PeBL1, a novel protein elicitor from Brevibacillus laterosporus strain A60, activates defense responses and systemic resistance in Nicotiana benthamiana.
We report the identification, characterization, and gene cloning of a novel protein elicitor (PeBL1) secreted from Brevibacillus laterosporus strain A60. Through a purification process consisting of ion-exchange chromatography and high-performance liquid chromatography (HPLC), we isolated a protein that was identified by electrospray ionization quadrupole time of flight tandem mass spectrometry (ESI-Q-TOF-MS-MS). The 351-bp PeBL1 gene produces a 12,833-Da protein with 116 amino acids that contains a 30-residue signal peptide. The PeBL1 protein was expressed in Escherichia coli. The recombinant protein can induce a typical hypersensitive response (HR) and systemic resistance in Nicotiana benthamiana, like the endogenous protein. PeBL1-treated N. benthamiana exhibited strong resistance to the infection of tobacco mosaic virus-green fluorescent protein (TMV-GFP) and Pseudomonas syringae pv. tabaci compared to control N. benthamiana. In addition, PeBL1 triggered a cascade of events that resulted in defense responses in plants, including reactive oxygen species (ROS) production, extracellular-medium alkalization, phenolic-compound deposition, and expression of several defense-related genes. Real-time quantitative-PCR analysis indicated that the known defense-related genes PR-1, PR-5, PDF1.2, NPR1, and PAL were upregulated to varying degrees by PeBL1. This research not only provides insights into the mechanism by which beneficial bacteria activate plant systemic resistance, but also sheds new light on a novel strategy for biocontrol using strain A60.


Periprocedural management of patients receiving novel oral anticoagulants.
Non-vitamin K oral antagonists are being increasingly used. However, broad clinical experience with them is lacking. To review guidelines and evidence for the use of non-vitamin K oral antagonists in the periprocedural environment. Despite the clear advantages of vitamin K oral antagonists, their use can entail risks owing to the scarcity of reversal agents. Consensus has been reached about postoperative resumption, which is recommended at 24 hours and 48-72 hours, respectively, after low-risk and high-risk bleeding surgery. Bridging with heparin is recommended in patients with a high risk of thrombosis. Urgent interventions should ideally take place 24 hours after the last dose intake. Major discrepancies exist between the American and the European recommendations for neuraxial procedures. The American proposals recommend suspending the drug for five half-lives, whereas the European approaches suggest suspension of just two half-lives. Suggestions for perioperative discontinuation vary widely. Some authors recommend a longer time of resumption for patients with renal impairment. All agree that there should be an increase in the number of days of interruption in high-risk bleeding procedures versus low-risk bleeding procedures. A diverse number of approaches have been suggested for perioperative management of novel oral antagonists. American recommendations tend to be more rigorous than those of Europe. A need for more studies that measure health outcomes after the use of these drugs would be indispensable.


Anaerobic carbon monoxide dehydrogenase diversity in the homoacetogenic hindgut microbial communities of lower termites and the wood roach.
Anaerobic carbon monoxide dehydrogenase (CODH) is a key enzyme in the Wood-Ljungdahl (acetyl-CoA) pathway for acetogenesis performed by homoacetogenic bacteria. Acetate generated by gut bacteria via the acetyl-CoA pathway provides considerable nutrition to wood-feeding dictyopteran insects making CODH important to the obligate mutualism occurring between termites and their hindgut microbiota. To investigate CODH diversity in insect gut communities, we developed the first degenerate primers designed to amplify cooS genes, which encode the catalytic (β) subunit of anaerobic CODH enzyme complexes. These primers target over 68 million combinations of potential forward and reverse cooS primer-binding sequences. We used the primers to identify cooS genes in bacterial isolates from the hindgut of a phylogenetically lower termite and to sample cooS diversity present in a variety of insect hindgut microbial communities including those of three phylogenetically-lower termites, Zootermopsis nevadensis, Reticulitermes hesperus, and Incisitermes minor, a wood-feeding cockroach, Cryptocercus punctulatus, and an omnivorous cockroach, Periplaneta americana. In total, we sequenced and analyzed 151 different cooS genes. These genes encode proteins that group within one of three highly divergent CODH phylogenetic clades. Each insect gut community contained CODH variants from all three of these clades. The patterns of CODH diversity in these communities likely reflect differences in enzyme or physiological function, and suggest that a diversity of microbial species participate in homoacetogenesis in these communities.


[Oral vitamin A tolerance tests in hypertriglyceridemia (author's transl)].
HVA tolerance tests were performed to study the post-absorptive lipemia of 32 patients with hypertriglyceridemia. Among them 10 had a normal test and 22 a pathological test. This difference shows the existence of an exogenous factor in the second group of hypertriglyceridemias, which is poorly related to the typing of the WHO classification, but much more to the value of the cholesterol/triglycerides ratio of each patient. The separation of these two groups is of great interest for their therapeutical management, the hypertriglyceridemias with exogenous factor being more sensitive to diet than the others.


Sorting of sphingolipids in the endocytic pathway of HT29 cells.
The intracellular flow and fate of two fluorescently labeled sphingolipids, 6-[N-(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]hexanoyl glucosyl sphingosine (C6-NBD-glucosylceramide) and C6-NBD-sphingomyelin, was examined in the human colon adenocarcinoma cell line HT29. After their insertion into the plasma membrane at low temperature and subsequent warming of the cells to 37 degrees C, both sphingolipid analogues were internalized by endocytosis, but their intracellular site of destination differed. After 30 min of internalization, C6-NBD-glucosylceramide was localized in the Golgi apparatus, as demonstrated by colocalization with fluorescently labeled ceramide, a Golgi complex marker, and by showing that monensin-induced disruption of the Golgi structure was paralleled by a similar perturbation of the fluorescence distribution. By contrast, C6-NBD-sphingomyelin does not colocalize with the tagged ceramide. Rather, a colocalization with ricin, which is internalized by endocytosis and predominantly reaches the lysosomes, was observed, indicating that the site of delivery of this lipid is restricted to endosomal/lysosomal compartments. Also, in monensin-treated cells no change in the distribution of fluorescence was observed. Thus, these results demonstrate that (sphingo)lipid sorting can occur in the endocytic pathway. Interestingly, the observed sorting phenomenon was specific for glucosylceramide, when compared to other glycolipids, while only undifferentiated HT29 cells displayed the different routing of the two lipids. In differentiated HT29 cells the internalization pathway of sphingomyelin and glucosylceramide was indistinguishable from that of transferrin.


Diversity of microbial communities in an attached-growth system using Bauxsol™ pellets for wastewater treatment.
Columns of Bauxsol™ pellets were used in a field experiment as biomass support particle for wastewater microbial communities. The attached microbial community structure was analysed using denaturing gradient gel electrophoresis (DGGE), targeting the 16S rDNA gene's V3 region. DGGE profiles showed that the type and composition of support particles used (i.e. Bauxsol™ pellets or gravel) had a significant impact on the attached bacterial communities (64% dissimilarity). In addition, ecological indices revealed a more heterogeneous bacterial community structure on the Bauxsol™ pellets. TOC/TN ratios post-experiment (6.5-9.3) suggested a good level of biological activity (i.e. active biofilm) in the Bauxsol™ columns. Moreover, Bauxsol™ pellets were mostly made of inorganic carbon, suggesting insoluble carbonate biomineralisation. Polymerase chain reaction (PCR) amplification of specific marker genes (i.e. bacterial and archaeal amoA genes, nosZ gene, and hzo gene) were used to identify the presence of attached bacterial communities associated with nitrogen transformation. The results along with geochemical data (i.e. up to 50% nitrogen removal) revealed co-existence of ammonia-oxidising bacteria, denitrifiers, and anammox organisms. This study conclusively demonstrates that microbial communities are well-adapted to Bauxsol™ pellets and bacterial communities involved in the nitrogen cycle are present.


Complex interplay between the lipin 1 and the hepatocyte nuclear factor 4 α (HNF4α) pathways to regulate liver lipid metabolism.
Lipin 1 is a bifunctional protein that serves as a metabolic enzyme in the triglyceride synthesis pathway and regulates gene expression through direct protein-protein interactions with DNA-bound transcription factors in liver. Herein, we demonstrate that lipin 1 is a target gene of the hepatocyte nuclear factor 4α (HNF4α), which induces lipin 1 gene expression in cooperation with peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) through a nuclear receptor response element in the first intron of the lipin 1 gene. The results of a series of gain-of-function and loss-of-function studies demonstrate that lipin 1 coactivates HNF4α to activate the expression of a variety of genes encoding enzymes involved in fatty acid catabolism. In contrast, lipin 1 reduces the ability of HNF4α to induce the expression of genes encoding apoproteins A4 and C3. Although the ability of lipin to diminish HNF4α activity on these promoters required a direct physical interaction between the two proteins, lipin 1 did not occupy the promoters of the repressed genes and enhances the intrinsic activity of HNF4α in a promoter-independent context. Thus, the induction of lipin 1 by HNF4α may serve as a mechanism to affect promoter selection to direct HNF4α to promoters of genes encoding fatty acid oxidation enzymes.


Multi-scale dynamic imaging reveals that cooperative motility behaviors promote efficient predation in bacteria.
Many species, such as fish schools or bird flocks, rely on collective motion to forage, prey, or escape predators. Likewise, Myxococcus xanthus forages and moves collectively to prey and feed on other bacterial species. These activities require two distinct motility machines enabling adventurous (A) and social (S) gliding, however when and how these mechanisms are used has remained elusive. Here, we address this long-standing question by applying multiscale semantic cell tracking during predation. We show that: (1) foragers and swarms can comprise A- and S-motile cells, with single cells exchanging frequently between these groups; (2) A-motility is critical to ensure the directional movement of both foragers and swarms; (3) the combined action of A- and S-motile cells within swarms leads to increased predation efficiencies. These results challenge the notion that A- and S-motilities are exclusive to foragers and swarms, and show that these machines act synergistically to enhance predation efficiency.


A physiological trait-based approach to predicting the responses of species to experimental climate warming.
Physiological tolerance of environmental conditions can influence species-level responses to climate change. Here, we used species-specific thermal tolerances to predict the community responses of ant species to experimental forest-floor warming at the northern and southern boundaries of temperate hardwood forests in eastern North America. We then compared the predictive ability of thermal tolerance vs. correlative species distribution models (SDMs) which are popular forecasting tools for modeling the effects of climate change. Thermal tolerances predicted the responses of 19 ant species to experimental climate warming at the southern site, where environmental conditions are relatively close to the ants' upper thermal limits. In contrast, thermal tolerances did not predict the responses of the six species in the northern site, where environmental conditions are relatively far from the ants' upper thermal limits. Correlative SDMs were not predictive at either site. Our results suggest that, in environments close to a species' physiological limits, physiological trait-based measurements can successfully forecast the responses of species to future conditions. Although correlative SDMs may predict large-scale responses, such models may not be accurate for predicting site-level responses.


Desire thinking as a mediator of the relationship between novelty seeking and craving.
The construct of craving has been shown to play a crucial role in the development and maintenance of addictive behaviors. Both novelty seeking and desire thinking have been identified, respectively, as important temperamental and cognitive predictors of craving. In the present study we aimed to explore the relative contribution of novelty seeking and desire thinking towards craving, hypothesizing a sequence of multiple mediating relationships starting from novelty seeking and moving onto imaginal prefiguration, verbal perseveration and craving in serial fashion. A convenience sample of 270 individuals completed measures assessing novelty seeking, desire thinking, and craving relating to a chosen activity. Findings showed that, controlling for age and gender, desire thinking components predicted craving over and above novelty seeking. The indirect effect from novelty seeking to craving, via desire thinking components, was significant thus supporting a multiple-mediational sequence. Finally, the relationship between imaginal prefiguration and craving was found to be partially mediated by verbal perseveration. The findings provide support for the conceptualization of desire thinking as an independent construct in predicting craving over and above novelty seeking.


Scalable synthesis of SnS2/S-doped graphene composites for superior Li/Na-ion batteries.
Tin disulfide (SnS2) has emerged as a promising anode material for lithium/sodium ion batteries (LIBs/SIBs) due to its unique layered structure, outstanding electrochemical properties and low cost. However, its poor cycling life and time-consuming synthesis as well as low-yield production hinder the practical utilization of nanostructured SnS2. In this work, we demonstrate a simple and reliable dissolution-regeneration strategy to construct a flexible SnS2/sulfur-doped reduced graphene oxide (S-rGO) composite as anodes for LIBs and SIBs, highlighting its mass-production feature. In addition, the robust affinity between SnS2 and S-rGO without interstitial volume is very beneficial for preventing the SnS2 particles from breaking themselves away from the rGO nanosheets into free nanoparticles. As a result, the SnS2/S-rGO composite as anodes delivers high reversible capacities of 1078 mA h g-1 and 564 mA h g-1 (at 0.1 A g-1) for LIBs and SIBs, respectively, and excellent rate capabilities and cycling stability (e.g. 532 mA h g-1 during the 600 cycles at 5.0 A g-1 for LIBs). Our proposed strategy may also possess great potential for the practical application of other electrochemically active metal sulfide composites for energy devices.


Dosimetric Feasibility Study of Dose Escalated Stereotactic Body Radiation Therapy (SBRT) in Locally Advanced Pancreatic Cancer (LAPC) Patients: It Is Time to Raise the Bar.
To assess the dosimetric feasibility of a stereotactic body radiotherapy (SBRT) dose escalated protocol, with a simultaneous integrated boost (SIB) and a simultaneous integrated protection (SIP) approach, in patients with locally advanced pancreatic cancer (LAPC). Twenty LAPC lesions, previously treated with SBRT at our Institution, were re-planned. The original prescribed and administered dose was 50/30/25 Gy in five fractions to PTVsib (tumor-vessel interface [TVI])/PTVt (tumor volume)/PTVsip (overlap area between PTVt and planning organs at risk volume [PRVoars]), respectively. At re-planning, the prescribed dose was escalated up to 60/40/33 Gy in five fractions to PTVsib/PTVt/PTVsip, respectively. All plans were performed using an inspiration breath hold (IBH) technique and generated with volumetric modulated arc therapy (VMAT). Well-established and accepted OAR dose constraints were used (D0.5cc < 33 Gy for luminal OARs and D0.5cc < 38 Gy for corresponding PRVoars). The primary end-point was to achieve a median dose equal to the prescription dose for the PTVsib with D98≥ 95% (95% of prescription dose is the minimum dose), and a coverage for PTVt and PTVsip of D95≥95%, with minor deviations in OAR dose constraints in < 10% of the plans. PTVsib median (± SD) dose/D95/conformity index (CI) were 60.54 (± 0.85) Gy/58.96 (± 0.86) Gy/0.99 (± 0.01), respectively; whilst PTVt median (± SD) dose/D95 were 44.51 (± 2.69) Gy/38.44 (± 0.82) Gy, and PTVsip median (± SD) dose/D95 were 35.18 (± 1.42) Gy/33.01 (± 0.84) Gy, respectively. With regard to OARs, median (± SD) maximum dose (D0.5cc) to duodenum/stomach/bowel was 29.31 (± 5.72) Gy/25.29 (± 6.90) Gy/27.03 (± 5.67) Gy, respectively. A minor acceptable deviation was found for a single plan (bowel and duodenum D0.5cc=34.8 Gy). V38 < 0.5 cc was achieved for all PRV luminal OARs. In LAPC patients SBRT, with a SIB/SIP dose escalation approach up to 60/40/33 Gy in five fractions to PTVsib/PTVt/PTVsip, respectively, is dosimetrically feasible with adequate PTVs coverage and respect for OAR dose constraints.


[Certain aspects of the pathogenesis and prophylactic treatment of hypertensive crises].
The paper analyses the results of a study of the clinical peculiarities of the course of essential hypertension with crises, of the changes in the functional state of the central nervous system, and of some neuro-humoral systems of the human body. In most of the patients tending to develope frequent crises distinct changes were noted in the EEG that indicate dysfunction of the hypothalamic zone and of the reticular formation of the brain stem, their clinical course being characterized by significant astheno-neurotic disorders with autonomous dysfunction and cerebral angiodistonic disturbances. For the prevention of crises it is essential to conduct a pathogenetically substantiated and highly differentiated systematic therapy, as well as special measures aimed at increasing the endurance of the central nervous system, improving the circulation and metabolic processes in the brain, decreasing tnd hypothalamic structures of the brain, and correcting the dishormonal disorders.


The RNA-binding protein HuD regulates neuronal cell identity and maturation.
Neural Hu proteins (HuB/C/D) are RNA-binding proteins that have been shown to induce neuronal differentiation activity when overexpressed in immature neural progenitor cells or undifferentiated neuronal tumors. Newly generated HuD-deficient mice exhibited a transient impaired-cranial-nerve-development phenotype at an early embryonic stage. Adult HuD-deficient mice exhibited an abnormal hind-limb reflex and poor rotarod performance. Analysis of neurosphere formation revealed that the number and self-renewal capacity of the neural stem/progenitor cells were increased in HuD-deficient mice. HuD-deficient primary neurospheres also generated a smaller number of neurons. Cohort analysis of the cellular proliferative activity by using BrdUrd and iododeoxuridine labeling revealed that the number of differentiating quiescent cells in the embryonic cerebral wall was decreased. Long-term administration of BrdUrd revealed that the number of slowly dividing stem cells in the adult subventricular zone was increased in the HuD-deficient mice. Taken together, the results suggest that HuD is required at multiple points during neuronal development, including negative regulation of proliferative activity and neuronal cell-fate acquisition of neural stem/progenitor cells.


[Electromyographic features of 3 forms of myotonia].
On the basis of a great body of examination data the electromyographic peculiarities of three forms of myotonia are analyzed. As a result, the authors come to a conclusion that in the third form of myotonia the function of muscular fibres is grossly impaired, this impairment following the pattern of their being put out of activity. This failure appears invariably during the muscle activity and leads, in the end to the clinically observed transient, and in more advanced cases, moderately permanent weakness of the muscles and their hypotrophy. In addition, data on synaptic apparatus involvement were also obtained. The time course of the development of those disorders, as regards the neuromuscular conduction and the distal parts of the muscular fibres is followed. In atrophic myotonia, certain peculiarities of the electrophysiological characteristics of the neuromuscular periphery were noted. In particular, a marked phase of secondary refractoriness was revealed. Thomsen's myotonia was found to differ substantially in the electrophysiological characteristics from the above two forms of the myotonia. Although in a number of cases it is difficult to class a particular case with one or another form of myotonia (this can be explained by an exceedingly great diversity of the biochemical peculiarities inherited by each individual) the differentiation of the above three forms of myotonia is justified from the viewpoint of both exploring the pathogenesis of the disease forms, and choosing the treatment method the most effective in this particular case. Of importance in solving these problems may be diversified electrophysiological examinations of the motor apparatus, and comparisons of their results with those of clinical examinations.


Antioxidants and doxorubicin supplementation to modulate CD14 expression and oxidative stress induced by vitamin D3 and seocalcitol in HL60 cells.
1alpha,25-dihydroxyvitamin D3 (VD3) and the EB1089 analog are well known for their roles in the modulation of proliferation and the differentiation of several malignant cells. In addition, VD3 or EB1089 displayed a high disposal of oxidant features and the ability to cause release of reactive oxygen species (ROS). We attempted to enhance HL60 cell differentiation and to limit ROS generation, by the association of deltanoids with doxorubicin and the antioxidants catalase (CAT), superoxide dismutase (SOD) and N-acetyl cystein (NAC). Differentiation of HL60 cells into monocytic lineage was studied by expression of mRNA, protein CD14 and functional differentiation by the nitroblue tetrazolium assay. The 2',7'-dichlorodihydrofluorescein diacetate (H2-DCFDA) dye allowed to evaluate in situ ROS generation. When associated with 0.1 nM EB1089, 15 nM doxorubicin induced an increase of differentiated cell percentage from 29% to 87% and did not affect VD3-treated cells. The association with doxorubicin also induced a significant increase of ROS release (p<0.05) versus VD3 and EB1089-treated cells. These results correspond to additivity of individual effects of doxorubicin and deltanoids. Antioxidant agents (10 nM NAC, 50 U/ml SOD or 2000 U/ml CAT) were associated with 10 nM VD3 or 1 nM EB1089 for 72 h. Compared to VD3 and EB1089 treatments, associations with antioxidants induced a slight increase of differentiated cells and a significant increase of CD14 mRNA. The highest differentiation effect occurred in the case of the EB1089-NAC association. Antioxidants induced a decrease (p<0.05) in ROS release generated by VD3 or EB1089 near the level of untreated cells. Thus, antioxidant agents demonstrated a protective effect against VD3 and EB1089 oxidative cytotoxicity and an enhancement of the monocyte differentiation. Combinations of antioxidants with deltanoids could dissociate the oxidative stress and differentiation.


Citrulline level is a potent indicator of acute rejection in the long term following pediatric intestinal/multivisceral transplantation.
Citrulline has been advocated as a marker for acute cellular rejection (ACR) in intestinal transplantation; however, its significance as a forewarning in the long-term follow-up remains unknown. This study aimed to investigate the association between citrulline levels and the grading of ACR to establish a cutoff point that accurately predicts ACR beyond 3 months posttransplant in the pediatric patient population. During a 16-year period (1995-2011), a total of 13 499 citrulline samples were prospectively collected from 111 consecutive pediatric intestinal/multivisceral transplant recipients: 2155 were obtained concurrently with intestinal biopsies. There were 185 ACR episodes observed among 74/111 (67%) patients (median follow-up: 4.4 years). Citrulline levels were inversely proportional to the severity of ACR. Negative predictive values for any type of ACR (cutoff, 20 μmol/L) and moderate/severe ACR (cutoff, 10 μmol/L) were 95% and 99%, respectively. When patients were divided according to graft size, diagnostic accuracy using the same cutoff was identical. Similarly, subgroup analysis by the timing of citrulline measurement prior to biopsy varying from 1 to 7 days demonstrated comparable results. Citrulline is a potent indicator as a danger signal for ACR, being an exclusionary, noninvasive biomarker with excellent negative predictive values in the long term after pediatric intestinal/multivisceral transplant.


Depression, anxiety, and suicidal ideation in adults with COVID-induced parosmia.
KEY POINTS COVID-induced parosmia is associated with anxiety, depression, and suicidal ideation. Parosmic patients have low rates of treatment benefit and little hope for improvement. Hyposmia may mitigate the quality-of-life burden in patients with parosmia.


Analysis of α-synuclein species enriched from cerebral cortex of humans with sporadic dementia with Lewy bodies.
Since researchers identified α-synuclein as the principal component of Lewy bodies and Lewy neurites, studies have suggested that it plays a causative role in the pathogenesis of dementia with Lewy bodies and other 'synucleinopathies'. While α-synuclein dyshomeostasis likely contributes to the neurodegeneration associated with the synucleinopathies, few direct biochemical analyses of α-synuclein from diseased human brain tissue currently exist. In this study, we analysed sequential protein extracts from a substantial number of patients with neuropathological diagnoses of dementia with Lewy bodies and corresponding controls, detecting a shift of cytosolic and membrane-bound physiological α-synuclein to highly aggregated forms. We then fractionated aqueous extracts (cytosol) from cerebral cortex using non-denaturing methods to search for soluble, disease-associated high molecular weight species potentially associated with toxicity. We applied these fractions and corresponding insoluble fractions containing Lewy-type aggregates to several reporter assays to determine their bioactivity and cytotoxicity. Ultimately, high molecular weight cytosolic fractions enhances phospholipid membrane permeability, while insoluble, Lewy-associated fractions induced morphological changes in the neurites of human stem cell-derived neurons. While the concentrations of soluble, high molecular weight α-synuclein were only slightly elevated in brains of dementia with Lewy bodies patients compared to healthy, age-matched controls, these observations suggest that a small subset of soluble α-synuclein aggregates in the brain may drive early pathogenic effects, while Lewy body-associated α-synuclein can drive neurotoxicity.


Pharmacokinetics of orally administered uracil in healthy volunteers and in DPD-deficient patients, a possible tool for screening of DPD deficiency.
Dihydropyrimidine dehydrogenase (DPD) deficiency can lead to severe toxicity in patients treated with standard doses of 5-fluorouracil (5-FU). Oral uracil administration and subsequent measurement of uracil and dihydrouracil (DHU) plasma concentrations might detect patients with DPD deficiency. This study compares the pharmacokinetics of uracil and DHU after oral uracil administration in subjects with normal and deficient DPD status. Five hundred milligrams of uracil per metre square was administered orally to 11 subjects with normal DPD status and to 10 subjects with reduced DPD activity. Repeated administration (n = 3) of this dose was performed in 4 subjects, and 1,000 mg uracil/m(2) was administered to 4 subjects to assess intra-individual variation and linearity of pharmacokinetics. In subjects with normal DPD status, 500 mg/m(2) uracil resulted in uracil C (max) levels of 14.4 ± 4.7 mg/L at T (max) = 30.0 ± 11.6 min, and in DPD-deficient subjects, 20.0 ± 4.5 mg/L at 31.5 ± 1.1 min. The uracil AUC(0>180) was 31.2 ± 5.1 mg L/h in DPD-deficient subjects, which was significantly higher (P < 0.05) than in the subjects with normal DPD status (13.8 ± 3.9 mg L/h). Repeated uracil dosing showed reproducible uracil PK in subjects with normal DPD status, and dose elevation of uracil suggested linear pharmacokinetics. The pharmacokinetics of uracil differs significantly between subjects with a normal DPD activity and those with a deficient DPD status. The AUC and C (max) of uracil can be useful as a diagnostic tool to differentiate patients with regard to DPD status.


Integrated retrospective radiation dose assessment.
Radiation dose reconstruction is used to estimate exposure to radiation that has occurred externally, e.g., from an atomic bomb, or internally, e.g., from radionuclide ingestion. This commentary reviews techniques for biological dosimetry that have been developed to estimate radiation doses from internal exposures, but which can also be used to estimate external exposures. The author argues for increased development and use of these biological tools.


Searching of potential energy curves for the benzene dimer using dispersion-corrected density functional theory.
The present work aims to establish the utility of dispersion-corrected density functional theory for potential energy curves of the benzene dimer, a problem that has received significant attention for a long time. The interaction energies of parallel-stacked, T-shaped and parallel-displaced benzene dimer configurations have been evaluated using both dispersion- and normal gradient-corrected Perdew-Burke-Ernzerhof functionals along with Dunning's augmented correlation-consistent polarized valence triple-zeta (aug-cc-pVTZ) basis functions and compared with explicit correlation methods. The potential energy curves for the parallel-stacked and parallel-displaced benzene dimers are in excellent agreement with highly accurate coupled cluster (CCSD(T)) results, while for the T-shaped benzene dimer the dispersion-corrected results show a distinct deviation, being closer in that case to the MP2 level of results. The overestimation of interaction energy in the T-shaped dimer may be attributed to the presence of a permanent dipole moment in this configuration and indicates a structural dependence of the dispersion-corrected density functional method.


Evaluation of the potential of supplements to substitute for low-quality, tallgrass-prairie forage.
Thirteen ruminally fistulated steers (260 +/- 15 kg) were used in an incomplete Latin square with 13 treatments and four periods to evaluate the potential for different supplements to substitute for intake of low-quality, tallgrass-prairie forage. Steers were given ad libitum access to forage and received either no supplement (control = CTL) or one of four supplements, each fed at three different levels of intake. Supplements included 1) moderate CP (17.5%) concentrate (MCP-CON), 2) high CP (32.7%) concentrate (HCP-CON), 3) long-stem alfalfa hay (LSAH; 17.2% CP), and 4) alfalfa pellets (AP; 16.3% CP). Concentrates were mixtures of sorghum grain and soybean meal. Supplements were fed to supply .05, .10, and .15% BW of CP/d. Forage intake and digestible DMI were increased (P < .01) for supplemented steers compared with CTL (22 and 96%, respectively). Steers receiving increasing MCP-CON exhibited a quadratic (P = .03) forage intake response. Offering MCP-CON higher than .10% BW of CP/d (approximately .59% BW of DM) resulted in a substitution rate of -.56 g of forage/g of supplement. Although effects were not significant for steers receiving LSAH, the decline in forage intake at the high level of supplementation (-.48 g of forage/g of supplement) was similar in magnitude to that for MCP-CON. Forage intake increased linearly (P < .01) across supplementation levels for HCP-CON and AP supplements. Passage rates were faster (P < .01) for supplemented steers. However, passage rates for HCP-CON and AP groups increased linearly (P < .01) with increasing supplement, whereas they declined at the highest supplementation rate for MCP-CON and LSAH (quadratic, P < or = .05). Most fermentation variables displayed positive responses to supplementation per se and to increasing amount of supplements offered. In conclusion, although supplementation effectively enhances the use of low-quality forage, supplement type may affect the likelihood of observing substitution effects.


[Spontaneous and stimulated nitroblue tetrazolium reduction tests in tuberculosis of the lungs].
Cytochemical analysis of spontaneous and stimulated Nitro BT test values in 85 patients with pulmonary tuberculosis and in 24 ones with pneumonia has shown a significant increase of spontaneous Nitro BT test activity with the maximal values at the peak of the disease and an essential reduction of these values during remission. Stimulation with BCG vaccine results in a still more significant elevation of these parameters, depending on the depth of the involvement. Spontaneous and stimulated Nitro BT tests in patients with pulmonary tuberculosis and pneumonia appears to be a promising differential diagnostic test and a means for monitoring the therapy efficacy and for detection of secondary defects of phagocytosis system.


Ligand-induced IFNGR1 down-regulation calibrates myeloid cell IFNγ responsiveness.
The type II IFN (IFNγ) enhances antimicrobial activity yet also drives expression of genes that amplify inflammatory responses. Hence, excessive IFNγ stimulation can be pathogenic. Here, we describe a previously unappreciated mechanism whereby IFNγ itself dampens myeloid cell activation. Staining of monocytes from Listeria monocytogenes-infected mice provided evidence of type I IFN-independent reductions in IFNGR1. IFNγ was subsequently found to reduce surface IFNGR1 on cultured murine myeloid cells and human CD14+ peripheral blood mononuclear cells. IFNγ-driven reductions in IFNGR1 were not explained by ligand-induced receptor internalization. Rather, IFNγ reduced macrophage Ifngr1 transcription by altering chromatin structure at putative Ifngr1 enhancer sites. This is a distinct mechanism from that used by type I IFNs. Ligand-induced reductions in IFNGR1 altered myeloid cell sensitivity to IFNγ, blunting activation of STAT1 and 3. Our data, thus, reveal a mechanism by which IFNGR1 abundance and myeloid cell sensitivity to IFNγ can be modulated in the absence of type I IFNs. Multiple mechanisms, thus, exist to calibrate macrophage IFNGR1 abundance, likely permitting the fine tuning of macrophage activation and inflammation.


Online Peptide fractionation using a multiphasic microfluidic liquid chromatography chip improves reproducibility and detection limits for quantitation in discovery and targeted proteomics.
Comprehensive proteomic profiling of biological specimens usually requires multidimensional chromatographic peptide fractionation prior to mass spectrometry. However, this approach can suffer from poor reproducibility because of the lack of standardization and automation of the entire workflow, thus compromising performance of quantitative proteomic investigations. To address these variables we developed an online peptide fractionation system comprising a multiphasic liquid chromatography (LC) chip that integrates reversed phase and strong cation exchange chromatography upstream of the mass spectrometer (MS). We showed superiority of this system for standardizing discovery and targeted proteomic workflows using cancer cell lysates and nondepleted human plasma. Five-step multiphase chip LC MS/MS acquisition showed clear advantages over analyses of unfractionated samples by identifying more peptides, consuming less sample and often improving the lower limits of quantitation, all in highly reproducible, automated, online configuration. We further showed that multiphase chip LC fractionation provided a facile means to detect many N- and C-terminal peptides (including acetylated N terminus) that are challenging to identify in complex tryptic peptide matrices because of less favorable ionization characteristics. Given as much as 95% of peptides were detected in only a single salt fraction from cell lysates we exploited this high reproducibility and coupled it with multiple reaction monitoring on a high-resolution MS instrument (MRM-HR). This approach increased target analyte peak area and improved lower limits of quantitation without negatively influencing variance or bias. Further, we showed a strategy to use multiphase LC chip fractionation LC-MS/MS for ion library generation to integrate with SWATH(TM) data-independent acquisition quantitative workflows. All MS data are available via ProteomeXchange with identifier PXD001464.


Plasma chromogranin A in patients with autoimmune chronic atrophic gastritis, enterochromaffin-like cell lesions and gastric carcinoids.
In atrophic body gastritis (ABG) chronic hypergastrinaemia stimulates enterochromaffin-like (ECL) cell proliferation with development of cell hyperplasia, dysplasia and possibly type-1 gastric carcinoids. As circulating chromogranin A (CgA) levels are a marker of neuroendocrine tumours, we evaluated the clinical usefulness of CgA assay in ABG patients to detect those with carcinoids. Plasma CgA levels were measured using a commercial ELISA in 45 healthy volunteers, nine patients with type-1 gastric carcinoids and 43 consecutive ABG patients (21 without and 22 with ECL cell hyperplasia/dysplasia). CgA levels were significantly higher in ABG patients with and without gastric carcinoids than in healthy subjects (P < 0.001). The highest values occurred in patients with carcinoids (median (interquartile range): 58.1 (44.5-65.3) U/l) and with ECL cell hyperplasia/dysplasia (35.5 (31.8-48.65) U/l) but there were no significant differences in CgA among the various subgroups of ABG patients classified according to ECL cell status. Nevertheless, in ABG patients without carcinoids CgA values correlated with the presence and severity of ECL cell lesions (r(s) = 0.428, P < 0.01). The sensitivity and specificity of the CgA assay in identifying patients with carcinoids were 100 and 23% respectively. CgA plasma levels reflect the histological degree of ECL cell lesions in patients with ABG but the assay specificity is too low to detect among these patients those with gastric carcinoids.


Arcuate nucleus-specific leptin receptor gene therapy attenuates the obesity phenotype of Koletsky (fa(k)/fa(k)) rats.
Leptin signaling in the hypothalamic arcuate nucleus (ARC) is hypothesized to play an important role in energy homeostasis. To investigate whether leptin signaling limited to this brain area is sufficient to reduce food intake and body weight, we used adenoviral gene therapy to express the signaling isoform of the leptin receptor, lepr(b), in the ARC of leptin receptor-deficient Koletsky (fa(k)/fa(k)) rats. Successful expression of adenovirus containing lepr(b) (Ad-lepr(b)) selectively in the ARC was documented by in situ hybridization. Using real-time PCR, we further demonstrated that bilateral microinjection of Ad-lepr(b) into the ARC restored low hypothalamic levels of lepr(b) mRNA to values approximating those of wild-type (Fa(k)/Fa(k)) controls. Restored leptin receptor expression in the ARC reduced both mean daily food intake (by 13%) and body weight gain (by 33%) and increased hypothalamic proopiomelanocortin mRNA by 65% while decreasing neuropeptide Y mRNA levels by 30%, relative to fa(k)/fa(k) rats injected with a control adenovirus (Ad-lacZ) (P < 0.05 for each comparison). In contrast, Ad-lepr(b) delivery to either the lateral hypothalamic area of fa(k)/fa(k) rats or to the ARC of wild-type Fa(k)/Fa(k) rats had no effect on any of these parameters. These findings collectively support the hypothesis that leptin receptor signaling in the ARC is sufficient to mediate major effects of leptin on long-term energy homeostasis. Adenoviral gene therapy is thus a viable strategy with which to study the physiological importance of specific molecules acting in discrete brain areas.


Latency Compensated Visual-Inertial Odometry for Agile Autonomous Flight.
In visual-inertial odometry (VIO), inertial measurement unit (IMU) dead reckoning acts as the dynamic model for flight vehicles while camera vision extracts information about the surrounding environment and determines features or points of interest. With these sensors, the most widely used algorithm for estimating vehicle and feature states for VIO is an extended Kalman filter (EKF). The design of the standard EKF does not inherently allow for time offsets between the timestamps of the IMU and vision data. In fact, sensor-related delays that arise in various realistic conditions are at least partially unknown parameters. A lack of compensation for unknown parameters often leads to a serious impact on the accuracy of VIO systems and systems like them. To compensate for the uncertainties of the unknown time delays, this study incorporates parameter estimation into feature initialization and state estimation. Moreover, computing cross-covariance and estimating delays in online temporal calibration correct residual, Jacobian, and covariance. Results from flight dataset testing validate the improved accuracy of VIO employing latency compensated filtering frameworks. The insights and methods proposed here are ultimately useful in any estimation problem (e.g., multi-sensor fusion scenarios) where compensation for partially unknown time delays can enhance performance.


A simple rule for the evolution of contingent cooperation in large groups.
Humans cooperate in large groups of unrelated individuals, and many authors have argued that such cooperation is sustained by contingent reward and punishment. However, such sanctioning systems can also stabilize a wide range of behaviours, including mutually deleterious behaviours. Moreover, it is very likely that large-scale cooperation is derived in the human lineage. Thus, understanding the evolution of mutually beneficial cooperative behaviour requires knowledge of when strategies that support such behaviour can increase when rare. Here, we derive a simple formula that gives the relatedness necessary for contingent cooperation in n-person iterated games to increase when rare. This rule applies to a wide range of pay-off functions and assumes that the strategies supporting cooperation are based on the presence of a threshold fraction of cooperators. This rule suggests that modest levels of relatedness are sufficient for invasion by strategies that make cooperation contingent on previous cooperation by a small fraction of group members. In contrast, only high levels of relatedness allow the invasion by strategies that require near universal cooperation. In order to derive this formula, we introduce a novel methodology for studying evolution in group structured populations including local and global group-size regulation and fluctuations in group size.


Impact of the 1993 CDC surveillance definition of AIDS in Texas, 1991-1994.
In 1993, the Centers for Disease Control and Prevention (CDC) expanded the surveillance case definition of AIDS to include all individuals showing immune suppression, measured by absolute CD4 lymphocyte count or CD4/CD8 ratio, as well as those presenting with the 23 clinical conditions from previous case definitions or with three new indicator conditions (pulmonary tuberculosis, recurrent pneumonia, and invasive cervical cancer). The greatest impact of the expanded definition is the dramatic increase in cases reported in 1993, largely due to a backlog of 1992-diagnosed cases that meet the new criteria. Texas reported a 136% increase in cases in 1993 compared with 1992. Despite a drop between 1993 and 1994, the number of reported cases increased 75% overall between 1992 and 1994. Most cases reported in 1993 and 1994 fall under the 1993 expanded surveillance case definition (59% and 60%, respectively). More than 90% of all cases reported under the new definition are classified under immunologic criteria. The Department of Health and Human Services (DHHS) Public Health Region 4 was the most affected, with a 213% increase in reported cases between 1992 and 1993, and a 163% increase between years 1992 and 1994. In Texas, the new definition shows the greatest impact among women, blacks, and "other" ethnic groups, persons 20 through 29 years of age, and cases attributable to heterosexual contact. From 1992 to 1993, cases reported in Texas increased 161% among females, 171% among blacks, 219% among persons 15 through 19 years old, and 189% among cases citing heterosexual contact. The new definition has successfully identified population subgroups affected most recently by the epidemic. The inflated growth in cases caused by the expanded case definition carries implications for program administrators and health care providers in both allocating resources and targeting prevention efforts.


[Effectiveness and pharmacodynamics of tobanum in patients with paroxysmal reciprocal atrioventricular tachycardia].
The antiarrhythmic effects and pharmacodynamics of tobanum were evaluated in 28 patients wist paroxysms of reciprocal atrioventricular tachycardia, by using transesophageal cardiac pacing. The agent given in a single dose of 10 mg produced antiarrhythmic effects in 70% patients with paroxysms of atrioventricular nodal tachycardia and in 40% with atrioventricular tachycardia in the presence of the Wolff--Parkinson-White syndrome. After administration of the drug, its antiarrhythmic effect occurred on an average 1.5 h and retained for 27 h.


In utero PCP exposure alters oligodendrocyte differentiation and myelination in developing rat frontal cortex.
Several neurodevelopmental disorders, including schizophrenia, autism, ADD/ADHD and dyslexia are believed to originate during gestation and involve white matter abnormalities. Modulation of glutamate environments and glutamate receptors has also been implicated in alteration of oligodendrocytes, the myelin forming cells of the CNS. To begin to understand how modulation of the glutamate system affects the maturation of oligodendrocytes, developing rats were subjected to prenatal blockade of the NMDA receptor with phencyclidine (PCP). Oligodendrocyte development and differentiation were then examined postnatally by measuring markers for early, middle and late stage cells. The results indicate that, while the level of marker proteins for neurons and astrocytes remains the same, early oligodendrocyte progenitor cell markers are decreased in rat brains prenatally exposed to PCP. Labeling of cells of intermediate, immature cell stages is elevated. Late stage markers for myelinating oligodendrocytes are subsequently decreased. These data suggest that prenatal NMDA receptor blockade reduces the level of progenitors and that the surviving cells are arrested at an immature stage. This premature arrest appears to result in fewer fully differentiated, mature oligodendrocytes that are capable of producing myelin. These results have interesting implications for the role of glutamate and glutamate receptors in white matter abnormalities in neurodevelopmental disorders.


Maternal glutaric acidemia, type I identified by newborn screening.
We report two women with glutaric acidemia type I in whom the diagnosis was unsuspected until a low carnitine level was found in their newborn children. Both mothers had low carnitine in plasma. In the first, organic acid analysis was only done after fibroblast studies revealed normal carnitine uptake. Having learned from the first family, organic acid analysis was done immediately in the mother of family 2. In both, the plasma acylcarnitine profile was normal but both excreted the metabolites typical of their disorder. One of the women was a compound heterozygote for distinct mutations in the glutaric acid dehydrogenase gene, whereas the second was either homozygous or hemizygous for a mutation in Exon 6 of the gene.


Stretch-induced stress fiber remodeling and the activations of JNK and ERK depend on mechanical strain rate, but not FAK.
Cells within tissues are subjected to mechanical forces caused by extracellular matrix deformation. Cells sense and dynamically respond to stretching of the matrix by reorienting their actin stress fibers and by activating intracellular signaling proteins, including focal adhesion kinase (FAK) and the mitogen-activated proteins kinases (MAPKs). Theoretical analyses predict that stress fibers can relax perturbations in tension depending on the rate of matrix strain. Thus, we hypothesized stress fiber organization and MAPK activities are altered to an extent dependent on stretch frequency. Bovine aortic endothelial cells and human osteosarcoma cells expressing GFP-actin were cultured on elastic membranes and subjected to various patterns of stretch. Cyclic stretching resulted in strain rate-dependent increases in stress fiber alignment, cell retraction, and the phosphorylation of the MAPKs JNK, ERK and p38. Transient step changes in strain rate caused proportional transient changes in the levels of JNK and ERK phosphorylations without affecting stress fiber organization. Disrupting stress fiber contractile function with cytochalasin D or Y27632 decreased the levels of JNK and ERK phosphorylation. Previous studies indicate that FAK is required for stretch-induced cell alignment and MAPK activations. However, cyclic uniaxial stretching induced stress fiber alignment and the phosphorylation of JNK, ERK and p38 to comparable levels in FAK-null and FAK-expressing mouse embryonic fibroblasts. These results indicate that cyclic stretch-induced stress fiber alignment, cell retraction, and MAPK activations occur as a consequence of perturbations in fiber strain. These findings thus shed new light into the roles of stress fiber relaxation and reorganization in maintenance of tensional homeostasis in a dynamic mechanical environment.


Parent-Adolescent Relationship Qualities, Internal Working Models, and Styles as Predictors of Adolescents' Observed Interactions with Friends.
This study examined how current parent-adolescent relationship qualities and adolescents' representations of relationships with parents were related to friendship interactions in 200 adolescent-close friend dyads. Adolescents and friends were observed discussing problems during a series of structured tasks. Negative interactions with mothers were significantly related to adolescents' greater conflict with friends, poorer focus on tasks, and poorer communication skills. Security of working models (as assessed by interview) was significantly associated with qualities of friendship interactions, whereas security of attachment styles (as assessed by questionnaire) was not. More dismissing (vs. secure) working models were associated with poorer focus on problem discussions and weaker communication skills with friends, even after accounting for gender differences and current parent-adolescent relationship qualities. We discuss possible mechanisms for the observed links between dimensions of parent-adolescent relationships and friendships. We also consider methodological and conceptual differences between working model and style measures of attachment representations.


German language questionnaires for assessing implementation constructs and outcomes of psychosocial and health-related interventions: a systematic review.
Over the past years, implementation science has gained more and more importance in German-speaking countries. Reliable and valid questionnaires are needed for evaluating the implementation of evidence-based practices. On an international level, several initiatives focused on the identification of questionnaires used in English-speaking countries but limited their search processes to mental health and public health settings. Our aim was to identify questionnaires used in German-speaking countries measuring the implementation of interventions in public health and health care settings in general and to assess their psychometric properties. We searched five different bibliographic databases (from 1985 to August 2017) and used several other search strategies (e.g., reference lists, forward citation) to obtain our data. We assessed the instruments, which were identified in an independent dual review process, using 12 psychometric rating criteria. Finally, we mapped the instruments' scales and subscales in regard to the constructs of the Consolidated Framework for Implementation Research (CFIR) and the Implementation Outcome Framework (IOF). We identified 31 unique instruments available for the assessment of implementation science constructs. Hospitals and other health care settings were the ones most often investigated (23 instruments), while education and childcare settings, workplace settings, and community settings lacked published instruments. Internal consistency, face and content validity, usability, and structural validity were the aspects most often described. However, most studies did not report on test-retest reliability, known-groups validity, predictive criterion validity, or responsiveness. Overall, the majority of studies did not reveal high-quality instruments, especially regarding the psychometric criteria internal consistency, structural validity, and criterion validity. In addition, we seldom detected instruments operationalizing the CFIR domains intervention characteristics, outer setting, and process, and the IOF constructs adoption, fidelity, penetration, and sustainability. Overall, a sustained and continuous effort is needed to improve the reliability and validity of existing instruments to new ones. Instruments applicable to the assessment of implementation constructs in public health and community settings are urgently needed. The systematic review protocol was registered in PROSPERO on October 19, 2017, under the following number: CRD42017075208 .


The intergenerational transmission of health-risk behaviors: adolescent lifestyles and gender moderating effects.
The present longitudinal study of 330 adolescents used structural equation models to investigate whether (1) health-risk lifestyles exist among adults and adolescents, (2) parents' health-risk behaviors influence adolescents' health-risk behaviors, and (3) intergenerational transmission occurs by way of a health-risk lifestyle, by direct transmission of specific behaviors, or through both mechanisms. To address these questions, we estimated several models. The findings were generally supportive of the expectations. Results of single factor measurement models provided modest evidence for the existence of an underlying health-risk lifestyle factor among parents and adolescents. Results of structural equation models also demonstrated that parents' health-risk behaviors were transmitted to adolescents both at the lifestyle factor level and the unique component level. These associations prevailed even after controlling for family social status. However, parents' health-risk lifestyles mediated the effect of family social status on adolescents' lifestyles, net of the direct effect of family social status on adolescents' lifestyle. In these two-parent families, the effects of parents' health-risk lifestyles on adolescents seems to have gender symmetry. The findings of the separate models for boys and girls demonstrated that (1) fathers' health-risk lifestyle affected only boys' health-risk lifestyle, whereas (2) mother's health-risk lifestyle affected only girls' health-risk lifestyle. A similar gender moderating effect was not found for specific health-risk behaviors. Implications of these findings for future research and theoretical development are discussed.


Interventions for smoking cessation in hospitalised patients.
An admission to hospital provides an opportunity to help people stop smoking. Individuals may be more open to help at a time of perceived vulnerability, and may find it easier to quit in an environment where smoking is restricted or prohibited. Initiating smoking cessation services during hospitalisation may help more people to make and sustain a quit attempt. To determine the effectiveness of interventions for smoking cessation that are initiated for hospitalised patients. We searched the Cochrane Tobacco Addiction Group register which includes papers identified from CENTRAL, MEDLINE, EMBASE and PSYCINFO in January 2007, and CINAHL in August 2006 for studies of interventions for smoking cessation in hospitalised patients, using terms including (hospital and patient*) or hospitali* or inpatient* or admission* or admitted. Randomized and quasi-randomized trials of behavioural, pharmacological or multicomponent interventions to help patients stop smoking, conducted with hospitalised patients who were current smokers or recent quitters (defined as having quit more than one month before hospital admission). The intervention had to start in the hospital but could continue after hospital discharge. We excluded studies of patients admitted for psychiatric disorders or substance abuse, studies that did not report abstinence rates and studies with follow up of less than six months. Two authors extracted data independently for each paper, with disagreements resolved by consensus. Thirty-three trials met the inclusion criteria. Intensive counselling interventions that began during the hospital stay and continued with supportive contacts for at least one month after discharge increased smoking cessation rates after discharge (Odds Ratio (OR) 1.65, 95% confidence interval (CI) 1.44 to 1.90; 17 trials). No statistically significant benefit was found for less intensive counselling interventions. The one study that tested a single brief (<=15 minutes) in-hospital intervention did not find it to be effective (OR 1.16, 95% CI 0.80 to 1.67). Counselling of longer duration during the hospital stay was not associated with a higher quit rate (OR 1.08, 95% CI 0.89 to 1.29, eight trials). Even counselling that began in the hospital but had less than one month of supportive contact after discharge did not show significant benefit (OR 1.09, 95% CI 0.91 to 1.31, six trials). Adding nicotine replacement therapy (NRT) did not produce a statistically significant increase in cessation over what was achieved by intensive counselling alone (OR 1.47, 95% CI 0.92 to 2.35, five studies). The one study that tested the effect of adding bupropion to intensive counselling had a similar nonsignificant effect (OR 1.56, 95% CI 0.79 to 3.06). A similar pattern of results was observed in smokers admitted to hospital because of cardiovascular disease (CVD). In this subgroup, intensive intervention with follow-up support increased the odds of smoking cessation (OR 1.81, 95% CI 1.54 to 2.15, 11 trials), but less intensive interventions did not. One trial of intensive intervention including counselling and pharmacotherapy for smokers admitted with CVD assessed clinical and health care utilization endpoints, and found significant reductions in all-cause mortality and hospital readmission rates over a two-year follow-up period. High intensity behavioural interventions that begin during a hospital stay and include at least one month of supportive contact after discharge promote smoking cessation among hospitalised patients. These interventions are effective regardless of the patient's admitting diagnosis. lnterventions of lower intensity or shorter duration have not been shown to be effective in this setting. There is insufficient direct evidence to conclude that adding NRT or bupropion to intensive counselling increases cessation rates over what is achieved by counselling alone, but the evidence of benefit for NRT has strengthened in this update and the point estimates are compatible with research in other settings showing that NRT and bupropion are effective.


[Anatomy of the vegetative organs of two species of Atriplex (Chenopodiaceae) from Venezuela].
In Venezuela, Atriplex is represented by A. cristata and A. oestophora, the latter being endemic; they inhabit coastal areas with high temperatures, high solar radiation and sandy soils with high salt content. This work aimed to provide information to facilitate and clarify these species taxonomic delimitation, throughout the study of the anatomy of their vegetative organs; this may also clarify our understanding of their adaptability to soil and climatic conditions prevailing in areas they inhabit. The plant material was collected from at least three individuals of each species in Punta Taima Taima and Capatárida, Falcon. Segments of roots, located near the neck and towards the apex, apical, middle and basal internodes of stems, were taken; and of leaves, located in the middle portion of plants. This material was fixed in FAA (formaldehyde, acetic acid, 70% ethanol) until processing. Semipermanent and permanent microscope slides were prepared with transverse or longitudinal sections, made using a razor (free-hand) or a rotation microtome, in this latter case, after paraffin embedding; besides, additional plates were mounted with portions of leaf epidermis, obtained by the maceration technique. The sections were stained with aqueous toluidine blue (1%) or safranin-fast-green, and mounted in water-glycerin or in Canada balsam. In order to calculate the vulnerability index, the vessel diameter in the vascular rings of roots, as well as their density, were quantified. Our results revealed structural features in the different organs, that resulted of taxonomic value and allowed the distinction of the species: in the leaf, the presence of aquifer tissue, the number of vascular bundles and their organization in the midrib, and the collenchyma differentiation in this part of the leaf; in the roots, the xylem and phloem arrangement in the growth rings, the nature of conjunctive tissue, and the presence of included phloem in one species. In addition, the species showed typical anatomical features of halophytes and xerophytes, such as: high density of trichomes on leaves and young stems which act as salt secreting glands, abundant sclerenchyma in stems and roots, water storage tissue and Kranz anatomy in leaves, narrow cortical region in young roots, presence of cambial variants in stems and roots, as well as short and narrow xylem vessels. Vulnerability index calculations indicated that both species tend to assure conduction but not the efficiency of the system. Atriplex species have anatomical characters which facilitate their adaptation to the special conditions prevailing in their habitats and that may be used for taxonomic delimitation.


Podocyte-Specific Loss of Krüppel-Like Factor 6 Increases Mitochondrial Injury in Diabetic Kidney Disease.
Mitochondrial injury is uniformly observed in several murine models as well as in individuals with diabetic kidney disease (DKD). Although emerging evidence has highlighted the role of key transcriptional regulators in mitochondrial biogenesis, little is known about the regulation of mitochondrial cytochrome c oxidase assembly in the podocyte under diabetic conditions. We recently reported a critical role of the zinc finger Krüppel-like factor 6 (KLF6) in maintaining mitochondrial function and preventing apoptosis in a proteinuric murine model. In this study, we report that podocyte-specific knockdown of Klf6 increased the susceptibility to streptozotocin-induced DKD in the resistant C57BL/6 mouse strain. We observed that the loss of KLF6 in podocytes reduced the expression of synthesis of cytochrome c oxidase 2 with resultant increased mitochondrial injury, leading to activation of the intrinsic apoptotic pathway under diabetic conditions. Conversely, mitochondrial injury and apoptosis were significantly attenuated with overexpression of KLF6 in cultured human podocytes under hyperglycemic conditions. Finally, we observed a significant reduction in glomerular and podocyte-specific expression of KLF6 in human kidney biopsies with progression of DKD. Collectively, these data suggest that podocyte-specific KLF6 is critical to preventing mitochondrial injury and apoptosis under diabetic conditions.


Effects of powdered whole grapefruit and metoclopramide on the pharmacokinetics of cyclosporine in dogs.
To determine whether oral administration of metoclopramide or a commercially available powdered whole grapefruit (PWG) nutraceutical in combination with cyclosporine enhances systemic availability of cyclosporine in dogs. 8 healthy mixed-breed dogs in part 1 and 6 of these 8 dogs in part 2. Cyclosporine pharmacokinetics were determined over the course of 24 hours after oral administration of cyclosporine (5 mg/kg) alone, cyclosporine with metoclopramide (0.3 to 0.5 mg/kg), cyclosporine with 2 g of PWG, or cyclosporine combined with both metoclopramide and 2 g of PWG by use of a Latin square crossover study with a 14-day washout period between treatments. Sixty days later, 6 of the 8 dogs were given 10 g of PWG followed by cyclosporine, and pharmacokinetic parameters were compared with those previously obtained after administration of cyclosporine alone. Although metoclopramide or coadministration of metoclopramide and 2 g of PWG had no effect on the pharmacokinetic parameters of cyclosporine, compared with results for cyclosporine alone, the higher (10-g) dose of PWG resulted in 29% faster mean time to maximal plasma cyclosporine concentration, 54% larger area under the curve, and 38% lower apparent oral clearance. Adjustment of the cyclosporine dose may not be needed when metoclopramide is coadministered orally to prevent common adverse effects of cyclosporine. Powdered whole grapefruit has the potential to reduce the required orally administered dose of cyclosporine but only when PWG is used in an amount (at least 10 g) that is currently not cost-effective.


Role of antizyme in degradation of ornithine decarboxylase in HTC cells.
A good correlation was observed between the reciprocal of the half-life of ornithine decarboxylase (ODC) activity in the presence of cycloheximide and the relative amount of ODC-antizyme complex to total ODC (free ODC plus complexed ODC) activity in HTC cells examined at various times after cell dilution or change of medium. Pretreatment of cells with putrescine increased the relative amount of ODC-antizyme complex and decreased the half-life of ODC decay. These results suggested that antizyme plays a key role in ODC degradation.


Prediction of Response to Neoadjuvant Chemotherapy and Radiation Therapy with Baseline and Restaging 18F-FDG PET Imaging Biomarkers in Patients with Esophageal Cancer.
Purpose To assess the value of baseline and restaging fluorine 18 (18F) fluorodeoxyglucose (FDG) positron emission tomography (PET) radiomics in predicting pathologic complete response to neoadjuvant chemotherapy and radiation therapy (NCRT) in patients with locally advanced esophageal cancer. Materials and Methods In this retrospective study, 73 patients with histologic analysis-confirmed T1/N1-3/M0 or T2-4a/N0-3/M0 esophageal cancer were treated with NCRT followed by surgery (Chemoradiotherapy for Esophageal Cancer followed by Surgery Study regimen) between October 2014 and August 2017. Clinical variables and radiomic features from baseline and restaging 18F-FDG PET were selected by univariable logistic regression and least absolute shrinkage and selection operator. The selected variables were used to fit a multivariable logistic regression model, which was internally validated by using bootstrap resampling with 20 000 replicates. The performance of this model was compared with reference prediction models composed of maximum standardized uptake value metrics, clinical variables, and maximum standardized uptake value at baseline NCRT radiomic features. Outcome was defined as complete versus incomplete pathologic response (tumor regression grade 1 vs 2-5 according to the Mandard classification). Results Pathologic response was complete in 16 patients (21.9%) and incomplete in 57 patients (78.1%). A prediction model combining clinical T-stage and restaging NCRT (post-NCRT) joint maximum (quantifying image orderliness) yielded an optimism-corrected area under the receiver operating characteristics curve of 0.81. Post-NCRT joint maximum was replaceable with five other redundant post-NCRT radiomic features that provided equal model performance. All reference prediction models exhibited substantially lower discriminatory accuracy. Conclusion The combination of clinical T-staging and quantitative assessment of post-NCRT 18F-FDG PET orderliness (joint maximum) provided high discriminatory accuracy in predicting pathologic complete response in patients with esophageal cancer. © RSNA, 2018 Online supplemental material is available for this article.


Ivabradine in chronic stable angina: Effects by and beyond heart rate reduction.
Heart rate plays a major role in myocardial ischemia. A high heart rate increases myocardial performance and oxygen demand and reduces diastolic time. Ivabradine reduces heart rate by inhibiting the If current of sinoatrial-node cells. In contrast to beta-blockers, ivabradine has no negative inotropic and lusitropic effect for a comparable heart rate reduction. Consequently, diastolic duration is increased with ivabradine compared to beta-blockers. This has potential consequences on coronary blood flow since compression of the vasculature by the surrounding myocardium during systole impedes flow and coronary blood flow is mainly diastolic. Moreover, ivabradine does not unmask alpha-adrenergic vasoconstriction and, unlike beta-blockers, maintains coronary dilation during exercise. In comparison with beta-blockers, ivabradine increases coronary flow reserve and collateral perfusion promoting the development of coronary collaterals. Ivabradine attenuates myocardial ischemia and its consequences even in the absence of heart rate reduction, possibly through reduced formation of reactive oxygen species. In conclusion, ivabradine differs from other anti-anginal agents by improving coronary blood flow and by additional pleiotropic effects. These properties make ivabradine an effective anti-anginal and anti-ischemic agent for the treatment of patients with coronary artery disease.


Computer-aided interpretation of SPECT images of the brain using an MRI-derived 3D neuro-anatomical atlas.
Nuclear medicine images have comparatively poor spatial resolution, making it difficult to relate the functional information which they contain to precise anatomical structures. A 3D neuro-anatomical atlas has been generated from the MRI data set of a normal, healthy volunteer to assist in the interpretation of nuclear medicine scans of the brain. Region growing and edge-detection techniques were used to semi-automatically segment the data set into the major tissue types within the brain. The atlas was then labelled interactively by marking points on each 2D slice. Anatomical structures useful in the interpretation of SPECT images were labelled. Additional, more detailed information corresponding to these structures is provided via an interactive index which allows access to images, diagrams and explanations. Registration of patient SPECT studies with the atlas is accomplished by using the position of the skull vertex and four external fiducial markers attached to the skin surface. The 3D coordinates determined from these points are used to calculate the transformation required to rotate, scale and translate the SPECT data, in 3D, to match the atlas. Corresponding 2D slices from the two 3D data sets are then displayed side-by-side on a computer screen. A cursor linking the two images allows the delineation of regions of interest (ROIs) in the SPECT scan based on anatomical structures identified from the atlas. Conversely regions of abnormal isotope distribution in the SPECT image can be localized by reference to corresponding structures in the atlas.


C4d deposition in early renal allograft protocol biopsies.
Deposition of the complement protein C4d in renal allograft biopsies obtained during graft dysfunction and rejection has been proposed to be a sensitive marker of antibody-mediated acute rejection. To determine the diagnostic specificity of C4d deposition, it is important to study biopsies from allografts with no evidence of dysfunction. In this study, we examined C4d deposition in protocol biopsies obtained irrespective of clinical status. Immunohistochemistry for C4d was performed on routine protocol biopsies preimplantation and on day 7 posttransplantation from 48 unselected renal allografts. Serum samples obtained up to 1 month after transplantation were assayed for donor-reactive antibodies (DRA). Results were correlated with histopathology and clinical outcome measures. Diffuse C4d deposition was detected in the peritubular capillaries of 6 of 48 (13%) biopsies. C4d deposition was present in 5 of 15 (33%) biopsies that showed acute rejection (Banff 97, category 4) but only in 1 of 33 (3%) biopsies with no rejection (P=0.003, 97% specificity). Posttransplant DRAs were detected in 21 of 48 (44%) patients. All five recipients with C4d deposition and rejection had posttransplant DRA; the recipient whose biopsy showed C4d positivity, but not rejection, did not have detectable DRA. C4d deposition was not treated with plasmapheresis or intravenous immunoglobulin and was not associated with poor posttransplant graft outcome at 1-year follow-up. Our results show that in early posttransplant protocol biopsies, C4d is a specific marker for the presence of humoral rejection, as indicated by its association with DRA and acute histologic rejection.


Use of sodium bicarbonate to treat tricyclic antidepressant-induced arrhythmias in a patient with alkalosis.
Sodium bicarbonate has been recommended for the treatment of arrhythmias induced by tricyclic antidepressants. It is unclear, however, whether this therapy is effective only in the presence of acidosis. A case is presented in which there was an immediate response to sodium bicarbonate in three episodes of ventricular tachycardia despite the presence of alkalosis on two of the three occasions. Given the poor response to conventional therapy of arrhythmias induced by tricyclic antidepressants the use of sodium bicarbonate may be reasonable even in the presence of alkalosis. However, in the presence of pre-existing respiratory or metabolic alkalosis, such therapy is not without risk, and it is suggested that it be reserved for life-threatening situations when the arrhythmia has failed to respond to hyperventilation or antiarrhythmics or both.


Ultrastructural immunocytochemical distribution of VIP-like immunoreactivity in dog ileum.
The electron-immunocytochemical protein A-gold technique was employed to study the subcellular localization of vasoactive intestinal polypeptide-like material in dog ileum. The vasoactive intestinal polypeptide-like immunoreactivity was found within a population of large granular vesicles similar in structure in nerve varicosities of the myenteric plexus, the deep muscular plexus, the submucous plexus, the longitudinal muscular layer and the mucosa; none was found in nerve cell bodies. In the myenteric plexus, submucous plexus, the mucosa and the longitudinal muscular layer, varicosities containing similar large granular vesicles consistently remained unstained suggesting that within these plexuses morphologically indistinguishable by our technique large granular vesicles are not necessarily biochemically identical. In the deep muscular plexus, nearly all varicosities with large granular vesicles contained immunoreactivity for vasoactive intestinal polypeptide, but these varicosities often contained a few unstained large granular vesicles. This suggests that vasoactive intestinal polypeptide may share the same varicosity or the same vesicle with other neuropeptides present in this plexus (e.g., substance P or enkephalins) and that this plexus is a site where vasoactive intestinal polypeptide exerts its control over motility.


Burnout and work engagement: a thorough investigation of the independency of both constructs.
This study among 528 South African employees working in the construction industry examined the dimensionality of burnout and work engagement, using the Maslach Burnout Inventory-General Survey, the Oldenburg Burnout Inventory, and the Utrecht Work Engagement Scale. On the basis of the literature, we predicted that cynicism and dedication are opposite ends of one underlying attitude dimension (called "identification"), and that exhaustion and vigor are opposite ends of one "energy" dimension. Confirmatory factor analyses showed that while the attitude constructs represent opposite ends of one continuum, the energy constructs do not-although they are highly correlated. These findings are also supported by the pattern of relationships between burnout and work engagement on the one hand, and predictors (i.e., work pressure, autonomy) and outcomes (i.e., organizational commitment, mental health) on the other hand. Implications for the measurement and conceptualization of burnout and work engagement are discussed.


Metabolic engineering--a genetic toolbox for small molecule organic synthesis.
Metabolic engineering is an emerging field that exploits biosynthetic machinery as a means to genetically design small molecule production within heterologous host organisms. Molecular design and synthesis with biological tools has lagged behind total synthesis technology for about seventy-five years and owes its existence to relatively new molecular biology techniques. Here the field of metabolic engineering is explained as a comparison to total organic synthesis, including the sequence of scientific events leading up to successful implementation and future goals in the field. It is expected that metabolic engineering will take a place alongside traditional organic synthesis as a powerful means to design and create small organic molecules.


Urinary potassium is a clinically useful test to detect a poor quality diet.
Poor eating habits, a strong predictor of health outcomes, are not objectively assessed in routine clinical practice. In this study, we evaluated the use of urinary potassium (K(+)) as a means to identify people consuming a poor quality diet. Consecutive patients with kidney stones (n = 220), aged 18-50 y, from a population-based lithotripsy unit, collected a single 24-h urine sample to assess urinary K(+). They also completed a FFQ to derive the recommended foods score (RFS), an index of overall diet quality, and had their blood pressure, heart rate, weight, and height measured. Urinary K(+) was related positively with the intake of recommended food items, including vegetables, fruit, whole grains, low-fat dairy products, fish and poultry, and wine and negatively to those not recommended by current dietary guidelines, including red meat, fast food, and high-energy drinks. Urinary K(+) was also correlated with the RFS (r = 0.226; P < 0.001). Using a receiver operating characteristic curve, K(+) excretion values below the gender-specific median (men, 60 mmol/d; women, 41 mmol/d) were identified as the optimal cutoff values for a poor quality diet, indicated by the RFS. Higher urinary K(+) was inversely related to adjusted BMI (P-trend = 0.03), diastolic blood pressure (P-trend = 0.04) and heart rate (P-trend = 0.006), after controlling for potential confounders. Urinary K(+) provides a summary measure of diet quality, is significantly related to BMI, blood pressure, and heart rate, and may be useful clinically to detect poor dietary habits and monitor response to dietary interventions.


NOTES spin-off for the therapeutic gastroenterologist: natural orifice surgery.
Natural orifice transluminal endoscopic surgery (NOTES) has ushered in a new era in flexible endoscopy. Over the past decade, modest advances have been made in addressing the fundamental challenges of NOTES surgery including transluminal access and closure techniques, and advancement of NOTES-specific technology. Despite these encouraging initial efforts significant obstacles to widespread acceptance of NOTES as a surgical option persist. Moreover, due to the well-documented safety and efficacy of laparoscopic techniques, the question remains as to the best candidate NOTES procedure. Presently, interest has shifted from true NOTES to hybrid procedures and single incision laparoscopic surgery, due to the lure of more immediate success. Additionally, there is also a growing awareness of the potential applications of natural orifice surgery techniques to the present field of therapeutic endoscopy. Research into transluminal access and closure has born several techniques and devices that are now being explored in endoscopic procedures such as full-thickness resection, endoscopic myotomy, direct endoscopic pancreatic necrosectomy and bariatric endoscopy. Such NOTES "spin-off" procedures are expanding the armamentarium of today's therapeutic endoscopists, and a growing body of literature suggests that they will play a significant role in the evolution of therapeutic endoscopy in the future.


Outcomes After Sleeve Lung Resections Versus Pneumonectomy in the United States.
The current national trends, practice patterns, and outcomes after sleeve resection compared with pneumonectomy in the United States are not known. In addition, whether hospital sleeve-to-pneumonectomy (S:P) ratios are a valid marker of hospital quality is unclear. We describe practice patterns and evaluate the utility of the S:P ratio. We identified all patients (N = 23,964) undergoing sleeve resection (n = 1,713) or pneumonectomy (n = 22,251) in the National Cancer Data Base between 1998 and 2012 at 644 hospitals. We used propensity score matching to compare short-term outcomes and overall survival between pneumonectomy and sleeve resection. We grouped hospitals into S:P ratio quintiles and used multilevel modeling to analyze hospital-level outcomes. There has been a 1% yearly increase in sleeve resection rates, with wide variation in hospital S:P ratios (middle quintile, 1:12; range, 1:38 to 1:3). After propensity score matching, differences in age, clinical T and N stage, and the incidence of main bronchus tumors were negligible among other variables. Sleeve resections were associated with lower 30-day (1.6% vs 5.9%; p < 0.001) and 90-day mortality (4% vs 9.4%; p < 0.001) and improved overall survival. Hospitals with higher S:P ratios were not associated with better risk-adjusted 30-day (7.2% vs 7.4%; p = 0.244) or 90-day mortality (11.9% vs 12.2%; p = 0.308) or same-hospital readmission rates (3.7% vs 4.3%; p = 0.523). Compared with pneumonectomy, sleeve resections are associated with improved short-term outcomes and improved overall survival. However, hospital S:P ratios were not associated with better risk-adjusted outcomes and thus may not be a valid quality measure.


Effect of Cefepime on Neurotoxicity Development in Critically Ill Adults With Renal Dysfunction.
Pharmacodynamic and pathophysiologic changes in critically ill adults receiving cefepime may increase the risk of adverse events. What is the impact of cefepime exposure on neurotoxicity development in critically ill adults with renal dysfunction? Critically ill adults with creatinine clearance < 60 mL/min who received cefepime for ≥ 48 hours between January 1, 2014 and July 31, 2018 were evaluated for cefepime-associated neurotoxicity (CAN) development. Higher- and lower-dose cefepime exposure groups stratified by moderate (≥ 8 g vs < 8 g in first 48 hours) or severe (≥ 4 g vs < 4 g in first 48 hours) renal dysfunction were compared. Between-group comparisons were performed using Fisher exact tests. CAN-free survival was evaluated using Kaplan-Meier curves and log-rank tests. Cefepime total dose in the first 48 hours was greater in the higher-dose cefepime group (3.7 ± 1.6 g vs 7.7 ± 2.2 g; P < .001). Cefepime-associated neurotoxicity occurred infrequently in both lower- (n = 108) and higher-dose (n = 92) cefepime groups (4% vs 10%, OR 2.82, 95% CI, 0.84-9.48, P = .093). The frequencies of cefepime-associated neurotoxicity were similar between lower- and higher-dose cefepime groups when moderate renal dysfunction subgroups were compared (5% vs 7%, OR 1.42, 95% CI, 0.34-5.92, P = .72) and numerically greater in the higher-dose cefepime group in the severe renal dysfunction subgroup (0 vs 16%, P = .064). Times to cefepime-associated neurotoxicity development and resolution were similar between lower- and higher-dose groups. Durations of CAN-free survival were similar between lower- and higher-dose groups. Most patients who developed cefepime-associated neurotoxicity displayed altered mental status (n = 12, 92%). Cefepime-associated neurotoxicity is an uncommon occurrence in critically ill adults. Patients with severe renal dysfunction receiving higher-dose cefepime may be at greater risk of cefepime-associated neurotoxicity, although this requires additional investigation.


Concomitant Non-V600E BRAF and KRAS Mutations in Colorectal Carcinoma by Next-Generation Sequencing: A Distinct Subtype.
The RAS-RAF-MEK-ERK signaling cascade is the most frequently affected signaling pathway in colorectal cancer. BRAFV600E mutations serve as a drug-treatable hotspot and KRAS mutations as a predictor of susceptibility to anti-epidermal growth factor receptor therapy. Concomitant non-V600E BRAF and KRAS mutations may coexist and are rarely reported in the literature. We report a patient of colorectal carcinoma with inguinal lymph node metastases harboring mutations at the KRAS and BRAF non-V600E mutation codon detected by next-generation sequencing with an emphasis on clinical, pathological, and therapeutic implications of the mutation and review of the literature.


Size-dependent toxicity and interactions of calcium oxalate dihydrate crystals on Vero renal epithelial cells.
Urinary crystals in normal and kidney stone patients often have varying sizes; the interaction between renal epithelial cells and COD crystals generated in the tubular fluid could play an initiating role in the pathophysiology of calcium oxalate nephrolithiasis. This study aims to compare the cytotoxicity of micro/nano-calcium oxalate dihydrate (COD) crystals (50 nm, 100 nm, 600 nm, 3 μm, and 10 μm) toward African green monkey renal epithelial (Vero) cells to reveal the mechanism of kidney stone formation at the molecular and cellular levels. Vero cells were exposed to COD crystals of varying sizes at a concentration of 200 μg mL-1 for 6 h. The effects of COD crystals on Vero cell viability, apoptosis rate, and cellular biochemical parameters [lactate dehydrogenase (LDH), superoxide dismutase (SOD), reactive oxygen species (ROS), hyaluronic acid (HA), osteopontin (OPN), and mitochondrial membrane potential (Δψm)] were determined using biochemical and morphological analyses. Vero cell viability and apoptotic rate were closely associated with the size of COD crystals; lower cell viability and higher apoptosis rate were observed in cells exposed to smaller COD crystal size. The expression of SOD, ROS, HA and OPN also changed in a size-dependent manner after exposure to the five different sizes of COD crystals. The area ratio of the (100) face with a high density of Ca2+ ions to the total surface area was also found to influence the severity of cell injury. Cell injury induced by COD crystals was mainly caused by excessive expression of intracellular ROS and reduction of free-radical scavenger SOD. Moreover, binding of large crystals on the cell membrane surface takes more time to cause cell injury than internalized small-sized crystals. The cell death rate was found to be positively correlated with the amount of internalized COD crystals. although the COD toxicity is often disregarded, the size-dependent cytotoxicity of COD crystals toward Vero cells is demonstrated in this study.


The cost of chronic dialysis in multiple myeloma.
The complications and costs of chronic dialysis in 4 patients with renal failure due to multiple myeloma are presented. In three patients the paraprotein responded to chemotherapy though without recovery of renal function. These three patients are alive after 18, 16 and 15 months of dialysis, the other dying after 7 months. Hospital admissions ranged from 26 to 74 days per year with infections accounting for 54 to 87% of admission days, 62.5% of which occurred during the first three months of dialysis treatment, with an incidence of 2.4 to 6.9 admissions episodes per year. An in-house audit of our chronic dialysis patients indicated that treatment of myeloma patients is 5-33% more expensive. The extra cost in such high risk patients is mostly due to the greater number and longer duration of hospital admissions for infection. The other extra costs (in decreasing value) of blood products, antibiotics and chemotherapy are relatively small in comparison to in-patient treatment.


Proteomes of pathogenic Escherichia coli/Shigella group surveyed in their host environments.
Proteomic studies on Shigella dysenteriae, Shigella flexneri, enterohemorrhagic Escherichia coli and uropathogenic E. coli (UPEC) are reviewed. UPEC causes infections in the urogenital tract, whereas the other species colonize and, to varying degrees, invade the intestinal tract. Type III secretion systems used to breach the mucosal barrier by the intestinal pathogens revealed distinct expression patterns in different host environments. Dynamic adaptations to changes in nutrient availability and oxygen were observed, including increased reliance on anaerobic respiration and mixed acid fermentation in vivo. Utilization of carbon and nitrogen resources by the bacteria varied considerably depending on the host model investigated. Shigellae and UPEC adapted to metal ion sequestration in the mammalian host by enhancing expression of various receptors and transporters for iron and zinc. This appears to reflect the preferred intracellular life stage of Shigella spp. and responses of UPEC to high levels of lipocalin and lactotransferrin in the urinary tract.


Bsp1, a fungal CPI motif protein, regulates actin filament capping in endocytosis and cytokinesis.
The capping of barbed filament ends is a fundamental mechanism for actin regulation. Capping protein controls filament growth and actin turnover in cells by binding to the barbed ends of the filaments with high affinity and slow off-rate. The interaction between Capping protein and actin is regulated by capping protein interaction (CPI) motif proteins. We identified a novel CPI motif protein, Bsp1, which is involved in cytokinesis and endocytosis in budding yeast. We demonstrate that Bsp1 is an actin binding protein with a high affinity for capping protein via its CPI motif. In cells, Bsp1 regulates capping protein at endocytic sites and is a major recruiter of capping protein to the cytokinetic actin ring. Lastly, we define Bsp1-related proteins as a distinct fungi-specific CPI protein group. Our results suggest that Bsp1 promotes actin filament capping by the capping protein. This study establishes Bsp1 as a new capping protein regulator and promising candidate to regulate actin networks in fungi.


Trade-off between deer herbivory and nitrogen competition alter grassland forb composition.
Two of the major factors that control the composition of herbaceous plant communities are competition for limiting soil resources and herbivory. We present results from a 14-year full factorial experiment in a tallgrass prairie ecosystem that crossed nitrogen (N) addition with fencing to exclude white-tailed deer, Odocoileus virginianus, from half the plots. Deer presence was associated with only modest decreases in aboveground plant biomass (14% decrease; -45 ± 19 g m-2) with no interaction with N addition. N addition at 5.44 and 9.52 g N m-2 year-1 led to increases in biomass. There were weak increases in species richness associated with deer presence, but only for no or low added N (1 and 2 g N m-2 year-1). However, the presence of deer greatly impacted the abundances of some of the dominant perennial forb species, but not the dominant grasses. Deer presence increased the abundance of the forb Artemisia ludoviciana by 34 ± 12 SE g m-2 (94%) and decreased the forb Solidago rigida by 32 ± 13 SE g m-2 (79%). We suggest that these changes may have resulted from trade-off in plant competitive ability for soil N versus resistance to deer herbivory. Field observations suggest deer acted as florivores, mainly consuming the flowers of susceptible forb species. The preferential consumption of flowers of forbs that seem to be superior N competitors appears to create an axis of interspecific niche differentiation. The overpopulation of white-tailed deer in many tallgrass reserves likely structures the abundance of forb species.


Fidelity of RNA templated end-joining by chlorella virus DNA ligase and a novel iLock assay with improved direct RNA detection accuracy.
Ligation-based nucleic acid detection methods are primarily limited to DNA, since they exhibit poor performance on RNA. This is attributed to reduced end-joining efficiency and/or fidelity of ligases. Interestingly, chlorella virus DNA ligase (PBCV-1 DNA ligase) has recently been shown to possess high RNA-templated DNA end-joining activity; however, its fidelity has not yet been systematically evaluated. Herein, we characterized PBCV-1 ligase for its RNA-templated end-joining fidelity at single base mismatches in 3' and 5' DNA probe termini and found an overall limited end-joining fidelity. To improve the specificity in PBCV-1 ligase-driven RNA detection assays, we utilized structure-specific 5' exonucleolytic activity of Thermus aquaticus DNA polymerase, used in the invader assay. In the iLock (invader padLock) probe assay, padlock probe molecules are activated prior ligation thus the base at the probe ligation junction is read twice in order to aid successful DNA ligation: first, during structure-specific invader cleavage and then during sequence-specific DNA ligation. We report two distinct iLock probe activation mechanisms and systematically evaluate the assay specificity, including single nucleotide polymorphisms on RNA, mRNA and miRNA. We show significant increase in PBCV-1 ligation fidelity in the iLock probe assay configuration for RNA detection.


Histochemistry as a versatile research toolkit in biological research, not only an applied discipline in pathology.
The impressive progress of histochemistry over the last 50 years has led to setting up specific and sensitive techniques to describe dynamic events, through the detection of specific molecules in the very place where they exist in live cells. The scientific field where histochemistry has most largely been applied is histopathology, with the aim to identify disease-specific molecular markers or to elucidate the etiopathological mechanisms. Numerous authors did however apply histochemistry to a variety of other research fields; their interests range from the microanatomy of animal and plant organisms to the cellular mechanisms of life. This is especially apparent browsing the contents of the histochemical journals where the articles on subjects other than pathology are the majority; these journals still keep a pivotal role in the field of cell and tissue biology, while being a forum for a diverse range of biologists whose scientific interests expand the research horizon of histochemistry to ever novel subjects. Thus, histochemistry can always receive inspiring stimuli toward a continuous methodological refinement.


Trauma-related symptoms in Sri Lankan adult survivors after the tsunami: pretraumatic and peritraumatic factors.
Limited research has addressed factors associated with psychological distress following disasters among non-Western populations. The 2004 tsunami affected 1.7 million people across South Asia and Africa, with considerable variations in trauma-related outcomes. Pretraumatic and peritraumatic conditions associated with trauma-related symptoms in 305 Sri Lankan adult survivors (28% male, aged 18-83 years; mean = 39.9 years; standard deviation = 15.3), clinically assessed 1 month posttsunami, were evaluated retrospectively. Outcome measures were total scores on 11 trauma-related symptoms. Multivariate linear regression analyses tested for associations between pretraumatic and peritraumatic conditions and symptom scores, with peritraumatic conditions adjusted for pretraumatic variables. Pretraumatic conditions of female gender, employment, prior health and social issues, and substance use and peritraumatic conditions of loss of family, witnessing the tsunami, or suffering an injury were associated with trauma-related symptoms. The findings facilitate understanding cultural contexts that define risk factors associated with trauma-related symptoms in Sri Lankans, which are critical for developing culturally appropriate interventions.


Resolution of an intracardiac mass with chemotherapy.
Right atrial intracardiac tumours are uncommonly seen during echocardiography. Our patient presented with primary mediastinal large B-cell lymphoma with intracardiac involvement. The tumour was seen by echocardiography and the extent of the tumour was defined by CT scan of the chest. Following chemotherapy directed to her specific tumour cell type, there was complete resolution of the intracardiac mass.