Δευτέρα, 15 Μαΐου 2017

Keep within target range of blood oxygen levels, new guideline recommends

Health professionals should always specify a target saturation range when prescribing oxygen and should then ensure that patients are monitored to avoid high or low levels that can cause serious...
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Fixed second appointment improves breast screening attendance

Women who miss a breast screening appointment should be given a second appointment with a fixed date and time, researchers recommend in Lancet Oncology.1 Their study found that this approach was...
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Missense variant pathogenicity predictors generalize well across a range of function-specific prediction challenges

ABSTRACT

The steady advances in machine learning and accumulation of biomedical data have contributed to the development of numerous computational models that assess the impact of missense variants. Different methods, however, operationalize impact differently. Two common tasks in this context are the prediction of the pathogenicity of variants and the prediction of their effects on a protein's function. These are related but distinct problems and it is unclear whether methods developed for one are optimized for the other. The Critical Assessment of Genome Interpretation (CAGI) experiment provides a means to address this question empirically. To this end, we participated in various protein-specific challenges in CAGI with two objectives in mind. First, to compare the performance of methods in the MutPred family with the state-of-the-art. Second and more importantly, to investigate the applicability of general-purpose pathogenicity predictors to the classification of specific function-altering variants without additional training or calibration. We find that our pathogenicity predictors performed competitively with other methods, outputting score distributions in agreement with experimental outcomes. Overall, we conclude that binary classifiers learned from disease-causing mutations are capable of modeling important aspects of the underlying biology and the alteration of protein function resulting from mutations.

This article is protected by copyright. All rights reserved



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Laparoscopic Transhiatal Esophagectomy

Abstract

The operation has three parts:



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Malignancy Incidence, Management, and Prevention in Patients with Rheumatoid Arthritis

Abstract

Traditional and biologic disease-modifying antirheumatic drugs (DMARDs) are effective medications for the management of rheumatoid arthritis (RA). However, the effects of these medications on immune function raises concern that they may increase long-term cancer risk. The baseline risk for some cancers appears to differ in patients with RA compared to the general population, with the former having an increased risk of lymphoma, lung cancer and renal cancer, but a decreased risk of colorectal and breast cancer. Some DMARDs appear to increase the rate of specific cancer types (such as bladder cancer with cyclophosphamide), but few appear to increase the overall cancer risk. Studying the link between lymphoma and disease severity in RA is complicated because patients with persistently active disease are at increased risk for lymphoma, and disease severity correlates with more intense use of immunosuppressive medications. Overall, cancer risk in patients with RA is slightly above that of the general population, with the increased risk likely secondary to an increased risk of lymphomas in those with high disease activity. Risk mitigation includes management of RA disease activity as well as age- and sex-appropriate cancer screening.



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A rounded opacity silhouetting the left heart border and hilum

A 73-year-old woman with hypertension and atrial fibrillation presented with head and neck injury after mechanical fall. During workup, chest X-ray anteroposterior view (figure 1) revealed a rounded opacity silhouetting the left heart border and hilum. Subsequent contrast-enhanced CT of the chest showed single, 6.4 cm, rounded, well-defined, thin-walled, non-enhanced, low attenuated (–20 and 20 Hounsfield Unit) and homogenous cyst-like structure at the left mediastinum connected to pericardial recesses and not attached to adjacent structures (figure 2A–C). Transthoracic echocardiogram ruled out left ventricular aneurysm, aortic aneurysm, solid tumour and outflow tracts obstruction. Although bronchogenic cyst, oesophageal duplication cyst, thymic tumour and mediastinal lymphoma were considered as possible differentials, radiological features such as CT appearance, homogenous attenuation, unrelated to the underlying structures favoured pericardial cyst. Since patient was asymptomatic, patient and family member were unwilling to undergo surgical removal and pathological confirmation. Follow-up with non-enhanced CT of...

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182
6948891480

Distinct phosphodiesterase 5A-containing compartments allow selective regulation of cGMP-dependent signalling in human arterial smooth muscle cells

Publication date: August 2017
Source:Cellular Signalling, Volume 36
Author(s): Lindsay S. Wilson, Manhong Guo, M. Bibiana Umana, Donald H. Maurice
Cyclic GMP (cGMP) translates and integrates much of the information encoded by nitric oxide (NO·) and several natriuretic peptides, including the atrial natriuretic peptide (ANP). Previously, we reported that integration of a cGMP-specific cyclic nucleotide phosphodiesterase, namely phosphodiesterase 5A (PDE5A), into a protein kinase G (PKG)- and inositol-1,4,5-trisphosphate receptor (IP3R)-containing endoplasmic reticulum (ER) signalosome allows localized control of PDE5A activity and of PKG-dependent inhibition of IP3-mediated release of ER Ca2+ in human platelets. Herein, we report that PDE5A integrates into an analogous signalosome in human arterial smooth muscle cells (HASMC), wherein it regulates muscarinic agonist-dependent Ca2+ release and is activated selectively by PKG-dependent phosphorylation. In addition, we report that PDE5A also regulates HASMC functions via events independent of PKG, but rather through actions coordinated by competitive cGMP-mediated inhibition of cAMP hydrolysis by the so-called cGMP-inhibited cAMP PDE, namely phosphodiesterase 3A (PDE3A). Indeed, we show that ANP increases both cGMP and cAMP levels in HASMC and promotes phosphorylation of vasodilator-stimulated phospho-protein (VASP) at each the PKG and PKA phospho-acceptor sites. Since selective inhibition of PDE5 decreased DNA synthesis and chemotaxis of HASMC, and that PDE3A knockdown obviated these effects, our findings are consistent with a role for a PDE5A-PDE3A-PKA axis in their regulation. Our findings provide insight into the existence of distinct “pools” of PDE5A in HASMC and support the idea that these discrete compartments regulate distinct cGMP-dependent events. As a corollary, we suggest that it may be possible to target these distinct PDE5A-regulated pools and in so-doing differentially impact selected cGMP-regulated functions in these cells.



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Direct and indirect activation of eukaryotic elongation factor 2 kinase by AMP-activated protein kinase

Publication date: August 2017
Source:Cellular Signalling, Volume 36
Author(s): M. Johanns, S. Pyr dit Ruys, A. Houddane, D. Vertommen, G. Herinckx, L. Hue, C.G. Proud, M.H. Rider
BackgroundEukaryotic elongation factor 2 (eEF2) kinase (eEF2K) is a key regulator of protein synthesis in mammalian cells. It phosphorylates and inhibits eEF2, the translation factor necessary for peptide translocation during the elongation phase of protein synthesis. When cellular energy demand outweighs energy supply, AMP-activated protein kinase (AMPK) and eEF2K become activated, leading to eEF2 phosphorylation, which reduces the rate of protein synthesis, a process that consumes a large proportion of cellular energy under optimal conditions.AimThe goal of the present study was to elucidate the mechanisms by which AMPK activation leads to increased eEF2 phosphorylation to decrease protein synthesis.MethodsUsing genetically modified mouse embryo fibroblasts (MEFs), effects of treatments with commonly used AMPK activators to increase eEF2 phosphorylation were compared with that of the novel compound 991. Bacterially expressed recombinant eEF2K was phosphorylated in vitro by recombinant activated AMPK for phosphorylation site-identification by mass spectrometry followed by site-directed mutagenesis of the identified sites to alanine residues to study effects on the kinetic properties of eEF2K. Wild-type eEF2K and a Ser491/Ser492 mutant were retrovirally re-introduced in eEF2K-deficient MEFs and effects of 991 treatment on eEF2 phosphorylation and protein synthesis rates were studied in these cells.Results & conclusionsAMPK activation leads to increased eEF2 phosphorylation in MEFs mainly by direct activation of eEF2K and partly by inhibition of mammalian target of rapamycin complex 1 (mTORC1) signaling. Treatment of MEFs with AMPK activators can also lead to eEF2K activation independently of AMPK probably via a rise in intracellular Ca2+. AMPK activates eEF2K by multi-site phosphorylation and the newly identified Ser491/Ser492 is important for activation, leading to mTOR-independent inhibition of protein synthesis. Our study provides new insights into the control of eEF2K by AMPK, with implications for linking metabolic stress to decreased protein synthesis to conserve energy reserves, a pathway that is of major importance in cancer cell survival.

Graphical abstract

image


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Transient hemiparaesthesias and dysarthria

A previously healthy 29-year-old Mexican woman presented to an emergency department with transient hemiparaesthesias and dysarthria. There was no evidence of stroke on cross-sectional imaging of the head, and she was discharged without a clear diagnosis. Two days later, she returned with acute abdominal pain. Abdominal imaging revealed complete occlusion of the right renal artery, prompting emergency embolectomy. Following the procedure, she developed acute haemoptysis, dyspnoea and hypoxaemia. Chest imaging demonstrated evidence of pulmonary venous hypertension. Cardiac auscultation revealed an opening snap followed by a diastolic murmur with presystolic accentuation. These sounds were better appreciated in combination with phonocardiography, a technique supplanted by echocardiography in the 1970s1 that visualised heart sounds (video 1). An echocardiogram confirmed the presence of mitral stenosis (MS), unifying the syndrome of embolic phenomena, haemoptysis and pulmonary hypertension. She underwent successful mitral valve replacement and has since returned to normal...


Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182
6948891480

Signal and regulatory effects of methylglyoxal in eukaryotic cells (review)

Abstract

Methylglyoxal (MG) is one of the physiological glucose metabolites formed in living organisms. The data on the influence of MG on different internal systems of eukaryotic cells, including the central signaling pathways, are been discussed in the review. The central signaling pathways are stress-activated and sensitive to the action of reactive oxygen species. Integration of the literary data and authors' results has allowed the conclusion that MG action on cells is multidirectional and is determined by its concentration and the physiological state of the cell. The cellular reaction upon increasing MG concentrations has a phase pattern and can be described by the hormesis concept. It has been hypothesized that MG participates in the formation of the braking regulatory circuit, which modulates the sensitivity of hypothalamus neurons to glucose. It is concluded that MG has a possible role in the functioning of the great biological clock. We propose that the data discussed in this review allow methylglyoxal to be considered a molecule with signal and regulatory functions.



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Penicillium verrucosum as promising candidate for bioremediation of environment contaminated with synthetic detergent at high concentration

Abstract

The aim of this study was to investigate the effect of synthetic detergent Merix (Henkel, Kruševac, Serbia), and its particular components—ethoxylated oleyl-cetyl alcohol and sodium tripolyphosphate on the growth and metabolic activity of Penicillium verrucosum. During 19 days of fungal cultivation in Czapek-Dox liquid medium supplemented with or without 0.5% pollutants, the following parameters were observed: pH, the total biomass dry weight, the quantity of free and total organic acids and proteolytic activity. The detergent caused a slight stimulatory (2.41%) effect whereas sodium tripolyphosphate and ethoxylated oleyl-cetyl alcohol provided a slight inhibitory action (0.59 and 2.75%, respectively) on the fungal biomass. The pollutants decreased pH values of the media and the quantity of free organic acids. In contrast, they enhanced the quantity of total organic acids. Proteolytic activity remained nearly unchanged (95.8%) in the presence of detergent and reduced to 80.1% in sodium tripolyphosphate-supplemented medium. In contrast, the enzyme activity sharply increased (260.8%) with ethoxylated oleyl-cetyl alcohol. The obtained results indicate the potential of P. verrucosum in bioremediation of environment contaminated with synthetic detergent taken in high concentration.



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Synonymous codon changes at the 5′-end of the gene strongly impact the heterologous protein expression in Escherichia coli

Abstract

This study describes the impact of 5′-end codon modulation on the expression of a heterologous gene, human granulocyte colony stimulating factor (GCSF), in Escherichia coli. Fourteen different constructs (pGCSF-01 to pGCSF-14) carrying single or multiple synonymous substitutions at +2, +3 and further down from +4 to +7 codons, were prepared and their expression was monitored in E. coli BL21 Codon-Plus (DE3) RIPL using a strong T7 lac-promoter based expression system. A single nucleotide change at +2 Thr codon (ACC→ACA) either alone or in combination with +3 Pro codon (CCC/CCT/CCA) resulted in the expression enhancement of an otherwise poorly expressed native-GCSF, to a level that corresponded to 45–50% of the total E. coli BL21 CodonPlus (DE3) RIPL cellular proteins. The differences in GCSF expression amongst different constructs could be attributed to the preferential or non-preferential codon usage, reduced number of G/C nucleotides and the stability of mRNA secondary structure formed near the 5′-end coding region. The expression of GCSF achieved was in the form of biologically inactive inclusion bodies that were solubilized using mild concentration of a non-ionic surfactant and refolded by a simplified, step-dialysis approach. Biological activity of the purified GCSF, assessed in induced neutropenic mice, was similar to the commercially available preparation of the GCSF analog (filgrastim).



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Flor yeast strains from culture collection: Genetic diversity and physiological and biochemical properties

Abstract

Sixteen flor yeast strains from the Magarach Collection of the Microorganisms for Winemaking (Yalta, Crimea), which are used for production of sherry, were analyzed for morphophysiological, cultural, and biochemical properties. Long-term storage did not affect their viability or the preservation of major properties, such as their flor- and aldehyde-forming abilities, and the ability to produce wines with typical sherry properties. Significant variation in the strains was observed mainly in the aldehyde-forming and flor-forming abilities and flor properties. Interdelta typing was shown to be the most informative technique to study the genetic diversity of flor yeast strains. Certain correlations between genetic polymorphisms and the enological properties of the strains were observed. The presence of a 24-bp long deletion in the ITS1 spacer of the ribosomal gene cluster, a typical feature of Spanish flor yeast strains, is correlated with a high level of production of aldehydes and acetales, efficient flor formation, and the ability to produce high quality sherry. The presence of a specific deletion in the promoter of the FLO11 gene appeared to be less informative, since the aldehyde and acetal production and flor formation abilities of such strains were variable. The studies of intraspecies genetic polymorphism by various molecular markers have revealed a high degree of phylogenetic closeness of some yeast flor strains from different geographic regions.



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Effect of extra- and intracellular sources of CO 2 on anaerobic utilization of glucose by Escherichia coli strains deficient in carboxylation-independent fermentation pathways

Abstract

The effect of extra- and intracellular CO2 sources on anaerobic glucose utilization by Escherichia coli strains deficient in the main pathways of mixed acid fermentation and possessing a modified system of glucose transport and phosphorylation was studied. Intracellular CO2 generation in the strains was ensured resulting from the oxidative decarboxylation of pyruvic acid by pyruvate dehydrogenase. Endogenous CO2 formation by pyruvate dehydrogenase stimulated anaerobic glucose consumption by the strains due to the involvement in the fermentation process of condensation reactions between oxaloacetic acid and acetyl-CoA. The availability of an external CO2 source (dissolved in medium sodium bicarbonate) promoted utilization of carbohydrate substrate by favoring the predominant participation in the fermentation of reactions directly dependent on phosphoenolpyruvate carboxylation. The positive effect of the availability of exogenous СО2 was sharply decreased in recombinant strains with the impaired functionality of the reductive branch of the tricarboxylic acid cycle. As a result, intracellular СО2 generation coupled to acetyl-CoA formation promoted anaerobic glucose utilization by cells of the corresponding mutants more markedly than the presence in the medium of dissolved sodium bicarbonate.



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Protective effect of inhibitors of succinate dehydrogenase on wheat seedlings during osmotic stress

Abstract

The effect of malonate and sedaxane, a compound with the fungicidal effect which act as succinate dehydrogenase inhibitors, on the resistance of etiolated wheat seedlings (Triticum aestivum L.) to osmotic stress caused by 12% PEG 6000 solution, was studied. The presowing treatment of seeds with 0.3 mM sedaxane solution significantly reduced the inhibitory effect of osmotic stress on seedling growth. The protective effect of 10 mM malonate was significant when it was added to the incubation medium of the roots; the effect of preseeding treatment with malonate was less significant. Unlike malonate, malate had no positive effect on seedling growth under osmotic stress. The activity of succinate dehydrogenase and the hydrogen peroxide content decreased in seedlings after the treatment of roots with malonate and sedaxane. Pretreatment with sedaxane and the addition of malonate to the incubation medium of roots prevented the accumulation of a lipid peroxidation product, malondialdehyde, which is caused by osmotic stress, and increased peroxidase activity. It was concluded that the stress-protective effect of sedaxane and malonate on wheat seedlings might be due to the inhibition of succinate dehydrogenase-dependent formation of reactive oxygen species and the prevention of oxidative cell damage.



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Preparation, properties and potential applications of exopolysaccharides from bacteria of the genera Xanthobacter and Ancylobater

Abstract

Two new bacterial biopolymers (exopolysaccharides), ancylan and xylophilan, have been isolated and characterized. The optimal parameters for ancylan and xylophilan production under laboratory conditions were selected. Their physicochemical properties and effects on microorganisms (bacteria, fungi, and ciliates) were studied. The results suggest the potential application of these new exopolysaccharides in medicine and veterinary science.



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New recombinant producer of human ω-amidase based on Escherichia coli

Abstract

A new artificial gene encoding human ω-amidase (Nit2) adapted for highly efficient expression in E. coli has been established. A pQE-Nit2 plasmid construct controlled by the T5 promoter has been engineered for its expression. The nit2 gene within the pQE-Nit2 construct has optimized codon usage and an artificial 6His-tag sequence inserted directly after the ATG initiation codon. This tag provides the possibility of single-step purification of a product via metal chelate chromatography. The codon-usage optimization involves the inclusion of several codons of extremely rare occurrence in natural E. coli ORFs within a 30 a.a-long N-terminal region. Other codons included in the N-terminus have moderate occurrence in E. coli. The subsequent sequence of the artificial gene has been composed of the most frequently occurring codons in E. coli. The recombinant producer based on the pQE-Nit2 construct allowed purification of the enzyme with an activity of 6.2 ± 0.2 μmol/min/mg protein, which corresponds to or slightly exceeds the specific activity of rat liver Nit2. The omega-amidase preparation is necessary for the screening of potential inhibitors that can be used as candidate drugs to cure hyperammonemia disorders in liver pathologies and oncological diseases.



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Cyanide detoxification by microbial consortia of natural-industrial complexes of gold heap leaching

Abstract

Microorganisms that have adapted not only to high concentrations of pollutants but also to environmental conditions develop in autochthonous microbial communities of natural-industrial complexes of gold heap leaching. The biotechnological potential and diversity of autochthonous microbial communities involved in cyanide detoxification was evaluated by the example of a deposit situated in the Sakha (Yakutia) Republic. Under the zoning conditions of the ore heap, the biological component had a greater impact on cyanide destruction than chemical transformation. Metabolically active representatives of a microbial consortium are capable of surviving developed under these conditions. Phylotypes of the genus Serratia and family Alcaligenaceae that are capable of cyanide destruction and are potentially promising for the detoxification of wastes of gold heap leaching were revealed.



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Intermicrobial relationships of the pea nodule symbiont Serratia sp. Ent16 and its colonization of the host endorhizosphere

Abstract

A strain of Serratia sp. Ent16 isolated from internal tissues of pea nodule inhibited in vitro growth of the plant pathogens Fusarium oxysporum and Bipolaris sorokiniana and the model strain Rhizobium leguminosarum bv viceae 1078 but had a considerably weaker antagonistic effect on the Rhizobium strain Rh16 from its own nodule. Cells of the Ent16 strain tagged by the gfp gene (the Ent16-gfp strain) were not seen in the pea endorhizosphere when plants were grown in a rich culture medium. The development of symbiosis was favored by plant germination on filter paper. Confocal microscopy showed that individual cells of the Ent16-gfp strain were attached to the outer side of root hair cell walls, while agglomerations of fluorescent bacterial cells were detected in the zone of exoderm of lateral root formation and in root vessels. A series of scanned sections of pea root revealed the presence of the Ent16-gfp strain in lateral root primordia, through which the bacteria penetrated the endorhizosphere.



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Identification of the di- n -butyl phthalate-biodegrading strains and the biodegradation pathway in strain LMB-1

Abstract

DNA isolated from a greenhouse soil (Nanjing, Jiangsu Province, China) was suitable for PCR amplification of gene segment coding for the 16S rRNA. Diverse PCR products were characterized by cloning and sequencing, and analysis of bacterial colonies showed the presence over 26 phyla. The most bacteria belonged to Proteobacteria, Actinobacteria, Gemmatimonadetes, Acidobacteria and Planctomycetes. Furthermore, after the enrichment procedure of DBP-degrading microorganisms, 4 strains were isolated from the soil sample with di-n-butyl phthalate (DBP) biodegradability, and they were identified to be Rhizobium sp., Streptomyces sp., Pseudomonas sp. and Acinetobacter sp. Analysis of the degradation products by LC-MS led to identification of metabolites of DBP in strain LMB-1 (identified as Rhizobium sp.) which suggests that DBP was degraded through β-oxidation, demethylation, de-esterification and cleavage of aromatic ring.



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Characterization flavanone 3β-hydroxylase expressed from Populus euphratica in Escherichia coli and its application in dihydroflavonol production

Abstract

Flavanone 3β-hydroxylase plays very important role in the biosynthesis of flavonoids. A putative flavanone 3β-hydroxylase gene (Pef3h) from Populus euphratica was cloned and over-expressed in Escherichia coli. Induction performed with 0.1 mM IPTG at 20°C led to localization of PeF3H in the soluble fraction. Recombinant enzyme was purified by Ni-NTA affinity. The optimal activity of PeF3H was revealed at pH 7.6 and 35°C. The purified enzyme was stable over pH range of 7.6–8.8 and had a half-life of 1 h at 50°C. The activity of PeF3H was significantly enhanced in the presence of Fe2+ and Fe3+. The K M and V max for the enzyme using naringenin as substrate were 0.23 mM and 0.069 μmoles mg–1min-1, respectively. The K m and V max for eriodictyol were 0.18 mM and 0.013 μmoles mg–1min–1, respectively. The optimal conditions for naringenin bioconversion in dihydrokaempferol were obtained: OD600 of 3.5 for cell concentration, 0.1 mM IPTG, 5 mM α-ketoglutaric acid and 20°C. Under the optimal conditions, naringenin (0.2 g/L) was transformed into 0.18 g/L dihydrokaempferol within 24 h by the recombinant E. coli with a corresponding molar conversion of 88%. Thus, this study provides a promising flavanone 3β-hydroxylase that may be used in biosynthetic applications.



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A novel Bod-mediator biosensor based on Ferrocene and Debaryomyces hansenii yeast cells

Abstract

The mediators ferrocene, 1,1'-dimethylferrocene, ferrocene carboxaldehyde, ferrocene acetonitrile, neutral red, 2,6-dichlorophenolindophenol, thionine, methyl blue, and potassium ferricyanide were used in combination with Debaryomyces hansenii yeast cells to create a biosensor receptor for biochemical oxygen demand (BOD) assays. In this eukaryote, ferrocene and neutral red were observed to efficiently transfer electrons. The biosensor based on ferrocene was characterized by long-term stability (39 days) and a wide range of substrate specificity. The lower detected concentration boundary was 25.2 mg O2/dm3. A high correlation (R = 0.9971) was observed between the results obtained with water samples by this approach and the standard method. This is the first attempt to create a combination of yeast cells and a mediator. The biosensor can be employed for further research on the possibilities of its conventional use.



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Aromatic and monoterpene alcohol accumulation by Eremothecium ashbyi strains differing in riboflavinogenesis

Abstract

We examined the accumulation of phenylethanol, geraniol, citronellol, and nerol by Eremothecium ashbyi Guillermond 1935 strains characterized by different levels of riboflavin synthesis. There was a significant positive correlation between riboflavin and monoterpene alcohol biosyntheses (Spearman's correlation coefficients = 0.81–1.00, p ≤ 0.05). Strain accumulation of the main secondary metabolites such as vitamin B2 and aroma forming compounds was found to be accompanied with an increase in the lipid droplet quantities and the vacuole filling with lipophilic compounds. These phenomena may be used as an indirect measure of riboflavinogenesis intensity and essential oil synthesis.



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Strains of Bacillus ssp. regulate wheat resistance to Septoria nodorum Berk.

Abstract

The ability of Bacillus subtilis Cohn and Bacillus thuringiensis Berliner to induce systemic resistance in wheat plants to the casual agent of Septoria nodorum Berk., blotch has been studied. It has been shown that strains of Bacillus ssp. that possess the capacity for endophytic survival have antagonistic activity against this pathogen in vitro. A reduction of the degree of Septoria nodorum blotch development on wheat leaves under the influence of Bacillus spp. was accompanied by the suppression of catalase activity, an increase in peroxidase activity and H2O2 content, and expression of defence related genes such us PR-1, PR-6, and PR-9. It has been shown that B. subtilis 26 D induces expression levels of wheat pathogenesis-related (PR) genes which marks a SA-dependent pathway of sustainable development and that B. thuringiensis V-5689 and V-6066 induces a JA/ET-dependent pathway. These results suggest that these strain Bacillus spp. promotes the formation of wheat plant resistance to S. nodorum through systemic activation of the plant defense system. The designed bacterial consortium formed a complex biological response in wheat plants infected phytopathogen.



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Implantationen unter Dabigatran-Therapie



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Mitral Stenosis

Description

A previously healthy 29-year-old Mexican woman presented to an emergency department with transient hemiparaesthesias and dysarthria. There was no evidence of stroke on cross-sectional imaging of the head, and she was discharged without a clear diagnosis. Two days later, she returned with acute abdominal pain. Abdominal imaging revealed complete occlusion of the right renal artery, prompting emergency embolectomy. Following the procedure, she developed acute haemoptysis, dyspnoea and hypoxaemia. Chest imaging demonstrated evidence of pulmonary venous hypertension. Cardiac auscultation revealed an opening snap followed by a diastolic murmur with presystolic accentuation. These sounds were better appreciated in combination with phonocardiography, a technique supplanted by echocardiography in the 1970s1 that visualised heart sounds (video 1). An echocardiogram confirmed the presence of mitral stenosis (MS), unifying the syndrome of embolic phenomena, haemoptysis and pulmonary hypertension. She underwent successful mitral valve replacement and has since returned to normal...



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Uncommon presentation of adult-form scimitar syndrome associated with single left pulmonary vein in a pregnant woman

Scimitar syndrome is the constellation of malformations including an abnormal venous drainage of the right lung into the inferior vena cava, associated with the right lung and systemic supply to the right lung. The anomalous vein looks like the curved, Turkish sword (scimitar), hence the name.

The adult form of scimitar syndrome is rare, and it is usually an incidental diagnosis based on the characteristic finding on radiological imaging since the patients are usually asymptomatic or with minimal symptoms.

Our patient presented with a rare presentation of scimitar syndrome, which is tachyarrhythmia (sinus tachycardia, with episodes of supraventricular tachycardia). The diagnosis of scimitar syndrome was made based on the typical radiological finding of the anomalous venous drainage on CT angiography. Our patient does not have the full spectrum of the scimitar syndrome; therefore, she did not suffer from the usual complication (pulmonary hypertension). She was treated with ablation without surgical intervention.



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Mitral valve vegetation diagnosed with oesophageal ultrasound with bronchoscope (EUS-B)

Oesophageal ultrasound with bronchoscope (EUS-B) is designed to evaluate mediastinal structures. We describe a case of a 78-year-old woman who presented with altered mental status for 2 weeks. CT head revealed a subacute infarct in the right middle cerebral artery distribution. She was also found to have a lung mass on chest imaging. EUS-B-guided fine needle aspiration demonstrated the presence of adenocarcinoma in station 7 lymph node and in the mass. Immunohistochemistry confirmed it to be a lung primary as the Thyroid Transcription Factor-1 (TTF-1) was strongly positive. During the procedure, the cardiac valves were evaluated, and a mitral valve vegetation was noted. Formal echocardiography confirmed the presence of the vegetation. During hospital stay, the patient developed fever. Her blood cultures grew oxacillin-resistant Staphylococcus aureus. She was subsequently treated for infective endocarditis. We suggest that the use of EUS-B to routinely scan adjacent structures during a procedure may help obtain additional clinical information that may be critical to patient management.



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Double hit lymphoma presenting as haemophagocytic lymphohistiocytosis

Description

A 67-year-old woman with history of severe rheumatoid arthritis and use of multiple biologics including infliximab, tocilizumab and abatacept presented with fever of 39.1°C and severe pancytopenia (white blood cell count (WBC)=1.0x109/L,  absolute neutrophil count (ANC)=0.55x109/L, haemoglobin=8.7 g/dL, platelets=46x109/L). As part of the pancytopenia evaluation, imaging (CT of the chest, abdomen and pelvis) showed diffuse lymphadenopathy. Further evaluation revealed an elevated ferritin (8564 ng/mL), hypofibrinogenaemia (fibrinogen=95 mg/dL), elevated triglycerides (399 mg/dL) and a soluble interleukin 2 receptor level of 41 167 units/mL, satisfying diagnostic criteria for haemophagocytic lymphohistiocytosis (HLH). A subsequent bone marrow biopsy also revealed morphological evidence of haemophagocytosis (figure 1A), in addition to a population of very large and atypical mononuclear cells with markedly irregular, folded nuclear contours, prominent nucleoli and moderate amounts of cytoplasm (figure 1B,C). A similar large cell infiltrate was identified in the left axillary lymph node, causing complete effacement of nodal architecture (figure...



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Anaemia and respiratory failure in a child: can it be idiopathic pulmonary haemosiderosis?

We present an 8-year-old male child admitted with cough and high-grade fever for 7 days and respiratory difficulty for 2 days. There was a history of blood transfusion at 2 years of age during a respiratory illness. The child was anaemic, tachycardic, tachypnoeic and hypoxic at presentation. Chest examination revealed equal air entry with fine crackles bilaterally. Blood reports were suggestive of anaemia (haemoglobin 6.5 g/dL), leucocytosis and high C reactive protein levels. Chest radiograph revealed bilateral air space opacities involving diffuse lung fields, right more than left. Relevant microbiological workup was negative. Based on the clinical scenario and investigations, a provisional diagnosis of pulmonary haemosiderosis was kept. The patient was started on intravenous pulse methylprednisolone. Fibre-optic bronchoscopy was done following recovery from the acute event. Bronchoalveolar lavage demonstrated a significant number of haemosiderin-laden macrophages confirming pulmonary haemosiderosis.



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A rare case of thrombotic microangiopathy triggered by acute pancreatitis

Thrombotic microangiopathy (TMA) occurring after acute pancreatitis is rarely described. Without prompt intervention, TMA can be, and often is, lethal, so prompt recognition is important. Here, we present a case of a 61-year-old woman with a history of alcohol misuse who presented with epigastric pain, nausea and vomiting after binge drinking. Elevated serum lipase and imaging were suggestive of acute-on-chronic pancreatitis. Although the patient’s symptoms of acute pancreatitis subsided, her anaemia, thrombocytopenia and acute kidney injury worsened. A peripheral blood smear revealed schistocytes, prompting suspicion for TMA. Therapeutic plasma exchange (TPE) was promptly initiated and she completed 10 TPE sessions that improved her anaemia and serum creatinine and resolved the thrombocytopenia. Since TPE was effective and the ADAMTS13 assay revealed 55% activity in the absence of anti-ADAMTS13 IgG prior to initiation of therapy, a confident diagnosis of TMA caused by acute pancreatitis was made. There was no evidence of relapse 2 years later.



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Novel use of combination of electromyography and ultrasound to guide quadratus lumborum block after open appendicectomy

The quadratus lumborum (QL) block facilitates the administration of anaesthesia to the anterior abdominal wall. The use of ultrasound (US) improves the accuracy of the QL block and reduces the risk of adverse events. Electromyography (EMG) in combination with US for muscle plane blocks has not been described previously. We postulated that the addition of EMG-guided needle positioning might assist the execution of this block. This case report describes the first use of combined needle EMG and US to carry out a QL block performed for postoperative analgesia following an open appendicectomy.



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Pericardial incidentaloma: benign pericardial cyst

Description

A 73-year-old woman with hypertension and atrial fibrillation presented with head and neck injury after mechanical fall. During workup, chest X-ray anteroposterior view (figure 1) revealed a rounded opacity silhouetting the left heart border and hilum. Subsequent contrast-enhanced CT of the chest showed single, 6.4 cm, rounded, well-defined, thin-walled, non-enhanced, low attenuated (–20 and 20 Hounsfield Unit) and homogenous cyst-like structure at the left mediastinum connected to pericardial recesses and not attached to adjacent structures (figure 2A–C). Transthoracic echocardiogram ruled out left ventricular aneurysm, aortic aneurysm, solid tumour and outflow tracts obstruction. Although bronchogenic cyst, oesophageal duplication cyst, thymic tumour and mediastinal lymphoma were considered as possible differentials, radiological features such as CT appearance, homogenous attenuation, unrelated to the underlying structures favoured pericardial cyst. Since patient was asymptomatic, patient and family member were unwilling to undergo surgical removal and pathological confirmation. Follow-up with non-enhanced CT of...



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When kinesthetic information is neglected in learning a Novel bimanual rhythmic coordination

Abstract

Many studies have shown that rhythmic interlimb coordination involves perception of the coupled limb movements, and different sensory modalities can be used. Using visual displays to inform the coupled bimanual movement, novel bimanual coordination patterns can be learned with practice. A recent study showed that similar learning occurred without vision when a coach provided manual guidance during practice. The information provided via the two different modalities may be same (amodal) or different (modality specific). If it is different, then learning with both is a dual task, and one source of information might be used in preference to the other in performing the task when both are available. In the current study, participants learned a novel 90° bimanual coordination pattern without or with visual information in addition to kinesthesis. In posttest, all participants were tested without and with visual information in addition to kinesthesis. When tested with visual information, all participants exhibited performance that was significantly improved by practice. When tested without visual information, participants who practiced using only kinesthetic information showed improvement, but those who practiced with visual information in addition showed remarkably less improvement. The results indicate that (1) the information is not amodal, (2) use of a single type of information was preferred, and (3) the preferred information was visual. We also hypothesized that older participants might be more likely to acquire dual task performance given their greater experience of the two sensory modes in combination, but results were replicated with both 20- and 50-year-olds.



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Comparing visual search and eye movements in bilinguals and monolinguals

Abstract

Recent research has suggested that bilinguals show advantages over monolinguals in visual search tasks, although these findings have been derived from global behavioral measures of accuracy and response times. In the present study we sought to explore the bilingual advantage by using more sensitive eyetracking techniques across three visual search experiments. These spatially and temporally fine-grained measures allowed us to carefully investigate any nuanced attentional differences between bilinguals and monolinguals. Bilingual and monolingual participants completed visual search tasks that varied in difficulty. The experiments required participants to make careful discriminations in order to detect target Landolt Cs among similar distractors. In Experiment 1, participants performed both feature and conjunction search. In Experiments 2 and 3, participants performed visual search while making different types of speeded discriminations, after either locating the target or mentally updating a constantly changing target. The results across all experiments revealed that bilinguals and monolinguals were equally efficient at guiding attention and generating responses. These findings suggest that the bilingual advantage does not reflect a general benefit in attentional guidance, but could reflect more efficient guidance only under specific task demands.



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Improving the temperature characteristics and catalytic efficiency of a mesophilic xylanase from Aspergillus oryzae, AoXyn11A, by iterative mutagenesis based on in silico design

To improve the temperature characteristics and catalytic efficiency of a glycoside hydrolase family (GHF) 11 xylanase from Aspergillus oryzae (AoXyn11A), its variants were predicted based on in silico design. Fir...

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Application of response surface methodology for optimization of medium components for the production of secondary metabolites by Streptomyces diastatochromogenes KX852460

A bioactive strain Streptomyces diastatochromogenes KX852460 was selected for the production of secondary metabolites to control the target spot disease on tobacco leaves, caused by the Rhizoctonia solani AG-3. P...

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Human neutrophil elastase induces endothelial cell apoptosis by activating the PERK-CHOP branch of the unfolded protein response [Research]

Human neutrophil elastase impacts on atherosclerotic plaque stability by inducing apoptosis in endothelial cells. Our aim was to investigate the proapoptotic mechanism of elastase on endothelial cells and to evaluate the presence of elastase in human plaque material. Human endothelial cells were treated with purified human neutrophil elastase. Apoptosis was assayed by capsase-3/7 activation, TUNEL, and sub-G1 assay. Activation of unfolded protein response (UPR) effector molecules binding Ig protein, soluble X-binding protein-1, protein kinase RNA-like ER kinase (PERK), and C/EBP-homologous protein (CHOP) was analyzed by RT-PCR, immunocytochemistry, and Western blot. Genetic silencing of CHOP was achieved by small interfering RNA. Elastase induces autophagic–apoptotic forms of endothelial cell death in a time- and dose-dependent manner, in conjunction with a significant increase in phosphorylation/expression of the canonical UPR-activation markers PERK and CHOP. By using CHOP knockdown, we identified CHOP as a key mediator of elastase-induced endothelial cell death. Immunohistochemical analysis of human rupture-prone plaque specimens confirmed the presence of elastase and colocalization with apoptosis. We have demonstrated for the first time that the PERK-CHOP branch of the UPR is causally involved in elastase-induced apoptosis of endothelial cells. Ex vivo analysis of human rupture-prone plaques confirmed the presence of elastase and its colocalization with markers of apoptosis. This novel role of elastase underlines the potential of combined targeting of elastase and endoplasmic reticulum stress in the prevention of plaque progression and cardiovascular events.—Grechowa, I., Horke, S., Wallrath, A., Vahl, C.-F., Dorweiler, B. Human neutrophil elastase induces endothelial cell apoptosis by activating the PERK-CHOP branch of the unfolded protein response.



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Nutritional interventions and the IL-6 response to exercise [Review]

IL-6 is a pleiotropic cytokine with a wide range of biologic effects. In response to prolonged exercise, IL-6 is synthesized by contracting skeletal muscle and released into circulation. Circulating IL-6 is thought to maintain energy status during exercise by acting as an energy sensor for contracting muscle and stimulating glucose production. If tissue damage occurs, immune cells infiltrate and secrete cytokines, including IL-6, to repair skeletal muscle damage. With adequate rest and nutrition, the IL-6 response to exercise is attenuated as skeletal muscle adapts to training. However, sustained elevations in IL-6 due to repeated bouts of unaccustomed activities or prolonged exercise with limited rest may result in untoward physiologic effects, such as accelerated muscle proteolysis and diminished nutrient absorption, and may impair normal adaptive responses to training. Recent intervention studies have explored the role of mixed meals or carbohydrate, protein, -3 fatty acid, or antioxidant supplementation in mitigating exercise-induced increases in IL-6. Emerging evidence suggests that sufficient energy intake before exercise is an important factor in attenuating exercise-induced IL-6 by maintaining muscle glycogen. We detail various nutritional interventions that may affect the IL-6 response to exercise in healthy human adults and provide recommendations for future research exploring the role of IL-6 in the adaptive response to exercise.—Hennigar, S. R., McClung, J. P., Pasiakos, S. M. Nutritional interventions and the IL-6 response to exercise.



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Do aspirin and other NSAIDs confer a survival benefit in men diagnosed with prostate cancer? A pooled analysis of NIH-AARP and PLCO cohorts

<span style="margin: 0px; font-family: 'New times rome',serif;">Prostate cancer is one of the leading causes of cancer death in US men. There is an unmet need to identify modifiable risk factors for prostate cancer survival. Experimental studies have suggested that nonsteroidal anti-inflammatory drugs (NSAIDs) may improve prostate cancer survival through anti-thrombotic and anti-inflammation mechanisms. Results from previous observational studies have been equivocal, and few have assessed whether an etiologically relevant time window of exposure exists. We sampled prostate cancer cases from two large US prospective cohorts-NIH-AARP Diet and Health Study and PLCO Cancer Screening Trial-to investigate whether pre- and post-diagnostic aspirin and non-aspirin NSAID use were associated with prostate cancer-specific and all-cause mortality. Cox proportional hazards regression models estimated hazard ratios (HRs) and 95% confidence intervals (CIs). Study-specific results were meta-analyzed using fixed-effects models. Pre- and post-diagnostic aspirin or non-aspirin NSAID use were not statistically significantly associated with prostate cancer-specific mortality. However, occasional (less than daily) and daily aspirin users five years or more before prostate cancer diagnosis had 18% (HR=0.82; 95%CI=0.75 to 0.90) and 15% (HR=0.85; 95%CI=0.77 to 0.94) reduced all-cause mortality versus nonusers. Similarly, post-diagnostic occasional and daily aspirin use were associated with 17% (HR=0.83; 95%CI=0.72 to 0.95) and 25% (HR=0.75; 95%CI=0.66 to 0.86) reduced all-cause mortality, independent of pre-diagnostic aspirin use. This study suggests that aspirin or non-aspirin NSAIDs are not associated with prostate cancer survival. However, aspirin use both before and after prostate cancer diagnosis was associated with longer overall survival, highlighting the importance of comorbidity prevention among prostate cancer survivors. </span>



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Yokenella regensburgei Septicemia in a Chinese Farmer Immunosuppressed by HIV: A Case Report and Literature Review

Yokenella regensburgei is a member in the family Enterobacteriaceae and a few cases have been reported in immunocompromised hosts. Herein, we described a case of septicemia in a human immunodeficiency virus (HIV) infected patient in South West China, which is the first reported case of Y. regensburgei infection in HIV-infected populations. We then reviewed the literature on all the reported cases of Y. regensburgei infection worldwide and presented some common features of them. Our case report and literature review will help increase the knowledge of the bacterium Y. regensburgei and its clinical implications.

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Advanced Functional Tumor Imaging and Precision Nuclear Medicine Enabled by Digital PET Technologies

The purpose of this article is to provide a brief overview of the background, basic principles, technological evolution, clinical capabilities, and future directions for functional tumor imaging as PET evolves from the conventional photomultiplier tube-based platform into a fully digital detector acquisition platform. The recent introduction of solid-state digital photon counting PET detector is the latest evolution of clinical PET which enables faster time-of-flight timing resolution that leads to more precise localization of the annihilation events and further contributes to reduction in partial volume and thus makes high definition and ultrahigh definition PET imaging feasible with current standard acquisition procedures. The technological advances of digital PET can be further leveraged by optimizing many of the acquisition and reconstruction methodologies to achieve faster image acquisition to improve cancer patient throughput, lower patient dose in accordance with ALARA, and improved quantitative accuracy to enable biomarker capability. Digital PET technology will advance molecular imaging capabilities beyond oncology and enable Precision Nuclear Medicine.

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Performance Analysis of Photovoltaic Water Heating System

Performance of solar photovoltaic water heating systems with direct coupling of PV array to DC resistive heating elements has been studied and compared with solar photothermal systems. An analysis of optimum fixed load resistance for different climate conditions has been performed for simple PV heating systems. The optimum value of the fixed load resistance depends on the climate, especially on annual solar irradiation level. Use of maximum power point tracking compared to fixed optimized load resistance increases the annual yield by 20 to 35%. While total annual efficiency of the PV water heating systems in Europe ranges from 10% for PV systems without MPP tracking up to 15% for system with advanced MPP trackers, the efficiency of solar photothermal system for identical hot water load and climate conditions is more than 3 times higher.

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Formulation and Development of a Validated UV-Spectrophotometric Analytical Method of Rutin Tablet

Rutin is available in some foods, fruits, and vegetables. It has various beneficial medical effects making it useful in the treatment of various diseases. Rutin is available in different oral dosage forms such as tablets or capsules, widely available in the market. Rutin and many herbal medicines lack quality control due to unavailability of analytical methods. In this study, we formulated rutin tablet and studied its stability using a simple developed analytical method. The dissolution profile of our formulated tablet was also inspected. The results showed that our developed method was linear (), precise (% RSD = 0.026), and accurate (% recovery = 98.55–103.34). The formulated rutin tablet was stable under accelerated conditions as well as room temperature for 150 days (% assay > 91.69). The dissolution profile over 45 minutes of our formulated tablet showed a better dissolution (26.5%) compared with the internationally marketed Rutin® tablet (18.5%). This study can serve as a guideline to companies that manufacture herbal products to improve their formulated herbs and apply validated analytical methods to check the quality of their product.

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Malignant Peripheral Nerve Sheath Tumors State of the Science: Leveraging Clinical and Biological Insights into Effective Therapies

Malignant peripheral nerve sheath tumor (MPNST) is the leading cause of mortality in patients with neurofibromatosis type 1. In 2002, an MPNST consensus statement reviewed the current knowledge and provided guidance for the diagnosis and management of MPNST. Although the improvement in clinical outcome has not changed, substantial progress has been made in understanding the natural history and biology of MPNST through imaging and genomic advances since 2002. Genetically engineered mouse models that develop MPNST spontaneously have greatly facilitated preclinical evaluation of novel drugs for translation into clinical trials led by consortia efforts. Continued work in identifying alterations that contribute to the transformation, progression, and metastasis of MPNST coupled with longitudinal follow-up, biobanking, and data sharing is needed to develop prognostic biomarkers and effective prevention and therapeutic strategies for MPNST.

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Endoscopic Submucosal Dissection of Gastric Epithelial Neoplasms after Partial Gastrectomy: A Single-Center Experience

Aims. To investigate the feasibility and safety of endoscopic submucosal dissection (ESD) of gastric epithelial neoplasms in the remnant stomach (GEN-RS) after various types of partial gastrectomy. Methods. This study included 29 patients (31 lesions) who underwent ESD for GEN-RS between March 2006 and August 2016. Clinicopathologic data were retrieved retrospectively to assess the therapeutic ESD outcomes, including en bloc and complete resection rates and procedure-related adverse events. Results. The en bloc, complete, and curative resection rates were 90%, 77%, and 71%, respectively. The types of previous gastrectomy, tumor size, macroscopic type, and tumor histology were not associated with incomplete resection. Only tumors involving the suture lines from the prior partial gastrectomy were significantly associated with incomplete resection. The procedure-related bleeding and perforation rates were 6% and 3%, respectively; none of the adverse events required surgical intervention. During a median follow-up period of 25 months (range, 6–58 months), there was no recurrence in any case. Conclusions. ESD is a safe and feasible treatment for GEN-RS regardless of the previous gastrectomy type. However, the complete resection rate decreases for lesions involving the suture lines.

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Herbal Medicine Treatment for Children with Autism Spectrum Disorder: A Systematic Review

Objective. To summarize and evaluate the efficacy and safety of herbal medicines used for the treatment of autism spectrum disorder (ASD) in children. Methods. Thirteen electronic databases were searched from their inception to November 2016. Randomized controlled trials (RCTs) that assessed the efficacy of herbal medicines alone or in combination with other Traditional Chinese Medicine treatments for ASD in children were included. The Cochrane Risk of Bias Tool was used and other data analyses were performed using RevMan (Version 5.3). Results. Ten RCTs involving 567 patients with ASD were included for qualitative synthesis. In conjunction with conventional therapy, herbal medicines significantly improved the Childhood Autism Rating Scale (CARS) score, but the results of effects on total effective rate (TER) were different between the included studies. The use of herbal medicines with integrative therapy improved the CARS score and TER. In the studies that documented adverse events, no serious events were associated with herbal medicines. Conclusions. The efficacy of herbal medicines for the treatment of ASD appears to be encouraging but was inconclusive owing to low methodological quality, herbal medicine diversity, and small sample size of the examined studies.

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The association of low penetrance genetic risk modifiers with colorectal cancer in lynch syndrome patients: a systematic review and meta-analysis

Abstract

Lynch syndrome (LS) is a highly penetrant inherited cancer predisposition syndrome accounting for approximately 1000 cases of colorectal cancer (CRC) in the UK annually. LS is characterised by autosomal dominant inheritance and germline mutations in DNA mismatch repair genes. The penetrance is highly variable and the reasons for this have not been fully elucidated. This study investigates whether low penetrance genetic risk factors may result in phenotype modification in LS patients. To conduct a systematic literature review and meta-analysis to assess the association between low penetrance genetic risk modifiers and CRC in LS patients. A systematic review was conducted of the PubMed and HuGENet databases. Eligibility of studies was determined by pre-defined criteria. Included studies were analysed via the per-allele model and assessed by pooled odds ratios and establishing 95% confidence intervals. Study heterogeneity was assessed via Cochrane's Q statistic and I2 values. Publication bias was evaluated with funnel plots. Subgroup analysis was conducted on gender. Statistical software used was the Metafor package for the R programme version 3.1.3. Sixty-four polymorphisms were identified and sufficient data was available for analysis of ten polymorphisms, with between 279 and 1768 CRC cases per polymorphism. None demonstrated association with CRC risk in LS patients. However in sub-group analysis the polymorphism rs16892766 (8q23.3) was significant in males (OR 1.53, 95% CI 1.12–2.10). The variable phenotype presentation of the disease still remains largely unexplained, and further investigation is warranted. Other factors may also be influencing the high variability of the disease, such as environmental factors, copy number variants and epigenetic alterations. Investigation into these areas is needed as well as larger and more definitive studies of the polymorphisms analysed in this study.



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Hyaluronan Nanoparticles Selectively Target Plaque-Associated Macrophages and Improve Plaque Stability in Atherosclerosis

TOC Graphic

ACS Nano
DOI: 10.1021/acsnano.7b01385
ancac3?d=yIl2AUoC8zA


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Single-Atom Scale Structural Selectivity in Te Nanowires Encapsulated Inside Ultranarrow, Single-Walled Carbon Nanotubes

TOC Graphic

ACS Nano
DOI: 10.1021/acsnano.7b02225
ancac3?d=yIl2AUoC8zA


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Nicotinic acid phosphoribosyltransferase regulates cancer cell metabolism, susceptibility to NAMPT inhibitors and DNA repair

In the last decade, substantial efforts have been made to identify NAD+ biosynthesis inhibitors, specifically against nicotinamide phosphoribosyltransferase (NAMPT), as preclinical studies indicate their potential efficacy as cancer drugs. However, the clinical activity of NAMPT inhibitors has proven limited, suggesting that alternative NAD+ production routes exploited by tumors confer resistance. Here we show the gene encoding nicotinic acid phosphoribosyltransferase (NAPRT), a second NAD+ producing enzyme, is amplified and overexpressed in a subset of common types of cancer, including ovarian cancer, where NAPRT expression correlates with a BRCAness gene expression signature. Both NAPRT and NAMPT increased intracellular NAD+ levels. NAPRT silencing reduced energy status, protein synthesis, and cell size in ovarian and pancreatic cancer cells. NAPRT silencing sensitized cells to NAMPT inhibitors both in vitro and in vivo; similar results were obtained with the NAPRT inhibitor 2-hydroxynicotinic acid. Reducing NAPRT levels in a BRCA2-deficient cancer cell line exacerbated DNA damage in response to chemotherapeutics. In conclusion, NAPRT-dependent NAD+ biosynthesis contributes to cell metabolism and to the DNA repair process in a subset of tumors. This knowledge could be used to increase the efficacy of NAMPT inhibitors and chemotherapy.

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A new l-amino acid oxidase from Bothrops jararacussu snake venom: Isolation, partial characterization, and assessment of pro-apoptotic and antiprotozoal activities

S01418130.gif

Publication date: October 2017
Source:International Journal of Biological Macromolecules, Volume 103
Author(s): Sante E.I. Carone, Tássia R. Costa, Sandra M. Burin, Adélia C.O. Cintra, Karina F. Zoccal, Francine J. Bianchini, Luiz F.F. Tucci, João J. Franco, Maria R. Torqueti, Lúcia H. Faccioli, Sérgio de Albuquerque, Fabíola A. de Castro, Suely V. Sampaio
A new l-amino acid oxidase (LAAO) from Bothrops jararacussu venom (BjussuLAAO-II) was isolated by using a three-step chromatographic procedure based on molecular exclusion, hydrophobicity, and affinity. BjussuLAAO-II is an acidic enzyme with pI=3.9 and molecular mass=60.36kDa that represents 0.3% of the venom proteins and exhibits high enzymatic activity (4884.53U/mg/mim). We determined part of the primary sequence of BjussuLAAO-II by identifying 96 amino acids, from which 34 compose the N-terminal of the enzyme (ADDRNPLEECFRETDYEEFLEIARNGLSDTDNPK). Multiple alignment of the partial BjussuLAAO-II sequence with LAAOs deposited in the NCBI database revealed high similarity (95–97%) with other LAAOs isolated from Bothrops snake venoms. BjussuLAAO-II exerted a strong antiprotozoal effect against Leishmania amazonensis (IC50=4.56μg/mL) and Trypanosoma cruzi (IC50=4.85μg/mL). This toxin also induced cytotoxicity (IC50=1.80μg/mL) and apoptosis in MCF7 cells (a human breast adenocarcinoma cell line) by activating the intrinsic and extrinsic apoptosis pathways, but were not cytotoxic towards MCF10A cells (a non-tumorigenic human breast epithelial cell line). The results reported herein add important knowledge to the field of Toxinology, especially for the development of new therapeutic agents.



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Microgramma vacciniifolia (Polypodiaceae) fronds contain a multifunctional lectin with immunomodulatory properties on human cells

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Publication date: October 2017
Source:International Journal of Biological Macromolecules, Volume 103
Author(s): Leydianne Leite de Siqueira Patriota, Thamara Figueiredo Procópio, Jéssica de Santana Brito, Virginie Sebag, Ana Patrícia Silva de Oliveira, Ana Karine de Araújo Soares, Leyllane Rafael Moreira, Thâmarah de Albuquerque Lima, Tatiana Soares, Túlio Diego da Silva, Patrícia Maria Guedes Paiva, Virgínia Maria Barros de Lorena, Cristiane Moutinho Lagos de Melo, Lidiane Pereira de Albuquerque, Thiago Henrique Napoleão
In this study, we report the purification and characterization of a multifunctional lectin (MvFL) from Microgramma vacciniifolia fronds as well as its immunomodulatory properties on human peripheral blood mononuclear cells (PBMCs). MvFL (pI 4.51; 54kDa) is a glycoprotein able to inhibit trypsin activity and that has sequence similarities (32% coverage) with a plant RNA-binding protein. Hemagglutinating activity of MvFL was not altered by heating at 100°C for 30min, but was reduced in alkaline pH (8.0 and 9.0). Fluorimetric analyses showed that this lectin did not undergo marked conformational changes when heated. However, the MvFL conformation changed depending on the pH. MvFL at 6.25–25μg/mL was not cytotoxic to lymphocytes present among PBMCs. The PBMCs incubated for 24h with the lectin (12.5μg/mL) showed increased TNF-α, IFN-γ, IL-6, IL-10, and nitric oxide production. MvFL also stimulated T lymphocytes from PBMCs to differentiate into CD8+ cells. The activation (indicated by CD28 expression) of these cells was also stimulated. In conclusion, MvFL is a heat-stable and multifunctional protein, with both lectin and trypsin inhibitor activities, and capable of inducing predominantly a Th1 response in human PBMCs as well as activation and differentiation of T lymphocytes.



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Molecular characterization and application of lipase from Bacillus sp. PU1 and investigation of structural changes based on pH and temperature using MD simulation

S01418130.gif

Publication date: October 2017
Source:International Journal of Biological Macromolecules, Volume 103
Author(s): Palanichamy Esakkiraj, Christian Bharathi Antonyraj, Balraj Meleppat, Dasari Ankaiah, Repally Ayyanna, Syed Ibrahim Basheer Ahamed, Venkatesan Arul
A gene coding lipase from Bacillus sp. PU1 was cloned and expressed in E. coli BL21(DE3) pLysS. The purified lipase has a molecular weight of 23kDa, is highly alkaline (pH range 8–10) and mesophilic (20–50°C). Three dimensional structure of the lipase was modeled by comparative homology and identified as a typical serine lipase by the presence of conserved Ser77, Asp133, His156. The molecular stability and behavior of the lipase was carried out using MD simulation studies at different pH and temperature was performed in comparison with biochemical analysis. Structural modifications of the lipase under these conditions were trapped by dihedral based FEL analysis and the functional loops (loop-H5/B4 and loop-H6/B5 of lipase) are identified which would cause the catalytic behavior of the lipase by high flexibility. Further characteristic feature of lipase are observed as follows; SDS completely inhibits the lipase activity and enzyme activity is enhanced with non-ionic surfactants. The lipase was highly stable in different organic solvents and also it could tolerate NaCl (0.4–0.8M). This enzyme was found to disrupt the biofilm of tested pathogenic bacterial strains.



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Human Myosin VIIa Is a Very Slow Processive Motor Protein on Various Cellular Actin Structures [Molecular Bases of Disease]

Human myosin VIIa (MYO7A) is an actin-linked motor protein associated with human Usher syndrome (USH) type 1B, which causes human congenital hearing and visual loss. While it has been thought that the role of human myosin VIIa is critical for USH1 protein tethering with actin and transportation along actin bundles in inner-ear hair cells, myosin VIIa′s motor function remains unclear. Here, we studied the motor function of the tail-truncated human myosin VIIa dimer (HM7AΔTail/LZ) at the single-molecule level. We found that the HM7AΔTail/LZ moves processively on single actin filaments with a step size of 35 nm. Dwell-time distribution analysis indicated an average waiting time of 3.4 s, yielding ≈0.3 s-1 for the mechanical turnover rate, hence, the velocity of HM7AΔTail/LZ was extremely slow, at 11 nm s-1. We also examined HM7AΔTail/LZ movement on various actin structures in demembranated cells. HM7AΔTail/LZ showed unidirectional movement on actin structures at cell edges, such as lamellipodia and filopodia. However, HM7AΔTail/LZ frequently missed steps on actin tracks and exhibited bi-directional movement at stress fibers, which was not observed with tail-truncated myosin Va. These results suggest that the movement of the human myosin VIIa motor protein is more efficient on lamellipodial and filopodial actin tracks than on stress fibers, which are composed of actin filaments with different polarity, and that the actin structures influence the characteristics of cargo transportation by human myosin VIIa. In conclusion, myosin VIIa movement appears to be suitable for translocating USH1 on stereocilia actin bundles in inner-ear hair cells.

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Carbon dioxide-dependent regulation of NF-{kappa}B family members RelB and p100 gives molecular insight into CO2-dependent immune regulation [Molecular Bases of Disease]

CO2 is a physiological gas normally produced in the body during aerobic respiration. Hypercapnia (elevated blood pCO2 > ≈ 50mmHg) is a feature of several lung pathologies e.g. chronic obstructive pulmonary disease (COPD). Hypercapnia is associated with increased susceptibility to bacterial infections and suppression of inflammatory signaling. The NF-κB pathway has been implicated in these effects, however, the molecular mechanisms underpinning cellular sensitivity of the NF-κB pathway to CO2 is not fully elucidated. Here we identify several novel CO2-dependent changes in the NF-κB pathway. NF-κB family members p100 and RelB translocate to the nucleus in response to CO2. A cohort of RelB protein-protein interactions (e.g. with Raf-1 and IκBα) are altered by CO2 exposure, while others are maintained (e.g. with p100). RelB is processed by CO2 in a manner dependent on a key C-terminal domain located in its transactivation domain. Loss of the RelB transactivation domain alters NF-κB -dependent transcriptional activity and loss of p100 alters sensitivity of RelB to CO2. Thus, we provide molecular insight into the CO2-sensitivity of the NF-κB pathway and implicate altered RelB/p100-dependent signaling in the CO2-dependent regulation of inflammatory signaling.

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Cargo engagement protects protease adaptors from degradation in a substrate-specific manner [Microbiology]

Protein degradation in bacteria is a highly controlled process involving proteolytic adaptors that regulate protein degradation during cell cycle progression or during stress responses. Many adaptors work as scaffolds that selectively bind cargo and tether substrates to their cognate proteases to promote substrate destruction, while others primarily activate the target protease. Because adaptors must bind their cognate protease, all adaptors run the risk of being recognized by the protease as substrates themselves, a process that could limit their effectiveness. Here we use purified proteins in a reconstituted system and in vivo studies to show that adaptors of the ClpXP protease are readily degraded but that cargo binding inhibits this degradation. We found that this principle extends across several adaptor systems, including the hierarchical adaptors that drive the Caulobacter bacterial cell cycle and the quality-control adaptor SspB. We also found that the ability of a cargo to protect its adaptor is adaptor-substrate specific, as adaptors with artificial degradation tags were not protected even though cargo binding is unaffected. Our work points to an optimization of inherent adaptor degradation and cargo binding, which ensures that robust adaptor activity is maintained when high amounts of substrate must be delivered and adaptors can be eliminated when their tasks have been completed.

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Gene duplication and neo-functionalization in the evolutionary and functional divergence of metazoan copper transporters Ctr1 and Ctr2 [Metabolism]

Copper is an essential element for proper organismal development and is involved in a range of processes including oxidative phosphorylation, neuropeptide maturation, and connective tissue maturation. The copper transporter (Ctr) family of integral membrane proteins is ubiquitously found in eukaryotes and mediates the high-affinity transport of Cu+ across both the plasma membrane and endomembranes. While mammalian Ctr1 functions as a Cu+ transporter for Cu acquisition and is essential for embryonic development, a homologous protein, Ctr2, has been proposed to function as a low-affinity Cu transporter, a lysosomal Cu exporter, or a regulator of Ctr1 activity, but its functional and evolutionary relationship to Ctr1 is unclear. Here, we report on a biochemical, genetic, and phylogenetic comparison of metazoan Ctr1 and Ctr2, suggesting that Ctr2 arose over 550 million years ago as a result of a gene duplication event followed by a loss of Cu+ transport activity. Using a random mutagenesis and growth selection approach, we identified amino acid substitutions in human and mouse Ctr2 proteins that support Cu-dependent growth in yeast and enhance Cu accumulation in Ctr1-/- mouse embryonic fibroblasts. These mutations revert Ctr2 to a more ancestral Ctr1-like state while maintaining endogenous functions, such as stimulating Ctr1 cleavage. We suggest key structural aspects of metazoan Ctr1 and Ctr2 that discriminate between their biological roles, providing mechanistic insights into the evolutionary, biochemical, and functional relationships between these two related proteins.

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FOXO3/PGC-1{beta} signaling axis is essential for cancer stem cell properties of pancreatic ductal adenocarcinoma [Cell Biology]

In 95% of patients with pancreatic ductal adenocarcinoma (PDAC), recurrence is observed following chemotherapy. Findings from several studies have indicated that cancer stem cells (CSCs) are resistant to anti-cancer agents and may be involved in cancer recurrence and metastasis. The CD44 protein is a major CSC marker, and CD44 also plays an indispensable role in the CSC properties in several cancers, including pancreatic cancer; however, no clinical approach exists to inhibit CD44 activity. Here, we have performed knockin/knock down experiments and we demonstrate that the forkhead box O3 (FOXO3)/liver kinase B1 (LKB1)/AMP-activated protein kinase (AMPK)/PPAR-γ co-activator-1β (PGC- 1β)/pyruvate dehydrogenase -A1 (PDHA1) pathway is essential for CD44 expression and CSC properties. We observed that patients exhibiting high PDHA1 expression have a poor prognosis. Systemic PGC-1β knockout mice are fertile and viable and do not exhibit an overt phenotype under normal conditions. This suggests that cGMP induction and PGC-1β inhibition represent potential strategies for treating patients with PDAC.

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The MicroRNA Machinery Regulates Fasting-Induced Changes in Gene Expression and Longevity in Caenorhabditis elegans [Gene Regulation]

Intermittent fasting (IF) is a dietary restriction regimen that extends the lifespans of Caenorhabditis elegans and mammals by inducing changes in gene expression. However, how IF induces these changes and promotes longevity remains unclear. One proposed mechanism involves gene regulation by microRNAs (miRNAs), small non-coding RNAs (approximately 22 nucleotides) that repress gene expression and whose expression can be altered by fasting. To test this proposition, we examined the role of the miRNA machinery in fasting-induced transcriptional changes and longevity in C. elegans. We revealed that fasting up-regulated the expression of miRNA-induced silencing complex (miRISC) components, including Argonaute and GW182, and of the miRNA-processing enzyme DRSH-1 (the ortholog of the Drosophila Drosha enzyme). Our lifespan measurements demonstrated that IF-induced longevity was suppressed by knockout or knockdown of miRISC components and was completely inhibited by drsh-1 ablation. Remarkably, drsh-1 ablation inhibited the fasting-induced changes in the expression of the target genes of DAF-16, the insulin/IGF-1 signaling effector in C. elegans. Fasting-induced transcriptome alterations were substantially and modestly suppressed in the drsh-1 null mutant and the null mutant of ain-1, a gene encoding GW182, respectively. Moreover, miRNA array analyses revealed that the expression levels of numerous miRNAs changed after 2 days of fasting. These results indicate that components of the miRNA machinery, especially the miRNA-processing enzyme DRSH-1, play an important role in mediating IF-induced longevity via the regulation of fasting-induced changes in gene expression.

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Reconstruction for Skull Base Defect Using Fat-Containing Perifascial Areolar Tissue

imageAbstract: Skull base reconstruction is a challenging task. The method depends on the anatomical complexity and size of the defect. We obtained tissue by harvesting fat-containing perifascial areolar tissue (PAT) for reconstruction of limited skull base defects and volume augmentation. We demonstrated the effective option for reconstruction of limited skull base defects and volume augmentation. From October 2013 to November 2015, 5 patients underwent operations using fat-containing PAT to fill the defect in skull base and/or perform volume replacement in the forehead. Perifascial areolar tissue with 5- to 10-mm fat thickness was harvested from the inguinal region. The fat-containing PAT was grafted to the defect contacting the vascularized wound bed. Patients were followed up in terms of their clinical symptoms and postoperative magnetic resonance imaging findings. Four patients were treated using fat-containing PAT after tumor resection. One patient was treated for a posttraumatic forehead depression deformity. The fat-containing PAT included 5- to 9-mm fat thickness in all cases. The mean size of grafted PAT was 65.6 cm2 (28–140 cm2). The mean follow-up period was 18.6 months (12–31 months). There was no notable complication. There was no donor site morbidity. We can harvest PAT with fat easily and obtain the sufficient volume to treat the defect. It also could be used with other reconstructive method, such as a free flap or a regional flap to fill the left dead space. Therefore, fat-containing PAT could be additional options to reconstruction of skull base defect.

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Vascular Nature and Existence of Anastomoses of Extrinsic Postauricular Fascia: Application for Staged Auricular Reconstruction

imageAbstract: A staged auricular reconstruction in microtia patients was developed by using superficial mastoid fascia (as part of extrinsic postauricular fascia) to cover the cartilagenous framework due to its highly vascularized nature. Three branches of external carotid artery (superficial temporal artery, posterior auricular artery and occipital artery) were found to supply this fascia, this study was therefore aimed to investigate the dimension of blood supply from each vessel and also to demonstrate the existence of anastomoses among these arteries. Thirty-eight pinnas and postauricular fascias from Thai fresh adult cadavers were included to document the anastomoses by showing both perfused dye connection (10 dissections) and visible anastomotic branches (8 dissections) among them. Distribution of each vessel trunk and its branches were demonstrated using superimposed illustration in the other 20 dissections with dye injection into each artery to designate 3 zones of anastomotic area between each arterial pair. Maximal size of viable postauricular fascial flap for staged reconstruction according to this vascular study was thus estimated to be at least 5 cm above and 3 cm below the Frankfurt horizontal plane and about 6 cm posterior to external acoustic meatus owing to the course of posterior auricular artery and its anastomoses. In addition, greater size of flap with dual blood supply from both superficial temporal and posterior auricular arteries can be raised by harvesting beyond 5 cm above external acoustic meatus.

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The Impact of Cervical Musculoskeletal Disorders on UK Consultant Plastic Surgeons: Can We Reduce Morbidity With Applied Ergonomics?

imageBackground: Based on anecdotal and observational evidence, we hypothesized that the prevalence of cervical musculoskeletal disorder (C-MSD) would be high among plastic surgeons. A questionnaire review was undertaken to test this hypothesis. Ergonomic assessment was undertaken to assess causal factors of C-MSD. Method: An anonymous questionnaire recording demographics, physical symptoms and behavioral responses to C-MSD was distributed to UK Plastic Surgery consultants. The postural impact of wearing loupes was assessed using motion capture techniques and recording cervical muscular activity. Results: The questionnaire response rate was 81%. The prevalence of cervical spine morbidity was recorded as 32%. Employment implications included 28% of the cohort requiring sick leave. The professional impact was 7% permanently modifying their practice. There were 2 factors significant for C-MSD, the surgeons' age and the duration in hours of wearing loupes per week. Ergonomic assessment of surgeons operating in loupes demonstrated: 1. increased forward and lateral cervical flexion; 2. increased cervical muscular activity to maintain the protracted “head forward” posture; and 3. prolonged static posturing to maintain head position for visual focus. Table height adjustment and variation of loupe working distance can reduce neck flexion. Conclusions: Cervical morbidity is a prevalent problem among plastic surgeons. Long procedures, static postures and neck flexion result in the “head forward” posture. This posture exaggerates when operating with loupe magnification. Early-middle-aged consultants are more prone to cervical morbidity hence afflicted when at the top of their game. The work force is diminished for a potentially avoidable morbidity. Rather than accept this morbidity, co-operation between plastic surgeons and ergonomist may help to reduce injury.

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Nipple-Sparing Mastectomy Improves Long-Term Nipple But Not Skin Sensation After Breast Reconstruction: Quantification of Long-Term Sensation in Nipple Sparing Versus Non-nipple Sparing Mastectomy

imageBackground: Changes in breast sensation after reconstruction are expected. Return of breast sensation after reconstruction and whether nipple-sparing mastectomy offers a substantial benefit in terms of sensation has been inconsistently documented in the literature. We conducted the current study using the pressure-specified sensory device to quantify postoperative breast sensation in patients undergoing nipple-sparing versus non–nipple-sparing mastectomy. Methods: Consecutive adult women who underwent nipple-sparing (NSM) and non-NSM (NNSM) and were at least 18 months postreconstruction were included. Breast measurements were taken in 4 quadrants (upper/lower lateral, upper/lower medial) and nipple. Averaged skin cutaneous thresholds [(UL+LL+UM+LM)/4] and nipple sensation between NSM and NNSM were compared as the primary outcome measure. A generalized estimating equations model was used; univariate and multivariate variable analyses were done when appropriate. Results: Forty-four patients (74 breasts) were examined (53 NNSM vs 21 NSM). The groups were further subdivided into autologous versus implant-based reconstruction. Averaged cutaneous skin thresholds for quadrants were better for the NSM, 51.8(±24.5) g/mm2 versus NNSM, 56.5(±25.7) g/mm2, although this difference was not statistically significant. However, NSM breasts measured higher nipple or nipple area sensitivity, 44.5(±30.8) g/mm2 versus NNSM, 83.8(±27.4) g/mm2 (P 3) was a predictor of decreased sensation.

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Course Review: Cadaveric Hand Trauma Course

Abstract: The St Thomas' Cadaveric Hand Trauma course is described and evaluated.

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Closure of Meningomyelocele Defects With Sensate Medial Dorsal Intercostal Artery Perforator Flaps

imageNo abstract available

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Restoration of Complicated Epicanthus: Modified Reverse Skin Redraping With Mini-epicanthoplasty for Rescue in Unsatisfied Epicanthoplasty Patients

imagePurpose: We had reported the surgical outcome of reverse skin redraping technique for restoration of previously performed epicanthoplasty. In this study, we introduce a modified reverse skin redraping technique that added mini-epicanthoplasty in patients with unsatisfactory results after epicanthoplasty. Methods: Three hundred twenty-four patients (288 female and 36 male patients) who had unsatisfied results with previous epicanthoplasty and that were treated with our modified restoration surgery were included in this study. Results: The mean preoperative interepicanthal distance was 33.6 mm, and the mean postoperative interepicanthal distance was 36.9 mm; the mean difference in the interepicanthal distance before and after restoration surgery was 3.3 mm. Satisfactory aesthetic results were obtained with improvements in areas of asymmetry, overexposure of the caruncle, and the appearance of the previous scar. Only minor complications developed in 15 patients (4.6%) that were resolved with minor revisions. No severe complications requiring reoperations were noted. Conclusions: Our modified method involving reverse skin redraping and mini-epicanthoplasty is simple and reproducible and is useful for resolution of unsatisfactory results to obtain a naturally shaped epicanthus.

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Individualized Plastic Reconstruction Strategy for Patients With Ectodermal Dysplasia Syndrome

imageAbstract: Ectodermal dysplasia syndrome is a hereditary disease of ectodermal origin. Appearances of nail dystrophy, alopecia or hypotrichosis, saddle nose deformity, and palmoplantar hyperkeratosis are usually associated with a lack of sweat glands as well as partial or complete absence of teeth. These manifestations are usually corrected only with oral rehabilitation by mounting dentures. In this study, plastic rehabilitation was developed to correct the special features of patients with ectodermal dysplasia. Four men and 1 woman with ectodermal dysplasia syndrome were treated. Four patients showed dysostosis of the midface, and rhinoplasty with costal bone was performed, whereas cosmetic operation aiming to repair soft tissue defects was adopted for the last patient. After plastic corrections, all 5 patients were satisfied with the results and had no social embarrassment.

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“Stamp Perforation” Technique for Correction of Prominent Mandibular Angle: 10 Years of Experience in Mandibular Reshaping in Asians

imageAbstract: Resection of a prominent mandibular angle is commonly used in Eastern society to improve the lower one third facial proportion. Historically, this procedure had a high complication rate, such as severe bleeding, asymmetry of the angle reduction, and “second mandibular angle.” A safer and more effective way of performing such procedures is needed. The aim of this study is to introduce 3 instruments, a tunable guide handpiece, milling cutter, and flywheel, which were invented by the author, as well as a related ostectomy technique for correcting prominent mandibular angles using a modified full-thickness marginal ostectomy of the mandibular corpus angle, named the “stamp perforation” technique. This technique has 4 highlights: First, it ensures a smooth symmetric contour. Second, it prevents the risks of rupture of the inferior alveolar vessel and facial artery, ensuring the safety of this approach. Third, the “stamp perforation” technique eases the removal of bone fragments, shortening the operation time. Fourth, the recovery time of patients treated with this approach is much shorter than with the traditional approaches. From January 2006 to January 2016, 1106 patients underwent the surgery to contour the prominent mandible angles, and satisfactory results were achieved. Thus, we recommend the instruments as well as the “stamp perforation” technique for correcting prominent mandibular angles, and we hope that our 10 years of experience could provide a reference for other plastic surgeons.

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Pedicled Flap Closure as an Adjunct for Infected Ventricular Assist Devices

imagePurpose: End-stage cardiac disease has resulted in an increased utilization of cardiac transplantation or long-term mechanical assistance to sustain life. Though left ventricular assist devices (LVAD) have revolutionized the treatment algorithm for these patients, these devices carry a substantial infection rate, ranging from 30% to 50%. We report our institution's experience with attempted flap salvage for infected and exposed LVADs. Methods: A retrospective review for all LVAD-related infections treated with flaps at our institution from 2010 to 2015. Results: Twenty flaps were performed in 15 patients during the study period. Average age was 54 years. There were 4 women and 11 men with average body mass index of 30.6. Surgery was indicated for LVAD motor or drive line exposure in the setting of infection in all cases. Rectus abdominus (n = 10), omentum (n = 6), pectoralis major (n = 3), and intercostal (n = 1) were used for coverage. Complications resulted in approximately 67% of cases including hematoma (n = 4), seroma (n = 3), cellulitis (n = 1), and total flap loss (n = 1). Conclusions: Left ventricular assist devices are lifesaving interventions for patients with severe cardiac disease but are associated with a high rate of infectious complications over time. Although device coverage carries a high rate of complications, no devices required exchange due to infection or failed attempts at salvage.

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Breast Augmentation Combining Fat Injection and Breast Implants in Patients With Atrophied Breasts

imageAbstract: Breast augmentation is a challenging issue in atrophied breasts related to breastfeeding. Here, we describe a modification combining breast implants and fat grafting to obtain a successful outcome without implant edge visibility and palpability. Thirty-four women with atrophied breasts underwent breast augmentation over a 6-year period. The breast implants were placed in the subglandular plane to obtain optimal filling and expansion of atrophied breast tissue. Autologous fat grafting was performed above the inframammary sulcus to prevent implant visibility and palpability due to insufficient soft tissue coverage. Rippling, implant visibility, palpability, and capsular contracture were examined in the follow-up controls. Twenty anatomical and 48 round-shaped breast implants were placed with an average size of 321 cc (range, 200–415 cc). The mean injected fat volume was 114 cc (range, 98–142 cc) per breast. The mean follow-up period was 22 months (range, 19–31 months). The implants were palpable in only 1 breast; however, there was no implant visibility or rippling. Of the 68 breasts, 2 presented with cyst formations, and 4 with Becker Grade 2 capsular contracture, but no fat necrosis was observed. Overall, patient satisfaction in terms of breast shape, size, and coverage of the breast implant was 90%. A breast implant placed in the subglandular plane with fat injection throughout the lower quadrants of the breast may provide the desired increase in size and expansion of atrophied tissue while preventing implant visibility and palpability.

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Histone Deacetylase Inhibitors Enhance Cytotoxicity Towards Breast Tumors While Preserving the Wound-Healing Function of Adipose-Derived Stem Cells

imageIntroduction: Paclitaxel improves the oncologic response of breast cancer resections; however, it may negatively affect the wound-healing potential of human adipose-derived stem cells (hASCs) for fat grafting and reconstructive surgery. Histone deacetylase inhibitors (HDACis) modify the epigenetic regulation of gene expression and stabilize microtubules similarly to paclitaxel, thus, creating a synergistic mechanism of cell cycle arrest. We aim to combine these drugs to enhance cytotoxicity towards breast cancer cells, while preserving the wound-healing function of hASCs for downstream reconstructive applications. Methods: Triple negative breast cancer cells (MBA-MB-231) and hASCs (institutional review board–approved clinical isolates) were treated with a standard therapeutic dose of paclitaxel (1.0 μM) or with low-dose paclitaxel (0.1 μM) combined with the HDACi suberoylanilide hydroxamic acid or trichostatin A. Cell viability, gene expression, apoptosis, and wound-healing/migration were measured via methylthiazol tetrazolium assay, quantitative real-time polymerase chain reaction, annexin V assay, and fibroblast scratch assay, respectively. Results: Combined HDACi and low-dose paclitaxel therapy maintained cytotoxicity towards breast cancer cells and preserved adipose-derived stem cell viability. Histone deacetylase inhibitor demonstrated selective anti-inflammatory effects on adipose-derived stem cell gene expression and decreased expression of the proapoptotic gene FAS. Furthermore, HDACi therapy did not increase relative apoptosis within hASCs. A scratch assay demonstrated enhanced wound healing among injured fibroblasts indirectly co-cultured with HDACi-treated hASCs. Conclusions: Combining HDACi with low-dose paclitaxel improved cytotoxicity towards breast cancer cells and preserved hASC viability. Furthermore, enhanced wound healing was observed by improved migration in a fibroblast scratch assay. These results suggest that the addition of HDACi to taxane chemotherapy regimens may improve oncologic results and wound-healing outcomes after reconstructive surgery.

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Antimicrobial Prophylaxis Practice Patterns in Breast Augmentation: A National Survey of Current Practice

imageBackground: Breast augmentation is one of the most commonly performed aesthetic surgery procedures worldwide. Bacterial contamination is of paramount concern due to its role in biofilm formation and capsular contracture. Plastic surgeons have adopted a variety of perioperative interventions in efforts to reduce implant contamination, however definitive evidence supporting these practices is lacking. Objective: The goal of this study was to assess current practice patterns aimed at reducing implant-related infections in breast augmentation and comparing them to current evidence-based medicine recommendations. Methods: A 20-question survey assessing practices aimed at preventing breast implant associated infections was generated and distributed via SurveyMonkey to members of the American Society of Plastic Surgery whose electronic mail addresses were listed on the American Society of Plastic Surgery member website in April 2015. Results: A total of 253 responses were received from the survey. Of the respondents, 81% were in private practice. Respondents averaged 21 ± 9 years in practice, and 69 ± 111 breast augmentations performed per year. A majority of surgeons used Chlorhexidine to prepare the surgical site (45%) and triple antibiotic solution both as a soak for the implant before placement (40%) as well as for implant pocket irrigation (47%). A no-touch technique using the Keller Funnel was adopted by 42% of surgeons. Regarding antibiotic use in the postoperative period, first-generation cephalosporins were used by the majority of respondents (79%), and the most common duration was for 4 to 6 days (45%). Conclusions: Although there is heterogeneity in specific aspects of antimicrobial prophylaxis during breast augmentation, there is a clear trend toward practice modification geared towards preventing bacterial contamination in breast augmentation. Surgeons may benefit from high-level studies designed to create standardized evidence-based practice guidelines.

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