Site-Specific Turn-On Fluorescent Labeling of DNA-Interacting Protein Using Oligodeoxynucleotides That Modify Lysines To Produce 5,6-Dimethoxy 3-Methyleneisoindolin-1-one.
We have developed oligodeoxynucleotides (ODNs) that modify primary amines to produce 5,6-dimethoxy 3-methyleneisoindolin-1-one. Compared to the oxygen isosteric fluorophore, 4,5-dimethoxyphthalimide, this methyleneisoindolinone was more stable and exhibited an 85 nm blue-shifted fluorescent emission (λmax at 425 nm) with an intensity comparable to that of the phthalimide. Reaction of the DNA-binding domain of Escherichia coli DnaA protein with an ODN containing its binding sequence efficiently afforded a modified fluorescent protein at a specific lysine residue in the proximity of the ODN. A full-length DnaA protein was also successfully fluorescently labeled. These results demonstrate the potential utility of the ODNs developed in this study for the fluorescent labeling of DNA-interacting protein at the lysine residue of interest.


Epigenetic drugs induce the potency of classic chemotherapy, suppress post-treatment re-growth of breast cancer, but preserve the wound healing ability of stem cells.
Epigenetic therapy augments neoadjuvant chemotherapy (NACT) in breast cancer and may aid post-surgical wound healing affected by NACT. Our study investigates: (1) The cytotoxicity of classic paclitaxel chemotherapy on triple negative breast cancer (TNBC) independently and in combination with epigenetic drugs. (2) The sustainable inhibition of breast cancer regrowth following paclitaxel and epigenetic therapies. (3) The effects of paclitaxel with and without epigenetic therapy on the post-treatment viability and wound healing potential of adipose stem cells (ASCs). Cytotoxicity assays were performed on TNBC and ASCs. Cells were treated and recovered in drug-free medium. Cell viability was measured via cell counts and MTT assays. W -ound healing was tested with scratch assays. The combination of epigenetic drugs shows increased toxicity against TNBC cells compared to standard chemotherapy alone. Moreover, the combination of paclitaxel with epigenetic treatments causes cancer toxicity that is sustainable to TNBC cells after the drugs' removal with minimal effect on ASCs wound healing ability. The use of epigenetic drugs in addition to standard chemotherapy is cytotoxic to TNBC cells and prevents post-treatment recovery of TNBC while maintaining ASC wound healing ability. This strategy may be useful in maximizing post-surgical wound healing following NACT in TNBC.


Increased circulating endothelial cells in acute heart failure: comparison with von Willebrand factor and soluble E-selectin.
Circulating endothelial cells (CECs) in the peripheral blood, probably representing the most direct evidence of endothelial cell damage, are increased in myocardial infarction, unstable angina and critical limb ischaemia. As chronic heart failure is also associated with endothelial abnormalities, we hypothesised that CECs are raised in acute heart failure and that they would correlate with plasma indices of endothelial perturbation, that is, von Willebrand factor (vWf) and soluble E-selectin. We studied 30 patients with acute heart failure (venesected within 24 h of emergency hospital admission), 30 patients with chronic stable heart failure (venesected as out-patients, all patients in sinus rhythm with ejection fraction < or = 40%) and 20 healthy controls. CECs were quantified using epifluorescence microscopy after CD146-immunomagnetic separation and phenotyped by streptavidin/biotin immunocytochemistry. Citrated plasma was analysed for soluble E-selectin and vWf by ELISA. Levels of CECs, vWf and soluble E-selectin were significantly higher (all p<0.01) in patients with heart failure compared to controls, with no significant differences between acute and chronic heart failure. CECs correlated with plasma vWf (p<0.0001) and soluble E-selectin (p = 0.022) but not ejection fraction or NYHA class. In multiple regression analysis, heart failure was the only independent predictor of raised CECs (p<0.0001). Immunoperoxidase-defined surface expression of CD34, CD45 and CD36 by CECs was <2%, 0% and 8%, respectively. CECs, a possibly heterologous population, may be used as a novel measure of endothelial damage in acute heart failure and may have implications for the thrombotic risk associated with acute and chronic heart failure and prognosis in this condition.


Factors influencing patients' receptiveness to evidence-based recommendations during the clinical encounter.
To identify the factors that promote or interfere with evidence-based clinical decisions from the patient perspective. We developed four hypothetical scenarios with clinical decisions, such as whether to pursue testing for a chronic condition. We conducted eight focus groups to better understand the influences on individuals' decisions in the context of the scenarios. A patient's relationship with a physician emerged as the strongest influence on whether participants would accept or reject a physician's recommendation. Various patient characteristics and a patient's financial capacity were also important influences. Our findings point to the potential for interventions that improve communications and relationships between physicians and patients to promote evidence-based care.


Recombinant Mycobacterium smegmatis expressing Hsp65-hIL-2 fusion protein and its influence on lymphocyte function in mice.
To construct a strain of recombinant Mycobacterium smegmatis expressing the heat shock protein 65 (Hsp65) and human interleukin 2 (IL-2) fusion protein (rMS-Hsp65/IL-2) and to explore the effect of this construct on lymphocyte function in mice. The fusion gene encoding Hsp65-hIL-2 was cloned into shuttle vector pSMT3. The recombinant plasmid pSMT3-Hsp65-hIL-2 was transferred to Mycobacterium smegmatis by electroporation. Positive clones were selected by hygromycin and identified by PCR. The expression of fusion protein Hsp65-hIL-2 was verified using indirect immunofluorescence staining. Mice were immunized for two times by subcutaneously injection with 1×10(6) CFU rMS-Hsp65/IL-2 at a three-week interval. Two weeks after the second immunization, mice were sacrificed and the serum samples were collected for determination of anti-Hsp65 specific IgG. Splenic lymphocytes were isolated and treated with the rMS-Hsp65/IL-2 to determine lymphocytic proliferation activity by MTT assay. IFN-γ- and IL-2 in the medium of the treated cells were also determined by ELISA. Successful construction of rMS-Hsp65/IL-2 was verified by PCR and immunofluorescence staining. Compared to the splenic lymphocytes isolated from mice immunized with Bacille Calmette-Guerin or mice immunized with Mycobacterium smegmatis alone, the splenic lymphocytes isolated from mice immunized with rMS-Hsp65/IL-2 showed a marked increase in the proliferation of lymphocytes, together with an increased production of important cytokines such as IFN-γ-and IL-2. rMS-Hsp65/IL-2 markedly enhances lymphocyte function. Therefore, the fusion protein generated by rMS-Hsp65/IL-2 may be of potential value in generating an effective vaccine against tuberculosis.


Measurement of depression in patients with chronic obstructive pulmonary disease (COPD).
To estimate the validity of the Hamilton Depression Scale (HDS) in a population of patients with chronic obstructive pulmonary disease (COPD). Forty-nine patients with moderate to severe COPD were examined using the ICD-10 criteria for depression. The mean age of the patients was 71 years and 33 (64%) were women. Forty-six (94%) of the patients were also evaluated using the 17-item HDS including the six-item Hamilton Depression subscale (HDSS). Internal and external validity were measured using factor analysis, Cronbach Coefficient alpha, Loevinger coefficient of homogeneity, correlation analysis and ROC-curves. Twenty-three (47%) of the patients were depressed according to the ICD-10 criteria for depression. The HDSS but not the HDS showed a good internal validity. An acceptable external validity was furthermore shown for the HDSS. The HDSS can be recommended as a suitable depression rating scale for COPD patients.


Increased prevalence of subclinical atherosclerosis in patients with hidradenitis suppurativa (HS).
Chronic inflammatory diseases have been associated with increased prevalence of subclinical atherosclerosis. Hidradenitis suppurativa (HS) is a chronic inflammatory disease involving intertriginous skin. We sought to investigate the potential association between HS and subclinical atherosclerosis. This study included 68 patients with HS and 136 age- and sex-matched healthy control subjects. Patients with history of cardiovascular events, diabetes mellitus, chronic kidney disease, or another concomitant inflammatory condition were excluded. Carotid intima-media thickness and carotid plaques were measured by carotid ultrasonography. Adjustments were made for age, sex, and traditional cardiovascular risk factors. Patients had greater carotid intima-media thickness values than control subjects (0.615 ± 0.097 vs 0.578 ± 0.098 mm; P = .012). Carotid plaques were also more frequent in patients than in control subjects (30.9% vs 22.1%). In the multivariable regression model adjusted for age, sex, and traditional cardiovascular risk factors, HS was significantly related to the presence of carotid plaques (odds ratio 2.99, 95% confidence interval 1.26-7.13; P = .013). Causality could not be assessed. These results indicate an increased frequency of subclinical atherosclerosis in patients with HS. Accordingly, HS should be considered a disease associated with potentially increased cardiovascular risk.


Multiparametric imaging-based differentiation of lymphoma and glioblastoma: using T1-perfusion, diffusion, and susceptibility-weighted MRI.
To compare the diagnostic performance of T1 perfusion magnetic resonance imaging (MRI), diffusion-weighted imaging (DWI), and susceptibility-weighted imaging (SWI) for differentiating primary central nervous system lymphoma (PCNSL) and glioblastoma (GBM). This retrospective study comprised a cohort of 70 patients with glioblastoma and 30 patients with PCNSL. T1 perfusion MRI-derived rCBV_corr (leakage corrected relative cerebral blood volume), apparent diffusion coefficient (ADC) derived from DWI, and intratumoural susceptibility signals intensity (ITSS) measured on SWI were evaluated in these 100 patients. The Mann-Whitney U-test was used for pairwise comparison between groups. The diagnostic performance for differentiating PCNSL from glioblastoma was evaluated by using univariate and multivariable logistic regression analyses and receiver operating characteristic (ROC) analysis. Minimum ADC, maximum rCBVs_corr, kep (back flux exchange rate), and ITSS scores were significantly lower in patients with PCNSL than in those with glioblastoma (p<0.05). On ROC analysis, ITSS showed the best discrimination ability for differentiation of GBM and PCNSL with an area under the ROC curve (AUC) of 0.80. rCBV_corr and ADC showed AUCs of 0.68 and 0.63, respectively. Multiparametric assessment using ADC, rCBV_corr, kep, and ITSS scores significantly increased the diagnostic ability for differentiating PCNSL from GBM as compared to mean ADC, mean rCBV_corr, and ITSS alone or a combination of these parameters. The multiparametric model could correctly discriminate 84% of tumours with a sensitivity and specificity of 90% and 70% with an AUC of 0.92. Multiparametric MRI evaluation using DWI, T1 perfusion MRI, and SWI enabled reliable differentiation of PCNSL and GBM in the majority patients, and these results support an integration of advanced MRI techniques for the diagnostic work-up of patients with these tumours.


[Student program, congress grants and their potential influence on surgical career selection].
Due to a decreased interest in surgical training the German Society of Surgery (GSS) has provided a specially sponsored student program at its annual clinical congress for many years in order to increase the number of applicants for a career in surgery. It remains unclear if this goal is reached because an evaluation has not yet been performed. At the clinical congress in 2014, 200 medical students participated in a special student program with the aid of a congress grant totaling 100,000 €. By means of a questionnaire it was asked how many of the participants in this special program have started a residency in surgery. Almost 17% of the participants of the student program answered and 13 students (6.5% of all participants) stated that they had started a surgical residency. All of these students claimed an interest in surgery even before participation in the congress. Those students who had not yet decided what kind of residency they should choose did not vote for surgery despite their visit to the congress. The student program and the congress grant at the annual clinical congress of the GSS do not result in an increased number of applicants for a residency in surgery. Both incentives might be not more than a strategic marketing instrument. An increased number of medical students applying for a residency in surgery is only to be expected if social developments (e. g. attention to a work-life balance) and their concomitant challenges are reflected in surgical day to day work.


Assessment in basic science instruction: directions for practice and research.
In this essay, we discuss assessment of students' understanding of the basic biomedical sciences during the basic science component of medical education. Because we strongly believe that assessment should follow from and be congruent with course and curricular goals, the first section discusses recent trends in basic science education: where it's been and where it appears to be headed. The second section discusses considerations in the design of an assessment system that reflects curricular goals and encourages students to achieve them; it also identifies some research issues that we think merit attention. In the last section, we outline some of the special assessment issues and problems that arise in the context of problem-based learning (PBL) and discuss the merits of some of these novel assessment approaches, as well as the use of more traditional methods in PBL curricula.


Modification of endoplasmic reticulum Ca2+ stores by select oxidants produces changes reminiscent of those in cells from patients with Alzheimer disease.
Abnormalities in calcium homeostasis and oxidative processes occur in fibroblasts from patients with Alzheimer disease (AD) and in fibroblasts and neurons from transgenic mice bearing a presenilin-1 (PS-1) mutation. Bombesin-releasable endoplasmic reticulum Ca2+ stores (BRCS) are exaggerated in all of these cells. Our previous studies show that H2O2 exaggerates BRCS. The goal of the present study was to determine whether select reactive species exaggerate BRCS in cultured human fibroblasts and to determine if the ability of fibroblasts to handle these specific oxidant species is altered in cells from AD patients. Two fluorescent indicators were used to distinguish different reactive oxygen species (ROS): 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester) (c-DCF) and 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM). ROS were produced by a variety of oxidants, including tert-butyl-hydroxyperoxide (t-BHP), hypoxanthine/xanthine oxidase, S-nitroso-N-acetylpenicillamine, 3-morpholinosydnonimine, and sodium nitroprusside. Different oxidants selectively induced various ROS in distinct patterns. These oxidants also induced selective modification in [Ca2+]i and/or BRCS. Of the several oxidants tested, t-BHP was most specific for exaggerating BRCS without affecting basal [Ca2+]i and inducing only c-DCF-detectable ROS. On the other hand, the results show that NO that reacted with DAF-FM was not responsible for alterations in BRCS. Furthermore, the c-DCF-detectable ROS production induced by t-BHP was higher in fibroblasts from AD patients bearing a PS-1 mutation (n = 7) than in those from aged controls (n = 8). The higher production of c-DCF-detectable ROS may underlie the exaggeration of BRCS in fibroblasts from AD patients. Thus, these results are consistent with the hypothesis that abnormalities in selective cellular ROS cause AD-related changes in intracellular calcium regulation.


[Out-of-hospital ventilation after prolonged weaning].
An increasing number of patients require prolonged weaning from mechanical ventilation as a result of advanced age, patient comorbidities, technical progress in surgery and intensive care medicine. The data of the WeanNet register show that more than half (64%) of patients transferred from the intensive care unit (ICU) to a specialized weaning center could definitely be weaned from the respirator. Weaning failure was associated with prolonged ventilation prior to transfer to a weaning center, low body mass index, pre-existing neuromuscular diseases and advanced age. The number of patients with out of hospital ventilation who had to be re-hospitalized because of ventilation control or as part of emergency management quadrupled in Germany between 2006 and 2016. Invasive out-of-hospital ventilation and long-term noninvasive ventilation are associated with a significant loss of autonomy and with low quality of life. Therefore, the initiation must be carefully reviewed and regularly re-evaluated in the context of patient comorbidities and, if necessary, decisions should be made with respect to changing treatment targets. Specialized weaning centers have been established for patients in whom weaning on the ICU was unsuccessful. In cases of persisting weaning failure the adequate transition to out-of-hospital ventilation should be managed by a weaning unit. Weaning centers are responsible for outpatient invasive or noninvasive ventilation strategies and control of treatment quality. Depending on the infrastructure and networking of the respective weaning center, it is basically also possible to provide outpatient care for clinically stable patients in a cooperation model together with pulmonologists in private practice experienced in respiratory medicine.


Chemical incident planning: a review of the literature.
Chemical incidents cause problems for Accident and Emergency (A & E) departments which are different from those recognized in other major accidents. As well as possible trauma there is the added problem of contamination. A & E departments must be prepared for chemical disasters with a chemical incident plan, decontamination facilities and protective clothing for all staff involved. The plan should include how to protect the hospital from contamination and how to prevent its personnel from becoming secondary casualties. The results of a survey into the preparedness of inner London A & E departments were published in issue 6.2 of this journal, April 1998. On 20 March, 1995 a religious cult released a nerve gas (sarin) into the Tokyo subway system. More than 5500 people needed hospital treatment and 11 people died (Reuter News Service, 22 March, 1995). The hospitals were overwhelmed with casualties. Once chemical exposure of victims was suspected, clothing was removed and patients were showered. Stretcher patients were decontaminated by means of bed bathing and a change of bedclothes (Okumura et al 1996). Hospital contamination was a problem during this incident due to the delay in recognising chemical exposure. This resulted in staff contamination. Many lessons were learned from this disaster, including the need to be prepared for all eventualities. If a similar incident occurred in the London Underground system, it would be catastrophic. Underground staff, police, fire fighters, ambulance staff and the general public could all be contaminated before the chemical was identified. There could be mass casualties, including personnel in the emergency services. How would London A & E departments cope, faced with a chemical disaster such as this?


Comparison of digital and conventional assessment methods for a single tooth preparation and educational satisfaction.
Digital feedback for tooth preparation can provide dental practitioners with more objective and accurate evaluations compared to conventional methods. This study aimed to evaluate the educational effect and satisfaction of digital feedback compared with those of the conventional putty index method for tooth preparation. Forty-eight third-grade dental students were selected. All students performed a full-coverage preparation on a right mandibular first molar resin tooth. They were randomly divided into four groups (n = 12)-no guide (control), putty index, digital feedback, and digital and putty index assessment. Three-dimensional analysis was performed using an analysis software (GomInspect 2018, Gom) to evaluate the amount of tooth structure removed. At the end of the practice, the students completed a questionnaire to evaluate the educational satisfaction of the respective methods. There was no statistical significance of the amount of preparation amongst groups in most of the measured areas only except for several specific points. Overall occlusal surface showed 0.99 ± 0.27 mm in the N group (no guide) and 1.15 ± 0.31 mm in the D group (digital feedback), and overall axial surface showed no statistical differences (p > .05). The groups that used digital assessment showed a high level of satisfaction compared with conventional assessment. It was difficult to confirm that the digital-based feedback promotes accurate tooth preparation compared to conventional feedback within the limitation of this study. However, it improved educational satisfaction and permitted objective evaluation.


Prediction of postural risk of fall initiation based on a two-variable description of body dynamics: position and velocity of center of mass.
This research addresses the question: what is the risk of fall initiation at a certain human posture? There are postures from which no one is able to keep their balance and a fall will surely initiate (risk=1), and others from which everyone may regain their stability (risk=0). In other postures, only a portion of people can control their stability. One may interpret risk to chance of a fall to be initiated, and based on the portion of fallers assign a risk value to a given human posture (postural risk). Human posture can be mapped to a point in a 2-dimensional space: the x-v plane, the axes of which are horizontal components of the position and velocity of the center of mass of the body. For every pair of (x, v), the outcome of the balance recovery problem defines whether a person with a given strength level is able to regain their stability when released from a posture corresponding to that point. Using strength distribution data, we estimated the portion of the population who will initiate a fall if starting at a certain posture. A fast calculation approach is also introduced to replace the time-consuming method of solving the recovery problem many times. Postural risk of fall initiation for situations expressed by (x, v) pairs for the entire x-v plane is calculated and shown in a color-map.


Bacterial response to formaldehyde in an MFC toxicity sensor.
Microbial fuel cells (MFCs) have been extensively studied as toxicity sensors. MFCs have potential as toxicity sensors because when their anodes are exposed to toxic substances or the concentration of the target is changed, the voltage generated by certain microorganisms with active electrical activity attached to the anode changes. However, the underlying reasons behind this phenomenon have not been deeply explored. Therefore, the activity of anodic microorganisms during voltage drops in MFCs under formaldehyde (FA) stress was studied. The composition of the microbial community structure with similar active voltage drops under different concentrations of FA was investigated. The results showed that under exposure to high concentrations (169.20 mg/L) of FA, the voltage changes in MFCs could be divided into five stages. With prolonged exposure to FA, the ratio of live/dead bacteria decreased from 4.78 to 0.65. This result indicated that voltage drops may be caused by FA causing cell membrane rupture. Analysis of the microbial community structure under treatments including inoculation and three concentrations of FA revealed that Geobacter and other electrogenic bacteria were effectively enriched in the MFCs. Low-concentration and long-term exposure to FA had a greater impact on microbial communities than high-concentration and short-term exposure. Pseudomonas and Acidovorax were more significantly affected by FA than Flavobacterium and Geobacter. This study explains the reason for the voltage drop of MFCs after exposure to toxic substances from the perspective of the microorganisms.


Parity and Risk of Stroke among Chinese Women: Cross-sectional Evidence from the Dongfeng-Tongji Cohort Study.
Epidemiological studies have investigated the association between parity and the risk of stroke, but the results have been inconsistent. The objective of this study was to assess the association among middle-aged and older Chinese women. We used data from the Dongfeng-Tongji Cohort Study. In total, 14,277 women were included in the analysis. Participants were classified into four groups according to parity. Stroke cases were self-reported during face-to-face interviews. Multivariable logistic regression models were used to investigate the association between parity and the risk of stroke while controlling for potential confounders. The prevalence of stroke among the study subjects was 2.7% (380 of 14,277). In the fully adjusted model, women who had experienced two, three, or four or more live births had 1.24 times (95% CI, 0.85-1.81), 1.97 times (95% CI, 1.30-2.98) and 1.86 times (95% CI, 1.14-3.03), higher risk of stroke, respectively, compared with women who had experienced one live birth. High parity was associated with an increasing risk of stroke in the present study. Further longitudinal studies are needed to confirm the association and to explore the unclear mechanism underlying the link between parity and stroke risk.


Potential use of mangroves as constructed wetland for municipal sewage treatment in Futian, Shenzhen, China.
A pilot-scale mangrove wetland was constructed in Futian, Shenzhen for municipal sewage treatment. Three identical belts (length: 33m, width: 3m, depth: 0.5m) were filled with stone (bottom), gravel and mangrove sand (surface). Seedlings of two native mangrove species (Kandelia candel, Aegiceras corniculatum) and one exotic species (Sonneratia caseolaris) were transplanted to the belts with one species for each belt. The hydraulic loading was 5m(3)d(-1) and hydraulic retention time 3d. High levels of removal of COD, BOD(5), TN, TP and NH(3)-N were obtained. The treatment efficiency of S. caseolaris and A. corniculatum was higher than that of K. candel. Faster plant growth was obtained for S. caseolaris. The substrate in the S. caseolaris belt also showed higher enzyme activities including dehydrogenase, cellulase, phosphatase, urease and beta-glucosidase. The removal rates of organic matter and nutrients were positively correlated with plant growth. The results indicated that mangroves could be used in a constructed wetland for municipal sewage treatment, providing post-treatment to remove coliforms was also included.


Additive manufacturing of metals: a brief review of the characteristic microstructures and properties of steels, Ti-6Al-4V and high-entropy alloys.
We present a brief review of the microstructures and mechanical properties of selected metallic alloys processed by additive manufacturing (AM). Three different alloys, covering a large range of technology readiness levels, are selected to illustrate particular microstructural features developed by AM and clarify the engineering paradigm relating process-microstructure-property. With Ti-6Al-4V the emphasis is placed on the formation of metallurgical defects and microstructures induced by AM and their role on mechanical properties. The effects of the large in-built dislocation density, surface roughness and build atmosphere on mechanical and damage properties are discussed using steels. The impact of rapid solidification inherent to AM on phase selection is highlighted for high-entropy alloys. Using property maps, published mechanical properties of additive manufactured alloys are graphically summarized and compared to conventionally processed counterparts.


Immunity toward H1N1 influenza hemagglutinin of historical and contemporary strains suggests protection and vaccine failure.
Evolution of H1N1 influenza A outbreaks of the past 100 years is interesting and significantly complex and details of H1N1 genetic drift remains unknown. Here we investigated the clinical characteristics and immune cross-reactivity of significant historical H1N1 strains. We infected ferrets with H1N1 strains from 1943, 1947, 1977, 1986, 1999, and 2009 and showed each produced a unique clinical signature. We found significant cross-reactivity between viruses with similar HA sequences. Interestingly, A/FortMonmouth/1/1947 antisera cross-reacted with A/USSR/90/1977 virus, thought to be a 1947 resurfaced virus. Importantly, our immunological data that didn't show cross-reactivity can be extrapolated to failure of past H1N1 influenza vaccines, ie. 1947, 1986 and 2009. Together, our results help to elucidate H1N1 immuno-genetic alterations that occurred in the past 100 years and immune responses caused by H1N1 evolution. This work will facilitate development of future influenza therapeutics and prophylactics such as influenza vaccines.


A comprehensive assessment of regional variation in the impact of head micromovements on functional connectomics.
Functional connectomics is one of the most rapidly expanding areas of neuroimaging research. Yet, concerns remain regarding the use of resting-state fMRI (R-fMRI) to characterize inter-individual variation in the functional connectome. In particular, recent findings that "micro" head movements can introduce artifactual inter-individual and group-related differences in R-fMRI metrics have raised concerns. Here, we first build on prior demonstrations of regional variation in the magnitude of framewise displacements associated with a given head movement, by providing a comprehensive voxel-based examination of the impact of motion on the BOLD signal (i.e., motion-BOLD relationships). Positive motion-BOLD relationships were detected in primary and supplementary motor areas, particularly in low motion datasets. Negative motion-BOLD relationships were most prominent in prefrontal regions, and expanded throughout the brain in high motion datasets (e.g., children). Scrubbing of volumes with FD>0.2 effectively removed negative but not positive correlations; these findings suggest that positive relationships may reflect neural origins of motion while negative relationships are likely to originate from motion artifact. We also examined the ability of motion correction strategies to eliminate artifactual differences related to motion among individuals and between groups for a broad array of voxel-wise R-fMRI metrics. Residual relationships between motion and the examined R-fMRI metrics remained for all correction approaches, underscoring the need to covary motion effects at the group-level. Notably, global signal regression reduced relationships between motion and inter-individual differences in correlation-based R-fMRI metrics; Z-standardization (mean-centering and variance normalization) of subject-level maps for R-fMRI metrics prior to group-level analyses demonstrated similar advantages. Finally, our test-retest (TRT) analyses revealed significant motion effects on TRT reliability for R-fMRI metrics. Generally, motion compromised reliability of R-fMRI metrics, with the exception of those based on frequency characteristics - particularly, amplitude of low frequency fluctuations (ALFF). The implications of our findings for decision-making regarding the assessment and correction of motion are discussed, as are insights into potential differences among volume-based metrics of motion.


Astrocytes adjust the dynamic range of cortical network activity to control modality-specific sensory information processing.
Cortical neuron-astrocyte communication in response to peripheral sensory stimulation occurs in a topographic-, frequency-, and intensity-dependent manner. However, the contribution of this functional interaction to the processing of sensory inputs and consequent behavior remains unclear. We investigate the role of astrocytes in sensory information processing at circuit and behavioral levels by monitoring and manipulating astrocytic activity in vivo. We show that astrocytes control the dynamic range of the cortical network activity, optimizing its responsiveness to incoming sensory inputs. The astrocytic modulation of sensory processing contributes to setting the detection threshold for tactile and thermal behavior responses. The mechanism of such astrocytic control is mediated through modulation of inhibitory transmission to adjust the gain and sensitivity of responding networks. These results uncover a role for astrocytes in maintaining the cortical network activity in an optimal range to control behavior associated with specific sensory modalities.


Disaster preparedness: emergency planning in the NICU.
Hospital emergency management has evolved beyond satisfying regulatory requirements. Although tools and resources have been developed to support hospitals in emergency planning, there appears to be a scarcity of resources to guide hospital departments. To ensure that standards of care are maintained and to minimize the impact on the hospital and/ or a nursing unit, Good Samaritan Hospital has developed a mobile emergency system and an Emergency Operations Plan (EOP) template to assist other nursing units in their planning efforts. This article focuses on the development of emergency bedside backpacks, mobile disaster boxes, disaster documentation and forms go-kits, and guidelines for creating such a plan. The ongoing equipment testing, inventory rotation, staff training, and exercising response protocols are all crucial to test the effectiveness of the program in place. All these activities require a multidisciplinary approach to ensure integration with hospital-wide emergency planning efforts.


Molecular characterization of epidemic multiresistant Staphylococcus haemolyticus isolates.
Fifty-five Staphylococcus haemolyticus specimens isolated from patients and neonatal intensive care unit staff were tested for susceptibility to 12 antimicrobial agents. There were 34 multidrug-resistant isolates which were resistant to oxacillin, ampicillin, cefazolin, cefmetazole, imipenem, and gentamicin. These isolates had a higher frequency of resistance to tobramicin and ofloxacin, and relatively high MICs (2 to 4 micrograms/mL) for vancomycin, although none of the isolates were vancomycin resistant. To investigate hospital-acquired colonization and infection by multiresistant S. haemolyticus, we examined all isolates by pulsed-field gel electrophoresis (PFGE) after SmaI and SstII digestion, and detected an endemic PFGE pattern in multiresistant isolates. The results suggested that local spread of multiresistant S. haemolyticus was hospital acquired, and that the hospital staffs functioned as a reservoir.


AKIP1 promotes tumor progression by cancer-related pathways and predicts prognosis in tongue squamous cell carcinoma.
The current study was designed to explore the effect of A-kinase-interacting protein 1 (AKIP1) on tongue squamous cell carcinoma (TSCC) viability and mobility and to investigate its molecular mechanism. Control overexpression (OE-NC group) and AKIP1 overexpression (OE-AKIP1 group) plasmids were transfected into CAL-27 cells; control knockdown (KD-NC group) and AKIP1 knockdown (KD-AKIP1 group) plasmids were transfected into SCC-9 cells. Cellular viability and mobility were determined, and mRNA sequencing was performed followed by RT-qPCR validation. Immunohistochemistry was utilized to detect AKIP1 expression in tumor and adjacent tissues from 90 TSCC patients. AKIP1 was more highly expressed in human TSCC cell lines compared to human normal lingual epithelial cells. Cell proliferation, migration, and invasion were increased in the OE-AKIP1 group compared to the OE-NC group but decreased in the KD-AKIP1 group compared to the KD-NC group. mRNA sequencing revealed 436 differentially expressed genes; most of the genes were mainly enriched in the mTOR, PI3K-Akt, MAPK, Hippo, and Wnt signaling pathways. These findings were subsequently confirmed by RT-qPCR quantification. In TSCC patients, AKIP1 expression was increased in tumor tissues and related to increased tumor size, lymph node metastasis and poor overall survival. AKIP1 is a therapeutic target that regulates multiple tumor-related pathways in TSCC.


Heavy metal pollution induced by a ferro-nickel smelting plant in Greece.
Seven heavy metals, Co, Cr, Cu, Fe, Mn, Ni and Zn, were measured in marine sediments, plants and invertebrates in the vicinity of a ferro-nickel smelting plant in Greece. The concentrations of metals in the sediment were higher than those found in the average unpolluted Greek coastal sediment. High levels of metals were observed in the gastropod molluscs, particularly Cerithium vulgatum, which concentrated metals more than other invertebrates.


Web-based intervention improves social acknowledgement and disclosure of trauma, leading to a reduction in posttraumatic stress disorder symptoms.
This study aims to investigate the effectiveness of web-based intervention on social acknowledgement and disclosure of trauma and to examine the mediating effect of the improvement in social acknowledgement and disclosure of trauma on the reduction of posttraumatic stress disorder symptoms. A randomized controlled trial was used, with 21 participants in a web-based intervention group and 29 participants in a wait-list control group completing a pre-test and post-test. Results showed that social acknowledgement and disclosure of trauma improved significantly after 1-month intervention, and this improvement mediated the reduction in the posttraumatic stress disorder symptoms. Implications and limitations of this study are discussed.


Comparison of serum and liver hepatitis C virus quasispecies in HCV-related hepatocellular carcinoma.
The hepatitis C virus (HCV) genome consists of quasispecies populations of heterogeneous variants, especially in the hypervariable region. To assess the profiles of viral quasispecies in HCV-related hepatocellular carcinoma, we studied the viral population patterns in serum and liver tissues of 13 HCV-positive patients with hepatocellular carcinoma developed on cirrhotic and non-cirrhotic livers (5 and 8 cases, respectively). HCV genome heterogeneity was analyzed by polymerase chain reaction-mediated single-strand conformation polymorphism analysis, which showed multiple DNA bands representing different hypervariable region sequences. The HCV populations were different between tumorous and nontumorous tissues in 3/5 hepatocellular carcinomas with cirrhosis and in 6/8 without cirrhosis. At least one or more than one common band was detected in both compartments in all but one case. No significant differences in the complexity of HCV quasispecies were found in hepatocellular carcinoma with or without underlying cirrhosis. Comparison of the HCV quasispecies profiles in serum and liver tissues showed a different distribution of HCV variants between these two compartments in 6/7 patients. In four cases, both common and compartmentalized sequences were detected, whereas in two cases, both without cirrhosis, the HCV population in serum was completely different from that found in the liver. These results suggest that the complexity of HCV populations is influenced by the presence of hepatocellular carcinoma rather than by the severity of the underlying chronic liver disease. The different quasispecies patterns found in serum and liver may reflect different biological properties of circulating and intrahepatic HCV particles or the existence of extrahepatic sites of replication.


Effects of acid base disturbance, septic shock, and calcium and phosphorous abnormalities on cardiovascular function.
Systemic acidosis has a negative inotropic effect on myocardial function, which in the intact animal, is counteracted by the activation of the sympathoadrenal system. Although there are extensive animal data in this field, human studies quantifying the influence of systemic acidosis on myocardial function in various disease states is lacking. In patients with hypocalcemia, a single infusion of calcium does not sustain increased calcium levels, and the hemodynamic improvement is only transient. Hemodynamic changes in septic shock are complex, and there are convincing data documenting myocardial dysfunction in sepsis. There is a need for elucidating the biochemical characteristics of the myocardial depressant factor (or factors).


Management of acute agitation in Hong Kong and comparisons with Australasia.
Little is known about the use of sedation drugs for the management of acute agitation in Hong Kong's Accident and Emergency Departments (AEDs) and how it compares with Australasian practice. The aim of this study was to determine drug preferences, clinicians' perceived confidence in management, barriers/gaps in training and perceived usefulness of existing clinical practice guidelines (CPGs) in Hong Kong. A validated questionnaire was used, with case vignettes typical of patients presenting to AEDs with acute agitation. The questionnaire was distributed by hand to all trainees and fellows of the Hong Kong College of Emergency Medicine (HKCEM). Two reminders were sent. Of 483 HKCEM members, 280 (58.0% [95% CI 53.5-62.3]) responded. For monotherapy, 46.8% (95% CI 41.0-52.6) of respondents chose haloperidol to manage the undifferentiated patient, followed by midazolam (33.9%, 95% CI 28.6-39.7) and diazepam (13.9%, 95% CI 10.4-18.5). Most respondents (83.6%, 95% CI 78.8-87.5) would not administer combination therapy. Respondents were confident in managing agitation overall. The lack of local/institutional CPGs (55.7%, 95% CI 49.9-61.4) was perceived as an important barrier. Institutional guidelines were considered the most useful CPGs (66.4%, 95% CI 60.7-71.7). Most respondents (72.9%, 95% CI 67.4-77.7) perceived a HKCEM endorsed CPG would be useful. Haloperidol and benzodiazepines are frequently used as monotherapy for the management of acute agitation in Hong Kong's AEDs. Management in Hong Kong differs from Australasian practice in that combination therapy is less common and clinicians' choice of sedation drugs are less variable overall. Results suggest that future work on CPG development and training regarding the safe use of combination therapy would be well received.


I12: the Joint Engineering, Environment and Processing (JEEP) beamline at Diamond Light Source.
I12 is the Joint Engineering, Environmental and Processing (JEEP) beamline, constructed during Phase II of the Diamond Light Source. I12 is located on a short (5 m) straight section of the Diamond storage ring and uses a 4.2 T superconducting wiggler to provide polychromatic and monochromatic X-rays in the energy range 50-150 keV. The beam energy enables good penetration through large or dense samples, combined with a large beam size (1 mrad horizontally × 0.3 mrad vertically). The beam characteristics permit the study of materials and processes inside environmental chambers without unacceptable attenuation of the beam and without the need to use sample sizes which are atypically small for the process under study. X-ray techniques available to users are radiography, tomography, energy-dispersive diffraction, monochromatic and white-beam two-dimensional diffraction/scattering and small-angle X-ray scattering. Since commencing operations in November 2009, I12 has established a broad user community in materials science and processing, chemical processing, biomedical engineering, civil engineering, environmental science, palaeontology and physics.


Fetal sex prediction measuring yolk sac size and yolk sac-fetal pole distance in the first trimester via ultrasound screening.
The aim of the study was to determine the accuracy of fetal sex definition by measuring the yolk sac size and the yolk sac-fetal pole distance in the first trimester via ultrasound (US) screening. We enrolled 92 gestational women in this prospective study. In the first trimester of US examination, the gestational sac, the gestational sac-fetal pole distance, and the yolk sac size were measured. When the fetal pole was in the longitudinal position, the distance between the fetal pole and the yolk sac was measured in millimeters (mm). The crown-rump length (CRL) was measured in the same position. US examination at 22 weeks was performed to determine whether the fetal gender was male or female. The genders of the fetuses were recorded. The sexes were also confirmed and recorded after birth. Fifty-five (59.8%) of the newborns were found to be male, while 37 (40.2%) of them were found to be female. We compared the yolk sac size and the yolk sac-fetal pole distance in both the female and male groups. We found that the genders showed a significant difference in terms of yolk sac-fetal pole distance, whereas the yolk sac size showed no significant difference between the genders. The optimal cutoff value of the yolk sac-fetal pole distance was 1.80 mm, with 70% sensitivity and 67% specificity for female gender prediction. The distance also showed an independent association with gender prediction in the first trimester. Yolk sac-fetal pole distance may have the potential to predict gender in the first trimester of pregnancy.


Crystal structure of the OrfX1-OrfX3 complex from the PMP1 neurotoxin gene cluster.
Paraclostridial mosquitocidal protein 1 (PMP1) is a member of the clostridial neurotoxin (CNT) family, which includes botulinum and tetanus neurotoxins. PMP1 has unique selectivity for anopheline mosquitos and is the only known member of the family that targets insects. PMP1 is encoded in an orfX gene cluster, which in addition to the toxin, consists of OrfX1, OrfX2, OrfX3, P47 and NTNH, which have been shown to aid in PMP1 toxicity. We here show that OrfX1 and OrfX3 form a complex and present its structure at 2.7 Å. The OrfX1-OrfX3 complex mimics the structure of full-length OrfX2 and belongs to the lipid-binding TULIP protein superfamily. With this report, the structures of all proteins encoded in the orfX gene cluster of CNTs are now determined.


Growth arrest failure, G1 restriction point override, and S phase death of sensory precursor cells in the absence of neurotrophin-3.
More than half of the dorsal root ganglion (DRG) neurons are lost by excessive cell death coinciding with precursor proliferation and cell cycle exit in neurotrophin-3 null mutant (NT-3-/-) mice. We find that in the absence of NT-3, sensory precursor cells fail to arrest the cell cycle, override the G1 phase restriction point, and die by apoptosis in S phase, which can be prevented in vivo by a cell cycle blocker. Uncoordinated cell cycle reentry is preceded by a failure of nuclear N-myc downregulation and is paralleled by the activation of the full repertoire of G1 and S phase cell cycle proteins required for cell cycle entry. Our results provide evidence for novel activity of neurotrophins in cell cycle control and point toward an N-myc sensitization to cell death in the nervous system that is under the control of NT-3.


Integrity of a heterochromatic domain ensured by its boundary elements.
In fission yeast, the inverted repeats IR-L and IR-R function as boundary elements at the edges of a 20-kb silent heterochromatic domain where nucleosomes are methylated at histone H3K9. Each repeat contains a series of B-box motifs physically associated with the architectural TFIIIC complex and with other factors including the replication regulator Sap1 and the Rix1 complex (RIXC). We demonstrate here the activity of these repeats in heterochromatin formation and maintenance. Deletion of the entire IR-R repeat or, to a lesser degree, deletion of just the B boxes impaired the de novo establishment of the heterochromatic domain. Nucleation proceeded normally at the RNA interference (RNAi)-dependent element cenH but subsequent propagation to the rest of the region occurred at reduced rates in the mutants. Once established, heterochromatin was unstable in the mutants. These defects resulted in bistable populations of cells occupying alternate "on" and "off" epigenetic states. Deleting IR-L in combination with IR-R synergistically tipped the balance toward the derepressed state, revealing a concerted action of the two boundaries at a distance. The nuclear rim protein Amo1 has been proposed to tether the mating-type region and its boundaries to the nuclear envelope, where Amo1 mutants displayed milder phenotypes than boundary mutants. Thus, the boundaries might facilitate heterochromatin propagation and maintenance in ways other than just through Amo1, perhaps by constraining a looped domain through pairing.


His92Ala mutation in ribonuclease T1 induces segmental flexibility. An X-ray study.
In the genetically mutated ribonuclease T1 His92Ala (RNase T1 His92Ala), deletion of the active site His92 imidazole leads to an inactive enzyme. Attempts to crystallize RNase T1 His92Ala under conditions used for wild-type enzyme failed, and a modified protocol produced two crystal forms, one obtained with polyethylene glycol (PEG), and the other with phosphate as precipitants. Space groups are identical to wild-type RNase T1, P2(1)2(1)2(1), but unit cell dimensions differ significantly, associated with different molecular packings in the crystals; they are a = 31.04 A, b = 62.31 A, c = 43.70 A for PEG-derived crystals and a = 32.76 A, b = 55.13 A, c = 43.29 A for phosphate-derived crystals, compared to a = 48.73 A, b = 46.39 A, c = 41.10 A for uncomplexed wild-type RNase T1. The crystal structures were solved by molecular replacement and refined by stereochemically restrained least-squares methods based on Fo greater than or equal to sigma (Fo) of 3712 reflections in the resolution range 10 to 2.2 A (R = 15.8%) for the PEG-derived crystal and based on Fo greater than or equal to sigma (Fo) of 6258 reflections in the resolution range 10 to 1.8 A (R = 14.8%) for the phosphate-derived crystal. The His92Ala mutation deletes the hydrogen bond His92N epsilon H ... O Asn99 of wild-type RNase T1, thereby inducing structural flexibility and conformational changes in the loop 91 to 101 which is located at the periphery of the globular enzyme. This loop is stabilized in the wild-type protein by two beta-turns of which only one is retained in the crystals obtained with PEG. In the crystals grown with phosphate as precipitant, both beta-turns are deleted and the segment Gly94-Ala95-Ser96-Gly97 is so disordered that it is not seen at all. In addition, the geometry of the guanine binding site in both mutant studies is different from "empty" wild-type RNase T1 but similar to that found in complexes with guanosine derivatives: the Glu46 side-chain carboxylate hydrogen bonds to Tyr42 O eta; water molecules that are present in the guanine binding site of "empty" wild-type RNase T1 are displaced; the Asn43-Asn44 peptide is flipped such that phi/psi-angles of Asn44 are in alpha L-conformation (that is observed in wild-type enzyme when guanine is bound).(ABSTRACT TRUNCATED AT 400 WORDS)


[Measurement of success of treatment for chronic back pain. Can general effects in an entire sample of patients with chronic back pain be transferred to specific effects in individuals?].
The multimodal therapy has proved to be useful for the treatment of patients with chronic back pain. The aim of this methodical study was to test whether specific effects for the individual can be inferred from general effects of the entire sample, in order to define criteria for the success of treatment. These could be helpful when deciding to which patients the multimodal therapy can best be applied. A total of 408 patients with chronic back pain took part in a 3-week multimodal therapy program in a day hospital. The patients were questioned regarding parameters such as pain intensity, depression, functional back capacity and quality of life before therapy as well as 6 months after therapy. On average, a significant improvement of all parameters could be shown after 6 months. However, scatterplots and correlations showed that general results of the entire sample should not be transferred to the individual patient Taking into account principles of classical test theory this study showed that sufficient systematic coherence of the data must be proven before changes can be calculated. As long as neither an adequate systematic nor useful criteria can be found, there seems to be no reason to exclude any patient from multimodal therapy using pre-selection criteria.


Vitamin C supplementation in normal subjects reduces constitutive ICAM-1 expression.
Regulation of monocyte adhesion molecule gene expression is via redox sensitive transcription factors. We have investigated whether dietary antioxidant supplementation with vitamin C (250 mg/day) can modulate monocyte ICAM-1 expression in healthy male subjects with low plasma vitamin C at baseline. In a randomised, double-blind, crossover study, monocyte ICAM-1 mRNA was analysed using quantitative reverse transcriptase PCR. Protein was determined by flow cytometry (monocytes) and ELISA (plasma). Monocyte numbers were unaltered by supplementation. Subjects with low plasma vitamin C (<50 microM) prior to supplementation expressed higher levels of monocyte ICAM-1mRNA, and showed a significant (50%) reduction in ICAM-1mRNA expression after 6 weeks of 250 mg/day vitamin C supplementation (p<0.05). This was paralleled by a reduction in sICAM-1 (p<0.05). For the first time, these results show that dietary vitamin C can modulate monocyte ICAM-1 gene expression in vivo, where regulation of gene expression represents a novel mechanism for benefit from dietary antioxidants.


Alpha-adrenoreceptor activation modulates swimming via glycinergic and GABAergic inhibitory pathways in Xenopus laevis tadpoles.
This study focuses upon the network pathways underlying the adrenoreceptor-mediated modulation of fictive swimming in the immobilized Xenopus laevis tadpole. As shown recently, noradrenaline (NA) increases cycle periods while simultaneously reducing the rostrocaudal delay in head-to-tail firing and the duration of swimming episodes. Furthermore, both swimming frequency and duration are reduced by selective pharmacological activation of alpha1- and/or alpha2-adrenoreceptors, while alpha1-receptor activation also reduces rostrocaudal delays. We show that NA could still modulate aspects of swimming after blocking either glycine or GABA(A) receptors with strychnine and bicuculline, respectively. Furthermore, after prior application of NA, strychnine could counteract noradrenergic effects on cycle periods and rostrocaudal delays, while bicuculline could counteract effects on cycle periods, suggesting that these two fast inhibitory pathways are both involved in the NA modulation of swimming. In addition, blocking glycine receptors reduced the effects of alpha1-receptors on cycle periods and delays, while blocking GABA(A) receptors had no effect. Blocking either glycine or GABA(A) receptors, however, lessened the reduction in swimming frequency by alpha2-receptors. In addition, pre-application of bicuculline prevented a reduction in episode durations by NA, alpha1- and alpha2-receptors. Our findings suggest that the noradrenergic modulation of Xenopus swimming is mediated via alpha-adrenoreceptors interacting with both glycinergic and GABAergic inhibitory pathways. Both alpha1- and alpha2-receptor activation influences the GABAergic pathway controlling the duration of swimming episodes and is involved in the glycinergic modulation of the swimming rhythm and its longitudinal co-ordination, with alpha2-receptors additionally affecting swimming frequency through GABAergic pathways.


Discrimination of systolic blood pressure.
The ability of humans to discriminate systolic blood pressure (BP) was investigated in two experiments. In Experiment 1, 14 normal subjects were asked to make estimates of their systolic BP while performing both BP-elevating and BP-lowering tasks. They were given intermittent feedback throughout all 10 45-min sessions. Results indicated significant correlations and small absolute differences between estimated and measured BP for all subjects in almost all sessions. Experiment 2, undertaken 6 months after Experiment 1, assessed whether estimation accuracy by subjects who had available both external and interoceptive cues surpassed that of subjects which access to external cues only. Three subjects from the original group who showed consistently high motivation, and who improved in accuracy across the 10 sessions in the previous experiment, made estimates of BP while performing novel tasks with no feedback. Correlations between estimated and measured BP remained high for 2 of the 3. These results were compared with the accuracy of control subjects (3 for each experimental subject) who were asked to estimate experimental subjects' BP using only the cognitive information available to the experimental subjects. Control subjects also had high correlations between their estimates and the experimental subjects' measured BP but at lower levels than two experimental subjects. These findings are discussed in relation to subjects' possible use of interoceptive information.


Mediation of human NK-activity by components in extracts of Viscum album.
Viscum album extracts (Iscador) were investigated for their potency to influence NK cytotoxicity in vitro. In vitro short term cytotoxicity assays (4 h) with human peripheral mononuclear cells (PMNC) and human K 562 tumor cells showed a drastic enhancement of NK cytotoxicity in the presence of V. album extracts. The presence of the V. album components during tumor cell lysis was essential since preincubation of PMNC with V. album extract followed by thorough washing did not lead to enhancement of NK cytotoxicity. One responding effector cell was identified as a member of the large granular lymphocyte (LGL) family carrying both Leu 7 and Leu 11 surface markers. Furthermore, monocytes depleted of LGL, but not differentiated macrophages, showed a weak enhancement of their cytolytic activity in the presence of V. album extract. Fractionation of V. album extracts revealed two active fractions one (C1) with about 3-4000 D and the other (C2) less than 1000 D. Both components enhanced NK cytotoxicity of LGL (Leu 7+, Leu 11+) as well as of monocytes showing enhancing effects also against moderately NK-sensitive tumor cell lines.


Analysis of the effect of breast magnetic resonance imaging on the outcome in women undergoing breast conservation surgery with radiation therapy.
It remains uncertain whether MRI identification of additional foci of disease leads to improved outcome. We undertook a study to evaluate the influence of breast MRI on early and long-term outcome. Among 1,271 patients undergone breast cancer surgery between January 2005 and December 2006, 785 patients were attempted for BCS. Operative approach and radiologic findings were compared according to MRI use. We reselected 615 patients with unilateral early-stage breast cancer treated with BCS including RT. We compared the histopathologic characteristics and outcomes according to MRI use. In patients attempted for BCS (n = 785), re-excision rates were not significantly different according to MRI use (P = 1.000). Conversion to mastectomy or bilateral cancer surgery were higher in MRI group (P = 0.002). The IBTR rate was higher in the non-MRI group (P = 0.020). Difference in contralateral cancer rate and total recurrence rates failed to reach statistical significance (P = 0.168, 0.383, respectively). Multivariate study after adjustment showed no difference in recurrence rates and IBTR rates between the two groups (hazard ratios 1.34, 6.37 Ps = 0.385, 0.076). Use of MRI in patients with early-stage breast cancer did not result in improvement of a patient's outcome.


Demonstration of intrinsic efflux activity of Escherichia coli K-12 AG100 by an automated ethidium bromide method.
Demonstration of efflux of ethidium bromide (EtBr) has been made for over 30 bacterial species, usually by showing enhanced efflux in multidrug-resistant strains that was then abolished by inactivating efflux pumps. Here we present a relatively simple automated method that employs EtBr as an efflux pump substrate for the demonstration of intrinsic efflux activity in Escherichia coli K-12 AG100. The method uses the Rotor-Gene 3000 instrument for real-time fluorometric measurement of EtBr accumulation under conditions that limit energy (absence of glucose, low temperature) and of EtBr extrusion under optimum conditions. The method can be used for screening compound libraries for efflux inhibiting capacity.


Enzyme cytochemical localization of sarcosine oxidase activity in the liver and kidney of several mammals.
We investigated the enzyme cytochemical localization of sarcosine oxidase (SOX) in the liver and kidney of several mammals using a cerium technique. First we measured the enzyme activities in the liver and kidney of several mammals and in several organs of mice. The highest activity was found in the Chinese hamster, followed by the mouse. Therefore, we used hamster and mouse tissues for enzyme cytochemistry. The liver and kidneys were fixed by perfusion with various concentrations of glutaraldehyde for 10 min. Tissue slices were incubated in reaction medium consisting of 50 mM TRIS-maleate buffer (pH 7.8), 9 mM sodium azide, 9.8 mM sarcosine, 25 microM FAD, 2 mM cerium chloride, 0.002% saponin, and 0.003% Triton X-100 for 0.5-8 h at 37 degrees C. Optimum staining reaction was obtained in tissues fixed with 0.2% glutaraldehyde, followed by incubation for 2-4 h. Electron-dense reaction products were present exclusively in peroxisomes. Within the peroxisomes strong reactions were observed in the matrix subjacent to the limiting membrane decreasing toward the center. The staining reaction was completely inhibited by 2 mM N-bromosuccinimide. These results indicated that SOX is a peroxisomal enzyme and that the enzyme might be associated with the peroxisomal membrane.


Solvent-free cross-dehydrogenative coupling reactions under high speed ball-milling conditions applied to the synthesis of functionalized tetrahydroisoquinolines.
Solvent-free reaction using a high-speed ball milling technique has been first applied to cross-dehydrogenative coupling (CDC) reactions between tetrahydroisoquinolines and three types of pronucleophiles such as nitroalkanes, alkynes, and indoles. All coupling products were obtained in good yields at short reaction times (no more than 40 min). When alkynes and indoles were used as pronucleophile, the reactions can be catalyzed efficiently by recoverable copper balls without any additional metal catalyst.


Derivatisation in gas chromatographic determination of acidic herbicides in aqueous environmental samples.
This paper reviews derivatisation processes applied in chromatographic determination of acidic herbicides (with carboxyl and phenol groups), mainly in aqueous environmental samples. The discussion focuses on the basic derivatisation reactions used to convert herbicides to derivatives to make them analysable by gas chromatography, and possibly to reduce detection limits and/or increase extraction recovery from aqueous samples. The reactions are transesterification, esterification, silylation, alkylation, and extractive and pyrolytic alkylation. The reagents used to conduct the reactions are numerous. Diazomethane is a very efficient methylation reagent but explosive and toxic. Methyl iodide also ensures rapid and efficient methylation. Benzyl bromide can be used directly in water but derivatisation yield is low and reproducibility is poor. Butyl chloroformate and dimethyl sulfite can also be used for derivatisation in water, and acetic anhydride can be used for in-situ derivatisation of phenolic herbicides. For increasing selectivity of GC detection pentafluorobenzyl bromide (for ECD) and (2-cyanethyl)dimethyl(diethylamino)silane (for NPD) have been applied. Very characteristic ions are produced in mass spectra if silyl groups are introduced, e.g. by using bis(trimethylsilyl)trifluoroacetamide. Tetramethylammonium, trimethylphenylammonium, tetraalkylammonium, and trimethylsulfonium hydroxides and salts can by used for derivatisation at elevated temperature in the GC injection port. Extractive alkylation is relatively efficient if tetraalkylammonium salts with long chain alkyl groups are used.


Periocular skin warming promotes body heat loss and sleep onset: a randomized placebo-controlled study.
Periocular skin warming was reported to have favorable effects on subjective and objective sleep quality. We hypothesized that enhancing body heat loss by periocular skin warming would reduce sleep onset and improve sleep quality. Eighteen healthy volunteers were asked to maintain wakefulness with their eyes closed for 60 min after applying either a warming or sham eye mask, followed by a 60-min sleep period. Compared to the sham, periocular warming increased the distal skin temperature and distal-proximal skin temperature gradient only during the 30-min thermal manipulation period. In the subsequent sleep period, periocular warming facilitated sleep onset, increased stage 2 sleep and electroencephalographic delta activity during the first half of the sleep period relative to the sham. These results suggest that periocular skin warming may accelerate and deepen sleep by enhancing physiological heat loss via the distal skin, mimicking physiological conditions preceding habitual sleep.


Cultures of glial cells release purines under field electrical stimulation: the possible ionic mechanisms.
Dissociated primary cultures of glial cells released a remarkable amount of purines, at rest and during field electrical stimulation. The HPLC identification of labelled compounds derived from 3H-Adenosine (3H-Ado) (employed to preload the cultures) indicated that nucleotides and nucleosides were represented in the superfusate in equivalent proportions (43.86% and 56.14% respectively). Very much higher amounts of unlabelled purines prevalently constituted by nucleotides compounds (91.10%) were also released and detectable in the superfusate. In all the experimental conditions their evoked release did not result frequency-dependent. Since: a linear increase related to the stimulation frequencies was found for the released labelled compounds; no labelled purines were assayed in 5 x 10-5M Dipyridamole-treated cultures; any significant presence of labelled nucleotides, inosine and hypoxantine was not found in cultures simultaneously treated with 1 x 10-5M 2'-deoxycoformycin and 1 x 10-4M 1-(-5-isoquinolinsulfonyl)-2-methylpiperizine (H7) (3H-Ado amounts resulted more than doubled in these experimental conditions); labelled compounds have been assumed as tracers of a glial purine rate whose release can be connected to electrically-evoked action potentials. Purine outflow from glial cells is not sodium dependent, in fact TTX (5 x 10-7M) did not affect their basal or electrically-evoked release. A remarkable calcium-dependence was also evidentiated by the 1 x 10-4M Verapamil-induced inhibition of basal and evoked release. TEA (1 x 10-2M), a specific inhibitor of potassium efflux throughout calcium-mediated specific channels, strongly reduced the evoked purine outflow and any additive effect of its was not detectable when administered simultaneously to the calcium antagonist. These findings indicate that the frequency-dependent purine release from cultured glial cells is linked to ionic mechanisms, which calcium and potassium are mainly involved in.


"Is magnesium depletion the reason for ototoxicity caused by aminoglycosides?".
Aminoglycoside antibiotic drugs may cause ototoxicity and nephrotoxicity. Our hypothesis postulates that aminoglycosides cause ototoxicity by a mechanism of magnesium depletion in the hair cells of the cochlea. The same mechanism maybe responsible for nephrotoxicity caused by aminoglycosides.


Global characterization of macrophage polarization mechanisms and identification of M2-type polarization inhibitors.
Macrophages undergoing M1- versus M2-type polarization differ significantly in their cell metabolism and cellular functions. Here, global quantitative time-course proteomics and phosphoproteomics paired with transcriptomics provide a comprehensive characterization of temporal changes in cell metabolism, cellular functions, and signaling pathways that occur during the induction phase of M1- versus M2-type polarization. Significant differences in, especially, metabolic pathways are observed, including changes in glucose metabolism, glycosaminoglycan metabolism, and retinoic acid signaling. Kinase-enrichment analysis shows activation patterns of specific kinases that are distinct in M1- versus M2-type polarization. M2-type polarization inhibitor drug screens identify drugs that selectively block M2- but not M1-type polarization, including mitogen-activated protein kinase kinase (MEK) and histone deacetylase (HDAC) inhibitors. These datasets provide a comprehensive resource to identify specific signaling and metabolic pathways that are critical for macrophage polarization. In a proof-of-principle approach, we use these datasets to show that MEK signaling is required for M2-type polarization by promoting peroxisome proliferator-activated receptor-γ (PPARγ)-induced retinoic acid signaling.


Identification of potential type III secretion proteins via heterologous expression of Vibrio parahaemolyticus DNA.
We employed a heterologous secretion assay to identify proteins potentially secreted by type III secretion systems (T3SSs) in Vibrio parahaemolyticus. N-terminal sequences from 32 proteins within T3SS genomic islands and seven proteins from elsewhere in the chromosome included proteins that were recognized for export by the Yersinia enterocolitica flagellar T3SS.


Revisiting the octopicolid copepods (Octopicolidae: Octopicola Humes, 1957): comparative morphology and an updated key to species.
A review of the present state of knowledge on the octopicolid copepods (Octopicolidae: Octopicola Humes, 1957) is presented. Characteristic morphological features are illustrated with scanning electron micrographs of Octopicola superba superba Humes, 1957. Comparative morphology analysis led to the conclusion that there is sufficient evidence to justify raising the two subspecies of O. superba to full species rank. A new identification key for the four species of Octopicola Humes, 1957, i.e. O. superba Humes, 1957, O. antillensis Stock, Humes & Gooding, 1963, O. stocki Humes, 1963 and O. regalis Humes, 1974, is proposed after evaluation of the morphological characters which vary more markedly between them. Among other characters, these species differ in the ornamentation of the third antennal segment, maxilla and male maxilliped. They are further distinguished by a combination of several character states concerning the fifth pedigerous somite.


Application of cluster analysis to the geochemistry zonation of the estuary waters in the Tinto and Odiel rivers (Huelva, Spain).
The combination of acid water from mines, industrial effluents and sea water plays a determining role in the evolutionary process of the chemical makeup of the water in the estuary of the Tinto and Odiel rivers. This estuary is in the southwest of the Iberian Peninsula and is one of the estuarine systems on the northwest coast of the Gulf of Cádiz. From the statistical treatment of data obtained by analyzing samples of water from this system, which is affected by industrial and mining pollution processes, we can see how the sampling points studied form two large groups depending on whether they receive tidal or fluvial influences. Fluvial input contributes acid water with high concentrations of heavy metal, whereas industrial effluents are responsible for the presence of phosphates, silica and other nutrients. The estuarine system of the Tinto and Odiel Rivers can be divided into three areas--the Tinto estuary, the Odiel estuary and the area of confluence--based on the physical--chemical characteristics of the water.


Identification of a putative membrane receptor for the bioactive phospholipid, lysophosphatidic acid.
Lysophosphatidic acid (LPA) is a naturally occurring phospholipid with hormone- and growth factor-like activities. Exogenous LPA stimulates GTP-dependent phosphoinositide hydrolysis and inhibits adenylate cyclase in its target cells, but the site of action of LPA is unknown. We now report the identification by photoaffinity labeling of a putative LPA membrane receptor in various LPA-responsive cell types. A 32P-labeled LPA analogue containing a photoreactive fatty acid, [32P]diazirine-LPA, labels a membrane protein of apparent molecular mass of 38-40 kDa in various cell types, including neuronal cells, brain homogenates, carcinoma cells, leukemic cells and normal fibroblasts. Labeling of the 38-40 kDa protein is competitively inhibited by unlabeled 1-oleoyl-LPA (IC50 approximately 10 nM), but not by other phospholipids. Specific labeling is not detected in rat liver membranes or in human neutrophils, which are physiologically unresponsive to LPA. Suramin, an inhibitor of both early and late events in the action of LPA, completely inhibits the binding of photoreactive LPA. We suggest that the 38-40 kDa protein represents a specific LPA cell surface receptor mediating at least part of the multiple cellular responses to LPA.


Fecal microbiota transplantation improves chicken growth performance by balancing jejunal Th17/Treg cells.
Intestinal inflammation has become a threatening concern in chicken production worldwide and is closely associated with Th17/Treg cell imbalance. Several studies described that gut microbiota is significantly implicated in chicken growth by modulating intestinal immune homeostasis and immune cell differentiation. Whether reshaping gut microbiota by fecal microbiota transplantation (FMT) could improve chicken growth by balancing Th17/Treg cells is an interesting question. Here, the chickens with significantly different body weight from three different breeds (Turpan cockfighting × White Leghorn chickens, white feather chickens, and yellow feather chickens) were used to compare Th17 and Treg cells. qPCR and IHC staining results indicated that Th17 cell-associated transcriptional factors Stat3 and rorγt and cytokines IL-6, IL-17A, and IL-21 were significantly (P < 0.05) higher in the jejunum of low body weight chickens, while Treg cell-associated transcriptional factor foxp3 and cytokines TGF-β and IL-10 were significantly (P < 0.05) lower in the jejunum of low body weight chickens, indicating imbalanced Th17/Treg cells were closely related to chicken growth performance. Transferring fecal microbiota from the healthy donor with better growth performance and abundant Lactobacillus in feces to 1-day-old chicks markedly increased growth performance (P < 0.001), significantly decreased Th17 cell-associated transcriptional factors and cytokines, and increased Treg cell-associated transcriptional factors and cytokines in the jejunum (P < 0.05). Furthermore, FMT increased the abundance of Lactobacillus (FMT vs Con; 84.98% vs 66.94%). Besides, the metabolites of tryptophan including serotonin, indole, and 5-methoxyindoleacetate were increased as well, which activated their receptor aryl-hydrocarbon-receptor (AhR) and expressed more CYP1A2 and IL-22 to maintain Th17/Treg cell balance and immune homeostasis. These findings suggested that imbalanced Th17/Treg cells decreased chicken growth performance, while FMT-reshaped gut microbiota, i.e., higher Lactobacilli, increased chicken growth performance by balancing Th17/Treg cells. Video Abstract.


Clinical pharmacokinetics and endocrine disorders. Therapeutic implications.
Endocrine disorders are common and produce widespread changes in cellular and organ function. Alterations in the sensitivity of patients with thyroid disorders to digoxin, anticoagulants and sedatives have been recognised for many years. Many of the recently documented kinetic alterations in endocrine patients are explicable on the basis of disease-induced changes in hepatic drug metabolism, protein binding and renal function. In hyperthyroidism the rate of absorption of paracetamol, propranolol and oxazepam is increased due to increased gastrointestinal motility. The volume of distribution of propranolol and digoxin is increased and there is decreased binding of both basic and acidic drugs as a consequence of alterations in alpha 1-acid glycoprotein and albumin concentration. The rate of glucuronidation of paracetamol and oxazepam is increased in hyperthyroidism. While oxidative metabolism of antipyrine, propranolol, metoprolol and theophylline is enhanced, the clearance of a number of other agents, including diazepam, warfarin, antithyroid drugs and phenytoin, is unaltered. The systemic clearance of propranolol is enhanced as a consequence of a 50% increase in liver blood flow. The rate of elimination of a number of endogenous substances, including cortisol, thyroid hormones and insulin, also appear to be enhanced. Hyperthyroidism has a variable effect on renal function, with a possible increase in digoxin elimination, but no effect on the clearance of renally excreted beta-blockers, atenolol, sotalol and nadolol. These kinetic changes suggest that individualization and higher than normal dosage of propranolol is necessary to control hyperthyroidism, and in thyrotoxic atrial fibrillation higher doses of digoxin or additional therapy with beta-blockers, or verapamil, may be indicated. The increased sensitivity of thyrotoxic patients to warfarin suggest care with dosage and frequent monitoring of response are warranted. Less information is available concerning hypothyroidism, but there is a general trend for decreased absorption of paracetamol and propranolol. In addition, the volume of distribution of digoxin is reduced, as is renal clearance. Limited studies suggest no alteration in the glucuronidation of oxazepam, but antipyrine clearance appears to be reduced. Steady-state propranolol concentrations are elevated in hypothyroidism and there appears to be a decreased metabolism of thyroid hormones and cortisol. Preliminary information suggests the binding of propranolol is increased. Thus, in the treatment of hypothyroid patients, a lower dosage of propranolol may be required.(ABSTRACT TRUNCATED AT 400 WORDS)


Measurement of tobramycin by reversed-phase high-performance liquid chromatography with mass spectrometry detection.
Analysis of tobramycin faces challenges owing to its significant basicity, hydrophilicity and lack of a UV absorbing chromophore. Chromatographic methods, coupled with derivatization to introduce chromophores for tobramycin analysis, were extensively studied. A direct reversed-phase HPLC method for tobramycin analysis has not been reported. Here, we would like to report a simple LC/MS method for quantitative analysis of tobramycin in pharmaceutical formulations. Reversed-phase HPLC analysis of tobramycin was achieved using a pH stable C18 column with basic (pH 11) aqueous mobile phase (ammonium hydroxide buffer), while direct detection was carried out employing a single quadruple mass detector in negative mode via electrospray ionization. This unique separation-detection combination provided simple and specific determination of tobramycin. This method was found to be linear at a tobramycin concentration range of 0.2-0.8 mg/mL with a correlation coefficient value of 0.999. The quantitation limit and detection limit were calculated as 0.210 and 0.063 microg/mL, respectively, with 99.994% confidence. This method was successfully applied to measure tobramycin content in matrices containing tobramycin and other pharmaceutical formulation ingredients. Recoveries of 101.8, 97.8 and 106.7% were obtained for tobramycin spiked in the pharmaceutical formulation at concentrations of 1.68, 1.0 and 0.35 mg/mL, respectively. The relative standard deviations for six injections of spiked samples ranged from 0.2 to 3.2%, indicating good method repeatability.


Thromboembolic disease after combined anterior/posterior reconstruction for adult spinal deformity: a prospective cohort study using magnetic resonance venography.
Thromboembolic disease (TED) after anterior/posterior spinal reconstructions was prospectively evaluated in 66 consecutive patients. Determine the incidence of TED after anterior/posterior spinal reconstruction. Few studies have examined the rate of TED after potentially high-risk combined anterior/posterior reconstructions. Magnetic resonance venography (MRV) is more effective at detecting pelvic deep venous thrombosis (DVT) than conventional screening, but has not been used in these patients. We undertook a prospective cohort study of 66 consecutive adult patients (mean, 52.7 +/- 9.6 years) undergoing combined anterior/posterior spinal reconstructions for spinal deformity. All patients received only mechanical DVT prophylaxis. After surgery, MRV and bilateral lower extremity Doppler ultrasounds were obtained to screen for DVT, and contrast-enhanced spiral computed tomography scans were obtained for clinical suspicion of pulmonary embolism (PE). The total incidence of postoperative TED was 13.6% (9 patients). The overall rate of DVT was 9.1% (6 patients), one-third occurring in the pelvis. PE developed in 7.6% (5 patients). In 2 patients, PE developed less than 48 hours after positive dopplers. In 3 patients, PE developed despite negative screening studies. Right-sided thoracoabdominal approaches were associated with an increased risk of developing DVT (P = 0.03, Odds Ratio 9.8), PE (P = 0.01, Odds Ratio 20), and TED (P = 0.004, Odds Ratio 12). We report a high rate of TED after extensive anterior/posterior spinal reconstructions, for which a right-sided thoracoabdominal approach is an independent risk factor and screening ineffective at preventing PE. These patients should be considered at high risk for postoperative TED.


Arterial dilations in response to calcitonin gene-related peptide involve activation of K+ channels.
Calcitonin gene-related peptide (CGRP) is a 37-amino-acid peptide produced by alternative processing of messenger RNA from the calcitonin gene. CGRP is one of the most potent vasodilators known. It occurs in and is released from perivascular nerves and has been detected in the blood stream, suggesting that it is important in the control of blood flow. The mechanism by which it dilates arteries is not known. Here, we report that arterial dilations in response to CGRP are partially reversed by blockers of the ATP-sensitive potassium channel (K(ATP)), glibenclamide and barium. We also show that CGRP hyperpolarizes arterial smooth muscle and that blockers of K(ATP) channels reverse this hyperpolarization. Finally, we show that CGRP opens single K+ channels in patches on single smooth muscle cells from the same arteries. We propose that activation of K(ATP) channels underlies a substantial part of the relaxation produced by CGRP.


Sexually transmitted diseases.
Sexually transmitted infections (STIs), also referred to as sexually transmitted diseases, remain a growing worldwide problem and public health issue. This article covers the epidemiology of STIs, the history and physical findings, screening guidelines, and the general plan to combat STIs. Prevention is discussed using the latest information from the Centers for Disease Control and Prevention and other references. Infections discussed from the standpoint of cause, epidemiology, risk factors, clinical disease, diagnosis, and treatment include gonorrhea, Chlamydia trachomatis, Trichomonas vaginalis, syphilis, chancroid, Herpes simplex, lymphogranuloma venereum, granuloma inguinale, Herpes papilloma virus, Molluscum contagiosum, and pubic lice.


The effects of systemic and local microinjection into the central nervous system of the selective serotonin 5-HT6 receptor agonist WAY-208466 on sleep and wakefulness in the rat.
The effects of WAY-208466, a selective 5-HT6 receptor agonist on spontaneous sleep were studied in adult rats implanted for chronic sleep recordings. Systemic administration of WAY-208466 during the light phase of the light-dark cycle significantly increased wakefulness (W) and reduced slow wave sleep (SWS), REM sleep (REMS) and the number of REMS periods. Pretreatment with the selective 5-HT6 receptor antagonist RO-399885 prevented the effects of the 5-HT6 receptor agonist on W, SWS and REMS. Direct infusion of WAY-208466 into the dorsal raphe nucleus, locus coeruleus, basal forebrain (horizontal limb of the diagonal band of Broca) or laterodorsal tegmental nucleus specifically decreased REMS without significantly altering W or SWS. In all instances the REMS suppression was dependent upon the reduction of REMS periods. The finding that WAY-208466 increases extracellular γ-aminobutyric acid (GABA) levels in the rat frontal cortex tends to suggest that the neurotransmitter could be involved in the 5-HT6 receptor agonist-induced disruption of the sleep-wake cycle. However, further studies are needed to resolve this issue.


Hemodynamic alternations following stent deployment and post-dilation in a heavily calcified coronary artery: In silico and ex-vivo approaches.
In this work, hemodynamic alterations in a patient-specific, heavily calcified coronary artery following stent deployment and post-dilations are quantified using in silico and ex-vivo approaches. Three-dimensional artery models were reconstructed from OCT images. Stent deployment and post-dilation with various inflation pressures were performed through both the finite element method (FEM) and ex vivo experiments. Results from FEM agreed very well with the ex-vivo measurements, interms of lumen areas, stent underexpansion, and strut malapposition. In addition, computational fluid dynamics (CFD) simulations were performed to delineate the hemodynamic alterations after stent deployment and post-dilations. A pressure time history at the inlet and a lumped parameter model (LPM) at the outlet were adopted to mimic the aortic pressure and the distal arterial tree, respectively. The pressure drop across the lesion, pertaining to the clinical measure of instantaneous wave-free flow ratio (iFR), was investigated. Results have shown that post-dilations are necessary for the lumen gain as well as the hemodynamic restoration towards hemostasis. Malapposed struts induced much higher shear rate, flow disturbances and lower time-averaged wall shear stress (TAWSS) around struts. Post-dilations mitigated the strut malapposition, and thus the shear rate. Moreover, stenting induced larger area of low TAWSS (<0.4 Pa) and lager volume of high shear rate (>2000 s-1), indicating higher risks of in-stent restenosis (ISR) and stent thrombosis (ST), respectively. Oscillatory shear index (OSI) and relative residence time (RRT) indicated the wall regions more prone to ISR are located near the malapposed stent struts.


Clinical outcomes of Modified Collaborative Ocular Melanoma Study IRIS plaques for treatment of iris, iridociliary, and ciliary body melanoma.
To report clinical outcomes of modified Collaborative Ocular Melanoma Study IRIS (COMS IRIS) plaques for treatment of iris, iridociliary, and ciliary body melanoma. Retrospective, single-centre cohort study of iris melanoma treated with COMS IRIS plaque radiotherapy from July 26, 2010 to October 15, 2018. Medical records were reviewed for demographics, tumour features, treatment parameters, and clinical outcomes. There were 22 cases, diagnosed at mean age of 59 years (median 65, range 21-85 years) with female sex in 14 (64%). Presenting tumour features included Snellen visual acuity (VA) ≥ 20/40 in 18 (82%) cases, mean largest tumour basal diameter 4.7 mm (median 3.9, range 2.3-13.8 mm) and thickness 1.7 mm (median 1.6 mm, range 0.8-3.9 mm), iris stromal seeding in 3 (14%) cases, angle seeding in 16 (73%), and ciliary body involvement in 13 (59%). After mean follow-up of 51 months (median 44, range 4-113 months), Snellen VA was ≥20/40 in 14 (64%) cases, with local tumour recurrence in 2 (9%), and enucleation in 2 (9%). The 3-year Kaplan-Meier estimated risk of local tumour recurrence was 7%. The most common radiation side effects were cataract in 17 (77%) patients and dry eye in 5 (23%). Systemic metastasis occurred in no cases, and 1 (5%) non-melanoma-related death due to natural causes was observed at last follow-up. COMS IRIS plaques are effective for treatment of iris, iridociliary, and ciliary body melanoma with modest VA outcomes and low frequency of local tumour recurrence, enucleation, radiation side effects, and systemic metastasis.


The effects of a single dose and prolonged benzene administration on 5-hydroxytryptamine and 5-hydroxyindolacetic acid content in rat brain.
The contents of serotonin (5-HT) and its major metabolite, 5-hydroxyindolacetic acid (5-HIAA), in the hypothalamus, the striatum and the remainder of the brain, after a single dose and prolonged administration of benzene were studied in rats. Benzene was injected s.c. in doses of 1/20-1/5 LD50 for single administration and in doses of 1/80-1/20 LD50 daily six days a week during four weeks of prolonged administration. Single administration produced a decrease in the 5-HT content of the hypothalamus and striatum, accompanied by an increase of the 5-HIAA level in the striatum. The highest contents of both 5-HT and 5-HIAA in the hypothalamus and the striatum were seen 45 min after benzene injection and lasted for at least 180 min. After prolonged benzene administration increases of both 5-HT and 5-HIAA contents were observed, particularly with striatum. The accelerated 5-HT turnover in rat brain after acute and prolonged administration of benzene indicates an effect of the hydrocarbon on the metabolism of 5-HT.


Zinc transport in rice: how to balance optimal plant requirements and human nutrition.
Zinc (Zn) is an essential micronutrient for both plants and animals, while its deficiency in crops and humans is a global problem that affects both crop productivity and human health. Since plants and humans differ in their Zn requirements, it is crucial to balance plant nutrition and human nutrition for Zn. In this review, we focus on the transport system of Zn from soil to grain in rice (Oryza sativa), which is a major dietary source of Zn for people subsiding on rice-based diets. We describe transporters belonging to the different families that are involved in the uptake, vacuolar sequestration, root-to-shoot translocation, and distribution of Zn, and discuss their mechanisms of regulation. We give examples for enhancing Zn accumulation and bioavailability in rice grains through the manipulation of genes that are highly expressed in the nodes, where Zn is deposited at high concentrations. Finally, we provide our perspectives on breeding rice cultivars with both increased tolerance to Zn-deficiency stress and high Zn density in the grains.


Modulation of activation and inactivation by Ca2+ and 2-APB in the pore of an archetypal TRPM channel from Nematostella vectensis.
The archetypal TRPM2-like channel of the sea anemone Nematostella vectensis is gated by ADPR like its human orthologue but additionally exhibits properties of other vertebrate TRPM channels. Thus it can help towards an understanding of gating and regulation of the whole subfamily. To elucidate further the role of Ca2+ as a co-factor of ADPR, we exploited 2-aminoethyl diphenylborinate (2-APB), previously shown to exert either inhibitory or stimulatory effects on diverse TRPM channels, or both in a concentration-dependent manner. 2-APB in high concentrations (1 mM) induced large, non-inactivating currents through nvTRPM2. In lower concentrations (≤0.5 mM), it prevented the fast current inactivation typical for nvTRPM2 stimulated with ADPR. Both these effects were rapidly reversed after wash-out of 2-APB, in contrast to a considerable lag time of their onset. A detailed analysis of nvTRPM2 mutants with modified selectivity filter or reduced ADP-ribose sensitivity revealed that the actions of 2-APB depend on its access to the pore which is enhanced by channel opening. Moreover, access of Ca2+ to the pore is decisive which again depends on the open state of the channel. We conclude that separate regulatory processes by Ca2+ on the pore can be discriminated with the aid of 2-APB.


[Pathogenesis of psoriasis].
Psoriasis is a polymorphous disease. The multiple ways to combine susceptibility genes, immunological mechanisms and modifying factors which interact toward the development of the lesions contribute widely to this polymorphism. It is elusive to look for the source of the disease in an exclusive disorder of the immune system or in an isolated primitive change of the epithelial or stromal skin cells. It is more likely that various combinations of selective abnormalities of these two compartments give raise to the psoriatic phenotype. Indeed, if in on hand T-cells are essential in the development of psoriatic plaques, the role of innate immunity in this process is better recognized, and numerous psoriasis susceptibility genes are linked to immunity, on the other hand some susceptibility factors related to primitive abnormalities of keratinocytes and some of the most recent murine models of psoriasis are based on modifications targeted to the keratinocytes. This article makes a current point on the physiopathology of the disease.


The discovery of AVIL as a bona fide oncogene in glioblastoma.
Our work in rhabdomyosarcoma led us to the discovery of a novel oncogene, Advillin (AVIL) in glioblastoma. Multiple lines of evidence support that AVIL is an Achilles heel of glioblastoma, with its specific targeting potentially an effective treatment approach for the disease. A new signaling axis was also established.


Estimation of relative fitnesses from relative risk data and the predicted future of haemoglobin alleles S and C.
Epidemiological studies of genetic differences in disease susceptibility often estimate the relative risks (RR) of different genotypes. Here I provide an approach to calculate the relative fitnesses of different genotypes based on RR data so that population genetic approaches may be utilized with these data. Using recent RR data on human haemoglobin beta genotypes from Burkina Faso, this approach is used to predict changes in the frequency of the haemoglobin sickle-cell S and C alleles. Overall, it generally appears that allele C will quickly replace the S allele in malarial environments. Explicit population genetic predictions suggest that this replacement may occur within the next 50 generations in Burkina Faso.


Sequential verification of exogenous protein production in OVA gene-targeted chicken bioreactors.
The chicken has potential as an efficient bioreactor system because of its outstanding protein production capacity and low cost. The CRISPR/Cas9-mediated gene-editing system enables production of highly marketable exogenous proteins in transgenic chicken bioreactors. However, because it takes approximately 18 mo to evaluate the recombinant protein productivity of the bioreactor due to the generation interval from G0 founders to G1 egg-laying hens, to verification of the exogenous protein at the early stage is difficult. Here we propose a system for sequential validation of exogenous protein production in chicken bioreactors as in hatching female chicks as well as in egg-laying hens. We generated chicken OVALBUMIN (OVA) EGFP knock-in (KI) chicken (OVA EGFP KI) by CRISPR/Cas9-mediated nonhomologous end joining at the chicken OVA gene locus. Subsequently, the estrogen analog, diethylstilbestrol (DES), was subcutaneously implanted in the abdominal region of 1-wk-old OVA EGFP KI female chicks to artificially increase OVALBUMIN expression. The oviducts of DES-treated OVA EGFP KI female chicks expressed OVA and EGFP at the 3-wk-old stage (10 d after DES treatment). We evaluated the expression of EGFP protein in the oviduct, along with the physical properties of eggs and egg white from OVA EGFP KI hens. The rapid identification and isolation of exogenous protein can be confirmed at a very early stage and high-yield production is possible by targeting the chicken oviduct.


Genetic diversity in an indigenous horse breed: implications for mating strategies and the control of future inbreeding.
The Franches-Montagnes is an indigenous Swiss horse breed, with approximately 2500 foalings per year. The stud book is closed, and no introgression from other horse breeds was conducted since 1998. Since 2006, breeding values for 43 different traits (conformation, performance and coat colour) are estimated with a best linear unbiased prediction (BLUP) multiple trait animal model. In this study, we evaluated the genetic diversity for the breeding population, considering the years from 2003 to 2008. Only horses with at least one progeny during that time span were included. Results were obtained based on pedigree information as well as from molecular markers. A series of software packages were screened to combine best the best linear unbiased prediction (BLUP) methodology with optimal genetic contribution theory. We looked for stallions with highest breeding values and lowest average relationship to the dam population. Breeding with such stallions is expected to lead to a selection gain, while lowering the future increase in inbreeding within the breed.


Toll like receptors and autoimmunity: a critical appraisal.
There is a constant interplay between the innate and adaptive immune systems, which leads to a protective immune response against pathogens and contributes effectively to self-non-self discrimination. Toll-like receptors (TLRs) are key components of the innate immune system, which activate multiple inflammatory pathways and coordinate systemic defense against pathogens. In addition to recognizing unique molecular patterns associated with different classes of pathogens, TLRs may also recognize a number of self proteins and endogenous nucleic acids. Data originating predominantly from animal models of autoimmune disease and circumstantial data from human patients suggest that inappropriate activation of TLR pathways by endogenous or exogenous ligands may lead to the initiation and/or perpetuation of autoimmune responses and tissue injury.