of exercise (treadmill, bicycle, arm crank, and isokinetic wrist flexions), each at 50% and 80% of their individual maximum capacity. In a second experiment, five hand cycling athletes completed an arm crank exercise protocol. Electrocortical activity was recorded for 5 min under rest conditions, before and after exercise. Standardized low-resolution brain electromagnetic tomography was used to localize changes in electroencephalographic α and β frequency ranges within the brain. We found effects of exercise mode and intensity. Moderate-intensity exercise (50% maximum) was followed by an increase in α activity in either somatosensory brain areas after "familiar exercise" or in emotional areas after "unfamiliar exercise" modes. After high-intensity exercise, changes in α and β frequencies were observed, which also seem to be specific to individual exercise familiarization/preference. When runners completed intense running and hand cyclists completed the intense arm crank exercise, we observed reductions of frontal β activity indicating a deactivation of emotional brain regions. In conclusion, these findings demonstrate that cortical activation patterns depend on exercise mode and intensity and that individual exercise preferences may contribute to the positive psychophysiological response.


Probing the ternary complexes of indoleamine and tryptophan 2,3-dioxygenases by cryoreduction EPR and ENDOR spectroscopy.
We have applied cryoreduction/EPR/ENDOR techniques to characterize the active-site structure of the ferrous-oxy complexes of human (hIDO) and Shewanella oneidensis (sIDO) indoleamine 2,3-dioxygenases, Xanthomonas campestris (XcTDO) tryptophan 2,3-dioxygenase, and the H55S variant of XcTDO in the absence and in the presence of the substrate L-Trp and a substrate analogue, L-Me-Trp. The results reveal the presence of multiple conformations of the binary ferrous-oxy species of the IDOs. In more populated conformers, most likely a water molecule is within hydrogen-bonding distance of the bound ligand, which favors protonation of a cryogenerated ferric peroxy species at 77 K. In contrast to the binary complexes, cryoreduction of all of the studied ternary [enzyme-O(2)-Trp] dioxygenase complexes generates a ferric peroxy heme species with very similar EPR and (1)H ENDOR spectra in which protonation of the basic peroxy ligand does not occur at 77 K. Parallel studies with L-Me-Trp, in which the proton of the indole nitrogen is replaced with a methyl group, eliminate the possibility that the indole NH group of the substrate acts as a hydrogen bond donor to the bound O(2), and we suggest instead that the ammonium group of the substrate hydrogen-bonds to the dioxygen ligand. The present data show that substrate binding, primarily through this H-bond, causes the bound dioxygen to adopt a new conformation, which presumably is oriented for insertion of O(2) into the C(2)-C(3) double bond of the substrate. This substrate interaction further helps control the reactivity of the heme-bound dioxygen by "shielding" it from water.


Is the abundance of Faecalibacterium prausnitzii relevant to Crohn's disease?
Reports that bacteria within the Firmicutes phylum, especially the species Faecalibacterium prausnitzii, are less abundant in Crohn's disease (CD) patients and supernatants from cultures of this bacterium are anti-inflammatory prompted the investigation of the possible correlations between the abundance of F. prausnitzii and the response to treatment in patients with gut diseases and healthy controls. In a randomized, double-blind trial, faeces were collected from healthy volunteers, and from patients with active CD, ulcerative colitis (UC) and irritable bowel syndrome before and after treatment. The levels of F. prausnitzii DNA in faecal suspensions were determined by PCR. Treatment by an elemental diet was effective, resulting in decreases in both the Harvey and Bradshaw index (P<0.001) and the concentrations of serum C-reactive protein (P<0.05). The total levels of F. prausnitzii in faecal samples from CD patients at presentation were lower than those in the other groups both before and after the treatment. There was no correlation between F. prausnitzii abundance and the severity of CD before treatment. Clinical improvement unexpectedly correlated with a significant decrease in the abundance of F. prausnitzii, especially the A2-165 subgroup (P<0.05). Our data suggest that a paucity of F. prausnitzii in the gastrointestinal microbial communities is likely to be a minor aetiological factor in CD: recovery following elemental diet is attributed to lower levels of gut flora.


Hypermagnesemia following an acute ingestion of Epsom salt in a patient with normal renal function.
Hypermagnesemia is a rare condition that commonly follows excessive therapeutic administration of magnesium sulfate to treat eclampsia of pregnancy. Signs and symptoms of this condition include extreme muscle weakness, loss of deep tendon reflexes, mental status depression, and cardiac dysrhythmias. Clinically significant hypermagnesemia following oral or rectal administration of magnesium containing products in patients with normal renal function is rare. We report a case of hypermagnesemia following massive Epsom salt ingestion that resulted in extreme musculoskeletal weakness and altered mentation.


Modeling the atomistic growth behavior of gold nanoparticles in solution.
The properties of gold nanoparticles strongly depend on their three-dimensional atomic structure, leading to an increased emphasis on controlling and predicting nanoparticle structural evolution during the synthesis process. In order to provide this atomistic-level insight and establish a link to the experimentally-observed growth behavior, a kinetic Monte Carlo simulation (KMC) approach is developed for capturing Au nanoparticle growth characteristics. The advantage of this approach is that, compared to traditional molecular dynamics simulations, the atomistic nanoparticle structural evolution can be tracked on time scales that approach the actual experiments. This has enabled several different comparisons against experimental benchmarks, and it has helped transition the KMC simulations from a hypothetical toy model into a more experimentally-relevant test-bed. The model is initially parameterized by performing a series of automated comparisons of Au nanoparticle growth curves versus the experimental observations, and then the refined model allows for detailed structural analysis of the nanoparticle growth behavior. Although the Au nanoparticles are roughly spherical, the maximum/minimum dimensions deviate from the average by approximately 12.5%, which is consistent with the corresponding experiments. Also, a surface texture analysis highlights the changes in the surface structure as a function of time. While the nanoparticles show similar surface structures throughout the growth process, there can be some significant differences during the initial growth at different synthesis conditions.


Beta-lactoglobulin assembles into amyloid through sequential aggregated intermediates.
We have investigated the aggregation and amyloid fibril formation of bovine beta-lactoglobulin variant A, with a focus on the early stages of aggregation. We used noncovalent labeling with thioflavin T and 1-anilino-8-naphthalenesulfonate to follow the conformational changes occurring in beta-lactoglobulin during aggregation using time resolved luminescence. 1-Anilino-8-naphthalenesulfonate monitored the involvement of the hydrophobic core/calyx of beta-lactoglobulin in the aggregation process. Thioflavin T luminescence monitored the formation of amyloid. The luminescence lifetime distributions of both probes showed changes that could be attributed to conformational changes occurring during and following aggregation. To correlate the luminescence measurements with the degree of aggregation and the morphology of the aggregates, we also measured dynamic light scattering and atomic force microscopy images. We evaluated the relative stability of the intermediates with an assay that is sensitive to aggregation reversibility. Our results suggest that initial aggregation during the first 5 days occurred with partial disruption of the characteristic calyx in beta-lactoglobulin. As the globular aggregates grew from days 5 to 16, the calyx was completely disrupted and the globular aggregates became more stable. After this second phase of aggregation, conversion into a fibrillar form occurred, marking the growth phase, and still more changes in the luminescence signals were observed. Based on these observations, we propose a three-step process by which monomer is converted first into weakly associated aggregates, which rearrange into stable aggregates, which eventually convert into protofibrils that elongate in the growth phase.


A dodecapeptide (YQVTQSKVMSHR) exhibits antibacterial effect and induces cell aggregation in Escherichia coli.
Antimicrobial peptides play an important role in the innate immune response and host defense mechanism. In the present study, we employed phage display technique to screen for inhibitors which may block the phosphoenolpyruvatedependent phosphotransferase system (PTS) pathway and hence retard cell growth. The recombinant histidine-containing phosphocarrier HPr protein was prepared as the target to screen for the tight binders from the phage-displayed random peptide library Ph.D.-12. The biopanning processes were performed and the binding capabilities of the selected phage were further estimated by enzyme-linked immunosorbent assay (ELISA). The single-stranded DNAs of the 20 selected phages were isolated, sequenced, and five corresponding peptides were synthesized. Only one of the five peptides, AP1 (YQVTQSK VMSHR) was found to inhibit the growth of Escherichia coli cells efficiently (IC₅₀~50 μM). Molecular modeling reveals that AP1 may block the EI-HPr interaction and phosphotransfer. Interestingly, AP1 was also found to induce cell aggregation in a concentration-dependent manner. Since glycogen accumulation has been attributed to biofilm formation, the effects of AP1 on the intracellular glycogen levels were measured. The results strongly indicate that the cell aggregation may be caused by the binding of peptide AP1 with HPr to block the interaction of dephosphorylated HPr with glycogen phosphorylase (GP). Because glycogen phosphorylase activity can be activated by HPr-GP interaction, the binding of AP1 to HPr would cause a decreasing rate of glycogen breakdown in M9 medium and accumulation of glycogen, which may lead to eventual cell aggregation. To the best of our knowledge, this is the first study to demonstrate that an inhibitor bound to a dephosphorylated HPr can decouple its regulatory function and induce cell aggregation.


Use of rivastigmine or galantamine and risk of adverse cardiac events: a database study from the Netherlands.
Two cholinesterase inhibitors (ChEIs), rivastigmine and galantamine, are used to treat Alzheimer disease in the Netherlands. Several adverse cardiac events have been reported for these medications. We aimed to assess if the use of ChEIs increased the risk of cardiac events in the Netherlands. A cohort crossover study of the PHARMO Record Linking System database included patients who initiated ChEIs at age 50 years or older, had at least 1 dispensing of a ChEI drug between 1998 and 2008, a 1-year history in PHARMO, and 1 subsequent dispensing of any medication. Two outcomes were assessed: a first hospitalization for syncope or atrioventricular block. Poisson and Cox regression were used to calculate incidence densities and hazard ratios for cardiac events during periods with ChEI use, compared with periods without ChEI use. During the complete observation period of 8.9 years (interquartile range 6.7 to 10.2) there were 132 first hospitalizations for atrioventricular block and 17 first hospitalizations for syncope among 3358 patients. The adjusted incidence densities were significantly increased during ChEI exposure for syncope and atrioventricular block, when compared with the background incidence densities in the roughly 5 years before the last year before ChEI initiation. However, when exposed periods were compared with the unexposed periods 1 year before ChEI initiation and times after exposure, the adjusted hazard ratios remained increased for syncope and atrioventricular block, but increases were not significant anymore. Exposure to ChEIs might increase the risk of adverse cardiac events, but small numbers of cases limit conclusions about the risk in this population and research on larger study samples is needed.


Immunoregulation of follicular renewal, selection, POF, and menopause in vivo, vs. neo-oogenesis in vitro, POF and ovarian infertility treatment, and a clinical trial.
The immune system plays an important role in the regulation of tissue homeostasis ("tissue immune physiology"). Function of distinct tissues during adulthood, including the ovary, requires (1) Renewal from stem cells, (2) Preservation of tissue-specific cells in a proper differentiated state, which differs among distinct tissues, and (3) Regulation of tissue quantity. Such morphostasis can be executed by the tissue control system, consisting of immune system-related components, vascular pericytes, and autonomic innervation. Morphostasis is established epigenetically, during morphogenetic (developmental) immune adaptation, i.e., during the critical developmental period. Subsequently, the tissues are maintained in a state of differentiation reached during the adaptation by a "stop effect" of resident and self renewing monocyte-derived cells. The later normal tissue is programmed to emerge (e.g., late emergence of ovarian granulosa cells), the earlier its function ceases. Alteration of certain tissue differentiation during the critical developmental period causes persistent alteration of that tissue function, including premature ovarian failure (POF) and primary amenorrhea. In fetal and adult human ovaries the ovarian surface epithelium cells called ovarian stem cells (OSC) are bipotent stem cells for the formation of ovarian germ and granulosa cells. Recently termed oogonial stem cells are, in reality, not stem but already germ cells which have the ability to divide. Immune system-related cells and molecules accompany asymmetric division of OSC resulting in the emergence of secondary germ cells, symmetric division, and migration of secondary germ cells, formation of new granulosa cells and fetal and adult primordial follicles (follicular renewal), and selection and growth of primary/preantral, and dominant follicles. The number of selected follicles during each ovarian cycle is determined by autonomic innervation. Morphostasis is altered with advancing age, due to degenerative changes of the immune system. This causes cessation of oocyte and follicular renewal at 38 +/-2 years of age due to the lack of formation of new granulosa cells. Oocytes in primordial follicles persisting after the end of the prime reproductive period accumulate genetic alterations resulting in an exponentially growing incidence of fetal trisomies and other genetic abnormalities with advanced maternal age. The secondary germ cells also develop in the OSC cultures derived from POF and aging ovaries. In vitro conditions are free of immune mechanisms, which prevent neo-oogenesis in vivo. Such germ cells are capable of differentiating in vitro into functional oocytes. This may provide fresh oocytes and genetically related children to women lacking the ability to produce their own follicular oocytes. Further study of "immune physiology" may help us to better understand ovarian physiology and pathology, including ovarian infertility caused by POF or by a lack of ovarian follicles with functional oocytes in aging ovaries. The observations indicating involvement of immunoregulation in physiological neo-oogenesis and follicular renewal from OSC during the fetal and prime reproductive periods are reviewed as well as immune system and age-independent neo-oogenesis and oocyte maturation in OSC cultures, perimenopausal alteration of homeostasis causing disorders of many tissues, and the first OSC culture clinical trial.


Stress-Activated Kinase Mitogen-Activated Kinase Kinase-7 Governs Epigenetics of Cardiac Repolarization for Arrhythmia Prevention.
Ventricular arrhythmia is a leading cause of cardiac mortality. Most antiarrhythmics present paradoxical proarrhythmic side effects, culminating in a greater risk of sudden death. We describe a new regulatory mechanism linking mitogen-activated kinase kinase-7 deficiency with increased arrhythmia vulnerability in hypertrophied and failing hearts using mouse models harboring mitogen-activated kinase kinase-7 knockout or overexpression. The human relevance of this arrhythmogenic mechanism is evaluated in human-induced pluripotent stem cell-derived cardiomyocytes. Therapeutic potentials by targeting this mechanism are explored in the mouse models and human-induced pluripotent stem cell-derived cardiomyocytes. Mechanistically, hypertrophic stress dampens expression and phosphorylation of mitogen-activated kinase kinase-7. Such mitogen-activated kinase kinase-7 deficiency leaves histone deacetylase-2 unphosphorylated and filamin-A accumulated in the nucleus to form a complex with Krüppel-like factor-4. This complex leads to Krüppel-like factor-4 disassociation from the promoter regions of multiple key potassium channel genes (Kv4.2, KChIP2, Kv1.5, ERG1, and Kir6.2) and reduction of their transcript levels. Consequent repolarization delays result in ventricular arrhythmias. Therapeutically, targeting the repressive function of the Krüppel-like factor-4/histone deacetylase-2/filamin-A complex with the histone deacetylase-2 inhibitor valproic acid restores K+ channel expression and alleviates ventricular arrhythmias in pathologically remodeled hearts. Our findings unveil this new gene regulatory avenue as a new antiarrhythmic target where repurposing of the antiepileptic drug valproic acid as an antiarrhythmic is supported.


DNA hypermethylation and DNA hypomethylation is present at different loci in chronic kidney disease.
Genetic risk factors for chronic kidney disease (CKD) are being identified through international collaborations. By comparison, epigenetic risk factors for CKD have only recently been considered using population-based approaches. DNA methylation is a major epigenetic modification that is associated with complex diseases, so we investigated methylome-wide loci for association with CKD. A total of 485,577 unique features were evaluated in 255 individuals with CKD (cases) and 152 individuals without evidence of renal disease (controls). Following stringent quality control, raw data were quantile normalized and β values calculated to reflect the methylation status at each site. The difference in methylation status was evaluated between cases and controls with resultant P values adjusted for multiple testing. Genes with significantly increased and decreased levels of DNA methylation were considered for biological relevance by functional enrichment analysis using KEGG pathways in Partek Genomics Suite. Twenty-three genes, where more than one CpG per loci was identified with Padjusted<10(-8), demonstrated significant methylation changes associated with CKD and additional support for these associated loci was sought from published literature. Strong biological candidates for CKD that showed statistically significant differential methylation include CUX1, ELMO1, FKBP5, INHBA-AS1, PTPRN2, and PRKAG2 genes; several genes are differentially methylated in kidney tissue and RNA-seq supports a functional role for differential methylation in ELMO1 and PRKAG2 genes. This study reports the largest, most comprehensive, genome-wide quantitative evaluation of DNA methylation for association with CKD. Evidence confirming methylation sites influence development of CKD would stimulate research to identify epigenetic therapies that might be clinically useful for CKD.


A tool for self-assessment of communication skills and professionalism in residents.
Effective communication skills and professionalism are critical for physicians in order to provide optimum care and achieve better health outcomes. The aims of this study were to evaluate residents' self-assessment of their communication skills and professionalism in dealing with patients, and to evaluate the psychometric properties of a self-assessment questionnaire. A modified version of the American Board of Internal Medicine's (ABIM) Patient Assessment survey was completed by 130 residents in 23 surgical and non-surgical training programs affiliated with a single medical school. Descriptive, regression and factor analyses were performed. Internal consistency, inter-item gamma scores, and discriminative validity of the questionnaire were determined. Factor analysis suggested two groups of items: one group relating to developing interpersonal relationships with patients and one group relating to conveying medical information to patients. Cronbach's alpha (0.86) indicated internal consistency. Males rated themselves higher than females in items related to explaining things to patients. When compared to graduates of U.S. medical schools, graduates of medical schools outside the U.S. rated themselves higher in items related to listening to the patient, yet lower in using understandable language. Surgical residents rated themselves higher than non-surgical residents in explaining options to patients. This appears to be an internally consistent and reliable tool for residents' self-assessment of communication skills and professionalism. Some demographic differences in self-perceived communication skills were noted.


Evaluation of bis-cyclometalated alkynylgold(iii) sensitizers for water photoreduction to hydrogen.
Well-defined gold sensitizers for hydrogen production from water remain extremely rare despite decades of interest, and are currently limited to systems based on ruthenium, iridium or platinum complexes. This report details the synthesis and characterization of a series of neutral cyclometalated gold(iii) complexes of the type [(RC^N^CR)Au(C[triple bond, length as m-dash]C-R')] (R = H or tert-butyl group; R' = aryl groups) that have been found to be good candidates to function as harvesting materials in light-induced electron transfer reactions. We established the efficacy of systems with these gold(iii) complexes as photosensitizers (PSs) in the production of renewable hydrogen in the presence of [Co(2,2'-bipyridine)3]Cl2 or [Rh(4,4'-di-tert-butyl-2,2'-bipyridine)3](PF6)3 as a H2-evolved catalyst and triethanolamine (TEOA) as a sacrificial electron donor in acetone-water solution. All complexes are active, and there is a more than threefold increase over other candidates in photocatalytic H2 generation activity. Under the optimal reaction conditions, hydrogen evolution took place through a photochemical route with the highest efficiency and with a turnover number (TON) of up to 1441.5 relative to the sensitizer over 24 hours. In the initial photochemical path, the reductive quenching of the excited gold(iii) complex by TEOA due to the latter's greater concentration in the system followed by electron transfer to the catalyst species is proposed to be the dominant mechanism. A photo-to-H2 quantum yield of approximately 13.7% was attained when illuminated with monochromatic light of 400 nm. Such gold(iii) complexes have demonstrated significant utility in solar-to-hydrogen reactions and thus represent a new effective class of light-harvesting materials. These results open possibilities for pursuing more efficient photosensitizers featuring gold(iii) complexes in photocatalytic solar energy conversion.


Ambulatory blood pressure monitoring in diabetic hypertensive patients, single center report--preliminary results.
ABPM (ambulatory blood pressure monitoring) has been considered to be a useful tool for the diagnosis and management of arterial hypertension and is a better predictor of future cardiovascular events as compared with conventional office-based BP measurements. Despite its potential values, ABPM is not yet widely used in many clinical offices mainly because of lack of knowledge and unavailability. Aims of this preliminary study are to determine the control of hypertension and circadian BP characteristics in patients referred to our Centre whom we enrolled in the "HRKMAT" Study-Croatian Registry of ABPM. Although patients included in HRKMAT Study had other risk factors for cardiovascular diseases, in this paper we analyzed differences between hypertensive diabetics (N = 20) and nondiabetics (N = 57). 24-hours ABPM was performed with an automated oscillometric device Mobil-O-Graph NG Vers.20 and office BP using mercury sphygmomanometer. Average office BP was 139/90 mmHg, and average 24h ABPM was 130/82 mmHg. Majority of hypertensive patients used antihypertensive drugs (79.2%). Diabetic patients had higher systolic BP but lower diastolic BP. There were no statistically significant differences in dipping status, but earlier BP surge was noticed in reverse diabetic dippers than in reverse non-diabetic dippers. Though no significant, there was higher prevalence of WCH ("white coat hypertension") in diabetics, and we found MH (masked hypertension) in only two patients. These are preliminary results on ABPM from our centre and of HRKMAT registry. Further and more valuable data and results are awaited from the main HRKMAT database.