Labyrinthopeptins as virolytic inhibitors of respiratory syncytial virus cell entry.
Acute lower respiratory tract infections (ALRI) caused by respiratory syncytial virus (RSV) are associated with a severe disease burden among infants and elderly patients. Treatment options are limited. While numerous drug candidates with different viral targets are under development, the utility of RSV entry inhibitors is challenged by a low resistance barrier and by single mutations causing cross-resistance against a wide spectrum of fusion inhibitor chemotypes. We developed a cell-based screening assay for discovery of compounds inhibiting infection with primary RSV isolates. Using this system, we identified labyrinthopeptin A1 and A2 (Laby A1/A2), lantibiotics isolated from Actinomadura namibiensis, as effective RSV cell entry inhibitors with IC50s of 0.39 μM and 4.97 μM, respectively, and with favourable therapeutic index (>200 and > 20, respectively). Both molecules were active against multiple RSV strains including primary isolates and their antiviral activity against RSV was confirmed in primary human airway cells ex vivo and a murine model in vivo. Laby A1/A2 were antiviral in prophylactic and therapeutic treatment regimens and displayed synergistic activity when applied in combination with each other. Mechanistic studies showed that Laby A1/A2 exert virolytic activity likely by binding to phosphatidylethanolamine moieties within the viral membrane and by disrupting virus particle membrane integrity. Probably due to its specific mode of action, Laby A1/A2 antiviral activity was not affected by common resistance mutations to known RSV entry inhibitors. Taken together, Laby A1/A2 represent promising candidates for development as RSV inhibitors. Moreover, the cell-based screening system with primary RSV isolates described here should be useful to identify further antiviral agents.


Evaluation of preparedness for medical emergencies at dental offices: A survey.
The best way to handle an emergency is to be prepared in advance. The purpose of this study was to assess the availability of medical emergency drugs at dental offices and to determine the level of knowledge and preparedness of dentists to manage medical emergencies at their dental offices. A descriptive cross-sectional study was conducted from January to June 2014 with 250 dental graduates to determine their knowledge and ability in the management of medical emergencies and assess availability of emergency drugs and equipments in dental offices in the Ahmedabad and Udaipur areas of India. The questionnaire consisted of mainly objective questions, requiring a simple yes or no reply. Mann-Whitney test was used for the analysis. A 95% confidence level was used and a P value of less than or equal to 0.05 was considered statistically significant. The results of our study showed that almost all the surveyed dentists (98%) enquired about medical history, but only 12% obtained filled health history proforma from patients regarding the above. Only 38.4% participants recorded the vital signs of patients before commencing any treatment. Also, 7.6% reported about having attended any workshops on emergency training or management programs. Emergency kits were available with only 24% participants and the available kits were assessed for the availability of emergency drugs. Also, 34% were confident about giving intramuscular injection and only 6.6% were sure about giving intravenous injections. The P value was found to be highly statistically significant. The results of our study reflect an alarming situation about the capability of dentists to deal with such conditions.


People with impaired glucose tolerance and impaired fasting glucose are similarly susceptible to cardiovascular disease: a study in first-degree relatives of type 2 diabetic patients.
To compare the cardiovascular disease (CVD) risk factors between subjects with impaired fasting glucose (IFG) and those with impaired glucose tolerance (IGT) in the first-degree relatives (FDR) of type 2 diabetic patients. A cross-sectional study, conducted between 2004 and 2006 in 1,893 (1,412 females and 481 males) FDR of type 2 diabetic outpatients of the Isfahan Endocrine and Metabolism Research Center. In all participants, blood pressure, weight, height, waist circumference, serum lipids and HbA1c were measured and a standard 75-g 2-hour oral glucose tolerance test was performed. The diagnosis of IGT, IFG and diabetes was made according to American Diabetes Association criteria. Isolated IGT and isolated IFG, and both IFG and IGT were observed in 8.8%, 17.4% and 11.2% of subjects, respectively. In comparison to subjects with normal glucose levels and tolerance (control group): the mean waist circumference was significantly higher in both IFG and IGT groups; BMI, HDL-c and LDL-c in the IFG group; and triglycerides in the IGT group. The means of all studied CVD risk factors were significantly higher in the IGT+IFG group than the control group, except for blood pressure, HDL-c and HbA1c. No significant differences were found regarding CVD risk factors between IFG and IGT groups. The prevalence of IFG and IGT is high in FDR of type 2 diabetic patients. CVD risk factors are similar in these 2 groups and higher than in the control group. More attention should be paid to screening and treatment of this high-risk population.


The course of the epidemic of acquired immunodeficiency syndrome in the United States hemophilia population.
The time course of the epidemic of acquired immunodeficiency syndrome (AIDS) as it has occurred in the US hemophilia population is examined using surveillance data collected by the Centers for Disease Control (CDC). These data indicate that the epidemic course in hemophiliacs is distinguishable from that in the homosexual/bisexual and intravenous drug-using populations in at least one respect--the epidemic in the hemophilia population is characterized by a lack of consistent increase in the number of new AIDS cases in successive time intervals. This difference is interpreted as being attributable to the mechanisms by which AIDS virus is spread among hemophiliacs. In addition, the short survival following diagnosis of AIDS in hemophiliacs and the magnitude of yearly incidence rates for this group in 1984 and 1985 show the hemophilia population to have been severely affected by the epidemic.


Immunoglobulin transcript sequence and somatic hypermutation computation from unselected RNA-seq reads in chronic lymphocytic leukemia.
Immunoglobulins (Ig) are produced by B lymphocytes as secreted antibodies or as part of the B-cell receptor. There is tremendous diversity of potential Ig transcripts (>1 × 10(12)) as a result of hundreds of germ-line gene segments, random nucleotide incorporation during joining of gene segments into a complete transcript, and the process of somatic hypermutation at individual nucleotides. This recombination and mutation process takes place in the maturing B cell and is responsible for the diversity of potential epitope recognition. Cancers arising from mature B cells are characterized by clonal production of Ig heavy (IGH@) and light chain transcripts, although whether the sequence has undergone somatic hypermutation is dependent on the maturation stage at which the neoplastic clone arose. Chronic lymphocytic leukemia (CLL) is the most common leukemia in adults and arises from a mature B cell with either mutated or unmutated IGH@ transcripts, the latter having worse prognosis and the assessment of which is routinely performed in the clinic. Currently, IGHV mutation status is assessed by Sanger sequencing and comparing the transcript to known germ-line genes. In this paper, we demonstrate that complete IGH@ V-D-J sequences can be computed from unselected RNA-seq reads with results equal or superior to the clinical procedure: in the only discordant case, the clinical transcript was out-of-frame. Therefore, a single RNA-seq assay can simultaneously yield gene expression profile, SNP and mutation information, as well as IGHV mutation status, and may one day be performed as a general test to capture multidimensional clinically relevant data in CLL.


Spike-timing relationship of neurochemically-identified dorsal raphe neurons during cortical slow oscillations.
The firing activity of dorsal raphe neurons is related to arousal state. However, it is unclear how this firing activity is precisely related to cortical activity, in particular oscillations occurring during sleep rhythms. Here we conducted single-cell extracellular recordings and juxtacellular labelling while monitoring electrocorticogram (ECoG) activity in urethane anaesthetised rats, to relate activity in neurochemically identified groups of neurons to cortical slow-wave activity (SWA). We observed that electrophysiological heterogeneity in dorsal raphe neurons revealed different neurochemical groups of DRN neurons and was mirrored by significant differences in the phase and strength of coupling to the cortical slow oscillations. Spike firing relationship of clock-like neurons, identified as 5-HT (5-hydroxytryptamine) or serotonin neurons, was higher during the inactive component of the oscillations. In contrast, half of the identified bursting 5-HT neurons did not exhibit strong cortical entrainment; those that did fired most during the inactive component of the SWA. Two groups of putatively non-5-HT neurons (irregular slow-firing and fast-firing) exhibited significant coherence and fired most during the active component of the SWA. These findings indicate that within the DRN electrophysiologically and neurochemically discrete neuronal groups exhibit distinct relations to cortical activity.


Duloxetine and Subacute Pain after Knee Arthroplasty when Added to a Multimodal Analgesic Regimen: A Randomized, Placebo-controlled, Triple-blinded Trial.
Duloxetine is effective for chronic musculoskeletal and neuropathic pain, but there are insufficient data to recommend the use of antidepressants for postoperative pain. The authors hypothesized that administration of duloxetine for 15 days would reduce pain with ambulation at 2 weeks after total knee arthroplasty. In this triple-blinded, randomized, placebo-controlled trial, patients received either duloxetine or placebo for 15 days, starting from the day of surgery. Patients also received a comprehensive multimodal analgesic regimen including neuraxial anesthesia, epidural analgesia, an adductor canal block, meloxicam, and oxycodone/acetaminophen as needed. The primary outcome was the pain score (0 to 10 numeric rating scale) with ambulation on postoperative day 14. One hundred six patients were randomized and analyzed. On day 14, duloxetine had no effect on pain with ambulation; mean pain was 3.8 (SD, 2.3) for placebo versus 3.5 (SD, 2.1) for duloxetine (difference in means [95% CI], 0.4 [-0.5 to 1.2]; P = 0.386). Symptoms potentially attributable to duloxetine discontinuation at study drug completion (nausea, anxiety) occurred among nine patients (duloxetine) and five patients (placebo); this was not statistically significant (P = 0.247). Statistically significant secondary outcomes included opioid consumption (difference in mean milligram oral morphine equivalents [95% CI], 8.7 [3.3 to 14.1], P = 0.002 by generalized estimating equation) over the postoperative period and nausea on day 1 (P = 0.040). There was no difference in other side effects or in anxiety and depression scores. When included as a part of a multimodal analgesic regimen for knee arthroplasty, duloxetine does not reduce subacute pain with ambulation.


Manipulation of skin temperature improves nocturnal sleep in narcolepsy.
Besides excessive daytime sleepiness, disturbed nocturnal sleep is a major complaint of patients with narcolepsy. Previously, alterations in skin temperature regulation in narcoleptic patients have been shown to be related to increased sleepiness. This study tests the hypothesis that direct control of nocturnal skin temperature might be applied to improve the disturbed sleep of narcoleptic patients. Participants were eight patients (five males) diagnosed as having narcolepsy with cataplexy according to the ICSD-2 criteria, mean (SD) age 28.6 (6.4) years, range 18-35 years. During two nights, sleep was recorded polysomnographically while proximal and distal skin temperature were manipulated using a comfortable thermosuit that induced skin temperature to cycle slowly with an amplitude of only 0.4 degrees C within the comfortable range normally observed during sleep. Logistic regression was used to evaluate the effect of skin temperature manipulation on the probability of occurrence of different sleep stages and nocturnal wakefulness. Proximal skin warming significantly suppressed wakefulness and enhanced slow wave sleep (SWS). In contrast, distal skin warming enhanced wakefulness and stage 1 sleep at the cost of SWS and REM sleep. The optimal combination of proximal skin warming and distal skin cooling led to a 160% increase in SWS, a 50% increase in REM sleep and a 68% decrease in wakefulness, compared with the least beneficial combination of proximal skin cooling and distal skin warming. Subtle skin temperature manipulations under controlled conditions significantly improved the typical nocturnal sleep problems in narcolepsy.


Planar microcoil-based microfluidic NMR probes.
Microfabricated small-volume NMR probes consisting of electroplated planar microcoils integrated on a glass substrate with etched microfluidic channels are fabricated and tested. 1H NMR spectra are acquired at 300 MHz with three different probes having observed sample volumes of respectively 30, 120, and 470 nL. The achieved sensitivity enables acquisition of an 1H spectrum of 160 microg sucrose in D2O, corresponding to a proof-of-concept for on-chip NMR spectroscopy. Increase of mass-sensitivity with coil diameter reduction is demonstrated experimentally for planar microcoils. Models that enable quantitative prediction of the signal-to-noise ratio and of the influence of microfluidic channel geometry on spectral resolution are presented and successfully compared to the experimental data. The main factor presently limiting sensitivity for high-resolution applications is identified as being probe-induced static magnetic field distortions. Finally, based on the presented model and measured data, future performance of planar microcoil-based microfluidic NMR probes is extrapolated and discussed.


Measurement of large spiral and target waves in chemical reaction-diffusion-advection systems: turbulent diffusion enhances pattern formation.
In the absence of advection, reaction-diffusion systems are able to organize into spatiotemporal patterns, in particular spiral and target waves. Whenever advection is present that can be parametrized in terms of effective or turbulent diffusion D(*), these patterns should be attainable on a much greater, boosted length scale. However, so far, experimental evidence of these boosted patterns in a turbulent flow was lacking. Here, we report the first experimental observation of boosted target and spiral patterns in an excitable chemical reaction in a quasi-two-dimensional turbulent flow. The wave patterns observed are ~50 times larger than in the case of molecular diffusion only. We vary the turbulent diffusion coefficient D(*) of the flow and find that the fundamental Fisher-Kolmogorov-Petrovsky-Piskunov equation, v(f) proportional sqrt[D(*)], for the asymptotic speed of a reactive wave remains valid. However, not all measures of the boosted wave scale with D(*) as expected from molecular diffusion, since the wave fronts turn out to be highly filamentous.


Prognostic value of high-sensitivity cardiac troponin I in heart failure patients with mid-range and reduced ejection fraction.
The identification of high-risk heart failure (HF) patients makes it possible to intensify their treatment. Our aim was to determine the prognostic value of a newly developed, high-sensitivity troponin I assay (Atellica®, Siemens Healthcare Diagnostics) for patients with HF with reduced ejection fraction (HFrEF; LVEF < 40%) and HF with mid-range EF (HFmrEF) (LVEF 40%-49%). A total of 520 patients with HFrEF and HFmrEF were enrolled in this study. Two-year all-cause mortality, heart transplantation, and/or left ventricular assist device implantation were defined as the primary endpoints (EP). A logistic regression analysis was used for the identification of predictors and development of multivariable models. The EP occurred in 14% of the patients, and these patients had higher NT-proBNP (1,950 vs. 518 ng/l; p < 0.001) and hs-cTnI (34 vs. 17 ng/l, p < 0.001) levels. C-statistics demonstrated that the optimal cut-off value for the hs-cTnI level was 17 ng/l (AUC 0.658, p < 0.001). Described by the AUC, the discriminatory power of the multivariable model (NYHA > II, NT-proBNP, hs-cTnI and urea) was 0.823 (p < 0.001). Including heart failure hospitalization as the component of the combined secondary endpoint leads to a diminished predictive power of increased hs-cTnI. hs-cTnI levels ≥ 17 ng/l represent an independent increased risk of an adverse prognosis for patients with HFrEF and HFmrEF. Determining a patient's hs-cTnI level adds prognostic value to NT-proBNP and clinical parameters.


Nanomedicine for respiratory diseases.
Treatment of respiratory diseases and infections has proved to be a challenging task, with the incidence of these ailments increasing worldwide. Nanotechnology-based drug and gene delivery systems offer a possible solution to some of the shortfalls of the current treatment regimen. Nanobased drug delivery systems have revolutionised the field of pharmacotherapy by presenting the ability to alter the pharmacokinetics of the conventional drugs to extend the drug retention time, reduce the toxicity and increase the half-life of the drugs. Delivery of exogenous genes to the airway epithelium in vivo has been limited by several physiological barriers, resulting in the low success rate of these systems. With the advent of nanotechnology, DNA compacted with cationic polymers to produce nanoparticles has exhibited a significant increase in the transfection efficiencies. With nanoparticulate drug/gene delivery systems, specific cells can be targeted by functionalising the polymeric nanoparticles with ligands that allow the particles to dock at a specific site of the cell. In addition, polymeric systems allow for the cargo to be released in a controlled and stimuli-responsive manner. The advantages that nanoparticulate delivery systems present in the treatment of respiratory diseases and infections are summarised in this review.


Branched dimerization of Tat peptide improves permeability to HeLa and hippocampal neuronal cells.
A dimeric branched peptide TATp-D designed as an analogue of the HIV-Tat protein transduction domain (TATp), a prototypical cell penetrating peptide (CPP), demonstrates significantly enhanced cell uptake at 0.25 to 2.5 μM. Live cell confocal laser scanning microscopy revealed that multivalency dramatically improved the permeation potency of TATp-D to HeLa and primary hippocampal neuronal cells. The observed enhanced ability of TATp-D to translocate through the membrane is highlighted by a non-linear dependence on concentration, exhibiting the greatest uptake at sub-micromolar concentrations as compared to TATp. Multimerization via bis-Fmoc Lysine offered a synthetically straightforward method to investigate the effects of multivalent CPPs while offering orthogonal handles for cargo attachment, increasing the utility of CPPs at significantly lower concentrations.


Mapping support policies for informal carers across the European Union.
At a time when health and social care services in European countries are under pressure to contain or cut costs, informal carers are relied upon as the main providers of long-term care. However, still little is known about the availability of direct and indirect support for informal carers across the European Union. Primary data collection in all EU member states was supplemented with an extensive review of the available literature. Various forms and levels of support have been implemented across Europe to facilitate the role of informal caregivers. Financial support is the most common type of support provided, followed by respite care and training. Most countries do not have a process in place to systematically identify informal carers and to assess their needs. Policies are often at an early stage of development and the breadth of support varies significantly across the EU. Policy developments are uneven across the member states, with some countries having mechanisms in place to assess the needs and support informal carers while others are only starting to take an interest in developing support services. Given the unprecedented challenges posed by population ageing, further research and better data are needed to capture and monitor information on informal carers, to help design adequate support policies and eventually to evaluate their impact across the EU.


Cannabinoid Ligands and Alcohol Addiction: A Promising Therapeutic Tool or a Humbug?
The vast therapeutic potential of cannabinoids of both synthetic and plant-derived origins currently makes these compounds the focus of a growing interest. Although cannabinoids are still illicit drugs, their possible clinical usefulness, including treatment of acute or neuropathic pain, have been suggested by several studies. In addition, some observations indicate that cannabinoid receptor antagonists may be useful for the treatment of alcohol dependence and addiction, which is a major health concern worldwide. While the synergism between alcohol and cannabinoid agonists (in various forms) creates undesirable side effects when the two are consumed together, the administration of CB1 antagonists leads to a significant reduction in alcohol consumption. Furthermore, cannabinoid antagonists also mitigate alcohol withdrawal symptoms. Herein, we present an overview of studies focusing on the effects of cannabinoid ligands (agonists and antagonists) during acute or chronic consumption of ethanol.


Tumor mutation burden forecasts outcome in ovarian cancer with BRCA1 or BRCA2 mutations.
Increased number of single nucleotide substitutions is seen in breast and ovarian cancer genomes carrying disease-associated mutations in BRCA1 or BRCA2. The significance of these genome-wide mutations is unknown. We hypothesize genome-wide mutation burden mirrors deficiencies in DNA repair and is associated with treatment outcome in ovarian cancer. The total number of synonymous and non-synonymous exome mutations (Nmut), and the presence of germline or somatic mutation in BRCA1 or BRCA2 (mBRCA) were extracted from whole-exome sequences of high-grade serous ovarian cancers from The Cancer Genome Atlas (TCGA). Cox regression and Kaplan-Meier methods were used to correlate Nmut with chemotherapy response and outcome. Higher Nmut correlated with a better response to chemotherapy after surgery. In patients with mBRCA-associated cancer, low Nmut was associated with shorter progression-free survival (PFS) and overall survival (OS), independent of other prognostic factors in multivariate analysis. Patients with mBRCA-associated cancers and a high Nmut had remarkably favorable PFS and OS. The association with survival was similar in cancers with either BRCA1 or BRCA2 mutations. In cancers with wild-type BRCA, tumor Nmut was associated with treatment response in patients with no residual disease after surgery. Tumor Nmut was associated with treatment response and with both PFS and OS in patients with high-grade serous ovarian cancer carrying BRCA1 or BRCA2 mutations. In the TCGA cohort, low Nmut predicted resistance to chemotherapy, and for shorter PFS and OS, while high Nmut forecasts a remarkably favorable outcome in mBRCA-associated ovarian cancer. Our observations suggest that the total mutation burden coupled with BRCA1 or BRCA2 mutations in ovarian cancer is a genomic marker of prognosis and predictor of treatment response. This marker may reflect the degree of deficiency in BRCA-mediated pathways, or the extent of compensation for the deficiency by alternative mechanisms.


Detection of platelet-monocyte aggregates by the ADAM(®) image cytometer.
Inappropriate platelet activation is known to be associated with various thrombotic disorders. Platelet-monocyte aggregates (PMAs), whose formation is mediated by platelet surface P-selectin (CD62P), can be used as a reliable marker to detect platelet activation. Previous studies have generally detected PMAs through flow cytometry-based approaches. Recently, the ADAM(®) image cytometer (Nanoentek Inc., Seoul, Korea) was developed for image-based cellular analysis. In this study, we detected PMAs with the ADAM(®) cytometer, evaluated the reproducibility of the measurements made by the ADAM(®) cytometer, and compared the abilities of the ADAM(®) cytometer and a flow cytometric assay to detect PMAs. Whole blood samples were collected from patients. Within 5 minutes of collection, anticoagulated whole blood samples were fixed in 10% paraformaldehyde and 5% glyoxal. Nineteen clinical specimens were collected; each was analyzed three times with the ADAM(®) cytometer in order to assess the reproducibility of its measurements. To compare the ability of the ADAM(®) cytometer with that of a flow cytometer to detect PMAs, each cytometer was used for 23 clinical samples and the correlation of the measurements was determined. The PMA measurements made by the ADAM(®) cytometer showed good reproducibility (CV < 10% for all specimens). Moreover, the PMA measurements made by the ADAM(®) cytometer exhibited a high correlation with those made by a flow cytometric assay (R = 0.944). The ADAM(®) cytometer is a suitable alternative method to the flow cytometry-based assays. Since the ADAM cytometer does not need specialized instrument knowledge or software proficiency (unlike flow cytometry), the ADAM(®) cytometer can be used as a rapid and reliable POCT device to measure platelet activation in peripheral blood. This, in turn, will provide valuable information regarding patient propensities to thrombotic diseases.


A longitudinal analysis of women's salivary testosterone and intrasexual competitiveness.
Research on within-subject changes in women's intrasexual competitiveness has generally focused on possible relationships between women's intrasexual competitiveness and estimates of their fertility. While this approach is useful for testing hypotheses about the adaptive function of changes in women's intrasexual competitiveness, it offers little insight into the proximate mechanisms through which such changes might occur. To investigate this issue, we carried out a longitudinal study of the hormonal correlates of changes in intrasexual competitiveness in a large sample of heterosexual women (N=136). Each woman provided saliva samples and completed an intrasexual competitiveness questionnaire in five weekly test sessions. Multilevel modeling of these data revealed a significant, positive within-subject effect of testosterone on intrasexual competitiveness, indicating that women reported greater intrasexual competitiveness when testosterone was high. By contrast, there were no significant effects of estradiol, progesterone, estradiol-to-progesterone ratio, or cortisol and no significant effects of any hormones on reported relationship jealousy. This is the first study to demonstrate correlated changes in measured testosterone levels and women's reported intrasexual competitiveness, implicating testosterone in the regulation of women's intrasexual competitiveness.


Improvement in cochlear flow with Pycnogenol® in patients with tinnitus: a pilot evaluation.
The aim of this preliminary evaluation was to study the efficacy of Pycnogenol in improving cochlear flow in patients with mild-to-moderate tinnitus present for at least two weeks (without vertigo or important hearing loss), possibly associated with cochlear hypo-perfusion. Patients with mild-to-moderate, idiopatic, monolateral tinnitus present for at least 2 weeks were included; no vertigo or important hearing loss had been found in a specific examination. The origin of tinnitus had been sudden (hours or days). Fifty-eight patients used Pycnogenol: 24 used 150 mg/day (group A; mean age 43.2+/-4.3) and 34 patients 100 mg/day (group B: mean age 42.4+/-3.8). Controls included 24 patients (mean age 42.3+/-4.5). The groups were comparable for their clinical problem and age and sex. The average duration of treatment was 34.3+/-3.1 days. No side effects were observed and no drop-outs occurred. The variations in cochlear flow velocity (in cm/s at the cochlear artery), at inclusion and after four weeks of treatment indicated that flow velocity at the level of the affected ear was significantly lower (both the diastolic and systolic components; P<0.05) in comparison with the other ear. The treatment favored an improvement in systolic (P<0.05) and diastolic flow velocity (P<0.05) in the two treatment groups A+B. The increase in flow velocity was very limited and not significant in controls. These results suggest that in selected patients with tinnitus and altered perfusion, Pycnogenol is effective in a short period of time in relieving tinnitus symptoms by improving cochlear blood flow. The effect is more pronounced with higher Pycnogenol dosage. More studies should be planned to better evaluate the pathology and potential applications of Pycnogenol in a larger number of patients who are currently without a real therapeutic solution.


Effects of acute stressors on the expression of oxytocin receptor mRNA in hearts of rats with different activity of HPA axis.
Cardiovascular system is regulated by a diverse array of hormones, neurotransmitters and neuropeptides. Oxytocin and its receptors (OTR) were also shown to regulate cardiovascular functions and this hormone was even called cardiovascular hormone. In recent publication, we demonstrated the expression of mRNA of OTR by real-time quantitative PCR (RT qPCR) in all rat heart compartments. The aim of this study was to investigate the effects of acute restraint stress on OTR mRNA expression in two rat strains with different activity of HPA axis. Adult male Sprague-Dawley and Lewis rats, the latter strain reported to have lower HPA activity, were used in RT qPCR studies and Wistar rats in immunofluorescent ones. Both acute restraint (IS) and this stress combined with the immersion of rats in water (ICS) lasted 60 min. Gene expression of OTR mRNA was estimated in all heart compartments after 1 or 3 hours after stress termination (IS1, IS3, ICS1, ICS3). The relative expression was calculated using 2(-ΔΔC)T method. In immunofluorescent studies we used commercial specific OTR antibodies. In RT qPCR studies we found higher expression of OTR mRNA in atria than in ventricles and no statistical differences between Sprague-Dawley and Lewis rats under basal conditions. Relative expression of OTR mRNA after 60 min lasting stress exposure differed in dependence on the stress type and partly on the time interval after the stress termination. When compared to controls, in rat left atria both stressors caused inhibition of OTR mRNA expression in both rat strains. In rat ventricles, which have very low OTR mRNA expression, there was a significant difference in the effect of two stressors. In most groups ICS displayed the increase of OTR mRNA expression if compared to IS groups. Immunofluorescent studies revealed changes induced by acute restraint stress in all heart compartments. The immunofluorescent studies suggested that acute stress induces higher colocalization of OTR with the nuclei than it was observed in the controls. The expression of OTR mRNA in all heart compartments of controls as well as after stress exposure in Sprague-Dawley and Lewis rats support the notion that OTR plays a regulatory role in the cardiovascular system and is also involved in the regulations in the heart after stress. The immunofluorescent observation that OTRs coexpress in areas of cell nuclei in certain heart compartments and after acute stress, compared to controls, requires further studies.


Human immunodeficiency virus Type 1 nucleocapsid protein (NCp7) directs specific initiation of minus-strand DNA synthesis primed by human tRNA(Lys3) in vitro: studies of viral RNA molecules mutated in regions that flank the primer binding site.
Retroviral reverse transcription starts near the 5' end of unspliced viral RNA at a sequence called the primer binding site (PBS), where the tRNA primer anneals to the RNA template for initiation of DNA synthesis. We have investigated the roles of NCp7 in annealing of primer tRNA(Lys3) to the PBS and in reverse transcriptase (RT) activity, using a cell-free reverse transcription reaction mixture consisting of various 5' viral RNA templates, natural primer tRNA(Lys3) or synthetic primer, human immunodeficiency virus type I (HIV-1) nucleocapsid protein (NCp7), and HIV-1 RT. In the presence of tRNA(Lys3), NCp7 was found to stimulate synthesis of minus-strand strong-stop DNA [(-)ssDNA], consistent with previous reports. However, specific DNA synthesis was observed only at a NCp7/RNA ratio similar to that predicted to be present in virions. Moreover, at these concentrations, NCp7 inhibited the synthesis of nonspecific reverse-transcribed DNA products, which are initiated because of self-priming by RNA templates. In contrast to results obtained with tRNA(Lys3) as primer, NCp7 inhibited the synthesis of (-)ssDNA products primed by an 18-nucleotide (nt) ribonucleotide (rPR), complementary to the PBS, even though rPR can initiate synthesis of such material in the absence of preannealing with NCp7. Primer placement band shift assays showed that NCp7 was necessary for efficient formation of the tRNA-RNA complex. In contrast, NCp7 was found to prevent formation of the rPR-RNA complex. Since NCp7 appears to exert opposite effects (annealing versus dissociation) on tRNA(Lys3) and rPR substrates, the non-PBS binding regions of the tRNA(Lys3) molecule may play a role in the annealing of tRNA to the template. We also investigated the roles of an A-rich loop upstream of the PBS, a 7-nt region immediately downstream of the PBS, and a 54-nt deletion further downstream of the PBS in interactions with tRNA(Lys3). We found that deletions in the 54-nt region that may prevent formation of the U5-leader stem prevented tRNA(Lys3) placement and priming, while deletions in the A-rich loop or the 7-nt sequence had relatively minor effects in this regard.


Guide to red fluorescent proteins and biosensors for flow cytometry.
Since the discovery of the first red fluorescent protein (RFP), named DsRed, 12 years ago, a wide pallet of red-shifted fluorescent proteins has been cloned and biotechnologically developed into monomeric fluorescent probes for optical microscopy. Several new types of monomeric RFPs that change the emission wavelength either with time, called fluorescent timers, or after a brief irradiation with violet light, known as photoactivatable proteins, have been also engineered. Moreover, RFPs with a large Stokes shift of fluorescence emission have been recently designed. Because of their distinctive excitation and fluorescence detection conditions developed specifically for microscopy, these fluorescent probes can be suboptimal for flow cytometry. Here, we have selected and summarized the advanced orange, red, and far-red fluorescent proteins with the properties specifically required for the flow cytometry applications. Their effective brightness was calculated for the laser sources available for the commercial flow cytometers and sorters. Compatibility of the fluorescent proteins of different colors in a multiparameter flow cytometry was determined. Novel FRET pairs, utilizing RFPs, RFP-based intracellular biosensors, and their application to a high-throughput screening, are also discussed.


MAST: a flexible statistical framework for assessing transcriptional changes and characterizing heterogeneity in single-cell RNA sequencing data.
Single-cell transcriptomics reveals gene expression heterogeneity but suffers from stochastic dropout and characteristic bimodal expression distributions in which expression is either strongly non-zero or non-detectable. We propose a two-part, generalized linear model for such bimodal data that parameterizes both of these features. We argue that the cellular detection rate, the fraction of genes expressed in a cell, should be adjusted for as a source of nuisance variation. Our model provides gene set enrichment analysis tailored to single-cell data. It provides insights into how networks of co-expressed genes evolve across an experimental treatment. MAST is available at https://github.com/RGLab/MAST .


Safety and efficacy of transradial coronary angiography and intervention in patients older than 80 years: from the Korean Transradial Intervention Prospective Registry.
Radial artery access for coronary procedures is a safe and beneficial technique. However, elderly patients have been considered as a higher risk group of access site related complications compared to younger patients. This study was conducted to investigate the feasibility and safety of transradial coronary angiography or intervention in the elderly. A total of 6132 patients from Korean Transradial Intervention Prospective Registry at 20 centers were analyzed. Patients were divided into the non-elderly group (n = 5667) and the elderly (≥ 80 years) group (n = 465). Using propensity score matching, the elderly group (n = 465) was compared with one-to-one matched the non-elderly group (n = 465). After propensity score matching, mean age was 64.3 ± 10.3 years in the non-elderly group and 83.5 ± 3.3 years in the elderly group. There was no difference of procedural characteristics, procedural and fluoroscopic times. Access site cross-over rate was not different between the non-elderly group and elderly group (7.5% vs. 6.2%, P = 0.074). Bleeding complications occurred similarly in two groups (2.6% of the non-elderly group vs. 1.9% of the elderly group, P = 0.660). Access site complications were 1.9% of the non-elderly group and 0.9% of the elderly group (P = 0.263). Both of in hospital death and cardiovascular death for one year were also similar between two groups. Transradial angiography or intervention was safe and feasible in elderly patients. Complication rates and clinical outcomes in elderly patients were comparable with those in non-elderly patients.


Navicular drop as a composite measure of excessive pronation.
The measure of navicular drop has been used as an indicator of pronation at the foot. It is defined as the distance the navicular tuberosity moves in standing, as the subtalar joint is allowed to move from its neutral position to a relaxed position. The purposes of this study were to test the reliability of a method to measure navicular drop and to assess the relationships among measures of forefoot to rearfoot position, subtalar joint neutral position, and navicular drop. The results support traditional biomechanical theory but indicate that other factors contribute significantly to navicular drop.


The glucagon-like peptide 2 receptor is expressed in enteric neurons and not in the epithelium of the intestine.
Glucagon-like peptide 2 (GLP-2) is a potent intestinotrophic growth factor with therapeutic potential in the treatment of intestinal deficiencies. It has recently been approved for the treatment of short bowel syndrome. The effects of GLP-2 are mediated by specific binding of the hormone to the GLP-2 receptor (GLP-2R) which was cloned in 1999. However, consensus about the exact receptor localization in the intestine has never been established. By physical, chemical and enzymatic tissue fragmentation, we were able to divide rat jejunum into different compartments consisting of: (1) epithelium alone, (2) mucosa with lamina propria and epithelium, (3) the external muscle coat including myenteric plexus, (4) a compartment enriched for the myenteric plexus and (5) intestine without epithelium. Expression of Glp2r; chromogranin A; tubulin, beta 3; actin, gamma 2, smooth muscle, enteric and glial fibrillary acidic protein in these isolated tissue fractions was quantified with qRT-PCR. Expression of the Glp2r was confined to compartments containing enteric neurons and receptor expression was absent in the epithelium. Our findings provide evidence for the expression of the GLP-2R in intestinal compartments rich in enteric neurons and, importantly they exclude significant expression in the epithelium of rat jejunal mucosa.


Prediction of protein solvent profile using SVR.
We describe a support vector regression (SVR) approach to predict the accessible surface area (ASA) of a protein from its sequence. Our approach encodes each protein residue as a vector of amino acid propensities derived from a multiple alignment of the subject protein with homologous proteins. The vector consists of the log-likelihood ratios of each of the twenty amino acids in the residue's multiple alignment column. Using a reference set of proteins of known structure and, hence, known ASA, we trained an SVR model. Each training sample consists of the fifteen log-likelihood vectors in a window of width fifteen surrounding a residue, along with the "true" ASA value, computed from the known structure. To apply the model to proteins of unknown structure, only the subject protein sequence is required. Our method uses PSI-BLAST to simultaneously determine a set of (putative) homologs and compute the log-likelihood vectors needed to encode the subject protein. We show that this method provides substantially improved accuracy in predicting ASA when compared with an earlier method.


Insulin resistance is an independent correlate of increased urine albumin excretion: a cross-sectional study in Iranian Type 2 diabetic patients.
To assess the association of insulin resistance with increased urinary albumin excretion (UAE) in a cohort of Iranian Type 2 diabetic patients. Three hundred and sixty-one men and 472 women with Type 2 diabetes were enrolled from three different outpatient clinics (Tehran, Iran) during the period 2005-2008. Patients with obstructive uropathy, severe heart failure, liver disease, cancer, autoimmune disease and macroalbuminuria were not included. Microalbuminuria (MA; defined as UAE >or= 30 mg/day) was found in 242 (29.1%) patients; 591 (70.9%) subjects had normoalbuminuria (UAE < 30 mg/day). Insulin resistance was assessed using homeostasis model assessment of insulin resistance (HOMA-IR). HOMA-IR index values were higher in subjects with MA than those with normoalbuminuria (P < 0.00001). Adjusted values (for age, sex and duration of diabetes) of UAE and HOMA-IR were 11.81 +/- 7.51 (mg/day) and 3.30 +/- 2.21 in normoalbuminuric and 75.36 +/- 55.57 (mg/day) and 4.98 +/- 3.22 in the MA group, respectively (P < 0.00001 for all). Multiple regression analysis showed that UAE was predicted by HOMA-IR, independently of age, duration of diagnosed diabetes, triglycerides, waist circumference, metabolic control, blood pressure and related treatments (P < 0.00001). When patients were categorized into quartiles of HOMA-IR, those of the fourth quartile (i.e. the most insulin resistant) were at a higher risk of increased UAE than other quartiles [odds ratio (OR) 3.7 (95% confidence intervals 2.7-6.2)]. In Iranian Type 2 diabetic patients, albuminuria was strongly associated with insulin resistance. HOMA-IR is an independent predictor of UAE.


Cholinergic nucleus basalis tauopathy emerges early in the aging-MCI-AD continuum.
The cholinergic denervation in Alzheimer's disease (AD) provides the rationale for treatments with anticholinesterases. The presence of this cholinergic lesion is solidly established in advanced AD. Whether it also exists in early disease remains unsettled. This question was addressed with thioflavin-S histofluorescence to identify neurofibrillary tangles (NFT) and two tau antibodies (AT8, Alz-50) to identify pre-tangle cytopathology in the nucleus basalis, the source of cortical cholinergic innervation. Methods for the concurrent visualization of tauopathy and choline acetyltransferase were used to determine if the cytopathology was selectively located within cholinergic neurons. Five elderly index cases who had died at the stage of mild cognitive impairment (MCI) or early AD were identified by longitudinal neuropsychological and behavioral assessments. They were compared to 7 age-matched cognitively normal subjects. NFT and AT8 (or Alz-50) immunostaining in cholinergic nucleus basalis neurons existed even in the cognitively normal subjects. The percentage of tauopathy-containing nucleus basalis neurons was greater in the cognitively impaired and showed a significant correlation with memory scores obtained 1-18 months prior to death. These results show that cytopathology in cortical cholinergic pathways is a very early event in the course of the continuum that leads from advanced age to MCI and AD.


Neratinib after trastuzumab-based adjuvant therapy in patients with HER2-positive breast cancer (ExteNET): a multicentre, randomised, double-blind, placebo-controlled, phase 3 trial.
Neratinib, an irreversible tyrosine-kinase inhibitor of HER1, HER2, and HER4, has clinical activity in patients with HER2-positive metastatic breast cancer. We aimed to investigate the efficacy and safety of 12 months of neratinib after trastuzumab-based adjuvant therapy in patients with early-stage HER2-positive breast cancer. We did this multicentre, randomised, double-blind, placebo-controlled, phase 3 trial at 495 centres in Europe, Asia, Australia, New Zealand, and North and South America. Eligible women (aged ≥18 years, or ≥20 years in Japan) had stage 1-3 HER2-positive breast cancer and had completed neoadjuvant and adjuvant trastuzumab therapy up to 2 years before randomisation. Inclusion criteria were amended on Feb 25, 2010, to include patients with stage 2-3 HER2-positive breast cancer who had completed trastuzumab therapy up to 1 year previously. Patients were randomly assigned (1:1) to receive oral neratinib 240 mg per day or matching placebo. The randomisation sequence was generated with permuted blocks stratified by hormone receptor status (hormone receptor-positive [oestrogen or progesterone receptor-positive or both] vs hormone receptor-negative [oestrogen and progesterone receptor-negative]), nodal status (0, 1-3, or ≥4), and trastuzumab adjuvant regimen (sequentially vs concurrently with chemotherapy), then implemented centrally via an interactive voice and web-response system. Patients, investigators, and trial sponsors were masked to treatment allocation. The primary outcome was invasive disease-free survival, as defined in the original protocol, at 2 years after randomisation. Analysis was by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00878709. Between July 9, 2009, and Oct 24, 2011, we randomly assigned 2840 women to receive neratinib (n=1420) or placebo (n=1420). Median follow-up time was 24 months (IQR 20-25) in the neratinib group and 24 months (22-25) in the placebo group. At 2 year follow-up, 70 invasive disease-free survival events had occurred in patients in the neratinib group versus 109 events in those in the placebo group (stratified hazard ratio 0·67, 95% CI 0·50-0·91; p=0·0091). The 2-year invasive disease-free survival rate was 93·9% (95% CI 92·4-95·2) in the neratinib group and 91·6% (90·0-93·0) in the placebo group. The most common grade 3-4 adverse events in patients in the neratinib group were diarrhoea (grade 3, n=561 [40%] and grade 4, n=1 [<1%] vs grade 3, n=23 [2%] in the placebo group), vomiting (grade 3, n=47 [3%] vs n=5 [<1%]), and nausea (grade 3, n=26 [2%] vs n=2 [<1%]). QT prolongation occurred in 49 (3%) patients given neratinib and 93 (7%) patients given placebo, and decreases in left ventricular ejection fraction (≥grade 2) in 19 (1%) and 15 (1%) patients, respectively. We recorded serious adverse events in 103 (7%) patients in the neratinib group and 85 (6%) patients in the placebo group. Seven (<1%) deaths (four patients in the neratinib group and three patients in the placebo group) unrelated to disease progression occurred after study drug discontinuation. The causes of death in the neratinib group were unknown (n=2), a second primary brain tumour (n=1), and acute myeloid leukaemia (n=1), and in the placebo group were a brain haemorrhage (n=1), myocardial infarction (n=1), and gastric cancer (n=1). None of the deaths were attributed to study treatment in either group. Neratinib for 12 months significantly improved 2-year invasive disease-free survival when given after chemotherapy and trastuzumab-based adjuvant therapy to women with HER2-positive breast cancer. Longer follow-up is needed to ensure that the improvement in breast cancer outcome is maintained. Wyeth, Pfizer, Puma Biotechnology.


Chaoborus and gasterosteus anti-predator responses in Daphnia pulex are mediated by independent cholinergic and gabaergic neuronal signals.
Many prey species evolved inducible defense strategies that protect effectively against predation threats. Especially the crustacean Daphnia emerged as a model system for studying the ecology and evolution of inducible defenses. Daphnia pulex e.g. shows different phenotypic adaptations against vertebrate and invertebrate predators. In response to the invertebrate phantom midge larvae Chaoborus (Diptera) D. pulex develops defensive morphological defenses (neckteeth). Cues originating from predatory fish result in life history changes in which resources are allocated from somatic growth to reproduction. While there are hints that responses against Chaoborus cues are transmitted involving cholinergic neuronal pathways, nothing is known about the neurophysiology underlying the transmission of fish related cues. We investigated the neurophysiological basis underlying the activation of inducible defenses in D. pulex using induction assays with the invertebrate predator Chaoborus and the three-spined stickleback Gasterosteus aculeatus. Predator-specific cues were combined with neuro-effective substances that stimulated or inhibited the cholinergic and gabaergic nervous system. We show that cholinergic-dependent pathways are involved in the perception and transmission of Chaoborus cues, while GABA was not involved. Thus, the cholinergic nervous system independently mediates the development of morphological defenses in response to Chaoborus cues. In contrast, only the inhibitory effect of GABA significantly influence fish-induced life history changes, while the application of cholinergic stimulants had no effect in combination with fish related cues. Our results show that cholinergic stimulation mediates signal transmission of Chaoborus cues leading to morphological defenses. Fish cues, which are responsible for predator-specific life history adaptations involve gabaergic control. Our study shows that both pathways are independent and thus potentially allow for adjustment of responses to variable predation regimes.


Age-Related Declines in the Ability to Modulate Common Input to Bilateral and Unilateral Plantar Flexors During Forward Postural Lean.
Aging can impair an ability to lean the body forward to the edge of the base of support. Here, we investigated, using a coherence analysis, common inputs to bilateral and unilateral plantar flexor muscles to test a hypothesis that the age-related impairment would be related to strong synchronous bilateral activation and reduced cortical control of these muscles. Healthy young (n = 14) and elderly adults (n = 19), who were all right-foot dominant, performed quiet standing task and tasks that required the subjects to lean their body forward to 35 and 75% of the maximum lean distance. The electromyogram was recorded from the bilateral medial gastrocnemius (MG) and soleus (SL) muscles. We analyzed delta-band coherence, that reflects comodulation of muscle activity, between the bilateral homologous muscles (MG-MG and SL-SL pairs). The origin of this bilateral comodulation is suggested to be the subcortical system. Also, we examined beta-band coherence, that is related to the corticospinal drive, between the unilateral muscles (MG-SL pair) in the right leg. Results indicated that the bilateral delta-band coherence for the MG-MG pair was significantly smaller in the 75% forward lean than quiet standing and 35% forward lean tasks for the young adults (quiet: p = 0.036; 35%: p = 0.0011). The bilateral delta-band coherence for the SL-SL pair was significantly smaller in the 75% forward lean than 35% forward lean task for the young adults (p = 0.027). Furthermore, the unilateral beta-band coherence was larger in the forward lean than quiet standing task for the young adults (35%: p < 0.001; 75%: p = 0.029). Contrarily, the elderly adults did not demonstrate such changes. These findings suggest the importance of decreasing the synchronous bilateral activation and increasing the unilateral cortical control of the plantar flexor muscles for the successful forward postural lean performance, and that aging impairs this modulatory ability.


Positive effect of flavonoids to cardiovascular and central nervous system.
Moderate intake of wine may be protective against various neurologic, cardiovascular and gastroenteric diseases. Low concentration of alcohol (wine diluted with water 1:2 to 1:10) exhibits strong antimicrobial activity against majority of gastrointestinal pathogens as well as against Chlamydia pneumoniae. In contrast, higher doses of even diluted alcohol (more than 5-7 glasses a day) may cause severe neuropsychic disorders with major social consequences. Wine contains variety of molecules with cardioprotective effect and antiinfectious properties.


Transgenerational effect of neighborhood poverty on low birth weight among African Americans in Cook County, Illinois.
In perinatal epidemiology, transgenerational risk factors are defined as conditions experienced by one generation that affect the pregnancy outcomes of the next generation. The authors investigated the transgenerational effect of neighborhood poverty on infant birth weight among African Americans. Stratified and multilevel logistic regression analyses were performed on an Illinois transgenerational data set with appended US Census income information. Singleton African-American infants (n = 40,648) born in 1989-1991 were considered index births. The mothers of index infants had been born in 1956-1976. The maternal grandmothers of index infants were identified. Rates of infant low birth weight (<2,500 g) rose as maternal grandmother's residential environment during her pregnancy deteriorated, independently of mother's residential environment during her pregnancy. In a multilevel logistic regression model that accounted for clustering by maternal grandmother's residential environment, the adjusted odds ratio (controlling for mother's age, education, prenatal care, cigarette smoking status, and residential environment) for infant low birth weight for maternal grandmother's residence in a poor neighborhood (compared with an affluent neighborhood) equaled 1.3 (95% confidence interval: 1.1, 1.4). This study suggests that maternal grandmother's exposure to neighborhood poverty during her pregnancy is a risk factor for infant low birth weight among African Americans.


Resource use patterns predict long-term outcomes of plant competition for nutrients and light.
An 11-year competition experiment among combinations of six prairie perennial plant species showed that resource competition theory generally predicted the long-term outcome of competition. We grew each species in replicated monocultures to determine its requirements for soil nitrate (R*) and light (I*). In six pairwise combinations, the species with the lower R* and I* excluded its competitor, as predicted by theory. In the remaining two pairwise combinations, one species had a lower R*, and the second had a lower I*; these species pairs coexisted, although it is unclear whether resource competition alone was responsible for their coexistence. Smaller differences in R* or I* between competing species led to slower rates of competitive exclusion, and the influence of R* differences on the rate of competitive exclusion was more pronounced on low-nitrogen soils, while the influence of I* differences was more pronounced on high-nitrogen (low-light) soils. These results were not explained by differences in initial species abundances or neutrality. However, only a few of our paired species coexisted under our experimentally imposed conditions (homogeneous soils, high seeding densities, minimal disturbance, regular water, and low herbivory levels), suggesting that other coexistence mechanisms help generate the diversity observed in natural communities.


Validation of the Atopic Dermatitis Control Tool (ADCT©) using a longitudinal survey of biologic-treated patients with atopic dermatitis.
The Atopic Dermatitis Control Tool (ADCT©) is a brief patient self-administered instrument designed and validated to assess atopic dermatitis (AD) control; six AD symptoms and impacts are evaluated over the past week, including overall severity of symptoms, days with intense episodes of itching, intensity of bother, problem with sleep, impact on daily activities, and impact on mood or emotions. This study assessed the reliability, validity, and responsiveness of the ADCT in a longitudinal context, and provided thresholds to identify meaningful within-person change. Data were from a prospective, longitudinal patient survey study of real-world effectiveness of dupilumab in patients with AD. Eligible patients completed a baseline survey before starting dupilumab and were followed at Months 1, 2, 3, and 6 post-initiation as they became eligible. Psychometric analyses confirmed internal consistency; Cronbach's α coefficients were consistently above the threshold of 0.70 across each follow-up; item-to-total correlations were above the threshold of r ≥ 0.50. High correlations between the ADCT and the Dermatology Life Quality Index (DLQI) and skin pain supported construct validity, while known-group validity was shown on Patient Global Assessment of Disease (PGAD) overall well-being subgroups with worse AD-related overall well-being having higher mean ADCT total scores at all time points. The ability of the ADCT to detect change was confirmed; the threshold for meaningful within-person change was estimated to be 5 points. Finally, test-retest reliability was confirmed in subgroups of patients with stable PGAD responses. Our findings confirm that the ADCT is a valid and reliable tool for assessing AD control.


A genetic analysis of group movement in an isolated population of tree-roosting bats.
Group fission is an important dispersal mechanism for philopatric adults. In Cypress Hills Interprovincial Park, Saskatchewan, tree-roosting big brown bats (Eptesicus fuscus) exhibit fission-fusion roosting behaviour. During 2004-2007, the majority of females previously resident to roosting area 1 (RA1) moved to a new roosting area (RA4). We examined how genetic relationships, inferred from data for microsatellite loci and mitochondrial DNA, influenced new roost area (RA) selection during 2006 when colony members were split between the RAs. We found that females who moved to RA4 had higher average relatedness than those that remained in RA1. We found that nearly all females belonging to matrilines with high average relatedness moved to RA4 while females from matrilines with low average relatedness were split between the two RAs. These results suggest that closely related maternal kin preferentially move to new RAs. However, daily roosting preferences within a RA are not based on genetic relationships probably because daily roosting associations between kin and non-kin are used to ensure adequate roost group size. Studying the effects of kinship on the fission and movements of groups not only enhances our understanding of social behaviour and population genetics but also informs conservation decisions.


Breast Cancer-Specific miR Signature Unique to Extracellular Vesicles Includes "microRNA-like" tRNA Fragments.
Extracellular vesicles (EV), including exosomes and shed vesicles, have been implicated in intercellular communication; however, their biomarker potential is less clear. Therefore, EVs derived from MCF7 and MCF10A cells were analyzed to identify unique miRNA (miR) profiles that distinguish their origin. One characteristic common to the miR profiles of MCF7 EVs and their parent cells is the high abundance of miR-21, let-7a, miR-100, and miR-125b, and low levels of miR-205. A second characteristic is the high abundance of "miRNA-like" tRNA fragments, which is unique to the MCF7 EVs, and is not found in comparing the cellular profiles. In addition, correlations were examined in the MCF7 cellular expression levels of these five miRs and two tRNA-derived miRNAs, miR-720 and miR-1274b, and compared with the correlations in MCF7 EV levels. Interestingly, correlations in the cellular expression of miR-125b, miR-100, and let-7a are mirrored in the EVs. In contrast, correlations in tRNA-derived miRNA levels are found only in the EVs. The findings suggest that EV miR clusters can be defined based on functional miR interactions related to correlated cellular expression levels or physical miR interactions, for example, aggregation due to comparable binding affinities to common targets. These results point to using high levels of tRNA-derived small RNA fragments in combination with known miR signatures of tumors to distinguish tumor-derived EVs in circulation from EVs derived from other cell sources. Such biomarkers would be unique to the EVs where high abundances of tRNA fragments are amplified with respect to their cellular levels.


Effect of solar ultraviolet radiation exposure on serum 25(OH)D concentration: a pilot randomised controlled trial.
Sunlight generates vitamin D, but there are scant human data from randomised trials on which to base health policy advice about how much sun exposure is necessary to change 25(OH)D concentrations. The purpose of the study was to evaluate the feasibility of using solar ultraviolet (UV) radiation exposure to generate a change in 25(OH)D concentration in a randomised controlled trial (RCT). The intervention tested in this RCT was supervised exposure to one standard erythemal dose (SED; 100 J m-2) of solar UV radiation three days per week for three weeks with approximately 35% of the body surface area not covered by clothing. Thirty-six fair-skinned (skin type II and III) indoor workers from Brisbane, Australia were randomised into either the intervention group (n = 16) or the control group (n = 20); the latter did not receive any supervised sun exposure. We asked both groups to use sunscreen and to minimise time outdoors during the study period. We collected blood samples at baseline, once per week during the three week intervention period, and four weeks after the intervention finished. The cumulative UV radiation exposure over the intervention period measured using polysulphone badges was higher in the intervention group than in the control group (median 8 vs. 4 SEDs, p = 0.14). After three weeks, the mean serum 25(OH)D concentration increased from 60 to 65 nmol l-1 in the intervention group and from 55 to 57 nmol l-1 in the control group. After adjustment for baseline 25(OH)D, the mean change per week during the intervention phase was non-significantly higher in the intervention than in the control group (0.7 vs. 0.3; p = 0.35). This difference was not sustained during the follow-up period. Large field trials are needed to inform policy about how much natural sun exposure is required to raise 25(OH)D concentrations. This pilot identified key issues that need to be considered in the design of such a trial.


Optical frequency synthesis from a cryogenic microwave sapphire oscillator.
We demonstrate an optical frequency comb with fractional frequency instability of </=2x10(-14) at measurement times near 1 s, when the 10th harmonic of the comb spacing is controlled by a liquid helium cooled microwave sapphire oscillator. The frequency instability of the comb is estimated by comparing it to a cavity-stabilized optical oscillator. The less conventional approach of synthesizing low-noise optical signals from a microwave source is relevant when a laboratory has microwave sources with frequency stability superior to their optical counterparts. We describe the influence of high frequency environmental noise and how it impacts the phase-stabilized frequency comb performance at integration times less than 1 s.


Ghost Ileostomy in Advanced Ovarian Cancer: A Reliable Option.
Diverting ileostomy (DI) has been proposed to reduce the incidence and consequences anastomotic leakage after bowel resection. In colorectal cancer treatment, ghost ileostomy (GI) has been proposed as an alternative to DI. Our objective was to report the results of GI associated with colorectal resection in the treatment of ovarian cancer. This is an observational pilot study performed in a single institution. The main objective sought was to report the results of GI associated with colorectal resection in the treatment of ovarian cancer: 26 patients were included. Modified posterior exenteration was performed in 24 cases (92.3%) and rectum resection in the 2 cases of relapse (7.7%). After the main procedure GI was created, to check up the anastomosis status, a sequential postoperative rectoscopy was performed on postoperative day 5 ± 1 (range, 4-7). Serum levels were monitored in first and third postoperative days just with a descriptive intention to establish its relationship with the rectoscopy findings. In 2 cases, rectoscopy demonstrated a leakage. During postoperative course, no other complication related with the GI or DI was observed. No case of clinical anastomotic leakage was found. To the extent of our knowledge, this is the first study in which GI has been performed for the treatment of patients with ovarian cancer. Ghost ileostomy represents a real option that may reduce the number of ileostomies performed in ovarian cancer without increasing the morbimortality. Ghost ileostomy presents the advantages of DI while avoiding its drawbacks. It also seems to be a safe, feasible, and reproducible technique that does not add significant costs to the surgery.


Ongoing cortical activity at rest: criticality, multistability, and ghost attractors.
The ongoing activity of the brain at rest, i.e., under no stimulation and in absence of any task, is astonishingly highly structured into spatiotemporal patterns. These spatiotemporal patterns, called resting state networks, display low-frequency characteristics (<0.1 Hz) observed typically in the BOLD-fMRI signal of human subjects. We aim here to understand the origins of resting state activity through modeling via a global spiking attractor network of the brain. This approach offers a realistic mechanistic model at the level of each single brain area based on spiking neurons and realistic AMPA, NMDA, and GABA synapses. Integrating the biologically realistic diffusion tensor imaging/diffusion spectrum imaging-based neuroanatomical connectivity into the brain model, the resultant emerging resting state functional connectivity of the brain network fits quantitatively best the experimentally observed functional connectivity in humans when the brain network operates at the edge of instability. Under these conditions, the slow fluctuating (<0.1 Hz) resting state networks emerge as structured noise fluctuations around a stable low firing activity equilibrium state in the presence of latent "ghost" multistable attractors. The multistable attractor landscape defines a functionally meaningful dynamic repertoire of the brain network that is inherently present in the neuroanatomical connectivity.


Neuroprotection Mediated through GluN2C-Containing N-methyl-D-aspartate (NMDA) Receptors Following Ischemia.
Post-ischemic activation of NMDA receptors (NMDARs) has been linked to NMDAR subunit-specific signaling that mediates pro-survival or pro-death activity. Although extensive studies have been performed to characterize the role of GluN2A and GluN2B following ischemia, there is less understanding regarding the regulation of GluN2C. Here, we show that GluN2C expression is increased in acute hippocampal slices in response to ischemia. Strikingly, GluN2C knockout mice, following global cerebral ischemia, exhibit greater neuronal death in the CA1 area of the hippocampus and reduced spatial working memory compared to wild-type mice. Moreover, we find that GluN2C-expressing hippocampal neurons show marked resistance to NMDA-induced toxicity and reduced calcium influx. Using both in vivo and in vitro experimental models of ischemia, we demonstrate a neuroprotective role of GluN2C, suggesting a mechanism by which GluN2C is upregulated to promote neuronal survival following ischemia. These results may provide insights into development of NMDAR subunit-specific therapeutic strategies to protect neurons from excitotoxicity.


Communications: Nanomagnetic shielding: High-resolution NMR in carbon allotropes.
The understanding and control of the magnetic properties of carbon-based materials is of fundamental relevance in applications in nano- and biosciences. Ring currents do play a basic role in those systems. In particular the inner cavities of nanotubes offer an ideal environment to investigate the magnetism of synthetic materials at the nanoscale. Here, by means of (13)C high resolution NMR of encapsulated molecules in peapod hybrid materials, we report the largest diamagnetic shifts (down to -68.3 ppm) ever observed in carbon allotropes, which is connected to the enhancement of the aromaticity of the nanotube envelope upon doping. This diamagnetic shift can be externally controlled by in situ modifications such as doping or electrostatic charging. Moreover, defects such as C-vacancies, pentagons, and chemical functionalization of the outer nanotube quench this diamagnetic effect and restore NMR signatures to slightly paramagnetic shifts compared to nonencapsulated molecules. The magnetic interactions reported here are robust phenomena independent of temperature and proportional to the applied magnetic field. The magnitude, tunability, and stability of the magnetic effects make the peapod nanomaterials potentially valuable for nanomagnetic shielding in nanoelectronics and nanobiomedical engineering.


CDK Substrate Phosphorylation and Ordering the Cell Cycle.
S phase and mitotic onset are brought about by the action of multiple different cyclin-CDK complexes. However, it has been suggested that changes in the total level of CDK kinase activity, rather than substrate specificity, drive the temporal ordering of S phase and mitosis. Here, we present a phosphoproteomics-based systems analysis of CDK substrates in fission yeast and demonstrate that the phosphorylation of different CDK substrates can be temporally ordered during the cell cycle by a single cyclin-CDK. This is achieved by rising CDK activity and the differential sensitivity of substrates to CDK activity over a wide dynamic range. This is combined with rapid phosphorylation turnover to generate clearly resolved substrate-specific activity thresholds, which in turn ensures the appropriate ordering of downstream cell-cycle events. Comparative analysis with wild-type cells expressing multiple cyclin-CDK complexes reveals how cyclin-substrate specificity works alongside activity thresholds to fine-tune the patterns of substrate phosphorylation.


BST2/Tetherin enhances entry of human cytomegalovirus.
Interferon-induced BST2/Tetherin prevents budding of vpu-deficient HIV-1 by tethering mature viral particles to the plasma membrane. BST2 also inhibits release of other enveloped viruses including Ebola virus and Kaposi's sarcoma associated herpesvirus (KSHV), indicating that BST2 is a broadly acting antiviral host protein. Unexpectedly however, recovery of human cytomegalovirus (HCMV) from supernatants of BST2-expressing human fibroblasts was increased rather than decreased. Furthermore, BST2 seemed to enhance viral entry into cells since more virion proteins were released into BST2-expressing cells and subsequent viral gene expression was elevated. A significant increase in viral entry was also observed upon induction of endogenous BST2 during differentiation of the pro-monocytic cell line THP-1. Moreover, treatment of primary human monocytes with siRNA to BST2 reduced HCMV infection, suggesting that BST2 facilitates entry of HCMV into cells expressing high levels of BST2 either constitutively or in response to exogenous stimuli. Since BST2 is present in HCMV particles we propose that HCMV entry is enhanced via a reverse-tethering mechanism with BST2 in the viral envelope interacting with BST2 in the target cell membrane. Our data suggest that HCMV not only counteracts the well-established function of BST2 as inhibitor of viral egress but also employs this anti-viral protein to gain entry into BST2-expressing hematopoietic cells, a process that might play a role in hematogenous dissemination of HCMV.


Regulation of 13(S)-hydroxyoctadecadienoic acid biosynthesis in Syrian hamster embryo fibroblasts by the epidermal growth factor receptor tyrosine kinase.
Metabolism of arachidonic and linoleic acid can be regulated by polypeptide growth factors in a variety of cell types. In Syrian hamster embryo (SHE) fibroblasts, epidermal growth factor (EGF) stimulates the conversion of exogenous linoleic acid to 13(S)-hydroxyoctadecadienoic acid (HODE). Inhibition of 13-HODE biosynthesis blocks the EGF-mitogenic response in SHE cells, and 13-HODE and its hydroperoxy precursor are potent and highly specific enhancers of EGF-dependent DNA synthesis. We demonstrated that EGF stimulates a biphasic production and release of endogenous 13-HODE. Through development of a stable isotope-dilution GC/MS assay for 13-HODE, we observed 13-HODE production as early as 5 min after EGF stimulation, and this initial phase peaked at 1 hr. A second rise in 13-HODE formation was seen at 2-4 hr, and this phase plateaued at 4-6 hr at a level of 30-40 ng/10(6) cells. EGF stimulation of 13-HODE biosynthesis is not mediated by transcriptional or translational regulation of the inducible form of prostaglandin H synthase. Based on enzyme inhibitor studies and structural characterization of products, the linoleate metabolite is apparently formed by an n-6 lipoxygenase that remains to be characterized. EGF stimulation of 13-HODE formation is linked with activation of the EGF receptor tyrosine kinase. Inhibition of EGF receptor tyrosine kinase activity with methyl-2,5-dihydroxycinnamate blocked EGF-dependent linoleic acid metabolism and EGF-regulated DNA synthesis. Potentiation of the EGF receptor tyrosine phosphorylation cascade through treatment of SHE cells with the tyrosine phosphatase inhibitor vanadate resulted in a 3-fold increase in EGF-stimulated 13-HODE production and a corresponding enhancement of the EGF mitogenic response. The coupling of EGF-regulated linoleic acid metabolism with the EGF receptor tyrosine kinase activity suggests the importance of specific linoleate compounds in mediating mitogenic signal transduction.


Correction to the plant canopy gap-size analysis theory used by the Tracing Radiation and Architecture of Canopies instrument.
A plant canopy gap-size analyzer, the Tracing Radiation and Architecture of Canopies (TRAC), developed by Chen and Cihlar [Appl Opt. 34, 6211(1995)] and commercialized and by 3rd Wave Engineering (Nepean, Canada), has been used around the world to quantify the fraction of photosynthetically activeradiation absorbed by plant canopies, the leaf area index (LAI), and canopy architectural parameters. The TRAC is walked under a canopy along transects to measure sunflecks that are converted into a gap-size distribution. A numerical gap-removal technique is performed to remove gaps that are not theoretically possible in a random canopy. The resulting reduced gap-size distribution is used to quantify the heterogeneity of the canopy and to improve LAI measurements. It is explicitly shown here that the original derivation of the clumping index was missing a normalization factor. For a very clumped canopy with a large gap faction, the resulting LAI can be more than 100% smaller than previously estimated. A test case is used to demonstrate that the new clumping index derivation allows a more accurate change of LAI to be measured.


Extraventricular neurocytomas: pathologic features and clinical outcome.
Neurocytic neoplasms usually arise within the lateral ventricles, generally as circumscribed, slowly growing masses curable by total resection. Both subtotal resection and histologic atypia are associated with an increased risk of recurrence. In contrast, neurocytic neoplasms situated within brain parenchyma, so-called "extraventricular neurocytomas" (EVNs), are not as well characterized. The relationships between histologic features and extent of resection versus clinical behavior have not been defined. We evaluated pathologic features, clinical data, and neuroimaging of 35 examples. The tumors occurred in 18 males and 17 females, age 5-76 years (median 34 years). All tumors involved the cerebrum. On imaging, EVNs were solitary, variably contrast-enhancing, and often (57%) cystic. Tumor cells were arranged in sheets, clusters, ribbons, or rosettes, in association with fine neuropil dispersed in broad zones that separated cell aggregates. Ganglion cell differentiation was seen in 66%. All tumors showed strong synaptophysin immunoreactivity. Despite the lack of apparent astrocytes in hematoxylin and eosin-stained sections, focal glial fibrillary acidic protein reactivity was seen in 46%. Eleven EVNs were designated "atypical" based on the presence of necrosis, vascular proliferation, or elevated mitotic activity (> or = 3 mitoses/10 high power fields). Nineteen tumors were subtotally resected or biopsied, whereas 14 were totally resected grossly. Seventeen patients underwent radiotherapy (mean 55 Gy). In 30 cases with follow-up, 10 tumors recurred, 3 causing death at 6, 14, and 43 months. All 10 recurrences followed subtotal resection. No totally resected tumors recurred. Thus, the majority of EVNs are well differentiated and appear unlikely to recur after gross total resection. Subtotal resection, atypical histologic features, and high cell proliferation rates correlate with recurrence.


Endogenous Pseudallescheria boydii endophthalmitis in a patient with ring-enhancing brain lesions.
A 46-year-old man, status post liver transplantation and taking immunosuppressive medications, was admitted after suffering a generalized seizure. Magnetic resonance imaging of the brain revealed two ring-enhancing lesions and treatment was begun for presumed toxoplasmic encephalitis. He was already receiving amphotericin B for a skin lesion suspected to be caused by candidiasis. One day after the seizure, he complained of photophobia in the left eye. Intraocular inflammation and a small infiltrate in the macula were seen. Vision deteriorated over the next three days. Vitreous tap and injection of amphotericin B and vancomycin were performed, but the intraocular inflammation continued to increase. On day 15, a vitrectomy was performed. The vitreous specimen ultimately grew Pseudallescheria boydii. The patient died on hospital day 30 from complications of the brain abscesses. Pseudallescheria boydii should be considered in the differential diagnosis of endophthalmitis, especially in patients with immunosuppression, serious medical disease, or ring-enhancing brain lesions.


Multi-tissue transcriptomes of caecilian amphibians highlight incomplete knowledge of vertebrate gene families.
RNA sequencing (RNA-seq) has become one of the most powerful tools to unravel the genomic basis of biological adaptation and diversity. Although challenging, RNA-seq is particularly promising for research on non-model, secretive species that cannot be observed in nature easily and therefore remain comparatively understudied. Among such animals, the caecilians (order Gymnophiona) likely constitute the least known group of vertebrates, despite being an old and remarkably distinct lineage of amphibians. Here, we characterize multi-tissue transcriptomes for five species of caecilians that represent a broad level of diversity across the order. We identified vertebrate homologous elements of caecilian functional genes of varying tissue specificity that reveal a great number of unclassified gene families, especially for the skin. We annotated several protein domains for those unknown candidate gene families to investigate their function. We also conducted supertree analyses of a phylogenomic dataset of 1,955 candidate orthologous genes among five caecilian species and other major lineages of vertebrates, with the inferred tree being in agreement with current views of vertebrate evolution and systematics. Our study provides insights into the evolution of vertebrate protein-coding genes, and a basis for future research on the molecular elements underlying the particular biology and adaptations of caecilian amphibians.


Kinetics study of the CN + CH4 hydrogen abstraction reaction based on a new ab initio analytical full-dimensional potential energy surface.
We have developed an analytical full-dimensional potential energy surface, named PES-2017, for the gas-phase hydrogen abstraction reaction between the cyano radical and methane. This surface is fitted using high-level ab initio information as input. Using the PES-2017 surface, a kinetics study was performed via two theoretical approaches: variational transition-state theory with multidimensional tunnelling (VTST-MT) and ring polymer molecular dynamics (RPMD). The results are compared with the experimental data. In the whole temperature range analysed, 300-1500 K, both theories agree within a factor of <2, reproducing the experimental behaviour taking into account the experimental uncertainties. At high temperatures, where the recrossing effects dominate and the RPMD theory is exact, both theories differ by a factor of about 20%; while at low temperatures this difference is larger, 45%. Note that in this temperature regime, the tunnelling effect is negligible. The CN + CH4/CD4 kinetic isotope effects are important, reproducing the scarce experimental evidence. The good agreement with the ab initio information used in the fitting process (self-consistency test) and with the kinetic behaviour in a wide temperature range gives confidence and strength to the new surface.


Synaptic plasticity in vitro: cell culture of identified Aplysia neurons mediating short-term habituation and sensitization.
The gill withdrawal reflex of the marine mollusk, Aplysia californica, shows habituation and sensitization, two simple forms of learning. In order to extend the cellular studies on synaptic plasticity underlying the changes in the reflex behavior, and to explore further the development of synaptic plasticity during synapse formation, we have sought to establish the neural circuit of the gill withdrawal reflex in vitro. We report here the reconstruction of the elementary gill withdrawal circuit in cell culture and find that the cells show short-term homosynaptic depression and heterosynaptic facilitation, the cellular mechanisms of habituation and sensitization, respectively.


How important are suppression of aldosterone and stimulation of atrial natriuretic peptide secretion in the natriuretic response to an acute sodium load in man?
1. Aldosterone is suppressed by sodium loading. We studied the contribution of this decrease in plasma aldosterone to the natriuresis after acute sodium loading in healthy volunteers. 2. Two litres of saline [0.9% (w/v) NaCl] were infused during the second hour of a 6 h infusion of aldosterone (3 pmol min-1 kg-1) or placebo in eight healthy young men. On the placebo day, plasma aldosterone decreased by 30 min after the start of saline infusion and remained suppressed. During aldosterone infusion, plasma aldosterone was maintained at around 400 pmol/l. 3. Urinary sodium excretion, lithium clearance and plasma atrial natriuretic peptide increased and plasma renin activity decreased after saline infusion, whether or not aldosterone was infused. However, from 60 to 240 min after saline infusion, natriuresis was significantly less during aldosterone infusion than on the placebo day. In addition, saline loading led to a progressive increase in the ratio of sodium clearance to lithium clearance, used as an index of the fractional distal tubular rejection of sodium, and in the ratio of urinary sodium to potassium. These increases were prevented by the infusion of aldosterone. 4. This study suggests that there are differences in the mechanisms determining the early and the later responses to an acute sodium load. Suppression of aldosterone may explain much of the later increase in natriuresis after saline infusion. In addition, the results are consistent with a role for atrial natriuretic peptide in the immediate increase in sodium excretion after saline loading.


Global and country underestimation of hepatocellular carcinoma (HCC) in 2012 and its implications.
The problems of screening costs, as well as poor data, potentially lead to the underestimation of the incidence of hepatocellular carcinoma (HCC). In particular, this is problematic in developing countries with limited resources and poor data. The study develops a model to inform policy makers of the true incidence and potential extra cost of HCC in a developing country context. Using Globocan 2012 data, we employed an ecological correlation design at country level to associate HCC incidence data with relevant determinant data like HBV-HCV and other exposure factors. A Poisson regression model was used to estimate potentially missed incident cases of HCC by country and region based on the country risk factor covariate values. The results indicated that HBV and HCV prevalence were significantly associated with HCC incidence (p<0.001) and potentially accounted for 94%% of incident HCC in 2012. We estimated a total of 120,772 potentially missed incident HCC cases in 2012. These cases are largely predicted for South Asia (>21,000), North Asia (>15,000), Western Africa (14,500) and Eastern Africa (12,500). Developing countries, with poorer quality data and a high historical burden of hepatitis, were predicted to have the majority of missed HCC cases in 2012 based on our model. These countries are, therefore, less able to detect, budget for or manage HCC. The high cost of HCC treatment, as well as its economic implications, poses a challenge in resource poor settings.


spn-F encodes a novel protein that affects oocyte patterning and bristle morphology in Drosophila.
The anteroposterior and dorsoventral axes of the Drosophila embryo are established during oogenesis through the activities of Gurken (Grk), a Tgfalpha-like protein, and the Epidermal growth factor receptor (Egfr). spn-F mutant females produce ventralized eggs similar to the phenotype produced by mutations in the grk-Egfr pathway. We found that the ventralization of the eggshell in spn-F mutants is due to defects in the localization and translation of grk mRNA during mid-oogenesis. Analysis of the microtubule network revealed defects in the organization of the microtubules around the oocyte nucleus. In addition, spn-F mutants have defective bristles. We cloned spn-F and found that it encodes a novel coiled-coil protein that localizes to the minus end of microtubules in the oocyte, and this localization requires the microtubule network and a Dynein heavy chain gene. We also show that Spn-F interacts directly with the Dynein light chain Ddlc-1. Our results show that we have identified a novel protein that affects oocyte axis determination and the organization of microtubules during Drosophila oogenesis.


General and erosive tooth wear of 16-year-old adolescents in Kuantan, Malaysia: prevalence and association with dental caries.
The objective of this study was to determine the prevalence and severity of general tooth wear (GTW), i.e. tooth wear irrespective of etiology and erosive tooth wear (ETW), i.e. tooth wear predominantly due to erosion; and also to investigate the relationship between ETW and dental caries experience in 16-year-old adolescents in Kuantan, Malaysia. A multi-staged cluster sampling method was employed. A total of 598 16-year-old adolescents participated in this study. Participants' demographic profile was assessed through a self-administered questionnaire. Clinical examinations were carried out under standardized conditions by a single examiner. The level of GTW was recorded using the modified Smith and Knight's Tooth Wear Index (TWI) whilst ETW were recorded using the Basic Erosive Wear Examination (BEWE) index. This index was developed to record clinical findings and assist in the decision-making process for the management of erosive tooth wear. Dental caries was recorded using the D3MFT index whereby D3 denotes obvious dental decay into dentine detected visually. The prevalence of GTW, ETW and dental caries, i.e. percentage of individuals found to have at least one lesion, was 99.8%, 45.0% and 27.8% respectively. Two thirds of affected teeth with GTW were observed to have a TWI score of 1 whereas almost all of the affected teeth with ETW had a BEWE score of 2. The mean D3MFT was 0.62 (95% CI 0.50, 0.73) with Decayed (D) teeth being the largest component, mean D3T was 0.36 (95% CI 0.30, 0.43). There was no significant association between socio-demographic factors and prevalence of ETW. Logistic regression analysis also showed no significant relationship between the prevalence of ETW and D3MFT (p > 0.05). Almost all adolescents examined had GTW but they were mainly early lesions. However, nearly half were found to have ETW of moderate severity (BEWE score 2). No significant relationship between the occurrence of erosive tooth wear and caries was observed in this population.


Pretreatment with coenzyme Q10 improves ovarian response and embryo quality in low-prognosis young women with decreased ovarian reserve: a randomized controlled trial.
Management of women with reduced ovarian reserve or poor ovarian response (POR) to stimulation is one of the major challenges in reproductive medicine. The primary causes of POR remain elusive and oxidative stress was proposed as one of the important contributors. It has been suggested that focus on the specific subpopulations within heterogeneous group of poor responders could assist in evaluating optimal management strategies for these patients. This study investigated the effect of anti-oxidant treatment with coenzyme Q10 (CoQ10) on ovarian response and embryo quality in young low-prognosis patients with POR. This prospective, randomized controlled study included 186 consecutive patients with POR stratified according to the POSEIDON classification group 3 (age < 35, poor ovarian reserve parameters). The participants were randomized to the CoQ10 pre-treatment for 60 days preceding IVF-ICSI cycle or no pre-treatment. The number of high quality embryos was a primary outcome measure. A total of 169 participants were evaluated (76 treated with CoQ10 and 93 controls); 17 women were excluded due to low compliance with CoQ10 administration. The baseline demographic and clinical characteristics were comparable between the groups. CoQ10 pretreatment resulted in significantly lower gonadotrophin requirements and higher peak E2 levels. Women in CoQ10 group had increased number of retrieved oocytes (4, IQR 2-5), higher fertilization rate (67.49%) and more high-quality embryos (1, IQR 0-2); p < 0.05. Significantly less women treated with CoQ10 had cancelled embryo transfer because of poor embryo development than controls (8.33% vs. 22.89%, p = 0.04) and more women from treatment group had available cryopreserved embryos (18.42% vs. 4.3%, p = 0.012). The clinical pregnancy and live birth rates per embryo transfer and per one complete stimulation cycle tended to be higher in CoQ10 group but did not achieve statistical significance. Pretreatment with CoQ10 improves ovarian response to stimulation and embryological parameters in young women with poor ovarian reserve in IVF-ICSI cycles. Further work is required to determine whether there is an effect on clinical treatment endpoints.


Farmers' Rights and Digital Sequence Information: Crisis or Opportunity to Reclaim Stewardship Over Agrobiodiversity?
Contestations about the way in which digital sequence information is used and regulated have created stumbling blocks across multiple international policy processes. Such schisms have profound implications for the way in which we manage and conceptualize agrobiodiversity and its benefits. This paper explores the relationship between farmers' rights, as recognized in the International Treaty on Plant Genetic Resources for Food and Agriculture, and the dematerialization of genetic resources. Using concepts of "stewardship" and "ownership" we emphasize the need to move away from viewing agrobiodiversity as a commodity that can be owned, toward a strengthened, proactive and expansive stewardship approach that recognizes plant genetic resources for food and agriculture as a public good which should be governed as such. Through this lens we analyze the relationship between digital sequence information and different elements of farmers' rights to compare and contrast implications for the governance of digital sequence information. Two possible parallel pathways are presented, the first envisaging an enhanced multilateral system that includes digital sequence information and which promotes and enhances the realization of farmers' rights; and the second a more radical approach that folds together concepts of stewardship, farmers' rights, and open source science. Farmers' rights, we suggest, may well be the linchpin for finding fair and equitable solutions for digital sequence information beyond the bilateral and transactional approach that has come to characterize access and benefit sharing under the Convention on Biological Diversity. Existing policy uncertainties could be seized as an unexpected but serendipitous opportunity to chart an alternative and visionary pathway for the rights of farmers and other custodians of plant genetic resources.


Influence of Ribavirin Serum Levels on Outcome of Antiviral Treatment and Anemia in Hepatitis C Virus Infection.
Ribavirin blood levels vary considerably between patients with standard weight-based dosing. Their impact on sustained virological response (SVR) with pegylated interferon and ribavirin is controversial, but has mostly been studied before the IL28b gene polymorphism as a possible confounder was discovered. The impact of serum ribavirin trough levels at week 4, at the end of treatment and of mean levels across the entire antiviral treatment with pegylated interferon and ribavirin on relapse, SVR rates and anemia was retrospectively studied by univariate and multivariable logistic regression analyses in 214 patients with HCV genotype 1-4 infection, including 88 patients with available IL28b genotyping. Mean ribavirin levels varied between 0.68-5.65 mg/l and significantly differed between patients with or without SVR. By multivariable regression including age, sex, HCV viral load, HCV genotype, liver fibrosis stage, prior treatments, immunosuppression and IL28b genotype, ribavirin levels consistently displayed significant influence on SVR and relapse without indication for a specific importance of higher concentrations early or late in the treatment course. Although hemoglobin decline was on average more pronounced in patients with higher ribavirin levels, hemoglobin remained relatively stable in a significant proportion of these, indicating that ribavirin levels alone are insufficient to predict anemia. While data are scarce to draw conclusions applicable for modern DAA therapies, these results support ribavirin treatment based on serum levels instead of purely weight-based dosing in combination with pegylated interferon.


Dental development in living and fossil orangutans.
Numerous studies have investigated molar development in extant and fossil hominoids, yet relatively little is known about orangutans, the only great ape with an extensive fossil record. This study characterizes aspects of dental development, including cuspal enamel daily secretion rate, long-period line periodicities, cusp-specific molar crown formation times and extension rates, and initiation and completion ages in living and fossil orangutan postcanine teeth. Daily secretion rate and periodicities in living orangutans are similar to previous reports, while crown formation times often exceed published values, although direct comparisons are limited. One wild Bornean individual died at 4.5 years of age with fully erupted first molars (M1s), while a captive individual and a wild Sumatran individual likely erupted their M1s around five or six years of age. These data underscore the need for additional samples of orangutans of known sex, species, and developmental environment to explore potential sources of variation in molar emergence and their relationship to life history variables. Fossil orangutans possess larger crowns than living orangutans, show similarities in periodicities, and have faster daily secretion rate, longer crown formation times, and slower extension rates. Molar crown formation times exceed reported values for other fossil apes, including Gigantopithecus blacki. When compared to African apes, both living and fossil orangutans show greater cuspal enamel thickness values and periodicities, resulting in longer crown formation times and slower extension rates. Several of these variables are similar to modern humans, representing examples of convergent evolution. Molar crown formation does not appear to be equivalent among extant great apes or consistent within living and fossil members of Pongo or Homo.