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Giant cell arteritis (GCA) is a systemic inflammatory vasculitis that affects medium- and large-sized arteries and can result in permanent vision loss. In rare instances, Horner syndrome has been noticed at the time of GCA diagnosis, although the mechanism of both diagnoses occurring at the same time is not entirely understood. We reviewed 53 charts of all patients diagnosed with biopsy-proven GCA in tertiary neuro-ophthalmology practice to find patients who presented with new onset of Horner syndrome at the time of GCA diagnosis.Two patients with biopsy-confirmed GCA who presented with concurrent Horner syndrome were found. Data on age, sex, and ophthalmic and neuroradiologic examination findings were collected.Patient 1 was a 67-year-old man who presented with new onset of vertical binocular diplopia and was diagnosed with right fourth cranial nerve palsy. He then developed left ptosis and miosis, and was diagnosed with Horner syndrome by pharmacologic testing. He also had persistently elevated inflammatory markers. Patient 2 was a 71-year-old man who presented with new onset of binocular vertical diplopia, bitemporal headaches, and jaw ache. Both of his inflammatory markers were elevated. On examination, he had left ptosis and myosis, and small comitant left hypertropia. The diagnosis of left Horner syndrome was confirmed on pharmacologic testing and left hypertropia was attributed to skew deviation. Both patients underwent temporal artery biopsy, which confirmed the diagnosis of GCA. Treatment with high dose of oral corticosteroids commenced, and vertical diplopia has completely resolved in both patients. Horner syndrome persisted in Patient 1 and resolved in Patient 2. MRI and MR angiography of the brain and neck were unrevealing in both patients.This case series describes 2 patients with new diagnosis of GCA and concurrent Horner syndrome, with new diagnosis of likely nuclear/fascicular fourth nerve palsy in one patient and skew deviation in the other. In both patients, vasculitis presumptively affected vertebral arteries and their branches supplying the first-order sympathetic neurons in the brainstem. Considering the severe complication of permanent vision loss in GCA, this diagnosis should be considered in older patients presenting with concurrent new onset of Horner syndrome.

Play behaviors are generally considered to be beneficial and necessary for young, developing animals. Expression of play, however, is sensitive to several post-natal factors including maternal care. In this study, we observed whether maternal stress and maternal care behaviors were related with adolescent play behavior in the degu (Octodon degus), a diurnal rodent species native to central Chile. By observing pairs of mothers that were either unstressed (habituated to captivity) or stressed (only recently introduced into captivity), the frequencies of both social and locomotor play behaviors in the offspring, along with general behaviors and time of rest, were scored from 2 to 20 days after parturition. All scored play behaviors increased in frequency over time, indicating that degu pups become more active and playful as they mature. Additionally, we also found that pups from unstressed mothers expressed higher frequencies of locomotor play behavior (Running and Frisky Hops) and some social affiliation behaviors (Nose-to-Nose Contact) compared to pups from stressed mothers. Furthermore, these behaviors were significantly and positively related with the amount of maternal care received. Our results suggest that development of certain play behaviors are affected by maternal stress, likely via alterations in maternal care.

Cdk8 of the RNA polymerase II mediator kinase complex regulates gene expression by phosphorylating sequence-specific transcription factors. This function is conserved amongst eukaryotes, but the signals and mechanisms regulating Cdk8 activity and phosphorylation of its substrates are unknown. Full induction of the GAL genes in yeast requires phosphorylation of the transcriptional activator Gal4 by Cdk8. We used a screen to identify regulators of the Cdk8-dependent phosphorylation on Gal4, from which we identified multiple mutants with defects in TORC1 signaling. One mutant, designated gal four throttle 1 (gft1) was identified as a recessive allele of hom3, encoding aspartokinase, and mutations in hom3 caused effects typical of inhibition of TORC1, including rapamycin sensitivity and enhanced nuclear localization of the TORC1-responsive transcription factor Gat1. Mutations in hom3 also inhibit phosphorylation of Gal4 in vivo at the Cdk8-dependent site on Gal4, as did mutations of tor1, but these mutations did not affect activity of Cdk8 assayed in vitro. Disruption of cdc55, encoding a regulatory subunit of the TORC1-regulated protein phosphatase PP2A, suppressed the effect of hom3 and tor1 mutations on GAL expression, and also restored phosphorylation of Gal4 at the Cdk8-dependent site in vivo. These observations demonstrate that TORC1 signaling regulates GAL induction through the activity of PP2A/Cdc55 and suggest that Cdk8-dependent phosphorylation of Gal4 is opposed by PP2A/Cdc55 dephosphorylation. These results provide insight into how induction of transcription by a specific inducer can be modulated by global nutritional signals through regulation of Cdk8-dependent phosphorylation., Horvath, Hawe et al. examine regulators of Cdk8-dependent Gal4 activity in a screen for defects in the LTA response of gal3 yeast. They identify TORC1 components tor1 and tco89 and the downstream PP2A/Cdc55 phosphatase as regulators of Gal4. Additionally, genetic analysis indicates that mutations in hom3, which encodes aspartate kinase, produce defects in TOR signaling. These results demonstrate that regulation of Gal4 by phosphorylation is modulated by the TOR nutritional signaling pathway through the activity of PP2A/Cdc55.

BACKGROUND/OBJECTIVES: No guidelines exist for the investigation of treatable causes of chronic optic neuropathy, including sarcoidosis, lupus, and syphilis. The purpose of this study was to determine the diagnostic yield of screening blood work (ACE (Angiotensin Converting Enzyme) for sarcoidosis, Antinuclear Antibodies (ANA) for lupus, CMIA (chemiluminescence microparticle enzyme immunoassay) for syphilis) and contrast-enhanced MRI brain and orbits in atypical unilateral chronic optic neuropathy. SUBJECTS/METHODS: Retrospective review from February 2012 to June 2018 at a neuro-ophthalmology practice. Six hundred and eighty-three consecutive charts with optic neuropathy were reviewed. Inclusion criteria were unilateral chronic optic neuropathy and a work-up including contrast-enhanced MRI brain and orbits, CBC, ESR, CRP, ANA, CMIA, and ACE. Exclusion criteria were optic nerve swelling in either eye on initial assessment or an established cause of optic neuropathy. The main outcome measure was diagnostic yield. RESULTS: Fifty-seven patients were included. One patient had elevated ACE, seven had positive ANA titers, and three had positive CMIA. Zero patients were diagnosed with sarcoidosis, one patient was diagnosed with lupus-related optic neuropathy, and one patient was diagnosed with syphilitic optic neuropathy. The diagnostic yield of ACE was 0%, ANA was 1.75%, and CMIA was 1.75%. MRI revealed planum sphenoidale meningioma causing compressive optic neuropathy in one patient, giving it a diagnostic yield of 1.82%. CONCLUSION: Routine screening blood work (ACE, ANA, CMIA) and MRI brain and orbits for chronic idiopathic unilateral optic neuropathy has low diagnostic yield, especially if clinical suspicion for syphilis, lupus, and sarcoidosis is low. MRI should still be performed in all cases in order to rule out compressive lesions.

Incubation of craving refers to the intensification of drug-seeking behavior in response to reward-paired cues over the course of abstinence. In rodents, craving and drug-seeking behaviors have been measured by an increase in lever pressing in the absence of reinforcer availability in response to cue presentations. However, craving in rodents is difficult to define and little is known about the behavioral signatures that accompany increased drug-seeking behavior measured by lever pressing. The affective components of relapse are also important, but understudied in rodents. Hormonal fluctuations influence craving for psychostimulants, but little is known about the impact of the estrous cycle on opioid-seeking behavior. This study sought to delineate the behavioral and affective signatures associated with craving, and to examine the influence of the female estrous cycle on craving. Male and female rats underwent 10 d of intravenous opioid self-administration. Separate cohorts of control rats self-administered oral sucrose, a natural nondrug reward. Cue-induced seeking tests were conducted after 1 or 30d of forced abstinence. These sessions were recorded and scored for overall locomotion, instances of sniffing, grooming, or hyperactivity. Ultrasonic vocalizations (USVs) were also recorded to determine affective profiles that accompany opioid seeking. Although active lever presses and overall locomotion increased unanimously over extended abstinence from heroin and sucrose, a sex- and reinforcer-specific behavioral and affective signature of craving emerged. Furthermore, although the female estrous cycle did not affect taking or seeking, it appears to influence more granular behaviors.

Cdk8 of the RNA Polymerase II mediator complex regulates genes by phosphorylating sequence specific transcription factors. Despite conserved importance for eukaryotic transcriptional regulation, the signals regulating Cdk8 are unknown. Full induction of the yeastGALgenes requires phosphorylation of Gal4 by Cdk8, and we exploited this requirement for growth ofgal3yeast on galactose to identify mutants affecting Cdk8 activity. Several mutants from the screen produced defects in TOR signaling. A mutant designatedgalfour throttle (gft) 1,gft1, was identified as an allele ofhom3, encoding aspartokinase. Defects ingft1/hom3caused hypersensitivity to rapamycin, and constitutive nuclear localization of Gat1. Furthermore, mutations oftor1ortco89, encoding TORC1 components, also preventedGALexpression ingal3yeast, andtco89was determined to be allelic togft7. Disruption ofcdc55, encoding a subunit of PP2A regulated by TOR signaling, suppressed the effect ofgft1/hom3, gft7/tco89, andtor1mutations onGALexpression ingal3yeast, but not ofcdk8/srb10disruptions or Gal4 S699A mutation. Mutations ofgft1/hom3andtor1did not affect kinase activity of Cdk8in vitro, but caused loss of Gal4 phosphorylationin vivo. These observations demonstrate that TOR signaling regulatesGALinduction through the activity of PP2A/ Cdc55, and are consistent with the contention that Cdk8-dependent phosphorylation of Gal4 S699 is opposed by PP2A/ Cdc55 dephosphorylation. These results provide insight into how induction of transcription by a specific inducer can be modulated by global nutritional signals through regulation of Cdk8-dependent phosphorylation.

Background Diplopia can be the result of benign or life-threatening etiologies. It is imperative for the emergency physician to be proficient at assessing diplopia and recognize when urgent referral or neuroimaging is required. Objective The first part of this review highlights a simple framework to arrive at the appropriate disposition of diplopic patients presenting to the emergency department (ED). The second part of this review provides more detail and further management strategies. Discussion ED strategies for assessment of diplopia are discussed. Management strategies, such as when to image, what modality of imaging to use, and urgency of referral, are discussed in detail. Conclusions Unenhanced plain computed tomography (CT) of the head or orbits is largely not useful in the work-up of diplopia. Magnetic resonance imaging is preferred for ocular motor nerve palsies. Due to limited resources in the ED, patients with isolated fourth and sixth nerve palsies with the absence of other neurological signs on examination should be referred to Neurology or Ophthalmology for further work-up. All patients presenting with an acute isolated third nerve palsy should be imaged with CT and CT angiography of the brain to rule out a compressive aneurysm. Contrast-enhanced CT imaging of the brain and orbits would be indicated in suspected orbital apex syndrome or a retro-orbital mass, thyroid eye disease, or ocular trauma. CT and CT venogram should be considered in cases of suspected cavernous sinus thrombosis. In any patient over the age of 60 years presenting with recent (1 month) history of diplopia, inflammatory markers should be obtained to rule out giant cell arteritis.

A previously healthy 25-year-old man presented with a 3-weeks history of frontal headache, right-sided ptosis and binocular horizontal diplopia. The diagnosis of right 3rd nerve palsy was made. Magnetic Resonance Imaging/angiography (MRI/A) of the brain was interpreted as normal. Two days later, right facial droop and weakness developed along with lower back pain, paresthesias of both legs and left leg weakness. On exam, he had bilateral upper lid ptosis, bilateral adduction deficits and areflexia of the left patella with bilaterally decreased ankle reflexes. It was concluded that he now has bilateral partial pupil-sparing left 3rd nerve palsies and right peripheral 7th nerve palsy.

Elongator is a ~850 kDa protein complex involved in multiple processes from transcription to tRNA modification. Conserved from yeast to humans, Elongator is assembled from two copies of six unique subunits (Elp1 to Elp6). Despite the wealth of structural data on the individual subunits, the overall architecture and subunit organization of the full Elongator and the molecular mechanisms of how it exerts its multiple activities remain unclear. Using single‐particle electron microscopy (EM), we revealed that yeast Elongator adopts a bilobal architecture and an unexpected asymmetric subunit arrangement resulting from the hexameric Elp456 subassembly anchored to one of the two Elp123 lobes that form the structural scaffold. By integrating the EM data with available subunit crystal structures and restraints generated from cross‐linking coupled to mass spectrometry, we constructed a multiscale molecular model that showed the two Elp3, the main catalytic subunit, are located in two distinct environments. This work provides the first structural insights into Elongator and a framework to understand the molecular basis of its multifunctionality.

Bacteriophages screened and isolated from sewage water samples exhibited antibacterial activities against multi-drug-resistant Escherichia coli strains. Five different water samples from Canadian habitats such as Kamloops Wastewater Treatment Center, Domtar, the Pacific Ocean, Bisaro Anima Cave, and alkali ponds, were used in this study. Four Enterobacteriaceae strains including one non-resistant and three clinical multi-drug Escherichia coli strains (E. coli 15-102, E. coli 15-124, and E. coli 15-318) were selected as target bacteria to screen for the bacteriophages from these collected water samples. Seeded agar assay technique was implemented for the screening. It was found that only sewage water sample exhibited a significant number of plaques count with the E. coli 15-318 (1.82 &times, 102 plaques/plate) cells in comparison to E. coli non-resistant strain K12 (8 plaques/plate). The phage did not produce plaques in the E. coli 15-124 and E. coli 15-102 strains. The bacteriophage, designated EMCL318, was isolated, purified, characterized, and identified to belong to the G4 species of the Family Microviridae, GenBank accession number MG563770. This is an explorative study conducted in order to reveal the viruses as alternative potentials to fight against emerging and existing multi-drug-resistant infectious diseases.

The glycosylated Gag protein (gPr80) of murine leukemia viruses (MLVs) has been shown to exhibit multiple roles in facilitating retrovirus release, infection, and resistance to host-encoded retroviral restriction factors, such as APOBEC3, SERINC3, and SERINC5. One way in which gPr80 helps MLVs to escape host innate immune restriction is by increasing capsid stability, a feature that protects viral replication intermediates from being detected by cytosolic DNA sensors. gPr80 also increases the resistance of MLVs to deamination and restriction by mouse APOBEC3 (mA3). How the gPr80 accessory protein, with its three N -linked glycosylation sites, contributes to these resistance mechanisms is still not fully understood. Here we further characterized the function of gPr80 and, more specifically, revealed that the asparagines targeted for glycosylation in gPr80 also contribute to capsid stability through their parallel involvement in the Pr65 Gag structural polyprotein. In fact, we demonstrate that sensitivity to deamination by the mA3 and human A3 proteins is directly linked to capsid stability. We also show that full-length gPr80 is detected in purified viruses. However, our results suggest that gPr80 is inserted in the N exo C cyto orientation of a type I integral membrane protein. Additionally, our experiments have revealed the existence of a large population of Env-deficient virus-like particles (VLPs) harboring gPr80 inserted in the opposite (N cyto C exo ) polarity, which is typical of type II integral membrane proteins. Overall this study provides new insight into the complex nature of the MLV gPr80 accessory protein. IMPORTANCE Viruses have evolved numerous strategies to infect, spread in, and persist in their hosts. Here we analyze the details of how the MLV-encoded glycosylated Gag (gPr80) protein protects the virus from being restricted by host innate immune defenses. gPr80 is a variant of the structural Pr65 Gag protein with an 88-amino-acid extended leader sequence that directs the protein for translation and glycosylation in the endoplasmic reticulum. This study dissects the specific contributions of gPr80 glycans and capsid stability in helping the virus to infect cells, spread, and counteract the effects of the host intrinsic restriction factor APOBEC3. Overall this study provides further insight into the elusive role of the gPr80 protein.

Edsel B Ing,1 Gabriela Lahaie Luna,2 Andrew Toren,3 Royce Ing,4 John J Chen,5 Nitika Arora,6 Nurhan Torun,7 Otana A Jakpor,8 J Alexander Fraser,9 Felix J Tyndel,10 Arun NE Sundaram,10 Xinyang Liu,11 Cindy TY Lam,1 Vivek Patel,12 Ezekiel Weis,13 David Jordan,14 Steven Gilberg,14 Christian Pagnoux,15 Martin ten Hove21Department of Ophthalmology and Vision Sciences, University of Toronto Medical School, Toronto, 2Department of Ophthalmology, Queen’s University, Kingston, ON, 3Department of Ophthalmology, University of Laval, Quebec, QC, 4Toronto Eyelid, Strabismus and Orbit Surgery Clinic, Toronto, ON, Canada; 5Mayo Clinic, Department of Ophthalmology and Neurology, 6Mayo Clinic, Department of Ophthalmology, Rochester, MN, 7Department of Surgery, Division of Ophthalmology, Harvard Medical School, Boston, MA, 8Harvard Medical School, Boston, MA, USA; 9Department of Clinical Neurological Sciences and Ophthalmology, Western University, London, 10Department of Medicine, University of Toronto Medical School, Toronto, ON, Canada; 11Department of Medicine, Fudan University Shanghai Medical College, Shanghai, People’s Republic of China; 12Roski Eye Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA; 13Departments of Ophthalmology, Universities of Alberta and Calgary, Edmonton and Calgary, AB, 14Department of Ophthalmology, University of Ottawa, Ottawa, ON, 15Vasculitis Clinic, Mount Sinai Hospital, Toronto, ON, CanadaPurpose: To develop and validate a diagnostic prediction model for patients with suspected giant cell arteritis (GCA).Methods: A retrospective review of records of consecutive adult patients undergoing temporal artery biopsy (TABx) for suspected GCA was conducted at seven university centers. The pathologic diagnosis was considered the final diagnosis. The predictor variables were age, gender, new onset headache, clinical temporal artery abnormality, jaw claudication, ischemic vision loss (VL), diplopia, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and platelet level. Multiple imputation was performed for missing data. Logistic regression was used to compare our models with the non-histologic American College of Rheumatology (ACR) GCA classification criteria. Internal validation was performed with 10-fold cross validation and bootstrap techniques. External validation was performed by geographic site.Results: There were 530 complete TABx records: 397 were negative and 133 positive for GCA. Age, jaw claudication, VL, platelets, and log CRP were statistically significant predictors of positive TABx, whereas ESR, gender, headache, and temporal artery abnormality were not. The parsimonious model had a cross-validated bootstrap area under the receiver operating characteristic curve (AUROC) of 0.810 (95% CI =0.766–0.854), geographic external validation AUROC’s in the range of 0.75–0.85, calibration pH–L of 0.812, sensitivity of 43.6%, and specificity of 95.2%, which outperformed the ACR criteria.Conclusion: Our prediction rule with calculator and nomogram aids in the triage of patients with suspected GCA and may decrease the need for TABx in select low-score at-risk subjects. However, misclassification remains a concern.Keywords: temporal artery biopsy, diagnosis, prediction rule, nomogram, giant cell arteritis, validation

Purpose To measure and compare the cytokine concentrations in the aqueous humor of patients with acute nonarteritic anterior ischemic optic neuropathy (NAION) and normal age-related cataract controls. Design Prospective, comparative observational study. Methods Aqueous humor samples were obtained in 10 patients with acute NAION (within 14 days of symptom onset) and 15 control patients with age-related cataract. The levels of 6 cytokines—vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-2, IL-6, and IL-8—were determined using a multiplex bead immunoassay. The clinical characteristics of patients were also collected for correlation analysis. Results The mean concentration of VEGF (94.1 ± 40.4 pg/mL) was significantly higher in the NAION group compared to the cataract controls (52.2 ± 20.8 pg/mL; P = .010) and the mean concentration of IL-2 (5.56 ± 1.27 pg/mL) was significantly lower in the NAION group than in the cataract controls (16.6 ± 14.0 pg/mL; P = .002). There were no differences in the concentration of TNF-α, IL-1β, IL-6, and IL-8. There was a strong negative correlation between the VEGF concentration and the peripapillary retinal nerve fiber layer (RNFL) thickness at presentation (r = −0.657, P = .055). There was no significant correlation between the RNFL thickness and any other cytokines, the mean deviation on 24-2 Humphrey visual fields, or the duration of vision loss. Conclusions Acute NAION is associated with higher VEGF and lower IL-2 concentrations without a change in other inflammatory cytokines. This has implications for future therapeutic interventions and diagnostic testing in patients with this acute optic neuropathy.

Holocarboxylase synthetase (HLCS) deficiency is a rare autosomal recessive disorder that presents with multiple life-threatening metabolic derangements including metabolic acidosis, ketosis, and hyperammonemia. A majority of HLCS deficiency patients respond to biotin therapy; however, some patients show only a partial or no response to biotin therapy. Here, we report a neonatal presentation of HLCS deficiency with partial response to biotin therapy. Sequencing of HLCS showed a novel heterozygous mutation in exon 5, c.996G>C (p.Gln332His), which likely abolishes the normal intron 6 splice donor site. Cytogenetic analysis revealed that the defect of the other allele is a paracentric inversion on chromosome 21 that disrupts HLCS. This case illustrates that in addition to facilitating necessary family testing, a molecular diagnosis can optimize management by providing a better explanation of the enzyme’s underlying defect. It also emphasizes the potential benefit of a karyotype in cases in which molecular genetic testing fails to provide an explanation.

To test whether macroalgae affect microbial colonizers in close proximity in a phylum-specific fashion, the community richness of planktonic bacteria and fungi was analyzed with selective oligonucleotide probes targeting the Cytophaga/Flavobacterium/Bacteroides (CFB), Alphaproteobacteria and Roseobacter group and the ITS1 region of marine fungi. Naturally occuring planktonic microorganisms were incubated in the presence of macroalgae or in seawater previously conditioned with macroalgal metabolites. The red algae Ceramium rubrum and Mastocarpus stellatus as well as seawater conditioned with these algae reduced the community composition of bacteria to a greater extent than the brown alga Laminaria digitata, indicating that metabolites differed among macroalgae or that the susceptibility of planktonic bacteria towards alga-derived antimicrobials correlated with their phylogenetic affiliation. The most affected phylotypes belonged to the CFB and the Roseobacter clade. The planktonic fungal community was only affected in the presence of macroalgae and not in algal-conditioned water, but with a specificity different from that observed for bacteria. The macroalgae L. digitata and M. stellatus exhibited more pronounced antifungal effects than C. rubrum. This study demonstrates macroalgal defenses against epiphytic microorganisms based on natural delivery mechanisms of allelochemicals utilizing a culture-independent approach, thus minimizing the ecological bias inherent to culture-dependent studies based on few microbial isolates.

A variety of macroalgae (Ceramium rubrum, Corallina officinalis, Palmaria palmata, Mastocarpus stellatus, Fucus vesiculosus, Cladophora rupestris, Ulva sp.) were investigated by scanning electron microscopy to visualize epiphytic colonizers. The macroalgae differed in terms of their epiphytic coverage of bacteria, fungi and diatoms. Macroalgae, largely devoid of epiphytic diatoms, were hypothesized to employ effective antifouling means to reduce epiphytic coverage, whilst heavily fouled macroalgae were proposed to lack antifouling strategies. To test these hypotheses from an allelochemical perspective with regard to fouling diatoms, dichloromethane-methanol (1:1) crude extracts of macroalgae were concentrated in dimethylsulfoxide and investigated in diatom attachment and proliferation assays using four benthic diatoms (Nitzschia sp., Navicula phyllepta, Navicula arenaria and Amphora sp.). Algal extracts exhibited a distinct pattern of activity against the test diatoms, suggesting a targeted and selective effect of macroalgal metabolites on individual fouling diatoms. The main outcome of this study was that visual inspection and quantitative categorization of epiphytic colonizers on macroalgal thalli could not be used to predict reliably whether macroalgae employed a chemical defense mechanism.

Many macroalgae employ chemical means to suppress epibiosis by micro- and macroorganisms. Previous studies have focused on the effects of tissue extracts of entire algae on a few bacterial isolates, thereby missing not only ecologically relevant bacteria but also natural delivery mechanisms of algal antimicrobial agents. In this study, we investigated the potential antimicrobial effects of waterborne macroalgal metabolites utilizing a culture-independent approach to compare the bacterial community richness in seawater in the presence and absence of macroalgae. The methodology comprised the collection of planktonic bacteria in algal culture water on membrane filters followed by filter PCR and denaturant gradient gel electrophoresis (DGGE) of 16S rRNA gene sequences of harvested bacteria with universal primers. Similarity analysis distinguished two groups of macroalgae under investigation, one of which showed >55% difference, and the other

Extracellular polymers (EPS) produced by diatoms have been demonstrated as potent mediators of larval settlement in Hydroides elegans. Few studies have addressed the importance of environmental parameters (e.g. temperature and salinity) on quantity and quality of diatom EPS as larval settlement cues. The growth rates of the benthic pennate diatoms Achnanthes sp. and Nitzschia constricta reached a maximum and a minimum level at high and low temperature and salinity, respectively. A positive correlation was observed between the growth rate of diatoms and their production of EPS. In order to decouple the quantitative effect of diatom films from the potential qualitative effect of diatom EPS on larval settlement, the same amount of diatom EPS obtained from different growth conditions were immobilized in hydrogels and investigated in larval settlement assays. The diatom EPS obtained from high temperature treatments were more inductive to larval settlement than the low-temperature treatments, irrespective of the salinity setting. The qualitative differences of the diatom EPS samples under investigation were further analyzed by bioassay-guided gel-chromatography. The bioactive EPS fraction of Achnanthes sp. mainly consisted of large macro- molecules, while the bioactive EPS fraction of N. constricta consisted of both large and small macro- molecules. Due to their sensitivity to changes in environmental parameters, diatoms may serve as small scale proxies of substratum suitability for larval settlement of benthic marine invertebrates, analogous to the reported role of bacteria in marine biofilms.

Larval settlement in the polychaete Hydroides elegans is mediated by marine biofilms, which are complex agglomerates of bacteria, diatoms, fungi and protozoa. The induction of H. ele- gans larval settlement by marine biofilms has been mainly attributed to bacteria and diatoms. In con- trast to bacteria-derived settlement cues, the nature and origin of diatom-derived settlement cues is poorly understood. In this study, we present the first investigation on the nature and origin of larval settlement cues produced by marine diatoms. Diatoms with inductive (i.e. Achnanthes sp. and Nitzschia constricta) and non-inductive (i.e. Amphora tenerrima and N. frustulum) effects on larval settlement of H. elegans were selected as model strains in this investigation. Larval settlement bio- assays with a choice between monospecies diatom films and unfilmed substratum revealed that the diatom-derived settlement cue was water-insoluble and associated with the film surface. There was a clear correlation between the surface coverage of diatoms in films and their inductive effect on larval settlement. In the case of Achnanthes sp., even the lowest surface coverage of 1.8% induced larval settlement significantly more than the control of filtered seawater. The interstitial distance between diatom cells at this low film density was within the body size range of H. elegans larvae (100 ± 30 µm). The inductive effect of diatom films on larval settlement prevailed even after heat treatments (121°C for 1 h) that completely killed the diatoms, as verified with the vital stain fluores- cein diacetate. These results suggest that the induction of larval settlement by diatoms is not related to their viability, and that, contrary to marine bacteria, diatom-induced larval settlement cues are composed of heat-stable surface components, such as capsular extracellular polysaccharides. These results not only demonstrate that different components of biofilms play inductive and inhibitive roles on larval settlement, but also that their relative space occupation deserves consideration regarding their roles as mediators of larval settlement.

Larval settlement in the fouling polychaete Hydroides elegans Haswell (1883) is mediated by natural biofilms, a complex consortium of marine bacteria, benthic diatoms, protozoa, fungi and adsorbed organic matter. Currently, the inductive effect of natural biofilms on larval settlement is attributed to the bacterial component. In this paper, we present results of an investigation of the diatom component of natural biofilms with respect to induction of larval settlement in H. elegans. Thirty-two diatom strains were isolated from natural biofilms, identified and individually investigated in still-water laboratory settlement assays. A multiple pairwise comparison of larval settlement rates grouped the diatoms into 3 statistical categories: (1) 'inductive'; same percentage of larval settlement as in the positive control (glass slide with natural biofilm); (2) 'no effect'; same percentage of larval settlement as in the negative control (clean glass slide); and (3) 'weakly inductive'; percentage of larval settlement significantly higher and lower than in the negative and the positive control, respectively. Out of 32 diatom strains, 3 (10 %) fell into the first, 19 (59%) into the second and 10 (31%) into the third category. The variability in larval settlement was not correlated with the density of diatoms in the monospecific films. Despite sterile measures and the use of antibiotics, diatom films under investigation were not axenic, with bacterial abundances ranging from 200 to 8000 cells mm -2 . No quantitative correlation between the bacterial abundance in diatom films and the percentage of larval settlement was observed. To evaluate the qualitative impact of the bacterial fraction in the mixed diatom-bacteria films on the induction of larval settlement, 6 representative diatom strains, comprising the 3 categories stated above, were selected to create diatom-free microbial films with the bacterial contaminants of each culture. Whilst the bacterial abundance in the non-axenic diatom films and the diatom-free control films was statistically the same, the percentage of larval settlement towards these treatments differed significantly for the diatom strains belonging to category (1) and (3). These categories comprised 4 different genera of diatoms, indicating that the observed inductive effect on larval settlement was not restricted to a certain diatom genus. Our results suggest that in addition to microbial films, benthic diatoms may significantly contribute to the mediation of larval settlement behaviour in H. elegans by marine biofilms.

The frequent occurrence of persistent or relapsed disease after induction chemotherapy in AML necessitates a better understanding of the clonal relationship of AML in various disease phases. In this study, we used SNP 6.0 array-based genomic profiling of acquired copy number aberrations (aCNA) and copy neutral LOH (cnLOH) together with sequence analysis of recurrently mutated genes to characterize paired AML genomes. We analyzed 28 AML sample pairs from patients who achieved complete remission with chemotherapy and subsequently relapsed and 11 sample pairs from patients with persistent disease after induction chemotherapy. Through review of aCNA/cnLOH and gene mutation profiles in informative cases, we demonstrate that relapsed AML invariably represents re-emergence or evolution of a founder clone. Furthermore, all individual aCNA or cnLOH detected at presentation persisted at relapse indicating that this lesion type is proximally involved in AML evolution. Analysis of informative paired persistent AML disease samples uncovered cases with 2 coexisting dominant clones of which at least one was chemotherapy sensitive and one resistant, respectively. These data support the conclusion that incomplete eradication of AML founder clones rather than stochastic emergence of fully unrelated novel clones underlies AML relapse and persistence with direct implications for clinical AML research.

We identified a novel 6.33 Mb deletion of 1q21.3q23.3 (hg18; chr1: 153035245-159367106) in two siblings presenting with blepharophimosis, ptosis, microbrachycephaly, severe psychomotor, and intellectual disability. Additional common features include small corpus callosum, normal birth length and head circumference, postnatal growth restriction, low anterior hairline, upturned nose, bilateral preauricular pits, widely spaced teeth, gingival hypertrophy, left ventricular dilatation with decreased biventricular systolic function, delayed bone age, 5th finger clinodactyly, short 3rd digit, hyperconvex nails, obstructive and central sleep apnea, and bilateral heel contractures. Fluorescence in situ hybridization (FISH) performed in the mother of both children showed an apparently balanced, intrachromosomal insertional translocation of 1q21.3q23.3 to 1q42.12. The sibling recurrence likely arose by a maternal meiotic crossing over on the rearranged chromosome 1 between the deleted region and the insertion. We hypothesize that the decreased cardiac function and contractures may be related to LMNA haploinsufficiency. This case illustrates the importance of FISH when attempting to determine inheritance of a copy-number variation and emphasize the value of evaluating known haploinsufficiency phenotypes for genes in deleted regions.

This chapter first describes the role of Traditional Chinese Medicine (TCM) in health care. It then reviews the literature on the effectiveness of TCM with a special focus on primary care. An appraisal of the outcome measures in the context of TCM is made. The relationship between TCM and the concept of health-related quality of life (HRQOL) is discussed. The current applications and limitations of the HRQOL measures derived from Western culture to TCM are identified. The chapter ends with an overview of Chinese culture specific measures for evaluating the effectiveness of TCM in primary care.

Polybrominated diphenylethers (PBDEs) are found ubiquitously in marine environments worldwide. Sediment is the major sink of PBDEs, with the congener BDE 47 being most abundant. In this study, laboratory experiments were carried out to test the hypothesis that contamination of BDE 47 at environmentally realistic sediment concentrations can alter polychaete larval settlement. Using multiple-choice experiment, settlement of three polychaete species (Pseudopolydora vexillosa, Polydora cornuta, and Capitella sp. I) on four types of spiked sediment was studied and compared: (i) low BDE 47 concentration (0.5 ng g(-1) dry weight); (ii) high BDE 47 concentration (3.0 ng g(-1) dry weight), (iii) hexane (solvent control), and (iv) natural sediment (control). Our results showed that settlement of P. vexillosa and Capitella sp. I larvae was significantly promoted, while settlement of P. cornuta reduced, at high BDE 47 concentration in sediment compared with the respective controls under both short- (24 h) and long-term (4 week) exposures. After 4 weeks, body burden of BDE 47 in all polychaete species was directly related to the spike concentration, and body length of settled juveniles of P. vexillosa and Capitella sp. I at the high-concentration treatment was significantly longer compared with that of other treatments and controls. For the first time, we demonstrated that environmentally realistic concentrations of BDE 47 in sediment can affect polychaete settlement in species-specific and dose-dependent manners. Given the global contamination of PBDE in marine sediment, BDE 47 may potentially alter the settlement pattern of marine polychaetes and hence their benthic composition over large areas.

Summary Background Traditional Chinese medicine (TCM) plays an important role in the primary care system in many places, but research evidence on its effectiveness is largely lacking. The aim of the present study was to compare the effectiveness between TCM and Western medicine (WM) consultations in primary care. Objectives To evaluate whether medical consultations could improve the quality of life and health condition of patients in primary care and to find out whether there was any difference in the effectiveness bewteen TCM and WM. Design, setting and subjects This was a prospective, longitudinal study on 290 patients of one TCM public and 841 patients of two WM general outpatient clinics (GOPC) in Hong Kong when they consulted for an episodic illness. Methods All patients attending a TCM GOPC in TWH, and the two WM GOPC (TWH and ALC), who fullfilled the inclusion criteria were invited to participate. Each patient answered a structured questionnaire on the presenting complaint, socio-demography, chronic morbidity and service utilization, the Chinese Quality of Life instrument (ChQOL) and the SF-36V2 Health Survey immediately before and two weeks after the doctor consultation. The Global Rating on change Scale (GRS) was also administered in the week 2 assessment. Outcome measures The primary outcomes were changes in the ChQOL and SF-36V2 HRQOL scores. Secondary outcomes included the GRS score. The significance of the change within individual were tested by paired t -tests. The differences in change in scores between WM and TCM were tested by independent sample- t -tests or chi-square, as appropriate. Multivariate regresions were used to determine the independent effect of type of medicine on the change in HRQOL scores. Results Mean ChQOL and SF-36V2 scores of subjects improved significantly two weeks after TCM or WM consultations in all domains except for the Physical form domain of ChQOL. The greatest improvements were found in the SF-36V2 physical-health related domains. 78% TCM clinics and 71% of subjects WM clinics reported an improvement in GRS. The proportion of subjects who had improvement in HRQOL scores were lower among subjects consulting the WM clinic (72.3%) than those consulting TCM clinics (100%) but the difference was not significant after correction for baseline scores. Conclusions Both TCM and WM consultations were associated with significant improvement in HRQOL in over 90% of patients. There was no singificant difference between the effectiveness of TCM and WM consultations. The results support the role of TCM as an alternative primary care service in Hong Kong.

To test whether macroalgae affect microbial colonizers in close proximity in a phylum-specific fashion, the community richness of planktonic bacteria and fungi was analyzed with selective oligonucleotide probes targeting the Cytophaga/Flavobacterium/Bacteroides (CFB), Alphaproteobacteria and Roseobacter group and the ITS1 region of marine fungi. Naturally occuring planktonic microorganisms were incubated in the presence of macroalgae or in seawater previously conditioned with macroalgal metabolites. The red algae Ceramium rubrum and Mastocarpus stellatus as well as seawater conditioned with these algae reduced the community composition of bacteria to a greater extent than the brown alga Laminaria digitata, indicating that metabolites differed among macroalgae or that the susceptibility of planktonic bacteria towards alga-derived antimicrobials correlated with their phylogenetic affiliation. The most affected phylotypes belonged to the CFB and the Roseobacter clade. The planktonic fungal community was only affected in the presence of macroalgae and not in algal-conditioned water, but with a specificity different from that observed for bacteria. The macroalgae L. digitata and M. stellatus exhibited more pronounced antifungal effects than C. rubrum. This study demonstrates macroalgal defenses against epiphytic microorganisms based on natural delivery mechanisms of allelochemicals utilizing a culture-independent approach, thus minimizing the ecological bias inherent to culture-dependent studies based on few microbial isolates.

Session: A-22 Free Communication/Poster - Environment, Policy, and Physical Activity (June 2, 2010)

Larval settlement of the polychaete Hydroides elegans Haswell, 1883 is mediated by marine biofilms; complex agglomerates of bacteria, diatoms, fungi and protozoa enmeshed in a matrix of extracellular polymers (EPS). In our previous investigations, benthic diatoms were demon- strated to be potent mediators of larval settlement in H. elegans. The putative diatom-derived settle- ment cues were heat-stable components in close association with the diatom surface. For an in-depth investigation of the chemical nature of diatom-derived larval settlement cues, the EPS of the induc- tive (Achnanthes sp., Nitzschia constricta) and non-inductive (Amphora tenerrima, Nitzschia frustu- lum) diatoms were bioassayed for their effect on larval settlement. When EPS fractions larger than 100 kDa were immobilized in stable hydrogels to mimic their association to a solid substratum, they evoked an effect on larval settlement similar to that of the respective monospecies diatom films. The crude exopolymer samples mainly consisted of polysaccharides with a small proportion of proteina- ceous sample components. After enzymatic removal of proteinaceous EPS components, the samples evoked an effect on larval settlement similar to that of the crude EPS samples, indicating the negligible role of large proteins or glycoproteins as settlement cues.