Brain responses to audiovisual speech mismatch in infants are associated with individual differences in looking behaviour.
Research on audiovisual speech integration has reported high levels of individual variability, especially among young infants. In the present study we tested the hypothesis that this variability results from individual differences in the maturation of audiovisual speech processing during infancy. A developmental shift in selective attention to audiovisual speech has been demonstrated between 6 and 9 months with an increase in the time spent looking to articulating mouths as compared to eyes (Lewkowicz & Hansen-Tift. (2012) Proc. Natl Acad. Sci. USA, 109, 1431-1436; Tomalski et al. (2012) Eur. J. Dev. Psychol., 1-14). In the present study we tested whether these changes in behavioural maturational level are associated with differences in brain responses to audiovisual speech across this age range. We measured high-density event-related potentials (ERPs) in response to videos of audiovisually matching and mismatched syllables /ba/ and /ga/, and subsequently examined visual scanning of the same stimuli with eye-tracking. There were no clear age-specific changes in ERPs, but the amplitude of audiovisual mismatch response (AVMMR) to the combination of visual /ba/ and auditory /ga/ was strongly negatively associated with looking time to the mouth in the same condition. These results have significant implications for our understanding of individual differences in neural signatures of audiovisual speech processing in infants, suggesting that they are not strictly related to chronological age but instead associated with the maturation of looking behaviour, and develop at individual rates in the second half of the first year of life.


Ghrelin and obestatin in human neuroendocrine tumors: expression and effect on obestatin levels after food intake.
Ghrelin and obestatin are derived from the same peptide hormone precursor and are mainly produced by the gastric mucosa. Ghrelin is involved in many biological processes, whereas the physiological function of obestatin needs further investigation. The aims of the present study were to establish the incidence of ghrelin- and obestatin-immunoreactive cells in a comprehensive panel of human neuroendocrine tumors (NETs) and to investigate if blood obestatin concentrations are influenced during a standardized meal stimulation test in healthy individuals and patients with NETs. The expression of ghrelin and obestatin was investigated in NETs (n = 149) and other endocrine-related disorders (n = 3) using immunohistochemistry with specific polyclonal antibodies. Coexpression of the peptides was evaluated by double immunofluorescence. Concentrations of obestatin in blood were measured during a meal test in 6 healthy individuals and 5 patients with pancreatic NETs. Ghrelin and obestatin were expressed in 14/152 and 19/152 tumor tissues, respectively, mainly representing NETs of foregut origin and in pancreatic tissue from a nesidioblastosis patient. Double immunofluorescence staining showed colocalization of the peptides. During the meal test, obestatin levels in blood were unchanged in all patients but decreased significantly in the healthy individuals. Only a minority of NETs express ghrelin and obestatin. However, analysis of patients with tumors originating from tissues that express the peptides in normal conditions could be of importance. The results from the meal test indicate that the hormone levels are affected by food intake in healthy individuals, whereas obestatin levels remained unchanged in pancreatic NET patients.


The effect of tibial plateau angle on ground reaction forces 4-17 months after tibial plateau leveling osteotomy in Labrador Retrievers.
To assess the relationship between postoperative tibial plateau angle (TPA) and ground reaction forces (GRFs) in Labrador Retrievers at least 4-months after tibial plateau leveling osteotomy (TPLO) surgery. A retrospective longitudinal study. Thirty-two Labrador Retrievers with unilateral cranial cruciate ligament disease that had TPLO and concurrent meniscal surgery. TPA and GRFs were measured pre- and > or = 4 months postoperatively. Preoperative GRFs, preoperative TPA, duration of injury, postoperative TPA and degree of rotation were each compared with postoperative GRFs using correlation analysis. Dogs were also grouped by postoperative TPA and compared using 1-way analysis of variance (ANOVA). Postoperative function was compared with meniscal release/meniscectomy, patient age, sex, body weight and follow-up time using ANOVA. No significant relationship was found between preoperative GRFs, preoperative TPA, duration of injury, postoperative TPA, degree of rotation, or meniscal release/meniscectomy and postoperative function. Mean preoperative GRFs for all dogs were 28.8 +/- 9.5 for peak vertical force (PVF) and 9.3 +/- 3.1 for peak vertical impulse (VI). Mean postoperative GRFs for all dogs were 40.4 +/- 5.0 for PVF and 14.1 +/- 1.9 for VI. No statistically significant relationship was found between postoperative TPA and GRFs after TPLO > or = 4 months after surgery, where the postoperative angle was between 0 and 14 degrees. No significant relationship was found between the preoperative TPA and postoperative function. Limb function in Labrador Retrievers was not affected by postoperative TPA and re-operation for cases with a postoperative TPA between 0 and 14 degrees is not recommended.


Abel inversion of knife-edge data from radially symmetric pulsed laser beams.
The intensity profile of a radially symmetric laser beam is shown to be invertible from knife-edge data via the inverse Abel transform of the first derivative of the data. Two numerical algorithms for inverting pulsed-beam data are applied to a knife-edge scan of the focused output of a confocal unstable resonator laser. The resulting beam profiles are shown to be in good agreement with each other but significantly different from the profile obtained by assuming the beam to be approximately Gaussian. The results are shown to predict that the laser damage threshold of polymethylmethacrylate is ~4 times as large as previously reported.


Experimental demonstration of an optical phased array antenna for laser space communications.
The feasibility of an optical phased array antenna applicable for spaceborne laser communications was experimentally demonstrated. Heterodyne optical phase-locked loops provide for a defined phase relationship between the collimated output beams of three single-mode fibers. In the far field the beams interfere with a measured efficiency of 99%. The main lobe of the interference pattern can be moved by phase shifting the subaperture output beams. The setup permitted agile beam steering within an angular range of 1 mr and a response time of 0.7 ms. We propose an operational optical phased array antenna fed by seven lasers, featuring high transmit power and redundance.


Novel NTRK3 Fusions in Fibrosarcomas of Adults.
NTRK fusions in malignant tumors are therapeutic targets of tyrosine kinase inhibitors. Because they occur only in a small subset of mesenchymal tumors, knowledge regarding the corresponding histology is important to effectively identify patients who could benefit from targeted therapy. In this study, using RNA sequencing, we identified novel NTRK3 fusions involving related partner genes in 2 adult bone and soft tissue tumors that met the current histologic criteria of fibrosarcoma. Case 1 involved the left radius of a 38-year-old woman, whereas in case 2, the right thigh of a 26-year-old man was affected. Histologically, both tumors consisted of the long fascicular growth of long spindle cells. The tumor in case 1 additionally showed focal myxoid changes. Tumor cells had nonpleomorphic, atypical nuclei, and lacked evidence of a specific line of differentiation. Both tumors showed widespread CD34 immunoreactivity and very limited expression of actin. RNA sequencing detected in-frame fusion transcripts of STRN (exon 3)-NTRK3 (exon 14) in case 1 and STRN3 (exon 3)-NTRK3 (exon 14) in case 2, which were confirmed by reverse transcription polymerase chain reaction and Sanger sequencing. Pan-TRK immunostaining was diffusely positive in both cases. Fluorescence in situ hybridization showed signal patterns compatible with NTRK3 rearrangements in both cases, with case 2 additionally harboring a CDKN2A homozygous deletion. This study expands the clinicopathologic and genetic spectrum of sarcomas associated with NTRK fusions, and suggests that CD34-positive fibrosarcoma of bone and soft tissue could be a good candidate for NTRK testing.


Ribonuclease A suggests how proteins self-chaperone against amyloid fiber formation.
Genomic analyses have identified segments with high fiber-forming propensity in many proteins not known to form amyloid. Proteins are often protected from entering the amyloid state by molecular chaperones that permit them to fold in isolation from identical molecules; but, how do proteins self-chaperone their folding in the absence of chaperones? Here, we explore this question with the stable protein ribonuclease A (RNase A). We previously identified fiber-forming segments of amyloid-related proteins and demonstrated that insertion of these segments into the C-terminal hinge loop of nonfiber-forming RNase A can convert RNase A into the amyloid state through three-dimensional domain-swapping, where the inserted fiber-forming segments interact to create a steric zipper spine. In this study, we convert RNase A into amyloid-like fibers by increasing the loop length and hence conformational freedom of an endogenous fiber-forming segment, SSTSAASS, in the N-terminal hinge loop. This is accomplished by sandwiching SSTSAASS between inserted Gly residues. With these inserts, SSTSAASS is now able to form the steric zipper spine, allowing RNase A to form amyloid-like fibers. We show that these fibers contain RNase A molecules retaining their enzymatic activity and therefore native-like structure. Thus, RNase A appears to prevent fiber formation by limiting the conformational freedom of this fiber-forming segment from entering a steric zipper. Our observations suggest that proteins have evolved to self-chaperone by using similar protective mechanisms.


Wheat heat tolerance is impaired by heightened deletions in the distal end of 4AL chromosomal arm.
Heat stress (HS) causes substantial damages to worldwide crop production. As a cool season crop, wheat (Triticum aestivum) is sensitive to HS-induced damages. To support the genetic improvement of wheat HS tolerance (HST), we conducted fine mapping of TaHST1, a locus required for maintaining wheat vegetative and reproductive growth under elevated temperatures. TaHST1 was mapped to the distal terminus of 4AL chromosome arm using genetic populations derived from two BC6 F6 breeding lines showing tolerance (E6015-4T) or sensitivity (E6015-3S) to HS. The 4AL region carrying TaHST1 locus was approximately 0.949 Mbp and contained the last 19 high confidence genes of 4AL according to wheat reference genome sequence. Resequencing of E6015-3S and E6015-4T and haplotype analysis of 3087 worldwide wheat accessions revealed heightened deletion polymorphisms in the distal 0.949 Mbp region of 4AL, which was confirmed by the finding of frequent gene losses in this region in eight genome-sequenced hexaploid wheat cultivars. The great majority (86.36%) of the 3087 lines displayed different degrees of nucleotide sequence deletions, with only 13.64% of them resembling E6015-4T in this region. These deletions can impair the presence and/or function of TaHST1 and surrounding genes, thus rendering global wheat germplasm vulnerable to HS or other environmental adversities. Therefore, conscientious and urgent efforts are needed in global wheat breeding programmes to optimize the structure and function of 4AL distal terminus by ensuring the presence of TaHST1 and surrounding genes. The new information reported here will help to accelerate the ongoing global efforts in improving wheat HST.


Robotic radical cystectomy with intracorporeal urinary diversion: beyond the initial experience.
Robotic assisted radical cystectomy (RARC) has gained popularity within minimally-invasive urologic surgery, and has been shown to be a safe procedure with similar oncologic outcomes when compared to the conventional open standard. While initial RARC feasibility and outcomes studies were performed with extracorporeal urinary diversion, intracorporeal urinary diversion (ICUD) is becoming increasingly utilized. Reported benefits of an intracorporeal approach include decreased blood loss and a lower incidence of ureteral strictures. While ICUD is technically challenging, many have overcome the learning curve associated with this procedure via a mentorship model and a dedicated operative team. Techniques vary between institutions, and ileal conduit, continent cutaneous and orthotopic continent (neobladder) diversions have all been performed. Herein, we describe the learning curve, technical points, and unique complications associated with ICUD.


Lymphoepithelial cysts of the pancreas: the use of endoscopic ultrasound-guided fine-needle aspiration in diagnosis.
Lymphoepithelial cysts (LECs) are rare non-neoplastic lesions that can appear as a complex cyst or a mass in the pancreas. Cytology from endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) can be helpful in making a diagnosis with the aim of avoiding unnecessary surgical resection. A case involving a 51-year-old woman with lower abdominal pain who was found to have a multiloculated cystic lesion at the junction of the pancreatic body and tail is described. Cytology from EUS-FNA was consistent with a pancreatic LEC. The lesion was managed conservatively and follow-up imaging of the cyst over the following two years was unchanged. The patient remains clinically well. Cytology from EUS-FNA can help distinguish LECs from cystic neoplasms, thus preventing radical surgical resection of this benign pancreatic cyst.


Kinetic properties of alpha 1 beta 1 gamma-aminobutyric acidA receptor channels expressed in Chinese hamster ovary cells: regulation by pentobarbital and picrotoxin.
Single-channel recordings from excised outside-out patches were obtained from Chinese hamster ovary cells stably transfected with plasmids containing bovine gamma-aminobutyric acid (GABA) type A (GABAA) receptor channel alpha 1 and beta 1 subunit cDNAs. The predominant or main conductance level recorded had a 17-pS chord conductance. There were minor contributions made from 25-pS and 11-pS conductance levels. Average open duration, burst duration, and openings/burst did not change as the GABA concentration was increased from 5 to 25 microM. However, opening frequency increased from 11.0 to 19.5 openings/sec. Pentobarbital increased average channel open duration without increasing opening frequency, whereas picrotoxin slightly reduced average channel open duration and reduced opening frequency. Open duration frequency distributions were fitted best with the sum of two exponential functions, suggesting that the alpha 1 beta 1 GABAA receptor channel had at least two open states. The time constants and relative proportions of the two components did not vary when GABA concentration was increased from 5 to 25 microM. Closed duration distributions of closures between main conductance level openings were fitted best with multiple exponential functions, suggesting that the alpha 1 beta 1 GABAA receptor channel had several closed states. Burst duration frequency distributions were fitted best with two exponential functions whose time constants and relative proportions did not change with GABA concentration. A gating kinetic scheme for the alpha 1 beta 1 GABAA receptor channel was proposed that consisted of a single binding site for GABA and at least two open and five closed states. The kinetic properties of the alpha 1 beta 1 main conductance level differed from those of the spinal cord neuron (native) 27-pS main conductance level and the 19-pS subconductance level. The native main conductance and subconductance levels were characterized by longer openings and at least three open states. Based on the aforementioned observations, it appears that different subunit combinations produce receptor channels with different kinetic properties, but the basic mechanism of regulation by pentobarbital and picrotoxin may be similar for the different receptor channels. Also, it is unlikely that the 19-pS substate of the native GABAA receptor is produced by an alpha 1 beta 1 dimer.


Spatial Effects on the Multiplicity of Plasmodium falciparum Infections.
As malaria is being pushed back on many frontiers and global case numbers are declining, accurate measurement and prediction of transmission becomes increasingly difficult. Low transmission settings are characterised by high levels of spatial heterogeneity, which stands in stark contrast to the widely used assumption of spatially homogeneous transmission used in mathematical transmission models for malaria. In the present study an individual-based mathematical malaria transmission model that incorporates multiple parasite clones, variable human exposure and duration of infection, limited mosquito flight distance and most importantly geographically heterogeneous human and mosquito population densities was used to illustrate the differences between homogeneous and heterogeneous transmission assumptions when aiming to predict surrogate indicators of transmission intensity such as population parasite prevalence or multiplicity of infection (MOI). In traditionally highly malaria endemic regions where most of the population harbours malaria parasites, humans are often infected with multiple parasite clones. However, studies have shown also in areas with low overall parasite prevalence, infection with multiple parasite clones is a common occurrence. Mathematical models assuming homogeneous transmission between humans and mosquitoes cannot explain these observations. Heterogeneity of transmission can arise from many factors including acquired immunity, body size and occupational exposure. In this study, we show that spatial heterogeneity has a profound effect on predictions of MOI and parasite prevalence. We illustrate, that models assuming homogeneous transmission underestimate average MOI in low transmission settings when compared to field data and that spatially heterogeneous models predict stable transmission at much lower overall parasite prevalence. Therefore it is very important that models used to guide malaria surveillance and control strategies in low transmission and elimination settings take into account the spatial features of the specific target area, including human and mosquito vector distribution.


K+ pore structure revealed by reporter cysteines at inner and outer surfaces.
The structure of the carboxyl half of the pore-forming region of Kv2.1 was studied by replacing each of 15 consecutive residues between positions 383 and 369 with a reporter cysteine residue. Extracellular application of charged, membrane-impermeant methanethiosulfonates irreversibly modified currents at four cysteine-substituted positions, K382, Y380, I379, and D378. Intracellular exposure to methanethiosulfonate ethyltrimethylammonium revealed another set of reactive mutants (V374, T373, T372, and T370). Our results indicate that positions 378 and 374 are exposed at outer and inner mouths of the channel, respectively, and immersed in the aqueous phase. In contrast to present topological models, the 383-369 region appears to span the pore mainly as a nonperiodic structure.


Intrinsic single-domain switching in ferroelectric materials on a nearly ideal surface.
Ferroelectric domain nucleation and growth in multiferroic BiFeO(3) is studied on a single-domain level by using piezoresponse force spectroscopy. Variation of local electromechanical response with dc tip bias is used to determine the size of the domain formed below the conductive scanning probe tip. The domain parameters are calculated self-consistently from the decoupled Green function theory by using tip geometry determined from the domain wall profile. The critical parameters of the nucleating domain and the activation energy for nucleation are determined. The switching mechanism is modeled by using the phase-field method, and comparison with experimental results shows that the nucleation biases are within a factor of approximately 2 of the intrinsic thermodynamic limit. The role of atomic-scale defects and long-range elastic fields on nucleation bias lowering is discussed. These measurements open a pathway for quantitative studies of the role of a single defect on kinetics and thermodynamics of first order bias-induced phase transitions and electrochemical reactions.


Curriculum for pharmacology in pharmacy institutions in India: opportunities and challenges.
The curriculum of pharmacy institutions in India is regulated by the All India Council for Technical Education (AICTE) and the Pharmacy Council of India (PCI) at degree and diploma levels. However, it has been over two decades that the syllabi have been revised by these regulatory agencies. Considering the dynamic character of pharmacology, it is essential to prepare a syllabus that caters to the contemporary needs of the academic institutions and pharmaceutical industry, the community. Pharmacists are also witnessing a greater role in community pharmacy practice as well as in several healthcare sectors. Considering these facts, a panel discussion was held at IPSCON 2013, (the Annual Conference of Indian Pharmacological Society) at Bangalore. The discussion saw several recommendations for syllabi for institutions offering various pharmacy courses to meet the objectives of teaching, learning and research in Pharmacology. This article documents a summary of the discussion. For B. Pharm. course, a balance between industry-oriented pharmacology and clinical pharmacy has been recommended. Redundant animal experiments should be replaced with the simulation experiments or those which are feasible in the light of stringent regulations of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA). It is recommended that the M. Pharm curriculum should focus on preclinical research with the inclusion of molecular biology and experiments on gene expression, proteomics, pharmacogenomics, cell culture and tissue culture. In general, at all levels, exposure of students to hospitals and clinicians is needed. Pharm. D., syllabus too should lay lesser emphasis on experimental pharmacology. Present experiments in the D. Pharm. course have no relevance to the program objectives and hence, only experiments through demonstrations or simulated preparations or interactive videos maybe undertaken. Regulatory bodies as well as universities should design a comprehensive syllabus and plan an effective pedagogy to prepare graduates who are competent and capable of bringing positive changes in the community and healthcare in India.


Development and evaluation of novel PET tracers for imaging cannabinoid receptor type 2 in brain.
The cannabinoid receptor type 2 (CB2) has a very low expression level in brain tissue under basal conditions, but it is up-regulated in diverse pathological conditions. Two promising lead structures from the literature, N-((3S,5S,7S)-adamantan-1-yl)-8-methoxy-4-oxo-1-pentyl-1,4-dihydroquinoline-3-carboxamide and 8-butoxy-N-(2-fluoro-2-phenylethyl)-7-methoxy-2-oxo-1,2-dihydroquinoline-3-carboxamide - designated KD2 and KP23, respectively - were evaluated as potential PET ligands for imaging CB2. Both KD2 and KP23 were synthesized and labeled with carbon-11. In vitro autoradiographic studies on rodent spleen tissues showed that [(11)C]KD2 exhibits superior properties. A pilot study using [(11)C]KD2 on human post mortem ALS spinal cord slices indicated high CB2 expression level and specific binding, a very exciting finding if considering the future diagnostic application of CB2 ligands and their utility in therapy monitoring. In vivo blocking studies in rats with [(11)C]KD2 showed also high specific uptake in spleen tissue. Although the protein-bound fraction is relatively high, KD2 or KD2 derivatives could be very useful tools for the non-invasive investigation of CB2 levels under various neuroinflammatory conditions.


Focus issue: the evolution of complexity.
With genomic and proteomic data sets in hand, scientists are beginning to understand how variation in signaling proteins and signaling networks influences the function of signaling pathways to regulate cellular behavior. These analyses have provided insight into how proteins have evolved, how biological networks have evolved, and how organisms have evolved. Not only do these discoveries inform us about evolution of life at different levels, but they also have the potential to help us understand human disease and begin to engineer signaling networks to achieve specified tasks.


Insights into How Longicorn Beetle Larvae Determine the Timing of Metamorphosis: Starvation-Induced Mechanism Revisited.
Larvae of holometabolous insects must determine the timing of their metamorphosis. How they determine this timing has only been studied in detail for a few insect species. In a few species of Coleoptera, starvation is known to be a cue for metamorphosis, leading to the formation of smaller adults (starvation-induced pupation, SiP). We investigated the occurrence of SiP in the beetle Psacothea hilaris. When P. hilaris larvae were starved late in the feeding phase of the last (5th) instar, they exhibited typical SiP characterized by constancy of the time from food deprivation to pupation (TTP) irrespective of the body weight upon food deprivation or the length of prior feeding. In contrast, when larvae were starved early in the feeding phase, TTP decreased by roughly 1 day as the feeding became 1 day longer. The change in the response to starvation was estimated to occur on day 5.9 in the last instar. A series of refeeding experiments suggested that whereas SiP occurred readily in the larvae starved in the late feeding phase, activation of SiP was suspended until day 5.9 in the larvae starved early in the feeding phase. When P. hilaris larvae were fed continuously, they eventually ceased feeding spontaneously and pupated. The time length between spontaneous cessation of feeding and pupation was approximately equal to the TTP in SiP. This suggests that the same mechanism was activated by food deprivation in the late feeding phase and by spontaneous cessation of ad libitum feeding.


A simian-adenovirus-vectored rabies vaccine suitable for thermostabilisation and clinical development for low-cost single-dose pre-exposure prophylaxis.
Estimates of current global rabies mortality range from 26,000 to 59,000 deaths per annum. Although pre-exposure prophylaxis using inactivated rabies virus vaccines (IRVs) is effective, it requires two to three doses and is regarded as being too expensive and impractical for inclusion in routine childhood immunization programmes. Here we report the development of a simian-adenovirus-vectored rabies vaccine intended to enable cost-effective population-wide pre-exposure prophylaxis against rabies. ChAdOx2 RabG uses the chimpanzee adenovirus serotype 68 (AdC68) backbone previously shown to achieve pre-exposure protection against rabies in non-human primates. ChAdOx2 differs from AdC68 in that it contains the human adenovirus serotype 5 (AdHu5) E4 orf6/7 region in place of the AdC68 equivalents, enhancing ease of manufacturing in cell lines which provide AdHu5 E1 proteins in trans. We show that immunogenicity of ChAdOx2 RabG in mice is comparable to that of AdC68 RabG and other adenovirus serotypes expressing rabies virus glycoprotein. High titers of rabies virus neutralizing antibody (VNA) are elicited after a single dose. The relationship between levels of VNA activity and rabies virus glycoprotein monomer-binding antibody differs after immunization with adenovirus-vectored vaccines and IRV vaccines, suggesting routes to further enhancement of the efficacy of the adenovirus-vectored candidates. We also demonstrate that ChAdOx2 RabG can be thermostabilised using a low-cost method suitable for clinical bio-manufacture and ambient-temperature distribution in tropical climates. Finally, we show that a dose-sparing effect can be achieved by formulating ChAdOx2 RabG with a simple chemical adjuvant. This approach could lower the cost of ChAdOx2 RabG and other adenovirus-vectored vaccines. ChAdOx2 RabG may prove to be a useful tool to reduce the human rabies death toll. We have secured funding for Good Manufacturing Practice- compliant bio-manufacture and Phase I clinical trial of this candidate.


Acute mental health care and South African mental health legislation: part 1- morbidity, treatment and outcome.
This is the first of three reports on a follow-up review of mental health care at Helen Joseph Hospital (HJH). In this first part, qualitative and quantitative descriptions were made of the services and of demographic and clinical data on acute mental health care users managed at HJH, in a retrospective review of clinical records over a four year period. Objectives for this review were to provide information on mental health care outcome, to do a cost analysis and to establish a quality assurance cycle that may facilitate a cost centre management approach. The operational areas identified were service delivery, teaching, and research. Activities within each area were in-patient care, out-patients and consultation/liaison, under- and postgraduate teaching and self initiated or contract research. The study reviewed the existing mental health care program and activities in context of relevant policy and legislation. Norms from a World Health Organization model for acute mental health care showed that significant staff shortages existed, especially for nursing. A total of 520 users were admitted for in-patient mental health care during the financial year 2007/08. The average length of stay was 15.4 days and ranged from 1 to 85 days. Ninety users (17%) had an extended period of stay of 25 days and more, while 39 users had multiple admissions during the 12 month period. The most common Axis I diagnoses made were schizophrenia n=138 (29%), substance-related conditions n=99 (21%) and bipolar mood disorder n=69 (14%). After discharge, 139 users (27%) were referred back to the HJH out-patient department for follow-up. The information from these reports may be used in the allocation of adequate resources to align this acute unit with its responsibilities according to recent legislation.


Socially sensitive lactation: exploring the social context of breastfeeding.
Many women report difficulties with breastfeeding and do not maintain the practice for as long as intended. Although psychologists and other researchers have explored some of the difficulties they experience, fuller exploration of the relational contexts in which breastfeeding takes place is warranted to enable more in-depth analysis of the challenges these pose for breastfeeding women. This article is based on qualitative data collected from 22 first-time breastfeeding mothers through two phases of interviews and audio-diaries which explored how the participants experienced their relationships with significant others and the wider social context of breastfeeding in the first five weeks postpartum. Using a thematic analysis informed by symbolic interactionism, we develop the overarching theme of 'Practising socially sensitive lactation' which captures how participants felt the need to manage tensions between breastfeeding and their perceptions of the needs, expectations and comfort of others. We argue that breastfeeding remains a problematic social act, despite its agreed importance for child health. While acknowledging the limitations of our sample and analytic approach, we suggest ways in which perinatal and public health interventions can take more effective account of the social challenges of breastfeeding in order to facilitate the health and psychological well-being of mothers and their infants.


Protection against β-amyloid-induced synaptic and memory impairments via altering β-amyloid assembly by bis(heptyl)-cognitin.
β-amyloid (Aβ) oligomers have been closely implicated in the pathogenesis of Alzheimer's disease (AD). We found, for the first time, that bis(heptyl)-cognitin, a novel dimeric acetylcholinesterase (AChE) inhibitor derived from tacrine, prevented Aβ oligomers-induced inhibition of long-term potentiation (LTP) at concentrations that did not interfere with normal LTP. Bis(heptyl)-cognitin also prevented Aβ oligomers-induced synaptotoxicity in primary hippocampal neurons. In contrast, tacrine and donepezil, typical AChE inhibitors, could not prevent synaptic impairments in these models, indicating that the modification of Aβ oligomers toxicity by bis(heptyl)-cognitin might be attributed to a mechanism other than AChE inhibition. Studies by using dot blotting, immunoblotting, circular dichroism spectroscopy, and transmission electron microscopy have shown that bis(heptyl)-cognitin altered Aβ assembly via directly inhibiting Aβ oligomers formation and reducing the amount of preformed Aβ oligomers. Molecular docking analysis further suggested that bis(heptyl)-cognitin presumably interacted with the hydrophobic pockets of Aβ, which confers stabilizing powers and assembly alteration effects on Aβ. Most importantly, bis(heptyl)-cognitin significantly reduced cognitive impairments induced by intra-hippocampal infusion of Aβ oligomers in mice. These results clearly demonstrated how dimeric agents prevent Aβ oligomers-induced synaptic and memory impairments, and offered a strong support for the beneficial therapeutic effects of bis(heptyl)-cognitin in the treatment of AD.


Inhibitor of Apoptosis Protein-1 Regulates Tumor Necrosis Factor-Mediated Destruction of Intestinal Epithelial Cells.
Tumor necrosis factor (TNF) is a cytokine that promotes inflammation and contributes to pathogenesis of inflammatory bowel diseases. Unlike other cells and tissues, intestinal epithelial cells undergo rapid cell death upon exposure to TNF, by unclear mechanisms. We investigated the roles of inhibitor of apoptosis proteins (IAPs) in the regulation of TNF-induced cell death in the intestinal epithelium of mice and intestinal organoids. RNA from cell lines and tissues was analyzed by quantitative polymerase chain reaction, protein levels were analyzed by immunoblot assays. BIRC2 (also called cIAP1) was expressed upon induction from lentiviral vectors in young adult mouse colon (YAMC) cells. YAMC cells, the mouse colon carcinoma cell line MC38, the mouse macrophage cell line RAW 264.7, or mouse and human organoids were incubated with second mitochondrial activator of caspases (Smac)-mimetic compound LCL161 or recombinant TNF-like weak inducer of apoptosis (TNFSF12) along with TNF, and cell death was quantified. C57BL/6 mice with disruption of Xiap, Birc2 (encodes cIAP1), Birc3 (encodes cIAP2), Tnfrsf1a, or Tnfrsf1b (Tnfrsf1a and b encode TNF receptors) were injected with TNF or saline (control); liver and intestinal tissues were collected and analyzed for apoptosis induction by cleaved caspase 3 immunohistochemistry. We also measured levels of TNF and alanine aminotransferase in serum from mice. YAMC cells, and mouse and human intestinal organoids, died rapidly in response to TNF. YAMC and intestinal crypts expressed lower levels of XIAP, cIAP1, cIAP2, and cFLIP than liver tissue. Smac-mimetics reduced levels of cIAP1 and XIAP in MC38 and YAMC cells, and Smac-mimetics and TNF-related weak inducer of apoptosis increased TNF-induced cell death in YAMC cells and organoids-most likely by sequestering and degrading cIAP1. Injection of TNF greatly increased levels of cell death in intestinal tissue of cIAP1-null mice, compared with wild-type C57BL/6 mice, cIAP2-null mice, or XIAP-null mice. Excessive TNF-induced cell death in the intestinal epithelium was mediated TNF receptor 1. In a study of mouse and human cell lines, organoids, and tissues, we found cIAP1 to be required for regulation of TNF-induced intestinal epithelial cell death and survival. These findings have important implications for the pathogenesis of TNF-mediated enteropathies and chronic inflammatory diseases of the intestine.


Nivolumab for relapsed or refractory Hodgkin lymphoma: real-life experience.
Classical Hodgkin lymphoma (cHL) is considered a curable disease; however, in approximately one-third of the responding patients, the disease relapses following completion of therapy. One of the drugs that have been approved for the treatment of relapsed/refractory cHL is nivolumab, an immune check point inhibitor that shows its effects by blocking the programmed death 1 (PD-1) receptor. In this study, we present a retrospective "real-life" analysis of the usage of nivolumab in patients with relapsed/refractory cHL that have joined the named patient program (NPP) for nivolumab, reflecting 4 years of experience in the treatment of relapsed/refractory cHL. We present a retrospective analysis of 87 patients (median age, 30) that participated in the NPP in 24 different centers, who had relapsed/refractory cHL and were consequently treated with nivolumab. The median follow-up was 29 months, and the median number of previous treatments was 5 (2-11). In this study, the best overall response rate was 70% (CR, 36%; PR, 34%). Twenty-eight of the responding patients underwent subsequent stem cell transplantation (SCT). Among 15 patients receiving allogeneic stem cell transplantation, 9 patients underwent transplantation with objective response, of which 8 of them are currently alive with ongoing response. At the time of analysis, 23 patients remained on nivolumab treatment and the rest discontinued therapy. The main reason for discontinuing nivolumab was disease progression (n = 23). The safety profile was acceptable, with only nine patients requiring cessation of nivolumab due to serious adverse events. The 24-month progression-free and overall survival rates were 58.5% (95% CI, 0.47-0.68) and 78.7% (95% CI, 0.68-0.86), respectively. Eighteen patients died during the follow-up and only one of these was regarded to be treatment-related. With its efficacy and its safety profile, PD-1 blockers became an important treatment option in the heavily pretreated cHL patients.


TIPS for the treatment of refractory ascites, hepatorenal syndrome and hepatic hydrothorax: a critical update.
Refractory ascites is a frequent complication of advanced cirrhosis and is associated with hepatorenal syndrome and hepatic hydrothorax. Large volume paracentesis and pleurodesis are regarded as first-line treatments in patients who do not respond adequately to diuretics. These treatments, however, do not prevent recurrence and carry the risk of worsening of the circulatory dysfunction leading to hepatorenal syndrome. The transjugular intrahepatic portosystemic shunt (TIPS) has been proposed as an alternative to paracentesis. TIPS reduces the rate of ascites recurrence mainly due to the reduction in the filtration pressure. In addition, TIPS results in a positive effect on renal function, including hepatorenal syndrome, demonstrated by a rapid increase in urinary sodium excretion, urinary volume, and improvement in plasma creatinine concentration. Furthermore, plasma renin activity, aldosterone, and noradrenalin concentrations improve gradually after TIPS insertion suggesting a positive effect on systemic underfilling, the factor of hepatorenal syndrome. As demonstrated recently in two meta-analyses including five randomised studies, TIPS also improves survival when compared with paracentesis. However, the evidence is based on relatively few studies with only 305 patients included. The positive effects of the TIPS are opposed by an increased frequency and severity of episodes of hepatic encephalopathy which may be reduced by both patient selection and reduced shunt diameter. Based on the present knowledge the recommended hierarchy of treatments for refractory ascites may be reconsidered upgrading TIPS in suitable candidates.


Green synthesis of lead sulfide nanoparticles by the lead resistant marine yeast, Rhodosporidium diobovatum.
Biosynthesis of nanoparticles using microorganisms has attracted a lot of attention in recent years as this route has the potential to lead to synthesis of monodisperse nanoparticles. Here, we report the intracellular synthesis of stable lead sulfide nanoparticles by a marine yeast, Rhodosporidium diobovatum. The PbS nanoparticles were characterized by UV-visible absorption spectroscopy, X-ray diffraction (XRD) and energy dispersive atomic spectroscopy (EDAX). UV-visible absorption scan revealed a peak at 320 nm, a characteristic of the nanosize range. XRD confirmed the presence of PbS nanoparticles of cubic structure. Crystallite size as determined from transmission electron microscopy was found to be in the range of 2-5 nm. Elemental analysis by EDAX revealed the presence of particles composed of lead and sulfur in a 1:2 ratio indicating that PbS nanoparticles were capped by a sulfur-rich peptide. A quantitative study of lead uptake through atomic absorption spectrometry revealed that 55% of lead in the medium was accumulated in the exponential phase, whereas a further 35% was accumulated in the stationary phase; thus, the overall recovery of PbS nanoparticles was 90%. The lead-exposed yeast displayed a marked increase (280% over the control) in nonprotein thiols in the stationary phase.


Early cell death in the brain of fetal preterm lambs after hypoxic-ischemic injury.
The objective of the present study was to evaluate using premature fetal lambs the effect of cerebral hypoxia-ischemia induced by partial occlusion of the umbilical cord on the type of cell death which occurs in different brain regions and to ascertain some of the neural pathways which may underlie the associated pathologies. Lambs were sacrificed either immediately after a 1 h hypoxic-ischemic insult or 3 h later. Brains were fixed by perfusion and blocks of the different brain territories were processed for light microscopy (hematoxylin-eosin, Nissl staining), electron transmission microscopy and quantification of apoptosis by the TUNEL method. Other fixed brains were dissociated and labeled by nonyl acridine orange to determine mitochondrial integrity. Non-fixed brains were also used for membrane asymmetry studies, in which cell suspensions were analyzed by flow cytometry to quantify apoptosis. In both hypoxic-ischemic groups, necrotic-like neurons were observed mainly in the mesencephalon, pons, deep cerebellar nuclei and basal nuclei, whereas apoptotic cells were extensively found both in white and gray matter and were not limited to regions where necrotic neurons were present. The 3 h post-partial cord occlusion group, but not the 0 h group, showed a generalized alteration of cell membrane asymmetry and mitochondrial integrity as revealed by Annexin V/PI flow cytometry and nonyl acridine orange studies, respectively. Our results show that the apoptotic/necrotic patterns of cell death occurring early after hypoxic-ischemic injury are brain-region-specific and have distinct dynamics and suggest that therapeutic strategies aimed at rescuing cells from the effects of hypoxia/ischemia should be aimed at blocking the apoptotic components of brain damage.


Variants of human chorionic gonadotropin from pregnant women and tumor patients recognized by monoclonal antibodies.
In biological fluids, hCG and its free alpha- (hCG alpha) and beta-subunits (hCG beta), occur in multiple forms. These various forms differ at the molecular level primarily in glycosylation, but also differ in protein backbone modifications corresponding to the urinary low molecular weight fragment of the hCG beta-subunit (beta-core fragment). This microheterogeneous nature can be demonstrated by isoelectric focusing in which variants are separated into bands with different isoelectric points (pI). To determine whether such isoelectric variants differ in antigenicity and consequently might escape immunoassay detection due to overspecificity of monoclonal antibodies (MCA), urinary pregnancy hCG (NIH, CR123) and tumor hCG preparations, such as a tumor-specific acidic variant of hCG (hCGav) and the hCG beta-core fragment, were separated by isoelectric focusing in the absence or presence of 8 M urea, or by sodium docedyl sulfate-polyacrylamide gel electrophoresis and enzymatically immunostained using an MCA panel directed against 17 different hCG epitopes. MCA against 14 different epitopes accessible on holo-hCG recognized all pI variants of pregnancy holo-hCG or tumor-derived hCGav, as was true for the three MCA recognizing epitopes hidden on holo-hCG but accessible on the free subunits after hCG dissociation by urea. We conclude that each individual pI-isoform of holo-hCG and its free subunits expresses the entire set of epitopes recognized by our MCA panel. The carbohydrate moieties that form a biochemical basis for hCG heterogeneity seem to be neither of major antigenic relevance, nor are they structurally related to any particular epitope. Thus, various glycosylation forms of hCG, hCG alpha, hCG beta, and hCG beta-core in normal as well as in pathological samples should safely be detectable and measureable by immunoassays employing MCA with appropriate subunit specificity.


Asymmetric synthesis of (-)-naltrexone.
(-)-Naltrexone, an opioid antagonist used extensively for the management of drug abuse, is derived from naturally occurring opioids. Herein, we report the first asymmetric synthesis of (-)-naltrexone that does not proceed through thebaine. The synthesis starts with simple, achiral precursors with catalytic enantioselective Sharpless dihydroxylation employed to introduce the stereogenic centers. A Rh(i)-catalyzed C-H alkenylation and torquoselective electrocyclization cascade provides the hexahydro isoquinoline bicyclic framework that serves as the precursor to the morphinan core. The acidic conditions used for Grewe cyclization not only provide the morphinan framework, but also cause a hydride shift resulting in the introduction of the C-6 oxo functionality present in (-)-naltrexone. The C-14 hydroxyl group is installed by an efficient two-step sequence of Pd-mediated ketone to enone dehydrogenation followed by C-H allylic oxidation using Cu(ii) and O2, a method that has not previously been reported either for the synthesis or semi-synthesis of opioids. The longest linear sequence is 17 steps, and because the stereogenic centers in the product rely on Sharpless asymmetric dihydroxylation, the route could be used to access either enantiomer of the natural product, which have disparate biological activities. The route also may be applicable to the preparation of opioid derivatives that could not be easily prepared from the more fully elaborated and densely functionalized opioid natural products that have traditionally served as the starting inputs.


SRC-3 transcription-coupled activation, degradation, and the ubiquitin clock: is there enough coactivator to go around in cells?
Overexpression of nuclear receptor coactivators is a frequent event in breast cancer cells and is recognized as a key mechanism for these cells to maximize their oncogenic growth state. Steroid receptor coactivator-3 [(SRC-3), also known as amplified in breast cancer-1 or AIB1] is foremost among these overexpressed oncogenic coactivators, being overexpressed in most breast cancers. Because of its oncogenic potential, normal cells must carefully control its cellular concentration. We discuss how SRC-3 quantitatively influences estrogen-regulated gene transcription when it is at limiting concentrations in normal breast cells and at nonlimiting concentrations in breast cancer cells. Precise control of the cellular concentration of SRC-3 may thus serve as a mechanism for defining growth responses to estrogen receptors and other growth-promoting transcription factors.


Hyperglycemia predicts unfavorable outcomes in acute ischemic stroke patients treated with intravenous thrombolysis among a Chinese population: A prospective cohort study.
Poorly controlled blood glucose was reported to cause deterioration of acute ischemic stroke. In this study, we investigated whether an elevated admission serum glucose level in the 3-h time window of intravenous thrombolysis for acute ischemic stroke determined poor functional outcomes among Chinese population. This was a prospective cohort study. From December 1, 2004 to December 31, 2016, a total of 2370 patients were enrolled and categorized into two cohorts by blood glucose levels of ≥200 and <200 mg/dl in the 3 h after stroke onset. The primary objective was to determine whether admission hyperglycemia increased major disability and death at 30 and 90 days, which was defined by a modified Rankin Scale of 3-6. The secondary objective was to determine whether admission hyperglycemia increased the symptomatic intracranial hemorrhage (SICH) at 90 days. The number needed to harm (NNH) and patient expected event rate (PEER) were evaluated for both the primary and secondary objectives. The primary outcome occurred in 216 of 305 patients (70.8%) in the blood glucose ≥200 mg/dl cohort and in 951 of 1587 patients (59.9%) in the blood glucose <200 mg/dl cohort at 30 days, and in 191 of 287 patients (66.6%) in the blood glucose ≥200 mg/dl cohort and in 827 of 1517 patients (54.5%) in the blood glucose <200 mg/dl cohort at 90 days. Patients with admission hyperglycemia had significantly increased major disability and death at 30 (adjusted relative risk (RR): 1.194 [1.073-1.329], p = 0.0012) and 90 days (adjusted RR: 1.203 [1.079-1.340], p = 0.0008). Admission hyperglycemia increased the risk of the occurrence of SICH by nearly 2-fold (adjusted RR: 1.891 [0.977-3.657], p = 0.0585 with the SITS-MOST criteria and adjusted RR: 1.884 [1.138-3.121], p = 0.0139 with the NINDS criteria). NNH values of admission hyperglycemia in major disability and death at 30 and 90 days were 9 and 10, and NNH values of SICH by the SITS-MOST NINDS criteria were 44 and 34, respectively. The study evidenced the association and temporal relationship of admission hyperglycemia causing deterioration of functional outcomes and increased SICH among Chinese population with acute ischemic stroke treated with intravenous thrombolysis.


Safety of insulin tolerance test for the assessment of growth hormone deficiency in children.
To determine the safety of insulin tolerance test (ITT) for assessing growth hormone (GH) deficiency in children. This hospital based study was conducted at the National Institute of Child Health, Karachi from 1st November 2008 till 30th October 2009. All children suspected of growth hormone deficiency, were included after excluding all other causes of short stature. Verbal informed consent was taken from the parents. Children less than 2 years of age, weighing less than 10 kg, untreated/inadequately treated hypothyroidism or Addison's disease, epilepsy, having history of hypoglycaemic fits or cardiac disease were excluded. All children were subjected to the international standard protocol of ITT and their samples of growth hormone and blood sugars were drawn. Complications during the procedure like hypoglycaemia, hypothermia, loss of consciousness, fits, vomiting and failure to achieve hypoglycaemia were recorded. Insulin tolerance test was performed on a total of 168 subjects. The data was entered in SPSS version 17 for analysis. A total of 168 children were subjected to the ITT. Four of them were abandoned as they could not achieve hypoglycaemia despite repeating the dose of insulin. Results were analyzed on 164 children whose mean age was 10 +/- 3.5 years, There were 96 (58%) males and 68 (41%) females. Over all 79.8% children achieved hypoglycaemia. None of the subjects developed any complication (fits, loss of consciousness,) or required intravenous glucose during the test and it was completed in all children with close monitoring. The results showed that there was a significant effect of time after insulin administration on both the blood glucose level (BG) and growth hormone (GH) levels. The blood glucose level decreased rapidly after administration of insulin and was lowest 30 minutes after injection and showed an increasing trend in subsequent readings, becoming almost equal to the baseline value 120 min after injection. From the study group 111 (66%) children were diagnosed as having growth hormone deficiency, 52 (31.3%) were normal and 1 (0.6%) had growth hormone insensitivity. ITT in children was found to be a safe and reliable test but can be potentially dangerous and requires very close monitoring and supervision and should be performed in a center with experienced staff.


Tempo and mode of genome evolution in a 50,000-generation experiment.
Adaptation by natural selection depends on the rates, effects and interactions of many mutations, making it difficult to determine what proportion of mutations in an evolving lineage are beneficial. Here we analysed 264 complete genomes from 12 Escherichia coli populations to characterize their dynamics over 50,000 generations. The populations that retained the ancestral mutation rate support a model in which most fixed mutations are beneficial, the fraction of beneficial mutations declines as fitness rises, and neutral mutations accumulate at a constant rate. We also compared these populations to mutation-accumulation lines evolved under a bottlenecking regime that minimizes selection. Nonsynonymous mutations, intergenic mutations, insertions and deletions are overrepresented in the long-term populations, further supporting the inference that most mutations that reached high frequency were favoured by selection. These results illuminate the shifting balance of forces that govern genome evolution in populations adapting to a new environment.


Why do physicians prescribe stress ulcer prophylaxis to general medicine patients?
Little is known about why physicians prescribe inappropriate stress ulcer prophylaxis (SUP) among nonintensive care unit (ICU) hospitalized patients without supporting evidence. This study seeks to understand which factors influence physician prescribing behavior regarding SUP. We designed a cross sectional web-based survey to assess physicians' knowledge, beliefs, and behavior surrounding the prescribing of SUP for non-ICU patients. The survey was emailed to internal medicine residents and hospitalists at a university-affiliated tertiary care hospital. Clinically relevant bivariable associations were examined in logistic regression to determine whether these associations remained after adjustment for potential confounding factors. Sixty-nine percent of physicians reported prescribing SUP to ≥25% of patients. In multivariable analyses, the following factors were associated with higher level of prescribing (≥25%) of SUP: fear of gastrointestinal bleeding (OR = 2.7, 95% CI 1.07, 7.28) and of the legal repercussions of not prescribing SUP (OR = 3.02, 95% CI 1.07, 8.56), whereas knowledge of SUP indications (OR = 0.39, 95% CI 0.20, 0.74) and concern about side effects (OR = 0.24, 95% CI 0.09, 0.61) were associated with low prescribing behavior. Level of training was not associated with prescribing rate. Less than half of respondents were able to identify a single side effect of proton pump inhibitor therapy. Fear of legal repercussions and ignorance of the side effects of acid suppressive therapy were strongly associated with inappropriate prescribing of SUP. Educating physicians about the adverse effects of acid suppression therapy and about existing national guidelines might reduce inappropriate prescribing.


Non-oxidative intercalation and exfoliation of graphite by Brønsted acids.
Graphite intercalation compounds are formed by inserting guest molecules or ions between sp(2)-bonded carbon layers. These compounds are interesting as synthetic metals and as precursors to graphene. For many decades it has been thought that graphite intercalation must involve host-guest charge transfer, resulting in partial oxidation, reduction or covalent modification of the graphene sheets. Here, we revisit this concept and show that graphite can be reversibly intercalated by non-oxidizing Brønsted acids (phosphoric, sulfuric, dichloroacetic and alkylsulfonic acids). The products are mixtures of graphite and first-stage intercalation compounds. X-ray photoelectron and vibrational spectra indicate that the graphene layers are not oxidized or reduced in the intercalation process. These observations are supported by density functional theory calculations, which indicate a dipolar interaction between the guest molecules and the polarizable graphene sheets. The intercalated graphites readily exfoliate in dimethylformamide to give suspensions of crystalline single- and few-layer graphene sheets.


High-intensity exercise and muscle glycogen availability in humans.
This study investigated the effects of muscle glycogen availability on performance and selected physiological and metabolic responses during high-intensity intermittent exercise. Seven male subjects completed a regimen of exercise and dietary intake (48 h) to either lower and keep low (LOW-CHO) or lower and then increase (HIGH-CHO) muscle glycogen stores, on two separate occasions at least a week apart. On each occasion the subjects completed a short-term (<10 min) and prolonged (>30 min) intermittent exercise (IEX) protocol, 24 h apart, which consisted of 6-s bouts of high-intensity exercise performed at 30-s intervals on a cycle ergometer. Glycogen concentration (mean +/- SEM) in m. vastus lateralis before both IEx(short) and IEx(long) was significantly lower following LOW-CHO [180 (14), 181 (17) mmol kg (dw)(-1)] compared with HIGH-CHO [397 (35), 540 (25) mmol kg (dw)(-1)]. In both IEx(short) and IEx(long), significantly less work was performed following LOW-CHO compared with HIGH-CHO. In IEx(long), the number of exercise bouts that could be completed at a pre-determined target exercise intensity increased by 265% from 111 (14) following LOW-CHO to 294 (29) following HIGH-CHO (P < 0.05). At the point of fatigue in IEx(long), glycogen concentration was significantly lower with the LOW-CHO compared with HIGH-CHO [58 (25) vs. 181 (46) mmol kg (dw)(-1), respectively]. The plasma concentrations of adrenaline and nor-adrenaline (in IEx(short) and IEx(long)), and FFAand glycerol (in IEx(long)), increased several-fold above resting values with both experimental conditions. Oxygen uptake during the exercise periods in IEx(long), approached 70% of Vo2max. These results suggest that muscle glycogen availability can affect performance during both short-term and more prolonged high-intensity intermittent exercise and that with repeated exercise periods as short as 6 s, there can be a relatively high aerobic contribution.


Induced ferromagnetism at BiFeO3/YBa2Cu3O7 interfaces.
Transition metal oxides (TMOs) exhibit many emergent phenomena ranging from high-temperature superconductivity and giant magnetoresistance to magnetism and ferroelectricity. In addition, when TMOs are interfaced with each other, new functionalities can arise, which are absent in individual components. Here, we report results from first-principles calculations on the magnetism at the BiFeO3/YBa2Cu3O7 interfaces. By comparing the total energy for various magnetic spin configurations inside BiFeO3, we are able to show that a metallic ferromagnetism is induced near the interface. We further develop an interface exchange-coupling model and place the extracted exchange coupling interaction strengths, from the first-principles calculations, into a resultant generic phase diagram. Our conclusion of interfacial ferromagnetism is confirmed by the presence of a hysteresis loop in field-dependent magnetization data. The emergence of interfacial ferromagnetism should have implications to electronic and transport properties.


Allicin Modifies the Composition and Function of the Gut Microbiota in Alcoholic Hepatic Steatosis Mice.
The intestinal microbiome plays an important role in the pathogenesis of liver diseases. Alcohol intake induces gut microbiota dysbiosis and alters its function. This study investigated the antibiotic effect of allicin in mice with hepatic steatosis. Male C57BL/6 mice were administered an ethanol diet supplemented with allicin (5 and 20 mg/(kg bw day)) for 4 weeks. Allicin modified the gut microbiota composition. Cecal microbiota exhibited a positive correlation with alcohol and hepatic triacylglycerol, but were suppressed with allicin. Ethanol diet with 5 mg of allicin induced a lower intestinal permeability compared to the ethanol diet alone. Allicin mediated the lipopolysaccharide (LPS)-CD14-toll-like receptor 4 (TLR4)-induced hepatic inflammation pathway by reducing LPS, CD14, TLR4, and pro-inflammatory cytokines-tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6. However, hepatic inflammation primarily resulted from alcohol toxicity rather than LPS production in the gut. The prediction of functional profiles from metagenomic 16S ribosomal RNA (rRNA) data revealed different functional profiles in each group. The predicted aldehyde dehydrogenase tended to increase in alcoholic mice administered allicin. The predicted LPS-related pathway and LPS biosynthesis protein results exhibited a similar trend as plasma LPS levels. Thus, alcohol and allicin intake shapes the gut microbiota and its functional profile and improves the CD14-TLR4 pathway to alleviate inflammation in the liver.


Transcriptome analysis reveals heterogeneity in the injury response of kidney transplants.
We studied the transcripts that are increased by stress and injury in mouse kidney transplants, focusing on transcripts increased in parenchymal cells-injury and repair-induced transcripts (IRITs). We compared four types of stressed kidneys: isografts, allografts, host kidneys of mice with isografts and nontransplant kidneys with ischemic acute tubular necrosis (ATN). After excluding transcripts associated with infiltrating cells and interferon-gamma-induced transcripts, we defined 790 IRITs in isografts. IRITs were remarkably heterogeneous in timing and mechanisms. Some were increased in host as well as donor kidneys, reflecting systemic influences (wounding, anesthetic). Most reflected local stress, resembling changes in ATN despite the lack of ATN histopathology. Mathematical decomposition of IRIT expression patterns confirmed heterogeneity, separating IRIT changes into component subsets, with an early peak (day 1) showing systemic effects and late peaks that resembled ATN, manifested Tgf-ss1 effects and recapitulated embryonic development. In allografts IRITs were initially similar to isografts but diverged due to allogeneic injury. The allospecific induction of IRITs was T-cell-dependent but perforin-granzyme-independent, compatible with delayed type hypersensitivity. The alloresponse strikingly and selectively increased the late IRITs but not the IRITs that peak early, indicating that rejection triggers parenchymal responses similar to those in ATN.


Brief Report: Relationship between non-verbal IQ and gender in autism.
It has been proposed that females at risk for autism are protected in some way, so that only those with the greatest genetic liability are affected. Consequently, affected male siblings of females with autism should be more impaired than affected male siblings of male probands. One hundred and ninety-four (194) families with a single child with autism (simplex, SPX) and 154 families with more than one child with autism (multiplex, MPX) were examined on measures of severity, including non-verbal IQ. Among SPX families, girls had lower IQ than boys, but no such differences were seen among MPX families. Similarly, the affected brothers of girls with autism were no different from affected brothers of male probands. These data suggest that MPX and SPX families differ with respect to the relationship between gender and IQ.


Age-related changes in ecosystem structure and function and effects on water and carbon exchange in ponderosa pine.
As forests age, their structure and productivity change, yet in some cases, annual rates of water loss remain unchanged. To identify mechanisms that might explain such observations, and to determine if widely different age classes of forests differ functionally, we examined young (Y, approximately 25 years), mature (M, approximately 90 years) and old (O, approximately 250 years) ponderosa pine (Pinus ponderosa Dougl. ex P. Laws.) stands growing in a drought-prone region of central Oregon. Although the stands differed in tree leaf area index (LAIT) (Y = 0.9, M = 2.8, O = 2.1), cumulative tree transpiration measured by sap flow did not differ substantially during the growing season (100-112 mm). Yet when water was readily available, transpiration per unit leaf area of the youngest trees was about three times that of M trees and five times that of O trees. These patterns resulted from a nearly sixfold difference in leaf specific conductance (KL) between the youngest and oldest trees. At the time of maximum transpiration in the Y stand in May-June, gross carbon uptake (gross ecosystem production, GEP) was similar for Y and O stands despite an almost twofold difference in stand leaf area index (LAIS). However, the higher rate of water use by Y trees was not sustainable in the drought-prone environment, and between spring and late summer, KL of Y trees declined fivefold compared with a nearly twofold decline for M trees and a < 30% reduction in O trees. Because the Y stand contained a significant shrub understory and more exposed soil, there was no appreciable difference in mean daily latent energy fluxes between the Y stand and the older stands as measured by the eddy-covariance technique. These patterns resulted in 60 to 85% higher seasonal GEP and 55 to 65% higher water-use efficiency at the M and O stands compared with the Y stand.


A single pulse of diffuse contaminants alters the size distribution of natural phytoplankton communities.
The presence of a multitude of bioactive organic pollutants collectively classified as pharmaceuticals and personal care products (PPCPs) in freshwaters is of concern, considering that ecological assessments of their potential impacts on natural systems are still scarce. In this field experiment we tested whether a single pulse exposure to a mixture of 12 pharmaceuticals and personal care products, which are commonly found in European inland waters, can influence the size distributions of natural lake phytoplankton communities. Size is one of the most influential determinants of community structure and functioning, particularly in planktonic communities and food webs. Using an in-situ microcosm approach, phytoplankton communities in two lakes with different nutrient levels (mesotrophic and eutrophic) were exposed to a concentration gradient of the PPCPs mixture at five levels. We tested whether sub-lethal PPCPs doses affect the scaling of organisms' abundances with their size, and the slope of these size spectra, which describe changes in the abundances of small relative to large phytoplankton. Our results showed that a large proportion (approximately 80%) of the dataset followed a power-law distribution, thus suggesting evidence of scale invariance of abundances, as expected in steady state ecosystems. PPCPs were however found to induce significant changes in the size spectra and community structure of natural phytoplankton assemblages. The two highest treatment levels of PPCPs were associated with decreased abundance of the most dominant size class (nano-phytoplankton: 2-5 μm), leading to a flattening of the size spectra slope. These results suggest that a pulse exposure to PPCPs induce changes that potentially lead to unsteady ecosystem states and cascading effects in the aquatic food webs, by favoring larger non-edible algae at the expense of small edible species. We propose higher susceptibility due to higher surface to volume ratio in small species as the likely cause of these structural changes.


Acute Infectious Gastroenteritis Potentiates a Crohn's Disease Pathobiont to Fuel Ongoing Inflammation in the Post-Infectious Period.
Crohn's disease (CD) is a chronic inflammatory condition of diverse etiology. Exposure to foodborne pathogens causing acute gastroenteritis produces a long-term risk of CD well into the post-infectious period but the mechanistic basis for this ongoing relationship to disease onset is unknown. We developed two novel models to study the comorbidity of acute gastroenteritis caused by Salmonella Typhimurium or Citrobacter rodentium in mice colonized with adherent-invasive Escherichia coli (AIEC), a bacterial pathobiont linked to CD. Here, we show that disease activity in the post-infectious period after gastroenteritis is driven by the tissue-associated expansion of the resident AIEC pathobiont, with an attendant increase in immunopathology, barrier defects, and delays in mucosal restitution following pathogen clearance. These features required AIEC resistance to host defense peptides and a fulminant inflammatory response to the enteric pathogen. Our results suggest that individuals colonized by AIEC at the time of acute infectious gastroenteritis may be at greater risk for CD onset. Importantly, our data identify AIEC as a tractable disease modifier, a finding that could be exploited in the development of therapeutic interventions following infectious gastroenteritis in at-risk individuals.


Nearly three orders of magnitude enhancement of Goos-Hanchen shift by exciting Bloch surface wave.
Goos-Hanchen effect is experimentally studied when the Bloch surface wave is excited in the forbidden band of a one-dimensional photonic band-gap structure. By tuning the refractive index of the cladding covering the truncated photonic crystal structure, either a guided or a surface mode can be excited. In the latter case, strong enhancement of the Goos-Hanchen shift induced by the Bloch-surface-wave results in sub-millimeter shifts of the reflected beam position. Such giant Goos-Hanchen shift, ~750 times of the wavelength, could enable many intriguing applications that had been less than feasible to implement before.


Cuprizone demyelination induces a unique inflammatory response in the subventricular zone.
Cuprizone leads to demyelination of the corpus callosum (CC) and activates progenitor cells in the adjacent subventricular zone (SVZ), a stem cell niche which contributes to remyelination. The healthy SVZ contains semi-activated microglia and constitutively expresses the pro-inflammatory molecule galectin-3 (Gal-3) suggesting the niche uniquely regulates inflammation. We studied the inflammatory response to cuprizone in the SVZ and CC in Gal-3 knockout mice using immunohistochemistry and with the in vitro neurosphere assay. Cuprizone caused loss of myelin basic protein (MBP) immunofluorescence in the CC suggesting demyelination. Cuprizone increased the density of CD45+/Iba1+ microglial cells and also increased Gal-3 expression in the CC. Surprisingly, the number of Gal-3+ and CD45+ cells decreased in the SVZ after cuprizone, suggesting inflammation was selectively reduced therein. Inflammation can regulate SVZ proliferation and indeed the number of phosphohistone H3+ (PHi3+) cells decreased in the SVZ but increased in the CC in both genotypes after cuprizone treatment. BrdU+ SVZ cell numbers also decreased in the SVZ after cuprizone, and this effect was significantly greater at 3 weeks in Gal-3 (-/-) mice compared to WT, suggesting Gal-3 normally limits SVZ cell emigration following cuprizone treatment. This study reveals a uniquely regulated inflammatory response in the SVZ and shows that Gal-3 participates in remyelination in the cuprizone model. This contrasts with more severe models of demyelination which induce SVZ inflammation and suggests the extent of demyelination affects the SVZ neurogenic response.


Aggregation of nanoparticles on one and two-component bilayer membranes.
Using dissipative particle dynamics simulations the aggregation of nanoparticles on single and two-component bilayers is investigated. For a uniform bilayer the aggregation of nanoparticles depends strongly on the location of the particles in the bilayer; particles residing on the bilayer exterior cluster strongly under the influence of bilayer-mediated interactions, whereas the interaction between the particles in the bilayer interior is significantly weaker leading to more loosely bound, dynamic aggregates. The aggregation of nanoparticles on two-component bilayers composed of immiscible components changes due to competition between nanoparticle clustering and their adsorption on the boundary between the bilayer components. This reduces the size of the nanoparticle clusters formed on the bilayer exterior, with the clusters adhering onto the boundary between the bilayer components. Due to their weaker attraction nanoparticles in the interior of a mixed bilayer no longer aggregate and instead form strings along the boundary between the two bilayer components. Nanoparticles with an affinity to one bilayer component nucleate small domains of their favoured component around themselves. For asymmetric mixtures this leads to a notable change in the aggregation behaviour of the nanoparticles.


Changes in oak (Quercus robur) photosynthesis after winter moth (Operophtera brumata) herbivory are not explained by changes in chemical or structural leaf traits.
Insect herbivores have the potential to change both physical and chemical traits of their host plant. Although the impacts of herbivores on their hosts have been widely studied, experiments assessing changes in multiple leaf traits or functions simultaneously are still rare. We experimentally tested whether herbivory by winter moth (Operophtera brumata) caterpillars and mechanical leaf wounding changed leaf mass per area, leaf area, leaf carbon and nitrogen content, and the concentrations of 27 polyphenol compounds on oak (Quercus robur) leaves. To investigate how potential changes in the studied traits affect leaf functioning, we related the traits to the rates of leaf photosynthesis and respiration. Overall, we did not detect any clear effects of herbivory or mechanical leaf damage on the chemical or physical leaf traits, despite clear effect of herbivory on photosynthesis. Rather, the trait variation was primarily driven by variation between individual trees. Only leaf nitrogen content and a subset of the studied polyphenol compounds correlated with photosynthesis and leaf respiration. Our results suggest that in our study system, abiotic conditions related to the growth location, variation between tree individuals, and seasonal trends in plant physiology are more important than herbivory in determining the distribution and composition of leaf chemical and structural traits.


SU-E-I-35: Experience with Th Zero Gravity Suit.
To compare the effectiveness of the Zero Gravity Suit (CFI Medical Solutions) in reducing the annual dose received by one physician in the Cardiac Catheterization Lab. A physician with a history of receiving high annual dose equivalents was purchased a Zero Gravity Suit. Film badges (Landauer) were worn at the collar and waist and monitored monthly. Previous dose history was acquired via the Landauer Exposure Report Archive. Annual values for Deep Dose Equivalent (DDE), Eye Dose Equivalent (LDE), and Shallow Dose Equivalent (SDE) were compared for the first full year of the Zero Gravity Suit (2011) to the previous four years (2007-2010). Readings for 2009 and 2007 were corrected for improper wear of the film badges. The reduction in dose equivalent for DDE ranged from 70%-87%. The reduction in dose equivalent for LDE ranged from 16%-60%. The reduction in dose equivalent for SDE ranged from 16%-60%. It was determined that the Zero Gravity Suit is effective at reducing the exposure of a physician with a history of high annual dose equivalents in our facilities Cardiac Catheterization Lab. While the suit did reduce the LDE and SDE it was more effective at reducing the DDE. During 2010 the suit was acquired midyear. The dose equivalent values would likely be similar to 2011 if the suit was introduced at the beginning of the year. In 2009 the physician was pregnant and attempting a reduced workload. The physician only uses the suit if a procedure is expected to take longer than fifteen minutes. The accuracy of the study is dependent upon the physician wearing the badges consistently and correctly and the physician using the Zero Gravity Suit consistently.


Parity and Risk of Thyroid Cancer: a Population-Based Study in Lithuania.
An association between parity and thyroid cancer risk has been investigated in a number of independent studies but yielded contradictory findings. The aim of this study was to explore the association between parity and thyroid cancer risk. The population-based cohort study in Lithuanian was conducted. The study dataset based on the linkages between all records from the 2001 population census, all cancer incidence records from the Lithuanian Cancer Registry, and all death and emigration records from Statistics Lithuania for the period between 6 April 2001 and 31 December 2009. Cox's proportional hazards regression models were used to estimate the hazard ratios (HRs) for parity, age at first birth, number of children, place of residence, education, and age at census. The cohort of 868,105 women was followed for 8.6 years, and 1775 thyroid cancer cases were diagnosed during the study period. The significantly higher thyroid cancer risk was observed among parous women (HR = 1.45, 95% CI: 1.20, 1.75) and in women with 1, 2, and 3 children, after adjusting for the possible confounding effects of relevant demographic variables. The findings of this study are consistent with the hypothesis that parity might be associated with the risk of thyroid cancer in women.


Expressing emotions, rejection sensitivity, and attachment in patients with fibromyalgia.
This study aimed to examine emotional awareness, control of emotions, and the childhood attachment process in fibromyalgia patients. The observational study was conducted with 117 participants (14 males, 103 females; mean age: 43.9±9.0 years; range, 22 to 64 years) between February 20, 2022, and May 20, 2022. Sixty-one patients and 56 healthy control subjects filled out a form including sociodemographic data, such as age, sex, occupation, and educational status. In addition, the participants answered the Expressing Emotions Scale, Rejection Sensitivity Scale, and Experiences in Close Relationships Scale. In our study, Expressing Emotions Scale scores are significantly higher in the healthy control group than in the patient group (p<0.05). Rejection Sensitivity Scale scores are significantly higher in the patient group than in the healthy control group (p<0.05). In the Experiences in Close Relationships Scale, a significant difference was determined between the patient and control groups in the avoidant attachment subdimension (p<0.05). Similarly, a significant difference was observed between both groups in terms of the anxiety attachment subdimension (p<0.05). The ability to express emotions is lower and avoidant and anxious attachment rates are higher in fibromyalgia patients.


Innovative tools help counselors discuss childhood obesity with parents.
Childhood overweight and obesity pose potential health risks for many children under the age of 5 years. Women, Infants, and Children (WIC) nutritionists are in a unique position to help reduce this problem because of their frequent counseling contacts with clients during certification visits. Therefore, four new tools to facilitate nutritional counseling of parents of overweight children during certifications were developed and systematically evaluated. The Nutrition and Activity Self-History (NASH) form, Report Card/Action Plan (ReCAP), Talking Tips, and Healthy Weight Poster were evaluated by WIC nutritionists via an online survey. Anchors on the Likert scale were 0 for Strongly Disagree to 6 for Strongly Agree. Four regional focus groups were also conducted. Data were analyzed descriptively. The response rate on the survey was 83% (n=63). Focus groups were comprised of staff that volunteered to participate (n=34). The NASH form, which replaces a food frequency questionnaire for identifying nutrition risk, had a mean rating of 5.20 as "Helpful when counseling about weight." The ReCAP, Talking Tips, and Healthy Weight Poster achieved mean ratings of 5.70, 4.75, and 5.30, respectively, in this category. Focus group responses were very positive about the usefulness of the ReCAP and Healthy Weight Poster to visually convey the concept of BMI percentile for age using a green, yellow, and red color-coded "traffic light" approach to showing healthy versus unhealthy BMI values. WIC programs and other pediatric health care settings may want to consider adopting these innovative tools to better serve their clients and address pediatric overweight in the populations they serve.


The effects of target template specificity on visual search in real-world scenes: evidence from eye movements.
We can locate an object more quickly in a real-world scene when a specific target template is held in visual working memory, but it is not known exactly how a target template's specificity affects real-world search. In the present study, we compared word and picture cues in real-world scene search. Using an eye-tracker, we segmented search time into three behaviorally defined epochs: search initiation time, scanning time, and verification time. Results from three experiments indicated that target template specificity affects scanning and verification time. Within the scanning epoch, target template specificity affected the number of scene regions visited and the mean fixation duration. Changes to SOA did not affect this pattern of results. Similarly, the pattern of results did not change when participants were familiarized with target images prior to testing, suggesting that an immediately preceding picture provides a more useful search template than one stored in long-term memory. The results suggest that the specificity of the target cue affects both the activation map representing potential target locations and the process that matches a fixated object to an internal representation of the target.


Efficacy and safety of namilumab, a human monoclonal antibody against granulocyte-macrophage colony-stimulating factor (GM-CSF) ligand in patients with rheumatoid arthritis (RA) with either an inadequate response to background methotrexate therapy or an inadequate response or intolerance to an anti-TNF (tumour necrosis factor) biologic therapy: a randomized, controlled trial.
Namilumab (AMG203), an immunoglobulin G1 monoclonal antibody that binds with high affinity to granulocyte-macrophage colony-stimulating factor (GM-CSF), was evaluated in a phase II randomized, double-blind, placebo-controlled study to investigate the efficacy and safety in patients with rheumatoid arthritis (RA) with an inadequate response to methotrexate (MTX-IR) or anti-tumour necrosis factor therapy (TNF-IR). Subcutaneous namilumab (20, 80, or 150 mg) or placebo was administered at baseline and weeks 2, 6, and 10 in patients on stable background methotrexate therapy who were with MTX-IR or TNF-IR. Primary endpoint was mean change from baseline in the 28-joint Disease Activity Score, C-reactive protein version (DAS28-CRP) at week 12 comparing each of the three doses of namilumab to placebo. Safety and tolerability were assessed by adverse events (AEs) and pulmonary parameters. Results were analysed using the per-protocol population. One hundred eight patients from Europe and Japan (48.4 ± 12.02 years old; 77.8% female; mean DAS28-CRP 5.60-5.79; rheumatoid factor/anti-citrullinated protein antibodies + 75%) were randomized to placebo or namilumab 20, 80, or 150 mg (n = 27, 28, 25, and 28, respectively). Ninety-two were MTX-IR; 16 were TNF-IR. At week 12, a statistically significant difference in DAS28-CRP (p = 0.005) was seen for namilumab 150 mg versus placebo and separation was seen as early as week 2 for namilumab 150 mg (p < 0.05), with higher ACR50 and response rates versus placebo at week 12. A dose-response effect was observed across the DAS28-CRP endpoint with separation versus placebo evident from week 2. The most common treatment-emergent AEs were nasopharyngitis (18.5%, 17.9%, 4.0%, 14.3%), dyspnoea (0.0%, 3.6%, 8.0%, 10.7%), bronchitis (7.4%, 3.6%, 4.0%, 3.6%), and headache (3.7%, 3.6%, 12.0%, 0.0%) for placebo and 20, 80, or 150 mg of namilumab, respectively. No serious infections were observed. One serious AE (myocardial infarction) was observed with 150 mg of namilumab. There was no apparent dose relationship for AEs. A biomarker-based disease activity score showed a dose-dependent decrease at week 12. This phase II study demonstrates the benefit of inhibiting macrophage activity targeting the GM-CSF for RA. The study met its primary endpoint with a clear dose-response effect. An acceptable tolerability profile was demonstrated over the 12-week study. ClinicalTrials.gov, NEXUS; NCT02379091 , submitted November 28, 2014.


Telomere-based proliferative lifespan barriers in Werner-syndrome fibroblasts involve both p53-dependent and p53-independent mechanisms.
Werner-syndrome fibroblasts have a reduced in vitro life span before entering replicative senescence. Although this has been thought to be causal in the accelerated ageing of this disease, controversy remains as to whether Werner syndrome is showing the acceleration of a normal cellular ageing mechanism or the occurrence of a novel Werner-syndrome-specific process. Here, we analyse the signalling pathways responsible for senescence in Werner-syndrome fibroblasts. Cultured Werner-syndrome (AG05229) fibroblasts senesced after approximately 20 population doublings with most of the cells having a 2N content of DNA. This was associated with hypophosphorylated pRb and high levels of p16(Ink4a) and p21(Waf1). Senescent AG05229 cells re-entered the cell cycle following microinjection of a p53-neutralizing antibody. Similarly, production of the human papilloma virus 16 E6 oncoprotein in presenescent AG05229 cells resulted in senescence being bypassed and extended cellular life span. Werner-syndrome fibroblasts expressing E6 did not proliferate indefinitely but reached a second proliferative lifespan barrier, termed M(int), that could be bypassed by forced production of telomerase in post-M1 E6-producing cells. The conclusions from these studies are that: (1) replicative senescence in Werner-syndrome fibroblasts is a telomere-induced p53-dependent event; and (2) the intermediate lifespan barrier M(int) is also a telomere-induced event, although it appears to be independent of p53. Werner-syndrome fibroblasts resemble normal human fibroblasts for both these proliferative lifespan barriers, with the strong similarity between the signalling pathway linking telomeres to cell-cycle arrest in Werner-syndrome and normal fibroblasts providing further support for the defect in Werner syndrome causing the acceleration of a normal ageing mechanism.


Ionizing radiation sensitizes breast cancer cells to Bcl-2 inhibitor, ABT-737, through regulating Mcl-1.
Breast-conserving surgery followed by radiation therapy has become the standard of care for early stage breast cancer. However, there are some patients that develop a local failure. We have previously shown that Bcl-2 overexpression was associated with an increased risk of local recurrence in patients with early stage breast cancer. The purpose of this study was to explore an approach to overcome radiation resistance by targeting pro-survival Bcl-2 family proteins in breast cancer cells. The breast cancer cell lines MCF-7, ZR-75-1 and MDA-MB231 were used in this study. siRNAs were employed to silence myeloid cell leukemia 1 (Mcl-1). A small molecule inhibitor of Bcl-2, ABT-737, was used to target anti-apoptotic Bcl-2 family proteins. Apoptosis was identified by FITC Annexin V, PI staining and Western blot analysis. The sensitivity to ionizing radiation and ABT-737 were measured by clonogenic assays. The effect of radiation and ABT-737 was also tested in a MCF-7 xenograft mouse model. Our data demonstrate that the combination of ABT-737 and radiation-induced apoptosis had an inhibitory effect on breast cancer cell proliferation. However, treatment with ABT-737 resulted in elevated Mcl-1 in breast cancer cell lines. Targeting Mcl-1 by siRNA sensitized MCF-7 cells to ABT-737. We revealed that radiation blunted Mcl-1 elevation induced by ABT-737, and that radiation downregulated Mcl-1 by promoting its degradation. Our results indicate that radiation and ABT-737 exert a synergistic effect on breast cancer cell lines through downregulating Mcl-1 and activating the bak-apoptotic pathway. These results support the combination of radiation and pro-survival Bcl-2 family inhibitor as a potential novel therapeutic strategy in the local-regional management of breast cancer.


Mutations of RNA polymerase II activate key genes of the nucleoside triphosphate biosynthetic pathways.
The yeast URA2 gene, encoding the rate-limiting enzyme of UTP biosynthesis, is transcriptionally activated by UTP shortage. In contrast to other genes of the UTP pathway, this activation is not governed by the Ppr1 activator. Moreover, it is not due to an increased recruitment of RNA polymerase II at the URA2 promoter, but to its much more effective progression beyond the URA2 mRNA start site(s). Regulatory mutants constitutively expressing URA2 resulted from cis-acting deletions upstream of the transcription initiator region, or from amino-acid replacements altering the RNA polymerase II Switch 1 loop domain, such as rpb1-L1397S. These two mutation classes allowed RNA polymerase to progress downstream of the URA2 mRNA start site(s). rpb1-L1397S had similar effects on IMD2 (IMP dehydrogenase) and URA8 (CTP synthase), and thus specifically activated the rate-limiting steps of UTP, GTP and CTP biosynthesis. These data suggest that the Switch 1 loop of RNA polymerase II, located at the downstream end of the transcription bubble, may operate as a specific sensor of the nucleoside triphosphates available for transcription.


Artificial Biosystem for Modulation of Interactions between Antigen-Presenting Cells and T Cells.
T cell activation is triggered by signal molecules on the surface of antigen-presenting cells (APC) and subsequent exertion of cellular forces. Deciphering the biomechanical and biochemical signals in this complex process is of interest and will contribute to an improvement in immunotherapy strategies. To address underlying questions, coculture and biomimetic models are established. Mature dendritic cells (mDC) are first treated with cytochalasin B (CytoB), a cytoskeletal disruption agent known to lower apparent cellular stiffness and reduction in T cell proliferation is observed. It is attempted to mimic mDC and T cell interactions using polyacrylamide (PA) gels with defined stiffness corresponding to mDC (0.2-25 kPa). Different ratios of anti-CD3 (aCD3) and anti-CD28 (aCD28) antibodies are immobilized onto PA gels. The results show T cell proliferation is triggered by both aCD3 and aCD28 in a stiffness-dependent manner. Cells cultured on aCD3 immobilized on gels has significantly enhanced proliferation and IL-2 secretion, compared to aCD28. Furthermore, ZAP70 phosphorylation is enhanced in stiffer substrate a in a aCD3-dependent manner. The biosystem provides an approach to study the reduction of T cell proliferation observed on CytoB-treated mDC. Overall, the biosystem allows distinguishing the impact of biophysical and biochemical signals of APC and T cell interactions in vitro.


Protocol for a systematic review of telephone delivered psychosocial interventions on relapse prevention, adherence to psychiatric medication and health risk behaviours in adults with a psychotic disorder.
The mental and physical health of individuals with a psychotic illness are typically poor. When adhered to, medication can reduce relapse. However, despite adherence, relapse remains common and functional outcomes often remain compromised. Compliance is also typically low. Cardiovascular-related morbidity and mortality is also elevated, along with several important modifiable health risk behaviours. Access to psychosocial interventions is therefore important, but currently limited. Telephone delivered interventions represent a promising solution, although further clarity is needed. Accordingly, we aim to provide an overview and critical analysis of the current state of evidence for telephone delivered psychosocial interventions targeting key health priorities in adults with a psychotic disorder, including (1) relapse, (2) adherence to psychiatric medication and/or (3) modifiable cardiovascular health risk behaviours. Our methods are informed by published guidelines. The review is registered and any protocol amendments will be tracked. Ten electronic peer-reviewed and four grey literature databases have been identified. Preliminary searches have been conducted for literature on psychosocial telephone interventions targeting relapse, medication adherence and/or health risk behaviours in adults with a psychotic disorder. Articles classified as 'evaluation' will be assessed against standardised criteria and checked by an independent assessor. The searches will be re-run just before final analyses and further studies retrieved for inclusion. A narrative synthesis will be reported, structured around intervention type and content, population characteristics and outcomes. Where possible, 'summary of findings' tables will be generated for each comparison. For the primary outcome of each trial, when data are available, we will calculate a risk ratio and its 95% CI (dichotomous outcomes) and/or effect size according to Cohen's formula (continuous outcomes). No ethical issues are foreseen. Findings will be disseminated widely to clinicians and researchers via journal publication and conference presentation(s). PROSPERO CRD42015025402.


Land use change effects on forest carbon cycling throughout the southern United States.
We modeled the effects of afforestation and deforestation on carbon cycling in forest floor and soil from 1900 to 2050 throughout 13 states in the southern United States. The model uses historical data on gross (two-way) transitions between forest, pasture, plowed agriculture, and urban lands along with equations describing changes in carbon over many decades for each type of land use change. Use of gross rather than net land use transition data is important because afforestation causes a gradual gain in carbon stocks for many decades, while deforestation causes a much more rapid loss in carbon stocks. In the South-Central region (Texas to Kentucky) land use changes caused a net emission of carbon before the 1980s, followed by a net sequestration of carbon subsequently. In the Southeast region (Florida to Virginia), there was net emission of carbon until the 1940s, again followed by net sequestration of carbon. These results could improve greenhouse gas inventories produced to meet reporting requirements under the United Nations Framework Convention on Climate Change. Specifically, from 1990 to 2004 for the entire 13-state study area, afforestation caused sequestration of 88 Tg C, and deforestation caused emission of 49 Tg C. However, the net effect of land use change on carbon stocks in soil and forest floor from 1990 to 2004 was about sixfold smaller than the net change in carbon stocks in trees on all forestland. Thus land use change effects and forest carbon cycling during this period are dominated by changes in tree carbon stocks.


On the role of mRNA secondary structure in bacterial translation.
Messenger RNA (mRNA) is no longer considered as a mere informational molecule whose sole function is to convey the genetic information specified by DNA to the ribosome. Beyond this primary function, mRNA also contains additional instructions that influence the way and the extent to which this message is translated by the ribosome into protein(s). Indeed, owing to its intrinsic propensity to quickly and dynamically fold and form higher order structures, mRNA exhibits a second layer of structural information specified by the sequence itself. Besides influencing transcription and mRNA stability, this additional information also affects translation, and more precisely the frequency of translation initiation, the choice of open reading frame by recoding, the elongation speed, and the folding of the nascent protein. Many studies in bacteria have shown that mRNA secondary structure participates to the rapid adaptation of these versatile organisms to changing environmental conditions by efficiently tuning translation in response to diverse signals, such as the presence of ligands, regulatory proteins, or small RNAs. This article is categorized under: Regulatory RNAs/RNAi/Riboswitches > Regulatory RNAs RNA Structure and Dynamics > Influence of RNA Structure in Biological Systems Translation > Translation Regulation.


Protective effect of melatonin and 1,25-dihydroxyvitamin D3 on renal ischemia-reperfusion injury in rats.
Ischemia-reperfusion (I/R) injury induces the generation of reactive oxygen species (ROS) which affect many organs. This study was designed to investigate the roles of melatonin and 1,25-dihydroxyvitamin D3 (VD3) on renal I/R injury. Thirty male Wistar albino rats were divided into five groups: group 1, control; group 2, right nephrectomy (RN) + I/R in the contralateral kidney; group 3, melatonin + RN + I/R; group 4, VD3 + RN + I/R; and group 5, melatonin + VD3 + RN + I/R. Melatonin (10 mg/kg), VD3 (0.5 μg/kg), and melatonin plus VD3 were injected intraperitoneally for 7 days before renal I/R. After 7 days, right nephrectomy was initially performed and left renal artery was clamped for 45 min. After 45-min reperfusion, the serum and kidney tissue samples were obtained for assays. Melatonin and VD3 had an ameliorative effect on biochemical parameters such as serum creatinine (SCr) and blood urea nitrogen (BUN). Renal tissue malondialdehyde (MDA), glutathione (GSH), nitric oxide (NO) levels, and superoxide dismutase (SOD) activity were determined. Renal I/R decreased the kidney tissue GSH levels and SOD activity and increased the NO levels as compared with control group. However, melatonin and VD3 and melatonin plus VD3 treatment significantly increased the tissue GSH levels and SOD activity and decreased the NO levels compared with those of I/R group. Meanwhile, MDA levels were not different between the control and I/R groups. But, MDA levels decreased in all treated groups compared to I/R and control groups. These data support that melatonin and VD3 have beneficial effects on renal injury.