Familial ALS-associated SFPQ variants promote the formation of SFPQ cytoplasmic aggregates in primary neurons.
Splicing factor proline- and glutamine-rich (SFPQ) is a nuclear RNA-binding protein that is involved in a wide range of physiological processes including neuronal development and homeostasis. However, the mislocalization and cytoplasmic aggregation of SFPQ are associated with the pathophysiology of amyotrophic lateral sclerosis (ALS). We have previously reported that zinc mediates SFPQ polymerization and promotes the formation of cytoplasmic aggregates in neurons. Here we characterize two familial ALS (fALS)-associated SFPQ variants, which cause amino acid substitutions in the proximity of the SFPQ zinc-coordinating centre (N533H and L534I). Both mutants display increased zinc-binding affinities, which can be explained by the presence of a second zinc-binding site revealed by the 1.83 Å crystal structure of the human SFPQ L534I mutant. Overexpression of these fALS-associated mutants significantly increases the number of SFPQ cytoplasmic aggregates in primary neurons. Although they do not affect the density of dendritic spines, the presence of SFPQ cytoplasmic aggregates causes a marked reduction in the levels of the GluA1, but not the GluA2 subunit of AMPA-type glutamate receptors on the neuronal surface. Taken together, our data demonstrate that fALS-associated mutations enhance the propensity of SFPQ to bind zinc and form aggregates, leading to the dysregulation of AMPA receptor subunit composition, which may contribute to neuronal dysfunction in ALS.


Evaluation of three diagnostic methods for initial proximal caries detection in primary molars.
The proximal surfaces of primary molars in 100 selected children were examined for initial caries using three different diagnostic methods: 1--Clinical examination. 2--Fiber-optic transillumination. and 3--Bite-wing radiographic examination. Clinical examination revealed 65 initial proximal lesions, 101 lesions were detected by fiber-optic transillumination, while bite-wing radiographs verified 131 lesions. Agreement between clinical and bite-wing radiographic examinations was 49.6%, while fiber-optic transillumination showed a 77.1% agreement with bite-wing radiographic examination. The three tested diagnostic methods showed no significant difference in detecting proximal lesions in the upper and lower molars, the right and left sides or the mesial and distal surfaces, except for clinical examination which tended to miss more distal lesions.


Computation of measures of effect size for neuroscience data sets.
The overwhelming majority of research in the neurosciences employs P-values stemming from tests of statistical significance to decide on the presence or absence of an effect of some treatment variable. Although a continuous variable, the P-value is commonly used to reach a dichotomous decision about the presence of an effect around an arbitrary criterion of 0.05. This analysis strategy is widely used, but has been heavily criticized in the past decades. To counter frequent misinterpretations of P-values, it has been advocated to complement or replace P-values with measures of effect size (MES). Many psychological, biological and medical journals now recommend reporting appropriate MES. One hindrance to the more frequent use of MES may be their scarcity in standard statistical software packages. Also, the arguably most widespread data analysis software in neuroscience, matlab, does not provide MES beyond correlation and receiver-operating characteristic analysis. Here we review the most common criticisms of significance testing and provide several examples from neuroscience where use of MES conveys insights not amenable through the use of P-values alone. We introduce an open-access matlab toolbox providing a wide range of MES to complement the frequently used types of hypothesis tests, such as t-tests and analysis of variance. The accompanying documentation provides calculation formulae, intuitive explanations and example calculations for each measure. The toolbox described is usable without sophisticated statistical knowledge and should be useful to neuroscientists wishing to enhance their repertoire of statistical reporting.


Bilateral breast cancer in northern Alberta: risk factors and survival patterns.
Of 2231 women with stage I, II or III breast cancer who were registered and seen between 1971 and 1979 and followed to the end of 1981, 48 (2.2%) had synchronous and 58 (2.6%) asynchronous bilateral breast cancer. The unadjusted incidence rate for a second breast cancer was 6.4/1000 breast-years at risk, compared with a rate of 0.70 for the risk of a first breast cancer in women. When calculated from the date of diagnosis of the first breast cancer the survival rate was better for the group with asynchronous disease than for the group with synchronous disease or for a group with unilateral disease, but when calculated from the date of diagnosis of the second cancer the rate was the same in all three groups. Comparison of known risk factors showed a significant association between the development of bilateral cancer and a later age at the birth of the first child and a longer interval between menarche and that birth. There was a trend towards greater age and more stage III cancer in the group with synchronous disease. There was no correlation between receiving radiotherapy for the first breast cancer and development of the second cancer. Annual mammography and clinical examination of asymptomatic women at a cancer centre resulted in the detection of a significantly higher proportion of minimal breast cancers in the second breast compared with the first. Such screening practices should be even more valuable in the earlier detection of unilateral breast cancer in asymptomatic women who have not had breast cancer.


Concordance of p16INK4a and E6*I mRNA among HPV-DNA-Positive Oropharyngeal, Laryngeal, and Oral Cavity Carcinomas from the ICO International Study.
Background: Tests or test algorithms for diagnosing HPV-driven oral cavity and laryngeal head and neck carcinomas (HNC) have not been yet validated, and the differences among oral cavity and laryngeal sites have not been comprehensively evaluated. We aimed to assess the utility of a diagnostic algorithm for the detection of HPV-driven oral cavity (OCC), oropharyngeal (OPC) and laryngeal (LC) carcinomas using HPV-DNA testing followed by p16INK4a immunohistochemistry, taking E6*I mRNA detection as the reference standard. Methods: Formalin-fixed paraffin-embedded OCC, OPC, and LC carcinomas were collected from pathology archives in 29 countries. All samples were subjected to histopathological evaluation, DNA quality control, and HPV-DNA detection. All HPV-DNA-positive samples (including 78 OCC, 257 OPC, and 51 LC out of 3680 HNC with valid HPV-DNA results) were also tested for p16INK4a immunohistochemistry and E6*I mRNA. Three different cutoffs of nuclear and cytoplasmic staining were evaluated for p16INK4a: (a) >25%, (b) >50%, and (c) ≥70%. The concordance of p16INK4a and E6*I mRNA among HPV-DNA-positive OCC, OPC, and LC cases was assessed. Results: A total of 78 OCC, 257 OPC, and 51 LC were HPV-DNA-positive and further tested for p16INK4a and E6*I mRNA. The percentage of concordance between p16INK4a (cutoff ≥ 70%) and E6*I mRNA among HPV-DNA-positive OCC, OPC, and LC cases was 79.5% (95% CI 69.9−89.1%), 82.1% (95% CI 77.2−87.0%), and 56.9% (95% CI 42.3−71.4%), respectively. A p16INK4a cutoff of >50% improved the concordance although the improvement was not statistically significant. For most anatomical locations and p16INK4a cutoffs, the percentage of discordant cases was higher for HPV16- than HPV-non16-positive cases. Conclusions: The diagnostic algorithm of HPV-DNA testing followed by p16INK4a immunohistochemistry might be helpful in the diagnosis of HPV-driven OCC and OPC, but not LC. A different p16INK4a expression pattern was observed in those cases HPV-DNA-positive for types other than HPV16, as compared to HPV16-positive cases. Our study provides new insights into the use HPV-DNA, p16INK4a, and HPV-E6*I mRNA for diagnosing an HPV-driven HNC, including the optimal HPV test or p16INK4a cutoffs to be used. More studies are warranted to clarify the role of p16INK4a and HPV status in both OPC and non-OPC HNC.


ISSFAL 2014 Debate: It Is Time to Update Saturated Fat Recommendations.
This paper summarizes a debate on whether to update recommendations for the consumption of saturated fatty acids (SFA); this debate was held at the 11th congress of the International Society for the Study of Fatty Acids and Lipids in Stockholm, Sweden, June 28-July 2, 2014. Recommendations to reduce SFA intakes are based largely on the premise that high intakes of SFA raise low-density lipoprotein (LDL)-cholesterol levels, which in turn increase the risk of coronary heart disease (CHD). Several systematic reviews question whether reducing SFA intakes lowers CHD risk. Arguing to revise SFA recommendations, Philippe Legrand noted that SFA are heterogeneous in structure and function, are synthesized de novo by humans and only certain SFA in excess have been linked to CHD risk. We cannot consider all SFA as a block. The effects of reducing SFA intakes depend on which nutrients replace them and on which biomarkers or endpoints are assessed, Ronald Mensink observed. The effects of reducing SFA on CHD risk vary with the nutrient of comparison, whether carbohydrates, monounsaturated or polyunsaturated fatty acids. Substitution of SFA with polyunsaturated fatty acids was associated with a lower incidence of cardiovascular disease, while the effects of substitution with monounsaturated fatty acids or high-glycemic index carbohydrates are less clear.


The value of MR neurography for evaluating extraspinal neuropathic leg pain: a pictorial essay.
Fifteen patients with neuropathic leg pain referable to the lumbosacral plexus or sciatic nerve underwent high-resolution MR neurography. Thirteen of the patients also underwent routine MR imaging of the lumbar segments of the spinal cord before undergoing MR neurography. Using phased-array surface coils, we performed MR neurography with T1-weighted spin-echo and fat-saturated T2-weighted fast spin-echo or fast spin-echo inversion recovery sequences, which included coronal, oblique sagittal, and/or axial views. The lumbosacral plexus and/or sciatic nerve were identified using anatomic location, fascicular morphology, and signal intensity as discriminatory criteria. None of the routine MR imaging studies of the lumbar segments of the spinal cord established the cause of the reported symptoms. Conversely, MR neurography showed a causal abnormality accounting for the clinical findings in all 15 cases. Detected anatomic abnormalities included fibrous entrapment, muscular entrapment, vascular compression, posttraumatic injury, ischemic neuropathy, neoplastic infiltration, granulomatous infiltration, neural sheath tumor, postradiation scar tissue, and hypertrophic neuropathy.


Ber-EP4: new monoclonal antibody which distinguishes epithelia from mesothelial.
A new monoclonal antibody, Ber-EP4, directed against a partially formol resistant epitope on the protein moiety of two 34 kilodalton and 39 kilodalton glycopolypeptides on human epithelial cells is described. Immunostaining of a wide range of normal and neoplastic human tissues and cell lines showed that all carcinomas and all non-neoplastic epithelial cells, except hepatocytes, parietal cells, and apical cell layers in squamous epithelia, homogeneously expressed Ber-EP4 antigen. As Ber-EP4 does not detect any normal or neoplastic non-epithelial cells, this antibody might prove valuable for the differentiation of the following (i) non-epithelial tumours from undifferentiated carcinomas; (ii) hepatocytes from bile duct cells in certain liver diseases; (iii) mesothelial cells from carcinoma cells in lung biopsy specimens; and (iv) reactive mesothelial cells from carcinoma cells in smears of serous effusions.


Laryngeal Microsurgery for the Treatment of Vocal Nodules and Cysts in Dysphonic Children.
Vocal nodules and cysts are frequent causes of infantile dysphonia. Vocal therapy is the first treatment. Microsurgery has restricted indications, especially for nodules. To describe our experience with microsurgery for nodules and cysts in children. Dysphonic children (aged 4-18 years) with the diagnosis of nodules and vocal cysts were initially selected. Of these children, only those were included who had undergone microsurgery. For nodules and cysts, the microsurgery was indicated in cases of failure of vocal therapy and in cases of voice worsening or doubts about the diagnosis. All children were submitted to auditory perceptual vocal analysis and videolaryngostroboscopy (before and after surgery, after 6 months). Surgical outcomes were: total improvement (disappearance of vocal symptoms and of the laryngeal lesions); partial improvement (partial improvement of symptoms and/or maintenance of lesions); no improvement (maintenance or worsening of the symptoms and/or persistence of the lesions). -Results: There were 78 children with vocal nodules and 27 children with vocal cysts. Surgery was indicated for 12 children with vocal nodules (15.4%) and 12 children with cysts (44.4%). Total improvement registered for nodules and cysts was 75 and 83.4%, respectively. Partial improvement for both lesions was 25 and 16.6%, respectively. The best outcome for laryngeal microsurgery in dysphonic children was for vocal cysts. So, we encourage laryngologists for this conduct in vocal cysts. The success of microsurgery for vocal nodules was lower, and in these cases voice therapy seems to be the best treatment.


The E6 oncoproteins of high-risk papillomaviruses bind to a novel putative GAP protein, E6TP1, and target it for degradation.
The high-risk human papillomaviruses (HPVs) are associated with carcinomas of the cervix and other genital tumors. Previous studies have identified two viral oncoproteins, E6 and E7, which are expressed in the majority of HPV-associated carcinomas. The ability of high-risk HPV E6 protein to immortalize human mammary epithelial cells (MECs) has provided a single-gene model to study the mechanisms of E6-induced oncogenic transformation. In this system, the E6 protein targets the p53 tumor suppressor protein for degradation, and mutational analyses have shown that E6-induced degradation of p53 protein is required for MEC immortalization. However, the inability of most dominant-negative p53 mutants to induce efficient immortalization of MECs suggests the existence of additional targets of the HPV E6 oncoprotein. Using the yeast two-hybrid system, we have isolated a novel E6-binding protein. This polypeptide, designated E6TP1 (E6-targeted protein 1), exhibits high homology to GTPase-activating proteins for Rap, including SPA-1, tuberin, and Rap1GAP. The mRNA for E6TP1 is widely expressed in tissues and in vitro-cultured cell lines. The gene for E6TP1 localizes to chromosome 14q23.2-14q24.3 within a locus that has been shown to undergo loss of heterozygosity in malignant meningiomas. Importantly, E6TP1 is targeted for degradation by the high-risk but not the low-risk HPV E6 proteins both in vitro and in vivo. Furthermore, the immortalization-competent but not the immortalization-incompetent HPV16 E6 mutants target the E6TP1 protein for degradation. Our results identify a novel target for the E6 oncoprotein and provide a potential link between HPV E6 oncogenesis and alteration of a small G protein signaling pathway.


Compact freeform illumination system design for pattern generation with extended light sources.
One of the major problems in freeform illumination design in a geometrical optics approximation is picture generation with extended light sources. In contrast to the freeform design with zero-étendue sources, the extension of the light source leads to the typical blurring effect and a contrast reduction of the required irradiance. This effect can be minimized by increasing the distance between the freeform surface and the light source, which according to étendue conservation, results in an impractically large projection optic. To tackle this problem, we propose a design concept consisting of the combination of a pattern-generating double freeform surface for collimated beam shaping, which is calculated for a zero-étendue light source, and an imaging projection system with a telecentric object space. The design concept works independently of the shape of the emission area of the light source and does not require a representation of the extended light source by several individual wavefronts. By interpreting the pattern blurring effect as a composition of a shift contribution and a distortion contribution, we show that both can be minimized simultaneously by an appropriate placement of the object plane of the imaging optics and by making the distance between both freeform surfaces as small as possible. This allows the calculation of compact, energy-efficient freeform illumination systems for picture generation with real extended light sources. We demonstrate the significant blurring reduction by designing a simple illumination system consisting of a collimation optic, a (zero-étendue) double freeform lens for collimated beam shaping, and a projection lens for the generation of the target distribution "Elaine" with an extended Lambertian emitter of 3 mm×3 mm extension and ±42 deg maximum opening angle. For a working distance to the projection system of 500 mm and a target area of 300 mm×300 mm, a relative blurring extension of 2% is estimated, compared to 23% for a single freeform projector with the same energy throughput and similar lateral extension. The influence of the doublefreeform thickness on the blurring reduction is demonstrated, and a summary of the design procedure for the developed design concept is given.


Developmental factors affecting regeneration in the central nervous system: early but not late formed mitral cells reinnervate olfactory cortex after neonatal tract section.
If the lateral olfactory tract (LOT) of the golden hamster is transected in the first week of postnatal life, axons will grow back through the cut and reinnervate the terminal regions. Functional recovery occurs only when the terminal regions are reinnervated. The experiments reported here tested the hypothesis that reinnervation is due to neogenesis: the continued growth of newly formed axons which were not severed by the lesion. In the first experiment the birth dates of the mitral and tufted cells were determined in the hamster. It was found that mitral cells are formed on gestational days 11 and 12 (E11 and E12) and tufted cells on E11 to E14. Experiment 2 involved the combination of [3H]thymidine labeling, for the time of cell formation, with the retrograde transport of horseradish peroxidase (HRP), at a time when the LOT projections are not yet complete. The axons of early formed cells were found to reach the olfactory cortex before those of later formed cells. Experiment 3 examined the possibility that the axons which grow through an early LOT transection are new axons that had not yet reached the level of the cut. Animals were given [3H]thymidine to label the times of formation of mitral and tufted cells and then were given a transection of the LOT on postnatal day 3 (P3). After a recovery period sufficient to allow axonal regrowth and reinnervation, HRP was placed in the olfactory projection region caudal to the prior LOT section. The original hypothesis was not supported. Cells that are formed early and send out their axons early are able to reinnervate the olfactory cortex, whereas late formed cells do not. The results of this experiment suggest that the factors which prevent the regrowth of axons when the LOT is cut after P7 may depend on the stage of development of the tissue into which the axons are growing, rather than in the cells of origin and their axons.


Methyl Paraben May Increase Risk of Pruritus in African Americans Whereas Triclosan Is Inversely Associated With Pruritus and Eczema.
Phenols and parabens (P&Ps) are commonly found in skin care products. However, P&Ps' role in pruritus and eczema has not been studied. The aim of the study was to investigate the association between P&Ps, and pruritus and eczema. This is a cross-sectional population-based study of 2202 participants. We examined the association between urinary phenols (triclosan, bisphenol A, benzophenone-3) and parabens (methyl and propyl parabens) and itchy rash/eczema using the 2005-2006 National Health and Nutrition Examination Survey database. Phenols and parabens were divided into quartiles (Qs) with the first Q as the reference. We calculated odds ratios and 95% confidence intervals, adjusting for multiple variables. Urinary triclosan was inversely associated with itchy rash (P trend = 0.048). In a subpopulation analysis by race/ethnicity, urinary methyl paraben was positively associated with itchy rash in African Americans (fourth Q vs first Q: odds ratio, 12.60; 95% confidence interval, 1.03-154.06; P trend = 0.02). Triclosan was inversely associated with eczema in whites (P trend = 0.04). Methyl paraben exposure may increase the risk of itchy rash in African Americans, whereas triclosan may decrease the risk of itchy rash and eczema. The potential effect of triclosan and methyl paraben in pruritus and eczema warrants further study.


Combination Effect of Titrated Extract of Centella asiatica and Astaxanthin in a Mouse Model of Phthalic Anhydride-Induced Atopic Dermatitis.
In our previous study, we demonstrated that both titrated extract of Centella asiatica (TECA) and astaxanthin (AST) have anti-inflammatory effects in a 5% phthalic anhydride (PA) mouse model of atopic dermatitis (AD). The increasing prevalence of AD demands new therapeutic approaches for treating the disease. We investigated the therapeutic efficacy of the ointment form of TECA, AST and a TECA + AST combination in a mouse model of AD to see whether a combination of the reduced doses of 2 compounds could have a synergistic effect. An AD-like lesion was induced by the topical application of 5% PA to the dorsal ear and back skin of an Hos:HR-1 mouse. After AD induction, TECA (0.5%), AST (0.5%) and the TECA (0.25%) + AST (0.25%) combination ointment (20 μg/cm²) were spread on the dorsum of the ear or back skin 3 times a week for 4 weeks. We evaluated dermatitis severity, histopathological changes and changes in protein expression by Western blotting for inducible nitric oxide synthase (iNOS), cyclocxygenase (COX)-2, and nuclear factor (NF)-κB activity. We also measured the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and immunoglobulin E (IgE) in the blood of AD mice by enzyme-linked immunosorbent assay (ELISA). PA-induced skin morphological changes and ear thickness were significantly reduced by TECA, AST and TECA + AST treatments, but these inhibiting effects were more pronounced in the TECA + AST treatment. TECA, AST and the TECA+AST reatments inhibited the expression of iNOS and COX-2; NF-κB activity; and the release of TNF-α, IL-6 and IgE. However, the TECA+AST treatment showed additive or synergistic effects on AD. Our results demonstrate that the combination of TECA and AST could be a promising therapeutic agent for AD by inhibiting NF-κB signaling.


Appetite-Enhancing Effects of Curry Oil.
Inhalation of scent compounds with phenylpropanoidal structures, such as trans-cinnamaldehyde, is expected to increase the appetite. The scent of curry powder is well known for its appetite-enhancing effect on humans. In this work, we show that the appetite of mice after inhalation of curry powder essential oil or benzylacetone showed a similar increase. The components of curry oil, trans-cinnamaldehyde, trans-anethole, and eugenol, each showed appetite-enhancing effects; therefore, these three scent compounds may be the active compounds in curry powder oil.


Application of the Mayo primary biliary cirrhosis survival model to Mayo liver transplant patients.
Liver transplantation is considered lifesaving for selected patients with end-stage primary biliary cirrhosis (PBC). A mathematical model to predict survival in the patient with PBC who has not undergone transplantation would be valuable for improving selection of patients for and timing of transplantation and for providing control information for assessment of the efficacy of transplantation. The Cox regression method and data from 312 Mayo Clinic patients with PBC were used to develop a model based on age, total serum bilirubin, serum albumin, prothrombin time, and severity of edema. When cross-validated on an independent set of 106 Mayo patients, the model accurately predicted their survival. It was similar to two other published survival models in terms of risk measurement but had the advantage of not necessitating liver biopsy. The model was used to assess the efficacy of liver transplantation by comparing the Kaplan-Meier survival of 32 Mayo patients after transplantation with the average model prediction of survival without transplantation. Beyond 3 months after transplantation, Kaplan-Meier survival probabilities were significantly greater than control survival predicted by the model (P less than 0.001). Examples of using the model for aiding in selection of patients for and timing of transplantation are provided.


Glutaminyl-tRNA synthetase.
Among the twenty aminoacyl-tRNA synthetases glutaminyl-tRNA synthetase occupies a special position: it is one of only two enzymes of this family which is not found in all organisms, being mainly absent from gram positive eubacteria, archaebacteria and organelles. The E. coli GlnRS is relatively small with 553 amino acids and a molecular mass of 64.4 kDa and functions as a monomer. The mammalian enzymes are somewhat larger and can be parts of multienzyme complexes. Crystal structures were solved of E. coli GlnRS complexed with tRNA(Gln) and ATP, of this complex containing tRNA(Gln) replaced by unmodified tRNA(Gln), and of three complexes with mutated GlnRS enzymes. The GlnRS molecule consists of four domains, the catalytic site is located in the Rossman fold, typical for class I synthetases, and the reaction mechanism follows the normal adenylate pathway. The enzyme shows many similarities with glutamyl-tRNA synthetase; a common ancestor of both molecules is well established. In the E. coli system recognition of the cognate tRNA has been studied in many details using both natural and artificial mutants of tRNA(Gln) and of the enzyme: GlnRS recognizes mainly conventional parts of the tRNA molecule, namely some bases of the anticodon loop and parts of the acceptor stem.


Human cardiac troponin I: precise identification of antigenic epitopes and prediction of secondary structure.
The presence of human cardiac troponin I (hcTnI) in serum is considered to be a highly specific biochemical marker of acute myocardial infarction. To better understand the antigenic properties of hcTnI, a set of 68 overlapping peptides covering the complete amino acid sequence of hcTnI was prepared and used in epitope mapping experiments. All 16 anti-hcTnI monoclonal antibodies tested were found to recognize a peptide epitope, indicating that recognition by anti-hcTnI monoclonal antibodies was not dependent on the tertiary structure of the protein. Furthermore, the peptide reactivity with anti-hcTnI polyclonal antibodies indicated that most of the sequence of the protein was antigenic; in particular, the N- and C-terminal extremities were found to be the strongest antigenic regions. By using accurate secondary structure prediction methods, hcTnI was found to be an all-alpha type protein, with five regions predicted as helices. Matching the results of the epitope analysis with the structural prediction led us to the view that hcTnI is not a globular protein but probably adopts an extended conformation, allowing a large part of the amino acid sequence of this molecule to be recognized by the immune system. This improved knowledge of the antigenic and structural properties of hcTnI may help in developing new antibodies and immunoassays for use in diagnosing myocardial infarction.


Antiviral Activity of an Endogenous Parvoviral Element.
Endogenous viral elements (EVEs) are genomic DNA sequences derived from viruses. Some EVEs have open reading frames (ORFs) that can express proteins with physiological roles in their host. Furthermore, some EVEs exhibit a protective role against exogenous viral infection in their host. Endogenous parvoviral elements (EPVs) are highly represented in mammalian genomes, and although some of them contain ORFs, their function is unknown. We have shown that the locus EPV-Dependo.43-ODegus, an EPV with an intact ORF, is transcribed in Octodon degus (degu). Here we examine the antiviral activity of the protein encoded in this EPV, named DeRep. DeRep was produced in bacteria and used to generate antibodies that recognize DeRep in western blots of degu tissue. To test if DeRep could protect against exogenous parvovirus, we challenged cells with the minute virus of mice (MVM), a model autonomous parvovirus. We observed that MVM protein expression, DNA damage induced by replication, viral DNA, and cytopathic effects are reduced when DeRep is expressed in cells. The results of this study demonstrate that DeRep is expressed in degu and can inhibit parvovirus replication. This is the first time that an EPV has been shown to have antiviral activity against an exogenous virus.


Robust segmentation of tubular structures in 3-D medical images by parametric object detection and tracking.
We present a novel approach to the coarse segmentation of tubular structures in three-dimensional (3-D) image data. Our algorithm, which requires only few initial values and minimal user interaction, can be used to initialize complex deformable models and is based on an extension of the randomized hough transform (RHT), a robust method for low-dimensional parametric object detection. Tubular structures are modeled as generalized cylinders. By means of a discrete Kalman filter, they are tracked through 3-D space. Our extensions to the RHT are a feature adaptive selection of the sample size, expectation-dependent weighting of the input data, and a novel 3-D parameterization for straight elliptical cylinders. Experimental results obtained for 3-D synthetic as well as for 3-D medical images demonstrate the robustness of our approach w.r.t. image noise. We present the successful segmentation of tubular anatomical structures such as the aortic arc and the spinal cord.


Intriguing Mixed Pathologic Features in a Case of Dementia With Lewy Bodies.
Neuropathologic confirmation of dementia with Lewy bodies (DLB) involves labeling cytoplasmic Lewy body inclusions for α-synuclein in cortical and subcortical neurons. The authors studied the postmortem brain of a 78-year-old man who had a diagnosis of DLB by exclusion. The patient had symptoms ascribed to DLB that included fluctuating cognitive changes in attention and executive function with progression to dementia, visual hallucinations, and parkinsonism. Sections from the olfactory bulbs and cortical and subcortical regions were stained with periodic acid-Schiff, as well as immunolabeled with antibodies specific for α-synuclein, tau protein, β-amyloid 1-42, and Chlamydia pneumoniae. Most regions demonstrated mixed neuropathologic features, and α-synuclein was notable in Lewy bodies in the amygdala and hippocampus. Periodic acid-Schiff-positive staining was noted in bodies in the amygdala and olfactory bulbs. In this case of DLB, neuropathologic inclusions were consistent with the disease diagnosis, but also with Alzheimer disease and other neurodegenerative diseases, such as polyglucosan body disease.


Audiovisual biofeedback amplifies plantarflexor adaptation during walking among children with cerebral palsy.
Biofeedback is a promising noninvasive strategy to enhance gait training among individuals with cerebral palsy (CP). Commonly, biofeedback systems are designed to guide movement correction using audio, visual, or sensorimotor (i.e., tactile or proprioceptive) cues, each of which has demonstrated measurable success in CP. However, it is currently unclear how the modality of biofeedback may influence user response which has significant implications if systems are to be consistently adopted into clinical care. In this study, we evaluated the extent to which adolescents with CP (7M/1F; 14 [12.5,15.5] years) adapted their gait patterns during treadmill walking (6 min/modality) with audiovisual (AV), sensorimotor (SM), and combined AV + SM biofeedback before and after four acclimation sessions (20 min/session) and at a two-week follow-up. Both biofeedback systems were designed to target plantarflexor activity on the more-affected limb, as these muscles are commonly impaired in CP and impact walking function. SM biofeedback was administered using a resistive ankle exoskeleton and AV biofeedback displayed soleus activity from electromyography recordings during gait. At every visit, we measured the time-course response to each biofeedback modality to understand how the rate and magnitude of gait adaptation differed between modalities and following acclimation. Participants significantly increased soleus activity from baseline using AV + SM (42.8% [15.1, 59.6]), AV (28.5% [19.2, 58.5]), and SM (10.3% [3.2, 15.2]) biofeedback, but the rate of soleus adaptation was faster using AV + SM biofeedback than either modality alone. Further, SM-only biofeedback produced small initial increases in plantarflexor activity, but these responses were transient within and across sessions (p > 0.11). Following multi-session acclimation and at the two-week follow-up, responses to AV and AV + SM biofeedback were maintained. This study demonstrated that AV biofeedback was critical to increase plantarflexor engagement during walking, but that combining AV and SM modalities further amplified the rate of gait adaptation. Beyond improving our understanding of how individuals may differentially prioritize distinct forms of afferent information, outcomes from this study may inform the design and selection of biofeedback systems for use in clinical care.


A canine model to evaluate efficacy and safety of γ-secretase inhibitors and modulators.
Gamma-secretase, a membrane bound protease which cleaves the transmembrane protein amyloid-β protein precursor (AβPP), is a therapeutic target for Alzheimer's disease. Gamma-secretase inhibitors (GSIs) and modulators (GSMs) are being investigated as potential disease-modifying agents. Preclinical in vivo models to monitor the activity on gamma-secretase are described in different species such as mouse, rat, and guinea pigs. All these models have their value in testing compounds with amyloid lowering properties, however, compound characteristics and pharmacokinetic properties, as well as other species characteristics such as limited sampling volumes of cerebrospinal fluid (CSF), recommended the use of a larger, non-rodent animal species. For this purpose, a screening model in dogs was developed for testing GSIs and GSMs. We showed that GSIs and GSMs had a dose- and time-dependent effect on Aβ(37), Aβ(38), Aβ(40), and Aβ(42) in CSF. Changes in liver function were evidenced by a transient increase in bilirubin with the GSMs and incidental increases in alanine aminotransferase for GSMs as well as GSIs. Microarray analysis of liver biopsies enabled to elucidate potential mechanisms behind the liver function changes. The relevance of the liver findings should be further evaluated in chronic pre-clinical safety studies and in humans. Based on our data, we can conclude that the dog is a very appropriate species to assess efficacy and safety of compounds which have an effect on AβPP processing such as GSMs, GSIs, and BACE-inhibitors.


Assessment of tRNAThr and tRNAGln Variants and Mitochondrial Functionality in Parkinson's Disease (PD) Patients of Tamil Nadu Population.
Parkinson's disease (PD) is speculated with genetic and environmental factors. At molecular level, the mitochondrial impact is stated to be one of the causative reasons for PD. In this study, we investigated the mitochondrial membrane potential (MMP), reactive oxygen species (ROS) and adenosine triphosphate (ATP) levels along with mitochondrial tRNA alterations among three age categories of PD. By determining the genetic and organellar functionality using molecular techniques, the ROS levels were reported to be high with decreased MMP and ATP in the late-onset age group than in other two age categories. Likewise, the tRNA significancy in tRNAThr and tRNAGln was noticed with C4335T and G15927A mutations in late-onset and early-onset PD groups respectively. Therefore, from the findings, ageing has shown a disruption in tRNA metabolism leading to critical functioning of ATP synthesis and MMP, causing oxidative stress in PD patients. These physiological outcomes show that ageing has a keen role in the divergence of mitochondrial function, thereby proving a correlation with ageing and maintenance of mitochondrial homeostasis in PD.


[Morphofunctional rearrangement of the hepatic vasculature in the pathogenesis of portal hypertension in liver cirrhosis].
The paper presents an update on the mechanisms for enhanced hepatic vascular resistance to the portal circulation underlying the pathogenesis of portal hypertension in liver cirrhosis. In addition to gross hepatic structural disorders related to diffuse fibrosis and formation of regenerative nodules, the morphofunctional rearrangement of the hepatic vasculature is shown to play an important role in this process. It is characterized by endothelial dysfunction and impaired paracrine interaction between activated stellate hepatocytes and sinusoidal endotheliocytes, sinusoidal remodeling and capillarization, as well as intrahepatic angiogenesis.


Opioid antagonist (naltrexone) modulation of cerebellar development: histological and morphometric studies.
The role of endogenous opioid systems in preweaning cerebellar development was explored in rats utilizing naltrexone, a potent opioid antagonist. Sprague-Dawley rats were given daily subcutaneous injections of either 1 or 50 mg/kg naltrexone to invoke a temporary or complete blockade, respectively, of opioid receptors throughout the first 3 weeks of postnatal life; animals injected with sterile water served as controls. At weaning (day 21), macroscopic, morphometric, and histological determinations were conducted. In general, 50 mg/kg naltrexone had a stimulatory action on cerebellar development, whereas 1 mg/kg naltrexone had an inhibitory influence. Both sexes were affected comparably. Limits to naltrexone's ability to modulate cerebellar ontogeny were noted, with more latitude existing toward growth enhancement than impairment. The temporal course of ontogeny was generally unaltered in naltrexone-treated rats. Rather, only events that transpired over this period were dramatically affected. The most notable effects in 1 mg/kg naltrexone rats were marked decreases in cerebellar areal dimensions, the content of internal granule neurons, and cellular and tissue differentiation. Characteristics of the 50 mg/kg naltrexone group included increases in cerebellar areal dimensions, neural cell number, content, and size, and structural changes consistent with acceleration in growth and differentiation. Naltrexone influenced both neurons and glia, but only neural cells still being generated during the first 21 d after birth were altered in regard to quantity. Previous evidence has shown the presence of peak levels of endogenous opioids and opioid receptors in the cerebellum during the first weeks of life, as well as demonstrating the presence of enkephalin immunoreactivity on germinative cerebellar cells during postnatal neurogenesis but not in adult counterparts.(ABSTRACT TRUNCATED AT 250 WORDS)


Real-time adaptive contrast enhancement.
A recursive filter approach is introduced to simplify real-time implementation of an adaptive contrast enhancement scheme for imaging sensors. With this scheme, even scenes possessing large global dynamic ranges (>40 dB) can be accommodated by the limited dynamic range (20 dB) of a display without losing the local contrast essential for image interpretation. This paper describes the recursive filter implementation of the local area contrast enhancement scheme using charge-coupled devices and the resultant real-time hardware capable of processing standard 525 and 875 line TV compatible video (from vidicons, videotape recorders, etc). Several examples from video imagery are included to demonstrate its effectiveness.


Genetic analysis workshop 13: Summary of analyses of alcohol and cigarette use phenotypes in the Framingham Heart Study.
Data from the Framingham Heart Study were provided for Genetic Analysis Workshop 13. This paper summarizes six contributed papers that focused on the analysis of the longitudinal alcohol and/or cigarette use phenotypes available in these data, with the goal of detecting genes influencing these traits. For several of these contributions, the primary phenotype was maximum daily substance use reported over the longitudinal data. Others focused on a cross section by taking the daily use reported at a fixed time interval for all individuals, and others considered dichotomous phenotypes defined by thresholds of use. A variety of covariates were considered, including age (or year of birth) and sex. Most studies included unexposed individuals (those reporting no alcohol/cigarette use at all available assessments) in the linkage analyses, though one study compared results when defining the phenotype for unexposed individuals to be zero vs. defining the phenotype for unexposed individuals to be unknown. Linkage findings varied across studies. However, there was some concordance of evidence on chromosome 9 for alcohol traits, and of weaker evidence on chromosome 20 for cigarette traits. Analytical issues arose which may be crucial for genetic studies of substance use and dependence, including the choice of how to handle unexposed or substance-naive individuals, use of dichotomous vs. quantitative traits, and consideration of transformations and covariates.


Cognitive impairment in patients with diabetes.
Type land type 2 diabetes are associated with major complications that shorten life expectancy. Fluctuations in the amount of glucose reaching the brain cause a wide range of alterations to cerebral function, from mild impairment through to dementia-like states and even death. The responsibility is on the patient with diabetes to control his or her blood glucose levels to reduce any mental decline. However, nursing staff should be aware that patients with diabetes experiencing a loss of mental function may have difficulty in understanding information and instructions. This article examines the current literature on loss of mental function as a chronic complication of diabetes.


Canadian Registry of LVIS Jr for Treatment of Intracranial Aneurysms (CaRLA).
Stents reduce the rate of angiographic recurrence of intracranial aneurysms. The newest stent for intracranial use is the Low-profile Visible Intraluminal Support device (LVIS Jr). To assess the efficacy of the new stent in a multicenter retrospective registry. Centers across Canada using LVIS Jr were contacted and asked to participate in a retrospective registry of consecutive patients treated with LVIS Jr for intracranial aneurysms between January 2013 and July 2015. A total of 102 patients, with saccular aneurysms in 100 patients (72 women; age range 21-78 years; mean 56.0 years; median 57.5 years) were treated with a LVIS Jr stent. The mean maximum diameter of the dome and neck of the aneurysm and dome to neck ratios were 8.3 mm±7.7 mm, 4.4 mm±1.9 mm, and 1.86±1.22, respectively. Angiographic complications arose in 23 patients, clinical complications in 9 patients, and only 3% of permanent neurological deficits occurred. Death occurred in 1 patient, unrelated to the stent. The ruptured status of the aneurysms (OR=3.29; p=0.046) and use of LVIS Jr for bailout (OR=2.54; p=0.053) showed a trend towards significant association with higher angiographic complications. At the last available follow-up, 68 class I, 20 class II, and 12 class III results were seen. The LVIS Jr stent is a safe and effective device for stent-assisted coiling, with 3% permanent neurological complications. Stent-assisted coiling continues to be technically challenging in cases of ruptured aneurysms and bailout situations.


Intravenous infusion of human bone marrow mesenchymal stromal cells promotes functional recovery and neuroplasticity after ischemic stroke in mice.
Transplantation of human bone marrow mesenchymal stromal cells (hBM-MSC) promotes functional recovery after stroke in animal models, but the mechanisms underlying these effects remain incompletely understood. We tested the efficacy of Good Manufacturing Practices (GMP) compliant hBM-MSC, injected intravenously 3.5 hours after injury in mice subjected to transient middle cerebral artery occlusion (tMCAo). We addressed whether hBM-MSC are efficacious and if this efficacy is associated with cortical circuit reorganization using neuroanatomical analysis of GABAergic neurons (parvalbumin; PV-positive cells) and perineuronal nets (PNN), a specialized extracellular matrix structure which acts as an inhibitor of neural plasticity. tMCAo mice receiving hBM-MSC, showed early and lasting improvement of sensorimotor and cognitive functions compared to control tMCAo mice. Furthermore, 5 weeks post-tMCAo, hBM-MSC induced a significant rescue of ipsilateral cortical neurons; an increased proportion of PV-positive neurons in the perilesional cortex, suggesting GABAergic interneurons preservation; and a lower percentage of PV-positive cells surrounded by PNN, indicating an enhanced plastic potential of the perilesional cortex. These results show that hBM-MSC improve functional recovery and stimulate neuroprotection after stroke. Moreover, the downregulation of "plasticity brakes" such as PNN suggests that hBM-MSC treatment stimulates plasticity and formation of new connections in the perilesional cortex.


Initial uptake, time to treatment, and real-world effectiveness of all-oral direct-acting antivirals for hepatitis C virus infection in the United States: A retrospective cohort analysis.
Data on initiation and utilization of direct-acting antiviral therapies for hepatitis C virus infection in the United States are limited. This study evaluated treatment initiation, time to treatment, and real-world effectiveness of direct-acting antiviral therapy in individuals with hepatitis C virus infection treated during the first 2 years of availability of all-oral direct-acting antiviral therapies. A retrospective cohort analysis was undertaken using electronic medical records and chart review abstraction of hepatitis C virus-infected individuals aged >18 years diagnosed with chronic hepatitis C virus infection between January 1, 2014, and December 31, 2015 from the Indiana University Health database. Eight hundred thirty people initiated direct-acting antiviral therapy during the 2-year observation window. The estimated incidence of treatment initiation was 8.8%±0.34% at the end of year 1 and 15.0%±0.5% at the end of year 2. Median time to initiating therapy was 300 days. Using a Cox regression analysis, positive predictors of treatment initiation included age (hazard ratio, 1.008), prior hepatitis C virus treatment (1.74), cirrhosis (2.64), and history of liver transplant (1.5). History of drug abuse (0.43), high baseline alanine aminotransferase levels (0.79), hepatitis B virus infection (0.41), and self-pay (0.39) were negatively associated with treatment initiation. In the evaluable population (n = 423), 83.9% (95% confidence interval, 80.1-87.3%) of people achieved sustained virologic response. In the early years of the direct-acting antiviral era, <10% of people diagnosed with chronic hepatitis C virus infection received direct-acting antiviral treatment; median time to treatment initiation was 300 days. Future analyses should evaluate time to treatment initiation among those with less advanced fibrosis.


Post-transcriptional nucleotide addition is responsible for the formation of the 5' terminus of histidine tRNA.
All sequenced histidine tRNAs have one additional nucleotide at the 5' end when compared to other tRNA species. Sequence analysis of histidine tRNA genes from Drosophila melanogaster and Schizosaccharomyces pombe showed that the terminal guanylate residue of the mature tRNAs is not encoded by the genes. Analysis of the products from in vitro transcription of these genes in extracts from Drosophila Kc cells demonstrated that the 5'-terminal nucleotide present in the mature tRNA is added post-transcriptionally. The addition reaction requires ATP. A portion of the mature tRNAs are then modified at the 5'-terminal pG. Analysis of the RNA species formed during the in vitro maturation of the Drosophila histidine tRNA primary transcript uncovered the following maturation scheme: (i) the primary transcript is processed by RNase P at the 5' end to form an intermediate precursor; (ii) the 3'-flanking sequence is endonucleolytically removed, and a guanylate moiety is added to the 5' end to form mature-sized histidine tRNA; and (iii) a fraction of the 5'-terminal guanylate residues then undergoes modification. In contrast to the capping of eukaryotic mRNA, the guanylate addition to histidine tRNA results in the formation of a (3'-5')-phosphodiester bond. There are no precedents for the post-transcriptional addition of nucleotides (in phosphodiester linkage) to the 5' end of RNA precursors.


The role of echocardiographic screening in reducing the burden of rheumatic heart disease in Latin America.
The objectives of this article are to reflect on the rationale behind the use of echocardiographic screening for rheumatic heart disease and to provide key recommendations about steps needed to implement and improve echocardiographic screening programs in Latin America. Rheumatic heart disease remains a public health problem affecting mainly low-income and lower-middle-income countries and populations. Latin America is an area with economic inequalities, and the epidemiology of rheumatic heart disease remains largely unknown. Echocardiographic screening is useful for updating the epidemiology and providing early diagnosis of the disease. We discuss different approaches used in successful echocardiographic screening programs worldwide and in Latin America. We then identify the key elements needed to establish successful echocardiographic screening programs in Latin America, including increased awareness and involvement from multiple sectors (e.g. the community, health care professionals, scientific organizations and public health entities), identification of areas in need, development of a plan and structure that include different screening approaches, and how to ensure appropriate follow up for those who screen positive.


Specific amino acids inhibit food intake via the area postrema or vagal afferents.
To maintain nutrient homeostasis the central nervous system integrates signals that promote or inhibit eating. The supply of vital amino acids is tuned by adjusting food intake according to its dietary protein content. We hypothesized that this effect is based on the sensing of individual amino acids as a signal to control food intake. Here, we show that food intake was most potently reduced by oral L-arginine (Arg), L-lysine (Lys) and L-glutamic acid (Glu) compared to all other 17 proteogenic amino acids in rats. These three amino acids induced neuronal activity in the area postrema and the nucleus of the solitary tract. Surgical lesion of the area postrema abolished the anorectic response to Arg and Glu, whereas vagal afferent lesion prevented the response to Lys. These three amino acids also provoked gastric distension by differentially altering gastric secretion and/or emptying. Importantly, these peripheral mechanical vagal stimuli were dissociated from the amino acids' effect on food intake. Thus, Arg, Lys and Glu had a selective impact on food processing and intake suggesting them as direct sensory input to assess dietary protein content and quality in vivo. Overall, this study reveals novel amino acid-specific mechanisms for the control of food intake and of gastrointestinal function.


Factor VII deficiency detected in pregnancy: a case report.
Factor VII deficiency is a rare hereditary coagulation disorder with an incidence estimated at 1 in 500,000 individuals. In this report, we describe the 13th case in pregnancy. The diagnosis of severe factor VII deficiency (factor VII level <5%) was established at 10 weeks' gestation after initial laboratory testing showed a markedly prolonged prothrombin time and a normal activated partial thromboplastin time. There was a history of two preterm deliveries, but there was no evidence of previous bleeding manifestations. Antenatal progress of the index pregnancy was unremarkable. Prophylactic treatment with fresh frozen plasma was started at the onset of labor and the patient had a vaginal delivery of a live girl at 36 weeks' gestation. There was no postpartum hemorrhage and mother and newborn were discharged in good condition. The patient's postpartum level of factor VII remained undetectable. Two aspects are outlined: the absence of any significant increase in factor VII clotting activity during this pregnancy and the need to give replacement therapy at labor in patients with severe factor VII deficiency to decrease the risk of postpartum hemorrhage.


Impact of Pregnancy on the Pharmacokinetics and Metabolism of Nicotinamide in Humans.
In preeclampsia models, nicotinamide (NAM) has protective effects in preeclampsia and is being evaluated as a therapeutic nutraceutical in clinical studies. NAM undergoes extensive hepatic metabolism by NAM N-methyltransferase to methylnicotinamide (MNA), which is subsequently metabolized to methyl-2-pyridone-5-carboxamide (M2PY) by aldehyde oxidase. However, the pharmacokinetics (PK) of NAM and its major metabolites has never been studied in pregnant individuals. Blood samples were collected before and 1, 2, 4, 8, and 24 hr after single 1g oral NAM dose in healthy pregnant (gestational age 24-33 weeks) and nonpregnant female volunteers (n=6/group). Pooled urine was collected from 0-8 hr. NAM, MNA and M2PY area under the concentration-time curve (AUC) data were analyzed by noncompartmental analysis. No difference in the plasma AUC0→24 of NAM (median [25%-75%]: 463 [436-576] vs 510 [423, 725] μM*hr, p=0.430) and its intermediate metabolite MNA (89.1 [60.4, 124.4] vs 83.8 [62.7, 93.7] μM*hr, p=0.515) was observed in pregnant and nonpregnant volunteers, respectively; however, the terminal metabolite M2PY AUC0→24 was significantly lower in pregnant individuals (218 [188, 254] vs 597 [460, 653] μM*hr, p<0.001). NAM renal clearance (CLR ) (p=0.184), MNA CLR (p=0.180), and total metabolite formation clearance (CLform ) (p=0.405) did not differ across groups; however, M2PY CLR was significantly higher in pregnant individuals (10.5 [9.3-11.3] vs 7.5 [6.4-8.5] L/hr, p=0.002). These findings demonstrate that the PK of NAM and systemic exposure to its intermediate metabolite MNA are not significantly altered during pregnancy, and systemic exposure to NAM's major metabolite M2PY was reduced during pregnancy due to increased renal elimination.


Quality assurance of MLC leaf position accuracy and relative dose effect at the MLC abutment region using an electronic portal imaging device.
We investigated an electronic portal image device (EPID)-based method to see whether it provides effective and accurate relative dose measurement at abutment leaves in terms of positional errors of the multi-leaf collimator (MLC) leaf position. A Siemens ONCOR machine was used. For the garden fence test, a rectangular field (0.2 20 cm) was sequentially irradiated 11 times at 2-cm intervals. Deviations from planned leaf positions were calculated. For the nongap test, relative doses at the MLC abutment region were evaluated by sequential irradiation of a rectangular field (2 20 cm) 10 times with a MLC separation of 2 cm without a leaf gap. The integral signal in a region of interest was set to position A (between leaves) and B (neighbor of A). A pixel value at position B was used as background and the pixel ratio (A/B 100) was calculated. Both tests were performed at four gantry angles (0, 90, 180 and 270°) four times over 1 month. For the nongap test the difference in pixel ratio between the first and last period was calculated. Regarding results, average deviations from planned positions with the garden fence test were within 0.5 mm at all gantry angles, and at gantry angles of 90 and 270° tended to decrease gradually over the month. For the nongap test, pixel ratio tended to increase gradually in all leaves, leading to a decrease in relative doses at abutment regions. This phenomenon was affected by both gravity arising from the gantry angle, and the hardware-associated contraction of field size with this type of machine.


Efficacy of anti-T-lymphocyte globulin-Fresenius as an induction agent in deceased-donor renal transplantation: A cohort study.
Anti-T-lymphocyte globulin (ATG) is frequently used in the induction regimen of renal transplantation, but its dose has not been standardized. In the present study, the efficacy of different ATG-Fresenius (ATG-F) doses was assessed in recipients of renal transplantation. A total of 131 adult recipients of renal transplantation who received ATG-F induction between August 2015 and July 2018 were included. The incidence of biopsy-confirmed acute rejection, graft function, as well as graft and patient survival within 12 months post-transplant, was assessed, and adverse events, including hematologic and infection-associated side effects, were compared between patients receiving a cumulative ATG-F dose of <7 or ≥7 mg/kg. The incidence of biopsy-confirmed acute rejection was similar between patients receiving cumulative doses of <7 and ≥7 mg/kg (7.5 vs. 4.7%, P=0.766). The incidence of infection within 12 months was lower in the ATG-F <7 mg/kg group compared with that in the ≥7 mg/kg group (26.9 vs. 50.0%, P=0.006), but the incidence of pneumonia did not differ between the ATG-F <7 and ≥7 mg/kg groups (10.4 vs. 20.3%, P=0.117). The incidence of urinary infection was higher in the ≥7 mg/kg group than in the <7 mg/kg group (20.4 vs. 7.46%, P=0.033), while the extent and duration of anemia and lymphopenia was similar between groups. There was no difference in graft function, delayed graft function, as well as overall and graft survival between the groups. In conclusion, a moderate reduction in the cumulative ATG-F dose was not associated with an increased risk of acute rejection, while the risk of infection was reduced. Optimization of the ATG-F dose for induction may facilitate the reduction of the risk of infection without compromising the induction efficacy in renal transplant recipients.


The influence of chromatin structure on DNA damage induced by nitrogen mustard and cisplatin analogues.
The interaction of anti-tumour drugs with reconstituted chromatin has been investigated using defined nucleosomal complexes. This allowed the effect of nucleosome cores on drug-induced DNA damage to be assessed for four nitrogen mustard analogues, dimethylsulphate and three cisplatin analogues. A defined nucleosomal complex was employed that contained two precisely positioned nucleosome cores. The construct was then subjected to drug treatment, and the resulting DNA damage was quantitatively analysed using a Taq DNA polymerase stop assay. At the sites of damage, densitometric comparisons between purified and reconstituted DNA were used to evaluate the influence of nucleosomal core proteins on specific drug-DNA interactions. Results were combined with previous data obtained for other DNA-damaging drugs investigated using the same nucleosomal construct. For most of the DNA-damaging agents studied, this method revealed protection at the positioned nucleosome cores and indicated that the preferred site of DNA binding for these compounds was in the linker region of the construct. Statistical analyses confirmed the significant level of damage protection conferred by the nucleosome cores and revealed differences between the examined compounds. Larger compounds generally displayed a greater tendency to target the linker region of the nucleosomal DNA and were impeded from damaging nucleosomal core DNA. In contrast, smaller molecules had greater access to nucleosomal core DNA.


Differential functional avidity of dengue virus-specific T-cell clones for variant peptides representing heterologous and previously encountered serotypes.
Proinflammatory cytokines secreted by memory CD8+ and CD4+ T cells are thought to play a direct role in the pathogenesis of dengue virus infection by increasing vascular permeability and thereby inducing the pathophysiologic events associated with dengue hemorrhagic fever and dengue shock syndrome. Severe disease is frequently observed in the setting of secondary infection with heterologous dengue virus serotypes, suggesting a role for cross-reactive memory T cells in the immunopathogenesis of severe disease. We used a large panel of well-characterized dengue virus-specific CD8+ T-cell clones isolated from Pacific Islanders previously infected with dengue virus 1 to examine effector memory function, focusing on a novel dominant HLA-B*5502-restricted NS5(329-337) epitope, and assessed T-cell responses to stimulation with variant peptides representing heterologous serotypes. Variant peptides were differentially recognized by dengue virus 1-specific effector CD8+ cytotoxic T lymphocytes (CTL) in a heterogeneous and clone-specific manner, in which cytolytic function and cytokine secretion could be enhanced, diminished, or abrogated compared with cognate peptide stimulation. Dengue virus-specific CTL stimulated with cognate and variant peptides demonstrated a cytokine response hierarchy of gamma IFN (IFN-gamma) > tumor necrosis factor alpha (TNF-alpha) > interleukin-2 (IL-2), and a subset of clones also produced IL-4 and IL-6. Individual clones demonstrated greater avidity for variant peptides representing heterologous serotypes, including serotypes previously encountered by the subject, and IFN-gamma and TNF-alpha secretion was enhanced by stimulation with these heterologous peptides. Altered antiviral T-cell responses in response to stimulation with heterologous dengue virus serotypes have implications for control of virus replication and for disease pathogenesis.


[The role of weak specific and nonspecific interactions in recognition and conversation by enzymes of long DNA].
According to a currently accepted model, enzymes engage in high-rate sliding along DNA when searching for specific recognition sequences or structural elements (modified nucleotides, breaks, single-stranded DNA fragments, etc.). Such sliding requires these enzymes to possess sufficiently high affinity for DNA of any sequence. Thus, significant differences in the enzymes' affinity for specific and nonspecific DNA sequences cannot be expected, and formation of a complex between an enzyme and its target DNA unlikely contributes significantly in the enzyme specificity. To elucidate the factors providing the specificity we have analyzed many DNA replication, DNA repair, topoisomerization, integration, and recombination enzymes using a number of physicochemical methods, including a method of stepwise increase in ligand complexity developed in our laboratory. It was shown that high affinity of all studied enzymes for long DNA is provided by formation of many weak contacts of the enzymes with all nucleotide units covered by protein globules. Contacts of positively charged amino acid residues with internucleotide phosphate groups contribute most to such interactions; the contribution of each contact is very small and the full contact interface usually resembles interactions between oppositely charged biopolymer surfaces. In some cases significant contribution to the affinity is made through hydrophobic and/or van der Waals interactions of the enzymes with nucleobases. Overall, depending on the enzyme, such nonspecific interactions provide 5-8 orders of the enzyme affinity for DNA. Specific interactions of enzymes with long DNA, in contrast to contacts of enzymes with small ligands, are usually weak and comparable in efficiency with weak nonspecific contacts. The sum of specific interactions most often provides approximately one and rarely two orders of the affinity. According to structural data, DNA binding to any of the investigated enzymes is followed by a stage of DNA conformation adjustment including partial or complete DNA melting, deformation of its backbone, stretching, compression, bending or kinking, eversion of nucleotides from the DNA helix, etc. The full set of such changes is characteristic for each individual enzyme. The fact that all enzyme-dependent changes in DNA are effected through weak specific rather than strong interactions is very important. Enzyme-specific changes in DNA conformation are required for effective adjustment of reacting orbitals with accuracy about 10-15 degrees, which is possible only for specific DNA. A transition from nonspecific to specific DNA leads to an increase in the reaction rate (kcat) by 4-8 orders of magnitude. Thus, the stages of DNA conformation adjustment and catalysis proper provide the high specificity of enzyme action.


Caffeine choice prospectively predicts positive subjective effects of caffeine and d-amphetamine.
Individuals vary in their subjective and behavioral response to psychomotor stimulants and these differences may be associated with the likelihood of developing problematic use of these drugs. The present study sought to determine whether individual differences in caffeine choice prospectively predict subjective response to acute doses of caffeine and d-amphetamine. In Phase 1, Choosers and Nonchoosers of caffeine were identified using 10 independent choice trials in which subjects repeatedly chose between caffeine (200mg/70 kg) and placebo. Choosers were defined as those who chose caffeine over placebo on ≥7 of the 10 trials; Nonchoosers were those who chose placebo on ≥7 trials. In Phase 2, Choosers and Nonchoosers were compared in their subjective response to caffeine (100, 200, 400mg/70 kg) and d-amphetamine (5, 10, 20 mg/70 kg). Of the 22 participants completing the study, 11 met criteria for being a caffeine Chooser and 8 were Nonchoosers. In Phase 1, Choosers reported higher ratings of positive (i.e., pleasant) and lower ratings of negative (i.e., unpleasant) effects of caffeine during the sampling sessions. In Phase 2, caffeine Choosers reported more positive subjective effects and fewer negative effects of caffeine and d-amphetamine, particularly at the highest doses examined. Individual differences in caffeine reinforcement predicted subsequent subjective response to both d-amphetamine and caffeine. This observation may have clinical utility for identifying individuals who are vulnerable to the reinforcing effects of abused psychomotor stimulants.


Compassion Fatigue Among RNs Working in Skilled Nursing Facilities.
Nurses employed in skilled nursing facilities (SNFs) are at risk for developing compassion fatigue. Compassion fatigue negatively impacts patient outcomes and is associated with decreased quality of care. Some nurses leave the profession due to compassion fatigue. There are no published studies in the United States focused on compassion fatigue among SNF nurses. The purpose of this interpretive phenomenological study was to identify the shared meanings of compassion fatigue among RNs caring for residents in SNFs. Shared meanings among participants were identified as: (1) I feel conflicted, which causes my compassion fatigue; (2) I feel physical and emotional manifestations of compassion fatigue; (3) Compassion fatigue is infused in every aspect of my life; and (4) We are trying to cope with compassion fatigue. Of paramount importance was the desire to make a difference in the lives of residents. Three participants sought employment outside the SNF setting due to compassion fatigue. Further research and education are needed to improve knowledge, policy, and practice. [Research in Gerontological Nursing, 13(6), 320-328.].


What determines sub-diffusive behavior in crowded protein solutions?
The aqueous environment inside cells is densely packed. A typical cell has a macromolecular concentration in the range 90-450 g/L, with 5-40% of its volume being occupied by macromolecules, resulting in what is known as macromolecular crowding. The space available for the free diffusion of metabolites and other macromolecules is thus greatly reduced, leading to so-called excluded volume effects. The slow diffusion of macromolecules under crowded conditions has been explained using transient complex formation. However, sub-diffusion noted in earlier works is not well characterized, in particular, the role played by transient complex formation and excluded volume effects. We have used Brownian dynamics simulations to characterize the diffusion of chymotrypsin inhibitor 2 (CI2) in protein solutions of bovine serum albumin (BSA) and lysozyme at concentrations ranging from 50 to 300 g/L. The predicted changes in diffusion coefficient as a function of crowder concentration are consistent with NMR experiments. The sub-diffusive behavior observed in the sub-microsecond time scale can be explained in terms of a so-called cage effect, arising from rattling motion in a local molecular cage as a consequence of excluded volume effects. By selectively manipulating the nature of interactions between protein molecules, we determined that excluded volume effects induce sub-diffusive dynamics at sub-microsecond time scales. These findings may help to explain the diffusion-mediated effects of protein crowding on cellular processes.


Prism adaptation treatment to address spatial neglect in an intensive rehabilitation program: A randomized pilot and feasibility trial.
Spatial neglect (SN) is a common cognitive disorder after brain injury. Prism adaptation treatment (PAT) is one of the promising interventions for SN albeit inconsistent results from previous studies. We carried out a comparison intervention (PAT vs. Sham) and aimed to evaluate the efficacy of PAT on visuospatial symptoms of SN in an inpatient rehabilitation setting that offered a highly intensive comprehensive brain injury rehabilitation program. A total of 34 patients with moderate-to-severe SN secondary to stroke or traumatic brain injury were randomized to the PAT group and the Sham group (an active control group). Both groups received 10 sessions of treatment, over two weeks, in addition to the rehabilitation therapies provided by their rehabilitation care teams. Outcomes were measured using an ecological instrument (the Catherine Bergego Scale) and paper-and-pencil tests (the Bells Test, the Line Bisection Test and the Scene Copying Test). Patients were assessed at baseline, immediately after treatment, two weeks after treatment, and four weeks after treatment. 23 (67.6%) patients completed treatment and all the assessment sessions and were included in the final analyses using mixed linear modeling. While SN symptoms reduced in both groups, we found no difference between the two groups in the degree of improvement. In addition, the average SN recovery rates were 39.1% and 28.6% in the PAT and Sham groups, respectively, but this discrepancy did not reach statistical significance. Thus, the present study suggests that PAT may contribute little to SN care in the context of a highly intensive inpatient rehabilitation program. Further large-scale investigation is required to uncover the mechanisms underlying PAT and Sham in order to refine the treatment or create new interventions.


Anti hyperglycemic activities of Annona muricata (Linn).
This study was designed to determine the effects of methanolic extracts of Annona muricata (Linn) on the blood glucose level of streptozotocin-induced diabetic Wistar rats. Thirty adult Wistar rats were randomly assigned into three groups (A, B and C) of ten rats each. Group A was the control, Group B was untreated hyperglycemic group and group C was A. muricata-treated group. Hyperglycemia was induced in groups B and C by a single intraperitoneal injection of 80 mg/kg streptozotocin dissolved in 0.1 M citrate buffer. The control group was intraperitoneally injected with equivalent volume of citrate buffer and all the animals were monitored for four weeks. Daily intra peritoneal injection of 100 mg/kg A. muricata was administered to group C rats for two weeks and the animals were monitored for another two weeks. The data obtained were analyzed with descriptive and inferential statistics. The result showed a mean body weight of 206 +/- 7.74 g, 173.29+/-5.13 g and 197 +/- 5.62 g respectively for the control, untreated diabetic and A. muricata-treated diabetic group, and a mean blood glucose concentration of 3.78 +/- 0.190 mmol/L, 21.64 +/- 2.229 mmol/L and 4.22 +/- 0.151 mmol/L for the control, untreated diabetic and treated diabetic groups respectively. A significant difference exists between the blood glucose concentrations of treated and untreated hyperglycemic groups of rats. The result of this study demonstrated that A. muricata possesses anti-hyperglycemic activities.


Wake and Sleep EEG in Patients With Huntington Disease: An eLORETA Study and Review of the Literature.
The aim of the study was to evaluate the EEG modifications in patients with Huntington disease (HD) compared with controls, by means of the exact LOw REsolution Tomography (eLORETA) software. We evaluated EEG changes during wake, non-rapid eye movement (NREM) and rapid eye movement (REM) sleep. Moreover, we reviewed the literature concerning EEG modifications in HD. Twenty-three consecutive adult patients affected by HD were enrolled, 14 women and 9 men, mean age was 57.0 ± 12.4 years. Control subjects were healthy volunteers (mean age 58.2 ± 14.6 years). EEG and polygraphic recordings were performed during wake (before sleep) and during sleep. Sources of EEG activities were determined using the eLORETA software. In wake EEG, significant differences between patients and controls were detected in the delta frequency band (threshold T = ±4.606; P < .01) in the Brodmann areas (BAs) 3, 4, and 6 bilaterally. In NREM sleep, HD patients showed increased alpha power (T = ±4.516; P < .01) in BAs 4 and 6 bilaterally; decreased theta power (T = ±4.516; P < .01) in the BAs 23, 29, and 30; and decreased beta power (T = ±4.516; P < .01) in the left BA 30. During REM, HD patients presented decreased theta and alpha power (threshold T = ±4.640; P < .01) in the BAs 23, 29, 30, and 31 bilaterally. In conclusion, EEG data suggest a motor cortex dysfunction during wake and sleep in HD patients, which correlates with the clinical and polysomnographic evidence of increased motor activity during wake and NREM, and nearly absent motor abnormalities in REM.


[Policy framework for the protection of integrity in research in Brazil].
Beginning with the assumption that the heated tension between research freedom and the protection of human life and integrity has not been overcome, this article discusses the issue of regulatory framework for the protection of research integrity in the Brazilian legal system. Throughout the work, the general aspects and principles that establish the limits between scientific activity and the rights and interests of the subjects of experimental processes are developed, as well as the treatment deserved by certain especially problematic situations derived from the complex cultural composition of Brazilian society.


Home-environment controls for the mobility-impaired.
One aspect of self-care and independence for many older individuals concerns their ability to control their home environment when their mobility is impaired. Although rehabilitation devices have often been custom designed, the availability of these devices has been limited because of high manufacturing costs and a lack of production and marketing channels. It should therefore be recognized that many devices already available to consumers offer a wide range of environmental control without the problems associated with custom design.


Physiological Effects of Oxidative Stress Caused by Saxitoxin in the Nematode Caenorhabditis elegans.
Saxitoxin (STX) causes high toxicity by blocking voltage-gated sodium channels, and it poses a major threat to marine ecosystems and human health worldwide. Our work evaluated the neurotoxicity and chronic toxicology of STX to Caenorhabditis elegans by an analysis of lifespan, brood size, growth ability, reactive oxygen species (ROS) and adenosine triphosphate (ATP) levels, and the overexpression of green fluorescent protein (GFP). After exposure to a series of concentrations of STX for 24 h, worms showed paralysis symptoms and fully recovered within 6 h; less than 5% of worms died at the highest concentration of 1000 ng/mL for first larval stage (L1) worms and 10,000 ng/mL for fourth larval stage (L4) worms. Declines in lifespan, productivity, and body size of C. elegans were observed under the stress of 1, 10, and 100 ng/mL STX, and the lifespan was shorter than that in controls. With STX exposure, the productivity declined by 32-49%; the body size, including body length and body area, declined by 13-18% and 25-27%, respectively. The levels of ROS exhibited a gradual increase over time, accompanied by a positive concentration effect of STX resulting in 1.14-1.86 times higher levels compared to the control group in L4 worms. Conversely, no statistically significant differences were observed between L1 worms. Finally, after exposure to STX for 48 h, ATP levels and GFP expression in C. elegans showed a significant dose-dependent increase. Our study reports the first evidence that STX is not lethal but imposes substantial oxidative stress on C. elegans, with a dose-responsive relationship. Our results indicated that C. elegans is an ideal model to further study the mechanisms underlying the fitness of organisms under the stress caused by paralytic shellfish toxins including STX.


Colorectal cancer patients' experiences with supervised exercise during adjuvant chemotherapy-A qualitative study.
Colorectal cancer (CRC) is a common cancer worldwide, with increasing numbers surviving and living with long-term side effects from treatment. Physical exercise during or after treatment may have several beneficial effects, but knowledge of CRC patients' reflections on exercising during adjuvant therapy is limited. The aim of this study was to explore the experiences of CRC patients participating in a supervised exercise program during adjuvant chemotherapy. This study included CRC patients participating in two intervention studies with individually tailored and supervised combinations of endurance, resistance, and balance exercises during adjuvant chemotherapy. Semi-structured interviews performed at the beginning, during, and immediately after the intervention period from 15 participants were analyzed using thematic analysis. Four main themes identified were "structuring life with cancer," "motivation to exercise," "training experiences," and "effects of exercise." Scheduled appointments gave structure to daily life and served as an external motivational factor. The individual adjustments of exercise gave a sense of security and helped improving adherence, especially when feeling depressed or fatigued. Common expectations were improvement of endurance and strength and counteracting negative effects of chemotherapy. Experienced positive effects from exercising, both mentally and physically, contributed to inner motivation and inspired continued exercising after the study period. This study offers important insights into CRC patients' experiences of participating in a physical exercise program during adjuvant chemotherapy. Based on our findings, we recommend supervised and individually tailored physical exercise during adjuvant chemotherapy to this patient group.


Assessing the sustainability of contrasting land use scenarios through the Socioecological Integrated Analysis (SIA) of the metropolitan green infrastructure in Barcelona.
Urban development and the sprawl of transport infrastructures have disregarded the crucial function of metropolitan landscape in provisioning human well-being and biodiversity. This research aims to contribute to the challenges of Planning for Sustainability by proposing a Socioecological Integrated Analysis (SIA) to support the Land Use Master Plan in the Barcelona Metropolitan Area, to conciliate urban development with the performance of surrounding open spaces. The paper evaluates four different land cover scenarios (current, trending, alternative and potential), and two kinds of agricultural management (conventional and a socioecological transition towards organic agriculture). The results suggest that although there are significant improvements on job provisioning and nutrient-cycling closures (circular economy), certified organic agriculture is not enough to overcome some trends of industrialized agrarian systems such as low energy efficiency or poor improvements in greenhouse gas emissions. The results also show a crossed effect between social metabolism and landscape ecology where changes in the management could affect the landscape functioning while changes in the land covers are particularly affecting the resource use. Then, deeper changes that consider together land use and metabolic flows are required to promote more sustainable agroecological transitions. The SIA model is an important conceptual and methodological step forward that facilitates the transition towards sustainable land use policies.


Palindromic unit highly repetitive DNA sequences exhibit species specificity within Enterobacteriaceae.
Palindromic units (PU, or REP for repetitive extragenic palindrome) constitute a family of DNA sequences of 40 nucleotides which is highly repeated in the genome of Escherichia coli. We analysed the presence of PU sequences in 99 different bacterial genomes by cross-hybridization. When PU sequences were used as a probe, only DNA from Enterobacteriaceae closely related to E. coli exhibited an appreciable hybridization signal: Shigella sonnei, Shigella boydii, Salmonella enteritica serotype Typhimurium, Citrobacter freundii and Levinea malonatica. Furthermore, these bacteria could be divided into two groups which corresponded to a slight difference in their PU sequence: the E. coli group includes S. sonnei and S. boydii; the S. enteritica serotype Typhimurium group includes C. freundii and L. malonatica.


Response to hepatitis B vaccine in HBsAg/anti-HBs negative and anti-HBc positive subjects.
Some anti-HBc positive subjects have been encountered in the absence of hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody (anti-HBs). The aim of this study was to evaluate the response to hepatitis B vaccination in such cases. A total of 33 subjects who were HBsAg and anti-HBs negative, anti-HBc positive, with normal serum aminotransferase levels were included in the study. A recombinant hepatitis B vaccine was administered to subjects. Sera samples were obtained 1 month after each vaccination and tested for anti-HBs. HBV DNA and HBeAg were not detected in any subject. Anti-HBs levels were measured above 10 > or = mIU/ml in 48.4% of cases after the first vaccination, 63.6% after the second vaccination and 90.9% after the third vaccination. Only 3 subjects (9.1%) lacked antibody response in spite of the 3-dose vaccination. In conclusion, preventive antibody levels were obtained after HBV vaccination in most of the HBsAg, anti-HBs negative, anti-HBc positive persons.


Kava constituents exert selective anticancer effects in oral squamous cell carcinoma cells in vitro.
Kava is a beverage made from the ground roots of the plant Piper Methysticum. Active compounds of Kava have previously been demonstrated to exert an antiproliferative effect through cell cycle arrest and promotion of apoptosis. Our aim was to investigate the in vitro effects of the main constituents derived from Kava on oral squamous cell carcinoma (OSCC) activity. Gas chromatography mass spectrometry (GCMS) was used to characterise the main constituents of two Kava preparations. Cell proliferation was assessed in two human OSCC cell lines (H400 and BICR56) and in normal oral keratinocytes (OKF6) treated with the identified Kava constituents, namely Flavokawain A (FKA), Flavokawain B (FKB), yangonin, kavain and methysticin using an MTS in vitro assay. Cell migration at 16 h was assessed using a Transwell migration assay. Cell invasion was measured at 22 h using a Matrigel assay. Cell adhesion was assessed at 90 min with a Cytoselect Adhesion assay. The two Kava preparations contained substantially different concentrations of the main chemical constituents. Treatment of malignant and normal oral keratinocyte cell lines with three of the identified constituents, 10 μg/ml FKA, 2.5 μg/ml FKB and 10 μg/ml yangonin, showed a significant reduction in cell proliferation in both H400 and BICR56 cancer cell lines but not in normal OKF6 cells. Remarkably, the same Kava constituents induced a significant reduction of OSCC cell migration and invasion. We have demonstrated, for the first time, that Kava constituents, FKA, FKB and yangonin have potential anticancer effects on OSCC. This highlights an avenue for further research of Kava constituents in the development of future cancer therapies to prevent and treat OSCC.


Protective Effects of Melatonin against Severe Burn-Induced Distant Organ Injury: A Systematic Review and Meta-Analysis of Experimental Studies.
Extensive burns result in a local wound response and distant-organ injury (DOI) caused by oxidative-stress and inflammation. Melatonin (MT) shows promise in alleviating oxidative-stress and inflammation, but its role in thermal injury is largely unexplored. The present systematic review and meta-analysis were designed to assess the effects of MT on oxidative-stress and inflammatory markers against severe burn-induced DOI. Mean difference (MD)/standard mean difference (SMD) with 95% confidence interval (CI) were estimated using fixed-effect/random-effects models. Eighteen experimental studies met the inclusion criteria. Compared with the control group, MT significantly decreased the levels of malondialdehyde (SMD, -1.03; 95% CI, -1.30, -0.76, p < 0.00001) and 4-hydroxynonenal (MD, -1.06; 95% CI, -1.57, -0.56, p < 0.0001). Additionally, MT increased the levels of glutathione (SMD, 1.94; 95% CI, 1.27, 2.61, p < 0.00001) and superoxide-dismutase (SMD, 0.76; 95% CI, 0.08, 1.45, p = 0.03). Finally, MT significantly decreased the levels of tumor necrosis factor-α (SMD, -1.34; 95% CI, -1.92 to -0.77; p < 0.00001) and C-reactive protein (MD, -12.67; 95% CI, -16.72 to -8.62; p < 0.00001). Meta-analysis indicates that severe burn followed by immediate MT (10 mg/kg) intervention shows significant beneficial effects after 24-h against DOI by regulating oxidative-stress and the inflammatory response.


Effect of myocardial ischemia and type I antiarrhythmic drug therapy on exercise QT intervals.
Direct measurement of the QT interval during exercise is the preferred method to assess duration of this interval with changes in heart rate. The influence of exercise induced ischemia or type I antiarrhythmic drugs on the exercise QT interval has not been well studied. Exercise electrocardiograms and thallium scintigraphy were performed in three groups-normals, patients with coronary artery disease, and patients taking type I antiarrhythmic drugs. The change in QT interval with heart rate increases during exercise was compared among the three groups. The QT interval shortened progressively in the normal group. Demonstrable myocardial ischemia (positive thallium scan) had no effect on the QT interval. Type I antiarrhythmic drugs prolong the QT interval at rest compared to normals, but the interval shortened to the same degree as in normals during exercise. Thus, normal values can be used to compare QT intervals in patients with coronary artery disease. These values do not apply to patients receiving type I antiarrhythmic drugs.


Successful Treatment of Non-small-cell Lung Cancer with Atezolizumab Following Tubulointerstitial Nephritis Due to Pembrolizumab.
We herein report a 75-year-old man with non-small-cell lung cancer who developed tubulointerstitial nephritis due to pembrolizumab administration. He was successfully treated with atezolizumab following steroid administration. He was initially diagnosed with lung adenocarcinoma (T1bN3M1b, stage IV), with a programmed cell death-ligand 1 tumor proportion score of 25-49%. Although the tumor responded well to pembrolizumab, the drug was discontinued because of the diagnosis of tubulointerstitial nephritis on a renal biopsy. Tubulointerstitial nephritis was treated with 30 mg prednisolone, the dose of which was tapered to and maintained at 5 mg. Following lung cancer progression, atezolizumab was administered, and the tumor responded again. Its efficacy has been sustained for >15 months without recurrence of tubulointerstitial nephritis.


Reactive adsorption of ammonia and ammonia/water on CuBTC metal-organic framework: a ReaxFF molecular dynamics simulation.
We report ReaxFF molecular dynamics simulations for reactive adsorption of NH(3) on dehydrated CuBTC metal-organic framework. If the temperature is moderate (up to 125 °C), the dehydrated CuBTC demonstrates a good hydrostatic stability for water concentrations up to 4.0 molecules per copper site. However, if the temperature increases to 550 K, the dehydrated CuBTC will collapse even at a small water concentration, 1.0 H(2)O molecule per copper site. When NH(3) molecules are adsorbed in the channel and micropores of CuBTC, they prefer to chemisorb to the copper sites rather than forming a dimer with another NH(3) molecule. The formation of equimolar Cu(2)(NH(2))(4) and (NH(4))(3)BTC structures is observed at 348 K, which is in good agreement with previous experimental findings. The dehydrated CuBTC framework is partially collapsed upon NH(3) adsorption, while the Cu-Cu dimer structure remains stable under the investigated conditions. Further calculations reveal that the stability of CuBTC is related to the ammonia concentration. The critical NH(3) concentration after which the dehydrated CuBTC starts to collapse is determined to be 1.0 NH(3) molecule per copper site. Depending on whether NH(3) concentration is below or above the critical value, the dehydrated CuBTC can be stable to a higher temperature, 378 K, or can collapse at a lower temperature, 250 K. H(2)O∕NH(3) mixtures have also been studied, and we find that although water molecules do not demonstrate a strong interaction with the copper sites of CuBTC, the existence of water molecules can substantially prevent ammonia from interacting with CuBTC, and thus reduce the amount of chemisorbed NH(3) molecules on CuBTC and stabilize the CuBTC framework to some extent.


Unexpected death on the non-ICU trauma ward.
To characterize trauma patients who die unexpectedly on the ward (unexpected ward deaths = UWDs), 1,011 trauma-related deaths occurring at a level I trauma center over a 10-year period were reviewed for location of death. Seventy-four deaths occurred on the non-ICU trauma ward (i.e., nonmonitored med-surg floor). Fifty patients were "do not resuscitate" (expected deaths). Twenty-four patients (mean age, 58.0 years) died unexpectedly (2.4% of trauma-related deaths). The majority had a central nervous system injury or a precipitating event that occurred at night. Twelve (50%) of the UWDs were determined by peer review to be potentially preventable and were the result of delayed diagnosis (n = 6), aspiration (n = 3), or cardiorespiratory arrest (n = 3). We conclude that unexpected trauma center deaths related to events occurring on the non-ICU trauma ward (2.4% of trauma deaths) occur mostly at night in older, neurologically impaired patients and that half of these deaths may be potentially preventable. Increased awareness of this issue and an environment for direct patient observation may reduce the number of these potentially preventable deaths.


Biodegradability of industrial textile wastewater - batch tests.
Following new trends we applied oxygen uptake rate (OUR) tests as well as long-term tests (in two batch bioreactors systems) in order to assess the biodegradability of textile wastewater. Effluents coming from a dyeing factory were divided into two streams which differed in inorganic and organic contaminants loads. Usefulness of the stream division was proved. Biodegradation of the low-loaded stream led to over 97% reduction of biochemical oxygen demand (BOD5) together with 80% reduction of chemical oxygen demand (COD) and total organic carbon (TOC). Most of the controlled parameter values were below the levels allowed by legislation for influents to surface water, whereas the high-loaded stream was so contaminated with recalcitrant organic compounds that despite the reduction of BOD5 by over 95%, COD, TOC, total nitrogen and total phosphorus levels exceeded permissible values. OUR tests were aimed at determination of the following kinetic parameters: maximum specific growth rate (μMax), half-saturation constant, hydrolysis constant and decay coefficient for activated sludge biomass for both types of textile wastewater studied. The values of kinetic parameters will be applied in activated sludge models used for prediction and optimisation of biological treatment of textile wastewater.


LINC00869 promotes hepatocellular carcinoma metastasis via protrusion formation.
Coordination of filament assembly and membrane remodeling is required for the directional migration of cancer cells. The Wiskott-Aldrich syndrome protein (WASp) recruits the actin-related protein (Arp) 2/3 complex to assemble branched actin networks. The goal of our study was to assess the potential regulatory role exerted by the novel long non-coding RNA (lncRNA) LINC00869 on hepatocellular carcinoma (HCC) cells. We used HCC cells to overexpress or knockdown LINC00869, analyzed patient data from publicly available databases and Cancer Hospital Affiliated with Zhengzhou University, and used a xenograft mouse model of HCC to study the molecular mechanism associated with LINC00869 expression. We found that high levels of LINC00869 expression were associated with poor prognosis in HCC patients. Next, we detected an interaction between LINC00869 and both WASp and Arp2 in HCC cells, and observed a modulatory effect of LINC00869 on the phosphorylation of WASp at Y291 and the activity of cell division control protein 42 (Cdc42). These modulatory roles were required for WASp/Cdc42 activity on F-actin polymerization to enhance membrane protrusion formation and maintain persistent cell polarization. This, in turn, promoted the migration and invasion abilities of HCC cells. Finally, we confirmed the role of LINC00869 in vivo, using the tumor xenograft mouse model; and identified a positive correlation between LINC00869 expression levels and the phosphorylation levels of WASp in HCC samples. Overall, our findings suggest a unique mechanism by which LINC00869 orchestrates membrane protrusion during migration and invasion of HCC cells. Implications: LncRNA LINC00869 regulates the activity of Cdc42-WASp pathway and positively affects protrusion formation in HCC cells, which expands the current understanding of lncRNA functions as well as gives a better understanding of carcinogenesis.


Rapid detection of pathogenic bacteria and screening of phage-derived peptides using microcantilevers.
We report the use of an array of microcantilevers to measure the specific binding of Salmonella to peptides derived from phage display libraries. Selectivity of these phage-derived peptides for Salmonella spp. and other pathogens ( Listeria monocytogenes and Escherichia coli ) are compared with a commercially available anti- Salmonella antibody and the antimicrobial peptide alamethicin. A Langmuir isotherm model was applied to determine the binding affinity constants of the peptides to the pathogens. One particular peptide, MSal 020417, demonstrated a higher binding affinity to Salmonella spp. than the commercially available antibody and is able to distinguish among eight Salmonella serovars on a microcantilever. A multiplexed screening system to quickly determine the binding affinities of various peptides to a particular pathogen highly improves the efficiency of the peptide screening process. Combined with phage-derived peptides, this microcantilever-based technique provides a novel biosensor to rapidly and accurately detect pathogens and holds potential to be further developed as a screening method to identify pathogen-specific recognition elements.


Bacteriolysis and inhibition of gram-positive bacteria by components of Streptococcus zymogenes lysin.
The ability of the A and L components of Streptococcus zymogenes lysin to cause lysis or to inhibit growth of a variety of gram-positive bacteria has been examined. Frank lysis of some, but not all, strains of S. faecalis, S. faecium, and S. liquefaciens by L component alone was demonstrated. None of the strains of these species was lysed by A component alone. S. durans was not lysed by either component. Inhibition of growth of all enterococcal strains by both A substance-producing and L substance-producing mutants of S. zymogenes was also demonstrated. However, inhibition by the A substance producer was markedly less than by the L substance producer. Inhibition of the growth of a number of other gram-positive genera by both A and L mutants was also noted.