The Roles of GmERF135 in Improving Salt Tolerance and Decreasing ABA Sensitivity in Soybean.
Abscisic acid (ABA) mediates various abiotic stress responses, and ethylene responsive factors (ERFs) play vital role in resisting stresses, but the interaction of these molecular mechanisms remains elusive. In this study, we identified an ABA-induced soybean ERF gene GmERF135 that was highly up-regulated by ethylene (ET), drought, salt, and low temperature treatments. Subcellular localization assay showed that the GmERF135 protein was targeted to the nucleus. Promoter cis-acting elements analysis suggested that numerous potential stress responsive cis-elements were distributed in the promoter region of GmERF135, including ABA-, light-, ET-, gibberellin (GA)-, and methyl jasmonate (MeJA)-responsive elements. Overexpression of GmERF135 in Arabidopsis enhanced tolerance to drought and salt conditions. In addition, GmERF135 promoted the growth of transgenic hairy roots under salt and exogenous ABA conditions. These results suggest that soybean GmERF135 may participate in both ABA and ET signaling pathways to regulate the responses to multiple stresses.


How do we use in vitro models to understand epileptiform and ictal activity? A report of the TASK1-WG4 group of the ILAE/AES Joint Translational Task Force.
In vitro brain tissue preparations allow the convenient and affordable study of brain networks and have allowed us to garner molecular, cellular, and electrophysiologic insights into brain function with a detail not achievable in vivo. Preparations from both rodent and human postsurgical tissue have been utilized to generate in vitro electrical activity similar to electrographic activity seen in patients with epilepsy. A great deal of knowledge about how brain networks generate various forms of epileptiform activity has been gained, but due to the multiple in vitro models and manipulations used, there is a need for a standardization across studies. Here, we describe epileptiform patterns generated using in vitro brain preparations, focusing on issues and best practices pertaining to recording, reporting, and interpretation of the electrophysiologic patterns observed. We also discuss criteria for defining in vitro seizure-like patterns (i.e., ictal) and interictal discharges. Unifying terminologies and definitions are proposed. We suggest a set of best practices for reporting in vitro studies to favor both efficient across-lab comparisons and translation to in vivo models and human studies.


The variability, arrangement, and rearrangement of immunoglobulin genes.
The multiplicity of heavy-chain variable-region (VH) genes in mouse and human DNA has been estimated using a mouse heavy-(H) chain cDNA clone. We found about 10 hybridization components in mouse DNA and about 20 components in human DNA. Cross-hybridization studies of variable region (V) genes indicate that these components represent the numbers of genes within the VH subgroups of each of these species. The arrangement and rearrangement of the H-chain gamma subclasses have been studied in order to assess possible mechanisms of the H-chain switch. Evidence has been found for rearrangement events involving the gamma 2a and gamma 2b constant-region (CH) genes in DNA from cells making IgG2a and IgG2b respectively. In addition we found that cells making IgG2a lack detectable genes for gamma1 and gamma 2b. Both sets of observations are discussed in relation to H-chain diversity and the switch.


An exploratory study of the views of community pharmacy staff on the management of patients with undiagnosed skin problems.
The aim of this study was to explore pharmacist and medicine counter assistant (MCA) perceptions of community pharmacy management of patients presenting with symptomatic skin problems. The study involved semi-structured telephone interviews with 10 pharmacists and 15 MCAs from seven pharmacies. Interviews focused on perceptions of their role in managing symptomatic skin problems and views on why people sought pharmacy advice and any barriers to management. Advice from the local ethics committee was that approval was not required for the study. We analysed the interviews using the framework approach. Pharmacists identified two key themes that defined their role; triage and reassurance. In contrast, MCAs defined their role as information gatherers and independent advisors. Themes identified by both pharmacists and MCAs relating to the use of pharmacy as a source of advice were convenience, the perceived non-serious nature of conditions and inaccessibility of the GP. Additionally, MCAs believed familiarity with the pharmacist was important. Both pharmacists and MCAs identified their lack of dermatological knowledge as a barrier with pharmacists reporting insufficient time to deal effectively with patients. MCAs identified the potential for misdiagnosis by pharmacists as another potential barrier. Our findings suggest that the role of pharmacists and MCAs is complementary; MCAs screen and provide the necessary information to pharmacists who then decide upon an appropriate course of action for the patient. Nevertheless, a major barrier to pharmacy-supported self-care of symptomatic skin problems is a perceived lack of knowledge and training in dermatology.


Crayfish escape behavior: production of tailflips without giant fiber activity.
The giant interneurons of the crayfish nerve cord are well-known mediators of fast tail flexions, "tailflips," that propel animals through the water away from danger. More recent studies have revealed an additional nongiant generator of tailflips. In contrast to giant tailflips, which are stereotyped, nongiant tailflips have variable form. The operating principles and portions of the neural circuitry governing nongiant tailflips were here investigated. Whereas fast flexor motor neurons (FFs) receive excitatory postsynaptic potentials (EPSPs) with large unitary components just prior to giant tailflips, excitation of the FFs during nongiant tailflips is due to summation of many small EPSPs, and these build up for about 60 ms prior to the tailflip; we call the period of excitation prior to FF firing the preflexion phase and the period during which FFs fire, the flexion phase of the tailflip. Even FFs that will not fire during a given tailflip become depolarized during preflexion and flexion periods. Throughout the preflexion and flexion periods there is activity in dorsal nerve cord axons (DCAs) that lie below the giants. Many DCAs are interneurons that excite FFs at short latency. Some DCAs fire uniquely during the preflexion phase, while some fire only during the flexion phase. Which DCAs fire is highly variable, and in some cases firing of particular DCAs can be correlated with particular forms of tailflips. Two identified DCAs, 12 and 13, that fire during the flexion phase were studied. These interneurons originate and receive their synaptic input in the second and third abdominal ganglia, respectively, and project to the last ganglion exciting FFs caudal to their ganglion of origin en route. Their pattern of synaptic input prior to and during nongiant tailflips is indistinguishable from that of FFs. Their input to FFs is weak, but when they fire they tend to promote intersegmental synchrony of FFs in the segments they feed. It appears likely to us that nongiant tailflips are synthesized from a small library of component tailflip movements that can be combined to produce a variety of complete tailflips and that the component movements are produced by a limited group of premotor interneurons, of which 12 and 13 are members.(ABSTRACT TRUNCATED AT 400 WORDS)


Correlation between circulating endothelial cell counts and plasma thrombomodulin levels as markers for endothelial damage.
Increased numbers of circulating endothelial cells (CEC) in peripheral blood have been observed in diseases with vascular involvement, and are considered a promising surrogate marker for vascular damage. It was the objective of this study to evaluate the correlation between putative soluble markers of endothelial injury, activation, and endothelial proliferation, and absolute numbers of CEC. CEC were evaluated in 125 healthy donors and 40 patients with metastatic carcinoma by automated CD146 driven immunomagnetic isolation. Plasma concentrations of E-selectin, endoglin, and thrombomodulin were assessed by ELISA in plasma obtained from 40 healthy donors and 40 patients. CEC numbers in blood were positively correlated with plasma thrombomodulin levels, but not with levels of E-selectin and endoglin. Multivariate analysis demonstrated a significant increase in CEC numbers with age. The levels of plasma biomarkers were not influenced by age. Higher levels of thrombomodulin and E-selectin were observed in males when compared to females. In conclusion, CEC numbers correlate positively with plasma levels of thrombomodulin.


Enhancing the sensitivity of potential step voltammetry using chemometric resolution.
Enhancing the sensitivity of analytical methods by improving the signal quality is a universal goal in analytical chemistry. In analytical electrochemistry systems, the double layer charging current has been an obstacle to the accurate measurement of the faradaic current despite theoretical and experimental efforts. In this study, a method for sensitivity enhancement for potential step voltammetry is developed using chemometric resolution. Trilinear decomposition is used to extract the net faradaic current and double layer charging current directly from the data matrix of the decaying curves measured at different potentials. The feasibility of the method is proven using simulated data and further validated by two experimental datasets of diffusion and adsorption control reactions. Compared with the conventional approach that records the current data at a later pulse time, the voltammogram of the extracted net faradaic current is an ideal sigmoid curve with a horizontal baseline, even for the samples of very low concentration. More importantly, the analytical sensitivity can be greatly improved when the net faradaic current is used for quantitative determination.


Haploid yeast cells undergo a reversible phenotypic switch associated with chromosome II copy number.
SUP35 and SUP45 are essential genes encoding polypeptide chain release factors. However, mutants for these genes may be viable but display pleiotropic phenotypes which include, but are not limited to, nonsense suppressor phenotype due to translation termination defect. [PSI +] prion formation is another Sup35p-associated mechanism leading to nonsense suppression through decreased availability of functional Sup35p. [PSI +] differs from genuine sup35 mutations by the possibility of its elimination and subsequent re-induction. Some suppressor sup35 mutants had also been shown to undergo a reversible phenotypic switch in the opposite direction. This reversible switching had been attributed to a prion termed [ISP +]. However, even though many phenotypic and molecular level features of [ISP +] were revealed, the mechanism behind this phenomenon has not been clearly explained and might be more complex than suggested initially. Here we took a genomic approach to look into the molecular basis of the difference between the suppressor (Isp-) and non-suppressor (Isp+) phenotypes. We report that the reason for the difference between the Isp+ and the Isp- phenotypes is chromosome II copy number changes and support our finding with showing that these changes are indeed reversible by reproducing the phenotypic switch and tracking karyotypic changes. Finally, we suggest mechanisms that mediate elevation in nonsense suppression efficiency upon amplification of chromosome II and facilitate switching between these states. (i) In our experimental system, amplification of chromosome II confers nonsense suppressor phenotype and guanidine hydrochloride resistance at the cost of overall decreased viability in rich medium. (ii) SFP1 might represent a novel regulator of chromosome stability, as SFP1 overexpression elevates frequency of the additional chromosome loss in our system. (iii) Prolonged treatment with guanidine hydrochloride leads to selection of resistant isolates, some of which are disomic for chromosome II.


Early T Cell Recognition of B Cells following Epstein-Barr Virus Infection: Identifying Potential Targets for Prophylactic Vaccination.
Epstein-Barr virus, a B-lymphotropic herpesvirus, is the cause of infectious mononucleosis, has strong aetiologic links with several malignancies and has been implicated in certain autoimmune diseases. Efforts to develop a prophylactic vaccine to prevent or reduce EBV-associated disease have, to date, focused on the induction of neutralising antibody responses. However, such vaccines might be further improved by inducing T cell responses capable of recognising and killing recently-infected B cells. In that context, EBNA2, EBNA-LP and BHRF1 are the first viral antigens expressed during the initial stage of B cell growth transformation, yet have been poorly characterised as CD8+ T cell targets. Here we describe CD8+ T cell responses against each of these three "first wave" proteins, identifying target epitopes and HLA restricting alleles. While EBNA-LP and BHRF1 each contained one strong CD8 epitope, epitopes within EBNA2 induced immunodominant responses through several less common HLA class I alleles (e.g. B*3801 and B*5501), as well as subdominant responses through common class I alleles (e.g. B7 and C*0304). Importantly, such EBNA2-specific CD8+ T cells recognised B cells within the first day post-infection, prior to CD8+ T cells against well-characterised latent target antigens such as EBNA3B or LMP2, and effectively inhibited outgrowth of EBV-transformed B cell lines. We infer that "first wave" antigens of the growth-transforming infection, especially EBNA2, constitute potential CD8+ T cell immunogens for inclusion in prophylactic EBV vaccine design.


The relationship between social phobia and avoidant personality disorder: workshop report 3.
Social phobia and avoidant personality disorder were new diagnoses in the third version of the Diagnostic and Statistical Manual of Mental Disorders (DSM-III) and were considerably changed in the revised third version (DSM-III-R). A high rate of comorbidity between these disorders was observed, which can be explained by the use of the same diagnostic criteria for both conditions. While there has been considerable research activity on social phobia, little has been done on avoidant personality disorder. The relationship between these disorders, therefore, cannot be clearly stated at the moment. All of the empirical evidence so far supports a spectrum concept of social phobia with avoidant personality disorder rather than seeing the latter as a valid mental disorder.


Falling short of dietary guidelines - What do Australian pregnant women really know? A cross sectional study.
Maternal diets are not consistent with dietary guidance and this may affect the health of mothers and their infants. Nutrition knowledge and motivation may be important factors. To assess pregnant women's diets in relation to consistency with the Australian Guidelines for Healthy Eating (AGHE); factors influencing women's adherence to the recommendations; and women's attitudes towards pregnancy-specific nutrition information. A cross-sectional study using convenience sampling was undertaken at five hospitals in New South Wales (Australia) and through an online link (October 2012 to July 2013). N=388 pregnant women completed the survey. Categorical data were analysed using Chi square and logistic regression with significance set at P<0.05. Most participants were highly motivated to adopt a healthy diet, believed they were trying to do so and that knowing about nutrition in pregnancy was highly important. Reported dietary intakes were poor. No pregnant women met the recommended intakes for all five food groups. Poor knowledge of these recommendations was evident. Knowledge of selected recommendations (for Fruit, Vegetables, and Breads and Cereals) increased the likelihood of those foods' consumption 8 (95% confidence interval [CI], 2.3-27.7), 9.1 (95% CI, 2.6-31.3) and 6.8 (95% CI, 3.4-13.7) times respectively. Pregnant women had high levels of motivation and confidence in their ability to achieve a healthy diet and understand dietary recommendations, but actually demonstrated poor knowledge and poor adherence to guidelines. Mistaken or false beliefs may be a barrier to effective nutrition education strategies.


Comparison of eye-lens doses imparted during interventional and non-interventional neuroimaging techniques for assessment of intracranial aneurysms.
A neurointerventional examination of intracranial aneurysms often involves the eye lens in the primary beam of radiation. To assess and compare eye-lens doses imparted during interventional and non-interventional imaging techniques for the examination of intracranial aneurysms. We performed a phantom study on an anthropomorphic phantom (ATOM dosimetry phantom 702-D; CIRS, Norfolk, Virginia, USA) and assessed eye-lens doses with thermoluminescent dosimeters (TLDs) type 100 (LiF:Mg, Ti) during (1) interventional (depiction of all cerebral arteries with triple 3D-rotational angiography and twice 2-plane DSA anteroposterior and lateral projections) and (2) non-interventional (CT angiography (CTA)) diagnosis of intracranial aneurysms. Eye-lens doses were calculated following recommendations of the ICRP 103. Image quality was analysed in retrospective by two experienced radiologists on the basis of non-interventional and interventional pan-angiography examinations of patients with incidental aneurysms (n=50) on a five-point Likert scale. The following eye-lens doses were assessed: (1) interventional setting (triple 3D-rotational angiography and twice 2-plane DSA anteroposterior and lateral projections) 12 mGy; (2) non-interventional setting (CTA) 4.1 mGy. Image quality for depiction of intracranial aneurysms (>3 mm) was evaluated as good by both readers for both imaging techniques. Eye-lens doses are markedly higher during the interventional than during the non-interventional diagnosis of intracranial aneurysms. For the eye-lens dose, CTA offers considerable radiation dose savings in the diagnosis of intracranial aneurysms.


Risk appraisal and management of violent behavior.
To help clinicians enhance the safety of the public, hospital staff, and patients and improve patient management, this article briefly reviews recent empirical work on appraisal of the risk of violence and the management of violent individuals. Research on the prediction, management, and treatment of violent persons published in the last decade was reviewed. Risk appraisal research indicates that violence is predictable in some populations. The factors most highly and consistently related to risk are historical, including age, sex, past antisocial and violent conduct, psychopathy, aggressive childhood behavior, and substance abuse. Major mental disorder and psychiatric disturbance are poor predictors of violence. Actuarial methods are more accurate in predicting risk than unaided clinical judgment, which is a poor index. The Violence Risk Appraisal Guide has been shown to be a reliable and accurate actuarial instrument. Well-controlled studies have shown the effectiveness of behavior therapy and of behavioral staff training programs to reduce violence by persons in institutions, chronic psychiatric patients, and other populations. Otherwise, little is known about what psychotherapeutic or pharmacological treatments reduce violent recidivism by which clients under what circumstances. Recent work on the neurophysiology of aggression holds exciting promise but does not yet provide a scientific basis for prescriptive treatment. The most exciting and promising avenues for research on the management of violence lie in the joining of two scientific paths to understanding violence--biology and psychology.


First Report of Rice stripe virus of Proso Millet in Korea.
Rice stripe virus (RSV; genus Tenuivirus) is a serious threat to rice production in Korea (2). In 1965, a disease outbreak was observed on rice in South Korea, with plants showing yellow stripe symptoms (2). Reoccurrence of RSV in rice was observed again in 1980 in Gyeonggi and Chungcheong. In 2001, RSV was estimated to be infecting approximately 4,663 ha of rice in the provinces of Gyeonggi and Gangwha and approximately 5,000 ha of riceland in the provinces of Buan and, Seocheon (3). Proso millet (Panicum miliaceum L.) is grown as a cereal grain crop and used mainly for human food in South Korea (1). In June 2009, proso millet plants that displayed yellow stripe symptoms were collected at Sinjeon-Myeon, Gangjin-Gun, and Jeollanam-do provinces, where an outbreak of RSV in rice was reported. Diseased plants tested positive to RSV with an ELISA Kit (KisanBio, Seoul, Korea). Total RNA was extracted from leaf tissue with an RNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). RSV coat protein specific-primers were produced (5' TGTGGAACATAGTCCCACAGTAAGT 3'(upstream), 5' CTAAGCCGCAACCATTCCTCCAGT 3'(downstream). Reverse transcription-PCR confirmed the presence of a 494-bp product as predicted for the presence of RSV. The coat protein of RSV isolates collected from proso millet, rice, and foxtail millet in the same area was also sequenced. Results confirmed that phylogenetic relationships were of high homology: 98.9% between RSV isolates from rice and foxtail millet, 99.2% between isolates from rice (GenBank Accession No. JN245626) and proso millet (GenBank Accession No. JN245627); 99.6% between rice and foxtail millet (GenBank Accession No. JN245628); and 99.6% between foxtail millet and proso millet. In addition, sequence comparisons showed 96 to 99% identity with known RSV sequences available in GenBank (Accession No. X53563) (4). To our knowledge, this is the first report of RSV of proso millet in South Korea. The finding of this disease confirms further spread of the virus within the northern part of South Korea and the need for research to develop more effective management options to reduce the impact of RSV in proso millet. References: (1) Y. Y. Choi et al. Biosci. Biotechnol. Biochem. 69:31, 2005. (2) B. C. Lee et al. Res. Plant Dis. 10:30, 2004. (3) B. C. Lee et al. Res. Plant Dis. 14:210, 2008. (4) Y. Zhu. J. Gen. Virol. 72:763, 1991.


Development of a multi-layered psychosocial care system for children in areas of political violence.
Few psychosocial and mental health care systems have been reported for children affected by political violence in low- and middle income settings and there is a paucity of research-supported recommendations. This paper describes a field tested multi-layered psychosocial care system for children (focus age between 8-14 years), aiming to translate common principles and guidelines into a comprehensive support package. This community-based approach includes different overlapping levels of interventions to address varying needs for support. These levels provide assessment and management of problems that range from the social-pedagogic domain to the psychosocial, the psychological and the psychiatric domains. Specific intervention methodologies and their rationale are described within the context of a four-country program (Burundi, Sri Lanka, Indonesia and Sudan). The paper aims to contribute to bridge the divide in the literature between guidelines, consensus & research and clinical practice in the field of psychosocial and mental health care in low- and middle-income countries.


Dopaminergic enhancement of excitatory synaptic transmission in layer II entorhinal neurons is dependent on D₁-like receptor-mediated signaling.
The modulatory neurotransmitter dopamine induces concentration-dependent changes in synaptic transmission in the entorhinal cortex, in which high concentrations of dopamine suppress evoked excitatory postsynaptic potentials (EPSPs) and lower concentrations induce an acute synaptic facilitation. Whole-cell current-clamp recordings were used to investigate the dopaminergic facilitation of synaptic responses in layer II neurons of the rat lateral entorhinal cortex. A constant bath application of 1 μM dopamine resulted in a consistent facilitation of EPSPs evoked in layer II fan cells by layer I stimulation; the size of the facilitation was more variable in pyramidal neurons, and synaptic responses in a small group of multiform neurons were not modulated by dopamine. Isolated inhibitory synaptic responses were not affected by dopamine, and the facilitation of EPSPs was not associated with a change in paired-pulse facilitation ratio. Voltage-clamp recordings of α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) glutamate receptor-mediated excitatory postsynaptic currents (EPSCs) were facilitated by dopamine, but N-methyl-D-aspartate receptor-mediated currents were not. Bath application of the dopamine D₁-like receptor blocker SCH23390 (50 μM), but not the D₂-like receptor blocker sulpiride (50 μM), prevented the facilitation, indicating that it is dependent upon D₁-like receptor activation. Dopamine D₁ receptors lead to activation of protein kinase A (PKA), and including the PKA inhibitor H-89 or KT 5720 in the recording pipette solution prevented the facilitation of EPSCs. PKA-dependent phosphorylation of inhibitor 1 or the dopamine- and cAMP-regulated protein phosphatase (DARPP-32) can lead to a facilitation of AMPA receptor responses by inhibiting the activity of protein phosphatase 1 (PP1) that reduces dephosphorylation of AMPA receptors, and we found here that inhibition of PP1 occluded the facilitatory effect of dopamine. The dopamine-induced facilitation of AMPA receptor-mediated synaptic responses in layer II neurons of the lateral entorhinal cortex is therefore likely mediated via a D₁ receptor-dependent increase in PKA activity and a resulting inhibition in PP1-dependent dephosphorylation of AMPA receptors.


The balance between adhesion and contraction during cell division.
The ability to divide is a fundamental property of a living cell. The 3D orientation of cell division is essential for embryogenesis, maintenance of tissue organization and architecture, as well as controlling cell fate. Much attention has been placed on the mitotic spindle's role in placing itself along the cell's longest axis, where a shape sensing mechanism between a population of microtubules extending from mitotic centrosomes to the cell cortex occurs. However, contractile forces at the cell cortex also likely play a decisive role in determining the final placement of daughter cells following division. In this review, we discuss recent literature that describes the role of these contractile forces and how these forces could be balanced by mitotic adhesion complexes.


Zebrafish mutants identify an essential role for laminins in notochord formation.
Basement membranes are thought to be essential for organ formation, providing the scaffold on which individual cells organize to form complex tissues. Laminins are integral components of basement membranes. To understand the development of a simple vertebrate organ, we have used positional cloning to characterize grumpy and sleepy, two zebrafish loci known to control notochord formation, and find that they encode laminin beta1 and laminin gamma1, respectively. Removal of either chain results in the dramatic loss of laminin 1 staining throughout the embryo and prevents formation of the basement membrane surrounding the notochord. Notochord cells fail to differentiate and many die by apoptosis. By transplantation, we demonstrate that, for both grumpy and sleepy, notochord differentiation can be rescued by exogenous sources of the missing laminin chain, although notochordal sources are also sufficient for rescue. These results demonstrate a clear in vivo requirement for laminin beta1 and laminin gamma1 in the formation of a specific vertebrate organ and show that laminin or the laminin-dependent basement membrane is essential for the differentiation of chordamesoderm to notochord.


Mutations in the antifolate-resistance-associated genes dihydrofolate reductase and dihydropteroate synthase in Plasmodium vivax isolates from malaria-endemic countries.
Parasite dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) are known target enzymes of antifolate drugs used for the treatment and prophylaxis of persons with malaria. We sequenced the Plasmodium vivax dihydrofolate reductase (pvdhfr) and dihydropteroate synthase (pvdhps) genes to examine the prevalence and extent of point mutations in isolates from malaria-endemic countries. Double mutations (S58R and S117N) or quadruple mutations (F57L/I, S58R, T61M, and S117T) in the pvdhfr gene were found in isolates from Thailand (96.4%) and Myanmar (71.4%), but in only one isolate (1.0%) from Korea, where sulfadoxine-pyrimethamine has never been used. The pvdhfr point mutations correlated strongly with the pvdhps point mutations and ranged from single to triple mutations (S382A, A383G, and A553G), among isolates from Thailand, Myanmar, and Korea. These findings suggests that the prevalence of mutations in pvdhfr and pvdhps in P. vivax isolates from different malaria-endemic countries is associated with selection pressure imposed by sulfadoxine-pyrimethamine.


Intrathoracic paraganglioma arising from aorticosympathetic paraganglion.
A slowly growing intrathoracic paraganglion arising from an aorticosympathetic ganglion was removed from a 22-year-old woman. It was adherent to the parietal pleura and azygos vein, but not to the spinal cord. She has been well for the two years since that time. This patient represents the sixth tumor reported in this location. Most of the reported tumors are benign with occasional local invasion as seen in paragangliomas in other locations (carotid and aortic bodies). Complete excision of the tumor is the treatment of choice, but partial excision may provide long-term survival. A new classification and terminology suggested by Glenner and Grimley is emphasized. It groups logically the tumors arising from the paraganglion cells on the basis of anatomic distribution, innervation, and microscopic structure.


Fluorescence lifetime standards for time and frequency domain fluorescence spectroscopy.
A series of fluorophores with single-exponential fluorescence decays in liquid solution at 20 degrees C were measured independently by nine laboratories using single-photon timing and multifrequency phase and modulation fluorometry instruments with lasers as excitation source. The dyes that can serve as fluorescence lifetime standards for time-domain and frequency-domain measurements are all commercially available, are photostable under the conditions of the measurements, and are soluble in solvents of spectroscopic quality (methanol, cyclohexane, water). These lifetime standards are anthracene, 9-cyanoanthracene, 9,10-diphenylanthracene, N-methylcarbazole, coumarin 153, erythrosin B, N-acetyl-l-tryptophanamide, 1,4-bis(5-phenyloxazol-2-yl)benzene, 2,5-diphenyloxazole, rhodamine B, rubrene, N-(3-sulfopropyl)acridinium, and 1,4-diphenylbenzene. At 20 degrees C, the fluorescence lifetimes vary from 89 ps to 31.2 ns, depending on fluorescent dye and solvent, which is a useful range for modern pico- and nanosecond time-domain or mega- to gigahertz frequency-domain instrumentation. The decay times are independent of the excitation and emission wavelengths. Dependent on the structure of the dye and the solvent, the excitation wavelengths used range from 284 to 575 nm, the emission from 330 to 630 nm. These lifetime standards may be used to either calibrate or test the resolution of time- and frequency-domain instrumentation or as reference compounds to eliminate the color effect in photomultiplier tubes. Statistical analyses by means of two-sample charts indicate that there is no laboratory bias in the lifetime determinations. Moreover, statistical tests show that there is an excellent correlation between the lifetimes estimated by the time-domain and frequency-domain fluorometries. Comprehensive tables compiling the results for 20 (fluorescence lifetime standard/solvent) combinations are given.


Long-term survival and return on investment after nonneurologic injury: implications for the elderly trauma patient.
As the population of the United States ages and as the healthcare system undergoes significant change, cost effectiveness of care will become more important, particularly for older injured patients. The purpose of this study was to evaluate the cost per 2-year survivor stratified by age after moderate- to severe-nonneurologic injury. The trauma registry from a Level I trauma center was queried for adults (older than 18 years), discharged alive after blunt injury (Injury Severity Score >15), without significant neurologic injury, and with hospital charge data. Survival was determined using the Social Security Death Master File. Patients were stratified by age. Hospital costs were calculated by multiplying hospital charge by the cost to charge ratio. One thousand nine hundred fourteen patients made up the study population. Mean hospital cost per patient was $10,021. Mean cost per 2-year survivor was $10,328. Overall 2-year survival was 97%. (*p < 0.05 vs. youngest). When broken down by age group, there were no significant differences in hospital costs. However, 2-year survival was significantly less in those who were 55.1 years to 75 years old and those older than 75 years, when compared with those aged 18 years to 25 years. Thus, median cost per 2-year survivor was highest in those older than 75 years ($8,911). Although costs are similar by age at time of discharge, cost per 2-year survivor increases as age increases. However, cost per 2-year survivor does not exceed current cost-utility thresholds for any age group. Any future healthcare financing reforms should include aggressive funding for injury prevention efforts aimed at vulnerable populations instead of rationing care once an injury occurs.


Role of the Nox4/AMPK/mTOR signaling axe in adipose inflammation-induced kidney injury.
Diabetic kidney disease is one of the most serious complications of diabetes worldwide and is the leading cause of end-stage renal disease. While research has primarily focused on hyperglycemia as a key player in the pathophysiology of diabetic complications, recently, increasing evidence have underlined the role of adipose inflammation in modulating the development and/or progression of diabetic kidney disease. This review focuses on how adipose inflammation contribute to diabetic kidney disease. Furthermore, it discusses in detail the underlying mechanisms of adipose inflammation, including pro-inflammatory cytokines, oxidative stress, and AMPK/mTOR signaling pathway and critically describes their role in diabetic kidney disease. This in-depth understanding of adipose inflammation and its impact on diabetic kidney disease highlights the need for novel interventions in the treatment of diabetic complications.


Ours is human: on the pervasiveness of infra-humanization in intergroup relations.
Both at a conceptual and an empirical level, infra-humanization has been put on par with the relative greater attribution of uniquely human emotions to the in-group, assuming that a group's humanity is exclusively a matter of having uniquely human characteristics. In the present research we suggest that people also adopt another strategy to infra-humanize the out-group by considering those aspects that characterize and differentiate the in-group from the out-group as more uniquely human. In three studies, characteristics presented as typical of the in-group and the out-group were judged on a not uniquely human-uniquely human dimension. In addition to humanity, in Study 3 participants judged in-group and out-group characteristics also on an evaluative dimension. Consistent with the hypothesis, participants judged in-group characteristics as more human than those of the out-group, independent of their valence. The implications of these results for infra-humanization theory are discussed.


An update and systematic review on the treatment of primary dysmenorrhea.
Primary dysmenorrhea is a painful uterine contraction caused by endometrial laceration. Drug therapies and complementary medicine have been used to treat dysmenorrhea. The aim of this study was to investigate and offer an updated perspective on the treatments for dysmenorrhea. The present study was conducted in accordance with the PRISMA checklist for systematic reviews and meta-analyses. The required information was collected based on searches for the following keywords: treatment, primary dysmenorrhea, medicinal plants, chemical drugs, and herbs. Searches were performed on databases Pubmed, Web of Sciences, Scopus, Iran medex, and SID by March 2018 to find literature in the English and Persian languages on this subject without a time limit. This review included 17 papers, 10 of which on complementary medicine, three on drug therapies, and four on acupuncture and acupressure. The largest and smallest samples had 303 and 24 patients, respectively. Length of treatment ranged from one to six months and the measures most commonly used in the studies were the visual analogue scale and clinical efficacy. Reported complications included gastrointestinal events, nausea, vomiting, diarrhea, abdominal pain, and liver and kidney disorders. Medicinal plants, drugs, and acupressure seem to suppress pain by reducing the level of prostaglandins, mediating nitric oxide, increasing beta-endorphin levels, blocking the calcium channel, and enhancing circulatory flow through the uterine pathway. Further trials are required to confirm the benefits of the procedures described and ensure the absence of complications.


Self-care Strategies to Combat Burnout Among Pediatric Critical Care Nurses and Physicians.
Professional burnout is a widespread phenomenon in health care. The health of patients and organizations begins with the well-being of health care professionals. Identifying and understanding self-care strategies that professionals perceive to be helpful is crucial to combat burnout. To determine perceptions of self-care strategies to combat professional burnout among nurses and physicians in pediatric critical care settings. This was a qualitative descriptive study with a phenomenological overtone. The study was conducted in a 20-bed pediatric intensive care unit and an 8-bed intermediate care unit of a children's hospital in the United States. Information flyers and emails were used to introduce the study. A combination of convenience and purposive sampling methods was used to recruit participants who were full-time nurses and physicians in the 2 units. Information saturation was used to regulate sample sizes, resulting in 20 participants. Data were collected through a onetime face-to-face interview with each participant. A qualitative descriptive approach was used to analyze the data. The first author was the primary coder and discussed the codes with the coauthors throughout the coding process. Six major self-care strategies were identified: finding meaning in work, connecting with an energy source, nurturing interpersonal connections, developing an attitude of positivity, performing emotional hygiene, and recognizing one's uniqueness and contributions at work. Developing effective self-care strategies helps promote health care professionals' physical and psychological well-being and reduce burnout. It is vital for health care professionals to care for themselves so that they can best care for others.


Left Ventricular Diastolic Dysfunction in a Rat Model of Diabetic Cardiomyopathy using ECG-gated 18F-FDG PET.
In diabetic cardiomyopathy, left ventricular (LV) diastolic dysfunction is one of the earliest signs of cardiac involvement prior to the definitive development of heart failure (HF). We aimed to explore the LV diastolic function using electrocardiography (ECG)-gated 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET) imaging beyond the assessment of cardiac glucose utilization in a diabetic rat model. ECG-gated 18F-FDG PET imaging was performed in a rat model of type 2 diabetes (ZDF fa/fa) and ZL control rats at age of 13 weeks (n = 6, respectively). Under hyperinsulinemic-euglycemic clamp to enhance cardiac activity, 18F-FDG was administered and subsequently, list-mode imaging using a dedicated small animal PET system with ECG signal recording was performed. List-mode data were sorted and reconstructed into tomographic images of 16 frames per cardiac cycle. Left ventricular functional parameters (systolic: LV ejection fraction (EF), heart rate (HR) vs. diastolic: peak filling rate (PFR)) were obtained using an automatic ventricular edge detection software. No significant difference in systolic function could be obtained (ZL controls vs. ZDF rats: LVEF, 62.5 ± 4.2 vs. 59.4 ± 4.5%; HR: 331 ± 35 vs. 309 ± 24 bpm; n.s., respectively). On the contrary, ECG-gated PET imaging showed a mild but significant decrease of PFR in the diabetic rats (ZL controls vs. ZDF rats: 12.1 ± 0.8 vs. 10.2 ± 1 Enddiastolic Volume/sec, P < 0.01). Investigating a diabetic rat model, ECG-gated 18F-FDG PET imaging detected LV diastolic dysfunction while systolic function was still preserved. This might open avenues for an early detection of HF onset in high-risk type 2 diabetes before cardiac symptoms become apparent.


Cervical decompression and reconstruction without intraoperative neurophysiological monitoring.
The primary goal of this study was to review the immediate postoperative neurological function in patients surgically treated for symptomatic cervical spine disease without intraoperative neurophysiological monitoring. The secondary goal was to assess the economic impact of intraoperative monitoring (IOM) in this patient population. This study is a retrospective review of 720 consecutively treated patients who underwent cervical spine procedures. The patients were identified and the data were collected by individuals who were not involved in their care. A total of 1534 cervical spine levels were treated in 720 patients using anterior, posterior, and combined (360°) approaches. Myelopathy was present preoperatively in 308 patients. There were 185 patients with increased signal intensity within the spinal cord on preoperative T2-weighted MR images, of whom 43 patients had no clinical evidence of myelopathy. Three patients (0.4%) exhibited a new neurological deficit postoperatively. Of these patients, 1 had a preoperative diagnosis of radiculopathy, while the other 2 were treated for myelopathy. The new postoperative deficits completely resolved in all 3 patients and did not require additional treatment. The Current Procedural Terminology (CPT) codes for IOM during cervical decompression include 95925 and 95926 for somatosensory evoked potential monitoring of the upper and lower extremities, respectively, as well as 95928 and 95929 for motor evoked potential monitoring of the upper and lower extremities. In addition to the charge for the baseline [monitoring] study, patients are charged hourly for ongoing electrophysiology testing and monitoring using the CPT code 95920. Based on these codes and assuming an average of 4 hours of monitoring time per surgical case, the savings realized in this group of patients was estimated to be $1,024,754. With the continuing increase in health care costs, it is our responsibility as providers to minimize expenses when possible. This should be accomplished without compromising the quality of care to patients. This study demonstrates that decompression and reconstruction for symptomatic cervical spine disease without IOM may reduce the cost of treatment without adversely impacting patient safety.


The effect of binaural beat stimulation on sustained attention.
Binaural beats have been used as a way of modifying cognition via auditory stimulation. A recent meta-analysis suggests that binaural beat stimulation can have a positive effect on attention (Garcia-Argibay et al., Psychologische Forschung 83:1124-1136, 2019a, Psychological Research Psychologische Forschung 83:357-372, 2019b), with the sample-weighted average effect size being about .58. This is an intriguing and potentially useful finding, both theoretically and practically. In this study, we focus on sustained attention. We delivered beta-frequency (16 Hz) binaural beat stimulation during a sustained attention task. In "Experiment 1", reaction times were faster under beat stimulation than control stimulation in a between-subjects design. However, the effect was modest in magnitude, and model comparisons using Bayes Factors were indiscriminate between including and excluding the effect from the model. We followed this initial experiment with two concurrently administered follow-up experiments. In "Experiment 2", we added thought probes to measure any changes in task engagement associated with binaural beat stimulation. "Experiment 2" revealed a different effect from "Experiment 1": participants in the binaural beat condition exhibited a shallower vigilance decrement. However, the beat stimulation did not affect the thought probes responses. Combining data across the two experiments indicated rather strong evidence against the hypothesis that beta-frequency binaural beats can augment sustained attention, either via a general speeding of responding or a mitigation of the vigilance decrement. Finally, in "Experiment 3", we investigated whether pupillary measures of arousal and/or task engagement would be affected by binaural beat stimulation. There was no evidence for such effects. Overall, we did not observe any consistent evidence that binaural beat stimulation can augment sustained attention or its subjective and physiological correlates.


Disclosure of HIV serostatus and condomless sex among men living with HIV/AIDS in Florida.
Despite campaigns to increase safer sex practices, there are people living with HIV/AIDS (PLWH) who do not disclose their HIV status to sexual partners and engage in condomless sex. The purpose of this research was to: 1) describe factors associated with disclosure of HIV status to sexual partners; and 2) determine if disclosure and/or receipt of prevention counseling are independently associated with condomless sex. We used the Florida Medical Monitoring Project to analyze data from 376 HIV positive men with more than one sexual partner. Results indicated that 55% consistently disclosed their HIV status to sexual partners, 30% inconsistently disclosed, 15% did not disclose, and 48% reported any condomless sex. The odds of having condomless sex was 3.3 (CI = 1.5, 7.3) times greater in men who disclosed to all partners. Results suggest that men who disclose are also those who are more likely to have condomless sex. More research is needed to better understand the complex nature of disclosure and sexual risk behaviors and how disclosure impacts sexual risk.


Capsicum annuum tobacco mosaic virus-induced clone 1 expression perturbation alters the plant's response to ethylene and interferes with the redox homeostasis.
Capsicum annuum tobacco mosaic virus (TMV)-induced clone 1 (CaTin1) gene was expressed early during incompatible interaction of hot pepper (Caspsicum annuum) plants with TMV and Xanthomonas campestris. RNA-blot analysis showed that CaTin1 gene was expressed only in roots in untreated plants and induced mainly in leaf in response to ethylene, NaCl, and methyl viologen but not by salicylic acid and methyl jasmonate. The ethylene dependence of CaTin1 induction upon TMV inoculation was demonstrated by the decrease of CaTin1 expression in response to several inhibitors of ethylene biosynthesis or its action. Transgenic tobacco (Nicotiana tabacum) plants expressing CaTin1 gene in sense- or antisense-orientation showed interesting characteristics such as the accelerated growth and the enhanced resistance to biotic as well as abiotic stresses. Such characteristics appear to be caused by the elevated level of ethylene and H2O2. Moreover, in transgenic plants expressing antisense CaTin1 gene, the expression of some pathogenesis-related genes was enhanced constitutively, which may be mainly due to the increased ethylene level. The promoter of CaTin1 has four GCC-boxes, two AT-rich regions, and an elicitor-inducible W-box. The induction of the promoter activity by ethylene depends on GCC-boxes and by TMV on W-box. Taken together, we propose that the CaTin1 up-regulation or down-regulation interferes with the redox balance of plants leading to the altered response to ethylene and biotic as well as abiotic stresses.


Solution-phase library screening for the identification of rare clones: isolation of an alpha 1D-adrenergic receptor cDNA.
alpha 1-Adrenergic receptor (alpha 1-AR) subtypes (alpha 1A and alpha 1B) play a critical role in vascular smooth muscle contraction and circulatory homeostasis. Transcripts for these guanine nucleotide-binding protein-coupled receptors are extremely low in abundance, however, and isolation of their cDNAs is difficult. We have developed a novel technique for identifying rare clones in a cDNA library, which has been used successfully to isolate a cDNA clone encoding an alpha 1D-AR. A 564-bp polymerase chain reaction product encoding a region between the third and sixth transmembrane domains of the alpha 1D-AR was first generated using rat brain mRNA as template and highly degenerate primers. The primers corresponded to those domains but contained mismatches to the alpha 1B-AR sequences. A 3-kb transcript was identified with this polymerase chain reaction probe, by Northern analysis of rat hippocampus. However, traditional plaque hybridization failed to identify a cDNA in a rat hippocampus lambda gt10 library. By solution-phase screening of virtually the entire library, a cDNA containing a 3-kb insert was identified, amplified, and purified. This insert encodes a 560-amino acid protein corresponding to the topology of guanine nucleotide-binding protein-coupled receptors. This receptor has approximately 71% amino acid identity, in the transmembrane regions, to the hamster and rat alpha 1B-ARs. Characterization of the receptor expressed in COS-7 cells, by ligand binding and photoaffinity labeling, revealed some of the characteristics of an alpha 1A-AR. However, unlike alpha 1A-ARs characterized previously in membrane preparations or in solubilized partially purified preparations, the expressed receptor could be extensively inactivated by chlorethylclonidine. In addition, it displays ligand-binding properties that are not consistent with an alpha 1A-AR. This indicates that the cDNA clone that we have isolated encodes a novel alpha 1-AR subtype, which we classify as the alpha 1D-AR.


Frailty status can be accurately assessed using inertial sensors and the TUG test.
frailty is an important geriatric syndrome linked to increased mortality, morbidity and falls risk. a total of 399 community-dwelling older adults were assessed using Fried's frailty phenotype and the timed up and go (TUG) test. Tests were quantified using shank-mounted inertial sensors. We report a regression-based method for assessment of frailty using inertial sensor data obtained during TUG. For comparison, frailty was also assessed using the same method based on grip strength and manual TUG time. using inertial sensor data, participants were classified as frail or non-frail with mean accuracy of 75.20% (stratified by gender). Using TUG time alone, frailty status was classified correctly with mean classification accuracy of 71.82%. Similarly, using grip strength alone, the frailty status was classified correctly with mean classification accuracy of 77.65%. Stratifying sensor data by gender yielded significantly (p<0.05) increased accuracy in classifying frailty when compared with equivalent manual TUG time-based models. results suggest that a simple protocol involving assessment using a well-known mobility test (Timed Up and Go (TUG)) and inertial sensors can be a fast and effective means of automatic, non-expert assessment of frailty.


Targeting the diuretic hormone receptor to control the cotton leafworm, Spodoptera littoralis.
The cotton leafworm, Spodoptera littoralis Boisduval (Lepidoptera: Noctuidae), is one of the most devastating pests of crops worldwide. Several types of treatments have been used against this pest, but many of them failed because of the rapid development of genetic resistance in the different insect populations. G protein coupled receptors have vital functions in most organisms, including insects; thus, they are appealing targets for species-specific pest control strategies. Among the insect G protein coupled receptors, the diuretic hormone receptors have several key roles in development and metabolism, but their importance in vivo and their potential role as targets of novel pest control strategies are largely unexplored. With the goal of using DHR genes as targets to control S. littoralis, we cloned a corticotropin-releasing factor-like binding receptor in this species and expressed the corresponding dsRNA in tobacco plants to knock down the receptor activity in vivo through RNA interference. We also expressed the receptor in mammalian cells to study its signaling pathways. The results indicate that this diuretic hormone receptor gene has vital roles in S. littoralis and represents an excellent molecular target to protect agriculturally-important plants from this pest.


Visual hallucinations and sight loss in children and young adults: a retrospective case series of Charles Bonnet syndrome.
Charles Bonnet syndrome (CBS) is a complication of sight loss affecting all ages; yet, few childhood cases have been reported. Our aim is to raise awareness of this under-reported association occurring in children and young adults in order to prevent psychological harm in this age group. A retrospective case series reviewing medical notes of patients <25 years of age with sight loss and reported CBS at a single centre hospital eye service in London, United Kingdom. Search of electronic patient records identified 13 patients experiencing hallucinatory events over a 9-year period. Outcomes were patient demographics including ocular diagnosis, visual acuity at time of onset, characteristics of hallucinations, clinical management strategies and patient-reported affliction. Eight patients were diagnosed with progressive inherited retinal diseases, primarily Stargardt disease (N=5). Clinical history indicated patients had significantly reduced best-corrected visual acuity (BCVA) in at least one eye at onset; median (IQR) worse eye BCVA was 1.0 (0.86-1.6) logarithm of minimum angle of resolution(LogMAR). CBS significantly affected patients' personal lives including education, diet and sleep. Clinical management was varied, mostly relating to reassurance at the point of contact. We describe the clinical features of young patients with CBS, with management strategies and aspects of negative outcomes. High potential caseload and risk of psychological harm merit further research. Increased awareness among healthcare professionals and patient education to forewarn susceptible individuals may reduce the overall impact and improve coping with symptoms.


Effect of physical exercise on physical performance and fall incidents of individuals living with dementia in nursing homes: a randomized controlled trial.
Objective: To determine the effects of an exercise intervention on physical performance and reported fall incidents among individuals living with dementia in nursing homes. Methods: The study was a randomized controlled trial to determine the effect of the physical activity on physical performance and reported fall incidents. A sample of 60 participants from two nursing homes in Australia were randomly allocated to either: (1) Intervention Group 1: physical exercise intervention for 45 min, once a week; (2) Intervention Group 2: physical exercise intervention for 15 min, three times a week; or (3) usual care Control Group. Physical performance was assessed before and after the intervention (12 weeks) using: Six Meter Walk test, Five-Times-Sit-to-Stand test, Timed Up and Go (TUG) test, (Modified) Functional Reach test, timed static pedaling (TSP). The number of reported falls was determined by review of incident reports completed by nursing home staff. Results: The physical performance outcome measures demonstrated positive trends over time in favor of the intervention groups, though the observed changes were only statistically significant for TSP and TUG Test. The number of reported falls demonstrated a significant difference between groups during the intervention period. Conclusions: The results demonstrated that the physical exercise intervention could be beneficial for individuals living with dementia, and as little as 45 min per week could be effective for this population group. However, cautious interpretation was drawn as the pool of participants was not sufficiently large enough to generate a meaningful effect size.


Study protocol for the Integra Initiative to assess the benefits and costs of integrating sexual and reproductive health and HIV services in Kenya and Swaziland.
In sub-Saharan Africa (SSA) there are strong arguments for the provision of integrated sexual and reproductive health (SRH) and HIV services. Most HIV transmissions are sexually transmitted or associated with pregnancy, childbirth, and breastfeeding. Many of the behaviours that prevent HIV transmission also prevent sexually transmitted infections and unintended pregnancies. There is potential for integration to increase the coverage of HIV services, as individuals who use SRH services can benefit from HIV services and vice-versa, as well as increase cost-savings. However, there is a dearth of empirical evidence on effective models for integrating HIV/SRH services. The need for robust evidence led a consortium of three organizations - International Planned Parenthood Federation, Population Council and the London School of Hygiene & Tropical Medicine - to design/implement the Integra Initiative. Integra seeks to generate rigorous evidence on the feasibility, effectiveness, cost and impact of different models for delivering integrated HIV/SRH services in high and medium HIV prevalence settings in SSA. A quasi-experimental study will be conducted in government clinics in Kenya and Swaziland - assigned into intervention/comparison groups. Two models of service delivery are investigated: integrating HIV care/treatment into 1) family planning and 2) postnatal care. A full economic-costing will be used to assess the costs of different components of service provision, and the determinants of variations in unit costs across facilities/service models. Health facility assessments will be conducted at four time-periods to track changes in quality of care and utilization over time. A two-year cohort study of family planning/postnatal clients will assess the effect of integration on individual outcomes, including use of SRH services, HIV status (known/unknown) and pregnancy (planned/unintended). Household surveys within some of the study facilities' catchment areas will be conducted to profile users/non-users of integrated services and demand/receipt of integrated services, before-and-after the intervention. Qualitative research will be conducted to complement the quantitative component at different time points. Integra takes an embedded 'programme science' approach to maximize the uptake of findings into policy/practice. Integra addresses existing evidence gaps in the integration evaluation literature, building on the limited evidence from SSA and the expertise of its research partners. Current Controlled Trials NCT01694862.


Ketamine analgesia for inflammatory pain in neonatal rats: a factorial randomized trial examining long-term effects.
Neonatal rats exposed to repetitive inflammatory pain have altered behaviors in young adulthood, partly ameliorated by Ketamine analgesia. We examined the relationships between protein expression, neuronal survival and plasticity in the neonatal rat brain, and correlated these changes with adult cognitive behavior. Using Western immunoblot techniques, homogenates of cortical tissue were analyzed from neonatal rats 18-20 hours following repeated exposure to 4% formalin injections (F, N = 9), Ketamine (K, 2.5 mg/kg x 2, N = 9), Ketamine prior to formalin (KF, N = 9), or undisturbed controls (C, N = 9). Brain tissues from another cohort of rat pups (F = 11, K = 12, KF = 10, C = 15) were used for cellular staining with Fos immunohistochemistry or FluoroJade-B (FJB), followed by cell counting in eleven cortical and three hippocampal areas. Long-term cognitive testing using a delayed non-match to sample (DNMS) paradigm in the 8-arm radial maze was performed in adult rats receiving the same treatments (F = 20, K = 24, KF = 21, C = 27) in the neonatal period. Greater cell death occurred in F vs. C, K, KF in parietal and retrosplenial areas, vs. K, KF in piriform, temporal, and occipital areas, vs. C, K in frontal and hindlimb areas. In retrosplenial cortex, less Fos expression occurred in F vs. C, KF. Cell death correlated inversely with Fos expression in piriform, retrosplenial, and occipital areas, but only in F. Cortical expression of glial fibrillary acidic protein (GFAP) was elevated in F, K and KF vs. C. No significant differences occurred in Caspase-3, Bax, and Bcl-2 expression between groups, but cellular changes in cortical areas were significantly correlated with protein expression patterns. Cluster analysis of the frequencies and durations of behaviors grouped them as exploratory, learning, preparatory, consumptive, and foraging behaviors. Neonatal inflammatory pain exposure reduced exploratory behaviors in adult males, learning and preparatory behaviors in females, whereas Ketamine ameliorated these long-term effects. Neuroprotective effects of Ketamine attenuate the impaired cognitive behaviors resulting from pain-induced cell death in the cortical and hippocampal fields of neonatal rats. This cell death was not dependent on the apoptosis associated proteins, but was correlated with glial activation.


Sulfotransferase 1A1 polymorphism, endogenous estrogen exposure, well-done meat intake, and breast cancer risk.
Sulfotransferase (SULT) 1A1 is involved in the inactivation of estrogens and bioactivation of heterocyclic amines and polycyclic aromatic hydrocarbons. A G--->A transition at codon 213 (CGC/Arg to CAC/His) of the SULT1A1 gene was reported recently, and individuals homozygous for the His allele have a substantially lower activity of this enzyme than those with other genotypes. We hypothesized that the His allele may be a risk factor for breast cancer, particularly among women who had risk factors related to higher endogenous estrogen exposure. This hypothesis was investigated in a case-control study conducted in a cohort of postmenopausal Iowa women who completed a mailed questionnaire in 1986 on lifestyle factors including information on major breast cancer risk factors. DNA samples and information related to well-done meat intake were obtained from breast cancer cases diagnosed between 1992 and 1994 and a random sample of cancer-free cohort members. Multivariate analysis was performed on data from 156 cases and 332 controls who donated a blood sample. The frequency of the His allele was 41.6% in cases and 34.1% in controls (P = 0.03), and the risk of breast cancer was increased with the number of His alleles (P for trend = 0.02). Compared with women with the Arg/Arg genotype, an 80% elevated risk was observed among women homozygous for the His allele (95% confidence interval, 1.0-3.2; P = 0.04). This positive association was more pronounced among women who drank alcohol and had a high body mass index, early age at menarche, and late age at menopause, factors related to high endogenous estrogen exposure, than among those who did not have these risk factors. The risk of breast cancer was elevated with increasing doneness level of red meat intake among women with the Arg/Arg genotype (P for trend, 0.01) or the Arg/His genotype (P for trend, 0.10), whereas this association was not evident for women with the His/His genotype. The results from this study suggest that homozygosity for the SULT1A1 His213 allele may be a risk factor for breast cancer, and its effect may be modified by the exposure level of endogenous estrogens and heterocyclic amines.


Chemometrics applications in biotechnology processes: predicting column integrity and impurity clearance during reuse of chromatography resin.
Separation media, in particular chromatography media, is typically one of the major contributors to the cost of goods for production of a biotechnology therapeutic. To be cost-effective, it is industry practice that media be reused over several cycles before being discarded. The traditional approach for estimating the number of cycles a particular media can be reused for involves performing laboratory scale experiments that monitor column performance and carryover. This dataset is then used to predict the number of cycles the media can be used at manufacturing scale (concurrent validation). Although, well accepted and widely practiced, there are challenges associated with extrapolating the laboratory scale data to manufacturing scale due to differences that may exist across scales. Factors that may be different include: level of impurities in the feed material, lot to lot variability in feedstock impurities, design of the column housing unit with respect to cleanability, and homogeneity of the column packing. In view of these challenges, there is a need for approaches that may be able to predict column underperformance at the manufacturing scale over the product lifecycle. In case such an underperformance is predicted, the operators can unpack and repack the chromatography column beforehand and thus avoid batch loss. Chemometrics offers one such solution. In this article, we present an application of chemometrics toward the analysis of a set of chromatography profiles with the intention of predicting the various events of column underperformance including the backpressure buildup and inefficient deoxyribonucleic acid clearance.


Biochemical factors modulating cellular neurotoxicity of methylmercury.
Methylmercury (MeHg), an environmental toxicant primarily found in fish and seafood, poses a dilemma to both consumers and regulatory authorities, given the nutritional benefits of fish consumption versus the possible adverse neurological damage. Several studies have shown that MeHg toxicity is influenced by a number of biochemical factors, such as glutathione (GSH), fatty acids, vitamins, and essential elements, but the cellular mechanisms underlying these complex interactions have not yet been fully elucidated. The objective of this paper is to outline the cellular response to dietary nutrients, as well as to describe the neurotoxic exposures to MeHg. In order to determine the cellular mechanism(s) of toxicity, the effect of pretreatment with biochemical factors (e.g., N-acetyl cysteine, (NAC); diethyl maleate, (DEM); docosahexaenoic acid, (DHA); selenomethionine, SeM; Trolox) and MeHg treatment on intercellular antioxidant status, MeHg content, and other endpoints was evaluated. This paper emphasizes that the protection against oxidative stress offered by these biochemical factors is among one of the major mechanisms responsible for conferring neuroprotection. It is therefore critical to ascertain the cellular mechanisms associated with various dietary nutrients as well as to determine the potential effects of neurotoxic exposures for accurately assessing the risks and benefits associated with fish consumption.


Stem Cells in Orthopedics: A Comprehensive Guide for the General Orthopedist.
The use of biologic adjuvants in the treatment of operative and nonoperative orthopedic injuries continues to expand in concert with our understanding of the acute and chronic healing process of musculoskeletal injuries. Stem cell treatments in orthopedics are among the most commonly explored options, and have found varying levels of success in promoting osseous and soft tissue healing. Basic science and translational studies have demonstrated the potential for broad application of stem cells in the treatment of a growing number of musculoskeletal injuries. Emerging clinical studies have also provided promising results, although the vast majority of studies have featured small sample sizes and limited duration of follow-up. In addition, a number of important questions remain regarding the clinical safety, treatment delivery, and overall efficacy of stem cell augmentation of injured tissue in orthopedics. The objective of the current review is to present a broad overview of the current state of stem cell treatments in orthopedic surgery, with an emphasis on soft tissue healing. This review of stem cell treatment covers the basic science behind biologic augmentation, advantages of the various stem cell sources, preclinical results, and current and future clinical applications.


Peptide inhibitors of C3 activation as a novel strategy of complement inhibition for the treatment of paroxysmal nocturnal hemoglobinuria.
Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by complement-mediated intravascular hemolysis due to the lack of CD55 and CD59 on affected erythrocytes. The anti-C5 antibody eculizumab has proven clinically effective, but uncontrolled C3 activation due to CD55 absence may result in opsonization of erythrocytes, possibly leading to clinically meaningful extravascular hemolysis. We investigated the effect of the peptidic C3 inhibitor, compstatin Cp40, and its long-acting form (polyethylene glycol [PEG]-Cp40) on hemolysis and opsonization of PNH erythrocytes in an established in vitro system. Both compounds demonstrated dose-dependent inhibition of hemolysis with IC50 ∼4 µM and full inhibition at 6 µM. Protective levels of either Cp40 or PEG-Cp40 also efficiently prevented deposition of C3 fragments on PNH erythrocytes. We further explored the potential of both inhibitors for systemic administration and performed pharmacokinetic evaluation in nonhuman primates. A single intravenous injection of PEG-Cp40 resulted in a prolonged elimination half-life of >5 days but may potentially affect the plasma levels of C3. Despite faster elimination kinetics, saturating inhibitor concentration could be reached with unmodified Cp40 through repetitive subcutaneous administration. In conclusion, peptide inhibitors of C3 activation effectively prevent hemolysis and C3 opsonization of PNH erythrocytes, and are excellent, and potentially cost-effective, candidates for further clinical investigation.


Reversal of doxorubicin resistance by hydrophobic, but not hydrophilic, forskolins.
The plant diterpene forskolin reverses acquired resistance to doxorubicin in variants of the murine sarcoma S180 cell line. Because forskolin is known to elevate intracellular cAMP levels, investigations were performed to determine whether this reversal of resistance resulted from effects on signal transduction. Two analogues of forskolin, dideoxyforskolin, which does not elevate cAMP, and a water-soluble analogue, were also investigated. Although all three diterpenes elevated levels of either cAMP or protein kinase C, these effects were not consistently associated with reversal of doxorubicin resistance. Likewise, all three diterpenes were capable of displacing [3H]azidopine from P-glycoprotein, but reversal of doxorubicin resistance was observed only with forskolin and dideoxyforskolin, suggesting that binding to P-glycoprotein may be a necessary, but not sufficient, condition for reversing doxorubicin resistance. The hydrophobicity of the compounds appeared to be the single factor most consistently related to reversal of doxorubicin resistance in this cell system, with the hydrophilic compound water-soluble forskolin failing to produce this result, even at concentrations 10-fold higher than effective concentrations of the hydrophobic diterpenes.


Keratinocyte expression of inflammatory mediators plays a crucial role in substance P-induced acute and chronic pain.
Tibia fracture in rats followed by cast immobilization leads to nociceptive, trophic, vascular and bone-related changes similar to those seen in Complex Regional Pain Syndrome (CRPS). Substance P (SP) mediated neurogenic inflammation may be responsible for some of the signs of CRPS in humans. We therefore hypothesized that SP acting through the SP receptor (NK1) leads to the CRPS-like changes found in the rat model. In the present study, we intradermally injected rats with SP and monitored hindpaw mechanical allodynia, temperature, and thickness as well as tissue levels of tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), interleukin 6 (IL-6), and nerve growth factor-β (NGF) for 72 h. Anti-NGF antibody was utilized to block the effects of SP-induced NGF up-regulation. Fracture rats treated with the selective NK1 receptor antagonist LY303870 prior to cast removal were assessed for BrdU, a DNA synthesis marker, incorporation in skin cells to examine cellular proliferation. Bone microarchitecture was measured using micro computed tomography (μCT). We observed that: (1) SP intraplantar injection induced mechanical allodynia, warmth and edema as well as the expression of nociceptive mediators in the hindpaw skin of normal rats, (2) LY303870 administered intraperitoneally after fracture attenuated allodynia, hindpaw unweighting, warmth, and edema, as well as cytokine and NGF expression, (3) LY303870 blocked fracture-induced epidermal thickening and BrdU incorporation after fracture, (4) anti-NGF antibody blocked SP-induced allodynia but not warmth or edema, and (5) LY303870 had no effect on bone microarchitecture. Collectively our data indicate that SP acting through NK1 receptors supports the nociceptive and vascular components of CRPS, but not the bone-related changes.


Intraoperative complications of laparoscopic adrenalectomy.
A laparoscopic or retroperitoneoscopic access to the adrenal gland is the standard of care for adrenalectomy in most cases. Although in laparoscopic adrenalectomy the approach is minimally invasive, the procedure is challenging. This is reflected in the scope of possible complications. The surgeon must consider complications related to the anatomical topography of the adrenal gland, which typically encompasses the complications known from open surgery and complications related to the minimal invasive access. In this topic paper we will address the most frequently encountered complications of adrenalectomy: vascular injuries, injuries of the bowel, pleural tears, and injuries to the liver, spleen and pancreas. Fortunately, these complications occur rarely. However, many of these complications can have devastating consequences. Therefore, it's the surgeon's obligation to be aware of the possible complications he might encounter during laparoscopic adrenalectomy. This awareness is essential for their prevention and it helps the laparoscopic surgeon to identify complications intraoperatively.


2D and 3D nanostructuring strategies for thermoelectric materials.
Thermoelectric materials have attracted increased research attention as the implementation of various nanostructures has potential to improve both their performance and applicability. A traditional limitation of thermoelectric performance in bulk materials is the interconnected nature of the individual parameters (for example, it is difficult to decrease thermal conductivity while maintaining electrical conductivity), but through the rational design of nanoscale structures, it is possible to decouple these relationships and greatly enhance the performance. For 2D strategies, newly investigated materials such as graphene, transition metal dichalcogenides, black phosphorus, etc. are attractive thanks to not only their unique thermoelectric properties, but also potential advantages in ease of processing, flexibility, and lack of rare or toxic constituent elements. For 3D strategies, the use of induced porosity, assembly of various nanostructures, and nanoscale lithography all offer specific advantages over bulk materials of the same chemical composition, most notably decreased thermal conductivity due to phonon scattering and enhanced Seebeck coefficient due to energy filtering. In this review, a general summary of the popular techniques and strategies for 2D and 3D thermoelectric materials will be provided, along with suggestions for future research directions based on the observed trends.


Histopathology of salivary glands.
Salivary gland (SG) biopsy is a technique broadly applied for the diagnosis of primary Sjögren's syndrome (pSS), lymphoma accompanying SS, sarcoidosis, amyloidosis, and IgG4-related disease The most peculiar feature of pSS on biopsy is focal lymphocytic sialadenitis. In the past, several histological scores have been reported in the literature to describe glandular involvement during pSS. However, the variability among centres in reporting glandular scores is one of the rationales behind the development of standardised consensus guidance. SGs as well as lacrimal glands are involved in up to 50% of patients with IgG4-related disease with 3 histopathological hallmarks such as dense lymphoplasmacytic infiltration, storiform fibrosis and obliterative phlebitis. SGs can be also affected by amyloidosis with MSG biopsy being more sensitive than that of rectal mucosa or subcutaneous fat. SG involvement is a rare manifestation during sarcoidosis, and the presence of non-caseating granulomas needs to be differentiated from granulomas of other etiology. This review article provides an overview of normal and pathological SGs in the context of rheumatic diseases, identifying key elements in the tissue as diagnostic and prognostic biomarkers, useful in the current clinical practice.


Opportunities and obstacles to screening pregnant women for intimate partner violence during antenatal care in Zimbabwe.
Pregnancy offers an opportunity for midwives to recognise and respond to women experiencing intimate partner violence (IPV). However, most antenatal care interventions have been conducted in private specialist services in high-income countries and do not address the structural and cultural realities of developing country settings. We report on an exploratory qualitative study conducted in antenatal public health facilities in Harare, Zimbabwe, involving six in-depth interviews with midwives and seven FGDs with 64 pregnant and postpartum women. Recorded interviews were transcribed verbatim and analysed using thematic content analysis. We found that identifying and responding to IPV in antenatal care is hampered by inadequate human, financial and infrastructural resources as well as poor support of gender-based violence training for midwives. Midwives had divergent views of their role, with some perceiving IPV as a non-clinical, social and domestic problem that does not require their attention, while others who had been sensitised to the problem felt that it could easily overwhelm them. A comprehensive response to IPV by midwives would be difficult to achieve in this setting but sensitised midwives could respond to cues to violence and ultimately assist abused women in culturally sensitive and appropriate ways.


Quantification and recognition of parkinsonian gait from monocular video imaging using kernel-based principal component analysis.
The computer-aided identification of specific gait patterns is an important issue in the assessment of Parkinson's disease (PD). In this study, a computer vision-based gait analysis approach is developed to assist the clinical assessments of PD with kernel-based principal component analysis (KPCA). Twelve PD patients and twelve healthy adults with no neurological history or motor disorders within the past six months were recruited and separated according to their "Non-PD", "Drug-On", and "Drug-Off" states. The participants were asked to wear light-colored clothing and perform three walking trials through a corridor decorated with a navy curtain at their natural pace. The participants' gait performance during the steady-state walking period was captured by a digital camera for gait analysis. The collected walking image frames were then transformed into binary silhouettes for noise reduction and compression. Using the developed KPCA-based method, the features within the binary silhouettes can be extracted to quantitatively determine the gait cycle time, stride length, walking velocity, and cadence. The KPCA-based method uses a feature-extraction approach, which was verified to be more effective than traditional image area and principal component analysis (PCA) approaches in classifying "Non-PD" controls and "Drug-Off/On" PD patients. Encouragingly, this method has a high accuracy rate, 80.51%, for recognizing different gaits. Quantitative gait parameters are obtained, and the power spectrums of the patients' gaits are analyzed. We show that that the slow and irregular actions of PD patients during walking tend to transfer some of the power from the main lobe frequency to a lower frequency band. Our results indicate the feasibility of using gait performance to evaluate the motor function of patients with PD. This KPCA-based method requires only a digital camera and a decorated corridor setup. The ease of use and installation of the current method provides clinicians and researchers a low cost solution to monitor the progression of and the treatment to PD. In summary, the proposed method provides an alternative to perform gait analysis for patients with PD.


Mulberry Fruit Extract Promotes Serum HDL-Cholesterol Levels and Suppresses Hepatic microRNA-33 Expression in Rats Fed High Cholesterol/Cholic Acid Diet.
Serum high-density lipoprotein cholesterol (HDL-C) levels and cholesterol excretion are closely associated with the risk of cardiovascular complications. The specific aim of the present study was to investigate the cholesterol lowering effect of mulberry fruit in rats fed a high cholesterol/cholic acid diet. Four-week supplementation with mulberry fruit extract significantly decreased serum and hepatic cholesterol (TC), serum low-density lipoprotein cholesterol (LDL-C), and fecal bile acid levels without changes in body weight and food intake (p < 0.05). Mulberry fruit extract significantly inhibited hepatic sterol-regulatory element binding protein (Srebp) 2 gene expression and upregulated hepatic mRNA levels of liver X receptor alpha (Lxr-α), ATP-binding cassette transporter 5 (Abcg5), and cholesterol 7 alpha-hydroxylase (Cyp7a1), which are involved in hepatic bile acid synthesis and cholesterol metabolism (p < 0.05). In addition, hepatic microRNA-33 expression was significantly inhibited by supplementation of mulberry fruit extract (p < 0.05). These results suggest the involvement of miR-33, its associated hepatic bile acid synthesis, HDL formation, and cholesterol metabolism in mulberry fruit-mediated beneficial effects on serum and hepatic lipid abnormalities.


Improved post-transplant survival in the United States for patients with cholangiocarcinoma after 2000.
The incidence of cholangiocarcinoma (CCA) continues to rise. Orthotopic liver transplantation (OLT) can be used for selected patients with localized but unresectable hilar CCA. Although initial post-OLT survival rates were poor, outcomes after introduction of the Mayo Clinic protocol have been more promising and there has been increased interest in OLT for CCA nationally. The aim of this study is to determine post-transplant survival and prognostic factors for patients undergoing OLT for CCA. A retrospective analysis of all patients with CCA listed nationwide for OLT between October 1987 and May 2008 was performed using the Scientific Registry of Transplant Recipients database. Survival curves were generated using the Kaplan-Meier method and compared using log-rank test. Of 595 patients with CCA listed for OLT, 359 (60.3 %) underwent OLT. Median age at OLT was 49 years, 66 % were male and 91 % were Caucasian. The median follow-up time was 2 years. There has been an increasing number of liver transplants performed for CCA since 2000. The 1- and 5-year probability of survival was 85.8 and 51.4 %, respectively. On multivariate analysis, significant prognostic factors for decreased post-OLT survival included transplant before 2000 (HR 11.25, 95 % CI 1.28-98.7) and acute cellular rejection (HR 5.64, 95 % CI 1.14-27.8). Survival after transplant for CCA has improved over time, and OLT is being used more frequently in the treatment of CCA. Significant predictors of post-OLT survival include a history of acute rejection and date of transplant in relation to the publication of Mayo protocol results.


An imprinted crystalline colloidal array chemical-sensing material for detection of trace diethylstilbestrol.
Based on a combination of the molecular imprinting technique and polymerized crystalline colloidal array, we have developed an imprinted crystalline colloidal array (ICCA) chemical-sensing material for the real-time and label-free detection of diethylstilbestrol (DES) in aqueous solution. This novel sensing material was prepared by a noncovalent and self-assembly approach using liquid monodispersed DES-imprinted colloidal spheres and was characterized by a three-dimensional (3D) ordered opal structure in which numerous nanocavities were derived from DES imprinting. Thus, the inherent high affinity of the nanocavities allowed ICCA to recognize DES with high specificity, and changes of the ordered periodic structure enabled ICCA to transfer the recognition events into readable optical signals (label-free). Owing to the special opal structure and without interference from the bulk hydrogel film, the ICCA enabled the rapid and sensitive detection of the target analyte. The understanding of the recognizing response has also been advanced by using molecular modeling software to compute rational interaction between the template molecules and the function monomers. After careful optimization of the assay conditions, the ICCA could decrease its diffraction intensity within just 7 min according to the DES concentration from 2 ng mL(-1) to 8.192 μg mL(-1), whereas there were no obvious diffraction intensity changes for the DES analogues. The adsorption results showed that the homogenous structure and large surface area of ICCA could improve its adsorption capacity. Therefore, such a sensing material with high selectivity, high sensitivity, high stability, and easy operation might offer an attractive alternative for establishing optical sensors for the rapid real-time monitoring of different residues in food and the environment.


Radiative transfer two-stream shape factors for ocean optics.
The mean upward-scattering coefficient of the downward-traveling photons and the mean downward-scattering coefficient of the upward-traveling photons are two factors needed for the two-stream approximation to the radiative-transfer equation. Numerical values of each shape factor just beneath the surface and at asymptotic depths give an indication of the range of values at intermediate depths in spatially uniform waters with no sources and are used to obtain an approximate depth-dependent model for each shape factor. The shape factors are computed for different surface-illumination conditions, wavelengths, and chlorophyll concentrations.


Progressive Tau Accumulation in Alzheimer Disease: 2-Year Follow-up Study.
Tau PET enables in vivo visualization and quantitation of tau accumulation in Alzheimer disease (AD). In cross-sectional tau PET studies, tau burden reflects disease severity and phenotypic variation. We investigated longitudinal changes in cortical tau accumulation and their association with cognitive decline in patients with AD. Methods: We enrolled 107 participants (45 amyloid-β-negative cognitively unimpaired [CU-], 7 amyloid-β-positive cognitively unimpaired [CU+], 31 with prodromal AD [mild cognitive impairment; MCI+], and 24 with AD dementia [DEM+]) who completed 2 baseline PET scans (18F-flortaucipir and 18F-florbetaben), MRI, and neuropsychologic tests. All participants underwent the same assessments after 2 y. After correcting for partial-volume effect, we created SUV ratio (SUVR) images. By using a linear mixed-effect model, we investigated the changes in SUVR across time within each group. We also investigated a correlation between the progression of tau accumulation and cognitive decline. Results: In contrast to no change in global cortical SUVR in the CU- and CU+ groups during the 2-y period, global cortical SUVR increased by 0.06 (2.9%) in the MCI+ group and 0.19 (8.0%) in the DEM+ group at follow-up. The MCI+ group was associated with additional tau accumulation predominantly in the medial and inferior temporal cortices, whereas the DEM+ group showed increases in the lateral temporal cortex. Progressive tau accumulation occurred in the diffuse cortical areas in the MCI+ patients who developed dementia and the DEM+ patients who showed deterioration of global cognition, whereas there was only a small increase of additional tau accumulation in the lateral temporal cortex in those who did not show worsening of cognition. Deterioration of global cognition and language functions was associated with progression of diffuse tau accumulation in the association neocortex. Conclusion: Progressive tau accumulation occurs in prodromal AD and DEM patients in the cortical areas at different levels of tau accumulation. Progression of cognitive dysfunction may be related to the additional tau accumulation in regions of higher Braak stage. 18F-flortaucipir PET is an imaging biomarker for monitoring the progression of AD.


Taxa: An R package implementing data standards and methods for taxonomic data.
The taxa R package provides a set of tools for defining and manipulating taxonomic data. The recent and widespread application of DNA sequencing to community composition studies is making large data sets with taxonomic information commonplace. However, compared to typical tabular data, this information is encoded in many different ways and the hierarchical nature of taxonomic classifications makes it difficult to work with. There are many R packages that use taxonomic data to varying degrees but there is currently no cross-package standard for how this information is encoded and manipulated. We developed the R package taxa to provide a robust and flexible solution to storing and manipulating taxonomic data in R and any application-specific information associated with it. Taxa provides parsers that can read common sources of taxonomic information (taxon IDs, sequence IDs, taxon names, and classifications) from nearly any format while preserving associated data. Once parsed, the taxonomic data and any associated data can be manipulated using a cohesive set of functions modeled after the popular R package dplyr. These functions take into account the hierarchical nature of taxa and can modify the taxonomy or associated data in such a way that both are kept in sync. Taxa is currently being used by the metacoder and taxize packages, which provide broadly useful functionality that we hope will speed adoption by users and developers.


Gamma-secretase activity of presenilin 1 regulates acetylcholine muscarinic receptor-mediated signal transduction.
Familial Alzheimer's disease (FAD) presenilin 1 (PS1) mutations give enhanced calcium responses upon different stimuli, attenuated capacitative calcium entry, an increased sensitivity of cells to undergo apoptosis, and increased gamma-secretase activity. We previously showed that the FAD mutation causing an exon 9 deletion in PS1 results in enhanced basal phospholipase C (PLC) activity (Cedazo-Minguez, A., Popescu, B. O., Ankarcrona, M., Nishimura, T., and Cowburn, R. F. (2002) J. Biol. Chem. 277, 36646-36655). To further elucidate the mechanisms by which PS1 interferes with PLC-calcium signaling, we studied the effect of two other FAD PS1 mutants (M146V and L250S) and two dominant negative PS1 mutants (D257A and D385N) on basal and carbachol-stimulated phosphoinositide (PI) hydrolysis and intracellular calcium concentrations ([Ca2+]i) in SH-SY5Y neuroblastoma cells. We found a significant increase in basal PI hydrolysis in PS1 M146V cells but not in PS1 L250S cells. Both PS1 M146V and PS1 L250S cells showed a significant increase in carbachol-induced [Ca2+]i as compared with nontransfected or wild type PS1 transfected cells. The elevated carbachol-induced [Ca2+]i signals were reversed by the PLC inhibitor neomycin, the ryanodine receptor antagonist dantrolene, the general aspartyl protease inhibitor pepstatin A, and the specific gamma-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester. The cells expressing either PS1 D257A or PS1 D385N had attenuated carbachol-stimulated PI hydrolysis and [Ca2+]i responses. In nontransfected or PS1 wild type transfected cells, N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester and pepstatin A also attenuated both carbachol-stimulated PI hydrolysis and [Ca2+]i responses to levels found in PS1 D257A or PS1 D385N dominant negative cells. Our findings suggest that PS1 can regulate PLC activity and that this function is gamma-secretase activity-dependent.


Stability of blood-based biomarkers of Alzheimer's disease over multiple freeze-thaw cycles.
Freeze-thaw instability may contribute to preanalytical variation in blood-based biomarker studies. We investigated the effects of up to four freeze-thaw cycles on single molecule array immunoassays of serum neurofilament light chain and plasma total tau, amyloid β 1-40 (Aß40), and Aβ 1-42 (Aβ42). Individuals who had peripheral venepuncture during investigation of suspected neurodegenerative disease were recruited. After standardized preprocessing, 200 μL of plasma and serum aliquots were stored at -80°C within 60 minutes. Aliquots underwent one to four freeze-thaw cycles. There was no significant difference across four freeze-thaw cycles for serum neurofilament light chain (n = 12), plasma total tau (n = 11), or plasma Aβ42 (n = 12). For plasma Aβ40 (n = 14), there were significant median reductions by ratios of .96 and .92 at the third and fourth cycles, respectively. Up to four freeze-thaw cycles do not influence single molecule array blood biomarkers of neurofilament light chain, total tau, or Aβ42, with at most minor reductions in Aβ40.


Plasma protein binding of flurbiprofen: enantioselectivity and influence of pathophysiological status.
Ultrafiltration was used to study the enantioselective binding of R-, S- and racemic flurbiprofen, an arylpropionic acid nonsteroidal anti-inflammatory drug, to human plasma protein in eight normal volunteers. The percent free of the R was greater than that of the S for most subjects, but the ratio of percent unbound R to S ranged from 0.52 to 1.33. In extensive studies with a single representative plasma, the percent unbound of the individual enantiomers was independent of concentration up to 26 micrograms/ml, although the binding of the S enantiomer (0.048 +/- 0.007%) was greater than that of the R (0.082 +/- 0.017%), whereas that of the racemate (0.065 +/- 0.028%) was intermediate. At supratherapeutic concentrations of the enantiomers, the percent free of both R- and S-flurbiprofen increased to 0.252 at 100 micrograms/ml. A one-site binding model with nonspecific binding described the data well for R-, S- and racemic flurbiprofen; resulting Ka values were 3.39 X 10(6), 7.31 X 10(6) and 4.35 X 10(6) M-1, respectively. The binding of each enantiomer in the presence of varying amounts of its antipode was described best by a simple competitive inhibition model where mutual displacement occurred as concentrations increased beyond 26 micrograms/ml. The binding of racemic flurbiprofen in elderly and obese volunteers and patients with liver disease was not significantly different from normal subjects, but binding was less in hypoalbuminemic patients and patients with renal impairment. Uremia decreased the binding of R-flurbiprofen preferentially.


Photobiomodulation for Global Cerebral Ischemia: Targeting Mitochondrial Dynamics and Functions.
Hypothermia is currently the only approved therapy for global cerebral ischemia (GCI) after cardiac arrest; however, it unfortunately has multiple adverse effects. As a noninvasive procedure, photobiomodulation (PBM) therapy has emerged as a potential novel treatment for brain injury. PBM involves the use of low-level laser light therapy to influence cell behavior. In this study, we evaluated the therapeutic effects of PBM treatment with an 808-nm diode laser initiated 6 h after GCI. It was noted that PBM dose-dependently protected against GCI-induced neuronal death in the vulnerable hippocampal CA1 subregion. Functional assessments demonstrated that PBM markedly preserved both short-term (a week) and long-term (6 months) spatial learning and memory function following GCI. Further mechanistic studies revealed that PBM post-treatment (a) preserved healthy mitochondrial dynamics and suppressed substantial mitochondrial fragmentation of CA1 neurons, by reducing the detrimental Drp1 GTPase activity and its interactions with adaptor proteins Mff and Fis1 and by balancing mitochondrial targeting fission and fusion protein levels; (b) reduced mitochondrial oxidative damage and excessive mitophagy and restored mitochondrial overall health status and preserved mitochondrial function; and (c) suppressed mitochondria-dependent apoptosome formation/caspase-3/9 apoptosis-processing activities. Additionally, we validated, in an in vitro ischemia model, that cytochrome c oxidase served as a key PBM target for mitochondrial function preservation and neuroprotection. Our findings suggest that PBM serves as a promising therapeutic strategy for the functional recovery after GCI, with mechanisms involving PBM's preservation on mitochondrial dynamics and functions and the inhibition of delayed apoptotic neuronal death in GCI.