Conotruncal malformations and absent thymus due to a deleterious NKX2-6 mutation.
Truncus arteriosus (TA) accounts for ~1% of congenital heart defects. The aetiology of isolated TA is largely unknown but when occurring as part of a syndrome, it is mostly associated with chromosome 22q11 deletion. Vice versa, the most common congenital heart defects associated with chromosome 22q11 deletion are conotruncal malformations. In this study we investigated the cause of multiple conotruncal malformations accompanied by athymia in a consanguineous family. Whole exome analysis revealed a homozygous deleterious mutation in the NKX2-6 gene. NKX2-6 encodes a homeobox-containing protein which is expressed in mouse embryo at E8.0-E9.5 at the caudal pharyngeal arches and the outflow tract. A single missense mutation was previously implicated in the aetiology of familial isolated TA; however, null mice are entirely normal. The clear phenotype associated with a homozygous deleterious mutation in the present report, falls well within the spectrum of the cardiac defects seen in DiGeorge syndrome, is in agreement with NKX2-6 downstream location in the TBX1 signalling pathway and confirms NKX2-6 role in human cardiogenesis.


Introduction to antibody engineering and phage display.
The vertebrate immune system, capable of rapidly producing highly specific antibodies upon immunisation, has been used to produce murine monoclonal antibodies (mAbs) via immortalisation and isolation of antibody-secreting cells. These mAbs have had a profound impact in the fields of diagnostics and therapeutics. However, the therapeutic use of murine mAbs is complicated by their immunogenicity. To circumvent this immunogenicity, antibody engineering techniques which render murine mAbs more compatible with the human immune system have been devised. Over the last decade, the technique of antibody phage display, which facilitates the production of human mAbs, has been developed. This review serves as an introduction to antibody engineering, phage display and the development of antibody fragments into viable diagnostic and therapeutic reagents.


Algorithm for prediction of tumour suppressor p53 affinity for binding sites in DNA.
The tumour suppressor p53 is a transcription factor that binds DNA in the vicinity of the genes it controls. The affinity of p53 for specific binding sites relative to other DNA sequences is an inherent driving force for specificity, all other things being equal. We measured the binding affinities of systematically mutated consensus p53 DNA-binding sequences using automated fluorescence anisotropy titrations. Based on measurements of the effects of every possible single base-pair substitution of a consensus sequence, we defined the DNA sequence with the highest affinity for full-length p53 and quantified the effects of deviation from it on the strength of protein-DNA interaction. The contributions of individual nucleotides were to a first approximation independent and additive. But, in some cases we observed significant deviations from additivity. Based on affinity data, we constructed a binding predictor that mirrored the existing p53 consensus sequence definition. We used it to search for high-affinity binding sites in the genome and to predict the effects of single-nucleotide polymorphisms in these sites. Although there was some correlation between the K(d) and biological function, the spread of the K(d)s by itself was not sufficient to explain the activation of different pathways by changes in p53 concentration alone.


Studies on the activation of human neutrophil 5-lipoxygenase induced by natural agonists and Ca2+ ionophore A23187.
By using exogenous substrates, activation of human neutrophil 5-lipoxygenase can be investigated independently of the release of endogenous arachidonic acid. We have developed a sensitive assay to measure 5-LO activation which takes advantage of the 5-LO-mediated conversion of 15S-hydroperoxy-5,8,11,13(Z,Z,Z,E)-eicosatetraenoic acid (15-HpETE) into 5S,15S-dihydroxy-6,8,11,13(E,Z,Z,E)-eicosatetraenoic acid (5,15-DiHETE). When resting neutrophils were incubated with low micromolar concentrations of 15-HpETE, a minor dose- and time-dependent formation of 5,15-DiHETE was observed. In contrast, co-addition of 15-HpETE with Ca2+ ionophore A23187 or with the neutrophil agonists platelet-activating factor (PAF), fMetLeuPhe or complement component C5a resulted in a sizeable concentration-dependent synthesis of 5,15-DiHETE, while lyso-PAF and phorbol myristate acetate were without effect on 5,15-DiHETE formation from 15-HpETE. This stimulation of 5,15-DiHETE synthesis by A23187 or by natural agonists was effectively inhibited by MK-886, a compound that has recently been reported to inhibit the A23187-induced translocation of 5-LO to membrane structures. Furthermore, natural-agonist-induced activation of the 5-LO-mediated transformation of 15-HpETE was inhibited by pertussis toxin, indicating the involvement of a GTP-binding protein in the 5-LO activation process.


Effect of thermal water and adjunctive electrotherapy on chronic low back pain: a double-blind, randomized, follow-up study.
The aim of this study was to evaluate the effectiveness of thermal mineral water, compared with tap water in the treatment of low back pain. This randomized, double-blind, controlled, follow-up study included 71 patients who underwent 20-minute daily treatment sessions with medicinal water or with tap water, both at a temperature of 34 degrees C, on 21 occasions. Both groups underwent additional adjunctive electrotherapy. Outcome measures were visual analogue scale scores, Schober's sign, Domján's signs, Oswestry disability and Short Form-36 questionnaire. The study parameters were administered at baseline, immediately after treatment, and after 15 weeks. After treatment, there was a significant improvement in all parameters in the thermal water group. This improvement was still evident after 15 weeks. The improvement in the control group was less substantial compared with baseline values. Comparison of the 2 treatments revealed a statistically significant difference in 3 outcome parameters (visual analogue scale scores III, IV and Schober's index). In the subset of patients who completed the study according to the protocol, the greater efficacy of treatment with thermal water was also confirmed by the other study parameters. In the group treated with thermal water, improvement occurred earlier, lasted longer and was statistically significant.


An integrated genome-wide association analysis on rheumatoid arthritis data.
We propose a nonparametric association analysis combining both family and unrelated case-control genotype data. Under the assumption of Hardy-Weinberg equilibrium, we formed an affected group to compare with a group of unaffecteds.Comparison with traditional case-control chi-square test and transmission-disequilibrium test shows that this new approach has noticeably improved power. All analysis was based on the simulated rheumatoid arthritis data provided by Genetic Analysis Workshop 15. In the situation of population stratification, we also suggest an approach to update the genotype data using principal components. However, the Genetic Analysis Workshop 15 simulation data does not simulate population stratification. All analysis was done without knowledge of the answers.


Silicon coordination and speciation changes in a silicate liquid at high pressures.
Coordination and local geometry around Si cations in silicate liquids are of primary importance in controlling the chemical and physical properties of magmas. Pressure-induced changes from fourfold to sixfold coordination of Si in silicate glass samples quenched from liquids has been detected with (29)Si magic-angle spinning nuclear magnetic resonance spectrometry. Samples of Na(2)Si(2)O(5) glass quenched from 8 gigapascals and 1500 degrees C contained about 1.5 percent octahedral Si, which was demonstrably part of a homogeneous, amorphous phase. The dominant tetrahedral Si speciation in these glasses became disproportionated to a more random distribution of bridging and nonbridging oxygens with increasing pressure.


Evaluation of skills in arthroscopic training based on trajectory and force data.
Objective evaluation of surgical skills is essential for an arthroscopic training system. We asked whether a quantitative assessment of arthroscopic skills using scores, time to completion, instrument tip trajectory data, and force data was valid. We presumed more experienced surgeons would perform better on a simulated arthroscopic procedure than novices, therefore validating the quantitative assessment. Surgical trainees (n = 12), orthopaedic residents (n = 12), and experienced arthroscopic surgeons (n = 6) were tested on a Sawbones knee simulator. Subjects performed a joint inspection and probing task and a partial meniscectomy task. The trajectory data were measured using an electromagnetic motion tracking system and the force data were measured using a force sensor. The experienced group performed both tasks with higher scores and more quickly than the less experienced groups. The path length of the probe and the scissors was substantially shorter and the probe velocity was considerably faster in the experienced group. The trainee group applied substantially stronger forces to the joint during the joint inspection and probing task. Our data suggest a performance assessment using an electromagnetic motion tracking system and a force sensor provides an objective means of evaluating surgical skills in an arthroscopic training system.


Pneumocystis pneumonia: clinical presentation and diagnosis in patients with and without acquired immune deficiency syndrome.
Pneumocystis carinii causes severe pneumonia in immunocompromised hosts. Although this most commonly occurs in patients with the acquired immunodeficiency syndrome (AIDS), other groups of immunocompromised patients without AIDS are also at risk for P. carinii pneumonia. These patients have solid or hematologic malignancies, organ transplantation, or inflammatory conditions requiring chronic immunosuppressive drugs, particularly corticosteroids. There are important differences in the clinical presentation of P. carinii pneumonia in patients with and without AIDS. P. carinii causes an acute fulminate pneumonia in patients without AIDS while patients with AIDS have more insidious involvement. The organism burden and lung inflammatory response are markedly different between these groups, contributing to substantial differences in clinical presentation, outcome, and mortality. The diagnosis of P. carinii pneumonia remains challenging for primary care physicians and specialists alike. The specific diagnosis of P. carinii pneumonia requires demonstration of the organism from a clinically relevant source, such as sputum, bronchoalveolar lavage fluid, or lung tissue. Alternative methods to diagnose P. carinii pneumonia are currently investigational and are an active area of research. The rapid and specific diagnosis of P. carinii pneumonia allows institution of specific treatment and improvement in patient outcome.


A comparison between behavioral and verbal report pain assessment tools for use with residents in long term care.
The purpose of this study was twofold: (1) to evaluate four pain assessment tools for use with long-term care (LTC) residents who were both able and not able to verbally report their pain; and (2) to assess whether pain behaviors displayed by LTC residents vary as a function of ability to self-report pain. We examined the differences between these two groups of residents in terms of specific pain behaviors assessed through the Pain Assessment Checklist for Seniors with Limited Ability to Communicate (PACSLAC) and Pain Assessment in the Communicatively Impaired (PACI). We also examined the interrater reliability, and concurrent and construct validity of these two behavioral observation tools and the concurrent and construct validity of the two verbal report tools. Data were collected from a convenience sample of 338 residents from six LTC homes. The interrater reliabilities for the two behavioral observation tools were good, and concurrent validity was supported for all four pain assessment tools. Pain behaviors, as assessed by the PACSLAC and PACI, varied as a function of resident ability to verbally report pain. Residents with inability to self-report pain are more nonverbally responsive, although certain behaviors (such as guarding and touching the affected area) are seen more frequently in those capable of self-report. Our data also provide psychometric support for the assessment methods used in this study.


Objective measurement of resolution in film images.
It has been found experimentally that a correlation exists between visually determined resolution limits using four-bar patterns recorded on film and the measured intensity of laser light diffracted from a grating pattern recorded on the film near the four-bar patterns. This correlation has been established for one-lens type and film combination. A practical device has been constructed that reads resolution and is suitable for a production test of a lens-film system.


Analysis of Nicotiana tabacum PIN genes identifies NtPIN4 as a key regulator of axillary bud growth.
The plant-specific PIN-FORMED (PIN) auxin efflux proteins have been well characterized in many plant species, where they are crucial in the regulation of auxin transport in various aspects of plant development. However, little is known about the exact roles of the PIN genes during plant development in Nicotiana species. This study investigated the PIN genes in tobacco (Nicotiana tabacum) and in two ancestral species (Nicotiana sylvestris and Nicotiana tomentosiformis). Genome-wide analysis of the N. tabacum genome identified 20 genes of the PIN family. An in-depth phylogenetic analysis of the PIN genes of N. tabacum, N. sylvestris and N. tomentosiformis was conducted. NtPIN4 expression was strongly induced by the application of exogenous indole-3-acetic acid (IAA), but was downregulated by the application of ABA, a strigolactone analogue, and cytokinin, as well as by decapitation treatments, suggesting that the NtPIN4 expression level is likely positively regulated by auxin. Expression analysis indicated that NtPIN4 was highly expressed in tobacco stems and shoots, which was further validated through analysis of the activity of the NtPIN4 promoter. We used CRISPR-Cas9 technology to generate mutants for NtPIN4 and observed that both T0 and T1 plants had a significantly increased axillary bud growth phenotype, as compared with the wild-type plants. Therefore, NtPIN4 offers an opportunity for studying auxin-dependent branching processes.


4-Methylumbelliferone inhibits tumour cell growth and the activation of stromal hyaluronan synthesis by melanoma cell-derived factors.
There is a close correlation between tumour progression and hyaluronan production, either by tumour cells or by stromal cells that are stimulated by tumour-derived factors. Inhibition of tumour stimulation of fibroblast hyaluronan may suppress tumour growth and invasion. To examine the effect of the hyaluronan synthesis inhibitor 4-methylumbelliferone (4-MU) on the growth of and hyaluronan synthesis by fibroblasts and C8161 and MV3 melanoma cell lines, invasion, and inhibition of tumour cell-derived factor activation of fibroblasts. Effects of 4-MU on growth and hyaluronan synthesis by fibroblasts and melanoma cells were examined in monolayer culture and fibroblast-contracted collagen lattices, and their effects on the growth and invasion of tumour cells into collagen lattices were also studied. 4-MU caused a dose-dependent growth inhibition of fibroblast and melanoma cells with maximum inhibition at 0·5 mmol L(-1) 4-MU. At this dose, 4-MU inhibited (3) H-glucosamine incorporation into fibroblast glycosaminoglycans by 52%, and hyaluronan synthesis by 64%. The relative inhibition was more pronounced when fibroblasts were stimulated with C8161 melanoma cell-conditioned medium. 4-MU reduced the level of hyaluronan in fibroblast-contracted collagen lattices, and inhibited both the growth on and invasion into the lattices by melanoma cells. This growth inhibition appears to be predominantly independent of inhibition of hyaluronan synthesis. The effect on growth inhibition was reversible, and 4-MU had no effect on apoptosis. 4-MU is a potent inhibitor of hyaluronan synthesis, induction of stromal hyaluronan accumulation by tumour cells, and fibroblast and melanoma cell proliferation, and results suggest that 4-MU may have potential as a tumour cell anti-invasive and antiproliferative agent.


[Infrared study on molecular structure and orientation in Langmuir-Blodgett films of chiral LCs M1].
The LB films of chiral LCs M1 were investigated by FTIR transmission and reflection absorption spectra. CaF2 and vacuum deposition gold plate were used as solid substrate. The LB films can be transferred onto the solid substrate successfully. The orientation of LB films was analysis by comparing the FTIR transmission and reflection absorbance spectroscopy. The aromatic cores part which including the C-O bond was almost perpendicular the substrate surface, the alkyl chain and C=O part has a certain angle titled onto the substrate.


What can we learn about cardioprotection from the cardiac mitochondrial proteome?
This review will summarize proteomic methods that are useful in studying the role of mitochondria in cardioprotection. The strengths and weaknesses of some of the different approaches are discussed. We focus on the cardiac mitochondrial proteome with emphasis on changes associated with cell death and protection, and we summarize how proteomic data have contributed to addressing the role of mitochondria in cardioprotection.


Effects of a history of differential reinforcement on preference for choice.
The effects of a history of differential reinforcement for selecting a free-choice versus a restricted-choice stimulus arrangement on the subsequent responding of 7 undergraduates in a computer-based game of chance were examined using a concurrent-chains arrangement and a multiple-baseline-across-participants design. In the free-choice arrangement, participants selected three numbers, in any order, from an array of eight numbers presented on the computer screen. In the restricted-choice arrangement, participants selected the order of three numbers preselected from the array of eight by a computer program. In initial sessions, all participants demonstrated no consistent preference or preference for restricted choice. Differential reinforcement of free-choice selections resulted in increased preference for free choice immediately and in subsequent sessions in the absence of programmed differential outcomes. For 5 participants, changes in preference for choice were both robust and lasting, suggesting that a history of differential reinforcement for choice may affect preference for choice.


Gingival cyst of the adult: a case report.
The gingival cyst of the adult is an uncommon odontogenic cyst of developmental origin that is believed to represent the soft tissue counterpart of the lateral periodontal cyst. This article describes a case of gingival cyst of the adult, located in the alveolar mucosa on the facial aspect of the mandibular lateral incisor-canine region. The lesion was nodular with a sessile base, nonulcerated, and nonpainful; it was normal in color and measured 5.0 mm in diameter. An excisional biopsy was performed and surgical exploration revealed osseous surface erosion. Histopathological analysis confirmed the clinical hypothesis.


Functional Interaction Between Drosophila Olfactory Sensory Neurons and Their Support Cells.
Insects detect volatile chemicals using antennae, which house a vast variety of olfactory sensory neurons (OSNs) that innervate hair-like structures called sensilla where odor detection takes place. In addition to OSNs, the antenna also hosts various support cell types. These include the triad of trichogen, tormogen, and thecogen support cells that lie adjacent to their respective OSNs. The arrangement of OSN supporting cells occurs stereotypically for all sensilla and is widely conserved in evolution. While insect chemosensory neurons have received considerable attention, little is known about the functional significance of the cells that support them. For instance, it remains unknown whether support cells play an active role in odor detection, or only passively contribute to homeostasis, e.g., by maintaining sensillum lymph composition. To investigate the functional interaction between OSNs and support cells, we used optical and electrophysiological approaches in Drosophila. First, we characterized the distribution of various supporting cells using genetic markers. By means of an ex vivo antennal preparation and genetically-encoded Ca2+ and K+ indicators, we then studied the activation of these auxiliary cells during odor presentation in adult flies. We observed acute responses and distinct differences in Ca2+ and K+ fluxes between support cell types. Finally, we observed alterations in OSN responses upon thecogen cell ablation in mature adults. Upon inducible ablation of thecogen cells, we notice a gain in mechanical responsiveness to mechanical stimulations during single-sensillum recording, but a lack of change to the neuronal resting activity. Taken together, these results demonstrate that support cells play a more active and responsive role during odor processing than previously thought. Our observations thus reveal that support cells functionally interact with OSNs and may be important for the extraordinary ability of insect olfactory systems to dynamically and sensitively discriminate between odors in the turbulent sensory landscape of insect flight.


Spacetime Symmetries and Conformal Data in the Continuous Multiscale Entanglement Renormalization Ansatz.
The generalization of the multiscale entanglement renormalization ansatz (MERA) to continuous systems, or cMERA [Haegeman et al., Phys. Rev. Lett. 110, 100402 (2013)PRLTAO0031-900710.1103/PhysRevLett.110.100402], is expected to become a powerful variational ansatz for the ground state of strongly interacting quantum field theories. In this Letter, we investigate, in the simpler context of Gaussian cMERA for free theories, the extent to which the cMERA state |Ψ^{Λ}⟩ with finite UV cutoff Λ can capture the spacetime symmetries of the ground state |Ψ⟩. For a free boson conformal field theory (CFT) in 1+1 dimensions, as a concrete example, we build a quasilocal unitary transformation V that maps |Ψ⟩ into |Ψ^{Λ}⟩ and show two main results. (i) Any spacetime symmetry of the ground state |Ψ⟩ is also mapped by V into a spacetime symmetry of the cMERA |Ψ^{Λ}⟩. However, while in the CFT, the stress-energy tensor T_{μν}(x) (in terms of which all the spacetime symmetry generators are expressed) is local, and the corresponding cMERA stress-energy tensor T_{μν}^{Λ}(x)=VT_{μν}(x)V^{†} is quasilocal. (ii) From the cMERA, we can extract quasilocal scaling operators O_{α}^{Λ}(x) characterized by the exact same scaling dimensions Δ_{α}, conformal spins s_{α}, operator product expansion coefficients C_{αβγ}, and central charge c as the original CFT. Finally, we argue that these results should also apply to interacting theories.


Multivariate autoregressive feature extraction and the recognition of multichannel waveforms.
It is proposed that the autoregressive coefficient matrices appearing in a multivariate autoregressive model fitting may be used for feature extraction purposes in problems concerning recognition of multichannel waveforms. It is demonstrated how the information contained in the autoregressive parameters may be further compressed by applying the ordinary or a modified Karhunen-Loeve expansion. The feature extraction procedures are illustrated on a large data base of seismic wave traces originating from shallow earthquakes and underground nuclear explosions. The results obtained (using a multivariate Gaussian classification algorithm) suggest that the combined autore-gressive/Karhunen-Loeve method has a considerably larger discrimination potential than the more conventional seismic discriminants.


Changes in renal medullary transport proteins during uncontrolled diabetes mellitus in rats.
We tested whether the abundance of transport proteins involved in the urinary concentrating mechanism was altered in rats with uncontrolled diabetes mellitus (DM). Rats were injected with streptozotocin and killed 5, 10, 14, or 20 days later. Blood glucose in DM rats was 300-450 mg/dl (control: 70-130 mg/dl). Urine volume increased in DM rats from 41 +/- 7 ml/100 g body wt (BW) at 5 days to 69 +/- 3 ml/100 g BW at 20 days (control: 9 +/- 1). Urine osmolality of DM rats decreased at 5 days DM and remained low at 20 days. UT-A1 urea transporter protein in the inner medullary (IM) tip was 55% of control in 5-day DM rats but increased to 170, 220, and 280% at 10, 14, and 20 days DM, respectively, due to an increase in the 117-kDa glycoprotein form. UT-A1 in the IM base was increased to 325% of control at 5 days DM with no further increase at 20 days. Aquaporin-2 (AQP2) increased to 290% in the IM base at 5 days DM and 150% in the IM tip at 10 days; both showed no further increase at 20 days. NKCC2/BSC1 increased to 240% in outer medulla at 20 days DM, but not at 5 or 10 days. UT-B and ROMK were unchanged at any time point. The increases in UT-A1, AQP2, and NKCC2/BSC1 proteins during uncontrolled DM would tend to limit the loss of fluid and solute during uncontrolled diabetes.


Can Transcranial Direct-Current Stimulation Alone or Combined With Cognitive Training Be Used as a Clinical Intervention to Improve Cognitive Functioning in Persons With Mild Cognitive Impairment and Dementia? A Systematic Review and Meta-Analysis.
Background: Transcranial direct-current stimulation (tDCS) facilitates cognitive improvement in healthy and pathological populations. It has been increasingly used in cases of mild cognitive impairment (MCI) and dementia. Our research question is: Can tDCS serve as a clinical intervention for improving the cognitive functions of persons with MCI (PwMCI) and dementia (PwD)? Objective: This systematic review evaluated the evidence to determine the efficacy of tDCS in improving cognitive outcomes in PwD and PwMCI. Methods: A systematic review was conducted of studies published up to November 2017 involving tDCS in cases of MCI and dementia. Studies were ranked according to the level of evidence (Oxford Center for Evidence-Based Medicine) and assessed for methodological quality (Risk of Bias Tool in the Cochrane Handbook for Systematic Reviews of Interventions). Data was extracted on all protocol variables to establish a reference framework for clinical interventions. Different modalities, tDCS alone or combined with cognitive training, compared with sham tDCS were examined in both short and long-term effects. Four randomized control trials (RCTs) with memory outcomes were pooled using the fixed-effect model for the meta-analysis. Results: Twelve studies with 195 PwD and four with 53 PwMCI met the inclusion criteria. Eleven articles were ranked as Level 1b. The results on the meta-analysis on pooled effects of memory indicated a statistically significant medium effect size of 0.39 (p = 0.04) for immediate effects. This improvement was not maintained in the long term 0.15 (p = 0.44). Conclusion: tDCS improves memory in PwD in the short term, it also seems to have a mild positive effect on memory and language in PwMCI. However, there is no conclusive advantage in coupling tDCS with cognitive training. More rigorous evidence is needed to establish whether tDCS can serve as an evidence-based intervention for both populations.


Development and validation of an LC-MS/MS method for the determination of AZD7648 in rat plasma: Application to a pharmacokinetic study.
AZD7648 is a potent DNA-PK inhibitor that is being developed for the treatment of ovarian cancer. The study aimed to develop a simple and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine the concentration of AZD7648 in rat. AZD7648 was extracted from plasma by acetonitrile-mediated protein precipitation. The quantification was performed on a Thermo Vantage TSQ mass spectrometer with ibrutinib as an internal standard. A Waters Acquity UPLC BEH C18 column combined with 0.1% aqueous formic acid and acetonitrile was employed for chromatographic separation. The precursor-to-product ion transitions were m/z 421.2 > 337.2 and m/z 441.2 > 138.1 for AZD7648 and internal standard, respectively. This method was successfully validated according to the US Food and Drug Administration guidance. The calibration curve was linear over the concentration range of 0.5-1,000 ng/ml with correlation coefficient >0.999. The precision expressed as the coefficient of variation was <8.09%, while the accuracy expressed as relative error ranged from -10.00 to 9.08%. The mean recovery was >94.49%. AZD7648 was stable in rat plasma after storage under certain conditions. The validated method was demonstrated to be selective, sensitive and reliable, and has been successfully applied to the pharmacokinetic study of AZD7648 in rat plasma after oral and intravenous administration (1 mg/kg).


[The construction of collective portfolios in traditional curriculums: an innovative approach in teaching-learning].
Education to promote health has traditionally been based on knowledge transmission methodologies. However, the current scenario calls for the training of professionals with a critical-reflective profile, who are able to work in teams. We present the report of an innovative experience using the construction of collective portfolios as instruments of learning, changing attitudes and training of undergraduates, in a traditional subject-based curriculum structure context. It is a descriptive exploratory study, with a qualitative-quantitative approach, based on analysis of collective portfolios (n=9), built by Health Policy students, together with an open questionnaire to students who attended the course (n=58) and also the staging of focus groups (n=3). The use of collective portfolios mobilized students in critical and reflective thinking on Brazilian health policy - the Unified Health System - broadening the concept on the health-disease process and practices related to health services, prioritizing teamwork and the active search for knowledge building, stressing the exercise of otherness, resilience and empowerment.


The nucleotide sequences of two leghemoglobin genes from soybean.
We present the complete nucleotide sequences of two leghemoglobin genes isolated from soybean DNA. Both genes contain three intervening sequences in identical positions. Comparison of the coding sequences with known amino-acid sequences of soybean leghemoglobins suggest that the two genes correspond to leghemoglobin C2 and leghemoglobin C3, respectively.


Information: what do consumers want to know?
National statistics indicate the potential demand for health care information will become an increasingly important area of attention in the years ahead. Consumers currently focus on such factors as reputation, accessibility, and cost when making health care decisions. Future consumers will be concerned with the interests, experience, satisfaction, and outcomes of health professionals, facilities, and plans. Current public dissemination efforts will create additional demand for more and better health care information.


The glycolipid asialo GM1 as a new differentiation antigen of fetal thymocytes.
Antibody directed to the neutral glycolipid "asialo GM1" was found to react with the majority of thymus lymphoid cells lacking characteristic T cell markers in mice at an early embryonic stage (13 days of gestation). The proportion of these asialo GM1-positive cells (asialo GM1+ cells) decreased strikingly thereafter, contrasting with an increased fraction of lymphocytes possessing T cell surface markers such as Thy-1, Lyt-1, and Lyt-2. After about 18 gestational days, only a few asialo GM1+ cells were detected in the thymus as well as in other lymphoid organs. Double-staining analysis of the embryonic thymocytes (13 days of gestation) with anti-asialo GM1 and anti-Thy-1 demonstrated that thymocytes stained with anti-asialo GM1 did not react with anti-Thy-1, and vice versa. Morphologic examination by immunoelectronmicroscopy demonstrated that these asialo GM1+ cells were mainly composed of immature, large lymphoid cells having large nucleoli and relatively abundant cytoplasm with many polysomes. These results suggest that asialo GM1 is present on very early thymocytes and is lost as the mature murine T cell protein antigens Thy-1, Lyt-1, and Lyt-2 develop on these cells. The antibody to this glycolipid is a useful tool for studying embryonic thymic differentiation.


GlcNAc is a mast-cell chromatin-remodeling oncometabolite that promotes systemic mastocytosis aggressiveness.
Systemic mastocytosis (SM) is a KIT-driven hematopoietic neoplasm characterized by the excessive accumulation of neoplastic mast cells (MCs) in various organs and, mainly, the bone marrow (BM). Multiple genetic and epigenetic mechanisms contribute to the onset and severity of SM. However, little is known to date about the metabolic underpinnings underlying SM aggressiveness, which has thus far impeded the development of strategies to leverage metabolic dependencies when existing KIT-targeted treatments fail. Here, we show that plasma metabolomic profiles were able to discriminate indolent from advanced forms of the disease. We identified N-acetyl-d-glucosamine (GlcNAc) as the most predictive metabolite of SM severity. High plasma levels of GlcNAc in patients with advanced SM correlated with the activation of the GlcNAc-fed hexosamine biosynthesis pathway in patients BM aspirates and purified BM MCs. At the functional level, GlcNAc enhanced human neoplastic MCs proliferation and promoted rapid health deterioration in a humanized mouse model of SM. In addition, in the presence of GlcNAc, immunoglobulin E-stimulated MCs triggered enhanced release of proinflammatory cytokines and a stronger acute response in a mouse model of passive cutaneous anaphylaxis. Mechanistically, elevated GlcNAc levels promoted the transcriptional accessibility of chromatin regions that contain genes encoding mediators of receptor tyrosine kinases cascades and inflammatory responses, thus leading to a more aggressive phenotype. Therefore, GlcNAc is an oncometabolite driver of SM aggressiveness. This study suggests the therapeutic potential for targeting metabolic pathways in MC-related diseases to manipulate MCs effector functions.


Cardioprotective Function of Green Rooibos (Aspalathus linearis) Extract Supplementation in Ex Vivo Ischemic Prediabetic Rat Hearts.
Diabetic patients develop ischemic heart disease and strokes more readily. Following an ischemic event, restoration of blood flow increases oxidative stress resulting in myocardial damage, termed ischemia/reperfusion injury. Aspalathus linearis (rooibos), rich in the antioxidant phenolic compound aspalathin, has been implicated as cardioprotective against ischemia/reperfusion injury with undefined mechanism in control rats. Primarily, the therapeutic potential of Afriplex green rooibos extract to prevent ischemia/reperfusion injury in cardiovascular disease-compromised rats was investigated. Additionally, Afriplex Green rooibos extract's cardioprotective signaling on metabolic markers and stress markers was determined using western blotting. Three hundred male Wistar rats received either 16-wk standard diet or high-caloric diet. During the final 6 wk, half received 60 mg/kg/day Afriplex green rooibos extract, containing 12.48% aspalathin. High-caloric diet increased body weight, body fat, fasting serum triglycerides, and homeostatic model assessment of insulin resistance - indicative of prediabetes. High-caloric diet rats had increased heart mass, infarct size, and decreased heart function. Afriplex green rooibos extract treatment for 6 wk lowered pre-ischemic heart rate, reduced infarct size, and improved heart function pre- and post-ischemia, without significantly affecting biometric parameters. Stabilized high-caloric diet hearts had decreased insulin independence via adenosine monophosphate activated kinase and increased inflammation (p38 mitogen-activated protein kinase), whereas Afriplex green rooibos extract treatment decreased insulin dependence (protein kinase B) and conferred anti-inflammatory effect. After 20 min ischemia, high-caloric diet hearts had upregulated ataxia-telangiectasia mutated kinase decreased insulin independence, and downregulated insulin dependence and glycogen synthase kinase 3 β inhibition. In contrast, Afriplex green rooibos extract supplementation downregulated insulin independence and inhibited extracellular signal-regulated kinase 1 and 2. During reperfusion, all protective signaling was decreased in high-caloric diet, while Afriplex green rooibos extract supplementation reduced oxidative stress (c-Jun N-terminal kinases 1 and 2) and inflammation. Taken together, Afriplex green rooibos extract supplementation for 6 wk preconditioned cardiovascular disease-compromised rat hearts against ischemia/reperfusion injury by lowering inflammation, oxidative stress, and heart rate.


Antimicrobial susceptibilities of Escherichia coli isolates as agents of community-acquired urinary tract infection (2008-2014).
Urinary tract infections (UTIs) are among the most frequently seen community-acquired infections worldwide. E. coli causes 90% of urinary system infections. To guide the empirical therapy, the resistance pattern of E. coli responsible for community-acquired UTI was evaluated throughout a seven-year period in this study. The urine cultures of patients with urinary tract infections admitted to outpatient clinics between 1(st) January 2008 and 31(st) December 2014 were analyzed. Presence of ≥10(5) colony-forming units/mL in urine culture media was considered as significant for UTI. Isolated bacteria were identified by standard laboratory techniques or automated system VITEK2 (BioMerieux, France) and BD PhoenixTM 100 (BD, USA), as required. Antibiotic susceptibility testing was performed by Kirby-Bauer disk diffusion method using Clinical Laboratory Standard Institute (CLSI) criteria. A total of 13281 uropathogens were isolated. Overall E. coli accounted for 8975 (67%) of all isolates. Resistance rates of E. coli to antimicrobial agents was demonstrated to be as follows: ampicillin 66.9%, cefazolin 30.9%, cefuroxime 30.9%, ceftazidime 14.9%, cefotaxime 28%, cefepime 12%, amoxicillin-clavulanic acid 36.9%, trimethoprim-sulfamethoxazole (TMP-SXT) 20%, ciprofloxacin 49.9%, amikacin 0.3%, gentamycin 24%, nitrofurantoin 0.9%, and fosfomycin 4.3%. There was no resistance to imipenem nor meropenem. The frequency of ESBL-producing E. coli strains was 24%. It is concluded that fosfomycin and nitrofurantoin are appropriate empirical therapy for community-acquired UTI empirical therapy, but the fluoroquinolones and the TMP-SXT shall not be used in the emprical treatment of UTI at this stage. In conclusion, as resistance rates show regional differences, it is necessary to regularly examine regional resistance rates to determine the appropriate empiric antibiotic treatment and national antibiotic usage policies must be reorganized according to data obtained from these studies.


Pulmonary Thrombosis: A Clinical Pathological Entity Distinct from Pulmonary Embolism?
Since Virchow's autopsy studies in the mid-1800s, it has generally been believed that pulmonary embolism (PE) originated from the embolization of fibrin fragments from a deep venous thrombosis (DVT). However, a DVT is often not found in patients with PE (up to 50% of cases). Could fibrin form in the pulmonary vessels without coming from the periphery? In this review, the authors will try to support the hypothesis that a pulmonary thrombosis (PT) may develop. They will do so through different clinical models related to some pathological conditions such as pneumonia, chronic obstructive pulmonary disease (COPD), and asthma, all of which show a close relationship between local inflammation and activation of blood coagulation, two defensive systems that may lead to fibrin deposition in the lungs, thus recognizing the possibility that PT may be a newly recognized entity. An increased risk for PE has been demonstrated in these conditions. Sickle cell disease and assisted reproductive technologies are other very different conditions in which an increased risk for PE has been found. Gaucher's disease is a rare hereditary condition in which the hemostatic system could have a role in the pathogenesis of pulmonary hypertension, which complicates the course of the disease. In particular, the increased risk for PT, common to all these conditions, deserves attention when a patient presents with sudden dyspnea, an unexpected COPD exacerbation, or severe sudden asthmatic dyspnea. As a consequence, prediction scores for venous thromboembolism could be revised.


Strategic information systems.
Successful transformation to a regional health care delivery system, rather than independent group practices and hospitals, will depend on the strategic use of information technology. Frank T. Sample, president and CEO of Phamis Inc., writes that information systems will be essential for achieving business and care objectives.


Visual input evokes transient and strong shunting inhibition in visual cortical neurons.
The function and nature of inhibition of neurons in the visual cortex have been the focus of both experimental and theoretical investigations. There are two ways in which inhibition can suppress synaptic excitation. In hyperpolarizing inhibition, negative and positive currents sum linearly to produce a net change in membrane potential. In contrast, shunting inhibition acts nonlinearly by causing an increase in membrane conductance; this divides the amplitude of the excitatory response. Visually evoked changes in membrane conductance have been reported to be nonsignificant or weak, supporting the hyperpolarization mode of inhibition. Here we present a new approach to studying inhibition that is based on in vivo whole-cell voltage clamping. This technique allows the continuous measurement of conductance dynamics during visual activation. We show, in neurons of cat primary visual cortex, that the response to optimally orientated flashed bars can increase the somatic input conductance to more than three times that of the resting state. The short latency of the visually evoked peak of conductance, and its apparent reversal potential suggest a dominant contribution from gamma-aminobutyric acid ((GABA)A) receptor-mediated synapses. We propose that nonlinear shunting inhibition may act during the initial stage of visual cortical processing, setting the balance between opponent 'On' and 'Off' responses in different locations of the visual receptive field.


Electrophysiological findings in diphenyl poisoning.
The fungistatic agent diphenyl caused fatal poisoning with signs of neurotoxicity in a worker in a Finnish paper mill. This initiated a neurophysiological study of 24 workers occupationally exposed to diphenyl. Ten men showed EEG abnormalities, mainly diffuse or generalized ones. The abnormalities persisted on re-examination one and two years later. Nine subjects had EMG abnormalities; seven also exhibited fibrillations in some muscles. One subject showed a long rhythmic series of fasciculations similar to the spontaneous activity described in infantile spinal muscular atrophy. Nerve conduction velocity, especially that of slower motor fibres, was reduced in several cases. Electroneuromyographic abnormalities also persisted on re-examination. Although diphenyl is considered a comparatively safe chemical, it showed evidence of neurotoxicity when workers were exposed to concentrations in excess of the presently accepted threshold limits. Electrophysiological methods should be applied for the early detection of occupational hazards.


Effect of harvesting index on browning reaction and changes of tissue structure in santol fruits.
This investigation was carried out to justify the relationship between harvesting ages of santol fruit, browning reaction and other concerning data. Growers of santol plantations in Thailand have always accustomed to problems on inconsistency in qualities of santol fruits var. Pui Fai due to inappropriate harvesting index. Thus these encourage to decrease marketable qualities of fruits and short shelf-life particularly browning discoloration. In order to investigate adequate information for growers of the santol orchard plants, it is important to carry out experiments on Polyphenol Oxidase (PPO) activity and changes of tissue structure by separating fruit into three parts: peel, flesh and seed of santol fruit var. Pui Fai harvested at four stages: 100, 115, 130 and 145 Days After Full bloom (DAF). A factorial in randomized complete block design with five replications was used. In each replication ten fruits were used. This investigation was carried out during the period of October 2006 to March 2007 at The Department of Agricultural Technology, Mahasarakham University, Northeast Thailand. The results showed that PPO activity which related to browning reaction changed with harvesting stage. The highest PPO activities were obtained and highly significant increased in mature fruit at 130 and 145 DAF. When the fruits were ripening at 145 DAF, PPO activities from peel, flesh and seed had no significant differences and high levels by 160.76-184.44, 158.88-180.76 and 154.52-181.08 U mg(-1) fresh weight, respectively. While cross-section study in peel and flesh of santol fruit var. Pui Fai at different ages indicated that immature fruits (100 and 115 DAF) showed small cells arranged one to two layers in epidermis. Trichome like-hair also appeared on this layer. When santol fruits grew through 145 DAF, epidermis converted to periderm and trichome disappeared. In addition, parenchyma cells in flesh accumulated several substances in form of druse with the fruit age increment. These high PPO activities and fruit tissue conversions of peel and flesh in mature fruit may led to be more sensitive to browning reaction.


The effects of resveratrol on cyclooxygenase-1 and -2, nuclear factor kappa beta, matrix metalloproteinase-9, and sirtuin 1 mRNA expression in hearts of streptozotocin-induced diabetic rats.
Resveratrol (RSV) has a beneficial role in the prevention of diabetes and alleviates some diabetic complications, such as cardiomyopathy. We investigated cyclooxygenase-1 (COX-1), COX-2, nuclear factor κB (NF-κB), matrix metalloproteinase-9 (MMP-9), and sirtuin 1 (SIRT1) mRNA expression levels in heart tissue after RSV treatment in streptozotocin (STZ)-induced diabetic rats. After induction of chronic diabetes with STZ, 10 mg RSV/kg per day was administered to DM and DM+RSV groups for four weeks. At the end of the experiment, all rats were sacrificed and heart tissues were stored at -80°C; mRNA expression levels of COX-1, COX-2, NF-κB, MMP-9, and SIRT1 genes were analyzed with quantitative real-time PCR. We did not find any significant effect of RSV on MMP-9, COX-1, COX-2, or NF-κB mRNA levels among the groups. However, SIRT1 mRNA levels decreased in the DM group compared to controls and increased in the DM+RSV group when compared to the DM group. SIRT1 is activated by RSV treatment in diabetic heart tissue. Activation of SIRT1 by RSV may lead to a new therapeutic approach for diabetic heart tissue. We conclude that RSV treatment can alleviate heart dysfunction by inhibiton of inflammatory gene expression such as SIRT1.


Infection Pattern of Neutropenic Patients in Post-chemotherapy Phase of Acute Leukemia Treatment.
Neutropenia following chemotherapy regimens in leukemia patients is of major concern since it makes these patients vulnerable to infections. If we can identify which germs are causing these infections, they can be annihilated or, at least, the most appropriate antibiotic therapy can be started immediately, even before we have the results of the culture. This retrospective multi-center study took place in 2012 and included patients with acute leukemia who had already undergone chemotherapy and who had been febrile for at least 16 hours. In order to assess the type of infection, different environments were chosen and the results were compared by t-test and χ(2) tests. This study took place in four hospitals in Tehran and Ahwaz, Iran. The study population was made up of 89 patients: 37 with acute lymphoblastic leukemia and 52 with acute myeloid leukemia. The results revealed that blood was the most common site of infection. From all our positive cultures, it was seen that 85.4% of them had gram-negative bacteria with a dominance of E. coli of 25.8% over the other colonies. Also, antibiograms revealed the sensitivity of almost all the gram-negatives to amino glycosides. In contrast with most of the literature, in our patients, gram-negatives are the most common cause of infection and, therefore, administering amino glycosides would be the safest antibiotic therapy to prescribe before culture results are available.


Biallelic truncation variants in ATP9A are associated with a novel autosomal recessive neurodevelopmental disorder.
Intellectual disability (ID) is a highly heterogeneous disorder with hundreds of associated genes. Despite progress in the identification of the genetic causes of ID following the introduction of high-throughput sequencing, about half of affected individuals still remain without a molecular diagnosis. Consanguineous families with affected individuals provide a unique opportunity to identify novel recessive causative genes. In this report, we describe a novel autosomal recessive neurodevelopmental disorder. We identified two consanguineous families with homozygous variants predicted to alter the splicing of ATP9A which encodes a transmembrane lipid flippase of the class II P4-ATPases. The three individuals homozygous for these putatively truncating variants presented with severe ID, motor and speech impairment, and behavioral anomalies. Consistent with a causative role of ATP9A in these patients, a previously described Atp9a-/- mouse model showed behavioral changes.


Antinociceptive Effect of the Essential Oil of Schinus terebinthifolius (female) Leaves on Adult Zebrafish (Danio rerio).
Schinus terebinthifolius Raddi (Anacardiaceae) is popularly known in Brazil as aroeira-da-praia and has pharmacological use as an astringent, antidiarrheal, anti-inflammatory, depurative, diuretic, and antifebrile agent. Although the neuropathic antinociceptive potential of S. terebinthifolius fruits has already been investigated, this study is the first one to analyze the acute antinociceptive effect of the essential oil of S. terebinthifolius (female) leaves (EOFSt) on adult zebrafish. EOFSt was submitted to antioxidant activity evaluation by two methods (ferrous ion-chelating capacity [FIC] and β-carotene). The animals (n = 6/group) were treated orally (20 μL) with EOFSt (0.1, 0.5, or 1.0 mg/mL) or vehicle (0.9% sodium chloride [NaCl]; 20 μL), and submitted to nociception (formalin, cinnamaldehyde, capsaicin, glutamate, acidic saline, and hypertonic saline). Possible neuromodulation mechanisms, as well motor alterations and toxicity were also evaluated. In the FIC assay, EOFSt showed ferrous ion-chelating capacity in ∼40% to 90%. Regarding the β-carotene bleaching assay, EOFSt showed inhibition in a 58% to 80% range. Oral administration of EOFSt showed no acute toxicity and did not alter the locomotor system of aZF, and reduced the nociceptive behavior in all tested models. These effects of EOFSt were significantly similar to those of morphine, used as a positive control. The antinociceptive effect of EOFSt was inhibited by naloxone, L-NAME, ketamine, camphor, ruthenium red, and amiloride. The antinociceptive effect of the EOFSt cornea was inhibited by capsazepine. EOFSt has the pharmacological potential for acute pain treatment and this effect is modulated by the opioid system, NMDA receptors, and transient receptor potential ankyrin 1 (TRPA1), transient receptor potential vanilloid 1 (TRPV1), and acid-sensing ion channels. The EOFSt also has the pharmacological potential for corneal pain treatment and this effect is modulated by the TRPV1 channel.


Identification of a synergistic interaction between endothelial cells and retinal pigment epithelium.
The retinal pigment epithelium located between the neurosensory retina and the choroidal vasculature is critical for the function and maintenance of both the photoreceptors and underlying capillary endothelium. While the trophic role of retinal pigment epithelium on choroidal endothelial cells is well recognized, the existence of a reciprocal regulatory function of endothelial cells on retinal pigment epithelium cells remained to be fully characterized. Using a physiological long-term co-culture system, we determined the effect of retinal pigment epithelium-endothelial cell heterotypic interactions on cell survival, behaviour and matrix deposition. Human retinal pigment epithelium and endothelial cells were cultured on opposite sides of polyester transwells for up to 4 weeks in low serum conditions. Cell viability was quantified using a trypan blue assay. Cellular morphology was evaluated by H&E staining, S.E.M. and immunohistochemistry. Retinal pigment epithelium phagocytic function was examined using a fluorescent bead assay. Gene expression analysis was performed on both retinal pigment epithelium and endothelial cells by quantitative PCR. Quantification of extracellular matrix deposition was performed on decellularized transwells stained for collagen IV, fibronectin and fibrillin. Our results showed that presence of endothelial cells significantly improves retinal pigment epithelium maturation and function as indicated by the induction of visual cycle-associated genes, accumulation of a Bruch's membrane-like matrix and increase in retinal pigment epithelium phagocytic activity. Co-culture conditions led to increased expression of anti-angiogenic growth factors and receptors in both retinal pigment epithelium and endothelial cells compared to monoculture. Tube-formation assays confirmed that co-culture with retinal pigment epithelium significantly decreased the angiogenic phenotype of endothelial cells. These findings provide evidence of critical interdependent interactions between retinal pigment epithelium and endothelial cell involved in the maintenance of retinal homeostasis.


Proteomic Biomarkers for the Detection of Endometrial Cancer.
Endometrial cancer is the leading gynaecological malignancy in the western world and its incidence is rising in tandem with the global epidemic of obesity. Early diagnosis is key to improving survival, which at 5 years is less than 20% in advanced disease and over 90% in early-stage disease. As yet, there are no validated biological markers for its early detection. Advances in high-throughput technologies and machine learning techniques now offer unique and promising perspectives for biomarker discovery, especially through the integration of genomic, transcriptomic, proteomic, metabolomic and imaging data. Because the proteome closely mirrors the dynamic state of cells, tissues and organisms, proteomics has great potential to deliver clinically relevant biomarkers for cancer diagnosis. In this review, we present the current progress in endometrial cancer diagnostic biomarker discovery using proteomics. We describe the various mass spectrometry-based approaches and highlight the challenges inherent in biomarker discovery studies. We suggest novel strategies for endometrial cancer detection exploiting biologically important protein biomarkers and set the scene for future directions in endometrial cancer biomarker research.


Prolonged Bartonella bacteremia in cats associated with cat-scratch disease patients.
Recent evidence supports a causal relationship between Bartonella (Rochalimaea) henselae, cat-scratch disease (CSD), and bacillary angiomatosis. Cats appear to be the primary reservoir. Blood from 19 cats owned by 14 patients diagnosed with CSD was cultured. Blood samples from cats owned by veterinary students (n = 25) having no association with CSD or bacillary angiomatosis were cultured as controls. Eighty-nine percent (17 of 19) of cats associated with CSD patients and 28% (7 of 25) of controls were bacteremic with Bartonella species (chi-square = 16.47; P < 0.001). Twenty-three isolates were characterized as B. henselae, while one isolate from the cat of a CSD patient appeared to be a new Bartonella species. Thirteen cats remained culture positive during the ensuing 12-month period. Our results support the conclusion that B. henselae is the predominant species involved in CSD and is transmitted by cats. The incidence of Bartonella bacteremia in control cats suggests that B. henselae bacteremia is prevalent among the domestic cat population in the United States.


[Formation of myoblasts after injuries to skeletal muscle tissue].
The musculus masseter and the tongue in 75 mature rats have been injured by means of a scalpel, carbonate laser ray and ethanol administration. The regeneration process has been studied light and electron microscopically. Myosatellites that are constantly revealed in the skeletal muscle tissue, are the main source of myoblast formation under all the types of the muscle injury. When laser ray and ethanol are applied, myoblasts are also formed by means of segregation of the nucleo-sarcoplasmic territories of the muscle fiber.


Human placental hematopoietic stem cell derived natural killer cells (CYNK-001) mediate protection against influenza a viral infection.
Influenza A virus (IAV) infections are associated with a high healthcare burden around the world and there is an urgent need to develop more effective therapies. Natural killer (NK) cells have been shown to play a pivotal role in reducing IAV-induced pulmonary infections in preclinical models; however, little is known about the therapeutic potential of adoptively transferred NK cells for IAV infections. Here, we investigated the effects of CYNK-001, human placental hematopoietic stem cell derived NK cells that exhibited strong cytolytic activity against a range of malignant cells and expressed high levels of activating receptors, against IAV infections. In a severe IAV-induced acute lung injury model, mice treated with CYNK-001 showed a milder body weight loss and clinical symptoms, which led to a delayed onset of mortality, thus demonstrating their antiviral protection in vivo. Analysis of bronchoalveolar lavage fluid (BALF) revealed that CYNK-001 reduced proinflammatory cytokines and chemokines highlighting CYNK-001's anti-inflammatory actions in viral induced-lung injury. Furthermore, CYNK-001-treated mice had altered immune responses to IAV with reduced number of neutrophils in BALF yet increased number of CD8+ T cells in the BALF and lung compared to vehicle-treated mice. Our results demonstrate that CYNK-001 displays protective functions against IAV via its anti-inflammatory and immunomodulating activities, which leads to alleviation of disease burden and progression in a severe IAV-infected mice model. The potential of adoptive NK therapy for IAV infections warrants clinical investigation.


Smart home energy management and power trading optimization using an enhanced manta ray foraging optimization.
This paper proposes a plan to manage energy consumption in residential areas using the demand response method, which allows electricity users to contribute to the reliability of the power system by controlling their usage. Due to the growing population, the residential sector consumes a significant amount of energy, and the objectives of this study are to lower electricity costs and the peak to average ratio, as well as reduce the amount of imported electricity from the grid. The study aims to maximize profit by properly utilizing renewable energy sources and addressing energy trading. The manta ray foraging optimization (MRFO) and long term memory MRFO (LMMRFO) algorithms are used to solve this problem. Firstly, the validation of the proposed LMMRFO technique is confirmed by seven benchmark functions and compared its results with the results of the well-known optimization algorithms including hunter prey optimization, gorilla troops optimizer, beluga whale optimization, and the original MRFO algorithm. Then, the performance of the LMMRFO is checked on the optimization of smart home energy management. In the suggested approach, a smart home decides whether to purchase or sell electricity from the commercial grid based on the cost, demand, and production of electricity from its own microgrid, which consists of a wind turbine and solar panels. Energy storage systems support the stable and dependable functioning of the power system since the solar panel and wind turbine only occasionally produce electricity. Through various case studies, the proposed plan is tested and found to be effective in reducing electricity costs and the peak to average ratio while maximizing profit. Furthermore, a comparative study is conducted to demonstrate the legality and effectiveness of LMMRFO and MRFO.


Slow cell infection, inefficient primary infection and inability to replicate in the fat body determine the host range of Thysanoplusia orichalcea nucleopolyhedrovirus.
Thysanoplusia orichacea multicapsid nucleopolyhedrovirus (ThorMNPV) carrying an enhanced green fluorescent protein (EGFP) gene expression cassette (vThGFP) was used to study host-range mechanisms. Infection kinetics showed that vThGFP replication in Sf21 cells was too slow to suppress cell growth. Wide-host-range Autographa californica MNPV (AcMNPV) could speed up vThGFP infection and enhance the vThGFP infection rate in Sf21 cells. The enhancement was not due to recombination, as no recombinant virus was isolated from co-infection by plaque assay. No improvement of vThGFP infection in Sf21 was found by AcMNPV cosmid transactivation assay. However, culture medium from Sf21 cells infected with AcMNPV did enhance vThGFP replication in Sf21. Third-instar larvae of Spodoptera frugiperda, S. exigua and Helicoverpa zea were not killed by feeding with vThGFP polyhedra but were killed by intrahaemocoelic injection using budded viruses (BVs). This suggested that insufficient BVs were generated during the primary infection in the midgut. vThGFP infected haemocytes, tracheae and Malpighian tubules but not fat bodies of larvae of S. frugiperda, S. exigua and H. zea. Third-instar S. frugiperda larvae co-infected by injection with vThGFP and vAcDsRed2, an AcMNPV expressing a red fluorescent protein gene, showed EGFP expression in the fat body. This result suggests that vAcDsRed2 could help vThGFP to replicate in the fat body or trans-activate EGFP expression in the fat body. All these results suggested that slow cell infection, insufficient primary infection and inability to replicate in the fat body control the host range of ThorMNPV.


Symptoms at the time of arrhythmia recurrence in patients receiving azimilide for control of atrial fibrillation or flutter: results from randomized trials.
Azimilide is a new antiarrhythmic agent being developed for the management for atrial fibrillation and flutter (AF). Four randomized, placebo-controlled, double-blind trials have been performed that investigated the effect of azimilide on time to first recurrence of symptomatic AF. This paper examines the data collected during those studies regarding the symptoms reported by patients at the time of AF recurrence At the time that patients reported their first documented symptomatic recurrence of arrhythmia, they were systematically asked whether or not they were experiencing any of the following 6 symptoms: palpitation, fatigue, chest pain, shortness of breath, dizziness, or sweating. Patients were required to answer yes or no. A symptom score was created varying from 0 to 6, in increasing order of number of symptoms reported. This was compared for patients receiving either of 2 doses of azimilide or placebo. The relationship between the number of symptoms, heart rate at time of arrhythmia recurrence and treatment was analyzed. In 2 separate studies, azimilide at a dose of 125 mg/day significantly reduced the number of symptoms at the time of arrhythmia recurrence compared to placebo. On the other hand, in 2 studies, the dose of 100 mg/day did not significantly reduce symptom burden. The individual symptoms significantly reduced by azimilide125 mg/day were fatigue, shortness of breath, chest pain and dizziness. Palpitations and sweating were not significantly reduced. Modeling of heart rate at the time of arrhythmia recurrence, symptoms and treatment indicated that a small reduction in heart rate with azimilide accounted for only a small part of the symptom reduction. There was another effect of azimilide: an average reduction of 0.38 symptoms (P <.01) that was independent of heart rate. Azimilide (125 mg/day) reduces the number of symptoms reported at the time of AF recurrence.


Infrared imaging in breast cancer: automated tissue component recognition and spectral characterization of breast cancer cells as well as the tumor microenvironment.
Current evaluation of histological sections of breast cancer samples remains unsatisfactory. The search for new predictive and prognostic factors is ongoing. Infrared spectroscopy and its potential to probe tissues and cells at the molecular level without requirement for contrast agents could be an attractive tool for clinical and diagnostic analysis of breast cancer. In this study, we report the successful application of FTIR (Fourier transform infrared) imaging for breast tissue component characterization. We show that specific FTIR spectral signatures can be assigned to the major tissue components of breast tumor samples. We demonstrate that a tissue component classifier can be built based on a spectral database of well-annotated tissues and successfully validated on independent breast samples. We also demonstrate that spectral features can reveal subtle differences within a tissue component, capturing for instance lymphocytic and stromal activation. By investigating in parallel lymph nodes, tonsils and wound healing tissues, we prove the uniqueness of the signature of both lymphocytic infiltrate and tumor microenvironment in the breast disease context. Finally, we demonstrate that the biochemical information reflected in the epithelial spectra might be clinically relevant for the grading purpose, suggesting potential to improve breast cancer management in the future.


Efficiency of box-traps and leg-hold traps with several bait types for capturing small carnivores (mammalia) in a disturbed area of southeastern Brazil.
Capturing small carnivores is often necessary for obtaining key ecological data. We compared the efficiency of box and leg-hold traps, using live and dead bait, to capture six carnivore species (Herpailurus yagouaroundi (E. Geoffroyi, 1803), Leopardus tigrinus (Schreber, 1775), Nasua nasua (Linnaeus, 1766), Cerdocyon thous (Linnaeus, 1766), Eira barbara (Linnaeus, 1758), and Galictis cuja (Molina, 1782)). The use of leg-hold traps significantly increased the capture rate of carnivores (5.77%) and non-target species (non-carnivores, 11.54%). Dead bait significantly attracted more non-carnivores than carnivores and live bait was more efficient for capturing carnivores (2.56%) than non-carnivores (0.77%). Both box and leg-hold traps caused some minor injuries (swelling and claw loss). We provide recommendations for the ethical use of these trap and bait types.


Nursing theory views on how to support the process of ageing.
An important purpose of theories is to challenge practice, create new approaches to practice and remodel the structures of rules and principles. Therefore, it is of interest to study whether it is possible to make use of nursing theories in care of older people, when the aim is to support them in the process of ageing. To review 17 well-known nursing theories/models in order to delineate the views on ageing presented and discover whether there are descriptions of how nursing care of older people could be organized. Seventeen nursing theories/models originating over a 30-year period from the early 1960s to the early 1990s were chosen. A search of CINAHL was done and lists of reference from books were used to find original sources. For the review, a list of research questions were formulated and addressed to each theory. Results showed that most nursing theorists see human ageing from a developmental perspective, but do not discuss what ageing implies. In only a few theories are some important aspects of nursing care of older people discussed, but no concrete instructions are given on how to apply these to nursing care. There is a need to develop a nursing care model that - more than other contemporary theories - takes into consideration human ageing.


Effect of acid etching and different Er:YAG laser procedures on microleakage of three different fissure sealants in primary teeth after aging.
The purpose of this study was to evaluate the effects of different surface conditioning procedures on microleakage and unfilled area proportions of three fissure sealants(Aegis, Helioseal and Helioseal F) in primary molar teeth. One hundred and fifty teeth were randomly divided into five main groups according to surface conditioning procedures. Main groups were; acid-etching(Group A); laser 3.75 W(Group B) and 5 W(Group D); acid-etching combined with laser 3.75 W(Group C) and 5 W(Group E). Samples were aged by thermocycling and water bath. Afterwards, they were immersed in basic fuchsin solution for 24 h and bucco-lingual sections were obtained from each tooth. Digital images were taken using a stereo-microscope and the microleakage and unfilled area proportions were assessed by using a software system. Consequently, Aegis FS containing amorphous calcium phosphate was found to exhibit the lowest microleakage in all surface conditioning procedures. Unfilled areas were not detected in Groups C and E of all FSs.


Effects of tableting process parameters and powder lubrication levels on tablet surface temperature and moisture content.
Punch sticking is a recurrent problem during the pharmaceutical tableting process. Powder moisture content plays a key role in the buildup of sticking; it evaporates due to increased tablet temperature, accumulates at the punch-tablet interface, and causes sticking through capillary force. This study investigated the effects of compaction pressure (CP), compaction speed (CS), and lubrication level (magnesium stearate (MgSt) ratio) on tablet surface temperature (TST) and tablet surface moisture content (TSMC). TST and TSMC were measured with an infrared thermal camera and near-infrared sensor, respectively. Microcrystalline cellulose was used as the tableting powder and MgSt as the lubricant. The low range of CS values (16-32 mm/s) considered in this study did not have significant effects on TST and TSMC. MgSt ratio had a significant positive effect on TST; this may be explained by the increase in powder blend effusivity with the addition of MgSt. However, MgSt ratio did not have a significant effect on TSMC. CP had a significant positive effect on both TST and TSMC. Increased CP induced higher heat generation through particle deformation and friction during the compaction phase, leading to increased TST. Furthermore, the water vapor diffusion rate through the powder bed might have increased due to the rise in thermal energy and led to further moisture accumulation at the tablet-punch interface, causing the significant positive effect of CP on TSMC. This result may explain the occurrence of sticking regardless of the CP applied during the tableting process.


Purification and characterization of antifungal δ-dodecalactone from Lactobacillus plantarum AF1 isolated from kimchi.
The aim of this study was to purify and identify an antifungal compound from Lactobacillus plantarum AF1, which was isolated from kimchi. The antifungal compound was purified by solid-phase extraction and recycling preparative high-performance liquid chromatography, and its structure was elucidated by using gas chromatography-mass spectrometry (GC-MS). The active compound from L. plantarum AF1 was confirmed to be δ-dodecalactone (molecular weight, 198.3) by comparison of its gas chromatographic retention time with the mass spectrum of standard δ-dodecalactone. The MICs of δ-dodecalactone against various fungi and bacteria ranged from 350 to 6,250 m g/ml. δ-Dodecalactone showed strong antifungal activity against molds Aspergillus flavus, A. fumigatus, A. petrakii, A. ochraceus, A. nidulans, and Penicillium roqueforti. The three tested yeast strains of Candida albicans were more resistant than the molds. Antibacterial activity was evident but less potent than the antifungal activity. δ-Dodecalactone produced pleasurable (fruity) organoleptic characteristics. The results indicate the potential of the δ-dodecalactone produced by L. plantarum AF1 as a biopreservative and flavoring compound, as well as a biosafe remedy for candidiasis.


DNA methylation and expression of the folate transporter genes in colorectal cancer.
Folate has a central role in the cell metabolism. This study aims to explore the DNA methylation pattern of the folate transporter genes FOLR1, PCFT, and RFC1 as well as the corresponding protein expressions in colorectal cancer (CRC) tissue and adjacent non-cancerous mucosa (ANCM). Our results showed statistically significant differences in the DNA-methylated fraction of all three genes at several gene regions; we identified three differentially methylated CpG sites in the FOLR1 gene, five CpG sites in the PCFT gene, and six CpG sites in the RFC1 gene. There was a pronounced expression of the FRα and RFC proteins in both the CRC and ANCM tissues, though the expression was attenuated in cancer compared to the paired ANCM tissues. The PCFT protein was undetectable or expressed at a very low level in both tissue types. Higher methylated fractions of the CpG sites 3-5 in the RFC1 gene were associated with a lower protein expression, suggestive of epigenetic regulation by DNA methylation of the RFC1 gene in the colorectal cancer. Our results did not show any association between the RFC and FRα protein expression and tumor stage, TNM classification, or tumor location. In conclusion, this is the first study to simultaneously evaluate both DNA methylation and protein expression of all three folate transporter genes, FOLR1, PCFT, and RFC1, in colorectal cancer. The results encourage further investigation into the possible prognostic implications of folate transporter expression and DNA methylation.


Perceived Satisfaction With Long-Term Oxygen Delivery Devices Affects Perceived Mobility and Quality of Life of Oxygen-Dependent Individuals With COPD.
Although routine physical activity for individuals with COPD is recommended, there are inherent limitations of available oxygen delivery devices that may result in hypoxemia during activity. Changes in Medicare laws have resulted in an increased use of oxygen cylinders and a reduction in the use of liquid oxygen devices. The aim of this survey was to assess the impact of perceived satisfaction with various oxygen delivery devices on perceived mobility and quality of life (QOL) of oxygen-dependent subjects with COPD. A survey was developed to measure perceived satisfaction with current portable oxygen delivery devices, perceived mobility, and perceived QOL. The survey was deployed via a link posted on the COPD Foundation's COPD360SOCIAL social media site for 5 weeks, which resulted in the recruitment of 529 participants, of which 417 were included in the data analysis. Quantile regression analysis revealed that the median perceived device satisfaction score was significantly higher in the liquid oxygen device group (P < .001) compared with the portable oxygen cylinder and portable oxygen concentrator (POC) groups. The median perceived mobility score was significantly higher in the liquid oxygen device group (P < .001) compared with the portable oxygen cylinder group, but not the POC group. The median QOL score was significantly higher in the liquid oxygen device group (P < .001) compared with the POC and portable oxygen cylinder groups. Moreover, partial least-squares structural equation modeling regression analysis showed that perceived mobility is significantly affected by perceived satisfaction with the long-term oxygen therapy (LTOT) device (adjusted R2 = 0.15, P < .001), and perceived QOL is significantly affected by both perceived satisfaction with the LTOT device and perceived mobility (adjusted R2 = 0.45, P < .001). For individuals with COPD requiring LTOT, perceived satisfaction with a portable LTOT device significantly and positively affects perceived mobility and QOL.


CpG-ODN-induced nitric oxide production is mediated through clathrin-dependent endocytosis, endosomal maturation, and activation of PKC, MEK1/2 and p38 MAPK, and NF-kappaB pathways in avian macrophage cells (HD11).
We have characterized the nitric oxide (NO) induction by CpG oligodeoxydinucleotide (CpG-ODN) and lipopolysaccharide (LPS) in an avian macrophage cell line (HD11) and evaluated signal transduction pathways by using selective inhibitors. Our results indicate that while CpG-ODN and LPS both stimulate inducible NO synthase (iNOS) to produce NO through common signalling pathways involving activation of protein kinase C (PKC), mitogen-activated protein kinases (p38 MAPK and MEK1/2) and transcription factor NF-kappaB; CpG-ODN inducing NO production distinctively requires a clathrin-dependent endocytosis and subsequent endosomal maturation. Inhibitors of clathrin-dependent endocytosis such as monodansylcadaverine and hyperosmolar sucrose completely abolished CpG-ODN stimulated NO production by HD11 cells, but have no or less effect on LPS-induced NO production. The endosomal maturation is also critical for stimulation of NO induction by CpG-ODN, but not by LPS. Our findings are the first to demonstrate cellular signalling pathways that mediate CpG-ODN immunostimulatory activity in cells from non-mammalian species.


Clinical implications of gene discovery in Parkinson's disease and parkinsonism.
Over the past decade, major progress has been achieved in the identification of genes associated with Parkinson's disease (PD) and parkinsonism. Five genes have now been shown conclusively to play a role in PD susceptibility. Mutations in three of these genes, PRKN, PINK1, and DJ1, are important in early onset, recessively inherited PD, while mutations in LRRK2 and SNCA result in autosomal-dominant PD. LRRK2 has emerged as the most prevalent genetic cause of PD and has been implicated in both familial and sporadic forms of disease. In addition, autosomal-dominant dementia and Parkinsonism has been shown to be caused by mutations in the MAPT and PGRN genes. Molecular tests are now commercially available for several of these genes; however, in some of them, positive results need to be interpreted with caution until penetrance is better understood. In addition, clinical treatment of PD remains largely unaltered by the results of genetic testing.


Aberrant distributions of nuclear pore complex proteins in ALS mice and ALS patients.
Nuclear pore complexes (NPCs) play important roles in traffic of molecules between the nucleus and cytoplasm, aberrant distributions of components of NPCs were demonstrated in C9orf72 amyotrophic lateral sclerosis (C9-ALS) patients, but it is elusive whether such abnormities are also the case with other cause of ALS disease. In the present study, we investigated the spatiotemporal distributions of RanGAP1 and 4 representative nucleoporins (GP210, NUP205, NUP107 and NUP50) of NPCs in human Cu/Zn superoxide dismutase-1 mutation transgenic (SOD1-Tg) mice and sporadic ALS patients. Compared with wild type (WT), these proteins displayed age-dependent and progressive nuclear precipitations, and cytoplasmic aberrant expressions in motor neurons of lumbar cord in SOD1-Tg mice from 10 to 18weeks (W). Double immunofluorescent analysis showed abnormal nuclear retention and apparent co-localizations of RanGAPl with NUP205 and NUP205 with NUPl07, meanwhile, GP210 with NUP205 mainly co-localized in the nuclear envelope (NE) of motor neurons. Furthermore, RanGAP1, GP210 and NUP50 showed similarly abnormal nuclear precipitations and cytoplasmic upregulations in SOD1-Tg mice and ALS patients, moreover, aberrant co-localizations of RanGAP1 with TDP-43 and NUP205 with TDP-43 were also observed in motor neurons. The present study indicated that the mislocalization of these proteins of NPCs may underlie the pathogenesis of ALS both in SOD1-Tg mice and human sporadic ALS patients, and these dysfunctions may be a fundamental pathway for ALS that is not specific only in C9-ALS but also in SOD1-ALS, which may be amenable to pharmacotherapeutic intervention.


Development and residue screening of the furazolidone metabolite, 3-amino-2-oxazolidinone (AOZ), in cultured fish by an enzyme-linked immunosorbent assay.
A sensitive and specific polyclonal enzyme-linked immunosorbent assay (ELISA) for the determination of tissue-bound metabolite 3-amino-2-oxazolidinone (AOZ) is described. The procedures allow for the detection of protein-bound AOZ in the form of a 2-nitrophenyl derivative (2-NP-AOZ) in the sample supernatant or extract after acid hydrolysis and derivatization with o-nitrobenzaldehyde. The polyclonal rabbit antibodies were produced with the immunogen hapten, 2-NP-HXA-AOZ, and the 50% inhibition values (IC(50)) of 0.14 microg kg(-1) of AOZ was achieved with the most sensitive antibody A0505. The mean lower detection limit of the ELISA method is about 0.025 microg kg(-1). According to the test preparation record, the detection limit is 0.1 microg kg(-1), which is well below the minimum required performance limits (MRPLs) for tissue-bound residues of AOZ at 1 microg kg(-1) in the European Communities. In the present study, we investigated the use of homemade ELISA, a new immunoassay, to monitor the presence of the furazolidone marker residue in 370 samples of cultured fish. Adopting 0.3 microg kg(-1) AOZ as a cutoff value, the ELISA has a sensitivity of 100% and a specificity of 98.5% versus high-performance liquid chromatography-mass spectrometry (HPLC-MS) at a cutoff of 0.3 microg kg(-1) and gives no false-negative rate results. From the practical point of view, the homemade kit could be advantageously used for the screening of large groups of animal-edible tissue samples and the kit employed has good reliability even in routine application for the control of the illegal use of the drug.


Cutaneous group B streptococcal infection.
A 20-year-old man had a penile ulceration from which a pure, heavy growth of group B streptococcus was cultured. Cutaneous infection with this organism is unusual, and it is most often reported in postpartum women, diabetic patients, and immunocompromised individuals. Vaginal colonization with group B streptococcus is common. Since traumatic erosions or ulcerations of the penis are not uncommon, it is likely that a certain number of unrecognized infections of this type occur.


Comparative morphology of the euglenid pellicle. II. Diversity of strip substructure.
The morphological diversity associated with the strip substructure of the euglenid pellicle was examined, and after identifying characters and states, we outlined hypotheses about their evolution. We have attempted to standardize terms necessary for analytical comparisons of strips by providing a glossary and comparing published synonyms. Most of the substructural diversity found in euglenids is demonstrated with 13 representative taxa. Strips are generally composed of two subcomponents: frames and projections. Frames support the basic shape of strips and many can be described as either S-shaped, plateau-shaped, M-shaped, or A-shaped. Projections branch laterally from the frames, are usually periodic, and can be described as thread-like structures, an indented plate, tooth-like structures, and plate-like structures. The ancestral state included strips that were few in number, flat, and fused. The strips became S-shaped and disjoined in the lineage leading to most euglenid taxa. These strips became secondarily flattened and fused in one lineage. In some lineages of phototrophs, the strips became increasingly robust. Two strips of different morphology formed the repeating pellicular unit or doublet in four taxa. These doublets evolved convergently at least three times and may provide insights into developmental patterns of the cytoskeleton.


The development of the head direction system before eye opening in the rat.
Head direction (HD) cells are neurons found in the hippocampal formation and connected areas that fire as a function of an animal's directional orientation relative to its environment. They integrate self-motion and environmental sensory information to update directional heading. Visual landmarks, in particular, exert strong control over the preferred direction of HD cell firing. The HD signal has previously been shown to appear adult-like as early as postnatal day 16 (P16) in the rat pup, just after eye opening and coinciding with the first spontaneous exploration of its environment. In order to determine whether the HD circuit can begin its organization prior to the onset of patterned vision, we recorded from the anterodorsal thalamic nucleus (ADN) and its postsynaptic target in the hippocampal formation, the dorsal pre-subiculum (PrSd), before and after eye opening in pre-weanling rats. We find that HD cells can be recorded at the earliest age sampled (P12), several days before eye opening. However, this early HD signal displays low directional information content and lacks stability both within and across trials. Following eye opening, the HD system matures rapidly, as more cells exhibit directional firing, and the quality and reliability of the directional signal improves dramatically. Cue-rotation experiments show that a prominent visual landmark is able to control HD responses within 24 hr of eye opening. Together, the results suggest that the directional network can be organized independently of visual spatial information while demonstrating the importance of patterned vision for accurate and reliable orientation in space.


[Home care for the elderly: problem or solution?].
Home care for the elderly has become an important health care tool in both developed and developing countries. However, several ethical, social, and operational concerns have received insufficient attention, and the Brazilian literature on this theme is limited. Starting with a bibliographic review on home care, this paper takes a bioethical approach to potential problems arising from this growing and important patient care modality. A broader ethical approach is needed to implement home care for the elderly, with policies to protect the patient, family, and caregiver, aimed at improving the quality of this program format.


Comprehensive Protein Interactome Analysis of a Key RNA Helicase: Detection of Novel Stress Granule Proteins.
DDX6 (p54/RCK) is a human RNA helicase with central roles in mRNA decay and translation repression. To help our understanding of how DDX6 performs these multiple functions, we conducted the first unbiased, large-scale study to map the DDX6-centric protein-protein interactome using immunoprecipitation and mass spectrometry. Using DDX6 as bait, we identify a high-confidence and high-quality set of protein interaction partners which are enriched for functions in RNA metabolism and ribosomal proteins. The screen is highly specific, maximizing the number of true positives, as demonstrated by the validation of 81% (47/58) of the RNA-independent interactors through known functions and interactions. Importantly, we minimize the number of indirect interaction partners through use of a nuclease-based digestion to eliminate RNA. We describe eleven new interactors, including proteins involved in splicing which is an as-yet unknown role for DDX6. We validated and characterized in more detail the interaction of DDX6 with Nuclear fragile X mental retardation-interacting protein 2 (NUFIP2) and with two previously uncharacterized proteins, FAM195A and FAM195B (here referred to as granulin-1 and granulin-2, or GRAN1 and GRAN2). We show that NUFIP2, GRAN1, and GRAN2 are not P-body components, but re-localize to stress granules upon exposure to stress, suggesting a function in translation repression in the cellular stress response. Using a complementary analysis that resolved DDX6's multiple complex memberships, we further validated these interaction partners and the presence of splicing factors. As DDX6 also interacts with the E3 SUMO ligase TIF1β, we tested for and observed a significant enrichment of sumoylation amongst DDX6's interaction partners. Our results represent the most comprehensive screen for direct interaction partners of a key regulator of RNA life cycle and localization, highlighting new stress granule components and possible DDX6 functions-many of which are likely conserved across eukaryotes.


The role of prostacyclin in modifying acute hepatotoxicity of acetaminophen in mice.
Prostaglandins (PGs) are lipid compounds that mediate the variety of physiological and pathological functions in almost all body tissues and organs. Prostacyclin (prostaglandin 12, PGI2), which is synthesized by the vascular endothelium, is a potent vasodilator, inhibits the aggregation of platelets in vitro and has cytoprotective effect on gastrointestinal mucosa. The aim of this study was to determine whether PGI2 is playing a role in host defense to toxic effect of acetaminophen (APAP). This was investigated in C57Black/6 mice which were intoxicated with single lethal or high sublethal dose of APAP. APAP was administered to mice by gastric lavage and PGI2 agonists or antagonists were given intraperitoneally (i.p.) 30 minutes before or 2 hours after administration of APAP. The toxicity of APAP was determined by observing the survival of mice during 48 hours, by measuring the concentration of alanine-aminotransferase (ALT) in plasma 20-24 hours after APAP administration, and by liver histology. Mice were given either pure PGI2 (PGI2 sodium salt), its stable agonist (iloprost) or inhibitor of prostacyclin (IP)-receptor (CAY-10441). The results have shown that PGI2 exibits a strong hepatoprotective effect when it was given to mice either before or after APAP (both increase of survival of mice and decrease of plasma ALT levels were statistical significant). Iloprost has not shown a similar effect and CAY-10441 increased toxic effect of APAP if given 2 hours after its administration. Histopathological changes in liver generally support these findings. These investigations support the view that PGI2 is involved in defense of organism to noxious effects of xenobiotics on liver.


Mortality risk for patients with cytomegalovirus retinitis and acquired immune deficiency syndrome.
We prospectively followed up 589 patients to evaluate the relationship of anti-cytomegalovirus (CMV) treatment and immune reconstitution in response to highly active antiretroviral therapy (HAART) on the mortality risk of patients with CMV retinitis and acquired immune deficiency syndrome. The use of HAART was associated with an 81% lower mortality rate (95% confidence interval [CI], 74%-86%); it was 96% lower (95% CI, 92%-98%) for those who developed immune recovery and 49% lower (95% CI, 30-63%) for those who did not. Using time-updated multivariate analysis, current systemic anti-CMV treatment was independently associated with a 28% lower mortality rate (95% CI, 8%-43%). On the basis of these results, for patients who continue to have profound immunodeficiency despite HAART, the continued use of HAART and systemic anti-CMV therapy is predicted to reduce the risk of mortality by 65%, over and above the benefits of Pneumocystis carinii and Mycobacterium avium prophylaxis.


The Role of the Cerebellum in Learning to Predict Reward: Evidence from Cerebellar Ataxia.
Recent findings in animals have challenged the traditional view of the cerebellum solely as the site of motor control, suggesting that the cerebellum may also be important for learning to predict reward from trial-and-error feedback. Yet, evidence for the role of the cerebellum in reward learning in humans is lacking. Moreover, open questions remain about which specific aspects of reward learning the cerebellum may contribute to. Here we address this gap through an investigation of multiple forms of reward learning in individuals with cerebellum dysfunction, represented by cerebellar ataxia cases. Nineteen participants with cerebellar ataxia and 57 age- and sex-matched healthy controls completed two separate tasks that required learning about reward contingencies from trial-and-error. To probe the selectivity of reward learning processes, the tasks differed in their underlying structure: while one task measured incremental reward learning ability alone, the other allowed participants to use an alternative learning strategy based on episodic memory alongside incremental reward learning. We found that individuals with cerebellar ataxia were profoundly impaired at reward learning from trial-and-error feedback on both tasks, but retained the ability to learn to predict reward based on episodic memory. These findings provide evidence from humans for a specific and necessary role for the cerebellum in incremental learning of reward associations based on reinforcement. More broadly, the findings suggest that alongside its role in motor learning, the cerebellum likely operates in concert with the basal ganglia to support reinforcement learning from reward.


Cyclic fatigue resistance of two reciprocating nickel-titanium instruments after immersion in sodium hypochlorite.
To assess resistance to cyclic fatigue of reciprocating nickel-titanium ( NiTi ) files (Reciproc and WaveOne) after immersion in NaOCl solution over several time periods. A total of 90 new Reciproc R25 and WaveOne Primary were tested. The 45 files of the same brand were randomly assigned to three groups (n = 15) and submitted to the following immersion protocol in 5% NaOCl at 37 °C for 16 mm: no immersion (control), 1 or 5 min dynamically. The dynamic immersion and the following cyclic fatigue tests were performed using the appropriate preset reciprocation modes ('RECIPROC ALL' or 'WAVEONE ALL') in a specially designed endodontic motor. Resistance to cyclic fatigue was determined by recording time to fracture (TtF) in a stainless steel artificial canal with a 60° angle of curvature and 5 mm radius of curvature. The artificial canal was manufactured reproducing the instrument's size and taper. Data were analysed by 2-way analyses of variance. Resistance to cyclic fatigue of the same NiTi file was not significantly affected by immersion in NaOCl . Reciproc R25 was associated with a higher cyclic fatigue resistance in all groups compared to WaveOne Primary (P < 0.0001). Reciprocating dynamic immersion in NaOCl for 1 or 5 min did not reduce the cyclic fatigue resistance of NiTi files significantly. However, the type of reciprocating instrument influenced cyclic fatigue resistance with Reciproc R25 being more resistant than WaveOne Primary.


A comparative simulation study of AR(1) estimators in short time series.
Various estimators of the autoregressive model exist. We compare their performance in estimating the autocorrelation in short time series. In Study 1, under correct model specification, we compare the frequentist r1 estimator, C-statistic, ordinary least squares estimator (OLS) and maximum likelihood estimator (MLE), and a Bayesian method, considering flat (Bf) and symmetrized reference (Bsr) priors. In a completely crossed experimental design we vary lengths of time series (i.e., T = 10, 25, 40, 50 and 100) and autocorrelation (from -0.90 to 0.90 with steps of 0.10). The results show a lowest bias for the Bsr, and a lowest variability for r1. The power in different conditions is highest for Bsr and OLS. For T = 10, the absolute performance of all measurements is poor, as expected. In Study 2, we study robustness of the methods through misspecification by generating the data according to an ARMA(1,1) model, but still analysing the data with an AR(1) model. We use the two methods with the lowest bias for this study, i.e., Bsr and MLE. The bias gets larger when the non-modelled moving average parameter becomes larger. Both the variability and power show dependency on the non-modelled parameter. The differences between the two estimation methods are negligible for all measurements.


CD4+ T cells from human neonates and infants are poised spontaneously to run a nonclassical IL-4 program.
Senescence or biological aging impacts a vast variety of molecular and cellular processes. To date, it is unknown whether CD4(+) Th cells display an age-dependent bias for development into specific subpopulations. In this study, we show the appearance of a distinct CD4(+) T cell subset expressing IL-4 at an early stage of development in infant adenoids and cord blood that is lost during aging. We identified by flow cytometric, fluorescent microscopic, immunoblot, and mass spectrometric analysis a population of CD4(+) T cells that expressed an unglycosylated isoform of IL-4. This T cell subpopulation was found in neonatal but not in adult CD4(+) T cells. Furthermore, we show that the mRNA of the Th2 master transcription factor GATA3 is preferentially expressed in neonatal CD4(+) T cells. The Th2 phenotype of the IL-4(+)CD4(+) T cells could be reinforced in the presence of TGF-β. Although the IL-4(+)CD4(+) T cells most likely originate from CD31(+)CD4(+) T recent thymic emigrants, CD31 was downregulated prior to secretion of IL-4. Notably, the secretion of IL-4 requires a so far unidentified trigger in neonatal T cells. This emphasizes that cytokine expression and secretion are differentially regulated processes. Our data support the hypothesis of an endogenously poised cytokine profile in neonates and suggest a link between cytokine production and the developmental stage of an organism. The determination of the IL-4 isoform-expressing cells in humans might allow the identification of Th2 precursor cells, which could provide novel intervention strategies directed against Th2-driven immunopathologies such as allergies.