Regulation of NF-κB-induced inflammatory signaling by lipid peroxidation-derived aldehydes.
Oxidative stress plays a critical role in the pathophysiology of a wide range of diseases including cancer. This view has broadened significantly with the recent discoveries that reactive oxygen species initiated lipid peroxidation leads to the formation of potentially toxic lipid aldehyde species such as 4-hydroxy-trans-2-nonenal (HNE), acrolein, and malondialdehyde which activate various signaling intermediates that regulate cellular activity and dysfunction via a process called redox signaling. The lipid aldehyde species formed during synchronized enzymatic pathways result in the posttranslational modification of proteins and DNA leading to cytotoxicity and genotoxicty. Among the lipid aldehyde species, HNE has been widely accepted as a most toxic and abundant lipid aldehyde generated during lipid peroxidation. HNE and its glutathione conjugates have been shown to regulate redox-sensitive transcription factors such as NF-κB and AP-1 via signaling through various protein kinase cascades. Activation of redox-sensitive transcription factors and their nuclear localization leads to transcriptional induction of several genes responsible for cell survival, differentiation, and death. In this review, we describe the mechanisms by which the lipid aldehydes transduce activation of NF-κB signaling pathways that may help to develop therapeutic strategies for the prevention of a number of inflammatory diseases.


Chemical effects in SERS of pyrazine adsorbed on Au-Pd bimetallic nanoparticles: a theoretical investigation.
Chemical enhancement in surface-enhanced Raman scattering (SERS) of pyrazine adsorbed on Au-Pd nanoclusters is investigated by using density functional theory. Changing Pd content in the bimetallic clusters enables modulation of the direct chemical interactions between the pyrazine and the clusters. The magnitude of chemical enhancement is correlated well with the induced polarizability for the complexes with low Pd content, which fails for the complexes with high Pd content. Furthermore, the dependence of chemical enhancement on cluster size and coupling is also described by the induced polarizability. Additionally, the chemical enhancement in the cluster-molecule-cluster junction is found to account for as much as 10(3), which suggests that a chemical mechanism might be more important than previously believed, in particular for Au-Pd bimetallic nanoparticle aggregates.


The Right Angular Gyrus Combines Perceptual and Response-related Expectancies in Visual Search: TMS-EEG Evidence.
Visual search performance is sensitive to changes in the environment. Attention is sensitive to trial history, in terms of both perception and response. Although the bases of these sensorimotor interactions remain unclear, both behaviorally and neurally, converging evidence from a variety of methods indicates that the right angular gyrus (rANG) may be important. The present study tests whether the rANG plays a causal role in generating these inter-trial effects. Participants performed a compound task for feature singleton targets. We applied rTMS over the rANG (or a control site, or no TMS) during the inter-trial interval and measured effects both on behavior and on neural activity using psychophysics and event-related potential (ERP) recording. rANG TMS during the inter-trial interval improved performance to the upcoming stimuli only when the target-defining dimension and the response-defining feature both repeated across successive trials. rANG TMS also increased the amplitude of the visual N1 component evoked by the upcoming stimuli. These effects did not occur after control TMS. rANG plays a causal role in the formation of combined expectancies binding together stimulus- and response-characteristics of the previous trial to optimize visual search performance. This supports a visuomotor theory of parietal cortex and the dimension weighting account of attention. We suggest current models of inter-trial effects in visual search need to be expanded to include an interactive component representing both perceptual and motoric inter-trial expectancies, affecting the early analysis of stimulus features in the upcoming trial.


The midbrain-hindbrain boundary genetic cascade is activated ectopically in the diencephalon in response to the widespread expression of one of its components, the medaka gene Ol-eng2.
In vertebrates, the engrailed genes are expressed at early neurula stage in a narrow stripe encompassing the midbrain-hindbrain boundary (MHB), a region from which a peculiar structure, the isthmus, is formed. Knock-out experiments in mice demonstrated that these genes are essential for the development of this structure and of its derivatives. In contrast, little is known about the effect of an overexpression of engrailed genes in vertebrate development. Here we report the isolation of Ol-eng2, a medaka fish (Oryzias latipes) engrailed gene. We have monitored the effects of its widespread expression following mRNA injections in 1- and 2-cell medaka and Xenopus embryos. We found that the ectopic expression of Ol-eng2 predominantly results in an altered development of the anterior brain, including an inhibition of optic vesicle formation. No change in the patterns of mesencephalic and telencephalic markers were observed. In contrast, expressions of markers of the diencephalon were strongly repressed in injected embryos. Furthermore, the endogenous Ol-eng2, Pax2, Wnt1 and Fgf8, which are essential components of the MHB genetic cascade, were ectopically expressed in this region. Therefore, we propose that Ol-eng2 induces de novo formation of an isthmus-like structure, which correlates with the development of ectopic midbrain structures, including optic tectum. A competence of the diencephalon to change to a midbrain fate has been demonstrated in isthmic graft experiments. Our data demonstrate that this change can be mimicked by ectopic engrailed expression alone.


Evolutionary genetics of a new pathogenic Escherichia species: Escherichia albertii and related Shigella boydii strains.
A bacterium originally described as Hafnia alvei induces diarrhea in rabbits and causes epithelial damage similar to the attachment and effacement associated with enteropathogenic Escherichia coli. Subsequent studies identified similar H. alvei-like strains that are positive for an intimin gene (eae) probe and, based on DNA relatedness, are classified as a distinct Escherichia species, Escherichia albertii. We determined sequences for multiple housekeeping genes in five E. albertii strains and compared these sequences to those of strains representing the major groups of pathogenic E. coli and Shigella. A comparison of 2,484 codon positions in 14 genes revealed that E. albertii strains differ, on average, at approximately 7.4% of the nucleotide sites from pathogenic E. coli strains and at 15.7% from Salmonella enterica serotype Typhimurium. Interestingly, E. albertii strains were found to be closely related to strains of Shigella boydii serotype 13 (Shigella B13), a distant relative of E. coli representing a divergent lineage in the genus Escherichia. Analysis of homologues of intimin (eae) revealed that the central conserved domains are similar in E. albertii and Shigella B13 and distinct from those of eae variants found in pathogenic E. coli. Sequence analysis of the cytolethal distending toxin gene cluster (cdt) also disclosed three allelic groups corresponding to E. albertii, Shigella B13, and a nontypeable isolate serologically related to S. boydii serotype 7. Based on the synonymous substitution rate, the E. albertii-Shigella B13 lineage is estimated to have split from an E. coli-like ancestor approximately 28 million years ago and formed a distinct evolutionary branch of enteric pathogens that has radiated into groups with distinct virulence properties.


Eye problems in breast cancer patients treated with tamoxifen.
Tamoxifen is an oral anti-estrogen used in the treatment of breast cancer and recently approved to reduce the incidence of breast cancer in high risk women. As a large clinical trial of tamoxifen has reported an increased risk of cataract, we conducted a study of women with breast cancer to evaluate the association of tamoxifen with cataracts and other eye problems. We attempted to recruit previously interviewed patients who were cases in a population-based case-control study of 2653 women with primary breast cancer diagnosed between 1987 and 1996 at ages 55-72 years in Los Angeles County, California, USA. In November 1997, each case was mailed a questionnaire to ascertain self-reported incidence of eye diseases and Amsler grid test scores. Information from 1297 women aged 57-75 years of age was analyzed. Women reporting treatment with tamoxifen were categorized as standard-term users (4-5 years), short-term users (<4 years), or long-term users (6+ years) and compared to non-users. All p-values, relative risks, and confidence intervals for differences in eye problems and grid test results are adjusted for age and stage of disease at diagnosis. Standard-term and long-term users of tamoxifen more frequently reported developing cataracts than non-users (18.2%, 21.4% vs. 14.8%). The relative risk was 1.40 (95% confidence interval 0.94-2.10) for standard-term users and 1.70 (1.11-2.59) for long-term users. Tamoxifen use was unrelated to frequency of glaucoma or macular degeneration or to Amsler grid test results. Our study suggests that five or more years of tamoxifen use increases risk of cataracts. Healthy women considering tamoxifen use to reduce risk of breast cancer should be advised of the possibility of cataract development. Women choosing such therapy should be diligent about receiving regular ocular exams.


Calcyclin-binding site located on the NH2-terminal domain of rabbit CAP-50 (annexin XI): functional expression of CAP-50 in Escherichia coli.
CAP-50 (annexin XI) is a member of annexin family proteins originally identified and characterized as a target protein for calcyclin (H. Tokumitsu et al. (1992) J. Biol. Chem. 267, 8919-8924). In the present work, the calcyclin-binding site of CAP-50 was determined by proteolytic study and by using various deletion mutants expressed in Escherichia coli. The 43-kDa fragment of CAP-50 digested with Staphylococcus aureus V8 protease did not bind to calcyclin in the presence of Ca2+. CAP-50 fusion proteins, including various NH2-terminal deletion mutants were expressed in E. coli using rabbit CAP-50 cDNA and the calcyclin-binding potential was examined using the 125I-calcyclin gel overlay method and coprecipitation with phosphatidylserine-containing vesicles in the presence of Ca2+. All recombinant protein carried the potential for Ca(2+)-dependent phospholipid binding, due to the presence of the COOH-terminal domain (core domain). Calcyclin-binding experiments showed that CAP-50 molecules lacking the NH2-terminal 26 residues retain binding potential for calcyclin; however, deletion of an additional 26 amino acids from the NH2 terminus abolishes specific calcyclin binding. These observations suggest that the calcyclin-binding site is located on the NH2-terminal region of CAP-50, probably adjacent to or within the region from Tyr27 to Leu52.


Strain-dependent differences in LTP and hippocampus-dependent memory in inbred mice.
Many studies have used "reverse" genetics to produce "knock-out" and transgenic mice to explore the roles of various molecules in long-term potentiation (LTP) and spatial memory. The existence of a variety of inbred strains of mice provides an additional way of exploring the genetic bases of learning and memory. We examined behavioral memory and LTP expression in area CA1 of hippocampal slices prepared from four different inbred strains of mice: C57BL/6J, CBA/J, DBA/2J, and 129/SvEms-+(Ter?)/J. We found that LTP induced by four 100-Hz trains of stimulation was robust and long-lasting in C57BL/6J and DBA/2J mice but decayed in CBA/J and 129/SvEms-+(Ter?)/J mice. LTP induced by one 100-Hz train was significantly smaller after 1 hr in the 129/SvEms-+(Ter?)/J mice than in the other three strains. Theta-burst LTP was shorter lasting in CBA/J, DBA/2J, and 129/SvEms-+(Ter?)/J mice than in C57BL/6J mice. We also observed specific memory deficits, among particular mouse strains, in spatial and nonspatial tests of hippocampus-dependent memory. CBA/J mice showed defective learning in the Morris water maze, and both DBA/2J and CBA/J strains displayed deficient long-term memory in contextual and cued fear conditioning tests. Our findings provide strong support for a genetic basis for some forms of synaptic plasticity that are linked to behavioral long-term memory and suggest that genetic background can influence the electrophysiological and behavioral phenotypes observed in genetically modified mice generated for elucidating the molecular bases of learning, memory, and LTP.


Progress in the active immunotherapeutic approach to Alzheimer's disease: clinical investigations into AN1792-associated meningoencephalitis.
In a phase 2a clinical trial of AN1792 (Study 201), a potential immunotherapeutic agent for use in Alzheimer's disease (AD), approximately 6% of the treated AD patients (18/300) developed meningoencephalitis (ME). To elucidate potential immune mechanisms of treatment-induced ME. Peripheral blood mononuclear cells obtained from patients who received AN1792 were stimulated in vitro either with beta-amyloid (Abeta) or various overlapping peptides of Abeta(1-42), followed by quantification of cytokine-secreting cells by enzyme-linked immunosorbent spot assay. A significant difference in the quality of the T-cell responses between patients in Study 201 and those in earlier studies of AN1792 was noted. T-cell responses specific to the carboxy terminus of Abeta elicited from patients' peripheral blood mononuclear cells in an earlier multiple dose study (Study 102) were Th2 biased, while those from Study 201 were biased toward a proinflammatory Th1 response. Antibody responses in both studies were quantitatively and qualitatively similar (as determined by epitope mapping). The addition of polysorbate 80 to the formulation used in Study 201 is the most likely explanation for the difference in the T-cell response. ME following injection of AN1792 may be related to immune response differences driven by a formulation change. To address this, a novel peptide-carrier protein conjugate using an amino-terminal fragment of Abeta (ACC-001) has been developed to avoid potentially harmful T-cell responses, while maintaining a similar antibody response to that of AN1792. Immunotherapeutic trials using this treatment approach in AD patients are in progress.


A predictive model for ADL dependence in community-living older adults based on a reduced set of cognitive status items.
To develop and validate a simple tool, based on a reduced set of Mini-Mental State Examination (MMSE) items, that can be used to predict the onset of ADL dependence, and to compare the predictive accuracy of this new tool with that of the MMSE. Two prospective, population-based cohort studies, in tandem. The predictive model developed in the initial cohort was subsequently validated in a separate cohort. General community in New Haven, Connecticut. For the development cohort, 775 community-living persons, 72 years of age and older, who were independent at baseline in their ADI, function. For the validation cohort, 1038 comparable subjects. All subjects underwent a baseline interview and cognitive assessment in their homes by a trained research nurse using standard instrument. Self-reported ADLs were ascertained at 1 year and 3 years for the development cohort and at 1 year and 21/2 years for the validation cohort. ADL dependence developed in 221 (28.5%) subjects in the development cohort. Although the rate of ADL dependence increased within each MMSE domain as the number of incorrect items increased, only orientation and short-term memory remained significantly associated with ADL dependence in multivariable analysis. A predictive model, based on the presence of impairments in these two domains, was developed that stratified subjects into three risk groups. Rates of ADL dependence were 22% (neither domain impaired), 44% (one domain impaired), and 68% (both domains impaired) (P < .001). The corresponding rates in the validation cohort, in which 191 (18.4%) subjects developed ADL dependence, were 15%, 26%, and 45% (P < .001). The area under the ROC curves for the MMSE and the reduced item strategy were nearly identical at 0.63 and 0.62, respectively. A simple and valid six-item strategy, based on the presence of impairments in orientation and short-term memory, predicts the onset of ADL dependence as effectively as does the 30-item MMSE. This new tool may be useful as part of a more comprehensive assessment when determining an older person's risk for developing ADL dependence.


A monoclonal antibody directed against a Candida albicans cell wall mannoprotein exerts three anti-C. albicans activities.
Antibodies are believed to play a role in the protection against Candida albicans infections by a number of mechanisms, including the inhibition of adhesion or germ tube formation, opsonization, neutralization of virulence-related enzymes, and direct candidacidal activity. Although some of these biological activities have been demonstrated individually in monoclonal antibodies (MAbs), it is not clear if all these anti-C. albicans activities can be displayed by a single antibody. In this report, we characterized a monoclonal antibody raised against the main target of salivary secretory immunoglobulin A in the cell wall of C. albicans, which exerts three anti-C. albicans activities: (i) inhibition of adherence to HEp-2 cells, (ii) inhibition of germination, and (iii) direct candidacidal activity. MAb C7 reacted with a proteinic epitope from a mannoprotein with a molecular mass of >200 kDa predominantly expressed on the C. albicans germ tube cell wall surface as well as with a number of antigens from Candida lusitaniae, Cryptococcus neoformans, Aspergillus fumigatus, and Scedosporium prolificans. MAb C7 caused a 31.1% inhibition in the adhesion of C. albicans to HEp-2 monolayers and a 55.3% inhibition in the adhesion of C. albicans to buccal epithelial cells, produced a 38.5% decrease in the filamentation of C. albicans, and exhibited a potent fungicidal effect against C. albicans, C. lusitaniae, Cryptococcus neoformans, A. fumigatus, and S. prolificans, showing reductions in fungal growth ranging from 34.2 to 88.7%. The fungicidal activity showed by MAb C7 seems to be related to that reported by antibodies mimicking the activity of a killer toxin produced by the yeast Pichia anomala, since one of these MAbs also reacted with the C. albicans mannoprotein with a molecular mass of >200 kDa. Results presented in this study support the concept of a family of microbicidal antibodies that could be useful in the treatment of a wide range of microbial infections when used alone or in combination with current antimicrobial agents.


Protein sequences conserved in prokaryotic aminoacyl-tRNA synthetases are important for the activity of the processivity factor of human mitochondrial DNA polymerase.
Previous studies have shown that the small subunit of Xenopus DNA polymerase gamma (pol gammaB) acts as a processivity factor to stimulate the 140 kDa catalytic subunit of human DNA polymerase gamma. A putative human pol gammaB initially identified by analysis of DNA sequence had not been shown to be functional, and appeared to be an incomplete clone. In this paper, we report the cloning of full-length human and mouse pol gammaB. Both human and mouse pol gammaB proteins were expressed in their mature forms, without their apparent mitochondrial localization signals, and shown to stimulate processivity of the recombinant catalytic subunit of human pol gammaA. Deletion analysis of human pol gammaB indicated that blocks of sequence conserved with prokaryotic class II aminoacyl-tRNA synthetases are necessary for activity and inter-action with human pol gammaA. Purification of DNA pol gamma from HeLa cells indicated that both proteins are associated in vivo.


Immediate free TRAM reconstruction in lumpectomy and radiation failure patients.
Local recurrence after lumpectomy and radiation therapy indicates failed breast conservation surgery. These patients often proceed to mastectomy and are candidates for autogenous breast reconstruction. Free transverse rectus abdominus muscle (TRAM) reconstruction in these patients is complicated by repeated axillary dissection and the use of irradiated tissue. Complication rates for pedicled TRAMs have been reported at 33 percent when used in irradiated tissue beds. We report our results using the free TRAM for breast reconstruction after lumpectomy and radiation failure. All patients within this study developed a local recurrence after lumpectomy and radiation therapy. All patients had undergone axillary dissection for staging at the time of their lumpectomy. Patient records were reviewed for patient age, total radiation dose, associated risk factors for TRAM failure, operative time, donor vessels used for anastomosis, status of the native thoracodorsal vessels at the time of surgery, and postoperative complications. Over a 7-year period, 16 TRAM patients had undergone previous breast conservation surgery. Of these 16 patients, 14 underwent reconstruction with a planned free TRAM after simple mastectomy. Average operating room time was 7 hours. There were no partial or total flap losses. Complications were seen in 14 percent of the overall group. Overall, we found that the free TRAM provided an excellent aesthetic result with a lower complication rate than previously reported for pedicled TRAM flaps in irradiated beds. The thoracodorsal vessels provided an adequate donor vessel in 93 percent of the cases. The free TRAM provides a superior alternative in immediate reconstruction in patients who have failed breast conservative surgery.


Molecular determinants of response to matuzumab in combination with paclitaxel for patients with advanced non-small cell lung cancer.
Antibodies targeting epidermal growth factor receptor (EGFR) have proven to be effective in patients with non-small cell lung cancer (NSCLC) that express EGFR. We recently published a phase I study of weekly matuzumab plus paclitaxel. This therapy was well tolerated and showed clinical responses in the majority of patients. Although matuzumab displays potent antitumor activity in some patients, not all patients respond well to treatment. Whether dysregulation of EGFR-mediated pathways precludes or sensitizes cells to paclitaxel is unknown. We sought to determine molecular predictive factors for therapy response in a phase I/II study patient cohort treated with matuzumab+/-paclitaxel. Twenty-three cases [including one complete response (CR), three partial responses (PR), 10 stable diseases (SD)] were screened using immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), PCR/sequencing and denaturing wave high performance liquid chromatography (D-HPLC) for expression, amplification, and mutation status of EGFR and downstream signaling pathways. All patients with PR or CR displayed an either high overall or single-cell EGFR expression in the majority of cells. In addition, all of the moderate responders, who achieved SD after at least two cycles of therapy, showed diffuse EGFR expression rates and/or strong single-cell EGFR expression. In contrast, 44% of the nonresponders showed low overall or single-cell EGFR expression levels. No low-expressing EGFR cases were present within the responder group. In addition, among patients with a gain-of-function mutation in KRAS primary therapy failure and/or short responses to therapy were observed. Our data suggest that EGFR expression and KRAS mutation status is predictive for clinical response to matuzumab +/- paclitaxel in patients with advanced NSCLC.


Dispersal and group formation dynamics in a rare and endangered temperate forest bat (Nyctalus lasiopterus, Chiroptera: Vespertilionidae).
For elusive mammals like bats, colonization of new areas and colony formation are poorly understood, as is their relationship with the genetic structure of populations. Understanding dispersal and group formation behaviors is critical not only for a better comprehension of mammalian social dynamics, but also for guiding conservation efforts of rare and endangered species. Using nuclear and mitochondrial markers, we studied patterns of genetic diversity and differentiation among and within breeding colonies of giant noctule bats (Nyctalus lasiopterus), their relation to a new colony still in formation, and the impact of this ongoing process on the regionwide genetic makeup. Nuclear differentiation among colonies was relatively low and mostly nonsignificant. Mitochondrial variation followed this pattern, contrasting with findings for other temperate bat species. Our results suggest that this may indicate a recent population expansion. On average, female giant noctules were not more closely related to other colony members than to foreign individuals. This was also true for members of the newly forming colony and those of another, older group sampled shortly after its formation, suggesting that contrary to findings for other temperate bats, giant noctule colonies are not founded by relatives. However, mother-daughter pairs were found in the same populations more often than expected under random dispersal. Given this indication of philopatry, the lack of mitochondrial differentiation among most colonies in the region is probably due to the combination of a recent population expansion and group formation events.


Cost-effectiveness of 12-month treatment with ticagrelor compared with clopidogrel in the management of acute coronary syndromes.
The PLATO (Platelet Inhibition and Patient Outcomes) randomized trial (NCT00391872) in patients with acute coronary syndromes (ACS) reported that ticagrelor (in addition to aspirin) reduced the rate of the composite end point of myocardial infarction (MI), stroke, or cardiovascular death compared with clopidogrel (in addition to aspirin) by 16% over 12 months (P < 0.001). No significant difference in the incidence of major bleeding was noted, but ticagrelor was associated with a higher rate of major bleeding not related to coronary artery bypass grafting. By extrapolating the key findings of PLATO, we sought to assess the cost-effectiveness of ticagrelor compared with clopidogrel in the management of ACS in a contemporary Australian setting. A Markov model with 4 health states (free from further ACS events, MI, stroke, and death) was developed to simulate the long-term costs and outcomes associated with ACS. Event risks were based on data derived directly from PLATO, and costs and utilities were drawn from published sources. A 10-year time horizon was simulated, and future costs and benefits were discounted at a 5% annual rate. However, treatment with ticagrelor and clopidogrel was only assumed for the first 12 months, with no benefits applied beyond drug cessation. Sensitivity analyses were undertaken based on variations to key data inputs. All costs for resource use applied in the analysis were based on published Australian prices (in 2010/2011 dollars [A$]). Over 10 years, the estimated quality-adjusted life-years lived per-patient were 5.74 and 5.68 for ticagrelor and clopidogrel, respectively. Net costs were A$19,132 for ticagrelor and A$18,428 for clopidogrel. These equated to an incremental cost-effectiveness ratio of A$9031 per quality-adjusted life-year gained for ticagrelor compared with clopidogrel. Sensitivity analyses indicated the result to be robust. When assessed from the perspective of the Australian health care system, ticagrelor is likely to be cost-effective compared with clopidogrel in preventing downstream morbidity and mortality associated with ACS.


Fine analysis of spontaneous MAGE-C1/CT7-specific immunity in melanoma patients.
Cancer/testis (CT) antigens represent prime candidates for immunotherapy in cancer patients, because their expression is restricted to cancer cells and germ cells of the testis. MAGE-C1/CT7 is a CT antigen that is highly expressed in several types of cancers. Spontaneous occurrence of CT7-specific antibodies was previously detected by SEREX screen in a melanoma patient. However, naturally occurring CT7-specific T-cell responses have thus far not been detected. Peripheral blood mononuclear cells (PBMCs) from 26 metastatic melanoma patients expressing CT7 in their tumor lesions (CT7(+)) were analyzed for CT7-specific T-cell responses using overlapping peptides. CT7-specific CD4(+) T-cell responses were detected in three patients (11.5%). These CT7-specific CD4(+) T-cell responses were detectable in melanoma patients' PBMCs exclusively from preexisting CD45RA(-) memory CD4(+) T-cell pool. Additional CT7-specific memory CD4(+) T-cell responses were detected in CT7(+) melanoma patients after depletion of CD4(+)CD25high Treg cells showing that Treg cells impact on CT7-specific CD4(+) T cells in melanoma patients. CT7-specific CD4(+) T-cell clones were generated and used to define minimal epitopes, restriction elements, and confirm the recognition of naturally processed antigen. Surprisingly, these clones were able to secrete perforin and exert cytotoxicity. This study shows that CT7 can induce specific cellular immunity in melanoma patients. Based on these findings, CT7 will be further explored as a potential vaccine for melanoma immunotherapy.


Differences in anti-apoptotic and multidrug resistance phenotypes in elderly and young acute myeloid leukemia patients are related to the maturation of blast cells.
Elderly patients with acute myeloid leukemia (AML) have a less favorable outcome, which has been related, among other factors, to multidrug resistance (MDR) phenotypes. Freshly obtained erythrocyte-lysed bone marrow samples from 150 elderly patients (> 65 years) with de novo AML and 30 younger AML patients were analyzed using a 4-color immunofluorescence technique for quantitative expression of proteins associated with apoptosis (bcl-2, bax, APO2.7) and MDR (P-gp, MRP, LRP) in 3 blast cell subpopulations, defined according to their maturation stage. Although a homogeneous CD34+ blast cell population was more frequent in the elderly patients, (25% vs 7%, p=0.02), no statistically significant differences were detected between the two age groups in the expression of either apoptosis- or MDR-associated proteins, except for slightly higher quantities of LRP protein in the more immature CD34+ blast cell subset in the elderly AML cases (p=0.04). Interestingly, when different blast cell populations were compared, immature (CD34+) blast cells were characterized by higher levels of bcl-2 in both age groups and lower levels of APO2.7 in the elderly group. In addition, higher P-gp levels were found in CD34+ blast cells than in CD34-- ones in elderly AML patients. Reactivity for LRP was low in both elderly and younger patients. In summary, our results suggest that the higher resistance to chemotherapy observed in elderly AML patients could be related to a higher incidence of cases with a CD34+ homogeneous blast cell population, since these blast cells frequently display a more pronounced anti-apoptotic and MDR1 phenotype.


Investigating the Turing conditions for diffusion-driven instability in the presence of a binding immobile substrate.
Turing's diffusion-driven instability for the standard two species reaction-diffusion system is only achievable under well-known and rather restrictive conditions on both the diffusion rates and the kinetic parameters, which necessitates the pairing of a self-activator with a self-inhibitor. In this study we generalize the standard two-species model by considering the case where the reactants can bind to an immobile substrate, for instance extra-cellular matrix, and investigate the influence of this dynamics on Turing's diffusion-driven instability. Such systems have been previously studied on the grounds that binding of the self-activator to a substrate may effectively reduce its diffusion rate and thus induce a Turing instability for species with equal diffusion coefficients, as originally demonstrated by Lengyel and Epstein (1992) under the assumption that the bound state dynamics occurs on a fast timescale. We, however, analyse the full system without any separation of timescales and demonstrate that the full system also allows a relaxation of the standard constraints on the reaction kinetics for the Turing instability, increasing the type of interactions that could give rise to spatial patterning. In particular, we show that two self-activators can undertake a diffusively driven instability in the presence of a binding immobile substrate, highlighting that the interactions required of a putative biological Turing instability need not be associated with a self-activator-self-inhibitor morphogen pair.


AGA White Paper: Training and Implementation of Endoscopic Image Enhancement Technologies.
Endoscopic image-enhancement technologies provide opportunities to visualize normal and abnormal tissues within the gastrointestinal (GI) tract in a manner that complements conventional white light endoscopic imaging. The additional information that is obtained enables the endoscopist to better identify, delineate, and characterize lesions and can facilitate targeted biopsies or, in some cases, eliminate the need to send samples for histologic analysis. Some of these technologies have been available for more than a decade, but despite this fact, there is limited use of these technologies by endoscopists. Lack of formalized training in their use and a scarcity of guidelines on implementation of these technologies into clinical practice are contributing factors. In November 2014, the American Gastroenterological Association's Center for GI Innovation and Technology conducted a 2-day workshop to discuss endoscopic image-enhancement technologies. This article represents the third of 3 separate documents generated from the workshop and discusses the published literature pertaining to training and outlines a proposed framework for the implementation of endoscopic image-enhancement technologies in clinical practice. There was general agreement among participants in the workshop on several key considerations. Training and competency assessment for endoscopic image-enhancement technologies should incorporate competency-based education paradigms. To facilitate successful training, multiple different educational models that can cater to variations in learning styles need to be developed, including classroom-style and self-directed programs, in-person and web-based options, image and video atlases, and endoscopic simulator programs. To ensure safe and appropriate use of these technologies over time, refresher courses, skill maintenance programs, and options for competency reassessment should be established. Participants also generally agreed that although early adopters of novel endoscopic image-enhancement modalities can successfully implement these technologies by pursuing training and ensuring self-competency, widespread implementation is likely to require support from GI societies and buy-in from other key stakeholders including payors/purchasers and patients. Continued work by manufacturers and the GI societies in providing training programs and patient education, working with payors and purchasers, and creating environments and policies that motivate endoscopists to adopt new practices is essential in creating widespread implementation.


Efficacy of virtual reality in pedestrian safety research.
Advances in virtual reality technology present new opportunities for human factors research in areas that are dangerous, difficult, or expensive to study in the real world. The authors developed a new pedestrian simulator using the HTC Vive head mounted display and Unity software. Pedestrian head position and orientation were tracked as participants attempted to safely cross a virtual signalized intersection (5.5 m). In 10% of 60 trials, a vehicle violated the traffic signal and in 10.84% of these trials, a collision between the vehicle and the pedestrian was observed. Approximately 11% of the participants experienced simulator sickness and withdrew from the study. Objective measures, including the average walking speed, indicate that participant behavior in VR matches published real world norms. Subjective responses indicate that the virtual environment was realistic and engaging. Overall, the study results confirm the effectiveness of the new virtual reality technology for research on full motion tasks.


Function and Distribution of a Lantipeptide in Strawberry Fusarium Wilt Disease-Suppressive Soils.
Streptomyces griseus S4-7 is representative of strains responsible for the specific soil suppressiveness of Fusarium wilt of strawberry caused by Fusarium oxysporum f. sp. fragariae. Members of the genus Streptomyces secrete diverse secondary metabolites including lantipeptides, heat-stable lanthionine-containing compounds that can exhibit antibiotic activity. In this study, a class II lantipeptide provisionally named grisin, of previously unknown biological function, was shown to inhibit F. oxysporum. The inhibitory activity of grisin distinguishes it from other class II lantipeptides from Streptomyces spp. Results of quantitative reverse transcription-polymerase chain reaction with lanM-specific primers showed that the density of grisin-producing Streptomyces spp. in the rhizosphere of strawberry was positively correlated with the number of years of monoculture and a minimum of seven years was required for development of specific soil suppressiveness to Fusarium wilt disease. We suggest that lanM can be used as a diagnostic marker of whether a soil is conducive or suppressive to the disease.


Influence of carrier transport on Raman amplification in silicon waveguides.
This paper analyze for the first time the impact of the charge carrier transport on the continuously pumped stimulated Raman amplification in silicon waveguides. A novel analytical model is developed using which the coupled differential equations of the pump and the probe optical signals and those of carrier transport are solved concurrently. The simulation and analysis suggest that the neglect of the carrier transport phenomenon, reported in the previously published works, is approximately justified only if the effective carrier lifetime is comparable to the carrier transit time, otherwise it can result in substantial overestimation of the free carrier density at the optical mode center.


Mapping stain distribution in pathology slides using whole slide imaging.
Whole slide imaging (WSI) offers a novel approach to digitize and review pathology slides, but the voluminous data generated by this technology demand new computational methods for image analysis. In this study, we report a method that recognizes stains in WSI data and uses kernel density estimator to calculate the stain density across the digitized pathology slides. The validation study was conducted using a rat model of acute cardiac allograft rejection and another rat model of heart ischemia/reperfusion injury. Immunohistochemistry (IHC) was conducted to label ED1(+) macrophages in the tissue sections and the stained slides were digitized by a whole slide scanner. The whole slide images were tessellated to enable parallel processing. Pixel-wise stain classification was conducted to classify the IHC stains from those of the background and the density distribution of the identified IHC stains was then calculated by the kernel density estimator. The regression analysis showed a correlation coefficient of 0.8961 between the number of IHC stains counted by our stain recognition algorithm and that by the manual counting, suggesting that our stain recognition algorithm was in good agreement with the manual counting. The density distribution of the IHC stains showed a consistent pattern with those of the cellular magnetic resonance (MR) images that detected macrophages labeled by ultrasmall superparamagnetic iron-oxide or micron-sized iron-oxide particles. Our method provides a new imaging modality to facilitate clinical diagnosis. It also provides a way to validate/correlate cellular MRI data used for tracking immune-cell infiltration in cardiac transplant rejection and cardiac ischemic injury.


A single polymorphic residue within the peptide-binding cleft of MHC class I molecules determines spectrum of tapasin dependence.
Different HLA class I alleles display a distinctive dependence on tapasin for surface expression and Ag presentation. In this study, we show that the tapasin dependence of HLA class I alleles correlates to the nature of the amino acid residues present at the naturally polymorphic position 114. The tapasin dependence of HLA class I alleles bearing different residues at position 114 decreases in the order of acidity, with high tapasin dependence for acidic amino acids (aspartic acid and glutamic acid), moderate dependence for neutral amino acids (asparagine and glutamine), and low dependence for basic amino acids (histidine and arginine). A glutamic acid to histidine substitution at position 114 allows the otherwise tapasin-dependent HLA-B4402 alleles to load high-affinity peptides independently of tapasin and to have surface expression levels comparable to the levels seen in the presence of tapasin. The opposite substitution, histidine to glutamic acid at position 114, is sufficient to change the HLA-B2705 allele from the tapasin-independent to the tapasin-dependent phenotype. Furthermore, analysis of point mutants at position 114 reveals that tapasin plays a principal role in transforming the peptide-binding groove into a high-affinity, peptide-receptive conformation. The natural polymorphisms in HLA class I H chains that selectively affect tapasin-dependent peptide loading provide insights into the functional interaction of tapasin with MHC class I molecules.


Quantitative X-ray wavefront measurements of Fresnel zone plate and K-B mirrors using phase retrieval.
A scanning coherent diffraction imaging method was used to reconstruct the X-ray wavefronts produced by a Fresnel zone plate (FZP) and by Kirkpatrick-Baez (KB) focusing mirrors. The ptychographical measurement was conducted repeatedly by placing a lithographed test sample at different defocused planes. The wavefronts, recovered by phase-retrieval at well-separated planes, show good consistency with numerical propagation results, which provides a self-verification. The validity of the obtained FZP wavefront was further confirmed with theoretical predictions.


Bootstrap percolation on spatial networks.
Bootstrap percolation is a general representation of some networked activation process, which has found applications in explaining many important social phenomena, such as the propagation of information. Inspired by some recent findings on spatial structure of online social networks, here we study bootstrap percolation on undirected spatial networks, with the probability density function of long-range links' lengths being a power law with tunable exponent. Setting the size of the giant active component as the order parameter, we find a parameter-dependent critical value for the power-law exponent, above which there is a double phase transition, mixed of a second-order phase transition and a hybrid phase transition with two varying critical points, otherwise there is only a second-order phase transition. We further find a parameter-independent critical value around -1, about which the two critical points for the double phase transition are almost constant. To our surprise, this critical value -1 is just equal or very close to the values of many real online social networks, including LiveJournal, HP Labs email network, Belgian mobile phone network, etc. This work helps us in better understanding the self-organization of spatial structure of online social networks, in terms of the effective function for information spreading.


Active control of a 30 m ring interferometric telescope primary mirror.
The active control of a primary mirror for a ring aperture segmented telescope is different from that of a full aperture segmented telescope. Two active maintenance proposals for the ring telescope designed from the segmented patterns are outlined. We present a preliminary calculation of noise propagation and analysis of primary mirror mode characteristics for each proposal. The modulation transfer functions (MTFs) of the primary mirror corresponding to each maintenance method at several typical working wavelengths are also simulated.


Smoking dependence in 18 European countries: Hard to maintain the hardening hypothesis.
When the prevalence of smoking decreases in a population, there is a hypothesis-the so-called "hardening hypothesis"-that the remaining smokers form a subgroup of "hardcore smokers." Our aims were to test the hardening hypothesis and to analyze the determinants of high dependence taking into account both individual and country-level characteristics. Within the Pricing Policies and Control of Tobacco in Europe (PPACTE) project, we conducted a face-to-face survey on smoking between January and July 2010 in 18 European countries, including 2882 male and 2254 female smokers with complete information on smoking dependence. The Heaviness of Smoking Index (HSI) was used as a measure of tobacco dependence. We correlated smoking prevalence and dependence using the country as unit of analysis. Moreover, we fitted multilevel logistic regression models. Country-specific prevalence of smoking was positively, although not significantly, correlated with the proportion of highly tobacco-dependent smokers (overall rsp=0.203, p=0.419), both in men (rsp=0.235, p=0.347) and women (rsp=0.455, p=0.058). Using individual-level analysis, high dependence was positively related to age, and, although not significantly, to smoking prevalence, and inversely related to level of education. The lack of a smoking ban at home was positively related to smoking dependence. Using both ecological and individual-level analyses, the relations between smoking prevalence and HSI were not significant, but in the opposite direction as compared to that assumed by the "hardening hypothesis." Therefore, our data provide empirical evidence against this theory, thus supporting the feasibility of an endgame strategy.


Activation of transcription factor Nrf2 signalling by the sphingosine kinase inhibitor SKI-II is mediated by the formation of Keap1 dimers.
Anti-oxidant capacity is crucial defence against environmental or endogenous oxidative stress. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a redox-sensitive transcription factor that plays a key defensive role against oxidative and cytotoxic stress and cellular senescence. However, Nrf2 signalling is impaired in several aging-related diseases, such as chronic pulmonary obstructive disease (COPD), cancer, and neurodegenerative diseases. Thus, novel therapeutics that enhance Nrf2 signalling are an attractive approach to treat these diseases. Nrf2 was stabilized by SKI-II (2-(p-hydroxyanilino)-4-(p-chlorophenyl) thiazole), which is a known sphingosine kinase inhibitor, in human bronchial epithelial cell line, BEAS2B, and in primary human bronchial epithelial cells, leading to enhancement of anti-oxidant proteins, such as HO-1, NQO1 and GCLM. The activation of Nrf2 was achieved by the generation of inactive dimerized form of Keap1, a negative regulator of Nrf2 expression, which was independent of sphingosine kinase inhibition. Using mice that were exposed to cigarette smoke, SKI-II induced Nrf2 expression together with HO-1 in their lungs. In addition, SKI-II reduced cigarette smoke mediated oxidative stress, macrophages and neutrophil infiltration and markers of inflammation in mice. SKI-II appears to be a novel activator of Nrf2 signalling via the inactivation of Keap1.


Iodine-catalyzed sulfenylation of pyrazolones using dimethyl sulfoxide as an oxidant.
An iodine catalyzed sulfenylation of pyrazolones with a diverse range of heterocyclic thiols, heterocyclic thiones and disulfides has been described using dimethyl sulfoxide as an oxidant, which is an inexpensive, readily available and green oxidant. The present methodology exhibits a wide range of substrate scope and targeted products were obtained in good to excellent yields under metal-free conditions in a short duration. This methodology provides a simple process for the formation of C-S bonds through the thioetherification of pyrazolones.


Fetal growth and insulin resistance in adult life: role of skeletal muscle morphology.
1. Thinness at birth is associated with insulin resistance in adult life and an apparent delay in activation of glycolysis/glycogenolysis in exercising skeletal muscle. As developmental abnormalities of skeletal muscle histology or metabolism may explain this association we examined muscle histology, biochemistry and blood flow in a group of 27 adult women whose birth details were known. 2. Subjects were examined by near-infrared spectroscopy to determine forearm muscle oxygen supply, and by muscle biopsy and forearm plethysmography. Those with a ponderal index at birth < 23 kg/m3 were insulin resistant (assessed by the short insulin-tolerance test-mean rate constants for glucose disappearance = 4.14 compared with 4.83%/min, P = 0.045) and had significantly more rapid muscle reoxygenation than the remainder of the subjects (13 compared with 22 s, P = 0.004). 3. Thinness at birth did not influence muscle capillary density, muscle glycogen content, glycogen synthase activity, citrate synthase activity or resting forearm blood flow. 4. Insulin resistance seen after fetal malnutrition was not associated with abnormal muscle histology, resting muscle blood flow, mitochondrial volume or glycogen content. 5. The increase in muscle reoxygenation rate in adult subjects who were thin at birth could occur to promote oxidative ATP synthesis in compensation for the delay in activation of glycolysis/glycogenolysis. It suggests altered regulation rather than structure of the muscle microcirculation. These changes appear to antedate the structural and biochemical changes seen in muscle from patients with established diabetes.


Potential Pasture Nitrogen Concentrations and Uptake from Autumn or Spring Applied Cow Urine and DCD under Field Conditions.
Nitrogen (N) cycling and losses in grazed grassland are strongly driven by urine N deposition by grazing ruminants. The objective of this study was to quantify pasture N concentrations, yield and N uptake following autumn and spring deposition of cow urine and the effects of fine particle suspension (FPS) dicyandiamide (DCD). A field plot study was conducted on the Lincoln University dairy farm, Canterbury, New Zealand from May 2003 to May 2005. FPS DCD was applied to grazed pasture plots at 10 kg·ha(-1) in autumn and spring in addition to applied cow urine at a N loading rate of 1000 kg·N·ha(-1), with non-urine control plots. Pasture N ranged between 1.9 and 4.8% with higher concentrations from urine. Results indicated that urine consistently increased N concentrations for around 220 days post deposition (mid December/early summer) at which point concentrations dropped to background levels. In urine patches, pasture yield and annual N uptake were dramatically increased on average by 51% for autumn and 28% for spring applied urine, in both years, when DCD was applied. This field experiment provides strong evidence that annual pasture N uptake is more strongly influenced by high urine N deposition than pasture N concentrations. FPS DCD has the potential to result in very high N uptake in urine patches, even when they are autumn deposited.


The effects of some soil characteristics on radon emanation and diffusion.
The effects of moisture content, grain size, temperature, major elemental composition, and the pH of soils on the radon emanation and diffusion coefficients were evaluated in this study. The emanation and diffusion coefficients are strongly influenced by moisture content and grain size. The radon emanation coefficient increased and the diffusion coefficient decreased with decreasing particle size. However, for soils with large particle sizes, the radon emanation and diffusion coefficient remain almost unchanged with variation in grain size. Comparing five different sized soil particles, the emanation coefficient increased and the diffusion coefficient decreased with moisture content. The radon emanation coefficient reached a constant value with different moisture contents depending on the range of grain sizes. The saturation emanation coefficient for less than 0.1, 0.1-0.2, 0.2-0.3, 0.3-0.5, and more than 0.5 mm sized soil grain ranges are 0.47, 0.42, 0.35, 0.26 and 0.23, respectively, with saturation moisture contents of 16%, 14%, 10%, 6% and 4%, respectively. A drastic increase in radon emanation is found at smaller grain sizes with increasing moisture content. Based on the content of major elements and pH of the soils, the multiple regression indicates that the radon emanation coefficient appears to be significantly dependent on iron content and pH. Effective diffusion coefficient values calculated in our study agree with the results calculated by a previous model. Experimental values show that the temperature dependence of the radon diffusion coefficient follows Arrhenius behavior.


Acoustic and perceptual analysis of the sibilant sound /s/ before and after orthognathic surgery.
Orthognathic surgery may have a positive or negative effect on speech. Perceptual evaluation of presurgical and postsurgical articulation is difficult because speech errors, when they occur, are usually fricative distortions, which may be difficult to document reliably. In this study, acoustic analysis was used to supplement perceptual judgment of presurgical and postsurgical productions of /s/. The study population consisted of 9 Cantonese speakers undergoing osteotomy for Class III skeletal deformity and 9 age- and gender-matched adults with normal occlusion and speech. The speech sample consisted of 6 words with the initial sibilant sound /s/. Perceptual analysis included narrow phonetic transcription and classification of error types. Acoustic analysis included measurement of first and second spectral peaks, fricative duration, noise bandwidth, and noise-to-vowel decibel ratio. The results of the perceptual analysis showed a decrease in articulatory errors for the group after surgery, although 5 patients had no perceptual errors before surgery. Acoustic analysis showed significant differences between the experimental and control groups before surgery for 2 variables (spectral peak I and bandwidth). Three months after surgery there were no significant differences between the control group and the experimental group, except for bandwidth. Twelve months after surgery, there were significant differences between the 2 groups in noise bandwidth and spectral peak II. The results suggest a possible relapse at 1 year after surgery, based on spectral peak values. Osteotomy appears to result in a positive change in articulation for most patients, but speech outcome after osteotomy must be evaluated both 1 year and shortly after surgery.


Ferroptosis: process and function.
Ferroptosis is a recently recognized form of regulated cell death. It is characterized morphologically by the presence of smaller than normal mitochondria with condensed mitochondrial membrane densities, reduction or vanishing of mitochondria crista, and outer mitochondrial membrane rupture. It can be induced by experimental compounds (e.g., erastin, Ras-selective lethal small molecule 3, and buthionine sulfoximine) or clinical drugs (e.g., sulfasalazine, sorafenib, and artesunate) in cancer cells and certain normal cells (e.g., kidney tubule cells, neurons, fibroblasts, and T cells). Activation of mitochondrial voltage-dependent anion channels and mitogen-activated protein kinases, upregulation of endoplasmic reticulum stress, and inhibition of cystine/glutamate antiporter is involved in the induction of ferroptosis. This process is characterized by the accumulation of lipid peroxidation products and lethal reactive oxygen species (ROS) derived from iron metabolism and can be pharmacologically inhibited by iron chelators (e.g., deferoxamine and desferrioxamine mesylate) and lipid peroxidation inhibitors (e.g., ferrostatin, liproxstatin, and zileuton). Glutathione peroxidase 4, heat shock protein beta-1, and nuclear factor erythroid 2-related factor 2 function as negative regulators of ferroptosis by limiting ROS production and reducing cellular iron uptake, respectively. In contrast, NADPH oxidase and p53 (especially acetylation-defective mutant p53) act as positive regulators of ferroptosis by promotion of ROS production and inhibition of expression of SLC7A11 (a specific light-chain subunit of the cystine/glutamate antiporter), respectively. Misregulated ferroptosis has been implicated in multiple physiological and pathological processes, including cancer cell death, neurotoxicity, neurodegenerative diseases, acute renal failure, drug-induced hepatotoxicity, hepatic and heart ischemia/reperfusion injury, and T-cell immunity. In this review, we summarize the regulation mechanisms and signaling pathways of ferroptosis and discuss the role of ferroptosis in disease.


A comparative theoretical study of dipeptide solvation in water.
Molecular dynamics studies have been performed on the zwitterionic form of the dipeptide glycine-alanine in water, with focus on the solvation and electrostatic properties using a range of theoretical methods, from purely classical force fields, through mixed quantum mechanical/molecular mechanical simulations, to fully quantum mechanical Car-Parrinello calculations. The results of these studies show that the solvation pattern is similar for all methods used for most atoms in the dipeptide, but can differ substantially for some groups; namely the carboxy and aminoterminii, and the backbone amid NH group. This might have implications in other theoretical studies of peptides and proteins with charged -NH(3) (+) and -CO(2) (-) side chains solvated in water. Hybrid quantum mechanical/molecular mechanical simulations successfully reproduce the solvation patterns from the fully quantum mechanical simulations (PACS numbers: 87.14.Ee, 87.15.Aa, 87.15.He, 71.15.Pd).


A three-dimensional computer model analysis of three hypothetical biofilm detachment mechanisms.
Three hypothetical mechanisms of detachment were incorporated into a three-dimensional computer model of biofilm development. The model integrated processes of substrate utilization, substrate diffusion, growth, cell advection, and detachment in a cellular automata framework. The purpose of this investigation was to characterize each of the mechanisms with respect to four criteria: the resulting biofilm structure, the existence of a steady state, the propensity for sloughing events, and the dynamics during starvation. The three detachment mechanisms analyzed represented various physical and biological influences hypothesized to affect biofilm detachment. The first invoked the concept of fluid shear removing biomass that protrudes far above the surface and is therefore subjected to relatively large drag forces. The second pathway linked detachment to changes in the local availability of a nutrient. The third pathway simulated an erosive process in which individual cells are lost from the surface of a biofilm cell cluster. The detachment mechanisms demonstrated diverse behaviors with respect to the four analysis criteria. The height-dependant mechanism produced flat, steady state biofilms that lacked sloughing events. Detachment based on substrate limitation produced significant sloughing events. The resulting biofilm structures included distinct, hollow clusters separated by channels. The erosion mechanism produced neither a non-zero steady state nor sloughing events. A mechanism combining all three-detachment mechanisms produced mushroom-like structures. The dynamics of biofilm decay during starvation were distinct for each detachment mechanism. These results show that detachment is a critical determinant of biofilm structure and of the dynamics of biofilm accumulation and loss.


Linking global warming to amphibian declines through its effects on female body condition and survivorship.
There is general consensus that climate change has contributed to the observed decline, and extinction, of many amphibian species throughout the world. However, the mechanisms of its effects remain unclear. A laboratory study in 1980-1981 in which temperate zone amphibians that were prevented from hibernating had decreased growth rates, matured at a smaller size and had increased mortality compared with those that hibernated suggested one possible mechanism. I used data from a field study of common toads (Bufo bufo) in the UK, between 1983 and 2005, to determine whether this also occurs in the field. The results demonstrated two pathways by which global warming may cause amphibian declines. First, there was a clear relationship between a decline in the body condition of female common toads and the occurrence of warmer than average years since 1983. This was paralleled by a decline in their annual survival rates with the relationship between these two declines being highly correlated. Second, there was a significant relationship between the occurrence of mild winters and a reduction in female body size, resulting in fewer eggs being laid annually. Climate warming can, therefore, act on wild temperate zone amphibians by deleteriously affecting their physiology, during and after hibernation, causing increased female mortality rates and decreased fecundity in survivors.


Embers of society: Firelight talk among the Ju/'hoansi Bushmen.
Much attention has been focused on control of fire in human evolution and the impact of cooking on anatomy, social, and residential arrangements. However, little is known about what transpired when firelight extended the day, creating effective time for social activities that did not conflict with productive time for subsistence activities. Comparison of 174 day and nighttime conversations among the Ju/'hoan (!Kung) Bushmen of southern Africa, supplemented by 68 translated texts, suggests that day talk centers on economic matters and gossip to regulate social relations. Night activities steer away from tensions of the day to singing, dancing, religious ceremonies, and enthralling stories, often about known people. Such stories describe the workings of entire institutions in a small-scale society with little formal teaching. Night talk plays an important role in evoking higher orders of theory of mind via the imagination, conveying attributes of people in broad networks (virtual communities), and transmitting the "big picture" of cultural institutions that generate regularity of behavior, cooperation, and trust at the regional level. Findings from the Ju/'hoan are compared with other hunter-gatherer societies and related to the widespread human use of firelight for intimate conversation and our appetite for evening stories. The question is raised as to what happens when economically unproductive firelit time is turned to productive time by artificial lighting.


Association Between Low-Density Lipoprotein Cholesterol (LDL-C) Level and Unfavorable Outcomes in Participants of Ischemic Stroke without Diabetes: A Multi-Center Retrospective Study.
BACKGROUND The impact of low-density lipoprotein cholesterol (LDL-C) levels on outcomes in patients with non-diabetic acute ischemic stroke remains uncertain. The objective of this study was to explore whether LDL-C could refine outcomes after acute ischemic stroke in patients with non-diabetic acute ischemic stroke. MATERIAL AND METHODS A multi-center, retrospective, clinical-based study was conducted within eight hospitals between January 2015 and August 2016. Adjusted odds ratio (aOR) was used for measurement of unfavorable outcome which was evaluated by the modified Rankin Scale (mRS) score at 6 months after acute ischemic stroke, estimated categorically according to multivariate logistic regression. RESULTS A total of 1614 participants with non-diabetic acute ischemic stroke were enrolled, of which 376 patients (23.3%) had unfavorable neurologic outcomes at 6 months. After multivariate analysis comparing 4 LDL-C levels by quartiles (Q), we found that compared to Q1 (LDL-C level ≤2.41 mmol/L), there was a significant association between the frequency of unfavorable outcomes and levels of LDL-C (Q3: 2.95-3.54 mmol/L) for all participants (adjusted odds ratio [aOR]=0.63; 95% CI: 0.44-0.92, P=0.016) and patients with first ever strokes (aOR=0.52; 95% CI: 0.31-0.87, P=0.013). CONCLUSIONS Compared to lower LDL-C levels, non-diabetic patients with LDL-C levels in Q3 (2.95-3.54 mmol/L), were less likely to have unfavorable functional outcomes at 6 months after acute ischemic stroke. Managing HDL-C is one of the most important steps for the recovery of acute ischemic stroke.


Structure and function of the initially transcribing RNA polymerase II-TFIIB complex.
The general transcription factor (TF) IIB is required for RNA polymerase (Pol) II initiation and extends with its B-reader element into the Pol II active centre cleft. Low-resolution structures of the Pol II-TFIIB complex indicated how TFIIB functions in DNA recruitment, but they lacked nucleic acids and half of the B-reader, leaving other TFIIB functions enigmatic. Here we report crystal structures of the Pol II-TFIIB complex from the yeast Saccharomyces cerevisiae at 3.4 Å resolution and of an initially transcribing complex that additionally contains the DNA template and a 6-nucleotide RNA product. The structures reveal the entire B-reader and protein-nucleic acid interactions, and together with functional data lead to a more complete understanding of transcription initiation. TFIIB partially closes the polymerase cleft to position DNA and assist in its opening. The B-reader does not reach the active site but binds the DNA template strand upstream to assist in the recognition of the initiator sequence and in positioning the transcription start site. TFIIB rearranges active-site residues, induces binding of the catalytic metal ion B, and stimulates initial RNA synthesis allosterically. TFIIB then prevents the emerging DNA-RNA hybrid duplex from tilting, which would impair RNA synthesis. When the RNA grows beyond 6 nucleotides, it is separated from DNA and is directed to its exit tunnel by the B-reader loop. Once the RNA grows to 12-13 nucleotides, it clashes with TFIIB, triggering TFIIB displacement and elongation complex formation. Similar mechanisms may underlie all cellular transcription because all eukaryotic and archaeal RNA polymerases use TFIIB-like factors, and the bacterial initiation factor sigma has TFIIB-like topology and contains the loop region 3.2 that resembles the B-reader loop in location, charge and function. TFIIB and its counterparts may thus account for the two fundamental properties that distinguish RNA from DNA polymerases: primer-independent chain initiation and product separation from the template.


Differential expression and cross-talk of peroxisome proliferator-activated receptor γ and retinoid-X receptor α in urothelial carcinomas of the bladder.
the peroxisome proliferator-activated receptor γ (PPARγ), known to play a key role in homeostatic biological pathways, is also implicated in the process of carcinogenesis. Ligands for PPARγ and its heterodimeric partner, retinoid-X receptor (RXR), have exhibited anticancer effects both in vitro and in vivo. Unexpectedly, some studies suggested that PPARγ ligands may stimulate cancer formation. This study aimed to estimate the signaling of PPARγ-RXRα heterodimer in bladder urothelial carcinomas (BUC). we studied PPARγ and RXRα expression in specimens obtained from 97 patients with BUC of various grades and stages using immunohistochemistry. PPARγ expression was significantly downregulated with BUC stage and grade progression, and the dynamics of this phenomenon was significantly influenced by RXRα's level of expression. the positive association of PPARγ expression in BUC with more differentiated, non-invasive tumors is strengthened by the presence of RXRα. This knowledge could probably be of use in the development of new chemotherapeutic agents.


Berardinelli-Seip congenital lipodystrophy 2 regulates adipocyte lipolysis, browning, and energy balance in adult animals.
Mutations in BSCL2/SEIPIN cause Berardinelli-Seip congenital lipodystrophy type 2 (BSCL2), but the mechanisms whereby Bscl2 regulates adipose tissue function are unclear. Here, we generated adipose tissue (mature) Bscl2 knockout (Ad-mKO) mice, in which Bscl2 was specifically ablated in adipocytes of adult animals, to investigate the impact of acquired Bscl2 deletion on adipose tissue function and energy balance. Ad-mKO mice displayed reduced adiposity and were protected against high fat diet-induced obesity, but not insulin resistance or hepatic steatosis. Gene expression profiling and biochemical assays revealed increased lipolysis and fatty acid oxidation in white adipose tissue (WAT) and brown adipose tissue , as well as browning of WAT, owing to induction of cAMP/protein kinase A signaling upon Bscl2 deletion. Interestingly, Bscl2 deletion reduced food intake and downregulated adipose β3-adrenergic receptor (ADRB3) expression. Impaired ADRB3 signaling partially offsets upregulated browning-induced energy expenditure and thermogenesis in Ad-mKO mice housed at ambient temperature. However, this counter-regulatory response was abrogated under thermoneutral conditions, resulting in even greater body mass loss in Ad-mKO mice. These findings suggest that Bscl2 regulates adipocyte lipolysis and β-adrenergic signaling to produce complex effects on adipose tissues and whole-body energy balance.


Favorable Pregnancy Outcome in a Granulomatosis With Polyangiitis Patient With Renal Insufficiency.
To present a case of successful pregnancy outcome in a granulomatosis with polyangiitis (GPA) patient with renal insufficiency. GPA, formerly known as Wegener's granulomatosis, is a rare necrotizing systemic vasculitis, presenting with classical clinical triad of manifestations involving upper and lower airway and glomerulonephritis. An association of Antineutrophil cytoplasmic antibodies with GPA has been established and the antibodies are present in most patients with active disease. Pregnancy with GPA is burdened with the risk of possible maternal and fetal complications, further leading to higher morbidity and mortality rate. Due to sparsity of studies of GPA in pregnancy, management needs to be individualised. Diagnostic workup should include serological markers, radiological and histopathological examination. Cyclophosphamide combined with prednisolone is the standard induction regimen. A 22-year-old woman, multigravida at 35 weeks of gestation was referred to our department owing to 1-year diagnosis of GPA. During active phase, the disease manifested as pneumonia and acute kidney injury and perinuclear anti-neutrophil cytoplasmic antibodies (P-ANCAs) were positive. She received pulse therapy of injection cyclophosphamide and methylprednisolone as induction regimen, followed by tapering doses of oral prednisolone and azathioprine for maintenance therapy. The disease was in remission at the onset of pregnancy but had flare up at 34 - 35 weeks of gestation and she presented with renal dysfunction. Neither the disease nor the treatment adversely affected the pregnancy and she delivered a healthy baby at 37 weeks. The unpredictable disease course and complications at unexpected gestation appears to be a major variable to take into account when assessing the risk of pregnancy with GPA. Early diagnosis, monitoring and timely intervention resulted in favourable pregnancy outcome in our patient.


No efficacy of annual gynaecological screening in BRCA1/2 mutation carriers; an observational follow-up study.
BRCA1/2 mutation carriers are offered gynaecological screening with the intention to reduce mortality by detecting ovarian cancer at an early stage. We examined compliance and efficacy of gynaecological screening in BRCA1/2 mutation carriers. In this multicentre, observational, follow-up study we examined medical record data of a consecutive series of 888 BRCA1/2 mutation carriers who started annual screening with transvaginal ultrasonography and serum CA125 between 1993 and 2005. The women were annually screened for 75% of their total period of follow-up. Compliance decreased with longer follow-up. Five of the 10 incident cancers were interval tumours, diagnosed in women with a normal screening result within 3-10 months before diagnosis. No difference in stage distribution between incident screen-detected and interval tumours was found. Eight of the 10 incident cancers were stage III/IV (80%). Cancers diagnosed in unscreened family members had a similar stage distribution (77% in stage III/IV). The observed number of cases detected during screening was not significantly higher than expected (Standardized Incidence Ratio (SIR): 1.5, 95% confidence interval: 0.7-2.8). For the subgroup that was fully compliant to annual screening, a similar SIR was found (1.6, 95% confidence interval: 0.5-3.6). Despite annual gynaecological screening, a high proportion of ovarian cancers in BRCA1/2 carriers are interval cancers and the large majority of all cancers are diagnosed in advanced stages. Therefore, it is unlikely that annual screening will reduce mortality from ovarian cancer in BRCA1/2 mutation carriers.


In situ STEM study on the morphological evolution of copper-based nanoparticles during high-temperature redox reactions.
Despite the broad relevance of copper nanoparticles in industrial applications, the fundamental understanding of oxidation and reduction of copper at the nanoscale is still a matter of debate and remains within the realm of bulk or thin film-based systems. Moreover, the reported studies on nanoparticles vary widely in terms of experimental parameters and are predominantly carried out using either ex situ observation or environmental transmission electron microscopy in a gaseous atmosphere at low pressure. Hence, dedicated studies in regards to the morphological transformations and structural transitions of copper-based nanoparticles at a wider range of temperatures and under industrially relevant pressure would provide valuable insights to improve the application-specific material design. In this paper, copper nanoparticles are studied using in situ Scanning Transmission Electron Microscopy to discern the transformation of the nanoparticles induced by oxidative and reductive environments at high temperatures. The nanoparticles were subjected to a temperature of 150 °C to 900 °C at 0.5 atm partial pressure of the reactive gas, which resulted in different modes of copper mobility both within the individual nanoparticles and on the surface of the support. Oxidation at an incremental temperature revealed the dependency of the nanoparticles' morphological evolution on their initial size as well as reaction temperature. After the formation of an initial thin layer of oxide, the nanoparticles evolved to form hollow oxide shells. The kinetics of formation of hollow particles were simulated using a reaction-diffusion model to determine the activation energy of diffusion and temperature-dependent diffusion coefficient of copper in copper oxide. Upon further temperature increase, the hollow shell collapsed to form compact and facetted nanoparticles. Reduction of copper oxide was carried out at different temperatures starting from various oxide phase morphologies. A reduction mechanism is proposed based on the dynamic of the reduction-induced fragmentation of the oxide phase. In a broader perspective, this study offers insights into the mobility of the copper phase during its oxidation-reduction process in terms of microstructural evolution as a function of nanoparticle size, reaction gas, and temperature.


Short-term resistance training with blood flow restriction enhances microvascular filtration capacity of human calf muscles.
Resistance training increases muscle strength and endurance but may require high intensity and long duration to enhance capillarity. Vascular occlusion during low-load resistance training augments the strength and endurance gains compared with low-load resistance training alone, but in this study we investigated whether it also promotes microvascular filtration capacity, an index of capillarity. Nine healthy males performed short-term low-intensity resistance training of the calf muscles (four sets of 50 heel raises, three times a week for 4 weeks) under restricted (thigh cuff inflated to 150 mmHg on the non-dominant leg) or unrestricted (dominant leg without thigh cuff) blood flow conditions. Before and after resistance training, calf filtration capacity and resting blood flow were assessed by strain gauge plethysmography, and calf muscle strength and fatigue were assessed respectively by maximal voluntary contraction and force decline during electrically evoked ischaemic contractions in both legs. Calf filtration capacity increased by 26% in the restricted leg but did not increase significantly in the unrestricted leg. Calf muscle strength was 18% greater in the restricted leg but unchanged in the unrestricted leg. Calf muscle fatigue and resting blood flow did not change in either leg. Resistance training promoted microvascular filtration capacity, an effect that was somewhat enhanced by blood flow restriction, and could be due to increased capillarization.


Age-related somatic structural changes in the nuclear genome of human blood cells.
Structural variations are among the most frequent interindividual genetic differences in the human genome. The frequency and distribution of de novo somatic structural variants in normal cells is, however, poorly explored. Using age-stratified cohorts of 318 monozygotic (MZ) twins and 296 single-born subjects, we describe age-related accumulation of copy-number variation in the nuclear genomes in vivo and frequency changes for both megabase- and kilobase-range variants. Megabase-range aberrations were found in 3.4% (9 of 264) of subjects ≥60 years old; these subjects included 78 MZ twin pairs and 108 single-born individuals. No such findings were observed in 81 MZ pairs or 180 single-born subjects who were ≤55 years old. Recurrent region- and gene-specific mutations, mostly deletions, were observed. Longitudinal analyses of 43 subjects whose data were collected 7-19 years apart suggest considerable variation in the rate of accumulation of clones carrying structural changes. Furthermore, the longitudinal analysis of individuals with structural aberrations suggests that there is a natural self-removal of aberrant cell clones from peripheral blood. In three healthy subjects, we detected somatic aberrations characteristic of patients with myelodysplastic syndrome. The recurrent rearrangements uncovered here are candidates for common age-related defects in human blood cells. We anticipate that extension of these results will allow determination of the genetic age of different somatic-cell lineages and estimation of possible individual differences between genetic and chronological age. Our work might also help to explain the cause of an age-related reduction in the number of cell clones in the blood; such a reduction is one of the hallmarks of immunosenescence.


Cerebral blood flow in chronic symptomatic mild traumatic brain injury.
Patients with mild traumatic brain injury (MTBI) challenge physicians' skills and test their patience. Their manifold symptomatology is often not supported by objective neurological findings. We sought to compare regional cerebral blood flow (rCBF) between symptomatic subjects with longstanding MTBI and healthy controls, and to examine the correspondence between neuropsychological deficit and rCBF compromise. Twenty-eight clinically symptomatic male subjects with MTBI and twenty matched controls underwent neuropsychological testing and Tc-99m-HMPAO brain SPECT imaging. Neuropsychological test data were used to categorize subjects into sub-groups according to the presumed location of lesions based on their neurobehavioral deficits. Image subtraction comparisons were made between controls, all MTBI subjects and sub-groups. MTBI patients demonstrated regions of hypoperfusion in frontal, pre-frontal and temporal cortices, and sub-cortical structures. Hypoperfusion in 'frontal', 'left posterior' and to a lesser extent 'sub-cortical' sub-groups was concordant with neuropsychological localization. This was not the case for the 'right posterior' group, where no concordance was found. The rCBF is reduced in symptomatic patients with longstanding MTBI and unremarkable structural brain imaging. Although group analysis is appropriate for the generation of statistically significant differences, the clinical application of brain SPECT imaging in MTBI calls for a capability to associate clinical examination, neuropsychological assessment and cerebral perfusion at the individual subject level. Such competence is still to be attained.


Social desirability effects in driver behavior inventories.
The use of lie scales to control for common method variance in driver behavior inventories has been very limited. Given that such questionnaires often use self-reported safety variables as criteria, and have social implications, the risk of artefactual associations is high. A questionnaire containing scales from several well known driver inventories that have been claimed to predict traffic accident involvement was distributed three times to a group of young drivers in a driver education program, as well as a random group twice. The Driver Impression Management scale (DIM) was used to control for socially desirable responding. For all behavior scales, the correlation with the DIM scale was substantial. If a scale correlated with self-reported crashes, the amount of predictive power was more than halved when social desirability was controlled for. Results were similar for both samples and all waves. The predictive power of the behavior scales was not increased when values were averaged over questionnaire waves, as should have been the case if the measurement and predictive power were valid. Results were similar for self-reported penalty points. The present results indicate that even the most well-known and accepted psychometric scales used in driver research are susceptible to social desirability bias. As social desirability is only one of a number of common method variance mechanisms that can create artefactual associations, and the great popularity of the self-report methodology, the problem for traffic research is grave. Organizations that fund traffic safety research need to re-evaluate their policies regarding what methods are acceptable. The use of self-reported independent and dependent variables can lead to directly misleading results, with negative effects on traffic safety.


Associations of occupational standing with musculoskeletal symptoms: a systematic review with meta-analysis.
Given the high exposure to occupational standing in specific occupations, and recent initiatives to encourage intermittent standing among white-collar workers, a better understanding of the potential health consequences of occupational standing is required. We aimed to review and quantify the epidemiological evidence on associations of occupational standing with musculoskeletal symptoms. A systematic review was performed. Data from included articles were extracted and described, and meta-analyses conducted when data were sufficiently homogeneous. Electronic databases were systematically searched. Peer-reviewed articles on occupational standing and musculoskeletal symptoms from epidemiological studies were identified. Of the 11 750 articles screened, 50 articles reporting 49 studies were included (45 cross-sectional and 5 longitudinal; n=88 158 participants) describing the associations of occupational standing with musculoskeletal symptoms, including low-back (39 articles), lower extremity (14 articles) and upper extremity (18 articles) symptoms. In the meta-analysis, 'substantial' (>4 hours/workday) occupational standing was associated with the occurrence of low-back symptoms (pooled OR (95% CI) 1.31 (1.10 to 1.56)). Evidence on lower and upper extremity symptoms was too heterogeneous for meta-analyses. The majority of included studies reported statistically significant detrimental associations of occupational standing with lower extremity, but not with upper extremity symptoms. The evidence suggests that substantial occupational standing is associated with the occurrence of low-back and (inconclusively) lower extremity symptoms, but there may not be such an association with upper extremity symptoms. However, these conclusions are tentative as only limited evidence was found from high-quality, longitudinal studies with fully adjusted models using objective measures of standing.


Femoral medullary infarction secondary to canine total hip arthroplasty.
To evaluate the prevalence of femoral intramedullary infarction after total hip arthroplasty (THA) and to determine whether any specific femoral morphology predisposes to bone infarction. Retrospective clinical study. All dogs from our hospital population undergoing THA between 1984 and 1997 with radiographic follow-up available at 1 year or more postoperatively. A case control study was conducted within the THA group to determine risk factors predisposing to femoral infarction after THA. Medical records and radiographs were reviewed. Data were collected on clinical parameters, femoral morphology, prosthesis, and bone changes. Radiographic diagnosis was confirmed using histopathology in 11 femora. Radiographs of 50 age-matched control dogs weighing more than 20 kg with coxofemoral degenerative joint disease were randomly chosen to determine the prevalence of bone infarction in nonoperated dogs. Ninety-one dogs with 110 THA were included in the study. Fifteen of the 110 femora with THA had radiographic evidence of infarction (14%). Infarction was not present in any femora in the control group. There was no significant difference in the prevalence of infarction between dogs that received cemented or uncemented prostheses. Clinical signs were not reported in any patient that developed femoral infarction. Young age (P = .03) and a distance between the greater trochanter and nutrient foramen greater than 79 mm (P = .008) predisposed dogs to femoral infarction. Over time, three infarcts decreased in size radiographically, five remained unchanged, and three expanded. An osteosarcoma developed at the site of a bone infarct in one dog. Femoral intramedullary infarction occurred in 15 of 110 THA. Young age at the time of THA and a greater distance between the greater trochanter and the nutrient foramen predisposed to infarction. Intramedullary infarction occurs after canine THA. These bone infarcts do not appear to cause clinical signs; however, they may present a diagnostic challenge. Malignant transformation could potentially result from medullary infarction.


Learning from number board games: you learn what you encode.
We tested the hypothesis that encoding the numerical-spatial relations in a number board game is a key process in promoting learning from playing such games. Experiment 1 used a microgenetic design to examine the effects on learning of the type of counting procedure that children use. As predicted, having kindergartners count-on from their current number on the board while playing a 0-100 number board game facilitated their encoding of the numerical-spatial relations on the game board and improved their number line estimates, numeral identification, and count-on skill. Playing the same game using the standard count-from-1 procedure led to considerably less learning. Experiment 2 demonstrated that comparable improvement in number line estimation does not occur with practice encoding the numerals 1-100 outside of the context of a number board game. The general importance of aligning learning activities and physical materials with desired mental representations is discussed.


Sphingosine kinase inhibitors and cancer: seeking the golden sword of Hercules.
There is considerable evidence that sphingosine kinases play a key role in cancer progression, which might involve positive selection of cancer cells that have been provided with a survival and growth advantage as a consequence of overexpression of the enzyme. Therefore, inhibitors of sphingosine kinase represent a novel class of compounds that have potential as anticancer agents. Poor inhibitor potency is a major issue that has precluded successful translation of these compounds into the clinic. However, recent discoveries have shown that sphingosine kinase 1 is an allosteric enzyme and that some inhibitors offer improved effectiveness by inducing proteasomal degradation of the enzyme or having nanomolar potency. Herein, we provide a perspective about these recent developments and highlight the importance of translating basic pharmacologic and biochemical findings on sphingosine kinase into new drug discovery programs for treatment of cancer.


Procalcitonin as a marker of serious bacterial infections in febrile children younger than 3 years old.
There is no perfectly sensitive or specific test for identifying young, febrile infants and children with occult serious bacterial infections (SBIs). Studies of procalcitonin (PCT), a 116-amino-acid precursor of the hormone calcitonin, have demonstrated its potential as an acute-phase biomarker for SBI. The objective of this study was to compare performance of serum PCT with traditional screening tests for detecting SBIs in young febrile infants and children. This was a prospective, multicenter study on a convenience sample from May 2004 to December 2005. The study was conducted in four emergency departments (EDs): one pediatric ED and three EDs with pediatric units, all with academic faculty on staff. A total of 226 febrile children 36 months old or younger who presented to the four participating EDs and were evaluated for SBI by blood, urine, and/or cerebral spinal fluid (CSF) cultures were included. The test characteristics (with 95% confidence intervals [CIs]) of the white blood cell (WBC) counts including neutrophil and band counts were compared with PCT for identifying SBI. Thirty children had SBIs (13.3%, 95% CI = 8.85 to 17.70). Four (13.3%) had bacteremia (including one with meningitis), 18 (60.0%) had urinary tract infections (UTIs), and eight (26.6%) had pneumonia. Children with SBIs had higher WBC counts (18.6 × 10(9) ± 8.6 × 10(9) cells/L vs. 11.5 × 10(9) ± 5.3 × 10(9) cells/L, p < 0.001), higher absolute neutrophil counts (ANCs; 10.6 × 10(9) ± 6.7 × 10(9) cells/L vs. 5.6 × 10(9) ± 3.8 × 10(9) cells/L, p = 0.009), higher absolute band counts (0.90 × 10(9) ± 1.1 × 10(9) cells/L vs. 0.35 × 10(9) ± 0.6 × 10(9) cells/L, p = 0.009), and higher PCT levels (2.9 ± 5.6 ng/mL vs. 0.4 ± 0.8 ng/mL, p = 0.021) than those without SBIs. In a multivariable logistic regression analysis, the absolute band count and PCT were the two screening tests independently associated with SBI, although the area under the receiver operating characteristic (ROC) curve for PCT was the largest (0.80, 95% CI = 0.71 to 0.89). Procalcitonin is a more accurate biomarker than traditional screening tests for identifying young febrile infants and children with serious SBIs. Further study on a larger cohort of young febrile children is required to definitively determine the benefit of PCT over traditional laboratory screening tests for SBIs.


Typhlitis; is it just in immunocompromised patients?
Typhlitis is necrotizing inflammation of the cecum manifested by febrile right lower quadrant abdominal pain. All cases in literature have been described in immunocompromised such patients with hematological or solid malignancies, patients with neutropenia, patients with AIDS, and patients following immunosuppressive therapy for transplants. We report a case of a 72 year old non-immunocompromised patient who developed fever, nausea, vomiting and severe right lower quadrant pain. Computed Tomography (CT) of the abdomen and pelvis showed circumferential thickening of the cecum as well as inflammatory stranding of the adjacent mesenteric fat. The patient was treated with fluids and antibiotics. The patient's symptoms resolved over a period of few days. We believe this case represents an example of typhlitis in non-immunocompromised patient, the first such case reported. Therefore, typhlitis may present in elderly patients even in the absence of neutropenia or immunosuppression.


A Bayesian method to estimate the depth and the range of phonating sperm whales using a single hydrophone.
Some bioacousticians have used a single hydrophone to calculate the depth/range of phonating diving animals. The standard one-hydrophone localization method uses multipath transmissions (direct path, sea surface, and seafloor reflections) of the animal phonations as a substitute for a vertical hydrophone array. The standard method requires three multipath transmissions per phonation. Bioacousticians who study foraging sperm whales usually do not have the required amount of multipath transmissions. However, they usually detect accurately (using shallow hydrophones towed by research vessels) direct path transmissions and sea surface reflections of sperm whale phonations (clicks). Sperm whales emit a few thousand clicks per foraging dive, therefore researchers have this number of direct path transmissions and this number of sea surface reflections per dive. The author describes a Bayesian method to combine the information contained in those acoustic data plus visual observations. The author's tests using synthetic data show that the accurate estimation of the depth/range of sperm whales is possible using a single hydrophone and without using any seafloor reflections. This method could be used to study the behavior of sperm whales using a single hydrophone in any location no matter what the depth, the relief, or the constitution of the seafloor might be.


The Quest for System-Theoretical Medicine in the COVID-19 Era.
Precision medicine and molecular systems medicine (MSM) are highly utilized and successful approaches to improve understanding, diagnosis, and treatment of many diseases from bench-to-bedside. Especially in the COVID-19 pandemic, molecular techniques and biotechnological innovation have proven to be of utmost importance for rapid developments in disease diagnostics and treatment, including DNA and RNA sequencing technology, treatment with drugs and natural products and vaccine development. The COVID-19 crisis, however, has also demonstrated the need for systemic thinking and transdisciplinarity and the limits of MSM: the neglect of the bio-psycho-social systemic nature of humans and their context as the object of individual therapeutic and population-oriented interventions. COVID-19 illustrates how a medical problem requires a transdisciplinary approach in epidemiology, pathology, internal medicine, public health, environmental medicine, and socio-economic modeling. Regarding the need for conceptual integration of these different kinds of knowledge we suggest the application of general system theory (GST). This approach endorses an organism-centered view on health and disease, which according to Ludwig von Bertalanffy who was the founder of GST, we call Organismal Systems Medicine (OSM). We argue that systems science offers wider applications in the field of pathology and can contribute to an integrative systems medicine by (i) integration of evidence across functional and structural differentially scaled subsystems, (ii) conceptualization of complex multilevel systems, and (iii) suggesting mechanisms and non-linear relationships underlying the observed phenomena. We underline these points with a proposal on multi-level systems pathology including neurophysiology, endocrinology, immune system, genetics, and general metabolism. An integration of these areas is necessary to understand excess mortality rates and polypharmacological treatments. In the pandemic era this multi-level systems pathology is most important to assess potential vaccines, their effectiveness, short-, and long-time adverse effects. We further argue that these conceptual frameworks are not only valid in the COVID-19 era but also important to be integrated in a medicinal curriculum.


Rapid identification of metallo- and serine beta-lactamases.
Simple methods to detect, identify, and differentiate metallo- and serine beta-lactamases were developed and used to differentiate enzymes produced by 17 clinical isolates of Xanthomonas maltophilia. All isolates exhibited beta-lactamase activity, and in 16 strains this was induced by imipenem. All but one isolate hydrolyzed imipenem (and meropenem), and in all cases this activity was inhibited by 1 mM EDTA. The metallo- and serine beta-lactamases in the cell extracts were distinguished on isoelectric focusing (IEF) gels by using the following procedures. (i) Cell lysates were preincubated with 83 mM EDTA prior to IEF and subsequent visualization with nitrocefin, and (ii) after IEF, the gels were overlaid with either 1 mM zinc sulfate or 100 microM BRL 42715 before staining with nitrocefin. Bands of beta-lactamase activity which were removed by BRL 42715 but unaffected by EDTA or zinc sulfate were categorized as serine beta-lactamases. Bands which were unaffected by BRL 42715 but inhibited by EDTA or enhanced by zinc sulfate were classified as metallo-beta-lactamases. By using this approach, seven metallo-beta-lactamases were differentiated with pI values of 4.8 (two strains), 5.5 (four strains), 5.7 (one strain), 6.0 (one strain), 6.4 (four strains), 6.6 (one strain), and 6.8 (three strains). The metallo-beta-lactamase band with a pI of 6.4 aligned with the recently characterized metallo-beta-lactamase from X. maltophilia 511. Heterogeneity was also observed for the serine beta-lactamases: 14 isolates elaborated serine beta-lactamase activity which focused with major bands with at least eight different pIs. The remaining three strains produced serine beta-lactamases which focused with five distinct bands with pIs of 6.4, 6.2, 5.7, 5.5, and 5.2. We conclude that X. maltophilia produces many types of metallo- and serine beta-lactamases distinguishable by these new methods and that the previously reported L-1 and L-2 enzymes are not solely representative of the beta-lactamases produced by this species.