Does ethnicity influence dementia, stroke and mortality risk? Evidence from the UK Biobank.
The number of people with dementia and stroke is increasing worldwide. There is increasing evidence that there are clinically relevant genetic differences across ethnicities. This study aims to quantify risk factors of dementia, stroke, and mortality in Asian and black participants compared to whites. 272,660 participants from the UK Biobank were included in the final analysis, among whom the vast majority are white (n = 266,671, 97.80%), followed by Asian (n = 3,790, 1.35%), and black (n = 2,358, 0.84%) participants. Cumulative incidence risk was calculated based on all incident cases occurring during the follow-up of the individuals without dementia and stroke at baseline. We compared the allele frequency of variants in Asian and black participants with the referent ethnicity, whites, by chi-square test. Hierarchical cluster analysis was used in the clustering analysis. Significance level corrected for the false discovery rate was considered. After adjusting for risk factors, black participants have an increased risk of dementia and stroke compared to white participants, while Asians has similar odds to the white. The risk of mortality is not different in blacks and white participants but Asians have a decreased risk. The study provides important insights into the potential differences in the risk of dementia and stroke among different ethnic groups. Specifically, the study found that black individuals had a higher incidence of dementia and stroke compared to white individuals living in the UK. These findings are particularly significant as they suggest that there may be underlying factors that contribute to these differences, including genetic, environmental, and social factors. By identifying these differences, the study helps to inform interventions and policies aimed at reducing the risk of dementia and stroke, particularly among high-risk populations.


Evaluation of Vertebrate-Specific Replication-Defective Zika Virus, a Novel Single-Cycle Arbovirus Vaccine, in a Mouse Model.
The flavivirus Zika (ZIKV) has emerged as a global threat, making the development of a ZIKV vaccine a priority. While live-attenuated vaccines are known to induce long-term immunity but reduced safety, inactivated vaccines exhibit a weaker immune response as a trade-off for increased safety margins. To overcome the trade-off between immunogenicity and safety, the concept of a third-generation flavivirus vaccine based on single-cycle flaviviruses has been developed. These third-generation flavivirus vaccines have demonstrated extreme potency with a high level of safety in animal models. However, the production of these single-cycle, encapsidation-defective flaviviruses requires a complicated virion packaging system. Here, we investigated a new single-cycle flavivirus vaccine, a vertebrate-specific replication-defective ZIKV (VSRD-ZIKV), in a mouse model. VSRD-ZIKV replicates to high titers in insect cells but can only initiate a single-round infection in vertebrate cells. During a single round of infection, VSRD-ZIKV can express all the authentic viral antigens in vertebrate hosts. VSRD-ZIKV immunization elicited a robust cellular and humoral immune response that protected against a lethal ZIKV challenge in AG129 mice. Additionally, VSRD-ZIKV-immunized pregnant mice were protected against vertically transferring a lethal ZIKV infection to their offspring. Immunized male mice were protected and prevented viral accumulation in the testes after being challenged with lethal ZIKV. Overall, our results indicate that VSRD-ZIKV induces a potent protective immunity against ZIKV in a mouse model and represents a promising approach to develop novel single-cycle arbovirus vaccines.


Dark oxidation of dissolved and liquid elemental mercury in aquatic environments.
Elemental mercury (Hg0) can be found in liquid or dissolved forms in aquatic systems. Whereas dissolved Hg0 is measured in virtually all aquatic systems, liquid Hg0 droplets are mainly observed at poorly lit sediment/water interfaces of ecosystems with local point sources such as hydro-thermal vents, gold extraction sites, and near industrial facilities. Here, we report that, in the dark, liquid and dissolved forms of Hg behave differently with respect to their oxidation. Liquid Hg0 is rapidly oxidized in oxygenated solution in the presence of chloride. Liquid Hg0 oxidation rates are positively correlated with chloride concentrations and droplet surface area. When liquid Hg is removed from solution, the oxidation stops even though the solution is still saturated with dissolved Hg0. Liquid Hg0 droplets in oxygenated marine or brackish environments should be oxidized and release Hg2+ to solution. In freshwaters or anoxic seawater, liquid Hg will dissolve releasing Hg(aq)0 which, itself, will slowly oxidize.


Comprehensive Omics Analysis of a Novel Small-Molecule Inhibitor of Chemoresistant Oncogenic Signatures in Colorectal Cancer Cell with Antitumor Effects.
Tumor recurrence from cancer stem cells (CSCs) and metastasis often occur post-treatment in colorectal cancer (CRC), leading to chemoresistance and resistance to targeted therapy. MYC is a transcription factor in the nuclei that modulates cell growth and development, and regulates immune response in an antitumor direction by mediating programmed death ligand 1 (PD-L1) and promoting CRC tumor recurrence after adjuvant chemotherapy. However, the molecular mechanism through which c-MYC maintains stemness and confers treatment resistance still remains elusive in CRC. In addition, recent reports demonstrated that CRC solid colon tumors expresses C-X-C motif chemokine ligand 8 (CXCL8). Expression of CXCL8 in CRC was reported to activate the expression of PD-L1 immune checkpoint through c-MYC, this ultimately induces chemoresistance in CRC. Accumulating studies have also demonstrated increased expression of CXCL8, matrix metalloproteinase 7 (MMP7), tissue inhibitor of metalloproteinase 1 (TIMP1), and epithelial-to-mesenchymal transition (EMT) components, in CRC tumors suggesting their potential collaboration to promote EMT and CSCs. TIMP1 is MMP-independent and regulates cell development and apoptosis in various cancer cell types, including CRC. Recent studies showed that TIMP1 cleaves CXCL8 on its chemoattractant, thereby influencing its mechanistic response to therapy. This therefore suggests crosstalk among the c-MYC/CXCL8/TIMP1 oncogenic signatures. In this study, we explored computer simulations through bioinformatics to identify and validate that the MYC/CXCL8/TIMP1 oncogenic signatures are overexpressed in CRC, Moreover, our docking results exhibited putative binding affinities of the above-mentioned oncogenes, with our novel small molecule, RV59, Finally, we demonstrated the anticancer activities of RV59 against NCI human CRC cancer cell lines both as single-dose and dose-dependent treatments, and also demonstrated the MYC/CXCL8/TIMP1 signaling pathway as a potential RV59 drug target.


Cost-Effectiveness Analysis of The Real Cost Campaign's Effect on Smoking Prevention.
A previous study found that the Food and Drug Administration's The Real Cost national tobacco education campaign was associated with preventing approximately 350,000 U.S. youth from initiating smoking between 2014 and 2016. This study translates the reduction in smoking initiation into monetary terms by examining the cost effectiveness of the campaign. The cost effectiveness of The Real Cost was assessed by measuring efficiency in two ways: (1) estimating the cost per quality-adjusted life year saved and (2) estimating the total monetary return on investment by comparing the cost savings associated with the campaign to campaign expenditures. Analyses were conducted in 2017. The Real Cost averted an estimated 175,941 youth from becoming established smokers between 2014 and 2016. Campaign expenditures totaled $246,915,233. The cost per quality-adjusted life year saved of the campaign was $1,337. When considering the costs of smoking, the averted established smokers represent >$31 billion in cost savings ($1.3 billion when only external costs considered). The overall return on investment of the campaign was $128 in cost savings for every $1 spent ($4 for every $1 spent when only external costs considered). These conclusions were robust to sensitivity analyses surrounding the parameters. Campaign expenditures were cost efficient. The cost savings resulting from The Real Cost represent a large reduction in the financial burden to individuals, their families, and society as a result of tobacco. Public health campaigns, like The Real Cost, that reduce tobacco-related morbidity and mortality for a generation of U.S. youth also provide substantial cost savings.


Is there more dementia, depression and neurosis in New York? A comparative study of the elderly in New York and London using the computer diagnosis AGECAT.
A computerised diagnosis, AGECAT, is applied to data from random community samples of the elderly in New York and London in order to examine the prevalence of mental illness in the two cities, especially the result reported from the same study, using a different diagnostic procedure, that the proportion of dementia was higher in New York. The greater proportion of dementia in New York is confirmed for all AGECAT's levels of diagnostic confidence, for both sexes and at all half-decades over 65 years. Provided that this difference is upheld and genetic differences can be discounted, these findings would seem to support a contributory environmental cause for dementia. Proportions of depression as a whole are similar in both cities. Syndrome case and sub-case levels of neurotic disorder are generally higher in London.


Instrumentation and design of a frequency-domain diffuse optical tomography imager for breast cancer detection.
The instrument development and design of a prototype frequency-domain optical imaging device for breast cancer detection is described in detail. This device employs radio-frequency intensity modulated near-infrared light to image quantitatively both the scattering and absorption coefficients of tissue. The functioning components of the system include a laser diode and a photomultiplier tube, which are multiplexed automatically through 32 large core fiber optic bundles using high precision linear translation stages. Image reconstruction is based on a finite element solution of the diffusion equation. This tool for solving the forward problem of photon migration is coupled to an iterative optical property estimation algorithm, which uses a Levenberg-Marquardt routine with total variation minimization. The result of this development is an automated frequency-domain optical imager for computed tomography which produces quantitatively accurate images of the test phantoms used to date. This paper is a description and characterization of an automated frequency-domain computed tomography scanner, which is more quantitative than earlier systems used in diaphanography because of the combination of intensity modulated signal detection and iterative image reconstruction.


Asthma inhaler adherence determinants in adults: systematic review of observational data.
Nonadherence to inhaled medication leads to poor asthma control and increased healthcare utilisation. Many studies exploring adherence determinants have been conducted, but summaries of the evidence are scarce. We performed a systematic review of observational research on determinants of asthma inhaler adherence among adults. We searched for articles in English reporting quantitative observational studies on inhaler adherence correlates among adults in developed countries, published in EMBASE, Medline, PsychInfo and PsychArticles in 1990-2014. Two coders independently assessed eligibility and extracted data, and assessed study quality. Results were summarised qualitatively into social and economic, and healthcare-, therapy-, condition- and patient-related factors. The 51 studies included mainly examined patient-related factors and found consistent links between adherence and stronger inhaler-necessity beliefs, and possibly older age. There was limited evidence on the relevance of other determinants, partly due to study heterogeneity regarding the types of determinants examined. Methodological quality varied considerably and studies performed generally poorly on their definitions of variables and measures, risk of bias, sample size and data analysis. A broader adoption of common methodological standards and health behaviour theories is needed before cumulative science on the determinants of adherence to asthma inhalers among adults can develop further.


A Bayesian approach to the stereo correspondence problem.
I present a probabilistic approach to the stereo correspondence problem. Rather than trying to find a single solution in which each point in the left retina is assigned a partner in the right retina, all possible matches are considered simultaneously and assigned a probability of being correct. This approach is particularly suitable for stimuli where it is inappropriate to seek a unique partner for each retinal position--for instance, where objects occlude each other, as in Panum's limiting case. The probability assigned to each match is based on a Bayesian analysis previously developed to explain psychophysical data (Read, 2002). This provides a convenient way to incorporate constraints that enable the ill-posed correspondence problem to be solved. The resulting model behaves plausibly for a variety of different stimuli.


Protection of mouse retinal ganglion cell axons and soma from glaucomatous and ischemic injury by cytoplasmic overexpression of Nmnat1.
The Wlds mutation affords protection of retinal ganglion cell (RGC) axons in retinal ischemia and in inducible and hereditary preclinical models of glaucoma. We undertook the present study to determine whether the Nmnat1 portion of the chimeric protein provides axonal and somatic protection of RGCs in models of ischemia and glaucoma, particularly when localized to nonnuclear regions of the cell. The survival and integrity of RGC axons and soma from transgenic mice with confirmed cytoplasmic overexpression of Nmnat1 in retina and optic nerve (cytNmnat1-Tg mice) were examined in the retina and postlaminar optic nerve 4 days following acute retinal ischemia, and 3 weeks following the chronic elevation of intraocular pressure. Ischemia- and glaucoma-induced disruptions of proximal segments of RGC axons that comprise the nerve fiber layer in wild-type mice were both robustly abrogated in cytNmnat1-Tg mice. More distal portions of RGC axons within the optic nerve were also protected from glaucomatous disruption in the transgenic mice. In both disease models, Nmnat1 overexpression in extranuclear locations significantly enhanced the survival of RGC soma. Overexpression of Nmnat1 in the cytoplasm and axons of RGCs robustly protected against both ischemic and glaucomatous loss of RGC axonal integrity, as well as loss of RGC soma. These findings reflect the more pan-cellular protection of CNS neurons that is realized by cytoplasmic Nmnat1 expression, and thus provide a therapeutic strategy for protecting against retinal neurodegeneration, and perhaps other CNS neurodegenerative diseases as well.


Group versus individual cognitive treatment for obsessive-compulsive disorder: Changes in severity at post-treatment and one-year follow-up.
Very few studies have compared the efficacy of individual and group cognitive behaviour therapy (CBT) for the treatment of Obsessive-Compulsive Disorder (OCD) by taking into consideration the change in OCD severity in both the short and long term. To conduct an open trial of individual versus group CBT for OCD, comparing the clinical and statistically significant changes in severity both at post-treatment and one year later. Forty-two OCD subjects were assigned to individual (n = 18) or group CBT (n = 24, in four groups). Sixteen and 22 subjects completed the treatment in the individual and group conditions, respectively. The Yale-Brown Obsessive Compulsive Scale was recorded at pre-treatment, post-treatment and at the one-year follow-up. At the end of treatment, the clinically significant change was comparable for the two treatment conditions and remained stable at the one-year follow-up. Of the 16 participants who completed the individual CBT treatment, 68.75% were classified as recovered at post-treatment, compared to 40.9% of those receiving group CBT. At follow-up the rate of recovery decreased to 62.5% in individual CBT and to 31.8% in group CBT. Group CBT is effective in decreasing OCD severity. The post-treatment changes were maintained one year later. Nevertheless, these changes were higher in the individual delivery of CBT.


Delay-related cerebral activity and motor preparation.
Flexible goal-oriented behavior requires the ability to carry information across temporal delays. This ability is associated with sustained neural firing. In cognitive terms, this ability has often been associated with the maintenance of sensory material online, as during short-term memory tasks, or with the retention of a motor code, as during movement preparation tasks. The general issue addressed in this paper is whether short-term storage of sensory information and preparation of motor responses rely on different anatomical substrates. We used functional magnetic resonance imaging (fMRI) to measure sustained and time-varying delay-related cerebral activity evoked during performance of a delay non-match to sample (DNMS) task, where task contingencies rather than explicit instructions ensured that either sensory or motor representations were used to cross the delay period on each trial. This approach allowed us to distinguish sensory from motor characteristics of delay-related activity evoked by task contingencies, rather than differences in the control of short-term storage driven by verbal instructions. Holding sensory material online evoked both sustained and time-varying delay-related activity in prefrontal regions, whereas movement preparation evoked delay-related responses in precentral areas. Intraparietal cortex was sensitive to the presence of memoranda, but indifferent to the type of information that was retained in memory. Our findings indicate that short-term storage of sensory information and preparation of motor responses rely on partially segregated cerebral circuits. In the frontal lobe, these circuits are organized along a rostro-caudal dimension, corresponding to the sensory or motor nature of the stored material.


Undersea compound radiometer.
We have built a simple, undersea radiometer that measures ten integral moments of the radiance as functions of depth in natural waters. From these data it is possible to calculate nine spherical moments of the scattering function, provided that this function varies slowly in the horizontal planes (i.e., the water is fairly stratified). This technique inverts the equations of radiative transfer, which avoids some of the limitations of conventional instruments. We took the instrument on a voyage in the coastal waters of San Diego and were able to measure the absorption coefficient in real time and in situ; we have been able to recover scattering functions with the help of nearly concurrent attenuation measurements.


Videolaryngoscopy vs. Direct Laryngoscopy for Elective Airway Management in Paediatric Anaesthesia: A prospective randomised controlled trial.
The indirect visualisation of the glottic area with a videolaryngoscope could improve intubation conditions which may possibly lead to a higher success rate of the first intubation attempt. Comparison of videolaryngoscopy and direct laryngoscopy for elective airway management in paediatric patients. Prospective randomised controlled trial. Operating room. 535 paediatric patients undergoing elective anaesthesia with tracheal intubation. 501 patients were included in the final analysis. Patients were randomly allocated to the videolaryngoscopy group (n = 265) and to the direct laryngoscopy group (n = 269) for the primary airway management. The first attempt intubation success rate was assessed as the primary outcome. The secondary outcomes were defined as: the time to successful intubation (time to the first EtCO2 wave), the overall intubation success rate, the number of intubation attempts, the incidence of complications, and the impact of the length of the operator's clinical practice. The study was terminated after the planned interim analysis for futility. There were no significant demographic differences between the two groups. The first attempt intubation success rate was lower in the videolaryngoscopy group; 86.8% (n = 211) vs. 92.6% (n = 239), P = 0.046. The mean time to the first EtCO2 wave was longer in the videolaryngoscopy group at 39.0 s ± 36.7 compared to the direct laryngoscopy group, 23.6 s ± 24.7 (P < 0.001). There was no difference in the overall intubation success rate, in the incidence of complications nor significant difference based on the length of the clinical practice of the operator. The first attempt intubation success rate was lower in the videolaryngoscopy group in comparison to the direct laryngoscopy group. The time needed for successful intubation with videolaryngoscopy was longer compared with direct laryngoscopy. Clinicaltrials.gov identifier: NCT03747250.


Mental health outcomes associated with profiles of risk and resilience among U.S. Army spouses.
The current study examined patterns of risk and protective factors among military families and associations with mental health diagnoses among U.S. Army spouses. Spouses (N = 3,036) completed a survey of family psychosocial fitness, which informed protective factors including coping, family cohesion, and social support. Survey results were linked with Department of Defense archival data, which provided information on military-specific risks, including relocation, deployments, and reunification, as well as mental health care diagnoses. The three-step method of latent profile analysis identified six profiles, suggesting significant heterogeneity in military families with respect to their access to resources and exposure to risk. The largest profile of families (40.48% of the sample) had limited risk exposure and considerable strengths. Variability in risk and protection across profiles was associated with statistically significant differences in the prevalence of mental health diagnoses among spouses (χ² = 108.968, df = 5, p < .001). The highest prevalence of mental health diagnoses among Army spouses (41.2%) was observed in the profile with the lowest levels of protective factors. Findings point to the importance of evaluating both concurrent risk and protective factors. Increasing access to resources may be a fruitful avenue for prevention among military families that are struggling. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Dietary fish oil modulates macrophage fatty acids and decreases arthritis susceptibility in mice.
B10.RIII and B10.G mice were transferred from a diet of laboratory rodent chow to a standard diet in which all the fat (5% by weight) was supplied as either fish oil (17% eicosapentaenoic acid [EPA], 12% docosahexaenoic acid [DHA], 0% arachidonic acid [AA], and 2% linoleic acid) or corn oil (0% EPA, 0% DHA, 0% AA, and 65% linoleic acid). The fatty acid composition of the macrophage phospholipids from mice on the chow diet was similar to that of mice on a corn oil diet. Mice fed the fish oil diet for only 1 wk showed substantial increases in macrophage phospholipid levels of the omega-3 fatty acids (of total fatty acid 4% was EPA, 10% docosapentaenoic acid [DPA], and 10% DHA), and decreases in omega-6 fatty acids (12% was AA, 2% docosatetraenoic acid [DTA], and 4% linoleic acid) compared to corn oil-fed mice (0% EPA, 0% DPA, 6% DHA, 20% AA, 9% DTA, and 8% linoleic acid). After 5 wk this difference between the fish oil-fed and corn oil-fed mice was even more pronounced. Further small changes occurred at 5-9 wk. We studied the prostaglandin (PG) and thromboxane (TX) profile of macrophages prepared from mice fed the two diets just before being immunized with collagen. Irrespective of diet, macrophages prepared from female mice and incubated for 24 h had significantly more PG and TX in the medium than similarly prepared macrophages from male mice. The increased percentage of EPA and decreased percentage of AA in the phospholipids of the macrophages prepared from the fish oil-fed mice was reflected in a reduction in the amount of PGE2 and PGI2 in the medium relative to identically incubated macrophages prepared from corn oil-fed mice. When this same fish oil diet was fed to B10.RIII mice for 26 d before immunization with type II collagen, the time of onset of arthritis was increased, and the incidence and severity of arthritis was reduced compared to arthritis induced in corn oil-fed mice. The females, especially those on the fish oil diet, tended to have less arthritis than the males. These alterations in the fatty acid pool available for PG and leukotriene synthesis suggest a pivotal role for the macrophage and PG in the immune and/or inflammatory response to type II collagen.


The rhomboid-like superfamily: molecular mechanisms and biological roles.
The rhomboid proteases were first discovered as regulators of Drosophila EGF receptor signaling; soon after, it was recognized that they represented the founder members of a widespread family of intramembrane serine proteases conserved in all kingdoms. More recently still, the family was promoted to a superfamily, encompassing a wide variety of distantly related proteins. One of the surprises has been that many members of the rhomboid-like superfamily are not active proteases. Given the size of this clan, and its relatively recent discovery, there is still much to learn. Nevertheless, we already understand much about how rhomboid proteases perform their surprising function of cleaving transmembrane domains. We also already know that members of the rhomboid-like superfamily participate in biological functions as diverse as growth factor signaling, mitochondrial dynamics, inflammation, parasite invasion, and the machinery of protein quality control. Their potential medical significance is now becoming apparent in several areas.


Whole-genome-based phylogenomic analysis of the Belgian 2016-2017 influenza A(H3N2) outbreak season allows improved surveillance.
Seasonal influenza epidemics are associated with high mortality and morbidity in the human population. Influenza surveillance is critical for providing information to national influenza programmes and for making vaccine composition predictions. Vaccination prevents viral infections, but rapid influenza evolution results in emerging mutants that differ antigenically from vaccine strains. Current influenza surveillance relies on Sanger sequencing of the haemagglutinin (HA) gene. Its classification according to World Health Organization (WHO) and European Centre for Disease Prevention and Control (ECDC) guidelines is based on combining certain genotypic amino acid mutations and phylogenetic analysis. Next-generation sequencing technologies enable a shift to whole-genome sequencing (WGS) for influenza surveillance, but this requires laboratory workflow adaptations and advanced bioinformatics workflows. In this study, 253 influenza A(H3N2) positive clinical specimens from the 2016-2017 Belgian season underwent WGS using the Illumina MiSeq system. HA-based classification according to WHO/ECDC guidelines did not allow classification of all samples. A new approach, considering the whole genome, was investigated based on using powerful phylogenomic tools including beast and Nextstrain, which substantially improved phylogenetic classification. Moreover, Bayesian inference via beast facilitated reassortment detection by both manual inspection and computational methods, detecting intra-subtype reassortants at an estimated rate of 15 %. Real-time analysis (i.e. as an outbreak is ongoing) via Nextstrain allowed positioning of the Belgian isolates into the globally circulating context. Finally, integration of patient data with phylogenetic groups and reassortment status allowed detection of several associations that would have been missed when solely considering HA, such as hospitalized patients being more likely to be infected with A(H3N2) reassortants, and the possibility to link several phylogenetic groups to disease severity indicators could be relevant for epidemiological monitoring. Our study demonstrates that WGS offers multiple advantages for influenza monitoring in (inter)national influenza surveillance, and proposes an improved methodology. This allows leveraging all information contained in influenza genomes, and allows for more accurate genetic characterization and reassortment detection.


Hedgehog Pathway Inhibition Hampers Sphere and Holoclone Formation in Rhabdomyosarcoma.
Rhabdomyosarcoma (RMS) is the most common type of soft tissue sarcoma in children and can be divided into two main subtypes: embryonal (eRMS) and alveolar (aRMS). Among the cellular heterogeneity of tumors, the existence of a small fraction of cells called cancer stem cells (CSC), thought to be responsible for the onset and propagation of cancer, has been demonstrated in some neoplasia. Although the existence of CSC has been reported for eRMS, their existence in aRMS, the most malignant subtype, has not been demonstrated to date. Given the lack of suitable markers to identify this subpopulation in aRMS, we used cancer stem cell-enriched supracellular structures (spheres and holoclones) to study this subpopulation. This strategy allowed us to demonstrate the capacity of both aRMS and eRMS cells to form these structures and retain self-renewal capacity. Furthermore, cells contained in spheres and holoclones showed significant Hedgehog pathway induction, the inhibition of which (pharmacologic or genetic) impairs the formation of both holoclones and spheres. Our findings point to a crucial role of this pathway in the maintenance of these structures and suggest that Hedgehog pathway targeting in CSC may have great potential in preventing local relapses and metastases.


Molecular biomarkers in multiple sclerosis.
Multiple sclerosis (MS) is an inflammatory-neurodegenerative disease of the central nervous system presenting with significant inter- and intraindividual heterogeneity. However, the application of clinical and imaging biomarkers is currently not able to allow individual characterization and prediction. Complementary, molecular biomarkers which are easily quantifiable come from the areas of immunology and neurobiology due to the causal pathomechanisms and can excellently complement other disease characteristics. Only a few molecular biomarkers have so far been routinely used in clinical practice as their validation and transfer take a long time. This review describes the characteristics that an ideal MS biomarker should have and the challenges of establishing new biomarkers. In addition, clinically relevant and promising biomarkers from the blood and cerebrospinal fluid are presented which are useful for MS diagnosis and prognosis as well as for the assessment of therapy response and side effects.


Combined use of Bispectral Index and A-Line Autoregressive Index to assess anti-nociceptive component of balanced anaesthesia during lumbar arthrodesis.
This study evaluated the A-Line Autoregressive Index (AAI) response to surgical stimulation during lumbar arthrodesis, as an estimate of the anti-nociceptive component of a Bispectral Index (BIS) guided anaesthesia combined with epidural analgesia. An epidural catheter was inserted in 23 patients allocated randomly to receive ropivacaine plus clonidine (Group R) or normal saline (Group S) epidurally. General anaesthesia was induced with propofol, cis-atracurium and a remifentanil infusion that was stopped 3 min after tracheal intubation, and maintained using sevoflurane to keep BIS at 50 (range 40-60). Mean arterial pressure, heart rate, end-tidal sevoflurane, BIS and AAI were analysed from 2 min before to 17 min after surgical incision. While BIS was maintained at 50, AAI significantly increased from a 2 min averaged value of 12 (4) to 21 (7) in Group S within the first 5 min after surgical incision, but did not change in Group R. Maximum AAI values reached during the study period were significantly higher in Group S than in Group R [38 (12) and 27 (10), respectively]. Binary logistic regression analysis allowed the calculation of AAI threshold values above which the probability of predominant nociception over anti-nociception was higher than 95%. At 1 MAC sevoflurane concentration, a 2 min averaged AAI of 35 or an AAI peak value of 62 were associated with such a probability. During a BIS-guided constant level of hypnosis, AAI response to the onset of surgical stimulation significantly differs according to the analgesic regimen. Further studies are needed to refine the estimation of sensitivity and specificity of this variable in assessing the balance between nociception and anti-nociception during general anaesthesia.


Proton pump inhibitors: impact on glucose metabolism.
Diabetes mellitus is a complex chronic disease associated with an absolute insulin deficiency in type 1 diabetes (T1D) and a progressive deterioration of β-cell function in type 2 diabetes (T2D). T2D pathophysiology has numerous defects including incretin deficiency/resistance. Gastrin has demonstrated to be an islet growth factor (like glucagon-like peptide-1, epidermal growth factor, transforming growth factor-α,…) and be able to restore a functional β-cell mass in diabetic animals. This hormone is likely to stimulate insulin secretion during an ordinary protein-rich meal, this is, to have an incretin-like effect. Proton pump inhibitors (PPIs) can raise serum gastrin concentration significantly and therefore, affect to glucose metabolism through promoting β-cell regeneration/expansion and also enhancing insulin secretion. The present paper aims to review studies concerning the effect of PPIs on glucose metabolism. Several research groups have recently explored the potential role of this class of drugs on glycemic control, mainly in T2D. The results show antidiabetic properties for the PPIs with a global glucose-lowering power around 0.6-0.7 % points of HbA1c, but the level of evidence for the available literature is still not high. If these data start to become demonstrated in the ongoing clinical trials, PPIs could become a new antidiabetic agent with a good and safe profile for T2D and even useful for T1D, particularly in the area of islet transplantation to preserve β-cell mass.


Pyricularia graminis-tritici, a new Pyricularia species causing wheat blast.
Pyricularia oryzae is a species complex that causes blast disease on more than 50 species of poaceous plants. Pyricularia oryzae has a worldwide distribution as a rice pathogen and in the last 30 years emerged as an important wheat pathogen in southern Brazil. We conducted phylogenetic analyses using 10 housekeeping loci for 128 isolates of P. oryzae sampled from sympatric populations of wheat, rice, and grasses growing in or near wheat fields. Phylogenetic analyses grouped the isolates into three major clades. Clade 1 comprised isolates associated only with rice and corresponds to the previously described rice blast pathogen P. oryzae pathotype Oryza (PoO). Clade 2 comprised isolates associated almost exclusively with wheat and corresponds to the previously described wheat blast pathogen P. oryzae pathotype Triticum (PoT). Clade 3 contained isolates obtained from wheat as well as other Poaceae hosts. We found that Clade 3 is distinct from P. oryzae and represents a new species, Pyricularia graminis-tritici (Pgt). No morphological differences were observed among these species, but a distinctive pathogenicity spectrum was observed. Pgt and PoT were pathogenic and highly aggressive on Triticum aestivum (wheat), Hordeum vulgare (barley), Urochloa brizantha (signal grass), and Avena sativa (oats). PoO was highly virulent on the original rice host (Oryza sativa), and also on wheat, barley, and oats, but not on signal grass. We conclude that blast disease on wheat and its associated Poaceae hosts in Brazil is caused by multiple Pyricularia species. Pyricularia graminis-tritici was recently found causing wheat blast in Bangladesh. This indicates that P. graminis-tritici represents a serious threat to wheat cultivation globally.


Advances in immunotherapy for hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is a prevalent disease with a progression that is modulated by the immune system. Systemic therapy is used in the advanced stage and until 2017 consisted only of antiangiogenic tyrosine kinase inhibitors (TKIs). Immunotherapy with checkpoint inhibitors has shown strong anti-tumour activity in a subset of patients and the combination of the anti-PDL1 antibody atezolizumab and the VEGF-neutralizing antibody bevacizumab has or will soon become the standard of care as a first-line therapy for HCC, whereas the anti-PD1 agents nivolumab and pembrolizumab are used after TKIs in several regions. Other immune strategies such as adoptive T-cell transfer, vaccination or virotherapy have not yet demonstrated consistent clinical activity. Major unmet challenges in HCC checkpoint immunotherapy are the discovery and validation of predictive biomarkers, advancing treatment to earlier stages of the disease, applying the treatment to patients with liver dysfunction and the discovery of more effective combinatorial or sequential approaches. Combinations with other systemic or local treatments are perceived as the most promising opportunities in HCC and some are already under evaluation in large-scale clinical trials. This Review provides up-to-date information on the best use of currently available immunotherapies in HCC and the therapeutic strategies under development.


Identification of prognostic biomarker in predicting hepatocarcinogenesis from cirrhotic liver using protein and gene signatures.
Cirrhosis primes the liver for hepatocellular carcinoma (HCC) development. However, biomarkers that predict HCC in cirrhosis patients are lacking. Thus, we aimed to identify a biomarker directly from protein analysis and relate it with transcriptomic data to validate in larger cohorts. Forty-six patients who underwent hepatectomy for HCC that arose from cirrhotic liver were enrolled. Reverse-phase protein array and microarray data of these patients were analyzed. Clinical validation was performed in two independent cohorts and functional validation using cell and tissue microarray (TMA). Systematic analysis performed after selecting 20 proteins from 201 proteins with AUROC >70 effectively categorized patients into high (n = 20) or low (n = 26) risk HCC groups. Proteome-derived late recurrence (PDLR)-gene signature comprising 298 genes that significantly differed between high and low risk groups predicted HCC well in a cohort of 216 cirrhosis patients and also de novo HCC recurrence in a cohort of 259 patients who underwent hepatectomy. Among 20 proteins that were selected for analysis, caveolin-1 (CAV1) was the most dominant protein that categorized the patients into high and low risk groups (P < .001). In a multivariate analysis, compared with other clinical variables, the PDLR-gene signature remained as a significant predictor of HCC (HR 1.904, P = .01). In vitro experiments revealed that compared with mock-transduced immortalized liver cells, CAV1-transduced cells showed significantly increased proliferation (P < .001) and colony formation in soft agar (P < .033). TMA with immunohistochemistry showed that tissues with CAV1 expression were more likely to develop HCC than tissues without CAV1 expression (P = .047). CAV1 expression predicts HCC development, making it a potential biomarker and target for preventive therapy.


Stress & the gut-brain axis: Regulation by the microbiome.
The importance of the gut-brain axis in regulating stress-related responses has long been appreciated. More recently, the microbiota has emerged as a key player in the control of this axis, especially during conditions of stress provoked by real or perceived homeostatic challenge. Diet is one of the most important modifying factors of the microbiota-gut-brain axis. The routes of communication between the microbiota and brain are slowly being unravelled, and include the vagus nerve, gut hormone signaling, the immune system, tryptophan metabolism, and microbial metabolites such as short chain fatty acids. The importance of the early life gut microbiota in shaping later health outcomes also is emerging. Results from preclinical studies indicate that alterations of the early microbial composition by way of antibiotic exposure, lack of breastfeeding, birth by Caesarean section, infection, stress exposure, and other environmental influences - coupled with the influence of host genetics - can result in long-term modulation of stress-related physiology and behaviour. The gut microbiota has been implicated in a variety of stress-related conditions including anxiety, depression and irritable bowel syndrome, although this is largely based on animal studies or correlative analysis in patient populations. Additional research in humans is sorely needed to reveal the relative impact and causal contribution of the microbiome to stress-related disorders. In this regard, the concept of psychobiotics is being developed and refined to encompass methods of targeting the microbiota in order to positively impact mental health outcomes. At the 2016 Neurobiology of Stress Workshop in Newport Beach, CA, a group of experts presented the symposium "The Microbiome: Development, Stress, and Disease". This report summarizes and builds upon some of the key concepts in that symposium within the context of how microbiota might influence the neurobiology of stress.


Granulicatella adiacens and Abiotrophia defectiva bacteraemia characterized by 16S rRNA gene sequencing.
Traditionally, the identification, epidemiology and spectrum of clinical diseases caused by Granulicatella adiacens and Abiotrophia defectiva are dependent upon their phenotypic characterization. During a 6-year period (July 1995-June 2001), seven and two alpha-haemolytic streptococci were identified as G. adiacens and A. defectiva, respectively, by 16S rRNA gene sequencing. Three patients with haematological malignancies and neutropenic fever had primary bacteraemia. Three patients with valvular problems or congenital heart disease had infective endocarditis. A patient with ischemic heart disease and cerebrovascular accident had infected aortic atheroma with dissection. A patient with recurrent pyogenic cholangitis had acute cholangitis and a patient with polypoid cystitis and benign prostatic hypertrophy had acute prostatitis. Four of the nine patients died, including all three with G. adiacens infective endocarditis or infected atheroma. For the seven G. adiacens isolates, the API 20 STREP system successfully identified one and five isolates as G. adiacens with >95 % and 80-90 % confidence, respectively, whereas the Vitek System (GPI) and ATB Expression system (ID32 STREP) successfully identified none and one isolate as G. adiacens. Of the two A. defectiva isolates, none of the three systems successfully identified either of them as A. defectiva. 16S rRNA gene sequencing is the technique of choice for identifying G. adiacens and A. defectiva, and early surgical intervention should be considered when G. adiacens endocarditis is diagnosed.


Coherent theta oscillations and reorganization of spike timing in the hippocampal- prefrontal network upon learning.
To study the interplay between hippocampus and medial prefrontal cortex (Pfc) and its importance for learning and memory consolidation, we measured the coherence in theta oscillations between these two structures in rats learning new rules on a Y maze. Coherence peaked at the choice point, most strongly after task rule acquisition. Simultaneously, Pfc pyramidal neurons reorganized their phase, concentrating at hippocampal theta trough, and synchronous cell assemblies emerged. This synchronous state may result from increased inhibition exerted by interneurons on pyramidal cells, as measured by cross-correlation, and could be modulated by dopamine: we found similar hippocampal-Pfc theta coherence increases and neuronal phase shifts following local administration of dopamine in Pfc of anesthetized rats. Pfc cell assemblies emerging during high coherence were preferentially replayed during subsequent sleep, concurrent with hippocampal sharp waves. Thus, hippocampal/prefrontal coherence could lead to synchronization of reward predicting activity in prefrontal networks, tagging it for subsequent memory consolidation.


Rokubacteria: Genomic Giants among the Uncultured Bacterial Phyla.
Recent advances in single-cell genomic and metagenomic techniques have facilitated the discovery of numerous previously unknown, deep branches of the tree of life that lack cultured representatives. Many of these candidate phyla are composed of microorganisms with minimalistic, streamlined genomes lacking some core metabolic pathways, which may contribute to their resistance to growth in pure culture. Here we analyzed single-cell genomes and metagenome bins to show that the "Candidate phylum Rokubacteria," formerly known as SPAM, represents an interesting exception, by having large genomes (6-8 Mbps), high GC content (66-71%), and the potential for a versatile, mixotrophic metabolism. We also observed an unusually high genomic heterogeneity among individual Rokubacteria cells in the studied samples. These features may have contributed to the limited recovery of sequences of this candidate phylum in prior cultivation and metagenomic studies. Our analyses suggest that Rokubacteria are distributed globally in diverse terrestrial ecosystems, including soils, the rhizosphere, volcanic mud, oil wells, aquifers, and the deep subsurface, with no reports from marine environments to date.


Protein Interactomic Analysis of SAPKs and ABA-Inducible bZIPs Revealed Key Roles of SAPK10 in Rice Flowering.
As core components of ABA signaling pathway, SnRK2s (Sucrose nonfermenting1⁻Related protein Kinase 2) bind to and phosphorylate AREB/ABF (ABA responsive element binding protein/ABRE-binding factor) transcriptional factors, particularly bZIPs (basic region-leucine zipper), to participate in various biological processes, including flowering. Rice contains 10 SnRK2 members denoted as SAPK1-10 (Stress-Activated Protein Kinase) and dozens of bZIPs. However, which of the SAPKs and bZIPs pair and involve in ABA signaling remains largely unknown. In this study, we carried out a systematical protein-protein interactomic analysis of 10 SAPKs and 9 ABA-inducible bZIPs using yeast-two-hybrid technique, and identified 14 positive interactions. The reliability of Y2H work was verified by in vitro pull-down assay of the key flowering regulator bZIP77 with SAPK9 and SAPK10, respectively. Moreover, SAPK10 could phosphorylate bZIP77 in vitro. Over-expression of SAPK10 resulted in earlier flowering time, at least partially through regulating the FAC-MADS15 pathway. Conclusively, our results provided an overall view of the SAPK-bZIP interactions, and shed novel lights on the mechanisms of ABA-regulated rice flowering.


Sociopathy as a human process. A viewpoint.
Case histories of narcotic addicts who also were imprisoned for felony were selected to illustrate some underlying dynamics of Cleckley's so-called psychopath and some principles useful in their management. Often in outpatient settings, such individuals seem to be without anxiety, unable to experience depression, and without motivation for recovery; but in inpatient settings, such deficits appear illusory. Once such chronically sociopathic individuals are prevented from 'running' their resemblance to individuals with severe but thoroughly 'human' and comprehensible personality disorders becomes evident. In treatment, external controls are important. It is vital both to appreciate the contagion of the psychopath's invisible anxiety and to provide such individuals with alternative defenses with which to mitagate their depression. Finally sociopaths must be realistically, but not puntively, confronted with consequences of their behavior.


Effect of Diet Supplementation with the Mycotoxin Binder Montmorillonite on the Bioavailability of Vitamins in Dairy Cows.
The objective of this study was to determine the effect of the mycotoxin binder montmorillonite (MMT) supplemented in the diet of dairy cows on the bioavailability of vitamins A, D, E, B1 and B6. Six multiparous Holstein-Friesian cows were used in a crossover design with two periods. Treatments were a control diet with or without MMT. Vitamins were infused individually into the abomasum through the ruminal cannula. Blood samples were collected from the jugular vein at 0, 1, 2, 3, 4, 6, 9, 12, 24 and 48 h after the administration of each vitamin. Results showed that vitamin A reached maximal concentration (Tmax) at 5.3 h after dosing, the maximal concentration (Cmax) was 1.2 times higher than the basal concentration (Cbasal), and the area under the curve (AUC) was 739 arbitrary units. Vitamin B6 reached the Tmax at 13 h after dosing, the Cmax was 1.4 times higher than the Cbasal, and the AUC was 222 arbitrary units. No differences were observed in Cbasal, Tmax, Cmax and AUC of vitamin A and B6 between control vs. MMT-supplemented cows. Plasma concentrations of vitamins D, E and B1 had no concentration peaks, and were not affected by MMT addition. The lack of a response suggests that their plasma concentration may be tightly regulated. Results of this study do not show evidence that MMT affects the bioavailability of vitamins A and B6 in vivo.


Leader peptide and a membrane protein scaffold guide the biosynthesis of the tricyclic peptide microviridin.
Microviridins are unique protease inhibitors from bloom-forming cyanobacteria that have both ecological and pharmacological relevance. Their peptide backbones are produced ribosomally, and ATP grasp ligases introduce ω-ester and ω-amide bonds to yield rare cage-like structures. Bioinformatic analysis of the microviridin biosynthesis gene cluster suggests a novel type of processing machinery, which could rationalize the challenging in vivo/in vitro reconstitution of the pathway. In this work, we report the establishment of a minimal expression system for microviridins. Through bioinformatics and mutational analysis of the MdnA leader peptide we identified and characterized a strictly conserved binding motif that is specific for microviridin ligases. Furthermore, we showed that the ABC transporter MdnE is crucial for cyclization and processing of microviridins and demonstrated that MdnE is essential for stability of the microviridin biosynthesis complex.


Kabuki syndrome: international consensus diagnostic criteria.
Kabuki syndrome (KS) is a clinically recognisable syndrome in which 70% of patients have a pathogenic variant in KMT2D or KDM6A. Understanding the function of these genes opens the door to targeted therapies. The purpose of this report is to propose diagnostic criteria for KS, particularly when molecular genetic testing is equivocal. An international group of experts created consensus diagnostic criteria for KS. Systematic PubMed searches returned 70 peer-reviewed publications in which at least one individual with molecularly confirmed KS was reported. The clinical features of individuals with known mutations were reviewed. The authors propose that a definitive diagnosis can be made in an individual of any age with a history of infantile hypotonia, developmental delay and/or intellectual disability, and one or both of the following major criteria: (1) a pathogenic or likely pathogenic variant in KMT2D or KDM6A; and (2) typical dysmorphic features (defined below) at some point of life. Typical dysmorphic features include long palpebral fissures with eversion of the lateral third of the lower eyelid and two or more of the following: (1) arched and broad eyebrows with the lateral third displaying notching or sparseness; (2) short columella with depressed nasal tip; (3) large, prominent or cupped ears; and (4) persistent fingertip pads. Further criteria for a probable and possible diagnosis, including a table of suggestive clinical features, are presented. As targeted therapies for KS are being developed, it is important to be able to make the correct diagnosis, either with or without molecular genetic confirmation.


The incidence of trisomy 8 as a sole chromosomal aberration in myeloid malignancies varies in relation to gender, age, prior iatrogenic genotoxic exposure, and morphology.
Although trisomy 8 as a sole change is one of the most common chromosomal abnormalities in myeloid malignancies, it is largely unknown if the incidence of this aberration is influenced by other factors of clinical importance. In the present study, the frequencies of isolated +8 in relation to gender, age, previous treatment with chemo- or radiotherapy, and morphologic subtype were ascertained in published, as well as in our own unpublished, cases of acute myeloid leukemia (AML; n=4,246), myelodysplastic syndromes (MDS; n=1,817), and chronic myeloproliferative disorders (MPD; n=530). The frequencies of +8 were higher in MDS and MPD than in AML (7.5% vs. 5.6%; P<0.01) and varied among the morphologic subtypes of AML and MDS (P<0.001 and P<0.05, respectively). Trisomy 8 was more common in women than in men with MPD (11% vs. 5.1%; P<0.05). Furthermore, the frequencies of +8 were higher in de novo AML and MDS than in treatment-related cases (6.0% vs. 2.8%; P<0.01 and 8.6% vs. 1.5%; P<0.001, respectively). The incidence also varied significantly with age in AML (P<0.001), being more common in elderly patients. Although the causes for this frequency heterogeneity remain to be elucidated, possible explanations may include different environmental exposures affecting the origin of +8 in AML, MDS, and MPD and the presence of different underlying cryptic primary aberrations.


An analysis of research quality underlying IDSA clinical practice guidelines: a cross-sectional study.
As a result of new developments in medicine, the need for evidence-based clinical practice guidelines (CPG) is of utmost importance. However, studies have shown that many medical societies are using low quality research to develop CPGs. To evaluate the quality of research underlying the CPGs issued by the Infectious Diseases Society of America (IDSA). We examined 29 CPGs issued between January 1, 2012 and December 31, 2019 and classified each by research quality according to levels reported by the CPG authors and previously specified by the IDSA: Levels I through III, corresponding to high, moderate, and low quality of evidence, respectively. Each ranking was cross-checked with a second researcher to improve inter-rater reliability. To analyze evolution of research quality over time, three updated CPGs were randomly selected and compared to their original versions. Chi-square analysis was then performed to determine statistical significance. We evaluated the quality of research for 2,920 recommendations within the 29 CPGs that met our criteria and found that 418 (14%) were developed using high-quality (Level I) research from randomized, controlled trials. Of the remaining recommendations, 928 (32%) were based on moderate quality research (observational studies) and 1574 (54%) on low quality research (expert opinion). A Pearson chi-squared analysis indicated no-statistically significant difference between original guidelines or their subsequent updates for Clostridium difficile (χ2=0.323; n=85; degrees of freedom [df]=2; p=0.851), candidiasis (χ2=4.133; n=195; df=2; p=0.127), or coccidiomycosis (χ2=0.531; n=95; df=1; p=0.466). The proportion of high-quality research underlying guideline recommendations is remarkably low, indicating that moderate and low quality evidence is still influencing infectious disease guidelines despite IDSA standards. Moreover, the quality of research has not significantly changed over time. IDSA CPGs are a formidable source of information for clinicians, but an increased number of quality studies should be utilized to further guide CPG development.


Inflammation and chronic oxidative stress in radiation-induced late normal tissue injury: therapeutic implications.
The threat of radiation-induced late normal tissue injury limits the dose of radiation that can be delivered safely to cancer patients presenting with solid tumors. Tissue dysfunction and failure, associated with atrophy, fibrosis and/or necrosis, as well as vascular injury, have been reported in late responding normal tissues, including the central nervous system, gut, kidney, liver, lung, and skin. The precise mechanisms involved in the pathogenesis of radiation-induced late normal tissue injury have not been fully elucidated. It has been proposed recently that the radiation-induced late effects are caused, in part, by chronic oxidative stress and inflammation. Increased production of reactive oxygen species, which leads to lipid peroxidation, oxidation of DNA and proteins, as well as activation of pro-inflammatory factors has been observed in vitro and in vivo. In this review, we will present direct and indirect evidence to support this hypothesis. To improve the long-term survival and quality of life for radiotherapy patients, new approaches have been examined in preclinical models for their efficacy in preventing or mitigating the radiation-induced chronic normal tissue injury. We and others have tested drugs that can either attenuate inflammation or reduce chronic oxidative stress in animal models of late radiation-induced normal tissue injury. The effectiveness of renin-angiotensin system blockers, peroxisome proliferator-activated receptor (PPAR) gamma agonists, and antioxidants/antioxidant enzymes in preventing or mitigating the severity of radiation-induced late effects indicates that radiation-induced chronic injury can be prevented and/or treated. This provides a rationale for the design and development of anti-inflammatory-based interventional approaches for the treatment of radiation-induced late normal tissue injury.


Possible impact of stimulus duration on seizure threshold in ECT.
Seizure thresholds were measured in patients treated with right unilateral electroconvulsive therapy (ECT) using 0.9 amp, 0.5 ms constant current pulses at 30 Hz for 0.93-2.8 s. Using these parameters, only two patients failed to seize at stimulus charges < or = 75.6 mC. Three patients were excluded from further analysis because they experienced generalized motor or electrographic seizures lasting less than 25 s and 30 s, respectively. The remaining 52 patients had a mean seizure threshold of 48.9 +/- 15.3 mC. Seizure threshold was correlated with age, but not sex. These values for seizure threshold are significantly lower than those reported previously using longer pulse-briefer train duration stimuli, suggesting that characteristics of the electrical stimulus influence seizure threshold. These findings have implications for electrical dosing in right unilateral ECT.


A reality check for climate change experiments: Do they reflect the real world?
Experiments are widely used in ecology, particularly for assessing global change impacts on ecosystem function. However, results from experiments often are inconsistent with observations made under natural conditions, suggesting the need for rigorous comparisons of experimental and observational studies. We conducted such a "reality check" for a grassland ecosystem by compiling results from nine independently conducted climate change experiments. Each experiment manipulated growing season precipitation (GSP) and measured responses in aboveground net primary production (ANPP). We compared results from experiments with long-term (33-yr) annual precipitation and ANPP records to ask if collectively (n = 44 experiment-years) experiments yielded estimates of ANPP, rain-use efficiency (RUE, grams per square meter ANPP per mm precipitation), and the relationship between GSP and ANPP comparable to observations. We found that mean ANPP and RUE from experiments did not deviate from observations. Experiments and observational data also yielded similar functional relationships between ANPP and GSP, but only within the range of historically observed GSP. Fewer experiments imposed extreme levels of GSP (outside the observed 33-yr record), but when these were included, they altered the GSP-ANPP relationship. This result underscores the need for more experiments imposing extreme precipitation levels to resolve how forecast changes in climate regimes will affect ecosystem function in the future.


The focal adhesion kinase Pyk2 links Ca2+ signalling to Src family kinase activation and protein tyrosine phosphorylation in thrombin-stimulated platelets.
In blood platelets, stimulation of G protein-coupled receptors (GPCRs) by thrombin triggers the activation of Src family kinases (SFKs), resulting in the tyrosine-phosphorylation of multiple substrates, but the mechanism underlying this process is still poorly understood. In the present study, we show that the time-dependent protein-tyrosine phosphorylation triggered by thrombin in human or murine platelets was totally suppressed only upon concomitant chelation of intracellular Ca(2+) and inhibition of SFKs. Thrombin-induced activation of SFKs was regulated by intracellular Ca(2+) and accordingly the Ca(2+) ionophore A23187 was sufficient to stimulate SFKs. A23187 also triggered the phosphorylation and activation of the Ca(2+)-dependent focal adhesion kinase Pyk2 and Pyk2 activation by thrombin was Ca(2+)-dependent. Stimulation of SFKs by thrombin or A23187 was strongly reduced in platelets from Pyk2 knockout (KO) mice, as was the overall pattern of protein-tyrosine phosphorylation. By immunoprecipitation experiments, we demonstrate that Lyn and Fyn, but not Src, were activated by Pyk2. Inhibition of SFKs by PP2 also reduced the phosphorylation of Pyk2 in thrombin or A23187-stimulated platelets. Analysis of KO mice demonstrated that Fyn, but not Lyn, was required for complete Pyk2 phosphorylation by thrombin. Finally, PP2 reduced aggregation of murine platelets to a level comparable to that of Pyk2-deficient platelets, but did not have further effects in the absence of Pyk2. These results indicate that in thrombin-stimulated platelets, stimulation of Pyk2 by intracellular Ca(2+) initiates SFK activation, establishing a positive loop that reinforces the Pyk2/SFK axis and allows the subsequent massive tyrosine phosphorylation of multiple substrates required for platelet aggregation.


Dermal exposure assessment in occupational epidemiologic research.
Recognition of the importance of skin exposure in industrial settings has steadily increased over the last few decades. Unfortunately, the growing attention to dermal exposure in industrial hygiene has often not been reflected in the field of occupational epidemiology. An extensive literature survey was conducted to identify dermal exposure assessment methods that have been applied in epidemiologic studies. Subsequently, methodologies are postulated that could be applied to epidemiologic research. Attention is given to intensity, frequency, and duration of exposure, the exposed surface area, and personal, temporal and spatial variability in dermal exposure and uptake. It is anticipated that, in the near future, dermal exposure assessment in epidemiologic research will be based generally on expert judgment and to some degree on process-specific exposure models. Field studies collecting quantitative dermal exposure data and statistical modeling to identify exposure determinants will, however, be imperative if progress is to be made in the field of dermal exposure assessment for epidemiologic purposes.


Placental miR-1301 is dysregulated in early-onset preeclampsia and inversely correlated with maternal circulating leptin.
miRNAs are small non-coding RNAs important for the regulation of mRNA in many organs including placenta. Adipokines and specifically leptin are known to be dysregulated in preeclampsia, but little is known regarding their regulation by miRNAs during pregnancy. We performed high-throughput sequencing of small RNAs in placenta from 72 well-defined patients: 23 early-onset preeclampsia (PE), 26 late-onset PE and 23 controls. The regulation of some miRNAs was confirmed on qRT-PCR. Maternal circulating levels and placental mRNA of leptin, resistin and adiponectin were measured using Bio-Plex and qRT-PCR. We found that miR-1301, miR-223 and miR-224 expression was downregulated in early-onset PE, but not in late-onset PE, compared to controls. In silico analysis predicted the leptin gene (LEP) to be a target for all three miRNAs. Indeed, we found significant correlation between maternal circulating levels of leptin and placental LEP expression. In addition, we found a significant inverse correlation between maternal circulating leptin/placental LEP expression and placental miR-1301 expression levels. Interestingly, placental expression of miR-1301 was also correlated with newborn weight percentile and inversely correlated with both maternal systolic and diastolic blood pressure prior to delivery. Our results confirm that placenta is a major site of LEP expression during pregnancy. It further suggests that miR-1301 could be involved in the regulation of leptin during pregnancy and may play a role in early-onset PE. miR-1301 is dysregulated in early-onset preeclampsia and could possibly play a role in the regulation of leptin during pregnancy.


Waging war on physical inactivity: using modern molecular ammunition against an ancient enemy.
A hypothesis is presented based on a coalescence of anthropological estimations of Homo sapiens' phenotypes in the Late Paleolithic era 10,000 years ago, with Darwinian natural selection synergized with Neel's idea of the so-called thrifty gene. It is proposed that humans inherited genes that were evolved to support a physically active lifestyle. It is further postulated that physical inactivity in sedentary societies directly contributes to multiple chronic health disorders. Therefore, it is imperative to identify the underlying genetic and cellular/biochemical bases of why sedentary living produces chronic health conditions. This will allow society to improve its ability to effect beneficial lifestyle changes and hence improve the overall quality of living. To win the war against physical inactivity and the myriad of chronic health conditions produced because of physical inactivity, a multifactorial approach is needed, which includes successful preventive medicine, drug development, optimal target selection, and efficacious clinical therapy. All of these approaches require a thorough understanding of fundamental biology and how the dysregulated molecular circuitry caused by physical inactivity produces clinically overt disease. The purpose of this review is to summarize the vast armamentarium at our disposal in the form of the extensive scientific basis underlying how physical inactivity affects at least 20 of the most deadly chronic disorders. We hope that this information will provide readers with a starting point for developing additional strategies of their own in the ongoing war against inactivity-induced chronic health conditions.


Pinpointing beta adrenergic receptor in ageing pathophysiology: victim or executioner? Evidence from crime scenes.
G protein-coupled receptors (GPCRs) play a key role in cellular communication, allowing human cells to sense external cues or to talk each other through hormones or neurotransmitters. Research in this field has been recently awarded with the Nobel Prize in chemistry to Robert J. Lefkowitz and Brian K. Kobilka, for their pioneering work on beta adrenergic receptors (βARs), a prototype GPCR. Such receptors, and β2AR in particular, which is extensively distributed throughout the body, are involved in a number of pathophysiological processes. Moreover, a large amount of studies has demonstrated their participation in ageing process. Reciprocally, age-related changes in regulation of receptor responses have been observed in numerous tissues and include modifications of βAR responses. Impaired sympathetic nervous system function has been indeed evoked as at least a partial explanation for several modifications that occur with ageing. This article represents an updated presentation of the current knowledge in the field, summarizing in a systematic way the major findings of research on ageing in several organs and tissues (crime scenes) expressing βARs: heart, vessels, skeletal muscle, respiratory system, brain, immune system, pancreatic islets, liver, kidney and bone.


3D Printed Hydrogels with Aligned Microchannels to Guide Neural Stem Cell Migration.
Following traumatic or ischemic brain injury, rapid cell death and extracellular matrix degradation lead to the formation of a cavity at the brain lesion site, which is responsible for prolonged neurological deficits and permanent disability. Transplantation of neural stem/progenitor cells (NSCs) represents a promising strategy for reconstructing the lesion cavity and promoting tissue regeneration. In particular, the promotion of neuronal migration, organization, and integration of transplanted NSCs is critical to the success of stem cell-based therapy. This is particularly important for the cerebral cortex, the most common area involved in brain injuries, because the highly organized structure of the cerebral cortex is essential to its function. Biomaterials-based strategies show some promise for conditioning the lesion site microenvironment to support transplanted stem cells, but the progress in demonstrating organized cell engraftment and integration into the brain is very limited. An effective approach to sufficiently address these challenges has not yet been developed. Here, we have implemented a digital light-processing-based 3D printer and printed hydrogel scaffolds with a designed shape, uniaxially aligned microchannels, and tunable mechanical properties. We demonstrated the capacity to achieve high shape precision to the lesion site with brain tissue-matching mechanical properties. We also established spatial control of bioactive molecule distribution within 3D printed hydrogel scaffolds. These printed hydrogel scaffolds have shown high neuro-compatibility with aligned neuronal outgrowth along with the microchannels. This study will provide a biomaterial-based approach that can serve as a protective and guidance vehicle for transplanted NSC organization and integration for brain tissue regeneration after injuries.


Hospitalizations for Endocarditis and Associated Health Care Costs Among Persons with Diagnosed Drug Dependence - North Carolina, 2010-2015.
Opioid dependence and overdose have increased to epidemic levels in the United States. The 2014 National Survey on Drug Use and Health estimated that 4.3 million persons were nonmedical users of prescription pain relievers (1). These users are 40 times more likely than the general population to use heroin or other injection drugs (2). Furthermore, CDC estimated a near quadrupling of heroin-related overdose deaths during 2002-2014 (3). Although overdose contributes most to drug-associated mortality, infectious complications of intravenous drug use constitute a major cause of morbidity leading to hospitalization (4). In addition to infections from hepatitis C virus (HCV) and human immunodeficiency virus (HIV), injecting drug users are at increased risk for acquiring invasive bacterial infections, including endocarditis (5,6). Evidence that hospitalizations for endocarditis are increasing in association with the current opioid epidemic exists (7-9). To examine trends in hospitalizations for endocarditis among persons in North Carolina with drug dependence during 2010-2015, data from the North Carolina Hospital Discharge database were analyzed. The incidence of hospital discharge diagnoses for drug dependence combined with endocarditis increased more than twelvefold from 0.2 to 2.7 per 100,000 persons per year over this 6-year period. Correspondingly, hospital costs for these patients increased eighteenfold, from $1.1 million in 2010 to $22.2 million in 2015. To reduce the risk for morbidity and mortality related to opioid-associated endocarditis, public health programs and health care systems should consider collaborating to implement syringe service programs, harm reduction strategies, and opioid treatment programs.


The Impact of Different Inoculation Schemes on the Microbiota, Physicochemical and Sensory Characteristics of Greek Kopanisti Cheese throughout Production and Ripening.
Kopanisti is a Greek PDO cheese, which is traditionally produced by the addition of an amount of over-mature Kopanisti, called Mana Kopanisti, to initiate cheese ripening. The aim of this study was the production of four types of Kopanisti cheese (A-D) using pasteurized cow milk, and a combination of the following starters/adjuncts in order to test their ability to be used in Kopanisti cheese production: A: Lactococcus lactis subsp. lactis and Lacticaseibacillus paracasei, B: L. lactis and Lc. paracasei/Mana Kopanisti, C: L. lactis and Lc. paracasei/Ligilactobacillus acidipiscis and Loigolactobacillus rennini, D: Lig. acidipiscis and Loig. rennini. Throughout production and ripening, classical microbiological, metataxonomics and physicochemical analyses were employed, while the final products (Day 35) were subjected to sensory analysis as well. Most interestingly, beta-diversity analysis of the metataxonomics data revealed the clusters constructed among the Kopanisti types based on the different inoculation schemes. On day 35, Kopanisti A-C types clustered together due to their similar 16S microbiota, while Kopanisti D was highly differentiated. On the contrary, ITS data clustered Kopanisti B and C together, while Kopanisti A and D were grouped seperately. Finally, based on the sensory evaluation, Kopanisti C appeared to have the most suitable bacteria cocktail for the Kopanisti cheese production. Therefore, not only were the conventional starters used, but also the Lig. acidipiscis and Loig. rennini strains could be used in a standardized Kopanisti cheese production that could lead to final products of high quality and safety.


The role of antimicrobial peptides at the ocular surface.
Antimicrobial peptides (AMPs) such as defensins and cathelicidins are small peptides with broad-spectrum activity against bacteria, fungi and viruses. In addition, several AMPs modulate mammalian cell behaviours including migration, proliferation and cytokine production. This review describes findings from recent studies showing the presence of various AMPs at the human ocular surface and discusses their mechanism of antimicrobial action and potential non-microbicidal roles. Corneal and conjunctival epithelial cells produce beta-defensins and the cathelicidin LL-37, whereas neutrophils, infiltrating in response to a specific stimulus, supply additional LL-37 as well as alpha-defensins. In vitro studies suggest that LL-37 and human beta-defensin-3 are the most likely to have significant independent antimicrobial activity, while other AMPs may act synergistically to help protect the ocular surface from invading pathogens. Current evidence also supports a role for some AMPs in modulating wound healing responses. Although yet to be brought to fruition, AMPs hold significant potential as therapeutic agents for the prophylaxis and treatment of infection, promotion of wound healing and immune modulation.


Effects of dietary cholesterol and simvastatin on cholesterol synthesis in Smith-Lemli-Opitz syndrome.
Deficient cholesterol and/or excessive 7-dehydrocholesterol (7-DHC) may be responsible for the pathology of Smith-Lemli-Opitz syndrome (SLOS). Both high-cholesterol diets given to ameliorate cholesterol deficiency while decreasing 7-DHC and cholesterol-enriched diets plus simvastatin to further decrease sterol synthesis have been used as potential therapies. However, the effect of dietary cholesterol and simvastatin on cholesterol synthesis in SLOS has not been reported. Twelve subjects with SLOS enrolled in the study: Nine had received a high cholesterol diet (HI) for 3 y and three were studied after 4 wk on a low cholesterol diet (LO). Cholesterol fractional synthesis rate (FSR) was measured after oral administration of deuterium oxide, using gas chromatography isotope ratio mass spectrometry. FSR was lower in HI compared with LO (HI: 1.46 +/- 0.62%/d; LO: 4.77 +/- 0.95%/d; p < 0.001). Three HI subjects were retested after 0.8 y taking simvastatin (HI + ST). Simvastatin tended to reduce FSR and significantly decreased (p < 0.01) plasma 7-DHC compared with cholesterol supplementation alone. The study demonstrates the utility of the deuterium incorporation method to understand the effect of therapeutic interventions in SLOS. The data suggest that dietary cholesterol supplementation reduces cholesterol synthesis in SLOS and further support the rationale for the combined treatment of SLOS with a cholesterol-enriched diet and simvastatin.


Direct and indirect measurements of enhanced phenolic bioavailability from litchi pericarp procyanidins by Lactobacillus casei-01.
Litchi pericarp procyanidins (LPP) are dietary supplements with high antioxidant activity, but poor oral bioavailability and efficacy. Lactobacillus casei (L. casei-01) can transform flavan-3-ols from litchi pericarp and increase their antioxidant ability; thus, L. casei-01 with LPP was administered to rats for four and eight weeks to study the effect of such a combination on metabolic parameters and on phase II metabolism and detoxification pathways in the liver as an indirect measure for phenolic bioavailability. Our data indicated that the T-AOC of the plasma, the liver GSH-Px and GSH-ST activity, and the expression of UGT and SULT isoforms in the liver of the rats were all enhanced after the eight-week administration compared with those of the control. However, at 1 h after administration the concentration of (-)-epicatechin in the combined system was lower than that obtained after the ingestion of LPP alone, suggesting that L. casei-01 enhances the bioavailability of phenolics from LPP by modulating the transformation to other compounds but not by increasing its absorption in the native form.


Preconception care between pregnancies: the content of internatal care.
For more than two decades, prenatal care has been a cornerstone of our nation's strategy for improving pregnancy outcomes. In recent years, however, a growing recognition of the limits of prenatal care and the importance of maternal health before pregnancy has drawn increasing attention to preconception and internatal care. Internatal care refers to a package of healthcare and ancillary services provided to a woman and her family from the birth of one child to the birth of her next child. For healthy mothers, internatal care offers an opportunity for wellness promotion between pregnancies. For high-risk mothers, internatal care provides strategies for risk reduction before their next pregnancy. In this paper we begin to define the contents of internatal care. The core components of internatal care consist of risk assessment, health promotion, clinical and psychosocial interventions. We identified several priority areas, such as FINDS (family violence, infections, nutrition, depression, and stress) for risk assessment or BBEEFF (breastfeeding, back-to-sleep, exercise, exposures, family planning and folate) for health promotion. Women with chronic health conditions such as hypertension, diabetes, or weight problems should receive on-going care per clinical guidelines for their evaluation, treatment, and follow-up during the internatal period. For women with prior adverse outcomes such as preterm delivery, we propose an internatal care model based on known etiologic pathways, with the goal of preventing recurrence by addressing these biobehavioral pathways prior to the next pregnancy. We suggest enhancing service integration for women and families, including possibly care coordination and home visitation for selected high-risk women. The primary aim of this paper is to start a dialogue on the content of internatal care.


On Detectable and Meaningful Speech-Intelligibility Benefits.
The most important parameter that affects the ability to hear and understand speech in the presence of background noise is the signal-to-noise ratio (SNR). Despite decades of research in speech intelligibility, it is not currently known how much improvement in SNR is needed to provide a meaningful benefit to someone. We propose that the underlying psychophysical basis to a meaningful benefit should be the just noticeable difference (JND) for SNR. The SNR JND was measured in a series of experiments using both adaptive and fixed-level procedures across participants of varying hearing ability. The results showed an average SNR JND of approximately 3 dB for sentences in same-spectrum noise. The role of the stimulus and link to intelligibility was examined by measuring speech-intelligibility psychometric functions and comparing the intelligibility JND estimated from those functions with measured SNR JNDs. Several experiments were then conducted to establish a just meaningful difference (JMD) for SNR. SNR changes that could induce intervention-seeking behaviour for an individual were measured with subjective scaling and report, using the same stimuli as the SNR JND experiment as pre- and post-benefit examples. The results across different rating and willingness-to-change tasks showed that the mean ratings increased near linearly with a change in SNR, but a change of at least 6 dB was necessary to reliably motivate participants to seek intervention. The magnitude of the JNDs and JMDs for speech-intelligibility benefits measured here suggest a gap between what is achievable and what is meaningful.


Touchdown General Primer (GP5+/GP6+) PCR and optimized sample DNA concentration support the sensitive detection of human papillomavirus.
The GP5+/GP6+ PCR assay is a well-established HPV detection technique. This study has examined the effects of incorporating 'hot start' and 'touchdown' steps into the protocol. In addition, dTTP was substituted with dUTP to permit contamination control measures against carry-over PCR product. Firstly, HPV-16 was amplified from SiHa cell DNA (0.1 ng-100 ng) diluted in a background of C-33A DNA (100 ng-2 microg). Secondly, the detection of small quantities (15ag-1.5pg) of HPV recombinant plasmids (types 16, 31, 33, 45, 51, 52, and 56) diluted in C-33A DNA was investigated. Thirdly, clinical sample DNA extracts (cervical smears, formalin-fixed vaginal lesions and breast tumors) were tested for HPV. Six different PCR protocols were assessed. HPV was detected by gel electrophoresis, and by Southern and dot blot hybridization. HPV detection sensitivity was dependent on the total amount of DNA in a PCR. Touchdown protocols supported HPV-16 detection from 1 ng or 0.5 ng SiHa cell DNA in a background of 2 microg or 1 microg C-33A DNA respectively, and from 0.1 ng of SiHa cell DNA (approximately 28 copies HPV-16) in 500 ng or 100 ng background DNA. Under standard GP5+/GP6+ annealing conditions, HPV-16 went undetected when the DNA content of a PCR was 2 microg or 1 microg, and with 500 ng C-33A DNA the sensitivity limit was 1 ng SiHa cell DNA. HPV recombinant plasmids were each detected with high (albeit varying) sensitivity by a touchdown protocol. HPV-31 was better amplified under standard annealing conditions (1.5fg in 100 ng background DNA) than by a touchdown approach (15fg detection limit). HPV-52 was not amplified by the standard protocol at the dilutions tested. Seventeen different HPV types were demonstrated in 47/65 (72%) abnormal cytology samples recorded as HPV negative by standard GP5+/GP6+ conditions. Twenty-one different HPV types were recorded in 111/114 (97%) vaginal lesions. Multiple infections were also detectable using a touchdown approach. Of 26 breast tumors, 5 (19%) tested HPV positive by the standard assay and 15/26 (58%) using a touchdown protocol. Touchdown modification of the GP5+/GP6+ PCR assay enables the detection of HPV undetected under regular assay conditions. The use of standardized DNA quantities in a PCR rather than standard sample volumes containing arbitrary amounts of DNA is supported. A touchdown approach may be beneficial as an analytical test for the re-evaluation of (apparently) HPV negative abnormal cervical cytological or histological samples, and for investigating the association of HPV with disease conditions at diverse organ sites. The clinical utility of a touchdown approach for HPV detection requires further investigation as increased assay analytical sensitivity may not necessarily equate with improved clinical sensitivity or specificity.


The response of infants with bronchiolitis to the proteins of respiratory syncytial virus.
Acute phase sera were collected from 28 infants hospitalized with bronchiolitis due to respiratory syncytial (RS) virus and convalescent sera were collected from 24 of them. The sera were assayed for neutralizing antibodies by plaque inhibition, for antibodies to the viral proteins by Western blot against partially purified RS virus, and for their ability to inhibit attachment and fusion. Among the 28 acute phase sera, 27 had antibody to the attachment glycoprotein (G), 16 had antibody to the fusion glycoprotein (F), but none had antibody to the matrix protein (VPM). Both the geometric mean anti-G titre, and the geometric mean anti-F titre correlated with the 50% neutralizing dose (ND50) titre in the acute phase serum. Among the 24 convalescent sera, only four exhibited an increase in neutralizing antibody titre. The response to G appeared to be related to the acute phase ND50 titre. Of 17 infants with acute phase titres of less than 100 ND50/ml, 10 responded to G while there was no response to this protein in seven infants with acute phase titres greater than 100 ND50/ml. While only one infant responded to F, 18 responded to the phosphorylated nucleocapsid protein, VP32, and none responded to VPM. The ability of the acute phase sera to inhibit virus attachment to HeLa cells and to inhibit fusion correlated with the anti-G titre and the anti-F titre, respectively. However, there was no correlation between the inhibition of fusion and the anti-F titre in the convalescent sera, almost all of which inhibited fusion. These results suggest that the infected infants were responding to RS virus, but that their response to the viral proteins was either masked or slowed by residual maternal antibody. The inability to detect VPM in the acute and convalescent phase sera, as well as in 20 paired maternal and cord sera at a 1:50 dilution suggested that VPM, although it is one of the most prevalent viral proteins in both the virion and the infected cell, may be poorly antigenic in humans.


A case report of intrahepatic bile duct confluence anomalies in VACTERL syndrome.
The clinical manifestations of VACTERL association include vertebral anomalies, anal atresia, congenital heart diseases, tracheoesophageal fistula, renal dysplasia, and limb abnormalities. The association of intrahepatic anomalies and VACTERL syndrome is a rare coincidence. VACTER syndrome and intrahepatic bile drainage anomalies might be genetically related. A 12-year-old girl presented with episodic colicky abdominal pain, nausea, and vomiting for several years. The individual episodes resolved spontaneously within a few days. She had a history of VACTERL syndrome, including a butterfly shape of the L3 vertebra, anal atresia, and an atrial septal defect. On laboratory findings, abnormal liver function tests included elevated total bilirubin, alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyltransferase. There was no significant abnormal finding in hepatobiliary system sonography except mild gallbladder wall thickening. We performed magnetic resonance cholangiopancreatography and demonstrated an abnormal intrahepatic bile duct confluence, which showed 3 bile ducts draining directly into the neck of the gallbladder. Her symptoms related to bile reflux during gallbladder contraction. Cholecystectomy with choledochojejunostomy was undertaken because segments of the bile drainage were intertwined. After surgery, her symptoms decreased, but abdominal discomfort remained due to uncorrected left intrahepatic anomalies. Although hepatobiliary anomalies are not included in VACTERL association diagnostic criteria, detailed hepatobiliary work up is needed when gastrointestinal symptoms are present in VACTERL association patients.


Neurogenic aspects of stress urinary incontinence.
Vaginal childbirth is a significant risk factor for stress urinary incontinence (SUI). Women with SUI demonstrate dysfunction of the pelvic floor and pudendal nerve. Animal models of SUI have been developed to investigate its pathophysiology and for preclinical testing of potential treatments. Vaginal distension, a method of simulating childbirth injury in animals, produces a reliable decrease in leak point pressure (LPP), a measure of urethral resistance to leakage and quantification of SUI severity in animals. In addition to ischemia and direct tissue damage, vaginal distension causes denervation of the external urethral sphincter (EUS). Pudendal nerve crush produces a similar decrease in LPP, whereas combined PNC and vaginal distension injury delays recovery of LPP compared with either single injury alone. Neurophysiologic studies have elucidated the results of each injury and their combination on pudendal nerve and EUS function. Urethrolysis, electrocautery, and pudendal nerve transection produce more durable functional impairment via both structural damage and denervation. Pubourethral ligament injury eliminates the structural support of the urethra, but its neurologic effects are unknown. Animal models demonstrate a complex interplay between tissue damage and pudendal nerve dysfunction, and provide insight into the importance of neuroregeneration in the recovery of continence.


Proteomic characterization of acyclovir-induced nephrotoxicity in a mouse model.
Acyclovir (ACV) is an effective and widely used antiviral agent. However, its clinical application is limited by severe nephrotoxicity. We assessed ACV-induced nephrotoxicity and identified the differentially expressed proteins using mass spectrometry-based proteomic analysis. In total, 30 ICR mice were intraperitoneally administrated ACV (150 or 600 mg/kg per day) for 9 days. After administration of ACV, levels of serum creatinine and urea nitrogen increased significantly. In addition, mouse kidneys exhibited histopathological changes and reduced expression levels of vascular endothelial growth factor (VEGF) and its receptor VEGFR2. In the proteomic analysis, more than 1,000 proteins were separated by two-dimensional polyacrylamide gel electrophoresis, and a total of 20 proteins were up- or down-regulated in the ACV group compared with the saline group. Among these, six proteins (MHC class II antigen, glyoxalase 1, peroxiredoxin 1, αB-crystallin, fibroblast growth factor receptor 1-IIIb, and cytochrome c oxidase subunit Vb) were identified in association with ACV-induced nephrotoxicity. These findings were confirmed by Western blotting analysis. The differential expression levels of α-BC, Prx1, Glo I and CcO Vb suggest that oxidative damage and mitochondrial injury may be involved in ACV-induced nephrotoxicity. Furthermore, VEGF and FGF may play a role in tissue repair and the restoration process following ACV nephrotoxicity.


The critical influence of the intermediate category on interpretation errors in revised EUCAST and CLSI antimicrobial susceptibility testing guidelines.
Erroneous assignments of clinical isolates to the interpretative categories susceptible, intermediate and resistant can deprive a patient of successful antimicrobial therapy. The rate of major errors (ME) and very major errors (vME) is dependent on: (i) the precision/standard deviation (σ) of the antibiotic susceptibility testing (AST) method, (ii) the diameter distributions, (iii) clinical breakpoints, and (iv) the width of the intermediate zone. The European Committee on AST (EUCAST) has abandoned or decreased the intermediate zone for several drug/species combinations. This study focused on the effects of discontinuing the intermediate category on the rate of interpretation errors. In total, 10,341 non-duplicate clinical isolates were included in the study. For susceptibility testing the disc diffusion method was used. Error probabilities were calculated separately for diameter values flanking the interpretative category borders. Error probabilities were then applied to the actual numbers of clinical isolates investigated and expected rates of ME and vME were calculated. Applying EUCAST AST guidelines, significant rates of ME/vME were demonstrated for all drug/species combinations without an intermediate range. Virtually all ME/vME expected were eliminated in CLSI guidelines that retained an intermediate zone. If wild-type and resistant isolates are not clearly separated in susceptibility distributions, the retaining of an intermediate zone will decrease the number of ME and vME. An intermediate zone of 2-3 mm avoids almost all ME/vME for most species/drug combinations depending on diameter distributions. Laboratories should know their epidemiology settings to be able to detect problems of individual species/drug/clinical breakpoint combinations and take measures to improve precision of diameter measurements.


Male adolescents' reactions to TV beer advertisements: the effects of sports content and programming context.
This study examines white male adolescent responses to TV beer advertisements with and without sports content and to nonbeer ads when embedded in sports and entertainment programming. A total of 72 advertisements and 24 television program excerpts were randomly sampled from national television programming. White male adolescents (N = 157) recruited in a public school system each viewed six ads (one of each of three types of ad embedded in each of two types of programming) comprising the 2 x 2 x 3 factorial, within-subjects, mixed-model (random and fixed effects) experimental design along with an age-level blocking factor and random factors for commercial and program stimuli. Cognitive responses to each ad were content-analyzed. Individual difference variables including alcohol use behavior, sensation-seeking, masculinity and sports involvement were also measured. Subjects showed a consistent preference for beer ads with sports content. A significant three-way interaction between ad type, programming type and junior versus senior high-school age level also indicated that sports programming had an inconsistent effect on responses to beer ads but that nonbeer ads were responded to more positively during sports than during entertainment programming. Other analyses showed that subjects were more cognitively resistant to beer ads than to nonbeer ads. These results support public and official concerns that sports content in beer ads increase the ads appeal to underage youth. They do not support hypothesized concerns that sports programming might prime adolescents to be more receptive to beer ads. Implications for alcohol education efforts are discussed.


From chemical embryology to nucleosome patterning - an interview with Roumen G. Tsanev.
Roumen Tsanev was a prominent Bulgarian scientist whose pioneering ideas about the role for chromatin in cell differentiation and development led him to propose the first hypothesis for epigenetic information based on a histone code. To test experimentally his ideas, Dr.Tsanev explored nucleosome structure and heterogeneity and generated seminal data on nucleosome segregation in replicating chromatin. Roumen Tsanev made significant contributions to the understanding of chromatin changes that underlie zygotic gene activation. He identified sperm specific chromatin components tightly bound to DNA and demonstrated that the histone complement of the male pronucleus appears before the onset of DNA synthesis in the mouse zygote. In this interview, Roumen Tsanev talks about his passion for science and literature, reminisces about surmounting the harsh realities in post-war communist Bulgaria through creativity and determination, and explains what led him to propose that histones were carriers of epigenetic information. Dr. Tsanev discusses mathematical models of gene regulation and recalls computer simulations that reveal the non-linearity of genetic networks. He explains how this non-linearity could affect cell proliferation, differentiation, development and evolution.


An expanded mouse testis transcriptome and mass spectrometry defines novel proteins.
The testis transcriptome is exceptionally complex. Despite its complexity, previous testis transcriptome analyses relied on a reductive method for transcript identification, thus underestimating transcriptome complexity. We describe here a more complete testis transcriptome generated by combining Tuxedo, a reductive method, and spliced-RUM, a combinatorial transcript-building approach. Forty-two percent of the expanded testis transcriptome is composed of unannotated RNAs with novel isoforms of known genes and novel genes constituting 78 and 9.8% of the newly discovered transcripts, respectively. Across tissues, novel transcripts were predominantly expressed in the testis with the exception of novel isoforms which were also highly expressed in the adult ovary. Within the testis, novel isoform expression was distributed equally across all cell types while novel genes were predominantly expressed in meiotic and post-meiotic germ cells. The majority of novel isoforms retained their protein-coding potential while most novel genes had low protein-coding potential. However, a subset of novel genes had protein-coding potentials equivalent to known protein-coding genes. Shotgun mass spectrometry of round spermatid total protein identified unique peptides from four novel genes along with seven annotated non-coding RNAs. These analyses demonstrate the testis expresses a wide range of novel transcripts that give rise to novel proteins.