Plasmid-mediated quinolone resistance conferred by qnrS1 in Salmonella enterica serovar Virchow isolated from Turkish food of avian origin.
To study the molecular characteristics of the quinolone and associated ampicillin resistance mechanisms present in Salmonella enterica serovar Virchow isolated from Turkish foods. Nine epidemiologically unrelated Salmonella Virchow strains isolated from foods (chicken and minced meat) sold in different markets in Ankara were analysed for their susceptibility to 17 antimicrobials. The strains were typed by PFGE and plasmid profiling and investigated by molecular methods (PCR/sequencing) for the presence of several resistance genes, class 1 integrons and mutations in the quinolone resistance-determining regions. Plasmids conferring quinolone resistance were analysed by restriction fragment length polymorphism (RFLP) analysis, DNA hybridization, sequencing, replicon-typing PCR and mating experiments. All strains showed nalidixic acid resistance (MIC >or= 128 mg/L) together with a decreased susceptibility to ciprofloxacin (three strains with an MIC of 1 mg/L and six with an MIC of 0.25 mg/L), associated with mutations within the gyrA gene (Asp-87 --> Tyr-87). In three strains, qnrS1 genes were detected. Ampicillin resistance encoded by a bla(CTX-M3) gene and/or bla(TEM-1-like) gene was found in four strains. Three of these strains carried an approximately 45 kb conjugative plasmid, designated pRQ2006, harbouring qnrS1 and a Tn3-like transposon. Partial sequencing and RFLP of pRQ2006 indicated its similarity to the qnrS1 plasmid pAH03786 found in a Japanese Shigella flexneri 2b isolate. This is the first study describing the presence of qnrS1 genes in bacterial isolates from Turkey. The pRQ2006 plasmid seems to be more related to the S. flexneri 2b qnrS1 plasmid pAH0376 than to the Salmonella qnrS1-carrying plasmids pINF5 and TPqnrS-2.


The promoter activity of the phospholipase C-gamma2 gene is regulated by a cell-type-specific control element.
We have cloned and characterized a genomic DNA spanning the 5'-flanking region, the first and second exons, and the first intron of the human PLC-gamma2 gene. The proximal upstream region is highly GC-rich and lacks a TATA box, whereas the distal region contains several AT-rich tracts. Multiple transcription initiation sites were identified by primer extension analysis. Based on the transient transfection assays, the major transcriptional activation element was identified between -183 and +43 (G2SE) and a transcriptional repressive element was found between -303 and -184 (G2RE). The expression of PLC-gamma2 in various cell lines was examined using monoclonal anti-PLC-gamma2 antibody. PLC-gamma2 was highly expressed in B-cell lines such as Daudi, SP2, and Ramos cells, whereas it existed at very low levels in Jurkat, 3T3-L1, NBL-7, and C6Bu-1 cells. Moderate levels of PLC-gamma2 were also detected in C2C12, P19, U937, HL60, A431, and PC12 cells. The 4-kb genomic fragment upstream of -1,654 was able to activate transcription from the PLC-gamma2 promoter in Daudi and C2C12 cells, but not in Jurkat cells, which is consistent with the PLC-gamma2 protein expression levels in those cell lines. These results suggest that the cell-type-specific expression of PLC-gamma2 might be attributed to the transcriptional regulation by the upstream cis-element.


The accurate QSPR models to predict the bioconcentration factors of nonionic organic compounds based on the heuristic method and support vector machine.
The heuristic method (HM) and support vector machine (SVM) were used to build the linear and nonlinear quantitive structure-property relationship (QSPR) models for the prediction of the fish bioconcentration factors (BCF) for 122 diverse nonionic organic chemicals using the three descriptors calculated from the molecular structure alone and selected by HM. Both the linear and nonlinear model can give very satisfactory prediction results: the square of correlation coefficient R(2) was 0.929 and 0.953, the root mean square (RMS) error was 0.404 and 0.331, respectively for the whole dataset. The prediction result of the SVM model is better than that obtained by heuristic method, which proved SVM was a useful tool in the prediction of the BCF. At the same time, the HM model showed the influencing degree of different molecular descriptors on bioconcentration factors and then could improve the understanding for the bioconcentration mechanism of organic pollutants from molecular level.


Aspect Entropy Extraction Using Circular SAR Data and Scattering Anisotropy Analysis.
In conventional synthetic aperture radar (SAR) working modes, targets are assumed isotropic because the viewing angle is small. However, most man-made targets are anisotropic. Therefore, anisotropy should be considered when the viewing angle is large. From another perspective, anisotropy is also a useful feature. Circular SAR (CSAR) can detect the scattering variation under different azimuthal look angles by a 360-degree observation. Different targets usually have varying degrees of anisotropy, which aids in target discrimination. However, there is no effective method to quantify the degree of anisotropy. In this paper, aspect entropy is presented as a descriptor of the scattering anisotropy. The range of aspect entropy is from 0 to 1, which corresponds to anisotropic to isotropic. First, the method proposed extracts aspect entropy at the pixel level. Since the aspect entropy of pixels can discriminate isotropic and anisotropic scattering, the method prescreens the target from the isotropic clutters. Next, the method extracts aspect entropy at the target level. The aspect entropy of targets can discriminate between different types of targets. Then, the effect of noise on aspect entropy extraction is analyzed and a denoising method is proposed. The Gotcha public release dataset, an X-band circular SAR data, is used to validate the method and the discrimination capability of aspect entropy.


Interview with Thomas Powles: The use of durvalumab in urothelial cancer - the latest strides in immuno-oncology.
Thomas Powles speaks to Sebastian Dennis-Beron, Commissioning Editor: Dr. Thomas Powles is a clinical professor of genitourinary oncology and the lead for solid tumor research at Barts Cancer Institute in London. His main research interests are in genital and urinary cancers, in which he leads a spectrum of clinical studies from Phase I to randomized Phase III investigating novel targeted and immune therapies. His research also focuses on correlation of novel biomarkers and aims to define markers with prognostic value that may predict response or resistance to therapy. Dr. Powles is the author of more than 100 publications, on the editorial board of a number of journals, and part of the faculty for multiple global oncology meetings. He qualified from St Bartholomew's Medical School and received his MD from the University of London.


Watching others' actions: mirror representations in the parietal cortex.
An observation that neurons in the motor cortex of the monkey are active both when the monkey performs a specific action and when he watches an actor executing the same action led to the mirror-system hypothesis. This hypothesis suggests that primates perceive and interpret others' actions by generating an internal motor representation (e.g., simulation). Recent evidence suggests that humans have a similar mirror system. In this review, we focus on the essential congruence between the motor and visual properties of an action. We summarize behavioral and imaging studies in humans that show that observing others' actions can interfere with our own motor execution. We discuss a framework for understanding such an internal representation and suggest that the activity in the parietal cortex during observation of others' actions is based on the sensory-to-motor remapping properties of this region, which are necessary for fine control of our own actions.


Cardiovocal Syndrome: A rare cause of hoarseness in a patient with a history of pulmonary tuberculosis.
Hoarseness is a common clinical condition with underlying causes which can vary from reversible and benign to life-threatening and malignant. Cardiovocal syndrome may cause hoarseness secondary to left recurrent laryngeal nerve palsy when the recurrent laryngeal nerve is mechanically affected due to enlarged cardiovascular structures. We report a 28-year-old male who presented to the Government Medical College, Thiruvananthapuram, India, in 2013 with hoarseness. He had undergone irregular treatment for pulmonary tuberculosis (TB) two years previously. Fiber-optic laryngoscopy indicated left vocal cord palsy and a computed tomography scan of the chest revealed features of pulmonary hypertension with extensive enlargement of the pulmonary arteries. An echocardiogram confirmed severe pulmonary arterial hypertension with severe tricuspid regurgitation. He was diagnosed with left recurrent laryngeal palsy secondary to cardiovocal syndrome. Although reports exist of recurrent laryngeal palsy in TB, this case appears to be the first to report cardiovocal syndrome in a patient treated for pulmonary TB.


Invasion Rate Versus Diversity in Population Dynamics with Catastrophes.
A key question in the current diversity crisis is how diversity has been maintained throughout evolution and how to preserve it. Modern coexistence theories suggest that a high invasion rate of rare new types is directly related to diversity. We show that adding almost any mechanism of catastrophes to a stochastic birth, death, and mutation process with limited carrying capacity induces a novel phase transition characterized by a positive invasion rate but a low diversity. In this phase, new types emerge and grow rapidly, but the resulting growth of very large types decreases diversity. This model also resolves two major drawbacks of neutral evolution models: their failure to explain balancing selection without resorting to fitness differences and the unrealistic time required for the creation of the observed large types. We test this model on a classical case of genetic polymorphism: the HLA locus.


Reoxidation of the Thiol-Disulfide Oxidoreductase MdbA by a Bacterial Vitamin K Epoxide Reductase in the Biofilm-Forming Actinobacterium Actinomyces oris.
Posttranslocational protein folding in the Gram-positive biofilm-forming actinobacterium Actinomyces oris is mediated by a membrane-bound thiol-disulfide oxidoreductase named MdbA, which catalyzes oxidative folding of nascent polypeptides transported by the Sec translocon. Reoxidation of MdbA involves a bacterial vitamin K epoxide reductase (VKOR)-like protein that contains four cysteine residues, C93/C101 and C175/C178, with the latter forming a canonical CXXC thioredoxin-like motif; however, the mechanism of VKOR-mediated reoxidation of MdbA is not known. We present here a topological view of the A. oris membrane-spanning protein VKOR with these four exoplasmic cysteine residues that participate in MdbA reoxidation. Like deletion of the VKOR gene, alanine replacement of individual cysteine residues abrogated polymicrobial interactions and biofilm formation, concomitant with the failure to form adhesive pili on the bacterial surface. Intriguingly, the mutation of the cysteine at position 101 to alanine (C101A mutation) resulted in a high-molecular-weight complex that was positive for MdbA and VKOR by immunoblotting and was absent in other alanine substitution mutants and the C93A C101A double mutation and after treatment with the reducing agent β-mercaptoethanol. Consistent with this observation, affinity purification followed by immunoblotting confirmed this MdbA-VKOR complex in the C101A mutant. Furthermore, ectopic expression of the Mycobacterium tuberculosis VKOR analog in the A. oris VKOR deletion (ΔVKOR) mutant rescued its defects, in contrast to the expression of M. tuberculosis VKOR variants known to be nonfunctional in the disulfide relay that mediates reoxidation of the disulfide bond-forming catalyst DsbA in Escherichia coli Altogether, the results support a model of a disulfide relay, from its start with the pair C93/C101 to the C175-X-X-C178 motif, that is required for MdbA reoxidation and appears to be conserved in members of the class ActinobacteriaIMPORTANCE It has recently been shown in the high-GC Gram-positive bacteria (or Actinobacteria) Actinomyces oris and Corynebacterium diphtheriae that oxidative folding of nascent polypeptides transported by the Sec machinery is catalyzed by a membrane-anchored oxidoreductase named MdbA. In A. oris, reoxidation of MdbA requires a bacterial VKOR-like protein, and yet, how VKOR mediates MdbA reoxidation is unknown. We show here that the A. oris membrane-spanning protein VKOR employs two pairs of exoplasmic cysteine residues, including the canonical CXXC thioredoxinlike motif, to oxidize MdbA via a disulfide relay mechanism. This mechanism of disulfide relay is essential for pilus assembly, polymicrobial interactions, and biofilm formation and appears to be conserved in members of the class Actinobacteria, including Mycobacterium tuberculosis.


A Salmonella type three secretion effector/chaperone complex adopts a hexameric ring-like structure.
Many bacterial pathogens use type three secretion systems (T3SS) to inject virulence factors, named effectors, directly into the cytoplasm of target eukaryotic cells. Most of the T3SS components are conserved among plant and animal pathogens, suggesting a common mechanism of recognition and secretion of effectors. However, no common motif has yet been identified for effectors allowing T3SS recognition. In this work, we performed a biochemical and structural characterization of the Salmonella SopB/SigE chaperone/effector complex by small-angle X-ray scattering (SAXS). Our results showed that the SopB/SigE complex is assembled in dynamic homohexameric-ring-shaped structures with an internal tunnel. In this ring, the chaperone maintains a disordered N-terminal end of SopB molecules, in a good position to be reached and processed by the T3SS. This ring dimensionally fits the ring-organized molecules of the injectisome, including ATPase hexameric rings; this organization suggests that this structural feature is important for ATPase recognition by T3SS. Our work constitutes the first evidence of the oligomerization of an effector, analogous to the organization of the secretion machinery, obtained in solution. As effectors share neither sequence nor structural identity, the quaternary oligomeric structure could constitute a strategy evolved to promote the specificity and efficiency of T3SS recognition.


Toward selective functionalisation of oligosilanes: borane-catalysed dehydrogenative coupling of silanes with thiols.
Among established methods for transforming Si-H bonds, carbonyl hydrosilylation and heterodehydrogenative coupling with alcohols catalysed by B(C6F5)3 are shown to provide exceptionally clean routes to the derivatisation of tetra-substituted disilanes such as [Ph2SiH]2, giving no products resulting from Si-Si bond cleavage. Even higher activity is observed for the borane-catalysed dehydrogenative coupling of silanes with alkyl- and arylthiols, the first examples of such Si-S bond formation in the absence of a transition metal catalyst. Clean, quantitative syntheses of a range of thiosilanes are reported, and the lability of the Si-S linkage toward subsequent alcoholysis is investigated. The crystal structure of 2,3-disila-2,2,3,3-tetramethyl-1,4-benzodioxane is presented.


Michigan Alcoholism Screening Test to diagnose early alcoholism in a general practice.
General medical patients can be easily screened for alcohol dependence on a routine basis. The Michigan Alcoholism Screening Test (MAST) is a 24-item yes-no questionnaire concerning alcohol-related behavior. When it was included in the history self-administered to new patients in an internal medicine faculty-resident outpatient practice, it uncovered unsuspected alcohol dependence. The combined questionnaire (health history plus MAST) was given to 351 consecutive new outpatients. Thirteen (4%) were found to have positive MAST scores (greater than 7), indicating alcohol dependence. Nine patients were in the mild dependence range (7 to 20), two in the moderate range (20 to 30), and two in the severe range (30 to 53). All patients with significant MAST scores had complaints related to active drinking. The MAST score directed the physicians' therapeutic efforts in nine of 13 patients; in four the significant MAST score was ignored. The MAST is useful to detect unsuspected alcohol dependence in office practice. When used by faculty and residents trained in recognizing alcohol-related behavior, the test will properly direct therapeutic and rehabilitative efforts for those patients with the primary diagnosis of alcohol dependence.


The role of intracellular Na+ and mitochondria in buffering of kainate-induced intracellular free Ca2+ changes in rat forebrain neurones.
1. We have examined the mechanisms by which cultured central neurones from embryonic rat brain buffer intracellular Ca2+ loads following kainate receptor activation using fluorescent indicators of [Ca2+]i and [Na+]i. 2. Stimulation of cultured forebrain neurones with 100 microM kainate produced a rapid increase in [Ca2+]i that displayed a variable rate of recovery. Kainate also increased [Na+]i with a response that was slightly slower in onset and markedly slower in recovery. 3. The recovery of [Ca2+]i to baseline was not very sensitive to the [Na+]i. The magnitude of the increase in [Na+]i in response to kainate did not correlate well with the [Ca2+]i recovery time, and experimental manipulations that altered [Na+]i did not have a large impact on the rate of recovery of [Ca2+]i. 4. The recovery of [Ca2+]i to baseline was accelerated by the mitochondrial Na+-Ca2+ exchange inhibitor CGP-37157, suggesting that the recovery rate is influenced by release of Ca2+ from a mitochondrial pool and also that variation in the recovery rate is related to the extent of mitochondrial Ca2+ loading. Kainate did not alter the mitochondrial membrane potential. 5. These studies reveal that mitochondria have a central role in buffering neuronal [Ca2+]i changes mediated by non-N-methyl-D-aspartate (NMDA) glutamate receptors, and that the variation in recovery times following kainate receptor activation reflects a variable degree of mitochondrial Ca2+ loading. However, unlike NMDA receptor-mediated Ca2+ loads, kainate receptor activation has minimal effects on mitochondrial function.


PRScalc, a privacy-preserving calculation of raw polygenic risk scores from direct-to-consumer genomics data.
Currently, the Polygenic Score (PGS) Catalog curates over 400 publications on over 500 traits corresponding to over 3000 polygenic risk scores (PRSs). To assess the feasibility of privately calculating the underlying multivariate relative risk for individuals with consumer genomics data, we developed an in-browserPRS calculator for genomic data that does not circulate any data or engage in any computation outside of the user's personal device. A prototype personal risk score calculator, created for research purposes, was developed to demonstrate how the PGS Catalog can be privately and readily applied to readily available direct-to-consumer genetic testing services, such as 23andMe. No software download, installation, or configuration is needed. The PRS web calculator matches individual PGS catalog entries with an individual's 23andMe genome data composed of 600k to 1.4 M single-nucleotide polymorphisms (SNPs). Beta coefficients provide researchers with a convenient assessment of risk associated with matched SNPs. This in-browser application was tested in a variety of personal devices, including smartphones, establishing the feasibility of privately calculating personal risk scores with up to a few thousand reference genetic variations and from the full 23andMe SNP data file (compressed or not). The PRScalc web application is developed in JavaScript, HTML, and CSS and is available at GitHub repository (https://episphere.github.io/prs) under an MIT license. The datasets were derived from sources in the public domain: [PGS Catalog, Personal Genome Project].


Two cases of colon LST in transplanted liver patients.
Long-life immunosuppressive therapy increases the risk of de novo tumors in liver transplant recipients by decreasing the immune surveillance against malignant cells and oncogenic viruses. However, no cases of colon precancerous lesions have been reported in these subjects. Patient n. 1, a 73 yrs old male treated with calcineurin and purine synthesis inhibitors, showed at a per-protocol colono-scopy a 3 cm laterally spreading tumor (LST). Patient n. 2, a 73 yrs old male treated with calcineurin inhibitors, showed at a screening colonoscopy an LST occupying one third of the lumen circumference. Both subjects were asymptomatic, had been transplanted 14 years before, and in both cases, lesions showed severe dysplasia. LSTs represent 17.2% of advanced colorectal neoplasia (CRC) and risk factors are multifactorial. Immunosuppression may play a role which is however not completely understood. Based on this report, surveillance colonoscopy in liver transplanted patients should be considered.


[Psychotropic drugs and weight].
Psychotropic drugs can induce weight gain and a metabolic syndrome, reducing the quality of life and the life expectancy of patients. Weight gain is also responsible for poor adherence to treatment and therefore relapses of mental illness. This weight gain and the metabolic syndrome are insufficiently treated for obese patients with mental disorders. Recommendations exist.


Interaction of human chondrocytes and NIH/3T3 fibroblasts on chloric acid-treated biodegradable polymer surfaces.
It has been recognized that adhesion and proliferation of cells on biodegradable polymers such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and poly(lactide-co-glycolide) (PLGA) depend on the surface properties. The chloric acid (CA) treatment of these films was developed to increase surface wettability and to improve adhesion and proliferation of human chondrocytes and NIH/3T3 fibroblasts. The CA-treated films were characterized by the measurement of water contact angle, electron spectroscopy for chemical analysis (ESCA), and scanning electron microscopy (SEM). The changes of the film surface water contact angle gradually decreased with increase of CA treatment time, owing to the oxygen-based functional groups incorporated on the surface by CA treatment and were in the order PGA > PLGA > PLA due to the number of methyl group on the backbone chain. In ESCA analysis, as CA treatment time increased, the carbon (binding energy, approximately 285 eV) ratio decreased in film surfaces, whereas the oxygen (approximately 532 eV) ratio increased. The human chondrocytes from articular cartilage and mouse NIH/3T3 fibroblasts adhered for 1 day and grown for 2 days on the CA-treated films were counted and observed by SEM. As the surface wettability increased, the number of cells adhered and grown on the surface increased. In conclusion, this study demonstrated that the surface wettability of the biodegradable polymer plays an important role for cell adhesion and proliferation behavior for the application of the tissue engineering.


The mitochondrial retrograde signaling regulates Wnt signaling to promote tumorigenesis in colon cancer.
Cancer cells are known to upregulate aerobic glycolysis to promote growth, proliferation, and survival. However, the role of mitochondrial respiration in tumorigenesis remains elusive. Here we report that inhibition of mitochondrial function by silencing TFAM, a key transcription factor essential for mitochondrial DNA (mtDNA) replication and the transcription of mtDNA-encoded genes, markedly reduced tumor-initiating potential of colon cancer cells. Knockdown of TFAM significantly decreased mitochondrial respiration in colon cancer cells; however, the cellular levels of ATP remained largely unchanged as a result of increased glycolysis. This metabolic alteration rendered cancer cells highly susceptible to glucose deprivation. Interestingly, upregulation of glycolysis was independent of hypoxia-inducible factor-1 (HIF1) as TFAM knockdown cells fail to stabilize HIF1α under hypoxic conditions. Moreover, knockdown of TFAM results in decreased expression of genes-associated cancer stem cells downstream of Wnt/β-catenin signaling. Metabolic analysis reveals that the level of α-ketoglutarate (α-KG) was significantly upregulated in TFAM knockout cells. Silencing of prolyl hydroxylase domain-containing protein 2 (PHD2), a α-KG-dependent dioxyenase, rescued the expression of target genes of both HIF1α and Wnt/β-catenin. Furthermore, intestinal-specific knockout of TFAM prevents tumor formation in Apc-mutant mouse models of colon cancer. Taken together, our findings identify a novel role of mitochondria-mediated retrograde signaling in regulating Wnt signaling and tumor initiation in colon cancer.


Assessing and Identifying Improvements for Lung Cancer Screening in a Rural Population: A Human-Centered Design and Systems Approach.
Although lung cancer claims more lives than any other cancer in the United States, screening is severely underutilized, with <6% of eligible patients screened nationally in 2021 versus 76% for breast cancer and 67% for colorectal cancer. This article describes an effort to identify key reasons for the underutilization of lung cancer screening in a rural population and to develop interventions to address these barriers suitable for both a large health system and local community clinics. Data were generated from 26 stakeholder interviews (clinicians, clinical staff, and eligible patients), a review of key systems (Electronic Health Record and billing records), and feedback on the feasibility of several potential interventions by health care system staff. These data informed a human-centered design approach to identify possible interventions within a complex health care system by exposing gaps in care processes and electronic health record platforms that can lead patients to be overlooked for potentially life-saving screening. Deployed interventions included communication efforts focused on (1) increasing patient awareness, (2) improving physician patient identification, and (3) supporting patient management. Preliminary outcomes are discussed.


Incidence and ultrastructure of rudimentary cilia in benign and malignant peripheral nerve tumors.
The incidence and ultrastructure of cilia were examined in 26 peripheral nerve tumors. The cilia had the common features of elongated shafts and basal structures consisting of two basal bodies. The basal body contained two basal feet and some long striated rootlets. The cilia lacked central tubules and two dynein arms of subfiber A, suggesting that they were nonmotile and rudimentary. The quantitative results showed that the incidence of cilia was low in Schwann cells and fibroblasts of the 20 benign tumors and slightly higher in perineurial cells of the 12 neurofibromas. Conversely, the incidence was high in most of the 6 malignant tumors. Only one perineurial cell from normal mature nerve examined carried a cilium, although no cilia were seen in the other perineurial cells, Schwann cells, and fibroblasts observed. The results showed that ciliary formation was more marked in the more actively proliferating tissues such as malignant tumors, and less so in the less actively proliferating tissues such as benign tumors. Ciliary formation may be one of the ultrastructural features commonly observed in proliferative tissues.


Multi-scale Xception based depthwise separable convolution for single image super-resolution.
The main target of Single image super-resolution is to recover high-quality or high-resolution image from degraded version of low-quality or low-resolution image. Recently, deep learning-based approaches have achieved significant performance in image super-resolution tasks. However, existing approaches related with image super-resolution fail to use the features information of low-resolution images as well as do not recover the hierarchical features for the final reconstruction purpose. In this research work, we have proposed a new architecture inspired by ResNet and Xception networks, which enable a significant drop in the number of network parameters and improve the processing speed to obtain the SR results. We are compared our proposed algorithm with existing state-of-the-art algorithms and confirmed the great ability to construct HR images with fine, rich, and sharp texture details as well as edges. The experimental results validate that our proposed approach has robust performance compared to other popular techniques related to accuracy, speed, and visual quality.


Experience with guidelines for wastewater characterisation in The Netherlands.
In this paper experiences and results are presented with guidelines for wastewater characterisation that are standardised in The Netherlands for modelling purposes with ASM. The wastewater characterisation is based on a physical-chemical method to characterise the soluble and particulate fractions, combined with a BOD-analysis for characterising the biodegradable fraction of the influent COD. By following the guidelines, a sufficiently detailed and practical characterisation is obtained, and the results can be used for simulation studies on treatment plants for process optimisation, trouble-shooting and design assistance. At present, five years of experience is gained with the guidelines and they were used for the simulation of circa 100 treatment plants. The guidelines are evaluated as simple and easy to implement in routine analysis programs.


Substituted 1,2,4-Triazoles as Novel and Selective Inhibitors of Leukotriene Biosynthesis Targeting 5-Lipoxygenase-Activating Protein.
5-Lipoxygenase-activating protein (FLAP) is a regulator of cellular leukotriene biosynthesis, which governs the transfer of arachidonic acid (AA) to 5-lipoxygenase for efficient metabolism. Here, the synthesis and FLAP-antagonistic potential of fast synthetically accessible 1,2,4-triazole derivatives based on a previously discovered virtual screening hit compound is described. Our findings reveal that simple structural variations on 4,5-diaryl moieties and the 3-thioether side chain of the 1,2,4-triazole scaffold markedly influence the inhibitory potential, highlighting the significant chemical features necessary for FLAP antagonism. Comprehensive metabololipidomics analysis in activated FLAP-expressing human innate immune cells and human whole blood showed that the most potent analogue 6x selectively suppressed leukotriene B4 formation evoked by bacterial exotoxins without affecting other branches of the AA pathway. Taken together, the 1,2,4-triazole scaffold is a novel chemical platform for the development of more potent FLAP antagonists, which warrants further exploration for their potential as a new class of anti-inflammatory agents.


In vitro oxidative degradation of a spinal posterior dynamic stabilization device.
This study quantified the changes of the frequency-dependant viscoelastic properties of the BDyn (S14 Implants, Pessac, France) spinal posterior dynamic stabilization (PDS) device due to in vitro oxidation. Six polycarbonate urethane (PCU) rings and six silicone cushions were degraded using a 20% hydrogen peroxide/0.1 M cobalt (II) chloride hexahydrate, at 37°C, for 24 days. The viscoelastic properties of the individual components and the components assembled into the BDyn PDS device were determined using Dynamic Mechanical Analysis at frequencies from 0.01 to 30 Hz. Attenuated Total Reflectance Fourier Transform Infra-Red spectra demonstrated chemical structure changes, of the PCU, associated with oxidation while Scanning Electron Microscope images revealed surface pitting. No chemical structure or surface morphology changes were observed for the silicone cushion. The BDyn device storage and loss stiffness ranged between 84.46 N/mm to 99.36 N/mm and 8.13 N/mm to 21.99 N/mm, respectively. The storage and loss stiffness for the components and BDyn device increased logarithmically with respect to frequency. Viscoelastic properties, between normal and degraded components, were significantly different for specific frequencies only. This study demonstrates the importance of analyzing changes of viscoelastic properties of degraded biomaterials and medical devices into which they are incorporated, using a frequency sweep. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1237-1244, 2018.


Tunable Orthogonal Reversible Covalent (TORC) Bonds: Dynamic Chemical Control over Molecular Assembly.
Dynamic assembly of macromolecules in biological systems is one of the fundamental processes that facilitates life. Although such assembly most commonly uses noncovalent interactions, a set of dynamic reactions involving reversible covalent bonding is actively being exploited for the design of functional materials, bottom-up assembly, and molecular machines. This Minireview highlights recent implementations and advancements in the area of tunable orthogonal reversible covalent (TORC) bonds for these purposes, and provides an outlook for their expansion, including the development of synthetically encoded polynucleotide mimics.


Mechanism for correlation in a coherent electron beam.
The correlation of electron counts from two detectors illuminated by a coherent electron beam is analyzed by associating the path of the electron beam through the lenses with the direct Coulomb interaction between two individual electrons. This is shown to lead to a full statistical description of the electron counts. The dominant contribution to the correlation is found to be due to the trajectory displacement caused by the repulsive Coulomb interaction between the first anode and the cathode tip, and the correlation of electron counts is found to depend on the amount of defocusing on the shift of the virtual source for two electrons within the correlation time. The Coulomb scatterings, which altered the direction of two neighbor electrons, occur during the acceleration, leading to a significant decrease in the electron density. The Coulomb potential with no screening will then cause large-angle scattering of nearest-neighbor electrons within the correlation time. These results are consistent with those obtained by a previous experiment.


Synthesis of cyclopentano-N-methylphosphatidylethanolamines: aminolysis during the use of methylamine.
Monomethylamine and N-benzyl-N-methylamine were used as nucleophiles in the amination of the bromoethylester of cyclopentano-phosphatidic acid. The former reagent led to extensive aminolysis and the quantitative formation of N-methylpalmitamide, rather than the desired cyclopentano-phosphatidyl-N-methylethanolamine. However, the method of Shapiro and Rabinsohn (1964. Biochemistry. 3:603-605), in which N-benzyl-N-methylamine was used as a nucleophile, was adapted for a successful synthesis.


Quasi-phase-matched multispectral Kerr frequency comb.
We study a new type of Kerr frequency comb where the momentum conservation law is fulfilled by azimuthal modulation of the waveguide dispersion. The concept can expand the parametric range in which a Kerr frequency comb is obtained. In a good agreement with the theoretical analysis, we demonstrate a multispectral Kerr frequency comb covering important fiber-optic communication bands. Comb coherence and absence of a sub-comb offset are confirmed by continuous-wave heterodyne beat note and amplitude noise spectra measurements. The device can be used for achieving broadband optical frequency synthesizers and high-capacity coherent communication.


Voluntary self-amputation of the colon.
We describe an unusual case of self-amputation of a transanal prolapsed colorectal segment by a 48-year-old mentally impaired woman. The surgical procedure and favourable outcome are reported.


Opioid Use Initiation, Progression, and Motivations Among OEF/OIF/OND-Era Veterans in New York City: An Age-Period-Cohort Analysis.
This study examines the temporal relationship between prescription opioid (PO) and heroin use among veterans in New York City. Drawing on survey data from a convenience sample of 214 Operation Enduring Freedom/Operation Iraqi Freedom/Operation New Dawn (OEF/OIF/OND)-era opioid-using military veterans, analyses demonstrate substantial cohort-level variation. Most notably, heroin use prior to PO initiation and prior to military enlistment was reported more frequently among the cohorts born prior to 1970 and after 1984. Across all cohorts, high percentages of participants reported alleviation of emotional/psychological pain as a reason for both PO and heroin use. Cohort-level variation highlights the need for tailored interventions and targeted prevention efforts.


Florida Red Tide: Inhalation Toxicity of Karenia brevis Extract in Rats.
Brevetoxins are neurotoxins produced by the marine dinoflagellate Karenia brevis. Histopathologic examination of marine mammals dying following repeated exposure of brevetoxins during red tide events suggests that the respiratory tract, nervous, hematopoietic, and immune systems are potential targets for toxicity in repeatedly exposed individuals. The purpose of this experiment was to evaluate the effects of repeated inhalation of K. brevis extract on these potential target systems in rats. Male Sprague-Dawley rats were exposed four hours/day, five days/week for up to four weeks to target concentrations of 200 and 1000 μg/L K. brevis extract (approximately 50 and 200 μg/L brevetoxin-like compounds; positive neurotoxicity in a fish bioassay). Control rats were sham exposed to air. Immunohistochemical staining of pulmonary macrophages indicated deposition of brevetoxin-like compound within the lung. However, exposure resulted in no clinical signs of toxicity or behavioral changes. There were no adverse effects on hematology or serum chemistry. No histopathological changes were observed in the nose, lung, liver, kidneys, lymph nodes, spleen, or brain of exposed rats. Immune suppression was suggested by reduced responses of spleen cells in the IgM-specific antibody-forming plaque cell response assay and reduced responses of lymphocytes to mitogen stimulation in vitro. Differences between responses observed in rats in this study and those observed in manatees may be a function of dose or species differences in sensitivity.


Trends in Tobacco Product Use Patterns Among U.S. Youth, 1999-2014.
We examined trends in seven mutually exclusive tobacco product use patterns (T-PUPs) in nationally representative samples of U.S. youth over time and age. We used time varying effect modeling on National Youth Tobacco Surveys, 1999-2014 (N = 38662, 9-17 years, M = 15.02). Regression coefficients were estimated as a non-parametric function of time. T-PUPs were cigarette only, non-cigarette combustible only, noncombustible only, non-cigarette combustible and noncombustible dual, cigarette and noncombustible dual, cigarette and non-cigarette combustible dual, and POLY (i.e. cigarettes, non-cigarette combustibles, and noncombustibles) use. Among youth tobacco users, cigarette only use was the predominant T-PUP from 1999 to 2010. After 2010 and 2013, non-cigarette combustible only (AOR 1.38, CI = 1.02-1.87) and noncombustible only (AOR 1.57, CI = 1.00-2.45) use became more prevalent than cigarette only use. In 2011, dual and POLY T-PUPs were on the rise although not significantly different from cigarette only use. Cigarette only use was the predominant T-PUP among 11- to 17-year-old tobacco users. Non-cigarette combustible only (AOR 0.14, CI = 0.10-0.19), noncombustible only (AOR 0.01, CI = 0.008-0.02), non-cigarette combustible and noncombustible (AOR 0.01, CI = 0.01-0.03), cigarette and noncombustible (AOR 0.02, CI = 0.01-0.04), cigarette and non-cigarette combustible (AOR 0.32, CI = 0.24-0.43), and POLY (AOR 0.02, CI = 0.01-0.04) use were less prevalent than cigarette only use at age 17. Non-cigarette, dual, and POLY T-PUPs are rising among youth tobacco users. Screening for all tobacco use and delivering treatment during pediatrician visits should be standard clinical practice. Tracking trends in tobacco product use patterns (T-PUPs) over time and age is necessary to achieve Healthy People 2020 goal of reducing tobacco use among youth. Trends over time show a rise of non-cigarette T-PUPs especially noncombustible products but cigarette only use remains the most prevalent among 11- to 17-year-old tobacco users. The recent extension of FDA's regulatory jurisdiction over all tobacco products (e.g. e-cigarettes, hookah) is a step toward comprehensive tobacco control especially among youth. Public health practitioners should extend prevention and cessation efforts among youth to T-PUPs beyond exclusive cigarette smoking.


Misreport of energy intake assessed with food records and 24-h recalls compared with total energy expenditure estimated with DLW.
The accuracy of dietary assessment methods has rarely been validated using precise techniques. The objective of this work was to evaluate the validity of energy intake (EI) estimated with food records (FRs) and 24-h recalls (24hRs) against total energy expenditure (EE) estimated by the doubly labeled water (DLW) method. In addition, the magnitude of EI under-reporting was assessed along with its associated characteristics. The studied group included 83 adults between 20 and 60 years of age who were recruited from a population-based sample. Within-person variation-adjusted means of EI estimated from two FRs and three 24hRs were compared with EE estimated using the DLW method multiple-point protocol. The Wilcoxon signed-rank test was used to assess the differences between EI and EE, whereas Bland-Altman and survival-agreement plots assessed the agreement between the estimates. The mean EE (2540 kcal) was greater than the mean reported EI for both dietary assessment methods (FR: 1774 kcal; 24hR: 1658 kcal, P<0.01). The frequency of under-reporting was lower (20%) for EI estimated with the 24hR than that estimated with the FR (32%). Men presented lower magnitude of under-reported EI than women did. For women, differences between EI and EE were lower with FR than with 24hR. Overall, FR and 24hR showed similar performance. The mean under-reported EI was ~30% for both methods. Irregular meal habits, smoking and low education were associated with the under-report of EI. Both FR and 24hR are subjected to bias suggesting the need of refining the procedures applied in dietary assessment methods.


Prevalence of abdominal obesity in non-obese adolescents: a North Indian adolescent study.
Context Childhood obesity is defined based on body mass index (BMI) criteria. Asian adults have more abdominal adiposity as compared to Western people for an equivalent BMI. Similarly, children who are non-obese/overweight by BMI criteria may have abdominal obesity. It is important to identify and target this population to reduce future cardiovascular risk. Objective To evaluate and inter-relate the prevalence of obesity subtypes (generalized and abdominal) in school going adolescents. Methods A randomized cross-sectional community based study was conducted, which was carried out in 13 schools of Chandigarh, a North Indian city. 10,037 school going adolescents of age 10-18 years were evaluated. Results The prevalence of overweight and generalized obesity in present study was 9.3 and 4.9% respectively. Generalized obesity was observed in 4.2% of males and 5.7% of females (p-Value = 0.001). The abdominal obesity was noted in 5.4% of children and its prevalence increased progressively with age. The prevalence was highest in 16-18 years age-group (7.6%, p<0.001). 39.3% of abdominally obese subjects were not obese by BMI criteria, while prevalence of abdominal obesity in nonobese adolescents was 2.24%. It was more prevalent in females (3.7%) than males (3%, p=0.025). Prevalence of obesity was 7.9 and 1.8% in private and government schools respectively (p<0.001). Conclusions Abdominal obesity is more prevalent than generalized obesity and shows increasing trend with age. Interestingly, over one third of centrally obese adolescents are not obese by BMI criteria. This underlines the importance of waist circumference measurement in addition to BMI while assessing obesity.


Rib cartilage grafts for the treatment of posterior glottic and subglottic stenosis in children.
Posterior glottic and subglottic stenosis from endotracheal intubation in children can be managed endoscopically with varying success. Open surgical treatment offers a better potential for correction with a single procedure in moderate and severe cases. The open method consists of splitting the scar and cricoid cartilage posteriorly to the level of the interarytenoid muscle, then stenting the incised cricoid open with a rib cartilage graft. Use of this method is described, and results in 12 cases are reported. Decannulation was achieved in ten patients. In all patients who were decannulated, good exercise tolerance, freedom from aspiration, and an adequate voice quality were achieved.


Sharing of Alcohol-Related Content on Social Networking Sites: Frequency, Content, and Correlates.
The present study aimed to explore students' reports of their sharing of alcohol-related content on different social networking sites (i.e., frequency of sharing and connotations of alcohol-related posts), and to identify indicators of such posting. Students at the four largest institutions for higher education in Bergen, Norway, were invited to participate in an Internet-based survey. The sample size was 11,236 (a 39.4% response rate). The survey included questions about disclosure of alcohol-related content on social networking sites, alcohol use (using the Alcohol Use Disorders Identification Test), personality factors (using the Mini-IPIP), and demographic characteristics. Binary logistic regressions were used to analyze indicators of frequent sharing of alcohol-related content depicting positive and negative aspects of alcohol use. A majority of the students had posted alcohol-related content (71.0%), although few reported having done so frequently. Positive aspects of alcohol use (e.g., enjoyment or social community) were most frequently shared. Young, single, and extroverted students with high alcohol consumption were more likely to report frequent sharing of alcohol-related content. Positive attitudes toward posting alcohol-related content and reports of exposure to such content particularly increased the likelihood of one's own posting of alcohol-related content. Positive aspects of alcohol use seem to be emphasized on social networking sites. Sharing of alcohol-related content is associated with heightened alcohol use, which implies that such sites can be relevant for prevention agents. Social influence from social networking sites, such as exposure to others' alcohol-related content, is associated with one's own sharing of similar content.


Protein kinase D at the Golgi controls NLRP3 inflammasome activation.
The inflammasomes are multiprotein complexes sensing tissue damage and infectious agents to initiate innate immune responses. Different inflammasomes containing distinct sensor molecules exist. The NLRP3 inflammasome is unique as it detects a variety of danger signals. It has been reported that NLRP3 is recruited to mitochondria-associated endoplasmic reticulum membranes (MAMs) and is activated by MAM-derived effectors. Here, we show that in response to inflammasome activators, MAMs localize adjacent to Golgi membranes. Diacylglycerol (DAG) at the Golgi rapidly increases, recruiting protein kinase D (PKD), a key effector of DAG. Upon PKD inactivation, self-oligomerized NLRP3 is retained at MAMs adjacent to Golgi, blocking assembly of the active inflammasome. Importantly, phosphorylation of NLRP3 by PKD at the Golgi is sufficient to release NLRP3 from MAMs, resulting in assembly of the active inflammasome. Moreover, PKD inhibition prevents inflammasome autoactivation in peripheral blood mononuclear cells from patients carrying NLRP3 mutations. Hence, Golgi-mediated PKD signaling is required and sufficient for NLRP3 inflammasome activation.


Complexity of the major surface protease (msp) gene organization in Leishmania (Viannia) braziliensis: evolutionary and functional implications.
The major surface protease (msp or gp63) of Leishmania plays a major role in the host-parasite interaction. We analysed here the structure of the msp gene locus in Leishmania (Viannia) braziliensis and compared it to results obtained in other species. Physical mapping of cosmid contigs revealed a minimum of 37 genes per haploid genome and at least 8 different msp gene families. Within the same organism, these genes showed a nucleotide sequence varying in certain stretches from 3 to 34%, and a mosaic structure. From an evolutionary point of view, major differences were observed between subgenera Viannia and Leishmania, both in terms of msp gene number and sequence. Within subgenus Viannia, phenetic analysis revealed three clusters in which sequence variants of L. (Viannia) braziliensis and L. (Viannia) guyanensis were interspersed. Functional implications of our results were explored from predicted L. (Viannia) braziliensis protein sequences: regions encoding the msp catalytic site showed a conserved sequence, while regions encoding surface domains possibly involved in the host-parasite interaction (macrophage adhesion sites and immunodominant B-cell and T-cell epitopes) were variable. We speculate that this would be an adaptive strategy of the parasite.


Ferritin light chain promotes the reprogramming of glioma immune microenvironment and facilitates glioma progression.
Background: Tumor-associated macrophages (TAMs), the most abundant non-tumor cell population in the glioma microenvironment, play a crucial role in immune evasion and immunotherapy resistance of glioblastoma (GBM). However, the regulatory mechanism of the immunosuppressive TME of GBM remains unclear. Methods: Bioinformatics were used to analyse the potential role of ferritin light chain (FTL) in GBM immunology and explore the effects of FTL on the reprogramming of the GBM immune microenvironment and GBM progression. Results: The FTL gene was found to be upregulated in TAMs of GBM at both the bulk and single-cell RNA-seq levels. FTL contributed to the protumor microenvironment by promoting M2 polarization in TAMs via inhibiting the expression of iPLA2β to facilitate the ferroptosis pathway. Inhibition of FTL in TAMs attenuated glioma angiogenesis, promoted the recruitment of T cells and sensitized glioma to anti-PD1 therapy. Conclusion: Our study suggested that FTL promoted the development of an immunosuppressive TME by inducing M2 polarization in TAMs, and inhibition of FTL in TAMs reprogrammed the TME and sensitized glioma to anti-PD1 therapy, providing a new strategy for improving the therapeutic effect of anti-PD1.


Comparative genomics on Vangl1 and Vangl2 genes.
WNT signals are transduced to the beta-catenin pathway or the planar cell polarity (PCP) pathway. WNT - beta-catenin pathway is implicated in carcinogenesis, while WNT-PCP pathway is implicated in cell motility and metastasis. Drosophila Van Gogh (Vang), Frizzled (Fz), Starry night (Stan), Prickle (Pk) and Diego (Dgo) are PCP signaling molecules. Vangl1 (Strabismus 2) and Vangl2 (Strabismus 1 or Ltap) are mammalian homologs of Drosophila Vang interacting with PRICKLE1, PRICKLE2, ANKRD6, DVL1, DVL2, DVL3, KAI1 and MAGI3. Here we identified and characterized rat Vangl1 and Vangl2 genes by using bioinformatics. Rat Vangl1 gene, consisting of eight exons, was located within AC098913.7 and AC108524.6 genome sequences. Rat Vangl2 gene, consisting of eight exons, was located within AC118856.3 and AC115243.5 genome sequences. Exon-intron structure of mammalian Vangl1 and Vangl2 orthologs was well conserved. E47 and double ELK1-binding sites were conserved among promoters of mammalian Vangl1 orthologs. PAX4, NFkappaB, HNF4, SOX9, RFX1, and POU2F1 (OCT1)-binding sites were conserved among promoters of mammalian Vangl2 orthologs. Rat Vangl1 (526 aa) and Vangl2 (521 aa) were four-transmembrane proteins with 71.5% total-amino-acid identity. Ser cluster motif (SxxSxxSxxSxxSxxS) in the N-terminal cytoplasmic region and PDZ-binding motif in the C-terminal cytoplasmic tail were evolutionarily conserved among vertebrate Vangl1 and Vangl2 orthologs. This is the first report on rat Vangl1 and Vangl2 genes as well as on comparative genomics for Vangl1 and Vangl2 orthologs.


Mediastinal mass after coronary artery bypass surgery with internal mammary artery grafts.
The usual appearance of the postoperative chest radiograph in patients after coronary artery bypass surgery is well known. The article describes an unusual case with development of a right anterior mediastinal mass after bypass surgery with internal mammary artery grafts. This mediastinal mass, initially detected on a chest radiograph, was confirmed by contrast-enhanced chest CT to be intrathoracic fat from the right internal mammary artery pedicle.


Seasonal variation of natural mortality factors of the guava psyllid Triozoida limbata.
It is important to understand how components of the agroecosystem interfere with the attack of a pest species and their seasonality in order to use these components in IPM programs. This study focused on the evaluation of the seasonality of natural control factors associated with the guava psyllid Triozoida limbata (Enderlein) in Brazil. Life-table data were collected from an experimental guava orchard during four periods that roughly represented four seasons. Natural mortality was monitored daily through the immature stages, and the relative importance of each natural mortality factor and its seasonality was determined. Significant statistical differences were observed in the mortality during the four periods (P < 0.05). Several factors contributed to the mortality of T. limbata, including rainfall, physiological disturbance, the parasitoid Psyllaephagus sp. (Hymenoptera: Encyrtidae) and specific predators: syrphids, predatory wasps and other generalist predators. Depending on the location of nymphs (exposed or inside galls), the relative importance of the different natural mortality factors changed. The principal component analysis (PCA) showed some trends in the relationship of natural control agents and weather conditions. For example, the occurrence of predatory wasps was positively correlated with temperature and occurrence of winds; the occurrence of syrphids and Psyllaephagus sp. were negatively correlated with temperature and winds; and the occurrence of other generalist predators were negatively correlated with the occurrence of rainfall and photoperiod. The results showed the importance of natural mortality factors for the management of T. limbata and their changes through the different seasons which should be considered when implementing IPM programs in guava orchards.


The distinct functions of two classical arabinogalactan proteins BcMF8 and BcMF18 during pollen wall development in Brassica campestris.
Arabinogalactan proteins (AGPs) are extensively glycosylated hydroxyproline-rich glycoproteins ubiquitous in all plant tissues and cells. AtAGP6 and AtAGP11, the only two functionally known pollen-specific classical AGP encoding genes in Arabidopsis, are reported to have redundant functions in microspore development. BcMF18 and BcMF8 isolated from Brassica campestris are the orthologues of AtAGP6 and AtAGP11, respectively. In contrast to the functional redundancy of AtAGP6 and AtAGP11, single-gene disruption of BcMF8 led to deformed pollen grains with abnormal intine development and ectopic aperture formation in B. campestris. Here, we further explored the action of BcMF18 and its relationship with BcMF8. BcMF18 was specifically expressed in pollen during the late stages of microspore development. Antisense RNA transgenic lines with BcMF18 reduction resulted in aberrant pollen grains with abnormal cellulose distribution, lacking intine, cytoplasm and nuclei. Transgenic plants with repressive expression of both BcMF8 and BcMF18 showed a hybrid phenotype, expressing a mixture of the phenotypes of the single gene knockdown plant lines. In addition, we identified functional diversity between BcMF18/BcMF8 and AtAGP6/AtAGP11, mainly reflected by the specific contribution of BcMF18 and BcMF8 to pollen wall formation. These results suggest that, unlike the orthologous genes AtAGP6 and AtAGP11 in Arabidopsis, BcMF18 and BcMF8 are both integral to pollen biogenesis in B. campestris, acting through independent pathways during microspore development.


Investigation of Controlled Growth of Metal-Organic Frameworks on Anisotropic Virus Particles.
Biomimetic mineralization with metal-organic frameworks (MOF), typically zeolitic imidazolate framework-8 (ZIF-8), is an emerging strategy to protect sensitive biological substances against denaturing environmental stressors such as heat and proteolytic agents. Additionally, this same biomimetic mineralization process has the potential of being used to create distinct core-shell architectures using genetically or chemically modified viral nanoparticles. Despite the proliferation of examples for ZIF-8 growth on biological or proteinaceous substrates, systematic studies of these processes are few and far between. Herein, we employed the tobacco mosaic virus (TMV) as a model biological template to investigate the biomimetic mineralization of ZIF-8, which has been proven to be a robust MOF for encasing and protecting inlaid biological substances. Our study shows a systematic dependence upon ZIF-8 crystallization parameters, e.g., ligand to metal molar ratio and metal concentration, which can yield several distinct morphologies of TMV@ZIF-8 composites and phases of ZIF-8. Further investigation using charged synthetic conjugates, time dependent growth analysis, and calorimetric analysis has shown that the TMV-Zn interaction plays a pivotal role in the final morphology of the TMV@ZIF-8, which can take the form of either core-shell bionanoparticles or large crystals of ZIF-8 with entrapped TMV located exclusively on the outer facets. The design rules outlined here, it is hoped, will provide guidance in biomimetic mineralization of MOFs on proteinaceous materials using ZIF-8.


Quality of care in breast cancer centers: results of benchmarking by the German Cancer Society and German Society for Breast Diseases.
A total of 218 breast cancer centers, with 274 operating sites, have been certified since 2003 in accordance with the criteria set out by the German Cancer Society (Deutsche Krebsgesellschaft) and the German Society for Breast Diseases (Deutsche Gesellschaft für Senologie). Most of these centers are located in Germany, but centers in Austria, Switzerland, and Italy have also been certified. This paper presents the results for quality indicators (QIs) from 2009 to 2012, based on data from 195,342 primary breast cancer patients, in order to illustrate the development and progress of these centers. Descriptive results on interdisciplinary collaboration (6 QIs), guideline adherence (15 QIs), and specialist expertise (7 QIs) are reported over time. Fulfillment of the certification requirements was high and remained relatively stable over time, with the extent of variation between sites declining. Sites that do not reach the target values are asked to comment on the discrepancies, and their comments, as well as the QI results, are used by centers and auditors during an on-site auditing process and to constantly revise the QIs set out by the guideline and the certification commission.


Incorporating data from various trial designs into a mixed treatment comparison model.
Estimates of relative efficacy between alternative treatments are crucial for decision making in health care. Bayesian mixed treatment comparison models provide a powerful methodology to obtain such estimates when head-to-head evidence is not available or insufficient. In recent years, this methodology has become widely accepted and applied in economic modelling of healthcare interventions. Most evaluations only consider evidence from randomized controlled trials, while information from other trial designs is ignored. In this paper, we propose three alternative methods of combining data from different trial designs in a mixed treatment comparison model. Naive pooling is the simplest approach and does not differentiate between-trial designs. Utilizing observational data as prior information allows adjusting for bias due to trial design. The most flexible technique is a three-level hierarchical model. Such a model allows for bias adjustment while also accounting for heterogeneity between-trial designs. These techniques are illustrated using an application in rheumatoid arthritis.


Effect of chemomechanical caries removal on bonding of self-etching adhesives to caries-affected dentin.
To evaluate the effect of enzyme-based (Papacárie) and sodium-hypochlorite-based (Carisolv) chemomechanical caries removal methods on bonding of self-etching adhesives to caries-affected dentin, in comparison to the standard rotary-instrument caries removal method. Seventy-eight carious permanent molars exhibiting frank cavitation into dentin were used. Forty-eight teeth were randomly divided into three groups, according to the caries excavation methods: (i) Papacárie, (ii) Carisolv and (iii) a round steel bur. After caries removal, each group was subdivided into two groups for two-step (Clearfil SE Bond) or one-step (Clearfil S3 Bond) self-etching adhesive application and resin composite buildups. Bonded specimens were sectioned into beams for microtensile bond strength testing. Bond strength data were analyzed using three-way ANOVA and Tukey's test. For interfacial nanoleakage evaluation using a field-emission scanning electron microscope, caries was similarly removed from the remaining thirty carious molars, bonding was performed as for bond strength testing, and the teeth were sectioned. RESULTS of three-way ANOVA revealed that bond strength was significantly affected by "adhesive" (p<0.001) and "dentin" (p<0.001), but not "caries excavation methods" (p>0.05). The bond strength of the two-step self-etching adhesive was significantly higher than that of the one-step self-etching adhesive (p<0.001). Conversely, the bond strength of self-etching adhesives to sound dentin was significantly higher than to residual caries-affected dentin (p<0.001). Greater silver penetration was observed in the bonded interfaces of residual caries-affected dentin and in interfaces bonded with the one-step self-etching adhesive vs those bonded with the two-step self-etching adhesive. Chemomechanical caries removal did not affect the bonding of self-etching adhesives to caries-affected dentin as compared to caries excavation with rotary instruments.


Dreaming of a learning task is associated with enhanced memory consolidation: Replication in an overnight sleep study.
Sleep following learning benefits memory. One model attributes this effect to the iterative "reactivation" of memory traces in the sleeping brain, demonstrated in animal models. Although technical limitations prohibit using the same methods to observe memory reactivation in the human brain, the study of mental activity during sleep provides an alternative method of observing memory activation during sleep. In fact, the content of dream experience may reflect the process of memory reactivation and consolidation in the sleeping brain. In line with this hypothesis, we previously reported that dreaming about a spatial learning task during a nap strongly predicts subsequent performance improvements. Here, we replicate this observation in an overnight sleep study, for the first time demonstrating that pre-sleep training on a virtual maze navigation task is reflected in dreams reported from all phases of sleep, with unambiguous representation of the task in dream content associated with improved next-morning performance. These observations are consistent with reactivation-based models of memory consolidation in sleep, confirming our earlier finding that the cognitive-level activation of recent experience during sleep is associated with subsequent performance gains.


EVI1 upregulates PTGS1 (COX1) and decreases the action of tyrosine kinase inhibitors (TKIs) in chronic myeloid leukemia cells.
Tyrosine kinase inhibitors (TKIs) are highly effective in treating chronic myelogenous leukemia (CML). However, primary and acquired drug resistance to TKIs have been reported. In this study, we used RNA sequencing followed by RQ-PCR to show that the proto-oncogene EVI1 targets the drug-metabolizing gene PTGS1 in CML. The PTGS1 promoter element had an EVI1 binding site, and CHIP assay confirmed its presence. Data from a publicly available CML microarray dataset and an independent set of CML samples showed a significant positive correlation between EVI1 and PTGS1 expression in CML. Downregulation of EVI1 in K562 cells and subsequent treatment with TKIs resulted in a lower IC50 in the control cells. Furthermore, combined inhibition of BCR-ABL with imatinib and PTGS1 with FR122047 (PTGS1 inhibitor) synergistically reduced the viability of imatinib-resistant K562 cells. We conclude that elevated EVI1 expression contributes to TKIs resistance and that combined inhibition of PTGS1 and BCR-ABL may represent a novel therapeutic approach.


Three-dimensional computerized reconstruction of histologic serial sections for orthopedic research.
New programs in image processing and computer graphics can now render three-dimensional images from serial tissue slices. Although neuroanatomists have used these techniques most intensively, there are exciting applications of this methodology in orthopedic research and investigative embryology. Three-dimensional reconstructions of histologic sections from newborn clubfeet have been used to determine the rotational alignment of the hindfoot bones. Similar studies could be undertaken to improve understanding of the normal and pathologic perinatal development of the spine, hip, and hand.


[Radiotherapy and chemotherapy for esophageal cancer].
The nonsurgical gold standard treatment for esophageal cancer is radiotherapy, but chemoradiation therapy using anticancer agents concurrently is becoming standard management. With chemoradiation adverse responses in the acute stage may be enhanced in comparison with conventional radiation therapy alone, but improved treatment outcomes are reported. A consensus has nearly been reached on the standard radiation technique and dose, and a guideline changes, but drug type, timing, and optimum dosage in chemoradiation are unclear. Further future study is thus necessary. In this report, recent trends in radiotherapy and chemoradiation are outlined and future problems described.


Molecular heterogeneity at the phenylalanine hydroxylase locus in the population of the south-west of England.
The phenylalanine hydroxylase gene locus has been studied in 35 independent phenylketonuric families in the south-west of England using RFLP haplotype patterns and allele specific oligonucleotide probes. Haplotype 3 was the most common pattern on mutant chromosomes and there was strict linkage disequilibrium between this haplotype and the splice mutation in exon 12. The R408W mutation in exon 12 occurred on both haplotypes 1 and 2. The R126Q mutation in exon 7 was found only on a rare haplotype 28 pattern. No gene carried the R158Q mutation. More than 60% of mutant genes did not carry these four mutations which were originally described in other European populations. We suggest that the splice mutation arose as a single event and spread throughout northern Europe by population migration and admixture. In addition, we believe the haplotype/mutation associations seen in our population are a reflection of the mixed ancestry of the inhabitants of the British Isles.


Three new species of the cutthroat eel genus Dysomma, with comments on the variation of D. taiwanense (Anguilliformes: Synaphobranchidae).
Three new species of eels of the genus Dysomma are described from the western North Pacific Ocean off Taiwan. Dysomma formosa sp. nov., described from 34 specimens, differs from the congeners in having single row of 11-14 large compound teeth followed by 3-10 smaller teeth on lower jaw, 17-33 lateral-line pores, and 128-133 total vertebrae. Dysomma brachygnathos sp. nov., described from two specimens, differs from the congeners in the lack of a pectoral fin, having an anterior position of anus (preanal length 17.8-ca. 18.6% SL), 23-32 lateral line pores, a short lower jaw, and 131+‒136 total vertebrae. Dysomma robinsorum sp. nov., described from four specimens, differs from the congeners in having an anterior position of anus; preanal length 15.5-16.9% SL, no intermaxillary teeth; multiple rows of teeth on upper jaw; four compound teeth on vomer; teeth on lower jaw multiserial, those on inner row slightly enlarged, and 122-124 total vertebrae. Components of the variation of Dysomma taiwanense originally described from Taiwan are provided based on an additional 52 recently identified specimens.


Alpha-1-antitrypsin in liver disease.
Low alpha-1-antitrypsin (AAT) levels are known to be associated with liver disease. As AAT is also synthesised in the liver, we investigated whether liver disease itself may result in low AAT levels. AAT was measured in plasma from 100 patients with various liver diseases including hepatitis, cirrhosis, jaundice and liver failure. Twenty-eight patients had increased AAT values (greater than 3.1 g/L), 70 had normal AAT values (between 1.5 and 3.1 g/L) and 2 had decreased AAT levels (less than 1.5 g/L). The 2 patients with low AAT levels were found to be of the PiMZ phenotype. There was no significant correlation between any of the standard 'liver function tests' and the AAT level. Our findings suggest that in liver disease AAT levels are usually normal or increased. Low levels are uncommon and the possibility of an abnormal AAT phenotype being associated with the liver disease should be examined.


Benzylpiperidine-Linked Diarylthiazoles as Potential Anti-Alzheimer's Agents: Synthesis and Biological Evaluation.
A novel series of hybrid molecules were designed and synthesized by fusing the pharmacophoric features of cholinesterase inhibitor donepezil and diarylthiazole as potential multitarget-directed ligands for the treatment of Alzheimer's disease (AD). The compounds showed significant in vitro anticholinesterase (anti-ChE) activity, the most potent compound (44) among them showing the highest activity (IC50 value of 0.30 ± 0.01 μM) for AChE and (1.84 ± 0.03 μM) for BuChE. Compound 44 showed mixed inhibition of AChE in the enzyme kinetic studies. Some compounds exhibited moderate to high inhibition of AChE-induced Aβ1-42 aggregation and noticeable in vitro antioxidant and antiapoptotic properties. Compound 44 showed significant in vivo anti-ChE and antioxidant activities. Furthermore, compound 44 demonstrated in vivo neuroprotection by decreasing Aβ1-42-induced toxicity by attenuating abnormal levels of Aβ1-42, p-Tau, cleaved caspase-3, and cleaved PARP proteins. Compound 44 exhibited good oral absorption and was well tolerated up to 2000 mg/kg, po, dose without showing toxic effects.


Aging and vitamin E deficiency are responsible for altered RNA pathways.
Fibrillar centers (FCs), dense fibrillar (DFC) and granular (GC) components in nucleoli, and perichromatin granules (PGs) in nucleoplasm were measured by morphometry. FC size and their nucleolar surface fraction significantly decreased in aging and vitamin E deficiency. The GC and DFC nucleolar fraction was unchanged in adult and old rats, but in vitamin E-deficient animals GC increased and DFC decreased significantly. PG density significantly increased in aging and decreased in vitamin E deficiency. The quantitative evaluation of immunolabeled transcription and splicing factors revealed that polymerase II and SC-35 significantly decreased in old and vitamin E-deficient versus adult animals. Fibrillarin and snRNPs did not change between adult and old rats, but were significantly lower in vitamin E-deficient rats. These data document altered RNA pathways in aging and vitamin E deficiency. Considering the antioxidant role of vitamin E, they lend further support to the importance of free radical production and control in the aging process.


PVR/CD155 Ala67Thr Mutation and Cleft Lip/Palate.
The 19q13 locus has been linked to cleft lip and palate by our group and independently by others. Here we fine mapped the region in an attempt to identify an etiological variant that can explain cleft lip and palate occurrence. A total of 2739 individuals born with cleft lip and palate, related to individuals born with cleft lip and palate, and unrelated were studied. We used linkage and association approaches to fine map the interval between D19S714 and D19S433 and genotypes were defined by the use of TaqMan chemistry. We confirmed our previous findings that markers in PVR/CD155 are associated with cleft lip and palate. We studied the mutation Ala67Thr further and calculated its penetrance. We also attempted to detect PVR/CD155 expression in human whole saliva. Our results showed that markers in PVR/CD155 are associated with cleft lip and palate and the penetrance of the Ala67Thr is very low (between 1% and 5%). We could not detect PVR/CD155 expression in adult human whole saliva and PVR/CD155 possibly interacts with maternal infection to predispose children to cleft lip only.


Room-temperature decay and light reactivation of high-Tc ferromagnetism in an oxide-diluted magnetic semiconductor.
We present a novel route for manipulation of the ferromagnetic order in Co-doped TiO2 using UV laser irradiation. The ferromagnetic order of the nanocrystal films decays with aging in air at room temperature, which can be reactivated and enhanced by UV irradiation, whereas the coercive force reduces with irradiation time. Photoinduced trapped electrons were suggested to induce the ferromagnetic order. We believe that light manipulation is a general method for tuning the magnetic properties of oxide-based diluted magnetic semiconductors, which can find practical applications in future integrated magneto-optical nanoelectronics.


Cation exchange in mammalian erythrocytes. III. The prolytic effect of x-rays on human cells.
Freshly drawn heparinized human whole blood is exposed to x-rays in amounts up to 54,000 r in vitro and then equilibrated under a controlled atmosphere at 24 or 38 degrees C. For as long as 26 hours following exposure, potassium is progressively lost from the cells and quantitatively replaced by sodium with little, if any, osmotic disturbance. The mean rate of loss at 20,000 r and 24 degrees C. is about 0.4 per cent of the initial cell potassium per hour and approximately doubles for a 20,000 r increase. It is accentuated if blood is stored at low temperature (5 degrees C.) following radiation exposure. Isotope experiments show that the rate of entrance of potassium into the cells is practically unaltered, the principal effect being an acceleration of the rate from cells to plasma. This suggests that radiation may have interfered with a mechanism of selective potassium accumulation based on preferential retention of the element. The sodium which enters the cells following irradiation contributes to the rapidly exchanging portion of the cellular sodium, suggesting that this fraction is ionic sodium.


Differential expression of antimicrobial polypeptides in cord blood samples of preterm and term infants.
To determine levels of antimicrobial polypeptides (AMP) in cord blood of term and preterm neonates and to investigate influencing factors. In a single-centre study, n = 139 preterm infants and n = 36 term infants were included. AMP levels were analysed in supernatants of whole cord blood cultures with a standardised concentration of 5 × 10(6) white blood cells/mL via enzyme-linked immunosorbent assay (ELISA). Lactoferrin, human neutrophil peptides (HNP) 1-3 and bacterial permeability-increasing protein (BPI) expression in cord blood of preterm infants were influenced by the cause of preterm delivery, that is increased levels in infants with clinical amniotic infection. AMP levels also weakly correlated with white blood cell and neutrophil count at birth. In the whole cohort, no association between gestational age or birthweight with AMP levels was found. In the subgroup of infants without clinical amniotic infection (n = 77 preterm infants, n = 36 healthy term infants), we noted a weak correlation between gestational age and lactoferrin, calprotectin and HNP1-3 levels. In addition to that, we observed higher levels of lactoferrin and HNP1-3 in large-for-gestational-age infants. Our study confirms that several factors influence cord blood AMP levels which underlines the difficulties of using AMP levels as biomarkers of immunological response.


Expression and function of miR-155 in rat synovial fibroblast model of rheumatoid arthritis.
Rheumatoid arthritis (RA) is a common autoimmune disease characterized by joint synovial inflammation and is a challenge for researchers and clinicians. MicroRNAs (miRNAs/miRs) represent a group of small non-coding RNA molecules that post-transcriptionally regulate mRNA expression and are involved in various diseases, including cancer, autoimmune and metabolic diseases, as well as neurological disorders. In the present study, various experiments were performed to investigate the effects and underlying mechanism of miR-155 in RA using rat synoviocytes induced by lipopolysaccharide (LPS) to model rheumatoid arthritis. It was revealed that synovial fibroblasts exhibited significantly higher miR-155 mRNA levels than the control group. Compared with the RA group, the viability of synovial fibroblasts was significantly decreased in the miR-155 mimics + RA group, but markedly increased in the miR-155 inhibitor + RA group. Compared with that in the RA + NC mimic or RA + NC inhibitor groups, the apoptosis of synovial fibroblasts increased significantly in the miR-155 mimics + RA group, but was significantly decreased in the miR-155 inhibitor + RA group. The miR-155 mimics + RA group exhibited higher expression levels of β-catenin, matrix metalloproteinase 7 and cyclin D1 compared with the miR-155 inhibitor + RA group, and the glycogen synthase kinase protein levels was lower compared with the miR-155 inhibitor + RA group. In brief, it was inferred that the Wnt signaling pathway is involved in the miR-155-associated inhibition of RA synovial fibroblast viability and induction of cell apoptosis. Inhibition of miR-155 may be an effective treatment for RA through regulation of the Wnt signaling pathway to reduce cell apoptosis and enhance cell viability.


Influence of alpha-tocopherol on prostacyclin synthesis by rat's arterial and myometrial tissues.
Influence of alpha-tocopherol on PGI2 synthesis by rat arterial and myometrial tissues was investigated using a rat platelet antiaggregatory bioassay. Chronic administration of alpha-tocopherol to female rats (10 mg kg-1 day-1 s.c. for 14 days) significantly increased ex-vivo PGI2 synthesis by the arterial tissue from 12.7 +/- 0.3 (control, mean +/- s.e.m) to 17.2 +/- 0.4 ng PGI2 mg-1 wet tissue and by the myometrial tissue (in proestrus) from 1.1 +/- 0.07 (control) to 1.85 +/- 0.1 ng PGI2 mg-1 wet tissue (P less than 0.05, n = 6). alpha-tocopherol (5 mg kg-1 day-1 for 14 days) did not stimulate PGI2 to any significant level. Pretreatment of male rat arterial tissue with alpha-tocopherol (0.02, 0.1 or 0.2 mM) in vitro increased PGI2 synthesis in a dose-dependent manner. At a dose of 0.2 mM it increased PGI2 synthesis from 13.70 +/- 0.70 (control) to 22.6 +/- 1.4 ng PGI2 mg-1 wet tissue (P less than 0.1, n = 6). Pre-treatment of 14-day pregnant rat myometrium with alpha-tocopherol 0.2 and 0.4 mM significantly increased PGI2 synthesis from 1.2 +/- 0.06 (control) to 1.90 +/- 0.12 and 2.1 +/- 0.1 ng PGI2 mg-1 wet tissue, respectively (P less than 0.05, n = 6). The results indicate that the ability of alpha-tocopherol to stimulate PGI2 synthesis may partly contribute towards better understanding of the mechanisms that underly the protective effect of alpha-tocopherol against experimentally induced decreases in coronary flow and intravascular coagulations in some mammals. Furthermore adequate intake of alpha-tocopherol during pregnancy may enhance uterine blood flow and ensure adequate foetal nutrition.


Otolaryngological manifestations and genetic characteristics of lipoid proteinosis.
We investigated the otolaryngological manifestations and genetic characteristics of lipoid proteinosis (LP). Twenty-two cases of LP were included. Multisystem manifestations, pharyngolaryngeal behaviors, and histopathologic and genetic characteristics were evaluated. All patients presented with hoarseness as the first sign. The onset was in the newborn period in 14 cases. Video-laryngoscopy revealed irregularities along the edges and the surface of the vocal folds with diffuse yellowish deposits. In 18 cases, the oral and pharyngeal mucosa showed yellow-white infiltrates. In all cases, whitish moniliform papules were observed on the upper eyelids. In 18 cases, pocklike scars were present on the face and limbs. Histologically, there were diffuse depositions of hyaline material in the submucosa and dermis that were positive on periodic acid-Schiff staining and negative on Congo red staining. Two patients had homozygous mutations in ECM1 exon 6 and exon 8, and 13 patients had heterozygous mutations in ECM1 exons. Nine patients underwent microlaryngosurgery for excision of deposits in the vocal folds. The voice function was significantly improved after operation. Persistent hoarseness from early infancy with laryngeal involvement is the most common feature of LP. Histopathologic characteristics and skin and mucosa changes aid in the diagnosis. Microlaryngosurgery may improve the voice function. Compound heterozygote mutations may be frequent in LP in nonconsanguineous Chinese families.


[Epidemiologic aspect of lyme borreliosis among the inhabitants of Podlasie Province].
The aim of this study was to evaluate Lyme borreliosis morbidity among the inhabitants of Podlasie Province in the years of 1996-2000. The new cases of Lyme borreliosis reported in the years of 1996-2000 in Podlasie Province were analyzed. The analysis included the data from Mz-56 and Mz-57 forms and the information from epidemiologic investigations sent to Regional Sanitary and Epidemiologic Station in Bialystok. In 1996-2000, screening examinations were carried out in the same group of 358 forestry workers including 44 (12.29%) females and 314 (87.71%) males aged from 21 to 64 (x = 41.2) years. Immunoenzymatic test of ELISA Borrelia recombinant IgM and IgG by Biomedica firm (Austria) were used to detect B. burgdorferi antibodies. In 1996-2000, 4933 of borreliosis cases were registered in Poland including 1377 (27.91%) in Podlasie Province. The morbidity rate in Podlasie province ranged from 15.05% in 1996 year to 21.29% in 2000 year of the whole country morbidity. At the same time, the incidence rate in Podlasie Province ranged from 9.09 in 1996 to 32.2 in 2000 year and was 6.72-fold higher than the incidence rate in the whole country. In Eastern and Central region of the province, 80.54% of cases were registered whereas, 14.09% by the Lakeside of Augustów and Suwalki only 5.37% in the western region of the province. It was proved that the morbidity increased in proportion to the age of patients (41.39% of patients were at the age of 30-49) and it decreased only above the age of 60. Thus, Lyme borreliosis affects mainly people at working age. The increase in B. burgdorferi antibody detectability was noticed in the population of forestry workers; it was detected in 38.55% in 1995 and in 2000 in 43.56% of the examined. Interestingly, 81 people infected with B.b in 1995-2000 included 74 men and only 7 women, supporting the statement that B.b infection affects mainly working professionals and the risk increases with the practice. Our studies indicate that Lyme borreliosis is a serious health problem among the inhabitants of Podlasie Province which is an endemic area for Ixodes ricinus ticks infected with B. burgdorferi spirochete.


PEGylated Chromatography: Efficient Bioseparation on Silica Monoliths Grafted with Smart Biocompatible Polymers.
Novel oligo(ethylene glycol)-based thermoresponsive stationary phases have been studied for the separation of bioanalytes. Well-defined copolymers of (2-methoxyethoxy)ethyl methacrylate and oligo(ethylene glycol) methacrylate were synthesized by atom-transfer radical polymerization in the presence of an N-succinimidyl-functionalized initiator. The reactive chain ends of these copolymers were then reacted with amino-functionalized silica monoliths. The formed composites were studied as chromatography materials. For instance, it was demonstrated that thermoresponsive oligo(ethylene glycol)-based stationary phases allow rapid, efficient separation of steroid and protein mixtures in pure water under isocratic high-performance liquid chromatographic elution.


Quality improvement initiative for a sustained increase in human milk donation during the hospital stay.
The demand for donors' human milk is much more than the availability of the same due to the myriad challenges faced during the collection process. Baseline milk volume donation done in a human milk bank facility located in tertiary care government institute in a low-middle income country was assessed. It was initially aimed to increase the absolute quantity of milk volume donation by 30% over a period of 6 months, which was subsequently continued following COVID-19 emergence (over another 12 months) along with a particular emphasis on the sustenance of milk donation activities. Counselling of both the healthcare workers and stakeholders, standardising the timing of milk donation and other policies, equipment in proportion to demand and supply and addition of human resource were done as a multiprong approach to have sustained increase in human milk donation. The median control line (MCL) showed a shift of 27.8%; from a baseline of 5032 mL to 6971 mL during intervention phase I comprising of five plan do study act cycles spread over a period of 6 months. During the sustenance phase I even though the monthly collection was non-uniform, there was a further 16% upward shift in MCL to 8122 mL. During the second intervention phase, each component of the Ishikawa diagram was worked on (people, policy, place, procedure) resulting in a more than 100% increase from sustenance phase I taking the MCL to 17 181 mL with an overall increase of 3.41 times from the baseline. Dedicated counselling and constant motivation have been conventionally considered as the utmost measure to increase milk volume donation in milk bank. Our study highlights the need to introduce scheduled timings along with sufficient equipment and manpower to overcome the shortage of milk donation in human milk bank facilities.


Factors predictive of failure of Brescia-Cimino arteriovenous fistulas.
To evaluate patency rates of Brescia-Cimino fistulas and to find out which independent factors were predictors of failure. Retrospective clinical study. University hospital, The Netherlands. 150 consecutive patients (mean age 56 years, range 17-80) who had 153 primary Brescia-Cimino fistulas created during the 5-year period January 1995-December 1999. Patency rates calculated by the Kaplan-Meier method and the possible predictive value of 20 different variables assessed by Cox's proportional hazard model. The primary patency rate was 70% at 3 months, and 7 distinct factors were significantly associated with failure of the fistula. The ones with a hazard ratio (HR) for failure greater than 2.5 were: the start of dialysis before creation of the fistula (HR 2.79, p < 0.01), moderate or poor quality of both the artery (HR 2.54, p < 0.01) and vein (HR 3.55, p < 0.001), and postoperative use of acenocoumarol instead of acetylsalicylic acid (HR 3.14. p < 0.01). The major determinants for a successfully created Brescia-Cimino fistula were creation of the fistula before the start of dialysis, as well as good quality of both the artery and the vein. This argues for timely creation of such fistulas in patients with end-stage renal disease and for accurate preoperative examination to establish the quality of the vessels.


Nuclear localization signals: a driving force for nuclear transport of plasmid DNA in zebrafish.
Nuclear localization signals (NLSs) are short peptides required for nuclear transport of karyophilic proteins. We review in this paper how the nuclear targeting property of NLS peptides has been taken advantage of to enhance the efficiency of nuclear uptake of transgene DNA in zebrafish and how it may improve the efficiency of transgenesis in this species. Synthetic NLS peptides can bind to plasmid DNA by ionic interactions. Cytoplasmic injection of DNA-NLS complexes in zebrafish eggs enhances the rate and the amount of plasmid DNA taken up by embryonic nuclei. Nuclear import of DNA-NLS complexes has been duplicated in vitro and exhibits energetic and cytosolic requirements similar to those for nuclear protein import. Furthermore, binding NLSs to DNA increases expression frequency of the transgene. We suggest that NLS peptides may constitute a valuable tool to improve the efficiency of transgenesis in zebrafish and other species.


Engineering efficient termination of bacteriophage T7 RNA polymerase transcription.
The bacteriophage T7 expression system is one of the most prominent transcription systems used in biotechnology and molecular-level research. However, T7 RNA polymerase is prone to read-through transcription due to its high processivity. As a consequence, enforcing efficient transcriptional termination is difficult. The termination hairpin found natively in the T7 genome is adapted to be inefficient, exhibiting 62% termination efficiency in vivo and even lower efficiency in vitro. In this study, we engineered a series of sequences that outperform the efficiency of the native terminator hairpin. By embedding a previously discovered 8-nucleotide T7 polymerase pause sequence within a synthetic hairpin sequence, we observed in vivo termination efficiency of 91%; by joining 2 short sequences into a tandem 2-hairpin structure, termination efficiency was increased to 98% in vivo and 91% in vitro. This study also tests the ability of these engineered sequences to terminate transcription of the Escherichia coli RNA polymerase. Two out of 3 of the most successful T7 polymerase terminators also facilitated termination of the bacterial polymerase with around 99% efficiency.


[Effect of glutamic acid content of the diet on isotope-labeled glutamic acid catabolism in rats. 1. The course of 14CO excretion following intragastric administration of 14C-glutamic acid].
Male rats received in 8 groups of 10 animals each for a period of 7 days 7 synthetic diets and one semisynthetic diet on maintenance requirement level. A L-amino acid mixture corresponding to the pattern of egg protein without glutamic acid was the protein source of the synthetic diets. Glutamic acid was supplemented successively from 0 to 58 mol-% of the total amino acid content. The crude protein source of diet 8 was whole egg powder. On the 8th day of experiment 5 animals per group were labelled by intragastric infusion (i.g.) with 14C-U-glutamic acid. During the following 24 hours the excretion of CO2 and 14CO2 was measured. Throughout the experimental feeding body weight was relative constant, however, when the synthetic diets were fed it was necessary to increase the daily amount of energy from 460 to 480 kJ/kg0,67. The relative 14CO2-excretion within 24 hours was 68-75% of the dose. However, the main part of the amount of radioactivity excreted during 24 hours was found after 4 to 6 hours already. Exponential functions calculated from the data of cumulative 14CO2-excretion suggest the existence of a fast process of 14CO2-formation directly from 14C-glutamic acid, reaching a plateau within 2 hours and a slow process of oxidation of intermediates of glutamic acid metabolism, causing a continued 14CO2-formation even after 24 hours. The oxidation of 14C-glutamic acid to CO2 decreased 2 to 14 hours after i.g. labelling if the glutamic acid content of the diet increased. The same was found for the specific radioactivity of 14CO2. A storage of intermediates of glutamic acid before degradation was assumed.


Purification and characterization of recombinant Caulobacter crescentus Cu,Zn superoxide dismutase.
Recombinant Cu,Zn Superoxide Dismutase from Caulobacter crescentus has been expressed in Escherichia coli and characterized. The corresponding recombinant protein has a molecular weight typical of a homodimeric Cu,ZnSODs and an activity comparable to that of other prokaryotic enzymes. The copper active site is characterized by a peculiar axial geometry as evidenced by its electron paramagnetic resonance spectrum, moreover, the copper atom displays a low accessibility toward external chelating agents indicating a lower solvent accessibility when compared to other prokaryotic enzymes. Investigation of the enzyme thermal stability through differential scanning calorimetry indicates the occurrence of two transitions at low and higher temperature that are found to be due to the apo and holo protein, respectively, confirming that the metals have a crucial role in the stabilization of this class of enzymes.


Contemporary issues in the management of childhood brain tumors.
The incidence of childhood brain tumors appears to be on the rise caused in part by improvements in neuroimaging and its increased availability. Current protocols for treatment of childhood brain tumors are utilizing chemotherapy more extensively in addition to and in lieu of radiotherapy to improve survival for tumors such as medulloblastoma and high grade astrocytoma, and to lessen treatment related morbidity in infants with malignant brain tumors and in older children with standard risk medulloblastomas and germ cell tumors. The increasing understanding of the molecular events that lead to oncogenesis will help unravel the causes and improve therapy for a group of illnesses that cause significant neurologic morbidity and mortality.


Characterization and evaluation of olive germplasm in southern Italy.
The southern Italian region of Campania has suitable pedo-climatic conditions and a large varietal heritage able to produce oils with high typicality. The aim of this study was to characterize 20 cultivars belonging to Campania's olive germplasm, evaluating their vegetative and production aspects and their oil quality characteristics. The study was conducted from 2003 to 2009, observing the following aspects in six plants per variety: entry into production, vigour, ripening and drupe oil content. The following analyses were carried out on monovarietal oils, obtained by microextractor: acid composition, polyphenol content and aromatic profile. The agronomic results showed early entry into production for the cultivars Racioppella, Ortolana, Biancolilla and Carpellese. However, entry into production was delayed for Ritonnella, Ortice, Cornia and Rotondella. As regards vegetative behaviour, Asprinia, from the province of Caserta and Pisciottana and Carpellese, from the province of Salerno, proved the most vigorous cultivars. Groups of similar cultivars emerged from chemical and sensory analysis of their oils, while other accessions were well characterized and separated from each other, showing a high level of diversity and specificity.


Pediatric septic shock: recognition and management in the emergency department.
Sepsis is a leading cause of morbidity and mortality in children. Early recognition and timely initiation of empiric broadspectrum antibiotics and crystalloid fluid administration have been associated with better outcomes. Although evidence for diagnosis and treatment of septic shock was first generated in adult studies, it is clear that pediatric studies are needed for management of septic shock in children. This issue provides guidance for managing septic shock in children, with a focus on early recognition and appropriate resuscitation.


Respiratory viral infections during the 2009-2010 winter season in Central England, UK: incidence and patterns of multiple virus co-infections.
Acute viral respiratory infections are the most common infections in humans. Co-infection with different respiratory viruses is well documented but not necessarily well understood. The aim of this study was to utilise laboratory data from the winter season following the 2009 influenza A(H1N1) outbreak to investigate rates of respiratory virus co-infections, virus prevalence in different age groups and temporal variations in virus detection. The Health Protection Agency Public Health Laboratory (HPA PHL) Birmingham, UK, routinely uses polymerase chain reaction (PCR) to detect common respiratory viruses. The results from specimens received for respiratory virus investigations from late September 2009 to April 2010 were analysed. A total of 4,821 specimen results were analysed. Of these, 323 (13.2 %) had co-detections of two viruses, 22 (0.9 %) had three viruses and four (0.2 %) had four viruses. Reciprocal patterns of positive or negative associations between different virus pairs were found. Statistical analysis confirmed the significance of negative associations between influenza A and human metapneumovirus (HMPV), and influenza A and rhinovirus. Positive associations between parainfluenza with rhinovirus, rhinovirus with respiratory syncytial virus (RSV) and adenovirus with rhinovirus, parainfluenza and RSV were also significant. Age and temporal distributions of the different viruses were typical. This study found that the co-detection of different respiratory viruses is not random and most associations are reciprocal, either positively or negatively. The pandemic strain of influenza A(H1N1) was notable in that it was the least likely to be co-detected with another respiratory virus.