Antibiotic resistance in gram-negative bacilli: a Singapore perspective.
Antibiotic resistance in gram-negative bacilli is an area of increasing importance. This prospective study was performed to survey antibiotic resistance in Escherichia coli (E. coli), Klebsiella spp., Pseudomonas aeruginosa and Acinetobacter spp. over a 1-year period. Non-duplicate isolates of E. coli, Klebsiella spp., P. aeruginosa and Acinetobacter spp. were collected from participating Singapore hospitals during defined collection periods in 2006 and 2007. Confirmatory identification and antibiotic susceptibility testing were performed at Changi General Hospital. Minimum inhibitory concentrations (MIC) to a defined panel of antibiotics were determined using microbroth dilution methods. The presence of extended-spectrum beta lactamases and AmpC beta-lactamases in Enterobacteriaceae was determined by phenotypic methods, and susceptibility results were defined using current breakpoints from the Clinical Laboratory Standards Institute (CLSI). Seven hundred and forty-six gram-negative bacilli were received for testing. Resistance to extended-spectrum cephalosporins was present in a third of Enterobacteriaceae isolates, and extended-spectrum beta-lactamases (ESBL) carriage was present in 19.6% and 30.1% of E. coli and Klebsiella pneumoniae, respectively. AmpC enzymes were also detected in 8.5% and 5.6% of E. coli and K. pneumoniae isolates respectively. All Enterobacteriaceae were susceptible to imipenem and meropenem. The most active antibiotics against P. aeruginosa were amikacin, meropenem and piperacillin-tazobactam. A third of P. aeruginosa showed reduced susceptibility to polymyxin B. Carbapenem resistance was significantly higher in Acinetobacter baumannii (70.5%) than in other Acinetobacter species (25.0%). The most active antibiotic against A. baumannii was polymyxin B. Antibiotic resistance is prevalent in gram-negative bacilli isolated from Singapore hospitals. The MIC testing surveillance programme complemented susceptibility data from wider laboratory-based surveillance, and has revealed emerging mechanisms of antibiotic resistance.


Reactive Astrocytes Contribute to Alzheimer's Disease-Related Neurotoxicity and Synaptotoxicity in a Neuron-Astrocyte Co-culture Assay.
Pathological hallmarks of Alzheimer's disease (AD) include deposition and accumulation of amyloid- β (Aβ), neurofibrillary tangle formation, and neuronal loss. Pathogenesis of presymptomatic disease stages remains elusive, although studies suggest that the early structural and functional alterations likely occur at neuronal dendritic spines. Presymptomatic alterations may also affect different CNS cell types. However, specific contributions of these cell types as cause or consequence of pathology are difficult to study in vivo. There is a shortage of relatively simple, well-defined, and validated in vitro models that allow a straightforward interpretation of results and recapitulate aspects of pathophysiology. For instance, dissecting the AD-related processes (e.g., neurotoxicity vs. synaptotoxicity) may be difficult with the common cell-based systems such as neuronal cell lines or primary neurons. To investigate and characterize the impact of reactive astrocytes on neuronal morphology in the context of AD-related cues, we modified an in vitro co-culture assay of primary mouse neurons and primary mouse astrocytes based on the so-called Banker "sandwich" co-culture assay. Here, we provide a simple and modular assay with fully differentiated primary mouse neurons to study the paracrine interactions between the neurons and the astrocytes in the co-culture setting. Readouts were obtained from both cell types in our assay. Astrocyte feeder cells were pre-exposed to neuroinflammatory conditions by means of Aβ42, Aβ40, or lipopolysaccharide (LPS). Non-cell autonomous toxic effects of reactive astrocytes on neurons were assessed using the Sholl analysis to evaluate the dendritic complexity, whereas synaptic puncta served as a readout of synaptotoxicity. Here, we show that astrocytes actively contribute to the phenotype of the primary neurons in an AD-specific context, emphasizing the role of different cell types in AD pathology. The cytokine expression pattern was significantly altered in the treated astrocytes. Of note, the impact of reactive astrocytes on neurons was highly dependent on the defined cell ratios. Our co-culture system is modular, of low cost, and allows us to probe aspects of neurodegeneration and neuroinflammation between the two major CNS cell types, neurons, and astrocytes, under well-defined experimental conditions. Our easy-to-follow protocol, including work-flow figures, may also provide a methodological outline to study the interactions of astrocytes and neurons in the context of other diseases in the future.


Genetic and epigenetic changes in mammary epithelial cells may mimic early events in carcinogenesis.
Studies of human mammary epithelial cells from healthy individuals are providing novel insights into how early epigenetic and genetic events affect genomic integrity and fuel carcinogenesis. Key epigenetic changes, such as the hypermethylation of the p16 (INK4a) promoter sequences, create a previously unappreciated preclonal phase of tumorigenesis in which a subpopulation of mammary epithelial cells are positioned for progression to malignancy (Romanov et al. , 2001, Nature , 409:633-637; Tlsty et al. , 2001, J. Mammary Gland Biol. Neoplasia , 6:235-243). These key changes precede the clonal outgrowth of premalignant lesions and occur frequently in healthy, disease-free women. Understanding more about these early events should provide novel molecular candidates for prevention and therapy of breast cancer that target the process instead of the consequences of genomic instability. This review will highlight some of the key alterations that have been studied in human mammary epithelial cells in culture and relate them to events observed in vivo and discussed in accompanying reviews in this volume.


Relationships Among Arsenic-Related Traits, Including Rice Grain Arsenic Concentration and Straighthead Resistance, as Revealed by Genome-Wide Association.
There is global concern that rice grains and foods can contain harmful amounts of arsenic (As), motivating breeders to produce cultivars that restrict As accumulation in grains to protect human health. Arsenic is also toxic to plants, with straighthead disorder (StHD), causing panicle sterility, being observed in rice. The genetic variation in StHD resistance suggests that plants have evolved mechanisms that reduce As toxicity, possibly via regulation of As uptake, transport, or detoxification/sequestration. Because these mechanisms could also underlie the wide (3- to 100-fold) differences in grain As concentration (grain-As) observed among diverse rice genotypes, it was hypothesized that some genes reduce both grain-As content and StHD susceptibility and may be detectable as co-located StDH and As quantitative trait loci (QTL). We used a machine-learning Bayesian network approach plus high-resolution genome-wide association study (GWAS) to identify QTL for grain-As and StHD resistance within the USDA Rice Minicore Collection (RMC). Arsenic enters roots through phosphorus (P) and silica (Si) transporters, As detoxification involves sulfur (S), and cell signaling to activate stress tolerance mechanisms is impacted by Si, calcium (Ca), and copper (Cu). Therefore, concentrations of Si, P, S, Ca, and Cu were included in this study to elucidate physiological mechanisms underlying grain-As and StHD QTL. Multiple QTL (from 9 to 33) were identified for each of the investigated As-associated traits. Although the QTL for StHD, Si, and grain-As did not overlap as heavily as our hypothesis predicted (4/33 StHD and 4/15 As QTL co-located), they do provide useful guidance to future research. Furthermore, these are the first StHD and Si QTL to be identified using high-density mapping, resulting in their being mapped to shorter, more precise genomic regions than previously reported QTL. The candidate genes identified provide guidance for future research, such as gene editing or mutation studies to further investigate the role of antioxidants and ROS scavenging to StHD resistance, as indicated by candidate genes around the commonly reported qStHD8-2 QTL. Other genes indicated for future study for improving grain-As and StHD include several multidrug and toxic compound extrusion (MATE) genes, F-box genes, and NIPs not documented to date to transport As.


Antioxidant effect of leaf extracts from Cressa cretica against oxidation process in soybean oil.
Phenolic compounds from Cressa cretica leaves were extracted using different solvents (water and ethanol 70%) during 3-24 h by immersion method and were determined according to the Folin-Ciocalteu method. The antioxidant activities of the extracts were investigated by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging, ferric reduction activity potential and total antioxidant capacity assays compared with synthetic antioxidant namely BHT. The results were shown that the most total phenol content was outcome of the ethanolic extract after 18 h (68.512 ± 0.36 mg gallic acid/g dry extract) extraction. The results of different assays for determination of antioxidant potential of the extracts as well as their EC50 values were indicated antioxidant activities in order: BHT>ethanolic extract>aqueous extract. Also, the antioxidant activities were enhanced with increasing of the extracts and BHT concentrations. The results of Peroxide value (PV) and Thiobarbituric acid (TBA) tests were revealed that the ethanolic extract with various concentrations (200-1000 ppm) were suppressed the oxidation of soybean oil. The highest oxidation inhibitor was belonged to 1000 ppm concentration of the ethanolic extract so that can be good alternative for BHT.


Immunohistochemical analysis of MUC5B apomucin expression in breast cancer and non-malignant breast tissues.
A deregulation of several MUC genes (MUC1, MUC2, MUC3, MUC5AC, and MUC6) was previously demonstrated in breast carcinomas. Considering that recently we found the "non-mammary" MUC5B mRNA in primary breast tumors (Berois et al. 2003), we undertook the present study to evaluate the expression profile of MUC5B protein product in breast tissues, using LUM5B-2 antisera raised against sequences within the non-glycosylated regions of this apomucin. Expression of MUC5B by breast cancer cells was confirmed by immunocytochemistry, in situ hybridization, and Western blot on MCF-7 cancer cells. Using an immunohistochemical procedure, MUC5B apomucin was detected in 34/42 (81%) primary breast tumors, in 13/14 (92.8%) samples of non-malignant breast diseases, in 8/19 (42.1%) samples of normal-appearing breast epithelia adjacent to cancer, and in 0/5 normal control breast samples. The staining pattern of MUC5B was very different when comparing breast cancer cells (cytoplasmic) and non-malignant breast cells (predominantly apical and in the secretory material). We analyzed MUC5B mRNA expression using RT-PCR in bone marrow aspirates from 22/42 patients with breast cancer to compare with MUC5B protein expression in the primary tumors. Good correlation was observed because the six MUC5B-positive bone marrow samples also displayed MUC5B expression in the tumor. Our results show, for the first time at the protein level, that MUC5B apomucin is upregulated in breast cancer. Its characterization could provide new insights about the glycobiology of breast cancer cells.


Magnetic resonance imaging and multi-detector computed tomography assessment of extracellular compartment in ischemic and non-ischemic myocardial pathologies.
Myocardial pathologies are major causes of morbidity and mortality worldwide. Early detection of loss of cellular integrity and expansion in extracellular volume (ECV) in myocardium is critical to initiate effective treatment. The three compartments in healthy myocardium are: intravascular (approximately 10% of tissue volume), interstitium (approximately 15%) and intracellular (approximately 75%). Myocardial cells, fibroblasts and vascular endothelial/smooth muscle cells represent intracellular compartment and the main proteins in the interstitium are types I/III collagens. Microscopic studies have shown that expansion of ECV is an important feature of diffuse physiologic fibrosis (e.g., aging and obesity) and pathologic fibrosis [heart failure, aortic valve disease, hypertrophic cardiomyopathy, myocarditis, dilated cardiomyopathy, amyloidosis, congenital heart disease, aortic stenosis, restrictive cardiomyopathy (hypereosinophilic and idiopathic types), arrythmogenic right ventricular dysplasia and hypertension]. This review addresses recent advances in measuring of ECV in ischemic and non-ischemic myocardial pathologies. Magnetic resonance imaging (MRI) has the ability to characterize tissue proton relaxation times (T1, T2, and T2*). Proton relaxation times reflect the physical and chemical environments of water protons in myocardium. Delayed contrast enhanced-MRI (DE-MRI) and multi-detector computed tomography (DE-MDCT) demonstrated hyper-enhanced infarct, hypo-enhanced microvascular obstruction zone and moderately enhanced peri-infarct zone, but are limited for visualizing diffuse fibrosis and patchy microinfarct despite the increase in ECV. ECV can be measured on equilibrium contrast enhanced MRI/MDCT and MRI longitudinal relaxation time mapping. Equilibrium contrast enhanced MRI/MDCT and MRI T1 mapping is currently used, but at a lower scale, as an alternative to invasive sub-endomyocardial biopsies to eliminate the need for anesthesia, coronary catheterization and possibility of tissue sampling error. Similar to delayed contrast enhancement, equilibrium contrast enhanced MRI/MDCT and T1 mapping is completely noninvasive and may play a specialized role in diagnosis of subclinical and other myocardial pathologies. DE-MRI and when T1-mapping demonstrated sub-epicardium, sub-endocardial and patchy mid-myocardial enhancement in myocarditis, Behcet's disease and sarcoidosis, respectively. Furthermore, recent studies showed that the combined technique of cine, T2-weighted and DE-MRI technique has high diagnostic accuracy for detecting myocarditis. When the tomographic techniques are coupled with myocardial perfusion and left ventricular function they can provide valuable information on the progression of myocardial pathologies and effectiveness of new therapies.


Effect of different levels of positive end-expiratory pressure on lung water content.
To compare the effects of 2-, 5-, and 10-cmH2O positive end-expiratory pressure (PEEP) on pulmonary extravascular water volume (PEWV), pulmonary blood volume (PBV), pulmonary dry weight (PDW), and distensibility, we separately ventilated perfused dogs' lungs in situ and produced pulmonary edema with oleic acid (0.06 ml/kg). Three groups were studied: I, PEEP, 5 cmH2O in both lung; II, PEEP, 2 cmH2O in one lung and 10 cmH2O in the other; and III, PEEP, same as II, but the chest was rotated to compensate for differences in heights. The PEWV and distensibility were less (P less than 0.05) in lungs exposed to 10-cmH2O than to either 2- or 5-cmH2O PEEP. After chest rotation, the difference between 10- and 2-cmH2O PEEP on PEWV was eliminated but that on distensibility was not. We conclude that 10-cmH2O PEEP 1) decreased water content because of lung volume-induced effects on intravascular hydrostatic pressure and 2) improved distensibility by recruitment of alveoli, irrespective of PEWV.


Information sharing and environmental policies.
Based on the assumption that in a standard eco-dumping model governments are uncertain about future product demand and allowing governments to obtain information from firms, we examine governments' and firms' incentives to share information. We show that when governments regulate polluting firms through emission standards, then governments and firms will reach an agreement concerning information sharing. The opposite holds when governments regulate pollution through emission taxes.


Supernumerary Nostril: Two Years Follow-Up.
Supernumerary nostril is a very rare congenital anomaly of the nose. Since the first patient reported by Lindsay in 1906, few number of patients were reported in the literature. Various types had been described with different surgical modalities for correction. It can be isolated or associated with other malformations such as facial cleft, esophageal atresia, and imperforate anus. Most of the patients are unilateral, but it may be bilateral. It may have a communication with a normal nasal cavity or not.In this study, the authors present a case of a 1-year-old male with a positive perinatal history of teratogen exposure had isolated supernumerary left nostril with communication to the nasal cavity.The authors will discuss different theories related to supernumerary nostril development, differences between published reports, the proposed surgical techniques, and the outcome of their treatment approach.


The role of the hypothalamic-pituitary-adrenal (HPA) axis in the regulation of blood pressure.
The role of the HPA axis in blood pressure regulation was examined in 6 normal male volunteers by comparing haemodynamic and hormonal effects of placebo, captopril, and dexamethasone given in random order for two days. The average 24-hour systolic and mean arterial pressures on placebo (135 +/- 6 and 93 +/- 2 mmHg respectively) were significantly higher than on captopril (118 +/- 1 and 85 +/- 1 mmHg respectively, p < 0.05) but there were no significant changes on dexamethasone compared with placebo (128 +/- 3 and 89 +/- 3 mmHg respectively). There were no differences in the average 24-hour diastolic blood pressures or heart rates, nor the day-night differences, night:day ratios or percentage changes in blood pressure and heart rate between treatments. Captopril significantly increased active plasma renin concentration, whilst dexamethasone decreased cortisol concentration. These results confirm the role of the renin-angiotensin system in the regulation of blood pressure in normal subjects but suggest that the HPA axis does not play a major role in determining ambulatory blood pressure or day-night variability in the short term.


Nitisinone Treatment Affects Biomarkers of Bone and Cartilage Remodelling in Alkaptonuria Patients.
Nitisinone has been approved for treatment of alkaptonuria (AKU). Non-invasive biomarkers of joint tissue remodelling could aid in understanding the molecular changes in AKU pathogenesis and how these can be affected by treatment. Serological and urinary biomarkers of type I collagen and II collagen in AKU were investigated in patients enrolled in the randomized SONIA 2 (NCT01916382) clinical study at baseline and yearly until the end of the study (Year 4). The trajectories of the biomarkers over time were observed. After treatment with nitisinone, the biomarkers of type I collagen remodelling increased at Year 1 (19% and 40% increase in CTX-I and PRO-C1, respectively), which was potentially reflected in the higher degree of mobility seen following treatment. The biomarkers of type II collagen remodelling decreased over time in the nitisinone group: C2M showed a 9.7% decline at Year 1, and levels then remained stable over the following visits; CTX-II showed a 26% decline at Year 3 and 4 in the nitisinone-treated patients. Nitisinone treatment induced changes in biomarkers of bone and cartilage remodelling. These biomarkers can aid patient management and deepen our knowledge of the molecular mechanisms of this rare disease.


High fat diet dysregulates microRNA-17-5p and triggers retinal inflammation: Role of endoplasmic-reticulum-stress.
To elucidate how high diet-induced endoplasmic reticulum-stress upregulates thioredoxin interacting protein expression in Müller cells leading to retinal inflammation. Male C57Bl/J mice were fed either normal diet or 60% high fat diet for 4-8 wk. During the 4 wk study, mice received phenyl-butyric acid (PBA); endoplasmic reticulum-stress inhibitor; for 2 wk. Insulin resistance was assessed by oral glucose tolerance. Effects of palmitate-bovine serum albumin (BSA) (400 μmol/L) were examined in retinal Müller glial cell line and primary Müller cells isolated from wild type and thioredoxin interacting protein knock-out mice. Expression of thioredoxin interacting protein, endoplasmic reticulum-stress markers, miR-17-5p mRNA, as well as nucleotide-binding oligomerization domain-like receptor protein (NLRP3) and IL1β protein was determined. High fat diet for 8 wk induced obesity and insulin resistance evident by increases in body weight and impaired glucose tolerance. By performing quantitative real-time polymerase chain reaction, we found that high fat diet triggered the expression of retinal endoplasmic reticulum-stress markers (P < 0.05). These effects were associated with increased thioredoxin interacting protein and decreased miR-17-5p expression, which were restored by inhibiting endoplasmic reticulum-stress with PBA (P < 0.05). In vitro, palmitate-BSA triggered endoplasmic reticulum-stress markers, which was accompanied with reduced miR-17-5p and induced thioredoxin interacting protein mRNA in retinal Müller glial cell line (P < 0.05). Palmitate upregulated NLRP3 and IL1β expression in primary Müller cells isolated from wild type. However, using primary Müller cells isolated from thioredoxin interacting protein knock-out mice abolished palmitate-mediated increase in NLRP3 and IL1β. Our work suggests that targeting endoplasmic reticulum-stress or thioredoxin interacting protein are potential therapeutic strategies for early intervention of obesity-induced retinal inflammation.


Role of endothelium and nitric oxide in the response of equine colonic arterial rings to vasoconstrictor agents.
To determine the in vitro contractile responses of equine colonic arteries to angiotensin II, histamine, serotonin, norepinephrine, prostaglandin F2 alpha, vasopressin, and a thromboxane-B2-analogue. The tension generated in colonic arterial rings placed in organ baths with oxygenated Tyrode's solution at 37 degrees C after exposure to the previously mentioned chemical agents was measured using force-transducers interfaced with a polygraph. Large colon arterial rings collected from eight horses. The rings were allowed to equilibrate for 45 minutes after applying 2 g tension. Bath solution was replaced and tension reapplied at 15-minute intervals. Cumulative-concentration-responses were determined for concentrations ranging from 10(-8) M to 10(-4) M on three vessel groups namely endothelium intact, endothelium denuded, and L-NAME treated. The maximal response for each vessel was considered as 100%; responses to lower concentrations were calculated as a percentage of the maximum. The EC50 value was determined for each concentration-response relationship of each agent. Vessels with denuded endothelium or those incubated with L-NAME had greater contractile responses. Angiotensin, histamine, serotonin, and norepinephrine produced greater maximal responses than the other agents. Endothelium denuded rings had lower EC50 values. Responses to norepinephrine and serotonin were affected less by denudation. Endothelium plays an important role in modulating responses of colonic arterial rings to contractile agents. Endothelium-derived vasodilators, other than nitric oxide, may modulate contractile responses of equine colonic arteries. Endothelial damage associated with colonic vovulus may be a major factor for sustained reduced perfusion after surgical correction.


Disease-causing mutations in the serpin antithrombin reveal a key domain critical for inhibiting protease activities.
Antithrombin mainly inhibits factor Xa and thrombin. The reactive center loop (RCL) is crucial for its interactions with its protease targets and is fully inserted into the A-sheet after its cleavage, causing translocation of the covalently linked protease to the opposite end of the A-sheet. Antithrombin variants with altered RCL hinge residues behave as substrates rather than inhibitors, resulting in stoichiometries of inhibition greater than one. Other antithrombin residues have been suggested to interfere with RCL insertion or the stability of the antithrombin-protease complex, but available crystal structures or mutagenesis studies have failed to identify such residues. Here, we characterized two mutations, S365L and I207T, present in individuals with type II antithrombin deficiency and identified a new antithrombin functional domain. S365L did not form stable complexes with thrombin or factor Xa, and the I207T/I207A variants inhibited both proteases with elevated stoichiometries of inhibition. Close proximity of Ile-207 and Ser-365 to the inserted RCL suggested that the preferred reaction of these mutants as protease substrates reflects an effect on the rate of the RCL insertion and protease translocation. However, both residues lie within the final docking site for the protease in the antithrombin-protease complex, supporting the idea that the enhanced substrate reactions may result from an increased dissociation of the final complexes. Our findings demonstrate that the distal end of the antithrombin A-sheet is crucial for the last steps of protease inhibition either by affecting the rate of RCL insertion or through critical interactions with proteases at the end of the A-sheet.


Contrast-enhanced MR Imaging versus Contrast-enhanced US: A Comparison in Glioblastoma Surgery by Using Intraoperative Fusion Imaging.
Purpose To compare contrast material enhancement of glioblastoma multiforme (GBM) with intraoperative contrast-enhanced ultrasonography (US) versus that with preoperative gadolinium-enhanced T1-weighted magnetic resonance (MR) imaging by using real-time fusion imaging. Materials and Methods Ten patients with GBM were retrospectively identified by using routinely collected, anonymized data. Navigated contrast-enhanced US was performed after intravenous administration of contrast material before tumor resection. All patients underwent tumor excision with navigated intraoperative US guidance with use of fusion imaging between real-time intraoperative US and preoperative MR imaging. With use of fusion imaging, glioblastoma contrast enhancement at contrast-enhanced US (regarding location, morphologic features, margins, dimensions, and pattern) was compared with that at gadolinium-enhanced T1-weighted MR imaging. Results Fusion imaging for virtual navigation enabled matching of real-time contrast-enhanced US scans to corresponding coplanar preoperative gadolinium-enhanced T1-weighted MR images in all cases, with a positional discrepancy of less than 2 mm. Contrast enhancement of gadolinium-enhanced T1-weighted MR imaging and contrast-enhanced US was superimposable in all cases with regard to location, margins, dimensions, and morphologic features. The qualitative analysis of contrast enhancement pattern demonstrated a similar distribution in contrast-enhanced US and gadolinium-enhanced T1-weighted MR imaging in nine patients: Seven lesions showed peripheral inhomogeneous ring enhancement, and two lesions showed a prevalent nodular pattern. In one patient, the contrast enhancement pattern differed between the two modalities: Contrast-enhanced US showed enhancement of the entire bulk of the tumor, whereas gadolinium-enhanced T1-weighted MR imaging demonstrated peripheral contrast enhancement. Conclusion Glioblastoma contrast enhancement with contrast-enhanced US is superimposable on that provided with preoperative gadolinium-enhanced T1-weighted MR imaging regarding location, margins, morphologic features, and dimensions, with a similar enhancement pattern in most cases. Thus, contrast-enhanced US is of potential use in the surgical management of GBM. © RSNA, 2017 Online supplemental material is available for this article.


A prognostic nomogram constructed for relapsed or refractory diffuse large B-cell lymphoma patients.
The clinical course of diffuse large B-cell lymphoma (DLBCL) is variable and there is a lack of prognostic markers and models for relapsed or refractory (r/r) DLBCL. Hence, we conducted this study to identify independent factors that can predict the survival rate of r/r DLBCL patients. A total of 416 r/r DLBCL patients who were pretreated with first-line anthracycline-based chemotherapy at the National Cancer Center in China between 2006 and 2016 were divided into the primary (n = 291) and validation (n = 125) cohorts. The effect of preclinical and clinical indicators on DLBCL survival rates of the two cohorts were evaluated by univariate and multivariate analyses. Factors showing good correlation with patient survival rates were used to construct a prognostic nomogram. Multivariate analysis of the primary cohort revealed five independent prognostic factors: lactate dehydrogenase level at diagnosis, response to front line treatment, progression/recurrence pattern, location, and invasion on progression/recurrence, which were then used to construct a nomogram. The nomogram was shown to have a C-index of 0.76 and AUC values of 0.81 and 0.80 for the primary and validation cohorts, respectively, suggesting good prognostic power. We further stratified the r/r DLBCL patients into four risk groups according to the newly developed nomogram. The prognostic nomogram constructed using the five identified clinical indicators can potentially be applied in the clinical setting to guide treatment decision.


Exploring risk for abuse of children with chronic conditions or disabilities--parent's perceptions of stressors and the role of professionals.
Children with chronic conditions or disabilities are at an increased risk for abuse. High level of parental stress has been identified as possible trigger for abuse, were a combination of several factors are of importance, as lack of social support and limited resources in the neighbourhood. Suggestions for preventive measures have merely focused on parenting strategies and targeted intervention programmes. So far, little attention has been paid on how the risk for abuse might relate to parent's perceptions of stressors and the role of professionals. The purpose of the current study was therefore to explore risk factors for abuse with focus on both parent-child and parent-professional relationships. Semi-structured in-depth interviews with 15 parents of children with chronic conditions or disability were collected and analysed according to qualitative content analysis. Three major themes were found that could be seen as risk factors for child abuse: (1) Emotional demands in precarious situations between parent and child. (2) Gradual shift in responsibility from professionals to parents concerning access to and co-ordination of service and support. (3) Emotionally closed environment between professionals and parents and taboo on talking about abuse. The gradual shift in responsibility had emotional implications, which could reinforce parental stress and thereby also indirect increase the risk of child abuse. The gradual shift in responsibility also seemed to generate an emotionally closed environment and reinforce the taboo on talking about abuse, which in turn hindered preventive measures. In the light of parent's perceptions of stressors and the role of professionals the findings indicate that abuse against children with chronic conditions or disability is not only a family matter, but also depending on qualities in service, professional support and social norms. The result pinpoints three challenges for preventive measures, all with emotional implications, parental strategies, organizational efforts and cultural awareness.


LINC01268 promotes epithelial-mesenchymal transition, invasion and metastasis of gastric cancer via the PI3K/Akt signaling pathway and targeting MARCKS.
Long non-coding RNAs (lncRNAs) with differential expression characteristics have been found to be closely related to the tumorigenesis and development of gastric cancer (GC), but their specific mechanisms and roles still need to be further elucidated. To investigate the expression of LINC01268 in GC and its mechanism of affecting GC progression. Real-time quantitative polymerase chain reaction was used to detect the expression of LINC01268 in GC tissues, cell lines and plasma. The Kaplan-Meier method was used to evaluate the value of LINC01268 in the prognostication of GC patients. An receiver operating characteristic curve was constructed to evaluate the value of LINC01268 in the diagnosis of GC. Transwell migration and invasion assays and wound healing assays were used to confirm the effect of LINC01268 on the invasion and migration of GC cells. The regulatory relationship between LINC01268 and myristoylated alanine rich protein kinase C substrate (MARCKS), the PI3K/Akt signaling pathway, and the epithelial-mesenchymal transition (EMT) process in GC was demonstrated by western blot analysis. The expression of LINC01268 was increased in GC tissues and cell lines. The expression level of LINC01268 was significantly correlated with lymph node metastasis, TNM stage, and tumor differentiation in patients with GC. Over-expression of LINC01268 indicated a poor prognosis for patients with GC, and it had a certain auxiliary diagnostic value for GC. In vitro functional experiments proved that the abnormal expression of LINC01268 further activated the PI3K/Akt signaling pathway and promoted EMT by targeting and regulating MARCKS and ultimately promoted the invasion and metastasis of GC. This study elucidates that LINC01268 in GC may be an oncogene that further activates the PI3K/Akt signaling pathway and EMT by targeting and regulating MARCKS, and ultimately promotes the invasion and metastasis of GC. LINC01268 may be a potential effective target for the treatment of GC.


Cytokinin activity of N6-benzyladenine derivatives assayed by interaction with the receptors in planta, in vitro, and in silico.
Biological effects of hormones in both plants and animals are based on high-affinity interaction with cognate receptors resulting in their activation. The signal of cytokinins, classical plant hormones, is perceived in Arabidopsis by three homologous membrane receptors: AHK2, AHK3, and CRE1/AHK4. To study the cytokinin-receptor interaction, we used 25 derivatives of potent cytokinin N6-benzyladenine (BA) with substituents in the purine heterocycle and/or in the side chain. The study was focused primarily on individual cytokinin receptors from Arabidopsis. The main in planta assay system was based on Arabidopsis double mutants retaining only one isoform of cytokinin receptors and harboring cytokinin-sensitive reporter gene. Classical cytokinin biotest with Amaranthus seedlings was used as an additional biotest. In parallel, the binding of ligands to individual cytokinin receptors was assessed in the in vitro test system. Quantitative comparison of results of different assays confirmed the partial similarity of ligand-binding properties of receptor isoforms. Substituents at positions 8 and 9 of adenine moiety, elongated linker up to 4 methylene units, and replacement of N6 by sulfur or oxygen have resulted in the suppression of cytokinin activity of the derivative toward all receptors. Introduction of a halogen into position 2 of adenine moiety, on the contrary, often increased the ligand activity, especially toward AHK3. Features both common and distinctive of cytokinin receptors in Arabidopsis and Amaranthus were revealed, highlighting species specificity of the cytokinin perception apparatus. Correlations between the extent to which a compound binds to a receptor in vitro and its ability to activate the same receptor in planta were evaluated for each AHK protein. Interaction patterns between individual receptors and ligands were rationalized by structure analysis and molecular docking in sensory modules of AHK receptors. The best correlation between docking scores and specific binding was observed for AHK3. In addition, receptor-specific ligands have been discovered with unique properties to predominantly activate or block distinct cytokinin receptors. These ligands are promising for practical application and as molecular tools in the study of the cytokinin perception by plant cells.


The new colposcopic terminology.
A generally acceptable scheme for the classification of colposcopic findings is critical for the more widespread acceptance of this valuable diagnostic tool. Older schemes have tended to confuse students. The scheme described in this article sets appearances against an order of events in time and place based on the biological life history of the organ. This amounts to the history of exposed columnar epithelium as it changes, by a typical or atypical process, into squamous epithelium to produce, respectively, a typical or atypical transformation zone. The former produces a homogeneous appearance, the latter a heterogeneous appearance requiring further subdivision by the use of the categories of white epithelium, blood vessel abnormalities such as punctation and mosaic and other atypical patterns, and leukoplakia. Not all of these abnormal appearances are associated with greatly altered histological appearances and, therefore, a further system of their grading is introduced. Other categories include appearances associated with overt cancer, with miscellaneous noncancerous conditions and with states in which the complete transformation zone is not viewed and, therefore, the colposcopic findings are not satisfactory. Finally, the present scheme is integrated as far as possible with older schemes.


Gender, aging and power in sub-Saharan Africa: Challenges and puzzles.
By 2020, the absolute numbers of elderly in Africa will increase dramatically. A majority of these will be women. While we know much about the powers and authority of male elders within formal kin- and age-based structures, we know little about the lived experience of aging in Africa today, and even less about the formal and informal roles of elderly women. These ethnographic descriptions of gendered aging experiences in seven African societies examine the following little explored topics: indigenous conceptions of gender and aging; the ambiguities in women's social positions; changing family, household and marital relationships, which affect experiences of old age; older persons' access to and control over material, human and suprahuman resources; and the interplay of indigenous and modern forces. They point to future research challenges that will include understanding indigenous notions of gender and aging, and power and personhood, as they relate to personal experiences and to the ability of older women and men to assure their security in contexts of rapid social change.


Investigating the mechanism of hysteresis effect in graphene electrical field device fabricated on SiO₂ substrates using Raman spectroscopy.
The hysteresis effect is a common problem in graphene field-effect transistors (FETs). Usually, the external doping to graphene is considered to be responsible for the hysteresis behavior, but is not yet clearly understood. By monitoring the doping of graphene and the hysteresis in graphene FETs under different atmospheres using in situ Raman spectroscopy, it is confirmed that the electrochemical doping of O(2) /H(2) O redox couple to graphene is responsible for the hysteresis effect. In addition, Raman spectra of graphene on SiO(2) substrate show stronger doping than that suspended, which indicates that SiO(2) substrate plays an important role in the doping of graphene. Herein it is proposed that the doping species (H(2) O and O(2) ) are bounded at the interface of graphene/SiO(2) substrate by hydrogen-bonds with the silanol groups on SiO(2) substrate. The dynamic equilibrium process of the charge-transfer between H(2) O/O(2) redox couple and graphene under electrical field modulation is carefully analyzed using Marcus-Gerischer theory. This work provides a clear view to the mechanism of the hysteresis effect, and is of benefit to a reliable design to suppress the hysteresis in graphene FETs.


Modeling intrahippocampal effects of anterior hippocampal hyperactivity relevant to schizophrenia using chemogenetic excitation of long axis-projecting mossy cells in the mouse dentate gyrus.
The anterior hippocampus of individuals with early psychosis or schizophrenia is hyperactive, as is the ventral hippocampus in many rodent models for schizophrenia risk. Mossy cells (MCs) of the ventral dentate gyrus (DG) densely project in the hippocampal long axis, targeting both dorsal DG granule cells and inhibitory interneurons. Mossy cells are responsive to stimulation throughout hippocampal subfields, and thus may be suited to detect hyperactivity in areas where it originates such as CA1. Here we tested the hypothesis that hyperactivation of ventral MCs activates dorsal DG granule cells to influence dorsal hippocampal function. In CD-1 mice, we targeted dorsal DG-projecting ventral MCs using an adeno-associated virus intersectional strategy. In vivo fiber photometry recording of ventral MCs was performed during exploratory behaviors. We used excitatory chemogenetic constructs to test the effects of ventral MC hyperactivation on long-term spatial memory during an object location memory task. Photometry revealed ventral MCs were activated during exploratory rearing. Ventral MCs made functional monosynaptic inputs to dorsal DG granule cells, and chemogenetic activation of ventral MCs modestly increased activity of dorsal DG granule cells measured by c-Fos. Finally, chemogenetic activation of ventral MCs during the training phase of an object location memory task impaired test performance 24 hours later, without effects on locomotion or object exploration. These data suggest that ventral MC activation can directly excite dorsal granule cells and interfere with dorsal DG function, supporting future study of their in vivo activity in animal models for schizophrenia featuring ventral hyperactivity.


Using a divider nudge in supermarket shopping trolleys to increase fruit and vegetable purchases: A feasibility study using an intervention design.
Fruit and vegetable (F&V) consumption is associated with a reduced risk of developing obesity and chronic diseases: however, only one in 16 Australian adults consume F&Vs at the recommended two servings of fruit and five servings of vegetables per day. What and how much people eat is influenced by their social and physical environments. Supermarkets are a key setting influencing food purchases, and as such, they can shape consumption patterns of F&Vs. Implementing effective strategies to increase F&V intake is crucial. The objective of this research was to test the feasibility of covertly modifying shopper purchasing behaviour to purchase more F&Vs using a visual divider nudge message (prompts) covering the entire base of shopping trolleys. Placards provided a visual representation of the recommended proportion of the trolley base that should be allocated to fruits and vegetables (implied social norm). Applying an intervention research design, 30 out of ~100 trolleys were fitted with the placards and shopper purchases were measured by collecting receipts to measure the weight (kg), total spending and F&V specific spending (Australian dollars) for intervention versus control trolleys for one weekend day only. We also conducted a short intercept survey that was administered independently from the research study day on non-trial shoppers. Shoppers who selected trolleys with the divider nudge placards (n = 102) purchased equal weight of F&Vs (Intervention: mean = 6.25 kg, SD = 5.60 kg, 95% CI = 5.14 kg, 7.35 kg, vs. Control: mean 6.03 kg, SD = 5.17 kg, 95% CI = 5.01 kg, 7.04 kg, p = 0.768) and spent equal amounts on F&Vs compared to shoppers in the control group (n = 102) (Intervention: mean = $41.46, SD = $36.68, 95% CI = $34.25, $48.66, vs. Control: mean $39.85, SD = $33.30, 95% CI = $33.34, $46.39, p = 0.744). There was no difference in the total spending between groups (Intervention: mean = $135.99, SD = $90.10, 95% CI = $118.29, $153.68, vs. Control: mean $155.68, SD = $96.46, 95% CI = $136.73, $174.63, p = 0.133). The divider nudge placard did not lead to any difference in shoppers' purchases of F&Vs. However, this study demonstrates the feasibility of testing a cheap, simple and easy supermarket nutrition intervention. Larger studies are required to elucidate and confirm these findings over the longer term.


Next generation fixed dose combination pharmacotherapies for treating HIV.
Treatment options for patients with HIV-1 infection have grown over the past two decades to include multiple fixed-dose combination pharmacotherapies that have greatly simplified administration of antiretroviral therapy (ART) for both patients and providers. Effective virologic control can often be achieved with once-daily use of a single-tablet regimen. Over the past three years, ART drug development has focused on the next generation of fixed-dose combinations for initial and maintenance therapy with improved efficacy, safety and tolerability. This review covers pre-clinical and clinical data searched through PubMed and presented at major conferences through November 2017. Currently available single-tablet regimens have clinical limitations related to adverse event profiles, drug-drug and drug-food interactions and variable barriers to resistance. Anticipated advances in ART fixed-dose combinations promise combinations of current multiple tablet regimens into single tablets, as well as combinations with novel drugs with improved safety and tolerability. The traditional dogma of effective ART containing at least three active antiretroviral drugs is being challenged by promising data to support efficacy of certain regimens containing two drugs. Implementation of next generation ART will bring to light issues of clinical preference and cost-effectiveness as patents of existing drugs expire and more generic formulations become available.


Analysis of Peripapillary and Macular Choroidal Thickness in Eyes with Pseudoexfoliative Glaucoma and Fellow Eyes.
To compare differences in peripapillary and macular choroidal thickness in pseudoexfoliative glaucoma (PXG) eyes, nonexfoliative fellow eyes, and normal eyes. This case-control study included 37 PXG patients (group A: 37 PXG eyes; group B: 37 nonexfoliative fellow eyes) and 37 sex-, age-, and axial length-matched healthy volunteer eyes (group C). Peripapillary and macular choroidal thickness and volume were measured in all subjects via enhanced-depth imaging-optical coherence tomography. The average peripapillary (AP) choroidal thickness was (130.10 ± 46.14) μm, (131.43 ± 46.00) μm, and (147.89 ± 53.32) μm; average macular (AM) choroidal thickness was (191.72 ± 68.07) μm, (204.62 ± 69.54) μm, and (215.10 ± 45.40) μm; and average volume was (0.59 ± 0.21) μm3, (0.63 ± 0.21) μm3, and (0.65 ± 0.14) μm3 in groups A, B, and C, respectively. NIP choroidal thickness was significantly lower in groups A and B than in group C (P < 0.05). TIM and TOM choroidal thickness and volume were significantly lower in group A than in group C (P < 0.05). NIM, SIM, NOM, IOM, AM choroidal thickness and volume, and CSM choroidal thickness were significantly lower in group A than in group B (P < 0.05). CSM, TIM, and TOM in group A and TIM, TOM choroidal thickness, and volume in group B were significantly lower than in group C (P < 0.05). NIP choroidal thickness in PXG eyes and nonexfoliative fellow eyes and temporal macular choroidal thickness in PXG eyes were significantly lower than in normal eyes. Macular choroidal thickness (except in temporal regions) was significantly lower in PXG eyes than in nonexfoliative fellow eyes. Changes in peripapillary and macula choroidal thickness further elucidate the choroid's role in PXG development and progression.


Pioglitazone ameliorates doxorubicin-induced hypothyroidism and cardiotoxicity in rat models.
OBJECTIVE: The anticancer drug doxorubicin (DOX) is effective but is associated with complications such as hypothyroidism and cardiotoxicity. Pioglitazone (PIO), which is used to treat diabetes mellitus, has shown potential for treating hypothyroidism and cardiac dysfunction. Therefore, this study explores whether PIO can also ameliorate DOX-induced hypothyroidism and cardiotoxicity. MATERIALS AND METHODS: Forty female Wistar rats were separated into control and three treated groups (DOX, PIO, and DOX+PIO), and their blood samples were examined for the thyroid hormones, including thyroid-stimulating hormone (TSH), thyroxine in total and free forms (T4 and FT4, respectively), and triiodothyronine in total and free forms (T3 and FT3, respectively), and the cardiotoxicity biomarkers [troponin I, creatine kinase (CK), and creatine kinase-myocardial band (CK-MB)]. RESULTS: The control and PIO groups did not exhibit significant alterations in any of the examined hormones and markers. In contrast, in the DOX group, T4, FT4, T3, and FT3 levels decreased significantly, whereas troponin I, CK, and CK-MB levels increased significantly, but no significant changes were detected in TSH levels. PIO co-treatment ameliorated these effects of DOX significantly in FT4, FT3, and troponin I. CONCLUSIONS: PIO may provide protection against hypothyroidism and cardiotoxicity caused by DOX treatment, by significant reversal of FT4, FT3, and troponin I levels. https://www.europeanreview.org/wp/wp-content/uploads/Graphical-abstract1.jpg.


CT diagnosis of segmental pulmonary artery embolus.
A patient with pathologically proven pulmonary embolism that was diagnosed on computed tomography (CT) is presented. The clot in an artery with a diameter less than 1 cm was clearly demonstrated as was an associated pulmonary infarct, which appeared hyperdense on contrast enhanced scan. The potential of CT to noninvasively demonstrate pulmonary embolism, particularly in smaller arteries, is discussed.


Smoking Is Not an Independent Risk Factor for Increased Opioid Consumption in Patients Being Treated for Ankle Fractures.
Using narcotics for pain management is an integral part of orthopaedic surgery, especially after traumatic injuries such as ankle fractures. Although narcotics are often necessary for adequate pain control, prolonged duration of opioid treatment is becoming more common, and the detrimental effects of opioid use are well known. To treat this epidemic, we need to better understand the factors that put patients at risk for increased narcotic requirements and potential opioid misuse after orthopaedic injuries. The purpose of this study was to retrospectively compare opioid use among ankle fracture patients in smokers versus nonsmokers to better elucidate whether tobacco use is a risk factor for increased opioid consumption. A retrospective review was conducted for all patients who presented with an ankle fracture (Weber A to C) to any of 8 institutions in a large mid-Atlantic regional hospital system and subsequently underwent surgery between the November 2013 and January 2017. Exclusion criteria included patients age <18 years, a diagnosis of a pilon fracture, polytrauma, history of substance abuse, chronic pain syndromes, and osteoporosis. This yielded a total of 130 patients (96 nonsmokers and 34 smokers). A repeated-measures analysis of variance was conducted to compare opioid consumption between the 2 groups. Model analysis demonstrated no difference between the 2 groups (p = .782). There was no difference in opioid consumption at 1 month (p = .838), 2 months (p = .569), or 3 months (p = .656) between smokers and nonsmokers. Our study revealed no significant difference in opioid consumption among smokers compared with nonsmokers in patients treated for ankle fractures.


Case-control research. Temporal precedence and other problems of the exposure-disease relationship.
We assessed the principle of temporal precedence in recent case-control studies demonstrating the alleged associations between tampon use and toxic shock syndrome and between aspirin use and Reye's syndrome. For both relationships, we considered four components of the exposure-disease association, including: (1) establishing that the agent preceded the disease, (2) selecting an index time, (3) defining criteria for classifying a patient as "exposed," and (4) avoiding the bias that occurs when use of the etiologic agent was influenced by an early manifestation of the disease. The problems can be minimized by interviewing patients early during the course of their illness and by improving strategies for data analysis.


Neutrophil extracellular traps in ischemic stroke thrombi.
Neutrophil extracellular traps (NETs) have been shown to promote thrombus formation. Little is known about the exact composition of thrombi that cause ischemic stroke. In particular, no information is yet available on the presence of NETs in cerebral occlusions. Such information is, however, essential to improve current thrombolytic therapy with tissue plasminogen activator (t-PA). This study aimed at investigating the presence of neutrophils and more specifically NETs in ischemic stroke thrombi. Sixty-eight thrombi retrieved from ischemic stroke patients undergoing endovascular treatment were characterized by immunostaining using neutrophil markers (CD66b and neutrophil elastase) and NET markers (citrullinated histone H3 [H3Cit] and extracellular DNA). Neutrophils and NETs were quantified. In addition, extracellular DNA was targeted by performing ex vivo lysis of retrieved thrombi with DNase 1 and t-PA. Neutrophils were detected extensively throughout all thrombi. H3Cit, a hallmark of NETs, was observed in almost all thrombi. H3Cit-positive area varied up to 13.45% of total thrombus area. Colocalization of H3Cit with extracellular DNA released from neutrophils confirmed the specific presence of NETs. H3Cit was more abundant in thrombi of cardioembolic origin compared to other etiologies. Older thrombi contained significantly more neutrophils and H3Cit compared to fresh thrombi. Interestingly, ex vivo lysis of patient thrombi was more successful when adding DNase 1 to standard t-PA. Neutrophils and NETs form important constituents of cerebral thrombi. Targeting of NETs with DNase 1 might have prothrombolytic potential in treatment of acute ischemic stroke. Ann Neurol 2017;82:223-232.


New aspects of catalytic intramolecular C-H amination: unexpected formation of a seven-membered ring in nitrogen-containing systems.
The first example of the formation of a seven-membered ring by way of intramolecular-catalyzed amination of saturated C-H bonds is reported (Du Bois reaction). The influence of various structural parameters was studied, and it was shown that the unexpected regioselectivity observed in nitrogen-containing systems could be rationalized by conformational factors. These results open the way to innovative strategies for the general synthesis of polyfunctionalized piperidines.


Charting and Handling Therapeutic Drug Monitoring Results: How they Differ From Most Laboratory Results.
Therapeutic drug monitoring (TDM) results are a biological measure of the ability of the patient to clear a drug as long as the patient is adhering to the prescribed dosage. If the patient is not adherent, then TDM can provide evidence of such nonadherence. To properly interpret TDM results, several variables must be considered. First and foremost is the dose the patient is supposed to be taking. The second variable is whether the patient should have achieved a steady-state condition, which means that the amount of drug the patient is taking in a day is equal to the amount the patient is eliminating in the same day so that the level will not change unless changes occur in either the dosing rate or the ability of the patient to clear the drug. For drugs with a half-life of 24 hours, steady state typically occurs when the patient has been taking the same dose for 5 to 7 days (ie, 5 x the half-life). The third variable is the time between when the last dose was taken and the sample was obtained. Ideally, for most drugs, that desired interval is 10 to 12 hours after the last dose was taken assuming the patient is taking the dose daily as different rules apply for depot medications. There are 2 reasons for this stipulation. First, this interval will best reflect the average concentration that the patient achieves over the daily dosing interval on that drug. Second, for most orally administered drugs, the value that is obtained 10 to 12 hours after the last dose will not be influenced by differences in the absorption of the drug. In contrast, if the sample is obtained during the absorption phase, the plasma concentration can be both much higher and less predictable than is the case when absorption is complete. Therefore, these 3 variables must be included when recording TDM results in the chart. As explained in this column, the TDM result without this information is not optimally interpretable. Parenthetically, TDM results do not need to be repeated (unless specific concerns arise as explained in this column), because these results are a measure of the ability of the patient to clear the drug. For depot medications (eg, antipsychotics), the results should be obtained at the end of the dosing cycle just before the administration of the drug and after the patient has ideally received 4 to 5 injections at equal intervals (eg, monthly) to be near steady-state. Parenthetically, TDM at the appropriate fixed interval (usually monthly) with depot medications is much less sensitive to timing issues than TDM of orally administered drugs. This column explains the rationale for obtaining TDM results, principally focusing on orally administered drugs and how to optimally chart and disseminate these results.


Epoxy-rich Fe Single Atom Sites Boost Oxygen Reduction Electrocatalysis.
Electrocatalysts for highly efficient oxygen reduction reaction (ORR) are crucial for energy conversion and storage devices. Single-atom catalysts with maximized metal utilization and altered electronic structure are the most promising alternatives to replace current benchmark precious metals. However, the atomic level understanding of the functional role for each species at the anchoring sites is still unclear and poorly elucidated. Herein, we report Fe single atom catalysts with the sulfur and oxygen functional groups near the atomically dispersed metal centers (Fe1/NSOC) for highly efficient ORR. The Fe1/NSOC delivers a half-wave potential of 0.92 V vs. RHE, which is much better than those of commercial Pt/C (0.88 V), Fe single atoms on N-doped carbon (Fe1/NC, 0.89 V) and most reported nonprecious metal catalysts. The spectroscopic measurements reveal that the presence of sulfur group induces the formation of epoxy groups near the FeN4S2 centers, which not only modulate the electronic structure of Fe single atoms but also participate the catalytic process to improve the kinetics. The density functional theory calculations demonstrate the existence of sulfur and epoxy group engineer the charges of Fe reactive center and facilitate the reductive release of OH* (rate-limiting step), thus boosting the overall oxygen reduction efficiency.


Interacting factors and cellular localization of SR protein-specific kinase Dsk1.
Schizosaccharomyces pombe Dsk1 is an SR protein-specific kinase (SRPK), whose homologs have been identified in every eukaryotic organism examined. Although discovered as a mitotic regulator with protein kinase activity toward SR splicing factors, it remains largely unknown about what and how Dsk1 contributes to cell cycle and pre-mRNA splicing. In this study, we investigated the Dsk1 function by determining interacting factors and cellular localization of the kinase. Consistent with its reported functions, we found that pre-mRNA processing and cell cycle factors are prominent among the proteins co-purified with Dsk1. The identification of these factors led us to find Rsd1 as a novel Dsk1 substrate, as well as the involvement of Dsk1 in cellular distribution of poly(A)(+) RNA. In agreement with its role in nuclear events, we also found that Dsk1 is mainly localized in the nucleus during G(2) phase and at mitosis. Furthermore, we revealed the oscillation of Dsk1 protein in a cell cycle-dependent manner. This paper marks the first comprehensive analysis of in vivo Dsk1-associated proteins in fission yeast. Our results reflect the conserved role of SRPK family in eukaryotic organisms, and provide information about how Dsk1 functions in pre-mRNA processing and cell-division cycle.


Exotic QTL improve grain quality in the tri-parental wheat population SW84.
Genetic diversity of cultivated wheat was markedly reduced, first, during domestication and, second, since the onset of modern elite breeding. There is an increasing demand for utilizing genetic resources to increase genetic diversity and, simultaneously, to improve agronomic performance of cultivated wheat. To locate favorable effects of exotic wheat alleles, we developed the tri-parental wheat population SW84. The population was derived from crossing the hexaploid spring wheat cultivars Triso and Devon with one synthetic exotic donor accession, Syn084L, followed by two rounds of backcrossing and three rounds of selfing. SW84 consists of 359 BC2F4 lines, split into two families, D84 (Devon*Syn084L) and T84 (Triso*Syn084L). As a case study, grain quality of SW84 was studied in replicated field trials. Transgressive segregation was observed for all studied grain quality traits by evaluating SW84 for two years at two locations under low and high nitrogen supply. Subsequently, a genome-wide association study (GWAS) was carried out based on genomic data derived from a 90k Infinium iSELECT single nucleotide polymorphism (SNP) array. In total, GWAS yielded 37 marker-trait associations, summarized to 16 quantitative trait loci (QTL). These SNPs indicate genetic regulators of grain protein content, grain hardness, sedimentation value and sedimentation ratio. The majority of exotic QTL alleles (75%) exerted favorable effects, increasing grain protein content and sedimentation value in ten and two cases, respectively. For instance, two exotic QTL alleles were associated with a substantial increase of grain protein content and sedimentation value by 1.09% and 7.31 ml, respectively. This finding confirms the potential of exotic germplasm to improve grain quality in cultivated wheat. So far, the molecular nature of most of the detected QTL is unknown. However, two QTL correspond to known genes controlling grain quality: The major QTL on chromosome 6B, increasing grain protein content by 0.70%, on average, co-localizes with the NAM-B1 gene, known to control grain protein content as well as iron and zinc content. Likewise, the major QTL on chromosome 5D, reducing grain hardness by 8.98%, on average, co-localizes with the gene for puroindoline b (Pinb-D1) at the Ha locus. In total, 13 QTL were detected across families, whereas one and three QTL were exclusively detected in families D84 and T84, respectively. Likewise, ten QTL were detected across nitrogen treatments, whereas one and five QTL were exclusively detected under low and high N treatments, respectively. Our data indicate that most effects in SW84 act across families and N levels. Merging of data from two families or two N treatments may, thus, be considered in association studies to increase sample size and, as a result, QTL detection power. Our study serves as a model how favorable exotic QTL alleles can be located in exotic germplasm of wheat. In future, the localized favorable exotic QTL alleles will be utilized in wheat breeding programs to simultaneously improve grain quality and selectively expand genetic diversity of the elite wheat gene pool.


Effects of cimetidine, H2 receptor antagonist, on follicular and luteal development in the mice.
The effects of various doses, duration and frequency of cimetidine treatment on vaginal cycle, ovulation, ovarian histology and follicular kinetics were investigated. In addition, studies were performed to assess the reversibility in ovarian functions following withdrawal of cimetidine treatment. A statistically significant (P < 0.05) decline in ovarian and uterine weights, but not in body and adrenal weights, were observed in females treated with cimetidine four times daily for long-term as compared with the controls. Ovarian and uterine weights recovered to pretreatment levels following withdrawal of the treatment. Cimetidine treatment caused irregularities in the reproductive cyclicity of mice. Cimetidine treatment causes adverse effect on ovarian function depending on the time, duration and frequency of treatment. Treatment for only one day at proestrus induced significant (P < 0.05) decline in ovulatory efficiency. However, administration of cimetidine, four times daily, for either 6 or 14 days caused ovarian dysfunction. The treatment depressed the number of healthy preantral and antral follicles as well as number of healthy corpus luteum (CL) in the ovary. Absence of healthy CL, presence of several atretic late antral follicles and reproductive acyclicity provide evidence for the failure of ovulation in mice treated with cimetidine four times per day for 14 days. Moreover, when cimetidine was administered twice daily, the ovaries also showed newly formed CL. The results, thus, suggest that the effects of cimetidine on ovulation are dependent on dose, duration and frequency of treatment. Blockage of ovulation in long-term cimetidine treated mice could be due to its influence on follicular atresia. Ovulation had occurred in these females after withdrawal of cimetidine treatment.


FcgammaRIIa, IIIa and IIIb gene polymorphisms in Behçet's disease: do they have any clinical implications?
Behçet's disease (BD) is a unique systemic vasculitis involving both arteries and veins of all sizes. Since Fcgamma receptors (FcgammaR) are important in mediating various immune effector functions, FcgammaR gene polymorphisms may affect the susceptibility to systemic inflammatory diseases such as BD. The aim of this study was to show the distribution of FcgammaRIIa, IIIa ve IIIb receptor gene polymorphisms in BD, and to investigate possible genotype-phenotype relationships. In this cross-sectional study, FcgammaRIIa (H/H131, H/R131, R/R131), IIIa (F/F158, F/V158, V/V158), and IIIb (NA1/NA1, NA1/NA2, and NA2/NA2) receptor gene polymorphisms were investigated in 216 unrelated Turkish BD patients (M/F: 130/86) and in 241 healthy subjects, using an allele-specific polymerase chain reaction. The FcgammaRIIa R/R131 (p=0.019) and FcgammaRIIIa F/F158 genotypes (p=0.001) were found to be significantly more frequent in BD compared with healthy controls, whereas the FcgammaRIIIb genotypes were not (p=0.108). Allele analysis showed that the FcgammaRIIIa 158 (p=0.001) and FcgammaRIIIb NA2 (p=0.016) alleles were more frequent in BD than in healthy controls. In BD patients the FcgammaRIIIa V/V158 genotype was significantly associated with the presence of arthritis (p=0.002) and with an earlier disease onset (p=0.008), while the FcgammaRIIIb NA2/NA2 genotype was significantly associated with disease severity (p=0.02), vascular involvement (p=0.014), and pathergy positivity (p=0.02). We found that the genotype frequencies and allelic distributions of the FcgammaRIIa, FcgammaRIIIa and FcgammaRIIIb gene polymorphisms were significantly different between BD patients and healthy controls. In addition, certain FcgammaRIIIa and FcgammaRIIIb gene polymorphisms appear to be associated with an early disease onset, disease severity, the presence of arthritis, and vascular involvement in BD.


The FDG Pattern of Autonomously Functioning Thyroid Nodules Correlates With Thyroid-Stimulating Hormone and Histopathology.
Whereas the 18 F-FDG PET/CT pattern of malignant thyroid neoplasia is known, the glucose uptake of autonomously functioning thyroid nodules (AFTNs) has not been fully investigated. We aimed to analyze the FDG uptake of AFTNs and its correlation with clinical, laboratory, ultrasonography, and histological features. We retrospectively analyzed the records of all AFTN patients from 2 centers, in which an 18 F-FDG PET/CT, thyroid function tests, and neck ultrasound were available within a 3-month window from the thyroid scintigraphy. Forty-five patients (35 female patients; median age, 65 years) were retrospectively included. The presence of FDG uptake was compared with the laboratory data and, whenever available, the histology. Over a 36-month follow-up, 20 patients underwent surgery; 4 cancers, 10 follicular adenomas, and 6 follicular hyperplasias were found. Twenty-two AFTNs (48.9%) were FDG-positive, whereas the remaining 23 (51.1%) were not. Thyroid-stimulating hormone (TSH) was significantly lower in FDG-positive AFTNs than in negative ones (0.055 [0.02-0.42] vs 0.65 [0.2-0.96] mIU/L, P = 0.0018). On multivariate analysis, only TSH was independently associated with FDG uptake ( P = 0.008). On receiver operating characteristic curve analysis, TSH <0.08 mIU/L indicated FDG-positive AFTNs, with 64% sensitivity, 87% specificity, 4.88 positive likelihood ratio, and 0.42 negative likelihood ratio. In histologically proven benign lesions, TSH was significantly lower in follicular adenomas than in follicular hyperplasias ( P < 0.001). Patients with cancer had TSH in the low-normal range. Autonomously functioning thyroid nodules show heterogeneous uptake FDG pattern, which depends on histopathology and thyroid function. Particularly, follicular adenomas tend to display increased glucose uptake and suppressed TSH.


A systematic review of modeling approaches in green supply chain optimization.
Over the past decade, the significance of optimizing green supply chain management (GSCM) has gained unprecedented attention from both scholars and industry professionals. This surge in interest has led researchers to employ diverse modeling approaches in the pursuit of enhancing green supply chain networks. In this systematic review, we analyze 159 recent GSCM optimization papers published from 2017 to 2022 and identify the recent trends in mathematical modeling, multi-objective optimization, and the modeling/solver tools utilized. We find that the primary green focus is on minimizing carbon emissions (n = 44), reflecting the increasing concern for environmental sustainability. Among the modeling approaches employed, mixed-integer linear programming has emerged as the most popular choice (n = 51), followed by game theory-based modeling (n = 30). When it comes to multiobjective optimization, the ε-constraint approach is the most widely used. Evolutionary algorithms have emerged as the dominant meta-heuristic optimization approach. Additionally, the widely utilized solver in this domain is CPLEX with the most popular modeling/solver combination being GAMS/CPLEX. Moreover, the Journal of Cleaner Production was the leading outlet for research in this domain (n = 35). In addition to these findings, this study also discusses some other research trends and future research directions. Finally, we discuss the theoretical, managerial, and policy implications of this study. By providing GSCM researchers and practitioners with the latest trends in GSCM optimization approaches, this study contributes to the further advancement of the field.


Physiological Markers of Motor Inhibition during Human Behavior.
Transcranial magnetic stimulation (TMS) studies in humans have shown that many behaviors engage processes that suppress excitability within the corticospinal tract. Inhibition of the motor output pathway has been extensively studied in the context of action stopping, where a planned movement needs to be abruptly aborted. Recent TMS work has also revealed markers of motor inhibition during the preparation of movement. Here, we review the evidence for motor inhibition during action stopping and action preparation, focusing on studies that have used TMS to monitor changes in the excitability of the corticospinal pathway. We discuss how these physiological results have motivated theoretical models of how the brain selects actions, regulates movement initiation and execution, and switches from one state to another.


[Incorporation of novel agents into the treatment for acute myeloid leukemia].
The mainstay of therapeutic modalities of acute myeloid leukemia (AML) includes intensive chemotherapies and allogeneic hematopoietic cell transplantation. The gold standard of the induction treatment is a regular dose of cytarabine plus anthracycline, and several courses of consolidation therapy are administered. Allogeneic hematopoietic cell transplantation is employed in patients with intermediate-poor risks. No new drugs have been introduced to the treatment of AML for nearly 30 years. However, in 2017, the US Food and Drug Administration approved four novel drugs for treating AML: FLT3 inhibitor midostaurin, IDH2 inhibitor enasidenib, liposomal cytarabine-daunorubicin CPX-351, and revived antibody-drug conjugate gemtuzumab ozogamicin. In Japan, several new agents are also undergoing clinical trials, including Bcl-2 inhibitor venetoclax, CDK9 inhibitor alvocidib, smoothened (SMO) inhibitor glasdegib, hypomethylating agents guadecitabine and azacitidine, NEDD8 inhibitor pevonedistat, and FLT3 inhibitors quizartinib and gilteritinib. These agents will be incorporated into the conventional 7+3 regimen or combined with hypomethylating agents or low-dose cytarabine to improve the therapeutic outcomes of AML.


Type II or type III radical hysterectomy compared to chemoradiotherapy as a primary intervention for stage IB2 cervical cancer.
Cervical cancer is the fourth most common cancer in women, with 528,000 estimated new cases globally in 2012. A large majority (around 85%) of the disease burden occurs in low- and middle-income countries (LMICs), where it accounts for almost 12% of all female cancers. Treatment of stage IB2 cervical cancers, which sit between early and advanced disease, is controversial. Some centres prefer to treat these cancers by radical hysterectomy, with chemoradiotherapy reserved for those at high risk of recurrence. In the UK, we treat stage IB2 cervical cancers mainly with chemoradiotherapy, based on the rationale that a high percentage will have risk factors necessitating chemoradiotherapy postsurgery. There has been no systematic review to determine the best possible evidence in managing these cancers. To determine if primary surgery for stage IB2 cervical cancer (type II or type III radical hysterectomy with lymphadenectomy) improves survival compared to primary chemoradiotherapy.To determine if primary surgery combined with postoperative adjuvant chemoradiotherapy, for stage IB2 cervical cancer increases patient morbidity in the management of stage IB2 cervical cancer compared to primary chemoradiotherapy. We searched the Cochrane Central Register of Controlled Trials (CENTRAL; 2018, Issue 3), MEDLINE via Ovid (1946 to April week 2, 2018) and Embase via Ovid (1980 to 2018 week 16). We also searched registers of clinical trials, abstracts of scientific meetings and reference lists of included studies up to April 2018. We searched for randomised controlled trials (RCTs), quasi-RCTs or non-randomised studies (NRSs) comparing surgery to chemoradiotherapy in stage IB2 cervical cancers. Two review authors independently assessed whether potentially relevant studies met the inclusion criteria, abstracted data, assessed risk of bias and analysed data using standard methodological procedures expected by Cochrane. We identified 4968 records from the literature searches, but we did not identify any RCTs that compared primary surgery with chemoradiotherapy in stage IB2 cervical cancer.We found one NRS comparing surgery to chemoradiotherapy in IB2 and IIA2 cervical cancers which met the inclusion criteria. However, we were unable to obtain data for stage IB2 cancers only and considered the findings very uncertain due to a high risk of selection bias. There is an absence of high-certainty evidence on the relative benefits and harms of primary radical hysterectomy versus primary chemoradiotherapy for stage IB2 cervical cancer. More research is needed on the different treatment options in stage IB2 cervical cancer, particularly with respect to survival, adverse effects, and quality of life to facilitate informed decision-making and individualised care.


Comparison of intraoperative basal fluid requirements in distal pancreatectomy: Laparotomy vs. laparoscopy: A retrospective cohort study.
There has been recent progress in intraoperative fluid therapy. However, little is known about intraoperative fluid therapy in laparoscopic surgery. The purpose of this study is to determine whether there are differences in the basal fluid requirements during surgery between laparotomy and laparoscopic distal pancreatectomy.This retrospective cohort study analyzed the electronic medical records of 253 patients who underwent distal pancreatectomy via either laparotomy (73 patients) or laparoscopy (180 patients) between June 2006 and March 2016. The volume of intraoperative fluid administered, postoperative complications, length of hospital stay, and readmission rate were evaluated. The total volume of fluids was calculated as the sum of the volume of crystalloid plus the volume of colloid multiplied by 1.5 or 2.0.Patients who had laparotomy were older and had higher American Society of Anesthesiologists classes. Anesthesia time was longer and estimated blood loss was larger in laparotomy. More colloid (1.8 mL/kg per h vs. 1.2 mL/kg per h, P < .001) and more total calculated fluid (1.5 times: 11.7 mL/kg per h vs. 10.6 mL/kg per h, P = .002; 2.0 times: 12.6 mL/kg per h vs. 11.2 mL/kg per h, P = .001) were infused in laparotomy. Crystalloid (9.0 mL/kg per h vs. 8.9 mL/kg per h, P = .203) did not show significant difference. Postoperative complications were more frequent (63% vs. 45%, P = .008), the hospital stay was longer (18 days vs. 13.4 days, P < .001), and readmission rate was higher (15% vs. 5.6%, P = .02) in laparotomy. By logistic regression analysis, we could find that operation type (laparotomy vs. laparoscopy, odds ratio 1.900, 95% confidence interval 1.072-3.368) and operation time (P = .004) had effect on complications.In patients undergoing distal pancreatectomy, basal fluid requirements were larger in laparotomy compared with laparoscopy. Operation time and estimated blood loss had effects on fluid administration. Postoperative complications were more frequent in laparotomy but we could not find relationships with infused colloid or total calculated fluid volumes. Operation type (laparotomy vs. laparoscopy) and operation time were the only related factors to postoperative complications.


Drug Regulatory-Compliant Validation of a qPCR Assay for Bioanalysis Studies of a Cell Therapy Product with a Special Focus on Matrix Interferences in a Wide Range of Organ Tissues.
Quantitative polymerase chain reaction (qPCR) has emerged as an important bioanalytical method for assessing the pharmacokinetics of human-cell-based medicinal products after xenotransplantation into immunodeficient mice. A particular challenge in bioanalytical qPCR studies is that the different tissues of the host organism can affect amplification efficiency and amplicon detection to varying degrees, and ignoring these matrix effects can easily cause a significant underestimation of the true number of target cells in a sample. Here, we describe the development and drug regulatory-compliant validation of a TaqMan® qPCR assay for the quantification of mesenchymal stromal cells in the range of 125 to 20,000 cells/200 µL lysate via the amplification of a human-specific, highly repetitive α-satellite DNA sequence of the chromosome 17 centromere region HSSATA17. An assessment of matrix effects in 14 different mouse tissues and blood revealed a wide range of spike recovery rates across the different tissue types, from 11 to 174%. Based on these observations, we propose performing systematic spike-and-recovery experiments during assay validation and correcting for the effects of the different tissue matrices on cell quantification in subsequent bioanalytical studies by multiplying the back-calculated cell number by tissue-specific factors derived from the inverse of the validated percent recovery rate.


Cisplatin Binding Sites in Human H-Chain Ferritin.
The aim of this work is to identify the cisplatin binding sites on human H-chain ferritin. High-resolution X-ray crystallography reveals that cisplatin binds four distinct protein sites, that is, the side chains of His136 and Lys68, the side chain of His105, the side chain of Cys90 and the side chain of Cys102. These Pt binding sites are compared with those observed for the adduct that cisplatin forms upon encapsulation within horse spleen L-chain ferritin (87% identity with human L-chain ferritin).


Reduction-sensitive CD44 receptor-targeted hyaluronic acid derivative micelles for doxorubicin delivery.
A reduction-sensitive CD44-positive tumor-targetable drug delivery system for doxorubicin (DOX) delivery was developed based on hyaluronic acid (HA)-grafted polymers. HA was conjugated with folic acid (FA) via a reduction-sensitive disulfide linkage to form an amphiphilic polymer (HA-ss-FA). The chemical structure of HA-ss-FA was analyzed by ultraviolet spectroscopy, Fourier transform infrared spectroscopy, and 1H nuclear magnetic resonance (NMR) spectroscopy. The molecular weight of HA-ss-FA was determined by high-performance gel permeation chromatography. Blank HA-ss-FA micelles and DOX-loaded micelles were prepared and characterized. The reduction responsibility, cellular uptake, and in vivo biodistribution of HA-ss-FA micelles were investigated. DOX-loaded micelles were of high encapsulation efficiency (88.09%), high drug-loading content (22.70%), appropriate mean diameter (100-120 nm), narrow size distribution, and negative zeta potential (-6.7 to -31.5 mV). The DOX release from the micelles was significantly enhanced in reduction environment compared to normal environment. The result of in vitro cytotoxicity assay indicated that the blank micelles were of low toxicity and good biocompatibility and the cell viabilities were >100% with the concentration of HA-ss-FA from 18.75 to 600.00 μg/mL. Cellular uptake and in vivo biodistribution studies showed that DOX-loaded micelles were tumor-targetable and could significantly enhance cellular uptake by CD44 receptor-mediated endocytosis, and the cellular uptake of DOX in CD44-positve A549 cells was 1.6-fold more than that in CD44-negative L02 cells. In vivo biodistribution of HA-ss-FA micelles showed that micelles were of good in vivo tumor targetability and the fluorescence of indocyanine green (ICG)-loaded micelles was 4- to 6.6-fold stronger than free ICG within 6 h in HCCLM3 tumor-bearing nude mice. HA-ss-FA is a promising nanocarrier with excellent biocompatibility, tumor targetability, and controlled drug release capability for delivery of chemotherapy drugs in cancer therapy.


Benzene Exposure From Selected Work Tasks on Offshore Petroleum Installations on the Norwegian Continental Shelf, 2002-2018.
Work on offshore petroleum installations may cause exposure to benzene. Benzene is a carcinogenic agent, and exposure among workers should be as low as reasonably practicable. We aimed to assess short-term (less than 60 min) benzene exposure from the most frequent work tasks on offshore installations on the Norwegian continental shelf and identify determinants of exposure. In addition, we aimed to assess the time trend in task-based benzene measurements from 2002 to 2018. The study included 763 task-based measurements with a sampling duration of less than 60 min, collected on 28 offshore installations from 2002 to 2018. The measurements were categorized into 10 different tasks. Multilevel mixed-effect Tobit regression models were developed for two tasks: sampling and disassembling/assembling equipment. Benzene source, season, indoors or outdoors, design of process area, year of production start, sampling method, and work operation were considered as potential determinants for benzene exposure in the models. The overall geometric mean (GM) benzene exposure was 0.02 ppm (95% confidence intervals 95%(CI: 0.01-0.04). The pipeline inspection gauge (PIG) operation task was associated with the highest exposure, with a GM of 0.33 ppm, followed by work on flotation cells, disassembling/assembling, and sampling, with GMs of 0.16, 0.04, and 0.01 ppm, respectively. Significant determinants for the disassembling/assembling task were work operation (changing or recertifying valves, changing or cleaning filters, and breaking pipes) and benzene source. For sampling, the benzene source was a significant determinant. Overall, the task-based benzene exposure declined annually by 10.2% (CI 95%: -17.4 to -2.4%) from 2002 to 2018. The PIG operation task was associated with the highest exposure out of the ten tasks, followed by work on flotation cells and when performing disassembling/assembling of equipment. The exposure was associated with the type of benzene source that was worked on. Despite the decline in task-based exposure in 2002-2018, technical measures should still be considered in order to reduce the exposure.


Combination treatment of dupilumab with bortezomib in a patient with IgG kappa gammopathy of renal significance, uremic pruritus and chronic lichenoid dermatitis.
Chronic pruritus (CP) is one of the most frequent symptoms among dermatological conditions, capable of reducing the quality of life (QoL). CP may be induced by atopic dermatitis or other dermatological and/or non-dermatological conditions. In this article, we report the case of a patient affected by generalized CP, characterised by multiple papulo-nodular and escoriatic lesions, developed after the onset of an immunoglobulin G (IgG) kappa monoclonal gammopathy of renal significance (MGRS), associated with renal insufficiency. Therefore, a combined treatment with dupilumab for CP and bortezomib for the hematologic malignancy was administered to the patient. The present case report highlights the efficacy of dupilumab for the treatment of CP. Moreover, no relevant side effects were recorded during the treatment in combination with other systemic biological drugs for other systemic pathologies.


Xanthobacter oligotrophicus sp.nov., isolated from paper mill sewage.
A novel, aerobic nitrogen-fixing methylotrophic bacterium, strain 29kT, was enriched and isolated from sludge generated during wastewater treatment at a paper mill in Baikal, Russian Federation. Cells were Gram-stain-variable. The cell wall was of the negative Gram-type. Cells were curved oval rod-shaped, 0.5-0.7×1.7-3.4 µm and formed yellow-coloured colonies. Cells tended to be pleomorphic if grown on media containing succinate or coccoid if grown in the presence of methyl alcohol as the sole carbon source. Cells were non-motile, non-spore-forming and contained retractile (polyphosphate) and lipid (poly-β-hydroxybutyrate) bodies. The major respiratory quinone was ubiquinone Q-10 and the predominant cellular fatty acids were C18:1 ω7, C19:0 cyclo and C16:0. The genomic DNA G+C content was 67.95 mol%. Strain 29kT was able to grow at 4-37 °C (optimum, 30 °C), at pH 6.0-8.5 (optimum, pH 6.5-7.0) and at salinities of 0-0.5% (w/v) NaCl (optimum, 0% NaCl). Catalase and oxidase were positive. Strain 29kT could grow chemolithoautotrophically in mineral media under an atmosphere of H2, O2 and CO2 as well as chemoorganoheterotrophically on methanol, ethanol, n-propanol, n-butanol and various organic acids. The carbohydrate utilization spectrum is limited by glucose and raffinose. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the newly isolated strain was a member of the genus Xanthobacter with Xanthobacter autotrophicus 7cT (99.9% similarity) and Xanthobacter viscosus 7dT (99.4 % similarity) as closest relatives among species with validly published names. The average nucleotide identity and digital DNA-DNA hybridization values of 92.7 and 44.9%, respectively, of the 29kT to the genome of the most closely related species, X. autotrophicus 7cT, were below the species cutoffs. Based on genotypic, phenotypic and chemotaxonomic characteristics, it is proposed that the isolate represents a novel species, Xanthobacter oligotrophicus sp. nov. The type strain is 29kT (=KCTC 72777T=VKM B-3453T).


Phase II, randomized, double-blind, placebo-controlled, multicenter study to investigate the immunogenicity and safety of a West Nile virus vaccine in healthy adults.
ChimeriVax-WN02 is a live, attenuated chimeric vaccine for protection against West Nile virus. This Phase II, randomized, double-blind, placebo-controlled, multicenter study assessed the immunogenicity, viremia, and safety of the ChimeriVax-WN02 vaccine. The 2-part study included adults in general good health. In part 1, subjects aged 18-40 years were randomized to 1 of 4 treatment groups: ChimeriVax-WN02 3.7- × -10(5) plaque-forming units (PFU), 3.7 × 10(4) PFU, 3.7 × 10(3) PFU, or placebo. In part 2, subjects aged 41-64 and ≥ 65 years were randomized to receive ChimeriVax-WN02 3.7 × 10(5) PFU or placebo. In both part 1 and part 2, seroconversion was achieved at day 28 by >96% of subjects in active treatment groups. In part 1, neutralizing antibody titers at day 28 were higher and viremia levels lower with the highest dose, whereas the adverse event profile was similar between the dose groups. In part 2, antibody titers and viremia levels were higher in subjects aged ≥ 65 years, and more subjects in the 41-64 years cohort experienced adverse events. The ChimeriVax-WN02 vaccine was highly immunogenic in younger adults and the elderly, and it was well tolerated at all dose levels and in all age groups investigated. Clinical Trials.gov identifier: NCT00442169.


Heat exchange following atropine injection before and after heat acclimation.
The effect of saline and atropine injection (2 mg, im) on eight healthy male subjects before and after heat acclimation was studied while each subject walked on a treadmill (1.34 m X s-1) in a hot-dry environment (ambient temperature = 48.4 degrees C, dew-point temperature = 20.5 degrees C). Partitional calorimetric analysis was done for the periods in which maximum sweat inhibition occurred (30 min). Mean skin temperature, rectal temperature, and heart rate were continuously observed. Evaporative loss from the skin was calculated by changes in body weight (Sauter balance); heat transfer coefficients were defined by Nishi equations. A prediction of sweat inhibition based on an analysis of heat storage and its effect on a theoretical temperature, which can be graphed on a psychrometric chart, was developed. A rational effective temperature (ET) defined as the operative temperature at the intersection of the 50% rh, which encompasses total heat exchange, was used to compare the effects of atropine before and after heat acclimation. The results show that heat acclimation reduced ET by approximately 2.5 degrees C when compared with the unacclimated state after atropine injection. Thus heat acclimation reduces the hazards of heatstroke caused by exercise in the heat with atropine injection.


Altered lymph node composition in diphtheria toxin receptor-based mouse models to ablate dendritic cells.
Dendritic cells (DCs) are key regulators of innate and adaptive immunity. Our understanding of immune function has benefited greatly from mouse models allowing for selective ablation of DCs. Many such models rely on transgenic diphtheria toxin receptor (DTR) expression driven by DC-restricted promoters. This renders DCs sensitive to DT but is otherwise thought to have no effect on immune physiology. In this study, we report that, unexpectedly, mice in which DTR is expressed on conventional DCs display marked lymph node (LN) hypocellularity and reduced frequency of DCs in the same organs but not in spleen or nonlymphoid tissues. Intriguingly, in mixed bone marrow chimeras the phenotype conferred by DTR-expressing DCs is dominant over control bone marrow-derived cells, leading to small LNs and an overall paucity of DCs independently of the genetic ability to express DTR. The finding of alterations in LN composition and size independently of DT challenge suggests that caution must be exercised when interpreting results of experiments obtained with mouse models to inducibly deplete DCs. It further indicates that DTR, a member of the epidermal growth factor family, is biologically active in mice. Its use in cell ablation experiments needs to be considered in light of this activity.


Primary osteogenic sarcoma of the breast.
Primary osteogenic sarcomas of the breast are exceptionally uncommon. We describe such a case occurring in a 66-year-old woman who presented with a hard mass in her left breast. Mammography and breast ultrasonography showed a calcified breast lump, but features were not diagnostic. Modified radical mastectomy of the left breast, including axillary lymph node dissection, was performed. Microscopical and immunohistochemical findings established the diagnosis of primary osteogenic sarcoma. Because there was no evidence of metastasis, no further treatment was considered necessary. She remained well 15 months later, without tumour recurrence. We discuss in detail the diagnostic implications of this rare entity.


Progressive hypokalaemia in elderly patients taking three thiazide potassium-sparing diuretic combinations for thirty-six months.
Three different thiazide potassium-sparing diuretic combinations were given to elderly patients for heart failure. Eighty patients received their allocated combinations for 3 years and had 6-monthly measurements of plasma potassium. A further 84 were recruited for study but 29 died within 6 months and 55 had to be withdrawn from the trial. The triamterene-containing preparation was discontinued most frequently (6/44) because of hypokalaemia (plasma potassium less than 3.0 mmol/l); amiloride (5/44) and spironolactone (1/47). The median fall in plasma potassium over 3 years in those patients not withdrawn because of hypokalaemia was similar in each case (P greater than 0.05) and possibly failed to reach significance because of the withdrawal rate (9%). The trend was for a greater fall in those patients taking triamterene. The spironolactone-containing preparation may be the least unsatisfactory of the three.


[Sonographic semiotics of ureterohydronephrosis in children].
To identify typical sonographic semiotics of ureterohydronephrosis in children having importance in determining the severity of disease and the choice of treatment strategy in this category of patients. Sonographic images of the kidneys and urinary tract of 158 children with ureterohydronephrosis were examined. Unilateral and bilateral ureterohydronephrosis was found in 75 and 83 children, respectively. There were 100 (63.3%) boys and 58 (36.7%) girls. The age of patients ranged from 3 months to 14 years (mean 4.15+/-3.21). Ultrasound examination enables the upper 193 (80.1%) and lower 167 (69.3%) third of ureters to be visualized. At the same time more than half of the visualized ureters had a diameter greater than 1 cm, and one-tenth of the affected ureters had diameters greater than 2 cm. If direct visualization of the ureters was impossible, pelvicalyceal system was evaluated, which was expanded on the affected side in all examined children. The morpho-sonographic changes of renal parenchymal in children with III-IV degree ureterohydronephrosis are described in details. Ultrasound examination can be regarded as a screening method for initial evaluation of patients with ureterohydronephrosis. It provides a rough estimation of the extent, level and nature of disturbances of urinary excretion, a preliminary assessment of the renal parenchyma and identification of anomalies of the urinary tract. These findings can be useful in choosing an optimal algorithm of diagnosis and surgical treatment of the disease.


The evolution of chromosomal sex determination.
There is a great diversity of sex determination mechanisms, with evidence for numerous evolutionary transitions between different systems. For example, environmental sex determination is widespread in lower vertebrates, and genetic sex determination has probably evolved from it several times. This requires the establishment of genes that override environmental cues. Close linkage between male and female determining loci is favoured by selection, and represents the first step towards the evolution of highly differentiated sex chromosomes. Once crossing over between primitive sex chromosomes has been suppressed, the primitive Y (W) chromosome is vulnerable to the operation of forces that lead to a reduction in its effective population size. This reduces the ability of natural selection to maintain the functionality of genes on the proto-Y, so that it gradually degenerates. Primitive sex chromosome systems, and systems of neo-X and neo-Y chromosomes formed by translocations involving autosomes and sex chromosomes, provide an opportunity to test evolutionary models of the degeneration of Y chromosomes and to determine the time-scales involved. Recent data confirm that newly-evolving Y or neo-Y chromosomes experience a sharp reduction in effective population size, and indicate that degeneration can occur over a few million generations.


[Calcimimetic drugs in nondialyzed patients].
With the progression of chronic kidney disease (CKD) possibilities of regulation of parathyroid hormone (PTH) secretion also decrease. Calcimimetics offer the possibility of direct influence on pathogenic mechanisms of secondary hyperparathyroidism in the early stages of CKD. Type II calcimimetic drugs inhibit PTH secretion by sensitizing the parathyroid calcium receptor to extracellular calcium. They found clinical application of in disease states running from hyperparathyroidism. The paper discusses the application of calcimimetic drug type II--cinacalcet--in the treatment of secondary hyperparathyroidism in patients with CKD and patients with hyperparathyroidism surviving renal transplantation. Cinacalcet oral administration (15-180 mg/day) significantly reduces plasma PTH concentration (> or = 30% as compared to baseline values) at a relatively low side effects (hypocalcemia, hyperphosphatemia, gastrointestinal symptoms). In patients after renal transplantation cinacalcet, according to the majority of data, has no effect on graft function and effectively helps in the treatment of persistent hyperparathyroidism, especially running with hypercalcemia.