Impact of hydrodynamics on pollutant degradation and energy efficiency of VUV/UV and H2O2/UV oxidation processes.
The Vacuum-UV/UV process, an incipient catalyst/chemical-free advanced oxidation process (AOP), is potentially a cost-effective solution for the removal of harmful micropollutants from water. Utilizing a novel mechanistic numerical model, this work aimed to establish a thorough understanding of the degradation mechanisms in the VUV/UV process operating under continuous flow conditions, when compared with the widely applied H2O2/UV AOP. Of particular interest was the examination of the impact of flow characteristics (hydrodynamics) on the degradation efficacy of a target micropollutant during the VUV/UV and H2O2/UV AOPs. While hydroxyl radical (OH) oxidation was the dominant degradation pathway in both processes, the degradation efficacy of the VUV/UV process showed much stronger correlation with the extent of mixing in the photoreactor. Under a uniform flow regime, the degradation efficiency of the target pollutant achieved by the H2O2/UV process with 2- and 5 ppm H2O2 was greater than that provided by the VUV/UV process. Nonetheless, introduction of mixing and circulation zones to the VUV/UV reactor resulted in superior performance compared with the H2O2/UV AOP. Based on the electrical energy-per-order (EEO) analysis, incorporation of circulation zones resulted in a reduction of up to 50% in the overall energy cost of the VUV/UV AOP, while the corresponding reduction for the 5-ppm H2O2/UV system was less than 5%. Furthermore, the extent of OH scavenging of natural organic matter (NOM) on energy efficiency of the VUV/UV and H2O2/UV AOPs under continuous flow conditions was assessed using the EEO analysis.


Co-overexpression of native phospholipid-biosynthetic genes plsX and plsC enhances lipid production in Synechocystis sp. PCC 6803.
The overexpression of native plsX and plsC genes involving in fatty acid/phospholipid synthesis first timely-reported the significantly enhanced lipid contents in Synechocystis sp. PCC 6803. Growth rate, intracellular pigment contents including chlorophyll a and carotenoids, and oxygen evolution rate of all overexpressing (OX) strains were normally similar as wild type. For fatty acid compositions, saturated fatty acid, in particular palmitic acid (16:0) was dominantly increased in OX strains whereas slight increases of unsaturated fatty acids were observed, specifically linoleic acid (18:2) and alpha-linolenic acid (18:3). The plsC/plsX-overexpressing (OX + XC) strain produced high lipid content of about 24.3%w/dcw under normal condition and was further enhanced up to 39.1%w/dcw by acetate induction. This OX + XC engineered strain was capable of decreasing phaA transcript level which related to poly-3-hydroxybutyrate (PHB) synthesis under acetate treatment. Moreover, the expression level of gene transcripts revealed that the plsX- and plsC/plsX-overexpression strains had also increased accA transcript amounts which involved in the irreversible carboxylation of acetyl-CoA to malonyl-CoA. Altogether, these overexpressing strains significantly augmented higher lipid contents when compared to wild type by partly overcoming the limitation of lipid production.


Fabrication of a compound infrared microlens array with ultrashort focal length using femtosecond laser-assisted wet etching and dual-beam pulsed laser deposition.
We present the design, fabrication, and characterization of a compound infrared microlens array having an ultrashort focal length and a hyperbolic profile that comprises a PDMS microlens array and a GRIN lens. A concave microlens mold was first fabricated on a fused silica substrate using femtosecond laser irradiation followed by a wet etching process, and a standard replication process was employed to fabricate a PDMS convex lens array. To shorten the focal length further and cut off the visible spectrum of light, a graded DLC/silicon coating, which functioned as a GRIN lens and a visible light cut-off filter, was additionally deposited using dual-beam pulsed laser deposition. The lenslet diameter was 6 µm and the graded coating reduced the focal length from 4.5 to 2.9 µm.


Selection of a phylogenetically informative region of the norovirus genome for outbreak linkage.
The recognition of a common source norovirus outbreak is supported by finding identical norovirus sequences in patients. Norovirus sequencing has been established in many (national) public health laboratories and academic centers, but often partial and different genome sequences are used. Therefore, agreement on a target sequence of sufficient diversity to resolve links between outbreaks is crucial. Although harmonization of laboratory methods is one of the keystone activities of networks that have the aim to identify common source norovirus outbreaks, this has proven difficult to accomplish, particularly in the international context. Here, we aimed at providing a method enabling identification of the genomic region informative of a common source norovirus outbreak by bio-informatic tools. The data set of 502 unique full length capsid gene sequences available from the public domain, combined with epidemiological data including linkage information was used to build over 3,000 maximum likelihood (ML) trees for different sequence lengths and regions. All ML trees were evaluated for robustness and specificity of clustering of known linked norovirus outbreaks against the background diversity of strains. Great differences were seen in the robustness of commonly used PCR targets for cluster detection. The capsid gene region spanning nucleotides 900-1,400 was identified as the region optimally substituting for the full length capsid region. Reliability of this approach depends on the quality of the background data set, and we recommend periodic reassessment of this growing data set. The approach may be applicable to multiple sequence-based data sets of other pathogens.


Design of a single-chain polypeptide tetrahedron assembled from coiled-coil segments.
Protein structures evolved through a complex interplay of cooperative interactions, and it is still very challenging to design new protein folds de novo. Here we present a strategy to design self-assembling polypeptide nanostructured polyhedra based on modularization using orthogonal dimerizing segments. We designed and experimentally demonstrated the formation of the tetrahedron that self-assembles from a single polypeptide chain comprising 12 concatenated coiled coil-forming segments separated by flexible peptide hinges. The path of the polypeptide chain is guided by a defined order of segments that traverse each of the six edges of the tetrahedron exactly twice, forming coiled-coil dimers with their corresponding partners. The coincidence of the polypeptide termini in the same vertex is demonstrated by reconstituting a split fluorescent protein in the polypeptide with the correct tetrahedral topology. Polypeptides with a deleted or scrambled segment order fail to self-assemble correctly. This design platform provides a foundation for constructing new topological polypeptide folds based on the set of orthogonal interacting polypeptide segments.


Microcirculatory alterations: potential mechanisms and implications for therapy.
Multiple experimental and human trials have shown that microcirculatory alterations are frequent in sepsis. In this review, we discuss the characteristics of these alterations, the various mechanisms potentially involved, and the implications for therapy. Sepsis-induced microvascular alterations are characterized by a decrease in capillary density with an increased number of stopped-flow and intermittent-flow capillaries, in close vicinity to well-perfused capillaries. Accordingly, the surface available for exchange is decreased but also is highly heterogeneous. Multiple mechanisms may contribute to these alterations, including endothelial dysfunction, impaired inter-cell communication, altered glycocalyx, adhesion and rolling of white blood cells and platelets, and altered red blood cell deformability. Given the heterogeneous nature of these alterations and the mechanisms potentially involved, classical hemodynamic interventions, such as fluids, red blood cell transfusions, vasopressors, and inotropic agents, have only a limited impact, and the microcirculatory changes often persist after resuscitation. Nevertheless, fluids seem to improve the microcirculation in the early phase of sepsis and dobutamine also can improve the microcirculation, although the magnitude of this effect varies considerably among patients. Finally, maintaining a sufficient perfusion pressure seems to positively influence the microcirculation; however, which mean arterial pressure levels should be targeted remains controversial. Some trials using vasodilating agents, especially nitroglycerin, showed promising initial results but they were challenged in other trials, so it is difficult to recommend the use of these agents in current practice. Other agents can markedly improve the microcirculation, including activated protein C and antithrombin, vitamin C, or steroids. In conclusion, microcirculatory alterations may play an important role in the development of sepsis-related organ dysfunction. At this stage, therapies to target microcirculation specifically are still being investigated.


Cyclic nucleotides in the development of alcohol tolerance and dependence: a commentary.
This paper aims to outline some of the caveats inherent in studies on first and second messengers in the central nervous system as related to tolerance and dependence. Analysis of systems based on measures of first and second messengers may be an over-simplification, since the actual molecular consequences of neurotransmission might result in a multilevel cascade of many sequential messages including protein kinase activation and protein phosphorylation and/or dephosphorylation.


Comparative phylogenomics and phylotranscriptomics provide insights into the genetic complexity of nitrogen-fixing root-nodule symbiosis.
Plant root-nodule symbiosis (RNS) with mutualistic nitrogen-fixing bacteria is restricted to a single clade of angiosperms, the Nitrogen-Fixing Nodulation Clade (NFNC), and is best understood in the legume family. Nodulating species share many commonalities, explained either by divergence from a common ancestor over 100 million years ago or by convergence following independent origins over that same time period. Regardless, comparative analyses of diverse nodulation syndromes can provide insights into constraints on nodulation-what must be acquired or cannot be lost for a functional symbiosis-and the latitude for variation in the symbiosis. However, much remains to be learned about nodulation, especially outside of legumes. Here, we employed a large-scale phylogenomic analysis across 88 species, complemented by 151 RNA-seq libraries, to elucidate the evolution of RNS. Our phylogenomic analyses further emphasize the uniqueness of the transcription factor NIN as a master regulator of nodulation and identify key mutations that affect its function across the NFNC. Comparative transcriptomic assessment revealed nodule-specific upregulated genes across diverse nodulating plants, while also identifying nodule-specific and nitrogen-response genes. Approximately 70% of symbiosis-related genes are highly conserved in the four representative species, whereas defense-related and host-range restriction genes tend to be lineage specific. Our study also identified over 900 000 conserved non-coding elements (CNEs), over 300 000 of which are unique to sampled NFNC species. NFNC-specific CNEs are enriched with the active H3K9ac mark and are correlated with accessible chromatin regions, thus representing a pool of candidate regulatory elements for genes involved in RNS. Collectively, our results provide novel insights into the evolution of nodulation and lay a foundation for engineering of RNS traits in agriculturally important crops.


Validity and repeatability of anterior chamber depth measurements with Pentacam and Orbscan.
The purpose of this study was to determine the validity and repeatability of anterior chamber depth (ACD) measurements obtained with a novel rotating Scheimpflug camera (Pentacam; Oculus Optikgeräte GmbH, Wetzlar, Germany) to scanning slit topography (Orbscan; Bausch & Lomb, Rochester, NY). ACD in 60 healthy eyes was measured twice by two independent observers with each modality in random order. A total of eight measurements was performed on each eye. The mean differences between measurements, between observers, and between modalities were calculated, and 95% limits of agreement (LoA) were given as mean +/- 1.96 * standard deviation (SD) of the mean. The mean ACD values as determined with the different modalities (+/- SD) were 3.18 +/- 0.38 mm for Pentacam imaging and 3.23 +/- 0.40 for Orbscan. The mean difference between the two modalities was 0.047 mm (LoA: 0.176 to -0.081 mm). Within each modality, the limits of agreement of the differences between individual measurements and between different observers were within 0.073 mm or 4% of total ACD. The total variance in the observed data was 0.145. The variance resulting from interindividual differences in ACD was 85%, as a result of the two different modalities 15%, as a result of different observers 0.007%, and as a result of different measurements 0.007%. In the assessment of normal eyes, the differences of ACD values measured with Orbscan and Pentacam were within clinically acceptable levels, and inter- and intraobserver variability was considerably below clinically significant levels. Thus, these two modalities can be regarded as interchangeable.


Measuring carbon in forests: current status and future challenges.
To accurately and precisely measure the carbon in forests is gaining global attention as countries seek to comply with agreements under the UN Framework Convention on Climate Change. Established methods for measuring carbon in forests exist, and are best based on permanent sample plots laid out in a statistically sound design. Measurements on trees in these plots can be readily converted to aboveground biomass using either biomass expansion factors or allometric regression equations. A compilation of existing root biomass data for upland forests of the world generated a significant regression equation that can be used to predict root biomass based on aboveground biomass only. Methods for measuring coarse dead wood have been tested in many forest types, but the methods could be improved if a non-destructive tool for measuring the density of dead wood was developed. Future measurements of carbon storage in forests may rely more on remote sensing data, and new remote data collection technologies are in development.


Volume changes in the gallbladder in association with various radiologic examinations.
Our aim was to evaluate with sonography the volume changes of the gallbladder after intravenous administration of nonionic, iodinated contrast agents with or without ingestion of contrast agent or water. A total of 191 patients received intravenous injection of 50 ml of nonionic contrast agent (Group A), intravenous injection of 100 ml of contrast medium (Group B), intravenous injection of 100 ml of contrast medium together with ingestion of 1000 ml of a dilute solution of contrast medium (Group C), or 1000 ml of water taken orally (Group D). The gallbladder volume was measured by ultrasonography in the fasting state in all groups, 15 minutes after intravenous injection of contrast medium in Groups A, B, and C and 30 minutes after water ingestion in Group D. In all groups, the volume of the gallbladder was statistically lower after the procedure than before. There was no significant difference among the groups in terms of this gallbladder volume change. Intravenous injection of nonionic, iodinated contrast medium or ingestion of water or a dilute solution of contrast medium for various radiological examinations results in contraction of the gallbladder. Contrast-enhanced radiological examinations may interfere with sonographic evaluation of the gallbladder. If necessary, gallbladder ultrasonographic examination should be repeated with the patient in a fasting state. (c) 2005 Wiley Periodicals, Inc. J Clin Ultrasound 33:214-217, 2005.


Adrenocortical tumors in childhood: a report of four cases.
In the last 10 years, four children with adrenocortical tumors were treated in our hospital. Three of them had symptoms of adrenogenital virilizing syndrome, and another, nonfunctional, was found during a routine examination. A diagnosis of adrenocortical tumor was established given the symptoms, hormonal tests, and radiologic studies. Two of the tumors were located in the left side, and the histologic diagnosis was pleomorphic cortical adenoma; the ones that affected the right side were adenocarcinomas. All the patients were treated by surgery and none received chemotherapy. At both presurgical and postsurgical stages, the patients were treated with cortisol. All four children have had a favorable course, with normal growth and the disappearance of public hair and hirsutism; however, macrogenitals still persist.


Rapid determination of tetracycline antibiotics in serum by reversed-phase high-performance liquid chromatography with fluorescence detection.
A rapid and accurate determination of tetracycline antibiotics in human serum by reversed-phase high-performance liquid chromatography with fluorescence detection has been developed, based on protein precipitation in serum. Various reagents for precipitation were investigated, and 24% trichloroacetic acid in methanolic solution gave the maximum recovery (at least 94.3%) and interference-free chromatograms of different three tetracyclines. At a concentration of 0.5 micrograms/ml, the precision (relative standard deviation) ranged from 1.12 to 1.94%. In the range 0.04-10.0 micrograms/ml for oxytetracycline and chlorotetracycline and 0.01-10.0 micrograms/ml for tetracycline, linear responses were observed. The detection limits of this method were 10-35 ng/ml for all three antibiotics. The proposed method was applied to the determination of serum concentrations in subjects receiving tetracycline antibiotics.


Detection of SARS-CoV-2 and Other Viruses in Wastewater: Optimization and Automation of an Aluminum Hydroxide Adsorption-Precipitation Method for Virus Concentration.
This study aimed to provide a low-cost technique for virus detection in wastewater by improving an aluminum hydroxide adsorption-precipitation method. The releasing efficiency of viruses trapped by the aluminum hydroxide precipitates was improved by adding ethylenediaminetetraacetic acid disodium salt (EDTA-2Na) to dissolve the precipitates at a Na2EDTA·2H2O:AlCl3 molar ratio of 1.8-3.6. The recovery rates of the improved method for seven viruses, including SARS-CoV-2-abEN pseudovirus and six animal viruses, were 5.9-22.3% in tap water and 4.9-35.1% in wastewater. Rotavirus A (9.0-4.5 × 103 copies/mL), porcine circovirus type 2 (5.8-6.4 × 105 copies/mL), and porcine parvovirus (5.6-2.7 × 104 copies/mL) were detected in China's pig farm wastewater, while rotavirus A (2.0 × 103 copies/mL) was detected in hospital wastewater. SARS-CoV-2 was detected in hospital wastewater (8.4 × 102 to 1.4 × 104 copies/mL), sewage (6.4 × 10 to 2.3 × 103 copies/mL), and river water (6.6 × 10 to 9.3 × 10 copies/mL) in Nepal. The method was automized, with a rate of recovery of 4.8 ± 1.4% at a virus concentration of 102 copies/mL. Thus, the established method could be used for wastewater-based epidemiology with sufficient sensitivity in coping with the COVID-19 epidemic and other virus epidemics.


Identification of functional elements and reconstitution of the alpha 1(VI) collagen promoter.
To gain insight into the regulatory mechanisms of collagen VI synthesis we have characterized the cis-acting elements of the chicken alpha 1(VI) collagen promoter. Footprinting experiments with nuclear extracts from chicken embryos revealed three distinct elements, designated A, B, and C, that were protected from DNase I digestion. The nuclear proteins that interact with the three sites were identified by gel retardation assays in combination with the use of various oligonucleotide competitors as well as specific antibodies raised against well characterized transcription factors. Site A was found to be a target for transcriptional activator AP1, whereas sites B and C were shown to be recognized each by two distinct nuclear proteins which belong to the Sp1 multigene family. To address the question whether the three sites alone are able to direct transcription, a minipromoter construct was created in which the sequences of sites A, B, and C were placed in front of a reporter gene. After transfection into chicken fibroblasts, this construct exhibited a high relative promoter activity when compared to a large genomic fragment containing the basic alpha 1(VI) collagen promoter. Thus, the three sites are sufficient to induce transcription of this gene.


Investigation of the conformational lability of serum albumin macromolecules in aqueous solution by the NMR spin-echo method.
The complex proton spin-echo decay curve was recorded for human serum albumin (SA) solutions with different concentrations in normal and heavy water. The curve included three fast-decaying components for SA, in addition to the slow-decaying component for the water. The total amplitude of these three components roughly corresponded to the number of protons in the SA (with isotopic exchange taken into account); the component ratio remained constant at different concentrations and different temperatures between 4 and 39 degrees. The relatively slow-decaying protein component, which accounted for similar to 10% of the SA protons, was produced by the side chains of the protein. The presence of two other faster-decaying SA components with approximately equal amplitudes indicated that only about half of the remaining protons in the SA macromolecule are incorporated into the comparatively rigid globule, the other half belonging to groups with high conformational lability in aqueous solution. The activation energy for the aqueous component was close to that for pure water, while the activation energies for the protein components were roughly twice as large.


Upper extremity reconstruction by vascularized bone transfers: results and complications.
Twenty-one cases of vascularized fibula transfer for upper limb reconstruction are reviewed. Recipient sites included the humerus in 11 patients, radius in five, ulna in three, and clavicle in two. Overall, primary bone union occurred in 15 patients (71.4%), and eventual union was achieved in 17 patients (81%). Two of the patients in whom there was failure to unite are minimally symptomatic, with stable fibrous nonunions. The most favorable results were achieved in cases of forearm bone reconstruction (100% union) and for long-bone reconstruction after tumor resection (83% union).


Complications of retained epicardial pacing wires: an unusual bronchial foreign body.
The use of temporary epicardial pacing wires after open cardiac operations has been associated with a low morbidity. Complications can be divided into those encountered during placement, removal, or retention of these wires. We describe herein an unusual complication of a retained temporary epicardial pacing wire presenting as a posterior mediastinal mass and as a bronchial foreign body.


Functional deficiency of aryl hydrocarbon receptor augments oxygen toxicity-induced alveolar simplification in newborn mice.
Hyperoxia contributes to the development of bronchopulmonary dysplasia (BPD) in premature infants. New BPD is characterized as having alveolar simplification. We reported previously that aryl hydrocarbon receptor (AhR) deficiency increased susceptibility to hyperoxic lung injury in adult mice, and this was associated with decreased expression of cytochrome P450 1A enzymes and increased lung inflammation. Whether AhR protects newborn mice against hyperoxia-induced alveolar simplification is unknown. Thus, we tested the hypothesis that decreased activation of the pulmonary AhR augments hyperoxia-induced alveolar simplification and lung inflammation in newborn mice. Experimental groups included one-day old wild type (WT) and AhR dysfunctional (AhRd) mice exposed to 21% O₂ (air) or 85% O₂ (hyperoxia) for 14 days. Exposure of newborn WT mice to hyperoxia resulted in increased protein, enzyme and mRNA expression of the AhR-regulated lung cytochrome P450 1A1, NAD(P)H quinone oxidoreductase-1, and microsomal glutathione S-transferase 1 enzymes, suggesting that hyperoxia increases activation of the pulmonary AhR. On the other hand, in the AhRd mice, hyperoxia induced the AhR-regulated enzymes to a lesser extent probably due to the dysfunctional AhR in these mice. Alveolar simplification and lung inflammation was increased in mice exposed to hyperoxia compared with those exposed to air, and AhRd mice were more susceptible to hyperoxia-induced alveolar simplification and lung inflammation compared with WT mice. These findings suggest that decreased activation of the pulmonary AhR in newborn AhRd mice augments hyperoxia-induced alveolar simplification and lung inflammation in these mice.


Novel Quinazoline Derivatives Bearing Various 4-Aniline Moieties as Potent EGFR Inhibitors with Enhanced Activity Against NSCLC Cell Lines.
A class of novel quinazoline derivatives bearing various C-4 aniline moieties was synthesized and biologically evaluated as potent epidermal growth factor receptor (EGFR) inhibitors for intervention of non-small-cell lung cancer (NSCLC). Most of these inhibitors are comparable to gefitinib in inhibiting these cancer cell lines, and several of them even displayed superior inhibitory activity. In particular, analogue 5b with an IC50 of 0.10 μm against the EGFR wild-type A431 cells and 5c with an IC50 of 0.001 μm against the gefitinib-sensitive HCC827 cells (EGFR del E746-A750) was identified as highly active EGFR inhibitors. It was also significant that the discovered analogue 2f, not only has high potency against the gefitinib-sensitive cells (IC50 = 0.031 μm), but also possesses remarkably improved activity against the gefitinib-resistant cells. In addition, the enzymatic assays and the Western blot analysis for evaluating the effects of the typical inhibitors indicated that these molecules strongly interfere with the EGFR target.


Metabolic syndrome in relation to structure and function of large arteries: a predominant effect of blood pressure. A report from the SU.VI.MAX. Vascular Study.
Metabolic syndrome (MetS) is associated with increased risk of cardiovascular disease. However, the contribution of MetS to intermediate cardiovascular endpoints such as structure and function alterations of large arteries is still unclear. A total of 917 middle-aged French men and women participating to the SUpplémentation en VItamines et Minéraux AntioXydants (SU.VI.MAX) Vascular study were included. Carotid-femoral pulse wave velocity was used to assess aortic stiffness. Carotid ultrasound examination included measurements (at sites free of plaques) of intima-media thickness (IMT) at the common carotid arteries (CCA) and assessment of atherosclerotic plaques in the extracranial carotid arteries. Prevalence of the MetS defined by the National Cholesterol Education Program was 8.7%. Compared with subjects without MetS, subjects with MetS had significant higher mean values of CCA-IMT (P = .02) and pulse wave velocity (P = .0001). We found that MetS was not significantly related to the presence of carotid plaques. Blood pressure was the only MetS component associated with all vascular parameters. Addition of other MetS components in the multivariate models contributed poorly to the explained variance of these parameters. The MetS is adversely associated with markers of early arterial dysfunction, such as CCA-IMT and arterial stiffness. Although the associations between MetS and these arterial parameters are related to insulin resistance, blood pressure was shown to be the most important MetS component in relation to structure and function of large arteries.


Historical control data for the interpretation of ecotoxicity data: are we missing a trick?
Wildlife can be exposed to chemicals in the environment from various anthropogenic sources. Ecotoxicity studies, undertaken to address the risks from potential exposure to chemicals, vary in their design e.g. duration of exposure, effect types and endpoints measured. Ecotoxicity studies measure biological responses to test item exposure. Responses can be highly variable, with limited opportunity for control of extrinsic sources of variability. It is critical to distinguish between treatment-related effects and background 'normal variability' when interpreting results. Historical control data (HCD) can be a valuable tool in contextualising results from single studies against previous studies performed under similar conditions. This paper discusses the case for better use of HCD in ecotoxicology assessments, illustrating with case studies the value and difficulties of using HCD in interpretation of results of standard and higher-tier study designs. HCD are routinely used in mammalian toxicology for human health assessments, but not directly in ecotoxicology. The possible reasons for this are discussed e.g., different data types, the potential to mask effects, and the lack of guidance. These concerns are real but not insurmountable and we would like to see organisations such as OECD, EFSA and USEPA develop guidance on the principles of HCD collection. Hopefully, this would lead to greater use of HCD and regulatory acceptance. We believe this is not only a scientifically valid approach but also an ethical issue that is in line with societally driven legal mandates to minimise the use of vertebrate testing in chemical regulatory decision making.


Screening, diversity and partial sequence comparison of vegetative insecticidal protein (vip3A) genes in the local isolates of Bacillus thuringiensis Berliner.
Characterization, direct sequencing of the PCR amplicon and phylogenetic relationship was done to discover a novel Vip protein genes of the Bt isolates, to improve the prospects for insect control, more Vip proteins should be sought out and researched to predict their insecticidal activity. Characterization was based on direct sequencing of PCR amplicon using primers specific to vip3A gene was presented here. 12 out of 18 isolates screened were positive for vip gene-specific primers. Homology search for the partial sequences using BLAST showed that 11 isolates had high similarity to vip3Aa gene and only one fragment with vip3Ae gene (25-100% at nucleotide and amino acid level). Phylogenetic analysis showed that the gene sequences were responsible for geographic separation for divergence within vip genes, consistent with the evaluation of distinct bacterial population. Despite the geographical distances, strains harbouring vip genes have originated from common ancestors may significantly contribute to control resistant insect pests. Some strains have evolved to be quite distinct and others remain as members of closely related groups. The reported method is a powerful tool to find novel Vip3A proteins from large-scale Bt strains which is effective in terms of time and cost. Further the Vip proteins produced by different strains of B. thuringiensis are unique in terms of the sequence divergence and hence may also differ in their insecticidal activities.


Robot-assisted pancreatoduodenectomy with the da Vinci Xi: can the costs of advanced technology be offset by clinical advantages? A case-matched cost analysis versus open approach.
Robot-assisted pancreatoduodenectomy (RPD) has shown some advantages over open pancreatoduodenectomy (OPD) but few studies have reported a cost analysis between the two techniques. We conducted a structured cost-analysis comparing pancreatoduodenectomy performed with the use of the da Vinci Xi, and the traditional open approach, and considering healthcare direct costs associated with the intervention and the short-term post-operative course. Twenty RPD and 194 OPD performed between January 2011 and December 2020 by the same operator at our high-volume multidisciplinary center for robot-assisted surgery and for pancreatic surgery, were retrospectively analyzed. Two comparable groups of 20 patients (Xi-RPD-group) and 40 patients (OPD-group) were obtained matching 1:2 the RPD-group with the OPD-group. Perioperative data and overall costs, including overall variable costs (OVCs) and fixed costs, were compared. No difference was reported in mean operative time: 428 min for Xi-RPD-group versus 404 min for OPD, p = 0.212. The median overall length of hospital stay was significantly lower in the Xi-RPD-group: 10 days versus 16 days, p = 0.001. In the Xi-RPD-group, consumable costs were significantly higher (€6149.2 versus €1267.4, p < 0.001), while hospital stay costs were significantly lower: €5231.6 versus €8180 (p = 0.001). No significant differences were found in terms of OVCs: €13,483.4 in Xi-RPD-group versus €11,879.8 in OPD-group (p = 0.076). Robot-assisted surgery is more expensive because of higher acquisition and maintenance costs. However, although RPD is associated to higher material costs, the advantages of the robotic system associated to lower hospital stay costs and the absence of difference in terms of personnel costs thanks to the similar operative time with respect to OPD, make the OVCs of the two techniques no longer different. Hence, the higher costs of advanced technology can be partially compensated by clinical advantages, particularly within a high-volume multidisciplinary center for both robot-assisted and pancreatic surgery. These preliminary data need confirmation by further studies.


Characterization of spermidine and spermine synthases in Lotus japonicus: induction and spatial organization of polyamine biosynthesis in nitrogen fixing nodules.
The biosynthesis of the polyamines spermidine (Spd) and spermine (Spm) from putrescine (Put) is catalysed by the consequent action of two aminopropyltransferases, spermidine synthase (SPDS EC: 220.127.116.11) and spermine synthase (SPMS EC: 126.96.36.199). Two cDNA clones coding for SPDS and SPMS homologues in the nitrogen-fixing nodules of the model legume Lotus japonicus were identified. Functionality of the encoded polypeptides was confirmed by their ability to complement spermidine and spermine deficiencies in yeast. The temporal and spatial expression pattern of the respective genes was correlated with the accumulation of total polyamines in symbiotic and non-symbiotic organs. Expression of both genes was maximal at early stages of nodule development, while at later stages the levels of both transcripts declined. Both genes were expressed in nodule inner cortical cells, vascular bundles, and central tissue. In contrast to gene expression, increasing amounts of Put, Spd, and Spm were found to accumulate during nodule development and after maturity. Interestingly, nodulated plants exhibited systemic changes in both LjSPDS and LjSPMS transcript levels and polyamine content in roots, stem and leaves, in comparison to uninoculated plants. These results give new insights into the neglected role of polyamines during nodule development and symbiotic nitrogen fixation (SNF).


Nickel oral bioavailability in contaminated soils using a mouse urinary excretion bioassay: Variation with bioaccessibility.
To assess the health risk of nickel (Ni) in contaminated soils, studies rarely evaluated Ni bioavailability in the gastrointestinal (GI) tract, limiting the accurate regulation of contaminated sites. Here, for 15 soil samples contaminated by Ni-electroplating, Ni oral relative bioavailability (RBA, relative to NiSO4) was measured using a mouse urinary excretion bioassay. Nickel-RBA varied from 7.89% to 33.8% at an average of 19.1 ± 18.6%. The variation was not explained well by variation in soil properties including Ni speciation and co-contamination of other metals, which showed weak correlation with Ni-BRA (R2 < 0.36). In comparison, the Ni-RBA variation was explained well by the variation of soil-Ni solubility in simulated human gastric or gastrointestinal fluids, i.e., Ni bioaccessibility. Determined using the gastric (GP) and intestinal phases (IP) of solubility bioaccessibility research consortium (SBRC), physiologically based extraction test methods (PBET), and unified BARGE method (UBM), Ni bioaccessibility explained 54-71% variation of the Ni-RBA, suggesting that Ni oral bioavailability was predominantly controlled by Ni solubility in the GI tract. The results highlight the suitability of using simple, fast, and cost-effective bioaccessbility assays to predict site-specific Ni oral bioavailability.


Degradation of simulated Direct Orange-S (DO-S) textile effluent using nonthermal atmospheric pressure plasma jet.
One of the major environmental issues of textile industries is the discharge of large quantities of textile effluents, which are source of contamination of water bodies on surface of earth and quality of groundwater. The effluents are toxic, non-biodegradable, carcinogenic and prodigious threats to human and aquatic creatures. Since textile effluents can be treated efficiently and effectively by various advanced oxidation processes (AOPs). Among the various AOPs, cold atmospheric pressure plasma is a promising method among many prominent techniques available to treat the effluents. In this paper, we report about the degradation of simulated effluent, namely Direct Orange-S (DO-S) aqueous solution, using nonthermal atmospheric pressure plasma jet. The plasma treatment of DO-S aqueous solution was carried out as a function of various operating parameters such as potential and treatment time. The change in properties of treated DO-S dye was investigated by means of various analytical techniques such as high-performance liquid chromatography, UV-visible (UV-Vis) spectroscopy and determination of total organic content (TOC). The reactive species present in the samples were identified using optical emission spectrometry (OES). OES results confirmed that the formation of reactive oxygen and nitrogen species during the plasma treatment in the liquid surface was responsible for dye oxidation and degradation. Degradation efficiency, as monitored by color removal efficiency, of 96% could be achieved after 1 h of treatment. Concurrently, the TOC values were found to decrease with plasma treatment, implying that the plasma treatment process enhanced the non-toxicity nature of DO-S aqueous solution. Toxicity of the untreated and plasma-treated dye solution samples was studied using Escherichia coli (E. coli) and Staphylococcus (S. aureus) organisms, which demonstrated that the plasma-treated dye solution was non-toxic in nature compared with untreated one.


Modelling of Brain Deformation After Decompressive Craniectomy.
Hyperelastic finite element models, with either an idealized cylindrical geometry or with realistic craniectomy geometries, were used to explore clinical issues relating to decompressive craniectomy. The potential damage in the brain tissue was estimated by calculating the volume of material exceeding a critical shear strain. Results from the idealized model showed how the potentially damaged volume of brain tissue increased with an increasing volume of brain tissue herniating from the skull cavity and with a reduction in craniectomy area. For a given herniated volume, there was a critical craniectomy diameter where the volume exceeding a critical shear strain fell to zero. The effects of details at the craniectomy edge, specifically a fillet radius and a chamfer on the bone margin, were found to be relatively slight, assuming that the dura is retained to provide effective protection. The location in the brain associated with volume expansion and details of the material modeling were found to have a relatively modest effect on the predicted damage volume. The volume of highly sheared material in the realistic models of the craniectomy varied roughly in line with differences in the craniectomy area.


Jubanines F-J, cyclopeptide alkaloids from the roots of Ziziphus jujuba.
Five Ib-type cyclopeptide alkaloids, jubanines F-J (1-5), and three known compounds, nummularine B (6), daechuine-S3 (7), and mucronine K (8) were isolated from the roots of Ziziphus jujuba. Their structures were fully characterized by spectroscopic analyses in combination with chemical derivatization. Compounds 1-3, and 6 were evaluated for their antiviral activity against the porcine epidemic diarrhea virus (PEDV). Compounds 2, 3, and 6 showed potent inhibitory effects on PEDV replication.


Segmenting functional tissue units across human organs using community-driven development of generalizable machine learning algorithms.
The development of a reference atlas of the healthy human body requires automated image segmentation of major anatomical structures across multiple organs based on spatial bioimages generated from various sources with differences in sample preparation. We present the setup and results of the Hacking the Human Body machine learning algorithm development competition hosted by the Human Biomolecular Atlas (HuBMAP) and the Human Protein Atlas (HPA) teams on the Kaggle platform. We create a dataset containing 880 histology images with 12,901 segmented structures, engaging 1175 teams from 78 countries in community-driven, open-science development of machine learning models. Tissue variations in the dataset pose a major challenge to the teams which they overcome by using color normalization techniques and combining vision transformers with convolutional models. The best model will be productized in the HuBMAP portal to process tissue image datasets at scale in support of Human Reference Atlas construction.


Curcumin nanocrystals attenuate cyclophosphamide-induced testicular toxicity in mice.
The cancer therapy using cyclophosphamide (CP) has been associated with adverse effects on the testicular function that raises concerns about the future fertility potential among cancer survivors. Curcumin, a polyphenol, has shown to possess a plethora of biological functions including tissue protective effects. In the present study, we investigated the protective effects of curcumin nanocrystals (NC) in mitigation of CP-induced testicular toxicity. Healthy adult (8-10 week) and prepubertal (2 week) male Swiss albino mice were injected with a single dose of CP (200 mg/kg) intraperitoneally (i.p). NC (4 mg/kg, i.p.) was administered every alternate day, for 35 days in adult mice while, a single dose of NC was injected intraperitoneally to prepubertal mice 1 h prior to CP. Administration of multiple doses of NC ameliorated CP-induced testicular toxicity in adult mice, which was evident from the improved sperm functional competence, sperm chromatin condensation, seminiferous tubule architecture and decreased apoptosis in testicular cells. Further, administration of NC 1 h prior to CP in prepubertal mice modulated the expression of genes pertaining to proliferation, pluripotency, DNA damage and DNA repair in spermatogonial cells at 24 h after the treatment. Overall, these results suggest that NC could be a promising chemoprotective agent, which can have potential application in male fertility preservation.


Molecular sensing by pyrene excimer signal of fluorescent DNA aptamer.
The pyrene excimer label (2) has been incorporated into the adjacent residue to the binding site of anti-ATP DNA aptamer. The fluorescent aptamer exhibited the specific fluorescence signal only in the presence of ATP.


Polymerization force-regulated actin filament-Arp2/3 complex interaction dominates self-adaptive cell migrations.
Cells migrate by adapting their leading-edge behaviors to heterogeneous extracellular microenvironments (ECMs) during cancer invasions and immune responses. Yet it remains poorly understood how such complicated dynamic behaviors emerge from millisecond-scale assembling activities of protein molecules, which are hard to probe experimentally. To address this gap, we establish a spatiotemporal "resistance-adaptive propulsion" theory based on the interactions between Arp2/3 complexes and polymerizing actin filaments and a multiscale dynamic modeling system spanning from molecular proteins to the cell. We quantitatively find that cells can accurately self-adapt propulsive forces to overcome heterogeneous ECMs via a resistance-triggered positive feedback mechanism, dominated by polymerization-induced actin filament bending and the bending-regulated actin-Arp2/3 binding. However, for high resistance regions, resistance triggers a negative feedback, hindering branched filament assembly, which adapts cellular morphologies to circumnavigate the obstacles. Strikingly, the synergy of the two opposite feedbacks not only empowers the cell with both powerful and flexible migratory capabilities to deal with complex ECMs but also enables efficient utilization of intracellular proteins by the cell. In addition, we identify that the nature of cell migration velocity depending on ECM history stems from the inherent temporal hysteresis of cytoskeleton remodeling. We also show that directional cell migration is dictated by the competition between the local stiffness of ECMs and the local polymerizing rate of actin network caused by chemotactic cues. Our results reveal that it is the polymerization force-regulated actin filament-Arp2/3 complex binding interaction that dominates self-adaptive cell migrations in complex ECMs, and we provide a predictive theory and a spatiotemporal multiscale modeling system at the protein level.


Tachykinins amplify the action of capsaicin on central histaminergic neurons.
Substance P (SP), a product of the tachykinin 1 (Tac1) gene, is expressed in many hypothalamic neurons. Its wake-promoting potential could be mediated through histaminergic (HA) neurons of the tuberomamillary nucleus (TMN), where functional expression of neurokinin receptors (NKRs) waits to be characterized. As in the process of nociception in the peripheral nervous system (PNS) capsaicin-receptor (transient potential vanilloid 1: TRPV1) signalling is amplified by local release of histamine and SP, we tested the involvement of tachykinins in the capsaicin-induced long-lasting enhancement (LLEcaps) of HA neurons firing by investigating selective neurokinin receptor ligands in the hypothalamic mouse brain slice preparation using patch-clamp recordings in cell-attached mode combined with single-cell RT-PCR. We report that the majority of HA neurons respond to SP (EC50 3 nM), express the SP precursor tachykinin 1 (Tac1) gene and at least one of the neurokinin receptors. Responses to selective agonists of three known neurokinin receptors were sensitive to corresponding antagonists. LLEcaps was significantly impaired by the neurokinin receptor antagonists, indicating that in hypothalamus, as in the PNS, release of tachykinins downstream to TRPV1 activation is able to boost the release of histamine. The excitatory action of SP on histaminergic neurons adds another pathway to the noradrenergic and orexinergic ones to synergistically enhance cortical arousal. We show NK1R to play a prominent role on HA neurons and thus the control of wakefulness.


Treatment of panic disorder by trigeminal nerve manipulation: A case series.
This is a report on two cases of patients with acute severe panic disorder relieved of their symptoms by manual manipulations of the trigeminal nerve's alveolar branches. The manipulations were performed via the oral cavity during one session, or two consecutive sessions less than a week apart. No other effective treatment was administered prior, concurrently or since the time of the treatment. The recovery from panic disorder was immediate and lasted for the entire period of observation of three years. The authors used the same procedure and achieved identical clinical results treating ten other clients over a period of three years. This was not a planned experiment or randomized study. Rather, this report presents clinical evidence and the authors' hypothesis based on clinical data and literature review.


Determinants of the repetitive-CMAP occurrence and therapy efficacy in slow-channel myasthenia.
To find determinants of the occurrence of repetitive compound muscle action potential (R-CMAP) and to assess the efficacy of channel blocker therapy in slow-channel congenital myasthenic syndrome (SCCMS). Neurologic examination, EMG study, laboratory test, muscle biopsy, and next-generation and Sanger sequencing; literature review of reported patients with SCCMS, including EMG, kinetics of mutant acetylcholine receptors (AChRs), and response to therapy; and simulation of the decay phase of endplate potential (EPP) were performed. Three newly characterized and 57 reported patients with SCCMS with mutations of AChR subunits were included. In patients with R-CMAP, the length of channel opening bursts of mutant AChR was increased 8.68 ± 2.82 (mean ± SD)-fold compared to wild-type; in patients without R-CMAP, the length was increased 3.84 ± 0.65-fold (95% confidence interval 3.18-6.50, p = 0.000014). The EPP amplitude after refractory period of action potential in muscle fiber is above the threshold in patients with R-CMAP but below the threshold in patients without R-CMAP. In patients with good results from channel blocker therapy, treatment was initiated 11.60 ± 5.17 years after onset of symptoms; in patients with no to moderate benefit from channel blocker therapy, treatment was initiated 30.70 ± 12.72 years after onset (95% confidence interval -28.57 to -9.63, p = 0.00089). In SCCMS, the R-CMAP occurrence is related to the extent of prolongation of the opening episodes of mutant AChR channel. Channel blocker treatment is more effective the sooner it is started after the onset of symptoms. This study provides Class IV evidence that channel blocker therapy in patients with SCCMS improves symptoms.


[ Role of meteorological factors in the formation of the plant nitrate pool].
Meteorological conditions during the period of plant vegetation make the greatest contribution to the formation of the plant nitrate pool. Nitrate nitrogen is accumulated at high concentrations during sunless, cold years. The weather conditions of the last five to ten days of plant vegetation determine nitrate accumulation. The meteorological conditions during the entire period of vegetation determine the increase in the content of nitrates due to nitrogen fertilizers. The effect of meteorological conditions is especially pronounced in low fertility soils and upon introduction of high doses of nitrogen fertilizers. The reaction of plants to unfavorable meteorological conditions greatly varies with reference to their biological and genotypical specificity.


Mechanism of human lymphotoxin and tumor necrosis factor induced destruction of cells in vitro: phospholipase activation and deacylation of specific-membrane phospholipids.
The role of phospholipase (PLase) activation and lipid metabolism in lymphotoxin (LT)- and tumor necrosis factor (TNF)-mediated destruction of murine L929 cells was examined. At the levels of LT and TNF employed, cell destruction began at 4-6 h and was 99% complete by 30 h. Cell membrane phospholipids (PL), labelled in situ at the C2 position with 14C arachidonic acid, were analyzed by two-dimensional thin-layer chromatography and quantitated over a 30 h time course after LT or TNF treatment. The ratio of radiolabel incorporation relative to the actual amount of each PL present was determined by inorganic phosphate analysis. Radiolabelled arachidonic acid, eicosanoids, and neutral lipids were released into the medium prior to the onset of cell death (4-6 h) and continued to accumulate linearly throughout the destructive reaction. There was a quantitative relationship between the appearance of radiolabelled metabolites in the media and the loss of radiolabelled cellular PL. Cellular phosphatidylethanolamine was the primary PL deacylated by PLase action, showing a 75% reduction in radiolabel. The PLase inhibitors--quinacrine, hydrocortisone, dexamethasone, and indomethacin--were potent inhibitors of LT- and TNF-mediated cell destruction, suggesting that selective deacylation of specific membrane PL by PLase activation is an important step in the events that lead to LT- and TNF-mediated cellular destruction in vitro.


Zero-valent iron enhanced in-situ advanced anaerobic digestion for the removal of antibiotics and antibiotic resistance genes in sewage sludge.
The in-situ advanced anaerobic digestion (AAD) enhanced with zero-valent iron powder (ZVI) under mesophilic condition was investigated to remove 5 antibiotics (sulfamerazine (SMR), sulfamethoxazole (SMZ), ofloxacin (OFL), tetracycline (TC), and roxithromycin (ROX)) and 11 antibiotic resistance genes (ARGs) (AAC (6')-IB-CR, qnrS, ermF, ermT, ermX, sul1, sul2, sul3, tetA, tetB, and tetG) in sewage sludge. The effects of different ZVI dosages, antibiotic concentrations, and solid retention time (SRTs) on the removal were explored. Also, the correlation coefficient of antibiotics and ARGs, microbial community structure, biogas production and methane yield were analyzed. All conducted treatments operated stably, and the modified Gompertz model described the cumulative methane yield well. The antibiotics, with the exception of OFL, were effectively removed in the sewage sludge at a dosage of 1000 mg/L ZVI, SRT 20 d, and an antibiotic concentration of 20 μg/L during AAD. The removal rates of SMZ, SMR, TC, and ROX reached 97.39%, 74.54%, 78.61%, and 56.58%, respectively. AAC (6')-IB-CR and tetB could be effectively reduced during the in-situ AAD. Through the redundancy analysis, AAC (6')-IB-CR, ermT, ermX, sul2, tetB, and tetG had strong positive correlations with the antibiotics in the reactor. The principle component analysis revealed that the community structure was similar when the SRT was 10 d and 20 d at the same amount of ZVI and antibiotic concentrations in the sludge. Under the operating parameters of 1000 mg/L ZVI dosage, SRT 20 d, and an antibiotic concentration of 20 μg/L, Erysipelotrichia, Verrucomicrobia, Clostridia, Caldiserica, and Alphaproteobacteria of the class were dominated microorganisms in the anaerobic digestion.


In vitro assembly of poliovirus empty capsids: antigenic consequences and immunological assay of the morphopoietic factor.
The assembly of poliovirus 14 S particles into empty capsids was studied without cell extract (self-assembly) and in extracts of infected or uninfected HeLa cells. The products were analyzed using monoclonal antibodies specific for N1, N2, or H epitopes. The empty capsids formed in infected cell extract, and only those, possessed N2 epitopes like the procapsids formed in vivo, thus showing that a virally encoded or induced factor determines the antigenicity of the assembly product. Based on this observation, a simple immunological assay for the activity of the morphopoietic factor is presented. This factor is shown to lack serotype specificity.


No association between hormonal abnormality and sexual dysfunction in Japanese schizophrenia patients treated with antipsychotics.
Although sexual dysfunction is believed to be caused by hormonal abnormalities, few reports have studied sexual dysfunction and its association with hormonal abnormalities in Asian populations with schizophrenia. We employed a cross-sectional, case-control survey design to collect data from 191 (108 men) Japanese schizophrenia outpatients treated with antipsychotics and 182 (49 men) healthy subjects. Sexual dysfunction was evaluated using the Udvalg for Kliniske Undersøgelser (UKU) Side Effect Rating Scale. We measured plasma concentrations of prolactin in both genders and testosterone in men and estradiol in women. Multiple regression analyses revealed the following findings: the number of antipsychotics correlated with diminished sexual desire (standardized beta = 0.241, p < 0.05); the dose of antipsychotics correlated with gynecomastia (standardized beta = 0.277, p < 0.01), increased sexual desire (standardized beta = 0.229, p < 0.05), and ejaculatory dysfunction (standardized beta = 0.248, p < 0.05); and the dose of antipsychotics correlated with menorrhagia in women (standardized beta = 0.284, p < 0.05). However, neither plasma concentrations of prolactin, testosterone nor estradiol correlated with sexual dysfunction. The present study demonstrated that an association between sex hormone abnormalities and sexual dysfunction is unlikely but that the dose or number of antipsychotics is associated with some sexual dysfunction.


Identification of P-selectin glycoprotein ligand-1 as a useful marker in acute myeloid leukaemias.
Immunophenotyping is considered to be less valuable in the diagnosis of acute myeloid leukaemias (AML) compared with acute lymphoid leukaemias. Here, we present data on the use of quantitative flow cytometry (QFC) of P-selectin glycoprotein ligand 1 (PSGL-1, CD162) and three-colour immunophenotyping including CD162 staining in the identification of myeloid precursors in AML. Analysis of normal peripheral blood (n = 20) and normal bone marrow (n = 5) samples and on 20 samples from de novo M1, M2, M4 and M5 AML patients demonstrated that PSGL-1 is differentially expressed on various mature and immature leucocyte subsets. It was found by QFC that neutrophils expressed 26500 +/- 4500 and monocytes 47200 +/- 9900 copies of PSGL-1 on their surface, whereas AML blasts from M1 and M2 AML patients expressed significantly less PSGL-1 (12 000 +/- 5300) than mature neutrophils (P < 0.001). In M4 and M5 leukaemias, however, the amount of PSGL-1 on monocytic precursors is displayed in a fairly broad range that is not significantly different from that of mature monocytes (P = 0.084). Using three-colour immunophenotyping PSGL-1-dim staining was co-expressed with CD7 and C34 positivity and PSGL-1 staining intensity on immature myeloid cells paralleled with CD45 expression. This would imply a differential expression of PSGL-1 during myeloid haematopoietic development and suggests that quantification of surface PSGL-1 may aid in differentiating myeloblasts from monoblasts by immunophenotyping in different AML subsets.


Genome-wide association study in Guillain-Barré syndrome.
Guillain-Barré syndrome (GBS) is considered to have an immune-mediated basis, but the genetic contribution to GBS is unclear. We conducted a GWAS involving 215 GBS patients and 1,105 healthy controls. No significant associations of individual SNPs or imputed HLA types were observed. We performed a genome-wide complex trait analysis for evaluation of the heritability of GBS, and found that common SNPs contribute up to 25% of susceptibility to the disease. Genetic risk score analysis showed no evidence of overlap in genetic susceptibility factors of GBS and multiple sclerosis. Given the unexplained heritability of the trait further larger GWAS are indicated.


Current Feeding Practice of Xenopus laevis in a Laboratory Setting.
African clawed frogs are common animal models used in various research areas. However, husbandry and especially feeding regimens are not nearly as standardized as is established for other laboratory animals. We recorded the diets and feeding protocols commonly used in laboratory practice in a questionnaire (18 responses). The survey revealed a wide variety of housing conditions. Feeding protocols and, in particular, diet composition varied considerably between facilities. While diets tailored to Xenopus were used in the majority, differences in feeding frequency and dietary components were noted. From five responses, the weekly feed intake per frog could be calculated, showing considerable differences in dry matter intake (1.37-5.4 g). The labelled nutrient content of the diets fed in the facilities (n = 10) met the recommendations in most cases, with protein as the major energy source. However, the mineral content varied markedly between diets. Both floating and sinking diets were used, while quickly sinking diets were associated with feed leftovers. Feed processing may likely influence feed intake behavior. Further research is needed to ensure standardization for aquatic species with respect to husbandry systems, feeding regimens, and especially the nutrient composition of feeds. Furthermore, this work will contribute positively to animal welfare and the comparability of research results.


Control of a one-link arm by burst signal generators.
The focus of this paper is the study of stability and point-to-point movement of a one-link arm. The sagittal arm has two musculotendon actuators, two neural oscillators that generate burst signals as motoneuron inputs, and spindles and Golgi tendon organs for extrinsic reflex feedbacks. It is shown that coactivation leads to intrinsic position and velocity feedback, and that the tendons introduce intrinsic force and rate of force feedback. In addition, the integrating effects of the tendons are studied when the actuator is constructed from a large number of identical fibers that are excited by alpha signals whose arrival times at the fiber are randomly distributed. Each of the musculotendon actuators receives two input signals--a burst signal analogous to alpha inputs and a conventional analogue signal that represents fusimotor input to the spindles. The process of combining burst signals and conventional analogue signals is studied. Simulation results show that the movement of the system with burst signals as inputs has overshoot and speed similar to the system with analogue signals.


Hand Therapy Treatment.
Clinical pain is multifactorial. It is not simply the consequences of a "switching on" of the pain system in the periphery, but also excitability of central nociceptive areas. For pain management to be successful in treating the upper extremity both the peripheral and central symptoms must be targeted. The patient education process must ensure that patients understand their symptoms and treatment program. This article discusses recent advancements in the neuroscience of pain that impact evolving strategies to identify and treat the pain mechanisms.


You're the Flight Surgeon.
Moore JL, Jackson CR, Ellis JC, Norrid C. You're the flight surgeon: myelolipoma. Aerosp Med Hum Perform. 2015; 86(4):421-423.


New antiangiogenetic agents and non-small cell lung cancer.
New blood vessel formation, known as angiogenesis is a fundamental event in the process of tumor growth and metastatic dissemination. Due to its central role in tumor angiogenesis, the vascular endothelial growth factor (VEGF) and its receptor have been a major focus of basic research and drug development in the field of oncology, including the treatment of non-small cell lung cancer (NSCLC). Approaches targeting VEGF include monoclonal antibodies and vascular endothelial growth factor receptor-tyrosine kinase inhibitors (VEGFR-TKIs). Bevacizumab (Avastin) is an anti-VEGF recombinant humanized monoclonal antibody. A very recent randomized phase III trial demonstrated a statistically significant advantage in median survival favouring the combination of bevacizumab plus chemotherapy versus chemotherapy alone in the treatment of advanced non-squamous NSCLC. This study represents the first evidence of superior efficacy of targeted therapy combined with chemotherapy over chemotherapy alone in the treatment of NSCLC. ZD6474 is an orally bioavailable inhibitor of VEGFR-2 tyrosine kinase. First evidences of antitumor activity and its excellent toxicity profile make it a promising targeted agent for the treatment of NSCLC. A recent phase I/II study examined the combination of Epidermal Growth Factor Receptor (EGFR)-TKI erlotinib and bevacizumab in patients with non-squamous stage IIIB/IV NSCLC. Data on antitumor activity of this combination have to be considered very promising. Clinical trials of multiple targeted therapy may represent the second generation studies in the treatment of NSCLC.


[Monoamine oxidase activity and ultrastructure of subcellular fractions of the cerebral cortex under normal conditions and in the presence of phenamine stimulation].
Crude mitochondrial fractions were fractionated into purified mitochondria, heavy- and light synaptosomes and synaptosomal membranes. The highest MAO activity was observed in purified mitochondria and heavy synaptosomes. Amphetamine strongly inhibited the MAO activity in heavy synaptosomes, and only slightly affected light synyptosomes. Electron microscopy of heavy synaptosomes revealed specific vesicles suggesting the presence of nerve endings of monoaminergic neurons, and the increase of the functional activity of these neurons.


The levels of cyclic GMP and glucose 1,6-diphosphate, and the activity of phosphofructokinase, in muscle from normal and dystrophic mice.
A striking reduction in the levels of glucose 1,6-diphosphate and an increase in cyclic GMP were found in muscle from dystrophic mice. Concomitant to these changes, the allosteric activity of phosphofructokinase was found to be markedly reduced. These findings could offer an explanation for the observed reduction in glycolysis in the dystrophic muscle.


Determination of trace elements (Cu, Zn, Mn, Pb) and magnesium by atomical absorption in patients receiving total parenteral nutrition.
We measured the serum levels of four trace elements (Cu, Zn, Mn, Pb) and Mg in surgical patients receiving total parenteral nutrition (TPN). The clinical implications and the results are discussed. Two groups of patients were studied: the first group (n = 40) was our study group and the second (n = 40) was the control group. Four measurements of each trace element (TE) in blood serum were carried out: one before initiating TPN, one 24 h after, one 3 d later, and the last one immediately after discontinuing TPN. Each measurement was repeated twice. The Perkin-Elmer atomic absorption spectrophotometer (model 2380) with furnace graphite HGA-300 was used to measure the TE levels and an acetylene flame was used to measure the Mg levels. Levels of all the TEs, except Pb, were lower before the administration of TPN compared with the control group (P < 0.05). The levels of TEs during and immediately after TPN were generally lower in comparison with the initial measurement before the administration of TPN. The results of this study suggest that it may be necessary to 1) add Cu, Zn, Mn, and Mg to the parenteral nutritional solution and 2) follow the fluctuations in serum levels during the administration of TPN.


Effect of 5'-methylthioadenosine on RNA and DNA synthesis in cultured Xenopus laevis cells.
We have examined the effect of 5'-methylthioadenosine (MTA) on RNA metabolism and DNA synthesis in a cultured Xenopus laevis epithelial cell line. Cells incubated in the presence of MTA were transcriptionally less active than controls as measured by the run-off transcription technique. Experiments which measured the percentage of newly-synthesized poly(A)+ RNA and alpha-amanitin-sensitive nuclear transcripts suggested that MTA inhibited transcription by both RNA polymerases I and II. DNA synthesis was observed to proceed normally at times when MTA inhibited RNA synthesis. These results suggest that the well-known anti-proliferative effect of MTA is due indirectly to its initial effects on the transcription of ribosomal RNA (rRNA) and messenger RNA (mRNA) rather than a direct suppression of DNA replication.


Intrathecal administration of 5-HT(3) receptor agonist modulates jaw muscle activity evoked by injection of mustard oil into the temporomandibular joint in the rat.
The effect of intrathecal administration of the 5-HT(3) receptor agonist 2-methyl-5-hydroxytryptamine (2m-5HT) on jaw muscle activity evoked by mustard oil (MO) injection into the temporomandibular joint of anesthetized rats was examined. One microgram or 100 microg of 2m-5HT significantly enhanced or suppressed jaw muscle responses, respectively. Pre-administration of tropisetron, a 5-HT(3) receptor antagonist, attenuated the effect of 2m-5HT. These results indicate that activation of 5-HT(3) receptors can modulate trigeminal nociceptive responses.


Immune response at 12-23 months following a single dose of Vero cell culture-derived Japanese encephalitis (JE) vaccine in adults previously vaccinated with mouse brain-derived JE vaccine.
Japanese encephalitis (JE) virus is an important cause of neurological disease in Asia. JE vaccine is recommended for travelers with higher JE risk itineraries. Inactivated Vero cell culture-derived JE vaccine (JE-VC) is the only JE vaccine currently available in the United States. An inactivated mouse brain-derived JE vaccine (JE-MB) previously was available but production was discontinued. One JE-VC dose administered to adults previously vaccinated with ≥3 doses of JE-MB provides good short-term protection for at least one month, but data on longer-term protection are limited. We evaluated non-inferiority of the JE virus neutralizing antibody response at 12-23 months in JE-MB-vaccinated adults administered one JE-VC dose compared with JE vaccine-naïve adults administered a JE-VC two-dose primary series. We obtained archived sera from U.S. military personnel and performed a 50% plaque reduction neutralization test for anti-JE virus neutralizing antibodies. We compared the geometric mean titer (GMT) and seroprotection rate at 12-23 months after one JE-VC dose in previously JE-MB-vaccinated personnel and after the second JE-VC dose in previously JE vaccine-naïve personnel. Non-inferiority was concluded if the lower bound of the two-sided 95% confidence interval (CI) of the GMT ratio in previously vaccinated to vaccine-naïve personnel was >1/1.5. The GMT in previously JE-MB-vaccinated persons was 75 (95% CI 63-90) and in previously JE vaccine-naïve persons was 12 (95% CI 11-14), and seroprotection rates were 94% (235/250) and 54% (135/250), respectively. The ratio of GMTs was 6.3 (95% CI: 5.0-7.7), satisfying the criterion for non-inferiority. One JE-VC dose in previously JE-MB-vaccinated military personnel provides good protection for at least 1-2 years. The benefits of administration of a single JE-VC dose in previously JE-MB-vaccinated adults include a shorter time to completion of re-vaccination before travel, a decrease in the risk of adverse events, and reduced costs.


The Role and Benefits of Dermocosmetics in Acne Management in Japan.
In Japan, as in other countries around the world, acne vulgaris is a common disease and a frequent reason for patients to consult dermatologists. For optimal management of acne, it is important to understand how available products to support skin health can be used both with and without prescription products. Dermocosmetics can be defined as skincare agents with dermatologically active ingredients that directly support or care for the symptoms of various skin conditions (distinct from vehicle effects). There are products with active ingredients-including familiar ones such as niacinamide, retinol derivatives, and salicylic acid-that target important aspects of acne pathophysiology. Others, including ceramides, glyercin, thermal spring water, and panthenols, may have positive effects on skin barrier function that are useful in managing acne. This publication will discuss the roles of dermocosmetics in acne either as monotherapy to manage the milder forms of acne and help prevent relapses, or as adjuncts to prescription therapy to increase efficacy or adherence and assist in prevention of local adverse effects. Dermocosmetics may also have active ingredients that positively impact the skin microbiome.


On the molecular interactions between fibrin, tissue-type plasminogen activator and plasminogen.
The molecular interactions involved in the fibrin-mediated stimulation of plasminogen activation by tissue-type plasminogen activator (t-PA) were studied using natural human plasminogen (nPlg) and rPlg-Ala740, a recombinant human plasminogen in which the catalytic site is destroyed by mutagenesis of the active site Ser740 to Ala. Using this rPlg-Ala740 moiety, the dissociation constant of the interaction between plasminogen and CNBr-digested fibrinogen was determined to be 0.40 microM. In addition, conversion of 125I-labeled single chain plasminogen to two chain plasmin by single chain recombinant t-PA (rt-PA) in the absence or the presence of CNBr-digested fibrinogen was quantitated on reduced SDS-gel electrophoresis, combined with autoradiography and radioisotope counting of gel bands. In the absence of fibrin, the activation rate of nPlg and rPlg-Ala740 by single-chain rt-PA was comparable. In the presence of fibrin, however, the activation rate of rPlg-Ala740 was about 20-fold lower than that of nPlg. These results with rPlg-Ala740 may be explained by an impaired formation of the stable cyclic ternary complex between plasminogen, t-PA and fibrin, which mediates the fibrin stimulation of plasminogen activation by t-PA or, alternatively, by impaired conversion of single chain rt-PA to two chain rt-PA at the fibrin surface.


The mouse nicotinamide mononucleotide adenylyltransferase chaperones diverse pathological amyloid client proteins.
Molecular chaperones safeguard cellular protein homeostasis and obviate proteotoxicity. In the process of aging, as chaperone networks decline, aberrant protein amyloid aggregation accumulates in a mechanism that underpins neurodegeneration, leading to pathologies such as Alzheimer's disease and Parkinson's disease. Thus, it is important to identify and characterize chaperones for preventing such protein aggregation. In this work, we identified that the NAD+ synthase-nicotinamide mononucleotide adenylyltransferase (NMNAT) 3 from mouse (mN3) exhibits potent chaperone activity to antagonize aggregation of a wide spectrum of pathological amyloid client proteins including α-synuclein, Tau (K19), amyloid β, and islet amyloid polypeptide. By combining NMR spectroscopy, cross-linking mass spectrometry, and computational modeling, we further reveal that mN3 uses different region of its amphiphilic surface near the active site to directly bind different amyloid client proteins. Our work demonstrates a client recognition mechanism of NMNAT via which it chaperones different amyloid client proteins against pathological aggregation and implies a potential protective role for NMNAT in different amyloid-associated diseases.


Antibiotic-induced microbiota depletion in normally-reared adult rats mimics the neuroendocrine effects of early life stress.
Early life stress induced by maternal separation (MS) causes neuroendocrine, behavioral, and metabolic alterations that are related to gut dysbiosis. MS also increases microglial activation and decreases neurogenesis. Whether these long-term alterations are maintained or worsened in the absence of gut microbiota remains unknown. Hence, this study evaluated the effect of MS symptomatology after antibiotic-induced microbiota depletion (AIMD) in adult rats. Control and maternally separated (3 h per day from postnatal day one to 14, MS180) rats were subjected to AIMD for one month, then assessed for behavioral, metabolic, and neuroendocrine responses. Effects of MS180 and AIMD on gut microbiota were confirmed by qPCR. The data indicate that MS180 caused a passive coping strategy in the forced swimming test and decreased hippocampal neurogenesis. In addition, fasting glucose, cholesterol, and corticosterone levels increased, which correlated with a decrease in Lactobacillus spp counts in the caecum. AIMD also increased immobility in the forced swimming test, decreased hippocampal neurogenesis, and augmented corticosterone levels. However, it had no effects on glucose homeostasis or plasma lipid levels. Furthermore, the MS180-induced long-term effects on behavior and neurogenesis were not affected by microbiota depletion. Meanwhile, the metabolic imbalance was partially reversed in MS180 + AIMD rats. These results show that AIMD mimics the behavioral consequences of MS180 but may prevent metabolic imbalance, suggesting that gut dysbiosis could be part of the mechanisms involved in the maintenance of the long-term consequences of early life stress.


Partial rescue of the perfusion deficit area by thrombolysis.
To investigate the evolution of the perfusion deficit area following systemic thrombolysis with recombinant tissue plasminogen activator (rtPA) in a clinical study on acute cerebral ischemia. We performed volumetric measurements of the acute ischemic lesions in MR images of perfusion (TTP, MTT, and rCBV) and in diffusion-weighted (DW) images, as well as the manifest stroke lesions in T2-weighted MR images on day 8. We compared the data of 29 patients who were subjected to systemic thrombolysis with those of 18 patients who were not eligible for thrombolysis. In the treated patients there were prominent MTT/DWI and TTP/DWI mismatches (P < 0.0006). The acute TTP volumes were smaller than the acute MTT volumes, but as large as the T2 lesions on day 8. The MTT/T2 lesion volume reduction was significant (P < 0.03) in patients who received the GPIIb/IIIa receptor antagonist tirofiban (N = 13) in addition to the low-dose rtPA. This corresponded to a greater neurological improvement compared to patients who received rtPA alone (P < 0.05). In contrast, in the nontreated patients the initial MTT and TTP lesion volumes were of similar magnitude and predicted the T2 lesions on day 8. In the treated and nontreated patients the TTP lesion signified the viability threshold of acute ischemia, which corresponded to a rCBF of 25 +/- 11 mL/100 g/min. The perfusion deficit area comprises the ischemic core that is destined to undergo necrosis, and an ischemic rim that is salvageable by systemic thrombolysis.


Plasma concentrations of free and sulfoconjugated dopamine, epinephrine, and norepinephrine in healthy infants and children.
Plasma concentrations of free and sulfoconjugated catecholamines were measured in healthy infants and children under resting conditions. Free norepinephrine and epinephrine levels were up to three times higher in healthy children under 2 years than in adults, even under true resting conditions. In contrast, free dopamine concentrations of all age groups fell within the normal range for adults. The levels of sulfoconjugation were in the adult range.


Current Concepts in the Measurement of Glenohumeral Bone Loss.
The extent of glenohumeral bone loss seen in anterior shoulder dislocations plays a major role in guiding surgical management of these patients. The need for accurate and reliable preoperative assessment of bone loss on imaging studies is therefore of paramount importance to orthopedic surgeons. This article will focus on the tools that are available to clinicians for quantifying glenoid bone loss with a focus on emerging trends and research in order to describe current practices. Recent evidence supports the use of 3D CT as the most optimal method for quantifying bone loss on the glenoid and humerus. New trends in the use of 3D and ZTE MRI represent exciting alternatives to CT imaging, although they are not widely used and require further investigation. Contemporary thinking surrounding the glenoid track concept and the symbiotic relationship between glenoid and humeral bone loss on shoulder stability has transformed our understanding of these lesions and has inspired a new focus of study for radiologists and orthopedist alike. Although a number of different advanced imaging modalities are utilized to detect and quantify glenohumeral bone loss in practice, the current literature supports 3D CT imaging to provide the most reliable and accurate assessments. The emergence of the glenoid track concept for glenoid and humeral head bone loss has inspired a new area of study for researchers that presents exciting opportunities for the development of a deeper understanding of glenohumeral instability in the future. Ultimately, however, the heterogeneity of literature, which speaks to the diverse practices that exist across the world, limits any firm conclusions from being drawn.


The structure of glibenclamide in the solid state.
The structure of glibenclamide, 5-chloro-N-(2-{4-[(cyclohexylamino)carbonyl] aminosulfonyl}phenyl) ethyl)-2-methoxybenzamide, an important antidiabetic drug, has been studied both in solution and in the solid state by a combination of NMR spectroscopy and theoretical calculations. The possibility that glibenclamide suffers a tautomerization under melting to afford a desmotrope was rejected.


Plasma zinc levels in sheep in the peri-parturient period.
No consistent pattern of plasma zinc concentrations was observed in the peri-parturient period in 17 ewes, although large transitory depressions of plasma zinc levels were recorded in five ewes including one non-pregnant control. Sheep with dystocia did not exhibit a fall in plasma zinc similar to that recorded in cattle.


Metabolic effects in the freshwater fish Geophagus iporangensis in response to single and combined exposure to graphene oxide and trace elements.
Graphene oxide (GO) is part of a new set of nanomaterials with particular characteristics related to its nanoscale size. Due to this feature, it presents high reactivity and other contaminants present in the environment could bind to them and affect its intrinsic toxicity. The metabolic effects of such nanomaterials and their combination with two common pollutants, zinc and cadmium, on the freshwater fish Geophagus iporangensis are analyzed. Moreover, metabolic rate and ammonia excretion were used as bioindicators to measure metabolic changes. Fishes were exposed for 24 h in filtered tap water to different concentrations of GO (0.5; 1.0; 2.0 and 4.0 mg L-1), Zn (0.5; 1.0; 2.0; 4.0 and 10.0 mg L-1) and Cd (0.1; 0.5; 1.0; 2.0 and 4.0 mg L-1). Combined effects were verified using the same concentrations of trace elements added to 1.0 mg L-1 of GO. Exposure to GO and Cd resulted in a decrease of metabolic rate in G. iporangensis, by about 30% compared to control means, in the highest concentration tested (4.0 mg L-1). However, zinc exposure in the highest concentration (10 mg L-1) raised metabolic rate to around three times that of the control group. Ammonia excretion was not affected by exposure to GO and Cd. In contrast, exposure to Zn at 10 mg L-1 raised the rate to around 47%. The combined exposure of GO and Zn intensified the effects of the trace element, inducing responses in both biomarkers at lower concentrations and demonstrating that the interaction between elements increases zinc's effects. The combination Cd + GO only affects metabolic rate. Thus, this metabolic rate alone reveals that combined exposure potentiates effects of trace elements on fish metabolism.


Publisher Correction: The TLR7/8 agonist R848 remodels tumor and host responses to promote survival in pancreatic cancer.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Implantable flexible electrodes for functional electrical stimulation.
A manufacturing technology has been developed to fabricate microelectrode systems with reportedly high numbers of electrodes and high reproducibility. The approach leads to flexible microimplants without the need for heavy and large titanium or ceramic housings.


Identification of cuticular protein genes and analysis of their roles in phosphine resistance of the rusty grain beetle Cryptolestes ferrugineus.
The rusty grain beetle, Cryptolestes ferrugineus (Stephens) is one of the most economically important stored grain pests, and it has evolved the high resistance to phosphine. Cuticular proteins (CPs) are the major structural components of insect cuticle, and previous studies have confirmed that CPs were involved in insecticide resistance. However, the CPs of C. ferrugineus are still poorly characterized, and thus we conducted transcriptome-wide identification of CP genes and analyze their possible relationships with phosphine resistance in this pest. In this study, a total of 122 putative CPs were annotated in the C. ferrugineus transcriptome data by blasting with the known CPs of Tribolium castaneum. The analysis of conserved motifs revealed these CPs of C. ferrugineus belonging to 9 different families, including 87 CPR, 13 CPAP1, 7 CPAP3, 3 Tweedle, 1 CPLCA, 1 CPLCG, 5 CPLCP, 2 CPCFC, and 3 CPFL proteins. The further phylogenetic analysis showed the different evolutionary patterns of CPs. Namely, we found some CPs (CPR family) formed species-specific protein clusters, indicating these CPs might occur independently among insect taxa, and while some other CPs (CPAP1 and CPAP3 family) shared a closer correlation based on the architecture of protein domains. Subsequently, the previous RNA-seq data were applied to establish the expression profiles of CPs in a phosphine susceptible and resistant populations of C. ferrugineus, and a large amount of CP genes were found to be over-expressed in resistant insects. Lastly, an up-regulated CP gene (CPR family) was selected for the further functional analysis, and after this gene was silenced via RNA interference (RNAi), the sensitivity to phosphine was significantly enhanced in C. ferrugineus. In conclusion, the present results provided us an overview of C. ferrugineus CPs, and which suggested that the CPs might play the critical roles in phosphine resistance.


Electrocardiographic predictors of coronary heart disease and sudden cardiac deaths in men and women free from cardiovascular disease in the Atherosclerosis Risk in Communities study.
We evaluated predictors of coronary heart disease (CHD) death and sudden cardiac death (SCD) in the Atherosclerosis Risk in Communities (ARIC) study. The study population included 13 621 men and women 45 to 65 years of age free from manifest cardiovascular disease at entry. Hazard ratios from Cox regression with 95% confidence intervals were computed for 18 dichotomized repolarization-related ECG variables. The average follow-up was 14 years. Independent predictors of CHD death in men were TaVR- and rate-adjusted QTend (QTea), with a 2-fold increased risk for both, and spatial angles between mean QRS and T vectors and between Tpeak (Tp) and normal R reference vectors [θ(Rm|Tm) and θ(Tp|Tref), respectively], with a >1.5-fold increased risk for both. In women, independent predictors of the risk of CHD death were θ(Rm|Tm), with a 2-fold increased risk for θ(Rm|Tm), and θ(Tp|Tref), with a 1.7-fold increased risk. Independent predictors of SCD in men were θ(Tp|Tref) and QTea, with a 2-fold increased risk, and θ(Tinit|Tterm), with a 1.6-fold increased risk. In women, θ(Tinit|Tterm) was an independent predictor of SCD, with a >3-fold increased risk, and θ(Rm|Tm) and TV1 were >2-fold for both. θ(Rm|Tm) and θ(Tp|Tref), reflecting different aspects of ventricular repolarization, were independent predictors of CHD death and SCD, and TaVR and TV1 were also independent predictors. The risk levels for independent predictors for both CHD death and SCD were stronger in women than in men, and QTea was a significant predictor in men but not in women.


Characterization of a novel strain of Botryosphaeria dothidea chrysovirus 1 from the apple white rot pathogen Botryosphaeria dothidea.
A novel strain of Botryosphaeria dothidea chrysovirus 1 was identified. It encodes a shortened RNA-dependent RNA polymerase and an elongated coat protein, and it might cause hypovirulence of the host fungal strain.


Evaluation of a balloon dilator before second-trimester abortion by vacuum curettage.
We evaluated the cervical dilator device (CDD), an expanding balloon, as a substitute for laminaria tents before abortion at 13 to 16 1/2 weeks by extraction and vacuum curettage. The CDD was found to be an effective dilator, although it did not appear to have any advantage over laminaria tents when placed overnight and was associated with significant pain upon insertion and an apparent increased risk of endometritis. The present CDD or a subsequent modification may offer advantages over laminaria tents for short-term placement.


Different effects of acarbose and glibenclamide on proinsulin and insulin profiles in people with Type 2 diabetes.
In a double-blind, placebo-controlled study, we compared the effect of acarbose (A) and glibenclamide (G) on post-prandial (pp) and 24-h profiles of proinsulin and insulin. Twenty-seven patients with Type 2 diabetes mellitus insufficiently controlled with diet alone were randomised to receive acarbose, 100 mg thrice daily, glibenclamide, 1 mg thrice daily, or placebo. Before and after 16 weeks of treatment, 24-h profiles of proinsulin, insulin and glucose (fasting, 1 h after breakfast and every 3-h for a 24-h period) were measured under metabolic ward conditions with standardised meals. With acarbose, a reduced 24-h level of proinsulin was observed compared with glibenclamide (AUC 1096 +/- 118 vs. 1604 +/- 174 pmol/l per h, P<0.05) at 16 weeks. The breakfast increment of proinsulin was lower with acarbose than glibenclamide (6.8 vs. 19.3 pmol/l, P<0.05) as was the level at that time (37.3 +/- 5.3 vs. 56.4 +/- 7.5 pmol/l, P<0.05). A lower AUC of insulin after treatment was also observed with acarbose than glibenclamide (7.9 +/- 0.9 vs. 14.8 +/- 4.5 nmol/l per h, P<0.05), as also for 1-h increment (81 +/- 26, vs. 380 +/- 120 pmol/l, P<0.01) and 1-h level (325 +/- 30 vs. 621 +/- 132 pmol/l, P<0.01). Acarbose reduced 1-h breakfast glucose increment (baseline 6.3 +/- 0.6, 16-week 3.5 +/- 0.6 mmol/l, P<0.01) and 1-h glucose level (18.1 +/- 1.1 and 14.5 +/- 1.3 mmol/l, P<0.01), whereas glibenclamide did not (6.6 +/- 0.7 vs. 5.4 +/- 0.6 mmol/l and 18.9 +/- 1.5 vs. 15.3 +/- 1.3 mmol/l). Measurement of circadian excursions of proinsulin and insulin reveals distinct differences in meal-time proinsulin and insulin increment and level between acarbose and glibenclamide whereas fasting levels of these insulin fractions remained unaffected.


Pulmonary cavitation, allergic aspergillosis, asthma and bronchocentric granulomatosis.
Pulmonary cavitation is a rare complication of allergic bronchopulmonary aspergillosis in patients with chronic asthma. Such patients frequently manifest mucoid impaction of a bronchus, "central" saccular bronchiectasis and bronchocentric granulomatosis, a specific microscopic pathologic entity. Early recognition of this syndrome in asthmatic patients and vigorous treatment with steroids may obviate surgery.


Molecular Modeling and Optimization of Type II E.coli l-Asparginase Activity by in silico Design and in vitro Site-directed Mutagenesis.
L-asparaginase (also known as L-ASNase) is a crucial therapeutic enzyme that is widely used in treatment of ALL (acute lymphoblastic leukemia) as a chemotherapeutic drug. Besides, this enzyme is used in the food industry as a food processing reagent to reduce the content of acrylamide in addition to the clinical industry. The improvement of activity and kinetic parameters of the L-ASNase enzyme may lead to higher efficiency resulting in practical achievement. In order to achieve this goal, we chosen glycine residue in position 88 as a potential mutation with advantageous outcomes. In this study, firstly to find the appropriate mutation on glycine 88, various in silico analyses, such as MD simulation and molecular docking, were carried out. Then, the rational design was adopted as the best strategy for molecular modifications of the enzyme to improve its enzymatic properties. Our in silico findings show that the four mutations G88Q, G88L, G88K, and G88A may be able to increase L-ASNase's asparaginase activity. The catalytic efficiency of each enzyme (kcat/Km) is the most important feature for comparing the catalytic activity of the mutants with the wild type form. The laboratory experiments showed that the kcat/Km for the G88Q mutant is 36.32% higher than the Escherichia coli K12 ASNase II (wild type), which suggests that L-ASNase activity is improved at lower concentration of L-ASN. Kinetic characterization of the mutants L-ASNase activity confirmed the high turnover rate (kcat) with ASN as substrate relative to the wild type enzyme. In silico analyses and laboratory experiments demonstrated that the G88Q mutation rather than other mutation (G88L, G88K, and G88A) could improve the kinetics of L-ASNase.


Temoporfin-mediated photodynamic therapy in patients with advanced, incurable head and neck cancer: A multicenter study.
The aim of this study was to confirm the efficacy of temoporfin (Foscan)-mediated photodynamic therapy (PDT) in patients with end-stage head and neck squamous cell carcinoma (HNSCC). Thirty-nine patients with recurring HNSCC lesions ≤10 mm in depth were injected with temoporfin, followed by illumination of the tumor surface. Of 39 treated patients, 19 achieved complete response, 2 partial response, 5 stable disease, 5 had progressive disease, and 8 patients were nonevaluable. Thus, in the per-protocol analysis, the response rate was 68%. Of the treated patients 54% had a response. Median survival was significantly longer for responders (37 months) than for nonresponders (7.4 months). Nine patients were alive at 3.7 to 6.5 years (median, 4.8 years) post-treatment, 7 free of disease. No major toxicities were observed. Patients with advanced HNSCC with lesions ≤10 mm in depth, who have exhausted other treatment options, can achieve significant local control and survival benefit from temoporfin-mediated PDT.


Sugar-derived 2-S-substituted-2H-pyran-3(6H)-ones: synthesis and reactivity.
A practical procedure for the preparation of chiral 2-S-substituted-2H-pyran-3(6H)-ones is described. According to the reaction conditions, 1,5-anhydro-2,3,4-tri-O-acetyl-D-threo-pent-1-enitol (1) reacted with thiophenol under Lewis acid catalysis to afford the polysubstitution product 1,5-anhydro-2,3,4-tri-S-phenyl-2,3,4-trithio-D,L-threo-pent-1-enitol (2) or phenyl 2,4-di-O-acetyl-3-deoxy-1-thio-alpha- and beta-D-glycero-pent-2-enopyranoside (3 and 4, respectively). The iodine-promoted addition of thiophenol or alpha-toluenethiol to 1 gave (2S)-2-phenylthio-2H-pyran-3(6H)-one (5) or its 2-benzylthio analogue 6, but these products showed low enantiomeric excesses (ee approximately 40-60%). However, dihydropyranone 5 with high optical purity (ee>94%) was successfully obtained by treatment of 4 with iodine in acetonitrile. On the other hand, it was established that the benzylthio group of 5 exerts high stereocontrol in reduction and cycloaddition reactions performed on the alpha,beta-unsaturated carbonyl system.


Inhibition of sterol and DNA synthesis in peripheral blood lymphocytes by AY9944.
The effect of the inhibitor of cholesterol synthesis AY9944 on DNA synthesis was tested in phytohemagglutinin stimulated lymphocytes incubated in delipidated serum. AY9944 caused a decrease in cholesterol synthesis from [14C] acetate and an accummulation of [14C] 7-dehydrocholesterol. This inhibition of cholesterol synthesis resulted in an inhibition of DNA synthesis in a dose related manner. Inhibition could be partly reversed by added lipoprotein. Inhibition of DNA synthesis occurred if AY9944 was added at the early stages of blast transformation but not when the response was well established. These findings suggest that some endogenous cholesterol synthesis may be required for the complete phytohemagglutinin stimulation of lymphocytes.


Pathology of modified graft-versus-host disease in bone marrow allografted monkeys treated with antilymphocyte serum.
Lethally irradiated rhesus monkeys were used for bone marrow allografting and autografting. Monkeys receiving allogeneic bone marrow developed acute graft-versus-host reaction (GVHR) and had a mean survival time of 9.1 days as compared to autografted monkeys which survived above 500 days. Treatment with antilymphocyte sera (ALS) before allografting modified the GVHR and extended the survival time to an average of 43 days. Histologically, such animals showed evidence of "chronic" GVHR and septicemia secondary to a lack in lymphoreticular recovery. Subsequently, severe GI-tract infections followed which usually served as portal of entry for septicemia.


Ectopic meningioma of the maxillary sinus.
This paper presents a rare case of meningioma arising in the right maxillary sinus of a 23-year-old Tanzanian male. It appeared as a painless swelling of the right side of the face. The diagnosis was made after histologic examination of the specimen removed by lateral rhinotomy.


Internal hernia trends following gastric bypass surgery.
Internal hernia is a well-known complication of laparoscopic Roux-en-Y gastric bypass (LRYGB), with reported rates ~ 5% within three months to three years after surgery. Internal hernia through a mesenteric defect can lead to small bowel obstruction. Mesenteric defects began to be more routinely closed, often considered standard practice by 2010. To our knowledge, there are no large population-based studies looking at rates of internal hernia post-LRYGB. This study utilizes a statewide database to characterize the trends of internal hernia post-LRYGB over the last two decades in multiple centers. LRYGB procedure records between January 2005 and September 2015 were extracted from the New York SPARCS database. Exclusion criteria included age < 18, in-hospital deaths, bariatric revision procedures, and internal hernia repair during the same hospitalization as LRYGB. Time to internal hernia was calculated from initial LRYGB hospital stay to admission date of the first internal hernia repair record. A multivariable proportional sub-distribution hazards model was utilized to analyze the trend of internal hernia incidence within three-year post-LRYGB. 46,918 patients were identified between 2005 and 2015, with 2950 (6.29) undergoing internal hernia repair post-LRYGB by the end of 2018. The cumulative incidence of internal hernia repair at the 3rd-year post-LRYGB was 4.80% (95% CI: 4.59%-5.02%). By the end of the 13th year, the longest follow-up period, the cumulative incidence was 12.00% (95% CI: 11.30%-12.70%). Overall, there was a decreasing trend over time of undergoing internal hernia repair within three-year post-LRYGB (HR = 0.94, 95% CI: 0.93-0.96), after adjusting for confounding factors. This multicenter study maintains the rate of internal hernia following LRYGB reported in smaller studies and provides a longer follow-up period demonstrating decreasing occurrences of internal hernia after bypass as a function of year of index operation. This data is important as internal hernia continues to be a complication post-LRYGB.