Monoclonal anti-lambda 5 antibody FS1 identifies a 130 kDa protein associated with lambda 5 and Vpre-B on the surface of early pre-B cell lines.
mAbs specific for mouse lambda 5 protein were prepared by fusion of spleen cells from a hamster immunized with recombinant lambda 5 protein synthesized in bacteria and the mouse myeloma cell line SP2/0-Ag14. Here we report the characteristics of the antibodies produced by the FS1 hybridoma. FS1 antibody stains a variety of mouse pre-B cell lines but not B cell lines or T cell lines. The staining of the pre-B cell lines A-1 and C-7 by phycoerythrin (PE)-conjugated FS1 (FS1-PE) can be blocked by preincubation of these cells with unconjugated FS1 antibody or with affinity purified polyclonal lambda 5 specific Ig but not with normal hamster or mouse IgG or with affinity purified polyclonal anti-Mb-1 Ig. From these experiments we concluded that FS1 specifically recognizes lambda 5 protein. We used FS1-PE to probe for surface (s) lambda 5+ cells in normal BALB/c mouse bone marrow. Such cells were undetectable when total bone marrow or FACS sorted subpopulations were analyzed. However, when B220+, CD43+, s lambda 5-bone marrow cells were cultured for 4 days on the stromal cell line FLST2 in the presence of IL-7, s lambda 5 expression became apparent. Further expansion of these cells in IL7 alone augmented the s lambda 5 expression to readily detectable levels. This modulation may indicate that s lambda 5 expression on normal bone marrow cells in vivo is transient and that at any given moment only a small fraction of bone marrow cells expresses low levels of lambda 5 protein on the surface.(ABSTRACT TRUNCATED AT 250 WORDS)


The strategic role of recycling centres for environmental performance of waste management systems.
This paper analyses how different actors influence the sorting quality of waste at recycling centres. Users (i.e. citizens) play an essential role since they conduct the actual sorting. They have difficulties sorting many of their discarded products, leading to decreased performance of the entire waste management system of which recycling centres are a part. Several measures addressing this problem are identified such as product design, improved terminology for labelling waste and increased manning at recycling centres. A fundamental task for managers and employees is to further develop information and guidance for users, both at home and at recycling centres. Several obstacles for improvements are also discussed, including working conditions and the economy of recycling centres, as well as the routines for communication and quality assurance among actors in the recycling business.


Coexpression network and trans-activation analyses of maize reproductive phasiRNA loci.
The anther-enriched phased, small interfering RNAs (phasiRNAs) play vital roles in sustaining male fertility in grass species. Their long non-coding precursors are synthesized by RNA polymerase II and are likely regulated by transcription factors (TFs). A few putative transcriptional regulators of the 21- or 24-nucleotide phasiRNA loci (referred to as 21- or 24-PHAS loci) have been identified in maize (Zea mays), but whether any of the individual TFs or TF combinations suffice to activate any PHAS locus is unclear. Here, we identified the temporal gene coexpression networks (modules) associated with maize anther development, including two modules highly enriched for the 21- or 24-PHAS loci. Comparisons of these coexpression modules and gene sets dysregulated in several reported male sterile TF mutants provided insights into TF timing with regard to phasiRNA biogenesis, including antagonistic roles for OUTER CELL LAYER4 and MALE STERILE23. Trans-activation assays in maize protoplasts of individual TFs using bulk-protoplast RNA-sequencing showed that two of the TFs coexpressed with 21-PHAS loci could activate several 21-nucleotide phasiRNA pathway genes but not transcription of 21-PHAS loci. Screens for combinatorial activities of these TFs and, separately, the recently reported putative transcriptional regulators of 24-PHAS loci using single-cell (protoplast) RNA-sequencing, did not detect reproducible activation of either 21-PHAS or 24-PHAS loci. Collectively, our results suggest that the endogenous transcriptional machineries and/or chromatin states in the anthers are necessary to activate reproductive PHAS loci.


Neurological manifestations and implications of COVID-19 pandemic.
The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, China and rapidly spread worldwide, with a vast majority of confirmed cases presenting with respiratory symptoms. Potential neurological manifestations and their pathophysiological mechanisms have not been thoroughly established. In this narrative review, we sought to present the neurological manifestations associated with coronavirus disease 2019 (COVID-19). Case reports, case series, editorials, reviews, case-control and cohort studies were evaluated, and relevant information was abstracted. Various reports of neurological manifestations of previous coronavirus epidemics provide a roadmap regarding potential neurological complications of COVID-19, due to many shared characteristics between these viruses and SARS-CoV-2. Studies from the current pandemic are accumulating and report COVID-19 patients presenting with dizziness, headache, myalgias, hypogeusia and hyposmia, but also with more serious manifestations including polyneuropathy, myositis, cerebrovascular diseases, encephalitis and encephalopathy. However, discrimination between causal relationship and incidental comorbidity is often difficult. Severe COVID-19 shares common risk factors with cerebrovascular diseases, and it is currently unclear whether the infection per se represents an independent stroke risk factor. Regardless of any direct or indirect neurological manifestations, the COVID-19 pandemic has a huge impact on the management of neurological patients, whether infected or not. In particular, the majority of stroke services worldwide have been negatively influenced in terms of care delivery and fear to access healthcare services. The effect on healthcare quality in the field of other neurological diseases is additionally evaluated.


Purine nucleoside phosphorylase deficiency. Evidence for molecular heterogeneity in two families with enzyme-deficient members.
Purine-nucleoside phosphorylase (NP) deficiency is associated with severely defective thymus-derived (T)-cell and normally functioning bone marrow-derived (B)-cell immunity. In this study, two unrelated families with a total of three NP deficient members were investigated. High pressure liquid chromatography of the plasma of the three patients showed inosine levels greater than 66 muM. This nucleoside was absent from the plasma of their parents and control samples.NP was purified from normal human erythrocytes by affinity chromatography and an antiserum prepared in rabbits was used to study the NP variants in the two families. In family M the patient had no detectable erythrocyte NP activity and no detectable immunological-reacting material (irm) to the NP antibody. The parents, who are second cousins, had less than one-half of normal enzyme activity and approximately 14% irm attributable to a variant protein. Their electrophoretic patterns revealed a series of isozymes with slower than normal migration. In family B the patients had 0.5% residual enzyme activity and about one-half normal irm. Their electrophoretic pattern showed faintly staining bands which migrated faster than normal NP. The mother of the patients had one-half normal enzyme activity, 11% irm attributable to her variant protein, and a normal electrophoretic pattern. The father had less than one-half normal enzyme activity, equal amounts of normal and variant irm, and an electrophoretic pattern that showed increased activity of the more rapidly migrating isozyme bands.The combined use of immunological and electrophoretic techniques has shown the presence of three separate mutations; one in family M and two in family B associated with severely defective T-cell function.


Pregnancy Serum DLK1 Concentrations Are Associated With Indices of Insulin Resistance and Secretion.
Delta like noncanonical notch ligand 1 (DLK1) is a paternally expressed imprinted gene that encodes an epidermal growth factor repeat-containing transmembrane protein. A bioactive, truncated DLK1 protein is present in the circulation and has roles in development and metabolism. We sought to investigate links between maternal pregnancy circulating DLK1 concentrations and: (1) maternal and fetal DLK1 genotypes, (2) maternal insulin resistance and secretion, and (3) offspring size at birth. We measured third-trimester maternal serum DLK1 concentrations and examined their associations with parentally transmitted fetal and maternal DLK1 genotypes, indices of maternal insulin resistance and secretion derived from 75-g oral glucose tolerance tests performed around week 28 of pregnancy, and offspring size at birth in 613 pregnancies from the Cambridge Baby Growth Study. Maternal DLK1 concentrations were associated with the paternally transmitted fetal DLK1 rs12147008 allele (P = 7.8 × 10-3) but not with maternal rs12147008 genotype (P = 0.4). Maternal DLK1 concentrations were positively associated with maternal prepregnancy body mass index (P = 3.5 × 10-6), and (after adjustment for maternal body mass index) with both maternal fasting insulin resistance (Homeostatic Model Assessment of Insulin Resistance: P = 0.01) and measures of maternal insulin secretion in response to oral glucose (insulinogenic index: P = 1.2 × 10-3; insulin disposition index: P = 0.049). Further positive associations were found with offspring weight (P = 0.02) and head circumference at birth (P = 0.04). These results are consistent with a partial paternal or placental origin for the maternal circulating DLK1 which may lead to increased maternal circulating DLK1 concentrations, stimulation of maternal insulin resistance and compensatory hyperinsulinemia during pregnancy, and the promotion of fetal growth.


Startle activation is additive with voluntary cortical activation irrespective of stimulus modality.
When a startling acoustic stimulus (SAS) is presented during a simple reaction time (RT) task, it can trigger the prepared response through an involuntary initiation pathway. Previous research modelling the effects of presenting a SAS at various intervals following a non-startling auditory imperative signal (IS) suggested that involuntary initiation-related neural activation is additive with the voluntary initiation processes. The current study tested the predictions of this additive model when the SAS and IS are of different modalities by using a visual rather than auditory go-signal. Because voluntary RT latencies are delayed for visual stimuli compared to acoustic stimuli, it was hypothesised that the time course of additive activation would be similarly delayed. Participants performed 150 RT trials requiring a targeted 20° wrist extension task with a SAS presented 0-125 ms following a visual go-signal. Results were not different to those predicted by an additive model (p=0.979), yet were significantly different to those predicted by a horse-race model (p=0.037), indicating a joint contribution of voluntary and involuntary activation, even when the IS and SAS are of different modalities. Furthermore, the results indicated that voluntary RT differences due to stimulus modality are attributable to processes that occur prior to the increase in initiation-related activation.


A TFEB-Amplified Renal Cell Carcinoma with Long-Term, Complete Immunotherapy Response: Retrospective Support for the Value of Molecular Classification.
Recent years have seen the recognition and establishment of numerous subtypes of renal cell carcinoma (RCC), including adoption of an entire category of "molecularly defined renal carcinomas" in the fifth Edition of World Health Organization Classification. To add value, new diagnostic entities should be clinicopathologically distinct, or better, imply specific management and treatment angles, especially if adjunctive testing is needed for diagnosis. One such promising future treatment angle for a molecularly defined subtype, TFEB-amplified RCC, is immunotherapy, for which recent scholarship has demonstrated frequent expression of PD-L1. Herein, we report a case of metastatic TFEB-amplified RCC, where the patient experienced a long-term, complete response to PDL1-directed therapy, which had been serendipitously used years ago under a renal tumor subtype-agnostic indication. This promising experience suggests formal exploration of immunotherapy for these tumors.


Left ventricle geometry, atrial strain, ventricle strain, and hemodynamics across aortic valve before and after transcatheter aortic valve replacements.
Transcatheter aortic valve replacements (TAVRs) has become widespread throughout the world. To date, there are no echocardiographic studies of TAVR patients from Southeast Asia (SEA). We sought to evaluate (1) changes in echocardiographic and strain values pre- and post-TAVR (2) relationship between aortic stenosis (AS) severity and strain values, (3) left ventricle geometry in severe AS, (4) relationship of flow rate to dimensionless index (DVI) and acceleration time (AT), and (5) effect of strains on the outcome. Retrospective study of 112 TAVR patients in our centre from 2009 to 2020. The echocardiographic and strain images pre (within 1 month), post (day after), and 6 months post-TAVR were analyzed by expert echocardiographer. The ejection fraction (EF) increased at 6 months (53.02 ± 12.12% to 56.35 ± 9.00%) (p=0.044). Interventricular septal thickness in diastole (IVSd) decreased (1.27 ± 0.21 cm to 1.21 ± 0.23 cm) (p=0.038) and left ventricle internal dimension in diastole (LVIDd) decreased from 4.77 ± 0.64 cm to 4.49 ± 0.65 cm (p=0.001). No changes in stroke volume index (SVI pre vs 6 months p=0.187), but the flow rate increases (217.80 ± 57.61 mls/s to 251.94 ± 69.59 mls/s, p<0.001). Global longitudinal strain (GLS) improved from -11.44 ± 4.23% to -13.94 ± 3.72% (p<0.001), left atrial reservoir strain (Lar-S) increased from 17.44 ± 9.16% to 19.60 ± 8.77% (p=0.033). Eight patients (7.5%) had IVSd < 1.0 cm, and 4 patients (3.7%) had normal left ventricle (LV) geometry. There was linear relationship between IVSd and mean PG (r=0.208, p=0.031), between GLS to aortic valve area (AVA) and aortic valve area index (AVAi) (r = - 0.305, p=0.001 and r= - 0.316, p= 0.001). There was also relationship between AT (r=-0.20, p=0.04) and DVI (r=0.35, p<0.001) with flow rate. Patients who died late (after 6 months) had lower GLS at 6 months. (Alive; -13.94 ± 3.72% vs Died; -12.43 ± 4.19%, p=0.001). At 6 months, TAVR cause reverse remodelling of the LV with the reduction in IVSd, LVIDd, and improvement in GLS and LAr-S. There is a linear relationship between GLS and AVA and between IVSd and AVA.


Target-driven supramolecular self-assembly for selective amyloid-β photooxygenation against Alzheimer's disease.
Photo-oxygenation of β-amyloid (Aβ) has been considered an efficient way to inhibit Aβ aggregation in Alzheimer's disease (AD). However, current photosensitizers cannot simultaneously achieve enhanced blood-brain barrier (BBB) permeability and selective photooxygenation of Aβ, leading to poor therapeutic efficacy, severe off-target toxicity, and substandard bioavailability. Herein, an Aβ target-driven supramolecular self-assembly (PKNPs) with enhanced BBB penetrability and switchable photoactivity is designed and demonstrated to be effective in preventing Aβ aggregation in vivo. PKNPs are prepared by the self-assembly of the Aβ-targeting peptide KLVFF and an FDA-approved porphyrin derivative (5-(4-carboxyphenyl)-10,15,20-triphenylporphyrin). Due to the photothermal effect of PKNPs, the BBB permeability of PKNPs under irradiation is 8.5-fold higher than that of porphyrin alone. Moreover, upon selective interaction with Aβ, PKNPs undergo morphological change from the spherical to the amorphous form, resulting in a smart transformation from photothermal activity to photodynamic activity. Consequently, the disassembled PKNPs can selectively oxygenate Aβ without affecting off-target proteins (insulin, bovine serum albumin, and human serum albumin). The well-designed PKNPs exhibit not only improved BBB permeability but also highly selective Aβ photooxygenation. Furthermore, in vivo experiments demonstrate that PKNPs can alleviate Aβ-induced neurotoxicity and prolong the life span of the commonly used AD transgenic Caenorhabditis elegans CL2006. Our work may open a new path for using supramolecular self-assemblies as switchable phototheranostics for the selective and effective prevention of Aβ aggregation and related neurotoxicity in AD.


Acute hemorrhagic pancreatitis in mice: A study of glucoregulatory hormones and glucose metabolism.
In mice fed a choline-deficient diet containing 0.5% DL-ethionine (CDE) profound hypoglycemia develops, as do signs of shock, and the mice die of acute hemorrhagic pancreatitis (AHP). When fed a choline-supplemented diet containing 0.5% DL-ethionine (CSE), however, the mice have a chronic nonhemorrhagic pancreatitis with focal necrosis, do not show symptoms of shock, and survive. For investigation of the mechanism of hypoglycemia and the onset of shock in AHP, serum and pancreatic insulin and glucagon contents, serum glucose levels, and the morphologic characteristics of pancreatic islets were evaluated in mice fed laboratory Chow (LC); a choline-supplemented (CS) diet; a choline-deficient (CD) diet; or the CSE and CDE diets for 1, 2, or 3 days. The results indicate that onset of shock in animals with AHP may be due to hypoglycemia resulting from abnormal release of glucoregulatory hormones and their inability to maintain glucose output from liver and/or caused by active proteinases released from necrotic pancreas.


Uridine-5'-Triphosphate Partially Blocks Differentiation Signals and Favors a more Repair State in Cultured rat Schwann Cells.
Schwann cells (SCs) play a key role in peripheral nerve regeneration. After damage, they respond acquiring a repair phenotype that allows them to proliferate, migrate and redirect axonal growth. Previous studies have shown that Uridine-5'-Triphosphate (UTP) and its purinergic receptors participate in several pathophysiological responses in the nervous system. Our group has previously described how UTP induces the migration of a Schwannoma cell line and promotes wound healing. These data suggest that UTP participates in the signaling involved in the regeneration process. In the present study we evaluated UTP effects in isolated rat SCs and cocultures of SCs and dorsal root ganglia neurons. UTP reduced cAMP-dependent Krox-20 induction in SCs. UTP also reduced the N-cadherin re-expression that occurs when SCs and axons make contact. In myelinating cocultures, a non-significant tendency to a lower expression of P0 and MAG proteins in presence of UTP was observed. We also demonstrated that UTP induced SC migration without affecting cell proliferation. Interestingly, UTP was found to block neuregulin-induced phosphorylation of the ErbB3 receptor, a pathway involved in the regeneration process. These results indicate that UTP could acts as a brake to the differentiation signals, promoting a more migratory state in the repair-SCs.


[Diagnosis and surgical treatment of echinococcosis of the heart and pericardium].
Analysis of 9 observations of echinococcosis of the heart and 5 echinococcosis of the pericardium has shown the primary echinococcosis of the heart and pericardium to be a rare case (0,45%). Early death of parasites was observed in the heart followed by calcification and by self-healing. In patients presenting no complaints or objective disorders no operations are expedient. In a living cyst surgery (echinococcectomy) is indicated. With thinned walls of the heart operation should be accomplished by strengthening of the heart wall with pericardium or a diaphragm flap after B. V. Petrovski.


Activation of human THP-1 cells and rat bone marrow-derived macrophages by Helicobacter pylori lipopolysaccharide.
The mechanism by which Helicobacter pylori, which has little or no invasive activity, induces gastric-tissue inflammation and injury has not been well characterized. We have previously demonstrated that water-extracted proteins of H. pylori are capable of activating human monocytes by a lipopolysaccharide (LPS)-independent mechanism. We have now compared activation of macrophages by purified LPS from H. pylori and from Escherichia coli. LPS was prepared by phenol-water extraction from H. pylori 88-23 and from E. coli O55. THP-1, a human promyelomonocytic cell line, and macrophages derived from rat bone marrow each were incubated with the LPS preparations, and cell culture supernatants were assayed for production of tumor necrosis factor alpha (TNF-alpha), prostaglandin E2 (PGE2), and nitric oxide. THP-1 cells showed maximal activation by the LPS molecules after cell differentiation was induced by phorbol 12-myristate 13-acetate. Maximal TNF-alpha and PGE2 production occurred by 6 and 18 h, respectively, in both types of cells. In contrast, NO was produced by rat bone marrow-derived macrophages only and was maximal at 18 h. The minimum concentration of purified LPS required to induce TNF-alpha, PGE2, and NO responses in both types of cells was 2,000- to 30,000-fold higher for H. pylori than for E. coli. Purified LPS from three other H. pylori strains with different polysaccharide side chain lengths showed a similarly low level of activity, and polymyxin B treatment markedly reduced activity as well, suggesting that activation was a lipid A phenomenon. These results indicate the low biological activity of H. pylori LPS in mediating macrophage activation.


Long-term quality-of-life evaluation after head and neck cancer treatment in a developing country.
To evaluate the long-term quality of life of patients treated for head and neck cancer at a single institution in a developing country. Cross-sectional analysis of a consecutive series of patients. Tertiary cancer center hospital in Brazil. Eligible subjects included patients treated between 1974 and 1999 for head and neck carcinoma who had a minimum disease-free survival of 2 years and who completed a Portuguese version of the University of Washington Quality of Life (UW-QOL) questionnaire. Descriptive analyses of the results and comparisons of the scores for each UW-QOL domain, stratified by tumor site, were performed using nonparametric tests. Findings from 344 patients were analyzed. Of the study population, 140 (41%) had survived 2 to 5 years, 125 (36%) had survived 5 to 10 years, and 79 (23%) had survived more than 10 years since treatment. Primary tumor sites were in the oral cavity in 43.3% of cases, the oropharynx in 20.9%, the larynx in 32.0%, and the hypopharynx in 3.8%. In terms of treatment, 33.1% underwent surgery alone; 16.9%, radiotherapy alone; and 50% underwent combined treatment. Overall, 78.5% of the patients classified their own health as good or excellent. Stratified analysis showed that impairment in chewing and swallowing was more common in patients with oral and oropharyngeal tumors than in those with larynx and hypopharynx tumors, and speech impairment was more frequently related to patients with larynx and hypopharynx tumors than to those with oral and oropharynx tumors. In all tumor sites, the composite scores were significantly worse in advanced tumors than early stage tumors, but the use of combined treatment had the greatest negative impact on quality-of-life scores, after we adjusted for T and N stage with multivariable analyses (P<.001). The Portuguese version of the UW-QOL questionnaire was an effective tool to evaluate quality of life in a Brazilian population. Although many patients reported some limitations, most reported a good to excellent long-term quality of life.


Expression of a 95 kDa membrane protein is associated with low daunorubicin accumulation in leukaemic blast cells.
A 95 kDa membrane protein (P-95) has been previously noted to be overexpressed in a doxorubicin-resistant subline of the MCF-7 breast cancer line and in clinical samples obtained from patients with solid tumours refractory to doxorubicin. We performed Western blotting on blast cell lysates from adults with acute myeloid leukaemia, using antisera to P-95. Concomitant flow cytometric assays measured daunorubicin accumulation and retention. Blasts from 16/46 patient samples had detectable P-95 and had reduced accumulation of daunorubicin compared with the negative marrows. Experiments with the P-95 positive MCF-7 multidrug-resistant subline demonstrated decreased daunorubicin accumulation and retention relative to the sensitive parent line. AML blast cells positive for P-95 also demonstrated greater overall in vitro survival in the presence of daunorubicin relative to the P-95-negative samples. The expression of P-95 did not correlate with failure to achieve an initial complete remission with daunorubicin and cytarabine induction chemotherapy. We conclude that the P-95 protein may possess an efflux transporter function, and may represent another mechanism responsible for anthracycline resistance in acute myeloid leukaemia.


Solubility of proteins.
Solubility is a fundamental protein property that has important connotations for therapeutics and use in diagnosis. Solubility of many proteins is low and affect heterologous overexpression of proteins, formulation of products and their stability. Two processes are related to soluble and solid phase relations. Solubility refers to the process where proteins have correctly folded structure, whereas aggregation is related to the formation of fibrils, oligomers or amorphous particles. Both processes are related to some diseases. Amyloid fibril formation is one of the characteristic features in several neurodegenerative diseases, but it is related to many other diseases, including cancers. Severe complex V deficiency and cataract are examples of diseases due to reduced protein solubility. Methods and approaches are described for prediction of protein solubility and aggregation, as well as predictions of consequences of amino acid substitutions. Finally, protein engineering solutions are discussed. Protein solubility can be increased, although such alterations are relatively rare and can lead to trade-off with some other properties. The aggregation prediction methods mainly aim to detect aggregation-prone sequence patches and then making them more soluble. The solubility predictors utilize a wide spectrum of features.


Degradation of myofibrillar proteins by cathepsins B and D.
1. The procedure of Barrett [(1973) Biochem. J.131, 809-822] for isolating cathepsins B and D from human liver was modified for use with rat liver and skeletal muscle. The purified enzymes appeared to be similar to those reported in other species. 2. Sephadex G-75 chromatography of concentrated muscle extract resolved two peaks of cathepsin B inhibitory activity, corresponding to molecular weights of 12500 and 62000. 3. The degradation of purified myofibrillar proteins by cathepsins B and D was clearly demonstrated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. After incubation with enzyme, the polypeptide bands representing the substrates decreased in intensity and lower molecular weight products appeared. 4. Cathepsins B and D, purified from either rat liver or skeletal muscle, were shown to degrade myosin, purified from either rabbit or rat muscle. Soluble denatured myosin was degraded more extensively than insoluble native myosin. Degradation by cathepsin B was inhibited by lack of reducing agent, or by myoglobin, iodoacetic acid and leupeptin, but not by pepstatin. The same potential modifiers were applied to cathepsin D, and only pepstatin produced inhibition. 5. Rat liver cathepsin B had a pH optimum of 5.2 on native rabbit myosin. The pH optimum of cathepsin D was 4.0, with a shoulder of activity about 1pH unit above the optimum. 6. Rat liver cathepsins B and D were demonstrated to degrade rabbit F-actin at pH5.0, and were inhibited by leupeptin and pepstain, respectively. 7. The degradation of myosin and actin by cathepsin D was more extensive than that by cathepsin B.


Confirmation Bias in the Course of Instructed Reinforcement Learning in Schizophrenia-Spectrum Disorders.
A large body of research attributes learning deficits in schizophrenia (SZ) to the systems involved in value representation (prefrontal cortex, PFC) and reinforcement learning (basal ganglia, BG) as well as to the compromised connectivity of these regions. In this study, we employed learning tasks hypothesized to probe the function and interaction of the PFC and BG in patients with SZ-spectrum disorders in comparison to healthy control (HC) subjects. In the Instructed Probabilistic Selection task (IPST), participants received false instruction about one of the stimuli used in the course of probabilistic learning which creates confirmation bias, whereby the instructed stimulus is overvalued in comparison to its real experienced value. The IPST was administered to 102 patients with SZ and 120 HC subjects. We have shown that SZ patients and HC subjects were equally influenced by false instruction in reinforcement learning (RL) probabilistic task (IPST) (p-value = 0.441); however, HC subjects had significantly higher learning rates associated with the process of overcoming cognitive bias in comparison to SZ patients (p-value = 0.018). The behavioral results of our study could be hypothesized to provide further evidence for impairments in the SZ-BG circuitry; however, this should be verified by neurofunctional imaging studies.


The time course of attention: selection is transient.
The time course of attention has often been investigated using a spatial cuing task. However, attention likely consists of multiple components, such as selectivity (resolving competition) and orienting (spatial shifting). Here we sought to investigate the time course of the selective aspect of attention, using a cuing task that did not require spatial shifting. In several experiments, targets were always presented at central fixation, and were preceded by a cue at different cue-target intervals. The selection component of attention was investigated by manipulating the presence of distractors. Regardless of the presence of distractors, an initial rapid performance enhancement was found that reached its maximum at around 100 ms post cue onset. Subsequently, when the target was the only item in the display, performance was sustained, but when the target was accompanied by irrelevant distractor items, performance declined. This temporal pattern matches closely with the transient attention response that has been found in spatial cuing studies, and shows that the selectivity aspect of attention is transient.


The effect of calcium phosphate-containing desensitizing agent on the microtensile bond strength of multimode adhesive agent.
The aim of this study was to investigate the effect of calcium phosphate containing desensitizing pretreatments on the microtensile bond strength (MTBS) and microleakage of the multimode adhesive agent to dentin. In this study, twelve noncarious, freshly extracted human third molar teeth for MTBS and 20 premolar teeth for the microleakage test were used. The teeth were restored using Clearfil Universal Bond + Clearfil APX and Teeth mate Desensitizer (TMD). For MTBS test, Group 1: Self-etch, Group 2: Etch and rinse (G1 and 2, nondesensitizer treatment served as a control), Group 3: TMD/self-etch, Group 4: Acid-etch/TMD/etch and rinse. For microleakage test, Class V adhesive cavities (3 mm × 2 mm × 2 mm) were prepared and restored as mentioned before. The restored teeth were subjected to thermal cycling. The MTBS test was performed in all procedures. The MTBS data were submitted to a one-way ANOVA and post hoc Tukey test (P P Results: Control groups exhibited a higher mean MTBS value than TMD groups, and there were statistical differences between the groups. TMD groups also demonstrated significantly less microleakage than control groups (P Conclusions: This study proves that the application of TMD with a multimode adhesive bonding system produced significantly lower MTBS and microleakage.


Beyond single syllables: large-scale modeling of reading aloud with the Connectionist Dual Process (CDP++) model.
Most words in English have more than one syllable, yet the most influential computational models of reading aloud are restricted to processing monosyllabic words. Here, we present CDP++, a new version of the Connectionist Dual Process model (Perry, Ziegler, & Zorzi, 2007). CDP++ is able to simulate the reading aloud of mono- and disyllabic words and nonwords, and learns to assign stress in exactly the same way as it learns to associate graphemes with phonemes. CDP++ is able to simulate the monosyllabic benchmark effects its predecessor could, and therefore shows full backwards compatibility. CDP++ also accounts for a number of novel effects specific to disyllabic words, including the effects of stress regularity and syllable number. In terms of database performance, CDP++ accounts for over 49% of the reaction time variance on items selected from the English Lexicon Project, a very large database of several thousand of words. With its lexicon of over 32,000 words, CDP++ is therefore a notable example of the successful scaling-up of a connectionist model to a size that more realistically approximates the human lexical system.


Homeostatic generation of reactive oxygen species protects the zebrafish liver from steatosis.
Nonalcoholic fatty liver disease is the most common liver disease in both adults and children. The earliest stage of this disease is hepatic steatosis, in which triglycerides are deposited as cytoplasmic lipid droplets in hepatocytes. Through a forward genetic approach in zebrafish, we found that guanosine monophosphate (GMP) synthetase mutant larvae develop hepatic steatosis. We further demonstrate that activity of the small GTPase Rac1 and Rac1-mediated production of reactive oxygen species (ROS) are down-regulated in GMP synthetase mutant larvae. Inhibition of Rac1 activity or ROS production in wild-type larvae by small molecule inhibitors was sufficient to induce hepatic steatosis. More conclusively, treating larvae with hydrogen peroxide, a diffusible ROS that has been implicated as a signaling molecule, alleviated hepatic steatosis in both GMP synthetase mutant and Rac1 inhibitor-treated larvae, indicating that homeostatic production of ROS is required to prevent hepatic steatosis. We further found that ROS positively regulate the expression of the triglyceride hydrolase gene, which is responsible for the mobilization of stored triglycerides in hepatocytes. Consistently, inhibition of triglyceride hydrolase activity in wild-type larvae by a small molecule inhibitor was sufficient to induce hepatic steatosis. De novo GMP synthesis influences the activation of the small GTPase Rac1, which controls hepatic lipid dynamics through ROS-mediated regulation of triglyceride hydrolase expression in hepatocytes.


Sugarcane borer (Lepidoptera: Crambidae) resistance to transgenic Bacillus thuringiensis maize.
Transgenic maize, Zea mays L., expressing the Bacillus thuringiensis (Bt) CrylAb toxin has been planted to extensive areas across the United States and several other countries, but no resistance has been documented in field populations oflepidopteran target pests. This article describes the first report of resistance alleles to commercially available Cry1Ab Bt maize in a Louisiana population of sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae). Two hundred thirteen two-parent isolines of D. saccharalis were screened for Cry1Ab resistance on Bt maize leaf tissue using an F2 screening technique. Larvae representing three isolines survived >15 d on Bt tissue in the F2 generation. The second generation backcross progeny (B1F2) derived from isoline 52 completed larval development on Bt maize in the greenhouse. Segregation and resistance frequency analysis associated with isoline 52 suggested that Bt resistance is probably determined by a nearly completely recessive allele at a single locus. With this assumption, the estimated resistance allele frequency in this population is 0.0023, within a 95% confidence interval of 0.0003-0.0064.


HSPBs: small proteins with big implications in human disease.
Although initially somewhat ignored, recent studies have now clearly established that the diverse members of the human family of small HSPs (HSPB1-HSPB10) play crucial roles in a wide range of cell types to maintain the integrity and function of tissues, in particular that of nervous and muscular tissue. The 10 human HSPBs clearly have overlapping and non-overlapping functional characteristics. Their ability to self- and hetero-oligomerise provides the cells with a large array of potentially different, specific functions. Single HSPB members can have a multitude of functions (moonlighting) and act on different "clients", thus affecting a wide range of different processes or structures that can ultimately affect the rate of aging of tissues and entire organisms. This is underscored by the findings that some inherited diseases involve mutations in several HSPB members that cause premature (mostly muscle and neuronal) tissue degeneration. Inversely, cancer cell resistance to different anticancer therapies is associated with elevated expression of several HSPBs. Still, many unanswered questions exist about the precise functioning of HSPBs, their collaboration with other HSPB members as well as their functions within the entire cellular chaperone network. Also, better insight in the regulation of expression of the various members and how their function is modulated post-translationally is needed. Such may be crucially important to develop means to intervene with their function for therapeutic purposes, which would require functional down-regulation in cancer but up-regulation in, for instance, cardiac or degenerative neuro/neuromuscular diseases. This article is part of a Directed Issue entitled: Small HSPs in physiology and pathology.


Leveraging bimodal kinetics to improve detection specificity.
Optical microcavities are high sensitivity transducers able to detect single nanoparticles and molecules. However, the specificity of detection is dependent on the availability of an appropriate targeting moiety with minimal cross-reactivity. In the present work, an alternative approach is shown. Namely, using biotin-functionalized toroidal microcavities, the dissociation constant of biotin to two different streptavidin complexes (free and polystyrene bead) is determined. Based on the difference in affinity and in mass transport, the two complexes are identified from a mixture. By leveraging information in the binding site, improved specificity can be achieved.


Contribution of pancreatic edema and short-term ischemia to experimental acute pancreatitis in the rat. I. Procedure and pathomorphological investigations.
For experimental analysis of pathogenesis of acute pancreatitis a suitable animal model is necessary. The pancreatic (juice) edema and an alteration of cellular energy metabolism are important pathogenetic factors. A partial intrapancreatic edema was induced by stimulating secretion against permanent or temporary obstruction by ductal ligation. Short-term ischemia was used as tool for alteration of cellular energy metabolism. 24 h postoperatively morphological changes were macro- and microscopically investigated. The data show a correlation between duration of ischemia (10-30 min) and the frequency of acute pancreatitis at preexisting juice edema (44-68 percent). With increasing ischemia the morphological alterations (e.g.: necroses of adipose tissue) were enhanced, too. A hemorrhagic necrotizing pancreatitis was rarely observed. In comparison to other experimental models the presented one is characterized by a mostly moderate and light course of acute pancreatitis offering a good tool for elucidation of the very first stages of the pathogenesis.


[Risk factors in ischemic heart disease (arterial hypertension and hypercholesterolemia) and population age].
Comparative data of a 40% randomized screening of the male population in Moscow aged 50-59 (2000 males), a 5% randomized sample (392 males) aged 40-49, as well as 1631 males aged 40-49, subjected to prophylactic examinations are presented. The changes of the analyzed risk factors proved to be more pronounced in subjects aged 50-59, while significance of individual risk factors as regards the prevalence of ischemic heart disease (IHD) decreases with age. The subjects undergoing prophylactic examination represent a more risk population group in respect to the prevalence of IHD risk factors as compared with the randomized sample.


Progesterone inhibits insulin-like growth factor binding protein-1 (IGFBP-1) production by explants of the Fallopian tube.
The Fallopian tube provides the environment for early embryo growth, a process which is influenced by insulin-like growth factors (IGFs) in the tubal fluid. Whether the bioavailability of tubal IGFs is modulated by locally produced IGF-binding protein (IGFBP-1) is not clear. An explant culture system from human Fallopian tube mucosa was, therefore, developed enabling the potential for IGFBP-1 production by this tissue to be examined directly. Initial characterization of the system established that the explants maintained responsiveness to steroids. Thus, oviduct-specific glycoprotein production, a major product of the oviduct in vivo, continued to be made via an estrogen-sensitive pathway in the culture. The presence of mRNA for IGFBP-1 was established within the explants by the use of quantitative RT-PCR and IGFBP-1 protein was measured by enzyme-linked immunosorbent assay. Although insulin and estradiol had no consistent effect on IGFBP-1, addition of progesterone had a significant inhibitory effect on IGFBP-1 production, both at the mRNA and protein levels. A dose-range of progesterone revealed an incremental inhibitory effect of progesterone on IGFBP-1 output (maximal effect, 25-50 nmol/l), consistent with physiological inhibition of this process during the luteal phase. We suggest that progesterone might, therefore, play a role in controlling the bioavailability of IGFs to the embryo during early development within the Fallopian tube.


Novel application of a multiscale entropy index as a sensitive tool for detecting subtle vascular abnormalities in the aged and diabetic.
Although previous studies have shown the successful use of pressure-induced reactive hyperemia as a tool for the assessment of endothelial function, its sensitivity remains questionable. This study aims to investigate the feasibility and sensitivity of a novel multiscale entropy index (MEI) in detecting subtle vascular abnormalities in healthy and diabetic subjects. Basic anthropometric and hemodynamic parameters, serum lipid profiles, and glycosylated hemoglobin levels were recorded. Arterial pulse wave signals were acquired from the wrist with an air pressure sensing system (APSS), followed by MEI and dilatation index (DI) analyses. MEI succeeded in detecting significant differences among the four groups of subjects: healthy young individuals, healthy middle-aged or elderly individuals, well-controlled diabetic individuals, and poorly controlled diabetic individuals. A reduction in multiscale entropy reflected age- and diabetes-related vascular changes and may serve as a more sensitive indicator of subtle vascular abnormalities compared with DI in the setting of diabetes.


Computational screening of transition metal-doped phthalocyanine monolayers for oxygen evolution and reduction.
Rationally designing efficient, low-cost and stable catalysts toward the oxygen evolution reaction (OER) and the oxygen reduction reaction (ORR) is of significant importance to the development of renewable energy technologies. In this work, we have systematically investigated a series of potentially efficient and stable single late transition metal atom doped phthalocyanines (TM@Pcs, TM = Mn, Fe, Co, Ni, Cu, Ru, Rh, Pd, Ir and Pt) as single-atom catalysts (SACs) for applications toward the OER and ORR through a computational screening approach. Our calculations indicate that TM atoms can tightly bind with Pc monolayers with high diffusion energy barriers to prevent the isolated atoms from clustering. The interaction strength between intermediates and TM@Pc governs the catalytic activities for the OER and ORR. Among all the considered TM@Pc catalysts, Ir@Pc and Rh@Pc were found to be efficient OER electrocatalysts with overpotentials η OER of 0.41 and 0.44 V, respectively, and for the ORR, Rh@Pc exhibits the lowest overpotential η ORR of 0.44 V followed by Ir@Pc (0.55 V), suggesting that Rh@Pc is an efficient bifunctional catalyst for both the OER and ORR. Moreover, it is worth noting that the Rh@Pc catalyst can remain stable against dissolution under the pH = 0 condition. Ab initio molecular dynamic calculations suggest that Rh@Pc could remain stable at 300 K. Our findings highlight a novel family of two-dimensional (2D) materials as efficient and stable SACs and offer a new strategy for catalyst design.


A mobile phone-based retinal camera for portable wide field imaging.
Digital fundus imaging is used extensively in the diagnosis, monitoring and management of many retinal diseases. Access to fundus photography is often limited by patient morbidity, high equipment cost and shortage of trained personnel. Advancements in telemedicine methods and the development of portable fundus cameras have increased the accessibility of retinal imaging, but most of these approaches rely on separate computers for viewing and transmission of fundus images. We describe a novel portable handheld smartphone-based retinal camera capable of capturing high-quality, wide field fundus images. The use of the mobile phone platform creates a fully embedded system capable of acquisition, storage and analysis of fundus images that can be directly transmitted from the phone via the wireless telecommunication system for remote evaluation.


Fracture experience of patients with coeliac disease: a population based survey.
While coeliac disease is now recognised as being associated with both osteoporosis and osteomalacia, the size of any increase in the risk of fracture in patients with coeliac disease compared with the general population has not been quantified. To examine the fracture experience of adults with coeliac disease compared with the general population. Patients with coeliac disease diagnosed in adulthood and born before 1950, selected from two large population based disease registers, and age and sex frequency matched controls identified from local general practitioner lists. A four page lifestyle and general health questionnaire which included specific questions about fracture experience. Analysis was performed on 244 patients with coeliac disease and 161 controls, giving response rates of 89% and 72%, respectively. Eighty two (35%) coeliac patients and 53 (33%) controls reported ever having sustained one or more fractures, giving an age and sex adjusted odds ratio of 1.05 (95% confidence interval (CI) 0.68-1.62). The most common fracture site reported was the forearm or wrist, with an adjusted odds ratio of 1.21 (95% CI 0.66-2.25) for patients with coeliac disease having had a forearm or wrist fracture. Low trauma fractures were reported by 37 patients with coeliac disease (15.7%) and by 21 controls (13.8%), with an adjusted odds ratio of 1.16 (95% CI 0.65-2.10). The risk of low trauma fracture was slightly higher in coeliac men than women (odds ratio 1.28 compared with 1.12), but this difference was not statistically significant (p=0.84). After adjustment for age, sex, body mass index, and smoking status, patients with coeliac disease reported 13% more low trauma fractures than controls (odds ratio 1.13, 95% CI 0.60-2.12). There was no difference in low trauma fracture risk before and after diagnosis of coeliac disease. No overall increased fracture risk in patients with coeliac disease was observed. Although severe osteoporosis may develop in a subset of patients, as a whole patients with coeliac disease do not represent a population at particularly high risk of osteoporotic fracture and thus targeting them for osteoporosis screening and treatment is not justified.


EUROVISCO Good Practice Recommendations for a First Viscosupplementation in Patients with Knee Osteoarthritis.
Viscosupplementation (VS) with hyaluronic acid is widely used in the management of knee osteoarthritis. There is no clear recommendation on the decision-making to achieve VS. Based on extensive research of the literature and expert opinion, the members of the EUROVISCO (European Viscosupplementation Consensus Group) task force were asked to give their degree of agreement with 60 issues, using a Delphi method. The expert panel achieved unanimous agreement in favor of the following statements: It is recommended to assess pain on a visual or 10-point numeric scale before considering VS. VS can be considered for patients with pain scores between 3 and 8. A standard x-ray must be obtained before the decision of VS. If the x-ray is normal, osteoarthritis must be confirmed by MRI or computed tomography (CT) arthrogram before considering VS. The aims of VS are relieving pain, improving function, and reducing non-steroidal anti-inflammatory drug (NSAID) consumption. The use of VS must not be considered for treating an osteoarthritis flare. VS can be envisaged as a first-line pharmacological treatment in patients having a contra-indication to NSAIDs or analgesics. VS can be considered in patients with contra-indications to arthroplasty. In the case of severe comorbidities (diabetes, hypertension, gastrointestinal disorders, renal failure), VS can avoid the use of potentially dangerous treatments. VS can be considered in patients receiving antiplatelet agents, vitamin K antagonists, and direct factor Xa or thrombin inhibitors. Five other statements obtained a high level of consensus. These recommendations, illustrated in a decision algorithm, have been established to help practitioners in the decision-making of knee VS.


Resistance to 6-Methylpurine is Conferred by Defective Adenine Phosphoribosyltransferase in Tetrahymena.
6-methylpurine (6mp) is a toxic analog of adenine that inhibits RNA and protein synthesis and interferes with adenine salvage mediated by adenine phosphoribosyltransferase (APRTase). Mutants of the ciliated protist Tetrahymena thermophila that are resistant to 6mp were isolated in 1974, but the mechanism of resistance has remained unknown. To investigate 6mp resistance in T. thermophila, we created 6mp-resistant strains and identified a mutation in the APRTase genomic locus (APRT1) that is responsible for 6mp resistance. While overexpression of the mutated APRT1 allele in 6mp-sensitive cells did not confer resistance to 6mp, reduced wild-type APRT1 expression resulted in a significant decrease in sensitivity to 6mp. Knocking out or reducing the expression of APRT1 by RNA interference (RNAi) did not affect robust cell growth, which indicates that adenine salvage is redundant or that de novo synthesis pathways provide sufficient adenosine monophosphate for viability. We also explored whether 6mp resistance could be used as a novel inducible selection marker by generating 6mp- and paromomycin-resistant double mutants. While 6mp- and paromomycin-resistant double mutants did express fluorescent proteins in an RNAi-based system, the system requires optimization before 6mp resistance can be used as an effective inducible selection marker.


Nano Packaged Tamoxifen and Curcumin; Effective Formulation against Sensitive and Resistant MCF-7 Cells.
Tamoxifen is routinely used for treatment of Estrogen-positive breast carcinoma. Approximately, 50% of patients with metastatic cancer will develop resistance to Tamoxifen. In this research, Tamoxifen was combined with the anti-cancer compound Curcumin. Diblocknanopolymer was used to package the new formulation of Curcumin and Tamoxifen. Anti-cancer efficacy of the obtained compound was evaluated in Tamoxifen-sensitive (TS). MCF-7, Tamoxifen-resistant (TR) MCF-7 cancer cells and Fibroblast cells. MTT assay was used to evaluate anti-proliferation and toxicity. Flow cytometry and Annexin-V-FLUOS were used to assay anti-proliferation and induction of apoptosis respectively. Our results indicate that the obtained nano-compound is less toxic to normal cells compared to Tamoxifen alone, and has higher anti-proliferation and pro-apoptotic activity on TS-MCF-7 and TR-MCF-7. The nanopolymer reduces the Tamoxifen toxicity in normal cells and counters the developed resistance to the drug in cancer cells.


Laparoscopic management of a newborn with a right Amyand's hernia and a left incarcerated inguinal hernia.
A one month old boy presented with left incarcerated inguinal hernia. After unsuccessful manual reduction, we decided to perform laparoscopic herniorrhaphy. Laparoscopic examination showed a left hernia with intestinal loops that entered into the internal inguinal ring, on the right side there was an unknown patency of the peritoneal vaginal duct with the appendix completely incarcerated within the sac. On the left side, the loops were reduced with a combined technique of external manual pressure and internal pulling by forceps; the bowel was inspected, and the hernia was repaired. On the right side, the appendix was strongly adherent with the peritoneal vaginal duct, and the reduction was not possible. The appendix was dissected from the sac using a 3-mm monopolar hook and than reduced into the abdomen, then right herniorrhaphy was performed. Two days after surgery, the baby had fever and abdominal distension. He was re-operated through mini-Pfannenstiel incision and an ischemic appendix was identified and removed. Postoperative period was uneventful. In our case, laparoscopy allowed for simultaneous reduction under direct visual control, inspection of the incarcerated organ, definitive repair and incidentally discovery and treatment of the contralateral incarcerated Amyand's hernia. In case of incarcerated appendix, appendectomy is preferable during the same procedure to reduce the incidence of postoperative complications.


Ocular safety of sildenafil citrate when administered chronically for pulmonary arterial hypertension: results from phase III, randomised, double masked, placebo controlled trial and open label extension.
To assess the ocular effects and safety profile of chronic sildenafil oral dosing in patients with pulmonary arterial hypertension. 12 week, double masked, randomised, placebo controlled, phase III trial with open label extension. 53 institutions worldwide. 277 adults with idiopathic pulmonary arterial hypertension or pulmonary arterial hypertension associated with connective tissue disease or after congenital heart disease repair (mean pulmonary artery pressure ≥25 mm Hg; pulmonary capillary wedge pressure ≤15 mm Hg at rest). During the double masked study, oral sildenafil 20 mg, 40 mg, or 80 mg or placebo (1:1:1:1) three times daily for 12 weeks was added to baseline drug treatment. In the extension study, the placebo, 20 mg and 40 mg groups received 40 mg three times daily titrated to 80 mg three times daily at week 6. After unmasking, the dose was titrated according to clinical need. Ocular safety (ocular examinations, visual function tests, participants' reports of adverse events, and visual disturbance questionnaire completed by investigators) by treatment group at 12 weeks, 24 weeks, 18 months, and yearly. Findings of the objective assessments-that is, intraocular pressure and visual function tests (visual acuity, colour vision, and visual field)-were similar across groups (20 mg, n=69; 40 mg, n=67; 80 mg, n=71; placebo, n=70). No clinically significant changes occurred between baseline and 12 weeks, except for an efficacy signal in contrast sensitivity for the sildenafil 40 mg three times daily group. In right eyes, changes in intraocular pressure from baseline to week 12 ranged from a mean of -0.5 (95% confidence interval -1.3 to 0.2) mm Hg with placebo, -0.2 (-0.9 to 0.5) mm Hg with sildenafil 40 mg, and -0.1 (-0.7 to 0.5) mm Hg with 80 mg to 0.3 (-0.4 to 0.9) mm Hg with sildenafil 20 mg (the approved dose for pulmonary arterial hypertension). Mean changes from baseline to week 12 in contrast sensitivity in right eyes were -0.02 (SD 0.12) in the sildenafil 20 mg three times daily group compared with -0.05 (0.18) in the placebo group (P=0.044). Percentages of participants with deterioration in visual acuity (Snellen) from baseline to week 12 ranged from 10% (n=7) in the placebo group to 3% (n=2) in the sildenafil 20 mg three times daily group; the same percentages had visual field changes from normal to abnormal during the period in these two groups. The investigators did not deem any findings on colour vision assessment to be clinically significant. Findings of the objective assessments in the 40 mg and 80 mg three times daily sildenafil treatment groups and in left eyes were not substantially different, nor were any measures different throughout the open label extension compared with week 12. However, objective data were limited after month 18, as most participants had missing data or visual parameters were no longer collected by investigators. Incidence of ocular adverse events reported on the case report forms and assessed by the investigator was low with all doses, but a modest, dose related incidence of chromatopsia, cyanopsia, photophobia, and visual disturbance was reported with 80 mg three times daily consistent with the indicated dosing for erectile dysfunction. Retinal haemorrhages, captured on funduscopy, occurred in 2% (4/207) of sildenafil treated participants and none in the placebo group during the double masked study and in 4% (10/259) during the open label extension. Sildenafil dosing up to 80 mg three times daily is safe and well tolerated from an ocular perspective in patients with pulmonary arterial hypertension. Daily chronic dosing in this patient population was not associated with visual change and had no detrimental effect on best corrected visual acuity, contrast sensitivity, colour vision, or visual field, or on slit lamp examinations, funduscopy, or intraocular pressure during the duration of this study. Clinical trials NCT00644605 and NCT00159887.


CD4(+) and CD8(+) regulatory T cells in chronic rhinosinusitis mucosa.
Chronic rhinosinusitis (CRS) mucosal inflammation is characterized by an accumulation of effector-memory T cells, but their immune regulatory potential has not been adequately examined. Coexpression of transcription factor, forkhead box P3 (Foxp3), and interleukin-2 receptor, CD25, in CD4(+) and CD8(+) T cells is linked with regulatory function in humans. The aim of this study was to investigate the regulatory T cell (Treg) phenotype of CD4(+) (CD4Treg) and CD8(+) (CD8Treg) T cells in peripheral blood (PB) and sinus mucosa of CRS patients. Prospective study was performed involving 32 CRS with nasal polyp (CRSwNP), 14 CRS without nasal polyp (CRSsNP), and 8 control patients. Sinus and PB T lymphocytes were stained with CD3, CD4, CD8, CD25, and Foxp3 and analyzed using flow cytometry. Relevant clinical characteristics, sinus bacterial culture results, and eosinophilic mucus were examined. Sinus mucosa had a higher percentage of CD4Treg (CD3(+)CD4(+)CD25(+)Foxp3(+)) population compared with PB in all patients. The percentage of PB CD4Treg and CD8Treg (CD3(+)CD8(+)CD25(+)Foxp3(+)) was not significantly different between the study groups. CRS mucosal tissue had a higher percentage of CD4Treg and activated T-helper cells than controls. There was no significant difference in PB and mucosal CD4Treg populations in CRS patients based on the presence of allergy, sinus culture results, or eosinophilic mucus. In controls, increased mucosal CD4Treg correlated with coexisting allergy. Although overall CD4Treg numbers were higher, the regulatory potential of activated CD4(+) T cells (CD4Treg/activated T-helper cell ratio) was significantly lower in CRS mucosa compared with controls. The CD8Treg subset was also significantly reduced in CRSwNP mucosa compared with controls. A higher percentage of CD4Treg and activated T-helper cells in CRS mucosa suggests increased inflammation in CRS, independent of the presence of allergy, microbial culture results, or eosinophilic mucus. However, the decreased ratio of CD4Treg versus activated T-helper cells in CRS and reduced CD8Treg population in CRSwNPs indicates an inflammatory bias and the inability to control mucosal disease.


Ensemble place codes in hippocampus: CA1, CA3, and dentate gyrus place cells have multiple place fields in large environments.
Previously we reported that the hippocampus place code must be an ensemble code because place cells in the CA1 region of hippocampus have multiple place fields in a more natural, larger-than-standard enclosure with stairs that permitted movements in 3-D. Here, we further investigated the nature of hippocampal place codes by characterizing the spatial firing properties of place cells in the CA1, CA3, and dentate gyrus (DG) hippocampal subdivisions as rats foraged in a standard 76-cm cylinder as well as a larger-than-standard box (1.8 m×1.4 m) that did not have stairs or any internal structure to permit movements in 3-D. The rats were trained to forage continuously for 1 hour using computer-controlled food delivery. We confirmed that most place cells have single place fields in the standard cylinder and that the positional firing pattern remapped between the cylinder and the large enclosure. Importantly, place cells in the CA1, CA3 and DG areas all characteristically had multiple place fields that were irregularly spaced, as we had reported previously for CA1. We conclude that multiple place fields are a fundamental characteristic of hippocampal place cells that simplifies to a single field in sufficiently small spaces. An ensemble place code is compatible with these observations, which contradict any dedicated coding scheme.


Enhanced Raman scattering from nanoparticle-decorated nanocone substrates: a practical approach to harness in-plane excitation.
We investigate surface-enhanced Raman scattering (SERS) from gold-coated silicon-germanium nanocone substrates that are decorated with 30-nm spherical gold nanoparticles (AuNPs). Finite-element simulations suggest that individual nanocones generate stronger electromagnetic enhancement with axial polarization (i.e., polarization parallel to the vertical axis of the nanocones) than with transverse polarization (i.e., polarization in the plane of the nanocone substrate), whereas the excitation in a typical Raman microscope is mainly polarized in the transverse plane. We introduce a practical approach to improve the SERS performance of the substrate by filling the valleys between nanocones with AuNPs. Simulations reveal an enhanced electric field at the nanoscale junctions formed between AuNPs and nanocones, and we explain this lateral coupling with a hybridization model for a particle-film system. We further experimentally verify the added enhancement by measuring SERS from trans-1,2-bi-(4-pyridyl) ethylene molecules absorbed onto the substrates. We report over one order-of-magnitude increase in SERS activities with the AuNP decoration (compared to the nanocone substrate without AuNPs) and achieve a spatially averaged enhancement factor of 1.78 × 10(8) at 785-nm excitation. Understanding and implementing the enhancing mechanism of structured metallic surfaces decorated with plasmonic nanoparticles open possibilities to substantially improve the SERS performance of the existing process-engineered substrates.


Juggling work and breastfeeding: effects of maternity leave and occupational characteristics.
Juggling breastfeeding and paid work can challenge breastfeeding success. We examined the relationship between breastfeeding and maternity leave before and after delivery among working mothers in Southern California. California is 1 of only 5 states in the United States providing paid pregnancy leave that can be extended for infant bonding. Drawing from a case-control study of preterm birth and low birth weight, 770 full-time working mothers were compared on whether they established breastfeeding in the first month. For those who established breastfeeding, we examined duration. Eligible women participated in California's Prenatal Screening Program; delivered live births between July 2002 and December 2003; were > or =18 years old; had a singleton birth without congenital anomalies; and had a US mailing address. We assessed whether maternity leave and other occupational characteristics predicted breastfeeding cessation and used multivariate regression models weighted for probability of sampling to calculate odds ratios for breastfeeding establishment and hazards ratios for breastfeeding cessation. A maternity leave of < or =6 weeks or 6 to 12 weeks after delivery was associated, respectively, with a fourfold and twofold higher odds of failure to establish breastfeeding and an increased probability of cessation after successful establishment, relative to women not returning to work, after adjusting for covariates. The impact of short postpartum leave on breastfeeding cessation was stronger among nonmanagers, women with inflexible jobs, and with high psychosocial distress. Antenatal leave in the last month of pregnancy was not associated with breastfeeding establishment or duration. Postpartum maternity leave may have a positive effect on breastfeeding among full-time workers, particularly those who hold nonmanagerial positions, lack job flexibility, or experience psychosocial distress. Pediatricians should encourage patients to take maternity leave and advocate for extending paid postpartum leave and flexibility in working conditions for breastfeeding women.


Morphoproteomics and Biomedical Analytics Identify the c-Myc, EZH2, Sirt1 and CXCR4 Pathways as Potential Blocks to Differentiation Leading to Proliferation and Metastatic Potential in Sinonasal Undifferentiated Carcinoma (SNUC) and Provide Therapeutic Options: A Case Study.
Sinonasal undifferentiated carcinoma (SNUC) is a highly malignant tumor in the nasal cavity or paranasal sinuses. Morphoproteomics has defined its biology to some degree, allowing the identification of targeted therapeutic options with clinical efficacy [1]. This study's objective was to identify putative SNUC pathways that are known to pose a block in differentiation both in early embryogenesis and in tumorigenesis or that might promote metastasis and recurrent disease. Morphoproteomic analysis of SNUC from a case study of this patient included immunohistochemical probes to detect c-Myc, EZH2, Sirt1 and CXCR4 protein analytes. Biomedical analytics schematically showed the interactions of these analytes with the morphoproteomic findings and illustrated targeted therapeutic options. Representative sections of this patient's tumor displayed plasmalemmal expression for CXCR4 and nuclear immunopositivity for c-Myc, EZH2, and Sirt1. This coincided with their block in differentiation and their proliferative state with progression into the mitotic phase. Biomedical analytics integrated the morphoproteomic findings with the undifferentiated and proliferative state of SNUC and depicted pharmacogenomic and other related factors that target the c-Myc, EZH2, Sirt1 and CXCR4 pathways. Morphoproteomics and biomedical analytics have identified c-Myc, EZH2, Sirt1 and CXCR4 pathways that collectively could contribute to the block in differentiation and increase the propensity for recurrence and metastasis in SNUC. This suggests that combinatorial therapies modulating these pathways could be used in a maintenance mode to minimize the risk of recurrent disease.


Bridging neurocognitive aging and disease modification: targeting functional mechanisms of memory impairment.
Risk for Alzheimer's disease escalates dramatically with increasing age in the later decades of life. It is widely recognized that a preclinical condition in which memory loss is greater than would be expected for a person's age, referred to as amnestic mild cognitive impairment, may offer the best opportunity for intervention to treat symptoms and modify disease progression. Here we discuss a basis for age-related memory impairment, first discovered in animal models and recently isolated in the medial temporal lobe system of man, that offers a novel entry point for restoring memory function with the possible benefit in slowing progression to Alzheimer's disease.


Volumetric and densitometric study in the cerebral cortex and septum of a lizard (Lacerta galloti) using the Timm method.
The Timm sulfide-silver method for the histochemical detection of heavy metals has been applied to the study of cortical and septal regions in the telencephalon of the lizard Lacerta galloti. Reacting areas are located in the anterodorsal septum, in the inner plexiform layer of the medial cortex, and in the plexiform layers of the dorsomedial and dorsal cortices. A very specific kind of axonal ending is responsible for this reactivity. Volumetric and densitometric measures have been performed in each reacting area. Usefulness of both measures as indicators of the number of labeled axonal endings projecting on a zone is discussed. Resemblance between these axonal endings and hippocampal mossy fiber boutons in mammals is suggested.


HobPre: accurate prediction of human oral bioavailability for small molecules.
Human oral bioavailability (HOB) is a key factor in determining the fate of new drugs in clinical trials. HOB is conventionally measured using expensive and time-consuming experimental tests. The use of computational models to evaluate HOB before the synthesis of new drugs will be beneficial to the drug development process. In this study, a total of 1588 drug molecules with HOB data were collected from the literature for the development of a classifying model that uses the consensus predictions of five random forest models. The consensus model shows excellent prediction accuracies on two independent test sets with two cutoffs of 20% and 50% for classification of molecules. The analysis of the importance of the input variables allowed the identification of the main molecular descriptors that affect the HOB class value. The model is available as a web server at www.icdrug.com/ICDrug/ADMET for quick assessment of oral bioavailability for small molecules. The results from this study provide an accurate and easy-to-use tool for screening of drug candidates based on HOB, which may be used to reduce the risk of failure in late stage of drug development.


[Interventional cardiology in pediatrics].
Since the first description of successful pulmonary balloon valvuloplasty in 1982, the field of pediatric interventional cardiology has seen a dramatic development Nowadays, interventional techniques are a standard therapy for a wide range of indications in congenital heart disease: balloon pulmonary and aortic valvuloplasty are therapies of choice as well as the occlusion of the persistent ductus arteriosus by double umbrellas or coils. Dilatation of a recoarctation after initial surgical therapy is safe and successful whereas there is an ongoing debate whether native coarctation should be managed surgically or by catheter intervention. Other common indications for catheter interventions are dilatation and stenting of native or postoperative vessel stenosis and occlusion of abnormal intrathoracic vessels with coils. Other indications such as closure of atrial septal defects are about to become clinical routine, whereas interventional closure of ventricular septal defects must still be considered an experimental approach. Moreover in complex congenital heart disease there are a number of other indications for very specific interventional techniques. Interventional therapies should be seen either as adjuvant to surgical management in some cases or as an alternative approach to avoid surgery in others. It is important that the indications for surgical or interventional therapy result from a discussion involving the pediatric cardiologist and the pediatric cardiac surgeon.


Cerebral arterial thrombosis in a child with nephrotic syndrome.
Nephrotic syndrome (NS) in childhood may be associated with thromboembolic complications, mainly in venous origin. However, arterial thrombosis may also be seen as a very rare and life-threatening complication. Herein, we described a case of steroid-resistant NS who did not respond to full-dose steroid treatment for 8 weeks and was complicated by neurological findings. The renal biopsy was consistent with focal segmental glomerulosclerosis. His cerebral magnetic resonance angiography showed the sudden termination of M3 branch of the left middle cerebral artery which corresponded with subacute infarction in the left frontoparietotemporal area. Thrombosis panel yielded the results of hyperhomocysteinemia (46.1 μmol/L, range: 5-15 μmol/L) and heterozygous methylene tetrahydrofolate reductase mutation (C677T, A1298C). After that, the patient was given medical therapy including anticoagulant treatment. Improvement in the neurological outcome was determined on the 1st month of follow-up examinations.


Temporal changes in renal glomerular function associated with the use of Tenofovir Disoproxil Fumarate in HIV-infected Nigerians.
Reports of renal dysfunction in Tenofovir Disoproxil Fumarate (TDF)-treated HIV-1 infected patients have raised concerns about potential nephrotoxicity. To compare the effects on renal function of TDF-containing highly active anti-retroviral therapy (HAART) with a non-TDF-containing HAART. This was an observational study.Clinical and laboratory data of 186 HIV-1 infected adult Nigerians on first-line HAART for at least 48 weeks were reviewed. Eighty-four patients whose nucleos(t)ide reverse transcriptase inhibitor (NRTI) backbone included TDF were compared to 102 patients on other NRTI backbones. Creatinine clearance (CLcr) was estimated using the Cockcroft-Gault equation. Changes in serum creatinine and CLcr from the baseline for each patient were compared between the TDF-treated and the TDF-free patients. We also assessed the associations of other variables with change in CLcr... Baseline median serum creatinine (mmol/L) was 77 and 84 in the TDF-treated and TDF-free groups, respectively (p=0.59). Baseline median CLcr (mls/min) was 83 in the TDF-treated patients vs 78 in the TDF-free group. At 48 weeks, serum creatinine increased by 18.1% and 1.2% in the TDF-treated and TDF-free arms, respectively. There was a decrease of 4.8% in GFR in the TDF arm compared to a gain 5.1% in the TDF-free arm. Tenofovir Disoproxil Fumarate-containing HAART is associated with a slight decline in the medium term in CLcr compared with HAART regimens containing alternative Nucleosid(t) Reverse Transcriptase Inhibitors.


Changes in monoamine metabolites concentrations in rat cerebrospinal fluid after acute and long-term administration of a selective serotonin reuptake inhibitor, trazodone.
In order to clarify the mechanism of the antidepressive effects of trazodone, a selective serotonin reuptake inhibitor, we investigated the dynamics of monoamine metabolites in cerebrospinal fluid (CSF) of free-moving conscious rats by acute and long-term treatment with trazodone. When 100 mg kg-1 p.o. of trazodone were administered, a significant increase of 3-methoxy-4-hydroxyphenylglycol (MHPG) concentration was soon observed in the light period of the light/dark cycle, and a significant decrease of dihydroxy phenyl acetic acid (DOPAC) concentration was observed during the 2 days after administration of trazodone; in contrast, the homovanilic acid (HVA) level was increased. However, we detected no significant changes in the 5-hydroxy indole-3-acetic acid (5-HIAA) concentration during the 3 days. In the case of long-term treatment with 50 mg kg-1, p.o. of trazodone, the levels of MHPG, DOPAC and HVA exhibited no difference when compared with values obtained during saline treatment in either the light or dark period, whereas the levels of 5-HIAA showed a significant increase during the light period. These findings suggest that a long-term treatment with trazodone enhances the serotonergic neurons.


Pattern formation in the vertebrate neural tube: a sonic hedgehog morphogen-regulated transcriptional network.
Neuronal subtype specification in the vertebrate neural tube is one of the best-studied examples of embryonic pattern formation. Distinct neuronal subtypes are generated in a precise spatial order from progenitor cells according to their location along the anterior-posterior and dorsal-ventral axes. Underpinning this organization is a complex network of multiple extrinsic and intrinsic factors. This review focuses on the molecular mechanisms and general strategies at play in ventral regions of the forming spinal cord, where sonic hedgehog-based morphogen signaling is a key determinant. We discuss recent advances in our understanding of these events and highlight unresolved questions.


Negative feedback may suppress variation to improve collective foraging performance.
Social insect colonies use negative as well as positive feedback signals to regulate foraging behaviour. In ants and bees individual foragers have been observed to use negative pheromones or mechano-auditory signals to indicate that forage sources are not ideal, for example being unrewarded, crowded, or dangerous. Here we propose an additional function for negative feedback signals during foraging, variance reduction. We show that while on average populations will converge to desired distributions over forage patches both with and without negative feedback signals, in small populations negative feedback reduces variation around the target distribution compared to the use of positive feedback alone. Our results are independent of the nature of the target distribution, providing it can be achieved by foragers collecting only local information. Since robustness is a key aim for biological systems, and deviation from target foraging distributions may be costly, we argue that this could be a further important and hitherto overlooked reason that negative feedback signals are used by foraging social insects.


The changing face of chronic bronchitis with airways obstruction.
A total of 178 patients with obstructive airways disease were studied between 1966 and 1972, a period which followed a substantial fall in atmospheric pollution in Sheffield. Compared with patients from an earlier study of the same age and smoking habits they had less productive cough, fewer winter illnesses, less severe breathlessness, and only one-third of the rate of decline of forced expiratory volume (FEV). The analysis of the change in the annual loss of FEV was complicated. There were virtually no sudden steep falls of respiratory function, and patients with very low values of FEV survived for long periods. The predominant cause of death was cor pulmonale.Chronic hypoxia may be more of a problem in the next few years than bronchial infection. I believe that the clean air policy has effected an improvement and that the nature of chronic bronchitis with severe obstructive airways disease is changing, but further studies will be needed to confirm this.


Innate defense against influenza A virus: activity of human neutrophil defensins and interactions of defensins with surfactant protein D.
Surfactant protein D (SP-D) plays important roles in innate host defense against influenza A virus (IAV) infection, in part by modifying interactions with neutrophils. Human neutrophil defensins (HNPs) inhibit infectivity of enveloped viruses, including IAV. Our goal in this study was to characterize antiviral interactions between SP-D and HNPs. Recombinant and/or natural forms of SP-D and related collectins and HNPs were tested for antiviral activity against two different strains of IAV. HNPs 1 and 2 did not inhibit viral hemagglutination activity, but they interfered with the hemagglutination-inhibiting activity of SP-D. HNPs had significant viral neutralizing activity against divergent IAV strains. However, the HNPs generally had competitive effects when combined with SP-D in assays using an SP-D-sensitive IAV strain. In contrast, cooperative antiviral effects were noted in some instances when relatively SP-D-resistant strains were treated with SP-D and HNPs. HNPs were found to bind to the neck and/or carbohydrate recognition domain of SP-D. This binding was specific because no, or minimal, binding to other collectins was found. HNPs precipitated SP-D from bronchoalveolar lavage fluid and reduced the antiviral activity of bronchoalveolar lavage fluid. HNP-1 and -2 differed somewhat in their independent antiviral activity and their binding to SP-D. These results are relevant to the early phase of host defense against IAV, and suggest a complex interplay between SP-D and HNPs at sites of active inflammation.


Hedgehog signaling is necessary and sufficient to mediate craniofacial plasticity in teleosts.
Phenotypic plasticity, the ability of a single genotype to produce multiple phenotypes under different environmental conditions, is critical for the origins and maintenance of biodiversity; however, the genetic mechanisms underlying plasticity as well as how variation in those mechanisms can drive evolutionary change remain poorly understood. Here, we examine the cichlid feeding apparatus, an icon of both prodigious evolutionary divergence and adaptive phenotypic plasticity. We first provide a tissue-level mechanism for plasticity in craniofacial shape by measuring rates of bone deposition within functionally salient elements of the feeding apparatus in fishes forced to employ alternate foraging modes. We show that levels and patterns of phenotypic plasticity are distinct among closely related cichlid species, underscoring the evolutionary potential of this trait. Next, we demonstrate that hedgehog (Hh) signaling, which has been implicated in the evolutionary divergence of cichlid feeding architecture, is associated with environmentally induced rates of bone deposition. Finally, to demonstrate that Hh levels are the cause of the plastic response and not simply the consequence of producing more bone, we use transgenic zebrafish in which Hh levels could be experimentally manipulated under different foraging conditions. Notably, we find that the ability to modulate bone deposition rates in different environments is dampened when Hh levels are reduced, whereas the sensitivity of bone deposition to different mechanical demands increases with elevated Hh levels. These data advance a mechanistic understanding of phenotypic plasticity in the teleost feeding apparatus and in doing so contribute key insights into the origins of adaptive morphological radiations.


Photosynthetic benefits of ultraviolet-A to Pimelea ligustrina, a woody shrub of sub-alpine Australia.
The definition of photosynthetically active radiation (Q) as the visible waveband (λ 400-700 nm) is a core assumption of much of modern plant biology and global models of carbon and water fluxes. On the other hand, much research has focused on potential mutation and damage to leaves caused by ultraviolet (UV) radiation (280-400 nm), and anatomical and physiological adaptations that help avoid such damage. Even so, plant responses to UV-A are poorly described and, until now, photosynthetic utilization of UV-A has not been elucidated under full light conditions in the field. We found that the UV-A content of sunlight increased photosynthetic rates in situ by 12% in Pimelea ligustrina Labill., a common and indigenous woody shrub of alpine ecosystems of the Southern Hemisphere. Compared to companion shrubs, UV-A-induced photosynthesis in P. ligustrina resulted from reduced physical and chemical capacities to screen UV-A at the leaf surface (illustrated by a lack of cuticle and reduced phenol index) and the resulting ability of UV-A to excite chlorophyll (Chl) a directly, and via energy provided by the carotenoid lutein. A screening of 55 additional sub-alpine species showed that 47% of the plant taxa also display Chl a fluorescence under UV-A. If Chl a fluorescence indicates potential for photosynthetic gain, continued exclusion of UV-A from definitions of Q in this ecosystem could result in underestimates of measured and modeled rates of photosynthesis and miscalculation of potential for carbon sequestration. We suggest that carbon gain for alpine environs across the globe could be similarly underestimated given that UV-A radiation increases with altitude and that the frequently dominant herb and grass life-forms often transmit UV-A through the epidermis.


Residents' asthma communication skills in announced versus unannounced standardized patient exercises.
The aim of this study was to test the hypothesis that pediatric residents would display similar levels of asthma interpersonal and communication skills in announced versus unannounced adolescent standardized patient (SP) encounters. We conducted a prospective repeat measures experimental study at a pediatric residency program at an inner-city children's hospital. A cohort of residents (N = 18) was subjected at random to 3 SP exercises: announced and being directly observed by faculty, announced and not observed by faculty, and unannounced and unobserved. Six adolescent SPs were trained to complete checklists that included items like asthma daytime and nighttime symptoms, exercise-induced symptoms, triggers, and asthma education. For the unannounced exercises, SPs were inserted into residents' regularly scheduled clinics. Standardized patients rated residents immediately following each exercise. Residents were rated by faculty following the observed encounter. Faculty rating validated SP ratings on the observed encounter. Differences in proportions of categorical variables were tested by chi-square analyses. Fifty-four resident-SP encounters were analyzed. Residents consistently displayed significantly lower levels of desired behaviors in interpersonal and communication skills in the unannounced SP encounters on 10 of 14 checklist items. For example, residents asked about exercise-induced symptoms 90% of the time in announced/observed encounters versus 95% in announced/unobserved encounters versus 72% in unannounced/unobserved encounters (P = .001). There were no significant differences in residents' behaviors in the announced SP exercises (whether observed or unobserved). In this study, residents demonstrated lower levels of asthma communication skills during unannounced SP exercises. By using unannounced SPs, we were able to assess residents' interpersonal and communication skills in real clinical settings.


Independent and overlapping transcriptional activation during liver development and regeneration in mice.
Liver development and regeneration share the requirement for simultaneous proliferation and acquisition of highly specialized cellular functions. However, little is known about molecules with regulatory roles in both processes. We hypothesized that transcriptional reprogramming induced by regeneration recapitulates that of developing liver. To address this hypothesis, we determined global hepatic gene expression at embryonic day 14.5, postnatal day 14, and 6 to 24 hours following partial hepatectomy using microarrays containing 8,635 cDNAs. Analysis of genes overexpressed during these conditions revealed 3 unique expression patterns. The first was predominantly signature gene clusters specific for each growth phase. Major groups were hematopoiesis-related genes in embryonic livers, metabolic genes during postnatal liver development, and growth/inflammation and metabolic genes during regeneration. The second pattern consisted of dual overexpression during regeneration and at least one phase of development. Consistent with potential regulatory roles in liver growth, most of these transcripts control cell-cell contact, membrane trafficking, cell growth, metabolism, and inflammatory response. The third pattern, revealed by surveying their expression across 76 hepatic and extra-hepatic tissues, uncovered a restricted temporospatial pattern of liver overexpression for CD14, orosomucoid 1, hepcidin, Spi 2.1, Ith3, and Tim-44. In conclusion, these results provide a basis for the identification of gene and gene groups that play critical roles at different phases of liver development and regeneration, and underscore the importance of maintaining metabolic demands during organ growth.


Sequence elements upstream of the 3' cleavage site confer substrate strength to the adenovirus L1 and L3 polyadenylation sites.
The adenovirus major late transcription unit is a well-characterized transcription unit which relies heavily on alternative pre-mRNA processing to generate distinct populations of mRNA during the early and late stages of viral infection. In the early stage of infection, two major late transcription unit mRNA transcripts are generated through use of the first (L1) of five available poly(A) sites (L1 through L5). This contrasts with the late stage of infection when as many as 45 distinct mRNAs are generated, with each of the five poly(A) sites being used. In previous work characterizing elements involved in alternative poly(A) site use, we showed that the L1 poly(A) site is processed less efficiently than the L3 poly(A) site both in vitro and in vivo. Because of the dramatic difference in processing efficiency and the role processing efficiency plays in production of steady-state levels of mRNA, we have identified the sequence elements that account for the differences in L1 and L3 poly(A) site processing efficiency. We have found that the element most likely to be responsible for poly(A) site strength, the GU/U-rich downstream element, plays a minor role in the different processing efficiencies observed for the L1 and L3 poly(A) sites. The sequence element most responsible for inefficient processing of the L1 poly(A) site includes the L1 AAUAAA consensus sequence and those sequences which immediately surround the consensus hexanucleotide. This region of the L1 poly(A) site contributes to an inability to form a stable processing complex with the downstream GU/U-rich element. In contrast to the L1 element, the L3 poly(A) site has a consensus hexanucleotide and surrounding sequences which can form a stable processing complex in cooperation with the downstream GU/U-rich element. The L3 poly(A) site is also aided by the presence of sequences upstream of the hexanucleotide which facilitate processing efficiency. The sequence UUCUUUUU, present in the L3 upstream region, is shown to enhance processing efficiency as well as stable complex formation (shown by increased binding of the 64-kDa cleavage stimulatory factor subunit) and acts as a binding site for heterogeneous nuclear ribonucleoprotein C proteins.


Advanced Research Institute (ARI): Supporting the Geriatric Mental Health Research Pipeline.
The Advanced Research Institute (ARI) in Mental Health and Aging is a NIMH-funded mentoring network to help transition early-career faculty to independent investigators and scientific leaders. Since 2004, ARI has enrolled 184 Scholars from 61 institutions across 34 states. We describe the ARI components and assess the impact and outcomes of ARI on research careers of participants. Outcomes of ARI graduates (n = 165) came from NIH Reporter, brief surveys, and CVs: 87.3% remained active researchers, 83.6% performed scientific service, and 80.6% obtained federal grants. A population-based analysis examined NIMH mentored K awardees initially funded from 2002-2018 (n = 1160): in this group, 77.1% (47/61) of ARI participants versus 49.5% (544/1099) of nonparticipants obtained an R01. Controlling for time, ARI participants were 3.2 times more likely to achieve R01 funding than nonparticipants. Given the struggle to reduce attrition from the research career pipeline, the effectiveness of ARI model could be relevant to other fields.


Loss of T-bet confers survival advantage to influenza-bacterial superinfection.
The transcription factor, T-bet, regulates type 1 inflammatory responses against a range of infections. Here, we demonstrate a previously unaddressed role of T-bet, to influenza virus and bacterial superinfection. Interestingly, we found that T-bet deficiency did not adversely affect the efficacy of viral clearance or recovery compared to wild-type hosts. Instead, increased infiltration of neutrophils and production of Th17 cytokines (IL-17 and IL-22), in lungs of influenza virus-infected T-bet-/- mice, were correlated with survival advantage against subsequent infection by Streptococcus pneumoniae Neutralization of IL-17, but not IL-22, in T-bet-/- mice increased pulmonary bacterial load, concomitant with decreased neutrophil infiltration and reduced survival of T-bet-/- mice. IL-17 production by CD8+, CD4+ and γδ T cell types was identified to contribute to this protection against bacterial superinfection. We further showed that neutrophil depletion in T-bet-/- lungs increased pulmonary bacterial burden. These results thus indicate that despite the loss of T-bet, immune defences required for influenza viral clearance are fully functional, which in turn enhances protective type 17 immune responses against lethal bacterial superinfections.


Analyzing metabolomics data for association with genotypes using two-component Gaussian mixture distributions.
Standard approaches to evaluate the impact of single nucleotide polymorphisms (SNP) on quantitative phenotypes use linear models. However, these normal-based approaches may not optimally model phenotypes which are better represented by Gaussian mixture distributions (e.g., some metabolomics data). We develop a likelihood ratio test on the mixing proportions of two-component Gaussian mixture distributions and consider more restrictive models to increase power in light of a priori biological knowledge. Data were simulated to validate the improved power of the likelihood ratio test and the restricted likelihood ratio test over a linear model and a log transformed linear model. Then, using real data from the Framingham Heart Study, we analyzed 20,315 SNPs on chromosome 11, demonstrating that the proposed likelihood ratio test identifies SNPs well known to participate in the desaturation of certain fatty acids. Our study both validates the approach of increasing power by using the likelihood ratio test that leverages Gaussian mixture models, and creates a model with improved sensitivity and interpretability.


Distribution of cholecystokinin-8-like immunoreactivity in the frog brain and spinal cord.
The distribution of cholecystokinin-8 (CCK8) containing structures in the central nervous system of the frog (Rana esculenta) was investigated with the aid of the peroxidase immunohistochemical method. In the forebrain, immunoreactive perikarya were found in the main olfactory bulb, septum, amygdala, and ventromedial thalamic nuclei. Networks of immunoreactive varicose fibers were observed in the septum, the basal area, and the infundibulum, and fiber bundles were found in the marginal and axial optic tracts. In the midbrain, immunoreactive cell bodies were seen in the mesencephalic reticular nucleus, and networks of fibers were observed in the optic tectum and the tegmentum. In the hindbrain, immunoreactive neurons were found in the nucleus of the solitary tract, and varicose fibers were observed in the superior vestibular nucleus and the subventricular area. The gray matter of the spinal cord was interlaced by immunoreactive fibers, and a few intensely stained perikarya were seen in the base of the dorsal horn. These findings indicate that CCK8 is widely distributed in the central nervous system of the frog, like in mammals. The CCK8 immunoreactivity is particularly high in the limbic structures and some diencephalic and mesencephalic areas (infundibulum, optic tectum), whereas the pallium is devoid of this peptide in the frog.


Vgsc-interacting proteins are genetically associated with pyrethroid resistance in Aedes aegypti.
Association mapping of factors that condition pyrethroid resistance in Aedes aegypti has consistently identified genes in multiple functional groups. Toward better understanding of the mechanisms involved, we examined high throughput sequencing data (HTS) from two Aedes aegypti aegypti collections from Merida, Yucatan, Mexico treated with either permethrin or deltamethrin. Exome capture enrichment for coding regions and the AaegL5 annotation were used to identify genes statistically associated with resistance. The frequencies of single nucleotide polymorphisms (SNPs) were compared between resistant and susceptible mosquito pools using a contingency χ2 analysis. The -log10(χ2 p value) was calculated at each SNP site, with a weighted average determined from all sites in each gene. Genes with -log10(χ2 p value) ≥ 4.0 and present among all 3 treatment groups were subjected to gene set enrichment analysis (GSEA). We found that several functional groups were enriched compared to all coding genes. These categories were transport, signal transduction and metabolism, in order from highest to lowest statistical significance. Strikingly, 21 genes with demonstrated association to synaptic function were identified. In the high association group (n = 1,053 genes), several genes were identified that also genetically or physically interact with the voltage-gated sodium channel (VGSC). These genes were eg., CHARLATAN (CHL), a transcriptional regulator, several ankyrin-domain proteins, PUMILIO (PUM), a translational repressor, and NEDD4 (E3 ubiquitin-protein ligase). There were 13 genes that ranked among the top 10%: these included VGSC; CINGULIN, a predicted neuronal gap junction protein, and the aedine ortholog of NERVY (NVY), a transcriptional regulator. Silencing of CHL and NVY followed by standard permethrin bottle bioassays validated their association with permethrin resistance. Importantly, VGSC levels were also reduced about 50% in chl- or nvy-dsRNA treated mosquitoes. These results are consistent with the contribution of a variety of neuronal pathways to pyrethroid resistance in Ae. aegypti.