Liver transplantation for Wilson's disease: a single-center experience.
Wilson's disease is a hereditary defect in copper excretion leading to the accumulation of copper in the tissues, with subsequent tissue damage. The most serious sequela is that of progressive central nervous system involvement. The use of orthotopic liver transplantation (OLT) has been controversial for those patients with neurological symptoms attributed to Wilson's disease. The aim of this study is to determine the effectiveness of OLT for patients with Wilson's disease, including those with neurological involvement attributed to copper accumulation in the central nervous system. OLT was performed in 45 patients (19 men [42.2%], 26 women [57.8%]) with Wilson's disease between 1971 and 1993 who were followed up for at least 4 years. The age at diagnosis of Wilson's disease ranged from 3 to 41 years (mean, 17.7 +/- 7.4 years). The age at OLT ranged from 8 to 52 years (mean, 22.3 +/- 9.4 years). Nineteen patients (42.2%) were aged younger than 18 years at OLT. The indications for OLT included chronic hepatic failure in 15 patients (33.3%) and fulminant (FHF) or subfulminant hepatic failure in 30 patients (66. 6%). All but 1 of the 19 pediatric patients (94.7%) were in the latter group. Twenty-five patients (55.5%) were receiving D-penicillamine, 9 patients for more than 1 year; none of the patients treated long term presented as FHF. Thirty-three patients (73.3%) survived more than 5 years after OLT. Fourteen patients (31%) died during the posttransplantation period; 7 of the 14 patients (50%) were aged younger than 18 years. Twelve patients died during the first 3 months after OLT of complications of disease and surgery, 10 of whom underwent transplantation for FHF. The other 2 patients died 6 and 9 years after transplantation of infectious problems. Eleven patients (24.4%) required retransplantation because of a primary nonfunctioning graft (n = 6), chronic rejection (n = 4), and hepatic artery thrombosis (n = 1). Seventeen patients (37.7%) presented with neurological abnormalities; 14 patients with Wilsonian neurological manifestations and 3 patients with components of increased intracranial pressure. Ten of the 13 surviving patients with hepatic insufficiency and neurological abnormalities at OLT showed significant neurological improvement. Our experience shows OLT is a life-saving procedure in patients with end-stage Wilson's disease and is associated with excellent long-term survival. The neurological manifestation of the disease can improve significantly after OLT. Earlier transplantation in patients with an unsatisfactory response to medical treatment may prevent irreversible neurological deterioration and less satisfactory improvement after OLT.


Novel nuclear and cytoplasmic proteins detected by anti-Zoothamnium arbuscula (Protozoa) spasmin 1 antibody in mammalian cells are dependent on the cell cycle.
Spasmin is a calcium-binding protein that is the major component of calcium-induced contractile filaments, called spasmoneme, found in vorticellid ciliates. Such filaments have not been observed in any organisms other than green algae. To determine whether calcium-induced contractile filaments resembling spasmoneme are present in higher eukaryotes, we performed immunofluorescence imaging of an anti-Zoothamnium arbuscula (protozoa, ciliophora) spasmin 1 polyclonal antibody in HeLa cells. In the cytoplasm, ubiquitous antigens seemed to be co-localized with microtubules at interphase, but not throughout mitosis. In the nucleus, areas linked to the nuclear envelope contained a number of hot spots. These regions were unclear during condensation of the replicated chromosomes, but became clearly visible again at cytokinesis. Immunoblotting analysis identified localized antigens during different phases of the cell cycle, including a 68/71 kDa cytoplasmic protein and a 55 kDa nuclear protein in interphase, and a 55/70 kDa protein in mitosis. The anti-spasmin 1 antibody recognized antigens in both hamster kidney BHK21 cells and Human lung cancer A-549 cells. These results suggest that novel spasmin-like proteins could be common in mammalian cells.


Characterization and simulation of electrolyte-gated organic field-effect transistors.
In this work we fabricate and characterize field-effect transistors based on the solution-processable semiconducting polymer poly(3-hexylthiophene) (P3HT). Applying two independent gate potentials to the electrolyte-gated organic field-effect transistor (EGOFET), by using a conventional SiO(2) layer as the back-gate dielectric and the electrolyte-gate as the top-gate, allows the measurement of the electrical double layer (EDL) capacitance at the semiconductor-electrolyte interface. We record the transfer curves of the transistor in salt solutions of different concentration by sweeping the bottom gate potential for various constant electrolyte-gate potentials. A change of the electrolyte-gate potential towards more negative voltages shifts the threshold voltage of the bottom-gate channel towards more positive back-gate potentials, which is directly proportional to the capacitive coupling factor. By operating the EGOFET in the dual-gate mode, we can prove the dependency of the EDL capacitance on the molarity of the electrolyte according to the Debye-Hückel theory, and additionally show the difference between a polarizable and non-polarizable electrolyte-gate electrode. With the experimentally obtained values for the EDL capacitance at the semiconductor-electrolyte interface we can model the electrolyte-gate transfer characteristics of the P3HT OTFT.


Yeast Rad7-Rad16 complex, specific for the nucleotide excision repair of the nontranscribed DNA strand, is an ATP-dependent DNA damage sensor.
In eukaryotes, nucleotide excision repair of ultraviolet light-damaged DNA is a highly intricate process that requires a large number of evolutionarily conserved protein factors. Genetic studies in the yeast Saccharomyces cerevisiae have indicated a specific role of the RAD7 and RAD16 genes in the repair of transcriptionally inactive DNA. Here we show that the RAD7- and RAD16-encoded products exist as a complex of 1:1 stoichiometry, exhibiting an apparent dissociation constant (Kd) of <4 x 10(-10) M. The Rad7-Rad16 complex has been purified to near homogeneity in this study and is shown to bind, in an ATP-dependent manner and with high specificity, to DNA damaged by ultraviolet light. Importantly, inclusion of the Rad7-Rad16 complex in the in vitro nucleotide excision repair system that consists entirely of purified components results in a marked stimulation of damage specific incision. Thus, Rad7-Rad16 complex is the ATP-dependent DNA damage sensor that specifically functions with the ensemble of nucleotide excision repair factor (NEF) 1, NEF2, NEF3, and replication protein A in the repair of transcriptionally inactive DNA. We name this novel complex of Rad7 and Rad16 proteins NEF4.


Control of Listeria monocytogenes in ready-to-eat meats containing sodium levulinate, sodium lactate, or a combination of sodium lactate and sodium diacetate.
This study investigated the use of sodium levulinate to prevent outgrowth of Listeria monocytogenes in refrigerated ready-to-eat (RTE) meat products. Turkey breast roll and bologna were formulated to contain 1%, 2%, or 3% (w/w) sodium levulinate, 2% sodium lactate, a 2% combination of sodium lactate and sodium diacetate (1.875% sodium lactate and 0.125% sodium diacetate), or no antimicrobial (control). Samples of the RTE products were sliced, inoculated with 10(2) to 10(3) CFU/cm(2) of a 5-strain cocktail of L. monocytogenes, vacuum packaged, and stored at refrigeration temperature for 0 to 12 wk. Counts reached 10(8) CFU/cm(2) on control turkey roll product after 8 wk, and over 10(7) CFU/cm(2) on control bologna after 12 wk. Addition of 2% or more sodium levulinate to turkey roll and 1% or more sodium levulinate to bologna completely prevented growth of L. monocytogenes during 12 wk of refrigerated storage. A consumer taste panel with pathogen-free samples found no differences in the overall liking among the preparations of turkey roll or among preparations of bologna. These results show that sodium levulinate is at least as effective at inhibiting outgrowth of L. monocytogenes in RTE meat products as the current industry standards of lactate or lactate and diacetate, and levulinate addition does not alter the overall liking of the RTE meat products.


Tumor size in endometrial cancer.
Tumor size was determined in 142 patients with clinical Stage I endometrial cancer treated primarily by total abdominal hysterectomy, bilateral salpingo-oophorectomy, and lymph node biopsies between July 1979 and August 1988. Only 4% of patients with tumor size less than or equal to 2 cm had lymph node metastasis; this increased to 15% for tumors more than 2 cm and increased further to 35% when the entire uterine cavity was involved (multivariate P = 0.01). Five-year survival was 98% for patients with tumors less than or equal to 2 cm, 84% with tumors more than 2 cm, and 64% with tumors involving the whole uterine cavity (Mantel-Cox P = 0.005). For endometrial cancer patients with Grade 2 tumors and less than one-half myometrial invasion, the risk of lymph node metastasis is often considered too low to justify adjuvant pelvic radiation therapy. This intermediate-risk group is better defined by including tumor size as a prognostic factor. For this subgroup (Grade 2, less than one-half endometrial invasion) there were no lymph node metastasis associated with tumors less than 2 cm, but 18% had nodal disease when tumors were larger than 2 cm. Tumor size is an important prognostic factor that is particularly helpful in directing adjuvant radiation therapy in patients without staging lymph node biopsies.


The hereditary adult-onset ataxias in South Africa.
There is little data on the spectrum and frequencies of the autosomal dominant spinocerebellar ataxias (SCAs) from the African continent. We undertook a large prospective population-based study over a 10-year period in South Africa (SA). Affected persons were clinically evaluated, and the molecular analysis for the SCA1, 2, 3, 6 and 7 expansions was undertaken. Of the 54 SA families with dominant ataxia, SCA1 accounted for 40.7%, SCA2 for 13%, SCA3 for 3.7%, SCA6 for 1.9%, SCA7 for 22.2% and 18.5% were negative for all these mutations. The frequency of the SCA1 and SCA7 expansions in SA represents one of the highest frequencies for these expansions reported in any country. In this study, the SCA7 mutations have only been found in SA families of Black ethnic origin.


Argonaute CLIP-Seq reveals miRNA targetome diversity across tissue types.
To date, analyses of individual targets have provided evidence of a miRNA targetome that extends beyond the boundaries of messenger RNAs (mRNAs) and can involve non-Watson-Crick base pairing in the miRNA seed region. Here we report our findings from analyzing 34 Argonaute HITS-CLIP datasets from several human and mouse cell types. Investigation of the architectural (i.e. bulge vs. contiguous pairs) and sequence (Watson-Crick vs. G:U pairs) preferences for human and mouse miRNAs revealed that many heteroduplexes are "non-canonical" i.e. their seed region comprises G:U and bulge combinations. The genomic distribution of miRNA targets differed distinctly across cell types but remained congruent across biological replicates of the same cell type. For some cell types intergenic and intronic targets were more frequent whereas in other cell types mRNA targets prevailed. The findings suggest an expanded model of miRNA targeting that is more frequent than the standard model currently in use. Lastly, our analyses of data from different cell types and laboratories revealed consistent Ago-loaded miRNA profiles across replicates whereas, unexpectedly, the Ago-loaded targets exhibited a much more dynamic behavior across biological replicates.


Melanopsin contributions to non-visual and visual function.
Melanopsin is a short-wavelength-sensitive photopigment that was discovered only around 20 years ago. It is expressed in the cell bodies and processes of a subset of retinal ganglion cells in the retina (the intrinsically photosensitive retinal ganglion cells; ipRGCs), thereby allowing them to signal light even in the absence of cone and rod input. Many of the fundamental properties of melanopsin signalling in humans for both visual (e.g. detection, discrimination, brightness estimation) and non-visual function (e.g. melatonin suppression, circadian phase shifting) remain to be elucidated. Here, we give an overview of what we know about melanopsin contributions in visual function and non-visual function.


Role of newer and re-emerging older agents in the treatment of infections caused by carbapenem-resistant Enterobacteriaceae.
Antimicrobial resistance has been identified by the World Health Organization as "one of the three greatest threats to human health." Gram negative bacteria in particular drive this alarming trend. Carbapenem-resistant Enterobacteriaceae (CRE) such as Escherichia coli, Klebsiella pneumoniae, and Enterobacter species are of particular importance as they are associated with poor clinical outcomes and are common causes for a variety of infections including bacteremia, urinary tract infection, intra-abdominal infections and pneumonia. CRE are difficult to treat as carbapenem resistance is often accompanied by resistance to additional drug classes. For example, CRE may be extensively drug resistant or even pandrug resistant. Unfortunately, CRE infections have increased over the past 15 y while new and effective antibiotics have not kept pace. Recently, however, new agents have become available to help treat CRE infection, and several more are under development. This article reviews the efficacy, safety, and pharmacokinetic issues around 4 emerging agents to treat CRE - ceftazidime-avibactam, fosfomycin, tigecycline, and minocycline. In addition, an overview of agents in the antibiotic pipeline - meropenem-vaborbactam, imipenem-relebactam, plazomicin, and eravacycline is provided. More established agents, such as those in the polymyxin class and aminoglycoside class (other than the pipeline agent plazomicin), are not addressed here.


Atrophy rates in asymptomatic amyloidosis: implications for Alzheimer prevention trials.
There is considerable interest in designing therapeutic studies of individuals at risk of Alzheimer disease (AD) to prevent the onset of symptoms. Cortical β-amyloid plaques, the first stage of AD pathology, can be detected in vivo using positron emission tomography (PET), and several studies have shown that ~1/3 of healthy elderly have significant β-amyloid deposition. Here we assessed whether asymptomatic amyloid-PET-positive controls have increased rates of brain atrophy, which could be harnessed as an outcome measure for AD prevention trials. We assessed 66 control subjects (age = 73.5±7.3 yrs; MMSE = 29±1.3) from the Australian Imaging Biomarkers & Lifestyle study who had a baseline Pittsburgh Compound B (PiB) PET scan and two 3T MRI scans ~18-months apart. We calculated PET standard uptake value ratios (SUVR), and classified individuals as amyloid-positive/negative. Baseline and 18-month MRI scans were registered, and brain, hippocampal, and ventricular volumes and annualized volume changes calculated. Increasing baseline PiB-PET measures of β-amyloid load correlated with hippocampal atrophy rate independent of age (p = 0.014). Twenty-two (1/3) were PiB-positive (SUVR>1.40), the remaining 44 PiB-negative (SUVR≤1.31). Compared to PiB-negatives, PiB-positive individuals were older (76.8±7.5 vs. 71.7±7.5, p<0.05) and more were APOE4 positive (63.6% vs. 19.2%, p<0.01) but there were no differences in baseline brain, ventricle or hippocampal volumes, either with or without correction for total intracranial volume, once age and gender were accounted for. The PiB-positive group had greater total hippocampal loss (0.06±0.08 vs. 0.02±0.05 ml/yr, p = 0.02), independent of age and gender, with non-significantly higher rates of whole brain (7.1±9.4 vs. 4.7±5.5 ml/yr) and ventricular (2.0±3.0 vs. 1.1±1.0 ml/yr) change. Based on the observed effect size, recruiting 384 (95%CI 195-1080) amyloid-positive subjects/arm will provide 80% power to detect 25% absolute slowing of hippocampal atrophy rate in an 18-month treatment trial. We conclude that hippocampal atrophy may be a feasible outcome measure for secondary prevention studies in asymptomatic amyloidosis.


Nutrient intake from food in children with autism.
The impact of abnormal feeding behaviors reported for children with autism spectrum disorders (ASDs) on their nutritional status is unknown. We compared nutrient intake from food consumed by children with and without ASD and examined nutrient deficiency and excess. Prospective 3-day food records and BMI for children (2-11 years) with ASD participating in the Autism Treatment Network (Arkansas, Cincinnati, Colorado, Pittsburgh, and Rochester) were compared with both the National Health and Nutrition Examination Survey data and a matched subset based on age, gender, family income, and race/ethnicity (N = 252 analyzed food records). Children with ASD and matched controls consumed similar amounts of nutrients from food. Only children with ASD aged 4 to 8 years consumed significantly less energy, vitamins A and C, and the mineral Zn; and those 9 to 11 years consumed less phosphorous. A greater percentage of children with ASD met recommendations for vitamins K and E. Few children in either group met the recommended intakes for fiber, choline, calcium, vitamin D, vitamin K, and potassium. Specific age groups consumed excessive amounts of sodium, folate, manganese, zinc, vitamin A (retinol), selenium, and copper. No differences were observed in nutritional sufficiency of children given restricted diets. Children aged 2 to 5 years with ASD had more overweight and obesity, and children 5 to 11 years had more underweight. Children with ASD, like other children in America, consume less than the recommended amounts of certain nutrients from food. Primary care for all children should include nutritional surveillance and attention to BMI.


Altered histology of the thymus and spleen in contaminant-exposed juvenile American alligators.
Morphological differences in spleen and thymus are closely related to functional immune differences. Hormonal regulation of the immune system has been demonstrated in reptilian splenic and thymic tissue. Spleens and thymus were obtained from juvenile alligators at two reference sites in Florida, USA: Orange Lake and a National Wildlife Refuge, Lake Woodruff, as well as from a contaminated lake, Lake Apopka. Lake Apopka has been extensively polluted with agricultural pesticides. Tissues were prepared for histological analysis to determine if previously detected endocrine abnormalities associated with contaminant exposure might also be reflected in morphological differences in splenic and thymic structures important for immunological response. Similar tissues were taken from captive-raised juvenile female alligators (3 years old) that were hatched from eggs collected on Lake Woodruff and Lake Apopka. Differences in thymic ratios (medulla/cortex) were found among alligators collected from the two lakes (P = 0.0051). Alligators from Lake Apopka had smaller thymic ratios than animals from either reference lake. Males from Lake Woodruff had significantly smaller lymphocyte sheaths in the spleen than females (P = 0.0009), indicative of a normal sexual dimorphism. Lymphocyte sheath width differed among females obtained from the three lakes, with females from Lake Apopka having the smallest sheath width and those from Orange Lake having the largest. Malpighian body area was largest in alligators from Orange Lake, intermediate in Lake Woodruff, and smallest in Lake Apopka. In contrast to that observed for wild-caught animals, no difference was found in the thymic medulla/cortex ratio of captive-raised female alligators (P = 0.378). Captive-raised female alligators from Lake Apopka and Lake Woodruff displayed lake-associated differences in lymphocyte sheath width as observed in wild animals; Lake Apopka alligators had smaller lymphocyte sheath width compared to Woodruff alligators (P = 0.0396). In contrast to wild-caught animals, area of the Malpighian bodies did not differ by lake in the captive-raised female alligators (P = 0.066). The enlarged thymic cortex suggests a change in T-lymphocyte maturation within the thymus of alligators from a contaminated environment, Lake Apopka. The results point to alterations in the histology of the thymus and spleen. Further studies are required to examine the functional significance of these observations.


γ-(S)-Trifluoromethyl proline: evaluation as a structural substitute of proline for solid state (19)F-NMR peptide studies.
γ-(4S)-Trifluoromethyl proline was synthesised according to a modified literature protocol with improved yield on a multigram scale. Conformational properties of the amide bond formed by the amino acid were characterised using N-acetyl methyl ester model. The amide populations (s-trans vs. s-cis) and thermodynamic parameters of the isomerization were found to be similar to the corresponding values for intact proline. Therefore, the γ-trifluoromethyl proline was suggested as a structurally low-disturbing proline substitution in peptides for their structural studies by (19)F-NMR. Indeed, the exchange of native proline for γ-trifluoromethyl proline in the peptide antibiotic gramicidin S was shown to preserve the overall amphipathic peptide structure. The utility of the amino acid as a selective (19)F-NMR label was demonstrated by observing the re-alignment of the labelled gramicidin S in oriented lipid bilayers.


Association Between Statin Use and Endometrial Cancer Survival.
To evaluate the association of 3 hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibitor (statin) use and concordant polypharmacy with disease-specific survival from endometrial cancer. A retrospective cohort study was conducted of 985 endometrial cancer cases treated from January 1999 through December 2009 at a single institution. Disease-specific survival was estimated by Kaplan-Meier analyses. A Cox proportional hazards model was used to study factors associated with survival. All statistical tests were two-sided and performed using Stata. At the time of analysis, 230 patients (22% of evaluable patients) died of disease and median follow-up was 3.28 years. Disease-specific survival was greater (179/220 [81%]) for women with endometrial cancer taking statin therapy at the time of diagnosis and staging compared with women not using statins (423/570 [74%]) (log rank test, P=.03). This association persisted for the subgroup of patients with nonendometrioid endometrial tumors who were statin users (59/87 [68%]) compared with nonusers (93/193 [43%]) (log rank test, P=.02). The relationship remained significant (hazard ratio 0.63, 95% confidence interval [CI] 0.40-0.99) after adjusting for age, clinical stage, radiation, and other factors. Further evaluation of polypharmacy showed an association between concurrent statin and aspirin use with an especially low disease-specific mortality (hazard ratio 0.25, 95% CI 0.09-0.70) relative to those who used neither. Statin and aspirin use was associated with improved survival from nonendometrioid endometrial cancer.


The Brief International Cognitive Assessment in Multiple Sclerosis (BICAMS): results from the German validation study.
Recent research has convincingly shown that the ability to work mainly depends on the cognitive status in multiple sclerosis (MS). An international committee of experts recommended a brief neuropsychological battery to evaluate cognitive performance in MS. BICAMS comprises three tests, the Symbol Digit Modalities Test (SDMT), the learning trials of the California Verbal Learning Test II (CVLT-II), and the Brief Visuospatial Memory Test-Revised (BVMT-R). To validate BICAMS on a sample of German MS patients and healthy controls (HCs). According to the international guidelines for validation, examiner's instructions were standardized and translated into German. Due to the availability of better normative data for future applications in routine clinical care and classification of individual performance degree, the Rey Auditory Verbal Learning Test (RAVLT) (German version: Verbaler Lern- und Merkfähigkeits-Test, VLMT) was chosen instead of CVLT-II. 172 MS patients and 100 HCs entered the study. BICAMS was administered at baseline and retest (after 3-4 weeks). The groups did not differ in age, gender or education. Mean age of MS patients was 43.33 years (SD 11.64); 68% were female and 86.9% had relapsing-remitting MS. Patients performed significantly worse than HCs on the SDMT (p < 0.01) and on BVMT-R (p < 0.05) but not on VLMT. In addition, BICAMS was shown to be reliable over time: r = 0.71 for BVMT-R, r = 0.72 for VLMT and r = 0.85 for SDMT. SDMT z-score proved to be a good predictor for the ability to work in a full-time (p < 0.001) as well as in a part-time job (p < 0.001). VLMT z-score turned out to be a significant predictor only for the ability to work in a part-time job, while BVMT-R z-score showed no significant predictive value. In this German validation study with the VLMT, the modified BICAMS (BICAMS-M) turned out to reliably detect cognitive problems in MS patients and to monitor cognitive performance over time. SDMT revealed the best predictive value for working ability. Moreover, only the SDMT was able to predict the ability to work in a part-time or full-time job. Following these results, application of the SDMT is recommended for medical statements on working ability of MS patients.


Signal-, set-, and movement-related activity in the human premotor cortex.
Single unit recording studies in non-human premotor cortex have revealed neurons with motor-related activity. Other neurons, however, seem to be involved in prior movement selection and preparation processes, and have activity related to visual instruction signals or movement preparation ('set'). We have used single pulse transcranial magnetic stimulation (TMS) to identify similar processes in human subjects. In Experiment 1 subjects performed a cued movement task while being stimulated with TMS over three sites: sensorimotor cortex, posterior premotor cortex and anterior premotor cortex. TMS slowed movements when applied at 140 ms after the visual cue over the anterior premotor site, at 180 ms after the visual cue over the posterior premotor site, and at 220 ms and later after the visual cue over the sensorimotor cortex. The results are consistent with a change from signal to movement-related processing when moving from premotor to motor cortex. In Experiment 2 there was a preparatory set period between the instruction signal that informed subjects which movement to make and the 'go' signal that informed them when to actually make the movement. TMS was applied over the anterior premotor site and the sensorimotor site during the set period. At both sites TMS had similar effects on slowing subsequent movements. The results suggest set activity in both premotor and motor cortices in human subjects.


Diagnostic potential of sarcoplasmic myxovirus resistance protein A expression in subsets of dermatomyositis.
To elucidate the diagnostic value of sarcoplasmic expression of myxovirus resistance protein A (MxA) for dermatomyositis (DM) specifically analysing different DM subforms, and to test the superiority of MxA to other markers. Immunohistochemistry for MxA and retinoic acid-inducible gene I (RIG-I) was performed on skeletal muscle samples and compared with the item presence of perifascicular atrophy (PFA) in 57 DM patients with anti-Mi-2 (n = 6), -transcription intermediary factor 1 gamma (n = 10), -nuclear matrix protein 2 (n = 13), -melanoma differentiation-associated gene 5 (MDA5) (n = 10) or -small ubiquitin-like modifier activating enzyme (n = 1) autoantibodies and with no detectable autoantibody (n = 17). Among the patients, nine suffered from cancer and 22 were juvenile-onset type. Disease controls included antisynthetase syndrome (ASS)-associated myositis (n = 30), immune-mediated necrotizing myopathy (n = 9) and inclusion body myositis (n = 5). Sarcoplasmic MxA expression featured 77% sensitivity and 100% specificity for overall DM patients, while RIG-I staining and PFA reached respectively 14% and 59% sensitivity and 100% and 86% specificity. In any subset of DM, sarcoplasmic MxA expression showed higher sensitivity than RIG-I and PFA. Some anti-MDA5 antibody-positive DM samples distinctively showed a scattered staining pattern of MxA. No ASS samples had sarcoplasmic MxA expression even though six patients had DM skin rash. Sarcoplasmic MxA expression is more sensitive than PFA and RIG-I expression for a pathological diagnosis of DM, regardless of the autoantibody-related subgroup. In light of its high sensitivity and specificity, it may be considered a pathological hallmark of DM per se. Also, lack of MxA expression in ASS supports the idea that ASS is a distinct entity from DM.


DNA Microarray for Rapid Detection and Identification of Food and Water Borne Bacteria: From Dry to Wet Lab.
A rapid, accurate, flexible and reliable diagnostic method may significantly decrease the costs of diagnosis and treatment. Designing an appropriate microarray chip reduces noises and probable biases in the final result. The aim of this study was to design and construct a DNA Microarray Chip for a rapid detection and identification of 10 important bacterial agents. In the present survey, 10 unique genomic regions relating to 10 pathogenic bacterial agents including Escherichia coli (E.coli), Shigella boydii, Sh.dysenteriae, Sh.flexneri, Sh.sonnei, Salmonella typhi, S.typhimurium, Brucella sp., Legionella pneumophila, and Vibrio cholera were selected for designing specific long oligo microarray probes. For this reason, the in-silico operations including utilization of the NCBI RefSeq database, Servers of PanSeq and Gview, AlleleID 7.7 and Oligo Analyzer 3.1 was done. On the other hand, the in-vitro part of the study comprised stages of robotic microarray chip probe spotting, bacterial DNAs extraction and DNA labeling, hybridization and microarray chip scanning. In wet lab section, different tools and apparatus such as Nexterion® Slide E, Qarraymini spotter, NimbleGen kit, TrayMixTM S4, and Innoscan 710 were used. A DNA microarray chip including 10 long oligo microarray probes was designed and constructed for detection and identification of 10 pathogenic bacteria. The DNA microarray chip was capable to identify all 10 bacterial agents tested simultaneously. The presence of a professional bioinformatician as a probe designer is needed to design appropriate multifunctional microarray probes to increase the accuracy of the outcomes.


Antepartum continuous epidural ropivacaine therapy reduces uterine artery vascular resistance in pre-eclampsia: a randomized, dose-ranging, placebo-controlled study.
No therapy is currently available to improve the reduced uteroplacental blood flow (UPBF) that characterizes pre-eclampsia. We hypothesized that sympathectomy induced by epidural local anaesthesia reduces uterine vascular resistance (which is inversely correlated with UPBF) in pre-eclampsia. Ten pregnant women between 24 and 32 weeks of gestation with pre-eclampsia and uterine artery flow abnormalities were randomized to antepartum continuous epidural therapy (ACET) or control. ACET was initiated by a 5 day dose-ranging trial (ACET-1) of 0.04, 0.06, 0.08, and 0.1% ropivacaine and saline placebo, each at 10 ml h(-1) for 24 h. Doses were randomized and double-blind. Doppler ultrasound indices of vascular resistance were assessed at baseline and after each 24 h dosing period in both uterine arteries. Subsequently, these ACET patients were administered 0.1% ropivacaine until delivery (ACET-2), with one additional randomized double-blind placebo day. Five patients were randomized to ACET. In each patient, one uterine artery exhibited a dose-dependent reduction in vascular resistance (P=0.035), a response that returned to baseline following placebo (P<0.001). The contralateral uterine artery exhibited either increased vascular resistance or no change. In all cases, the uterine artery that responded to ACET had higher baseline resistance than its pair (P=0.043). Baseline right-left difference in resistance between paired uterine arteries was greatly diminished following ACET. Although ACET patients had a mean (sd) duration to delivery of 19 (9) days compared with control 2 (1) days (P=0.008), this should be interpreted with caution because of demographic differences between groups. ACET reduces uterine artery resistance in pre-eclampsia <32 weeks. Uteroplacental re-distribution is a novel observation and warrants further investigation.


Schizophrenia and employment - a review.
Little is known about the extent to which work contributes to the recovery of people with schizophrenia. There is increasing interest in the subject because of new service models and the economic cost of unemployment in people with severe mental illness. A literature search was carried out with the aim of investigating: a). employment rates in schizophrenia and first-episode psychosis and the extent to which they have changed over time; b). the barriers to work; c). the factors associated with being employed among people with schizophrenia; and d). whether employment influences other outcomes in schizophrenia. There are wide variations in reported employment rates in schizophrenia. Most recent European studies report rates between 10 % and 20%, while the rate in the US is less clear. There is a higher level of employment among first-episode patients. The employment rate in schizophrenia appears to have declined over the last 50 years in the UK. Barriers to getting employment include stigma,discrimination, fear of loss of benefits and a lack of appropriate professional help. The most consistent predictor of employment is previous work history. Working is correlated with positive outcomes in social functioning, symptom levels, quality of life and self esteem, but a clear causal relationship has not been established. Very low employment rates are not intrinsic to schizophrenia, but appear to reflect an interplay between the social and economic pressures that patients face, the labour market and psychological and social barriers to working.


Actual and ideal services in acute care and rehabilitation for relatives post-stroke from three perspectives: Relatives, stroke clients and health professionals.
To document the gap between actual and desired ideal services for relatives post-stroke from three perspectives: relatives, stroke clients and health professionals. A two-phase qualitative design and consisting of in-depth interviews (Phase 1) and 3 focus groups (Phase 2). The interview sample consisted of 25 relatives (mean age 53.4 (standard deviation 12.7); women = 21/25) and 16 individuals with a first stroke (mean age = 55.7 (standard deviation 11.2); women = 7/16). The focus group sample size varied from 5 to 7 participants. An interview guide validated by experts was used. Audio content was transcribed verbatim and rigorously analyzed by two team members. Services received by relatives are diversified, and relatives' perceptions range from receiving no services to being satisfied with services received. Even when participants were satisfied, ideal services were still desired: they would have liked to receive services earlier and without having to seek. Four main factors emerged as influencing the amount and quality of services received, including the individual's ability to seek. A gap remains between actual and ideal services for relatives post-stroke. It is crucial to legitimized relatives' role as clients and to systematically assess the patient's social environment in order to provide services in accordance with needs.


Surgical steps of total laparoscopic hysterectomy: Part 1: Benign disease by the European Society for Gynaecological Endoscopy (ESGE)1.
Laparoscopic hysterectomy, which is associated with well-documented benefits compared to an abdominal approach, should be the surgical treatment choice for certain gynaecological conditions. The aim of this document is to provide recommendations on the technical aspects of this procedure. The European Society for Gynaecological Endoscopy (ESGE) has established a working group with the goal of standardizing the surgical steps of total laparoscopic hysterectomy for benign disease, which is one of the most commonly performed major surgical procedures in Gynaecology. A properly equipped surgical theatre and the correct positioning of both the laparoscopic tower and the surgeons are of utmost importance and insure that the surgical procedure can be performed uneventfully.The incorporation of standardized surgical steps in routine practice should help to ensure an optimal surgical outcome based on the available scientific evidence as reviewed by expert surgeons. The description of the surgical steps of total laparoscopic hysterectomy and its adoption by minimally invasive gynaecologists will make this approach safer.


Systemic stiffening of mouse tail tendon is related to dietary advanced glycation end products but not high-fat diet or cholesterol.
Tendon pathology is related to metabolic disease and mechanical overloading, but the effect of metabolic disease on tendon mechanics is unknown. This study investigated the effect of diet and apolipoprotein E deficiency (ApoE(-/-)) on mechanical properties and advanced glycation end product (AGE) cross-linking of non-weight-bearing mouse tail tendons. Twenty ApoE(-/-) male mice were used as a model for hypercholesterolemia along with 26 wild-type (WT) mice. One-half of the mice from each group was fed a normal diet (ND) and the other half was fed a high-fat diet (HFD) to induce obesity. All were killed at 40 wk, and tail tendon fascicles were mechanically tested to failure and analyzed for AGEs. Diets were also analyzed for AGEs. ApoE(-/-) mice displayed a 14% increase in plateau modulus compared with WT mice (P < 0.05), whereas HFD mice displayed a 13% decrease in plateau modulus (P < 0.05) and a 12% decrease in total modulus (P < 0.05) compared with ND mice. Tail tendons of HFD mice had significantly lower concentrations of AGEs [carboxymethyllysine (CML): 26%, P < 0.0001; methylglyoxal-derived hydroimidazolone 1 (MG-H1): 15%, P < 0.005; pentosidine: 13%, P < 0.0005]. The HFD had ∼44-fold lower content of CML (P < 0.01), ∼29-fold lower content of carboxyethyllysine (P < 0.005), and ∼16-fold lower content of MG-H1 (P < 0.05) compared with ND. ApoE(-/-) increased, whereas HFD decreased mouse tail tendon stiffness. Dietary AGE content may be a crucial determinant for accumulation of AGE cross-links in tendons and for tissue compliance. The results demonstrate how systemic metabolic factors may influence tendon health.


Reconstructing apology: David Cameron's Bloody Sunday apology in the press.
While there is an acknowledgement in apology research that political apologies are highly mediated, the process of mediation itself has lacked scrutiny. This article suggests that the idea of reconstruction helps to understand how apologies are mediated and evaluated. David Cameron's apology for Bloody Sunday is examined to see how he constructs four aspects of apology: social actors, consequences, categorization, and reasons. The reconstruction of those aspects by British, Unionist, and Nationalist press along with reconstructions made by soldiers in an online forum are considered. Data analysis was informed by thematic analysis and discourse analysis which helped to explore key aspects of reconstruction and how elements of Cameron's apology are altered in subsequent mediated forms of the apology. These mediated reconstructions of the apology allowed their authors to evaluate the apology in different ways. Thus, in this article, it is suggested that the evaluation of the apology by different groups is preceded by a reconstruction of it in accordance with rhetorical goals. This illuminates the process of mediation and helps to understand divergent responses to political apologies.


NUC-1031, use of ProTide technology to circumvent gemcitabine resistance: current status in clinical trials.
Resistance to gemcitabine chemotherapy is common in patients with pancreatic ductal adenocarcinoma (PDAC), biliary tract cancer (BTC) and ovarian cancers (OC), conferring poor survival. Use of ProTide technology led to the development of a 'partially-activated' monophosphorylated gemcitabine compound, termed NUC-1031. NUC-1031 enters cancer cells independent of the human equilibrative nucleoside transporter, does not require deoxycytidine kinase-mediated activation and resists cytidine deaminase-mediated breakdown into toxic by-products. The phase I PRO-001 trial recruited 68 patients with advanced solid tumours; of the 49 patients that had response-evaluable disease, 5 (10%) had a partial response (PR) and 33 (67%) had stable disease (SD). Subsequently, the PRO-002 study assessed the safety and efficacy of NUC-1031 combined with carboplatin for patients with OC (n = 25); preliminary data from this study reported one (4%) unconfirmed complete response (CR), 8 (35%) PRs and 13 (57%) patients with SD, the final outcome data are awaited. The ABC-08 trial for advanced BTC assessed safety and efficacy of NUC-1031 combined with cisplatin; 14 patients were recruited with a 50% objective response rate in the intention to treat population at interim analysis. ACELARATE, the phase III trial in first-line advanced PDAC comparing NUC-1031 to gemcitabine monotherapy, recruited 200 patients but has been paused for futility analysis. Early studies demonstrate NUC-1031 is well tolerated with favourable pharmacokinetic profiles. NUC-1031 use in PDAC remains unclear, but encouraging results of disease control in BTC and OC has prompted phase II and III trial development. NuTide 121, is a phase III trial comparing cisplatin-NUC 1031 combination to the standard of care cisplatin-gemcitabine and recruitment is ongoing. Recruiting trials and mature data from existing studies will help inform on the impact of NUC-1031 on patient survival over standard gemcitabine.


Desert dust: an unrecognized source of dangerous air pollution?
Particulate matter (PM10) air pollution is associated with respiratory and cardiovascular morbidity, and mortality. A recent systematic review pointed toward the fine particle fraction (PM2.5) rather than the coarse fraction (PM2.5-10) 2.5 and PM2.5-10as being responsible for increased death rates. With this background, the report by Perez et al that windblown Saharan desert dust causes increased mortality in Barcelona, raises concern over possible underestimation of toxicity from coarse particles coming from desert sources. This may be of concern for large areas of the globe that periodically encounter high levels of windblown desert dust and warrants further attention.


Somatosensory cortical activity in relation to arm posture: nonuniform spatial tuning.
1. Single unitary activity in primate somatosensory cortex (SI) was recorded while monkeys maintained a range of static arm postures. Unit discharge was related to parameters defining the posture of the arm by multiple linear regression techniques. 2. Two monkeys were trained to grasp a manipulandum presented at locations distributed throughout their workspace. The discharge of single units in SI was recorded for 3 s while the monkeys maintained contact with the manipulandum and the mean discharge rate over this hold time was related to the location of the hand and to the shoulder and elbow joint angles of the arm. 3. Unitary activity of 171 neurons in the proximal arm region of areas 3, 1, and 2 was recorded during the task. Of the total, 78 neurons had activity that varied with the location of the hand in space. Neuronal discharge typically varied monotonically with the target location, reaching a maximum at the borders of the work-space. The discharge rate in most of these neurons varied with both shoulder and elbow angles. 4. Discharge rate was related to the hand's location along three axes by means of a polynomial fit. In approximately half of the neurons, activity varied significantly only for displacements along a single axis in space. However, many neurons exhibited nonlinear relations between hand location along this preferred axis and discharge rate. Discharge rate did not vary for displacements of the hand in the plane perpendicular to this preferred axis (null plane). 5. In other neurons, discharge rate varied for hand displacements in a plane, i.e., along two perpendicular axes. Displacements of the hand along the axis perpendicular to this plane (null axis) did not affect the discharge rate. In only a small minority of neurons did discharge rate vary for hand displacements along all three axes in space. 6. The distribution of the sensitivity of the neural population to hand displacements along arbitrary directions in space was not uniform. On average, hand displacement along a vertical axis led to the smallest modulation of neural discharge, and displacement of the hand along the anteroposterior direction led to the largest modulation of activity.


Path integration of head direction: updating a packet of neural activity at the correct speed using neuronal time constants.
A key question in understanding the neural basis of path integration is how individual, spatially responsive, neurons may self-organize into networks that can, through learning, integrate velocity signals to update a continuous representation of location within an environment. It is of vital importance that this internal representation of position is updated at the correct speed, and in real time, to accurately reflect the motion of the animal. In this article, we present a biologically plausible model of velocity path integration of head direction that can solve this problem using neuronal time constants to effect natural time delays, over which associations can be learned through associative Hebbian learning rules. The model comprises a linked continuous attractor network and competitive network. In simulation, we show that the same model is able to learn two different speeds of rotation when implemented with two different values for the time constant, and without the need to alter any other model parameters. The proposed model could be extended to path integration of place in the environment, and path integration of spatial view.


In vitro human metabolism of the flame retardant resorcinol bis(diphenylphosphate) (RDP).
Resorcinol bis(diphenylphosphate) (RDP) is widely used as a flame retardant in electrical/electronic products and constitutes a suitable alternative to decabrominated diphenyl ether. Due to its toxicity and its recently reported ubiquity in electronics and house dust, there are increasing concerns about human exposure to this emerging contaminant. With the aim of identifying human-specific biomarkers, the in vitro metabolism of RDP and its oligomers was investigated using human liver microsomes and human liver cytosol. Mono- and dihydroxy-metabolites, together with glucuronidated and sulfated metabolites, were detected. Regarding RDP oligomers, only a hydroxy-metabolite of the dimer could be detected. RDP and its oligomers were also readily hydrolyzed, giving rise to a variety of compounds, such as diphenyl phosphate, para-hydroxy-triphenyl phosphate, and para-hydroxy RDP, which were further metabolized. These degradation products or impurities are possibly of environmental importance in future studies.


Glanzmann thrombasthenia with acute myeloid leukemia successfully treated by bone marrow transplantation.
We report successful treatment by bone marrow transplantation (BMT) in an acute myeloid leukemia (AML) patient with Glanzmann thrombasthenia (GT). Genetic analysis revealed that a novel point mutation in exon 3 of the GPIIb gene led to abnormal splicing resulting in an amino acid substitution and an in-frame deletion of 3 amino acid residues. Expression studies suggested a rapid degradation of the uncomplexed protein within the cells. Induction therapy for AML was performed with frequent platelet transfusions because of the patient's severe hemorrhagic manifestations. In the second remission, the patient was successfully treated by BMT from an HLA-matched unrelated donor. Platelet function returned to normal, and the GT phenotype completely disappeared. Our experience suggests that BMT is a curative therapeutic strategy for GT. Furthermore, we believe this study is the first to demonstrate that engraftment after BMT for AML can be determined by monitoring the congenital genetic defect of GT.


Risk of coronary artery disease in adults with congenital heart disease: A comparison with the general population.
Coronary artery disease (CAD) will increasingly determine outcome in the aging adult congenital heart disease (CHD) population. We aimed to determine sex-specific incidence of CAD in adult CHD patients throughout adulthood, compared to the general population. We followed 11,723 adult CHD patients (median age 33 years; 49% male; 57% mild, 34% moderate, 9% severe CHD) from the Dutch CONCOR registry, and two age-sex-matched persons per patient from the general population for first CAD event in national registers (period 2002-2012). Incidence rates were estimated using smoothed hazard functions. CAD risk during follow-up, stratified by CHD severity, was compared using proportional subdistribution hazards regression. In ACHD patients, 103 CAD events (43 women) occurred over 60,456 person-years. Rates per 1000person-years increased from 0.3(95% confidence interval: 0.1-0.6) at age 20 to 5.8(3.7-8.9) at 70 years in female, and from 0.5(0.3-1.0) to 7.8(5.1-11.8) in male patients. Compared to the general population, relative risk was 12.0(2.5-56.3) in women and 4.6(1.7-12.1) in men aged 20 years. Relative risk declined with age, remaining significant up to age ~65 years in women and ~50 years in men. In patients with mild, moderate and severe CHD, CAD risk was 1.3(0.9-1.9), 1.6(1.0-2.5) and 2.9(1.3-6.9) times increased compared to the general population, respectively. We found increased CAD risk in adult CHD patients, with greater relative risk at younger age, in women and those with more severe CHD. These results underline the importance of screening for and treatment of CAD risk factors in these patients.


Effect of the Polydispersity of a Colloidal Drop on Drying Induced Stress as Measured by the Buckling of a Floating Sheet.
We study the stress developed during the drying of a colloidal drop of silica nanoparticles. In particular, we use the wrinkling instability of a thin floating sheet to measure the net stress applied by the deposit on the substrate and we focus on the effect of the particle polydispersity. In the case of a bidisperse suspension, we show that a small number of large particles substantially decreases the expected stress, which we interpret as the formation of lower hydrodynamic resistance paths in the porous material. As colloidal suspensions are usually polydisperse, we show for different average particle sizes that the stress is effectively dominated by the larger particles of the distribution and not by the average particle size.


Limitation of Trypanosoma brucei parasitaemia results from density-dependent parasite differentiation and parasite killing by the host immune response.
In the bloodstream of its mammalian host, the "slender" form of Trypanosoma brucei replicates extracellularly, producing a parasitaemia. At high density, the level of parasitaemia is limited at a sublethal level by differentiation to the non-replicative "stumpy" form and by the host immune response. Here, we derive continuous time equations to model the time-course, cell types and level of trypanosome parasitaemia, and compare the best fits with experimental data. The best fits that were obtained favour a model in which both density-dependent trypanosome differentiation and host immune response have a role in limiting the increase of parasites, much poorer fits being obtained when differentiation and immune response are considered independently of one another. Best fits also favour a model in which the slender-to-stumpy differentiation progresses in a manner that is essentially independent of the cell cycle. Finally, these models also make the prediction that the density-dependent trypanosome differentiation mechanism can give rise to oscillations in parasitaemia level. These oscillations are independent of the immune system and are not due to antigenic variation.


Hypoxia-inducible factor-1 alpha in non small cell lung cancer: relation to growth factor, protease and apoptosis pathways.
Hypoxia-inducible factor (HIF)-1 alpha is the regulatory subunit of HIF-1 that is stabilized under hypoxic conditions. Under different circumstances, HIF-1 alpha may promote both tumorigenesis and apoptosis. There is conflicting data on the importance of HIF-1 alpha as a prognostic factor. This study evaluated HIF-1 alpha expression in 172 consecutive patients with stage I-IIIA non small cell lung cancer (NSCLC) using standard immunohistochemical techniques. The extent of HIF-1 alpha nuclear immunostaining was determined using light microscopy and the results were analyzed using the median (5%) as a low cut-point and 60% as a high positive cut-point. Using the low cut-point, positive associations were found with epidermal growth factor receptor (EGFR; p = 0.01), matrix metalloproteinase (MMP)-9 (p = 0.003), membranous (p < 0.001) and perinuclear (p = 0.004) carbonic anhydrase (CA) IX, p53 (p = 0.008), T-stage (p = 0.042), tumor necrosis (TN; p < 0.001) and squamous histology (p < 0.001). No significant association was found with Bcl-2 or either N- or overall TMN stage or prognosis. When the high positive cut-point was used, HIF-1 alpha was associated with a poor prognosis (p = 0.034). In conclusion, the associations with EGFR, MMP-9, p53 and CA IX suggest that these factors may either regulate or be regulated by HIF-1 alpha. The association with TN and squamous-type histology, which is relatively more necrotic than other NSCLC types, reflects the role of hypoxia in the regulation of HIF-1 alpha. The prognostic data may reflect a change in the behavior of HIF-1 alpha in increasingly hypoxic environments.


Kinematic and Gait Similarities between Crawling Human Infants and Other Quadruped Mammals.
Crawling on hands and knees is an early pattern of human infant locomotion, which offers an interesting way of studying quadrupedalism in one of its simplest form. We investigate how crawling human infants compare to other quadruped mammals, especially primates. We present quantitative data on both the gait and kinematics of seven 10-month-old crawling infants. Body movements were measured with an optoelectronic system giving precise data on 3-dimensional limb movements. Crawling on hands and knees is very similar to the locomotion of non-human primates in terms of the quite protracted arm at touch-down, the coordination between the spine movements in the lateral plane and the limbs, the relatively extended limbs during locomotion and the strong correlation between stance duration and speed of locomotion. However, there are important differences compared to primates, such as the choice of a lateral-sequence walking gait, which is similar to most non-primate mammals and the relatively stiff elbows during stance as opposed to the quite compliant gaits of primates. These finding raise the question of the role of both the mechanical structure of the body and neural control on the determination of these characteristics.


The Canadian childhood nephrotic syndrome (CHILDNEPH) study: report on mid-study feasibility, recruitment and main measures.
To assess reasons for continuing practice variation in the management of childhood nephrotic syndrome despite expert reviews and guidelines, we are conducting a longitudinal cohort study in children with glucocorticoid sensitive nephrotic syndrome. Objectives of this mid-study report are to describe patient and physician recruitment characteristics, glucocorticoid prescriptions, use of second line agents, biopsy practices, and adherence to study protocol. Children with new onset nephrotic syndrome and providers are being recruited from all 12 pediatric nephrology centres across Canada with > 2½ years follow-up. Data collection points of observation are over a minimum 36 months. Details of prescribed glucocorticoids and of all second line agents used during treatment are being collected. All relapses are being recorded with time to urinary remission of proteinuria. To date, 243 patients (57.1% male) from 12 centres were included. Median number of patients per centre was 29 (range 2-45), and median age of cohort was 7.3 (IQR 4.2) at enrollment. Forty-eight physicians were recruited, median 5 (range 2-8) per site. Median number of relapses per patient year of follow-up was 2.1 (IQR 4). Cumulative dose variability of glucocorticoids prescribed per episode of proteinuria and length of treatment was observed between participating centres. The Canadian pediatric nephrology community established a longitudinal childhood nephrotic syndrome cohort study that confirms ongoing practice variability. The study will help to evaluate its impact on patient outcomes, and facilitate clinical trial implementation in nephrotic syndrome.


Hypothermia and rewarming after hypothermic exposure alter venous relaxation.
Hypothermia has pathophysiological consequences on endothelial and smooth muscle cell function. This study investigates the impact on venous relaxation of hypothermia and rewarming following hypothermic exposure. In vitro isometric relaxation responses of norepinephrine precontracted rabbit external jugular veins to a panel of endothelium-dependent and -independent agonists were assessed in controls at 37 degrees C, and in an experimental group after cooling to 20 degrees C and after rewarming to 37 degrees C. On cooling, the endothelium-dependent responses to acetylcholine became multiphasic with initial contraction at low concentrations followed by relaxation at higher concentrations, the maximum of which was significantly diminished compared to controls. Incubation with indomethacin did not affect this response. Rewarming re-established a monophasic dose-dependent acetylcholine induced relaxation response but the maximal response was significantly augmented. This augmentation in relaxation on rewarming could be prevented by preincubation with indomethacin. The maximal response to calcium ionophore was reduced at 20 degrees C and augmented upon rewarming to 37 degrees C. All veins relaxed in a dose-dependent manner to the non-endothelium-dependent agonists forskolin and sodium nitroprusside; the maximal responses were significantly reduced at 20 degrees C and returned to normal upon rewarming. This study suggests that short-term exposure of venous tissue to hypothermia impairs the vessel's ability to produce endothelium-dependent relaxation. Rewarming does not re-establish normal endothelium-dependent relaxation but results in an enhanced, partially indomethacin-sensitive, response which appears to be independent of changes in non-endothelium-dependent mediated relaxation.


Nucleation and Conformality of Iridium and Iridium Oxide Thin Films Grown by Atomic Layer Deposition.
Nucleation and conformality are important issues, when depositing thin films for demanding applications. In this study, iridium and iridium dioxide (IrO2) films were deposited by atomic layer deposition (ALD), using five different processes. Different reactants, namely, O2, air, consecutive O2 and H2 (O2 + H2), and consecutive O3 and H2 (O3 + H2) pulses were used with iridium acetylacetonate [Ir(acac)3] to deposit Ir, while IrO2 was deposited using Ir(acac)3 and O3. Nucleation was studied using a combination of methods for film thickness and morphology evaluation. In conformality studies, microscopic lateral high-aspect-ratio (LHAR) test structures, specifically designed for accurate and versatile conformality testing of ALD films, were used. The order of nucleation, from the fastest to the slowest, was O2 + H2 > air ≈ O2 > O3 > O3 + H2, whereas the order of conformality, from the best to the worst, was O3 + H2 > O2 + H2 > O2 > O3. In the O3 process, a change in film composition from IrO2 to metallic Ir was seen inside the LHAR structures. Compared to the previous reports on ALD of platinum-group metals, most of the studied processes showed good to excellent results.


Reference ranges for lymphocyte subsets in healthy adult male Omanis.
To determine the reference ranges of lymphocyte subsets in serologically HIV-seronegative healthy male adults in Oman. A cohort, of 118 healthy male blood donors ranging in age from 18-51 years, was included in the study. The average age was 25 years. Blood samples collected into tubes containing ethylene-diamine-tetra acetic acid were investigated for lymphocyte subsets using flow cytometer. This study was conducted in the Immunology Laboratory of the Sultan Qaboos University, College of Medicine and Health Sciences, Muscat, Oman during the year 2006. For the 118 males investigated, the mean percentage and absolute values of the lymphocyte subsets were as follows: CD3: 68.53 +/- 7.5%, 1701 +/- 489 cells/microliter; CD4: 40.4 +/- 6.5%, 1006 +/- 319 cells/microliter; CD8: 25.8 +/- 5.9%, 638 +/- 225 cells/microliter; CD19: 13.7 +/- 4.7%, 349 +/- 158 cells/microliter, and CD56: 12.2 +/- 6.7%, 308 +/- 204 cells/microliter. The ratio of CD4/CD8 was 1.6. Immunophenotyping has been used to establish reference values of lymphocyte subsets in normal healthy adult males in Oman. The Omani male reference values obtained in this study show wide variations compared with kits values previously used as a reference.


Size-dependent nanoscale kirkendall effect during the oxidation of nickel nanoparticles.
The transformation of Ni nanoparticles (NPs) of different sizes (average diameters of 9, 26, and 96 nm) during oxidation to hollow (single void) or porous (multiple voids) NiO through the nanoscale Kirkendall effect was observed by transmission electron microscopy. Samples treated for 1-4 h at 200-500 degrees C show that the structures of the completely oxidized NPs do not depend on the temperature, but oxidation proceeds more quickly at elevated temperatures. For the Ni/NiO system, after formation of an initial NiO shell (of thickness approximately 3 nm), single or multiple voids nucleate on the inner surface of the NiO shell, and the voids grow until conversion to NiO is complete. Differences in the void formation and growth processes cause size-dependent nanostructural evolution: For 9 and 26 nm NPs, a single void forms beneath the NiO shell, and the void grows by moving across the NP while conversion to NiO occurs opposite the site where the void initially formed. Because of the differences in the Ni/NiO volume ratios for the 9 and 26 nm NPs when the void first forms, they have distinct nanostructures: The 9 nm NPs form NiO shells that are nearly radially symmetric, while there is a pronounced asymmetry in the NiO shells for 26 nm NPs. By choosing an intermediate oxidation temperature and varying the reaction time, partially oxidized Ni(core)/NiO(shell) NPs can be synthesized with good control. For 96 nm NPs, multiple voids form and grow, which results in porous NiO NPs.


Tropism, replication competence, and innate immune responses of influenza virus: an analysis of human airway organoids and ex-vivo bronchus cultures.
Human airway organoids are three-dimensional cultures derived from stem cells, which self-organise in ex-vivo conditions to form so-called mini-airways. The cellular morphology of these cultures is physiologically similar to the human airway, with cilia, goblet cells, and club cells facing the inner lumen and basal cells situated at the outer layer. The aim of this study was to compare replication competence, tissue tropism, and host responses elicited by human and avian strains of influenza A virus in ex-vivo human bronchus and human airway organoids. Between Sept 29, 2016, and Jan 4, 2017, we obtained ex-vivo cultures of the human bronchus and cultured human airway organoids from lung stem cells obtained from human lung tissues removed as part of the routine clinical care of patients undergoing surgical resection at the Department of Cardiothoracic Surgery, University of Hong Kong, Queen Mary Hospital, Hong Kong. We compared viral replication competence, tissue tropism, and cytokine and chemokine induction of avian influenza A viruses isolated from humans (Sh2/H7N9, H5N1/483, H5N6/39715), and human H1N1pdm/415742 in airway organoids and ex-vivo bronchus explant cultures. Virus tropism and replication kinetics of human and avian influenza A viruses in human airway organoids mimicked those found in ex-vivo cultures of human bronchus explants. In both airway organoids and bronchus explants, influenza A H1N1 subtype (H1N1) and avian influenza A H7N9 viruses replicated to significantly higher titres than did the highly pathogenic avian influenza (HPAI) H5N1, whereas HPAI H5N6 replication was moderate. H1N1, H7N9, and H5N6 viruses infected ciliated cells and goblet cells, but not basal cells in both airway organoids and bronchus explants. The expression of cytokines, interleukin 6, and interferon β, and the chemokine regulated-on-activation, normal T-cell expressed and secreted, was significantly higher in human airway organoids infected with HPAI H5N1 virus than H1N1pdm/415742, Sh2/H7N9, and H5N6/39715 viruses, and the expression of monocyte chemoattractant protein-1 was significantly higher in human organoids infected with HPAI H5N1 virus than H1N1pdm/415742 and Sh2/H7N9 viruses. Human airway organoid cultures provided results that were comparable to those observed in human ex-vivo bronchus cultures, and thus provide an alternative physiologically relevant experimental model for investigating virus tropism and replication competence that could be used to assess the pandemic threat of animal influenza viruses. US National Institute of Allergy and Infectious Diseases, Research Grants Council of the Hong Kong Special Administrative Region.


Acute renal failure - definition, outcome measures, animal models, fluid therapy and information technology needs: the Second International Consensus Conference of the Acute Dialysis Quality Initiative (ADQI) Group.
There is no consensus definition of acute renal failure (ARF) in critically ill patients. More than 30 different definitions have been used in the literature, creating much confusion and making comparisons difficult. Similarly, strong debate exists on the validity and clinical relevance of animal models of ARF; on choices of fluid management and of end-points for trials of new interventions in this field; and on how information technology can be used to assist this process. Accordingly, we sought to review the available evidence, make recommendations and delineate key questions for future studies. We undertook a systematic review of the literature using Medline and PubMed searches. We determined a list of key questions and convened a 2-day consensus conference to develop summary statements via a series of alternating breakout and plenary sessions. In these sessions, we identified supporting evidence and generated recommendations and/or directions for future research. We found sufficient consensus on 47 questions to allow the development of recommendations. Importantly, we were able to develop a consensus definition for ARF. In some cases it was also possible to issue useful consensus recommendations for future investigations. We present a summary of the findings. (Full versions of the six workgroups' findings are available on the internet at http://www.ADQI.net) Despite limited data, broad areas of consensus exist for the physiological and clinical principles needed to guide the development of consensus recommendations for defining ARF, selection of animal models, methods of monitoring fluid therapy, choice of physiological and clinical end-points for trials, and the possible role of information technology.


A hybrid biomimetic nanomatrix composed of electrospun polycaprolactone and bioactive peptide amphiphiles for cardiovascular implants.
Current cardiovascular therapies are limited by the loss of endothelium, restenosis and thrombosis. The goal of this study was to develop a biomimetic hybrid nanomatrix that combined the unique properties of electrospun polycaprolactone (ePCL) nanofibers with self-assembled peptide amphiphiles (PAs). ePCL nanofibers have interconnected nanoporous structures, but are hampered by a lack of surface bioactivity to control cellular behavior. It has been hypothesized that PAs could self-assemble onto the surface of ePCL nanofibers and endow them with the characteristic properties of native endothelium. The PAs, which comprised hydrophobic alkyl tails attached to functional hydrophilic peptide sequences, contained enzyme-mediated degradable sites coupled to either endothelial cell-adhesive ligands (YIGSR) or polylysine (KKKKK) nitric oxide (NO) donors. Two different PAs (PA-YIGSR and PA-KKKKK) were successfully synthesized and mixed in a 90:10 (YK) ratio to obtain PA-YK. PA-YK was reacted with pure NO to develop PA-YK-NO, which was then self-assembled onto ePCL nanofibers to generate a hybrid nanomatrix, ePCL-PA-YK-NO. Uniform coating of self-assembled PA nanofibers on ePCL was confirmed by transmission electron microscopy. Successful NO release from ePCL-PA-YK-NO was observed. ePCL-YK and ePCL-PA-YK-NO showed significantly increased adhesion of human umbilical vein endothelial cells (HUVECs). ePCL-PA-YK-NO also showed significantly increased proliferation of HUVECs and reduced smooth muscle cell proliferation. ePCL-PA-YK-NO also displayed significantly reduced platelet adhesion compared with ePCL, ePCL-PA-YK and a collagen control. These results indicate that this hybrid nanomatrix has great potential application in cardiovascular implants.


The influence of weather on community gastroenteritis in Australia.
Infectious gastroenteritis is a common illness in Australia as elsewhere. Data from a year-long national gastroenteritis survey in 2001-2002 showed that gastroenteritis was more common in the northern and hotter part of Australia. These data were used to quantify associations between local weather variables and gastroenteritis in people aged >5 years while controlling for socioeconomic status. A distributed lag model was used to examine the influence of weather over a period of days prior to an event and the maximal effect was found at a lag of 2-5 days. The total effect over the preceding week indicated a relative increase from baseline in the probability of gastroenteritis of 2·48% (95% CI 1·01-3·97) for each degree rise (°C) over that period. Given the very high burden of gastroenteritis, this represents a substantial effect at the population level and has relevance for health predictions due to climate change.


Highly unusual interpenetration isomers of electroactive nickel bis(dithiolene) coordination frameworks.
The metalloligand [Ni(pedt)2](-) (pedt = 1-(pyridine-4-yl)ethylene-1,2-dithiolate) has been incorporated into two multi-dimensional structures for the first time. These coordination frameworks represent highly unusual interpenetration isomers and exhibit solid state redox and optical properties that reflect the electronically delocalised nature of the metalloligand.


Principles and engineering of antibody folding and assembly.
Antibodies are uniquely suited to serve essential roles in the human immune defense as they combine several specific functions in one hetero-oligomeric protein. Their constant regions activate effector functions and their variable domains provide a stable framework that allows incorporation of highly diverse loop sequences. The combination of non-germline DNA recombination and mutation together with heavy and light chain assembly allows developing variable regions that specifically recognize essentially any antigen they may encounter. However, this diversity also requires tailor-made mechanisms to guarantee that folding and association of antibodies is carefully this diversity also requires tailor-made mechanisms to guarantee that folding and association of antibodies is carefully controlled before the protein is secreted from a plasma cell. Accordingly, the generic immunoglobulin fold β-barrel structure of antibody domains has been fine-tuned during evolution to fit the different requirements. Work over the past decades has identified important aspects of the folding and assembly of antibody domains and chains revealing domain specific variations of a general scheme. The most striking is the folding of an intrinsically disordered antibody domain in the context of its partner domain as the basis for antibody assembly and its control on the molecular level in the cell. These insights have not only allowed a better understanding of the antibody folding process but also provide a wealth of opportunities for rational optimization of antibody molecules. In this review, we summarize current concepts of antibody folding and assembly and discuss how they can be utilized to engineer antibodies with improved performance for different applications. This article is part of a Special Issue entitled: Recent advances in the molecular engineering of antibodies.


A Review of Starch Biosynthesis in Relation to the Building Block-Backbone Model.
Starch is a water-insoluble polymer of glucose synthesized as discrete granules inside the stroma of plastids in plant cells. Starch reserves provide a source of carbohydrate for immediate growth and development, and act as long term carbon stores in endosperms and seed tissues for growth of the next generation, making starch of huge agricultural importance. The starch granule has a highly complex hierarchical structure arising from the combined actions of a large array of enzymes as well as physicochemical self-assembly mechanisms. Understanding the precise nature of granule architecture, and how both biological and abiotic factors determine this structure is of both fundamental and practical importance. This review outlines current knowledge of granule architecture and the starch biosynthesis pathway in relation to the building block-backbone model of starch structure. We highlight the gaps in our knowledge in relation to our understanding of the structure and synthesis of starch, and argue that the building block-backbone model takes accurate account of both structural and biochemical data.


Bacterial seed endophyte community of annual plants modulated by plant photosynthetic pathways.
Climate change is predicted to have adverse impacts on terrestrial ecosystems and uncertainties exist on how these systems will respond. Terrestrial plant ecosystems can be divided by how they fix atmospheric carbon- C3, C4 and CAM photosynthesis pathways. However, as for now, no clear answers could be given regarding the future global repartition of the C3, C4 and CAM plants. As seeds are the reproductive and dispersal unit of the plants and endophytes play a central role in their preservation; here it is suggested that a better knowledge regarding the seeds endophytic community is needed when studying the future repartition of C3, C4 and CAM plant seeds. Bacterial endophyte communities inhabiting seeds belonging to C3, C4 and CAM annual plants were analysed by culture-dependent methods and 16S rRNA gene sequencing. Results indicated there were differences in the relative abundance of bacterial phyla within and across all photosynthetic pathways. Indicating some level of niche partitioning, and each of the three photosynthetic pathways could be characterized by a specific endophytic composition of Firmicutes, corresponding to the adaptation capacity of each group. We successfully identified resistant species of endophytes in the Firmicutes phylum of C4 and CAM plant seeds. Those bacteria are known for being involved in thermal regulation and plant protection through enzymes and antibiotic synthesis and match the strong adaptation capacity of C4 and CAM plants. Overall, this study suggests that there is a plant-mediated selection of the seed microbiome and these symbionts could potentially confer additional benefits to the seed.


Plasmodium falciparum resistant to chloroquine and to pyrimethamine in Comoros.
We report the outcome of chloroquine treatment and the prevalence of mutations at codon 86 of the pfmdr1 gene, at codon 76 of the pfcrt gene, and at codon 108 of the pfdhfr gene in clinical isolates of Plasmodium falciparum collected from 30 children under 10 years of age living in the Comoros Union. This in vivo study was carried out in February and March 2001 in Moroni. Chloroquine treatment failed in 23 children (76.6%; 95% confidence interval: 57.7 to 90.1%). Subsequent genotyping showed that all P. falciparum isolates (100%) harboured a tyrosine residue at position 86 in pfMDR1. 83.3% (25/30) of these isolates harboured a mutation at position 76 in pfCRT and half (15/30) of these isolates also harboured a mutation at position 108 in pfDHFR. Chloroquine resistance is a real concern in the Comoros Union. The prevalence of pfDHFR mutant parasites is alarming. The alternative drugs proposed as a replacement for chloroquine as first-line treatment in Comoros, and the strategy to monitor the drug susceptibility of Plasmodium sp in this part of the Indian Ocean sub-region are discussed.


Spectroscopic and diffraction study of uranium speciation in contaminated vadose zone sediments from the Hanford site, Washington state.
Contamination of vadose zone sediments under tank BX-102 at the Hanford site, Washington, resulted from the accidental release of 7-8 metric tons of uranium dissolved in caustic aqueous sludge in 1951. We have applied synchrotron-based X-ray spectroscopic and diffraction techniques to characterize the speciation of uranium in samples of these contaminated sediments. UIII-edge X-ray absorption fine structure (XAFS) spectroscopic studies demonstrate that uranium occurs predominantly as a uranium(VI) silicate from the uranophane group of minerals. XAFS cannot distinguish between the members of this mineral group due to the near identical local coordination environments of uranium in these phases. However, these phases differ crystallographically, and can be distinguished using X-ray diffraction (XRD) methods. As the concentration of uranium was too low for conventional XRD to detect these phases, X-ray microdiffraction (microXRD) was used to collect diffraction patterns on approximately 20 microm diameter areas of localized high uranium concentration found using microscanning X-ray fluorescence (microSXRF). Only sodium boltwoodite, Na(UO2)(SiO3OH) x 1.5H20, was observed; no other uranophane group minerals were present. Sodium boltwoodite formation has effectively sequestered uranium in these sediments under the current geochemical and hydrologic conditions. Attempts to remediate the uranium contamination will likely face significant difficulties because of the speciation and distribution of uranium in the sediments.


A new malaria vector in Africa: Predicting the expansion range of Anopheles stephensi and identifying the urban populations at risk.
In 2012, an unusual outbreak of urban malaria was reported from Djibouti City in the Horn of Africa and increasingly severe outbreaks have been reported annually ever since. Subsequent investigations discovered the presence of an Asian mosquito species; Anopheles stephensi, a species known to thrive in urban environments. Since that first report, An. stephensi has been identified in Ethiopia and Sudan, and this worrying development has prompted the World Health Organization (WHO) to publish a vector alert calling for active mosquito surveillance in the region. Using an up-to-date database of published locational records for An. stephensi across its full range (Asia, Arabian Peninsula, Horn of Africa) and a set of spatial models that identify the environmental conditions that characterize a species' preferred habitat, we provide evidence-based maps predicting the possible locations across Africa where An. stephensi could establish if allowed to spread unchecked. Unsurprisingly, due to this species' close association with man-made habitats, our maps predict a high probability of presence within many urban cities across Africa where our estimates suggest that over 126 million people reside. Our results strongly support the WHO's call for surveillance and targeted vector control and provide a basis for the prioritization of surveillance.


A Comparative Study of the C-Reactive Protein and the ST-Score (ECG) as Prognostic Indicators in Acute Myocardial Infarction in a Rural Resource-Constrained Hospital Setting in Central India: A Cross-Sectional Study.
The electrocardiogram remains a crucial tool in identification of acute myocardial infarction (AMI). High sensitivity C-reactive protein (hs-CRP) has been found to be strong predictor of coronary artery disease (CAD) and future cardiovascular events. The aim of the present study was to compare hs-CRP and ST-Score (STS) as prognostic indicator in acute ST segment elevated myocardial infarction (STEMI) in central rural India. Cross sectional study, rural hospital in central India. In all patients of STEMI, STS, ST index and hs-CRP were measured on admission and serially. The Primary outcome was all cause mortality at 30 days. Secondary outcome were heart failure, life threatening arrhythmia, cardiogenic shock, re-infarction, hospital stay and re-admission. We used Student's t test to compare means, Chi-square test to compare proportions and Mann Whitney test to compare medians. P value <0.05 will be considered significant. Crude odds ratios were computed to assess the strength of association between risk factors and independent variable along with 95% confidence intervals. STS was significantly higher in patients with poor outcome, when compared with good outcome (20.27mm vs.12.47mm, P = 0.002). On multivariate regression model STS was significant predictor of composite outcome events (OR = 2.74; 95% [CI], 1.46 to 5.17; P = 0.002). The area under the ROC curve was 0.70, with sensitivity of 73.5%, specificity of 58.7%; PPV of 68.3% and NPV of 64.2%. hs-CRP in patients with poor outcome vs. good outcome (6mg/L vs. 3.74mg/L, P = 0.003) and (P = 0.06, 0.85 and 0.12) respectively. STS on admission is independent predictor while hs- CRP is not in resource constrained settings.


Changes in keratin expression during 7,12-dimethylbenz[a]anthracene-induced hamster cheek pouch carcinogenesis.
This study was undertaken to explore the expression of keratins in the hamster cheek pouch carcinogenesis model, using monospecific keratin antibodies and a technique that allows immunoblotting analysis of tissues embedded in paraffin. Changes in keratin expression were correlated with histopathological changes and with the expression of the enzyme gamma-glutamyl transpeptidase. The right cheek pouch of 20 male golden Syrian hamsters was treated with 0.5% 7,12-dimethylbenz[a]anthracene for 16 weeks. As previously described by other laboratories, this treatment resulted in hyperplastic and dysplastic lesions and benign and malignant tumors. The keratins assayed in this study were K14 (Mr 55,000), K1 (Mr 67,000), and K13 (Mr 47,000). The normal hamster cheek pouch epithelium expressed K14 in the basal layer and K13 in the suprabasal and differentiated layers, whereas K1 was not detected by either immunohistochemistry or immunoblotting. Concomitant with 7,12-dimethylbenz[a]anthracene-induced hyperplasia, there were some topographical alterations in the distribution of K14. In this case, K14 was no longer restricted to the basal layer but was also expressed in differentiated cells. The same pattern was also observed in dysplastic lesions and in squamous cell carcinoma. Furthermore, expression of the K13 differentiation-associated keratin was preserved in this hyperplastic epithelium during all the stages of carcinogenesis, including either anaplastic or differentiated areas. In contrast, after 2 weeks of 7,12-dimethylbenz[a]anthracene treatment, K1 expression started as a weak and patchy pattern in suprabasal cells, becoming stronger and more homogeneous at 8 and 16 weeks of treatment. However, K1 was almost absent in squamous cell carcinoma, where only small very well differentiated areas were stained. We also observed gamma-glutamyl transpeptidase-positive foci in earlier stages of carcinogenesis, concomitant with the expression of the K1 keratin. However, it was not possible to find a perfect topographical correspondence between the two events. Alterations in the pattern of keratin expression appear to be a common feature during the development of squamous cell carcinoma in different systems and could be an excellent tool to study carcinogenesis and chemoprevention.


Worldwide relative smoking prevalence among people living with and without HIV.
People living with HIV (PLH) suffer disproportionately from the chronic diseases exacerbated by smoking tobacco. We performed a systematic review and meta-analysis to establish the relative prevalence of smoking among PLH. We included observational studies reporting current smoking rates among PLH and comparators without HIV. We searched Medline, EMBASE, LILACS and SciELO from inception to 31 August 2019. We excluded studies that recruited participants with smoking related illness. We used a random effects model to estimate the odds ratio for current smoking in PLH and people without HIV. We used the Newcastle--Ottawa scale to assess methodological bias. We performed subgroup analysis based on sex and WHO region. We quantified heterogeneity with meta-regression and predictive distributions. PROSPERO registration:CRD42016052608. We identified 6116 studies and included 37. Of 111 258 PLH compared with 10 961 217 HIV-negative participants pooled odds of smoking were 1.64 [(95% confidence interval, 95% CI: 1.45-1.85) (95% prediction interval: 0.66-4.10, I2 = 98.1%)]. Odds for men and women living with HIV were 1.68 [(95% CI: 1.44-1.95) (95% prediction interval: 0.71-3.98, I2 = 91.1%)] and 2.16 [(95% CI: 1.77-2.63) (95% prediction interval: 0.92-5.07, I2 = 81.7%)] respectively. PLH are more likely to be smokers than people without HIV. This finding was true in subgroup analyses of men, women and in four of five WHO regions from which data were available. Meta-regression did not explain heterogeneity, which we attribute to the diversity of PLH populations worldwide. Smoking is a barrier to PLH achieving parity in life expectancy and an important covariate in studies of HIV-associated multimorbidity.


Restoring orbital thinking from real space descriptions: bonding in classical and non-classical transition metal carbonyls.
A combined strategy that unifies our interacting quantum atoms approach (IQA), a chemically intuitive energetic perspective within the quantum theory of atoms in molecules (QTAIM), the domain natural orbitals obtained by the diagonalization of the charge-weighted domain-averaged Fermi hole (DAFH), and the statistical analyses of chemical bonding provided by the electron number distribution functions (EDF) is presented. As shown, it allows for recovering traditional orbital images from the orbital invariant descriptions of QTAIM. It does also provide bonding indices (like bond orders) and bond energetics, all in a per orbital basis, still invariant manner, using a single unified framework. The procedure is applied to show how the Dewar, Chatt, and Ducanson model of bonding in simple transition metal carbonyls may be recovered in the real space. The balance between the number of σ-donated and π-backdonated electrons is negative in classical compounds and positive in non-classical ones. The energetic strength of backdonation is, however, smaller than that of donation. Our technique surpasses conventional orbital models by providing physically sound, quantitative energetics of chemical bonds (or interactions) together with effective one-electron pictures, all for arbitrary wavefunctions.


Double trouble: how big a problem is redundant anaerobic antibiotic coverage in Veterans Affairs medical centres?
To assess the prevalence of, and the variation in, avoidable use of metronidazole in the Veterans Affairs (VA) healthcare system Barcode Medication Administration (BCMA) data were retrospectively assessed for all patients hospitalized between January 2006 and December 2010 in acute-care wards of all VA medical centres (VAMCs) with complete BCMA data and at least 10 acute-care non-intensive care unit (ICU) beds. Potentially avoidable metronidazole days of therapy (DOT) were defined as the administration of metronidazole with another anti-anaerobic antibiotic on the same day for at least two consecutive days during the same hospitalization. Metronidazole was not considered redundant in combination with another anti-anaerobic agent within 28 days after a positive test for Clostridium difficile and during hospitalizations associated with discharge diagnosis codes for cholecystitis or cholangitis. A total of 128 VAMCs satisfied the inclusion criteria. Over the study period there were a total of 782,821 DOT of metronidazole (57.4 DOT per 1000 patient-days), of which 183,267 (23.4%) fulfilled the criteria for avoidable metronidazole DOT. The percentage of avoidable metronidazole DOT remained stable over the study period (22.8% in 2006 and 22.9% in 2010) despite a decrease in overall metronidazole use. There was wide variation in the percentage of avoidable metronidazole DOT among facilities (2010: median 20.3%, IQR 15.3%-29.4%). Piperacillin/tazobactam was the most commonly administered drug on avoidable metronidazole DOT (56.8%). Potentially avoidable use of metronidazole affected about a quarter of all days when metronidazole was given. The combination of metronidazole with piperacillin/tazobactam was particularly common and represents a possible target for antibiotic stewardship interventions.


STARplasty for reconstruction of the burned web space: introduction of an alternative technique for the correction of dorsal neosyndactyly.
Significant hand burns frequently result in dorsal neosyndactyly, despite appropriate wound care, excision/grafting, and occupational therapy. Patients often develop limited abduction, tethered flexion/extension, intrinsic tightness, and inversion of the web space. We present a new technique for neosyndactyly release: the STARplasty (Syndactyly Treatment After Release), named after the appearance of the reconstructed web. We performed a retrospective review of 25 patients who underwent 66 web space reconstructions by a single surgeon, from January 2002 to December 2003. The STARplasty was developed prior to the study period, with the following goals: to use local tissue, negate the need for a skin graft, and permit early range of motion. Combined with longitudinal, transverse, and oblique releases, STARplasty uses a single volar flap, harvested from each sidewall of both proximal phalanges, with 30-degree corners. STARplasty simultaneously deepens and widens the web space while correcting the angle of inclination. Sixteen patients (mean age: 34 years; range: 3-62 years) underwent a total of 33 STARplasties, an average of 37 months after burn injury. Mean area resurfaced per web was 5.2 cm, which contrasts with the 33 non-STARplasty reconstructions, which were used to resurface a mean area of 19.4 cm (P < 0.05) and included 5-flap z-plasties (17), full-thickness skin graft (10), 2-flap z-plasties (3), and advancement flaps (3). No complications occurred in the STARplasty group, including infection, flap loss, dehiscence, nerve injury, or recurrent contracture. All patients had improved function (mean follow-up: 6.7 months; range: 1-18 months). STARplasty is a new, safe, and efficacious technique to correct dorsal neosyndactyly and reconstruct the web space after burn injury.


Efficient Counterselection for Methylococcus capsulatus (Bath) by Using a Mutated pheS Gene.
Methylococcus capsulatus (Bath) is a representative gammaproteobacterial methanotroph that has been studied extensively in diverse research fields. The sacB gene, which encodes levansucrase, causing cell death in the presence of sucrose, is widely used as a counterselectable marker for disruption of a target gene in Gram-negative bacteria. However, sacB is not applicable to all Gram-negative bacteria, and its efficiency for the counterselection of M. capsulatus (Bath) is low. Here, we report the construction of an alternative counterselectable marker, pheS*, by introduction of two point mutations (A306G and T252A) into the pheS gene from M. capsulatus (Bath), which encodes the α-subunit of phenylalanyl-tRNA synthetase. The transformant harboring pheS* on an expression plasmid showed sensitivity to 10 mM p-chloro-phenylalanine, whereas the transformant harboring an empty plasmid showed no sensitivity, indicating the availability of pheS* as a counterselectable marker in M. capsulatus (Bath). To validate the utility of the pheS* marker in counterselection, we attempted to obtain an unmarked mutant of xoxF, a gene encoding the major subunit of Xox methanol dehydrogenase, which we failed to obtain by counterselection using the sacB marker. PCR, immunodetection using an anti-XoxF antiserum, and a cell growth assay in the absence of calcium demonstrated successful disruption of the xoxF gene in M. capsulatus (Bath). The difference in counterselection efficiencies of the markers indicated that pheS* is more suitable than sacB for counterselection in M. capsulatus (Bath). This study provides a new genetic tool enabling efficient counterselection in M. capsulatus (Bath).IMPORTANCE Methanotrophs have long been considered promising strains for biologically reducing methane from the environment and converting it into valuable products, because they can oxidize methane at ambient temperatures and pressures. Although several methodologies and tools for the genetic manipulation of methanotrophs have been developed, their mutagenic efficiency remains lower than that of tractable strains such as Escherichia coli Therefore, further improvements are still desired. The significance of our study is that we increased the efficiency of counterselection in M. capsulatus (Bath) by employing pheS*, which was newly constructed as a counterselectable marker. This will allow for the efficient production of gene-disrupted and gene-integrated mutants of M. capsulatus (Bath). We anticipate that this counterselection system will be utilized widely by the methanotroph research community, leading to improved productivity of methane-based bioproduction and new insights into methanotrophy.


Promoting pollinating insects in intensive agricultural matrices: field-scale experimental manipulation of hay-meadow mowing regimes and its effects on bees.
Bees are a key component of biodiversity as they ensure a crucial ecosystem service: pollination. This ecosystem service is nowadays threatened, because bees suffer from agricultural intensification. Yet, bees rarely benefit from the measures established to promote biodiversity in farmland, such as agri-environment schemes (AES). We experimentally tested if the spatio-temporal modification of mowing regimes within extensively managed hay meadows, a widespread AES, can promote bees. We applied a randomized block design, replicated 12 times across the Swiss lowlands, that consisted of three different mowing treatments: 1) first cut not before 15 June (conventional regime for meadows within Swiss AES); 2) first cut not before 15 June, as treatment 1 but with 15% of area left uncut serving as a refuge; 3) first cut not before 15 July. Bees were collected with pan traps, twice during the vegetation season (before and after mowing). Wild bee abundance and species richness significantly increased in meadows where uncut refuges were left, in comparison to meadows without refuges: there was both an immediate (within year) and cumulative (from one year to the following) positive effect of the uncut refuge treatment. An immediate positive effect of delayed mowing was also evidenced in both wild bees and honey bees. Conventional AES could easily accommodate such a simple management prescription that promotes farmland biodiversity and is likely to enhance pollination services.


The interface between genetics and psychology: lessons from developmental dyslexia.
Developmental dyslexia runs in families, and twin studies have confirmed that there is a substantial genetic contribution to poor reading. The way in which discoveries in molecular genetics are reported can be misleading, encouraging us to think that there are specific genes that might be used to screen for disorder. However, dyslexia is not a classic Mendelian disorder that is caused by a mutation in a single gene. Rather, like many other common disorders, it appears to involve combined effects of many genes and environmental factors, each of which has a small influence, possibly supplemented by rare variants that have larger effects but apply to only a minority of cases. Furthermore, to see clearer relationships between genotype and phenotype, we may need to move beyond the clinical category of dyslexia to look at underlying cognitive deficits that may be implicated in other neurodevelopmental disorders.


Estimation of self-motion duration and distance in rodents.
Spatial orientation and navigation rely on information about landmarks and self-motion cues gained from multi-sensory sources. In this study, we focused on self-motion and examined the capability of rodents to extract and make use of information about own movement, i.e. path integration. Path integration has been investigated in depth in insects and humans. Demonstrations in rodents, however, mostly stem from experiments on heading direction; less is known about distance estimation. We introduce a novel behavioural paradigm that allows for probing temporal and spatial contributions to path integration. The paradigm is a bisection task comprising movement in a virtual reality environment in combination with either timing the duration ran or estimating the distance covered. We performed experiments with Mongolian gerbils and could show that the animals can keep track of time and distance during spatial navigation.