Nickel(II) in chelate N2O2 environment. DFT approach and in-depth molecular orbital and configurational analysis.
The O-N-N-O-type tetradentate ligands H2S,S-eddp (H2S,S-eddp stands for S,S-ethylenediamine-N,N'-di-2-propionic acid) and H2edap (H2edap stands for ethylenediamine-N-acetic-N'-3-propionic acid) and the corresponding novel octahedral nickel(II) complexes have been prepared and characterized. N2O2 ligands coordinate to the nickel(II) ion via four donor atoms (two deprotonated carboxylate atoms and two amine nitrogens) affording octahedral geometry in the case of all investigated Ni(II) complexes. A six coordinate, octahedral geometry has been verified crystallographically for the s-cis-[Ni(S,S-eddp)(H2O)2] complex. Structural data correlating similarly chelated Ni(II) complexes have been used to carry out an extensive configuration analysis. Molecular mechanics and Density Functional Theory (DFT) have been used to model the most stable geometric isomer, yielding, at the same time, significant structural and spectroscopic (TDDFT) data. The results from density functional studies have been compared to X-ray data. Natural Bond Orbital (NBO) and Natural Energetic Decomposition Analysis (NEDA) have been done for the [Ni(edda-type)(H2O)(2-n)] and nH2O fragments. Molecular orbital analysis (MPA) is given as well. The infra-red and electronic absorption spectra of the complexes are discussed in comparison to the related complexes of known geometries.


A proteomics strategy for the analysis of bacterial biodegradation pathways.
Bacterial biodegradation (bioremediation) is the use of microorganisms to break down organic materials into simpler compounds; it plays a pivotal role in the clean-up of hazardous wastes in the environment. Following the completion of genome sequencing in bacteria capable of biodegradation, functional genomic studies have played a major role in obtaining information on bacterial biodegradation pathways. Novel proteomics technologies have recently been developed to make it possible to analyze global protein expression. Proteomics can also provide important information on the life cycle, regulation, and post-translational modification of proteins induced under specific conditions. Proteomics technologies have been applied to the comprehensive study of bacterial biodegradation. In this paper, we introduce the proteomics technologies applicable to bacterial biodegradation studies, review the results of the proteomics analysis of representative biodegrading bacteria, and discuss the potential use of proteomics technologies in future biodegradation studies.


Behavioral Therapy for Children with Avoidant/Restrictive Food Intake Disorder Dependent on Tube or Oral Enteral Nutrient Formula: A Feasibility Study.
In children with eating disorders, nutritional status and growth may depend on enteral nutrient formula. Ultimately, its goal is to introduce or reintroduce oral feeding. Japanese research on the treatment of tube or oral formula-dependent children is scarce. This study determined the feasibility of behavioral therapy for children with avoidant/restrictive food intake disorder and dependency on the tube or oral enteral nutrient formula in Japan. Medical records of children diagnosed with this disorder, dependent on the tube or oral enteral nutrient formula and who had received behavioral therapy intervention to withdraw from the formula were retrospectively investigated. We collected their characteristics at first visit and the caloric percentage from oral food intake six months after starting the treatment. In total, four patients (age range: 2-5 years) participated in this study. The feeding routes employed before the intervention were a nasogastric tube for one patient, a gastrostomy bottom for the other patient, and oral formula for the remaining patients (i.e., two children). At the sixth month of the behavioral treatment, none of the patients needed the formula, and the caloric percentage of required nutrition from oral food intake was 100%. Our data demonstrate that this behavioral therapy is feasible for children with avoidant/restrictive food intake disorder dependent on the tube or oral formula in Japan.


Pseudocapacitive Sodium Storage in Mesoporous Single-Crystal-like TiO2-Graphene Nanocomposite Enables High-Performance Sodium-Ion Capacitors.
Sodium-ion capacitors can potentially combine the virtues of high power capability of conventional electrochemical capacitors and high energy density of batteries. However, the lack of high-performance electrode materials has been the major challenge of sodium-based energy storage devices. In this work, we report a microwave-assisted synthesis of single-crystal-like anatase TiO2 mesocages anchored on graphene as a sodium storage material. The architecture of the nanocomposite results in pseudocapacitive charge storage behavior with fast kinetics, high reversibility, and negligible degradation to the micro/nanostructure. The nanocomposite delivers a high capacity of 268 mAh g-1 at 0.2 C, which remains 126 mAh g-1 at 10 C for over 18 000 cycles. Coupling with a carbon-based cathode, a full cell of sodium-ion capacitor successfully demonstrates a high energy density of 64.2 Wh kg-1 at 56.3 W kg-1 and 25.8 Wh kg-1 at 1357 W kg-1, as well as an ultralong lifespan of 10 000 cycles with over 90% of capacity retention.


Immunomodulatory Effects of Subacute Inhalation Exposure to Copper Oxide Nanoparticles in House Dust Mite-Induced Asthma.
It has been shown that inhalation exposure to copper oxide nanoparticles (CuO NPs) results in pulmonary inflammation. However, immunomodulatory consequences after CuO NP inhalation exposure have been less explored. We tested the effect of CuO NP aerosols on immune responses in healthy, house dust mite (HDM) asthmatic, or allergen immunotherapy (AIT)-treated asthmatic mice (BALB/c, females). The AIT consisted of a vaccine comprising HDM allergens and CpG-loaded nanoparticles (CpG NPs). AIT treatment involved mice being immunized (via subcutaneous (sc) injection; 2 doses) while concomitantly being exposed to CuO NP aerosols (over a 2 week period), starting on the day of the first vaccination. Mice were then sensitized twice by sc injection and subsequently challenged with HDM extract 10 times by intranasal instillation. The asthmatic model followed the same timeline except that no immunizations were administered. All mice were necropsied 24 h after the end of the HDM challenge. CuO NP-exposed healthy mice showed a significant decrease in TH1 and TH2 cells, and an elevation in T-bet+ Treg cells, even 40 days after the last exposure to CuO NPs. Similarly, the CuO NP-exposed HDM asthma model demonstrated decreased TH2 responses and increased T-bet+ Treg cells. Conversely, CuO NP inhalation exposure to AIT-treated asthmatic mice resulted in an increase in TH2 cells. In conclusion, immunomodulatory effects of inhalation exposure to CuO NPs are dependent on immune conditions prior to exposure.


Properties of purified CYP2R1 in a reconstituted membrane environment and its 25-hydroxylation of 20-hydroxyvitamin D3.
Recent studies indicate that CYP2R1 is the major 25-hydroxylase catalyzing the first step in vitamin D activation. Since the catalytic properties of CYP2R1 have been poorly studied to date and it is a membrane protein, we examined the purified enzyme in a membrane environment. CYP2R1 was expressed in E. coli and purified by nickel affinity- and hydrophobic interaction-chromatography and assayed in a reconstituted membrane system comprising phospholipid vesicles plus purified human NADPH-P450 oxidoreductase. CYP2R1 converted vitamin D3 in the vesicle membrane to 25-hydroxyvitamin D3 [25(OH)D3] with good adherence to Michaelis-Menten kinetics. The kinetic parameters for 25-hydroxylation of vitamin D3 by the two major vitamin D 25-hydroxylases, CYP2R1 and CYP27A1, were examined in vesicles under identical conditions. CYP2R1 displayed a slightly lower kcat than CYP27A1 but a much lower Km for vitamin D3, and thus an overall 17-fold higher catalytic efficiency (kcat/Km), consistent with CYP2R1 being the major vitamin D 25-hydroxylase. 20-Hydroxyvitamin D3 [20(OH)D3], the main product of vitamin D3 activation by an alternative pathway catalyzed by CYP11A1, was metabolized by CYP2R1 to 20,25-dihydroxyvitamin D3 [20,25(OH)2D3], with catalytic efficiency similar to that for the 25-hydroxylation of vitamin D3. 20,25(OH)2D3 retained full, or somewhat enhanced activity compared to the parent 20(OH)D3 for the inhibition of the proliferation of melanocytes and dermal fibroblasts, with a potency comparable to 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. The 20,25(OH)2D3 was also able to act as an inverse agonist on retinoic acid-related orphan receptor α, like its parent 20(OH)D3. Thus, the major findings of this study are that CYP2R1 can metabolize substrates in a membrane environment, the enzyme displays higher catalytic efficiency than CYP27A1 for the 25-hydroxylation of vitamin D, it efficiently hydroxylates 20(OH)D3 at C25 and this product retains the biological activity of the parent compound.


Direct chemical method for preparing 2,3-epoxyamides using sodium chlorite.
A direct method for preparing 2,3-epoxyamides from tertiary allylamines via a tandem C-H oxidation/double bond epoxidation using sodium chlorite is reported. Apparently, the reaction course consists of two steps: (i) allylic oxidation of the starting allylamine to corresponding unsaturated allylamide with sodium chlorite followed by (ii) epoxidation of the allylamide to the 2,3-epoxyamide mediated by hypochlorite ion, which is formed in situ by reduction of sodium chlorite. The reaction conditions tolerate the presence of free hydroxyl groups and typical functional groups such as TBS, aryl, alkyl, allyl, acetyl, and benzyl groups; however, when an activated aromatic ring (e.g., sesamol) is present in the substrate, the use of a scavenger is necessary.


Metastasis to the thyroid gland from primary breast cancer presenting as diffuse goiter: A case report and review of literature.
Metastasis to the thyroid gland (TM) from primary breast cancer is uncommon and usually presents as thyroid nodules; however, diffuse goiter without thyroid nodules is the first sign of TM in rare cases. Skip metastases (SMs) to the lymph nodes in breast cancer, defined as discontiguous higher-level metastases in the absence of lower levels of contiguous metastases, have been reported in the contralateral cervical area of the primary tumor site in rare cases. A 49-year-old previously healthy Chinese woman was diagnosed with right lateral invasive ductal carcinoma and underwent neoadjuvant chemotherapy treatment and bilateral mastectomy with axillary lymph node dissection. No malignancy of the left breast or axillary or distant metastases were identified preoperatively. However, enlarged left cervical lymph nodes were detected 36 mo after surgery, and rapidly enlarging thyroid glands without nodules were detected 42 mo after surgery. Fine-needle aspiration cytology was performed on the left cervical lymph nodes and left lobe of the thyroid, which were both revealed to contain metastases from the primary breast cancer. Additionally, the immunostaining profiles changed in the process of metastases. The patient was discharged with the NP (vinorelbine and cisplatin) regimen for subsequent treatment, and stable disease was determined when the curative effect was evaluated. Diffuse goiter may be the first sign of TM, and enlarged lymph nodes in the contralateral cervical area may be SMs of primary breast cancer.


Greater fluid loss does not fully explain the divergent hemodynamic balance mediating postexercise hypotension in endurance-trained men.
Following exercise, mean arterial pressure (MAP) is reduced ~5-10 mmHg from preexercise baseline. In nonendurance-trained males, postexercise hypotension results from peripheral vasodilation not offset by increased cardiac output (CO). By contrast, postexercise hypotension occurs through a reduction in CO from preexercise baseline in endurance-trained males. The reason(s) explaining these divergent responses remain unknown. Exercise at fixed percentage of peak oxygen consumption (V̇o2peak) is associated with a greater rate of metabolic heat production in trained individuals and therefore elevated sweat rates, both when compared with untrained individuals. We hypothesized that greater fluid loss would explain the postexercise reduction in CO of endurance-trained males. Twelve endurance-trained males (Trained: V̇o2peak, 64 ± 5 ml O2·kg-1·min-1) cycled for 60 min at 60% V̇o2peak (Trained60%). On separate days, 12 nonendurance trained males (Untrained: V̇o2peak, 49 ± 3 ml O2·kg-1·min-1) cycled at 1) 60% V̇o2peak (Untrained60%), and 2) a rate of heat production equivalent to that achieved by the Trained group (UntrainedMatched). Fluid loss was similar between Trained60% (-1.32 ± 0.20 kg) and UntrainedMatched (-1.32 ± 0.23 kg; P = 0.99) but was greater in these conditions relative to Untrained60% (-0.95 ± 0.11 kg; both P < 0.01). During the final 30 min of postexercise supine recovery, MAP was similarly reduced by 5 ± 2 mmHg in all three conditions ( P = 0.91). The reduction in MAP was mediated by a 0.5 ± 0.3 l/min reduction in CO from baseline in Trained60% ( P = 0.01). In contrast, CO returned to baseline following exercise during UntrainedMatched and Untrained60% (both P ≥ 0.30). These data demonstrate that greater fluid loss does not fully explain the divergent postexercise hemodynamic responses observed in trained relative to untrained males. NEW & NOTEWORTHY Even when matched for exercise-induced fluid loss, cardiac output was decreased in trained males but returned to baseline following exercise in their untrained counterparts. However, as per our hypothesis, reductions in stroke volume were similar between groups. This suggests that exercise-induced fluid loss is an important determinant of the stroke volume response during recovery but factors affecting heart rate such as exercise intensity and/or heat stress are also important determinants of postexercise hemodynamics.


[Effect of trimebutine on cholinergic transmission in neurons of the inferior mesenteric ganglion of the rabbit].
We analyzed the effects of trimebutine on the synaptic activity of neurons of the rabbit inferior mesenteric ganglion, using intracellular recording techniques. The synaptic activity was produced by subthreshold stimuli (0.5 Hz) applied individually, on lumbar splanchnic and lumbar colonic nerves. These stimuli triggered cholinergic responses corresponding to fast excitatory postsynaptic potentials. In 8 of 20 neurones tested trimebutine (10(-6) g/ml) produced an inhibition of excitatory postsynaptic potentials, without any change in the resting membrane potential. In 6 of 20 neurons tested, trimebutine produced, successively, an early facilitation followed by a late inhibition of excitatory postsynaptic potentials. Both effects occurred without change in the resting membrane potential. The inhibitory and facilitatory effects of trimebutine were accompanied, by an increase and a decrease in the number of failures of nerve stimulation respectively. These results indicate that inhibitory and facilitatory effects of trimebutine correspond respectively to a decrease and an increase in the amount of acetylcholine released from presynaptic nerve terminals originating from the spinal cord and the distal colon.


5-Nitroindole as an universal base analogue.
4-, 5- and 6-Nitroindole have been investigated and compared with 3-nitropyrrole as universal bases in oligodeoxynucleotides. Of these the 5-nitroindole derivative was found to be superior giving higher duplex stability, and behaving indiscriminately towards each of the four natural bases in duplex melting experiments. 3-Nitropyrrole, whilst not discriminating between the natural bases, was found to lead to considerable destabilisation of the duplexes, particularly when multiple substitutions were made, in contrast to the 5-nitroindole nucleoside.


Subjectivity of the Anomalous Sense of Self Is Represented in Gray Matter Volume in the Brain.
The self includes complicated and heterogeneous functions. Researchers have divided the self into three distinct functions called "agency," "ownership," and "narrative self". These correspond to psychiatric symptoms, behavioral characteristics and neural responses, but their relationship with brain structure is unclear. This study examined the relationship between the subjectivity of self-related malfunctions and brain structure in terms of gray matter (GM) volume in 96 healthy people. They completed a recently developed self-reported questionnaire called the Embodied Sense of Self Scale (ESSS) that measures self-related malfunctions. The ESSS has three subscales reflecting the three distinct functions of the self. We also determined the participants' brain structures using magnetic resonance imaging (MRI) and voxel-based morphometry (VBM). Multiple regression analysis revealed a significant negative correlation between ownership malfunction and the insular cortex GM volume. A relationship with brain structure could thus only be confirmed for the ESSS "ownership" subscale. This finding suggests that distinct brain structures feel ownership and that the ESSS could partly screen for distinct brain structures.


Identification and in silico analysis of helical lipid binding regions in proteins belonging to the amphitropic protein family.
The role of protein-lipid interactions is increasingly recognized to be of importance in numerous biological processes. Bioinformatics is being increasingly used as a helpful tool in studying protein-lipid interactions. Especially recently developed approaches recognizing lipid binding regions in proteins can be implemented. In this study one of those bioinformatics approaches specialized in identifying lipid binding helical regions in proteins is expanded. The approach is explored further by features which can be easily obtained manually. Some interesting examples of members of the amphitropic protein family have been investigated in order to demonstrate the additional features of this bioinformatics approach. The results in this study seem to indicate interesting characteristics of amphitropic proteins and provide insight into the mechanistic functioning and overall understanding of this intriguing class of proteins. Additionally, the results demonstrate that the presented bioinformatics approach might be either an interesting starting point in protein-lipid interactions studies or a good tool for selecting new focus points for more detailed experimental research of proteins with known overall protein-lipid binding abilities.


Alternative medicine use in dialysis patients: potential for good and bad!
Although alternative medicines are widely used within the general population, the extent of their use within the dialysis population is unknown. It is possible that dialysis patients may be more likely to turn towards alternative therapies in view of the chronicity of their disease. In addition, this particular patient population could be at an increased risk of toxicity from these therapies due to an absence of renal excretion. A detailed assessment of complementary and alternative medicine use in our dialysis patients revealed that 18% of our patients had used or were using some form of alternative medicine therapy. An additional 63% of our patients, however, were willing to use a complementary or alternative medication. Our results suggest that hemodialysis patients are extremely receptive to the use of such therapies and are therefore exposed to all their potential benefit and harm.


Superior short-term learning effect of visual and sensory organisation ability when sensory information is unreliable in adolescent rhythmic gymnasts.
Rhythmic gymnasts specialise in dynamic balance under sensory conditions of numerous somatosensory, visual, and vestibular stimulations. This study investigated whether adolescent rhythmic gymnasts are superior to peers in Sensory Organisation test (SOT) performance, which quantifies the ability to maintain standing balance in six sensory conditions, and explored whether they plateaued faster during familiarisation with the SOT. Three and six sessions of SOTs were administered to 15 female rhythmic gymnasts (15.0 ± 1.8 years) and matched peers (15.1 ± 2.1 years), respectively. The gymnasts were superior to their peers in terms of fitness measures, and their performance was better in the SOT equilibrium score when visual information was unreliable. The SOT learning effects were shown in more challenging sensory conditions between Sessions 1 and 2 and were equivalent in both groups; however, over time, the gymnasts gained marginally significant better visual ability and relied less on visual sense when unreliable. In conclusion, adolescent rhythmic gymnasts have generally the same sensory organisation ability and learning rates as their peers. However, when visual information is unreliable, they have superior sensory organisation ability and learn faster to rely less on visual sense.


A unified mechanism for the stability of surface nanobubbles: contact line pinning and supersaturation.
In this paper, we apply the molecular dynamics simulation method to study the stability of surface nanobubbles in both pure fluids and gas-liquid mixtures. First, we demonstrate with molecular simulations, for the first time, that surface nanobubbles can be stabilized in superheated or gas supersaturated liquid by the contact line pinning caused by the surface heterogeneity. Then, a unified mechanism for nanobubble stability is put forward here that stabilizing nanobubbles require both the contact line pinning and supersaturation. In the mechanism, the supersaturation refers to superheating for pure fluids and gas supersaturation or superheating for the gas-liquid mixtures, both of which exert the same effect on nanobubble stability. As the level of supersaturation increases, we found a Wenzel or Cassie wetting state for undersaturated and saturated fluids, stable nanobubbles at moderate supersaturation with decreasing curvature radius and contact angle, and finally the liquid-to-vapor phase transition at high supersaturation.


Synergistic Effect of Doripenem and Cefotaxime to Inhibit CTX-M-15 Type β-Lactamases: Biophysical and Microbiological Views.
CTX-M-15 type β-lactamase has the ability to hydrolyse cefotaxime, a third generation cephalosporin. The infections caused by multidrug resistant strains, especially CTX-M-15 producing strains are being treated with carbapenem group of β-lactam antibiotics. The objective of the study was to know if cefotaxime in combination with doripenem (carbapemen antibiotic) at very low concentration, inhibits the CTX-M-15 producing bacterial strains. blaCTX-M-15 gene was cloned to express CTX-M-15 enzyme and construct CTX-M-15 producing strain. The clone carrying CTX-M-15 was found susceptible to doripenem. Doripenem and CTX-M-15 binding was an endothermic and spontaneous process leading to change in polarity in the micro-environment and conformational changes of enzyme as shown by fluorescence, UV and CD spectroscopic study. The catalytic efficiency of CTX-M-15 enzyme was reduced to about 15.86% when it was treated with doripenem along with cefotaxime (in 5 times molar ratio each of doripenem and cefotaxime w.r.t CTX-M-15), as compared to the studies where enzyme's efficiency was increased by 33% when treated with cefotaxime alone. Hence, doripenem in combination with cefotaxime reduces enzyme's efficiency to hydrolyse cefotaxime by about 48%. FIC study showed that doripenem paired with cefotaxime showed synergistic effect against CTX-M-15 producing bacterial strain. The study concludes that doripenem at very low concentration (25 nM), induces such a structural changes in CTX-M-15 which reduced enzyme's activity to hydrolyse cefotaxime. Hence, the synergistic use of doripenem and cefotaxime plays a significant role in inhibiting the efficiency of CTX-M-15 type β-lactamase, and may provide an alternative approach to reduce the resistance against the cephalosporin type antibiotics.


Anti-K562 cell monoclonal antibody RTF8X recognizes tumor-associated antigen of erythroid lineage.
A new anti-K562 cell monoclonal antibody, RTF8X, a cytotoxic IgM, recognized a surface antigen on erythroblasts from patients with erythroleukemia and polycythemia vera. RTF8X, which is highly specific to K562 cells, did not react with the other 14 hematopoietic cell lines and the seven nonhematopoietic cell lines. RTF8X antigen was not detected in normal peripheral blood, but was found in less than 1% of normal marrow cells. RTF8X did not inhibit in vitro colony formation of CFU-E and BFU-E in a complement-dependent cytotoxicity assay. Cell-sorting analysis showed that, morphologically, the RTF8X-positive marrow cells from the patients and normal volunteers contained more than 60% erythroblasts and that CFU-E and BFU-E were not demonstrated in cells with RTF8X antigen. Enzyme treatment suggested that RTF8X antigen was a sialoglycolipid. These results indicate that RTF8X may recognize the surface antigen found increasingly in association with tumors of erythroid lineage. RTF8X should be useful for studies of erythroid differentiation and proliferation in patients.


Expression of Bcl-2 and Bax in rhesus monkey testis during germ cell apoptosis induced by testosterone undecanoate.
Apoptosis occurs spontaneously during spermatogenesis and can be induced by androgen withdrawal. However, the molecular events governing apoptosis have not been characterized. To study the molecular mechanism of apoptosis induced by a high dose of testosterone undecanoate (TU), the authors examined the temporal changes in proapoptotic Bax and antiapoptotic Bcl-2 in TU-treated monkey testes. Apoptotic cells were identified in tissue sections by in situ end labeling of fragmented DNA. The results showed that a great deal of the apoptotic cells occurred in the testes on day 30 after TU injection and that the dominant apoptotic germ cells are spermatocytes and spermatids. The expression of Bcl-2 and Bax was assessed by immunohistochemical method and Western blot. As compared with that of normal testes, the levels of Bcl-2 protein increased significantly from 7 to day 14 while that of Bax protein was almost unchanged in the testes from day 7 up to day 60 after TU treatment. Bcl-2 was localized to the spermatids in the normal testes and temporarily distributed in both the cytoplasm and nucleus of those cell types susceptible to TU-induced apoptosis on day 14 after TU injection. Therefore, it is suggested that Bax may not play a role in initiating germ cell apoptosis induced by TU injection and that the evaluation in Bcl-2 expression may represent a survival mechanism for the remaining germ cell.


Late-stage synthesis and application of photoreactive probes derived from direct benzoylation of heteroaromatic C-H bonds.
A C-H functionalization strategy for the expedient access to photoreactive chemical probes of commonly found heterocyclic fragments or drug molecules of pharmaceutical relevance is described. A series of aryl glyoxylic acid reagents featuring pendant alkyne or azide clickable handles have been developed for application in the radical-mediated appendage of benzoyl fragments onto simple heteroaromatic fragments, as well as more complex drug-like compounds. This unprecedented strategy of chemical probe synthesis allows for direct access to photoreactive chemical probes without any requirement of fragment pre-functionalization or significant synthetic re-evaluation.


Pharmaceutical-Grade Rigosertib Is a Microtubule-Destabilizing Agent.
We recently used CRISPRi/a-based chemical-genetic screens and cell biological, biochemical, and structural assays to determine that rigosertib, an anti-cancer agent in phase III clinical trials, kills cancer cells by destabilizing microtubules. Reddy and co-workers (Baker et al., 2020, this issue of Molecular Cell) suggest that a contaminating degradation product in commercial formulations of rigosertib is responsible for the microtubule-destabilizing activity. Here, we demonstrate that cells treated with pharmaceutical-grade rigosertib (>99.9% purity) or commercially obtained rigosertib have qualitatively indistinguishable phenotypes across multiple assays. The two formulations have indistinguishable chemical-genetic interactions with genes that modulate microtubule stability, both destabilize microtubules in cells and in vitro, and expression of a rationally designed tubulin mutant with a mutation in the rigosertib binding site (L240F TUBB) allows cells to proliferate in the presence of either formulation. Importantly, the specificity of the L240F TUBB mutant for microtubule-destabilizing agents has been confirmed independently. Thus, rigosertib kills cancer cells by destabilizing microtubules, in agreement with our original findings.


A Singlet-Diradical Co(III)-Dimer as a Nonvolatile Resistive Switching Device: Synthesis, Redox-Induced Interconversion, and Current-Voltage Characteristics.
Herein we report a ligand-centered redox-controlled strategy for the synthesis of an unusual binuclear diradical cobalt(III) complex, [Co2III(L•3-)2] (1), featuring two three-electron reduced trianionic monoradical 2,9-bis(phenyldiazo)-1,10-phenanthroline ligands (L•3-) and two intermediate-spin cobalt(III) centers having a Co-Co bond. Controlled ligand-centered oxidation of 1 afforded two mononuclear complexes, [CoII(L•-)(L0)]+ ([3])+ and [CoII(L0)2]2+ ([2]2+), which upon further ligand-centered reduction yielded a di-azo-anion diradical complex, [CoII(L•-)2] (4). In complex 1, two three-electron reduced di-azo-anion monoradical ligands (L•3-) bridge two intermediate Co(III) centers at a distance of 2.387(2) Å, while upon oxidation, one of the coordinating azo-arms of L becomes pendent, and in complexes [2]2+, [3]+, and 4, two tetradentate ligands coordinate a single Co(II) center in a tridentate meridional fashion with one uncoordinated azo-arm from each of the ligands. In the presence of reducing agents, the monomers [2]2+, [3]+, and 4 undergo ligand-centered reduction to form azo-anion radicals, and the otherwise pendent azo-arms in the presence of cobalt(II)-salts like Co(ClO4)2 or CoCl2 bind the second Co(II)-ion; further internal electron transfer from the cobalt center to the arylazo backbone produces the binuclear complex 1. Spectroscopic analysis, DFT studies, and control experiments were performed to understand the electronic structures and the ligand-centered redox-controlled interconversion. The application of complex 1 as a molecular memory device (memristor) was also explored. Complex 1 showed encouraging results as a memristor with a current ON/OFF ratio > 104 and is highly promising for resistive RAM/ROM applications.


Comparison of the flexural strength of six reinforced restorative materials.
This study calculated the flexural strength for six reinforced restorative materials and demonstrated that flexural strength values can be determined simply by using physical parameters (diametral tensile strength and Young's modulus values) that are easily determined experimentally. A one-way ANOVA analysis demonstrated a statistically significant difference between the two reinforced glass ionomers and the four composite resin materials, with the composite resin being stronger than the glass ionomers.


Metabolic study of methylstenbolone in horses using liquid chromatography-high resolution mass spectrometry and gas chromatography-mass spectrometry.
Methylstenbolone (2,17α-dimethyl-5α-androst-1-en-17β-ol-3-one) is a synthetic anabolic and androgenic steroid (AAS) sold as an oral 'nutritional supplement' under the brand names 'Ultradrol', 'M-Sten' and 'Methyl-Sten'. Like other AASs, methylstenbolone is a prohibited substance in both human and equine sports. This paper describes the studies of the in vitro and in vivo metabolism of methylstenbolone in horses using LC/HRMS, GC/MS and GC/MS/MS. Phase I in vitro metabolic study of methylstenbolone was performed using homogenised horse liver. Hydroxylation was the only biotransformation observed. Six in vitro metabolites were detected including four mono-hydroxylated metabolites, namely 16α/β-hydroxymethylstenbolone (M1a, M1b), 20-hydroxymethylstenbolone (M1c), 6-hydroxymethylstenbolone (M1d), and two dihydroxylated methylstenbolone metabolites (M2c-M2d). An in vivo experiment was carried out using two retired thoroughbred geldings. Each horse was administered with 100 mg methylstenbolone supplement by stomach tubing daily for three consecutive days. Methylstenbolone and 14 metabolites were detected in the post-administration urine samples. The proposed in vivo metabolites included 16α/β-hydroxymethylstenbolone (M1a, M1b), 20-hydroxymethylstenbolone (M1c), two dihydroxylated methylstenbolone (M2a, M2b), 17-epi-methylstenbolone (M3), methasterone (M4), 2,17-dimethylandrostane-16,17-diol-3-one (M5), dihydroxylated and reduced methylstenbolone (M6), 2α,17α-dimethylandrostane-3α,17β-diol (M7), 2,17-dimethylandrostane-3,16,17-triol (M8a-M8c) and 2,17-dimethylandrostane-2,3,16,17-tetraol (M9), formed from hydroxylation, reduction and epimerisation. Methylstenbolone and ten of its metabolites could be detected in post-administration plasma samples. The highest concentration of methylstenbolone detected in urine was about 10-36 ng/mL at 3-4 h after the last administration, while the maximum concentration in plasma was about 0.4-0.7 ng/mL at 1 h after the last administration. For controlling the misuse of methylstenbolone, M8c and M9 gave the longest detection time in urine, while M4, M5 and M6 were the longest detecting analytes in plasma. They could be detected for up to 5 and 4.5 days respectively in urine and plasma. Apart from 16α/β-hydroxymethylstenbolone (M1a, M1b), the methylstenbolone metabolites reported herein have never been reported before.


On-chip three-dimensional tumor spheroid formation and pump-less perfusion culture using gravity-driven cell aggregation and balanced droplet dispensing.
This paper presents a spheroid chip in which three-dimensional (3D) tumor spheroids are not only formed by gravity-driven cell aggregation but also cultured at the perfusion rates controlled by balanced droplet dispensing without fluidic pumps. The previous spheroid chips require additional off-chip processes of spheroid formation and extraction as well as bulky components of fluidic pumps. However, the present spheroid chip, where autonomous medium droplet dispensers are integrated on a well array, achieves the on-chip 3D tumor spheroid formation and perfusion culture using simple structure without bulky fluidic pumps. In the experimental study, we demonstrated that the spheroid chip successfully forms 3D tumor spheroids in the wide diameter range of 220 μm-3.2 mm (uniformity > 90%) using H358, H23, and A549 non-small cell lung cancer cells. At the pump-less perfusion culture (Q = 0.1-0.3 μl/min) of spheroids, the number of H358 cells in the spheroid increased up to 50% from the static culture (Q = 0 μl/min) and the viability of the cultured cells also increased about 10%. Therefore, we experimentally verified that the perfusion environment created by the spheroid chip offers a favourable condition to the spheroids with high increase rate and viability. The present chip achieves on-chip 3D tumor spheroid formation and pump-less perfusion culture with simple structure, thereby exhibiting potential for use in integrated in-vivo-like cell culture systems.


Mapping of Ebola virus spillover: Suitability and seasonal variability at the landscape scale.
The unexpected Ebola virus outbreak in West Africa in 2014 involving the Zaire ebolavirus made clear that other regions outside Central Africa, its previously documented niche, were at risk of future epidemics. The complex transmission cycle and a lack of epidemiological data make mapping areas at risk of the disease challenging. We used a Geographic Information System-based multicriteria evaluation (GIS-MCE), a knowledge-based approach, to identify areas suitable for Ebola virus spillover to humans in regions of Guinea, Congo and Gabon where Ebola viruses already emerged. We identified environmental, climatic and anthropogenic risk factors and potential hosts from a literature review. Geographical data layers, representing risk factors, were combined to produce suitability maps of Ebola virus spillover at the landscape scale. Our maps show high spatial and temporal variability in the suitability for Ebola virus spillover at a fine regional scale. Reported spillover events fell in areas of intermediate to high suitability in our maps, and a sensitivity analysis showed that the maps produced were robust. There are still important gaps in our knowledge about what factors are associated with the risk of Ebola virus spillover. As more information becomes available, maps produced using the GIS-MCE approach can be easily updated to improve surveillance and the prevention of future outbreaks.


Heavy Rainfall Impacts on Trihalomethane Formation in Contrasting Northwestern European Potable Waters.
There is emerging concern over the impact of extreme events such as heavy rainfall on the quality of water entering the drinking water supply from aboveground sources, as such events are expected to increase in magnitude and frequency in response to climate change. We compared the impact of rainfall events on streamwater quality in four contrasting upland (peatland and mineral soil) and lowland agricultural catchments used to supply drinking water in France (Brittany) and the United Kingdom (North Wales) by analyzing water samples collected before, during, and after specific events. At all four streams, heavy rainfall led to a considerable rise in organic matter concentration ranging from 48 to 158%. Dissolved organic carbon (DOC) quality, as determined using specific ultraviolet absorbance, changed consistently at all sites during rainfall events, with a greater proportion of aromatic and higher molecular weight compounds following the onset of rainfall. However, the change in DOC quality and quantity did not significantly alter the trihalomethane formation potential. We observed small increases in trihalomethane (THM) generation only at the Welsh peatland and agricultural sites and a small decrease at the Brittany agricultural site. The proportion of brominated THMs in chlorinated waters was positively correlated with bromide/DOC ratio in raw waters for all sites and hydrological conditions. These results provide a first indication of the potential implications for surface-based drinking water resources resulting from expected future increases in rainfall event intensity and extension of dry periods with climate changes.


Injury mechanisms and severity in narrow offset frontal impacts.
Current standard frontal crash tests include full frontal or 40% offset. Frontal impacts with offsets less than 40% and corner impacts have received little attention. Because of the limited engagement of vehicle structures that would permit less energy dissipation, these crashes have the potential for severe trauma to the near-side occupant. Narrow offset and corner-impact crashes under a frontal impact classification were analyzed using data obtained from the United States Department of Transportation National Highway Traffic Safety Administration crash databases: Crash Injury Research and Engineering Network (CIREN) and the National Automotive Sampling System (NASS) 2000-2006. A subset of crashes that could be defined clearly as narrow offset crashes were then examined. The Collision Damage Classification (CDC) classification was used to obtain only crashes with a "FLEE" code for drivers and a "FREE" code for right front passengers. These codes were used to separate out crashes that had bumper and lower vehicle engagement and less than 41 cm of front vehicle damage. The NASS data indicated that corner impacts ("Y" and "Z" type) and narrow overlap ("L" and "R" type) constitute 26 % each of all frontal impacts. When occupants were severely injured in frontal crashes 20 % of the occupants were in "L" and "R" type crashes and 31 % were in "Y" and "Z" type crashes. The percentage of near side "FLEE" and "FREE" crash occupants with severe injuries by body region demonstrated that head, thorax, upper, and lower extremity injuries dominate the data set. For the 71 CIREN cases, lower extremity injuries dominated. The injury severity score did not correlate well with assumed severity parameters like extent of crush and delta-V, however, crashes with extent zones 2-5 had a greater percentage of occupants with chest and spine injuries than crashes with extent zones 6-9. Vehicle rotation after front impact in extent zone 2-5 crashes may be influential in airbag effectiveness.


Cerebral and spinal air embolism following percutaneous nephrolithotomy.
We present a case report of cerebral and spinal air embolism following percutaneous nephrolithotomy in a patient without evidence of intra-cardiac defects or prepulmonary A-V shunts. The position of the patient during the incidence determined the site of eventual lodgement of air emboli in the arterial circulation. We suspect that the time of onset of symptoms following the procedure may be the clue to the path followed by air emboli.


Necessity for and control of dental treatment in HIV infected children. Inter-professional relationship between dentist and paediatrician.
HIV infected children frequently suffer from buccal-dental lesions needing dental treatment. This treatment should improve their systemic affection, localised pathology, psychological and affective state and their general quality of life. Hardly any of these children are ever treated; sometimes because of lack of family motivation (the most frequent cause) and others because of the lack of a Paediatric Dental Unit in the hospitals they attend. For this reason we present here two cases of HIV infected children, with HIV infected mothers, who, thanks to the relationship between the Paediatric and Dental Units of the hospital, have had access to dental treatment for multiple caries and candidiasis. An anatopathological diagnosis of gingival inflammation, which was also infected by candida, was also carried out. The treatment included extraction of teeth, pulpotomy, pulpectomy and the fitting of prostheses. It should be made clear that a good inter-professional relationship is needed and it must also be taken into account the great difficulty that is encountered when trying to make this group of patients understand the need for, and benefits of, dental treatment.


The Diagnostic Utility and Clinical Implications of Wrist MRI in the Pediatric Population.
Unexplained wrist pain is a common presentation in children. To our knowledge, no studies have explored the clinical utility of magnetic resonance imaging (MRI) in the diagnostic workup of pediatric patients. We retrospectively reviewed 307 consecutive wrist MRIs ordered at a tertiary-care pediatric hospital. Demographic data and the indication for imaging were recorded and grouped into admitting categories. The final impression of each MRI was scored with regard to potential impact on future treatment (0 = normal, 1 = minimal, 2 = moderate, 3 = high). Patients who went on to wrist surgery within 1 year were noted. In our cohort, 27% of all studies were normal, including 34% of those with pain. Although pain was the most common category, MRI was most useful in the delineation of a mass/cyst, evaluating for infection and evaluating arthropathy. Compared with all other categories, patients with pain were 3.6 times more likely to have a normal study and 4.6 times more likely to have a clinical score less than or equal to 1. Given an admitting diagnosis of pain, females were 1.7 times more likely to present for an MRI and 2.4 times more likely to have a normal MRI. The Spearman correlation revealed no linear relationship between age and MRI outcome. In all, 13% of patients went on to have surgery within 1 year of MRI. At our pediatric institution, the majority of wrist MRIs were ordered for wrist pain. Given our data, wrist MRI is not an ideal screening tool in children, particularly in those with wrist pain, and should only be used to exclude or confirm a specific diagnosis.


Note: Detector collimators for the nanoscale ordered materials diffractometer instrument at the Spallation Neutron Source.
Five neutron collimator designs were constructed and tested at the nanoscale ordered materials diffractometer (NOMAD) instrument. Collimators were made from High Density PolyEthylene (HDPE) or 5% borated HDPE. In all cases, collimators improved the signal to background ratio and reduced detection of secondary scattering. In the Q-range 10-20 Å(-1), signal to background ratio improved by factors of approximately 1.6 and 2.0 for 50 and 100 mm deep collimators, respectively. In the Q-range 40-50 Å(-1), the improvement factors were 1.8 and 2.7. Secondary scattering as measured at Q ∼ 9.5 Å(-1) was significantly decreased when the collimators were installed.


Downregulation of the basic peroxidase isoenzyme from Zinnia elegans by gibberellic acid.
Hypocotyl formation during the epigeal germination of seedlings is under strict hormonal regulation. In a 3 d old Zinnia elegans seedling system, gibberellic acid (GA(3)) exerts an opposite effect to that exerted by light on hypocotyl photomorphogenesis because GA(3) promotes an etiolated-like growth with an inhibition of radial (secondary) growth. For this reason, the effect of GA(3) on the basic peroxidase isoenzyme from Z. elegans (ZePrx), an enzyme involved in hypocotyl lignin biosynthesis, was studied. The results showed that GA(3) reduces ZePrx activity, similarly to the way in which it reduces seedling secondary growth. This hormonal response is supported by the analysis of the ZePrx promoter, which contains four types of GA(3)-responsive cis-elements: the W Box/O2S; the Pyr Box; the GARE; and the Amy Box. Taken together, these results suggest that ZePrx is directly regulated by GA(3), with this effect matching the inhibitory effect of GA on the hypocotyl secondary growth.


Influence of abomasal infusion of glucose or partially hydrolyzed starch on pancreatic exocrine secretion in beef steers.
Five crossbred steers (348 +/- 12 kg) fitted with a pancreatic pouch draining the main pancreatic duct and duodenal re-entrant and abomasal infusion cannulas were used in a 5 x 5 Latin square design to determine the influence of postruminal carbohydrate source and level on pancreatic exocrine secretion in beef steers. Abomasal infusion treatments (250 mL infused/h) were water (control), 20 g/h glucose, 40 g/h glucose, 20 g/h starch hydrolysate (SH), and 40 g/h SH. Infusion periods were 8 d with 3 to 4 d of rest between periods. Pancreatic juice was collected for 6 h on d 8 of each collection period. Every 30 min a 10% subsample was composited and frozen and the remainder was infused into the duodenum via the reentrant cannula. Abomasal infusion of glucose or SH increased (P < 0.10) total secretion of pancreatic juice and decreased (P < 0.10) secretion of alpha-amylase activity. Abomasal carbohydrate infusion did not influence total secretion of protein, trypsin activity, or chymotrypsin activity. This experiment indicates that increasing postruminal glucose or SH decreases pancreatic alpha-amylase secretion.


Separation and Preconcentration of Nickel(II) from Drinking, Spring, and Lake Water Samples with Amberlite CG-120 Resin and Determination by Flame Atomic Absorption Spectrometry.
In this study, Amberlite CG-120 adsorbent was used for the separation/preconcentration of Ni(II) ions in commercial drinking, spring and lake water samples before detection by flame atomic absorption spectrometry. Various optimization parameters for Ni(II) determination, such as pH, eluent type and concentration, sample and eluent flow rates, amount of adsorbent, were investigated to obtain better sensitivity, accuracy, precision and quantitative recovery. Furthermore, the interference effects of some ions on the recovery efficiency of Ni(II) were also investigated. The optimum experimental parameters were obtained in the case of pH 1; 5 mL of 4 mol L-1 HCl for eluent and 0.3 g for the adsorbent amount. The limit of detection was found to be 0.58 μg L-1 and linearity ranged from 5 to 50 μg L-1. The accuracy of the method was tested by the certified reference material of TMDA-70.2 Ontario Lake Water at a 95% confidence level.


Distribution of Subchondral Bone Puncture Strength in the Talus and Tibial Plafond: A Biomechanical Study.
We investigated talus and tibial plafond subchondral bone puncture strength based on surface location. Puncture tests of the subchondral bone were performed in 9 equal zones on the articular surface of 12 cadaver specimens aged 49.1 years (range, 36-56 years). Compressive load was applied through a microfracture awl at 2 mm/min. Puncture strength was defined as the first load drop in load-deflection curves. In the talus, zone 1 (215 ± 91 N) and zone 2, the anterior medial and anterior middle zones, had significantly greater puncture strength than zones 7, 8, and 9, the posterior medial, middle, and lateral zones (104 ± 43 N, 115 ± 43 N, and 102 ± 35 N, respectively; P < .001). In the tibial plafond, zone 3, the anterior-lateral zone, and zone 7, the posterior medial zone, had significantly greater strength than zone 8, the posterior middle zone (202 ± 72 N, 206 ± 121 N, and 112 ± 65 N, respectively; P < .001) These results suggest that the subchondral bone is significantly weaker to penetrative force in the posterior region than in the anterior region of the talar dome and of the tibial plafond. These findings may have implications for microfracture awl design and for understanding the complex anatomy and physiology of the ankle joint. Controlled laboratory study.


Supramolecular redox-responsive substrate carrier activity of a ferrocenyl Janus device.
Supramolecular Janus compounds have recently attracted increasing attention owing to their dynamic reversible properties, distinct topological structures, and remarkable physicochemical characteristics, e.g., amphiphilicity, heterofunctionality, and high-density of terminal groups. Herein, a new redox-responsive supramolecular Janus device was designed and synthesized involving β-cyclodextrin and 2-fold ferrocene host-guest interactions. The complex formation was analyzed via one-dimensional 1H NMR and two-dimensional Nuclear Overhauser Enhancement Spectroscopy. FeCl3 and ascorbic acid were used as oxidation and reduction triggers, respectively, to modulate the self-assembly behavior in water through complexation/dissociation of β-cyclodextrin inclusion compounds resulting from redox-conversion of the ferrocenyl guest moieties. The redox-responsiveness of the obtained supramolecular micelles was studied via scanning electron microscopy and dynamic light scattering. Substrate-loading ability of the supramolecular micelles was confirmed with Rhodamine B, and the oxidation of ferrocenyl groups led to the release of the loaded cargos. The present work illustrates a valuable design example of supramolecular Janus systems using the host-guest complexation between β-cyclodextrin and ferrocenyl structures. The present supramolecular micelle may be used as a promising molecular vehicle for application in the field of stimuli-responsive drug delivery.


Principles and nursing management of anticoagulation.
Anticoagulant drugs are widely used in hospital and community settings. Anticoagulation is the first-line treatment for venous thromboembolism, and anticoagulant drugs have an important role in the treatment and prevention of blood clots. However, maintaining the equilibrium between clotting and bleeding can be challenging and anticoagulants have been identified as a class of drug associated with preventable patient harm. Direct oral anticoagulants (DOACs) have become the first-line treatment for many patients requiring an anticoagulant, removing the burden of frequent tests and the many food and drug interactions associated with vitamin K antagonists such as warfarin sodium. However, DOACs have increased the complexity of decision-making regarding treatment, which also increases the risk of drug errors. This article discusses the uses, modes of action and potential side effects of anticoagulants, to improve nurses' understanding and enable them to have an active role in limiting the risk of harm from these drugs.


[The sensitized rate of preformed antibodies in the candidate of renal transplant recipient].
It is important to examine the presence of preformed antibodies in hemodialysis patients before renal transplantation, because it causes the graft rejection in organ transplantation. In the 239 hemodialysis patients we analysed 3 preformed antibodies divided TW, BW and BC. Sensitized rate (TW+, BW+ and BC+) was 0% at no blood transfusions (BT), 9.3% at 1-5 BT and 28.9% at 6 more BT. The number of blood transfusions increased the sensitized rate (TW). Sensitized rate (BW+, BC+) was 6.2% at no BT, 9.3% at 1-5BT and 12.0% at 6 more BT, while sensitized rate (BC+) was 33.3% at no BT, 26.7% at 1-5BT and 14.5% at 6 more BT. There was no correlation between the number of blood transfusions and the sensitized rate (BW+, BC+). At one year after the first check, 69.2% of TW positive patients was converted to TW negative and 53.8% of BW positive patients was converted to BW negative each. Further studies of the preformed antibodies are required to assess the clinical course of long surviving patients.


Flexible light manipulation in non-Hermitian frequency Su-Schrieffer-Heeger lattice.
Recently, studies on non-Hermitian topologic physics have attracted considerable attention. The non-Hermitian skin effect (NHSE), as a remarkable phenomenon in the non-Hermitian lattice, has been demonstrated in coupled ring resonators and photonic mesh lattices. However, there is a scarcity of work on the realization of NHSEs in synthetic dimensions, owing to inaccessible anisotropic coupling. This limits the potential for exploring non-Hermitian topologic physics in on-chip integrated optical systems. In this work, we implement a non-Hermitian Su-Schrieffer-Heeger topologic insulator in the synthetic frequency dimension, and the NHSE and topologic edge state are manifested. Furthermore, we demonstrate that the exotic chiral Zener tunneling can also be realized. Our system provides a versatile platform to explore and exploit non-Hermitian topologic physics on a chip and can have impacts on flexible light manipulation in frequency domains.


Ion-pulling simulations provide insights into the mechanisms of channel opening of the skeletal muscle ryanodine receptor.
The type 1 ryanodine receptor (RyR1) mediates Ca2+ release from the sarcoplasmic reticulum to initiate skeletal muscle contraction and is associated with muscle diseases, malignant hyperthermia, and central core disease. To better understand RyR1 channel function, we investigated the molecular mechanisms of channel gating and ion permeation. An adequate model of channel gating requires accurate, high-resolution models of both open and closed states of the channel. To this end, we generated an open-channel RyR1 model using molecular simulations to pull Ca2+ through the pore constriction site of a closed-channel RyR1 structure determined at 3.8-Å resolution. Importantly, we find that our open-channel model is consistent with the RyR1 and cardiac RyR (RyR2) open-channel structures reported while this paper was in preparation. Both our model and the published structures show similar rotation of the upper portion of the pore-lining S6 helix away from the 4-fold channel axis and twisting of Ile-4937 at the channel constriction site out of the channel pore. These motions result in a minimum open-channel pore radius of ∼3 Å formed by Gln-4933, rather than Ile-4937 in the closed-channel structure. We also present functional support for our model by mutations around the closed- and open-channel constriction sites (Gln-4933 and Ile-4937). Our results indicate that use of ion-pulling simulations produces a RyR1 open-channel model, which can provide insights into the mechanisms of channel opening complementing those from the structural data.


Modeling of cell viability and specific alcohol productivity.
A model has been proposed to correlate two viability tests and specific alcohol productivity of a yeast culture. Two viable populations with different activity are assumed to coexist. The difference between two populations is the ability to reproduce. Satisfactory agreement between the model and the experimental data has been observed.


Healthcare-associated infections and antimicrobial resistance in severe acquired brain injury: a retrospective multicenter study.
Recent studies underscore that healthcare-associated infections (HAIs) and multidrug-resistant (MDR) HAIs affect rehabilitation outcomes and hospital length of stay (LOS) for severe acquired brain injury (sABI). This study aimed to estimate HAI incidence in different sABI rehabilitation settings and determine risk factors and HAI impact on neuromotor and cognitive recovery. We conducted a retrospective multicenter study in two semi-intensive units (SICUs), two high-specialty post-acute units (PAUs), and one long-term care (LTC) rehabilitation facility. Data extraction was performed by experienced clinicians, using a structured Excel file and they agreed upon criteria for case definitions of healthcare. The main outcome measures were the HAI and MDR HAI incidence and the LOS, the functional recovery was measured using the Level of Cognitive Functioning and Disability Rating Scale. There were 134 sABI participants. The calculation of the probability level was adjusted for three pairwise comparisons among settings (0.05/3 = 0.017). The HAI and MDR HAI incidences were significantly higher in SICU (3.7 and 1.3 per 100 person-days) than in other settings (LTC: 1.9, p = 0.034 and 0.5, p = 0.026; PAU: 1.2, p < 0.001 and 0.3, p < 0.001). HAI and MDR HAI risk variables included older age, an increased number of devices, and carbapenemase-producing Enterobacteriaceae (CPE) colonization, while a high prealbumin plasma value seemed to have a protective effect. HAIs are related to longer LOS, and colonization is associated with poor prognosis and poor functional outcomes with reduced ability to achieve the cognitive capacity of self-care, employability, and independent living. The need to ensure the protection of non-colonized patients, especially those with severe disabilities on admission, is highlighted.


Pathogenesis of bacterial exacerbations of COPD.
Acute exacerbations are significant events in the course of COPD. The pathogenesis of exacerbations was poorly understood, specifically, the role of bacteria was highly controversial. Recent observations have demonstrated that bacterial infection is involved in about half of the exacerbations. The predominant mechanism of bacterial exacerbation in COPD appears to be acquisition of new strains of bacterial pathogens from the environment that are able to establish infection in the tracheobronchial tree in COPD because of compromised innate lung defenses. These pathogens interact with airway cells, elicit an inflammatory response, which underlies the pathophysiology and symptoms characteristic of exacerbation. An immune response that can be mucosal, systemic or both develops to the infecting bacterial strain. This immune response contains the infectious process, could eradicate the infecting pathogen and prevent re-infection with the same strain. However, because of antigenic diversity among bacterial strains, this immunity tends to be strain-specific rather than widely protective. Other mechanisms, including increase in bacterial load and interaction with other etiologies such as viruses, also could contribute to bacterial exacerbations. Improved understanding of the host-pathogen interaction in the airways in COPD will lead to novel approaches to prevention and treatment of exacerbations.


Comparison of brain volume abnormalities between ADHD and conduct disorder in adolescence.
Previous studies of brain structure abnormalities in conduct disorder and attention-deficit/hyperactivity disorder (ADHD) samples have been limited owing to cross-comorbidity, preventing clear understanding of which structural brain abnormalities might be specific to or shared by each disorder. To our knowledge, this study was the first direct comparison of grey and white matter volumes in diagnostically "pure" (i.e., no comorbidities) conduct disorder and ADHD samples. Groups of adolescents with noncormobid conduct disorder and with noncomorbid, combined-subtype ADHD were compared with age- and sex-matched controls using DARTEL voxel-based analysis of T1-weighted brain structure images. Analysis of variance with post hoc analyses compared whole brain grey and white matter volumes among the groups. We included 24 adolescents in each study group. There was an overall 13% reduction in grey matter volume in adolescents with conduct disorder, reflecting numerous frontal, temporal, parietal and subcortical deficits. The same grey matter regions typically were not abnormal in those with ADHD. Deficits in frontal lobe regions previously identified in studies of patients with ADHD either were not detected, or group differences from controls were not as strong as those between the conduct disorder and control groups. White matter volume measurements did not differentiate conduct disorder and ADHD. Our modest sample sizes prevented meaningful examination of individual features of ADHD or conduct disorder, such as aggression, callousness, or hyperactive versus inattentive symptom subtypes. The evidence supports theories of frontotemporal abnormalities in adolescents with conduct disorder, but raises questions about the prominence of frontal lobe and striatal structural abnormalities in those with noncomorbid, combined-subtype ADHD. The latter point is clinically important, given the widely held belief that ADHD is associated with numerous frontal lobe structural deficits, a conclusion that is not strongly supported following direct comparison of diagnostically pure groups. The results are important for future etiological studies, particularly those seeking to identify how early expression of specific brain structure abnormalities could potentiate the risk for antisocial behaviour.


[Study on correlation between trace elements and active ingredient in glycyrrhizae radix et rhizome].
To clear the kinds of trace elements which is closely related with the active ingredient, proclaim the effects of trace elements on the quality of the Glycyrrhizae Radix et Rhizoma, provide the theoretical foundation to the further quality control of cultivation, take the advantage of the HPLC to determine the contents of glycyrrhizic acid and the liquiritin according to Chinese pharmacopoeia, use the inductively coupled plasma mass spectrometry to test the contents of Cu, Zn, Mn, Pb, Se, Cd, Ni, La, Na, Cr, M, Fe, Ca, Al, K, Sr, then, use SPSS statistical software for active ingredient and trace elements Correlation Analysis. The result of correlation analysis showed that Liquiritin contents of Glycyrrhizae Radix et Rhizoma have strong a positive correlation with the Mn, Pb contents, well, have a negative correlation with the Cu, Na contents; Glycyrrhizic acid contents showed a positive correlation with Mg, Cd, La contents, however, it showed a negative correlation with K, Fe contents. Comprehensive analysis of the results of the study, a preliminary thought that the active ingredient of Glycyrrhizae Radix et Rhizoma closely related with the trace elements, but the exact conclusion still need further study concentration-response relationship analysis.


[Effects of high sodium diet on dopaminergic mechanism in normal and hypertensive subjects].
To investigate the effects of dietary sodium on the peripheral dopaminergic mechanism, changes of unconjugated plasma dopamine(DA) and its related humoral factors were studied in 8 patients with essential hypertension(EH) and 8 age-matched normal controls(N) while they were receiving ordinary meals (Na, 130-180 mEq daily) followed by higher sodium (250-300 mEq daily) diets for a week. Plasma and urinary DA, norepinephrine(NE) and epinephrine(E) were measured by the highly sensitive COMT-mediated radioenzymatic procedure, which permits an accurate estimation of plasma DA as low as 5-6 pg/ml. Under high sodium diets, blood pressure and heart rate were not changed significantly in N and EH subjects. Urinary NE and E tended to decrease, while urinary DA increased significantly in both groups of subjects (p less than 0.05). There was a significant correlation between urinary sodium and DA (r = 0.590, p less than 0.001), but plasma DA failed to correlate significantly to urinary sodium or DA in all subjects. Plasma NE and E tended to decrease in both N and EH subjects, while plasma DA increased significantly (p less than 0.05) in EH from 7.2 +/- 0.8 pg/ml [mean +/- SEM] to 9.3 +/- 1.0 and slightly in N from 9.1 +/- 1.8 to 11.2 +/- 1.3. Plasma renin activity(PRA) and plasma aldosterone(PAC) were invariably decreased in all subjects, while plasma prolactin(PRL) remained unchanged. A significant correlation was observed between plasma DA and NE under ordinary meals (r = 0.733, p less than 0.01), but this correlation disappeared under high sodium diets. Plasma DA showed an inverse correlation to PAC (r = 0.351, p less than 0.05) under both dietary conditions. Upright posture induced a significant rise (p less than 0.05) in NE, E, DA, PRA and PAC with ordinary meals, but the responses of NE and PAC were apparently attenuated with high sodium diets. An intravenous injection of metoclopramide (MCP, 10 mg), a DA receptor antagonist, provoked a slight rise in plasma NE and DA with ordinary meals, of which responses were further enhanced with high sodium diets. MCP induced a definite rise in PAC and PRL in all subjects under both dietary conditions (p less than 0.01), while plasma E and PRA remained unchanged after MCP challenge. The results lend support to the view that unconjugated plasma DA could be a useful marker of peripheral dopaminergic activity, which might be a physiological regulator responsible for the suppression of aldosterone secretion and sympathetic nerve activity observed during high sodium intake.


18F-FDG PET/CT and primary hepatic MALT: a case series.
Primary hepatic mucosa-associated lymphoid tissue (MALT) lymphoma is an extremely rare disease and its glucidic metabolic behavior is not clear. We retrospectively analyzed five patients with histological diagnosis of primary hepatic MALT lymphoma who underwent twelve 18F-FDG PET/CT. All staging 18F-FDG PET/CT were positive showing 18F-FDG uptake (average SUVmax was 5.62 ± 1.6) at the corresponding liver lesion. 18F-FDG PET/CT also was useful in evaluating the complete metabolic response after chemotherapy in three patients and radiotherapy in two. Besides, in one patient 18F-FDG PET/CT detected disease relapse during follow-up. Despite the low number of patients, our case series shows the 18F-FDG avidity of hepatic MALT and the possible role of 18F-FDG PET/CT in the management of these patients, both for staging, treatment response evaluation and restaging. Further studies are needed to confirm our results.


Simultaneous measurement of the viability, aggregation, and live and dead adherence of Streptococcus crista, Streptococcus mutans and Actinobacillus actinomycetemcomitans in human saliva in relation to indices of caries, dental plaque and periodontal disease.
Salivary proteins have multiple functions and many share similar functions, which may be why it has been difficult to relate variations in their concentrations to oral health and ecology. An alternative is to focus on variations in the major functions of saliva. An hydroxyapatite-coated microplate model has been developed that simultaneously measures saliva-promoted bacterial viability, bacterial aggregation, and live and dead bacterial adherence, while simulating oral temperature and shearing forces from swallowing. That model was applied to resting whole and stimulated parotid saliva from 149 individuals, using representative strains of Streptococcus crista, S. mutans, and Actinobacillus actinomycetemcomitans. Two major factors were defined by multivariate analysis (this was successful only for whole-saliva). One factor was correlated with aggregation, live adherence and dead adherence for all three strains; the other was correlated with total viability of all three strains. Participants were grouped <25th percentile and >75th percentile for each factor. Those groups were compared for clinical indices of oral health. Caries scores were significantly lower in those with high scores for aggregation-adherence, regardless of whether total viability scores were low or high. Live bacteria always predominated on surfaces when live and dead adherence scores were expressed as ratios. However, participants with high scores for aggregation-adherence showed significantly more dead adherent bacteria than those with low scores (these ratios were uncorrelated with total viability). This finding may indicate that extreme differences in the ability to kill bacteria on surfaces can influence caries risk.


IGF-I in human breast cancer: low differentiation stage is associated with decreased IGF-I content.
Few investigations on the potential role of IGF-I in human breast cancer have used morphological criteria, and the data presented on the localisation of IGF-I are controversial. Moreover, little information exists on a potential correlation between local IGF-I and the grade of malignancy or prognostic factors. Therefore, we investigated the immunohistochemical localisation of IGF-I in specimens of human breast cancer tumours of the ductal type, graded as G1/G2 (well-/moderately differentiated, n=115) and G3 (poorly differentiated, n=28). IGF-I immunoreactivity was quantified using a scaling from no (-) to numerous (+++) IGF-I-immunoreactive cells. From 29 of the G1/G2 and 17 of the G3 tumours IGF-I was also measured by RIA. Cytosolic oestrogen receptor (ER) and progesterone receptor (PR) levels, and S-phase fraction were established and related to the number of IGF-I-immunoreactive cells. IGF-I immunoreactivity occurred predominantly in ductal epithelial cells. Of G3 tumours, 57% exhibited IGF-I immunoreactivity as compared with 84% of G1/G2 tumours. Correspondingly, the amount of IGF-I measured by RIA was significantly lower in G3 tumours (6.9+/-0.9 ng/g wet weight) than in G1/G2 tumours (10.5+/-1.1 ng/g wet weight) (P=0.031). G1/G2 tumours exhibited a higher percentage of IGF-I-immunoreactive cells (16% -, 23% +, 41% ++, 20% +++) than G3 tumours (43% -, 37% +, 12% ++, 8% +++). When comparing the - with the +++ G1/G2 tumours, the frequency of IGF-I-immunoreactive cells was related significantly to the ER (P<0.016) and the PR (P<0.008) levels. In G1/G2 and G3 tumours, the ER and PR levels increased with the amount of IGF immunoreactivity while the S-phase fraction increased with decreasing IGF-I content. In 25% of the specimens, IGF-I immunoreactivity occurred in stromal cells, but there was no obvious difference between the different types of tumours. The survival of the G1/G2 tumour patients increased with increasing numbers of IGF-I-immunoreactive cells. It is concluded that IGF-I is associated with the more-differentiated type of epithelial cells and that increasing dedifferentiation goes along with decreased IGF-I content. Thus, the presence of IGF-I immunoreactivity in breast cancer epithelial cells indicates a lower degree of malignancy than the lack of IGF-I.


Minocycline protects, rescues and prevents knockdown transgenic parkin Drosophila against paraquat/iron toxicity: Implications for autosomic recessive juvenile parkinsonism.
Autosomal recessive Juvenile Parkinsonism (AR-JP) is a chronic, progressive neurodegenerative disorder caused by mutation in the PARKIN gene, and invariably associated with dopaminergic (DAergic) neuronal loss and brain iron accumulation. Since current medical therapy is symptomatic and lacks significant disease-modifying effects, other treatment approaches are urgently needed it. In the present work, we investigate the role of minocycline (MC) in paraquat (PQ)/iron-induced neurotoxicity in the Drosophila TH>parkin-RNAi/+ (w[*]; UAS-parkin-RNAi; TH-GAL4) fly and have shown the following: (i) MC increased life span and restored the locomotor activity of knockdown (KD) transgenic parkin flies in comparison with the control (vehicle) group; (ii) MC at low (0.1 and 0.3mM) and middle (0.5mM) concentrations protected, rescued and prevented KD parkin Drosophila against PQ toxicity. However, MC at high (1mM) concentration aggravated the toxic effect of PQ; (iii) MC protected and rescued DAergic neurons against the PQ toxic effect according to tyrosine hydroxylase (TH)>green-fluorescent protein (GFP) reporter protein microscopy and anti-TH Western blotting analysis; (iv) MC protected DAergic neurons against PQ/iron toxicity; (v) MC significantly abridged lipid peroxidation (LPO) in the protection, rescue and prevention treatment in TH>parkin-RNAi/+ flies against PQ or iron alone or combined (PQ/iron)-induced neuronal oxidative stress (OS). Our results suggest that MC exerts neuroprotection against PQ/iron-induced OS in DAergic neurons most probably by the scavenging activity of reactive oxygen species (ROS), and by chelating iron. Therefore, MC might be a potential therapeutic drug to delay, revert, or prevent AR-JP.


Hydrolysate from eggshell membrane ameliorates intestinal inflammation in mice.
Inflammatory bowel diseases (IBD) comprises of ulcerative colitis (UC) and Cohn's disease (CD) as two main idiopathic pathologies resulting in immunologically mediated chronic inflammatory conditions. Several bioactive peptides and hydro lysates from natural sources have now been tested in animal models of human diseases for potential anti-inflammatory effects. Eggshell membrane (ESM) is a well-known natural bioactive material. In this study, we aim to study the anti-inflammatory activity of ESM hydro lysate (AL-PS) in vitro and in vivo. In vitro, AL-PS was shown to inhibit pro-inflammatory cytokine IL-8 secretion. In vivo treatment with AL-PS was shown to reduce dextran sodium sulphate (DSS)-induced weight loss, clinical signs of colitis and secretion of interleukin (IL)-6 (p < 0.05). In addition, treatment with AL-PS also attenuated the severity of intestinal inflammation via down-regulation of IL-10 an anti-inflammatory cytokine. This validates potential benefits of AL-PS as a novel preventative target molecule for treatment of IBD.


Methods of detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis in periodontal microbiology, with special emphasis on advanced molecular techniques: a review.
Certain specific bacterial species from the subgingival biofilm have demonstrated aetiological relevance in the initiation and progression of periodontitis. Among all the bacteria studied, three have shown the highest association with destructive periodontal diseases: Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg) and Tannerella forsythensis (Tf). Therefore, the relevance of having accurate microbiological diagnostic techniques for their identification and quantification is clearly justified. To evaluate critically all scientific information on the currently available microbial diagnostic techniques aimed for the identification and quantification of Aa, Pg and Tf. Bacterial culturing has been the reference diagnostic technique for many years and, in fact, most of our current knowledge on periodontal microbiology derives from cultural data. However, the advent of new microbial diagnostics, mostly based on immune and molecular technologies, has not only highlighted some of the shortcomings of cultural techniques but has also allowed their introduction as easy and available adjunct diagnostic tools to be used in clinical research and practice. These technologies, mostly polymerase chain reaction (PCR), represent a field of continuous development; however, we still lack the ideal diagnostic to study the subgingival microflora. Qualitative PCR is still hampered by the limited information provided. Quantitative PCR is still in development; however, the promising early results reported are still hampered by the high cost and the equipment necessary for the processing. Quantitative PCR technology may have a major role in the near future as an adjunctive diagnostic tool in both epidemiological and clinical studies in periodontology. However, culture techniques still hold some inherent capabilities, which makes this diagnostic tool the current reference standard in periodontal microbiology.


Ab initio self-consistent x-ray absorption fine structure analysis for metalloproteins.
X-ray absorption fine structure is a powerful tool for probing the structures of metals in proteins in both crystalline and noncrystalline environments. Until recently, a fundamental problem in biological XAFS has been that ad hoc assumptions must be made concerning the vibrational properties of the amino acid residues that are coordinated to the metal to fit the data. Here, an automatic procedure for accurate structural determination of active sites of metalloproteins is presented. It is based on direct multiple-scattering simulation of experimental X-ray absorption fine structure spectra combining electron multiple scattering calculations with density functional theory calculations of vibrational modes of amino acid residues and the genetic algorithm differential evolution to determine a global minimum in the space of fitting parameters. Structure determination of the metalloprotein active site is obtained through a self-consistent iterative procedure with only minimal initial information.


Selection of Reference Genes for RT-qPCR Analysis Under Extrinsic Conditions in the Hawthorn Spider Mite, Amphitetranychus viennensis.
Hawthorn spider mite, Amphitetranychus viennensis Zacher, is an economically important arthropod pest for fruit trees and woody ornamental plants. Extensive and repetitive use of synthetic acaricides has led to the development of resistance in A. viennensis. To understand the molecular basis of pesticide resistance, and to develop genetic-based control alternatives (e.g., RNAi-based biopesticides), a standardized protocol for real-time quantitative reverse transcription PCR (RT-qPCR) is needed. In the proceeding phase of this research, we screened for the internal references for RT-qPCR analysis from a pool of A. viennensis housekeeping genes under the intrinsic conditions, including developmental stage, sex, and diapause. Here, we continued our efforts to search for the reference genes under an array of extrinsic conditions, including temperature, humidity, photoperiod, host plant, and dietary RNAi. The stability of these candidate reference genes was investigated using geNorm, NormFinder, BestKeeper, and ΔCt method, respectively. Finally, RefFinder, a statistical platform integrating all four algorisms, provided a comprehensive list of genes for each extrinsic condition: (1) EF1A, α-tubulin and Actin3 were the best candidates for temperature, (2) GAPDH, 18S, and Actin3 were the most stable genes for humidity, (3) V-ATPase B, Actin3, and 18S were the top reference genes for photoperiod, (4) GAPDH, V-ATPase B, and α-tubulin were recommended for host plants, and (5) GAPDH, V-ATPase B, and RPS9 were the top choices for dietary RNAi. Overall, V-ATPase B, GAPDH, and Actin3 were the most commonly selected reference genes in A. viennensis regardless of the experimental conditions, including both intrinsic and extrinsic. Information present here lays the foundation for the genomic and functional genomic research in A. viennensis.


[Morphology of bovine adenovirus type I].
Electron-microscopic studies were carried out on a concentrated and purified adenovirus type I. It was found that its morphologic aspects were characteristic of the adenovirus group. In the preparations of this virus there were large amounts of incomplete virions and soluble components--hexons, groups of hexons and pentons. Some of the virions were grouped in p-crystal structures or were released as grouped in cell vacuoles. The hexons presented a six-angle contour and a central cavity. The pentons possessed fibre components that were comparatively long. The presence of free pentons spoke of the existence of incomplete hemagglutinins.


Combined Effects of Electric Stimulation and Microgrooves in Cardiac Tissue-on-a-Chip for Drug Screening.
Animal models and traditional cell cultures are essential tools for drug development. However, these platforms can show striking discrepancies in efficacy and side effects when compared to human trials. These differences can lengthen the drug development process and even lead to drug withdrawal from the market. The establishment of preclinical drug screening platforms that have higher relevancy to physiological conditions is desirable to facilitate drug development. Here, a heart-on-a-chip platform, incorporating microgrooves and electrical pulse stimulations to recapitulate the well-aligned structure and synchronous beating of cardiomyocytes (CMs) for drug screening, is reported. Each chip is made with facile lithographic and laser-cutting processes that can be easily scaled up to high-throughput format. The maturation and phenotypic changes of CMs cultured on the heart-on-a-chip is validated and it can be treated with various drugs to evaluate cardiotoxicity and cardioprotective efficacy. The heart-on-a-chip can provide a high-throughput drug screening platform in preclinical drug development.


Differences between sexually abused and non-sexually abused adolescent girls in foster care.
This descriptive study examines the differences between sexually abused and non-sexually abused adolescent females in the foster care system who were participating in an independent living program. Fifty-four percent of the 190 girls met the criteria for being categorized as sexually abused. Those who experienced sexual abuse had also experienced significantly more of other types of child maltreatment. In addition, those who had been sexually abused were much more likely to be living in a congregate living setting, such as a group home or residential center, than those who were not sexually abused. The girls who had been sexually abused exhibited significantly more behavioral difficulties, including internalizing and externalizing problems, with 51% of them having clinically significant scores on the Youth Self-Report version of the Child Behavior Checklist. When co-occurrence of substance use and mental health problems were examined, sexually abused girls were significantly more likely than the non-sexually abused girls to meet the established criteria.


[Ambulatory short-term therapy of anxiety patients with autogenic training and hypnosis. Results of treatment and 3 months follow-up].
The aim of the study was to examine the effects of a short-term outpatient treatment (6 sessions à 90 minutes) with autogenic training (Schultz 1932) or hypnosis (Erickson and Rossi 1979), concerning patients with anxiety- and panic disorders (according to DSM-III-R and ICD-10-criteria). 27 Outpatients were recruited by newspaper articles. Most of them had not been treated before. Both short-term treatments consisted of 6 weekly group sessions (4-6 patients). Psychological examinations were carried out before and after treatment and at a follow-up after 3 months. Trait anxiety (STAI-X2) and psychosomatic complaints (BfS/BL') decreased in both groups significantly after treatment. This therapeutic effect was stable during the following 3 months. The frequency of panic attacks decreased as well under treatment, and this effect was even more marked during the 3 month following treatment. The therapeutic success was influenced by the frequency of autosuggestive exercises carried out by the patients at home. This form of short-term treatment is not sufficient for all patients with anxiety disorders but the results of that study encourage to use short-term interventions with autogenic training and hypnosis as initial therapeutic approaches for such patients and to combine them with other therapeutic interventions.


[Clinical significance of the human epidermal growth factor receptor 2( HER2) in patients with recurrent gastric cancer who received S-1 adjuvant chemotherapy].
The aim of this study was to clarify the human epidermal growth factor receptor 2( HER2) positivity, clinicopathological characteristics, and survival of patients with recurrent HER2-positive gastric cancer who received S-1 adjuvant chemotherapy. Thirty-eight patients with recurrent gastric cancer who underwent curative D2 surgery and received S-1 adjuvant chemotherapy between June 2002 and December 2011 were examined. HER2 positivity was determined as defined in the ToGA study. The positivity score was assessed by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) as follows: IHC 0 in 27 patients, IHC 1+in 4, IHC 2+/FISH-in 3, IHC 2+/FISH+in 1, and IHC 3+in 3. The HER2 positivity rate was 10.5% (4/38). HER2-positive recurrent gastric cancer was characterized by a differentiated histological feature and frequent blood vessel invasion. However, the recurrence and survival rates were not significantly different between the HER2-negative and HER2-positive tumors. The HER2 positivity rate after S-1 adjuvant chemotherapy did not differ significantly between patients with recurrent gastric cancer and those with primary Stage II/III gastric cancer, suggesting that S-1 adjuvant chemotherapy was equally effective, regardless of HER2 status.


Role of diet and gut microbiota on colorectal cancer immunomodulation.
Colorectal cancer (CRC) is one of the most commonly diagnosed cancers, and it is characterized by genetic and epigenetic alterations, as well as by inflammatory cell infiltration among malignant and stromal cells. However, this dynamic infiltration can be influenced by the microenvironment to promote tumor proliferation, survival and metastasis or cancer inhibition. In particular, the cancer microenvironment metabolites can regulate the inflammatory cells to induce a chronic inflammatory response that can be a predisposing condition for CRC retention. In addition, some nutritional components might contribute to a chronic inflammatory condition by regulating various immune and inflammatory pathways. Besides that, diet strongly modulates the gut microbiota composition, which has a key role in maintaining gut homeostasis and is associated with the modulation of host inflammatory and immune responses. Therefore, diet has a fundamental role in CRC initiation, progression and prevention. In particular, functional foods such as probiotics, prebiotics and symbiotics can have a potentially positive effect on health beyond basic nutrition and have anti-inflammatory effects. In this review, we discuss the influence of diet on gut microbiota composition, focusing on its role on gut inflammation and immunity. Finally, we describe the potential benefits of using probiotics and prebiotics to modulate the host inflammatory response, as well as its application in CRC prevention and treatment.


Metformin Suppresses Tumor Progression by Inactivating Stromal Fibroblasts in Ovarian Cancer.
Ovarian cancer is a devastating disease due to its high incidence of relapse and chemoresistance. The tumor microenvironment, especially the tumor stroma compartment, was proven to contribute tremendously to the unsatisfactory chemotherapeutic efficacy in ovarian cancer. Cytotoxic agents not only effect tumor cells, but also modulate the phenotype and characteristics of the vast stromal cell population, which can in turn alter the tumor cell response to chemointervention. In this study, we focused on the tumor stroma response to cytotoxic agents and the subsequent effect on the ovarian cancer tumor cells. First, we found a significant stromal overexpression of IL6 in patient samples that received cisplatin-based treatment, which was further validated in purified fibroblasts challenged with cisplatin. Stromal fibroblast-derived IL6 was proven to mediate ovarian cancer tumor cell chemoresistance. For the first time, we found that the tumor stroma of patients with routine metformin administration exhibited lower IL6 expression. Thus, we presumed that metformin was a potent alleviator of stromal inflammation in ovarian cancer. We found that metformin partly reversed cisplatin-stimulated IL6 secretion in the stromal fibroblasts and attenuated fibroblast-facilitated tumor growth in 3D organotypic cocultures and murine xenograft models. Mechanistically, we found that metformin inhibited IL6 secretion via suppressing NFκB signaling, an upstream controller of stromal inflammation. Collectively, our findings introduced a novel mechanism of metformin in suppressing ovarian cancer progression through diminishing chemotherapy-induced stromal activation. Therefore, we provide an alternative therapeutic option in targeting stromal inflammation and a potential scheme of combination therapy to improve the chemosensitivity in ovarian cancer. Mol Cancer Ther; 17(6); 1291-302. ©2018 AACR.


From rigid bone plate fixation to stable dynamic osteosynthesis in mandibular and craniomaxillo-facial surgery: Historical evolution of concepts and technical developments.
Historically, extensive observation of limb fracture healing led to a consensus that only complete rigid immobilization could guarantee recovery. This agreement was arrived at because for a long time progress in treatment was driven by clinicians and did not stem from the application of biological research. The clinical approach was based on immobilization of the fracture by rigid osteosynthesis plates and bicortical screws. Subsequently, after extrapolation of the ideas of Lane, the concept of rigid compressive osteosynthesis rapidly gained in acceptance. It was not until the second half of the 20th century that maxillofacial surgeons concluded that the principles of osteosynthesis should be based on biomechanical studies and not only on clinical observation. The concept of stable dynamic osteosynthesis stems from basic research. This paper traces the evolution of concepts in maxillofacial osteosynthesis.


Thioredoxin reductase is essential for the survival of Plasmodium falciparum erythrocytic stages.
The human malaria parasite Plasmodium falciparum poses an increasing threat to human health in the tropical regions of the world, and the validation and assessment of possible drug targets is required for the development of new antimalarials. It has been shown that the erythrocytic stages of the parasites, which are responsible for the pathology of the disease in humans, are under enhanced oxidative stress and are particularly vulnerable to exogenous challenges by reactive oxygen species. Therefore it is postulated that the disruption of the antioxidant and/or redox systems of the parasite is a feasible way to interfere with their development during erythrocytic schizogony. In order to test this suggestion thioredoxin reductase (TrxR), an enzyme heavily involved in maintenance of redox homeostasis and antioxidant defense, was knocked out in P. falciparum. It was impossible to generate parasites with a disrupted trxR gene suggesting that TrxR is essential for P. falciparum erythrocytic stages. Technical problems were excluded by transfecting a 3' replacement construct, which recombined correctly and transfectants did not show any phenotypic alterations. In order to prove that the trxR knockout was responsible for the lethal phenotype of the null mutants, a co-transfection with both the knockout construct and a construct containing the trxR coding region under the control of the calmodulin promoter was conducted. Despite the disruption of the trxR gene, parasites were viable. In a Southern blot analysis a complicated restriction pattern was obtained, but it was shown by pulse field gel electrophoresis and field inverse gel electrophoreses that only the trxR gene locus on chromosome 9 was targeted by the constructs. It was found that the co-transfected constructs form concatemeric structures prior to integration into the trxR gene locus, which is further supported by plasmid rescue followed by restriction analyses of the plasmids. Northern and Western blot analyses proved that the co-transfectants highly overexpress TrxR from the introduced gene. Our results demonstrate that TrxR is essential for the survival of the erythrocytic stages of P. falciparum.


Syntactic breakdown and recovery of clausal structure in agrammatism.
We studied the recovery of clause structures in four agrammatic patients by performing a longitudinal analysis of their spontaneous production. We classified their utterances in terms of legitimacy of the syntactic structure and rate of subordination. The results show that, initially, patients omit verbs, avoid subordination, just employ present tense, and substitute finite verbs with infinitives. During the recovery they start employing other tenses and subordinate clauses and reduce the use of infinitives. Data suggest a mixed syntactic and morphological origin of the impairment. Assuming that phrasal representations are built bottom-up from an array of lexical items, we propose that syntactic structures recover stepwise, with the lower portion of the syntactic tree becoming accessible before higher portions. We claim that, despite superficial similarities between agrammatic and children's speech, e.g., use of infinitives instead of finite verbs, a unified account is not viable.


Pharmacology of selective thrombin inhibitors.
Thrombin plays a key role in thrombosis and haemostasis, and is selectively inhibited by hirudin and synthetic inhibitors. Hirudin, a polypeptide (molecular weight 7,000 daltons) extracted from medicinal leeches, can now be produced by gene technology. Hirudin binds selectively to thrombin with high affinity and inhibits its enzymatic properties. Besides heparin, hirudin is not inhibited by platelet factor 4; it prolongs in vitro and ex vivo routine blood coagulation assays and prevents thrombosis in a number of animal models without increasing haemorrhagic risk. In humans, hirudin disappears from the blood with a half-life of 1 h, is devoid of undesirable side effects and has been shown to be efficient in the treatment of chronic disseminated intravascular coagulation (DIC). A number of synthetic direct thrombin inhibitors have been described, including benzamidine derivatives which share identical pharmacological properties with hirudin; however their biological half-life after i.v. injection is shorter. Other derivatives (amidino-phenyl-pyruvic acid) have longer half-lives and have been used to treat chronic DIC in man.


Serotoninergic and melatoninergic systems are fully expressed in human skin.
We investigated the cutaneous expression of genes and enzymes responsible for the multistep conversion of tryptophan to serotonin and further to melatonin. Samples tested were human skin, normal and pathologic (basal cell carcinoma and melanoma), cultured normal epidermal and follicular melanocytes, melanoma cell lines, normal neonatal and adult epidermal and follicular keratinocytes, squamous cell carcinoma cells, and fibroblasts from dermis and follicular papilla. The majority of the samples showed simultaneous expression of the genes for tryptophan hydroxylase, arylalkylamine N-acetyltransferase (AANAT), and hydroxyindole-O-methyltransferase (HIOMT). The products of AANAT activity were identified by RP-HPLC with fluorimetric detection in human skin and in cultured normal and malignant melanocytes and immortalized keratinocytes; HIOMT activity was detected in human skin, keratinocytes, and melanoma cells. N-acetylserotonin (NAS) was detected by RP-HPLC in human skin extracts. NAS identity was confirmed further by LC/MS in keratinocytes. In conclusion, we provide evidence that the human skin expresses intrinsic serotonin and melatonin biosynthetic pathways.


Acrylfentanyl: Another new psychoactive drug with fatal consequences.
The European Nordic Countries are the most exposed to opioid-related deaths. Between April and October 2016, a series of forty lethal intoxications occurred in Sweden, in which the presence of the synthetic opioid acrylfentanyl was determined to be the main - or a contributing - cause of death. In the reported cases, the blood concentration of acrylfentanyl - mostly detected in combination with other drugs - ranged from 0.01ng/g to 5ng/g; victims were predominantly males (34 males and 6 females), and their age varied between 18 and 53 years. We further describe five cases, representative of the different drug administration route (nasal spray, tablets) and intentions (accidental or voluntary intoxication). Moreover, we address nine cases of non-lethal intoxication, in single (8 cases) or polydrug scenario (1 case). We discuss the present characteristics of the Swedish drug market for fentanyl-analogs in general and acrylfentanyl in particular, reporting a structural difficulty to effectively counteracting the appearance of unscheduled substances due to the constant turnover of new molecules on the recreational drug market.